OA21175A

OA21175A - Inhibitors of APOL1 and methods of using same.

Info

Publication number: OA21175A
Application number: OA1202300064
Authority: OA
Inventors: Jingrong Cao; Jon H. Come; Leslie A. DAKIN; Suganthini S. Nanthakumar; Timothy J. SENTER; Jun Myun AHN; Samantha ANGLE; Micheal Aaron BRODNEY; John E. Cochran; Elena DOLGIKH; Brad D. MAXWELL; Hardwin O'dowd; Jessica Howard OLSEN; Akira Joseph SHIMIZU; Steven David STONE; Haoxuan WANG
Original assignee: Vertex Pharmaceuticals Incorporated
Priority date: 2020-08-26
Filing date: 2021-08-26
Publication date: 2024-02-02

Abstract

The disclosure provides at least one entity chosen from compounds of Formula I, a tautomer thereof, a deuterated derivative of that compound or tautomer, and a pharmaceutically acceptable salt of any of the foregoing, compositions comprising the same, and methods of using the same, including uses in treating APOL1-mediated diseases, including pancreatic cancer, focal segmental glomerulosclerosis (FSGS), and/or non-diabetic kidney disease (NDKD).

Description

INHIBITORS OF APOL1 AND METHODS OF USING SAME

This application daims the benefit of priority of U.S. Provisional Patent Application No. 63/070,705, fïled August 26, 2020, the contents of which are incorporated by reference herein in their entirety.

This disclosure provides compounds that may inhibit apolipoprotein L1 (APOL1) and methods of using those compounds to treat APOLl-mediated diseases, such as, e.g., pancreatic cancer, focal segmentai glomerulosclerosis (FSGS), and/or non-diabetic kidney disease (NDKD). In some embodiments, the FSGS and/or NDKD is associated with at least one of the 2 common APOL1 genetic variants (Gl: S342G:I384M and G2: N388del:Y389del). In some embodiments, the pancreatic cancer is associated with elevated levels of APOL1 (such as, e.g., elevated levels of APOL1 in pancreatic cancer tissues).

FSGS is a rare kidney disease with an estimated global incidence of 0.2 to 1.1/100,000/year. FSGS is a disease of the podocyte (glomerular viscéral épithélial cells) responsible for proteinuria and progressive décliné in kidney fonction. NDKD is a kidney disease involving damage to the podocyte or glomerular vascular bed that is not attributable to diabètes. NDKD is a disease characterized by hypertension and progressive décliné in kidney fonction. Human genetics support a causal rôle for the Gl and G2 APOL1 variants in inducing kidney disease. Individuals with 2 APOL1 alleles are at increased risk of developing end-stage kidney disease (ESKD), including primary (idiopathic) FSGS, human immunodeficiency virus (HlV)-associated FSGS, NDKD, arterionephrosclerosis, lupus nephritis, microalbuminuria, and chronic kidney disease. See, P. Dummer et al., Semin Nephrol. 35(3): 222-236 (2015).

FSGS and NDKD can be divided into different subgroups based on the underlying etiology. One homogeneous subgroup of FSGS is characterized by the presence of independent common sequence variants in the apolipoprotein L1 (APOL1) gene termed Gl and G2, which are referred to as the “APOL1 risk alleles.” Gl encodes a correlated pair of non-synonymous amino acid changes (S342G and I384M), G2 encodes a 2 amino acid délétion (N388del:Y389del) near the C terminus of the protein, and G0 is the ancestral (low risk) allele. A distinct phenotype of NDKD is found in patients with APOL1 genetic risk variants as well. In both APOLl-mediated FSGS and NDKD, higher levels of proteinuria and a more accelerated loss of kidney fonction occur in patients with two risk alleles compared to patients with the same disease who hâve no or just 1 APOL1 genetic risk variant. Altematively in AMKD, higher levels of proteinuria and accelerated loss of kidney fonction can also occur in patients with one risk allele. See, G. Vajgel et al., J. Rheumatol., November 2019, jrheum. 190684.

APOL1 is a 44 kDa protein that is only expressed in humans, gorillas, and baboons. The APOL1 gene is expressed in multiple organs in humans, including the liver and kidney. APOL1 is produced mainly by the liver and contains a signal peptide that allows for sécrétion into the bloodstream, where it circulâtes bound to a subset of high-density lipoproteins. APOL1 is responsible for protection against the invasive parasite, Trypanosoma brucei brucei (T. b. brucei). APOL1 is endocytosed by T. b. brucei and transported to lysosomes, where it inserts into the lysosomal membrane and forms pores that lead to parasite swelling and death.

While the ability to lyse T. b. brucei is shared by ail 3 APOL1 variants (GO, Gl, and G2), APOL1 Gl and G2 variants confer additional protection against parasite species that hâve evolved a sérum résistant associated-protein (SRA) which inhibits APOL1 GO; APOL1 Gl and G2 variants confer additional protection against trypanosoma species that cause sleeping sickness. Gl and G2 variants évadé inhibition by SRA; Gl confers additional protection against T. b. gambiense (which causes West African sleeping sickness) while G2 confers additional protection against T. b. rhodesiense (which causes East African sleeping sickness).

In the kidney, APOL1 is expressed in podocytes, endothélial cells (including glomerular endothélial cells), and some tubular cells. Podocyte-specific expression of APOL1 Gl or G2 (but not GO) in transgenic mice induces structural and fonctional changes, including albuminuria, decreased kidney fonction, podocyte abnormalities, and glomerulosclerosis. Consistent with these data, Gl and G2 variants of APOL1 play a causative rôle in inducing FSGS and accelerating its progression in humans. Individuals with APOL1 risk alleles (i.e., homozygous or compound heterozygous for the APOL1 Gl or APOL1 G2 alleles) hâve increased risk of developing FSGS and they are at risk for rapid décliné in kidney fonction if they develop FSGS. Thus, inhibition of APOL1 could hâve a positive impact in individuals who harbor APOL1 risk alleles.

Although normal plasma concentrations of APOL1 are relatively high and can vary at least 20-fold in humans, circulating APOL1 is not causally associated with kidney disease. However, APOL1 in the kidney is thought to be responsible for the development of kidney diseases, including FSGS and NDKD. Under certain circumstances, APOL1 protein synthesis can be increased by approximately 200-fold by pro-inflammatory cytokines such as interferons or tumor necrosis factor-α. In addition, several studies hâve shown that APOL1 protein can form pH-gated Na⁺/K⁺pores in the cell membrane, resulting in a net efflux of intracellular K⁺, ultimately resulting in activation of local and systemic inflammatory responses, cell swelling, and death.

The risk of ESKD is substantially higher in people of recent sub-Saharan African ancestry as compared to those of European ancestry. In the United States, ESKD is responsible for nearly as many lost years of life in women as from breast cancer and more lost years of life in men than from colorectal cancer.

FSGS and NDKD are caused by damage to podocytes, which are part of the glomerular filtration barrier, resulting in proteinuria. Patients with proteinuria are at a higher risk of developing end-stage kidney disease (ESKD) and developing proteinuria-related complications, such as infections or thromboembolie events. There is no standardized treatment regimen nor approved drugs for FSGS or NDKD. Currently, FSGS and NDKD are managed with symptomatic treatment (including blood pressure control using blockers of the renin angiotensin System), and patients with FSGS and heavy proteinuria may be offered high dose steroids. Current therapeutic options for NDKD are anchored on blood pressure control and blockade of the renin angiotensin System.

Corticosteroids, alone or in combination with other immunosuppressants, induce remission in a minority of patients (e.g., remission of proteinuria in a minority of patients) and are associated with numerous side effects. However, remission is frequently indurable even in patients initially responsive to corticosteroid and/or immunosuppressant treatment. As a resuit, patients, in particular individuals of recent sub-Saharan African ancestry with 2 APOL1 risk alleles, expérience rapid disease progression leading to end-stage rénal disease (ESRD). Thus, there is an unmet medical need for treatment for FSGS and NDKD. Illustratively, in view of evidence that APOL1 plays a causative rôle in inducing and accelerating the progression of kidney disease, inhibition of APOL1 should hâve a positive impact on patients with APOL1 mediated kidney disease, particularly those who carry two APOL1 risk alleles (i.e., are homozygous or compound heterozygous for the G1 or G2 alleles).

Additionally, APOL1 is an aberrantly expressed gene in multiple cancers (Lin et al., Cell Death and Disease (2021), 12:760). Recently, APOL1 was found to be abnormally elevated in human pancreatic cancer tissues compared with adjacent tissues and was associated with poor prognosis in pancreatic cancer patients. In in vivo and in vitro experiments, knockdown of APOL1 significantly inhibited cancer cell prolifération and promoted the apoptosis of pancreatic cancer cells.

One aspect of the disclosure provides at least one compound selected from compounds of Formulae I, lia, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, which can be employed in the treatment of diseases mediated by APOL1, such as

FSGS and NDKD. For example, the at least one compound is a compound represented by Formula I:

(R^3a)k

H wherein X¹, X², R¹, R^3a, R^3b, R⁴, R⁵, k, and m are as defined in an embodiment disclosed herein.

In some embodiments, at least one compound of the disclosure is a compound represented by the following structural formula:

(R^3a)k

R⁴ N R⁵H

Formula I a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, wherein:

X¹ is selected from S and -CR^2a and X² is selected from S and -CR^2b, wherein:

one of X¹ and X² is S;

when X¹ is S, then X² is -CR^2b; and 4 when X² is S, then X¹ is -CR^2a;

R¹ is selected from hydrogen, halogen, -OH, cyano, Ci-Cô alkyl, Ci-Cô alkoxy, C3-C6 cycloalkyl, and phenyl, wherein:

the Ci-Cô alkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, and C1-C4 alkoxy;

the Ci-Cô alkoxy of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen;

the C3-C6 cycloalkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)₂, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂; and the phenyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂; R^2ais selected from hydrogen, halogen, cyano, -OH, =0, and Ci-Cô alkyl, wherein:

the Ci-Cô alkyl of R^2ais optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and C1-C4 alkoxy;

R^2b is selected from hydrogen, halogen, cyano, -OH, =O, and Ci-Cô alkyl;

R^3ais selected from halogen, cyano, -OH, Ci-Cô alkyl, and =O; wherein:

the Ci-Cô alkyl of R^3ais optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

R^3b is selected from C1-C2 alkyl and =0; wherein:

the C1-C2 alkyl of R^3bis optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

------, for each occurrence, is a single bond when R^3ais selected from halogen, cyano, -OH, Ci-Cô alkyl or when R^3b is selected from C1-C2 alkyl; or altematively------, for each occurrence, is a double bond when R^3a is =0 or when R^3b is =0;

R⁴is selected from Ci-Cô alkyl, -C(=O)O(Ci-C4 alkyl), Cz-Cô alkynyl, and (R^a)0-5

, wherein:

the Ci-Cô alkyl of R⁴is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), -C(=O)N(Ci-C₄ alkyl)₂, C₃-C₆ cycloalkyl, 5 to 10-membered heterocyclyl, phenyl, and 5 to 10-membered heteroaryl;

Ring A is selected from C3-C12 carbocyclyl, 3 to 12-membered heterocyclyl, Cô and Cio aryl, and 5 to 10-membered heteroaryl, wherein Ring A is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; wherein:

R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cô alkyl, C2-C6 alkenyl, Ci-Cô alkoxy, Ci-Cô haloalkyl, Ci-Cô haloalkenyl, Ci-C6 haloalkoxy, -C(=O)NR^hR^!, -NR^hR‘, -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NR'Rj, -NR^hS(=O)pR^k-OR^k, -OC(=O)R^k, -OC(=O)OR^k, -OC(=O)NR^hR*, -[O(CH2)q]rO(Ci-C6 alkyl), -S(=O)PR^k, -S(=O)PNR^hRⁱ, -C(=O)OR^k, C3-C12 carbocyclyl, 3- to 12-membered heterocyclyl, Ce and Cio aryl, and 5- to 10-membered heteroaryl; wherein:

the Ci-Cô alkyl, Ci-Cô alkoxy, and the C2-C6 alkenyl of R^a are each optionally substituted with 1 to 3 groups independently selected from Cô to Cio aryl (optionally substituted with 1 to 3 R^m groups), 5- to 10-membered heterocyclyl (optionally substituted with 1 to 3 R^m groups), 5- to 10-membered heteroaryl (optionally substituted with 1 to 3 R^mgroups), cyano, -C(=O)R^k, -C(=O)OR^k, -C(=O)NR^hR^!, -NR^hR', -NR^hC( O)R^k, -NR^hC(=O)OR^k, -NR^1,C(=O)NRⁱRj, -NR^hS(=O)PR^k, -OR^k, -OC(=O)R^k, -OC(=O)OR^k, -OC(=O)NR^hRⁱ, -S(=O)PR^k, -S(=O)_PNR^hR‘, and C3-C6 carbocyclyl (optionally substituted with 1 to 3 R^m groups);

the C3-C12 carbocyclyl, the 3 to 12-membered heterocyclyl, the Cô and Cio aryl, and the 5 to 10-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups independently selected from halogen, cyano, C1-C4 alkyl, -NR^hR*, and -OR^k; wherein:

R^h, R*, and R*, for each occurrence, are each independently selected from hydrogen, C1-C4 alkyl, Cô-Cio aryl, and C3-C6 cycloalkyl; wherein:

the C1-C4 alkyl of any one of R^h, R', and R^J’ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

R^k, for each occurrence, are each independently selected from hydrogen, C1-C4 alkyl, 5- to 10-membered heterocyclyl, and C3-C6 carbocyclyl; wherein:

the C1-C4 alkyl of any one of R^k is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

R^m, for each occurrence, is independently selected from halogen, cyano, oxo, Ci-Cô alkyl, Ci-Cô alkoxy, -S(=O)_PR^k, and -OR^k; wherein:

the Ci-Cô alkyl of R^m is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

R⁵is selected from Ci-Cô alkyl, -C(=O)O(Ci-C4 alkyl), C3-C12 carbocyclyl, 3- to 12membered heterocyclyl, Ce and Cio aryl, and 5- to 10-membered heteroaryl; wherein:

the Ci-Cô alkyl of R⁵is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂;

the C3-C12 carbocyclyl, the 3 to 12-membered heterocyclyl, the Cô and Cio aryl, and the 5 to 10-membered heteroaryl of R⁵ are each optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl) (optionally substituted with -OH), -N(Ci-C4 alkyl)2, C1-C5 alkyl (optionally substituted with -OH), C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), -NHC(=O)(Ci-C₄ alkyl), -C(=O)(Ci-C4 alkoxy), and -C(=O)N(Ci-C4 alkyl)2;

k is an integer selected from 0, 1, and 2, wherein:

when R^3a is selected from halogen, cyano, -OH, and Ci-Cô alkyl, k is 1 or 2; and when R^3a is =O, k is 1 ;

m is an integer selected from 0, 1, and 2, wherein:

when R^3b is selected from C1-C2 alkyl, m is 1 or 2; and when R^3b is =0, m is 1 ;

p is an integer selected from 1 and 2; and q and r are each an integer selected from 1, 2, 3, and 4.

(R^a)o-5

In some embodiments, R⁴ is selected from Ci-Ce alkyl and

In some embodiments, R^sis selected from Ci-Cg alkyl, -C(=O)O(Ci-C4 alkyl), C3-C12 carbocyclyl, 3 to 12-membered heterocyclyl, Ce and Cio aryl, and 5 to 10-membered heteroaryl, wherein:

the Ci-Cô alkyl of R⁵ is optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂;

the C3-C12 carbocyclyl, the 3 to 12-membered heterocyclyl, the Cô and Cio aryl, and the 5- to 10-membered heteroaryl of R⁵ are each optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄alkyl)2.

In some embodiments, at least one compound of the disclosure (e.g., at least one compound of Formula I) is a compound represented by the following structural formula:

Formula lo a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, wherein:

X¹ and X² are each selected from S and -CR², wherein:

one of X¹ and X² is S;

when X¹ is S, then X² is -CR^2b; and when X² is S, then X¹ is -CR^2a;

R¹ is selected from halogen, cyano, Ci-Cô alkyl, Ci-Cô alkoxy, C3-C6 cycloalkyl, and phenyl; wherein:

the Ci-Cô alkyl of R^xis optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, and C1-C4 alkoxy;

the C3-C6 cycloalkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)₂, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂; and the phenyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂; R^2ais selected from hydrogen, halogen, cyano, -OH, =0, and Ci-Cô alkyl; wherein:

R^2bis selected from hydrogen, halogen, cyano, -OH, =0, and Ci-Cô alkyl;

R^3ais selected from halogen, cyano, -OH, Ci-Cô alkyl, and =0; wherein:

R^3b is selected from C1-C2 alkyl and =O; wherein:

------, for each occurrence, is a single bond when R^3ais selected from halogen, cyano, -OH, Ci-Cô alkyl or when R^3b is selected from C1-C2 alkyl; or altematively------, for each occurrence, is a double bond when R^3a is =O or when R^3b is =O;

(R^a)o-5

R⁴ is selected from Ci-Cô alkyl and ---' ; wherein:

the Ci-Cô alkyl of R⁴is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C< alkyl), -C(=O)N(Ci-C< alkyl)₂, C₃-C₆ cycloalkyl, 5 to 10-membered heterocyclyl, phenyl, and 5 to 10-membered heteroaryl;

Ring A is selected from C3-C12 carbocyclyl, 3 to 12-membered heterocyclyl, Ce and Cio aryl, and 5 to 10-membered heteroaryl, wherein Ring A is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; wherein:

R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cô alkyl, C2-C6 alkenyl, Ci-Cô alkoxy, Ci-Cô haloalkyl, Ci-Cô haloalkenyl, Ci-Cô haloalkoxy, -C(=O)NR^hRⁱ, -NR^hR‘, -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NRⁱR^J’, -NR^hS( O)pR^k-OR^k, -OC( O)R^k, -OC(=O)OR^k, -OC(=O)NR^hR‘, -[O(CH2)q]rO(Ci-C₆ alkyl), -S(=O)_PR^k, -S(=O)_PNR^hRⁱ, C3-C12 carbocyclyl, 3 to 12-membered heterocyclyl, Ce and Cio aryl, and 5 to 10membered heteroaryl; wherein:

the Ci-Cô alkyl and the C2-C6 alkenyl of R^a are each optionally substituted with 1 to 3 groups independently selected from cyano, -C(=O)R^k, -C(=O)OR^k, -C(=O)NR^hR‘, -NR^hR‘, -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NRⁱRj, -NR^hS(=O)PR^k, -OR^k, -OC(=O)R^k, -OC(=O)OR^k, -OC(=O)NR^hR‘, -S(=O)PR^k, -S(=O)PNR^hRⁱ, and C3-C₆cycloalkyl;

the C3-C12 carbocyclyl, the 3 to 12-membered heterocyclyl, the

Ce and Cio aryl, and the 5 to 10-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups selected from halogen, cyano, C1-C4 alkyl, -NR^hR‘, and -OR^k; wherein:

R^h, R*, and R^J, for each occurrence, are each independently selected from hydrogen, C1-C4 alkyl, and C3-C6 cycloalkyl; wherein:

the C1-C4 alkyl of any one of R^h, R', and R^j is optionally substituted with 1 to 3 groups selected from halogen, cyano, and -OH;

R^k, for each occurrence, are each independently selected from hydrogen, C1-C4 alkyl, and C3-C6 cycloalkyl; wherein: the C1-C4 alkyl of any one of R^k is optionally substituted with 1 to 3 groups selected from halogen, cyano, and -OH;

R⁵is selected from Ci-Cô alkyl, -C(=O)O(Ci-C4 alkyl), C3-C12 carbocyclyl, 3 to 12membered heterocyclyl, Ce and Cio aryl, and 5 to 10-membered heteroaryl; wherein:

the Ci-Cô alkyl of R⁵ is optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, Ci-C4alkoxy, -C(=O)NH₂, -C(=O)NH(C1-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂;

the C3-C12 carbocyclyl, the 3 to 12-membered heterocyclyl, the Ce and Cio aryl, and the 5 to 10-membered heteroaryl of R⁵ are each optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkyl, Ci-C₄alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄alkyl)2;

k is an integer selected from 0, 1, and 2 when R^3a is selected from halogen, cyano, -OH, Ci-Cô alkyl; or altematively k is an integer selected from 0 and 1 when R^3a is =0;

m is an integer selected from 0, 1, and 2 when R^3b is selected from C1-C2 alkyl; and when R^3b is =0, m is an integer selected from 0 and 1 ;

p is an integer selected from 1 and 2; and q and r are each an integer selected from 1, 2, 3, and 4. In one aspect of the disclosure, the compounds of Formula I are chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), such that the at least one entity is chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), pharmaceutically acceptable salts of any of those compounds, solvatés of any of the foregoing, and deuterated dérivatives of any of the foregoing.

In some embodiments, the disclosure provides pharmaceutical compositions comprising at least one entity chosen from compounds of Formulae I, Ha, Hb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing. In some embodiments, the pharmaceutical compositions may comprise at least one compound chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), pharmaceutically acceptable salts of any of those compounds, solvatés of any of the foregoing, and deuterated dérivatives of any of the foregoing. These compositions may further include at least one additional active pharmaceutical ingrédient and/or at least one carrier.

Another aspect of the disclosure provides methods of treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease) comprising administering to a subject in need thereof, at least one entity chosen from compounds of Formulae I, lia, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, F, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing or a pharmaceutical composition comprising the at least one entity. In some embodiments, the methods comprise administering at least one entity chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing.

Another aspect of the disclosure provides methods of treating an APOLl-mediated cancer (such as, e.g., pancreatic cancer) comprising administering to a subject in need thereof, at least one entity chosen from compounds of Formulae I, Ha, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing or a pharmaceutical composition comprising the at least one entity. In some embodiments, the methods comprise administering at least one entity chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing.

Another aspect of the disclosure provides methods of treating FSGS and/or NDKD comprising administering to a subject in need thereof, at least one entity chosen from compounds of Formulae I, Ha, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing or a pharmaceutical composition comprising the at least one entity. In some embodiments, the methods comprise administering at least one entity chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing.

In some embodiments, the methods of treatment include administration of at least one additional active agent to the subject in need thereof, either in the same pharmaceutical composition as the at least one entity chosen from compounds of Formulae I, lia, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ha’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, or as separate compositions. In some embodiments, the methods comprise administering at least one entity chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing with at least one additional active agent either in the same pharmaceutical composition or in a separate composition.

Also provided are methods of inhibiting APOL1, comprising administering to a subject in need thereof, at least one entity chosen from compounds of Formulae I, Ha, Ilb, Ilia, Illb, IVa, IVb, V, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing or a pharmaceutical composition comprising the at least one entity. In some embodiments, the methods of inhibiting APOL1 comprise administering at least one entity chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing or a pharmaceutical composition comprising the at least one entity.

Brief Description of the Drawings

FIG. 1 depicts an XRPD diffractogram of Compound 181 Phosphate Sait Hydrate at 25 ± 2°C and 40% RH.

FIG. 2 depicts an XRPD diffractogram of Compound 181 Phosphate Sait Hydrate at 25 ± 2°C and 5% RH (black trace) or 90% (gray trace).

FIG. 3 depicts a TGA thermogram of Compound 181 Phosphate Sait Hydrate.

FIG. 4 depicts a DSC curve of Compound 181 Phosphate Sait Hydrate.

FIG. 5 depicts a solid State ¹³C NMR spectrum of Compound 181 Phosphate Sait Hydrate.

FIG. 6 depicts a solid State ¹⁹F NMR spectrum of Compound 181 Phosphate Sait Hydrate at43%RH.

FIG. 7 depicts the effects of relative humidity on solid State ¹⁹F NMR spectrum of Compound 181 Phosphate Sait Hydrate.

FIG. 8 depicts a solid state ³¹P NMR spectrum of Compound 181 Phosphate Sait Hydrate at 43% RH.

FIG. 9 depicts the effects of relative humidity on solid state ³¹P NMR spectrum of Compound 181 Phosphate Sait Hydrate.

FIG. 10 depicts an XRPD diffractogram of Compound 181 Free Form Monohydrate.

FIG. 11 depicts a TGA thermogram of Compound 181 Free Form Monohydrate.

FIG. 12 depicts a DSC curve of Compound 181 Free Form Monohydrate.

FIG. 13 depicts a solid state ^I3C NMR spectrum of Compound 181 Free Form Monohydrate.

FIG. 14 depicts a solid state ¹³C NMR spectrum of dehydrated Compound 181 Free Form Monohydrate.

FIG. 15 depicts a solid state ¹⁹F NMR spectrum of Compound 181 Free Form Monohydrate.

FIG. 16 depicts a solid state ¹⁹F NMR spectrum of dehydrated Compound 181 Free Form Monohydrate.

FIG. 17 depicts an XRPD diffractogram of Compound 181 Phosphate Sait Methanol Solvaté.

FIG. 18 depicts a solid state ¹³C NMR spectrum of Compound 181 Phosphate Sait Methanol Solvaté.

FIG. 19 depicts a solid state ¹⁹F NMR spectrum of Compound 181 Phosphate Sait Methanol Solvaté.

FIG. 20 depicts a solid State ³¹P NMR spectrum of Compound 181 Phosphate Sait Methanol Solvaté.

FIG. 21 depicts an XRPD diffractogram of Compound 181 Phosphate Sait MEK Solvaté.

FIG. 22 depicts a solid State ¹³C NMR spectrum of Compound 181 Phosphate Sait MEK Solvaté.

FIG. 23 depicts a solid State ¹⁹F NMR spectrum of Compound 181 Phosphate Sait MEK Solvaté.

FIG. 24 depicts an XRPD diffractogram of Compound 174 Phosphate Hemihydrate.

FIG. 25 depicts a TGA thermogram of Compound 174 Phosphate Hemihydrate.

FIG. 26 depicts a DSC curve of Compound 174 Phosphate Hemihydrate.

FIG. 27 depicts a solid State ¹³C NMR spectrum of Compound 174 Phosphate Hemihydrate.

FIG. 28 depicts a solid State ¹³C NMR spectrum of dehydrated Compound 174 Phosphate Hemihydrate.

FIG. 29A depicts a solid State ^3IP NMR spectrum of Compound 174 Phosphate Hemihydrate.

FIG. 29B depicts a solid State ^3IP NMR spectrum of dehydrated Compound 174 Phosphate Hemihydrate.

FIG. 30 depicts an XRPD diffractogram of Compound 174 Hemihydrate.

FIG. 31 depicts a TGA thermogram of Compound 174 Hemihydrate.

FIG. 32 depicts a DSC curve of Compound 174 Hemihydrate.

FIG. 33 depicts a solid state ^I3C NMR spectrum of Compound 174 Hemihydrate.

FIG. 34 depicts a solid state ¹³C NMR spectrum of dehydrated Compound 174 Hemihydrate. .

Detailed Description

Définitions

The ternis “selected from” and “chosen from” are used interchangeably herein.

The term “APOL1,” as used herein, means apolipoprotein L1 protein and the terni “APOL1 ” means apolipoprotein L1 gene.

The term “APOL1 mediated disease” refers to a disease or condition associated with aberrant APOL1 (e.g., certain APOL1 genetic variants; elevated levels of APOL1). In some embodiments, an APOL1 mediated disease is an APOL1 mediated kidney disease. In some embodiments, an APOL1 mediated disease is associated with patients having two APOL1 risk alleles, e.g., patients who are homozygous or compound heterozygous for the Gl or G2 alleles. In some embodiments, an APOL1 mediated disease is associated with patients having one APOL1 risk allele.

The terni “APOL1 mediated kidney disease” refers to a disease or condition that impairs kidney fonction and can be attributed to APOL1. In some embodiments, APOL1 mediated kidney disease is associated with patients having two APOL1 risk alleles, e.g., patients who are homozygous or compound heterozygous for the Gl or G2 alleles. In some embodiments, the APOL1 mediated kidney disease is chosen from ESKD, NDKD, FSGS, HIV-associated nephropathy, arterionephrosclerosis, lupus nephritis, microalbuminuria, and chronic kidney disease. In some embodiments, the APOL1 mediated kidney disease is chronic kidney disease or proteinuria.

The terni “FSGS,” as used herein, means focal segmentai glomerulosclerosis, which is a disease of the podocyte (glomerular viscéral épithélial cells) responsible for proteinuria and progressive décliné in kidney fonction, and associated with 2 common APOL1 genetic variants (Gl: S342G:I384M and G2: N388del:Y389del).

The terni “NDKD,” as used herein, means non-diabetic kidney disease, which is characterized by severe hypertension and progressive décliné in kidney fonction, and associated with 2 commonAPOL1 genetic variants (Gl: S342G:I384M and G2: N388del:Y389del).

The ternis “ESKD” and “ESRD” are used interchangeably herein to refer to end stage kidney disease or end stage rénal disease. ESKD/ESRD is the last stage of kidney disease, i.e., kidney failure, and means that the kidneys hâve stopped working well enough for the patient to survive without dialysis or a kidney transplant. In some embodiments, ESKD/ESRD is associated with two APOL1 risk alleles.

The terni “compound,” when referring to a compound of fois disclosure, refers to a collection of molécules having an identical Chemical structure unless otherwise indicated as a collection of stereoisomers (for example, a collection of racemates, a collection of cis/trans stereoisomers, or a collection of (E) and (Z) stereoisomers), except that there may be isotopic variation among the constituent atoms of the molécules. Thus, it will be clear to those of skill in the art that a compound represented by a particular Chemical structure containing indicated deuterium atoms will also contain lesser amounts of isotopologues having hydrogen atoms at one or more of the designated deuterium positions in that structure. The relative amount of such isotopologues in a compound of fois disclosure will dépend upon a number of factors, including the isotopic purity of reagents used to make the compound and the efficiency of incorporation of isotopes in the varions synthesis steps used to préparé the compound. However, as set forth above, the relative amount of such isotopologues in toto will be less than 49.9% of the compound. In other embodiments, the relative amount of such isotopologues in toto will be less than 47.5%, less than 40%, less than 32.5%, less than 25%, less than 17.5%, less than 10%, less than 5%, less than 3%, less than 1%, or less than 0.5% of the compound.

As used herein, “optionally substituted” is interchangeable with the phrase “substituted or unsubstituted.” In general, the terni “substituted,” whether preceded by the term “optionally” or not, refers to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent. Unless otherwise indicated, an “optionally substituted” group may hâve a substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent chosen from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this disclosure are those that resuit in the formation of stable or chemically feasible compounds.

The term “isotopologue” refers to a species in which the Chemical structure differs from a reference compound only in the isotopic composition thereof. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the présent structures except for the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a ¹³C or ¹⁴C, are within the scope of this disclosure.

Unless otherwise indicated, structures depicted herein are also meant to include ail isomeric forms of the structures, e.g., racemic mixtures, cis/trans isomers, géométrie (or conformational) isomers, such as (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Therefore, géométrie and conformational mixtures of the présent compounds are within the scope of the disclosure. Unless otherwise stated, ail tautomeric forms of the compounds of the disclosure are within the scope of the disclosure.

The term “tautomer,” as used herein, refers to one of two or more isomers of compound that exist together in equilibrium, and are readily interchanged by migration of an atom, e.g., a hydrogen atom, or group within the molécule.

“Stereoisomer,” as used herein, refers to enantiomers and diastereomers.

As used herein, “deuterated dérivative” refers to a compound having the same Chemical structure as a reference compound, but with one or more hydrogen atoms replaced by a deuterium atom (“D” or “²H”). It will be recognized that some variation of natural isotopic abundance occurs in a synthesized compound depending on the origin of Chemical materials used in the synthesis. The concentration of naturally abundant stable hydrogen isotopes, notwithstanding this variation, is small and immaterial as compared to the degree of stable isotopic substitution of deuterated dérivatives described herein. Thus, unless otherwise stated, when a reference is made to a “deuterated dérivative” of a compound of the disclosure, at least one hydrogen is replaced with deuterium at well above its natural isotopic abundance (which is typically about 0.015%). In some embodiments, the deuterated dérivatives of the disclosure hâve an isotopic enrichment factor for each deuterium atom, of at least 3500 (52.5% deuterium incorporation at each designated deuterium), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), or at least 6600 (99% deuterium incorporation).

The term “isotopic enrichment factor,” as used herein, means the ratio between the isotopic abundance and the natural abundance of a specified isotope.

The term “alkyl” or “aliphatic,” as used herein, means a straight-chain (z.e., linear or unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated. Unless otherwise specified, alkyl groups contain 1 to 20 alkyl carbon atoms. In some embodiments, alkyl groups contain 1 to 10 aliphatic carbon atoms. In some embodiments, alkyl groups contain 1 to 8 aliphatic carbon atoms. In some embodiments, alkyl groups contain 1 to 6 alkyl carbon atoms, and in some embodiments, alkyl groups contain 1 to 4 alkyl carbon atoms. In other embodiments, alkyl groups contain 1 to 3 alkyl carbon atoms, and in yet other embodiments, alkyl groups contain 1 to 2 alkyl carbon atoms. In some embodiments, alkyl groups are substituted. In some embodiments, alkyl groups are unsubstituted. In some embodiments, alkyl groups are linear or straight-chain or unbranched. In some embodiments, alkyl groups are branched.

The tenus “cycloalkyl,” or “cyclic alkyl,” as used herein, refer to a monocyclic C3-8 hydrocarbon or a spirocyclic, fused, or bridged bicyclic or tricyclic Cs-i4 hydrocarbon that is completely saturated, wherein any individual ring in said bicyclic ring System has 3 to 7 members. In some embodiments, cycloalkyl groups are substituted. In some embodiments, cycloalkyl groups are unsubstituted. In some embodiments, the cycloalkyl is a C3 to Ci2 cycloalkyl. In some embodiments, the cycloalkyl is a C3 to Cs cycloalkyl. In some embodiments, the cycloalkyl is a C3 to Cô cycloalkyl. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentanyl, and cyclohexyl.

The tenus “carbocyclyl” or “cycloaliphatic,” as used herein, encompass the tenus “cycloalkyl” or “cyclic alkyl,” and refer to a monocyclic C3-8 hydrocarbon or a spirocyclic, fused, or bridged bicyclic or tricyclic Cs-i4 hydrocarbon that is completely saturated, or is partially saturated as in it contains one or more units of unsaturation but is not aromatic, wherein any individual ring in said bicyclic ring System has 3 to 7 members. Bicyclic carbocyclyls include combinations of a monocyclic carbocyclic ring fused to a phenyl. In some embodiments, carbocyclyl groups are substituted. In some embodiments, carbocyclyl groups are unsubstituted. In some embodiments, the carbocyclyl is a C3 to C12 carbocyclyl. In some embodiments, the carbocyclyl is a C3 to Cio carbocyclyl. In some embodiments, the carbocyclyl is a C3 to Cs carbocyclyl.

The terms “heteroalkyl,” or “heteroaliphatic,” as used herein, refer to alkyl or aliphatic groups as defmed above, wherein one or two carbon atoms are independently replaced by one or more of oxygen, sulfur, nitrogen, phosphorus, or Silicon.

The term “alkenyl,” as used herein, means a straight-chain (i.e., linear or unbranched), branched, substituted or unsubstituted hydrocarbon chain that contains one or more double bonds. In some embodiments, alkenyl groups are substituted. In some embodiments, alkenyl groups are unsubstituted. In some embodiments, alkenyl groups are straight-chain. In some embodiments, alkenyl groups are branched.

The term “heterocycle,” “heterocyclyl,” “heterocycloaliphatic,” or “heterocyclic,” as used herein, means non-aromatic (z. e., completely saturated or partially saturated as in it contains one or more units of unsaturation but is not aromatic), monocyclic, or spirocyclic, fused, or bridged bicyclic or tricyclic ring Systems in which one or more ring members is an independently chosen heteroatom. Bicyclic heterocyclyls include the following combinations of monocyclic rings: a monocyclic heteroaryl fused to a monocyclic heterocyclyl; a monocyclic heterocyclyl fused to another monocyclic heterocyclyl; a monocyclic heterocyclyl fused to phenyl; a monocyclic heterocyclyl fused to a monocyclic carbocyclyl/cycloalkyl; and a monocyclic heteroaryl fused to a monocyclic carbocyclyl/cycloalkyl.

In some embodiments, the heterocycle comprises a ring atom substituted with one or more oxo groups (such as, e.g., a C=O group, a S=O group, or a SO2 group).

In some embodiments, the “heterocycle,” “heterocyclyl,” “heterocycloaliphatic,” or “heterocyclic” group has 3 to 14 ring members in which one or more ring members is a heteroatom independently chosen from oxygen, sulfur, nitrogen, and phosphorus. In some embodiments, each ring in a bicyclic or tricyclic ring System contains 3 to 7 ring members. In some embodiments, the heterocycle has at least one unsaturated carbon-carbon bond. In some embodiments, the heterocycle has at least one unsaturated carbon-nitrogen bond. In some embodiments, the heterocycle has one heteroatom independently chosen from oxygen, sulfur, nitrogen, and phosphorus. In some embodiments, the heterocycle has one heteroatom that is a nitrogen atom. In some embodiments, the heterocycle has one heteroatom that is an oxygen atom. In some embodiments, the heterocycle has two heteroatoms that are each independently selected from nitrogen and oxygen. In some embodiments, the heterocycle has three heteroatoms that are each independently selected from nitrogen and oxygen. In some embodiments, heterocycles are substituted. In some embodiments, heterocycles are unsubstituted. In some embodiments, the heterocyclyl is a 3- to 12-memberedheterocyclyl. In some embodiments, the heterocyclyl is a 3- to 10-memberedheterocyclyl. In some embodiments, the heterocyclyl is a 3to 8-membered heterocyclyl. In some embodiments, the heterocyclyl is a 5- to 10-membered heterocyclyl. In some embodiments, the heterocyclyl is a 5- to 8-membered heterocyclyl. In some embodiments, the heterocyclyl is a 5- or 6-membered heterocyclyl. Non-limiting examples of monocyclic heterocyclyls include piperidinyl, piperazinyl, tetrahydropyranyl, azetidinyl, tetrahydrothiophenyl 1,1-dioxide, etc.

The terni “heteroatom” means one or more of oxygen, sulfur, nitrogen, phosphorus, or Silicon (including, e.g., any oxidized form of nitrogen, sulfur, phosphorus, or Silicon; the quatemized form of any basic nitrogen or; a substitutable nitrogen of a heterocyclic ring, for example, N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR⁺ (as in N-substituted pyrrolidinyl)).

The term “unsaturated,” as used herein, means that a moiety has one or more units or degrees of unsaturation. Unsaturation is the State in which not ail of the available valence bonds in a compound are satisfied by substituents and thus the compound contains double or triple bonds.

The term “alkoxy” or “thioalkyl,” as used herein, refers to an alkyl group, as previously defined, wherein one carbon of the alkyl group is replaced by an oxygen (“alkoxy”) or sulfur (“thioalkyl”) atom, respectively, provided that the oxygen and sulfur atoms are linked between two carbon atoms. Non-limiting examples of alkoxy groups include methoxy, ethoxy, methylmethoxy, and the like. A “cyclic alkoxy” refers to a monocyclic, spirocyclic, bicyclic, bridged bicyclic, tricyclic, or bridged tricyclic hydrocarbon that contains at least one alkoxy group, but is not aromatic. Non-limiting examples of cyclic alkoxy groups include tetrahydropyranyl, tetrahydrofuranyl, oxetanyl, 8-oxabicyclo[3.2.1]octanyl, and oxepanyl. In some embodiments, “alkoxy” and/or “thioalkyl” groups are substituted. In some embodiments, “alkoxy” and/or “thioalkyl” groups are unsubstituted.

The terms “haloalkyl,” “haloalkenyl,” and “haloalkoxy,” as used herein, refer to a linear or branched alkyl, alkenyl, or alkoxy, respectively, which is substituted with one or more halogen atoms. Non-limiting examples of haloalkyl groups include -CHF2, -CH2F, -CF3, -CF2-, 20 and perhaloalkyls, such as -CF2CF3. Non-limiting examples of haloalkoxy groups include -OCHF2, -OCH₂F, -OCF3, and -OCF₂.

The term “halogen” includes F, Cl, Br, and I, i.e., fluoro, chloro, bromo, and iodo, respectively.

The term “aminoalkyl” means an alkyl group which is substituted with or contains an amino group.

As used herein, an “amino” refers to a group which is a primary, secondary, or tertiary amine.

As used herein, a “carbonyl” group refers to C=O.

As used herein, a “cyano” or “nitrile” group refer to -C=N.

As used herein, a “hydroxy” group refers to -OH.

As used herein, a “thiol” group refers to -SH.

As used herein, “tert” and “t-” each refer to tertiary.

As used herein, “aromatic groups” or “aromatic rings” refer to Chemical groups that contain conjugated, planar ring Systems with delocalized pi électron orbitals comprised of [4n+2] p orbital électrons, wherein n is an integer ranging from 0 to 6. Non-limiting examples of aromatic groups include aryl and heteroaryl groups.

The term “aryl” used alone, or as part of a larger moiety as in “arylalkyl,” “arylalkoxy,” or “aryloxyalkyl,” refers to monocyclic or spirocyclic, fused, or bridged bicyclic or tricyclic ring Systems having a total of five to fourteen ring members, wherein every ring in the System is an aromatic ring containing only carbon atoms and wherein each ring in a bicyclic or tricyclic ring System contains 3 to 7 ring members. Non-limiting examples of aryl groups include phenyl (Cé) and naphthyl (Cio) rings. In some embodiments, aryl groups are substituted. In some embodiments, aryl groups are unsubstituted.

The term “heteroaryl,” used alone or as part of a larger moiety as in “heteroarylalkyl” or “heteroarylalkoxy,” refers to monocyclic or spirocyclic, fused, or bridged bicyclic or tricyclic ring Systems having a total of five to fourteen ring members, wherein at least one ring in the System is aromatic, at least one ring in the System contains one or more heteroatoms, and wherein each ring in a bicyclic or tricyclic ring System contains 3 to 7 ring members. Bicyclic heteroaryls include the following combinations of monocyclic rings: a monocyclic heteroaryl fused to another monocyclic heteroaryl; and a monocyclic heteroaryl fused to a phenyl. In some embodiments, heteroaryl groups are substituted. In some embodiments, heteroaryl groups hâve one or more heteroatoms chosen from nitrogen, oxygen, and sulfur. In some embodiments, heteroaryl groups hâve one heteroatom. In some embodiments, heteroaryl groups hâve two heteroatoms. In some embodiments, heteroaryl groups are monocyclic ring Systems having five ring members. In some embodiments, heteroaryl groups are monocyclic ring Systems having six ring members. In some embodiments, heteroaryl groups are unsubstituted. In some embodiments, the heteroaryl is a 3- to 12-membered heteroaryl. In some embodiments, the heteroaryl is a 3- to 10-membered heteroaryl. In some embodiments, the heteroaryl is a 3- to 8membered heteroaryl. In some embodiments, the heteroaryl is a 5- to 10-membered heteroaryl. In some embodiments, the heteroaryl is a 5- to 8-membered heteroaryl. In some embodiments, the heteroaryl is a 5- or 6-membered heteroaryl. Non-limiting examples of monocyclic heteroaryls are pyridinyl, pyrimidinyl, thiophenyl, thiazolyl, isoxazolyl, etc.

In some embodiments, the heteroaryl comprises a ring atom substituted with one or more oxo groups (such as, e.g., a C=O group, a S=O group, or a SO2 group). Illustratively, a nonlimiting example of a heteroaryl group is a benzo[d]oxazol-2(3H)-one group.

Non-limiting examples of useful protecting groups for nitrogen-containing groups, such as amine groups, include, for example, t-butyl carbamate (Boc), benzyl (Bn), tetrahydropyranyl (THP), 9-fluorenylmethyl carbamate (Fmoc) benzyl carbamate (Cbz), acetamide, trifluoroacetamide, triphenylmethylamine, benzylideneamine, and p-toluenesulfonamide. Methods of adding (a process generally referred to as “protecting”) and removing (process generally referred to as “deprotecting”) such amine protecting groups are well-known in the art and available, for example, in P. J. Kocienski, Protecting Groups, Thieme, 1994, which is hereby incorporated by reference in its entirety and in Greene and Wuts, Protective Groups in Organic Synthesis, 3rd Edition (John Wiley & Sons, New York, 1999) and 4^th Edition (John Wiley & Sons, New Jersey, 2014).

Non-limiting examples of suitable solvents that may be used in methods of this disclosure include, but are not limited to, water, methanol (MeOH), éthanol (EtOH), dichloromethane or “methylene chloride” (CH2CI2), toluene, acetonitrile (MeCN), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), methyl acetate (MeOAc), ethyl acetate (EtOAc), heptane, isopropyl acetate (IPAc), teri-butyl acetate (Z-BuOAc), isopropyl alcohol (IPA), tetrahydrofuran (THF), 2methyl tetrahydrofuran (2-Me THF), methyl ethyl ketone (MEK), terributanol, diethyl ether (Et20), methyl-ieri-butyl ether (MTBE), 1,4-dioxane, and Wmethyl pyrrolidone (NMP).

Non-limiting examples of suitable bases that may be used in methods of this disclosure include, but are not limited to, l,8-diazabicyclo[5.4.0]undec-7-ene (DBU), potassium tertbutoxide (KOtBu), potassium carbonate (K2CO3), Wmethylmorpholine (NMM), triethylamine (EtsN; TEA), diisopropyl-ethyl amine (z-Pr2EtN; DIPEA), pyridine, potassium hydroxide (KOH), sodium hydroxide (NaOH), lithium hydroxide (LiOH) and sodium methoxide (NaOMe; NaOCH₃).

The disclosure includes pharmaceutically acceptable salts of the disclosed compounds. A sait of a compound is formed between an acid and a basic group of the compound, such as an amino functional group, or a base and an acidic group of the compound, such as a carboxyl functional group.

The term “pharmaceutically acceptable,” as used herein, refers to a component that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and other mammals without undue toxicity, irritation, allergie response, and the like, and are commensurate with a reasonable benefit/risk ratio. A “pharmaceutically acceptable sait” means any non-toxic sait that, upon administration to a récipient, is capable of providing, either directly or indirectly, a compound of this disclosure. Suitable pharmaceutically acceptable salts are, for example, those disclosed in S. M. Berge, et al. J. Pharmaceutical Sciences, 1977, 66, 1 to 19.

Acids commonly employed to form pharmaceutically acceptable salts include inorganic acids such as hydrogen bisulfide, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, and phosphoric acid, as well as organic acids such as para-toluenesulfonic acid, salicylic acid, tartaric acid, bitartaric acid, ascorbic acid, maleic acid, besylic acid, fumaric acid, gluconic acid, glucuronic acid, formic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, lactic acid, oxalic acid, para-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid, and acetic acid, as well as related inorganic and organic acids. Such pharmaceutically acceptable salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-l,4-dioate, hexyne-l,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, sulfonate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, βhydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene1-sulfonate, naphthalene-2-sulfonate, mandelate, and other salts. In some embodiments, pharmaceutically acceptable acid addition salts include those formed with minerai acids such as hydrochloric acid and hydrobromic acid, and those formed with organic acids such as maleic acid.

Pharmaceutically acceptable salts derived from appropriate bases include alkali métal, alkaline earth métal, ammonium, and N⁺(Cm alkyl)4 salts. This disclosure also envisions the quatemization of any basic nitrogen-containing groups of the compounds disclosed herein. Suitable non-limiting examples of alkali and alkaline earth métal salts include sodium, lithium, potassium, calcium, and magnésium. Further non-limiting examples of pharmaceutically acceptable salts include ammonium, quatemary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate. Other suitable, non-limiting examples of pharmaceutically acceptable salts include besylate and glucosamine salts.

The tenus “patient” and “subject” are used interchangeably herein and refer to an animal, including a human.

The ternis “effective dose” and “effective amount” are used interchangeably herein and refer to that amount of compound that produces the desired effect for which it is administered {e.g., improvement in symptoms of FSGS and/or NDKD, lessening the severity of FSGS and/NDKD or a symptom of FSGS and/or NDKD, and/or reducing progression of FSGS and/or NDKD or a symptom of FSGS and/or NDKD). The exact amount of an effective dose will dépend on the purpose of the treatment and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lloyd (1999) The Art, Science and Technology of Pharmaceutical Compounding).

As used herein, the terni “treatment” and its cognâtes refer to slowing or stopping disease progression. “Treatment” and its cognâtes as used herein, include, but are not limited to, the following: complété or partial remission, lower risk of kidney failure (e.g, ESRD), and diseaserelated complications (e.g., edema, susceptibility to infections, or thrombo-embolic events). Improvements in or lessening the severity of any of these symptoms can be readily assessed according to methods and techniques known in the art or subsequently developed.

The terms “about” and “approximately,” when used in connection with doses, amounts, or weight percent of ingrédients of a composition or a dosage form, include the value of a specified dose, amount, or weight percent or a range of the dose, amount, or weight percent that is recognized by one of ordinary skill in the art to provide a pharmacological effect équivalent to that obtained from the specified dose, amount, or weight percent.

The at least one entity chosen from compounds of Formulae I, Ha, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of

Formulae Ιο, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing may be administered once daily, twice daily, or three times daily, for example, for the treatment of FSGS. In some embodiments, the compounds of Formulae I, lia, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o) are chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing. In some embodiments, at least one entity chosen from compounds of Formulae I, Ha, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing is administered once daily. In some embodiments, at least one entity chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing is administered once daily. In some embodiments, at least one entity chosen from compounds of Formulae I, Ha, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing is administered twice daily. In some embodiments, at least one entity chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing is administered twice daily. In some embodiments, at least one entity chosen from compounds of Formulae I, Ha, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, Γ, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”,

IVa’”, IVb’”, Ιο, Hao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Hao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing are administered three times daily. In some embodiments, at least one entity chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing is administered three times daily.

In some embodiments, 2 mg to 1500 mg or 5 mg to 1000 mg of at least one entity chosen from compounds of Formulae I, Ha, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Hao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Hao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing are administered once daily, twice daily, or three times daily. In some embodiments, 2 mg to 1500 mg or 5 mg to 1000 mg of at least one entity chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing is administered once daily, twice daily, or three times daily.

One of ordinary skill in the art would recognize that, when an amount of compound is disclosed, the relevant amount of a pharmaceutically acceptable sait form of the compound is an amount équivalent to the concentration of the free base of the compound. The amounts of the compounds, pharmaceutically acceptable salts, solvatés, and deuterated dérivatives disclosed herein are based upon the free base form of the reference compound. For example, “1000 mg of at least one compound chosen from compounds of Formula I and pharmaceutically acceptable salts thereof’ includes 1000 mg of a compound of Formula I and a concentration of a pharmaceutically acceptable sait of compounds of Formula I équivalent to 1000 mg of compounds of Formula I.

As used herein, the term “ambient conditions” means room température, open air condition, and uncontrolled humidity condition.

As used herein, the ternis “crystalline form” and “Form” interchangeably refer to a crystal structure (or polymorph) having a particular molecular packing arrangement in the crystal lattice. Crystalline forms can be identified and distinguished from each other by one or more characterization techniques including, for example, X-ray powder diffraction (XRPD), single crystal X-ray diffraction, solid State nuclear magnetic résonance (SSNMR), differential scanning calorimetry (DSC), infrared radiation (IR), and/or thermogravimetric analysis (TGA). Accordingly, as used herein, the term “Form A of Compound [X]” or “Compound [X] Form A” refers to a unique crystalline form that can be identified and distinguished from other crystalline forms of Compound I by one or more characterization techniques including, for example, X-ray powder diffraction (XRPD), single crystal X-ray diffraction, SSNMR, differential scanning calorimetry (DSC), infrared radiation (IR), and/or thermogravimetric analysis (TGA).

As used herein, the term “SSNMR” refers to the analytical characterization method of solid State nuclear magnetic résonance. SSNMR spectra can be recorded at ambient conditions or at alternative conditions (e.g., at 275 K) on any magnetically active isotope présent in the sample. The typical examples of active isotopes for small molécule active pharmaceutical ingrédients include *H, ²H, ¹³C, ¹⁹F, ³¹P, ¹⁵N, ¹⁴N, ³⁵C1, ^HB, ⁷Li, ¹⁷O, ²³Na, ⁷⁹Br, and ¹⁹⁵Pt.

As used herein, the term “XRPD” refers to the analytical characterization method of Xray powder diffraction. XRPD patterns can be recorded under ambient conditions in transmission or reflection geometry using a diffractometer.

As used herein, the ternis “X-ray powder diffractogram,” “X-ray powder diffraction pattern,” and “XRPD pattern” interchangeably refer to an experimentally obtained pattern plotting signal positions (on the abscissa) versus signal intensifies on the ordinate). For an amorphous material, an X-ray powder diffractogram may include one or more broad signais; and for a crystalline material, an X-ray powder diffractogram may include one or more signais, each identified by its angular value as measured in degrees 2θ (° 2Θ), depicted on the abscissa of an Xray powder diffractogram, which may be expressed as “a signal at... degrees two-theta,” “a signal at [a] two-theta value(s) of...” and/or “a signal at at least... two-theta value(s) selected from ...”

A “signal” or “peak,” as used herein, refers to a point in the XRPD pattern where the intensity as measured in counts is at a local maximum. One of ordinary skill in the art would recognize that one or more signais (or peaks) in an XRPD pattern may overlap and may, for example, not be apparent to the naked eye. Indeed, one of ordinary skill in the art would recognize that some art-recognized methods are capable of and suitable for determining whether a signal exists in a pattern, such as Rietveld refinement.

As used herein, “a signal at... degrees two-theta,” “a signal at [a] two-theta valuef] of ..and/or “a signal at at least... two-theta value(s) selected from ..refer to X-ray reflection positions as measured and observed in X-ray powder diffraction experiments (° 2Θ).

The repeatability of the angular values is in the range of ± 0.2° 2Θ, i.e., the angular value can be at the recited angular value + 0.2 degrees two-theta, the angular value - 0.2 degrees twotheta, or any value between those two end points (angular value +0.2 degrees two-theta and angular value -0.2 degrees two-theta).

As used herein, the ternis “signal intensities” and “peak intensities” interchangeably refer to relative signal intensities within a given X-ray powder diffractogram. Factors that can affect the relative signal or peak intensities include sample thickness and preferred orientation (e.g., the crystalline particles are not distributed randomly).

As used herein, the term “DSC” refers to the analytical method of Differential Scanning Calorimetry.

As used herein, the term “TGA” refers to the analytical method of Thermo Gravimétrie (or thermogravimetric) Analysis.

As used herein, a “crystalline hydrate” is a crystal form comprising either stoichiometric or nonstoichiometric water in the crystal lattice. In the case of nonstoichiometric hydrate, the amount of water présent in a crystalline hydrate may vary as a function of at least the relative humidity (“RH”). The presence (or absence) of water or different amounts of water may lead to X-ray diffractogram peak position shifts, or the appearance or disappearance of peaks. The presence (or absence) of water or different amount of water may lead to peak shifts or even appearances of new peaks in proton, carbon, fluorine, phosphorus, nitrogen, chlorine (or other NMR active nuclei) solid State NMR spectra.

Compounds and Compositions

In some embodiments, at least one entity of the disclosure is a compound represented by the following structural formula:

one of X¹ and X² is S;

when X¹ is S, then X² is -CR^2b; and when X² is S, then X¹ is -CR^2a;

R¹ is selected from halogen, cyano, Ci-Cô alkyl, Ci-Cô alkoxy, C3-C6 cycloalkyl, and phenyl, wherein:

the Ci-Cô alkyl of RHs optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, and C1-C4 alkoxy;

the C3-C6 cycloalkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)₂, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂; and the phenyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂; R^2a is selected from hydrogen, halogen, cyano, -OH, =0, and Ci-Cô alkyl, wherein:

R^2b is selected from hydrogen, halogen, cyano, -OH, =O, and Ci-Cô alkyl;

R^3ais selected from halogen, cyano, -OH, Ci-Cô alkyl, and =0; wherein:

R^3bis selected from C1-C2 alkyl and =O; wherein:

the C1-C2 alkyl of R^3b is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

-----=, for each occurrence, is a single bond when R^3a is selected from halogen, cyano, -OH, Ci-Cô alkyl or when R^3bis selected from C1-C2 alkyl; or altematively----for each occurrence, is a double bond when R^3a is =0 or when R^3b is =O;

R⁴is selected from Ci-Cô alkyl, -C(=O)O(Ci-C4 alkyl), C2-C6 alkynyl, and

R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cô alkyl, C2-C6 alkenyl, Ci-Cô alkoxy, Ci-Cô haloalkyl, Ci-Cô haloalkenyl, Ci-Cô haloalkoxy, -C(=O)NR^hRⁱ, -NR^hR', -NR^hC(=0)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NR'Rj, -NR^hS( O)pR^k-OR^k, -OC(=O)R^k, -OC(=O)OR^k, -OC(=O)NR^hR‘, -[O(CH2)q]rO(Ci-Cô alkyl), -S(=O)PR^k, -S(=O)PNR^hRⁱ, -C(=O)OR^k, C3-C12 carbocyclyl, 3- to 12-membered heterocyclyl, Cô and Cio aryl, and 5- to 10-membered heteroaryl; wherein:

the Ci-Cô alkyl, Ci-Cô alkoxy, and the C2-C6 alkenyl of R^a are each optionally substituted with 1 to 3 groups independently selected from Cô to Cio aryl (optionally substituted with 1 to 3 R^m groups), 5- to 10-membered heterocyclyl (optionally substituted with 1 to 3 R^m groups), 5- to 10-membered heteroaryl (optionally substituted with 1 to 3 R^mgroups), cyano, -C(=O)R^k, -C(=O)OR^k, -C(=O)NR^hRⁱ, -NR^hR‘, -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NR‘R^J, -NR^hS(=O)PR^k, -OR^k, -OC(=O)R^k, -OC(=O)OR^k, -OC(=O)NR^hR^!, -S(=O)PR^k, -S(=O)_PNR^hR‘, and C3-C6 carbocyclyl (optionally substituted with 1 to 3 R^m groups);

the C3-C12 carbocyclyl, the 3 to 12-membered heterocyclyl, the Ce and Cio aryl, and the 5 to 10-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups independently selected from halogen, cyano, C1-C4 alkyl, -NR^hR*, and -OR^k; wherein:

R^h, R*, and R^j, for each occurrence, are each independently selected from hydrogen, C1-C4 alkyl, Cô-Cio aryl, and C3-C6 cycloalkyl; wherein:

the C1-C4 alkyl of any one of R^h, R', and R* is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

R⁵is selected from Ci-Cô alkyl, -C(=O)O(Ci-C4 alkyl), C3-C12 carbocyclyl, 3- to 12membered heterocyclyl, Cô and Cio aryl, and 5- to 10-membered heteroaryl; wherein:

the Ci-Cô alkyl of R⁵is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4

31, alkoxy,

-C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂;

the C3-C12 carbocyclyl, the 3 to 12-membered heterocyclyl, the Cô and Cio aryl, and the 5 to 10-membered heteroaryl of R⁵ are each optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C₄ alkyl) (optionally substituted with -OH), -N(Ci-C₄ alkyl)2, C1-C5 alkyl (optionally substituted with -OH), Ci-C₄ alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), -NHC(=O)(Ci-C₄ alkyl), -C(=O)(Ci-C₄ alkoxy), and -C(=O)N(Ci-C₄ alkyl)2;

k is an integer selected from 0, 1, and 2, wherein:

when R^3a is selected from halogen, cyano, -OH, and Ci-Cô alkyl, k is 1 or 2; and when R^3a is =0, k is 1 ;

m is an integer selected from 0, 1, and 2, wherein:

In certain embodiments, a compound of the disclosure is represented by one of the following structural formulae:

Formula lia Formula Ilb a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, wherein:

R^2a is selected from hydrogen, halogen, cyano, and Ci-C₄ alkyl; wherein:

the Ci-C₄ alkyl of R^2a is optionally substituted with 1 to 3 groups independently selected from halogen, -OH, and C1-C2 alkoxy;

R^2b is selected from hydrogen, halogen, cyano, and Ci-C₄ alkyl; and k is an integer selected from 0, 1, and 2;

and ail other variables not specifically defined herein are as defined in the foregoing embodiment.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R⁴ is selected from C1-C4 alkyl and (R^a)0-5

---' ; wherein:

the C1-C4 alkyl of R⁴is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C2 alkoxy, C3-C6 cycloalkyl, 5- to 6-membered heterocyclyl, phenyl, and 5 to 6-membered heteroaryl; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R⁴is selected from C1-C2 alkyl and (R^a)o-5

---' ; wherein:

the C1-C2 alkyl of R⁴is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and 5- to 6-membered heterocyclyl;

and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R⁴ is selected from -CH3, -CH2OH, and (tetrahydro-2H-pyran-4-yl)methyl; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

Formula Ilia Formula IHb a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, wherein:

Ring A, for each occurrence, is selected from C3-C6 cycloalkyl, 5- to 10-membered heterocyclyl, phenyl, and 5- to 10-membered heteroaryl; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups;

and ail other variables not specifically defmed herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, Ring A is selected from cyclopropyl, 5- to 10membered heterocyclyl, phenyl, and 5 to 9-membered heteroaryl; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defmed herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, Ring A is selected from cyclopropyl, 5- to 10membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, phenyl, and 5 to 9membered heteroaryl containing 1 to 3 heteroatoms selected from N and O; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, Ring A is selected from cyclopropyl, 5-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 6-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 9-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 10-membered heterocyclyl containing 1 to heteroatoms selected from N and O, phenyl, 5-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O, 6-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O, and 9-membered heteroaryl containing 1 to 3 heteroatoms selected from

N and O; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or

; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups;

In certain embodiments, in a compound, tautomer, deuterated dérivative, or

optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R⁴ is selected from -CH3 and Ring A; wherein

variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R⁵ is selected from C1-C4 alkyl, -C(=O)O(Ci-C2 alkyl), C3-C6 cycloalkyl, and 5 to 10-memberedheterocyclyl; wherein:

the C1-C4 alkyl of R⁵ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and C1-C2 alkoxy; and the Ç3-C6 cycloalkyl and the 5- to 10-membered heterocyclyl of R⁵ are each optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, C1-C2 alkyl, and C1-C2 alkoxy;

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R⁵is selected from C1-C2 alkyl, -C(=O)O(Ci-C2 alkyl), cyclopropyl, cyclobutyl, and 5- to 6-membered heterocyclyl; wherein:

the C1-C2 alkyl of R⁵ is optionally substituted with 1 to 3 groups independently selected from F, Cl, Br, cyano, -OH, and C1-C2 alkoxy; and the cyclopropyl, the cyclobutyl, and the 5 to 6-membered heterocyclyl of R⁵ are each optionally substituted with 1 to 3 groups independently selected from F, Cl, Br, cyano, -OH, C1-C2 alkyl, and C1-C2 alkoxy;

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R⁵ is selected from -CH3, -CH2CH3, -CH2OH, -C(=O)OCH3, -CH2OCH3, -CH(CH₃)2, cyclopropyl, difluorocyclopropyl, and tetrahydro-2H-pyranyl; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, a compound of the disclosure is represented by one of the foliowing structural formulae:

Formula IVa Formula IVb a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing; and ail other variables not specifically defmed herein are as defmed in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R¹ is selected from hydrogen, halogen, cyano, -OH, C1-C4 alkyl, C1-C4 alkoxy, and C3-C6 cycloalkyl; wherein:

the C1-C4 alkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and C1-C2 alkoxy;

the C1-C4 alkoxy of R¹ is optionally substituted with 1 to 3 independently selected halogen groups; and the C3-C6 cycloalkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and C1-C2 alkoxy;

and ail other variables not specifically defmed herein are as defmed in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R¹ is selected from F, Cl, Br, C1-C4 alkyl, and C3-C6 cycloalkyl; wherein:

the C1-C4 alkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen and -OH; and the C3-C6 cycloalkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, and -OH;

the C1-C4 alkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen and -OH; and the C3-C6 cycloalkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen and -OH;

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R¹ is selected from Cl, Br, -CH3, -CF3, -CH2CH3, -CH(CH₃)2, -CH2CHF2, -CH₂CH(CH3)2, difluorocyclobutyl, and cyclohexyl; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R¹ is Cl; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R^3ais selected from halogen, -OH, and C1-C4 alkyl; wherein:

the C1-C4 alkyl of R^3a is optionally substituted with 1 to 3 groups independently selected from halogen and -OH;

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R^3a is selected from F, Cl, Br, -OH, and C1-C2 alkyl; wherein:

the C1-C2 alkyl of R^3a is optionally substituted with 1 to 3 groups independently selected from F, Cl, and -OH;

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R^3a is selected from F, -OH, -CH3, -CHF2, 40 and -CH2OH; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

Formula Va Formula Vb a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cô alkyl, C1-C4 alkoxy, Ci-Cô haloalkyl, Ci-Cô haloalkoxy, -C(=O)NR^hR‘, -NR^hR‘, -NR^hC(=O)R^k, -OR^k, -[O(CH₂)q]rO(Ci-C₆ alkyl), -S(=O)₂R^k, -S(=O)2NR^hR', C3-C6 cycloalkyl, 5 to 10-membered heterocyclyl, phenyl, and 5 to 8-membered heteroaryl; wherein:

the Ci-Cô alkyl of R^a is optionally substituted with 1 to 3 groups independently selected from cyano, -C(=O)NR^hR‘, -NR^hR*, -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NR‘Rj, -NR^hS(=O)PR^k, -OR^k, -S(=O)2R^k, -S(=O)PNR^hRⁱ, and C3-C₆ cycloalkyl;

the Ç3-C6 cycloalkyl, the 5 to 10-membered heterocyclyl, the phenyl, and the 5 to 8membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups independently selected from halogen, C1-C2 alkyl, and -OR^k; wherein:

R^h, R', and R^j, for each occurrence, are each independently selected from hydrogen, C1-C2 alkyl, cyclopropyl, and cyclobutyl; wherein:

the C1-C2 alkyl of any one of R^h, R', and R^J' is optionally substituted with 1 to 3 groups independently selected from halogen and -OH;

R^k, for each occurrence, is each independently selected from hydrogen and C1-C4 alkyl; wherein:

the C1-C4 alkyl of R^k is optionally substituted with 1 to 3 groups independently selected from halogen and -OH; and q and r are each an integer selected from 1,2, and 3; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cô alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, -C(=O)NR^hR‘, -NR^hR‘, -NR^hC(=O)R^k, -OR^k, -[O(CH₂)_q]_rO(Ci-C4 alkyl), -S(=O)₂R^k, -S(=O)2NR^hR*, cyclopropyl, cyclobutyl, 5 to 6-membered heterocyclyl, phenyl, and 5- to 6membered heteroaryl; wherein:

the Ci-Cô alkyl of R^a is optionally substituted with 1 to 3 groups independently selected from cyano, -C(=O)NR^hR', -NR^hR‘, -OR^k, cyclopropyl, and cyclobutyl;

the cyclopropyl, the cyclobutyl, the 5- to 6-membered heterocyclyl, the phenyl, and the 5 to 6-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups independently selected from halogen, -CH3, -OH, and -OCH3; wherein:

R^h and R‘, for each occurrence, are each independently selected from hydrogen, -CH3, cyclopropyl, and cyclobutyl; wherein:

the -CH3 of any one of R^h and R' is optionally substituted with 1 to 3 groups independently selected from F, Cl, and -OH;

R^k, for each occurrence, is each independently selected from hydrogen and

-CH3; wherein:

the -CH3 of R^k is optionally substituted with 1 to 3 groups independently selected from halogen and -OH;

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R^a, for each occurrence, is independently selected from F, Cl, Br, cyano, Ci-Cô alkyl, Ci-C2 alkoxy, Ci-C2 haloalkyl, -C(=O)NR^hR¹, -NR^hR‘, -NR^hC(=O)R^k, -OR^k, -[O(CH2)_q]_rO(Ci-C₂ alkyl), -S(=O)₂R^k, -S(=O)₂NR^hRⁱ, cyclopropyl, cyclobutyl, 5-membered heterocyclyl, phenyl, and 6-membered heteroaryl; wherein:

the Ci-Cô alkyl of R^a is optionally substituted with 1 to 3 groups independently selected from cyano, -C(=O)NR^hRⁱ, -OR^k, and cyclopropyl;

the cyclopropyl, the cyclobutyl, the 5 to 6-membered heterocyclyl, the phenyl, and the 5 to 6-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups independently selected from halogen, -CH3, -OH, and -OCH3; wherein:

R^h and R‘, for each occurrence, are each independently selected from hydrogen, -CH3, and cyclopropyl; wherein:

R^k, for each occurrence, is each independently selected from hydrogen and

-CH₃; and q and r are each an integer selected from 1 and 2; and ail other variables not specifically defined herein are as defmed in any one of the foregoing embodiments.

In certain embodiments, in a compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait of the disclosure, R^a, for each occurrence, is independently selected from F, cyano, -OH, -CH3, -CF3, -CH(CH3)2, -(CH2)2OH, -(CHzhOŒh, -CH₂CH(OH)C₂H₅, -CH2C(CH3)(CH₂OH)2, -OCH3, -OCH2CH3, -[O(CH₂)₂]2OCH3, -CH₂C(=O)NHCH₃, -(CH2)2SO₂CH3, -CH₂C(=O)N(CH3)2, -CH₂(cyclopropyl), -C(=O)NH2, -C(=O)NH(cyclopropyl), -NH2, -NHCH3, -N(CH3)2, -NHC(CH3)2CH2OH, -NHC(=O)CH3, -SO2CH3, -SO2NH2, cyclopropyl, 2-methoxyphenyl, N-methylpiperazinyl, tetrahydro-2H-pyranyl, methylpyrazolyl, pyridinyl, and tetrahydrothiophenyl 1,1-dioxide; and ail other variables not specifically defmed herein are as defined in any one of the foregoing embodiments.

(R^3a)k

Formula I’

a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

Formula Ha”

Formula Ilb”

Formula IVa”

Formula IVb” a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments. ।

acceptable sait of any of the foregoing; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

In certain embodiments, the at least one compound of the disclosure is chosen from Compounds 1 to 220 depicted in Table I, a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing. A wavy line in a compound in Table I (z.e., ) depicts a bond between two atoms and indicates a position of mixed stereochemistry for a collection of molécules, such as a racemic mixture, cis/trans isomers, or (E)/(Z) isomers. An asterisk adjacent to an atom (e.g., ) in a compound in Table I, indicates a chiral position in the molécule.

In certain embodiments, the at least one compound of the disclosure is chosen from Compounds 221 to 391 depicted in Table II, a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing. A wavy line in a compound in Table II (Le., ) depicts a bond between two atoms and indicates a position of mixed stereochemistry for a collection of molécules, such as a racemic mixture, cis/trans isomers, or (E)!(Z) isomers. An asterisk adjacent to an atom (e.g, ) in a compound in Table II, indicates a chiral position in the molécule.

In certain embodiments, the at least one compound of the disclosure is chosen from Compounds 1 to 391 depicted in Table I or II, a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing.

In certain embodiments, the at least one compound of the disclosure is chosen from compounds depicted in Table III, a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing. A wavy line in a compound in Table III (Le., ) depicts a bond between two atoms and indicates a position of mixed stereochemistry for a collection of molécules, such as a racemic mixture, cis/trans isomers, or (E)/(Z) isomers. An asterisk adjacent to an atom (e.g., Table III, indicates a chiral position in the molécule.

) in a compound in i

OH

Ζ'^ζ *4 U ο U \_ Q_^ '. „ 1,0 '. ω / \s ω / \ P \ _π /--\ O—\ CO IZ Λ > Oo T7 Y ΰ -,__/ \Z \ CO \ /\ / en -¹- , X ⁰⁰ IZ x ) )—' \=( \ / \ / o -*· \ /\ / λ γ y ¹ <f -π'Τ'-π „ σ - O n ο _ω 0 — o 1 z'^Z\ _ZÂ z. \ z—Ί -Z.JJ /—Ci ___ r>_

θ^Η \ OH F F ) X V^e' _o ^r f 1 Vv θΧ'⁵ F θχ⁸ F 'h ’v Ό' B 'v -«Ύ B n-n '_n=n ^π 382 383 384 OH ? Χχρ <oq^^ci 'N '·' -Y h [ΓΧ ü Γ^Ρ H _N-N >_n=n H [L 386 387 388 OH QH oOlH^{f c}'Y< H H II ,^N ³⁹⁰ * = 13_C ÇD₃ 391

160

166

165

* Racemic starting material was used, stereochemistry depicted is relative as a mixture of 2 enantiomers ¹ 3:2 mixture 5 ² 4.5:1 mixture ³ 2:1 mixture ⁴ 3:1 mixture ⁵ 5:1 mixture ⁶ 3.5:1 mixture 10 7 2:1 mixture

Some embodiments of the disclosure include dérivatives of Compounds 1 to 391 (e.g., of Compounds 1 to 220) or compounds of Formulae I, lia, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, 15 Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing. In some embodiments, the dérivatives are Silicon dérivatives in which at least one carbon atom in a compound chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220) or compounds of Formulae I, Ha, Ilb, Ilia, IHb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, IHb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, IHb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, IHb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, IHb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, IHb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, has been replaced by Silicon. In some embodiments, the dérivatives are boron dérivatives, in which at least one carbon atom in a compound chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220) or compounds of Formulae I, Ha, Ilb, Ilia, IHb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, IHb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, IHb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, IHb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, IHb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, IHb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, has been replaced by boron. In other embodiments, the dérivatives are phosphorus dérivatives, in which at least one carbon atom in a compound chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220) or compounds of Formulae I, Ha, Ilb, Ilia, IHb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, IHb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, IHb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, IHb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, IHb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, IHb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, has been replaced by phosphorus.

In some embodiments, the dérivative is a Silicon dérivative in which one carbon atom in a compound chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220) or compounds of Formulae I, Ha, Ilb, Ilia, IHb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, IHb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, IHb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, IHb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, IHb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, IHb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, has been replaced by Silicon or a Silicon dérivative (e.g., -Si(CH3)2- or -Si(OH)2-). The carbon replaced by Silicon may be a non-aromatic carbon. In other embodiments, a fluorine has been replaced by Silicon dérivative (e.g., -Si(CH3)3). In some embodiments, the Silicon dérivatives of the disclosure may include one or more hydrogen atoms replaced by deuterium. In some embodiments, a Silicon dérivative of compound chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220) or compounds of Formulae I, Ha, Hb, Ilia, IHb, IVa, IVb, Va, Vb, F, lia’, Ilb’, IHa’, IHb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, IHb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, IHb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g, from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, may hâve Silicon incorporated into a heterocycle ring.

In some embodiments, the dérivative is a boron dérivative in which one carbon atom in a compound chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220) or compounds of Formulae I, Ha, Ilb, Ilia, IHb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, IHb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, IHb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, IHb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, has been replaced by boron or a boron dérivative.

In some embodiments, the dérivative is a phosphorus dérivative in which one carbon atom in a compound chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220) or compounds of Formulae I, Ha, Ilb, Ilia, IHb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, IHb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, IHb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, IHb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, has been replaced by phosphorus or a phosphorus dérivative.

Another aspect of the disclosure provides pharmaceutical compositions comprising at least one compound according to any one formula chosen from Formulae I, Ha, Ilb, Ilia, IHb, IVa, IVb, Va, Vb, Γ, lia’, Ilb’, Ilia’, IHb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, IHb”,

IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o) and Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing. In some embodiments, the pharmaceutical composition comprising at least one compound chosen from Formulae I, Ha, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o) and Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing is administered to a patient in need thereof.

A pharmaceutical composition may farther comprise at least one pharmaceutically acceptable carrier. In some embodiments, the at least one pharmaceutically acceptable carrier is chosen from pharmaceutically acceptable vehicles and pharmaceutically acceptable adjuvants. In some embodiments, the at least one pharmaceutically acceptable is chosen from pharmaceutically acceptable fillers, disintegrants, surfactants, binders, and fabricants.

It will also be appreciated that a pharmaceutical composition of this disclosure can be employed in combination thérapies; that is, the pharmaceutical compositions described herein can further include at least one additional active therapeutic agent. Altematively, a pharmaceutical composition comprising at least one compound chosen from compounds of Formulae I, Ha, Ilb, Ilia, Illb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, Illb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, Illb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, Illb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., from compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing can be administered as a separate composition concurrently with, prior to, or subséquent to, a composition comprising at least one other active therapeutic agent. In some embodiments, a pharmaceutical composition comprising at least one compound chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing can be administered as a separate composition concurrently with, prier to, or subséquent to, a composition comprising at least one other active therapeutic agent.

As described above, pharmaceutical compositions disclosed herein may optionally further comprise at least one pharmaceutically acceptable carrier. The at least one pharmaceutically acceptable carrier may be chosen from adjuvants and vehicles. The at least one pharmaceutically acceptable carrier, as used herein, includes any and ail solvents, diluents, other liquid vehicles, dispersion aids, suspension aids, surface active agents, isotonie agents, thickening agents, emulsifying agents, preservatives, solid binders, and lubricants, as suited to the particular dosage form desired. Remington: The Science and Practice ofPharmacy, 21 st édition, 2005, ed. D.B. Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988 to 1999, Marcel Dekker, New York discloses varions carriers used in formulating pharmaceutical compositions and known techniques for the préparation thereof. Except insofar as any conventional carrier is incompatible with the compounds of this disclosure, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutical composition, its use is contemplated to be within the scope of this disclosure. Non-limiting examples of suitable pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stéarate, lecithin, sérum proteins (such as, e.g., human sérum albumin), buffer substances (such as, e.g., phosphates, glycine, sorbic acid, and potassium sorbate), partial glyceride mixtures of saturated vegetable fatty acids, water, salts, and electrolytes (such as, e.g., protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts), colloïdal silica, magnésium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool fat, sugars (such as, e.g., lactose, glucose, and sucrose), starches (such as, e.g., corn starch and potato starch), cellulose and its dérivatives (such as, e.g., sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate), powdered tragacanth, malt, gelatin, talc, excipients (such as, e.g., cocoa butter and suppository waxes), oils (such as, e.g., peanut oil, cottonseed oïl, safflower oil, sesame oil, olive oil, corn oil, and soybean oil), glycols (such as, e.g., propylene glycol and polyethylene glycol), esters (such as, e.g., ethyl oleate and ethyl laurate), agar, buffering agents (such as, e.g., magnésium hydroxide and aluminum hydroxide), alginic acid, pyrogen-free water, isotonie saline, Ringer's solution, ethyl alcohol, phosphate buffer solutions, non-toxic compatible lubricants (such as, e.g., sodium lauryl sulfate and magnésium stéarate), coloring agents, releasing agents, coating agents, sweetening agents, flavoring agents, perfuming agents, preservatives, and antioxidants.

In some embodiments of the disclosure, the compounds and the pharmaceutical compositions described herein are used to treat FSGS and/or NDKD. In some embodiments, FSGS is mediated by APOL1. In some embodiments, NDKD is mediated by APOL1.

In some embodiments, the methods of the disclosure comprise administering to a patient in need thereof at least one entity chosen from compounds of Formulae I, Ha, Hb, Ilia, IHb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, IHb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, IHb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, IHb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ha’o, Ilb’o, Hla’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Hla’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing. In some embodiments, the compound of Formula I is chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing. In some embodiments, said patient in need thereof possesses APOL1 genetic variants, i.e., Gl: S342G:I384M and G2: N388del:Y389del.

Another aspect of the disclosure provides methods of inhibiting APOL1 activity comprising contacting said APOL1 with at least one entity chosen from compounds of Formulae I, Ha, Ilb, Ilia, IHb, IVa, IVb, Va, Vb, I’, lia’, Ilb’, Ilia’, IHb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, IHb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, IHb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Hla’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o (e.g., compounds of Formulae lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Hla’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing. In some embodiments, the methods of inhibiting APOL1 activity comprise contacting said APOL1 with at least one entity chosen from Compounds 1 to 391 (e.g., from Compounds 1 to 220), a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing.

Non-Limiting Exemplary Embodiments 1

Without limitation, some embodiments of the présent disclosure include:

1. A compound represented by the following structural formula:

one of X¹ and X² is S;

when X¹ is S, then X² is -CR^2b; and when X² is S, then X¹ is -CR^2a;

R¹ is selected from hydrogen, halogen, cyano, -OH, Ci-Cô alkyl, Ci-Ce alkoxy, C3-C6 cycloalkyl, and phenyl, wherein:

the Ci-Ce alkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH₂, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, and C1-C4 alkoxy;

the Ci-Ce alkoxy of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen;

the C3-C6 cycloalkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂; and the phenyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH₂, -NH(Ci-C₄ alkyl), -N(Ci-C₄ alkyl)₂, C1-C4 alkyl, C1-C4 alkoxy, C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂;

R^2ais selected from hydrogen, halogen, cyano, -OH, =O, and Ci-Cô alkyl, wherein:

the Ci-Cô alkyl of R^2a is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and C1-C4 alkoxy;

R^2bis selected from hydrogen, halogen, cyano, -OH, =0, and Ci-Cô alkyl;

R^3a is selected from halogen, cyano, -OH, Ci-Cô alkyl, and =0; wherein:

the Ci-Cô alkyl of R^3a is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

R^3b is selected from C1-C2 alkyl and =0; wherein:

------, for each occurrence, is a single bond when R^3a is selected from halogen, cyano, -OH, Ci-Cô alkyl or when R^3b is selected from C1-C2 alkyl; or altematively------, for each occurrence, is a double bond when R^3a is =0 or when R^3b is =0;

R⁴is selected from Ci-Cô alkyl, -C(=O)O(Ci-C4 alkyl), C2-C6 alkynyl, and (R^a)0-5

, wherein:

the Ci-Cô alkyl of R⁴ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(C1-C₄ alkyl), -C(=O)N(Ci-C₄ alkyl)₂, C₃-C₆ cycloalkyl, 5- to 10membered heterocyclyl, phenyl, and 5- to 10-membered heteroaryl;

Ring A is selected from C3-C12 carbocyclyl, 3- to 12-membered heterocyclyl, Ce and Cio aryl, and 5- to 10-membered heteroaryl, wherein Ring A is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; wherein:

R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cô alkyl, C2-C6 alkenyl, Ci-Cô alkoxy, Ci-Cô haloalkyl, Ci-Cô haloalkenyl, Ci-Cô haloalkoxy, -C(=O)NR^hRⁱ, -NR^hR‘, -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NRⁱRj, -NR^hS(=O)pR^k-OR^k, -OC(=O)R^k, -OC(=O)OR^k, -OC(=O)NR^hR‘, -[O(CH2)q]rO(Ci-C₆ alkyl), -S(=O)_PR^k, -S(=O)_PNR^hRⁱ, -C(=O)OR^k, C3-C12 carbocyclyl, 3- to 12-membered heterocyclyl, Ce and Cio aryl, and 5- to 10membered heteroaryl; wherein:

the Ci-Cô alkyl, Ci-Cô alkoxy, and the Cz-Cô alkenyl of R^a are each optionally substituted with 1 to 3 groups independently selected from Ce to Cio aryl (optionally substituted with 1 to 3 R^m groups), 5- to 10-membered heterocyclyl (optionally substituted with 1 to 3 R^mgroups), 5- to 10-membered heteroaryl (optionally substituted with 1 to 3 R^m groups), cyano, -C(=O)R^k, -C(=O)OR^k, -C(=O)NR^hR‘, -NR^hR‘, -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NRⁱRj, -NR^hS(=O)_PR^k, -OR^k, -OC(=O)R^k, -OC(=O)OR^k, -OC(=O)NR^hRⁱ,

-S(=O)_PR^k, -S(=O)_PNR^hRⁱ, and C3-C6 carbocyclyl (optionally substituted with 1 to 3 R^mgroups);

the C3-C12 carbocyclyl, the 3- to 12-membered heterocyclyl, the Cô and Clo aryl, and the 5- to 10-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups independently selected from halogen, cyano, C1-C4 alkyl, -NR^hR*, and -OR^k; wherein:

R^h, R¹, and R>, for each occurrence, are each independently selected from hydrogen, C1-C4 alkyl, Cô-Cio aryl, and C3-C6 cycloalkyl; wherein:

the C1-C4 alkyl of any one of R^h, R', and Ri is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

the Ci-Cô alkyl of R⁵is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH₂, -NH(Ci-C₄ alkyl), -N(Ci-C₄ alkyl)₂, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂;

the C3-C12 carbocyclyl, the 3 to 12-membered heterocyclyl, the Cô and Cio aryl, and the 5 to 10-membered heteroaryl of R⁵ are each optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl) (optionally substituted with -OH), -N(Ci-C4 alkyl)2, C1-C5 alkyl (optionally substituted with -OH), C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), -NHC(=O)(Ci-C₄ alkyl), -C(=O)(Ci-C₄ alkoxy), and -C(=O)N(Ci-C₄ alkyl)₂;

k is an integer selected from 0, 1, and 2, wherein:

m is an integer selected from 0, 1, and 2, wherein:

when R^3b is selected from C1-C2 alkyl, m is 1 or 2; and when R^3b is =O, m is 1 ;

2. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 1, wherein the compound is represented by one of the following structural formulae:

H H

Formula Ha Formula Hb or is a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, wherein:

R^2ais selected from hydrogen, halogen, cyano, and C1-C4 alkyl; wherein:

the C1-C4 alkyl of R^2a is optionally substituted with 1 to 3 groups independently selected from halogen, -OH, and C1-C2 alkoxy;

R^2bis selected from hydrogen, halogen, cyano, and C1-C4 alkyl; and k is an integer selected from 0, 1, and 2;

and ail other variables not specifically defmed herein are as defmed in Embodiment 1.

3. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 1 or Embodiment 2, wherein R⁴ is selected from C1-C4 alkyl and

the C1-C4 alkyl of R⁴is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, Ci-C2alkoxy, C3-C6 cycloalkyl, 5- to 6-membered heterocyclyl, phenyl, and 5- to 6-membered heteroaryl; and ail other variables not specifically defined herein are as defmed in Embodiment 1 or Embodiment 2.

4. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 3, wherein R⁴ is selected from C1-C2 alkyl and

and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 3.

5. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 4, wherein R⁴is selected from -CH3, -CH2OH, and (tetrahydro-2H-pyran-4-yl)methyl; and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 4.

6. The compound according to any one of Embodiments 1 to 4, wherein the compound is represented by one of the following structural formulae:

(R^3a)k (R^3a)k

Formula Ilia Formula Illb a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, wherein:

and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 5.

7. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 4 and 6, wherein Ring A is selected from cyclopropyl, 5- to 10-membered heterocyclyl, phenyl, and 5- to 9-membered heteroaryl; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 6.

8. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 4, 6, and 7, wherein Ring A is selected from cyclopropyl, 5 to 10-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, phenyl, and 5 to 9-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 7.

9. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 4 and 6 to 8, wherein Ring A is selected from cyclopropyl, 5-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 6-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 9-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 10-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, phenyl, 5-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O, 6-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O, and 9-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 8.

10. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 4 and 6 to 9, wherein Ring A is selected from

; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups;

and ail other variables not specifically defined herein are as defined in any one of Embodiments to 9.

11. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 4 and 6 to 10, wherein Ring A is selected from

; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 4 and 6 to 10.

12. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 4 and 6 to 11, wherein R⁴is selected from -CH3 and

2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 4 and 6 to 11.

13. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 12, wherein R⁵ is selected from C1-C4 alkyl, -C(=O)O(Ci-C2 alkyl), C3-C6 cycloalkyl, and 5 to 10-membered heterocyclyl; wherein:

the C1-C4 alkyl of R⁵ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and C1-C2 alkoxy; and the C3-C6 cycloalkyl and the 5- to 10-membered heterocyclyl of R⁵ are each optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, C1-C2 alkyl, and C1-C2 alkoxy;

and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 12.

14. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 13, wherein R⁵ is selected from C1-C2 alkyl, -C(=O)O(Ci-C2 alkyl), cyclopropyl, cyclobutyl, and 5- to 6-membered heterocyclyl; wherein:

the C1-C2 alkyl of R⁵is optionally substituted with 1 to 3 groups independently selected from F, Cl, Br, cyano, -OH, and C1-C2 alkoxy; and the cyclopropyl, the cyclobutyl, and the 5- to 6-membered heterocyclyl of R⁵ are each optionally substituted with 1 to 3 groups independently selected from F, Cl, Br, cyano, -OH, C1-C2 alkyl, and Ci-C₂ alkoxy;

and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 13.

15. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 14, wherein R⁵ is selected from -CH3, -CH2CH3, -CH2OH, -C(=O)OCH₃, -CH2OCH3, -CH(CH3)₂, cyclopropyl, difluorocyclopropyl, and tetrahydro-2H-pyranyl; and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 14.

16. The compound according to any one of Embodiments 1 to 4 and 6 to 15, wherein the compound is represented by one of the following structural formulae:

Formula IVa

Formula IVb a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing; and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 4 and 6 to 15.

100

17. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 16, wherein R¹ is selected from hydrogen, halogen, cyano, -OH, C1-C4 alkyl, C1-C4 alkoxy, and C3-C6 cycloalkyl; wherein:

the C1-C4 alkoxy of R¹ is optionally substituted with 1 to 3 independently selected from halogen groups; and the C3-C6 cycloalkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and C1-C2 alkoxy;

and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 16.

18. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 17, wherein R¹ is selected from F, Cl, Br, C1-C4 alkyl, and C3-C6 cycloalkyl; wherein:

and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 17.

19. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 18, wherein R¹ is selected from F, Cl, Br, C1-C4 alkyl, and C3-C6 cycloalkyl; wherein:

the C1-C4 alkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen and -OH;

and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 18.

20. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 18, wherein R¹ is selected from Cl, Br, -CH3, -CF3, -CH2CH3, -CH(CH₃)2, -CH2CHF2, -CH₂CH(CH3)2, difluorocyclobutyl, and cyclohexyl.

101

21. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 20, wherein R¹ is Cl; and ail other variables not specifically defmed herein are as defmed in any one of Embodiments 1 to 20.

22. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 21, wherein R^3ais selected from halogen, -OH, and C1-C4 alkyl; wherein:

the C1-C4 alkyl of R^3ais optionally substituted with 1 to 3 groups independently selected from halogen and -OH;

and ail other variables not specifically defmed herein are as defmed in any one of Embodiments 1 to21.

23. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 22, wherein R^3a is selected from F, Cl, Br, -OH, and C1-C2 alkyl; wherein:

the C1-C2 alkyl of R^3ais optionally substituted with 1 to 3 groups independently selected from F, Cl, and -OH;

and ail other variables not specifically defmed herein are as defmed in any one of Embodiments 1 to 22.

24. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 23, wherein R^3a is selected from F, -OH, -CH3, -CHF2, and CH2OH;and ail other variables not specifically defmed herein are as defmed in any one of Embodiments 1 to 23.

25. The compound according to any one of Embodiments 1 to 4 and 6 to 24, wherein the compound is represented by one of the following structural formulae:

102

Formula Va Formula Vb a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing; and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 4 and 6 to 24.

26. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 4 and 6 to 25, wherein R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cô alkyl, C1-C4 alkoxy, Ci-Cô haloalkyl, Ci-Cô haloalkoxy, -C(=O)NR^hRⁱ, -NR^hRⁱ, -NR^hC(=O)R^k, -OR^k, -[O(CH2)q]rO(Ci-C₆ alkyl), -S(=O)2R^k, -S(=O)2NR^hR‘, C3-C6 cycloalkyl, 5 to 10-membered heterocyclyl, phenyl, and 5- to 8-membered heteroaryl; wherein:

the Ci-Cô alkyl of R^a is optionally substituted with 1 to 3 groups independently selected from cyano, -C(=O)NR^hR^!, -NR^hR‘, -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NRⁱRj, -NR^hS(=O)PR^k -OR^k, -S(=O)2R^k, -S(=O)PNR^hRⁱ, and C3-C₆ cycloalkyl;

the C3-C6 cycloalkyl, the 5 to 10-membered heterocyclyl, the phenyl, and the 5- to 8membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups independently selected from halogen, C1-C2 alkyl, and -OR^k; wherein:

R^h, R*, and R*, for each occurrence, are each independently selected from hydrogen, C1-C2 alkyl, cyclopropyl, and cyclobutyl; wherein:

the C1-C2 alkyl of any one of R^h, R', and R* is optionally substituted with 1 to 3 groups independently selected from halogen and -OH;

the C1-C4 alkyl of R^k is optionally substituted with 1 to 3 groups independently selected from halogen and -OH; and q and r are each an integer selected from 1, 2, and 3;

103 and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 4 and 6 to 25.

27. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 4 and 6 to 26, wherein R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cô alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, -C(=O)NR^hRⁱ, -NR^hR‘, -NR^hC(=O)R^k, -OR^k, -[O(CH2)q]rO(Ci-C4 alkyl), -S(=O)2R^k, -S(=O)2NR^hR¹, cyclopropyl, cyclobutyl, 5- to 6-membered heterocyclyl, phenyl, and 5- to 6-membered heteroaryl; wherein:

the Ci-Cô alkyl of R^a is optionally substituted with 1 to 3 groups independently selected from cyano, -C(=O)NR^hR‘, -NR^hR', -OR^k, cyclopropyl, and cyclobutyl;

R^h and R¹, for each occurrence, are each independently selected from hydrogen,

-CH3, cyclopropyl, and cyclobutyl; wherein:

R^k, for each occurrence, is each independently selected from hydrogen and -CH3; wherein:

and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 4 and 6 to 26.

28. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 4 and 6 to 27, wherein R^a, for each occurrence, is independently selected from F, Cl, Br, cyano, Ci-Cô alkyl, C1-C2 alkoxy, C1-C2 haloalkyl, -C(=O)NR^hRⁱ, -NR^hR‘, -NR^hC(=O)R^k, -OR^k, -[O(CH₂)_q]rO(Ci-C2 alkyl), -S(=O)₂R^k, -S(=O)2NR^hR', cyclopropyl, cyclobutyl, 5-membered heterocyclyl, phenyl, and 6-membered heteroaryl; wherein:

the Ci-Cô alkyl of R^a is optionally substituted with 1 to 3 groups independently selected from cyano, -C(=O)NR^hR‘, -OR^k, and cyclopropyl;

104 the cyclopropyl, the cyclobutyl, the 5- to 6-membered heterocyclyl, the phenyl, and the 5- to 6-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups independently selected from halogen, -CH3, -OH, and -OCH3; wherein:

R^h and R', for each occurrence, are each independently selected from hydrogen, -CH3, and cyclopropyl; wherein:

R^k, for each occurrence, is each independently selected from hydrogen and -CH3; and q and r are each an integer selected from 1 and 2;

and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 4 and 6 to 27.

29. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 4 and 6 to 28, wherein R^a, for each occurrence, is independently selected from F, cyano, -OH, -CH3, -CF₃, -CH(CH₃)2, -(CH₂)₂OH, -(CH₂)₂OCH3, -CH2CH(OH)C₂H₅, -CH₂C(CH3)(CH2OH)2, -OCH3, -OCH2CH3, -[O(CH₂)2]₂OCH3, -CH₂C(=O)NHCH₃, -(CH₂)₂SO₂CH3, -CH₂C(=O)N(CH₃)2, -CH₂(cyclopropyl), -C(=O)NH₂, -C(=O)NH(cyclopropyl),-NH₂, -NHCH3, -N(CH₃)₂, -NHC(CH₃)₂CH₂OH, -NHC(=O)CH₃, -SO2CH3, -SO2NH2, cyclopropyl, 2-methoxyphenyl, N-methylpiperazinyl, tetrahydro-2H-pyranyl, methylpyrazolyl, pyridinyl, and tetrahydrothiophenyl 1,1-dioxide; and ail other variables not specifically defined herein are as defined in any one of Embodiments 1 to 4 and 6 to 28.

30. The compound according to Embodiment 1, wherein the compound is represented by one of the following structural formulae:

(R^3a)k

Formula I’

105

106

31. A compound selected from the compounds of Table I, tautomers thereof, deuterated dérivative of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.

32. A compound selected from the compounds of Table II, tautomers thereof, deuterated dérivative of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.

33. A compound selected from the compounds of Table III, tautomers thereof, deuterated dérivative of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.

34. A pharmaceutical composition comprising at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33 and a pharmaceutically acceptable carrier.

35. A method of treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease comprising administering to a patient in need thereof at least one compound according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34.

107

36. Use of at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34 for the manufacture of a médicament for treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

37. At least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34 for use in treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

38. A method of inhibiting APOL1 activity comprising contacting said APOL1 with at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34.

39. Use of at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34 for the manufacture of a médicament for inhibiting APOL1 activity.

40. At least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34 for use in inhibiting APOL1 activity.

41. A method of treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease) comprising administering to a patient in need thereof at least one compound according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34.

42. The method according to Embodiment 41, wherein the APOL1 -mediated disease is cancer.

43. The method according to Embodiment 41 or Embodiment 42, wherein the APOLl-mediated disease is pancreatic cancer.

108

44. Use of at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34 for the manufacture of a médicament for treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease).

45. The use according to Embodiment 44, wherein the APOLl-mediated disease is cancer.

46. The use according to Embodiment 44 or Embodiment 45, wherein the APOLl-mediated disease is pancreatic cancer.

47. At least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34 for use in treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease).

48. The at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait for use or the pharmaceutical composition for use according to Embodiment 47, wherein the APOLl-mediated disease is cancer.

49. The at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait for use or the pharmaceutical composition for use according to Embodiment 47 or Embodiment 48, wherein the APOLl-mediated disease is pancreatic cancer.

50. A method of inhibiting APOL1 activity comprising contacting said APOL1 with at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34.

51. Use of at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34 for the manufacture of a médicament for inhibiting APOL1 activity.

52. At least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33 or a pharmaceutical composition according to Embodiment 34 for use in inhibiting APOL1 activity.

109

53. A Silicon dérivative of the at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33.

54. A pharmaceutical composition comprising a Silicon dérivative of Embodiment 53.

55. A method of treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease comprising administering to a patient in need thereof a Silicon dérivative according to Embodiment 53 or a pharmaceutical composition according to Embodiment 54.

56. Use of the Silicon dérivative according to Embodiment 53 or a pharmaceutical composition according to Embodiment 54 for the manufacture of a médicament for treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

57. The Silicon dérivative according to Embodiment 53 or a pharmaceutical composition according to Embodiment 54 for use in treating focal segmentai glomerulosclerosis and/or nondiabetic kidney disease.

58. A method of treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease) comprising administering to a patient in need thereof a Silicon dérivative according to Embodiment 53 or a pharmaceutical composition according to Embodiment 54.

59. The method according to Embodiment 58, wherein the APOLl-mediated disease is cancer.

60. The method according to Embodiment 58 or Embodiment 59, wherein the APOLl-mediated disease is pancreatic cancer.

61. Use of the Silicon dérivative according to Embodiment 53 or a pharmaceutical composition according to Embodiment 54 for the manufacture of a médicament for treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease).

62. The use according to Embodiment 61, wherein the APOLl-mediated disease is cancer.

110

63. The use according to Embodiment 61 or Embodiment 62, wherein the APOLl-mediated disease is pancreatic cancer.

64. The Silicon dérivative according to Embodiment 53 or a pharmaceutical composition according to Embodiment 54 for use in treating an APOLl-mediated disease (e.g., an APOLlmediated kidney disease).

65. The Silicon dérivative or pharmaceutical composition for use according to Embodiment 64, wherein the APOLl-mediated disease is cancer.

66. The Silicon dérivative or pharmaceutical composition for use according to Embodiment 64 or Embodiment 65, wherein the APOLl-mediated disease is pancreatic cancer.

67. A boron dérivative of the at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33.

68. A pharmaceutical composition comprising a boron dérivative of Embodiment 67.

69. A method of treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease comprising administering to a patient in need thereof a boron dérivative according to Embodiment 67 or a pharmaceutical composition according to Embodiment 68.

70. Use of the boron dérivative according to Embodiment 67 or a pharmaceutical composition according to Embodiment 68 for the manufacture of a médicament for treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

71. The boron dérivative according to Embodiment 67 or a pharmaceutical composition according to Embodiment 68 for use in treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

72. A method of treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease) comprising administering to a patient in need thereof a boron dérivative according to Embodiment 67 or a pharmaceutical composition according to Embodiment 68.

111

73. The method according to Embodiment 72, wherein the APOL1-mediated disease is cancer.

74. The method according to Embodiment 72 or Embodiment 73, wherein the APOL1-mediated disease is pancreatic cancer.

75. Use of the boron dérivative according to Embodiment 67 or a pharmaceutical composition according to Embodiment 68 for the manufacture of a médicament for treating an APOL1-mediated disease (e.g., an APOL1-mediated kidney disease).

76. The use according to Embodiment 75, wherein the APOL1-mediated disease is cancer.

77. The use according to Embodiment 75 or Embodiment 76, wherein the APOL1-mediated disease is pancreatic cancer.

78. The boron dérivative according to Embodiment 67 or a pharmaceutical composition according to Embodiment 68 for use in treating an APOL1-mediated disease (e.g., an APOL1mediated kidney disease).

79. The boron dérivative or pharmaceutical composition for use according to Embodiment 78, wherein the APOL1 -mediated disease is cancer.

80. The boron dérivative or pharmaceutical composition for use according to Embodiment 78 or Embodiment 79, wherein the APOL1-mediated disease is pancreatic cancer.

81. A phosphorus dérivative of at least one compound, tautomer, deuterated dérivative or pharmaceutically acceptable sait according to any one of Embodiments 1 to 33.

82. A pharmaceutical composition comprising a phosphorus dérivative of Embodiment 81.

83. A method of treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease comprising administering to a patient in need thereof a phosphorus dérivative according to Embodiment 81 or a pharmaceutical composition according to Embodiment 82.

112

84. Use of the phosphorus dérivative according to Embodiment 81 or a pharmaceutical composition according to Embodiment 82 for the manufacture of a médicament for treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

85. The phosphorus dérivative according to Embodiment 81 or a pharmaceutical composition according to Embodiment 82 for use in treating focal segmentai glomerulosclerosis and/or nondiabetic kidney disease.

86. A method of treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease) comprising administering to a patient in need thereof a phosphorus dérivative according to Embodiment 81 or a pharmaceutical composition according to Embodiment 82.

87. The method according to Embodiment 86, wherein the APOLl-mediated disease is cancer.

88. The method according to Embodiment 86 or Embodiment 87, wherein the APOLl-mediated disease is pancreatic cancer.

89. Use of the phosphorus dérivative according to Embodiment 81 or a pharmaceutical composition according to Embodiment 82 for the manufacture of a médicament for treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease).

90. The use according to Embodiment 89, wherein the APOLl-mediated disease is cancer.

91. The use according to Embodiment 89 or Embodiment 90, wherein the APOL1 -mediated disease is pancreatic cancer.

92. The phosphorus dérivative according to Embodiment 81 or a pharmaceutical composition according to Embodiment 82 for use in treating an APOLl-mediated disease (e.g., an APOLlmediated kidney disease).

93. The phosphorus dérivative or pharmaceutical composition for use according to Embodiment 92, wherein the APOLl-mediated disease is cancer.

113

94. The phosphorus dérivative or pharmaceutical composition for use according to Embodiment 92 or Embodiment 93, wherein the APOL1-mediated disease is pancreatic cancer.

95. A method of treating a patient; an entity (e.g., a compound, tautomer, deuterated dérivative, pharmaceutically acceptable sait, Silicon dérivative, boron dérivative, phosphorus dérivative) or pharmaceutical composition for use in treating a patient; or use of an entity or pharmaceutical composition in treating a patient as described in any embodiment herein, wherein the patient has 2 APOL1 risk alleles.

96. A method of treating a patient; an entity (e.g., a compound, tautomer, deuterated dérivative, pharmaceutically acceptable sait, Silicon dérivative, boron dérivative, phosphorus dérivative) or pharmaceutical composition for use in treating a patient; or use of an entity or pharmaceutical composition in treating a patient as described in any embodiment herein, wherein the patient has 1 APOL1 risk allele.

97. The compound according to Embodiment 1, wherein the compound is represented by one of the following structural formulae:

Formula lia” Formula Ilb”

114

Formula IVa” Formula IVb” a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

115

98. The compound according to Embodiment 1, wherein the compound is represented by one of the foliowing structural formulae:

_R3a

116 a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing; and ail other variables not specifically defined herein are as defined in any one of the foregoing embodiments.

99. A pharmaceutical composition comprising at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 and a pharmaceutically acceptable carrier.

100. A method of treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease comprising administering to a patient in need thereof at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99.

101. Use of at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99 for the manufacture of a médicament for treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

102. At least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99 for use in treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

103. A method of inhibiting APOL1 activity comprising contacting said APOL1 with at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99.

104. Use of at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99 for the manufacture of a médicament for inhibiting APOL1 activity.

117

105. At least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99 for use in inhibiting APOL1 activity.

106. A method of treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease) comprising administering to a patient in need thereof at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99.

107. The method according to Embodiment 106, wherein the APOLl-mediated disease is cancer.

108. The method according to Embodiment 106 or Embodiment 107, wherein the APOLl-mediated disease is pancreatic cancer.

109. Use of at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99 for the manufacture of a médicament for treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease).

110. The use according to Embodiment 109, wherein the APOLl-mediated disease is cancer.

111. The use according to Embodiment 109 or Embodiment 110, wherein the APOLl-mediated disease is pancreatic cancer.

112. At least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99 for use in treating an APOLl-mediated disease (e.g., an APOLl-mediated kidney disease).

113. The at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait for use or the pharmaceutical composition for use according to Embodiment 112, wherein the APOLl-mediated disease is cancer.

118

114. The at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait for use or the pharmaceutical composition for use according to Embodiment 112 or Embodiment 113, wherein the APOLl-mediated disease is pancreatic cancer.

115. A method of inhibiting APOL1 activity comprising contacting said APOL1 with at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99.

116. Use of at least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99 for the manufacture of a médicament for inhibiting APOL1 activity.

117. At least one compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Embodiment 97 or Embodiment 98 or a pharmaceutical composition according to Embodiment 99 for use in inhibiting APOL1 activity.

Non-Limiting Exemplary Embodiments 2

Without limitation, some embodiments/clauses of the présent disclosure include:

1. A compound represented by the following structural formula:

H

X¹ and X² are each selected from S and -CR², wherein:

119 one of X¹ and X² is S;

when X¹ is S, then X² is -CR^2b; and when X² is S, then X¹ is -CR^2a;

the C3-C6 cycloalkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)₂, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂; and the phenyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH₂, -NH(Ci-C₄ alkyl), -N(Ci-C₄ alkyl)₂, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂; R^2ais selected from hydrogen, halogen, cyano, -OH, =0, and Ci-Cô alkyl; wherein:

R^2bis selected from hydrogen, halogen, cyano, -OH, =0, and Ci-Cô alkyl;

R^3ais selected from halogen, cyano, -OH, Ci-Cô alkyl, and =0; wherein:

R^3b is selected from C1-C2 alkyl and =0; wherein:

------, for each occurrence, is a single bond when R^3a is selected from halogen, cyano, -OH, Ci-Cô alkyl or when R^3b is selected from C1-C2 alkyl; or altematively------, for each occurrence, is a double bond when R^3a is =O or when R^3b is =0;

120 (R^a)o-s

R⁴ is selected from Ci-Cô alkyl and ; wherein:

the Ci-Cô alkyl of R⁴is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), -C(=O)N(Ci-C₄ alkyl)₂, C₃-Cô cycloalkyl, 5 to 10-membered heterocyclyl, phenyl, and 5 to 10-membered heteroaryl;

Ring A is selected from C₃-Ci2 carbocyclyl, 3 to 12-membered heterocyclyl, Cô and Cio aryl, and 5 to 10-membered heteroaryl, wherein Ring A is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; wherein:

R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cô alkyl, C2-C6 alkenyl, Ci-Cô alkoxy, Ci-Cô haloalkyl, Ci-Cô haloalkenyl, Ci-Cô haloalkoxy, -C(=O)NR^hRⁱ, -NR^hR', -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NRⁱRJ, -NR^hS( O)pR^k -OR^k, -OC( O)R^k, -OC(- O)OR^k, -OC(=O)NR^hRⁱ, -[O(CH2)q]rO(Ci-Cô alkyl), -S(=O)PR^k, -S(=O)PNR^hRⁱ, C₃-Ci₂carbocyclyl, 3 to 12-membered heterocyclyl, Cô and Cio aryl, and 5 to 10membered heteroaryl; wherein:

the Ci-Cô alkyl and the C2-C6 alkenyl of R^a are optionally substituted with 1 to 3 groups independently selected from cyano, -C(=O)R^k, -C(=O)OR^k, -C(=O)NR^hRⁱ, -NR*»#, -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NRⁱRj, -NR^hS(=O)PR^k, -OR^k, -OC(=O)R^k, -OC(=O)OR^k, -OC(=O)NR^hR‘, -S(=O)PR^k, -S(=O)PNR^hRⁱ, and C3-Cô cycloalkyl;

the C₃-Ci2 carbocyclyl, the 3 to 12-membered heterocyclyl, the

Cô and Cio aryl, and the 5 to 10-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups selected from halogen, cyano, C1-C4 alkyl, -NR^hR‘, and -OR^k; wherein:

R^h, R*, and R¹, for each occurrence, are each independently selected from hydrogen, C1-C4 alkyl, and C₃-Cô cycloalkyl; wherein:

121 the C1-C4 alkyl of any one of R^h, R*, and R> is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

R^k, for each occurrence, are each independently selected from hydrogen, C1-C4 alkyl, and C3-C6 cycloalkyl; wherein:

the Ci-Cô alkyl of R⁵ is optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, -NH2, -NH(Ci-C₄ alkyl), -N(Ci-C₄ alkyl)₂, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂;

the C3-C12 carbocyclyl, the 3 to 12-membered heterocyclyl, the Ce and Cio aryl, and the 5 to 10-membered heteroaryl of R⁵ are each optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C4 alkyl)2, C1-C4 alkyl, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄alkyl)2;

p is an integer selected from 1 and 2; and q and r are each an integer selected from 1, 2, 3, and 4. _;

2. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Clause 1, wherein the compound is represented by one of the following structural formulae:

122

H H

Formula Ilao Formula Ilbo or is a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, wherein:

R^2ais selected from hydrogen, halogen, cyano, and C1-C4 alkyl; wherein:

the C1-C4 alkyl of R^2a is optionally substituted with 1 to 3 groups selected from halogen, -OH, and C1-C2 alkoxy;

and ail other variables not specifically defmed herein are as defined in Clause 1.

3. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to Clause 1 or Clause 2, wherein R⁴is selected from C1-C4 alkyl and

wherein:

the C1-C4 alkyl of R⁴ is optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, -NH₂, -NH(Ci-C₄ alkyl), -N(Ci-C₄ alkyl)₂, C1-C2 alkoxy, C₃-C₆cycloalkyl, 5 to 6-membered heterocyclyl, phenyl, and 5 to 6-membered heteroaryl;

and ail other variables not specifically defmed herein are as defmed in Clause 1 or Clause 2.

123

4. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 3, wherein R⁴ is selected from C1-C2 alkyl and

the C1-C2 alkyl of R⁴ is optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, and 5 to 6-membered heterocyclyl;

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 3.

5. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 4, wherein R⁴ is selected from -CH3, -CH2OH, and (tetrahydro-2H-pyran-4-yl)methyl; and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 4.

6. The compound according to any one of Clauses 1 to 4, wherein the compound is represented by one of the following structural formulae:

Formula IHao Formula IHbo a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of the foregoing, wherein:

Ring A, for each occurrence, is selected from C3-C6 cycloalkyl, 5 to 10-membered heterocyclyl, phenyl, and 5 to 10-membered heteroaryl; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups;

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 5.

124

7. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 4 and 6, wherein Ring A is selected from cyclopropyl, 5 to 10-membered heterocyclyl, phenyl, and 5 to 9-membered heteroaryl; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 6.

8. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 4, 6, and 7, wherein Ring A is selected from cyclopropyl, 5 to 10-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, phenyl, and 5 to 9-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 7.

9. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 4 and 6 to 8, wherein Ring A is selected from cyclopropyl, 5-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 6-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 9-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 10-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, phenyl, 5-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O, 6-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O, and 9-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 8.

10. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 4 and 6 to 9, wherein Ring A is selected from

125

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 9.

11. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 4 and 6 to 10, wherein Ring A is selected from

126

127 other variables not specifically defined herein are as defined in any one of Clauses 1 to 4 and 6 to 10.

12. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 4 and 6 to 11, wherein R⁴ is selected from -CH3 and Ring wherein Ring A is selected from

N ; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 4 and 6 to 11.

13. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 12, wherein R⁵ is selected from C1-C4 alkyl, -C(=O)O(Ci-C2 alkyl), C3-C6 cycloalkyl, and 5 to 10-membered heterocyclyl; wherein:

the C1-C4 alkyl of R⁵ is optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, and C1-C2 alkoxy; and the C3-C6 cycloalkyl and the 5 to 10-membered heterocyclyl of R⁵ are each optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, C1-C2 alkyl, and C1-C2 alkoxy; and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 12.

14. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 13, wherein R⁵ is selected from C1-C2 alkyl, C(=O)O(Ci-C2 alkyl), cyclopropyl, cyclobutyl, and 5 to 6-membered heterocyclyl; wherein:

the C1-C2 alkyl of R⁵ is optionally substituted with 1 to 3 groups selected from F, Cl, Br, cyano, -OH, and C1-C2 alkoxy; and

128 the cyclopropyl, the cyclobutyl, and the 5 to 6-membered heterocyclyl of R⁵ are each optionally substituted with 1 to 3 groups selected from F, Cl, Br, cyano, -OH, C1-C2 alkyl, and Ci-C2 alkoxy;

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 13.

15. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 14, wherein R⁵is selected from -CH3, -CH2CH3, -CH2OH, -C(=O)OCH₃, -CH2OCH3, -CH(CH₃)2, cyclopropyl, difluorocyclopropyl, and tetrahydro-2H-pyranyl; and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 14.

16. The compound according to any one of Clauses 1 to 4 and 6 to 15, wherein the compound is represented by one of the following structural formulae:

Formula IVao

Formula IVbo a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of the foregoing; and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 4 and 6 to 15.

17. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 16, wherein R¹ is selected from hydrogen, halogen, cyano, -OH, C1-C4 alkyl, C1-C4 alkoxy, and C3-C6 cycloalkyl; wherein:

the C1-C4 alkyl of R¹ is optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, and C1-C2 alkoxy;

129 the C1-C4 alkoxy of R¹ is optionally substituted with 1 to 3 groups of halogen; and the C3-C6 cycloalkyl of R^xis optionally substituted with 1 to 3 groups selected from halogen, cyano, -OH, and C1-C2 alkoxy;

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 16.

18. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 17, wherein R¹ is selected from F, Cl, Br, C1-C4 alkyl, and C3-C6 cycloalkyl; wherein:

the C1-C4 alkyl of R¹ is optionally substituted with 1 to 3 groups selected from halogen and -OH; and the C3-C6 cycloalkyl of R¹ is optionally substituted with 1 to 3 groups selected from halogen, and -OH;

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 17.

19. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 18, wherein R¹ is selected from F, Cl, Br, C1-C4 alkyl, and C3-C6 cycloalkyl; wherein:

the C1-C4 alkyl of R¹ is optionally substituted with 1 to 3 groups selected from halogen and -OH;

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 18.

20. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 18, wherein R¹ is selected from Cl, Br, -CH3, -CF3, -CH2CH3, -CH(CH₃)2, -CH2CHF2, -CH₂CH(CH3)₂, difluorocyclobutyl, and cyclohexyl.

21. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 20, wherein R¹ is Cl; and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 20.

130

22. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 21, wherein R^3ais selected from halogen, -OH, and C1-C4 alkyl; wherein:

the C1-C4 alkyl of R^3a is optionally substituted with 1 to 3 groups selected from halogen and -OH;

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 21.

23. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 22, wherein R^3a is selected from F, Cl, Br, -OH, and C1-C2 alkyl; wherein:

the C1-C2 alkyl of R^3a is optionally substituted with 1 to 3 groups selected from F, Cl, and -OH;

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 22.

24. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 23, wherein R^3ais selected from F, -OH, -CH3, -CHF2, and CH2OH; and ail other variables not specifically defined herein are as defmed in any one of Clauses 1 to 23.

25. The compound according to any one of Clauses 1 to 4 and 6 to 24, wherein the compound is represented by one of the following structural formulae:

Formula Vao

Formula Vbo a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of the foregoing; and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 4 and 6 to 24.

131

26. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 4 and 6 to 25, wherein R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Ce alkyl, C1-C4 alkoxy, Ci-Ce haloalkyl, Ci-Ce haloalkoxy, -C(=O)NR^hRⁱ, -NR^hR', -NR^hC(=O)R^k, -OR^k, -[O(CH₂)q]rO(Ci-C₆ alkyl), -S(=O)2R^k, -S(=O)2NR^hR‘, C3-C6 cycloalkyl, 5 to 10-membered heterocyclyl, phenyl, and 5 to 8membered heteroaryl; wherein:

the Ci-Ce alkyl of R^a is optionally substituted with 1 to 3 groups selected from cyano, -C(=O)NR^hRⁱ, -NR^hR‘, -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -NR^hC(=O)NR'Rj, -NR^hS(=O)PR^k, -OR^k, -S(=O)2R^k, -S(=O)PNR^hR', and C3-C₆ cycloalkyl;

the C3-C6 cycloalkyl, the 5 to 10-membered heterocyclyl, the phenyl, and the 5 to 8membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups selected from halogen, C1-C2 alkyl, and -OR^k; wherein:

R^h, R*, and R^j, for each occurrence, are each independently selected from hydrogen, C1-C2 alkyl, cyclopropyl, and cyclobutyl; wherein:

the C1-C2 alkyl of any one of R^h, R', and R* is optionally substituted with 1 to 3 groups selected from halogen and -OH;

the C1-C4 alkyl of R^k is optionally substituted with 1 to 3 groups selected from halogen and -OH; and q and r are each an integer selected from 1, 2, and 3;

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 4 and 6 to 25.

27. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 4 and 6 to 26, wherein R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Ce alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, -C(=O)NR^hRⁱ, -NR^hR', -NR^hC(=O)R^k, -OR^k, -[O(CH₂)q]rO(Ci-C₄ alkyl), -S(=O)2R^k, -S(=O)2NR^hR‘, cyclopropyl, cyclobutyl, 5 to 6-membered heterocyclyl, phenyl, and 5 to 6-membered heteroaryl; wherein:

the Ci-Ce alkyl of R^a is optionally substituted with 1 to 3 groups selected from cyano, -C(=O)NR^hR', -NR^hR', -OR^k, cyclopropyl, and cyclobutyl;

132 the cyclopropyl, the cyclobutyl, the 5 to 6-membered heterocyclyl, the phenyl, and the 5 to 6-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups selected from halogen, -CH3, -OH, and -OCH3; wherein:

R^h and R*, for each occurrence, are each independently selected from hydrogen, -CH3, cyclopropyl, and cyclobutyl; wherein:

the -CH3 of any one of R^h and R* is optionally substituted with 1 to 3 groups selected from F, Cl, and -OH;

R^k, for each occurrence, is each independently selected from hydrogen and CH3; wherein:

the -CH3 of R^k is optionally substituted with 1 to 3 groups selected from halogen and -OH;

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 4 and 6 to 26.

28. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 4 and 6 to 27, wherein R^a, for each occurrence, is independently selected from F, Cl, Br, cyano, Ci-Cô alkyl, C1-C2 alkoxy, Ci-C2 haloalkyl, -C(=O)NR^hRⁱ, -NRbR¹, -NR^hC(=O)R^k, -OR^k, -[O(CH₂)_q]_rO(Ci-C2 alkyl), -S(=O)₂R^k, -S(=O)2NR^hR‘, cyclopropyl, cyclobutyl, 5-membered heterocyclyl, phenyl, and 6-membered heteroaryl; wherein:

the Ci-Cô alkyl of R^a is optionally substituted with 1 to 3 groups selected from cyano, -C(=O)NR^hR‘, -OR^k, and cyclopropyl;

the cyclopropyl, the cyclobutyl, the 5 to 6-membered heterocyclyl, the phenyl, and the 5 to 6-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups selected from halogen, -CH3, -OH, and -OCH3; wherein:

R^h and R*, for each occurrence, are each independently selected from hydrogen, -CH3, and cyclopropyl; wherein:

the -CH3 of any one of R^h and R' is optionally substituted with 1 to 3 groups selected from F, Cl, and -OH;

and ail other variables not specifically defined herein are as defined in any one of Clauses 1 to 4 and 6 to 27.

133

29. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of Clauses 1 to 4 and 6 to 28, wherein R^a, for each occurrence, is independently selected from F, cyano, -OH, -CH3, -CF3, -CH(CH3)2, -(CH2)2OH, -(CH₂)₂OCH3, -CH₂CH(OH)C2H₅, -CH2C(CH3)(CH₂OH)₂, -OCH3, -OCH2CH3,

-[O(CH₂)2]2OCH₃, -CH₂C(=O)NHCH₃, -(CH₂)2SO₂CH3, -CH₂C(=O)N(CH3)2,

-CH₂(cyclopropyl), -C(=O)NH₂, -C(=O)NH(cyclopropyl), -NH2, -NHCH3, -N(CH₃)₂, -NHC(CH3)₂CH₂OH, -NHC(=0)CH3, -SO2CH3, -SO2NH2, cyclopropyl, 2-methoxyphenyl, N-methylpiperazinyl, tetrahydro-2H-pyranyl, methylpyrazolyl, pyridinyl, and tetrahydrothiophenyl l,l-dioxide;and ail other variables not specifically defined herein are as 10 defined in any one of Clauses 1 to 4 and 6 to 28.

30. The compound according to Clause 1, wherein the compound is represented by one of the following structural formulae:

134

Formula Va’o

Formula Vb’o a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of the foregoing; and ail other variables not specifically defmed herein are as defined in any one of the foregoing Clauses.

135

32. A pharmaceutical composition comprising at least one compound, tautomer, deuterated dérivative or pharmaceutically acceptable sait according to any one of Clauses 1 to 31 and a pharmaceutically acceptable carrier.

33. A method of treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease comprising administering to a patient in need thereof at least one compound according to any one of Clauses 1 to 31 or a pharmaceutical composition according to Clause 32.

34. Use of at least one compound, tautomer, deuterated dérivative or pharmaceutically acceptable sait according to any one of Clauses 1 to 31 or a pharmaceutical composition according to Clause 32 for the manufacture of a médicament for treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

35. At least one compound, tautomer, deuterated dérivative or pharmaceutically acceptable sait according to any one of Clauses 1 to 31 or a pharmaceutical composition according to Clause 32 for use in treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

36. A method of inhibiting APOL1 activity comprising contacting said APOL1 with at least one compound, tautomer, deuterated dérivative or pharmaceutically acceptable sait according to any one of Clauses 1 to 31 or a pharmaceutical composition according to Clause 32.

37. Use of at least one compound, tautomer, deuterated dérivative or pharmaceutically acceptable sait according to any one of Clauses 1 to 31 or a pharmaceutical composition according to Clause 32 for the manufacture of a médicament for inhibiting APOL1 activity.

38. At least one compound, tautomer, deuterated dérivative or pharmaceutically acceptable sait according to any one of Clauses 1 to 31 or a pharmaceutical composition according to Clause 32 for use in inhibiting APOL1 activity.

136

39. A Silicon dérivative of the at least one compound, tautomer, deuterated dérivative or pharmaceutically acceptable sait according to any one of Clauses 1 to 31.

40. A pharmaceutical composition comprising a Silicon dérivative of Clause 39.

41. A method of treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease comprising administering to a patient in need thereof a Silicon dérivative according to Clause 39 or a pharmaceutical composition according to Clause 40.

42. Use of the Silicon dérivative according to Clause 39 or a pharmaceutical composition according to Clause 40 for the manufacture of a médicament for treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

43. The Silicon dérivative according to Clause 39 or a pharmaceutical composition according to Clause 40 for use in treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

44. A boron dérivative of the at least one compound, tautomer, deuterated dérivative or pharmaceutically acceptable sait according to any one of Clauses 1 to 31.

45. A pharmaceutical composition comprising a boron dérivative of Clause 44.

46. A method of treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease comprising administering to a patient in need thereof a boron dérivative according to Clause 44 or a pharmaceutical composition according to Clause 45.

47. Use of the boron dérivative according to Clause 44 or a pharmaceutical composition according to Clause 45 for the manufacture of a médicament for treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

48. The boron dérivative according to Clause 44 or a pharmaceutical composition according to Clause 45 for use in treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

137

49. A phosphorus dérivative of at least one compound, tautomer, deuterated dérivative or pharmaceutically acceptable sait according to any one of Clauses 1 to 31.

50. A pharmaceutical composition comprising a phosphorus dérivative of Clause 48.

51. A method of treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease comprising administering to a patient in need thereof a phosphorus dérivative according to Clause 48 or a pharmaceutical composition according to Clause 49.

52. Use of the phosphorus dérivative according to Clause 48 or a pharmaceutical composition according to Clause 49 for the manufacture of a médicament for treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

53. The phosphorus dérivative according to Clause 48 or a pharmaceutical composition according to Clause 49 for use in treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease.

138

EXAMPLES

In order that the disclosure described herein may be more fùlly understood, the following examples are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this disclosure in any manner.

The compounds of the disclosure may be made according to standard Chemical practices or as described herein. Throughout the following synthetic schemes and in the descriptions for preparing compounds of Formulae I, Ha, Ilb, Ilia, IHb, IVa, IVb, Va, Vb, F, lia’, Ilb’, Ilia’, IHb’, IVa’, IVb’, Va’, Vb’, Ha”, Ilb”, Ilia”, IHb”, IVa”, IVb”, lia’”, Ilb’”, Ilia’”, IHb’”, IVa’”, IVb’”, lo, Ilao, Ilbo, Illao, Illbo, IVao, IVbo, Vao, Vbo, l’o, Ila’o, Ilb’o, Illa’o, Illb’o, IVa’o, IVb’o, Va’o, and Vb’o, Compounds 1 to 391, a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, the following abbreviations are used:

Abbreviations

AIBN = azobisisobutyronitrile

ARP = assay ready plate

BBBPY = 4,4'-Di-Zer/-butyl-2,2'-dipyridyl

BF3 = boron trifluoride

BF3.OEt2 = boro trifluoride diethyl etherate

BOC2O = di-tert-butyl dicarbonate

CBzCl = benzyl chloroformate

CDMT = 2-chloro-4,6-dimethoxy-l,3,5-triazine

DAST = diethylaminosulfur trifluoride

DBU = l,8-diazabicyclo[5.4.0]undec-7-ene

DCM = dichloromethane

DIBAL-H = diisobutylaluminum hydride

DIPEA = Ν,Ν-Diisopropylethylamine or N-ethyl-N-isopropyl-propan-2-amine

DMAP = dimethylamino pyridine

DMA = dimethyl acetamide

DME = dimethoxyethane

DMEM = Dulbecco’s modified Eagle’s medium

DMF = dimethylformamide

DMPU = N,N’-dimethylpropyleneurea

DMSO = dimethyl sulfoxide

139

DPPA = diphenylphosphoryl azide

EtOAc = ethyl acetate

EtOH = éthanol

Et₂O = diethyl ether

FBS = fêtai bovine sérum

FLU = fluorescent values

HATU = [dimethylamino(triazolo [4,5-b]pyridin-3 -yloxy)methylene] -dimethylammonium (Phosphorus Hexafluoride Ion)

HDMC = 7V-[(5-Chloro-3-oxido-177-benzotriazol-l-yl)-4-morpholinylmethylene]- Nmethylmethanaminium hexafluorophosphate

HEPES = 4-(2-hydroxyethyl)-l-piperazineethanesulfonic acid

HBSS = Hank’s balanced sait solution

IPA = isopropyl alcohol

Ir[df(CF3)ppy]₂(dtbbpy)PF6 = phosphorus hexafluoride

LDA = lithium diisopropyl amide

LED = light emitting diode

MeCN = acetonitrile

Mel = methyl iodide

MeOH = methanol

MsOH = methanesulfonic acid

MTBE or TBME = Methyl teri-butyl ether n-BuLi = n-butyllithium

NB S = n-bromosuccinimide

NMM = N-methyl morpholine

NMP = N-methyl pyrrolidine

PBS = phosphate-buffered saline

Pd(dppf)₂Cl₂ = [1,1 '-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)

PdCl₂(PPh3)₂ = Bis(triphenylphosphine)palladium(II) dichloride

PP = polypropylene

PTS A =p-Toluenesulfonic acid monohydrate

140

T3P = 2,4,6-Tripropyl-l,3,5,2,4,6-trioxatriphosphorinane-2,4,6-trioxide

TBAF = tetra-n-butylammonium fluoride

TBSC1 = tert-butyldimethylsilyl chloride

TEA = triethylamine

Tet = tétracycline

TFA or TF AA = trifluoroacetic acid

TfOH = triflic acid

THF = tetrahydrofuran

2-Me-THF = 2=methyltetrahydrofuran

THP = tetrahydropyran

TMSC1 = trimethylsilyl chloride

TMSS = Tris(trimethylsilyl)silane

Example 1. Synthesis of Compounds

Ail the spécifie and generic compounds, and the intermediates disclosed for making those compounds, are considered to be part of the disclosure disclosed herein.

Synthesis of Starting Materials

Préparations describe synthetic routes to intermediates used in the synthesis of Compounds 1 to 391.

General Schemes

In some embodiments, processes for preparing compounds of Formula I comprise the reactions described in Schemes 1-6.

Scheme 1 shows a process for the préparation of compounds of Formula I. R¹, R³, R⁴ R⁵, X¹, X², m, and k are defined as above. An amino ketone of formula 1-1 may undergo reaction with an aldéhyde of formula 1-2 to afford a piperidone of formula 1-3. In some embodiments, the reaction may occur in the presence of an amine catalyst such as L-proline, in the presence of a base such as triethyl amine, and magnésium sulfate reagent. Compounds of formula 1-3 may be prepared using any suitable method for the préparation of a piperidone. A compound of formula 1-3 may be prepared from a piperidone of formula 1-3 and an alcohol of formula 1-4 using any suitable conditions to perform a Pictet-Spengler reaction. For example, the reaction may be performed in the presence of an acid such as trifluoromethyl sulfonic acid and a solvent such as 1,4-dioxane. In an alternative embodiment, an acid such as methanesulfonic acid may be used. The reaction may be performed in a solvent such as dichloromethane in the presence of added

141 heat (e.g., 40 °C).

Scheme 1

Formula I

Scheme 2 depicts processes for the préparation of compounds of formula 2-3. PG¹ is any suitable nitrogen protecting group. For example, in some embodiments, PG¹ is a trifluoroacetate group. A compound of formula 2-2 may be prepared from 2-1 using any suitable method for benzylic oxidation. For example, in some embodiments, the reaction is performed in the presence of oxygen gas under balloon pressure, N-hydroxypthalamide, and cobalt diacetate catalyst. In some embodiments, the reaction is performed in the presence of acetonitrile. The reaction may be performed in the presence of added heat (e.g., at 60 °C). Compounds of formula 2-3 may be prepared from a compound of formula 2-2 using any suitable method for the réduction of a ketone to an alcohol. For example, a Corey-Bakshi-Shibata catalyst (CBS catalyst) in the presence of a reducing agent such as borane may be used. In alternative embodiments, transition métal catalyzed transfer hydrogénation System may be used. In the presence of a chiral ligand, transition métal transfer hydrogénation reaction may resuit in an asymmetric réduction of the ketone.

142

2-1

Scheme 2

2-2

OH

2-3

Scheme 3 shows processes for the préparation of compounds of formula 3-4. PG² is any suitable alcohol protecting group, for example, THP. A heterocyclic bromide of formula 3-1 may 5 be coupled with a trifluoroboronate sait of formula 3-2 using any suitable method for the coupling of a halide with an alkyl boronate. For example, in some embodiments, the reaction may be performed in the presence of a catalyst System such as palladium (II) dicyclohexyl-[2(2,6-diisopropoxyphenyl)phenyl]phosphane methanesulfonate N-methyl-2-phenyl-aniline and a base such as CS2CO3. The reaction may be performed in the presence of added heat (e.g., 100 °C). In some embodiments, the reaction is performed in a solvent such as toluene. Any suitable method for the removal of an alcohol protecting group may be used to préparé a compound of formula 3-4. For example, where PG² is THP, an acid such as p-toluene sulfonic acid in a solvent such as methanol may be used. The reaction may be performed at room température.

143

Scheme 3

3-2

—- ^R,-pr^

3-4

Scheme 4 shows processes for the préparation of alcohols of formula 4-5 from aryl halides of formula 3-1. Any suitable reagent for performing a lithium-halogen exchange on an 5 heteroaryl bromide, such as treatment with n-butyl lithium, may be used to generate a heteroaryl organometallic reagent in situ. The reaction may be performed in a solvent such as THF or diethyl ether at low température (e.g., 0 to -78 °C). Addition of the organometallic reagent to an epoxide such as ethylene oxide in the presence of a Lewis acid such as trifluoroboron diethyl etherate affords alcohols of formula 4-2. In some embodiments, the lithium halogen exchange 10 reaction may be performed under continuous flow conditions.

In an alternative process for the préparation of compounds of formula 4-2, an aldéhyde of formula 4-3 may undergo a Wittig reaction with a reagent such as an ylide of formula 4-4 to afford an enol ether of formula 4-5. In some embodiments, the reaction is performed in the presence of a base such as potassium tert-butoxide in a solvent such as diethyl ether. In some 15 embodiments, enol ethers of formula 4-5 may be converted to compounds of formula 4-6 by treatment with an acid such as HCl. In some embodiments, a compound of formula 4-2 may be prepared from a compound of formula 4-6 using any suitable reagent for réduction of an aldéhyde to an alcohol, for example, sodium borohydride in methanol may be used.

144

Scheme 4

3-1 4-2

Λ

4-5

Scheme 5 shows processes for the préparation of compounds of formula 1-1. PG³ is any suitable nitrogen protecting group. Compounds of formula 5-1 may be protected with any suitable nitrogen protecting group. For example, where PG³ is a Boc group, any suitable reagents for addition of a Boc group onto an amine may be used. A compound of formula 5-3 (Weinreb amide) may be prepared from a compound of formula 5-2 and N-methyl N-methoxy amine using any suitable amide coupling reagent. For example, the reaction may be performed in a solvent such as dichloromethane in the presence of T3P and DIPEA. A compound of formula 5-5 may be 10 prepared from a compound of formula 5-3 by addition of an organometallic reagent such as methyl magnésium iodide. The reaction may be performed in a solvent such as THF at low température (e.g., 0 °C). Compounds of formula 1-1 may be prepared from compounds of formula 5-5 using any suitable method for the removal of a nitrogen protecting group. For example, where PG³ is Boc, a solution of HCl in 1,4-dioxane may be used.

145

Scheme 5

1-1

Scheme 6 shows an alternative process for the préparation of a compound of formula 1-3 from N-protected beta-amino acids of formula 6-1. PG⁴ may be Boc or any suitable nitrogen protecting group. Compound 6-2 dimagnesium sait may be coupled to compounds of formula 6-1 using a reagent such as CDI in a solvent such as THF. Condensation of compounds of formula 6-3 with aldéhydes of formula 6-4 affords compounds of formula 6-5. In some embodiments, the reaction may be performed by treatment of a compound of formula 6-3 with an acid such as TFA in a solvent such as dichloromethane, followed by the addition of aldéhyde of formula 6-4. A compound of formula 1-3 may be prepared from a compound of formula 6-5 by treatment with an acid such as methanesulfonic acid in a solvent such as dichloromethane. The reaction may be performed in the presence of added heat (e.g., reflux conditions).

146

Préparation of SI

2-(3-thienyl)ethanol (SI)

S1

2-(3-thienyl)ethanol (SI) was obtained from commercial sources.

147

Préparation of S2

2-(5-chloro-3-thienyl)ethanol (S2)

Step 1. Synthesis of tert-butyl-dimethyl-[2-(3-thienyl)ethoxy]silane (Cl)

To a solution of 2-(3-thienyl)ethanol SI (18 g, 140.4 mmol) in DMF (100 mL) was added imidazole (12 g, 176.3 mmol) and teri-butyl-chloro-dimethyl-silane (24 g, 159.2 mmol) sequentially. An exotherm was observed. The reaction mixture was stirred at room température for 3 hours. The reaction mixture was diluted with MTBE (500 mL) and washed with water (200 mL), 0.5 N HCl (200 mL), water (200 mL), and brine (200 mL). The organic layer was dried, filtered, and concentrated in vacuo. The organic layer was dissolved in heptane and passed through a silica gel plug; which was washed with 1-5% MTBE/Heptane. Solvent was removed to afford tert-butyl-dimethyl-[2-(3-thienyl)ethoxy]silane Cl (34 g, 99%). *H NMR (400 MHz, Chloroformé) δ 7.28 - 7.13 (m, 1H), 7.04 - 6.91 (m, 2H), 3.80 (t, J= 6.9 Hz, 2H), 2.90 - 2.75 (m, 2H), 0.88 (s, 9H), -0.00 (s, 6H).

Step 2. Synthesis of tert-butyl-[2-(5-chloro-3-thienyl)ethoxy] -dimethyl-silane (C2)

To a solution of 2,2,6,6-tetramethylpiperidine (36 mL, 213.3 mmol) in tetrahydrofuran (200 mL) cooled to 0 °C was added a solution of hexyllithium (92 mL of 2.3 M, 211.6 mmol). The reaction was stirred for 30 minutes at -78 °C. A solution of ier/-butyl-dimethyl-[2-(3thienyl)ethoxy]silane Cl (34 g, 138.8 mmol) in THF (150 mL) was added to the reaction over 20 minutes. The reaction was stirred at -30 °C for 45 minutes. The reaction was cooled to -78 °C and 1,1,1,2,2,2-hexachloroethane (54 g, 228.1 mmol) was added portion-wise. The reaction was warmed to room température and stirred ovemight. The reaction was quenched with saturated ammonium çhloride (125 mL), diluted with water (100 mL), extracted with EtOAc (500 mL), and back extracted with EtOAc (100 mL). The combined organic layers were washed with 0.5 N HCl (200 mL), water (300 mL), and brine (200 mL). The organic layer was dried over sodium

148 sulfate, filtered, and concentrated to afford the crude product tert-butyl-[2-(5-chloro-3thienyl)ethoxy]-dimethyl-silane C2.

Step 3. Synthesis of 2-(5-chloro-3-thienyl)ethanol (S2)

To a solution of tert-butyl-[2-(5-chloro-3-thienyl)ethoxy]-dimethyl-silane C2 (12.5 g, 42.89 mmol) in 2-Me-THF (120 mL) was added TBAF (63 mL of 1 M in THF, 63.00 mmol). The reaction was stirred at room température ovemight. The reaction was partitioned between EtOAc (400 mL) and water (400 mL). The layers were separated, and the organic layer was extracted with EtOAc (200 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-50% EtOAc in heptane) yielded the product 2-(5-chloro-3-thienyl)ethanol S2 (4.5 g, 58%). *H NMR (300 MHz, Chloroform-<7) δ 6.82 (d, J= 0.9 Hz, 2H), 3.89 - 3.71 (m, 2H), 2.79 (t, J= 6.4 Hz, 2H), 2.05 (s, 1H). LCMS m/z 162.91 [M+H]⁺.

Préparation of S3

2-[5-(trifluoromethyl)-3-thienyl]éthanol (S3)

Step 1. Synthesis of 2-[2-[5-(trifluoromethyl)-3-thienyl] ethoxy] tetra hydropyrane (C5) To a mixture of 4-bromo-2-(trifluoromethyl)thiophene C3 (9 g, 38.96 mmol), dicyclohexyl-[2-(2,6-diisopropoxyphenyl)phenyl]phosphane; methanesulfonate; N-methyl-2phenyl-aniline palladium (2+) (1.8 g, 2.117 mmol), and potassium trifluoro(2-tetrahydropyran-2yloxyethyl)boranuide C4 (10 g, 42.36 mmol) was added toluene (75 mL) and water (25 mL). Nitrogen was passed over the top of the reaction before addition of CS2CO3 (40 g, 122.8 mmol). A reflux condenser was added, and the reaction was heated at 100 °C for 48 hours. The reaction was diluted with EtOAc (150 mL) and water (100 mL). The two layers were separated and the aqueous layer was extracted with EtOAc (100 mL). The combined organics were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-20% EtOAc in heptane) yielded the product 2-[2-[5(trifluoromethyl)-3-thienyl]ethoxy]tetrahydropyran C5 (9 g, 82%). 'H NMR (300 MHz, 149

Chloroform-J) δ 7.37 (t, J= 1.3 Hz, 1H), 7.22 (d, J= 1.5 Hz, 1H), 4.62 (dd, J= 4.2, 2.8 Hz, 1H), 3.96 (dt, J= 9.6, 6.7 Hz, 1H), 3.75 (ddd, J= 11.3, 8.0, 3.4 Hz, 1H), 3.62 (dt, J= 9.6, 6.5 Hz, 1H), 3.55 - 3.41 (m, 1H), 2.93 (t, J= 6.6 Hz, 2H), 1.83 (ddd, J= 14.2, 6.6, 3.4 Hz, 1H), 1.73 (td, J= 9.0, 4.2 Hz, 1H), 1.66 - 1.50 (m, 4H).

Step 2. Synthesis of 2-[5-(trifluoromethyl)-3-thienyl]éthanol (S3)

To a stirred solution of 2-[2-[5-(trifluoromethyl)-3-thienyl]ethoxy]tetrahydropyran C5 (1.8 g, 6.100 mmol) in MeOH (25 mL) was added 4-methylbenzenesulfonic acid monohydrate (1.2 g, 6.309 mmol) at room température. The reaction mixture was stirred at room température for 1 hour. The reaction mixture was diluted with water (100 mL) and extracted with MTBE (2 x 100 mL). The combined organic layers were washed with dilute NaHCCh (10 mL NaHCO₃ and 10 mL water) and brine (10 mL), dried over sodium sulfate, filtered, and evaporated under vacuum to get crude compound. Purification by silica gel chromatography (Gradient: 0-30% EtOAc in heptane) yielded the product 2-[5-(trifluoromethyl)-3-thienyl]ethanol S3 (820 mg, 69%). ‘H NMR (400 MHz, Chloroform-iZ) δ 7.35 (p, J= 1.3 Hz, 1H), 7.23 (dt, J= 1.7, 0.9 Hz, 1H), 3.85 (td, J= 7.1, 6.5, 2.7 Hz, 2H), 2.87 (td, J= 6.4, 0.8 Hz, 2H), 2.06 (d, J= 4.3 Hz, 1H).

Alternative Préparation of S3

2-[5-(trifluoromethyl)-3-thienyl]éthanol (S3)

C3 BF₃0Et₂ S3

A solution of 4-bromo-2-(trifhioromethyl)thiophene C3 (50.13 g, 217.0 mmol) in Et₂O (500 mL) was cooled to -78 °C and nBuLi (91 mL of 2.48 M, 225.7 mmol) was added at a rate adapted to keep the température below -68 °C. The reaction was stirred for 20 minutes and ethylene oxide (14 g, 317.8 mmol) was added at a rate to keep the température below -70 °C. BF3.OEÎ2 (28 mL, 226.9 mmol) was added at a rate to keep the température below -68 °C. The BF3.OEÎ2 addition was highly exothermic. The reaction was stirred for one hour at -78 °C and then poured into 500 mL of 1 N HCl and extracted with 500 mL of Et₂O. The extract was dried with MgSCri, filtered, and evaporated in vacuo. Purification by column chromatography (1600 g: isocratic gradient:10% CH3CN-DCM) afforded 2-[5-(trifluoromethyl)-3-thienyl]ethanol S3 (22.48 g, 53%). ’H NMR (300 MHz, Chloroform-i/) δ 7.36 (t, J = 1.3 Hz, 1H), 7.24 (d, J = 1.5

150

Hz, 1H), 3.88 (q, J = 6.0 Hz, 2H), 2.90 (t, J = 6.3 Hz, 2H), 1.55 (t, J = 5.4 Hz, 1H) ppm. 19F

NMR (282 MHz, Chloroform-t/) δ -55.36 ppm.

Préparation of S4

-ethy 1-3-thienyl)éthanol (S4)

Step 1. Synthesis of 5-bromothiophene-3-carbaldehyde (C7)

To a stirred solution of thiophene-3-carbaldehyde C6 (50 g, 40.717 mL, 0.4458 mol) in DMF (500 mL) was added NBS (119.02 g, 0.6687 mol) at 0 °C. The reaction mixture was stirred at room température for 16 hours. The reaction mixture was quenched with ice cold water (600 mL) and extracted with EtOAc (2 x 600 mL). The combined organic layers were dried over Na2SÜ4, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-2% EtOAc in petroleum ether) yielded the product 5-bromothiophene-3-carbaldehyde C7 (39.2 g, 44%). ‘H NMR (400 MHz, Chloroform-i/) δ 9.77 (s, 1H), 7.99 (d, J=1.2 Hz, 1H), 7.505 (d, J =1.6 Hz, 1H).

Step 2. Synthesis of 2-bromo-4-[(E)-2-methoxyvinyl]thiophene (C8)

To a stirred solution of (methoxymethyl)triphenylphosphonium chloride (115.1 g, 0.3358 mol) in diethyl ether (450.00 mL) at 0 °C was added potassium te/7-butoxide (1 M in THF) (381 mL of 1 M, 0.3810 mol) dropwise. The reaction was stirred at 0 °C for 1 hour. A solution of 5bromothiophene-3-carbaldéhyde C7 (45 g, 0.2215 mol) in diethyl ether (90 mL) was added, and 151 then the reaction mixture was stirred at room température for 30 minutes. The reaction mixture was quenched with NH4CI solution (900 mL) at 0 °C and extracted with EtOAc (2 x 700 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated. Purification by silica gel chromatography (Eluent: petroleum ether) afforded the product, 2-bromo-4-[(E)-2methoxyvinyl]thiophene C8 (44.1 g, 82%). 'H NMR (400 MHz, Chloroform-i/) δ 7.25 (d, J = 2 Hz, 1H), 7.18 (d, J= 0.8 Hz, 1H), 7.00 (d, 7 = 1.8 Hz, 1H), 6.91 (d, J= 12.8 Hz, 1H), 6.97 (d, J =1.2 Hz, 1H), 6.05 (d, J=6.8Hz, 1H), 5.72 (d, .7= 12.8 Hz, 1 H), 5.22 (d, J=6.4Hz, 1H), 3.77 (d, J= 2.8 Hz, 3H), 3.64 (d, J=5.2 Hz, 3H). NMR shows a 1:1 mixture of E and Z isomers.

Step 3. Synthesis of2-(5-bromo-3-thienyl)acetaldehyde (C9)

To a stirred solution of 2-bromo-4-[(£)-2-methoxyvinyl]thiophene C8 (14.1 g, 0.0602 mol) in 1,4-Dioxane (141.00 mL) was added HCl (60.200 mL of 4 M in Dioxane, 0.2408 mol) at 0 °C. The reaction mixture was stirred at room température for 30 minutes. The reaction mixture was quenched with saturated NaHCCL at 0 °C and extracted with EtOAc. The organic layer was dried over Na2SO4, filtered, and concentrated to afford 2-(5-bromo-3-thienyl)acetaldehyde C9 (13.1 g, 89%). Ή NMR (400 MHz, Chloroform-7) δ 9.72 (t, J= 2.4 Hz, 1H), 7.04 (s, 1H), 6.94 (d, J= 1.2 Hz, 1H), 3.66 (d, J= 1.6 Hz, 2H).

Step 4. Synthesis of 2-(5-bromo-3-thienyl)ethanol (CIO)

To a stirred solution of 2-(5-bromo-3-thienyl)acetaldehyde C9 (38.5 g, 0.1524 mol) in MeOH (390 mL) was added NaBH4 (13.3 g, 0.3515 mol) at 0 °C. The reaction was stirred for 1 hour. The reaction mixture was quenched with ice water (400 mL) and concentrated in vacuo to remove the MeOH. The crude residue was diluted with water (500 mL) and extracted with EtOAc (3 x 300 mL). The separated organic layers were dried over Na2SO4, filtered, and concentrated. Purification by column chromatography with neutral alumina (Eluent: 35% EtOAc in petroleum ether) afforded the product 2-(5-bromo-3-thienyl)ethanol CIO (30.2 g, 84%) as a pale yellow liquid. ‘H NMR (300 MHz, DMSO-îZ₆) δ 7.20 (t, J= 0.9 Hz, 1H), 7.10 (d, .7=1.2 Hz, 1H), 4.64 (q, J =5.2 Hz, 1H), 3.59-3.55 (m, 2H), 2.67 (t, J= 6.8 Hz, 2H).

Step 5. Synthesis of 2-[2-(5-bromo-3-thienyl)ethoxy]tetrahydropyran (Cil)

To a stirred solution of 2-(5-bromo-3-thienyl)ethanol CIO (8 g, 0.0328 mol) in THF (80. mL) was added 3,4-dihydro-2H-pyran (3.7696 g, 3.8 mL, 0.0448 mol) and PTSA (259 mg, 0.0015 mol) at room température and then the reaction mixture was stirred at room température for 16 hours. The reaction mixture was quenched with saturated aqueous K2CO3 (300 mL) and extracted with EtOAc (2 x 600 mL). The organic layers were dried over Na2SO4, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-5% EtOAc in petroleum ether) yielded the product 2-[2-(5-bromo-3-thienyl)ethoxy]tetra hydropyran Cil (10.1 g,

152

90%). Ή NMR (400 MHz, Chlorofonn-J) δ 6.95 (d, J= 1.6 Hz, 1H), 6.92 (d, J= 0.8, 1H), 4.59 (t, J=2.8Hz, 1H), 3.94-3.74 (m, 2H), 3.60-3.46 (m, 2H), 2.85 (q, 7=6.4 Hz, 2H), 1.80-1.61 (m, 6H). LCMS m/z 291.03 [M+H]⁺.

Step 6. Synthesis of 2-[2-(5-ethyl-3-thienyl)ethoxy] tetrahydropyran (C12)

To a stirred solution of 2-[2-(5-bromotetrahydrothiophen-3-yl)ethoxy]tetrahydropyran Cil (25 g, 0.0719 mol) in THF (250.00 mL) was added n-BuLi (2.5 M in Hexane) (46.1 mL of 2.5 M, 0.1153 mol) at -76 °C. The reaction was stirred for 1 hour. Ethyl iodide (24.832 g, 12.8 mL, 0.1592 mol) was added at -76 °C and then reaction température was slowly increased to room température, and was then stirred for 16 hours. The reaction mixture was quenched with NH4CI solution (500 mL), and extracted with EtOAc (2 x 300 mL). The combined organic layers were dried over Na₂SÛ4, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-3% EtOAc in petroleum ether) yielded the product 2-[2-(5-ethyl-3thienyl)ethoxy]tetrahydropyran C12 (13.2 g, 59%). LCMS m/z 241.21 [M+H]⁺.

Step 7. Synthesis of 2-(5-ethyl-3-thienyl)ethanol (S4)

To a stirred solution of 2-[2-(5-ethyl-3-thienyl)ethoxy]tetrahydropyran C12 (4.4 g, 0.0142 mol) in MeOH (44 mL) was added PTSA (3.0 g, 0.0174 mol) at room température and the reaction was stirred for 2 hours. The reaction mixture was quenched with saturated NaHCOs solution (150 mL), extracted with EtOAc (2 x 150 mL), dried over Na₂SO4, filtered, and concentrated. Purification by column chromatography with neutral alumina (Eluent: 10% EtOAc in petroleum ether) afforded the product 2-(5-ethyl-3-thienyl)ethanol S4 (1.1 g, 45%). 'H NMR (400 MHz, DMSO-î/ô) δ 6.90 (d, J= 1.2 Hz, 1H), 6.71 (d, 7 = 1.2 Hz, 1H), 4.62-4.58 (m, 1H), 3.59-3.55 (m, 2H), 2.77-2.71 (m, 2H), 2.64 (t, J= 7.2, 2H), 1.22-1.85 (m, 3H).

Préparation of S5

2-(5-ethyl-2-thienyl)ethanol (S5)

Step 1. Synthesis of 2-(5-ethyl-2-thienyl)ethanol (S5)

To a solution of 2-ethylthiophene C13 (54 g, 466.9 mmol) in anhydrous THF (1 L) at 0 °C was added n-BuLi in hexane (255 mL of 2.2 M, 561.0 mmol) over 45 minutes. A light yellow/orange solution resulted. The température range during the addition was 0-10 °C. The 153 mixture was stirred at room température for 30 minutes. After cooling to 0 °C, a solution of ethylene-oxide (200 mL of 2.9 M, 580.0 mmol) was added over 30 minutes. The reaction was stirred at 0 °C for 2 hours and then was warmed to room température. The reaction mixture was quenched with water (700 mL) and saturated NH4CI (200 mL) and the THF was evaporated. The product was extracted with EtOAc (1 x 400 mL; 2 x 150 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The organic layer was passed through a silica gel plug washing with DCM (1000 mL), 80% EtOAc/Heptane (2 x 200 mL), and DCM (2 x 250 mL) to afford 2-(5-ethyl-2-thienyl)ethanol S5 (71.25 g, 93%). *H NMR (300 MHz, Chloroform-J) δ 6.69 (dt, J= 3.4, 0.9 Hz, 1H), 6.64 (dt, J= 3.3, 1.0 Hz, 1H), 3.84 (t, J= 6.3 Hz, 2H), 3.08 - 2.97 (m, 2H), 2.82 (qd, J= 7.5, 1.0 Hz, 2H), 1.31 (t, J= 7.5 Hz, 4H).

Préparation of S6

2-[5-(trifluoromethyl)-2-thienyl]éthanol (S6)

Step 1. Synthesis of 2-(5-iodo-2-thienyl)ethanol (C15)

To a stirred solution of NIS (104.83 g, 0.4680 mol) in DCM (1000 mL) was added 2-(2thienyl)ethanol C14 (50 g, 0.3900 mol) at 0 °C. The reaction was warmed to room température and stirred for 16 hours. The reaction mixture was diluted with DCM (500 mL), washed with saturated sodium thiosulphate, brine, dried over Na2SÜ4, and concentrated in vacuo. Purification by column chromatography (Eluent: 20% EtOAc in petroleum ether) afforded the product 2-(5iodo-2-thienyl)ethanol C15 (62 g, 56%). ‘H NMR (400 MHz, Chloroform-J) δ 7.08 (d, .7 = 3.6 Hz, 1H), 6.57-6.56 (m, 1H), 3.82 (q, J= 6 Hz, 2H), 3.05 (q, J= 6.4 Hz, 2H). LCMS m/z 254.89 [M+H]⁺.

Step 2. Synthesis of 2-[2-(5-iodo-2-thienyl)ethoxy]tetrahydropyran (C16)

To a stirred solution of 2-(5-iodo-2-thienyl)ethanol C15 (15 g, 0.0525 mol) and 3,4-dihydro-2H-pyran (6.6284 g, 0.0788 mol) in THF (60 mL) was added PTSA (1.3604 g,

154

1.2714 mL, 0.0079 mol) at room température. The reaction was stirred for 16 hours under argon balloon pressure. The reaction mixture was concentrated under reduced pressure. Purification by silica gel chromatography (Eluent: 5% EtOAc in petroleum ether) yielded the product 2-[2-(5iodo-2 thienyl)ethoxy]tetrahydropyran C16 (12.8 g, 68%). 'H NMR (400 MHz, DMSO-î/ô) δ 7.14 (d, J= 3.6 Hz, 1H), 6.64 (d, J= 3.6 Hz, 1H), 4.59 (t, J =3.6 Hz, 1H), 3.80-3.76 (m, 1H), 3.74-3.67 (m, 1H), 3.54-3.50 (m, 1H), 3.48-3.41 (m, 1H), 3.03 (t, J= 6 Hz, 2H), 1.75-1.69 (m, 1H), 1.61-1.59 (m, 1H), 1.51-1.42 (m, 4H).

Step 3. Synthesis of 2-[2-[5-(trifluoromethyl)-2-thienyl]ethoxy]tetrahydropyran (Cl 7)

To a stirred solution of 2-[2-(5-iodo-2-thienyl)ethoxy]tetrahydropyran C16 (10 g, 0.0219 mol) and methyl 2,2-difluoro-2-fluorosulfonyl-acetate (12.63 g, 0.0657 mol) in DMF (40 mL) was added copper(I) bromide dimethyl sulfide complex 99% (2.241 g, 0.0109 mol). The reaction was stirred at 100 °C for 16 hours. The reaction was warmed to room température, diluted with EtOAc (100 mL), filtered, and washed with EtOAc (50 mL). The filtrâtes were washed with chilled brine solution, dried over Na2SO4, and concentrated under reduced pressure. Purification by column chromatography with neutral alumina (Eluent: 5% EtOAc in petroleum ether) afforded the product 2-[2-[5-(trifluoromethyl)-2-thienyl]ethoxy]tetrahydro pyran C17 (2.9 g, 41%). ’H NMR (400 MHz, ChlorofomwZ) δ 7.25 (s, 1H), 6.82-6.81(m, 1H), 4.63 (t, J =3.6 Hz, 1H), 4.00-3.95 (m, 1H), 3.78-3.75 (m, 1H), 3.64-3.58 (m, 1H), 3.51-3.48 (m, 1H), 3.12 (d, J = 6.4 Hz, 2H), 1.90-1.80 (m, 1H), 1.73-1.64 (m, 1H), 1.65-1.51 (m, 4H). GCMS: 87.26%, m/z: 280 [M]⁺.

Step 4. Synthesis of 2-[5-(trifluoromethyl)-2-thienyl]éthanol (S6)

To a stirred solution of 2-[2-[5-(trifluoromethyl)-2-thienyl]ethoxy]tetrahydropyran C17 (5.8 g, 0.0170 mol) in MeOH (100 mL) was added PTSA (2.93 g, 0.0170 mol) at room température. The reaction was stirred for 16 hours. The reaction mixture was concentrated under reduced pressure. Purification by column chromatography with neutral alumina (Eluent: 10% EtOAc in petroleum ether) afforded the product 2-[5-(trifluoromethyl)-2-thienyl]ethanol S6 (2.3 g, 61%). ‘H NMR (400 MHz, DMSO-J^ δ 7.52-7.51 (m, 1H), 6.99-6.98 (m, 1H), 4.92 (t, J= 4.8 Hz, 1H), 3.65-3.61 (m, 2H), 2.98 (t, J= 6 Hz, 2H). ¹⁹F NMR (376.22 MHz, DMSO-d₆) δ -53.53 (s, 3F). GCMS: 88.56% m/z\ 196.0 [M]⁺.

155

Préparation ofS7

2-[5-(trifluoromethyl)-2-thienyl]propan-l-ol (S7)

Step 1. Synthesis of ethyl 2-(2-thienyl)acetateethanol (C19)

To a stirred solution of 2-(2-thienyl)acetic acid C18 (100 g, 703.35 mmol) in éthanol (2000 mL) was added HCl (aqueous) (50 mL of 36% (w/v), 493.68 mmol) at room température. The reaction mixture was stirred for 12 hours at 70 °C. The mixture was concentrated, and the resulting crude material was diluted with EtOAc (1000 mL), washed with 5% Na2CÜ3 aqueous solution (3 x 200 mL), and brine (200 mL). The organic layer was dried and concentrated to afford desired product, ethyl 2-(2-thienyl)acetate C19 (100 g, 82%). *H NMR (Chloroform-J, 400 MHz) δ 7.22-7.21 (dd, J= 1.2 Hz, J= 3.6 Hz, 1H), 6.97-6.95 (m, 2H), 4.21-4.16 (q, J= 7.2 Hz, 2H), 3.83 (s, 2H), 1.30-1.26 (t, J= 7.2 Hz, 3H). LCMS m/z 171.26 [M+H]⁺.

Step 2. Synthesis of ethyl 2-(2-thienyl)propanoate (C20)

To a solution of ethyl 2-(2-thienyl)acetate C19 (1.36 g, 7.99 mmol) in THF (20 mL) at 78 °C was added (diisopropylamino)lithium (8 mL of 1 M, 8.000 mmol). After 15 minutes, Mel (500 pL, 8.032 mmol) was added and the reaction was mixture stirred at -78 °C for 2 hours. The reaction was quenched with saturated NH4CI (50 mL) and extracted with EtOAc. The organic layer was dried and concentrated to an oil. Purification by silica chromatography (Gradient: 0 to 25% EtOAc in heptane) afforded the product, ethyl 2-(2-thienyl)propanoate C20 (1.04 g, 71%). Ή NMR (300 MHz, Chloroformé δ 7.25 - 7.17 (m, 1H), 7.02 - 6.93 (m, 2H), 4.18 (d, J= 7.2 Hz, 2H), 4.02 (q, J= 7.1 Hz, 1H), 1.60 (d, J= 7.2 Hz, 3H), 1.28 (t, J= 7.1 Hz, 3H).

Step 3. Synthesis of ethyl 2-(5-iodo-2-thienyl)propanoate (C21)

To a stirred solution of ethyl 2-(2-thienyl)propanoate C20 (35 g, 143.99 mmol) in Acetic acid (350 mL) was added N-Iodosuccinimide (38.875 g, 172.79 mmol). The reaction mixture was stirred for one hour at 100 °C. The mixture was concentrated and the resulting crude material was diluted with EtOAc (700 mL), washed with water (300 mL), saturated sodium bicarbonate solution (300 mL), saturated sodium thiosulfate solution (300 mL), and brine 156 solution (250 mL) sequentially. The organic layer was dried over NazSCM, filtered, and concentrated to afford crude product. Purification by silica gel chromatography (Eluent: 3% EtOAc in petroleum ether) yielded the product ethyl 2-(5-iodo-2-thienyl)propanoate C21 (30 g, 42%). ‘H NMR (Chloroform-c/, 400 MHz) δ 7.08 (d, J= 4 Hz, 1H), 6.62 (d, J= 4 Hz, 1H), 4.194.13 (m, 2H), 3.98-3.92 (m, 1H), 1.55-1.51 (m, 3H), 1.28-1.24 (m, 3H). LCMS m/z 309.9 [M+H]⁺.

Step 4. Synthesis of ethyl 2-[5-(trifluoromethyl)-2-thienyl]propanoate (C22)

To a stirred solution of ethyl 2-(5-iodo-2-thienyl)propanoate C21 (5 g, 9.9629 mmol) and Methyl 2,2-difhioro-2-(fluorosulfonyl)acetate (9.57 g, 49.814 mmol) in DMF (50 mL) was added Cul (2.2768 g, 11.955 mmol) under nitrogen atmosphère. The reaction mixture was stirred for 12 hours at 100 °C. The mixture was filtered through Celite® and the Celite® pad was washed with Diethyl Ether (2 x 100 mL). Filtrate was quenched with cold water (100 mL). The two layers were separated and the aqueous layer was extracted with diethyl ether (2x50 mL). The combined organic layers were washed with brine (30 mL), dried, and concentrated. Purification by silica gel chromatography (Eluent: 3% EtOAc in petroleum ether) yielded the product ethyl 2-[5-(trifluoromethyl)-2-thienyl]propanoate C22 (2 g, 58%). ’H NMR (Chloroform-J, 400 MHz) δ 7.29-7.26 (m, 1H), 6.92-6.90 (m, 1H), 4.21-4.15 (m, 2H), 3.99-3.96 (m, 1H), 1.57-1.53 (m, 3H), 1.23-1.27(m, 3H). GCMS: m/z\ 252.1 [M] ⁺

Step 5. Synthesis of 2-[5-(trifluoromethyl)-2-thienyl]propan-l-ol (S7)

To a stirred solution of ethyl 2-[5-(trifluoromethyl)-2-thienyl]propanoate C22 (12 g, 41.701 mmol) in THF (250 mL) was added DIBAL-H (35.584 mL of 25% (w/v), 62.5 mmol) dropwise at 0 °C. The reaction mixture was stirred for 2 hours at 0 °C. The mixture was slowly quenched with saturated NH4CI solution (300 mL) at 0 °C and the suspension was filtered through Celite® and the Celite® pad was washed with EtOAc (2 x 200 mL). The filtrate was separated into layers. The aqueous layer was extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SÛ4, and concentrated. Purification by silica gel chromatography (Eluent: 3% EtOAc in petroleum ether) yielded crude product. The racemic compound,2-[5-(trifluoromethyl)-2-thienyl]propan-l-ol (1.6 g, 7.3067 mmol) was separated from the dimethyl over alkylation byproduct using chiral SFC séparation. Column: Daicel Chiralpak ® AD-H, 30 x 250 mm; Mobile Phase: 10% Methanol/Hexane Mixture (7:3), 90% carbon dioxide. Flow: 90 g/minutes. 2-[5(trifluoromethyl)-2-thienyl]propan-l-ol S7 (3.64 g). 'HNMR (400 MHz, Chloroform-<7) δ 7.52 (m, 1H), 7.00 (m, 1H), 4.97 (t, J =5.6 Hz, 1H), 3.51 (t, J =6.0 Hz, 2H) 3.17 (m, 1H), 1.27 (d, J= 6.8 Hz, 3H). GCMS: m/z\ 210.0 [M]⁺.

157

Préparation of S8

2-methyl-2-[5-(trifluoromethyl)-2-thienyl]propan-l-ol (S8)

S8

S8 was obtained during SFC purification of S7 as a side product due to over alkylation in step 2 described above.

Préparation of S9, S10, and SU

2-methyl-2-[5-(chloro)-2-thienyl]propan-l-ol (S9)

2-[5-(chloro)-2-thienyl]propan-l-ol (S10 [ENANT-1], SU [ENANT-2])

Step 1. Synthesis of ethyl 2-(5-chloro-2-thienyl)propanoate (C24)

To a stirred solution of ethyl 2-(2-thienyl)propanoate C20 (1 g, 4.1139 mmol) in acetic acid (10 mL) was added N-Chlorosuccinimide C23 (549.34 mg, 4.1139 mmol). The reaction mixture was stirred for 1 hour at 100 °C. The mixture was concentrated and the resulting crude material was diluted with EtOAc (25 mL), washed with water (10 mL), saturated sodium bicarbonate solution (10 mL), saturated sodium thiosulfate solution (10 mL), and brine solution (10 mL). The organic layer was dried over NaiSCU, filtered, and concentrated to afford crude product. Purification by silica gel chromatography (Eluent: 3% EtOAc in petroleum ether) yielded the product ethyl 2-(5-chloro-2-thienyl)propanoate C24 (700 mg, 60%). *H NMR (Chloroformé, 400 MHz): δ = 6.75-6.73 (m, 1H), 6.71-6.69 (m, 1H), 4.20-4.14 (m, 2H), 3.883.73 (q, 7= 6.4 Hz, 1H), 1.55-1.53 (t, 7=2.8 Hz, 3H), 1.30-1.221 (m, 3H). GCMS: m/z\ 218.0 [M] ⁺

158

Step 2. Synthesis of 2-(5-chloro-2-thienyl)-2-methyl-propan-l-ol and 2-(5-chloro-2thienyl)propan-l-ol (S9) and (C25)

To a stirred solution of ethyl 2-(5-chloro-2-thienyl)propanoate C24 (25 g, 86.877 mmol) in THF (500 mL) was added DIBAL-H (74.135 mL of 25% (w/v), 130.32 mmol) dropwise at 0 °C. The reaction mixture was stirred for 2 hours at 0 °C. The mixture was slowly quenched with saturated NH4CI solution (300 mL) at 0 °C and the suspension was filtered through Celite® and the Celite® pad was washed with EtOAc (2 x 200 mL). The filtrate was separated into two layers. The aqueous layer was extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with brine (200 mL), dried over sodium sulfate, and concentrated. Purification by silica gel chromatography (Eluent: 3% EtOAc in petroleum ether) yielded S9 2-(5-chloro-2thienyl)-2-methyl-propan-l-ol (410 mg, 2%). *H NMR (Chloroformé, 400 MHz) δ 6.76-6.75 (d, J= 4 Hz, 1H), 6.67-6.65(t, J= 4 Hz, 1H), 3.54-3.52 (d, J= 6.8 Hz, 2H), 1.47-1.43 (t, J= 6.8 Hz, 1H), 1.34 (s, 6H). GCMS: m/z\ 190.0 [M]⁺; and 2-(5-chloro-2-thienyl)propan-l-ol C25 (12 g, 72%). Ή NMR (Chloroformé, 400 MHz) δ 6.76-6.75 (d, J= 3.6 Hz, 1H), 6.66-6.65 (dd, 4.4 Hz, 1H), 3.71-3.61 (m, 2H), 3.15-3.10 (m, 1H), 1.57-1.52 (m, 1H), 1.34-1.31 (t, J= 6 Hz, 3H). GCMS: m/z\ 176.0 [M]⁺. NOTE: the dimethyl compound (S9) was formed as a side product due to over alkylation during synthesis of C20.

Step 3. Synthesis of2-(5-chloro-2-thienyl)propan-l-ol (S10) and (SU)

The racemic compound, 2-(5-chloro-2-thienyl)propan-l-ol C25 (12 g, 62.492 mmol) was separated into constituent enantiomers by chiral SFC séparation. Column: Daicel Chiralpak ® AD-H, 30 x 250 mm; Mobile Phase: 10% Methanol/Hexane Mixture (7:3), 90% carbon dioxide. Flow: 90 g/minutes. 2-(5-chloro-2-thienyl)propan-l-ol S10 (4 g, 35%). *H NMR (Chloroformé, 400 MHz) δ 6.76-6.75 (d, J= 3.6 Hz, 1H), 6.66-6.65 (dd, J= 3.6 Hz, 1H), 3.73-3.61 (m, 2H), 3.17-3.10 (m, 1H), 1.52-1.49 (t, J= 5.2 Hz, 1H), 1.32-1.30 (d, J= 6.8 Hz, 3H). GCMS: m/z: 176.0 [M]⁺; and 2-(5-chloro-2-thienyl)propan-l-ol SU (3.75 g, 34%). *H NMR (Chloroformé, 400 MHz) δ 6.76-6.75 (d, J= 4 Hz, 1H), 6.66-6.65 (dd, J= 3.6 Hz, 1H), 3.73-3.61 (m, 2H), 3.15-3.10 (q,J= 6.8 Hz, 1H), 1.51-1.48 (t, J= 5.6 Hz, 1H), 1.33-1.30 (d, J= 7.2Hz, 3H). GCMS: m/z\ 176.0 [M]⁺.

159

Préparation of S12 and S13

Step 1. Synthesis of ethyl 2-(5-acetyl-2-thienyl)propanoate (C26)

To a stirred solution of ethyl 2-(2-thienyl)propanoate C20 (80 g, 336.92 mmol) in DCM (1500 mL) was added Acetyl chloride (39.671 g, 35.934 mL, 505.38 mmol) dropwise at 0 °C, followed by addition of AICh (67.388 g, 505.38 mmol) at 0 °C. The reaction mixture was stirred for 2 hours at 0 °C. The mixture was slowly quenched with ice water (1000 mL), the two layers were separated, and the aqueous layer was extracted with DCM (2 x 500 mL). The combined organic layers were washed with brine (500 mL), dried over sodium sulfate. Purification by silica gel chromatography (Gradient: 0-5% EtOAc in petroleum ether) yielded the product ethyl 2-(5-acetyl-2-thienyl)propanoate C26 (60 g, 73%). 'H NMR (Chloroform-<7, 400 MHz) δ 7.567.54 (t, J= 4.0 Hz, 1H), 6.99-6.98 (m, 1H), 4.20-4.14 (m, 2H), 4.01-3.96 (q, J= 7.2 Hz, 1H), 2.52 (s, 3H), 1.60-1.56 (d, J= 7.2 Hz, 3H), 1.28-1.23 (m, 3H). LCMS m/z 227.1 [M+H]⁺.

Step 2. Synthesis of ethyl 2-(5-ethyl-2-thienyl)propanoate (C27)

To a stirred solution of ethyl 2-(5-acetyl-2-thienyl)propanoate C26 (60 g, 245.79 mmol) in TFA (400 mL) was added Triethyl-silane (42.870 g, 58.9 mL, 368.69 mmol) dropwise at 0 °C. The reaction mixture was stirred for 4 hours at room température. The reaction was concentrated and quenched with ice water (500 mL) and extracted with EtOAc (3 x 500 mL). The combined organic layers were washed with brine (250 mL), dried over sodium sulfate, and concentrated to afford crade product. Purification by silica gel chromatography (Gradient: 0-3% EtOAc in petroleum ether) yielded the product ethyl 2-(5-ethyl-2-thienyl)propanoate C27 (50 g, 82%). ‘H NMR (Chloroform-J, 400 MHz) δ 6.73-6.72 (dd, J= 3.6 Hz, 1H), 6.62-6.60 (m, 1H), 4.18-4.13 (m, 2H), 3.93-3.88 (q, J= 7.2 Hz, 1H), 2.82-2.78 (m, 2H), 1.55-1.53 (d, 7=7.2 Hz, 3H) 1.30-1.23 (m, 6H). LCMS m/z 213.2 [M+H]⁺.

160

Step 3. Synthesis of 2-(5-ethy 1-2-thienyl)propan-l-ol (C28)

To a stirred solution of ethyl 2-(5-ethyl-2-thienyl)propanoate C27 (50 g, 200.18 mmol) in THF (1000 mL) was added DIBAL-H (25% in toluene) (227.75 mL of 25% (w/v), 400.36 mmol) dropwise at 0 °C. The reaction mixture was stirred for 2 hours at 0 °C. The mixture was slowly quenched with saturated NH4CI solution (500 mL) at 0 °C and extracted with EtOAc (2 x 500 mL). The combined organic layers were washed with brine (250 mL), dried over sodium sulfate, and concentrated. Purification by silica gel chromatography (Gradient: 0-5% EtOAc in Petroleum ether) yielded the product 2-(5-ethyl-2-thienyl)propan-l-ol C28 (31 g, 89%). 'H NMR (Chloroform-e/, 400 MHz): δ 6.69-6.68 (d, J= 3.6 Hz, 1H), 6.64-6.62 (m, 1H), 3.72-3.60 (m, 2H), 3.18-3.13 (q, J= 6.8 Hz, 1H), 2.83-2.77 (m, 2H),1.61-1.5 (m, 1H), 1.35-1.28 (m, 6H). LCMS m/z 171.02 [M+H]⁺.

Step 4. Synthesis of2-(5-ethyl-2-thienyl)propan-l-ol (S12) and (S13)

The racemic compound 2-(5-ethyl-2-thienyl)propan-l-ol C28 (31 g, 178.06 mmol) was separated into constituent enantiomers by chiral SFC séparation. Column: Daicel Chiralpak ® AD-H, 30 x 250 mm; Mobile Phase: 10% Methanol/Hexane Mixture (7:3), 85% carbon dioxide. 2-(5-ethyl-2-thienyl)propan-l-ol S12 (13.45 g, 43%). ’H NMR (Chloroform-<7, 400 MHz): δ = 6.69-6.68 (d, 3.2 Hz, 1H), 6.63-6.62 (d, J= 3.2 Hz, 1H), 3.73-3.61 (m, 2H), 3.19-3.14 (q, J=

6.8 Hz, 1H), 2.83-2.78 (m, 2H), 1.54-1.47 (m, 1H), 1.35-1.27 (m, 6H). LCMS m/z 171.1 [M+H]⁺; And 2-(5-ethyl-2-thienyl)propan-l-ol S13 (11.35 g, 37%). Ή NMR (Chloroform-J, 400 MHz): δ 6.68-6.67 (d, J= 3.6 Hz, 1H), 6.63 (d, J= 3.6 Hz, 1H), 3.73-3.61 (m, 2H), 3.203.12 (m, 1H), 2.83-2.77 (q, J= 7.6 Hz, 2H), 1.54-1.45 (m, 1H), 1.33-1.27 (m, 6H). LCMS m/z 171.1 [M+H]⁺.

Préparation of S14

2-(5-methyl-3-thienyl)ethanol (S14)

C10 Xphos Pd G2 S14

To a stirred solution of 2-(5-bromo-3-thienyl)ethanol CIO (2.5 g, 0.0098 mol) in 1,4Dioxane (16.000 mL) was added K2CO3 (4.9 g, 0.036 mol) at room température in a sealed tube. The reaction mixture was degassed with argon gas for 10 minutes. Xphos Pd G2 (457 mg, 580.83 pmol) was added and again degassed for 5 minutes. Trimethylboroxine (50% solution in THF) (24.605 mL of 50% (w/v), 0.0980 mol) was added and heated to 80 °C for 16 hours. The 161 reaction mixture was diluted with water (200 mL) and extracted with EtOAc (3 x 150 mL). The combined organic layers were dried over Na2SÜ4, filtered, and concentrated under reduced pressure. Purification by column chromatography (Eluent: 20% EtOAc in petroleum ether) afforded the product S14 2-(5-methyl-3-thienyl)ethanol (950 mg, 66%) as a yellow liquid. 'H NMR (400 MHz, DMSO-î7₆) δ 6.87 (d, J= 0.8 Hz, 1H), 6.68 (s, 1H), 4.59 (t, J= 5.2 Hz, 1H), 3.58-3.53 (m, 2H), 2.63 (t, J= 7.2 Hz, 2H), 2.38 (d, J= 0.8 Hz, 3H).

Préparation ofS15

2-(5-methyl-2-thienyl)ethanol (S15)

N BS

Step 1. Synthesis of 2-(5-bromo-2-thienyl)ethanol (C30)

A solution of 2-(2-thienyl)ethanol C29 (15 g, 0.1170 mol) in DMF (150.00 mL) was added dropwise to a solution of NBS (20.824 g, 0.1170 mol) in DMF at -10 °C. The reaction was stirred at room température for 16 hours. The reaction mixture was quenched with water (300 mL) and extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with 6% KOH solution, ice water (2 x 150 mL), and brine (150 mL). The organic layer was dried over sodium sulfate and concentrated. Purification by column chromatography (Eluent: 10% EtOAc in petroleum ether) afforded the product 2-(5-bromo-2-thienyl)ethanol C30 (20.5 g, 79%). *H NMR (400 MHz, Chloroform-J) δ 6.89 (d, J= 3.6 Hz, 1H), 6.64 - 6.28 (m, 1H), 3.82 (t, J= 6.0 Hz, 2H), 2.99 (t, J= 6.0 Hz, 2H).

Step 2. Synthesis of 2-[2-(5-bromo-2-thienyl)ethoxy]tetrahydropyran (C31)

To a stirred solution of 2-(5-bromo-2-thienyl)ethanol C30 (20 g, 0.0869 mol) and 3,4dihydro-2H-pyran (10.969 g, 0.1304 mol) in THF (80 mL) was added with PTSA (603 mg, 0.5636 mL, 0.0035 mol) and reaction was stirred for 24 hour at room température. The reaction mixture was diluted with EtOAc, washed with saturated sodium bicarbonate solution (50 mL), water, and brine. The organic layer was separated, dried over sodium sulfate, and concentrated. Purification by silica gel chromatography (Gradient: 0-5% EtOAc in petroleum ether) yielded 2162

[2-(5-bromo-2-thienyl)ethoxy]tetrahydropyran C31 (18.5 g, 64%). *H NMR (400 MHz, Chloroform-iZ) δ 6.86 (d, J= 3.6 Hz, 1H), 6.61-6.60 (m, 1H), 4.62 (t, J= 3.6 Hz, 1H), 3.99-3.50 (m, 4H), 3.05-3.01 (m, 2H), 1.73-1.50 (m, 6H).

Step 3. Synthesis of 2-[2-(5-methyl-2-thienyl)ethoxy] tetrahydropyran (C32)

To a solution of 2-[2-(5-bromo-2-thienyl)ethoxy]tetrahydropyran C31 (19 g, 0.0555 mol) in THF (380.00 mL) was added n-BuLi (33.320 mL of 2.5 M, 0.0833 mol) dropwise at 78 °C. The reaction was stirred for one hour at -78 °C. lodomethane (15.755 g, 6.9101 mL, 0.1110 mol) was added dropwise at -78 °C and the reaction mixture was allowed to stir at room température for 16 hours. The reaction mixture was quenched with saturated NH4CI solution and diluted with water. The aqueous layer was extracted with EtOAc (2 x 250 mL). Purification by silica gel chromatography (Eluent: 100% petroleum ether) yielded 2-[2-(5-methyl-2thienyl)ethoxy]tetrahydropyran C32 (19 g, 130%). 'H NMR (400 MHz, Chloroform-<7) δ 6.61 (d, J= 3.2 Hz, 1H), 6.55-6.54 (m, 1H), 4.63 (m, 1H), 3.96-3.50 (m, 4H), 3.03 (t, J= 2.8 Hz, 2H), 2.42 (s, 3H), 1.72-1.42 (m, 6H). .

Step 4. Synthesis of 2-(5-methyl-2-thienyl)ethanol (S15)

To a solution of 2-[2-(5-methyl-2-thienyl)ethoxy]tetrahydropyran C32 (14 g, 0.0532 mol) in MeOH (280.00 mL) was added with PTSA (10.9 g, 10.187 mL, 0.0633 mol) at room température. The reaction was stirred for 24 hours. The reaction mixture was diluted with EtOAc (500 mL) and then washed with water (200 mL). The organic layer was washed with saturated aqueous sodium bicarbonate solution (2 x 100 mL). Aqueous layer was again extracted with EtOAc (2 x 100 mL). The combined organic layers were dried over Na2SO4. Purification by silica gel chromatography (Gradient: 0-15% EtOAc in petroleum ether) yielded 2-(5-methyl-2thienyl)ethanol S15 (6.56 g, 82%). Ή NMR (400 MHz, DMSO-cfo) δ 6.61 (d, J= 3.6 Hz, 1H), 6.58-6.57 (d, J= 4.0 Hz, 1H), 4.73 (t, J= 5.2 Hz, 1H), 3.58-3.53 (m, 2H), 2.82 (t, J= 6.8 Hz, 2H), 2.36 (s, 3H).

163

Préparation ofS16

[4-(2-hydroxyethyl)-2-(trifluoromethyl)-3-thienyl]methyl acetate (S16)

Step 1. Synthesis of tert-butyl-dimethyl-[2-[5-(trifluoromethyl)-3-thienyl]ethoxy]silane (C33)

To a mixture of 2-[5-(trifluoromethyl)-3-thienyl]ethanol S3 (500 mg, 2.498 mmol) in DCM (10 mL) was added imidazole (190 mg, 2.791 mmol) followed by TBSC1 (420 mg, 2.787 mmol) which immediately precipitated a white solid. The solid was filtered and the organic layer was washed with 1 N HCl (10 mL), brine (10 mL), dried with MgSC>4, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-30% EtOAc in heptane) yielded the product, tert-butyl-dimethyl-[2-[5-(trifluoromethyl)-3-thienyl]ethoxy]silane C33, assumed to be quantitative and carried forward without further purification.

Step 2. Synthesis of tert-butyl-dimethyl-[2-[5-(trifluoromethyl)-2-trimethylsilyl-3thienyl]ethoxy]silane (C34)

A mixture of tert-butyl-dimethyl-[2-[5-(trifluoromethyl)-3-thienyl]ethoxy]silane C33 in THF (10 mL) was cooled to -78 °C and sec-butyllithium (2.3 mL of 1.4 M, 3.220 mmol) was added followed by TMSC1 (3 mL of 1 M, 3.000 mmol). After 5 minutes, the yellow mixture was quenched with saturated aqueous ammonium chloride. The mixture was diluted with water (10 mL) and MTBE (10 mL). The organic layer was washed with brine, dried with MgSÛ4, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-10% EtOAc in heptane) yielded Z‘eri-butyl-dimethyl-[2-[5-(trifluoromethyl)-2-trimethylsilyl-3-thienyl]ethoxy]silane C34 (400 mg, 42%). ’H NMR (300 MHz, Chloroform-rf) δ 7.41 (d, J= 1.2 Hz, 1H), 3.80- 3.75 (m, 2H), 2.87 (t, J= 6.8 Hz, 2H), 0.87 (s, 9H), 0.36 (s, 9H), -0.00 (d, J= 2.2 Hz, 6H).

164

Step 3. Synthesis of 4-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-(trifluoromethyl)-5trimethylsilyl-thiophene-3-carbaldehyde (C35)

To a mixture of ler/-butyl-dimethyl-[2-[5-(trifluoromethyl)-2-trimethylsilyl-3thienyl]ethoxy]silane C34 (400 mg, 1.024 mmol) in THF (10 mL) cooled to -78 °C was added sec-butyllithium (1.2 mL of 1.4 M, 1.680 mmol) followed by DMF(3 mL of 1 M, 3.000 mmol). After 5 minutes, the yellow mixture was quenched with saturated aqueous ammonium chloride. The mixture was diluted with EtOAc (20 mL) and water (20 mL) and separated. The organic layer was washed with brine (20 mL), dried with MgSO4, fïltered, and concentrated. Purification by silica gel chromatography (Eluent: 100% heptane) yielded the product, 4-[2-[tertbutyl(dimethyl)silyl]oxyethyl]-2-(trifluoromethyl)-5-trimethylsilyl-thiophene-3-carbaldehyde C35. The mixture was concentrated, diluted with heptane (5 mL) and washed with water (5 mL). The organic layer was passed over a phase separator, concentrated, and telescoped directly to the next step.

Step 4. Synthesis of [4-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-(trifluoromethyl)-5trimethylsilyl-3-thienyl]methanol (C36)

4-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-(trifluoromethyl)-5-trimethylsilyl-thiophene3-carbaldehyde C35 was diluted in MeOH (1 mL) and to the mixture was added NaBHj (7 mg, 0.1850 mmol). After 10 minutes the mixture was concentrated, and re-diluted in heptane (2 mL) and water (2 mL). The organic layer was separated and the aqueous layer was extracted with additional heptane. The organic layer was passed over a phase separator and concentrated. Purification by silica gel chromatography (Gradient: 0-10% EtOAc in heptane) yielded the product C36. *H NMR (300 MHz, Chloroform-d) δ 4.65 (d, J= 6.3 Hz, 2H), 4.00 - 3.72 (m, 2H), 3.34 (t, J= 6.3 Hz, 1H), 2.97 (t, J= 6.1 Hz, 2H), 0.82 (s, 10H), 0.36 (s, 9H).

Step 5. Synthesis of [4-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-(trifluoromethyl)-5trimethylsilyl-3-thienyl]methyl acetate (C37)

To [4-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-(trifluoromethyl)-5-trimethylsilyl-3thienyl]methanol C36 in DCM (4 mL) was added DMAP (2 mg, 0.016 mmol) and DIPEA (50 pL, 0.2871 mmol) followed by Ac2O (30 pL, 0.3180 mmol). The mixture was concentrated, diluted with heptane (5 mL) and washed with water (5 mL). The organic layer was passed over a phase separator and concentrated to yield the product which was telescoped directly in to the next step.

Step 6. Synthesis of [4-(2-hydroxyethyl)-2-(trifluoromethyl)-3-thienyl]methyl acetate (S16) [4-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-(trifluoromethyl)-5-trimethylsilyl-3thienyl]methyl acetate C37 from step 5 was diluted with EtOAc (2 mL) and to the mixture was 165 added a THF solution of TBAF (1 mL of 1 M, 1.000 mmol) and the mixture was stirred. The reaction was stirred for 48 hours. The mixture was diluted with additional EtOAc (3 mL), washed with water, passed over a phase separator, and concentrated. Purification by silica gel chromatography (Gradient: 0-60% EtOAc in heptane) yielded the product [4-(2-hydroxyethyl)-2(trifhioromethyl)-3-thienyl]methyl acetate S16 (35 mg, 12%). 'H NMR (300 MHz, Chloroformd) δ 7.26 (s, 1H), 5.14 (d, J= 1.1 Hz, 2H), 3.86 (t, J= 6.4 Hz, 2H), 2.97 - 2.74 (m, 2H), 2.07 (s, 3H), 1.80 (s, 1H). LCMS m/z 269.21 [M+H]⁺.

Préparation of SI 7 l-methyltriazole-4-carbaldehyde (SI 7)

S17 l-methyltriazole-4-carbaldehyde S17 was obtained from commercially available sources.

Préparation ofS18 l-(2-methylsulfonylethyl)triazole-4-carbaldehyde (S18)

Step 1. Synthesis of l-azido-2-methylsulfonyl-ethane (C40)

A solution of 2-methylsulfonylethanol C38 (5 g, 0.04 mol) and Diphenyl phosphoryl azide C39 (8.8614 g, 0.0322 mol) in Toluene (50 mL) was stirred at 0 °C for 10 minutes and DBU (5.5 g, 5.42 mL, 0.04 mol) was added dropwise at 0 °C over 10 minutes and the reaction was stirred at room température for 16 hours. The reaction mixture was quenched with water (25 mL) and EtOAc (100 mL) and stirred for 20 minutes. The organic layer was separated and aqueous layer was again extracted with EtOAc (2 x 100 mL). The organic layer was dried over Na2SO4 and concentrated. Purification by silica gel chromatography (Gradient: 0-100% ethyl acetate in petroleum ether) gave l-azido-2-methylsulfonyl-ethane C40 (5.2 g, 86%). *H NMR (400 MHz, DMSOé) δ 3.77-3.73 (t, J= 8.8 Hz, 2H), 3.44-3.42 (t, J= 8.8 Hz, 2H), 3.03 (s, 3H).

Step 2. Synthesis of l-(2-methylsulfonylethyl)triazole-4-carbaldehyde (S18)

166

A mixture of 3,3-diethoxyprop-l-yne (555 pL, 3.897 mmol), l-azido-2-methylsulfonylethane C40 (600 mg, 4.022 mmol), CuSO4 (15 mg, 0.09398 mmol), l-(l-benzyltriazol-4-yl)N,N-bis[(l-benzyltriazol-4-yl)methyl]methanamine (100 mg, 0.1885 mmol), and sodium ascorbate (700 mg, 3.974 mmol) in MeOH (12 mL)/water (3 mL) was heated to 60 °C for 2 hours. The reaction was cooled to room température, concentrated, and diluted in EtOAc (100 mL) and water (50 mL). The layers were split and the aqueous layer was extracted with EtOAc (50 mL). The layers were combined and dried, diluted in 1 N HCl (20 mL), and stirred ovemight. At this time, the solution was concentrated to yield l-(2-methylsulfonylethyl)triazole-4carbaldehyde (Hydrochloride sait) S18 (553 mg, 59%). *H NMR (400 MHz, Methanol-iL) δ 8.07 (s, 1H), 5.58 - 5.45 (m, 1H), 4.89 - 4.82 (m, 2H), 3.76 - 3.67 (m, 2H), 3.24 (s, 3H). LCMS m/z 204.47 [M+H]⁺.

Préparation ofS19 l-(2-methylsulfonylethyl)pyrazole-4-carbaldehyde (SI9)

A solution of lH-pyrazole-4-carbaldehyde C42 (10 g, 104.1 mmol) 11methylsulfonylethylene C41 (10 mL, 114.2 mmol) and K2CO3 (25 g, 180.9 mmol) in THF (200 mL) was stirred at 60 °C. After stirring ovemight, the mixture was cooled to room température and concentrated to dryness. The product was suspended in diethyl ether (100 mL) to triturate the product and stirred for 2 hours. The product was filtered and dried ovemight to yield 11-(2methylsulfonylethyl)pyrazole-4-carbaldehyde S19 (20.28 g, 83%). *H NMR (400 MHz, DMSOd₆) δ 9.80 (s, 1H), 8.54 (d, J= 0.7 Hz, 1H), 8.05 (d, J= 0.7 Hz, 1H), 4.64 (t, J= 6.8 Hz, 2H), 3.80 - 3.67 (m, 2H), 2.96 (d, J= 0.7 Hz, 3H). LCMS m/z 203.01 [M+H]⁺.

Préparation of S20 l-[2-[tert-butyl(dimethyl)silyl]oxyethyl]pyrazole-4-carbaldehyde (S20)

Step 1. Synthesis of tert-butyl-(2-iodoethoxy)-dimethyl-silane (C44)

To a stirred solution of 2-iodoethanol C43 (2 g, 0.0116 mol) and imidazole (1.58 g, 0.0232 mol) in DCM (40 mL) was added teri-butyl-chloro-dimethyl-silane (1.9 g, 0.0126 mol) at 0 °C. The reaction was warmed to room température and stirred for 4 hours. The reaction mixture was diluted with DCM (100 mL), washed with sat NaHCCh and brine, dried over Na2SÜ4, and concentrated under reduced pressure to get tert-butyl-(2-iodoethoxy)-dimethylsilane C44 (2.5 g, 68%). ’H NMR (400 MHz, Chlorofonn-7) δ 3.83 (t, J= 6.8 Hz, 2H), 3.20 (t, J = 6.8 Hz, 2H), 0.90 (s, 9H), 0.08 (s, 6H).

Step 2. Synthesis of l-[2-[tert-butyl(dimethyl)silyl]oxyethyl]pyrazole-4-carbaldehyde (S20) To a solution of lH-pyrazole-4-carbaldehyde C42 (20 g, 208.1 mmol) and K2CO3 (115 g, 832.1 mmol) in MeCN (200 mL) was added terZ-butyl-(2-iodoethoxy)-dimethyl-silane C44 (65 g, 227.1 mmol). The reaction was heated to 80 °C. The reaction was stirred for 5 hours. The reaction was cooled to 50 °C and stirred for 16 hours. The reaction mixture was allowed to reach ambient température, filtered, and solids were washed with MeCN (200 mL). The solids were discarded and the filtrate was concentrated. The residue was partitioned between EtOAc (400 mL) and water (400 mL). The organic layer was separated, washed with water (400 mL) and brine (400 mL), dried over MgSO4, filtered, and concentrated. Purification by silica gel chromatography (800 g column, 0-80% EtOAc in hexane) afforded the product. l-[2-[tertbutyl(dimethyl)silyl]oxyethyl]pyrazole-4-carbaldehyde S20 (46 g, 87%) as a pale yellow oïl. 'H NMR (300 MHz, Chloroformé) δ 9.86 (s, 1H), 7.98 (s, 2H), 4.25 (dd, J= 5.5, 4.5 Hz, 2H), 3.96 (dd, J= 5.5, 4.5 Hz, 2H), 0.83 (s, 9H), -0.06 (s, 6H). LCMS m/z 255.14 [M+H]⁺.

Préparation of S21 l-[3-[tert-butyl(dimethyl)silyl]bxy-2-[[tert-butyl(dimethyl)silyl] oxymethyl] -2-methylpropyl]pyrazole-4-carbaldehyde (S21)

O^X^OH ^{H Br} r- C45 C42 Biv N-, U | L· hnJ ₀ Si Si ------------------- / \ / \ C47

<^B \ ^Z —7—Si—CI -\^OH / \ C46 -* A^-0/^\/0^J\ Si Si / \ / \

168

Step 1. Synthesis of 2-(bromomethyl)-2-methyl-propane-l,3-diol (C46)

To a mixture of (3-methyloxetan-3-yl)methanol C45 (10 mL, 100.3 mmol) in THF (70 mL) at 0 °C was added hydrogen bromide (14 mL of 48% (w/w), 123.7 mmol). After stirring for 24 hours, the mixture was concentrated to a minimum volume, diluted in DCM/MeOH and the excess HBr was quenched with saturated sodium bicarbonate. The layers were split and the organic layer was dried with NazSCU, filtered, rinsed with methanol, and concentrated to yield 2(bromomethyl)-2-methyl-propane-l,3-diol C46 (13.6682 g, 74%). *H NMR (400 MHz, Methanol-^) δ 3.47 (d, J = 1.1 Hz, 6H), 0.96 (s, 3H).

Step 2. Synthesis of [2-(bromomethyl)-3-[tert-butyl(dimethyl)silyl]oxy-2-methyl-propoxy]tert-butyl-dimethyl-silane (C47)

To a mixture of 2-(bromomethyl)-2-methyl-propane-l,3-diol C46 (10 g, 54.09 mmol) in DCM (200 mL) was added imidazole (7.7 g, 113.1 mmol) followed by TBSC1 (17 g, 112.8 mmol). After 5 minutes, the mixture had precipitated a white crystalline solid. The mixture was filtered, rinsed with DCM, and concentrated. The mixture was diluted with heptane (25 mL) to further precipitate imidazole/imidazole HCl, filtered, and the solid was rinsed with additional heptane (10 mL). The mixture was concentrated, which precipitated additional solid. The mixture was diluted and concentrated twice more with heptane (50 mL) to afford [2(bromomethyl)-3-[tert-butyl(dimethyl)silyl]oxy-2-methyl-propoxy]-tert-butyl-dimethyl-silane C47 (22.246 g, 100%) ‘H NMR (400 MHz, Chloroformé) δ 3.44 (s, 4H), 3.40 (s, 2H), 0.94 (s, 3H), 0.89 (s, 18H), 0.04 (d, J= 1.2 Hz, 12H).

Step 3. Synthesis of l-[3-[tert-butyl(dimethyl)silyl]oxy-2-[[tertbutyl(dimethyl)silyl]oxymethyl]-2-methyl-propyl]pyrazole-4-carbaldehyde (S21)

To a vial was added lH-pyrazole-4-carbaldehyde C42 (2 g, 20.81 mmol), K2CO3 (4 g, 28.94 mmol), and [2-(bromomethyl)-3-[tert-butyl(dimethyl)silyl]oxy-2-methyl-propoxy]-tertbutyl-dimethyl-silane C47 (9.5 g, 23.08 mmol) in DMF (20 mL). The mixture was heated to 130 °C. After 3 hours the mixture was cooled to room température, diluted with water (100 mL) and heptane (100 mL). The layers were mixed, and the aqueous layer was washed with heptane (2 x 100 mL). The combined organic layers were washed with water (100 mL), brine (100 mL) and the organic layer was dried over NazSCM and concentrated. Purification by silica gel chromatography (Gradient: 0-60% EtOAc:Heptane) yielded the product l-[3-[tertbutyl(dimethyl)silyl] oxy-2-[ [tert-butyl(dimethyl)silyl] oxymethyl] -2-methyl-propyl]pyrazole-4carbaldehyde S21 (2.39 mg, 23%). ^rH NMR (400 MHz, Chloroformé) δ 9.85 (s, 1H), 7.98 7.91 (m, 2H), 4.12 (s, 2H), 3.43 - 3.29 (m, 4H), 0.91 (s, 18H), 0.84 (s, 3H), 0.05 (d, J= 0.6 Hz, 12H). LCMS m/z 427.31 [M+H]⁺.

169

Préparation of S22

2-[(2-hydroxy-l,l-dimethyl-ethyl)amino]pyrimidine-5-carbaldehyde (S22)

Step 1. Synthesis of ethyl 2-[(2-hydroxy-l, 1 -dimethyl-ethyl)amino]pyrimidine-5-carboxylate (C49)

To a stirred solution of ethyl 2-chloropyrimidine-5-carboxylate C48 (25 g, 0.1340 mol) in Ethanol (750 mL) was added 2-amino-2-methyl-propan-l-ol (14.333 g, 15.412 mL, 0.1608 mol) followed by DIPEA (34.637 g, 46.681 mL, 0.2680 mol) at room température. The reaction was stirred at 80 °C for 8 hours. The reaction was warmed to room température and concentrated under reduced pressure. Purification by silica gel chromatography (Eluent: 70% EtOAc in petroleum ether) afforded ethyl 2-[(2-hydroxy-l,l-dimethyl-ethyl)amino]pyrimidine5-carboxylate C49 (18 g, 55%). *H NMR (400 MHz, DMSO-î/ô) δ 8.70 (s, 2H), 7.39 (s, 1H), 4.86 (t, J= 6 Hz, 1H), 4.25 (q, J= 6.8 Hz, 2H), 3.52 (d, J= 6 Hz, 2H), 1.32 (s, 6H), 1.28 (t, J= 6.8 Hz, 3H). LCMS m/z 240.27 [M+H]⁺. '

Step 2. Synthesis of ethyl 2-[[2-[tert-butyl(dimethyl)silyl]oxy-l,l-dimethylethyl]amino]pyrimidine-5-carboxylate (C50)

To a stirred solution of ethyl 2-[(2-hydroxy-l,l-dimethyl-ethyl)amino]pyrimidine-5carboxylate C49 (10 g, 0.0410 mol) and tert-butyl-chloro-dimethyl-silane (9.2694 g, 0.0615 mol) in DCM (500 mL) was added imidazole (8.3735 g, 0.1230 mol) followed by DMAP (1.0018 g, 0.0082 mol) at room température and stirred for 16 hours. The reaction was concentrated under reduced pressure. The crude material was diluted with water (500 mL) and pentane (500 mL). The organic layer was separated, washed with water, dried over Na₂SO4, and 170 concentrated under reduced pressure to afford ethyl 2-[[2-[tert-butyl(dimethyl)silyl]oxy-l,ldimethyl-ethyl]amino]pyrimidine-5-carboxylate C50 (14.9 g, 100%). 'H NMR (400 MHz, DMSO-Ts) δ 8.70 (s, 2H), 7.46 (s, 1H), 4.25 (q, J= 7.2 Hz, 2H), 3.77 (s, 2H), 1.30 (s, 6H), 1.28 (t, J= 7.6 Hz, 3H), 0.82 (s, 9H), -0.06 (s, 6H). LCMS m/z 354.3 [M+H]⁺.

Step 3. Synthesis of [2-[[2-[tert-butyl(dimethyl)silyl] oxy-1,1-dimethylethyl]amino]pyrimidin-5-yl]methanol (C51)

To a stirred solution of ethyl 2-[[2-[tert-butyl(dimethyl)silyl]oxy-l,l-dimethylethyl]amino]pyrimidine-5-carboxylate C50 (15 g, 0.0411 mol) in THF (600 mL), was added DIBAL-H (1 M in Toluene) (205.50 mL of 1 M, 0.2055 mol) at -78 °C slowly under nitrogen. The reaction was stirred for 30 minutes at -78 °C and then warmed to room température and stirred for 4 hours. The reaction mixture was quenched with sat NH4CI (500 mL) at 0 °C and compound was extracted with EtOAc (2 x 500 mL). The organic layers were washed with 1 N HCl (100 mL), brine, dried over Na₂SÛ4, and concentrated under reduced pressure. Purification by silica gel chromatography (Eluent: 50% EtOAc in petroleum ether) afforded [2-[[2-[tertbutyl(dimethyl)silyl]oxy-l,l-dimethyl-ethyl]amino]pyrimidin-5-yl]methanol C51 (6 g, 46%). *H NMR (400 MHz, DMSO-7₆) δ 8.19 (s, 2H), 6.25 (s, 1H), 4.99 (t, J= 5.6 Hz, 1H), 4.27 (d, J= 5.6 Hz, 2H), 3.71 (s, 2H), 1.30 (s, 6H), 0.84 (s, 9H), -0.03 (s, 6H). LCMS m/z 312.23 [M+H]⁺.

Step 4. Synthesis of 2-[[2-[tert-butyl(dimethyl)silyl]oxy-1,1-dimethylethyl]amino]pyrimidine-5-carbaldehyde (C52)

To a stirred solution of [2-[[2-[tert-butyl(dimethyl)silyl]oxy-l,l-dimethylethyl]amino]pyrimidin-5-yl]methanol C51 (120 mg, 271.55 pmol) in DCM (10 mL) was added MnO₂ (851.98 mg, 0.0098 mol) at room température and stirred for 6 hours. The reaction was filtered through Celite® and washed with DCM (10 mL). The filtrâtes were concentrated under reduced pressure to provide 2-[[2-[tert-butyl(dimethyl)silyl]oxy-l,l-dimethylethyl]amino]pyrimidine-5-carbaldehyde C52 (90 mg, 99%). 'H NMR (400 MHz, DMSO-Ts) δ 9.71 (s, 1H), 8.71 (d, J= 11.6 Hz, 2H), 7.72 (s, 1H), 3.78 (s, 2H), 1.34 (s, 6H), 0.84 (s, 9H), 0.05 (s, 6H). LCMS m/z 310.22 [M+H]⁺.

Step 5. Synthesis of 2-[(2-hydroxy-l,l-dimethyl-ethyl)amino]pyrimidine-5-carbaldehyde (S22)

To a stirred solution of 2-[[2-[ter/-butyl(dimethyl)silyl]oxy-l,l-dimethylethyl]amino]pyrimidine-5-carbaldehyde C52 (2.9 g, 0.0087 mol) in THF (20 mL) was added TBAF (1 M in THF) (21.700 mL of 1 M, 0.0217 mol) at room température and stirred for 2 hours. The reaction was diluted with EtOAc (100 mL), washed with brine solution, dried over Na₂SO4, and concentrated under reduced pressure. The crude compound was washed with

171 pentane and dried to afford 2-[(2-hydroxy-l,l-dimethyl-ethyl)amino]pyrimidine-5-carbaldehyde S22 (1.47 g, 86%). Ή NMR (400 MHz, DMSO-îZ₆) δ 9.72 (s, 1H), 8.71 (d, J= 13.2 Hz, 2H), 7.64 (s, 1H), 4.87 (t, J= 6 Hz, 1H), 3.54 (d, J= 6 Hz, 2H), 1.33 (s, 6H). LCMS m/z 196.35 [M+H]⁺.

Préparation of S23 (3S)-3-aminobutanoic acid (S23)

S23 (3S)-3-aminobutanoic acid (S23) was obtained from commercial sources.

Préparation of S24

4-aminopentan-2-one hydrochloride (S24)

S24

4-aminopentan-2-one hydrochloride (S24) was obtained from commercial sources.

Préparation of S25 (4S)-4-aminopentan-2-one hydrochloride (S25)

Step 1. Synthesis of (3S)-3-(tert-butoxycarbonylamino)butanoic acid (C53)

To a solution of (35)-3-aminobutanoic acid S23 (100 g, 969.7 mmol) in dioxane (600 mL) was added aqueous NaOH solution (950 mL of 1 M, 950.0 mmol) over 15 minutes, followed by BoczO (300 g, 1.375 mol). The reaction mixture was stirred at room température for 12 hours. The reaction was partitioned with MTBE (1 L) and water (300 mL). The layers were separated, and the aqueous layer was extracted again with MTBE (500 mL). The aqueous layer

172 was then acidified with 1 N HCl until pH = 2 and extracted with DCM (3 x 600 mL). The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated in vacuo to yield (35)-3-(tert-butoxycarbonylamino)butanoic acid C53 (176 g, 89%) as a white solid. ‘H NMR (300 MHz, Chloroform-iZ) δ 4.92 (s, 1H), 4.04 (s, 1H), 2.56 (dd, J= 5.5, 2.9 Hz, 2H), 1.44 (s, 9H), 1.25 (d, J= 6.8 Hz, 3H).

Step 2. Synthesis of tert-butyl N-[(lS)-3-[methoxy(methyl)amino]-l-methyl-3-oxopropyl]carbamate (C54)

To a solution of (35)-3-(tert-butoxycarbonylamino)butanoic acid C53 (160 g, 787.3 mmol) in DCM (1.5 L) was added N-methoxymethanamine (Hydrochloride sait) (81 g, 830.4 mmol) followed by the addition of DIPEA (560 mL, 3.215 mol) over 10 minutes. The reaction mixture was cooled to 0 °C and T3P (600 g of 50% (w/w) in EtOAc, 942.9 mmol) was added over 45 minutes. After the addition, the cooling bath was removed and the reaction was stirred at room température for 1 hour. The reaction mixture was cooled to 10 °C and aqueous 1 N NaOH solution (700 mL) was added and the solution stirred for 15 minutes. The organic phase was separated, washed with aqueous saturated ammonium chloride solution (200 mL) and brine (200 mL), dried, filtered through a silica plug, and concentrated in vacuo to afford teri-butyl N-[(15)3-[methoxy(methyl)amino]-l-methyl-3-oxo-propyl]carbamate C54 (180 g, 93%) as a clear, colorless viscous oil. ’H NMR (300 MHz, Chloroform-i/) δ 5.30 (s, 1H), 4.06 (ddd, J= 14.3, 9.7, 6.0 Hz, 1H), 3.68 (s, 3H), 3.17 (s, 3H), 2.71 (dd, J= 15.6, 5.2 Hz, 1H), 2.54 (dd, J= 15.7, 5.7 Hz, 1H), 1.43 (s, 9H), 1.24 (d, J= 6.8 Hz, 3H).

Step 3. Synthesis of tert-butyl N-[(lS)-l-methyl-3-oxo-butyl]carbamate (C55)

To a solution of tert-butyl N-[(15)-3-[methoxy(methyl)amino]-l-methyl-3-oxopropyl]carbamate C54 (220 g, 893.2 mmol) in THF (4 L) at 0 °C was added iodo(methyl)magnesium (900 mL of 3 M, 2.700 mol) over 40 minutes. The resulting reaction mixture was stirred at 0 °C for 4 hours. The reaction was quenched with saturated ammonium chloride solution (2 L), followed by MTBE (1 L) and water (2 L). The mixture was stirred for 30 minutes and the organic layer was separated. The aqueous phase was extracted with MTBE (IL) and the combined organic layers were washed with saturated ammonium chloride solution (1 L), dried over MgSO4, filtered, and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-70% EtOAc in heptane) yielded the product tert-butyl N-[(15)-l-methyl-3-oxobutyl]carbamate C55 (115 g, 64%) as a white solid. *H NMR (300 MHz, Chloroform-ίΖ) δ 4.83 (s, 1H), 4.12 - 3.87 (m, 1H), 2.69 (dd, J= 16.5, 5.2 Hz, 1H), 2.63 - 2.47 (m, 1H), 2.15 (d, J= 2.3 Hz, 3H), 1.43 (d, J= 2.4 Hz, 9H), 1.20 (dd, J= 6.8, 2.4 Hz, 3H).

Step 4. Synthesis of (4S)-4-aminopentan-2-one (Hydrochloride sait) (S25)

173

To a solution of terAbutyl N-[(lS)-l-methyl-3-oxo-butyl]carbamate C55 (16.3 g, 80.18 mmol) in MeOH (30 mL) was added hydrogen chloride (50 mL of 4 M in dioxane, 200.0 mmol) over 3 minutes. The reaction was stirred at room température for 5 hours and then concentrated under reduced pressure. The residue was co-evaporated with EtOH (2x30 mL) and dried under vacuum to afford (45)-4-aminopentan-2-one (Hydrochloride sait) S25 (12 g, 98%) as apink viscous oil. *H NMR (300 MHz, Chloroform-7) δ 8.06 (s, 3H), 3.48 (d, J= 6.8 Hz, 1H), 2.88 (dd, J= 18.0, 5.8 Hz, 1H), 2.75 (dd, J= 18.0, 7.2 Hz, 1H), 2.13 (s, 3H), 1.17 (d, J= 6.6 Hz, 3H).

Préparation of S26 (Method A) (2S, 6S)-2-methyl-6-(l-methyltriazol-4-yl)piperidin-4-one (S26)

S25

L-Proline TEA

MgSO₄

Step 1. Synthesis of (2S)-2-methyl-6-(l-methyltriazol-4-yl)piperidin-4-one (C56)

To a mixture of (45)-4-aminopentan-2-one (Hydrochloride sait) S25 (12 g, 78.48 mmol) in EtOH (300 mL) was added l-methyltriazole-4-carbaldehyde S17 (9 g, 81.01 mmol), L-Proline (2 g, 17.37 mmol), magnésium sulfate (12 g, 99.69 mmol), and TEA (13 mL, 93.27 mmol). The reaction mixture was stirred at room température ovemight. The mixture was filtered and concentrated under reduced pressure. The crude residue was quenched with saturated sodium bicarbonate solution (150 mL) and extracted with DCM (3 x 100 mL). The combined organic layers were washed with brine (50 mL), dried over magnésium sulfate, filtered, and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-60% of 20% MeOH/DCM in DCM) yielded the product (25)-2-methyl-6-(l-methyltriazol-4-yl)piperidin-4-one C56 (6.7 g, 44%) as a 5:1 cis to trans ratio. Additionally, the e.r. at the stereocenter from S25 was eroded to -85%.

NMR for the major (CIS) stereoisomer in C56: 'H NMR (300 MHz, Chloroform-7) δ 7.47 (s, 1H), 4.26 (dd, J= 10.3, 4.9 Hz, 1H), 4.11 (s, 3H), 3.17 (dqd, J= 12.2, 6.2, 3.0 Hz, 1H), 2.73 - 2.56 (m, 2H), 2.47 (ddd, J= 14.2, 3.0, 1.6 Hz, 1H), 2.21 (dd, J= 14.2, 11.7 Hz, 2H), 1.28 (d, 7=6.2 Hz, 3H).

NMR Rationalization of Stereoisomer Assignments in C56: Note that major component in C56 was assigned as the cis stereoisomer using NMR coupling constant data for the peak at 4.26 ppm (C5-methylene proton). The triazole at C6 is assumed to occupy an équatorial position 174 in the lowest energy conformation. The coupling between the axial CH at C4 and one of the CH protons at C5 (J = 10.3 Hz) indicates a 180° relationship as defined by the Karplus équation. The minor trans product was removed in the subséquent re-crystallization step to afford S26.

Step 2. Synthesis of (2S,6S)-2-methyl-6-(l-methyltriazol-4-yl)piperidin-4-one (S26)

A solution of (2S)-2-methyl-6-(l-methyltriazol-4-yl)piperidin-4-one C56 (6.7 g) as a 5:1 cis to trans ratio in MTBE (100 mL) was heated to reflux for 30 minutes. Ethanol was added slowly until ail solids dissolved (20 mL). The solution was refluxed for 30 minutes and allowed to slowly cool ovemight. A solid crystalized out which was diluted with MTBE (30 mL), filtered, and dried under vacuum to afford the (2S,65)-2-methyl-6-(l-methyltriazol-4yl)piperidin-4-one S26 (3.2 g, 48%) as a white solid with an enantiomeric ratio of >85% which was carried through to ail further compounds which utilized S26 as a starting material unless otherwise noted (excluding examples which were subjected to SFC purification). 'H NMR (300 MHz, Chloroform-<7) δ 7.45 (s, 1H), 4.23 (dd, J= 10.3, 4.9 Hz, 1H), 4.09 (s, 3H), 3.14 (ddp, J= 12.2,6.1,3.1 Hz, 1H), 2.71 - 2.52 (m, 2H), 2.44 (ddd, J= 14.1, 3.0, 1.5 Hz, 1H), 2.27 - 2.00 (m, 2H), 1.26 (d, J =6.2 Hz, 3H).

Alternative Préparation of S26 (Method B) ((2S, 6S)-2-methyl-6-(l-methyltriazol-4-yl)piperidin-4-one (S26)

Mg(OtBu)₂

C108

Step 1. Synthesis ofbis[(3-tert-butoxy-3-oxo-propanoyl)oxy] magnesium(C109)

175

A solution of 3-tert-butoxy-3-oxo-propanoic acid C108 (321.51 g, 1.907 mol) in THF (2 L) was cooled to 5 °C in an ice-bath and Mg(OEt)2 (111.33 g, 953.5 mmol) was added. The reaction was stirred for 30 minutes at 0 °C, removed from the cooling bath and stirred at room température ovemight. The reaction was fïltered over a plug of Celite® and the plug was washed with additional THF. The clear, colorless filtrate was evaporated in vacuo to afford a mushy solid. The solid was triturated with 1 L of diethyl ether and fïltered. The filter-cake was washed with Et₂O and dried in vacuo. The filtrate was evaporated in vacuo again and was then triturated with a small volume of Et2Ü and fïltered to afford a second crop of the product. The crops were combined and dried in vacuo to afford bis[(3-tert-butoxy-3-oxo-propanoyl)oxy]magnesium C109 (294.49 g, 90%) as a white solid. Ή NMR (300 MHz, Methanol-ck) δ 4.92 (s, 4H), 1.48 (s, 18H) ppm.

Step 2. Synthesis of tert-butyl (5S)-5-(tert-butoxycarbonylamino)-3-oxo-hexanoate (Clll) To a solution of (3S)-3-(tert-butoxycarbonylamino)butanoic acid C110 (170.15 g, 837.2 mmol) in THF (1.5 L) was added CDI (149.8 g, 923.8 mmol). The milky suspension cleared over the next few minutes. Gas évolution was observed. The reaction was stirred for 3 hours at room température. Bis[(3-tert-butoxy-3-oxo-propanoyl)oxy]magnesium C109 (172.19 g, 502.6 mmol) was added. Another milky suspension was formed that cleared after stirring for 30 minutes. The reaction was stirred for 48 hours. The reaction was poured into 1.5 L of 1 N HCl and extracted with MTBE (IL). The pH was confirmed to be approximately pH 3. The extract was washed with saturated aqueous NaHCCh, separated, dried with MgSCL, fïltered, and evaporated in vacuo to afford tert-butyl (5S)-5-(tert-butoxycarbonylamino)-3-oxo-hexanoate Clll (248.5 g, 98.5%). ‘H NMR (300 MHz, Chloroform-ίΖ) δ 4.90 (d, J = 18.1 Hz, 1H), 4.04 (dt, J = 13.8, 6.6 Hz, 1H), 3.47 - 3.22 (m, 2H), 2.76 (qd, J = 17.0, 5.7 Hz, 2H), 1.48 (s, 9H), 1.44 (s, 9H), 1.23 (d, J = 6.8 Hz, 3H) ppm.

Step 3. Synthesis of tert-butyl (2S,3R,6S)-6-methyl-2-(l-methyltriazol-4-yl)-4-oxopiperidine-3-carboxylate (C112)

To a solution of tert-butyl (5S)-5-(tert-butoxycarbonylamino)-3-oxo-hexanoate Clll (248.5 g, 824.5 mmol) in DCM (1.5 L) was added TFA (240 mL, 3.115 mol) and the reaction was stirred ovemight. The reaction was evaporated in vacuo at 25 °C. The solid that remained was triturated with 500 mL of pentane and fïltered. The filter-cake was washed with pentane and most of the solvent was pulled off of the filter-cake. The cake was transferred back to the reaction flask and dissolved in 1 L of DCM.

l-methyltriazole-4-carbaldehyde S17 (120.7 g, 1.086 mol) was added. The reaction was stirred at room température ovemight. Brine (100 mL) was added and 6N NaOH was added until 176 the aqueous layer remained alkaline when the tunnel was shaken. The organic layer was isolated and the aqueous layer was extracted with DCM (1 L). The organic layers were combined, dried with MgSCU, and filtered over a plug of silica gel. The plug was eluted with 10% MeOH in EtOAc. The filtrate was evaporated in vacuo to afford a solid that was triturated with MTBE (500 mL) and filtered. The filter-cake was washed with MTBE and dried in vacuo to give a crop of product. The mother liquor from the trituration was concentrated. The solid that precipitated was filtered to provide a second crop of the product. The crops were combined to give (2S,3R,6S)-6-methyl-2-(l-methyltriazol-4-yl)-4-oxo- piperidine-3-carboxylate C112 (105.45 g, 43%). *H NMR (300 MHz, Chloroform-ίΖ) δ 7.48 (s, 1H), 4.52 (d, J = 11.0 Hz, 1H), 4.09 (s, 3H), 3.61 (dd, J = 11.0, 1.0 Hz, 1H), 3.21 (ddd,J=11.7, 6.1,2.9 Hz, 1H), 2.55 (dd, J = 13.7, 2.9 Hz, 1H), 2.37 - 2.13 (m, 1H), 1.98 (s, 1H), 1.39 (s, 9H), 1.29 (d, J = 6.3 Hz, 3H) ppm.

Step 4. Synthesis of (2S,6S)-2-methyl-6-(l-methyltriazol-4-yl)piperidin-4-one (S26) To a solution of tert-butyl (2S,3R,6S)-6-methyl-2-(l-methyltriazol-4-yl)-4-oxopiperidine-3-carboxylate C112 (70.59 g, 239.8 mmol) in DCM (750 mL) was added MsOH (62 mL, 955.4 mmol) and the reaction was heated to reflux for 6 hours. The reaction was cooled and poured into a separatory funnel. Brine (approx. 100 mL) was added. 6N NaOH was added until the aqueous layer remained alkaline after shaking. The organic layer was separated and the aqueous was extracted with DCM (2 x 500 mL). The organic layers were combined, dried with MgSCfi, filtered, and evaporated in vacuo to afford (2S,6S)-2-methyl-6-(l-methyltriazol-4yl)piperidin-4-one S26 (43.74 g, 94%). Ή NMR (300 MHz, Chloroform-J) δ 7.46 (s, 1H), 4.20 (dd, J = 10.1, 5.1 Hz, 1H), 4.06 (s, 3H), 3.11 (dqd, J = 12.3, 6.2, 3.0 Hz, 1H), 2.73 - 2.48 (m, 2H), 2.40 (ddd, J = 14.1, 3.0, 1.5 Hz, 1H), 2.25 - 2.00 (m, 2H), 1.23 (d, J = 6.2 Hz, 3H) ppm.

Préparation of S27-S29

Intermediates S27-S29 (see Table 1) were prepared in a single step from intermediate S25 using the appropriate aldéhyde and the method described for intermediate S26 (Method A). Aldéhydes were prepared by methods described above or obtained from commercial sources. As for intermediate S26 (prepared by method A), partial stereochemical érosion of the enantiomerically pure starting material (45)-4-aminopentan-2-one (Hydrochloride sait) S25 was observed in step 1 leading to unseparated mixtures of stereoisomers being generated in Step 1. In each case, the cis-product was the major isomer. This mixture is represented by use of wavy bonds. Any modifications to methods are noted in Table 1 and accompanying footnotes.

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Table 1. Method of préparation, structure and physicochemical data for intermediates S27-S29

Product	Aldéhyde Reagent	Method	¹II NMR
O Ν.γΧ S27 /	O A <T^N N 1	Préparation ofS26^1,2·^{1 2 3}·⁴	Ή NMR (300 MHz, Chloroform-ίΖ) δ 7.45 (s, 1H), 7.34 (s, 1H), 3.97 (dd, J= 11.5,3.4 Hz, 1H), 3.87 (s, 3H), 3.07 (dqd, J= 12.3, 6.2, 2.9 Hz, 1H), 2.53 (ddd, J= 14.0, 3.4, 2.0 Hz, 1H), 2.47 2.35 (m, 2H), 2.14 (ddd, J= 14.1, 11.6, 1.1 Hz, 1H), 1.23 (d, .7=6.2 Hz, 3H).
O \ II H Isr S28/	O y-n o N 1	Préparation ofS26²·⁴·⁵	Ή NMR (300 MHz, Chloroform-ίΖ) δ 7.40 (s, 1H), 6.78 (s, 1H), 4.02 (dd, J= 9.6, 5.5 Hz, 1H), 3.67 (s, 3H), 3.12 (dtt, J= 12.1, 6.1, 3.0 Hz, 1H), 2.66 - 2.58 (m, 2H), 2.47 - 2.38 (m, 1H), 2.17 (dd, J= 14.1, 11.6 Hz, 1H), 1.26 (d, .7=6.3 Hz, 3H).
S29 O N? J K	O O N O=S-° S18	Préparation ofS26²·⁴·⁵	Ή NMR (300 MHz, Chloroform-iZ) δ 7.58 (s, 1H), 7.53 (s, 1H), 4.60 (t, J = 6.3 Hz, 2H), 4.00 (dd, J= 11.6, 3.3 Hz, 1H), 3.65 (t, J= 6.2 Hz, 2H), 3.10 (dqd, J= 12.1, 6.0, 2.9 Hz, 1H), 2.58 - 2.51 (m, 4H), 2.48 - 2.37 (m, 2H), 2.17 (dd, J= 14.1, 11.6 Hz, 1H), 1.26 (d, .7=6.1 Hz, 3H).
O N J H S30 Hhr	N-, ^ηνΎ^,ο C42	Préparation ofS26⁴·⁵	*H NMR (300 MHz, Chloroformé δ 7.60 (s, 2H), 4.05 (dd, J= 11.6, 3.4 Hz, 1H), 3.10 (ddd, J= 11.8, 6.0, 2.9 Hz, 1H), 2.63 - 2.34 (m, 3H), 2.18 (dd, J = 14.0, 11.5 Hz, 1H), 1.26 (d, J = 6.2 Hz, 3H).

1. Reaction was stirred over the weekend (step 1)

2. The crude residue was diluted with water and saturated sodium bicarbonate solution and extracted with DCM (5x) through a phase separator. (step 1)

3. Purification by silica gel chromatography (Gradient: 0-50% of 20% MeOH/DCM in DCM) yielded the product. (step 1)

4. The minor isomer was purged via chromatography and step 2 was not performed.

5. Purification by silica gel chromatography (Gradient: 0-100% of 20% MeOH/DCM in DCM) yielded the product. (step 1)

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Compound 1 (2'S,6'S, 7S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran7,4'-piperidine] (1)

To a solution of (25,65)-2-methyl-6-(l-methyl triazol-4-yl)piperidin-4-one S26 (1380 mg, 7.11 mmol, S26 was prepared by Method A) in DCM (30 mL) was added 2-(5-chloro-3thienyl)ethanol S2 (1100 pL, 8.894 mmol) followed by MsOH (3 mL, 46.23 mmol). The reaction was heated to reflux for 90 minutes at which time it was cooled to room température and quenched with 2 N NaOH until the pH reached 14. The mixture was diluted with DCM (20 mL) and the organic layer separated, washed with brine (30 mL), dried over MgSCU, and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-25% of 20% MeOH/DCM in DCM) yielded the product (2'5,6'5,75)-2-chloro-2^l-methyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] 1 (1162 mg, 48%) as a pale yellow oil in a >8:1 ratio. The minor isomer observed is inferred to be the enantiomer of compound 1, since S26 prepared by method A contains minor quantities of the other cis enantiomer. Note that relative stereôchemistry in compound 1 was assigned through NOE NMR studies. *H NMR (400 MHz, Chloroformé) δ 7.42 (s, 1H), 6.58 (s, 1H), 4.41 (dd, J= 11.8, 2.6 Hz, 1H), 4.06 (s, 3H), 4.02 3.86 (m, 2H), 3.30 (ddt, J= 12.7, 6.3, 3.2 Hz, 1H), 2.70 - 2.49 (m, 2H), 2.35 (dt, J= 13.6, 2.6 Hz, 1H), 2.06 (dt, J=13.7, 2.5 Hz, 1H), 1.79 (dd,J=13.6, 11.8 Hz, 1H), 1.42 (dd,J=13.7,11.3 Hz, 1H), 1.31-1.19 (m, 1H), 1.12 (d, J= 6.4 Hz, 3H). LCMS m/z 339.0 [M+H]⁺.

Alternative Préparation of Compound 1 (HCl sait) (2 'S, 6'S, 7S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] hydrochloride sait (1) (25,65)-2-methyl-6-(l-methyltriazol-4-yl)piperidin-4-one S26 (205 mg, 1.055 mmol) in DCM (5 mL) was added 2-(5-chloro-3-thienyl)ethanol S2 (150 pL, 1.213 mmol) followed by MsOH (300 pL, 4.623 mmol). The mixture was heated to reflux for 10 minutes at which time it was cooled to room température and quenched with 2 N NaOH until the pH reached 14. The mixture was diluted with DCM (5 mL) and the organic layer was separated and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-25% of 20% MeOH/DCM in

179

DCM) yielded product which was immediately dissolved in minimal DCM and treated with HCl (100 pL of 4 M in dioxane, 0.4000 mmol). The mixture was concentrated in vacuo and the residue was azeotroped with DCM (5 mL) and dried to yield (2'5,6'5,75)-2-chloro-2'-methyl-6'(l-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (Hydrochloride sait) 1 (171.6 mg, 43%) as a pale yellow solid. *H NMR (300 MHz, DMSO-t/ό) δ 9.46 (s, 1H), 9.24 (d, J= 8.3 Hz, 1H), 8.29 (s, 1H), 6.95 (s, 1H), 4.67 (t, J= 11.1 Hz, 1H), 4.09 (s, 3H), 3.95 (t, J= 5.4 Hz, 2H), 3.72 (s, 1H), 2.61 (t, J= 5.3 Hz, 2H), 2.46 - 2.32 (m, 2H), 2.25 (d, J= 15.1 Hz, 1H), 2.01 - 1.86 (m, 1H), 1.29 (d, J= 6.5 Hz, 3H). LCMS m/z 339.0 [M+H]⁺

Compound2 (2'S,6'S, 7S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4 ’-piperidine] (2)

To a solution of (2S,65)-2-methyl-6-(l-methyltriazol-4-yl)piperidin-4-one S26 (250 mg, 1.287 mmol) and 2-[5-(trifluoromethyl)-3-thienyl]éthanol S3 (350 mg, 1.748 mmol) in DCM (5 mL) was added MsOH (500 pL, 7.705 mmol) and the reaction was heated to 40 °C. After 16 hours, additional MsOH (200 pL, 3.082 mmol) was added and the reaction was continued heating ovemight. The mixture was diluted with water (4 mL) and DCM (5 mL) and quenched with aqueous NaOH (2 mL of 6 M, 12.00 mmol). The mixture was separated, extracted with DCM (2x5 mL), passed over a phase separator, and the organics concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-10% MeOH in DCM) yielded (2'5,6'5,75)-2'-methyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] 2 (445 mg, 93%) as a white solid. Note that relative stéréochemistry in Compound 2 was assigned through NOE NMR studies. *H NMR (300 MHz, Chloroform-<7) δ 7.46 (s, 1H), 7.14 (s, 1H), 4.47 (d, J= 11.6 Hz, 1H), 4.08 (d, J= 3.3 Hz, 3H), 4.00 (s, 2H), 3.36 (s, 1H), 2.72 (d, J= 5.6 Hz, 2H), 2.41 (d, J= 14.2 Hz, 1H), 2.12 (d, J= 13.7 Hz, 1H), 1.86 (t, J= 12.7 Hz, 1H), 1.49 (d, J= 12.8 Hz, 1H), 1.15 (d,J= 6.3 Hz, 3H). LCMS m/z 373.07 [M+H] ⁺

Compounds 3-16

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Compounds 3-16 (see Table 2) were prepared from a single Oxa-Pictet Spengler step with isolated piperidones (S26, S29, or C56) and the relevant thiophene ethanols as described for compounds 1 and 2. Thiophene ethanols and piperidone were prepared by methods described above or obtained from commercial sources. In examples where S26 was used, S26 was prepared 5 by Method A, therefore the piperidone used may contain minor amounts of the other cis-isomer.

Any modifications to methods are noted in Table 2 and accompanying footnotes.

Table 2. Method of préparation, structure and physicochemical data for Compounds 3-16.

Product	Piperidone and Thiophene éthanol	Method	‘HNMR; LCMS m/z [Μ+ΙΙΓ
Compound 3 ⁿ:j) h /	S26; VJ HO S14	Compound 11.2,3	‘H NMR (300 MHz, Methanolé) δ 8.03 (s, 1H), 6.51 (s, 1H), 4.92 - 4.85 (m, 1H), 4.12 (s, 3H), 3.99 (t, J= 5.4 Hz, 2H), 3.79 (s, 1H), 2.64 (t, J= 5.5 Hz, 2H), 2.50 (d, J= 15.1 Hz, 1H), 2.42 (d, J= 1.1 Hz, 3H), 2.37 - 2.27 (m, 2H), 1.88 (dd, J= 14.8, 12.3 Hz, 1H), 1.39 (d, J= 6.6 Hz, 3H). LCMS m/z 319.2 [M+H]⁺
Compound 4 N'J) H /	S26; ^X^^ HO S5	Compound 12.3	Ή NMR (400 MHz, DMSOé) δ 9.36 (d, J = 23.7 Hz, 2H), 8.31 (s, 1H), 6.55 (s, 1H), 4.63 (t, J =11.2 Hz, 1H), 4.08 (s, 3H), 3.93 (t, J = 5.3 Hz, 2H), 3.57 (s, 1H), 2.80 - 2.66 (m, 4H), 2.43 (d, J = 13.4 Hz, 1H), 2.20 (d, J = 14.1 Hz, 1H), 2.09 - 1.96 (m, 2H), 1.30 (d, J = 6.4 Hz, 3H), 1.21 (t, J = 7.5 Hz, 3H). LCMS m/z 333.2 [M+H]⁺
Compound 5 oOO^-^ î? 0	S29; ^X^ HO S5	Compound 21,3,4,5	Ή NMR (300 MHz, DMSOé) δ 9.24 - 8.96 (m, 2H), 8.06 (s, 1H), 7.79 (s, 1H), 6.57 (s, 1H), 4.54 (t, J = 6.8 Hz, 2H), 4.44 (t, J= 10.9 Hz, 1H), 3.91 (t, J= 5.3 Hz, 2H), 3.67 (t, J= 6.8 Hz, 2H), 3.51 (s, 1H), 2.85 (s, 3H), 2.81 - 2.67 (m, 4H), 2.41 (d, J= 13.8 Hz, 1H), 2.13 - 1.98 (m, 3H), 1.29 (d, J =6.4 Hz, 3H), 1.22 (t, J= 7.5 Hz, 3H). LCMS m/z ' 424.18 [M+H]⁺

181

Product	Piperidone and Thiophene éthanol	Method	’HNMR; LCMS m/z [M+Hf
Compound 6 N' J H /	S26; ^<vj^s HO S4	Compound l²’³	Ή NMR (400 MHz, DMSO-7₆) δ 9.58 - 9.19 (m, 2H), 8.30 (s, 1H), 6.59 (s, 1H), 4.66 (q, J = 8.8 Hz, 1H), 4.09 (s, 3H), 3.92 (t, J = 5.4 Hz, 2H), 3.58 (s, 1H), 2.76 (q, J = 7.5 Hz, 2H), 2.59 (t, J = 5.4 Hz, 2H), 2.36 (d, J = 8.0 Hz, 2H), 2.20 (d, J = 13.3 Hz, 1H), 1.95 (dd, J =14.4, 12.2 Hz, 1H), 1.30 (d, J = 6.4 Hz, 3H), 1.21 (t, J = 7.5 Hz, 3H). LCMS m/z 333.2 [M+H]⁺
Compound 7 (OlV^ci H'Jj H hr /	C56; Cl HO S2	Compound 16.7.8.9	Ή NMR (300 MHz, Chloroform-7) δ 7.43 (s, 1H), 6.57 (s, 1H), 4.62 (dd, J= 11.6, 2.6 Hz, 1H), 4.06 (s, 3H), 3.94 (t, J= 5.5 Hz, 2H), 3.48 (s, 1H), 2.60 (td, J= 5.5, 2.6 Hz, 2H), 2.33 (dt, J= 13.5, 2.6 Hz, 1H), 2.08 (dt, J = 14.4, 2.2 Hz, 1H), 1.93 (dd, J= 14.5,6.1 Hz, 1H), 1.84 (dd,7=13.5, 11.7 Hz, 1H), 1.46 (d, 7=7.1 Hz, 3H). LCMS m/z 339.09 [M+H]⁺
Compound 8 OH ^f <r ή 1Ή /	S26; J.F Λ ^S HO S6	Compound J2.3	Ή NMR (400 MHz, DMSO-J₆) δ 9.41 (d, J = 26.0 Hz, 2H), 8.33 (s, 1H), 7.43 (s, 1H), 4.71 -4.57 (m, 1H), 4.09 (s, 3H), 4.00 (t, J = 5.4 Hz, 2H), 3.59 (s, 1H), 2.91 (t, J = 5.3 Hz, 2H), 2.58 (d, J = 13.9 Hz, 1H), 2.28 (d, J = 14.3 Hz, 1H), 2.14 (dt, J = 25.5, 14.1 Hz, 2H), 1.31 (d, J = 6.5 Hz, 3H). LCMS m/z 373.16 [M+H]⁺
Compound 9 Pi ^F j_Ajr ί N-' 0	S29; Λ ^S HO S6	Compound p,3.4,5	Ή NMR (300 MHz, Methanol-70 δ 7.97 (s, 1H), 7.75 (s, 1H), 7.47 (s, 1H), 4.74 - 4.68 (m, 1H), 4.65 (t, 7= 6.4 Hz, 2H), 4.04 (t, 7= 5.4 Hz, 2H), 3.78 (s, 1H), 3.70 (t, 7= 6.4 Hz, 2H), 2.93 (t, 7= 5.5 Hz, 2H), 2.84 (s, 3H), 2.40 - 2.31 (m, 2H), 2.21 (d,7= 14.5 Hz, 1H), 2.02 - 1.88 (m, 1H), 1.39 (d, 7= 6.6 Hz, 3H). LCMS m/z 464.1 [M+H]⁺
Compound 10	S26;	Compound l²’³	‘H NMR (400 MHz, DMSO-7₆) δ 9.40 (s, 2H), 8.32 (s, 1H), 6.53 (s, 1H), 4.63 (s, 1H), 4.08 (s, 3H), 3.96

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Product	Piperidone and Thiophene éthanol	Method	‘H NMR; LCMS m/z [M+ilf
~ \ V\ o '—{ Z	ÇÎ _____(* HO S12		(dd, J = 11.4, 4.6 Hz, 1 H), 3.643.48 (m, 2H), 2.94 (d, J = 10.9 Hz, 1H), 2.77 (q, J = 7.5 Hz, 2H), 2.39 (d, J = 13.2 Hz, 1H), 2.28 (d, J = 14.3 Hz, 1H), 2.10 (t, J = 13.2 Hz, 1H), 1.95 (d, J = 14.1 Hz, 1H), 1.30 (d, J = 6.5 Hz, 3H), 1.22 (t, J = 7.5 Hz, 3H), 1.15 (d, J = 6.8 Hz, 3H). LCMS m/z 347.24 [M+H]⁺
Compound 11 * rV^s\ / o *JLy l\r /	S26; Çl —-Λ.* HO S13	Compound l²·³	‘H NMR (400 MHz, DMSO-î/₆) δ 9.36 (d, J = 22.0 Hz, 2H), 8.31 (d, J = 1.9 Hz, 1H), 6.52 (s, 1H), 4.64 (t, J = 11.3 Hz, 1H), 4.08 (s, 3H), 3.96 (dd, J= 11.5, 4.6 Hz, 1H), 3.55 (s, 3H), 3.00 - 2.89 (m, 1H), 2.76 (q, J = 7.5 Hz, 2H), 2.12 (d, J = 15.6 Hz, 1H), 2.05 - 1.89 (m, 1H), 1.30 (d, J = 6.5 Hz, 3H), 1.22 (t, J = 7.5 Hz, 3H), 1.16 (t, J = 6.8 Hz, 3H). LCMS m/z 347.24 [M+H]⁺
Compound 12 * X T Jhci N'J H IsH /	S26; Cl / ^Sy* HO S10	Compound ;2,3	Ή NMR (400 MHz, DMSO-îZ₆) δ 9.46 (d, J = 26.5 Hz, 2H), 8.33 (s, 1H), 6.81 (s, 1H), 4.63 (t, J = 11.0 Hz, 1H), 4.08 (s, 3H), 3.99 (dd, J = 11.5, 4.5 Hz, 1H), 3.57 (dd, J = 11.5, 6.8 Hz, 2H), 2.98 (dd, J = 10.6, 5.5 Hz, 1H), 2.10 (dt, J = 40.8, 13.6 Hz, 3H), 1.30 (d, J = 6.4 Hz, 3H), 1.18 (d, J = 6.9 Hz, 3H). LCMS m/z 353.16 [M+H]⁺
Compound 13 * I T J^^ci N fi - N-^ !	S26; /\/^cl / ^S HO SU	Compound l²’³	‘H NMR (400 MHz, DMSO-J₆) δ 9.43 (s, 2H), 8.32 (s, 1H), 6.82 (d, J = 1.6 Hz, 1H), 4.73 - 4.54 (m, 1H), 4.09 (s, 3H), 3.99 (dd, J = 11.6, 4.6 Hz, 1H), 3.57 (dd, J = 11.6, 6.8 Hz, 2H), 3.03 - 2.93 (m, 1H), 2.45 - 2.36 (m, 1H), 2.31 (d, J = 14.6 Hz, 1H), 2.07 (dt, J = 52.7, 14.0 Hz, 2H), 1.30 (d, J = 6.4 Hz, 3H), 1.17 (t, J = 6.9 Hz, 3H). [1] LCMS m/z 353.16 [M+H]⁺
Compound 14	S26;	Compound l²'³	’H NMR (400 MHz, DMSO-J₆) δ 9.41 (d, J = 25.4 Hz, 2H), 8.33 (s, 1H), 7.43 (s, 1H), 4.71 - 4.60 (m,

183

Product	Piperidone and Thiophene éthanol	Method	'H NMR; LCMS m/z [M+Hf
/^F _o ^r JCm-^fΧ^χ*/Χ^^ί/ p N 'Jj B !	F L-f Pj ^f HC) S7		1H), 4.08 (s, 3H), 4.06 - 3.97 (m, 1H), 3.62 (dd, J = 11.7, 6.8 Hz, 2H), 3.14 (q, J = 6.5 Hz, 1H), 2.61 (d, J = 13.6 Hz, 1H), 2.40 - 1.96 (m, 3H), 1.31 (d, J = 6.4 Hz, 3H), 1.24 (t, J = 6.9 Hz, 3H). LCMS m/z 387.16 [M+H]⁺
Compound 15 n I^^CIϋχ* Νχ'ΡΛ, N'J H /	S26; ^.Cl ^S HO S9	Compound J2.3	’H NMR (400 MHz, DMSO-7₆) δ 9.49 (d, J = 53.8 Hz, 2H), 8.34 (s, 1H), 6.78 (s, 1H), 4.64 (t, J = 11.3 Hz, 1H), 4.09 (s, 3H), 3.65 (s, 2H), 3.59 (s, 1H), 2.48 (s, 1H), 2.25 (d, J = 14.2 Hz, lH),2.10(d, J= 11.2 Hz, 2H), 1.31 (d, J = 6.5 Hz, 3H), 1.24 (d, J = 6.6 Hz, 6H). LCMS m/z 367.16 [M+H]⁺
Compound 16 z^Fx^x*/^^·'··^/ p Nw'-P./P N J M /	S26; F Lf ,CC^F HO^Z S8	Compound l^{1 2}·³	‘H NMR (400 MHz, DMSO-J₆) δ 9.51 (d, J = 23.4 Hz, 2H), 8.35 (s, 1H), 7.39 (s, 1H), 4.67 (t, J =11.1 Hz, 1H), 4.09 (s, 3H), 3.70 (s, 2H), 3.62 (s, 1H), 2.60 (d, J= 13.9 Hz, 1H), 2.37 - 2.27 (m, 1H), 2.27 - 2.06 (m, 2H), 1.31 (t, J = 6.5 Hz, 9H). LCMS m/z 401.16 [M+H]⁺
S31 oOn^^{cl u}>çs ^ΗΝ3'Ή	S30; Cl HO S2	Compound j8J0	*H NMR (300 MHz, Chloroform-J) δ 7.57 (s, 2H), 6.58 (s, 1H), 4.22 (dd, 7=11.6, 2.5 Hz, 1H), 3.95 (t, 7=5.5 Hz, 2H), 3.38 - 3.13 (m, 1H), 2.61 (t, 7= 5.4 Hz, 2H), 2.40 - 2.16 (m, 1H), 2.16- 1.99 (m, 1H), 1.71 (dd,7= 13.6, 11.6 Hz, 1H), 1.41 (dd,7= 13.7, 11.3 Hz, 1H), 1.13 (d,7=6.4 Hz, 3H). LCMS m/z 324.02 [M+l]⁺

1. The reaction was stirred for 30 minutes.

2. After completion, the mixture was concentrated and diluted in MeOH. No further workup was done.

184

3. Purification by reversed-phase HPLC (Method: C18 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 5 mM HCl) yielded the product as the HCl sait.

4. The organic layer was collected through a phase separator and dried under nitrogen.

5. After purification ,the product was brought up in 0.6 mL of water, ffozen, and lyophilized ovemight to afford.

6. The reaction was stirred ovemight.

7. Once the reaction had gone to completion the organic layer was separated, dried over Na2SO4, filtered, and concentrated.

8. Purification by silica gel chromatography (Gradient: 0-20% MeOH in DCM) yielded the product.

9. Reaction was run with C56 which was enriched as a mixture of two isomers from purification of S26. Compound 7 was isolated as the minor product of the Pictet Spangler reaction as a single diastereomer. As described for Method A of S26, epimerization of the S25 stereocenter was observed, which provided this compound as a mixture of enantiomers.

10. After 50 minutes, the reaction was quenched with saturated NaHCCb solution and extracted with DCM (6x). The combined organic layer was dried over Na2SÜ4, filtered, and concentrated.

Compound 17

[(2'S,6'S, 7S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-3-yl]methanol (17)

Step 1. Synthesis of[(2S,6S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-3-yl]methyl acetate (C57)

To a mixture of (2S,6S)-2-methyl-6-(l-methyltriazol-4-yl)piperidin-4-one S26 (10 mg, 0.05148 mmol) and [4-(2-hydroxyethyl)-2-(trifluoromethyl)-3-thienyl]methyl acetate S16 (18 mg, 0.06710 mmol) in DCM (500 pL) was added MsOH (30 pL, 0.4623 mmol) and the mixture was heated to 40 °C. After stirring for 4 hours, the reaction was quenched with saturated

185

NaHCCh solution, the layers were separated and the mixture was concentrated to dryness to give crudë C57.

Step 2. Synthesis of [(2S,6S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5dihydrothieno[2,3-c]pyran- 7,4 '-piperidine]-3-yl]methanol (17)

The crude material C57 was diluted with MeOH (2 mL) and to the mixture was added NaOH (20 pL of 6 M, 0.1200 mmol). The reaction was stirred for 5 minutes. The mixture was concentrated, re-diluted in DCM and washed with brine. The organic layer was passed over a phase separator, and concentrated. Silica gel chromatography (Gradient: 0-20% MeOH-DCM) yielded 17 as the parent.

(parent) from the deprotection was diluted with diethyl ether (1 mL) and HCl (13 pL of 4 M in dioxane, 0.05200 mmol) was added which immediately precipitated a white solid. The mixture was concentrated, and azeotroped three times with diethyl ether to yield [(2S,6S)-2'methyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-3-yl]methanol 17 (Hydrochloride sait) (10.9 mg, 45%). *H NMR (300 MHz, DMSOd₆) δ 9.35 (d, J= 9.8 Hz, 1H), 9.04 (s, 1H), 8.26 (s, 1H), 5.32 (s, 1H), 4.73 (s, 1H), 4.48 (s, 2H), 4.09 (s, 3H), 4.01 (s, 2H), 3.62 (s, 1H), 2.73 (s, 2H), 2.34 (s, 2H), 1.91 (d, J= 13.5 Hz, 1H), 1.29 (d, J= 6.5 Hz, 3H). LCMS m/z 403.13 [M+H]⁺.

Compound18

2-[4-[(2S,6S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-2'yl]pyrazol-l-yl]-N,N-dimethyl-acetamide (18)

To a solution of (2S,6S)-2-chloro-2'-methyl-6'-(lH-pyrazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] S31 (20 mg, 0.05865 mmol) in DMF (280 pL) was added CS2CO3 (57 mg, 0.1749 mmol). 2-bromo-N,N-dimethyl-acetamide (7.6 pL, 0.07050 mmol) was added at room température. The reaction was stirred for 1 hour._The reaction was quenched with saturated NaHCCL solution and extracted with EtOAc (4x). The combined organic layer was dried over Na2SÛ4, filtered, and concentrated. Purification by reversed-phase HPLC (Method: Cl8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O) 186 afforded 2-[4-[(2S,6S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4^,-piperidine]2'-yl]pyrazol-l-yl]-N,N-dimethyl-acetamide 18 (6.2 mg, 23%). 'H NMR (300 MHz, Chloroform-iO δ 7.50 (s, 2H), 6.57 (s, 1H), 4.92 (s, 2H), 4.17 (dd, J= 11.6, 2.5 Hz, 1H), 3.93 (t, J= 5.5 Hz, 2H), 3.25 (d, J= 9.1 Hz, 1H), 3.06 (s, 3H), 2.97 (s, 3H), 2.60 (td, J= 5.4, 1.8 Hz, 2H), 2.25 (d, .7=13.6 Hz, 1H), 2.01 (s, 1H), 1.70 (d, .7= 12.5 Hz, 1H), 1.47 - 1.32 (m, 1H), 1.11 (d, J= 6.4 Hz, 3H). LCMS m/z 409.19 [M+H]⁺.

Compound 19 (2S)-2-chloro-2 '-methyl-6'-(l-methylpyrazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 piperidine] (19)

L-Proline TEA

MgSO₄

TfOH

A solution of (45)-4-aminopentan-2-one hydrochloride S25 (25 mg, 0.1817 mmol) and TEA (30 pL, 0.2152 mmol) in MeCN (1.000 mL) was added to l-methylpyrazole-4carbaldehyde (22.01 mg, 0.20 mmol), MgSCri (25 mg, 0.2077 mmol), and L-proline (5 mg, 0.043 mmol). The resulting mixture was stirred at room température ovemight. The reaction mixture was evaporated via Genevac at 40 °C until dry to afford crude C58. To this, a solution of 2-(5chloro-3-thienyl)ethanol S2 (25 pL, 0.2080 mmol) in dioxane (750.0 pL) was added, followed by a solution of TfOH (80 pL, 0.90 mmol) in dioxane (750.0 pL). The mixture was stirred at room température for 30 minutes. Additional triflic acid (50 pL, 0.5650 mmol) was added and stirring was continued for 10 minutes. The reaction was placed under a nitrogen stream until the volume was reduced by half. The remaining solution was quenched with NaOH (1.5 mL of 2 M, 3.000 mmol) and diluted with DCM (1.500 mL). The resulting biphasic mixture was stirred for several minutes and then passed through a phase separator. The organic layer was blown down with nitrogen. Purification by reversed-phase HPLC (Method: Cl8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H₂O with 0.1% trifluoroacetic acid) afforded (2S)-2chloro-2'-methyl-6'-(l-methylpyrazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] 19 as a trifhioroacetate sait (6.6 mg, 11%). Compound 19 was determined to be 88% e.r. by chiral SFC analysis (Method: AD-H column (4.6 x 100 mm). Gradient: 10% MeOH with 5 mM ammonia with 90% CO₂). ‘H NMR (400 MHz, DMSO-î/₆) δ 8.91 (d, 10.8 Hz, 1H), 8.49 (d, J = 11.3 Hz, 1H), 7.86 (s, 1H), 7.59 (s, 1H), 6.94 (s, 1H), 4.49 (t, J= 11.2 Hz, 1H), 3.93 (t, J= 5.5 187

Hz, 2H), 3.84 (s, 3H), 2.93 (td, J = 13.9, 6.9 Hz, 1H), 2.60 (t, J= 5.5 Hz, 2H), 2.35 (d, J= 17.2

Hz, 1H), 2.21 (q, J= 13.9 Hz, 2H), 1.84 - 1.73 (m, 1H), 1.24 (d, J = 6.6 Hz, 3H). LCMS m/z 338.17 [M+H]⁺

Compound 20

2-chloro-2 '-methyl-6'-(3-pyridyl)spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine] (20)

A solution of 4-aminopentan-2-one hydrochloride S24 (25 mg, 0.1817 mmol) in EtOH (1 mL) was added to pyridine-3-carbaldehyde (19.5 mg, 17.06 pL, 0.1817 mmol), MgSCU (25 mg, 0.2077 mmol), and L-proline (5 mg, 0.04343 mmol). TEA (30 pL, 0.2152 mmol) was added and the reaction was stirred at room température over 3 days. The reaction mixture was evaporated under a stream of nitrogen to afford crude C59. To this, a solution of 2-(5-chloro-3thienyl)ethanol S2 (25 pL, 0.2075 mmol) in dioxane (750 pL) was added, followed by a freshly prepared solution of TfOH (100 pL, 1.130 mmol) in dioxane (750 pL). The mixture was stirred at room température for 30 minutes. The reaction was placed under a nitrogen stream until the volume was reduced by half. The remaining solution was quenched with NaOH (1.5 mL of 2 M, 3.000 mmol) and diluted with DCM (1.500 mL). The resulting biphasic mixture was stirred for several minutes and then passed through a phase separator. The organic layer was blown down with nitrogen. Purification by reversed-phase HP LC (Method: Cl 8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 0.1% trifluoroacetic acid) afforded 2-chloro2'-methyl-6'-(3-pyridyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]20 as a trifluoroacetate sait (34.4 mg, 56%). Compound 20 was determined to be 94% cis enantiomers and 6% trans enantiomers by chiral SFC analysis (Method: AD-H column (4.6 x 100 mm). Gradient: 10% MeOH with 5 mM ammonia with 90% CO2). *H NMR (300 MHz, Methanol-i/4) 8 8.87 (d, J =2.3 Hz, 1H), 8.74 (dd, J =3.2, 1.5 Hz, 1H), 8.36 - 8.27 (m, 1H), 7.77 (dd, 7=8.1, 5.2 Hz, 1H), 6.75 (s, 1H), 4.93 - 4.88 (m, 1H), 4.03 (t, J= 5.5 Hz, 2H), 3.90 (dqd, J= 13.4, 6.7, 3.1 Hz, 1H), 2.67 (t, J= 5.6 Hz, 2H), 2.51 (dt, J= 14.5, 2.9 Hz, 1H), 2.46 - 2.30 (m, 2H), 1.93 (dd, J= 14.8, 12.2 Hz, 1H), 1.41 (d, J= 6.6 Hz, 3H). LCMS m/z 335.14 [M+H]⁺.

188

Compound 21 (2S)-2-chloro-2 '-methyl-6'-(2-methyl-4-pyridyl)spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 piperidine] (21)

A solution of (45)-4-aminopentan-2-one hydrochloride S25 (34.40 mg, 0.2500 mmol) in EtOH (1 mL) was added to 2-methylpyridine-4-carbaldehyde (30.28 mg, 0.2500 mmol), MgSCU (45 mg, 0.3739 mmol), and L-proline (7 mg, 0.06080 mmol). TEA (40 pL, 0.2870 mmol) was added and the reaction was stirred at room température ovemight. The reaction mixture was evaporated via Genevac between 35-40 °C to afford crude C60. To C60, a solution of 2-(5chloro-3-thienyl)ethanol S2 (35 pL, 0.2905 mmol) in dioxane (1 mL) was added, followed by a freshly prepared solution of TfOH (130 pL, 1.469 mmol) in dioxane (1 mL). The mixture was stirred at room température for 30 minutes. The reaction mixtures were evaporated via Genevac at 40 °C. The residue was quenched with NaOH (1.7 mL of 2 M, 3.400 mmol) and diluted with DCM (1.7 mL). The resulting biphasic mixture was stirred for several minutes and then passed through a phase separator. The organic layer was blown down with nitrogen. Purification by reversed-phase HPLC (Method: Cl8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 0.1% trifluoroacetic acid) afforded (2S)-2-chloro-2'-methyl-6'-(2methyl-4-pyridyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] 21 as a trifluoroacetate sait (19.3 mg, 21%). Compound 21 was determined to be 77% e.r. by chiral SFC analysis (Method: AD-H column (4.6 x 100 mm). Gradient: 10% MeOH with 5 mM ammonia with 90% CO₂). Ή NMR (400 MHz, DMSOé₆) δ 9.28 (s, 1H), 8.87 (s, 1H), 8.55 (d, J= 5.3 Hz, 1H), 7.51 (s, 1H), 7.42 (d, 7=5.3 Hz, 1H), 6.94 (s, 1H), 4.57 (t, 7= 11.5, 9.8 Hz, 1H), 3.97 (t, 7= 5.4 Hz, 2H), 3.6 (1H hiddenunder water peak), 3.17 - 2.84 (m, 1H), 2.61 (q, 7= 5.3 Hz, 2H), 2.52 (s, 3H), 2.43-2.13 (m, 3H), 1.90 (t, 7= 13.3 Hz, 1H), 1.29 (d, 7= 6.4 Hz, 3H). LCMS m/z 349.14 [M+H]+

Compounds 22-172

Compounds 22-172 (see Table 3) were prepared as trifluoroacetate salts in a two-step, one pot procedure following the methods described for compounds 19, 20, or 21. Intermediate S24 or S25, appropriate aldéhyde, and thiophene éthanol S2 were used. Aldéhydes were

189 prepared by methods described above or obtained from commercial sources. Partial stereochemical érosion of the enantiomerically pure starting material (45)-4-aminopentan-2-one (Hydrochloride sait) S25 was observed under step 1 reaction conditions, leading to unseparated mixtures of enantiomers of the 2,6-trans piperidines. This results from the mixture of cis5 piperidinone intermediates (as previously described in Method A préparation of S26) and subséquent 2,6 trans piperidine final products, in which the 2 and 6 substituents are cis, and the 2 and 4 substituents are trans. Any modifications to methods are noted in Table 3 and accompanying footnotes.

Table 3. Method of préparation, structure and physicochemical data for compounds 22-172

Product	Starting Material and Aldéhyde	Method	’HNMR; LCMS m/z [M+H]'
Compound 22 \ Π H /	S25; 0 </ /	Compound 19^l	‘H NMR (400 MHz, DMSO-cZ₆) δ 7.55 (s, 1H), 7.07 (s, 1H), 6.89 (s, 1H), 4.12 (dd, J= 11.9, 2.7 Hz, 1H), 3.90 (t, J= 5.5 Hz, 2H), 3.61 (s, 3H), 3.30-3.15 (m, 1H), 2.57 (t,J= 5.4 Hz, 2H), 2.22 (d, J= 13.9 Hz, 1H), 2.10-2.01 (m, 1H), 1.79 (t,J= 12.9 Hz, 1H), 1.46 (t,J= 12.6 Hz, 1H), 1.10 (d,J= 6.4 Hz, 3H). LCMS m/z 338.17 [M+H]⁺.
Compound 23 ₀O>^{ci s} ⁰⁼\ L IJ h /	S25; 0 /	Compound 20	‘H NMR (300 MHz, DMSO-J₆) δ 9.02 (s, 1H), 8.42 (s, 1H), 7.60 (d, .7= 1.5 Hz, 1H), 7.487.41 (m, 1H), 7.35 (d, J = 8.1 Hz, 1H), 6.94 (s, 1H), 4.61 (s, 1H), 3.96 (t, .7=5.4 Hz, 2H), 3.64 (s, 1H), 3.36 (s, 3H), 2.61 (t, J =5.4 Hz, 2H), 2.34 - 2.23 (m, 3H), 1.87 (t, J =13.3 Hz, 1H), 1.28 (d, .7= 6.5 Hz, 3H). LCMS m/z 405.13 [M+H]⁺.
Compound 24 (OlV^ci Γ J N ^H	S25; iQi	Compound 21	Ή NMR (400 MHz, DMSO-ώί) δ 9.12 (s, 1H), 8.68 (s, 1H), 8.10 (s, 1H), 7.94-7.89 (m, 2H), 7.68 (t, .7= 7.8 Hz, 1H), 6.94 (s, 1H), 4.70 - 4.60 (m, lH),3.97(t, .7= 5.5 Hz, 2H), 3.65 (s, 1H), 2.61 (q, .7=5.0 Hz, 2H), 2.41 - 2.22 (m, 3H), 1.89 (t, J= 13.3 Hz, 1H), 1.28 (d, J= 6.5 Hz, 3H). LCMS m/z 359.12 [M+H]⁺.

190

Product	Starting Material and Aldéhyde	Method	‘HNMR; LCMS m/z [M+Hf
Compound 25 oOr^^c\|“Xj'S ίι^Υ N Π Ί ^H 1 N-O	S25; 0 H N-0	Compound 21	Ή NMR (400 MHz, DMSOd₆) δ 9.25 (s, 1H), 8.75 (s, 1H), 8.00 - 7.94 (m, 2H), 7.60 (d, J = 8.3 Hz, 1H), 6.94 (s, 1H), 4.76 (t, 7= 10.8 Hz, 1H), 3.99 (t, 7=5.5 Hz, 2H), 3.68 (s, 1H), 2.62 (d, 7=6.0 Hz, 2H), 2.57 (s, 3H), 2.42 - 2.26 (m, 3H), 1.93 (t, 7= 13.3 Hz, 1H), 1.30 (d, 7= 6.5 Hz, 3H). LCMS m/z 389.15 [M+H]⁺.
Compound 26 □CO-c. fXf ^sfil h ~~~N N=^	S25; O I] ï 1 N N=/	Compound 21	*H NMR (300 MHz, Methanol7₄)δ8.72 (d, 7=2.2 Hz, 1H), 8.41 (d, 7=2.1 Hz, 1H), 8.16 (s, 1H), 6.77 (s, 1H), 4.96 (dd, 7= 12.5, 2.8 Hz, 1H), 4.13 (s, 3H), 4.05 (t, 7= 5.4 Hz, 2H), 3.92 (s, 1H), 2.68 (t,7=5.7 Hz, 2H), 2.55 (dd, 7= 14.3,2.3 Hz, 1H), 2.43 (dtd, 7= 12.7, 7.3, 6.3, 2.5 Hz, 2H), 1.89 (t, 7 = 13.6 Hz, 1H), 1.41 (d, J =6.6 Hz, 3H). LCMS m/z 389.15 [M+H]⁺.
Compound 27 ₀Ο>°ι ^U><L S ⁰ il H [Il H	S25; 0 0 ÏÏ H τί^τ H	Compound 21	Ή NMR (400 MHz, DMSOd₆) δ 9.09 (d, 7= 10.0 Hz, 1H), 8.66 - 8.50 (m, 2H), 8.05 (s, 1H), 7.87 (d, 7= 7.8 Hz, 1H), 7.70 (d, 7=7.7 Hz, 1H), 7.55 (t, 7=7.7 Hz, 1H), 6.95 (s, 1H), 4.60 (t, 7= 11.1 Hz, 1H), 3.97 (t, 7= 5.4 Hz, 2H), 3.66 (s, 1H), 2.81 (d, 7= 4.4 Hz, 3H), 2.65 -2.58 (m, 2H), 2.31 (dd, 7=28.1, 14.2 Hz, 3H), 1.90 (t, 7= 13.3 Hz, 1H), 1.28 (d, 7= 6.5 Hz, 3H). LCMS m/z 391.13 [M+H]⁺.
Compound 28 ₀OlV^ci~>ς3 \ Il ^O=K L U h	S25; \ jj X J	Compound 21	^rH NMR (400 MHz, DMSOdé) δ 9.16 (s, 1H), 8.68 (s, 1H), 7.56 - 7.27 (m, 3H), 6.94 (s, 1H), 4.59 (s, 1H), 3.96 (t, 7 = 5.4 Hz, 2H), 3.63 (s, 1H), 3.36 (s, 3H), 2.62 (d, 7=6.1 Hz, 2H), 2.28 (d, 7= 15.6 Hz, 3H), 1.90 (t, 7= 13.3 Hz, 1H), 1.28 (d, 7= 6.4 Hz, 3H). LCMS m/z 405.13 [M+H]⁺.

191

Product	Starting Material and Aldéhyde	Method	Ή NMR; LCMS m/z [M+Hf
Compound 29 oOlV^ciJJ s \ M ^H O-N	S25; 0 H O-N	Compound 19	‘H NMR (400 MHz, DMSOd₆) δ 9.55 (s, 1H), 9.07 (s, 1H), 6.95 (s, 1H), 6.49 (s, 1H), 4.77 - 4.66 (m, 1H), 4.01 -3.93 (m, 3H), 2.62 (t, J= 5.5 Hz, 2H), 2.51 (d, 7=14.3 Hz, peak obscured by DMSO solvent, 1H), 2.45 (s, 3H), 2.27 (d, J= 14.3 Hz, 1H), 2.06 (t, J= 13.4 Hz, 1H), 1.83 (t, J= 13.3 Hz, 1H), 1.29 (d, J =6.5 Hz, 3H). LCMS m/z 339.21 [M+H]⁺.
Compound 30 oOl^^ciJJ S M ^H	S25; 0 H iXX	Compound 19²	LCMS m/z 335.19 [M+H]⁺.
Compound 31 oOO^^clJJs ^ν>Ό h	S24; 0 H N J HN^	Compound 19¹’³’⁴	Ή NMR (300 MHz, Chloroformé) δ 7.57 (s, 2H), 6.57 (s, 1H), 4.22 (dd,7=11.7, 2.5 Hz, 1H), 3.94 (t, J= 5.5 Hz, 2H), 3.28 (dtd, 7=11.6, 6.1, 5.3,3.6 Hz, 1H), 2.61 (t, J= 5.4 Hz, 2H), 2.24 (dt, J= 13.7, 2.6 Hz, 1H), 2.07 (dt, 7= 13.8, 2.5 Hz, 1H), 1.74 (dd,7= 13.7, 11.7 Hz, 1H), 1.44 (dd,7 = 13.8, 11.4 Hz, 1H), 1.14 (d, 7= 6.4 Hz, 3H). LCMS m/z 324.02 [M+H]⁺.
Compound 32 oOO^^ci ~X^S \ 11 N^\A_nA. O=< X) H /	S24; 0 \ J °=< X Y !	Compound ₁₉1.3,4	¹H NMR (300 MHz, Chloroformé) δ 7.13 (d, 7= 10.8 Hz, 2H), 6.88 (d,7=7.9 Hz, 1H), 6.58 (s, 1H), 4.25 (d, 7=11.5 Hz, 1H), 3.97 (t, 7= 5.4 Hz, 2H), 3.49-3.33 (m, 7H), 2.62 (t, 7= 5.4 Hz, 2H), 2.15 (dd, 7= 23.1, 13.7 Hz, 2H), 1.98 - 1.74 (m, 2H), 1.18 (d, 7= 6.2 Hz, 3H). LCMS m/z 418.12 [M+H]⁺.

192

Product	Starting Material and Aldéhyde	Method	'H NMR; LCMS m/z [M+Ilf
Compound 33 (OüVci éx ^sX X iij ^H F	S25; N^H° Xx F	Compound 20	’H NMR (300 MHz, Methanolί/4) δ 7.78 (d, J= 1.5 Hz, 1H), 7.74 - 7.66 (m, 2H), 6.76 (s, 1H), 4.81 (dd, J= 3.0 Hz, under water peak, 1H), 4.03 (t, J= 5.5 Hz, 2H), 3.87 (dtq, J= 12.8, 6.5, 2.9 Hz, 1H), 2.67 (t, .7= 5.5 Hz, 2H), 2.51 (dt, J= 14.6, 2.8 Hz, 1H), 2.45 - 2.37 (m, 1H), 2.28 -2.17 (m, 1H), 1.88 (dd, J= 14.8, 12.3 Hz, 1H), 1.41 (d, J =6.6 Hz, 3H). LCMS m/z 377.14 [M+H]⁺.
Compound 34 oOl^^ciyxf'' ^s Il J h	S25; 0 XJ	Compound 20	Ή NMR (400 MHz, DMSOé) δ 9.28 (d, J= 10.5 Hz, 1H), 8.82 (d, .7=11.3 Hz, 1H), 8.37 (d, .7= 2.8 Hz, 1H), 7.60-7.45 (m, 2H), 6.94 (s, 1H), 4.63 (t, J = 11.2 Hz, 1H), 3.98 (hept, J = 5.9,5.4 Hz, 2H), 3.86 (s, 3H), 3.58 (s, 1H), 2.61 (t, .7= 5.5 Hz, 2H), 2.46-2.18 (m, 2H), 1.98 (ddd, J= 65.1, 14.4, 12.3 Hz, 2H), 1.30 (d, J =6.5 Hz, 3H). LCMS m/z 365.17 [M+H]⁺.
Compound 35 OlV^ciéx ^S II· Ί H H N=^	S25; 0 H rv H N	Compound 20	Ή NMR (400 MHz, DMSOé) δ 13.25 (s, 1H), 9.07 (d, J= 10.7 Hz, 1H), 8.55 (d, J= 11.5 Hz, 1H), 8.16 (s, 1H), 7.99 (s, 1H), 7.62 (d, .7= 8.6 Hz, 1H), 7.54 (dd, J =8.7, 1.6 Hz, 1H), 6.94 (s, 1H), 4.65 (q, J=8.8 Hz, 1H), 3.97 (t, .7= 5.5 Hz, 2H), 3.67 (s, 1H), 2.61 (q, J= 4.8 Hz, 2H), 2.32 (dd, J= 23.3, 11.3 Hz, 3H), 1.97- 1.85 (m, 1H), 1.28 (d, J= 6.5 Hz, 3H). LCMS m/z 374.16 [M+H]⁺.

193

Product	Starting Material and Aldéhyde	Method	‘HNMR; LCMS m/z [M+Ilf
Compound 36 IM ^H ^0 0 \	S25; 0 IJ ifY C)^ ^0 Ό \	Compound 21	Ή NMR (400 MHz, DMSOé) 5 9.02 (d, J= 10.7 Hz, 1H), 8.49 (s, 1H), 7.49 (d, J= 8.5 Hz, 2H), 7.02 (d, .7=8.3 Hz, 2H), 6.93 (s, 1H), 4.53-4.43 (m, 1H), 4.11 (dd, J= 5.7, 3.6 Hz, 2H), 3.95 (t,J= 5.4 Hz, 2H), 3.76 - 3.69 (m, 2H), 3.67 3.60 (m, 1H), 3.58 (dd, J= 5.7, 3.7 Hz, 2H), 3.48 -3.42 (m, 2H), 3.24 (s, 3H), 2.64-2.57 (m, 2H), 2.30 - 2.22 (m, 3H), 1.87 (t,J = 13.3 Hz, 1H), 1.27 (d, J= 6.5 Hz, 3H). LCMS m/z 365.17 [M+H]⁺.
Compound 37 _oQ[V^ciSM Ο N'MM'M ÏÏ jl \ H H	S25; 0 J 0 N'^ ÏÏ ï \ H	Compound 21	‘H NMR (300 MHz, DMSOd₆) δ 10.65 (s, 1H), 8.94 (s, 1H), 8.52 (s, 1H), 8.48 (d, J = 2.4 Hz, 1H), 8.13 (d, .7= 8.7 Hz, 1H), 7.96 (dd, J= 8.7, 2.5 Hz, 1H), 6.94 (s, 1H), 4.58 (s, 1H), 3.96 (t, J =5.4 Hz, 2H), 3.64 (s, 1H), 2.61 (s, 2H), 2.34 - 2.21 (m, 3H), 2.10 (s, 3H), 1.92- 1.80 (m, 1H), 1.27 (d, J = 6.5 Hz, 3H). LCMS m/z 392.13 [M+H]⁺.
Compound 38 <OlV^ci X' S ~K 2 H M	S25; 0 Π N^ïl N'N /	Compound 21	Ή NMR (400 MHz, DMSOd₆) δ 9.37 (s, 1H), 8.94 (s, 1H), 7.92 (s, 1H), 6.95 (s, 1H), 4.72 (t, .7=11.4 Hz, 1H), 4.19 (s, 3H), 3.95 (t, J =5.4 Hz, 2H), 3.67-3.55 (m, 1H), 2.61 (t, J = 5.4 Hz, 2H), 2.47 - 2.45 (m, 1H), 2.31 -2.15(m, 2H), 1.83 (dd, J= 14.5, 12.3 Hz, 1H), 1.27 (d, J =6.4 Hz, 3H). LCMS m/z 339.16 [M+H]⁺.
Compound 39 Î^^V^—ci ^ox?^s „9 Μ M 0	S25; 0 1] jfj o^°	Compound 21	Ή NMR (400 MHz, DMSOJô) δ 10.86 (s, 1H), 9.03 (d, J= 10.1 Hz, 1H), 8.63 - 8.31 (m, 1H), 7.21 (d,J=1.8Hz, 1H), 7.16-7.10 (m, 1H), 6.93 (t, J = 4.1 Hz, 2H), 4.60 (s, 2H), 4.45 (q, .7=10.2, 9.6 Hz, 1 H), 3.94 (t, J =5.4 Hz, 2H), 3.60 (s, 1H), 2.59 (t,J= 5.3 Hz, 2H), 2.29-2.19 (m, 3H), 1.92-1.80

194

Product	Starting Material and Aldéhyde	Method	Ή NMR; LCMS m/z [M+llf
			(m, 1H), 1.26 (d, 7= 6.5 Hz, 3H). LCMS m/z 405.09 [M+H]⁺.
Compound 40 oOl^^ci ^Nfj) H 4 7 N-N	S25; 0 J \\ v	Compound 21	Ή NMR (400 MHz, DMSOd₆) δ 9.19- 9.07 (m, 1H), 8.81 (s, 1H), 8.78 - 8.70 (m, 1H), 8.60 - 8.55 (m, 1H), 8.25 - 8.20 (m, 1H), 8.02 (s, 1H), 7.59 (dd, 7=8.4, 4.7 Hz, 1H), 6.96 (s, 1H), 4.61 (t, 7= 11.2 Hz, 1H), 3.96 (t, 7= 5.5 Hz, 2H), 3.62 (s, 2H), 2.62 (t, 7= 5.2 Hz, 2H), 2.48 -2.43 (m, 1H), 2.31 (dd, 7=27.8, 13.8 Hz, 2H), 1.84 (t, 7= 13.3 Hz, 1H), 1.28 (d, J= 6.4 Hz, 3H). LCMS m/z 401.11 [M+H]⁺.
Compound 41 oOlV^ci ^Sajc// O	S25; o 0	Compound 21	Ή NMR (400 MHz, DMSOd₆} δ 9.17 (s, 1H), 8.69 (s, 1H), 8.54 (d, 7= 4.8 Hz, 1H), 7.91 (d, J= 8.0 Hz, 2H), 7.65 (d, 7= 8.1 Hz, 2H), 6.94 (s, 1H), 4.61 (t, 7= 11.2 Hz, 1H), 3.97 (t,7= 5.5 Hz, 2H), 3.65 (s, 1H), 2.79 (d, 7=4.4 Hz, 3H), 2.61 (s, 2H), 2.39-2.19 (m, 3H), 1.91 (t, 7= 13.3 Hz, 1H), 1.29 (d,7 = 6.4 Hz, 3H). LCMS m/z 391.13 [M+H]⁺.
Compound 42 oOlV⁰¹JxJ'S Γ JJ H -NT N=\	J Z CO II // \\ / —\ / 5*¹ \=o	Compound 21¹	‘H NMR (400 MHz, DMSOd₆) δ 8.52 (d, 7= 1.9 Hz, 1H), 8.21 (s, 1H), 6.86 (s, 1H), 4.21 (d, 7= 11.4 Hz, 1H), 3.95 (s, 5H), 3.18 (s, 1H), 2.58 (t, 7= 5.4 Hz, 2H), 2.49 (s, 3H),2.11 (d, 7= 13.2 Hz, 1H), 2.03 (d,7 = 13.6 Hz, 1H), 1.67 (s, 1H), 1.35 (t, 7= 13.1 Hz, 1H), 1.06 (d, 7= 6.3 Hz, 3H). LCMS m/z 403.14 [M+H]⁺.

195

21175'

Product	Starting Material and Aldéhyde	Method	¹II.NMR; LCMS m/z [M+Hf
Compound 43 oCcyc AA υΆΑ nJ ^h	S25; 0 .nA II Ύ ^νΆ	Compound 21	‘H NMR (400 MHz, DMSOtZd) δ 9.53 (s, lH),9.16(d, J= 1.4 Hz, 1H), 9.04 (d, J= 10.5 Hz, 1H), 7.65 (s, 1H), 6.95 (s, 1H), 4.74 - 4.61 (m, 1H), 4.01 (m, 2H), 3.66 - 3.52 (m, 1H), 2.63 (t, J =5.5 Hz, 2H), 2.57 (s, 1H), 2.50 (s, 3H), 2.25 (d, J = 14.4 Hz, 1H), 1.94 (dt, J= 30.8, 13.5 Hz, 2H), 1.33 (d, J= 6.4 Hz, 3H). LCMS m/z 350.14 [M+H]⁺.
Compound 44 OlV^ci f II AAn^ Il Ί h	S25; F 0 Ί H Jlj	Compound 21	LCMS m/z 382.1 [M+H]⁺.
Compound 45 oOlV^ciA<As 2 ^H N	S25; O l\| Ù J	Compound 19¹¹	Ή NMR (400 MHz, DMSOd₆) δ 9.24 (s, 1H), 9.16 (s, 1H), 9.01 (s, 2H), 8.88 (s, 1H), 6.95 (s, 1H), 4.71 (s, lH),3.97(t, J = 5.5 Hz, 2H), 3.66 (s, 1H), 2.61 (t, 7= 4.9 Hz, 2H), 2.41 (d, 7= 11.7 Hz, 2H), 2.29 (d,7 = 14.4 Hz, 1H), 1.93 - 1.84 (m, 1H), 1.29 (d, 7=6.7 Hz, 3H). LCMS m/z 336.18 [M+H]⁺.
Compound 46 <OTA^cis A} h γΉ o^sC^ /'O	S25; 0 H N'A/ A° S19	Compound 19	Ή NMR (400 MHz, DMSOd₆) δ 8.95 (d, 7= 10.6 Hz, 1H), 8.51 (d, 7 = 11.3 Hz, 1H), 8.00 (s, 1H), 7.69 (s, 1H), 6.95 (s, 1H), 4.54 (q, 7=9.0, 7.9 Hz, 3H), 3.93 (t, 7=5.5 Hz, 2H), 3.68 (t, 7= 6.7 Hz, 2H), 3.57 (s, 1H), 2.86 (s, 3H), 2.64 2.56 (m, 2H), 2.38-2.31 (m, 1H), 2.27-2.14 (m, 2H), 1.83 1.70 (m, 1H), 1.25 (d, J =6.5 Hz, 3H). LCMS m/z 430.1 [M+H]⁺.

196

Product	Starting Material and Aldéhyde	Method	¹HNMR; LCMS m/z [M+H]
Compound 47 oOC^^ci HN jO « 0	o=\__ iri / CM '---( «> )=o Z CM T	Compound 20	‘H NMR (300 MHz, Methanol</4)67.99 (d, J =8.4 Hz, 2H), 7.63 (d, J =8.2 Hz, 2H), 6.76 (s, 1H), 4.77 (dd, J= 12.7, 2.8 Hz, 1H), 4.03 (t, J=5.4 Hz, 2H), 3.89 (dt, J= 11.3, 7.8 Hz, 1H), 2.67 (t, .7= 5.5 Hz, 2H), 2.52 - 2.37 (m, 2H),2.31 -2.18 (m, 1H), 1.89 (t,J= 13.5 Hz, 1H), 1.41 (d, .7=6.6 Hz, 3H). LCMS m/z 377.18 [M+H]⁺.
Compound 48 oOl^^ciXX s ν^ζ%Χ*'·_νΑ H,V ^H N=\	S25; 0 JJ nAZ jiJ H N N=^X	Compound 20	Ή NMR (400 MHz, DMSOdè) δ 13.37 (s, 1H), 9.07 (d, J= 10.7 Hz, 1H), 8.71 (s, 1H), 8.66 (d, .7= 2.1 Hz, 1H), 8.47 (d,J= 2.1 Hz, 1H), 6.95 (s, 1H), 4.77 (t, .7=11.2 Hz, 1H), 3.98 (t,J= 5.4 Hz, 2H), 3.68 (s, 1H), 2.64 - 2.56 (m, 2H), 2.52 (s, 3H), 2.48 - 2.25 (m, 3H), 1.92 (t, J = 13.3 Hz, 1H), 1.28 (d,J=6.5 Hz, 3H). LCMS m/z 389.19 [M+H]⁺.
Compound 49 (OC^^ci SAs Ji \ H	S25; 0 JJ nAZ ï J	Compound 20	Ή NMR (300 MHz, Methanold]) δ 8.30 (d, J= 2.6 Hz, 1H), 7.85 (dd, J= 8.7, 2.6 Hz, 1H), 6.90 (d, .7=8.7 Hz, 1H), 6.75 (s, 1H), 4.71 (dd,J= 12.5,3.0 Hz, 1H), 4.02 (t, J= 5.5 Hz, 2H), 3.93 (s, 3H), 3.85 (dddd, J = 15.6,9.1,6.9,3.3 Hz, 1H), 2.66 (t, J =5.5 Hz, 2H), 2.482.22 (m, 3H), 1.87 (dd, J= 14.9, 12.3 Hz, 1H), 1.38 (d,J= 6.6 Hz, 3H). LCMS m/z 365.17 [M+H]⁺.
Compound 50 O0^^{cl s} ο j II J ^H H	S25; o H A jo H	Compound 20	*H NMR (400 MHz, DMSOdè) δ 10.09 (s, 1H), 9.04 (s, 1H), 8.55 (s, 1H), 7.64 (d,J= 8.4 Hz, 2H), 7.48 (d, .7=8.3 Hz, 2H), 6.94 (s, 1H), 4.53 4.42 (m, 1H), 3.96 (t,J= 5.1 Hz, 2H), 3.67 - 3.58 (m, 1H), 2.60 (t, 2H), 2.26 (d, J =7.8 Hz, 3H), 2.05 (s, 3H), 1.88 (t, J = 13.4 Hz, 1H), 1.27 (d, J =6.5 Hz, 3H). LCMS m/z 391.13 [M+H]⁺.

197

Product	Starting Material and Aldéhyde	Method	'il NMR; LCMS m/z [M+H]
Compound 51 oOl^“^ci Il Ί h HCT^	S25; O IJ HCT^	Compound 21	^!H NMR (400 MHz, DMSOJ_d) δ 10.25 (s, 1H), 9.00 (s, 1H), 8.51 (s, 1H), 7.41 (dd, 7= 12.5, 2.1Hz, 1H), 7.23-7.14 (m, 1H), 7.04 - 6.96 (m, 1H), 6.93 (s, 1H), 4.51-4.40 (m, 1H), 3.94 (t, 7= 5.3 Hz, 2H), 3.65 - 3.56 (m, 1H), 2.60 (s, 2H), 2.24 (d, 7= 10.4 Hz, 3H), 1.86 (t, 7= 13.4 Hz, 1H), 1.26 (d, 7= 6.5 Hz, 3H). LCMS m/z 368.11 [M+H]⁺.
Compound 52 ₀OZV^C\|°><< ^s nAt N^Z*''¹’ A^N ^H	S25; 0 Π N AA ^A'	Compound 21	Ή NMR (300 MHz, DMSOdc) δ 8.92 (s, 1H), 8.80 (d,7= 2.9 Hz, 2H), 8.63 (s, 1H), 6.95 (s, lH),4.62(s, lH),3.96(s, 2H), 3.64 (s, 1H), 2.61 (s, 2H), 2.40 - 2.20 (m, 4H), 1.86 (t, 7= 13.6 Hz, 1H), 1.30- 1.22 (m, 3H), 1.05 (ddt, 7= 23.3, 5.6, 2.9 Hz, 4H). LCMS m/z 376.14 [M+H]⁺.
Compound 53 °Ql>^c' ^nXa iij ^H	S25; n AA	Compound 21	Ή NMR (300 MHz, DMSOd₆) δ 9.41 (d, 7= 10.3 Hz, 1H), 9.07- 8.83 (m, 1H), 8.26 - 8.10 (m, 2H), 7.96 (dd,7=7.7, 1.4 Hz, 1H), 6.95 (s, 1H), 4.83 (t, J = 11.1 Hz, 1H), 4.01 (h, 7= 6.3 Hz, 2H), 3.61 (s, 1H), 2.63 (t,7 = 5.4 Hz, 2H), 2.58 - 2.54 (m, under DMSO, 1H), 2.27 (d, J= 14.0 Hz, 1H), 1.98 (dt,7= 27.4, 13.3 Hz, 2H), 1.33 (d,7= 6.5 Hz, 3H). LCMS m/z 360.12 [M+H]⁺.
Compound 54 i^^irV-ci ^s Il J H (Λγ V^H 0	S25; 0 l\| jfy V 0	Compound 21	Ή NMR (400 MHz, DMSOdé) δ 10.93 (s, 1H), 8.97 (s, 1H), 8.51 (s, 1H), 7.17-7.13 (m, 1H), 7.06 - 7.02 (m, 2H), 6.94 (s, 1H), 4.61 (s, 2H), 4.45 (t, 7= 11.0 Hz, 1H), 3.95 (t,7 = 5.5 Hz, 2H), 3.63 (s, 1H), 2.60 (t, 2H), 2.31 -2.19 (m,3H), 1.86 (t, J= 13.4 Hz, 1H), 1.25 (d, 7= 6.5 Hz, 3H). LCMS m/z 365.17 [M+H]⁺.

198

Product	Starting Material and Aldéhyde	Method	ΉNMR; LCMS m/z [M+Hf
Compound 55 <OA^ciαΟ ^s JD J H	S24; O H νΆΑ JD J	Compound 19¹’³’⁴	Ή NMR (300 MHz, . Chloroformé) δ 8.49 (d, J= 2.3 Hz, 1H), 7.68 (dd, 7=8.0, 2.4 Hz, 1H), 7.13 (d, 7= 8.0 Hz, 1H), 6.59 (s, 1H), 4.17 (dd, 7= 11.6, 2.4 Hz, 1H), 3.98 (t, 7 = 5.5 Hz, 2H), 3.38 -3.24 (m, 1H), 2.63 (t, 7= 5.5 Hz, 2H), 2.55 (s, 3H), 2.18-2.05 (m, 2H), 1.71 (dd, 7= 13.6, 11.6 Hz, 1H), 1.46 (dd, 7=13.7, 11.3 Hz, 1H), 1.14 (d, 7= 6.3 Hz, 3H). LCMS m/z 349.0 [M+H]⁺.
Compound 56 ₀O>^ci nJ} H A HO	S24; 0 1] ^NJj TBSO S20	Compound içl.3.4,5	‘H NMR (300 MHz, Chloroformé) δ 7.49 (s, 1H), 7.45 (s, 1H), 6.58 (s, 1H), 4.22 -4.14(m, 3H), 3.99 - 3.90 (m, 4H), 3.35-3.18 (m, 1H), 2.61 (t, 7= 5.4 Hz, 2H), 2.22 (dt, 7 = 13.8,2.6 Hz, 1H), 2.06 (dt,7= 13.8,2.5 Hz, 1H), 1.73 (t,7= 12.7 Hz, 1H), 1.48 - 1.38 (m, 1H), 1.13 (d, 7= 6.4 Hz, 3H). LCMS m/z 368.03 [M+H]⁺.
Compound 57 (OlV^ci ~xA^s nAA ï J H H₂N N	S25; 0 H N^A A J H₂N N	Compound 20	Ή NMR (400 MHz, DMSOd₆) δ 8.86 (d, 7= 10.7 Hz, 1H), 8.50 (s, 1H), 8.38 (s, 2H), 6.99 - 6.85 (m, 3H), 4.41 (t, 7= 11.1 Hz, 1H), 3.93 (t, 7= 5.6 Hz, 2H), 3.66 - 3.55 (m, 1H), 2.60 (q, 7= 4.9 Hz, 2H), 2.41 - 2.16 (m, 3H), 1.88 - 1.76 (m, 1H), 1.25 (d, 7= 6.5 Hz, 3H). LCMS m/z 351.17 [M+H]⁺.
Compound 58 <OA^cizC⁵ JD J ^H	S25; 0 A	Compound 21	^XH NMR (400 MHz, DMSO7_d)ô9.61 (s, 1H), 9.24 (d,7= 11.2 Hz, 1H), 7.81 (d,7=8.8 Hz, 1H), 7.70 (d, 7= 8.6 Hz, 1H), 6.95 (s, 1H), 4.86 (t, 7= 11.1 Hz, 1H), 4.01 (tt,7= 11.9, 6.3 Hz, 2H), 3.62 (s, 1H), 2.67 (s, 3H), 2.62 (dd, 7= 12.0, 6.1 Hz, 2H), 2.57 (m, 1H), 2.29 (d, 7= 14.5 Hz, 1H), 2.12-2.02 (m, 1H), 1.99- 1.90 (m, 1H), 1.34 (d, 7= 6.5 Hz, 3H). LCMS m/z 350.14 [M+H]⁺.

199

Product	Starting Material and Aldéhyde	Method	’HNMR; LCMS m/z [M+Ilf
Compound 59 oOO^^ciζΥγ Il Ί H	S25; O JL J hct^	Compound 21	Ή NMR (400 MHz, DMSOd₆) δ 10.35 (s, 1H), 9.23 (s, 1H), 8.77 (d, 7=11.9 Hz, 1H), 8.21 (d, 7= 2.8 Hz, 1H), 7.41 (d, 7= 8.5 Hz, 1H), 7.29 - 7.22 (m, 1H), 6.94 (s, 1H), 4.55 (t, 7 = 11.5 Hz, 1H), 3.98 (11,7=6.2 Hz, 2H), 3.56 (s, 1H), 2.67 2.59 (m, 2H), 2.37 (d, 7= 14.4 Hz, 1H), 2.24 (d, 7 = 14.3 Hz, 1H), 2.13 -2.01 (m, 1H), 1.951.85 (m, 1H), 1.29 (d,7=6.5 Hz, 3H). LCMS m/z 351.13 [M+H]⁺.
Compound 60 (OO^^ciγχγ-δ N^^nA x 1 zj ^HAli H	S25; 0 ïï nAZ ï J H	Compound 21	*H NMR (300 MHz, Methanold4) δ 8.42 (s, 2H), 6.76 (s, 1H), 4.66 -4.55 (m, 1H), 4.00 (t,7= 5.5 Hz, 2H), 3.81 (d, 7= 10.5 Hz, 1H), 2.93 (s, 3H), 2.66 (t, J = 5.4 Hz, 2H), 2.49 - 2.20 (m, 3H), 1.89- 1.75 (m, 1H), 1.37 (d, 7= 6.6 Hz, 3H). LCMS m/z 365.13 [M+H]⁺.
Compound 61 <OlV^ciy<ys N^T'hT z^A J ^H -νΎ^ν N	S25; 0 1) z^A -.αΑν rY	Compound 21	*H NMR (400 MHz, DMSOd₆) δ 9.10 (s, 1H), 8.91 (s, 3H), 8.44 (s, 1H), 8.06 (s, 1H), 6.95 (s, 1H), 4.65 (d, 7=8.9 Hz, 1H), 4.05 -3.95 (m, 2H), 3.91 (s, 3H), 3.66 (s, 1H), 2.662.58 (m, 2H), 2.41 (d,7=8.0 Hz, 2H), 2.28 (d, 7= 14.2 Hz, 1H), 1.90 (t, 7= 13.3 Hz, 1H), 1.28 (d, 7= 6.5 Hz, 3H). LCMS m/z 416.1 [M+H]⁺.
Compound 62 oQlV⁰¹“Υγ s Il Ί H Α^γ II N	S25; 0 H jJj O^Y ¹ lll N	Compound 21^1,6	‘H NMR (400 MHz, DMSO7₆) δ 7.74 - 7.64 (m, 2H), 7.17 (d, 7= 8.8 Hz, 1H), 6.85 (s, 1H), 4.05 - 3.89 (m, 3H), 3.89 (s, 3H), 3.09 (s, 1H), 2.61 2.55 (m, 2H), 2.00 (dd,7= 23.1, 13.5 Hz, 2H), 1.47 (t, 7= 12.4 Hz, 1H), 1.27 (t, 7= 12.4 Hz, 1H), 1.03 (d, 7=6.3 Hz, 3H). LCMS m/z 389.15 [M+H]⁺.

200

Product	Starting Material and Aldéhyde	Method	‘HNMR; LCMS m/z [M+lIf
Compound 63 oOO^^ci“XT'S 0	S25; 0 0	Compound 21	Ή NMR (400 MHz, DMSOdè) δ 9.55 (s, 1H), 9.25 (d,J= 1.5 Hz, 1H), 9.17 (s, 1H), 8.94 (d, J= 1.5 Hz, 1H), 8.38 (s, 1H), 7.95 (s, 1H), 6.95 (s, 1H), 4.97 (d, J= 12.0 Hz, 1H), 4.08 - 3.95 (m, 2H), 3.64 (s, 1H), 2.66-2.61 (m, 2H), 2.74 - 2.54 (m, 1H), 2.28 (d, J= 14.4 Hz, 1H), 2.16-2.05 (m, 1H), 1.94 (t, J= 13.3 Hz, 1H), 1.32 (d, J = 6.5 Hz, 3H). LCMS m/z 379.17 [M+H]⁺.
Compound 64 <Oû^^ciyxO^s L J h N	S25; O H ΊΟτ N	Compound 19⁷	Ή NMR (400 MHz, DMSOdè) δ 9.10 (s, 1H), 8.69 (d,J= 10.8 Hz, 1H), 8.57 (d, J=2.1 Hz, 1H), 8.49 (s, 1H), 7.89 (s, 1H), 6.94 (s, 1H), 4.61 (t, 1H), 3.97 (t, J= 5.5 Hz, 2H), 2.962.89 (m, 1H), 2.52 (d, under DMSO, 1H), 2.64 - 2.57 (m, 2H), 2.35 (s, 3H), 2.29 (d, J= 18.5 Hz, 2H), 1.94- 1.84 (m, 1H), 1.28 (d,J=6.3 Hz, 3H). LCMS m/z 349.19 [M+H]⁺.
Compound 65 <Or^^ci nQ) Pi tsH c5	S25; 0 H	Compound 19	*H NMR (400 MHz, DMSOdè) δ 8.97 (s, 1H), 8.70 (s, 1H), 8.61 (d, .7=12.6 Hz, 1H), 7.94 (s, 1H), 7.82 (d, J =8.0 Hz, 2H), 7.53 (t, J =7.8 Hz, 2H), 7.35 (t, J=7.4 Hz, 1H), 6.96 (s, 1H), 4.60 (t, J= 11.1 Hz, 1H), 3.96 (t,J= 5.4 Hz, 2H), 3.61 (s, 1H), 2.61 (d, J =6.2 Hz, 2H), 2.46 (d, under DMSO, 1H), 2.35 -2.24 (m, 2H), 1.81 (t, J= 13.3 Hz, 1H), 1.28 (d, J = 6.5 Hz, 3H). LCMS m/z 400.16 [M+H]⁺.
Compound 66 oOlVc ,o^A_KIAv ^NU/ H	S24; 0 ^NJT	Compound 19⁸	*H NMR (300 MHz, Methanol<74)6 6.76 (s, 1H), 6.51 (s, 1H), 4.98 (dd, J= 12.7, 3.1 Hz, 1H), 4.00 (t,J= 5.4 Hz, 2H), 3.83 (ddd, J= 12.3, 6.5, 2.9 Hz, 1H), 2.66 (t, J= 5.4 Hz, 2H), 2.60 (dt, J= 14.5, 2.8 Hz, 1H), 2.38 (dt, J= 14.8, 2.8 Hz, 1H), 2.31 (s, 3H), 2.23 (dd, J= 14.4,

201

Product	Starting Material and Aldéhyde	Method	’HNMR; LCMS m/z [Μ+ΗΓ
			12.6 Hz, 1H), 1.83 (dd, J= 14.7, 12.3 Hz, 1H), 1.39 (d, 7= 6.6 Hz, 3H). LCMS m/z 339.16 [M+H]⁺.
Compound 67 <OlV^ci II· 1 H V / n-nh	S25; 0 N-NH	Compound 20	Ή NMR (400 MHz, DMSOd₆) δ 13.33 (s, 1H), 9.15 (d,7= 9.1 Hz, 1H), 8.60 (d, J= 11.5 Hz, 1H), 8.12 (s, 1H), 7.85 (d, 7= 8.4 Hz, 1H), 7.76 (s, 1H), 7.30 (dd,7= 8.4, 1.4 Hz, 1H), 6.94 (s, 1H), 4.69 (t, 7= 10.8 Hz, 1H), 3.98 (t, J =5.5 Hz, 2H), 3.69 (d, 7 = 11.0 Hz, 1H), 2.63 - 2.59 (m, 2H), 2.43 - 2.24 (m, 3H), 1.92 (dd,7= 14.5, 12.2 Hz, 1H), 1.29 (d, J =6.5 Hz, 3H). LCMS m/z 374.16 [M+H]⁺.
Compound 68 oOt^^CI T JJ H Hcr^	t O Vz !Z) // \\ N» \ / R¹ \=o	Compound 21	Ή NMR (400 MHz, DMSO<Z₆) δ 11.82 (s, 1H), 8.89 (s, 1H), 8.49 (s, 1H), 7.63 (d,7= 9.8 Hz, 2H), 6.93 (s, 1H), 6.39 (d, 7=9.3 Hz, 1H), 4.39 (q,7= 6.2 Hz, 1H), 3.93 (t, J= 5.4 Hz, 2H), 3.57 (q, 7= 9.1, 6.8 Hz, 1H), 2.59 (t, 2H), 2.22 (d, J= 9.9 Hz, 3H), 1.81 (t, .7= 13.3 Hz, 1H), 1.25 (d, J =6.7 Hz, 3H). LCMS m/z 351.13 [M+H]⁺.
Compound 69 oOC^^cl'X/' s [JJ H	S25; 0 £ï	Compound 21	‘H NMR (400 MHz, DMSO70 δ 9.29 (d, 7= 10.5 Hz, 1H), 8.82 (d, J= 11.1 Hz, 1H), 8.36 (d, 7= 2.8 Hz, 1H), 7.57-7.45 (m, 2H), 6.94 (s, 1H), 4.62 (t, 7 = 11.2 Hz, 1H), 4.14 (q, 7= 7.0 Hz, 2H), 3.98 (q, 7= 5.8 Hz, 2H), 3.58 (s, 1H), 2.61 (t,7= 5.4 Hz, 2H), 2.40 (d, 7= 14.5 Hz, 1H), 2.25 (d, 7= 14.4 Hz, 1H), 2.12-2.01 (m, 1H), 1.981.81 (m, 1H), 1.35 (t, 7= 6.9 Hz, 3H), 1.29 (d, 7=6.5 Hz, 3H). LCMS m/z 379.12 [M+H]⁺.

202

Product	Starting Material and Aldéhyde	Method	¹HNMR; LCMS m/z [M+HJ*
Compound 70 (Oo^^c\| Vo O	S25; 0 H Vo	Compound 21	‘H NMR (400 MHz, DMSOd₆) δ 9.45 (s, lH),9.10(s, 1H), 8.09 - 7.99 (m, 2H), 7.61 - 7.55 (m, 3H), 7.52 (s, 1H), 6.97 (s, 1H), 4.86 (s, 1H), 3.97 (t, J = 5.5 Hz, 2H), 3.68 (s, 1H), 2.61 (d, 7= 14.1 Hz, 3H), 2.34 (t, J = 13.6 Hz, 1H), 2.27 (d, J= 14.5 Hz, 1H), 1.87 (t, J = 13.4 Hz, 1H), 1.29 (d, 7= 6.5 Hz, 3H). LCMS m/z 401.11 [M+H]⁺.
Compound 71 (OlV~^ci ^H NH	S25; o JJ \Z^^NH	Compound 21	’H NMR (400 MHz, DMSO7,5)0 9.19 (s, 1H), 8.71 (s, 1H), 8.52 (d, J= 4.3 Hz, 1H), 7.89 (d, J= 8.2 Hz, 2H), 7.63 (d, 7= 8.2 Hz, 2H), 6.93 (s, 1H), 4.60 (t, 7= 11.0 Hz, 1H), 3.97 (t,7= 5.5 Hz, 2H), 3.65 (s, 1H), 2.85 (dq, 7= 7.5, 3.7 Hz, 1H), 2.61 (s, 2H), 2.28 (h, 7= 13.5 Hz, 3H), 1.91 (t, 7 = 13.3 Hz, 1H), 1.29 (d, 7= 6.5 Hz, 3H), 0.70 (h, 7= 4.6 Hz, 2H), 0.58 (q, 7 = 3.8, 3.2 Hz, 2H). LCMS m/z 417.14 [M+H]⁺.
Compound 72 <OlV^ci“Xps Ν'J H HN^	S24; 0 Hbr	Compound 191,3,4,9	‘H NMR (300 MHz, Chloroform-7) δ 7.62 (s, 1H), 6.58 (s, 1H), 4.60 (dd,7= 12.1, 2.8 Hz, 1H), 3.94 (t, 7= 5.5 Hz, 2H), 3.58 - 3.50 (m, 1H), 2.61 (d, 7=3.1 Hz, 2H), 2.44-2.32 (m, 1H), 2.20 - 2.01 (m, 2H), 1.70 (dd, 7= 14.1, 11.7 Hz, 1H), 1.26 (d, 7= 6.4 Hz, 3H). LCMS m/z 325.01 [M+H]⁺.
Compound 73 oOlV^c· pXp-S H II ^NpT il	S24; H 5 MT	Compound 20	Ή NMR (400 MHz, DMSO7₆) δ 12.82 (s, 1H), 9.25 (d,7= 10.7 Hz, 1H), 8.72 (d, 7= 11.3 Hz, 1H), 6.94 (s, 1H), 6.17 (s, 1H), 4.47 (t, 7= 11.1 Hz, 1H), 3.94 (d, 7= 5.7 Hz, 2H), 3.623.51 (m, obscured by water, 1H), 2.61 (t, 7= 5.4 Hz, 2H), 2.41 (d, 7= 14.5 Hz, 1H), 2.24 (s, 4H), 2.10 (t, 7= 13.6 Hz, 1H), 1.86-1.77 (m, 1H), 1.27

203

Product	Starting Material and Aldéhyde	Method	Ή NMR; LCMS m/z [M+Hf
			(d, J= 6.4 Hz, 3H). LCMS m/z 338.17 [M+H]⁺.
Compound 74 (Ocv-ci AA S / λ ' H \ A^N	S24; 0 H <-nT / )^^N0^	Compound 20	Ή NMR (400 MHz, DMSOdô) δ 9.11 (d, J =10.8 Hz, 1H), 8.67 (s, 1H), 7.32 (s, 1H), 6.94 (s, 1H), 4.76 (d, J = 1.6 Hz, 2H), 4.45 (s, 1H), 4.01 (p, J= 4.7, 4.1 Hz, 4H), 3.93 (t, J= 5.5 Hz, 2H), 3.50 (s, 1H), 2.60 (t, J=5.3 Hz, 2H), 2.39-2.13 (m, 3H), 1.80 (dd,J= 14.4, 12.2 Hz, 1H), 1.24 (d, J =6.5 Hz, 3H). LCMS m/z 380.16 [M+H]⁺.
Compound 75 ₀O>^ciζ><Α O H N^hT 1	S25; 0 JJ fï A A 1	Compound 21	Ή NMR (400 MHz, DMSOdd) δ 8.93 (s, 1H), 8.61 (s, 1H), 8.49 (s, 2H), 6.94 (s, 1H), 4.45 (t, .7= 11.1 Hz, 1H), 3.94 (t, J= 5.5 Hz, 2H), 3.65 - 3.55 (m, 1H), 3.13 (s, 6H), 2.60 (t, J= 5.3 Hz, 2H), 2.41 -2.31 (m, 1H), 2.26 (d, .7= 13.8 Hz, 2H), 1.84 (t, J= 13.3 Hz, 1H), 1.25 (d, J= 6.4 Hz, 3H). LCMS m/z 379.17 [M+H]⁺.
Compound 76 oOO^^ciAç s ,\|\k A A, ΐ 1] H	S25; 0 A^J	Compound 21¹⁰	LCMS m/z 366.12 [M+H]⁺.
Compound 77 <OlV^cizC⁵ /A^n ^h	S25; 0 H UCzN	Compound 19	Ή NMR (400 MHz, DMSOJô) δ 9.30 (s, 1H), 8.87 (s, 1H), 8.53 (d, .7=2.1 Hz, 1H), 7.73 (d, .7= 8.0 Hz, 1H), 7.49 (d, J= 8.0 Hz, 1H), 6.94 (s, 1H), 4.64 (t, 1H), 3.99 (h, J =6.2 Hz, 2H), 3.65 - 3.56 (m, 1H), 2.62 (t, J =5.4 Hz, 2H), 2.43 (d, J= 14.4 Hz, 1H), 2.34 (s, 3H), 2.26 (d, J= 14.5 Hz, 1H), 2.081.98 (m, 1H), 1.96- 1.87 (m, 1H), 1.31 (d, J= 6.5 Hz, 3H). LCMS m/z 349.23 [M+H]⁺.

204

Product	Starting Material and Aldéhyde	Method	‘H NMR; LCMS m/z [M+Hf
Compound 78 _oOl>-^ci F II q_x L 1] h h₂n'%	S25; F 0 h₂n	Compound 21	Ή NMR (400 MHz, DMSOd₆) δ 9.37 (s, 1H), 8.92 (s, 1H), 7.91 (t, J =7.6 Hz, 1H), 7.79 (d, 7=8.5 Hz, 1H), 7.69 (d, J= 9.4 Hz, 1H), 7.62 (s, 2H), 6.94 (s, 1H), 4.82 (t, J=11.5 Hz, 1H), 3.98 (t, 7= 5.5 Hz, 2H), 3.72 (s, 1H), 2.62 (t, 7= 5.4 Hz, 2H), 2.42-2.18 (m, 3H), 1.93 (t, 7 = 13.3 Hz, 1H), 1.30 (d, J= 6.4 Hz, 3H). LCMS m/z 431.01 [M+H]⁺.
Compound 79 oCO-c JxJ^S °U ^H	S25; O H °\2n	Compound 21¹¹	Ή NMR (400 MHz, DMSOd₆) δ 9.28 (s, 1H), 8.85 (s, 1H), 8.57 (s, 1H), 8.31 (s, 1H), 6.95 (s, 1H), 4.60 (t, 7 = 11.1 Hz, 1H), 3.94 (t, 7= 5.3 Hz, 2H), 3.58 (s, 1H), 2.60 (t, 7= 5.5 Hz, 2H), 2.44-2.15 (m, 3H), 1.89- 1.78 (m, 1H), 1.26 (d,7 = 6.4 Hz, 3H). LCMS m/z 325.12 [M+H]⁺.
Compound 80 oOl^^ci r^'V^N^ Γ JJ H crV 1	S25; 0 H ifj\| CT^N 1	Compound 21	*H NMR (400 MHz, DMSO7_ά) δ 8.94 (s, 1H), 8.56 (s, 1H), 7.96 (d, 7=2.6 Hz, 1H), 7.63 (dt, 7=9.5, 4.7 Hz, 1H), 6.94 (s, 1H), 6.46 (d, 7=9.4 Hz, 1H), 4.46 - 4.28 (m, 1H), 3.95 3.91 (m, 2H), 3.65 - 3.49 (m, 1H), 3.43 (s, 3H), 2.62-2.57 (m, 2H), 2.33 -2.17 (m, 3H), 1.96- 1.70 (m, 1H), 1.25 (d,7 = 6.5 Hz, 3H). LCMS m/z 365.13 [M+H]⁺.
Compound 81 f^TVe^{1 s/} LJ H	S25; 0 H Q]	Compound 19	Ή NMR (400 MHz, DMSOde) δ 9.11 (s, 1H), 8.59 (s, 1H), 7.63 - 7.54 (m, 2H), 7.52 - 7.41 (m, 3H), 6.94 (s, 1H), 4.54 (t, J = 11.3 Hz, 1H), 3.97 (t,7=5.4 Hz, 2H), 3.65 (s, 1H),2.642.58 (m, 2H), 2.27 (dt,7= 20.9, 13.8 Hz, 3H), 1.95-1.86 (m, 1H), 1.28 (d, 7= 6.5 Hz, 3H). LCMS m/z 334.19 [M+H]⁺.

205

Product	Starting Material and Aldéhyde	Method	’HNMR; LCMS m/z [M+Ilf
Compound 82 oOlV^ci ^h	S25; O JJ jl J	Compound 19	*H NMR (400 MHz, DMSOde) δ 9.07 (s, 1H), 8.88 (s, 2H), 8.78 (s, 1H), 6.95 (s, 1H), 4.71 -4.61 (m, 1H), 3.96 (t, 7=5.5 Hz, 2H), 2.98 -2.89 (m, 1H), 2.65 (s, 3H), 2.61 (dd,7=8.5, 4.8 Hz, 2H), 2.38 (d, 7= 8.1 Hz, 2H), 2.28 (d, 7 = 14.5 Hz, 1H), 1.93 - 1.83 (m, 1H), 1.28 (d, J= 6.6 Hz, 3H). LCMS m/z 350.22 [M+H]⁺.
Compound 83 <OlA^ci A>N ^H	S25; 0 H On	Compound 19	Ή NMR (400 MHz, DMSOd₆) δ 9.37 (s, 1H), 8.92 (s, 1H), 8.69 (d, 7=4.9 Hz, 1H), 7.92 (td, 7=7.8, 1.9 Hz, 1H), 7.60 (d, 7=7.9 Hz, 1H), 7.48 (dd,7 = 7.4, 4.8 Hz, 1H), 6.94 (s, 1H), 4.70 (s, 1H), 4.00 (dp, 7= 11.6, 5.9, 5.5 Hz, 2H), 3.60 (s, 1 H), 2.63 (t, 7=5.3 Hz, 2H), 2.46 (d, 7= 14.3 Hz, 1H), 2.26 (d, 7= 14.3 Hz, 1H), 2.09 1.99 (m, 1H), 1.97- 1.89 (m, 1H), 1.32 (d, 7= 6.5 Hz, 3H). LCMS m/z 335.19 [M+H]⁺.
Compound 84 cOCVci ^s _/N=<v pr ° W^NH	S24; 0 H N=(7 /^0HW^NH	Compound 20	’H NMR (400 MHz, DMSOd₆} δ 11.42 (s, 1H), 9.29 (d,7= 10.7 Hz, 1H), 8.93 (d,7= 11.3 Hz, 1H), 7.75 (dd, 7= 8.8, 5.7 Hz, 1H), 6.96 (s, 1H), 6.65 6.57 (m, 2H), 4.74 (t, 7= 11.3 Hz, 1H), 3.98 (t, 7= 5.3 Hz, 2H), 3.84 (s, 3H), 2.67 - 2.58 (m, 2H), 2.39 - 2.26 (m, 2H), 1.94- 1.80 (m, 1H), 1.30 (d, 7 = 6.5 Hz, 3H). Note: 1H appears to be hidden under DMSO and 1H hidden under water. LCMS m/z 404.14 [M+H]⁺.
Compound 85 oOlV^ciAx ^S Νχ/kA <^zjr il Hhr	S24; 0 <^ZJ	Compound 20¹·⁶	’H NMR (400 MHz, DMSO7₆) δ 7.56 (s, 1H), 6.94 (s, 1H), 6.88 (s, 1H), 4.13-4.05 (m, 1H), 3.89 (t, 7= 5.4 Hz, 2H), 3.64 - 3.29 (m, under water, 1H), 3.22-3.11 (m, 1H), 2.57 (t, 7= 5.2 Hz, 2H), 2.20 (d,7= 13.6 Hz, 1H), 2.02 (d, 7= 13.6

206

Product	Starting Material and Aldéhyde	Method	^JHNMR; LCMS m/z [M+HJ'
			Hz, 1H), 1.72 (t, J= 12.6 Hz, 1H), 1.37 (t, J= 12.5 Hz, 1H), 1.06 (d, J =6.3 Hz, 3H). LCMS m/z 324.17 [M+H]⁺.
Compound 86 oOiX^CIAA⁵ A ,N ^H HO N	S25; 0 Hcr>r^N	Compound 20	H NMR (400 MHz, DMSOd₆) δ 13.37 (s, 1H), 9.36 (d, J= 8.7 Hz, 1H), 8.90 - 8.72 (m, 1H), 7.60 (d, J =9.9 Hz, 1H), 7.03 - 6.96 (m, 1H), 6.95 (s, 1H), 4.58 (t, 7= 11.1 Hz, 1H), 3.97 (tp, 7= 11.8, 5.6 Hz, 2H), 3.55 (s, 1H), 2.61 (t,7=5.4 Hz, 2H), 2.39- 1.75 (m, 3H), 1.28 (d, 7= 6.5 Hz, 3H). Note: 1H is hidden under DMSO peak. LCMS m/z 352.17 [M+H]⁺.
Compound 87 (OlV^ci η ^HN'°	S25; 0 I] N-°	Compound 21	‘H NMR (400 MHz, DMSO7₆)δ9.51 (s, 1H), 9.11 (s, 1H), 6.95 (s, 1H), 6.52 (s, 1H), 4.78 (s, 1H), 3.94 (t, 7= 5.4 Hz, 2H), 3.62 (s, 1H), 2.60 (t, 2H), 2.57-2.47 (m, hidden under DMSO, 1H), 2.24 (d,7= 14.1 Hz, 1H), 2.16 (t, 7= 13.5 Hz, 1H), 2.05 (tt, 7=8.8, 4.1 Hz, 1H), 1.80 (t, 7= 13.4 Hz, 1H), 1.26 (d, 7= 6.5 Hz, 3H), 1.101.01 (m, 2H), 0.80-0.70 (m, 2H). LCMS m/z 365.13 [M+H]⁺.
Compound 88 oO^X^ci AJ N_>rJX.^U, Np PI N J	S25; H 5 J N J W	Compound 21	^rH NMR (400 MHz, DMSO7₆) δ 13.86 (s, 1 H), 9.38 (d,7= 10.4 Hz, 1H), 8.87 (d,7= 12.1 Hz, 1H), 8.64 (d, 7=4.9 Hz, 1H), 7.92 (t, 7= 7.8 Hz, 1H), 7.83 (d, 7= 7.9 Hz, 1H), 7.39 (t, 7=6.3 Hz, 1H), 7.07 (s, 1H), 6.95 (s, 1H), 4.60 (d,7= 10.2 Hz, 1H), 4.02-3.94 (m, 2H), 3.60 (s, 1H), 2.65-2.60 (m, 2H), 2.37 - 2.07 (m, 2H), 1.90- 1.79 (m, 1H), 1.29 (d,7 = 6.4 Hz, 3H), 1.25 (d,7=6.2 Hz, 1H). LCMS m/z 380.16 [M+H]⁺.

207

Product	Starting Material and Aldéhyde	Method	^!HNMR; LCMS m/z [M+Ilf
Compound 89 _oO>^c' το^s ï JJ H 1	S25; 0 H îj 1	Compound 21	*H NMR (400 MHz, DMSO7₆) δ 8.97 (s, 1H), 8.55 (s, 1H), 8.20 (d, 7=2.4 Hz, 1H), 7.76 (d, 7= 9.0 Hz, 1H), 6.94 (s, 1H), 6.79 (d, 7=8.9 Hz, 1H), 4.47 (t, 7= 11.1 Hz, 1H), 3.95 (t, 7=5.4 Hz, 2H), 3.61 (s, 1H), 3.06 (s, 6H), 2.60 (t, 7= 5.5 Hz, 2H), 2.35-2.17 (m, 3H), 1.90-1.81 (m, 1H), 1.26 (d, 7= 6.6 Hz, 3H). LCMS m/z 378.17 [M+H]⁺.
Compound 90 _O00^cikk'⁵ <Th lé —0	S25; 0 H lé —0	Compound 21	*H NMR (400 MHz, DMSO7ô) δ 8.96 (s, 1H), 8.54 (s, 1H), 7.89 (s, 1H), 7.62 (s, 1H), 6.94 (s, 1H), 4.50 (t, 7= 11.0 Hz, 1H), 4.25 (t, 7=5.2 Hz, 2H), 3.93 (t, 7 = 5.4 Hz, 2H), 3.67 (t, 7=5.2 Hz, 2H), 3.55 (s, 1H), 3.22 (s, 3H), 2.59 (s, 2H), 2.36 (d, 7= 14.3 Hz, 1H), 2.28 2.11 (m, 2H), 1.79 (t, 7= 13.3 Hz, 1H), 1.25 (d, 7=6.6 Hz, 3H). LCMS m/z 382.15 [M+H]⁺.
Compound 91 <OlV^ciz\ ^s _zck Τ_ΜΤ, M H	S25; O _ZO^J XJ	Compound 21¹²	‘H NMR (400 MHz, DMSOd₆) δ 9.55 (s, lH),9.14(s, 1H), 6.95 (s, 1H), 6.70 (s, 1H), 4.83 (s, 1H), 3.95 (t, 7= 5.5 Hz, 2H), 3.64 (s, 1H), 2.66 (q,7 = 7.6 Hz, 3H), 2.61 (s, 2H), 2.31 -2.13 (m, 2H), 1.82 (t, 7= 13.2 Hz, 1H), 1.27 (d, 7=6.4 Hz, 3H), 1.19 (t, 7= 7.5 Hz, 3H). LCMS m/z 353.16 [M+H]⁺.
Compound 92 (OC^ci H 1 H	S25; 0 N-V H \	Compound 19¹³	Ή NMR (400 MHz, DMSOd₆) δ 9.56 (s, 1H), 9.33 (dd,7= 4.9, 1.6 Hz, 1H), 9.27 (d,7= 12.3 Hz, 1H), 7.94 (dd,7=8.7, 1.6 Hz, 1H), 7.85 (dd,7=8.5, 4.9 Hz, 1H), 6.96 (s, 1H), 4.94 (d, 7= 11.6 Hz, 1H), 4.02 (tt, 7 = 11.9, 6.3 Hz, 2H), 3.63 (s, 1H), 2.64 (t, 7= 5.4 Hz, 3H), 2.30 (d, 7= 14.6 Hz, 1H), 2.07 (t, 7= 13.4 Hz, 1H), 2.00-1.89 (m, 1H), 1.35 (d, 7= 6.5 Hz,

208

Product	Starting Material and Aldéhyde	Method	’H NMR; LCMS m/z [M+Ilf
			3H). LCMS m/z 336.14 [M+H]⁺.
Compound 93 oOC^^ci	S25; O σ	Compound 19	‘H NMR (400 MHz, DMSOd₆) δ 9.43 (d, 7=2.3 Hz, 1H), 9.36 (d, 7= 5.4 Hz, 1H), 9.13 (s, 1H), 8.79 (s, 1H), 7.88 (dd, 7=5.4, 2.4 Hz, 1H), 6.95 (s, 1H), 4.75-4.65 (m, 1H), 3.98 (t, 7= 5.9 Hz, 2H), 3.68 - 3.65 (m, 1H), 2.63 (t,7=4.7 Hz, 2H), 2.45 (d, 7= 15.0 Hz, under DMSO, 1H), 2.29 (d, J= 15.0 Hz, 1H), 2.22 (t, 7= 13.7 Hz, 1H), 1.87 (t, 7= 13.4 Hz, 1H), 1.30 (d, 7= 6.5 Hz, 3H). LCMS m/z 336.18 [M+H]⁺.
Compound 94 <Q>^aXjAA	S25; 0	Compound 21	*H NMR (400 MHz, DMSO7₆) δ 9.29 (d, J= 35.4 Hz, 1H), 8.95 (s, 1H), 7.20 (s, 1H), 6.94 (s, 1H), 4.72 (t, 7= 11.1 Hz, 1H), 3.93 (t, 7= 5.5 Hz, 2H), 3.60 (s, 1H), 2.60 (q,7=4.7 Hz, 2H), 2.46 - 2.38 (m, 4H), 2.28-2.16 (m, 2H), 1.82 (dd,7 = 14.4, 12.3 Hz, 1H), 1.25 (d, 7 = 6.6 Hz, 3H). LCMS m/z 339.16 [M+H]⁺.
Compound 95 _oOlV^{ci S} A J ^HHN N OH	S24; 0 JJ Λ J HN N OH S22	Compound ig3,4,14	Ή NMR (300 MHz, MethanolJ₄) δ 8.40 (s, 2H), 8.23 (s, 3H), 6.75 (s, 1H), 4.57 (dd,7= 12.5, 3.1 Hz, 1H), 3.99 (t, 7= 5.5 Hz, 2H), 3.80 (ddd, 7=12.2, 6.6, 2.8 Hz, 1H), 3.66 (s, 2H), 2.69 - 2.63 (m, 2H), 2.45 - 2.20 (m, 3H), 1.81 (dd, 7= 14.7, 12.2 Hz, 1H), 1.36 (d, 7= 6.6 Hz, 9H). LCMS m/z 423.19 [M+H]⁺.
Compound 96 <OO^^cl \ m H 0-N	S25; 0 H Z» q-N	Compound 21	’H NMR (400 MHz, DMSO7ô) δ 9.64 (s, 1H), 9.17 (s, 1H), 9.06 (d, 7=1.7 Hz, 1H), 6.95 (s, 1H), 6.86 (d,7= 1.7 Hz, 1H), 4.81 (s, 1H), 3.98 (q,7= 5.4 Hz, 2H), 3.62 (s, 1H), 2.62 (t, 7= 5.7 Hz, 2H), 2.61-2.53 (m, 1H), 2.28 (d, 7= 14.4 Hz, 1H), 2.10 (t, 7= 13.5 Hz, 1H), 1.90- 1.79 (m, 1H), 1.30 (d, 7

209

Product	Starting Material and Aldéhyde	Method	‘HNMS; LCMS m/z [M+H]⁺
			= 6.5 Hz, 3H). LCMS m/z 325.12 [M+H]⁺.
Compound 97 (OCV^C\| Aj h °=S^/ 0	S25; 0 IJ n'a °=S^/ u 0	Compound 20	Ή NMR (300 MHz, MethanolJ₄) δ 7.97 (s, 1H), 7.73 (s, 1H), 6.76 (s, 1H), 5.27 (p, 7=7.2 Hz, 1H), 4.71 (dd, 7= 12.5, 2.9 Hz, 1H), 3.99 (t, 7= 5.5 Hz, 2H), 3.78 (s, 1 H), 3.65 (dd,7= 13.8, 8.1Hz, 1H), 3.56-3.39 (m, 2H), 3.23 (dd, 7= 13.4, 7.7 Hz, 1H), 2.69 (dt, 7= 15.4,6.1 Hz, 4H), 2.58 - 2.44 (m, 1H), 2.38 (dt, 7= 14.7 Hz, 1H), 2.19 (dd, J= 14.7, 12.7 Hz, 1H), 1.79 (dd, 7= 14.7, 12.2 Hz, 1H), 1.37 (d, 7= 6.6 Hz, 3H). LCMS m/z 442.11 [M+H]⁺.
Compound 98 oOlV^c\|AA⁵ ,nA\A •--M N NJ H	S25; 0 A ~~ⁿCj	Compound 21	‘H NMR (300 MHz, Methanol70 δ 7.97 (s, 1H), 7.73 (s, 1H), 6.76 (s, 1H), 5.27 (p,7=7.2 Hz, 1H), 4.71 (dd, 7= 12.5, 2.9 Hz, 1H), 3.99 (t, 7= 5.5 Hz, 2H), 3.78 (s, 1 H), 3.65 (dd,7= 13.8, 8.1 Hz, 1H), 3.56-3.39 (m, 2H), 3.23 (dd, 7= 13.4, 7.7 Hz, 1H), 2.69 (dt, 7=15.4, 6.1 Hz, 4H), 2.58 - 2.44 (m, 1H), 2.38 (dt, 7= 14.7 Hz, 1H), 2.19 (dd, 7= 14.7, 12.7 Hz, 1H), 1.79 (dd, 7=14.7, 12.2 Hz, 1H), 1.37 (d, 7= 6.6 Hz, 3H). LCMS m/z 352.17 [M+H]⁺.
Compound 99 oOO^^ci H(X^A_KA L h	S25; 0 Tj	Compound 21	Ή NMR (400 MHz, DMSOd₆) δ 9.22 (s, 1H), 8.97 (d,7 = 10.5 Hz, 1H), 8.41 (s, 1H), 7.01 - 6.90 (m, 4H), 4.38 (t,7= 10.7 Hz, 1H), 3.95 (t, 7=5.5 Hz, 2H), 3.78 (s, 3H), 3.60 (s, 1H), 2.60 (t, 7= 5.4 Hz, 2H), 2.22 (t, 7= 16.8 Hz, 3H), 1.86 (t, 7= 13.4 Hz, 1H), 1.26 (d,7=6.5 Hz, 3H). LCMS m/z 380.16 [M+H]⁺.

210

Product	Starting Material and Aldéhyde	Method	^!HNMR; LCMS m/z [M+Hf
Compound 100 Nfj) fl A 0	S25; 0 H lA	Compound 21	Ή NMR (300 MHz, MethanolJ₄) δ 6.54 (s, 1H), 6.30 (s, 1H), 5.38 (s, 1H), 3.34 (dd, 7=12.7, 3.0 Hz, 1H), 3.13 -2.99 (m, 1H), 2.73 - 2.62 (m, 2H), 2.62 (t, 7= 5.5 Hz, 2H), 2.39 (d, 7= 8.1 Hz, 1H), 2.19 (td, 7= 11.6, 11.2, 4.4 Hz, 2H), 1.31-1.26 (m, 2H), 1.14 (dt, 7= 14.7, 2.7 Hz, 1H), 0.99 (dt, 7= 14.5, 2.7 Hz, 1H), 0.83 (dd, 7=14.7, 12.7 Hz, 1H), 0.67 (td, 7= 9.7, 8.6,3.9 Hz, 4H), 0.42 (dd,7= 14.7, 12.2 Hz, 1H), -0.01 (d,7 = 6.6 Hz, 3H). LCMS m/z 408.11 [M+H]⁺.
Compound 101 oOlV^ciz<C^s nAA M ^H	S24; 0 H Υ'Ύ	Compound 20	’H NMR (400 MHz, DMSOdè) δ 9.42 (s, 1H), 9.01 (d,7= 10.9 Hz, 1H), 8.89 (d,7=1.5 Hz, 1H), 8.79 - 8.72 (m, 2H), 6.95 (s, 1H), 4.86 (t, 7= 11.3 Hz, 1H), 4.00 (tt, 7= 11.4, 5.7 Hz, 2H), 3.64 (s, 1H), 2.63 (t,7 = 5.4 Hz, 2H), 2.58-2.51 (m, 1H), 2.27 (d, 7= 14.6 Hz, 1H), 2.15-2.05 (m, 1H), 1.98 - 1.88 (m, 1H), 1.31 (d, 7=6.5 Hz, 3H). LCMS m/z 336.14 [M+H]⁺.
Compound 102 _oOCV-^ciAx ^s JL J ^H HO^y f°	S25; 0 H Ηο^γ r°	Compound 21	‘H NMR (400 MHz, DMSOd₆) δ 9.23 (s, 1H), 8.98 (d,7= 10.1 Hz, 1H), 8.46 (s, 1H), 7.15 (d, 7=2.1 Hz, 1H), 6.93 (q,7= 2.6, 2.2 Hz, 2H), 6.82 (d,7= 8.2 Hz, 1H), 4.49 - 4.33 (m, 1H), 4.04 (q, 7= 7.0 Hz, 2H), 3.94 (t, 7= 5.5 Hz, 2H), 3.59 (s, 1H), 2.60 (t, 7=5.4 Hz, 2H), 2.25 (t, 3H), 1.87 (t,7= 13.3 Hz, 1H), 1.34 (t, 7= 7.0 Hz, 3H), 1.27 (d, 7= 6.5 Hz, 3H). LCMS m/z 394.16 [M+H]⁺.

211

Product	Starting Material and Aldéhyde	Method	’lLVMR; LCMS m/z [M+Hf
Compound 103 tOO^^ci H HO \ y^NHO—/	S24; 0 Π Lj TBSO \__ TBScA S21	Compound igl,3,4,5	Ή NMR (300 MHz, Chloroformé) δ 7.48 (s, 1H), 7.41 (s, 1H), 6.58 (s, 1H), 4.23 - 4.14 (m, 3H), 3.94 (t, 7= 5.4 Hz, 2H), 3.49 (d, 7= 11.3 Hz, 2H), 3.32 (d, 7= 11.5 Hz, 2H), 3.31 -3.18 (m, 1H), 2.61 (t,7 = 5.4 Hz, 2H), 2.22 (dt,7= 13.7, 2.5 Hz, 1H), 2.07 (dd, 7 = 13.7, 2.6 Hz, 1H), 1.75 - 1.64 (m, 1H), 1.41 (dd, 7= 13.8, 11.3 Hz, 1H), 1.13 (d, 7=6.4 Hz, 3H), 0.79 (s, 3H). LCMS m/z 426.13 [M+H]⁺.
Compound 104 CA ^s O. ¹ H VN ⁿ	S25; O H CD/ N	Compound 20	Ή NMR (400 MHz, DMSO7₆) δ 9.27 - 9.15 (m, 1H), 8.81 (d,7= 11.9 Hz, 1H), 8.14 (s, 1H), 6.94 (s, 1H), 4.51 (t,7= 11.1 Hz, 1H), 3.93 (t,7=5.4 Hz, 2H), 3.60-3.51 (m, 1H), 2.60 (t, 7= 5.4 Hz, 2H), 2.46 (s, 3H), 2.38 - 2.08 (m, 3H), 1.81 (dd, 7= 14.5, 12.2 Hz, 1H), 1.25 (d, 7= 6.5 Hz, 3H). LCMS m/z 339.16 [M+H]⁺.
Compound 105 cOCVci A nA ή ' ï L J ^H Cj^N	S25; 0 ïï A nJ/ 0^	Compound 21	*H NMR (300 MHz, MethanolJ₄) δ 9.03 (s, 2H), 7.67 (dd, 7= 7.6, 1.8 Hz, 1H), 7.55-7.47 (m, 1H), 7.17 (d, 7= 8.4 Hz, 1H), 7.08 (dd, 7=8.0, 7.0 Hz, 1H), 6.78 (s, 1H), 4.92 (dd,7= 12.8, 2.9 Hz, 1H), 4.05 (t, 7= 5.5 Hz, 2H), 4.00 - 3.86 (m, 1 H), 3.83 (s, 3H), 2.69 (t,7= 5.5 Hz, 2H), 2.61 (d,7= 14.5 Hz, 1H), 2.51 -2.30 (m, 2H), 1.90 (dd, 7= 14.8, 12.2 Hz, 1H), 1.43 (d, 7= 6.6 Hz, 3H). LCMS m/z 442.11 [M+H]⁺.
Compound 106 (OcV^Gi/A^s / N °A ^H o—/ /	S25; O O J CA !	Compound 21	Ή NMR (400 MHz, DMSO7β) δ 9.60 (s, 1H), 9.14 (s, 1H), 6.95 (d, 7=1.2 Hz, 1H), 6.78 (s, 1H), 4.77 (t, 7= 11.2 Hz, 1H), 4.59 (s, 2H), 3.97 (hept, 7 = 6.1, 5.7 Hz, 2H), 3.62 (s, lH),3.32(s, 3H), 2.62 (t, 7= 5.6 Hz, 2H), 2.60 - 2.53 (m, 1H), 2.27 (d, 7= 14.0 Hz, 1H),

212

Product	Starting Material and Aldéhyde	Method	‘H NMR; LCMS m/z [M+Hf
			2.09 (t, J = 13.5 Hz, 1H), 1.901.78 (m, 1H), 1.29 (d, J=6.4 Hz, 3H). LCMS m/z 369.10 [M+H]⁺.
Compound 107 cOCVci ζχγ-s Y \|f N LJ H	S25; 0 HO^xxJ TJ ^/Xo^/x/	Compound 21¹·⁶	Ή NMR (400 MHz, DMSOJ₆) δ 8.79 (s, 1H), 6.85 (s, 2H), 6.84-6.69 (m, 2H), 3.97 (q, J = 6.9 Hz, 2H), 3.90 (t, J =5.4 Hz, 3H), 3.11 (s, 1H), 2.592.55 (m, 2H), 1.99 (d, J = 13.3 Hz, 2H), 1.56-1.19 (m, 2H), 1.30 (t, J =6.9 Hz, 3H), 1.03 (d, J= 6.2 Hz, 3H). LCMS m/z 394.16 [M+H]⁺.
Compound 108 oOtV^ci mm Il 1 h HCiy	S25; O H JLJ hcM/	Compound 20	‘H NMR (400 MHz, DMSOd₆) δ 9.65 (s, 1 H), 8.92 (d, J= 10.7 Hz, 1H), 8.40 (s, 1H), 7.27 (d, .7=2.3 Hz, 1H), 7.17 (d,J= 9.1 Hz, 1H), 6.93 (s, 1H), 6.81 (d, .7= 8.2 Hz, 1H), 4.42 - 4.30 (m, 1H), 3.94 (t, .7= 5.5 Hz, 2H), 3.65 -3.55 (m, 1H), 2.59 (t, .7= 5.9 Hz, 2H), 2.30-2.18 (m, 3H), 2.14 (s, 3H), 1.86 (t, J = 13.3 Hz, 1H), 1.25 (d, .7=6.5 Hz, 3H). LCMS m/z 364.18 [M+H]⁺.
Compound 109 (OlV^ciSçS L 1] h H₂NX)	S25; 0 1) H₂N X)	Compound 21	‘H NMR (400 MHz, DMSOîZ₆)ô9.12(s, 1H), 8.62 (d, J= 12.3 Hz, 1H), 8.09 (d,J=7.6 Hz, 2H), 7.93 (d, J =1.6 Hz, 1H), 7.71 (d, .7=7.7 Hz, 1H), 7.54 (dd, J= 13.4, 5.7 Hz, 2H), 6.94 (s, 1H), 4.60 (t, J = 11.0 Hz, 1H), 3.97 (t, .7=5.4 Hz, 2H), 3.70-3.62 (m, 1H), 2.63 2.59 (m, 2H), 2.30 (dt, J = 26.0, 12.8 Hz, 3H), 1.91 (t, J= 13.3 Hz, 1H), 1.28 (d, J =6.5 Hz, 3H). LCMS m/z 377.09 [M+H]⁺.
Compound 110 (OC^^{ci s}	S24; 0 ii Ύ	Compound 20	Ή NMR (400 MHz, DMSOd₆) δ 9.52 (d, J= 10.5 Hz, 1H), 8.96 (d, J= 11.3 Hz, 1H), 8.70 (d, J= 5.0 Hz, 1H), 7.36 (d, J= 5.2 Hz, 1H), 6.95 (s, 1H), 4.69 (t, J= 11.3 Hz, 1H), 4.11-3.92 (m, 5H), 3.63 -3.54 (m, 1H),

213

Product	Starting Material and Aldéhyde	Method	'HNMH; LCMS m/z [M+Hp
			2.63 (t, 7=5.4 Hz, 2H), 2.58 2.19 (m, 2H), 1.94 (dt,7 = 27.4, 13.5 Hz, 2H), 1.33 (d,7= 6.5 Hz, 3H). LCMS m/z 366.16 [M+H]⁺.
Compound 111 /“x ^s N' J 9	S25; 0 N? J	Compound 21	Ή NMR (400 MHz, DMSOde) δ 9.38 (s, 1H), 8.92 (d,7= 11.4 Hz, 1H), 8.21 (d,7= 1.8 Hz, 1H), 6.95 (s, 1H), 5.07 (s, 1H), 4.72 (t, 7= 11.1 Hz, 1H), 4.42 (dd, 7= 13.9, 3.9 Hz, 1H), 4.29 (dd, 7= 13.8,7.1 Hz, 1H), 3.96 (t, 7= 5.2 Hz, 2H), 3.74 (s, 1H), 3.61 (s, 1H), 2.61 (s, 2H), 2.45 (s, 1H), 2.25 (t, 7 = 13 .4 Hz, 2H), 1.92- 1.76 (m, 1H), 1.44- 1.30 (m, 2H), 1.27 (d, 7= 6.5 Hz, 3H), 0.91 (t,7= 7.4 Hz, 3H). LCMS m/z 397.14 [M+H]⁺.
Compound 112 (OlVci çrOOH	S25; 0 H ΙΙΊ OH	Compound 21	*H NMR (400 MHz, DMSO7,) δ 11.77 (s, 1H), 9.23 (s, 1H), 8.68 (s, 1H), 7.46 (d,7= 6.9 Hz, 1H), 6.93 (s, 1H), 6.50 (d, 7= 1.8 Hz, 1H), 6.32 (d,7= 6.7 Hz, 1H), 4.36 (t, 7= 11.5 Hz, 1H), 3.98 -3.93 (m, 2H), 3.59 (s, 1H), 2.62-2.58 (m, 2H), 2.37-2.19 (m, 2H), 2.05 (t, 7= 13.4 Hz, 1H), 1.86 (dd,7 = 14.3, 12.3 Hz, 1H), 1.28 (d,7 = 6.6 Hz, 3H). LCMS m/z 351.13 [M+H]⁺.
Compound 113 _οΟίΑ^{α s} H O ^H	S25; ^H U o AA	Compound 21	*H NMR (300 MHz, DMSOdè) δ 10.07 (s, 1H), 9.04 (s, 1H), 8.51 (s, 1H), 7.89 (s, 1H), 7.48 (d, 7=8.1 Hz, 1H), 7.39 (t, 7= 7.8 Hz, 1H), 7.25 (d,7= 7.6 Hz, 1H), 6.94 (s, 1H), 4.47 (t, 7= 11.0 Hz, 1H), 3.96 (t, 7= 5.4 Hz, 2H), 3.88 - 3.50 (m, under water, 1 H), 2.61 (t, 7= 5.4 Hz, 2H), 2.22 (dd, 7= 24.7, 11.8 Hz, 3H), 2.06 (s, 3H), 1.96 -1.82 (m, 1H), 1.27 (d,7=6.5 Hz, 3H). LCMS m/z 391.13 [M+H]⁺.

214

Product	Starting Material and Aldéhyde	Method	'H NMR; LCMS m/z [M+Hf
Compound 114 oOlX~^ciγχγ s ^HO H o-'	S25; 0 ⁿX HN T qA	Compound 21	’H NMR (400 MHz, DMSOde) δ 12.92 (s, 1H), 9.24 (s, 1H), 8.74 (s, 1H), 6.94 (s, 1H), 6.25 (s, 1H), 4.49 (t, J= 11.4 Hz, 1H), 3.95 (t, 2H), 3.90 (d, J = 9.1 Hz, 2H), 3.54 (s, 1H), 3.46-3.41 (m, 2H), 2.90 (d, J = 12.3 Hz, 1H), 2.61 (d, J =5.6 Hz, 2H), 2.44 (d, J= 14.6 Hz, 1H), 2.23 (d, 7= 14.5 Hz, 1H), 2.17-2.07 (m, 1H), 1.80 (d, J = 13.2 Hz, 3H), 1.60 (tt, J = 12.3,6.2 Hz, 2H), 1.26 (d, J= 6.4 Hz, 3H). LCMS m/z 408.11 [M+H]⁺.
Compound 115 (OC^^ci~>çs O H % .^N A >, A, b IL^J ^H	S25; ° H U Ai ΥΑΧ ⁰ AA	Compound 21	Ή NMR (400 MHz, DMSOd₆) δ 9.93 (s, 1H), 9.09 (d, J= 10.2 Hz, 1H), 8.59 (s, 1H), 7.44 (t, 7= 7.8 Hz, 1H), 7.37-7.30 (m, 2H), 7.23 (d, J =8.1 Hz, 1H), 6.94 (s, 1H), 4.51 (t, J= 11.1 Hz, 1H), 3.96 (t, 7= 5.4 Hz, 2H), 3.66 (s, 1H), 3.03 (s, 3H), 2.61 (t, J =5.5 Hz, 2H), 2.26 (dt, 7=28.3,13.6 Hz, 3H), 1.90 (t, J= 13.3 Hz, 1H), 1.28 (d, 7= 6.5 Hz, 3H). LCMS m/z 427.08 [M+H]⁺.
Compound 116 oOA^c' γΧΑ M ^H	S24; 0 X T	Compound 20	*H NMR (400 MHz, DMSOde) δ 9.46 (s, 1H), 8.99 (d,7= 11.1 Hz, 1H), 8.79 (d,7=5.2 Hz, 1H), 7.54 (d, 7= 5.2 Hz, 1H), 6.95 (s, 1H), 4.69 (t, 7= 11.2 Hz, 1H), 4.00 (dp, 7= 17.5, 6.0 Hz, 2H), 3.58 (s, 1H), 2.71 (s, 3H), 2.63 (t,7=5.3 Hz, 2H), 2.25 (d, 7= 14.5 Hz, 1H), 2.42-2.11 (m, 1H), 1.95 (dt, 7=22.3, 13.5 Hz, 2H), 1.33 (d, 7= 6.5 Hz, 3H). LCMS m/z 350.18 [M+H]⁺.

215

Product	Starting Material and Aldéhyde	Method	'H NMR; LCMS m/z [M+Hf
Compound 117 ncx. N^J) H Ν-χ. w	S25; 0 κχ N -\ M	Compound 19	*H NMR (300 MHz, MethanolJ₄) δ 8.60 (d, .7=7.0 Hz, 1H), 8.19 (s, 1H), 7.88 (d, .7= 9.0 Hz, 1H), 7.40 (t, .7= 7.9 Hz, 1H), 7.02 (t, J= 6.8 Hz, 1H), 6.77 (s, 1H), 5.09 - 5.02 (m, 1H), 4.04 (t, J= 5.5 Hz, 2H), 3.93 (s, 1H), 2.68 (t, J =5.5 Hz, 2H), 2.58 -2.38 (m, 3H), 1.88 (dd, J= 14.7, 12.2 Hz, 1H), 1.39 (d, J =6.5 Hz, 3H). LCMS m/z 374.16 [M+H]⁺.
Compound 116 oOl^^cixC⁵ -—.m / N ^N\^J H	S25; 0 —n^Jj	Compound 20	Ή NMR (400 MHz, DMSOόΖ₆) δ 9.27 (d, J= 12.1 Hz, 1H), 8.77 (d, .7= 11.0 Hz, 1H), 7.77 (d, .7=2.3 Hz, 1H), 6.94 (s, 1H), 6.42 (dd, <7=10.9, 2.3 Hz, 1H), 4.52 (t,J= 11.1 Hz, 1H), 3.95 (t, .7= 5.4 Hz, 2H), 3.86 (s, 3H), 3.56 (s, 1H), 2.60 (t, J = 5.3 Hz, 2H), 2.34 (dd, J= 75.5, 14.6 Hz, 2H), 2.17 - 2.06 (m, 1H), 1.81 (dd, J= 14.5, 12.2 Hz, 1H), 1.26 (d, J =6.5 Hz, 3H). LCMS m/z 338.17 [M+H]⁺.
Compound 119 (OlV^cixx~^s ^HNÙ H F ^F	S25; 0 HN JJ ^FxC F ^F	Compound 21	‘H NMR (400 MHz, DMSOde) δ 13.96 (s, 1H), 9.29 (s, 1H), 9.05 (s, 1H), 6.96 (s, 1H), 6.95 (s, 1H), 4.73 (s, 1H), 3.96 (t, J= 5.3 Hz, 2H), 3.60 (s, 1H), 2.60 (d, J =5.9 Hz, 2H), 2.26 (d, J= 14.4 Hz, 2H), 1.83 (t, J= 13.2 Hz, 1H), 1.27 (d, J = 6.5 Hz, 3H). Note: 1H obscured under DMSO peak. LCMS m/z 392.08 [M+H]⁺.
Compound 120 _Oo>^ci Xx ^s n-nh ^h	S25; 0 H ^N'NH	Compound 21	‘H NMR (400 MHz, DMSOίΖ₆)δ 12.87 (s, 1H), 9.23 (d, J= 10.3 Hz, 1H), 8.72 (s, 1H), 6.94 (s, 1H), 6.09 (s, 1H), 4.44 (t, J = 11.1 Hz, 1H), 4.08 -3.82 (m, 2H), 3.52 (s, 1H), 2.63-2.57 (m, 2H), 2.44 - 2.36 (m, 1H), 2.23 (d, J= 14.4 Hz, 1H), 2.08 (t, J= 13.6 Hz, 1H), 1.92 (dt, J = 10.1,5.2 Hz, 1H), 1.86-1.74 (m, 1H), 1.26 (d, J =6.5 Hz,

216

Product	Starting Material and Aldéhyde	Method	‘HNMR; LCMS m/z [M+Hf
			3H), 1.02- 0.88 (m, 2H), 0.77 0.58 (m, 2H). LCMS m/z 364.13 [M+H]⁺.
Compound 121 <OlV^ci ⁰ II M ^H	S25; 0 0 A $ ^HN II	Compound 20	‘H NMR (400 MHz, DMSOJ₆) δ 12.10 (s, 1H), 8.99 (d,7 = 10.4 Hz, 1H), 8.49 (d, J= 11.0 Hz, 1H), 7.67 (dd, J= 7.0, 2.0 Hz, 1H), 7.49 (s, 1H), 6.93 (s, 1H), 6.32 (t, 7=6.7 Hz, 1H), 4.61 (t, 7= 11.2 Hz, 1H), 3.95 (t, 7=5.6 Hz, 2H), 3.58 (s, 1H), 2.60 (t, 7= 5.5 Hz, 2H), 2.24 (ddt, 7= 43.3, 26.7, 14.5 Hz, 3H), 1.94- 1.82 (m, 1H), 1.28 (d, 7= 6.5 Hz, 3H). LCMS m/z 351.17 [M+H]⁺.
Compound 122 <o>^{ci υ}χΑ Λ^Ν. -N H A N'ⁿ	S25; 0 l\| /-~ΝΊ n=n	Compound 21	LCMS m/z 379.12 [M+H]⁺.
Compound 123 oOi^^{ci s} F. fji fVⁿ	S25; 0 H E fV^N	Compound 20	‘H NMR (300 MHz, DMSOd₆) δ 9.10 (s, 1H), 8.96 (d,7= 2.1 Hz, 1H), 8.77 (s, 1H), 8.32 (dd, 7= 8.1, 2.2 Hz, 1H), 8.06 (d, 7=8.2 Hz, 1H), 6.95 (s, 1H), 4.80 (t, 7= 10.9 Hz, 1H), 3.98 (t, 7= 5.4 Hz, 2H), 3.69 (s, 1H), 2.62 (q, 7= 4.8 Hz, 2H), 2.34 (dd, 7=27.2, 14.5 Hz, 3H), 1.91 (t, 7= 13.3 Hz, 1H), 1.29 (d, 7=6.4 Hz, 3H). LCMS m/z 403.1 [M+H]⁺.
Compound 124 _oOlV^{ci ü}>c ^s ⁰ II A J ^H crV 1	S25; 0 0 ïY CTY 1	Compound 20	‘H NMR (400 MHz, DMSO7₆)δ8.91 (d, 7= 10.9 Hz, 1H), 8.48 (d, 7= 11.6 Hz, 1H), 8.03 (s, 1H), 6.94 (s, 1H), 4.43 (t,7 = 11.1 Hz, 1H), 3.91 (hept,7= 5.8,5.4 Hz, 2H), 3.53 (s, 1H), 3.36 (s, 3H), 3.22 (s, 3H), 2.60 (t, 7=5.5 Hz, 2H), 2.40-2.12 (m, 3H), 1.87- 1.75 (m, 1H), 1.26 (d, 7= 6.5 Hz, 3 H). LCMS m/z 396.15 [M+H]⁺.

217

Product	Starting Material and Aldéhyde	Method	'H NMR; LCMS m/z [M+Hf
Compound 125 ₀O>^ciyxff ^s L 1] H HO N	S25; HO^hT	Compound 21	‘H NMR (400 MHz, DMSOJ₆) δ 11.77 (s, 1H), 9.01 (s, 1H), 8.48 (s, 1H), 7.60 (s, 1H), 6.93 (s, 1H), 6.23 (s, 1H), 4.38 (t, 7= 11.4 Hz, 1H), 3.96 (t,7 = 5.2 Hz, 2H), 3.64 (s, 1H), 2.63 - 2.57 (m, 2H), 2.27 (dd, 7= 31.8, 14.9 Hz, 2H), 2.19 (s, 3H), 2.07 (t, 7= 13.5 Hz, 1H), 1.84 (t, 7= 13.3 Hz, 1H), 1.26 (d, 7= 6.4 Hz, 3H). LCMS m/z 365.13 [M+H]⁺.
Compound 126 oCO-d VpP Ί h	S24; 0 \.nJ TJ	Compound 20	’H NMR (400 MHz, DMSOd₆) δ 9.36- 9.17 (m, 1H), 8.80 (s, 1H), 7.79 (t, 7= 7.7 Hz, 1H), 7.35 (dd, 7= 19.9, 7.7 Hz, 2H), 6.94 (s, 1H), 4.62 (d,7 = 12.1 Hz, 1H), 4.07 - 3.92 (m, 2H), 3.57 (s, 1H), 2.62 (t, 7= 5.3 Hz, 2H), 2.56 (s, 3H), 2.35 (dd, 7=79.8, 14.4 Hz, 2H), 1.98 (dt, 7=35.2, 13.8 Hz, 2H), 1.32 (d, 7= 6.5 Hz, 3H). LCMS m/z 349.19 [M+H]⁺.
Compound 127 ₀O>-^cipyp	S25; ¹ ? ^x VŸ X	Compound 21	Ή NMR (400 MHz, DMSOd₆) δ 9.45 (s, 1H), 8.69 (s, 1H), 8.38 (d, 7= 5.1 Hz, 1H), 6.95 (s, 1H), 6.73 (d, 7= 5.0 Hz, 1H), 4.50 (t, 7= 11.4 Hz, 1H), 4.01 (dp, 7= 18.3, 6.2, 5.4 Hz, 2H), 3.63 - 3.53 (m, 1H), 3.18 (s, 6H), 2.63 (t, 7=5.4 Hz, 2H), 2.55 - 2.48 (m, hidden under DMSO, 1H), 2.26 (d, 7= 14.5 Hz, 1H), 1.95 (dt,7 = 25.4, 13.5 Hz, 2H), 1.34 (d,7 = 6.4 Hz, 3H). LCMS m/z 379.12 [M+H]⁺.
Compound 128 oOlV^ci HN^ ΰ N ^H	S25; 0 H H N 7^	Compound 21¹	Ή NMR (300 MHz, Methanol7₄) δ 7.62 (s, 1H), 6.76 (s, 1H), 4.91 (dd,7= 12.8,3.3 Hz, 1H), 4.01 (t, 7= 5.5 Hz, 2H), 3.83 (dddt,7= 13.2, 9.6,6.7,3.0 Hz, 1H), 3.00 (q, 7= 7.7 Hz, 2H), 2.67 (t, 7=5.6 Hz, 2H), 2.56 (dt, 7=14.4, 2.8 Hz, 1H), 2.46-2.33 (m, 2H), 1.90 (dd,7 = 14.8, 12.3 Hz, 1H), 1.45-

218

Product	Starting Material and Aldéhyde	Method	’HNMR; LCMS m/z [M+Hf
			1.35 (m, 6H). LCMS m/z 352.17 [M+H]⁺.
Compound 129 <OO^^ci /x A J ^H p N N _zfA	S25; 0 J fj p^N N /ⁿ A	Compound 21¹⁵	Ή NMR (400 MHz, DMSOde) δ 10.20 (s, 1H), 9.15 (s, 1H), 8.86 (s, 1H), 8.61 (s, 2H), 6.94 (s, 1H), 4.72 (s, 2H), 4.53 (t, 7= 11.5 Hz, 1H), 3.95 (t, J= 5.1 Hz, 2H), 3.61 (s, 1H), 3.56 -3.18(m, 4H), 3.12-3.02 (m, 2H), 2.84 (s, 3H), 2.61 (t, J= 5.0 Hz, 2H), 2.42 (t, J= 13.7 Hz, 1H), 2.32 - 2.22 (m, 2H), 1.87 (t, 7= 13.4 Hz, 1H), 1.27 (d, J= 6.5 Hz, 3H). LCMS m/z 434.16 [M+H]⁺.
Compound 130 oOl^^ci \=n ⁿ	S25; 0 H HNJ	Compound 21¹	Ή NMR (400 MHz, DMSOde) δ 7.36 (s, 1H), 6.94 (s, 1H), 4.50 (d, J= 11.1 Hz, 1H), 3.93 (t, J =5.5 Hz, 2H), 3.54 (s, 1H), 2.60 (s, 2H), 2.39 (d,7= 14.9 Hz, 1H), 2.23 (d, J= 13.9 Hz, 2H), 2.10 (s, 1 H), 1.871.75 (m, 1 H), 1.26 (d, J =6.4 Hz, 3H), 1.07 (s, 2H), 0.96 (s, 2H). Exchangeables not observed. LCMS m/z 364.13 [M+H]⁺.
Compound 131 (Oû^^cijxjs P \ M ^H / n-n	S25; 0 O'^N	Compound 21	’H NMR (400 MHz, DMSO7₆) δ 9.27 (d, 7=10.4 Hz, 1H), 8.76 (d, 7=10.9 Hz, 1H), 6.94 (s, 1H), 5.70 (s, 1H), 4.39 (t, 7 = 11.4 Hz, 1H), 4.29 (t, 7= 5.3 Hz, 2H), 4.09 (t, 7= 6.2 Hz, 2H), 3.94 (t, 7= 5.6 Hz, 2H), 3.57 - 3.47 (m, 1H), 2.60 (t, 7= 5.5 Hz, 2H), 2.39 (d, 7= 15.1 Hz, 1H), 2.23 (d, 7= 16.9 Hz, 1H), 2.17 (q, 7= 5.7 Hz, 2H), 2.11-2.01 (m, 1H), 1.80 (t, 7= 13.4 Hz, 1H), 1.26 (d,7=6.5 Hz, 3H). LCMS m/z 380.12 [M+H]⁺.

219

Product	Starting Material and Aldéhyde	Method	‘H NMR; LCMS m/z [M+II]
Compound 132 oOlV^ciz\ ^s NJ) H	S25; , o \ H wjf (A	Compound 21	Ή NMR (400 MHz, DMSOdd) δ 9.08 (d, 7=9.6 Hz, 1H), 9.01 (s, 1H), 8.77 (s, 1H), 6.94 (s, 1H), 4.36 (t, 7= 11.4 Hz, 1H), 3.95 (t, 7= 5.3 Hz, 2H), 3.66 (s, 1H), 2.66-2.58 (m, 2H), 2.40 (d, 7= 14.4 Hz, 1H), 2.32 (s, 3H), 2.22 (q,7= 14.1, 13.2 Hz, 2H), 1.82 (t, 7= 13.4 Hz, 1H), 1.27 (d, 7=6.5 Hz, 3H). LCMS m/z 339.16 [M+H]⁺.
Compound 133 (Oo^^c\| Nf J H °y_> h₂n	S25; 0 N J¹ N? J 0^_J h₂n	Compound 21	Ή NMR (400 MHz, DMSOdd) δ 9.39 (s, 1H), 8.93 (d,7= 10.9 Hz, 1H), 8.24 (s, 1H), 7.80 (s, 1H), 7.45 (s, 1H), 6.95 (s, 1H), 5.14 (s, 2H), 4.74 (t,7= 11.0 Hz, 1H), 3.96 (t,7=5.4 Hz, 2H), 3.61 (s, 1H), 2.62 (d, 7= 5.5 Hz, 2H), 2.47-2.17 (m, 3H), 1.89- 1.78 (m, 1H), 1.27 (d, 7= 6.5 Hz, 3H). LCMS m/z 382.15 [M+H]⁺.
Compound 134 oOlV^cijXJ s vL' ^H	S24; 0 /⁰A -A ii Vn	Compound 20	Ή NMR (400 MHz, DMSOdd) δ 9.70 (s, 1H), 9.31-9.09 (m, 1H), 7.00 (d, 7= 1.5 Hz, 1H), 6.95 (s, 1H), 4.75 (d,7= 11.5 Hz, 1H), 3.95 (t, 7= 5.4 Hz, 2H), 3.61 (s, 1H), 2.61 (t, 7 = 5.4 Hz, 2H), 2.33 (d,7= 1.3 Hz, 3H), 2.60 - 2.08 (m, 3H), 1.82 (dd, 7= 14.4, 12.2 Hz, 1H), 1.28 (d, 7=6.4 Hz, 3H). LCMS m/z 339.16 [M+H]⁺.
Compound 135 cOo^^c\| >1 N h i ^H	S25; 0 /O\/N3 TJ	Compound 20⁷	Ή NMR (400 MHz, DMSOdd) δ 9.40 (s, 1H), 8.68 (s, 1H), 7.81 (t, 7= 7.8 Hz, 1H), 7.17 (d, 7=8.0 Hz, 1H), 6.95 (s, 1H), 6.87 (d, 7=8.1 Hz, 1H), 4.61 (t, 7= 11.3 Hz, 1H), 3.95 (s, 5H), 3.66 - 3.55 (m, 1H), 2.63 (t, 7= 5.5 Hz, 2H), 2.492.44 (m, partly obstructed by DMSO, 1H), 2.28 (d,7= 14.4 Hz, 1H), 2.11 (t, 7= 13.4 Hz, 1H), 1.95 (t, 7= 13.3 Hz, 1H), 1.34 (d, 7= 6.4 Hz, 3H). LCMS m/z 365.17 [M+H]⁺.

220

Product	Starting Material and Aldéhyde	Method	‘HNMR; LCMS m/z [M+H]'
Compound 136 <On^^cizO⁵	S25; \ o \ l\| -~NJ hr	Compound 19	Ή NMR (400 MHz, DMSOd₆) δ 8.81 (s, 1H), 8.39 (d, J= 11.5 Hz, 1H), 7.54 (s, 1H), 6.93 (s, 1H), 4.34 (t, J= 11.2 Hz, 1H), 3.94 (t, 7=5.4 Hz, 2H), 3.73 (s, 3H), 3.64 (s, 1H), 2.59 (t, J= 5.4 Hz, 2H), 2.26 (s, 3H), 2.21 (dd, 7=23.1, 12.6 Hz, 3H), 1.88 - 1.77 (m, 1H), 1.24 (d, 7=6.6 Hz, 3H). LCMS m/z 352.21 [M+H]⁺.
Compound 137 (OlVci yxr' S \|l Ί ^H ' OH	S25; 0 OH	Compound 21	‘H NMR (400 MHz, DMSOJô) δ 9.77 (s, 1H), 9.08 (d,7= 10.4 Hz, 1H), 8.47 (s, 1H), 6.94 (s, 1 H), 6.61 (s, 1H), 6.52 (s, 1H), 6.3 8 (s, 1H), 4.38 (t, 7 = 11.4 Hz, 1H), 3.95 (t, 7= 5.4 Hz, 2H), 3.73 (s, 3H), 3.60 (s, 1H), 2.60 (t, 7= 5.4 Hz, 2H), 2.26 (d, 7= 13.9 Hz, 2H), 2.16 (t, 7= 13.4 Hz, 1H), 1.87 (t, 7= 13.3 Hz, 1H), 1.27 (d,7=6.4 Hz, 3H). LCMS m/z 380.16 [M+H]⁺.
Compound 138 oOCJ’⁰¹ l II N A IJ «	S25; ¹ H ΑΆ J H Ύ ^νΆ	Compound 21	‘H NMR (400 MHz, DMSOd₆) δ 9.38 (d, 7 = 10.2 Hz, 1H), 8.63 (s, 1H), 6.95 (s, 1H), 6.64 (s, 1H), 4.43 (t, 7 = 11.2 Hz, 1H), 4.09-3.91 (m, 2H), 3.683.50 (m, 1H), 3.17 (s, 6H), 2.63 (t, 7=5.4 Hz, 2H), 2.28 (s, 3H), 2.24 (d, 7= 12.6 Hz, 1H), 1.93 (dt, 7=21.5, 13.5 Hz, 2H), 1.33 (d, 7= 6.4 Hz, 3H), 1.16 (q, 7= 7.0 Hz, 1H). LCMS m/z 393.16 [M+H]⁺.
Compound 139 zz^sç/A	S25; 0 rj	Compound 19	LCMS m/z 349.23 [M+H]⁺.

221

Product	Starting Material and Aldéhyde	Method	LCMS m/z [M+Hf
Compound 140 cOlV^cijcjs N * H V-0 o—'	S25; 0 IJ Vo cJ	Compound 21¹⁴	’H NMR (400 MHz, DMSO7₆) δ 9.37 (s, 1H), 8.99 (s, 1H), 7.27 (s, 1H), 6.95 (s, 1H), 4.73 (t, 7= 11.0 Hz, 1H), 3.93 (t,7 = 5.5 Hz, 2H), 3.91 -3.84(m, 2H), 3.63 (s, 1 H), 3.46 (td,7= 11.5, 2.4 Hz, 2H), 3.16-3.04 (m, 1 H), 2.60 (dd, 7=8.4, 4.5 Hz, 2H), 2.47 - 2.42 (m, under DMSO, 1H), 2.23 (dt,7= 13.5, 6.2 Hz, 2H), 1.91 (d,7= 13.3 Hz, 2H), 1.83 (t, 7= 13.3 Hz, 1H), 1.78 - 1.65 (m, 2H), 1.26 (d, J= 6.5 Hz, 3H). LCMS m/z 409.11 [M+H]⁺.
Compound 141 oOCVa J ^hH 0	S25; 0 H Of o ^hH 0	Compound 21	*H NMR (400 MHz, DMSOd₆) δ 11.93 (s, 1H), 9.31 (s, 1H), 8.85 (s, 1H), 7.34 (d,7= 8.1 Hz, 1H), 7.26 (t, 7= 7.9 Hz, 1H), 7.20-7.12 (m, 1H), 6.94 (s, 1H), 4.78 (t, 7= 10.8 Hz, 1H), 3.99 (t, 7= 5.5 Hz, 2H), 3.71 (s, 1H), 2.62 (t, 7= 5.2 Hz, 2H), 2.46 - 2.21 (m, 3H), 1.92 (t, 7= 13.3 Hz, 1H), 1.29 (d, 7= 6.5 Hz, 3H). LCMS m/z 391.09 [M+H]⁺.
Compound 142 oCcvc JJ s NJ H hr /	S25; , o \ H KJ iv /	Compound 19	Ή NMR (400 MHz, DMSOdd) δ 8.80 (s, 1H), 8.38 (d, 7= 10.8 Hz, 1H), 7.79 (s, 1H), 6.94 (s, 1H), 4.31 (t, 7= 11.1 Hz, 1H), 3.94 (t, 7= 5.4 Hz, 2H), 3.76 (s, 3H), 3.62 (s, 1H), 2.60 (t, 7 = 5.5 Hz, 2H), 2.25 (d, 7 = 14.3 Hz, 2H), 2.19-2.10 (m, 4H), 1.83 - 1.74 (m, 1H), 1.25 (d, 7 = 6.7 Hz, 3H). LCMS m/z 352.21 [M+H]⁺.
Compound 143 oOlV^c\|n ci JA	S24; Γ15 ^NJ	Compound 20¹’⁶	*H NMR (400 MHz, DMSOd₆) δ 8.42 (dd, 7=6.8, 1.2 Hz, 1H), 7.92 (d, 7=1.3 Hz, 1H), 7.53 (d, 7=1.2 Hz, 1H), 7.25 (d, 7=6.9 Hz, 1H), 6.89-6.81 (m, 2H), 4.61 (dd,7 = 11.5, 2.5 Hz, 1H), 3.99 (hept, 7= 6.0 Hz, 2H), 3.18 (d, 7= 12.0 Hz, 1H), 2.59 (t, 7= 5.5 Hz, 2H), 2.51 1.90 (m, 2H), 1.41 (ddd,7 =

222

Product	Starting Material and Aldéhyde	Method	^]HNMR; LCMS m/z [M+Hf
			79.2, 13.2, 11.3 Hz, 2H), 1.07 (d, J= 6.3 Hz, 3H). LCMS m/z 374.16 [M+H]⁺.
Compound 144 <OlV^c' ’AA'S γϋ ^H	S25; 0 ~~^N	Compound 21	Ή NMR (400 MHz, DMSOJô) δ 9.70 (s, 1H), 9.17 (s, 1H), 6.95 (s, 1H), 4.70 (s, 1H), 3.94 (t, J =5.5 Hz, 2H), 3.68 - 3.53 (m, 1H), 2.60 (t, 7=5.4 Hz, 2H), 2.31 -2.08 (m, 6H), 2.082.04 (m, 3H), 1.86- 1.75 (m, 1H), 1.27 (d, 7=6.6 Hz, 3H). LCMS m/z 353.16 [M+H]⁺.
Compound 145 _oOüV^c\|ζχγ ^s~ ί χ Vn ^H	S25; 0 γλ/N^J XX	Compound 21²	Ή NMR (400 MHz, DMSO7₆) δ 9.59 (s, 1H), 8.99 (s, 1H), 8.63 (d, 7= 6.3 Hz, 1H), 7.00 (s, 1H), 6.95 (s, 1H), 4.60 (t, 7 = 11.0 Hz, 1H), 4.03 -3.93 (m, 2H), 3.97 (s, 3H), 3.58 (s, 1H), 2.74-2.56 (m, 3H),2.31 (d,7 = 14.7 Hz, 1H), 2.04- 1.93 (m, 1H), 1.93 - 1.81 (m, 1H), 1.35 (d, 7= 6.5 Hz, 3H). LCMS m/z 366.12 [M+H]⁺.
Compound 146 <QA^ciαΛ ri Y Ύ N ^° H 1 h	S25; 0 o 'W XX	Compound 21	Ή NMR (400 MHz, DMSOd₆) δ 9.16 (s, 1H), 8.73 (s, 1H), 8.16 (s, 1H), 8.00 (d, 7=7.8 Hz, 1H), 7.93 (d, J= 7.8 Hz, 1H), 7.75 (t, 7= 7.8 Hz, 1H), 6.95 (s, 1H), 4.73 (t, 7= 11.2 Hz, 1H), 3.98 (t, 7= 5.4 Hz, 2H), 3.68 (s, 1H), 3.25 (s, 3H), 2.61 (d, 7=4.5 Hz, 2H), 2.39 (d, J= 14.5 Hz, 1H), 2.30 (t, J = 13.5 Hz, 2H), 1.91 (t,7 = 13.3 Hz, 1H), 1.29 (d,7=6.5 Hz, 3H). LCMS m/z 412.09 [M+H]⁺.
Compound 147 oQl>⁰¹ h₂n jfj H	S25; h₂n Έ J	Compound 20	Ή NMR (300 MHz, DMSOd₆) δ 9.12 (d, J= 10.8 Hz, 1H), 8.72 - 8.54 (m, 1H), 8.05 (d, 7 = 1.8 Hz, 1H), 7.92-7.87 (m, 1H), 7.79 (d, 7= 7.8 Hz, 1H), 7.67 (t, 7= 7.7 Hz, 1H), 7.44 (s, 2H), 6.94 (s, 1H), 4.67 (t, 7 = 10.5 Hz, 1H), 3.97 (t,7=5.4 Hz, 2H), 3.68 (s, 1H), 2.65 2.58 (m, 2H), 2.35 - 2.24 (m, 3H), 1.90 (t, 7= 13.3 Hz, 1H),

223

Product	Starting Material and Aldéhyde	Method	flNMR; LCMS m/z[M+Hf
			1.29 (d, 7= 6.4 Hz, 3H). LCMS m/z 413.12 [M+H]⁺.
Compound 148 <On^^{ci h} ζ 7 N-NH	S25; 0 H ζ / N-NH	Compound 21	’H NMR (400 MHz, DMSOd₆) δ 9.16 (s, 1H), 8.73 (s, 1H), 8.11 (s, 2H), 7.82 (s, 1H), 7.67 (d, 7= 7.5 Hz, 1H), 7.44 (t, 7= 7.6 Hz, 1H), 7.38 (d,7=7.7 Hz, 1H), 6.94 (s, 1H), 4.61 4.47 (m, 1H), 3.98 (t, 7= 5.5 Hz, 2H), 3.69 - 3.59 (m, 2H), 2.62 (s, 2H), 2.39 - 2.24 (m, 3H), 1.94 (t, 7= 13.3 Hz, 1H), 1.30 (d, 7= 6.4 Hz, 3H). LCMS m/z 400.12 [M+H]⁺.
Compound 149 <OO^^ci Xx ^s O⁷	S25; 0 H X/ ^-N (J	Compound 21	‘H NMR (300 MHz, MethanolJ₄) δ 7.77 - 7.54 (m, 1H), 6.77 (s, 1H), 4.96 (s, 3H), 4.18 (s, 4H), 4.03 (t, 7= 5.4 Hz, 2H), 3.98 -3.84 (m, 1H), 2.67 (t,7= 5.5 Hz, 2H), 2.58 (d,7= 14.8 Hz, 1H), 2.42 (d, 7= 15.9 Hz, 1H), 2.37-2.23 (m, 1H), 1.88 (s, 1H), 1.41 (d, 7= 6.5 Hz, 3H). LCMS m/z 380.12 [M+H]⁺.
Compound 150 oOlV^ci H fl N A_na, ^H	S25; H 5 N^é Vn	Compound 21¹	Ή NMR (400 MHz, DMSO7₆) δ 6.95 (s, 1H), 4.58 (s, 1H), 3.95 (t, 7= 5.5 Hz, 2H), 3.583.45 (m, 1H), 2.64 - 2.55 (m, 4H), 2.22 (d, 7= 14.4 Hz, 1H), 2.13 (s, 6H), 1.77 (t, 7= 13.3 Hz, 1H), 1.26 (d, 7= 6.5 Hz, 3H). Exchangeables not observed. LCMS m/z 352.17 [M+H]⁺.
Compound 151 ₀O>^ci >çs H \|\| ^Ο^Ζ^Ν^μΑ é N L IJ H	S24; H U °V^N\Z XJ	Compound 20	’H NMR (300 MHz, Methanol7₄) δ 7.69 (t, 7= 7.9 Hz, 1H), 6.86 (d, 7=7.3 Hz, 1H), 6.75 (s, 1H), 6.68 (d, 7=8.4 Hz, 1H), 4.64 (dd, 7= 12.4, 3.0 Hz, 1H), 4.03 (t, 7= 5.5 Hz, 2H), 3.83 -3.72 (m, 1H), 2.67 (t, 7= 5.5 Hz, 2H), 2.57 - 2.45 (m, 1H), 2.38 (dt, 7= 14.5, 2.6 Hz, 1H), 2.14 (dd, 7= 14.1, 12.9 Hz, 1H), 1.86 (dd, 1H), 1.41 (d, J= 6.6 Hz, 3H). LCMS m/z 351.17 [M+H]⁺.

224

Product	Starting Material and Aldéhyde	Method	^!HNMR; LCMS m/z [M+H]'
Compound 152 éjpé éé~s \ il O »	S25; ' ? N-é ⁿu	Compound 19¹⁷	NMR not integrated due to a complex mixture of stereoisomers LCMS m/z 338.21 [M+H]⁺.
Compound 153 ₀O>^ciéé / V-NH ^H	S25; 0 / Xt-NH	Compound 21¹	*H NMR (400 MHz, DMSOJ₆) δ 6.99 (s, 1H), 6.95 (s, 1H), 4.60 (d, J= 12.2 Hz, 1H), 3.96 (t, 7= 5.5 Hz, 2H), 3.6(1 H under water peak), 2.65 - 2.47 (m, 5H), 2.23 (d, 7=14.4 Hz, 1H), 2.11 (t, 7= 13.3 Hz, 1H), 1.82 (t, 7= 13.2 Hz, 1H), 1.28 (d, 7=6.5 Hz, 3H), 1.18 (t, 7 = 7.5 Hz, 3H). LCMS m/z 352.17 [M+H]⁺.
Compound 154 oOTé ^U><L ^sil JJé é /	S25; Ά ÿ kj é /	Compound 21	Ή NMR (400 MHz, DMSOde) δ 8.94 (s, 1H), 8.51 (s, 1H), 7.70 (s, 1H), 6.93 (s, 1H), 4.32 - 4.22 (m, 1H), 3.92 (t, 7= 5.5 Hz, 2H), 3.83 (s, 3H), 3.70 (s, 3H), 3.59 - 3.48 (m, 1H), 2.59 (t, 7= 5.4 Hz, 2H), 2.27-2.18 (m, 3H), 1.82-1.71 (m, 1H), 1.23 (d, 7= 6.4 Hz, 3H). LCMS m/z 368.15 [M+H]⁺.
Compound 155 oOlVci nY_nX H	S24; 0 V Vo	Compound 20	Ή NMR (400 MHz, DMSO7₆) δ 9.92 - 9.60 (m, 1H), 9.35 9.09 (m, 1H), 8.01 - 7.95 (m, 1H), 6.95 (s, 1H), 4.78 (s, 1H), 3.95 (t, 7= 5.4 Hz, 2H), 3.61 (s, 1 H), 2.60 (q, 7=5.1 Hz, 3H), 2.25 (d, 7 =14.3 Hz, 1H), 2.13 (d, 7= 1.3 Hz, 3H), 2.18 2.06 (m, 1H), 1.87- 1.75 (m, 1H), 1.28 (d, 7=6.4 Hz, 3H). LCMS m/z 339.16 [M+H]⁺.
Compound 156 <OTé jxjs ,νΤ,Α __// é N \ II H Hré	S24; 0 é Hé	Compound 20	’H NMR (400 MHz, DMSOde) δ 9.17 (s, 1H), 9.01-8.60 (m, 1H), 8.32 (s, 1H), 6.94 (s, 1H), 4.56 (d, 7= 12.2 Hz, 1H), 3.94 (t, 7= 5.4 Hz, 2H), 3.693.61 (m, 1H), 2.60 (t, 7= 5.4 Hz, 2H), 2.45 - 2.33 (m, 1H), 2.27 (s, 3H), 2.25 (s, 2H), 1.83 (dd, 7=14.4, 12.3 Hz, 1H),

225

Product	Starting Material and Aldéhyde	Method	’HNMR; LCMS m/z [M+Ilf
			1.25 (d, 7= 6.5 Hz, 3H). LCMS m/z 338.17 [M+H]⁺.
Compound 157 oOA^CI AA ÂAa Vo ^M	S25; \ ⁰A t-o	Compound 21	‘H NMR (400 MHz, DMSO7₆) δ 9.22 (s, 1H), 8.92 (s, 1H), 8.44 (s, 1H), 6.94 (s, 1H), 4.75 (s, 1H), 3.94 (t, 7=5.6 Hz, 2H), 3.70 (s, 1H), 2.60 (t, 7 = 5.5 Hz, 2H), 2.36 - 2.22 (m, 3H), 2.17 (s, 3H), 1.84 (t, 7 = 13.4 Hz, 1H), 1.24 (d,7=6.6 Hz, 3H). LCMS m/z 339.12 [M+H]⁺.
Compound 158 (OCVci AA'³ n'a N HO	S24; < 0 \ H ⁿAT hr HO	Compound 20	Ή NMR (300 MHz, Methanol70 δ 7.80 (s, 1H), 6.75 (s, 1H), 4.56 (dd,7= 12.6, 2.8 Hz, 1H), 4.16 (t, 7= 5.2 Hz, 2H), 4.01 (t, 7=5.5 Hz, 2H), 3.84 (q,7= 5.5 Hz, 3H), 2.66 (t, 7= 5.4 Hz, 2H), 2.45 - 2.34 (m, 2H), 2.29 (s, 3H), 2.15 (dd, 7= 14.8, 12.6 Hz, 1H), 1.80 (dd, 7 = 14.9, 12.2 Hz, 1H), 1.37 (d, 7=6.6 Hz, 3H). LCMS m/z 382.15 [M+H]⁺.
Compound 159 (O0^^ci AA^S Rr Γ \=N	S24; 0 Π —N7	Compound 20¹²	Ή NMR (300 MHz, Methanol70 δ 7.79 (s, 1H), 6.76 (s, 1H), 4.94-4.89 (dd, under water, 1H), 4.06-3.90 (m, 3H), 3.83 (s, 3H), 2.70 - 2.64 (m, 5H), 2.55 (dt, 7= 14.6, 2.7 Hz, 1H), 2.46 -2.28 (m, 2H), 1.92 (dd,7 = 14.8, 12.3 Hz, 1H), 1.42 (d,7 = 6.6 Hz, 3H). LCMS m/z 352.17 [M+H]⁺.
Compound 160 R ^U>A oh il Ô - N	S25; OH 0 σ	Compound 21	‘H NMR (400 MHz, DMSO70 δ 11.99 (s, 1H), 9.00 (s, 1H), 8.52 (s, 1H), 7.95 (s, 1H), 7.78 (d, 7=7.4 Hz, 1H), 6.93 (s, 1H), 6.27 (d, 7=7.3 Hz, 1H), 4.64-4.54 (m, 1H), 3.94 (t, 7= 5.5 Hz, 2H), 3.6(1 H under water peak), 2.60 (t, 2H), 2.23 (q, 7= 12.2, 11.0 Hz, 3H), 1.93 - 1.83 (m, 1H), 1.28 (d,7 = 6.4 Hz, 3H). LCMS m/z 351.13 [M+H]⁺.

226

Product	Starting Material and Aldéhyde	Method	'H NMR; LCMS m/z [M+HJ
Compound 161 <OlVci AA H il NxAmA / Pf N \\ II H N^\	S24; H 5 z^NA < T ^Ν\	Compound 20	Ή NMR (400 MHz, DMSOd₆) δ 9.12 (s, 1H), 8.95-8.65 (m, 1H), 8.30 (s, 1H), 6.94 (s, 1H), 4.56 (d, J= 12.2 Hz, 1H), 3.94 (t, 7=5.4 Hz, 2H), 3.65 (s, 1H), 2.63 - 2.57 (m, 2H), 2.44 - 2.34 (m, 1H), 2.27 (s, 3H), 2.23 (d, 7= 13.0 Hz, 2H), 1.90- 1.76 (m, 1H), 1.25 (d,7 = 6.5 Hz, 3H). LCMS m/z 340.2 [M+H]⁺.
Compound 162 oAlA^ci\/N A_NA ^H	S25; 0 \/ⁿA / Xï-NH	Compound 21¹	Ή NMR (300 MHz, Methanol7₄) δ 7.19 (s, 1H), 6.76 (s, 1H), 4.97 - 4.88 (m, under water, 1H), 4.01 (t, 7=5.5 Hz, 2H), 3.76 (s, 1H), 3.03 (dd,7=13.4, 7.0 Hz, 1H), 2.66 (t, J= 5.5 Hz, 2H), 2.57 (s, 1H), 2.35 (d,7= 14.8 Hz, 2H), 1.87 (t, 7= 13.3 Hz, 1H), 1.39 (d, 7= 6.8 Hz, 3H), 1.30 (dd, 7= 7.0, 1.6 Hz, 6H). LCMS m/z 366.16 [M+H]⁺.
Compound 163 Γτα Ax ^S Ά B nA AJ³	S25; 0 H NJ nA AJ³	Compound 21	‘H NMR (400 MHz, DMSO7₆) δ 8.93 (s, 1H), 8.50 (s, 1H), 7.45 (s, 1H), 6.93 (s, 1H), 4.34 (s, 2H), 4.26 (t, 7= 11.4 Hz, 1H), 4.07 (t, 7= 6.0 Hz, 2H), 3.91 (t, 7= 5.4 Hz, 2H), 3.51 (s, 1H), 2.59 (t, 7= 5.4 Hz, 2H), 2.22 (dd, 7=29.3, 12.9 Hz, 5H), 1.84- 1.71 (m, 1H), 1.23 (d, 7= 6.7 Hz, 3H). LCMS m/z 380.12 [M+H]⁺.
Compound 164 oOO^^ciAx ^s N ιΐ^Ν^' M ^H	S25; 1 o 1 H γ iT	Compound 21	Ή NMR (400 MHz, DMSOde) δ 9.40 (s, 1H), 9.00 (d,7= 12.1 Hz, 1H), 8.61 (q,7=2.7 Hz, 2H), 6.95 (s, 1H), 4.94 (t, 7 = 11.2 Hz, 1H), 4.04 (t, 7= 5.4 Hz, 2H), 3.78-3.61 (m, 1H), 2.63 (d, 7=5.4 Hz, 2H), 2,6 (1H under DMSO peak) 2.58 (s, 3H), 2.24 (dd, 7= 14.0, 10.2 Hz, 1H), 2.08 - 1.79 (m, 2H), 1.32 (d, 7=6.6 Hz, 3H). LCMS m/z 350.14 [M+H]⁺.

227

Product	Starting Material and Aldéhyde	Method	‘HNMR; LCMS m/z [M+Hf
Compound 165 (OO^^ciζχχ-s F ^H J 1 f—t_/ Tr ^N Γ \\ ¹¹ H F V-N	S25; F H ? 1 ^F~H v Vn	Compound 21	*H NMR (400 MHz, DMSOd₆) δ 13.16 (s, 1H), 9.61 (s, 1H), 9.16 (s, 1H), 7.94 (s, 1H), 6.96 (s, 1H), 4.70 (t, J= 10.9 Hz, 1H), 3.97 (t, J =5.5 Hz, 2H), 3.56 (s, 1H), 2.62 (t, 7= 5.8 Hz, 2H), 2.56 (s, 1H), 2.24 (d, 7 = 14.4 Hz, 1H), 2.06 (t,7 = 13.4 Hz, 1H), 1.93 - 1.82 (m, 1H), 1.30 (d, 7= 6.4 Hz, 3H). LCMS m/z 392.08 [M+H]⁺.
Compound 166 oOCVc oh II Il Ί ^H	S25; OH 0 X	Compound 21	'H NMR (300 MHz, MethanolJ4) δ 7.12 (dd, 7= 8.2, 1.6 Hz, 1H), 7.01 (t, 7=7.9 Hz, 1H), 6.94 - 6.89 (m, 2H), 4.90 (t, 7= 2.9 Hz, 1H), 3.88 (s, 3H), 3.81 (td, 7=6.7, 1.5 Hz, 2H), 3.423.30 (m,lH), 3.03 -2.85 (m, 2H), 2.66 (td, 7= 14.5, 3.3 Hz, 2H), 2.44 (dt, 7 = 14.2, 2.6 Hz, 1H), 2.18 (dd, 7= 14.5, 12.3 Hz, 1H), 1.31 (d, 7= 6.5 Hz, 3H). LCMS m/z 380.16 [M+H]⁺.
Compound 167 (OlVci zA⁸ A·	S25; 1 0 1 H pïl A/N	Compound 20	Ή NMR (400 MHz, DMSOJ₆) δ 9.26 (d, 7= 9.9 Hz, 1H), 8.94 (d, 7= 11.5 Hz, 1H), 8.56 - 8.50 (m, 1H), 7.76 - 7.70 (m, 1H), 7.40 (dd, 7=7.7, 4.8 Hz, 1H), 6.94 (s, 1H), 4.84 (t, 7 = 11.3 Hz, 1H), 4.02 (t, 7= 5.4 Hz, 2H), 3.65 (dq, 7=20.5, 5.7, 5.0 Hz, 1H), 2.62 (dt, 7= 5.5,3.2 Hz, 2H), 2.34 (s, 3H), 2.47-2.19 (m, 2H), 1.93 (ddd, 7=66.7, 14.5, 12.1 Hz, 2H), 1.32 (d, 7= 6.5 Hz, 3H). LCMS m/z 349.19 [M+H]⁺.
Compound 168 oOlA^ciz\ ^s L II h Vnh ^=N	S25; O H Of N H ^=N	Compound 21²	NMR not annotated due a complex mixture of stereoisomers. LCMS m/z 374.11 [M+H]⁺.

228

Product	Starting Material and Aldéhyde	Method	'HNMR; LCMS m/z [M+Hf
Compound 169 _oOlV^ci ^hnCT h O	S24; 0 ,^NHN J O	Compound 20	‘H NMR (300 MHz, Methanol7₄) δ 7.79 (d, 7= 8.2 Hz, 1H), 7.55 (d, 7= 8.5 Hz, 1 H), 7.42 (dd, 7= 8.1, 6.9 Hz, 1H), 7.21 (t, 7=7.6 Hz, 1H), 6.76 (s, 1H), 5.21 (dd, 7= 12.5, 3.0 Hz, 1H), 4.10 (t, 7= 5.6 Hz, 2H), 3.89 (dtq, 7= 12.6,6.2,3.1 Hz, 1H), 2.73 - 2.61 (m, 3H), 2.50 2.37 (m, 2H), 2.34 (dd,7 = 14.6, 12.7 Hz, 1H), 1.93 (dd,7 = 14.8, 12.3 Hz, 1H), 1.42 (d,7 = 6.6 Hz, 3H). LCMS m/z 374.16 [M+H]⁺.
Compound 170 oOlVo ~Xp^s L II h \P^N°A	S25; 0 H q N °A	Compound 2O¹⁰	LCMS m/z 389.1 [M+H]⁺.
Compound 171 oOlV^cipXp'S / N \\ M ^H N^N\	S24; 0 N^J V^N _X	Compound 20¹²	‘H NMR (400 MHz, DMSOd₆) δ 9.52 (s, 1H), 9.17 (s, 1H), 8.09 (s, 1H), 6.95 (s, 1H), 4.98 (s, 1H), 3.99 (t, 7= 5.4 Hz, 2H), 3.90 (d, 7=5.2 Hz, 3H), 3.69 (d, 7= 10.8 Hz, 1H), 2.71 - 2.56 (m, 3H), 2.32 - 2.05 (m, 2H), 1.97- 1.86 (m, 1H), 1.28 (d, 7= 6.5 Hz, 3H).. LCMS m/z 339.16 [M+H]⁺.
Compound 172 <OlV^ci ,υΑΑ L II h ^ΌΗ	S25; O Γ X	Compound 20	‘H NMR (300 MHz, DMSOd₆) δ 10.67 (s, 1H), 9.17 (d, 7= 8.6 Hz, 1H), 8.91 (d,7= 12.0 Hz, 1H), 8.16 (dd, 7= 3.9, 2.1 Hz, 1H), 7.37 - 7.27 (m, 2H), 6.93 (s, 1H), 4.85 (d,7= 11.6 Hz, 1H), 3.99 (s, 2H), 3.59 (s, 1H), 2.63 (t, 7=5.5 Hz, 2H), 2.44 (s, 1H), 2.26 (d,7 = 14.1 Hz, 1H), 2.02-1.91 (m, 1H), 1.90- 1.80 (m, 1H), 1.32 (d, 7 = 6.4 Hz, 3H). LCMS m/z 351.17 [M+H]⁺.

1. The product was afforded as the free-base.

229

2. Product was isolated as a 3:2 mixture of diastereomers with unknown absolute stereochemistry.

3. The mixture from step 1 was blown down with nitrogen at 40 °C.

4. Purification by silica gel chromatography (Gradient: 0-100% of 20% MeOH/DCM in DCM) yielded the product.

5. The TB S was deprotected during the reaction (step 2).

6. The product was impure after the purification and was repurified by reversedphase HPLC (Method: Cl8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 10 mM ammonium hydroxide).

7. Product was isolated as a 4.5:1 mixture of diastereomers with unknown absolute stereochemistry.

8. Step 1 was stirred at room température for one week.

9. The pH was carefully adjusted to pH 7 with 2 N NaOH prior to DCM extraction.

10. Product was isolated as a 2:1 mixture of diastereomers with unknown absolute stereochemistry.

11. Product was isolated as a 3:1 mixture of diastereomers with unknown absolute stereochemistry.

12. Product was isolated as a 5:1 mixture of diastereomers with unknown absolute stereochemistry.

13. Product was isolated as a 3.5:1 mixture of diastereomers with unknown absolute stereochemistry.

14. The product was impure after the purification and was repurified by reversedphase HPLC (Method: Cl8 Waters Sunfire column (30 x 150 mm, 5 micron).

Gradient: MeCN in H2O with 0.2% formic acid). The product was afforded as the formic acid sait.

15. The product was afforded as the di-TFA sait.

16. Product was isolated as a 4:1 mixture of diastereomers with unknown absolute stereochemistry.

17. Complex mixture of stereoisomers was afforded.

Compound 173 (2S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'~ piperidine] (173)

230

Step 1. Synthesis of l-[(2S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[6,7dihydrothieno[3,2-c]pyran-4,4 ’-piperidine]-1 '-yl]-2,2,2-trifluoro-ethanone (C61)

To a solution of 2-(5-chloro-2-thienyl)ethanol S2 (410 mg, 2.521 mmol) and (2S)-2methyl-6-(l-methyltriazol-4-yl)piperidin-4-one C56 (420 mg, 2.141 mmol) in DCM (8 mL) was added methanesulfonic acid (800 pL, 12.33 mmol). The resulting mixture was heated to 40 °C for 40 minutes. More methanesulfonic acid (800 pL, 12.33 mmol) was added and the reaction was continued heating for another 30 minutes. The reaction was cooled to room température, diluted with water, and basified with 2 N NaOH solution. The mixture was extracted with DCM (3 x 20 mL) through a phase separator and the organic layers were concentrated in vacuo to afford crude (2S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[6,7-dihydrothieno[3,2c]pyran-4,4'-piperidine]

A solution of crude (2S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[6,7dihydrothieno[3,2-c]pyran-4,4'-piperidine] in DCM (9 mL) with DIPEA (600 pL, 3.445 mmol) was cooled to 0 °C. TF AA (390 pL, 2.806 mmol) was added slowly over 2 minutes, and the reaction was stirred at 0 °C. After 15 minutes, the reaction was quenched with saturated sodium bicarbonate solution and extracted with DCM (3x). The organics were dried over sodium sulfate and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-50% EtOAc in Heptane) afforded a single major product l-[(2S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4yl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C61 (450 mg, 43%). Ή NMR (300 MHz, Chloroform-tZ) δ 7.58 (s, 1H), 6.91 (s, 1H), 5.57 (s, 1H), 4.40 (d, J= 7.4 Hz, 1H), 4.10 (s, 3H), 3.89 (t, J= 5.4 Hz, 2H), 3.20 (dd, J= 14.9, 6.4 Hz, 1H), 2.80 2.61 (m, 2H), 2.45 (dd, J= 14.8, 8.4 Hz, 1H), 2.38 - 2.13 (m, 1H),2.O4 (s, 1H), 1.41 - 1.12 (m, 3H).

Step 2. Synthesis of (2S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[6,7dihydrothieno[3,2-c]pyran-4,4'-piperidine] (173)

To a solution of l-[(2S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[6,7dihydrothieno[3,2-c]pyran-4,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C61 (20 mg, 0.04415 mmol) in MeOH (1 mL) was treated with NaOH (400 pL of 2 M, 0.8000 mmol). The solution

231 was heated to 50 °C for 3 hours at which point it was cooled to room température and stirred ovemight. The reaction was extracted with DCM (3x) through a phase separator and the organics concentrated in vacuo to afford (2S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[6,7dihydrothieno[3,2-c]pyran-4,4'-piperidine] 173 (14.0 mg, 91%) as an off-white film with an approximate e.r. of 85%. *H NMR (300 MHz, Chloroform-J) δ 7.41 (s, 1H), 6.61 (s, 1H), 4.40 (dd, J= 11.8, 2.7 Hz, 1H), 4.05 (s, 3H), 3.96 (td, J= 5.7, 2.0 Hz, 2H), 3.28 (dtd, J= 12.6, 6.3, 2.5 Hz, 1H), 2.85 - 2.60 (m, 2H), 2.18 (dt, J= 13.5, 2.6 Hz, 1H), 1.89 (dt, J= 13.7, 2.5 Hz, 1H), 1.80 (dd, 7=13.6, 11.9 Hz, 1H), 1.44 (dd,7=13.7, 11.4 Hz, 1H), 1.11 (d,7=6.3 Hz, 3H). LCMS m/z 339.1 [M+H]⁺

Préparation of S32 (2S,4S,6S)-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2-trifluoroacetyl)-2'(trifluoromethyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one (S32)

S32

Step 1. Synthesis of2,2,2-trifluoro-l-[(2'S,6'S,7S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4’-piperidine]-1 ’-yl]ethanone (C62)

To a solution of (2'S,6'S,7S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] 2 (1260 mg, 3.352 mmol) dissolved in DCM (25 mL) cooled to -15 °C was added DIPEA (800 pL, 4.593 mmol) followed by TFAA (550 pL, 3.957 mmol). After 5 minutes, the mixture was quenched with 1 N HCl (25 mL) and the phases were separated. The organic layer was dried with MgSÜ4, filtered, and

232 concentrated. Purification by silica gel chromatography (Gradient: 0-50% EtOAc in Heptane) yielded 2,2,2-trifluoro-l-[(2'S,6'S,7S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]ethanone C62 (1444 mg, 90%). Ή NMR (400 MHz, Methanol-ck) δ 7.93 (s, 1H), 7.27 (d, 7= 1.3 Hz, 1H), 5.63 (s, 1H), 4.46 (h, 7= 7.1 Hz, 1H), 4.11 (d, 7= 1.4 Hz, 3H), 3.96 (td, 7= 5.6, 1.7 Hz, 2H), 3.04 (s, 1H), 2.79 - 2.70 (m, 3H), 2.51 (s, 1H), 2.09 (dd, 7= 14.7, 7.3 Hz, 1H), 1.23 (q, 7= 9.6, 8.4 Hz, 3H). LCMS m/z 469.14 [M+H]⁺.

Step 2. Synthesis of (2S,4S,6S)-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2-trifluoroacetyl)2 '-(trifluoromethyl)spiro[piperidine-4,7'-thieno[2,3-cJpyran]-4 '-one (S32)

To a mixture of 2,2,2-trifluoro-l-[(2'S,6'S,7S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]ethanone C62 (708 mg, 1.511 mmol) in acetonitrile (10 mL) was added N-hydroxyphthalimide (165 mg, 1.011 mmol) and cobaltous diacetate tetrahydrate (35 mg, 0.1405 mmol), and then the mixture was vacuum purged with an oxygen balloon three times. The mixture was heated to 60 °C and stirred. After an hour and a half the reaction was cooled to room température. The mixture was vacuum purged with nitrogen three times and then diluted with MTBE (25 mL) and saturated aqueous bicarbonate (25 mL). The layers were separated, and the organic layer was washed with aqueous NaHCCh (2 x 50 mL) and brine (50 mL). The organic layer was dried with NaaSCU, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-50% EtOAc in heptane) afforded (2S,4S,6S)-2-methyl-6-(l-methyltriazol-4-yl)-1-(2,2,2-trifluoroacetyl)-2'(trifluoromethyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S32 (207 mg, 26%), *H NMR (300 MHz, Methanol-^) δ 7.98 (s, 1H), 7.80 (d, 7= 1.4 Hz, 1H), 5.70 (s, 1H), 4.48 (s, 1H), 4.45 (s, 2H), 4.12 (s, 3H), 2.95 (dd, 7= 14.8, 9.8 Hz, 1H), 2.73 (s, 1H), 2.22 (dd, 7= 14.8, 8.4 Hz, 1H), 1.29 (s, 1H), 1.19 (d, 7= 14.9 Hz, 3H). LCMS m/z 483.45 [M+H]⁺.

Préparation of Intermediates S33-S36

Intermediate ketones S33-S36 (see Table 4) were prepared in two steps from the relevant compounds using a TF AA protection and benzylic oxidation as described for intermediate S32. Any modifications to methods are noted in Table 4 and accompanying footnotes.

233

Table 4. Method of préparation, structure and physicochemical data for ketone intermediates S33-S36

Product	Compound Starting Material	¹ II NMR; LCMS m/z [M+Hf
o Μ ω - u_ U. ( O Z	SM κΑ_μΛ, N 3 H / Compound 1²>³’⁴,9	Ή NMR (300 MHz, Chloroformd) δ 7.63 (s, 1H), 7.21 (s, 1H), 5.63 (s, 1H), 4.45 (q, J= 7.5 Hz, 1H), 4.33 (s, 2H), 4.14 (s, 3H), 3.36 (dd, J= 15.1, 6.3 Hz, 1H), 2.80 (dd, J= 15.5, 8.7 Hz, 1H), 2.05 (d, J= 11.0 Hz, 2H), 1.27 (s, 3H). LCMS m/z 449.04 [M+H]⁺.
°\ ?—\ m Vz V Vo g >^/=\ * “π π ^r ω x) o	o00^^ci MM s ΝχλΑ O H M / Compound 22^,5,2,6,7,9,10	LCMS m/z 448.0 [M+H]⁺.
S35 0 oJlV^ciSM⁵ Z/S M Ai	ΜίΜ^αMjS n.st h M / Compound 19¹,²,⁵,⁸,⁹’¹¹	LCMS m/z 448.05 [M+H]⁺
S36 0 sj\ ^Cl CM —Mi'' y n-n A,.f u	^ciMX 0 —fMM N ’ H M ^m Compound 1731,8,9,12,13,¹⁴	*H NMR (300 MHz, Chloroformé δ 5.80 - 5.44 (m, 1H), 7.47 - 6.97 (m, 1H), 7.53 (d, J = 42.1 Hz, 1H), 4.42 (q, J= 7.2 Hz, 1H), 4.33 (d, J = 2.6 Hz, 2H), 4.13 (s, 3H), 3.36 (dd, J= 14.9, 5.2 Hz, 1H), 2.64 (dd, J= 15.1, 8.6 Hz, 1H), 2.46 (s, 1H), 2.16 (s, 1H), 1.32 (d, J=7.0Hz, 3H). LCMS m/z 449.12 [M+H]⁺

1. TF AA was added at 0 °C (step 1)

234

2. Reaction was stirred at 45 °C (step 2)

3. The mixture was quenched with water (10 mL) and the layers were separated. The organic layer was washed with 1 N HCl (10 mL), brine (10 mL), dried with magnésium sulfate, filtered, and concentrated. (step 1)

4. Reaction stirred for 18 hours (step 2)

5. Purification by silica gel chromatography (0-100% EtOAc:heptane) yielded the product (step 1)

6. Quenched with water before 1 N HCl (step 1)

7. Reaction stirred for 45 minutes (step 1)

8. Reaction was diluted with DCM, water, and saturated sodium bicarbonate. Extracted with DCM (3x) and collected through a phase separator (step 1)

9. Reaction was diluted with DCM, water, and saturated sodium bicarbonate. Extracted with DCM (3x) and collected through a phase separator (step 2)

10. Purification by silica gel chromatography (0-100% EtOAc in heptane) yielded the product (step 2)

11. Purification by silica gel chromatography (0-80% EtOAc in heptane) yielded the product (step 2)

12. Purification by silica gel chromatography (0-45% EtOAc in heptane) yielded the product (step 2)

13. Reaction stirred ovemight (step 2)

14. S36 has an approximately 85% e.r.

235

Préparation of S33

S33

Step 1. Synthesis of l-[(2'S,6'S,7S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-1 '-yl]-2,2,2-trifluoro-ethanone (Cl54)

To a mixture of (2'S,6'S,7S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] 1 (15.0 g, 43.82 mmol) and DIPEA (10 mL, 57.41 mmol) in DCM (150 mL), cooled to 3°C, was added TF AA (6.4 mL, 46.04 mmol). After 5 minutes, the mixture was quenched with 1 N HCl (100 mL), and the phases were separated. The organic layer was washed with brine (100 mL), dried with magnésium sulfate, filtered, and concentrated. The solid was suspended in TBME (100 mL) and heated to reflux. After 30 minutes, the mixture was cooled to 0 °C, and after 10 minutes, the material was filtered and rinsed with additional cold TBME. The product was dried to yield l-[(2'S,6'S,7S)-2-chloro-2'methyl-6'-(l-methyltriazol-4- yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]2,2,2-trifluoro-ethanone C154 (15.532 g, 81%). LCMS m/z cale. 435.18 [M+H]⁺.

Step 2. Synthesis of2S,4S,6S)-2'-chloro-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one (S33)

To a mixture of l-[(2^,S,6'S,7S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone (C154) (4.5 g, 10.24 mmol) in acetonitrile (70 mL) was added N-hydroxyphthalimide (1.2 g, 7.36 mmol) and cobaltous diacetate tetrahydrate (550 mg, 0.216 mmol), and then the mixture was vacuum purged

236 with an oxygen balloon three times. The mixture was heated to 45 °C and stirred for 18 hours before cooling to room température. The reaction was diluted with DCM, water, and saturated sodium bicarbonate, then extracted with DCM (3 x 150 mL) and collected through a phase separator. The organic layer was dried with NaiSCL, fîltered, and concentrated. Purification by silica gel chromatography (Gradient: 0-50% EtOAc in heptane) afforded (2S,4S,6S)-2'-chloro-2methyl-6-(l-rnethyltriazol-4-yl)-l-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3c]pyran]-4'-one S33 (3.50 g, 68%). Ή NMR (300 MHz, Chloroform-d) δ 7.61 (s, 1H), 7.19 (s, 1H), 5.61 (s, 1H), 4.44 (q, J = 7.1 Hz, 1H), 4.31 (s, 2H), 4.12 (s, 3H), 3.34 (dd, J = 15.1, 6.2 Hz, 1H), 2.78 (dd, J = 15.1, 8.3 Hz, 1H), 2.70 - 2.43 (m, 1H), 2.16 (s, 1H), 1.27 (d, J = 7.3 Hz, 3H). LCMS m/z 449.12 [M+H]⁺.

Compound 174 (2'S,4S,6'S, 7S)-2-chloro-2'-methyl-6’-(l-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-4-ol (174)

Step 1. Synthesis of 1-[(2S,4S,6'S, 7S)-2-chloro-4-hydroxy-2'-methyl-6'-(l-methyltriazol-4yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone (C63)

To (2S,4S,6S)-2'-chloro-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S33 (3.5 g, 7.025 mmol) in

237

DCM (60 mL) was added a solution of 1,2,3,4,5 pentamethylcyclopentane rhodium(2+) tetrachloride (24 mg, 0.03821 mmol) and N-[(lR,2R)-2-amino-l,2-diphenyl-ethyl]-4-methylbenzenesulfonamide (27 mg, 0.074 mmol) in DCM (7 mL) followed by a solution of formic acid (1.4 mL, 37.11 mmol) and triethylamine (2.1 mL, 15.07 mmol). The flask was fitted with an empty balloon to capture the CO₂ off-gas byproduct. After two hours, the mixture was washed with saturated aqueous sodium bicarbonate (150 mL). The organic phase was separated, passed through a phase separator, and concentrated. Silica gel purification (Column: 120 g silica gel, Gradient: 0-45% EtOAc in Heptane) afforded l-[(2'S,4S,6'S,7S)-2-chloro-4-hydroxy-2'-methyl6'-(l-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-T-yl]-2,2,2trifluoro-ethanone C63 (3.3 g, 86%) as a pale off-white foam. A NMR (300 MHz, Chloroformd) δ 7.59 (s, 1H), 6.83 (s, 1H), 5.53 (s, 1H), 4.46 (dt, J= 9.1, 3.1 Hz, 2H), 4.10 (s, 3H), 4.03 3.80 (m, 2H), 3.10 (dd, J= 15.1, 7.3 Hz, 1H), 2.65 (ddd, J= 15.1, 8.1, 2.2 Hz, 1H), 2.47 (s, 1H), 2.21 - 2.08 (m, 1H), 2.08 (d, J= 9.2 Hz, 1H), 1.40 - 1.19 (m, 3H). LCMS m/z 451.05 [M+H]⁺.

Note that stéréochemistry of alcohol C63 was assigned using NMR NOE studies and literature understanding of réductions using this catalyst and ligand System. (Reference: New Chiral Rhodium and Iridium Complexes with Chiral Diamine Ligands for Asymmetric Transfer Hydrogénation of Aromatic Ketones. Kunihiko Murata, Takao Ikariya, and Ryoji Noyori. The Journal of Organic Chemistry 1999 64 (7), 2186-2187).

Step 2. Synthesis of (2'S,4S,6'S,7S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol (174)

To a solution of l-[(2'S,4S,6'S,7S)-2-chloro-4-hydroxy-2'-methyl-6'-(l-methyltriazol-4yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4^,-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C63 (3.33 g, 100%) in MeOH (50 mL) was added NaOH (40 mL of 2 M, 80.00 mmol) and the mixture was stirred at 60 °C. After 40 minutes, the mixture was diluted with saturated aqueous ammonium chloride until pH 10 (about 50 mL) and extracted with MTBE (5 x 100 mL) and ethyl acetate (1 x 75 mL). The combined organic layers were washed with saturated aqueous NaCl, dried over Na₂SO4, and concentrated. The residue was brought up in EtOH and stripped down (3x) to afford a white solid. The solid was transferred to a vial and dried under vacuum at 55 °C ovemight to give amorphous (2'S,4S,6'S,7S)-2-chloro-2^,-methyl-6'-(l -methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol 174 (2.1817 g, 87%). *HNMR (400 MHz, Methanol-ck) δ 7.82 (s, 1H), 6.88 (s, 1H), 4.46 (t, 7= 3.8 Hz, 1H), 4.34 - 4.28 (m, 1H), 4.08 (s, 3H), 4.04 (dd, 7= 12.2, 3.6 Hz, 1H), 3.81 (dd, 7= 12.2, 4.1 Hz, 1H), 3.36 - 3.25 (m, 1H), 2.39 (dt, 7= 13.8, 2.6 Hz, 1H), 2.17 (dt, 7= 13.7, 2.6 Hz, 1H), 1.71 (dd,7= 13.9, 11.9 Hz, 1H), 1.45 (dd, 7= 13.7, 11.4 Hz, 1 H), 1.16 (d, 7= 6.4 Hz, 3H). LCMS m/z 355.03 [M+H]⁺.

238

Compounds 175 and 176 (2S)-2-chloro-2 '-methyl-6'-(l-methylimidazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4 piperidine]-4-ol (175)[DIASTERIOMER-1] and (176) [DIASTERIOMER-2]

NaOH

OH OH

[DIAST-1] [DIAST-2]

Step 1. Synthesis of l-[(2S)-2-chloro-4-hydroxy-2'-methyl-6'-(l-methylpyrazol-4yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4 '-piperidine]-1 '-yl]-2,2,2-trifluoro-ethanone (C64)[DIAST-1] and (C65)[DIAST-2] (7î)-(+)-2-Methyl-CBS-oxazaborolidine solution (40 pL of 1 M, 0.04000 mmol) (1 M solution in THF) in THF (1 mL) was cooled to 0 °C and treated with borane; tetrahydrofuran (220 pL of 1 M, 0.2200 mmol). After 4 minutes, a solution of (2S,4S,6S)-2'-chloro-2-methyl-6(l-methylpyrazol-4-yl)-l-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S35 (50 mg, 0.1023 mmol) in THF (300 pL) was added slowly and the reaction was stirred at 0 °C. After 15 minutes, another solution of (Æ)-(+)-2-Methyl-CBS-oxazaborolidine solution (40 pL of 1 M, 0.04000 mmol) and borane;tetrahydrofuran (220 pL of 1 M, 0.2200 mmol) was made and added to the reaction. After 30 minutes, the reaction was quenched with 2 N HCl, the ice bath was removed, and the mixture was stirred vigorously for 24 hours. The reaction was extracted with DCM (3x) through a phase separator. The organics were concentrated via rotovap. Purification by silica gel chromatography (0-60% EtOAc in Heptane) yielded racemic intermediate.

239

SFC séparation (AD-H column with 10% MeOH w/5 mM ammonia) gave 1-[(2S) 2chloro-4-hydroxy-2'-methyl-6'-(l-methylpyrazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-r-yl]-2,2,2-trifluoro-ethanone (C64) [DIAST-1] (50 mg, 101%) *H NMR (300 MHz, Chloroformé) δ 7.40 (d, J= 5.6 Hz, 2H), 6.88 (s, 1H), 5.52 (s, 1H), 4.52 - 4.37 (m, 2H), 4.00 - 3.76 (m, 5H), 2.74 - 2.48 (m, 2H), 2.27 (s, 1H), 1.81 (dd, 7= 14.9, 6.8 Hz, 1H), 1.28 (s, 3H). LCMS m/z 450.07 [M+H]⁺; l-[(2S)-2-chloro-4-hydroxy-2'-methyl-6'-(l-methylpyrazol-4yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone (C65) [DIAST-2] (34 mg, 62%) ‘H NMR (300 MHz, Chloroformé) δ 7.43 (s, 1H), 7.35 (s, 1H), 6.85 (s, 1H), 5.43 (s, 1H), 4.56 - 4.40 (m, 2H), 4.02 - 3.79 (m, 5H), 2.71 (ddd, J= 15.1, 8.1, 2.3 Hz, 1H), 2.37 (dd, J= 15.1, 6.4 Hz, 1H), 2.08 (d, 7= 9.2 Hz, 1H), 1.95 (dd, J= 14.6, 6.9 Hz, 1H), 1.27 (d, 7= 6.6 Hz, 3H). LCMS m/z 450.03 [M+H]⁺.

Step 2. Synthesis of (2S)-2-chloro-2'-methyl-6'-(l-methylimidazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol (175) [DIAST-1] and (176) [DIAST-1] l-[(2'S,4S,6'S,7S)-2-chloro-4-hydroxy-2'-methyl-6'-(l-methylpyrazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone (C64) (50 mg, 0.094 mmol) in MeOH (2 mL) was treated with NaOH (1 mL of 1 Μ, 1.000 mmol) and heated to 40 °C for 30 minutes. Additional NaOH (1 mL of 1 Μ, 1.000 mmol) was added and the reaction was heated to 50 °C. After two more hours the reaction was diluted with DCM. The organic phase was separated, passed through a phase separator, and concentrated. The material was brought up in MTBE (3 mL) and treated with hydrogen chloride (28 pL of 4 M, 0.1120 mmol) in dioxane dropwise. A white precipitate formed. The solution was concentrated and the residue brought up in water, frozen at -78 °C, and lyophilized over the weekend to afford (2S)-2-chloro-2'-methyl6'-(l-methylpyrazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol (Hydrochloride sait) (175) [DIAST-1] (37.9 mg, 98%). Ή NMR (400 MHz, Methanolé?) δ 7.72 (s, 1H), 7.58 (s, 1H), 6.92 (s, 1H), 4.52 - 4.45 (m, 2H), 4.03 (dd, 7= 12.2, 3.5 Hz, 1H), 3.88 (s, 3H), 3.83 (dd, 7= 12.2, 4.0 Hz, 1H), 3.69 - 3.58 (m, 1H), 2.47 (dt, 7= 14.5, 2.8 Hz, 1H), 2.33 (dt, 7= 14.4, 2.8 Hz, 1H), 1.98 (t,7= 13.7 Hz, 1H), 1.71 (t,7= 13.1 Hz, 1H), 1.30 (d, 7= 6.6 Hz, 3H). LCMS m/z 353.99 [M+H]⁺.

l-[(2'S,4R,6'S,7S)-2-chloro-4-hydroxy-2'-methyl-6'-(l-methylpyrazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone (C65) (34 mg, 0.07 mmol) in MeOH (1.5 mL) was treated with NaOH (800 pL of 1 M, 0.8 mmol) and heated to 40 °C for 30 minutes. More NaOH (800 pL of 1 M, 0.8 mmol) was added and the reaction was heated to 50 °C. After two more hours the reaction was cooled to room température, diluted with DCM. The organic phase was separated, passed through a phase separator, and concentrated. The 240 material was brought up in MTBE (1.5 mL) and treated with HCl (22 pL of 4 M, 0.08800 mmol) dropwise. A white precipitate formed. The solution was concentrated via rotovap and the residue brought up in water, frozen at -78 °C, and lyophilized ovemight to give (2S)-2-chloro-2'-methyl6'-(l-methylpyrazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol (Hydrochloride sait) (176) [DIAST-2] (26.7 mg, 96%) was afforded as a white powder. *H NMR (400 MHz, Methanol-^) δ 7.81 (s, 1H), 7.63 (s, 1H), 6.94 (s, 1H), 4.71 (dd, J= 12.6, 2.9 Hz, 1H), 4.50 (t, J= 3.6 Hz, 1H), 4.05 (dd, J= 12.3, 3.4 Hz, 1H), 3.90 (s, 3H), 3.85 (dd, J= 12.2, 3.9 Hz, 1H), 3.71 (dtq, J= 13.4, 6.8, 2.9 Hz, 1H), 2.53 (dt, J= 14.4, 2.8 Hz, 1H), 2.42 (dt, J= 14.8, 2.8 Hz, 1H), 2.21 (dd, 7=14.4,12.6 Hz, 1H), 1.72 (dd,7=14.8, 12.2 Hz, 1H), 1.35 (d, 7= 6.6 Hz, 3H). LCMS m/z 354.04 [M+H]⁺.

Compound 177 (2 'S, 4R, 6’S, 7S)-2-chloro-2 ’-methyl-6'-(l-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran7,4'-piperidine]-4-ol] (177)

Step 1. Synthesis of (2'S,4R,6'S,7S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol] (177)

To tetrahydrofuran (500 pL) cooled to 0 °C was added (3aR)-l-methyl-3,3-diphenyl3a,4,5,6-tetrahydropyrrolo[l,2-c][l,3,2]oxazaborole ((R)-CBS catalyst) (25 pLof 1 M, 0.025 mmol) followed by borane tetrahydrofuran (250 pL of 1 M, 0.25 mmol). After stirring for 5 minutes, a solution of (2S,4S,6S)-2'-chloro-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S33 (25 mg, 0.05570 mmol) in THF (1000 pL) was added, dropwise. The mixture stirred at 0 °C for 1 hour. The mixture was concentrated, diluted with MeOH (1.5 mL) and quenched with NaOH (100 pL of 6 M, 0.6000 mmol). The mixture was warmed to 50 °C and stirred ovemight. Purification by reversed-phase HPLC (Method: Cl8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 5 mM HCl) afforded (2'S,4R,6'S,7S)-2-chloro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[4,5241 dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol (Hydrochloride sait) 177 (3.0 mg, 13%). *H NMR (300 MHz, Methanol-70 δ 8.11 (d, J= 2.4 Hz, 1H), 6.94 (s, 1H), 4.90 (d, J= 3.1 Hz, 1H), 4.51 (t, J= 3.6 Hz, 1H), 4.13 (d, J= 2.2 Hz, 3H), 4.07 (dd, J= 12.2, 3.5 Hz, 1H), 3.87 (dd, J = 12.3, 3.9 Hz, 1H), 3.74 (ddt, J= 13.0, 9.4, 6.4 Hz, 1H), 2.78 - 2.51 (m, 1H), 2.51 - 2.19 (m, 2H), 5 1.87 (ddd, 7= 30.8, 14.7, 12.2 Hz, 1H), 1.39 (dd, 7= 6.6, 3.1 Hz, 3H). LCMS m/z 355.03

[M+H]⁺.

Compounds 178-182

Compounds 178-182 (see Table 5) were prepared in two or three steps from ketone intermediates in Table 5 using the réduction and deprotection methods as described for compounds 174-177. Final compounds were made through hydrolysis with NaOH. Any modifications to these methods are noted in Table 5 and accompanying footnotes.

Table 5. Method of préparation, structure and physicochemical data for compounds 178-182

Product	Ketone intermediate	Réduction method	¹H NMR; LCMS m/z [M+H]⁺
Compound 178 OH 1* ojCVci N //Ar n \ IJ H /	0 ” A S34	Compound 175 and 176⁶·⁷	'H NMR (300 MHz, Chloroform-7) δ 7.31 (s, 1H), 6.82 (s, 1H), 6.73 (s, 1H), 4.45 (t, 7=3.2 Hz, 1H), 4.23 (dd, 7= 11.6, 2.6 Hz, 1H), 4.01 (dd, 7= 12.2, 3.1 Hz, 1H), 3.88 (dd, 7=12.2, 3.4 Hz, 1 H), 3.62 (s, 3H), 3.22 (dtt, 7=12.5, 6.1,3.2 Hz, 1H), 2.23 (dt, 7 = 13.4, 2.6 Hz, 1H), 2.10 (dt, 7= 13.9, 2.6 Hz, 1H), 1.83 (dd,7 = 13.4, 11.6 Hz, 1H), 1.30 (dd, 7= 14.0,11.4 Hz, 1H), 1.11 (d, 7=6.4 Hz, 3H). LCMS m/z 354.04 [M+H]⁺

242

Compound 179 OH JL* nlV^cl ΝχΑ,Λ, <'J H ' lé /	0 °ÔQ-^ci ^</NJaS /ⁿ S34	Compound 175 and 176⁶·⁷	Ή NMR (300 MHz, Chloroformé δ 7.34 (s, 1H), 6.81 (s, 1H), 6.75 (s, 1H), 4.42 (t, 7= 2.7 Hz, 1H), 4.14 (dd, 7 = 11.8, 2.5 Hz, 1H), 4.03 (dd, 7= 12.4, 2.8 Hz, 1 H), 3.92 (dd, 7=12.4, 2.7 Hz, 1H), 3.62 (s, 3H), 3.44-3.26 (m, 1H), 2.39 (dd, 7= 13.9, 2.8 Hz, 1H), 2.06-1.96 (m, 1H), 1.68 (dd, 7 = 13.9, 11.7 Hz, 1H), 1.49 (dd, 7= 13.4, 11.3 Hz, 1H), 1.13 (d, J =6.3 Hz, 3H). LCMS m/z 354.04 [M+H]⁺
Compound 180 OH o Jé-u Jé F é/LA. N'J ü é /	0 é S32	Compound 177⁴·⁵	’H NMR (300 MHz, Methanolé) δ 7.89 (d, 7= 4.5 Hz, 1H), 7.47 (d, 7=1.3 Hz, 1H), 4.59 (t, 7= 3.8 Hz, 1H), 4.45 (dd, 7= 12.0,2.7 Hz, 1H), 4.13-4.02 (m, 4H), 3.87 (dd, 7=12.3, 4.1 Hz, 1H), 3.51 -3.34 (m, 1H), 2.47 (dt,7 = 14.0, 2.7 Hz, 1H), 2.27 (dt, 7= 13.8, 2.7 Hz, 1H), 1.95 (ddd, 7= 34.7, 13.9, 12.0 Hz, 1H), 1.57 (ddd, 7= 29.0, 14.0, 11.6 Hz, 1H), 1.21 (dd,7 = 6.5, 4.0 Hz, 3H). LCMS m/z 388.87 [M+H]⁺
Compound 181 OH Ν'-J) B é / (Note: Amorphous form obtained)	0 oÔCM-f ~X^S F Υχ; ' é S32	Compound I/4I.2J	Ή NMR (400 MHz, Methanolé) δ 7.83 (s, 1H), 7.46 (d, 7=1.2 Hz, 1H), 4.58 (t, 7= 3.8 Hz, 1H), 4.34 (dd, 7= 11.8, 2.6 Hz, 1H), 4.11-4.03 (m, 5H), 3.86 (dd,7= 12.2, 4.2 Hz, 1H), 3.393.34 (m, 1H), 2.46-2.41 (m, 1H), 2.26-2.19 (m, 1H), 1.78 (dd,7= 13.8, 11.9 Hz, 1H), 1.53 (dd,7 = 13.6, 11.5 Hz, 1H), 1.17 (d, 7= 6.5 Hz, 3H).

243

			LCMS m/z 389.14 [M+H]⁺.
Compound 182 OH ^ciajQ N X A Nⁿ	O ^CIAJ[ i -/V y nN S36	Compound 1743,2,8,9,10	Ή NMR (300 MHz, Chloroform-7) δ 7.43 (s, 1H), 6.63 (s, 1H), 4.53 (t, 7=2.9 Hz, 1H), 4.37 (dd, 7=11.8, 2.6 Hz, 1H), 4.05 (s, 3H), 3.99 (dd, 7= 13.8, 2.8 Hz, 2H), 3.35 (ddt, 7= 12.7, 6.4,3.2 Hz, 1H), 2.25 (dt, 7 = 13.9, 2.6 Hz, 1H), 1.86 (dt, 7=13.5, 2.5 Hz, 1H), 1.69 (dd,7 = 13.9, 11.8 Hz, 1H), 1.54 (dd, 7= 13.5, 11.3 Hz, 1H), 1.12 (d, 7= 6.4 Hz, 3H). LCMS m/z 355,08 [M+H]⁺.

1. Formic acid and triethylamine were added before Ketone intermediate (step 1)

2. Silica gel purification (0-60% EtOAc in Heptane) afforded product (step 1)

3. Product was not washed with EtOH (step 2)

4. Reaction was stirred for 15 minutes before quenching

5. Stirred at 40 °C for four hours after quenching with NaOH and MeOH (step 2)

6. Step 1 was run with both the CBS-(S) catalyst and the CBS-(R) catalysts in two separate reactions. However, both reactions proceeded with poor d.r. and so they were combined to make the racemic which was separated by SFC as in Step 2.

7. After extracted with DCM, purification by silica gel chromatography (Gradient: 0-20%

MeOH in DCM) yielded the product (step 3)

8. Reaction was stirred overnight (stepl)

9. Stirred at 50 °C for two hours after quenching with NaOH and MeOH (step 2)

10. Compound 182 contained approximately 15% of a diastereomer created via différentiation from the enantiomer of S36 présent in S36.

244

Compound 181 (2S, 4S, 4 'S, 6S)-2-methyl-6-(l -methyl-lH-1,2,3-triazol-4-yl)-2 '-(trifluoromethyl)-4 5 dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-ol (181), Amorphous Form

Cp*RhCI[(R,R)-Tsdpen]

HCO₂H, NEt₃

OH

F

S32

C153

NaOH

OH

181

N H

Step 1. Synthesis of2,2,2-trifluoro-l-[(2'S,4S,6'S,7S)-4-hydroxy-2^,-methyl-6'-(lmethyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine] l'-yl]ethenone (C153)

To (2S,4S,6S)-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2-trifluoroacetyl)-2'(trifluoromethyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S32 (2.23 g, 4.63 mmol) in DCM (20 mL) was added a solution of 1,2,3,4,5 pentamethylcyclopentane rhodium(2+) tetrachloride (7 mg, 0.002 mmol) and N-[(lR,2R)-2-amino-l,2-diphenyl-ethyl]-4-methylbenzenesulfonamide (8.5 mg, 0.005 mmol) in DCM (2 mL), followed by a solution of formic acid (0.9 mL, 5.15 mmol) and triethylamine (1.3 mL, 2.01 mmol). The flask was fitted with an empty balloon to capture the CO₂ off-gas byproduct. Afiter three hours, the mixture was washed with saturated aqueous sodium bicarbonate (10 mL). The organic phase was separated, passed through a phase separator, and concentrated. Silica gel purification (Column: 40 g silica gel, Gradient: 0-50% EtOAc in Heptane) afforded 2,2,2-trifluoro-l-[(2'S,4S,6'S,7S)-4-hydroxy-2'methyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'245 piperidine]-r-yl]ethenone C153 (2.27 g, 100%) as a white solid. LCMS m/z 485.11 [M+H]⁺. Step 2. Synthesis of (2'S,4S,6'S,7S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4’-piperidine]-4-ol (181)

To a solution of2,2,2-trifluoro-l-[(2'S,4S,6'S,7S)-4-hydroxy-2'-methyl-6'-(lmethyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-ryl]ethenone C153 (2.27 g, 4.63 mmol) in MeOH (45 mL) was added NaOH (8 mL of 6 M, 48.00 mmol), and the mixture was stirred at 60 °C. After 75 minutes, the mixture was diluted with saturated aqueous ammonium chloride until pH 10 (about 40 mL), water (40 mL), and extracted with MTBE (100 mL). The aqueous layer was extracted with additional MTBE (2 x 50 mL), and the combined organic layers were washed with saturated aqueous NaCl, dried over MgSCL, and concentrated to give amorphous (2'S,4S,6'S,7S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol 181 (1.84 g, 88%). *H NMR (400 MHz, Chloroform-d) δ 7.48 (s, 1H), 7.39 (q, J = 1.2 Hz, 1H), 4.58 (d, J = 8.0 Hz, 1H), 4.44 (dd, J = 11.7, 2.5 Hz, 1H), 4.09 (s, 4H), 4.01 (dd, J = 12.5, 2.7 Hz, 1H), 3.43 (ddd, J = 11.2, 6.4, 2.5 Hz, 1H), 2.48 (dt, J = 13.8, 2.6 Hz, 1H), 2.16 - 2.07 (m, 2H), 1.77 (dd, J = 13.9, 11.8 Hz, 1H), 1.63 (s, 1H), 1.28 (s, 1H), 1.18 (d, J = 6.3 Hz, 3H). LCMS m/z 389.09 [M+H]⁺·

Compound 183 (2 'S, 6'S, 7S)-2-chloro-4,4-difluoro-2 '-methyl-6'-(l-methyltriazol-4-yl)spiro[5H-thieno[2,3c]pyran-7,4'-piperidine] (183)

S33 C66

Step 1. Synthesis of l-[(2'S,6'S,7S)-2-chloro-4,4-difluoro-2'-methyl-6'-(l-methyltriazol-4yl)spiro[5H-thieno[2,3-c]pyran- 7,4 ’-piperidine]-1 ’-yl]-2,2,2-trifluoro-ethanone (C66) (2S,4S,6S)-2'-chloro-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S33 (40 mg, 0.08912 mmol) was dissolved in DCM (100 pL) and DAST (35 pL, 0.2649 mmol) was added. The reaction was heated to 40 °C and stirred for 3 hours. Additional DAST (35 pL, 0.2649 mmol) was added and stirred over the weekend. The solution was diluted with DCM and poured into aqueous NaHCCh

246 stirring at 0 °C. The organic phase was separated, passed through a phase separator, and concentrated to give crude l-[(2'S,6'S,7S)-2-chloro-4,4-difluoro-2'-methyl-6'-(l-methyltriazol-4yl)spiro[5H-thieno[2,3-c]pyran-7,4'-piperidine]-T-yl]-2,2,2-trifluoro-ethanone C66 (41 mg, 42%). LCMS m/z 468.71 [M+H]⁺.

Step 2. Synthesis of (2'S,6'S,7S)-2-chloro-4,4-difluoro-2'-methyl-6'-(l-methyltriazol-4yl)spiro[5H-thieno[2,3-c]pyran- 7,4 ’-piperidine] (183)

[(2'S,6'S,7S)-2-chloro-4,4-difluoro-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[5Hthieno[2,3-c]pyran-7,4'-piperidine]-T-yl]-2,2,2-trifluoro-ethanone C66 was stirred in 6M aqueous NaOH (10 eq) for five hours. Solvent was removed through rotary évaporation. Purification by reversed-phase HP LC (Method: Cl 8 Waters Sunfire column (30 x 150 mm, 5 micron) with 10 mM Ammonium Hydroxide) to afford (2'S,6'S,7S)-2-chloro-4,4-difluoro-2'methyl-6'-(l-methyltriazol-4-yl)spiro[5H-thieno[2,3-c]pyran-7,4'-piperidine] 183 (2 mg, 6%). LCMS m/z 375.11 [M+H]⁺.

Compounds 184 and 185 (2'S,6'S, 7S)-2-chloro-4-(difluoromethyl)-2'-methyl-6'-(l-methyltriazol-4-yl)spiro[5H-thieno[2,3c]pyran-7,4'-piperidine]-4-ol (184) [DIAST-1] and (185) [DIAST-2]

DIAST-1 DIAST-2

To a solution of (2S,4S,6S)-2'-chloro-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S33 (65 mg, 0.1448 mmol) in MeCN (2.4 mL) was added DMPU (34 pL, 0.2822 mmol) followed by [bromo(difluoro)methyl]trimethyl-silane (35 mg, 0.1723 mmol) and PPh₃ (40 pL, 0.1726 mmol). The resulting solution was heated at 55 °C. After four hours the reaction was cooled to room température. Aqueous KOH (720 pL of 1 M, 0.7200 mmol) was added and the reaction was stirred for 48 hours. The reaction was quenched with sodium bicarbonate and DCM. The organic phase was separated, passed through a phase separator. Purification by reversed-phase HP LC (Method: Cl 8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 0.1% trifluoroacetic) was performed.

247

Some impurities still existed. Purification by silica gel chromatography (Gradient: 0-20% MeOH in DCM) yielded both separated diastereomers (2'S,6'S,7S)-2-chloro-4-(difluoromethyl)2'-methyl-6'-(l-methyltriazol-4-yl)spiro[5H-thieno[2,3-c]pyran-7,4'-piperidine]-4-ol (Trifluoroacetate sait) 184 [DIAST-1] (4.3 mg, 6%). ^JH NMR (400 MHz, Methanol-^) δ 7.94 (s, 1H), 7.01 (s, 1H), 5.97 (t, J= 55.2 Hz, 1H), 4.57 (dd, J= 12.3, 2.9 Hz, 1H), 4.10 (s, 4H), 3.81 (dt, J= 12.6, 2.2 Hz, 1H), 3.60 - 3.42 (m, 1H), 2.60 (dt, J= 14.4, 2.7 Hz, 1H), 2.18 (dt, J= 14.1, 2.6 Hz, 1H), 2.08 - 1.95 (m, 1H), 1.69 (dd, J = 14.2, 11.8 Hz, 1H), 1.26 (d, .7= 6.5 Hz, 3H). LCMS m/z 405.02 [M+H]⁺. (2'S,6'S,7S)-2-chloro-4-(difluoromethyl)-2'-methyl-6'-(lmethyltriazol-4-yl)spiro[5H-thieno[2,3-c]pyran-7,4'-piperidine]-4-ol (Trifluoroacetate sait) 185 [DIAST-2] (8.5 mg, 11%). *H NMR (400 MHz, Methanol-^) δ 8.06 (s, 1H), 7.04 (d, J= 0.9 Hz, 1H), 5.98 (t, J= 55.1 Hz, 1H), 4.91 - 4.86 (m, 1H), 4.20 - 4.05 (m, 4H), 3.90 - 3.71 (m, 2H), 2.64 - 2.33 (m, 3H), 1.85 (dd, J= 14.8, 12.2 Hz, 1H), 1.40 (d, J= 6.6 Hz, 3H). LCMS m/z 405.02 [M+H]⁺.

Préparation ofS37

4-aminohexan-2-one hydrochloride sait (S3 7)

Step 1. Synthesis of tert-butyl N-[l-(p-tolylsulfonyl)propyl]carbamate (C68)

To a solution of tert-butyl carbamate C67 (20 g, 0.1639 mol) and 4methylbenzenesulfmate (53 g, 0.3313 mol) in MeOH (120 mL) and H₂O (240 mL) was added propanai (15.082 g, 19 mL, 0.2545 mol) and formic acid (76.921 g, 65 mL, 1.6211 mol) dropwise at 0 °C. The reaction mixture was stirred at room température for 24 hours. The reaction mixture was filtered, washed with water (100 mL), and dried to afford teri-butyl N-[l(p-tolylsulfonyl)propyl]carbamate C68 (46 g, 85%). ^rH NMR (300 MHz, DMSO-î/ô) δ 7.82 (d, J

248 = 9.6 Hz, 1H), 7.65 (d, J= 8.4 Hz, 2H), 7.48-7.40 (m, 2H), 4.63-4.56 (m, 1H), 2.39 (d, J= 14.4 Hz, 3H), 2.01-1.95 (m, 1H), 1.69-1.61 (m, 1H), 1.17 (s, 9H), 0.91 (t,7=7.2 Hz, 3H) as an off white solid.

Step 2. Synthesis of tert-butyl 2-acetyl-3-(tert-butoxycarbonylamino) pentanoate (C69) To a solution of NaH (2.5 g, 60% (w/w), 0.0625 mol) in THF (160 mL) was added tertbutyl N-[l-(p-tolylsulfonyl)propyl]carbamate C68 (10 g, 0.0303 mol) portion-wise at room température and stirred for 5 minutes and then teri-butyl 3-oxobutanoate (5.1001 g, 5.4 mL, 0.0319 mol) in THF (40 mL) was added dropwise and stirred at same température for 2 hours. The reaction mixture was quenched with NH₄C1 solution (150 mL) and extracted with DCM (2 x 300 mL). The organic layer was dried over anhydrous Na2SÜ4 concentrated under reduced pressure and dried to provide crude ter/-butyl 2-acetyl-3-(tert-butoxycarbonylamino)pentanoate C69 (11 g, 98%) as a yellow gum. Ή NMR (300 MHz, DMSO-70 δ 6.72-6.56 (m, 1H), 3.973.89 (m, 1H), 3.58-3.44 (m, 1H), 2.16 (s, 3H), 1.48-1.15 (m, 20H), 0.85-0.76 (m, 3H). LCMS m/z 316.27 [M+H]⁺.

Step 3. Synthesis of 4-aminohexan-2-one (S37)

A solution of ZerZ-butyl 2-acetyl-3-(tert-butoxycarbonylamino)pentanoate C69 (11 g, 0.0296 mol) in 10% aqueous HCl (120 mL of 10% (w/v), 0.3291 mol) was heated to 110 °C and stirred for 2 hours. The reaction mixture was extracted with diethyl ether (4 x 50 mL). The aqueous part was evaporated under reduced pressure and dried to afford 4-aminohexan-2-one S37 (Hydrochloride Sait) (3.92 g, 85%) as a pale brown liquid. Ή NMR (400 MHz, DMSO-70 δ 8.00 (s, 3H), 3.38-3.30 (m, 1H), 2.89-2.73 (m, 2H), 2.14 (s, 3H), 1.63-1.49 (m, 2H), 0.91 (t, 7 = 10.4 Hz, 3H). LCMS m/z 116.3 [M+H]⁺.

249

Préparation of S38

4-amino-5-methyl-hexan-2-one hydrochloride sait (S38)

Step 1. Synthesis of tert-butyl N-[2-methyl-l-(p-tolylsulfonyl)propyl] carbamate (C71)

To a stirred solution of 2-methylpropanal (18.652 g, 23.610 mL, 0.2535 mol) and tertbutyl carbamate C70 (20 g, 0.1690 mol) in MeOH (200 mL) and Water (50 mL) at room température was added sodium79-Îoluenesulfmate (60.835 g, 0.3380 mol) followed by formic acid (79.370 g, 65.057 mL, 1.6900 mol) and then the reaction mixture was stirred at room température for 24 hours. The reaction mixture was filtered, washed with water (200 mL) and diethyl ether (50 mL), and dried under vacuum to afford crude tert-butyl N-[2-methyl-l-(ptolylsulfonyl)propyl]carbamate C71 (45 g, 73%) as an off white solid. Ή NMR (300 MHz, DMSO-î/ô) δ 7.81 (d, J= 10.5 Hz, 1H), 7.67 (d, J= 8.1 Hz, 2H), 7.39 (d, J= 8.1 Hz, 2H), 4.58 (q, J= 5.7 Hz, 1H), 2.46-2.36 (m, 4H), 1.14 (s, 9H), 1.00 (s, 6H).

Step 2. Synthesis of tert-butyl 2-acetyl-3-(tert-butoxycarbonylamino)-4-methyl-pentanoate (C72)

To a stirred solution of NaH (2.1998 g, 0.0550 mol) in THF (100.00 mL) was added tertbutyl N-[2-methyl-l-(p-tolylsulfonyl)propyl]carbamate C71 (10 g, 0.0275 mol) at room température and the reaction was stirred for 20 minutes. tert-Butyl 3-oxobutanoate (5.3270 g, 5.6074 mL, 0.0330 mol) in THF (30.000 mL) was added and stirred for 2 hours at room température. The reaction mixture was poured into saturated ammonium chloride (500 mL) at 10 °C and extracted with EtOAc (2 x 200 mL). The combined organic layers were dried over anhydrous Na₂SÛ4 and filtered, concentrated under reduced pressure to get crude tert-butyl 2acetyl-3-(tert-butoxycarbonylamino)-4-methyl-pentanoate C72 (11 g, 45%) as abrown liquid. LCMS m/z 330.2 [M+H]⁺.

250

Step 3. Synthesis of 4-amino-5-methyl-hexan-2-one (S38)

A solution of tert-butyl 2-acetyl-3-(tert-butoxycarbonylamino)-4-methyl-pentanoate C72 (15 g, 0.0319 mol) in HCl (290 mL of 10% (w/v), 0.7954 mol) was heated to 110 °C for 3 hours. The reaction mixture was concentrated under reduced pressure to obtained crude compound (6.2 g). The reaction was titrated with n-pentane (2 x 100 mL) and dried under vacuum to afford 4amino-5-methyl-hexan-2-one S38 (Hydrochloride Sait) (5.1 g, 96%), as abrown semi-solid. ’H NMR (400 MHz, DMSO-7₆) δ 7.96 (br s, 3H), 3.34-3.28 (m, 1H), 2.86-2.75 (m, 2H), 2.15 (s, 3H), 1.94-1.86 (m, 1H), 0.88 (s, 6H) LCMS m/z 130.2 [M+H]⁺.

Préparation of S39 l-cyclobutyl-3-oxo-butyl)ammonium chloride (S39)

Step 1. Synthesis of tert-butyl N-[cyclobutyl(p-tolylsulfonyl)methyl]carbamate (C74) To a mixture of sodiump-toluenesulfinate (27.843 g, 0.1547 mol) and tert-butyl carbamate C73 (Hydrochloride Sait) (12 g, 0.0773 mol) in MeOH (120.00 mL) and Water (240.00 mL) was added formic acid (12.20 g, 10.00 mL, 0.2598 mol) and cyclobutanecarbaldéhyde C77 (10 g, 0.1177 mol) at 0 °C. The mixture was stirred under nitrogen at room température for 48 hours. The white precipitate was filtered under vacuum, washed with water (3 x 50 mL), and dried to afford tert-butyl N-[cyclobutyl(ptolylsulfonyl)methyl]carbamate C74 (25 g, 93%), as a white solid. *H NMR (300 MHz, DMSOd₆) δ 7.82 (d, J= 9.9 Hz, 1H), 7.67 (d, J= 8.4 Hz, 2H), 7.42 (d, 7 = 8.1 Hz, 2H), 4.72 (t, J= 9.3 Hz, 1H), 2.86-2.82 (m, 1H), 2.37 (s, 3H), 2.01-1.73 (m, 6H), 1.18 (s, 9H).

251

Step 2. Synthesis of tert-butyl 2-[(tert-butoxycarbonylamino)-cyclobutyl-methyl]-3-oxobutanoate (C75)

A mixture of tert-butyl N-[cyclobutyl(p-tolylsulfonyl)methyl]carbamate C74 (25 g, 0.0722 mol) and ZerZ-butyl 3-oxobutanoate (Hydrochloride Sait) (15.5 g, 0.0788 mol) in THF (550 mL) was added NaH (5.35 g, 60% (w/w), 0.133 mmol) at 0 °C. The mixture was stirred under nitrogen at room température for 48 hours, and white precipitate was filtered under vacuum, washed with water (3x5 mL), and dried to afford tert-butyl 2-[(tertbutoxycarbonylamino)-cyclobutyl-methyl]-3-oxo-butanoate C75 (25 g, 99%), as pale yellow oil. Ή NMR (400 MHz, Chloroform-i/): δ 4.30-4.11 (m, 1H), 3.46 (d, J= 4.0 Hz, 1H), 2.49-2.42 (m, 1H), 2.25 (s, 3H), 1.94-1.74 (m, 6H), 1.48-1.44 (m, 18H). LCMS m/z 342.13 [M+l]⁺.

Step 3. Synthesis of (l-cyclobutyl-3-oxo-butyl)ammonium chloride (S39)

To a mixture of teri-butyl 2-[(tert-butoxycarbonylamino)-cyclobutyl-methyl]-3-oxobutanoate C75 (25 g, 0.0718 mol) in Water (100 mL) was added with HCl (100 mL of 5 M, 0.5000 mol) at room température. The mixture was stirred under nitrogen at 110 °C for 16 hours. The reaction mixture was diluted with water (100 mL) and washed with diethyl ether (3 x 100 mL). The aqueous layer was evaporated under vacuum and dried to afford (l-cyclobutyl-3-oxobutyl)ammonium chloride S39 (12.81 g, 100%), as pale brown semisolid. Ή NMR (400 MHz, DMSO-A) δ 7.86 (brs, 3H), 3.40-3.36 (m, 1H), 2.69-2.67 (m, 2H), 2.51-2.43 (m, 1H), 2.14 (s, 3H), 1.97-1.68 (m, 6H). LCMS m/z 142.2 [M+H]⁺.

Préparation of S40

4-amino-5-methoxy-pentan-2-one hydrochloride sait (S40)

252

Step 1. Synthesis of 2-methoxyacetaldehyde (C77)

A stirred solution of 1,1,2-trimethoxyethane C76 (20 g, 0.1631 mol) in aqueous HCl (300 mL of 0.5 M, 0.1500 mol) was heated to 55 °C for 4 hours. The reaction mixture was cooled to room température and saturated with excess NaCl sait. The reaction was extracted with DCM (5 x 500 mL). The combined organic layer was dried over sodium sulfate, fïltered, and evaporated under vacuum at below 25 °C to afford crude 2-methoxyacetaldehyde C77 (10 g, 66%) as a yellow liquid. ‘H NMR (400 MHz, Chloroform-7) δ: 9.73 (s, 1H), 4.05 (s, 2H), 3.52 (s, 3H).

Step 2. Synthesis of tert-butyl N-[2-methoxy-l-(p-tolylsulfonyl)ethyl] carbamate (C78)

To a stirred solution of Ze/7-butyl carbamate (2.2 g, 0.0186 mol) in MeOH (10 mL) and water (20 mL) were added sodiumy?-toluenesulfinate(6.7 g, 0.0368 mol), 2methoxyacetaldehyde C77 (2 g, 0.0243 mol) and formic acid (8.784 g, 7.20 mL, 0.1889 mol) at 0 °C. The reaction mixture was stirred at room température for 16 hours. The reaction mixture was extracted with DCM (3 x 100 ml). The combined organic extracts were dried over Na2SO₄, fïltered and evaporated under vacuum. Purification by silica chromatography (Eluent: 20% EtOAc in pet. ether) afforded tert-butyl N-[2-methoxy-l-(p-tolylsulfonyl)ethyl]carbamate C78 (5 g, 68%) as a white solid. Ή NMR (400 MHz, Chloroform-7) δ: 7.78 (d, J= 8 Hz, 2H), 7.32 (d, J = 8 Hz, 2H),4.99-4.94 (m, 1H), 4.13-4.09 (m, 1H), 3.83-3.79 (m, 1H), 3.42 (s, 3H),2.42 (s, 3H), 1.26 (s, 9H). LCMS m/z 330.36 [M+H]⁺.

Step 3. Synthesis of tert-butyl 2-acetyl-3-(tert-butoxycarbonylamino)-4-methoxy-butanoate (C79)

To a stirred solution of NaH (1 g, 57% (w/w), 0.0238 mol) in THF (35 mL) was added tert-butyl 3-oxobutanoate (2.35 g, 2.4737 mL, 0.0147 mol) at 0 °C. The reaction was stirred at 0 °C for 30 minutes. A solution of tert-butyl N-[2-methoxy-l-(p-tolylsulfonyl)ethyl]carbamate C78 (5 g, 0.0126 mol) in THF (35 mL) was added to the reaction dropwise at 0 °C. The reaction was allowed to stir for 3 hours at room température. The reaction mixture was quenched with ice cold water (100 mL) and extracted with EtOAc (2 x 150 mL). The combined organic extracts were dried over Na2SO₄, fïltered and evaporated under vacuum to afford terZ-butyl 2-acetyl-3(tert-butoxycarbonylamino)-4-methoxy-butanoate C79 (5 g, 72%) as a pale-yellow liquid. ^!H NMR (400 MHz, DMSO-ifc) δ: 5.55-5.41 (m, 1H), 3.78 (dd, J= 16.4 Hz, 1H), 3.49-3.45 (m, 1H), 3.37-3.30 (m, 4H), 2.25 (s, 3H), 1.49-1.41 (m, 18H).

Step 4. Synthesis of 4-amino-5-methoxy-pentan-2-one (S40)

A solution of tert-butyl 2-acetyl-3-(tert-butoxycarbonylamino)-4-methoxy-butanoate C79 (18 g, 0.0473 mol) in aqueous HCl (200 mL of 1 M, 0.2000 mol) was stirred at 55 °C for 3 hours. After cooling to room température, the mixture was washed with diethylether (2 x 100

253 mL). The aqueous layer was evaporated under vacuum to afford 4-amino-5-methoxy-pentan-2one S40 (Hydrochloride Sait) (7.4 g, 93%). Ή NMR (400 MHz, DMSOé₆) δ: 8.17 (brs, 3H), 3.64-3.54 (m, 1H), 3.47-3.40 (m, 2H), 3.28 (s, 3H), 2.83 (t, J= 6.4 Hz, 2H), 2.13 (s, 3H). LCMS m/z 132.2 [M+H]⁺.

Préparation of S41

2-ethyl-6-(l-methyltriazol-4-yl)piperidin-4-one (S41)

O

S41 \

[CIS]

4-aminohexan-2-one S37 (Hydrochloride sait) (298 mg, 1.965 mmol) was dissolved in EtOH (9 mL) and to it was added TEA (280 pL, 2.009 mmol), l-methyltriazole-4-carbaldehyde S17 (225 mg, 2.025 mmol), L-proline (47 mg, 0.4082 mmol) and MgSO₄ (255 mg, 2.119 mmol). The reaction was stirred at room température for 16 hours. The mixture was filtered, concentrated, quenched with saturated aqueous NaHCCh (50 mL), and extracted with DCM (5 x 20 mL). The combined organic layers were dried with MgSO₄, filtered, and concentrated under reduced pressure. Purification by silica gel chromatography (Gradient: 0-10% MeOH in DCM) afforded 2-ethyl-6-(l-methyltriazol-4-yl)piperidin-4-one S41 (162 mg, 38%). *H NMR (300 MHz, Chloroformé) δ 7.46 (d, J= 2.0 Hz, 1H), 4.22 (ddd, J= 10.3, 5.0, 2.0 Hz, 1H), 4.09 (d, J = 2.1 Hz, 3H), 2.93 (dtd, J= 11.8, 6.3, 2.9 Hz, 1H), 2.77 - 2.56 (m, 2H), 2.48 (ddd, J= 14.1, 2.9, 1.6 Hz, 1H), 2.16 (dd, 7= 14.1, 11.7 Hz, 1H), 1.74 - 1.50 (m, 2H), 0.98 (td,7= 7.6, 2.0 Hz, 3H). LCMS m/z 209.08 [M+H]⁺. Minor trans isomer purged during isolation.

Préparation of S42

2-isopropyl-6-(l-methyltriazol-4-yl)piperidin-4-one (S42)

O

S42 \

[CIS]

254

4-amino-5-methyl-hexan-2-one S38 (Hydrochloride sait) (500 mg, 3.018 mmol) was dissolved in EtOH (15 mL) and to it was added TEA (430 pL, 3.085 mmol), l-methyltriazole-4carbaldehyde S17 (350 mg, 3.150 mmol), L-proline (72 mg, 0.6254 mmol) and MgSCU (392 mg, 3.257 mmol) The reaction was stirred at room température for 3 days. The mixture was filtered, concentrated, and quenched with saturated aqueous bicarbonate (50 mL) and extracted with DCM (5 x 20 mL). The combined organic layers were dried with MgSCU, filtered, and concentrated. Crude ’H NMR showed about 5.5:1 dr. Purification by silica gel chromatography (Gradient: 0-5% MeOH in DCM) afforded =2-isopropyl-6-(l-methyltriazol-4-yl)piperidin-4-one S42 (272 mg, 38%). Ή NMR (300 MHz, Chloroform-7) δ 7.45 (s, 1H), 4.19 (dd, J= 11.0, 4.2 Hz, 1H), 4.10 (s, 3H), 2.79 (ddd, J= 11.8,5.7, 2.9 Hz, 1H), 2.73 - 2.54 (m, 2H), 2.46 (ddd, J = 13.9, 2.8, 1.7 Hz, 1H), 2.21 (dd, 7=13.9,11.8 Hz, 1H), 2.08 (s, 1H), 1.78 (dt,7=13.2, 6.6 Hz, 1H), 0.98 (t, 7= 6.5 Hz, 6H). The minor trans isomer was purged during isolation.

Préparation of S43

2-cyclobutyl-6-(l-methyltriazol-4-yl)piperidin-4-one (S43)

O

[CIS]

To a solution of 4-amino-4-cyclobutylbutan-2-one S39 (Hydrochloride sait) (1260 mg, 7.092 mmol) in EtOH (38 mL) was added TEA (1.0 mL, 7.175 mmol), l-methyltriazole-4carbaldehyde S17 (835 mg, 7.516 mmol), L-proline (168 mg, 1.459 mmol) and MgSO4(919 mg, 7.635 mmol). The reaction was stirred at room température for 24 hours. The mixture was filtered, concentrated, and quenched with saturated aqueous bicarbonate (50 mL) and extracted with DCM (5 x 20 mL). The combined organic layers were dried with MgSÛ4, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-3% MeOH in DCM) afforded, 2-cyclobutyl-6-(l-methyltriazol-4-yl)piperidin-4-one S43 (849 mg, 48%). ’H NMR (300 MHz, Chloroform-7) δ 7.45 (s, 1H), 4.27 - 4.16 (m, 1H), 4.09 (s, 3H), 2.92 (ddd, 7= 11.5, 8.6, 2.8 Hz, 1H), 2.67 - 2.56 (m, 2H), 2.49 - 2.30 (m, 2H), 2.23 - 1.70 (m, 8H). The minor trans isomer was purged during isolation.

255

Préparation of S44

2-(methoxymethyl)-6-(l-methyltriazol-4-yl)piperidin-4-one (S40)

O

To a solution of 4-amino-5-methoxy-pentan-2-one S40 (Hydrochloride sait) (750 mg, 4.474 mmol) in EtOH (22 mL) was added TEA (650 pL, 4.664 mmol), l-methyltriazole-4carbaldehyde S17 (520 mg, 4.680 mmol), L-proline (107 mg, 0.9294 mmol) and MgSCU (590 mg, 4.902 mmol). The reaction was stirred at room température for 16 hours. The mixture was filtered, concentrated, and quenched with saturated aqueous bicarbonate (50 mL) and extracted with DCM (7 x 20 mL). The combined organic layers were dried with sodium sulfate, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-12% MeOH in DCM) provided 2-(methoxymethyl)-6-(l-methyltriazol-4-yl)piperidin-4-one S44 (373 mg, 37%). 'H NMR (300 MHz, Chloroform-7) δ 7.47 (s, 1H), 4.24 (dd, J= 10.1, 5.0 Hz, 1H), 4.09 (s, 3H), 3.48 (dd, J = 9.2, 3.3 Hz, 1H), 3.43 - 3.34 (m, 4H), 3.25 (ddt, J = 10.9, 7.4, 3.8 Hz, 1H), 2.71 2.59 (m, 3H), 2.40 - 2.28 (m, 2H). LCMS m/z 225.06 [M+H]⁺. The product was isolated as a mixture of 5:1 dr.

Compound 186

2-chloro-2 ’-ethyl-6'-(l-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4'-piperidine] (186)

IVIOJ

[isomer 1 Et and Metriazole are CIS]

To amixture of 2-ethyl-6-(l-methyltriazol-4-yl)piperidin-4-one S41 (31 mg, 0.1414 mmol) and 2-(5-chloro-3-thienyl)ethanol (26 mg, 0.1599 mmol) S2 in DCM (700 pL) was added MsOH (37 pL, 0.5702 mmol) and the mixture was heated to 40 °C for 4 hours and then stirred at room température for 72 hours. The reaction was quenched with saturated NaHCCh solution and

256 extracted with DCM (6x). The combined organic layers were dried over anhydrous NaiSCU, fïltered, and concentrated under reduced pressure. Purification by silica gel chromatography (Gradient: 0-10% MeOH in DCM). The final product was dissolved in MeCN and water and subjected to lyophilization to provide 2-chloro-2'-ethyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] 186 (42.2 mg, 75%). *H NMR (300 MHz, Chloroformé δ 7.43 (s, 1H), 6.58 (s, 1H), 4.40 (dd, J= 11.8, 2.7 Hz, 1H), 4.06 (s, 3H), 4.00 3.89 (m, 2H), 3.15 - 2.99 (m, 1H), 2.74 - 2.49 (m, 2H), 2.36 (dt, J= 13.5, 2.6 Hz, 1H), 2.10 (dt, J = 13.7, 2.5 Hz, 1H), 1.96- 1.74 (m, 2H), 1.42 (qt, J= 11.1, 4.9 Hz, 3H), 0.94 (t, J= 7.6 Hz, 3H). LCMS m/z 353.05 [M+H]⁺.

Compound 187

2'-ethyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine] (187)

[isomer 2 Et and Metriazole are CIS]

To amixture of 2-ethyl-6-(l-methyltriazol-4-yl)piperidin-4-one S41 (31 mg, 0.1414 mmol) and 2-[5-(trifluoromethyl)-3-thienyl]ethanol (31 mg, 0.1580 mmol) S3 in DCM (700 pL) was added MeOH (37 pL, 0.5702 mmol) and the mixture was heated to 40 °C for 3 hours. To the solution was added MsOH (11 pL) and stirred at 40 °C for 1.5 hours and then stirred at 35 °C for 3.5 days. To the solution was added MsOH (11 pL) and stirred at 50 °C for 5 days during that time the solvent evaporated. The reaction was quenched with saturated NaHCCh solution and extracted with DCM (6x). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated. Purification by reversed-phase HP LC (Method: Cl 8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 0.1% trifluoroacetic acid) to provide 2^,-ethyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] (Trifluoroacetate sait) 187 (33.5 mg, 46%). ’H NMR (300 MHz, Chloroformé δ 7.89 (s, 1H), 7.16 (s, 1H), 4.94 (d, J= 10.2 Hz, 1H), 4.09 (s, 3H), 3.98 (h, J = 6.2 Hz, 2H), 3.67 (s, 1H), 2.93 - 2.64 (m, 3H), 2.49 - 2.34 (m, 2H), 2.19 - 2.04 (m, 2H), 1.94 (s, 1H), 1.67 (dt, J= 14.5, 7.8 Hz, 1H), 1.00 (t, J= 7.5 Hz, 3H). LCMS m/z 387.16 [M+H]⁺.

257

Compounds 188-193

Compounds 188-193 (see Table 6) were prepared in one step in an Oxa-Pictet Spengler reaction with thiophene éthanol intermediates S2 and S3 and piperidones (S42-S44). Any modifications to the methods are noted in Table 6 and accompanying footnotes.

Table 6. Method of préparation, structure, and physiochemical data for compounds 188-193

Product	Piperidone	Method	*H NMR; LCMS m/z [M+H]⁺
Compound 188 ^ci^CO> ^s pA 1 H ί ' ,N \	0 \| H t N S42 \ [GIS]	Compound 186 '·²·³	‘H NMR (300 MHz, Chloroform-7) δ 7.47 (s, 1H), 6.59 (s, 1H), 4.43 (dd, J= 11.9, 2.6 Hz, 1H), 4.06 (s, 3H), 3.95 (td, 7=5.4, 1.4 Hz, 2H), 2.95 (ddd, 7= 11.6, 6.5, 2.4 Hz, 1H), 2.80 - 2.52 (m, 2H), 2.36 (dt, 7= 13.6, 2.6 Hz, 1H), 2.162.05 (m, 1H), 1.84 (dd,7= 13.6, 11.8 Hz, 1H), 1.66 (dt, 7=13.4, 6.7 Hz, 1H), 1.47 (dd, 7= 13.6, 11.6 Hz, 1H), 0.95 (dd, 7= 9.1, 6.8 Hz, 6H). LCMS m/z 367.04 [M+H]⁺.
Compound 189 s Axr Ο H ί > \	0 Q H ( N S43 \	Compound 186⁴'⁵	'H NMR (300 MHz, Chloroform-7) δ 7.86 (s, 1H), 6.62 (s, 1H), 4.91 (dd, 7 = 12.7,3.0 Hz, 1H), 4.08 (s, 3H), 3.94 (q, 7=5.3 Hz, 2H), 3.63 (t, 7= 10.9 Hz, 1H), 2.87 - 2.50 (m, 4H), 2.36 (d, 7= 14.2 Hz, 1H), 2.17 (t, 7= 16.2 Hz, 3H), 2.09- 1.87 (m, 5H), 1.77 (d, 7= 9.0 Hz, 2H). LCMS m/z 379.06 [M+H]⁺.

258

Product	Piperidone	Method	¹H NMR; LCMS m/z [M+H]⁺
Compound 190 ^clACo H ί '.N A \	0 H ( N A S44 \	Compound 186 ³·⁸	Ή NMR (300 MHz, Chloroformé) δ 7.52 (s, 1H), 6.58 (s, 1H), 4.49 (dd,7 = 11.9, 2.7 Hz, 1H), 4.06 (s, 3H), 3.95 (t, 7=5.4 Hz, 2H), 3.60 - 3.38 (m,3H), 3.34 (s, 3H), 2.61 (q, 7=5.1 Hz, 2H), 2.38 (dd, 7= 13.7,2.8 Hz, 1H), 2.02-1.91 (m, 2H), 1.61 (dd, 7= 13.6, 11.7 Hz, 1H). LCMS m/z 369.07 [M+H]⁺.
Compound 191 «TÛ, \| H ί > A \	0 A^r'^JC\i^r^Ns' \| H £ n S42 \ [CIS]	Compound ₁₈₇3,6.7,8	*H NMR (300 MHz, Chloroformé) δ 7.78 (s, 1H), 7.14 (s, 1H), 4.75 (t, 7= 7.7 Hz, 1H), 4.08 (s, 3H), 3.97 (q, 7=5.1 Hz, 2H), 3.39 -3.20(m, 1H), 2.73 (q,7 = 5.7, 5.2 Hz, 2H), 2.42 (d, 7= 7.7 Hz, 2H), 2.23 (d,7= 14.2 Hz, 1H), 2.08 - 1.76 (m, 2H), 1.00 (dd, 7= 11.8, 6.8 Hz, 6H). LCMS m/z 401.11 [M+H]⁺.
Compound 192 ^fjcLu> ^S/^>A </ H £ > A \	0 Q H ί N ^-N S43 \	Compound 187 ^5,9	‘H NMR (300 MHz, Chloroformé) δ 7.91 (s, 1H), 7.16 (d, 7= 1.3 Hz, 1H), 4.97 (dd, 7= 12.8,3.0 Hz, 1H), 4.10 (s, 3H), 3.99 (hept, 7= 5.6 Hz, 2H), 3.79 3.53 (m, 1H), 2.92-2.68 (m, 3H), 2.61 (q, 7=8.1 Hz, 1H), 2.40 (d, 7= 14.5 Hz, 1H), 2.26 (d, 7=14.7 Hz, 1H), 2.02 (td, 7= 24.1, 21.4, 13.0 Hz, 6H), 1.80 (p,7= 9.3, 8.2 Hz, 2H). LCMS m/z 413.21 [M+H]⁺.

259

Product	Piperidone	Method	^!HNMR; LCMS m/z [M+H]⁺
Compound 193 f₃cA^z II 1 A n H ί 'N \	O H { N S44 \	Compound 187⁵·¹⁰	Ή NMR (300 MHz, Chloroform-7) δ 7.97 (s, 1H), 7.20 - 7.06 (m, 1H), 5.07 (dd, 7=12.7,3.2 Hz, 1H), 4.10 (s, 3H), 3.98 (h, 7= 6.1 Hz, 3H), 3.71 (dd, 7= 10.2, 3.3 Hz, 1 H), 3.57 (dd, 7= 10.2, 5.6 Hz, lH),3.39(s, 3H), 2.93 - 2.63 (m, 3H), 2.47 (d, 7= 14.9 Hz, 1H), 2.44 - 2.25 (m, 1H), 2.20 (s, 1H) LCMS m/z 403.04 [M+H]⁺.

1. The reaction stirred at 40 °C for 22 hours

2. Additional MsOH (31 pL, 4 equiv) was added and the reaction stirred at 40 °C for 4 hours then at room température ovemight

3. Purification by reversed-phase HPLC (Method: Cl8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H₂O with 0.2% formic acid) yielded the product

4. The reaction was stirred at 40 °C for 4 hours

5. Purification by reversed-phase HPLC (Method: C18 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H₂O with 0.1% trifluoroacetic acid) yielded the product

6. Additional MsOH (11 pL, 1.3 equiv) was added to the reaction and stirred at 80 °C for 24 hours

7. The reaction stirred at 40 °C for 2.5 hours

8. The reaction stirred at 65 °C for 4 days

9. The reaction stirred at 60 °C for 5 days .

Compound 194

2-chloro-2 6'-dimethyl-spiro[4,5-dihydrothieno[2,3-cjpyran-7,4 ’-piperidine] (194)

194

260

J

Step 1. Synthesis of2-chloro-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine] (194)

To a solution of 2,6-dimethylpiperidin-4-one C80 (Hydrochloride sait) (40 mg, 0.2444 mmol) and 2-(5-chloro-3-thienyl)ethanol S2 (39.75 mg, 30.23 pL, 0.2444 mmol) in DCM (1 mL) was added Triflic Acid (91.70 mg, 54.07 pL, 0.6110 mmol). The reaction was stirred at room température ovemight. The reaction was quenched with NaOH (2 M), and then extracted with DCM. Purification by reversed-phase HPLC (Method: Cl 8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 5 mM HCl) afforded 2-chloro-2',6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (Trifluoroacetate sait) 194. 'H NMR (300 MHz, Chloroform-7) δ 9.87 (s, 1H), 8.98 - 8.36 (m, 1H), 6.62 (s, 1H), 3.90 (t, J= 5.4 Hz, 2H), 3.61 (s, 2H), 2.64 (q, J= 5.4 Hz, 2H), 2.35 - 1.79 (m,4H), 1.35 (d, J = 6.6 Hz, 6H). LCMS m/z 272.03 [M+H]⁺.

Compounds 195 and 196

Compounds 195 and 196 (see Table 7) were prepared from a single Oxa-Pictet Spengler step with relevant piperidone and thiophene éthanol as described for compound 194. Thiophene ethanols were prepared by methods described above or obtained from commercial sources. Piperdinones were obtained from commercial sources. Any modifications to methods are noted in Table 7 and accompanying footnotes.

Table 7. Method of préparation, structure and physicochemical data for compounds 195-196

Product	Thiophene Ethanol And piperidone	Method	’HNMR; LCMS m/z [M+H]⁺
Compound 195 0 H	F S ^XOH S6 O H C81	Compound 194	‘H NMR (400 MHz, DMSO-A) δ 9.35 (s, 1H), 8.83 (s, 1H), 7.34 (s, 1H), 3.94 (hept, J= 5.9, 5.4 Hz, 2H), 3.75 - 3.64 (m, 1H), 3.57 (s, 1H), 2.87 (d, J= 5.6 Hz, 2H), 2.22 (dd, J =15.2,6.2 Hz, 1H), 2.12 (t, J= 13.3 Hz, 1H), 2.06-1.91 (m, 2H), 1.47 (d, 7=7.1 Hz, 3H), 1.29 (d, 7= 6.4 Hz, 3H). LCMS m/z. 306.24 [M+H]⁺

261

Product	Thiophene Ethanol And piperidone	Method	‘HNMR; LCMS m/z [M+H[
Compound 196 oCÏM^f H	F S OH ^Γ S6 O C82 H	Compound 194	LCMS m/z 306.24 [M+H]⁺

Préparation of S45 ~[(2R, 6S)-2 ' 6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 '-piperidine]-l '-yl]-2,2,2trifluoro-ethanone (S45)

Step 1. Synthesis of (2R,6S)-2',6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine] (C83)

To a solution of (2S,6R)-2,6-dimethylpiperidin-4-one C82 (900 mg, 7.076 mmol) in dioxane (21 mL) at 0 °C was added 2-(3-thienyl)ethanol SI (850 mg, 6.631 mmol) followed by Triflic Acid (2 g, 13.33 mmol). The reaction was warmed to room température. After stirring for 2 hours the reaction had gone to completion. The reaction was carefully quenched with saturated sodium bicarbonate solution. The reaction mixture was partitioned between DCM and aqueous saturated sodium bicarbonate solution. The organic phase was separated, passed through a phase separator, and concentrated via rotary évaporation to give crude (2R,6S)-2',6'-dimethylspiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] C83 (1.059 g, quant.). LCMS m/z 238.11 [M+H]⁺.

Step 2. Synthesis of l-[(2R,6S)-2',6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-1 ’-yl]-2,2,2-trifluoro-ethanone (S45) (2R,6S)-2',6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] C83 was dissolved in DCM (12.7 mL) and BociO (1.4 g, 6.415 mmol) and DIPEA (1.7 g, 13.15 mmol) were added. The reaction was stirred for five hours but only small amounts of conversion were observed. The reaction mixture was concentrated to dryness and re-dissolved in DCM (12.7 mL). To this solution at 0 °C was added TEA (1.34 g, 13.24 mmol) and TF AA (1.8 g, 8.570 mmol). The reaction was warmed to room température and stirred for two hours until the reaction had 262

J gone to completion. The reaction was quenched with saturated sodium bicarbonate and extracted with EtOAc (2x). The organics were washed with brine, dried over Na₂SÛ4, and concentrated in vacuo. Purification by silica gel chromatography (Column: 24g silica gel, Gradient: 0-50% EtOAc in Heptane) afforded l-[(2R,6S)-2',6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-r-yl]-2,2,2-trifluoro-ethanone S45 (1 g, 67%). *H NMR (300 MHz, Chloroform-7) δ 7.19 (d, J= 5.0 Hz, 1H), 6.78 (d, J= 5.0 Hz, 1H), 4.47 (d, 7= 7.8 Hz, 2H), 3.85 (t, J= 5.5 Hz, 2H), 2.68 (t, 7= 5.5 Hz, 2H), 2.50 (dd, 7= 14.4, 8.2 Hz, 2H), 1.97 (dd, 7= 14.7, 6.6 Hz, 2H), 1.49 (dd, 7= 20.4, 6.6 Hz, 6H). LCMS m/z 334.05 [M+H]⁺.

400 mg of the boc protected compound was also isolated during purification. 'H NMR (300 MHz, Chloroform-7) δ 7.18 (dd, 7= 7.6, 5.0 Hz, 1H), 6.76 (d, 7= 5.0 Hz, 1H), 4.36 (h, 7= 7.1 Hz, 2H), 3.88 (t, 7= 5.5 Hz, 2H), 2.68 (t, 7= 5.5 Hz, 2H), 2.49 - 2.31 (m, 2H), 1.83 (dd, 7 = 14.5,7.0 Hz, 2H), 1.51 (d, 7= 25.0 Hz, 9H), 1.33 (d, 7= 6.8 Hz, 6H).

Compound 197 (2R, 6S)-2-bromo-26'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4 -piperidine](197)

Step 1. Synthesis of l-[(2R,6S)-2-bromo-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3c]pyran- 7,4 '-piperidine]-1 ’-yl]-2,2,2-trifluoro-ethanone (S46)

To a solution of l-[(2R,6S)-2',6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-r-yl]-2,2,2-trifluoro-ethanone S45 (1 g, 3.000 mmol) dissolved in MeCN (10.7 mL) was added NBS (610 mg, 3.427 mmol) and the reaction was stirred at 65 °C for one hour. The solution was concentrated and dissolved in DCM. The reaction was quenched with aqueous sodium thiosulfate and DCM. The organic phase was separated, passed through a phase separator, and concentrated in vacuo and the solid NBS was fïltered off. Purification by silica gel chromatography (Gradient: 0-60% EtOAc in heptane) yielded the product l-[(2R,6S)-2-bromo2',6^,-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-l'-yl]-2,2,2-trifluoroethanone S46 (1.12 g, 76%). Ή NMR (300 MHz, Chloroform-7) δ 6.74 (s, 1H), 4.61 - 4.32 (m,

263

2H), 3.83 (t, J= 5.5 Hz, 2H), 2.65 - 2.40 (m, 4H), 1.88 (dd, J= 14.9, 6.5 Hz, 2H), 1.43 (d, J = 6.9 Hz, 6H). LCMS m/z 412.05 [M+H]⁺.

Step 2. Synthesis of (2R,6S)-2-bromo-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran7,4'-piperidine] (197)

To a solution of l-[(2R,6S)-2-bromo-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran7,4'-piperidine]-T-yl]-2,2,2-trifluoro-ethanone S46 (1.12 g, 2.716 mmol) in DCM (10 mL) was added aqueous NaOH (1.5 mL of 2 M, 3.000 mmol). The reaction was heated to 40 °C. The reaction was stirred for 30 minutes. The organic phase was separated, passed through a phase separator, and concentrated in vacuo. Purification by reversed-phase HP LC (Method: Cl 8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 0.1% trifluoroacetic acid) yielded (2R,6S)-2-bromo-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] (trifluoroacetate sait) 197 (11.9 mg, 1%). *H NMR (300 MHz, Chloroformé) δ 9.43 (s, 1H), 8.79 - 8.07 (m, 1H), 6.76 (s, 1H), 3.90 (t, J= 5.4 Hz, 2H), 3.64 (d, J= 8.0 Hz, 2H), 2.66 (t, J= 5.4 Hz, 2H), 2.17 (d, J= 14.4 Hz, 2H), 1.93 (dd, J= 14.5, 12.2 Hz, 2H), 1.35 (d, J= 6.6 Hz, 6H). LCMS m/z 316.18 [M+H]⁺.

Compound 198 (2R,6S)-2-(3,3-difluorocyclobutyl)-2^,,6^,-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine] (198)

Step 1. Synthesis of l-[(2R,6S)-2- (3,3-difluorocyclobutyl)-2',6'-dimethyl-spiro[4,5dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-1 ’-yl]-2,2,2-trifluoro-ethanone (C84)

To Ir[df(CF3)ppy]2(dtbbpy)PFô (Phosphorus Hexafhioride Ion) (3 mg, 0.002971 mmol), Dichloro(dimethoxyethane)nickel (4 mg, 0.01820 mmol), and 4-tert-butyl-2-(4-tert-butyl-2pyridyl)pyridine (5 mg, 0.01863 mmol) under an inert atmosphère was added a solution of 1[(2R,6S)-2-bromo-2',6^,-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]2,2,2-trifluoro-ethanone S46 (65 mg, 0.1577 mmol), bis(trimethylsilyl)silyl-trimethyl-silane (61 pL, 0.1963 mmol), and 2,6-dimethylpyridine (41.64 mg, 45.01 pL, 0.3886 mmol) dissolved in DME (2 mL). 3-bromo-l,l-difluoro-cyclobutane (158 pL, 1.571 mmol) was added. Theresulting

264 mixture was sealed and irradiated in a Sigma SynLED photoreactor ovemight with tumble stirring. The reaction vial was unsealed and evaporated under a stream of nitrogen. The resulting residues were diluted with 2 mL water and 2 mL DCM and stirred for several minutes. The biphasic mixtures were passed through a parallel hydrophobie filter plate. The DCM layers were concentrated in vacuo. Purification by reversed-phase HP LC (Method: Cl 8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 0.1% trifluoroacetic acid) yielded TFA protected intermediate C84. LCMS m/z 424.18 [M+H]⁺.

Step 2. Synthesis of (2R,6S)-2-(3,3-difluorocyclobutyl)-2',6'-dimethyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4 ’-piperidine] (198)

To l-[(2R,6S)-2- (3,3-difluorocyclobutyl)-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone (15.5 mg, 0.037 mmol) C84 in DCM (2 mL) was added NaOH 6M (10 eq). The reaction was heated to 40 °C and stirred for 5 hours. The organic phase was separated, passed through a phase separator, and concentrated in vacuo. Purification by reversed-phase HP LC (Method: Cl 8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 0.1% trifluoroacetic acid) yielded deprotected product (2R,6S)-2-(3,3-difluorocyclobutyl)-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine] (Trifluoroacetate sait) 198 (5.6 mg, 8%). *H NMR (300 MHz, Chloroform-J) δ 6.50 (d, J= 1.0 Hz, 1H), 3.84 (t, J= 5.4 Hz, 2H), 3.73 - 3.28 (m, 3H), 2.97 (tdd, J= 14.3, 7.3, 4.1 Hz, 2H), 2.79 - 2.35 (m, 5H), 2.10 (d, J= 13.8 Hz, 2H), 2.02 - 1.74 (m, 2H), 1.28 (d, J= 6.6 Hz, 7H).

Compounds 199-202

Compounds 199-202 (see Table 8) were prepared from intermediate S47 using the appropriate reagent and using the photo-redox method and deprotection method as described for compound 198. Alkyl bromides were obtained from commercial sources. Any modifications to methods are noted in Table 8.

Table 8. Method of préparation, structure and physicochemical data for compounds 199-202

Product	Alkyl Bromide	Method	Ή NMR; LCMS m/z [M+IIf
Compound 199 H	Xx^Br	Compound 198	*H NMR (300 MHz, Chloroformé δ 9.55 (s, 1H), 8.55 (s, 1H), 6.48 (d, J= 1.1 Hz, 1H), 3.90 (t, J= 5.4 Hz, 2H), 3.65 (s, 2H), 2.93 - 2.45 (m, 4H), 2.27 - 1.76 (m, 4H), 1.49 1.17 (m, 9H). LCMS m/z 266.28 [M+H]⁺.

265

Product	Alkyl Bromide	Method	'H8MR; LCMS m/z [M+Hf
Compound 200 oJU \ H	^Br	Compound 198	‘H NMR (300 MHz, Chloroform-iZ) δ 9.51 (s, 1H), 8.50 (s, 1H), 6.49 (d, J= 0.9 Hz, 1H), 3.90 (t, J= 5.4 Hz, 2H), 3.64 (s, 2H), 3.12 (hept, J = 6.9 Hz, 1H), 2.64 (t, J= 5.4 Hz, 2H), 2.27 - 1.82 (m, 4H), 1.34 (dd, J = 9.0, 6.7 Hz, 12H). LCMS m/z 280.32 [M+H]⁺.
Compound 201 H	J^Br	Compound 198	‘H NMR (300 MHz, Chloroform-cf) δ 6.44 (s, 1H), 3.90 (t, J= 5.4 Hz, 2H), 3.84 - 3.49 (m, 2H), 2.83 2.47 (m, 5H), 2.18 (d, J= 14.5 Hz, 2H), 2.02 - 1.78 (m, 3H), 1.35 (d, J = 6.6 Hz, 6H), 0.96 (d, J= 6.6 Hz, 6H). LCMS m/z 294.32 [M+H]⁺.
Compound 202 H		Compound 198	‘H NMR (300 MHz, Chloroformé) δ 9.60 (s, 1H), 8.57 (d, J= 12.5 Hz, 1H), 6.47 (s, 1H), 3.64 (s, 2H), 3.35 (s, 2H), 2.74 (s, 1H), 2.64 (t, J= 5.3 Hz, 2H), 2.26 - 1.68 (m, 9H), 1.61 1.04 (m, 11H). LCMS m/z 320.5 [M+H]+.

Compound 203 (2R, 68)-2-(2,2-difluoroethyl)-2 ', 6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4'piperidine] (203)

Step 1. Synthesis of tert-butyl (2R,6S)-2',6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran7,4'-piperidine]-1 ’-carboxylate (C85)

266

To a solution of (2R,6S)-2',6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine] C83 in DCM (4 mL) was added BOC2O (525 pL, 2.285 mmol) and DIPEA (597 pL, 3.427 mmol). The reaction was stirred ovemight until the reaction had gone to completion. The reaction was quenched with saturated sodium bicarbonate and extracted with EtOAc (2x). The organics were washed with brine, dried over anhydrous Na2SO₄, and concentrated in vacuo. Purification by silica gel chromatography (Column: 24 g silica gel, Gradient: 0-30% EtOAc in Heptane) afforded teri-butyl (2R,6S)-2',6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-r-carboxylate C85 (210 mg, 54%). LCMS m/z 338.1 [M+H]⁺;

Step 2. Synthesis of tert-butyl (2R,6S)-2-bromo-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-1 '-carboxylate (C86)

To a solution of ierZ-butyl (2R,6S)-2',6'-dimethylspiro[4,5-dihydrothierio[2,3-c]pyran7,4'-piperidine]-r-carboxylate C85 (140 mg, 0.4148 mmol) in MeCN (1.5 mL) was added NBS (84 mg, 0.4720 mmol). The reaction was stirred at 65 °C for one hour. The reaction was quenched with a solution of sodium thiosulfate and DCM. The organic phase was separated, passed through a phase separator, and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-30% EtOAc in heptane) yielded the product tert-butyl (2R,6S)-2bromo-2^,,6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-carboxylate C86 (120 mg, 69%). LCMS m/z 416.14 [M+H]⁺.

Step 3. Synthesis of tert-butyl (2R,6S)-2-(2,2-difluoroethyl)-2',6'-dimethyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4 '-piperidine]-1 '-carboxylate (C87)

To vial was added Ir[df(CF3)ppy]2(dtbbpy)PF6 (Phosphorus Hexafluoride Ion) (3 mg, 0.002971 mmol), Dichloro(dimethoxyethane)nickel (4 mg, 0.01820 mmol), and 4-tert-butyl-2(4-tert-butyl-2-pyridyl)pyridine (5 mg, 0.01863 mmol) as solids. The vial was purged and refïlled with N2 (3x). To the vial was added sequentially tert-butyl (2R,6S)-2-bromo-2',6'dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-carboxylate C86 (70 mg, 0.1680 mmol), bis(trimethylsilyl)silyl-trimethyl-silane (51 mg, 0.2051 mmol), 2,6dimethylpyridine (45 mg, 0.4200 mmol) and DME (2 mL). Purged with N2 (3x) and 2-bromo1,1-difluoro-ethane (244 mg, 1.683 mmol) was added. The reaction was irradiated with Merck Integrated Photoreactor for 7 h, Royal Blue (450 nm) LED light. 100% LED light power was applied. The stir rate was 1000 rpm. The reaction was quenched with aqueous saturated sodium bicarbonate and DCM and the organic layer was collected through a phase separator. The solvent was concentrated in vacuo to give crude ZerZ-butyl-(2R,6S)-2-(2,2-difluoroethyl)-2',6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-carboxylate C87 (8 mg, 12%). LCMS m/z 402.23 [M+H]⁺.

267

Step 4. Synthesis of (2R,6S)-2-(2,2-difluoroethyl)-2',6'-dimethyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] (203)

To a solution of ZerZ-butyl-(2R,6S)-2-(2,2-difluoroethyl)-2',6'-dimethyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-carboxylate C87 (8 mg, 0.02 mmol) in Dioxane (350 pL) was added HCl (630 pL of 4 M, 2.520 mmol) in dioxane. The reaction was stirred for 3 hours. Solvent was removed in vacuo. Purification by reversed-phase HP LC (Method: Cl 8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H₂O with 0.1% trifluoroacetic acid) yielded (2R,6S)-2-(2,2-difluoroethyl)-2',6'-dimethyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] (Trifluoroacetate sait) 203 (6.1 mg, 78%). 'H NMR (300 MHz, Chloroform-7) δ 9.81 (s, 1H), 8.80 (s, 1H), 6.65 (s, 1H), 5.93 (tt, J= 56.4, 4.4 Hz, 1H), 3.90 (t, 7= 5.4 Hz, 2H), 3.30 (td, 7= 16.7, 4.5 Hz, 3H), 2.67 (t, 7= 5.5 Hz, 2H), 2.30 - 1.75 (m, 4H), 1.36 (d, J= 6.5 Hz, 6H). LCMS m/z 302.22 [M+H]⁺.

Compounds 204 and 205 (2R,6S)-2-chloro-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (204) and (2R, 6S)-2-chloro-2 6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-4-ol (205)

Step 1. Synthesis of (2R,6S)-2-chloro-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran7,4'-piperidine] (204)

To a solution of (2S,6R)-2,6-dimethylpiperidin-4-one (Hydrochloride sait) C82 (98 mg, 0.5989 mmol) and 2-(5-chloro-3-thienyl)ethanol S2 (100 pL, 0.8184 mmol) in DCM (2 mL) was added MsOH (200 pL, 3.082 mmol) and the mixture was stirred at 40 °C for 1 hour. The mixture was cooled to room température, and the pH was adjusted to pH 14 with aqueous NaOH. The

268 aqueous layer was extracted with additional DCM (2 mL). The combined organic layer was passed through a phase separator and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-10% MeOH in DCM) yielded the product which was diluted in diethyl ether (4 mL). HCl (200 pL of 4 M, 0.8000 mmol) was added to sait the product, and the solid was filtered, rinsed with additional ether and dried to yield (2R,6S)-2-chloro-2',6'-dimethylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4^,-piperidine] (Hydrochloride sait) 204 (102 mg, 55%). *H NMR (400 MHz, DMSO-J₆) δ 9.21 (d, J= 10.4 Hz, 1H), 8.82 (d, J= 10.2 Hz, 1H), 6.93 (s, 1H), 3.89 (t, J= 5.4 Hz, 2H), 3.36 (d, J= 8.5 Hz, 2H), 2.58 (t, J= 5.4 Hz, 2H), 2.17 (d, J= 14.1 Hz, 2H), 1.82 (dd, J= 14.3, 12.2 Hz, 2H), 1.27 (d, J= 6.5 Hz, 6H). LCMS m/z 272.09 [M+H]⁺.

Step 2. Synthesis of l-[(2R,6S)-2-chloro-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3c]pyran- 7,4 ’-piperidine]-1 ’-yl] -2,2,2-trifluoro-ethanone (C88)

To a solution of (2R,6S)-2-chloro-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran7,4'-piperidine] (Hydrochloride sait) 204 (319 mg, 1.173 mmol) in DCM (6 mL) was added TF AA (750 mg, 3.571 mmol) and TEA (600 mg, 5.929 mmol). The reaction was stirred for 30 minutes at room température. The reaction was quenched with sodium bicarbonate and DCM. The organic phase was separated, passed through a phase separator, and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-50% EtOAc in heptane) yielded the product l-[(2R,6S)-2-chloro-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]l'-yl]-2,2,2-trifluoro-ethanone C88 (388 mg, 85%). LCMS m/z 368.08 [M+H]⁺.

Step 3. Synthesis of (2R,6S)-2'-chloro-2,6-dimethyl-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4 '-one (C89)

To a solution of l-[(2R,6S)-2-chloro-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C88 in MeCN (10 mL) was added cobaltous acetate tetrahydrate (128 mg, 0.5139 mmol) and N-hydroxyphthalimide (168 mg, 1.030 mmol). The reaction was purged and evacuated with oxygen (3x) and then heated to 45 °C under an oxygen balloon. The reaction was stirred for 7 hours. The reaction was diluted with water and DCM. The organic phase was separated, passed through a phase separator, and concentrated in vacuo. The material was brought up in a minimal amount of DCM and a solid began crashing out of solution. The liquid was decanted off. Purification by silica gel chromatography (Gradient: 060% EtOAc in Heptane) afforded (2R,6S)-2'-chloro-2,6-dimethyl-1-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one C89 (122 mg, 31%). *H NMR (300 MHz, Chloroform-7) δ 7.21 (s, 1H), 4.63 - 4.39 (m, 2H), 4.29 (s, 2H), 2.69 (dd, J= 14.4, 8.1 Hz, 2H), 2.07 - 1.80 (m, 2H), 1.48 (d, J= 6.9 Hz, 6H). LCMS m/z 382.03 [M+H]⁺.

269

Step 4. Synthesis of l-[(2R,6S)-2-chloro-4-hydroxy-2',6'-dimethyl-spiro]4,5dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-1 ’-yl]-2,2,2-trifluoro-ethanone (C90)

To a solution of (2R,6S)-2'-chloro-2,6-dimethyl-l-(2,2,2-trifluoroacetyl)spiro[piperidine4,7'-thieno[2,3-c]pyran]-4'-one C89 (122 mg, 0.3195 mmol) in DCM (2.44 mL) and MeOH (620 pL) was added NaBLL (73 mg, 1.930 mmol). The reaction was stirred for 45 minutes. The reaction was quenched with aqueous HCl and DCM. The organic phase was separated, passed through a phase separator, and concentrated in vacuo to give l-[(2R,6S)-2-chloro-4-hydroxy2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoroethanone C90 (122 mg, 98%). LCMS m/z 384.28 [M+H]⁺.

Step 5. Synthesis of (2R,6S)-2-chloro-2',6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran7,4'-piperidine]-4-ol (205)

To a solution ofl-[(2R,6S)-2-chloro-4-hydroxy-2',6'-dimethyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C90 (122 mg, 0.3195 mmol) in MeOH (620 pL) was added NaOH 6M (10 eq). The reaction was heated to 40 °C and stirred for 5 hours. The reaction was quenched with water and DCM. The organic layer was separated through a phase separator and concentrated in vacuo to give pure (2R,6S)-2-chloro2^,,6'-dimethyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol 205 (55 mg, 57%). ’H NMR (300 MHz, Methanol-7/). δ 6.85 (s, 1H), 4.43 (t, 7= 3.9 Hz, 1H), 3.96 (dd, 7= 12.2, 3.7 Hz, 1H), 3.73 (dd, 7= 12.1, 4.2 Hz, 1H), 3.22 - 2.92 (m, 2H), 2.14 - 1.96 (m, 2H), 1.38 -1.16 (m, 3H), 1.08 (dd, 7= 6.5, 4.1 Hz, 6H). LCMS m/z 288.06 [M+H]⁺.

Compound 206 (2R, 6S)-2-chloro-2 6'-dicyclopropyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4’-piperidine] (206)

Step 1. Synthesis of (2R,6S)-2-chloro-2',6'-dicyclopropyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] (206)

To a solution of 2,6-dicyclopropylpiperidin-4-one C91 (30 mg, 0.1674 mmol) in DCM (1000 pL) was added 2-(5-chloro-3-thienyl)ethanol S2 (25 pL) followed by MsOH (44 pL, 0.6780 mmol) and the mixture was stirred at reflux. After 5 minutes, the mixture was cooled, pH

270 adjusted with aqueous NaOH (150 pL of 6 M, 0.9000 mmol) to pH 14 and the organic layer was separated and concentrated. Silica gel purification (Gradient: 0-20% MeOH in DCM) yielded a mixture of cis and trans isomers. SFC purification yielded (2R,6S)-2-chloro-2',6'-dicyclopropylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] 206 (5.6 mg, 10%). *H NMR (300 MHz, Methanol-J#) δ 6.66 (s, 1H), 3.84 (t, J= 5.5 Hz, 2H), 2.57 (t, J= 5.5 Hz, 2H), 2.24 - 2.11 (m, 2H), 2.06 (ddd, 7 = 11.5, 9.0, 2.4 Hz, 2H), 1.54 (dd, 7= 13.3, 11.5 Hz, 2H), 0.87 - 0.69 (m, 2H), 0.62 - 0.39 (m, 4H), 0.37 - 0.21 (m, 2H), 0.21 - 0.04 (m, 2H). LCMS m/z 324.02 [M+H]+.

Compound 207 (2R, 6S)-2 ', 6'-dimethyl-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 'piperidine] (207)

Step 1. Synthesis of (2R,6S)-2',6'-dimethyl-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] (207)

To a solution of tert-butyl (2S,6R)-2,6-dimethyl-4-oxo-piperidine-l-carboxylate C92 (300 mg, 1.320 mmol) and 2-[5-(trifluoromethyl)-3-thienyl]ethanol S3 (300 mg, 1.483 mmol) in dioxane (4.5 mL) cooled to 0 °C was added Triflic Acid (345 pL, 3.899 mmol). The reaction was warmed to room température. After 10 minutes ketal formation was observed. The solution was stirred for 6 hours. The solution was diluted with DCM and washed with 2 M NaiCCh. The reaction was extracted with DCM (3x), dried with Na2SO4, filtered, and the DCM was removed in vacuo to give crude (2R,6S)-2^,,6'-dimethyl-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] 207 (400 mg, 71%). LCMS m/z 306.06 [M+H]⁺.

271

Step 2. Synthesis of tert-butyl (2R,6S)-2',6'-dimethyl-2-(trifluoromethyl)spiro[4,5dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-1 '-carboxylate (C93)

To a solution of (2R,6S)-2',6'-dimethyl-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] 207 (400 mg, 71%) in DCM (8 mL) was added boc anhydride (1.5 mL, 6.529 mmol) and DIPEA (670 pL, 3.847 mmol). The reaction was stirred for four days until full conversion was observed. The solvent was concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-30% EtOAc in heptane) yielded the product tert-butyl (2R,6S)2',6'-dimethyl-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-rcarboxylate C93 (440 mg, 82%). *HNMR (300 MHz, Chloroformé δ 7.11 (d, J= 1.3 Hz, 1H), 4.47 - 4.25 (m, 2H), 3.89 (t, J= 5.5 Hz, 2H), 2.67 (t, J= 5.5 Hz, 2H), 2.43 (ddd, J= 15.5, 8.5, 2.3 Hz, 2H), 1.79 (dd, J= 14.5, 6.9 Hz, 2H), 1.50 (s, 9H), 1.33 (d, J= 6.8 Hz, 6H).

Step 3. Synthesis of (2R,6S)-2’,6'-dimethyl-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] (207)

To a solution of tert-butyl (2R,6S)-2',6'-dimethyl-2-(trifluoromethyl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-carboxylate C93 (20 mg, 0.049 mmol) dissolved in dioxane (100 pL) was added HCl (100 pL of 4 M, 0.4000 mmol) in dioxane. The reaction was stirred at room température for 1 hour. The solvent was concentrated in vacuo. Purification by reversed-phase HPLC (Method: Cl8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 0.1% trifluoroacetic acid) afforded (2R,6S)-2',6'-dimethyl-2(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (Trifluoroacetate sait) 207 (6.6 mg, 32%). ‘H NMR (300 MHz, Chloroformé δ 9.55 (s, 1H), 8.67 (s, 1H), 7.16 (d, J= 1.3 Hz, 1H), 3.94 (t, J= 5.4 Hz, 2H), 3.67 (s, 2H), 2.74 (t, J= 5.4 Hz, 2H), 2.38 - 1.84 (m, 4H), 1.37 (d, J= 6.6 Hz, 6H). LCMS m/z 306.24 [M+H]⁺.

272

Compound 208

[(2R, 6S)-2 6'-dimethyl-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 'piperidine]-3-yl]methanol (208)

Step 1. Synthesis of tert-butyl (2R,6S)-3-formyl-2',6'-dimethyl-2-(trifluoromethyl)spiro[4,5dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-1 '-carboxylate (C94)

To a solution of tert-butyl (2R,6S)-2',6'-dimethyl-2-(trifluoromethyl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-carboxylate C93 (420 mg, 1.036 mmol) in THF (6 mL) at -78 °C under N2 was added s-BuLi (1 mL of 1.4 Μ, 1.400 mmol). The reaction was stirred for 30 minutes, followed by addition of DMF (160 pL, 2.066 mmol). The reaction was stirred for 30 minutes and warmed to room température. The reaction was quenched with NH4CI and DCM. The organic phase was separated, passed through a phase separator, and concentrated to give crude tert-butyl (2R,6S)-3-formyl-2',6'-dimethyl-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-r-carboxylate C94. LCMS m/z 434.32 [M+H]⁺.

Step 2. Synthesis of tert-butyl (2R,6S)-3-(hydroxymethyl)-2',6'-dimethyl-2(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-1 '-carboxylate (C95) The reaction was stirred for 30 minutes. The reaction was quenched with aqueous sodium bicarbonate and DCM. The organic phase was separated, passed through a phase separator, and concentrated in vacuo to give crude tert-butyl (2R,6S)-3-(hydroxymethyl)-2',6'-dimethyl-2(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-carboxylate C95 LCMS m/z 436.35 [M+H]⁺.

273

Step 3. Synthesis of (2R,6S)-2\6'-dimethyl-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-3-yl]methanol (208)

To a solution of crude C95 in dioxane (2 mL) was added HCl (2.5 mL of 4 M, 10.00 mmol) in dioxane. The reaction was stirred for 1 hour. Solvent was removed in vacuo.

Purification by reversed-phase HPLC (Method: Cl8 Waters Sunfire column (30 x 150 mm, 5 micron)) afforded (2'S ,6^,R)-2',6'-dimethyl-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-3-yl]methanol 208 (192 mg, 55%). 'H NMR (300 MHz, Chloroform-7) δ 4.66 (d, J= 1.4 Hz, 2H), 3.98 (t, J= 5.5 Hz, 2H), 3.17 (dtt, J= 12.6, 6.3, 3.1 Hz, 2H), 2.75 (t, J = 5.5 Hz, 2H), 2.19-1.97 (m, 2H), 1.35 (dd, J= 13.5, 11.4 Hz, 2H), 1.10 (d, 7=6.4 Hz, 6H).

LCMS m/z 336.08 [M+H]+.

Compound 209 methyl (2 'S, 6'R, 7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-2 carboxylate (209)

274

Step 1. Synthesis of dimethyl (2S)-2-(tritylamino)butanedioate (C97)

To a mixture of dimethyl (2S)-2-aminobutanedioate C96 (Hydrochloride sait) (2000 mg, 10.12 mmol) and DIPEA (3.7 mL, 21.24 mmol) in DCM (40 mL) was added [chloro(diphenyl)methyl]benzene (3 g, 10.76 mmol). The mixture was stirred at room température ovemight. The mixture was quenched with water, phase separated, and the organic layer was dried with MgSCU, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-30% EtOAc in heptane) yielded dimethyl (2S)-2(tritylamino)butanedioate (3.11 g, 75%) C97. *H NMR (300 MHz, Chloroform-7) δ 7.59 - 7.43 (m, 6H), 7.35 - 7.24 (m, 6H), 7.24 - 7.13 (m, 3H), 3.70 (s, 4H), 3.28 (s, 3H), 2.95 (d, J= 10.1 Hz, 1H), 2.67 (dd, J= 14.7, 5.4 Hz, 1H), 2.53 (dd, J= 14.7, 7.0 Hz, 1H). LCMS m/z 402.23 [M+H]⁺.

Step 2. Synthesis of methyl (2S)-5-dimethoxyphosphoryl-4-oxo-2-(tritylamino)pentanoate (C98)

To a mixture of [methoxy(methyl)phosphoryl]oxymethane (2 mL, 18.46 mmol) in THF (90 mL) cooled to -78 °C was added a solution of sec-butyllithium (13.5 mL of 1.4 Μ, 18.90 mmol), dropwise. After 5 minutes at this température, a solution of dimethyl (2S)-2(tritylamino)butanedioate C97 (3100 mg, 7.683 mmol) in THF (10 mL) was added, dropwise, and the mixture was stirred for 10 additional minutes and then quenched with AcOH (1.3 mL, 22.86 mmol). The mixture was diluted with water (100 mL) and ether (100 mL). The organic layer was removed, and subsequently washed with water (100 mL), brine (100 mL), dried with MgSCU, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-20% MeOH in DCM) yielded methyl (2S)-5-dimethoxyphosphoryl-4-oxo-2-(tritylamino)pentanoate C98 (1580 mg, 41%). Ή NMR (300 MHz, Chloroform-J) δ 7.56 - 7.42 (m, 6H), 7.30 (t, J= 1.6 Hz, 3H), 7.28 - 7.16 (m, 6H), 3.81 (s, 3H), 3.78 (s, 3H), 3.72 (s, 1H), 3.31 (s, 3H), 3.07 (d, J= 22.6 Hz, 2H), 2.91 (dd, J= 16.7, 4.6 Hz, 2H), 2.79 (dd, J= 16.7, 6.9 Hz, 1H). LCMS m/z 494.31 [M+H]⁺.

Step 3. Synthesis of methyl (E,2S)-4-oxo-2-(tritylamino)hept-5-enoate (C99)

To a mixture of methyl (2S)-5-dimethoxyphosphoryl-4-oxo-2-(tritylamino)pentanoate C98 (553 mg, 1.108 mmol) in MeCN (30 mL) was added potassium carbonate (170 mg, 1.230 mmol) followed by acetaldehyde (200 pL, 3.564 mmol). The mixture was warmed to 50 °C and stirred ovemight. The mixture was cooled to room température, concentrated, and re-diluted in ethyl acetate (50 mL) and water (30 mL) and brine (10 mL). The organic layer was separated, washed with brine (30 mL), dried with MgSÛ4, filtered, and concentrated. Purification by silica

275 gel chromatography (Gradient: 0-40% EtOAc in Heptane) yielded methyl (E,2S)-4-oxo-2(tritylamino)hept-5-enoate C99 (282 mg, 61%). LCMS m/z 412.27 [M+H]⁺.

Step 4. Synthesis of methyl (2S,6S)-6-methyl-4-oxo-piperidine-2-carboxylate (C100) and methyl (2S,6R)-6-methyl-4-oxo-piperidine-2-carboxylate (C101)

To a mixture of methyl (E,2S)-4-oxo-2-(tritylamino)hept-5-enoate C99 (187 mg, 0.4203 mmol) in methanol (30 mL) at room température was added hydrochloric acid (7500 pL of 2 M, 15.00 mmol). The mixture was stirred for 10 minutes. Intermediate, methyl (E,2S)-2-amino-4oxo-hept-5-enoate, was formed. To the mixture was added water (15 mL) and DIPEA (4500 pL, 25.84 mmol) and the reaction was stirred for 15 minutes. The mixture was diluted with EtOAc (60 mL), water (60 mL), and the layers were separated. The aqueous layer was extracted with EtOAc (2 x 50 mL), and the combined organic layer was dried with MgSO4, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-10% MeOH in DCM) yielded C100 and ClOlas a 2:1 mixture of cis:trans isomers (28 mg, 39%).

C100 [DIAST-1] Ή NMR (300 MHz, Chloroformé) δ 3.75 (s, 3H), 3.64 (dd, J= 12.2, 3.5 Hz, 1H), 2.97 (dqd, J= 12.3, 6.2, 2.9 Hz, 1H), 2.72 - 2.53 (m, 2H), 2.43 - 2.34 (m, 1H), 2.08 (ddd, J= 14.2, 11.7, 1.0 Hz, 1H), 1.24 (d, J= 6.2 Hz, 3H). LCMS m/z 172.0 [M+H]⁺.

C101 [DIAST-2] *H NMR (300 MHz, Chloroformé) δ 4.06 (dd, J= 6.6, 3.7 Hz, 1H), 3.73 (s, 3H)), 3.32 - 3.15 (m, 1H), 2.72 - 2.53 (m, 2H), 2.43 - 2.34 (m, 1H), 2.08 (ddd, J= 14.2, 11.7, 1.0 Hz, 1H), 1.17 (d, 7= 6.2 Hz, 3H). LCMS m/z 172.0 [M+H]⁺.

Step 5. Synthesis of methyl (2'S,6'S,7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3c]pyran- 7,4 ’-piperidine]-2 '-carboxylate (209)

To a mixture C100 and C101(28 mg, 0.1636 mmol) (2:1 cis to trans) in DCM (1000 pL) was added 2-(5-chloro-3-thienyl)ethanol S2 (25 pL, 0.2021 mmol) followed by MsOH (50 pL, 0.7705 mmol) and the mixture was refluxed for 5 minutes. The mixture was cooled, and the pH was adjusted with aqueous NaOH (150 pL of 6 M, 0.90 mmol) to a pH of 14 and the organic layer was separated and concentrated. Purification by silica gel chromatography (Gradient: 020% MeOH in DCM) yielded methyl (2'S,6'S,7S)-2-chloro-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-2'-carboxylate 209 (24 mg, 68%). ’H NMR (300 MHz, Chloroformé) δ 6.57 (s, 1H), 3.92 (t, J= 5.5 Hz, 2H), 3.84 (dd, 7= 11.9, 2.8 Hz, 1H), 3.71 (s, 3H), 3.12 (dtd, 7= 12.7, 6.4, 2.6 Hz, 1H), 2.69 - 2.51 (m, 2H), 2.40 (dt, 7= 13.7, 2.7 Hz, 1H), 1.98 (dt, 7= 13.7, 2.6 Hz, 1H), 1.62 (dd,7= 13.6, 11.9 Hz, 1H), 1.33 (dd,7= 13.7, 11.3 Hz, 1H), 1.12 (d, 7= 6.3 Hz, 3H). LCMS m/z 316.01 [M+H]⁺. Minor 2,6-trans isomer was purged during isolation.

276

Compound 210 (2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-2'-yl)methanol (210)

C105

210

Step 1. Synthesis of [(2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-2'-yl]methyl benzoate (CIO5)

To a solution of 4-aminopentan-2-one C102 (50 mg, 0.36 mmol) in EtOH (1.5 mL) was added ΕίβΝ (51 pL, 0.3659 mmol) followed by 2- 2-oxoethyl benzoate C103 (65 mg, 0.396 mmol), L-proline (9 mg, 0.07817 mmol), and MgSCL (45 mg, 0.3739 mmol). The mixture was stirred at room température ovemight. The mixture was filtered, concentrated, and quenched with saturated sodium bicarbonate (50 mL) and extracted with DCM (75 mL). Additional DCM (4 x 50 mL) was used to wash the aqueous layer, and then the combined organic layer was dried with MgSO4, filtered, and concentrated to yield crude intermediate ((6-methyl-4-oxopiperidin-2yl)methyl benzoate) C104, which was used in the next step without further purification.

To the C104 mixture from the first step was added 2-(5-chloro-3-thienyl)ethanol S2 (50 pL, 0.4043 mmol), MsOH (100 pL, 1.541 mmol) and DCM (1 mL). The mixture was stirred at 40 °C ovemight. The mixture was pH adjusted to around pH 10, and then the mixture was phase separated. Purification by silica gel chromatography (Gradient: 0-20% MeOH in DCM) yielded [(2'S,6^,S,7S)-2-chloro-6^,-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4^,-piperidine]-2^,yl]methyl benzoate C105 (12 mg, 7%). *H NMR (300 MHz, Chloroformé δ 8.15 - 7.94 (m, 2H), 7.60 (t, J= 7.4 Hz, 1H), 7.48 (t, J= 7.6 Hz, 2H), 6.60 (s, 1H), 4.37 (dd, J= 10.9, 4.2 Hz, 1H), 4.18 (dd, J= 10.9, 8.0 Hz, 1H), 3.93 (t, J= 5.5 Hz, 2H), 3.61 - 3.44 (m, 1H), 3.21 (ddd, J= 11.4, 6.3, 2.4 Hz, 1H), 2.62 (t, J= 5.5 Hz, 2H), 2.09 (td, J= 13.3, 2.5 Hz, 2H), 1.57 - 1.34 (m, 2H), 1.14 (d, J = 6.3 Hz, 3H). LCMS m/z 392.02 [M+H]⁺.

277

Step 2. Synthesis of (2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-2 '-yl)methanol (210)

To a mixture of C105 (10 mg, 0.02552 mmol) in methanol (1 mL) was added NaOH (100 pL of 2 M, 0.2000 mmol) and the mixture was stirred at room température for five minutes. Purification by reversed-phase HP LC (Method: Cl 8 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H₂O with 5 mM HCl) yielded (2-chloro-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-2'-yl)methanol (Hydrochloride sait) 210 (5.5 mg, 66%). ‘H NMR (300 MHz, DMSOé₆) δ 8.83 (s, 1H), 8.33 (s, 1H), 6.93 (s, 1H), 3.90 (d, J= 5.5 Hz, 2H), 3.66 (dd, J= 11.8, 3.9 Hz, 1H), 3.53 (dd, J= 11.6, 5.3 Hz, 1H), 3.34 (s, 2H), 2.58 (t, J = 5.4 Hz, 2H), 2.15 (t, J= 14.4 Hz, 3H), 1.78 (q, J= 12.8 Hz, 2H), 1.27 (d, J= 6.5 Hz, 3H). LCMS m/z 288.14 [M+H]⁺.

Préparation ofS47 (2S, 6S)-2-cyclopropyl-6-methyl-piperidin-4-one (S47)

S25

L-Proline TEA

MgSO₄

O

S47

To a mixture of (4S)-4-aminopentan-2-one (Hydrochloride sait) S25 (150 mg, 1.090 mmol) in Et₃N (153 pL, 1.098 mmol) and EtOH (9 mL) was added cyclopropanecarbaldehyde (90 pL, 1.204 mmol), L-proline (25 mg, 0.2171 mmol), and MgSO₄ (130 mg, 1.080 mmol). The mixture was stirred at room température for 2 hours. The mixture was filtered, concentrated, and diluted in saturated aqueous bicarbonate (3 mL), diluted with water (7 mL) and extracted with DCM (2x10 mL). The combined organic layer was washed with brine (5 mL), dried with MgSO₄, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-20% MeOH in DCM) afforded (2S,6S)-2-cyclopropyl-6-methyl-piperidin-4-one S47 (65 mg, 39%). ‘H NMR (300 MHz, Chloroformé) δ 2.79 (dqd, J= 12.2, 6.1, 3.0 Hz, 1H), 2.35 (ddd, J= 14.0, 3.1, 2.1 Hz, 1H), 2.27 - 2.13 (m, 2H), 2.02 (ddd, J= 14.0, 11.6, 1.1 Hz, 1H), 1.96 - 1.85 (m, 1H), 1.80 (s, 1H), 1.12 (d, J= 6.2 Hz, 3H), 0.78 (qt, J= 8.2, 4.9 Hz, 1H), 0.59 - 0.27 (m, 2H), 0.23 - 0.03 (m, 2H). LCMS m/z 154.05 [M+l]⁺.

278

Préparation of Intermediates S48-S53

Intermediates S48-S53 were prepared in a single step from intermediate S25 or S24 using the appropriate aldéhyde and the method described for intermediate S47. Aldéhydes were prepared by methods described above or obtained from commercial sources. In this method, partial stereochemical érosion of the enantiomerically pure starting material (4S)-4-aminopentan2-one (Hydrochloride sait) S25 was observed, leading to unseparated mixtures of stereoisomers where the cis-product was the major isomer. Any modifications to methods are noted in Table 9 and accompanying footnotes.

Table 9. Method of préparation, structure and physicochemical data for intermediates S48S53

Product	Aldéhyde Reagent And Amine	Method	²hnmr
S48 0	Si c F	15; \\ A	Préparation ofS47^1,2	Ή NMR (300 MHz, Chloroform-7) δ 3.03 - 2.85 (m, 1H), 2.73 - 2.51 (m, 1H), 2.53 - 2.37 (m, 1H), 2.37 - 2.19 (m, 2H), 2.19-1.98 (m, 1H), 1.74 (s, 1H), 1.73 - 1.58 (m, 1H), 1.49 (ttd, J = 12.1, 8.0, 4.2 Hz, 1H), 1.22 (d, J = 6.2 Hz, 3H), 1.18-1.03 (m, 1H). LCMS m/z 190.09 [M+H]⁺.
S49 0 A	S24; J	Préparation ofS47’·²·³	'H NMR (300 MHz, Chloroformé) δ 2.92 (dqd, J= 12.3, 6.2, 2.9 Hz, 1H), 2.61 (ddd, 7=11.8, 6.0,2.8 Hz, 1H), 2.41 - 2.25 (m, 2H), 2.12 -1.98 (m, 2H), 1.69 (dq, J= 13.3, 6.7 Hz, 1H), 1.20 (d, J= 6.2 Hz, 3H), 0.94 (dd, J= 9.2, 6.8 Hz, 6H).
S50 A H	. S24; Q \\	Préparation ofSd/¹·²·³	Ή NMR (300 MHz, Chloroform-7) δ 2.98 (dqd, J= 12.2, 6.2, 2.9 Hz, 1H), 2.77 (ddt, J= 12.5, 6.4, 3.3 Hz, 1H), 2.36 (ddt, J= 14.4, 9.9, 2.4 Hz, 2H), 2.15-1.98 (m, 2H), 1.69 -1.41 (m, 2H), 1.22 (d, J= 6.2 Hz, 3H), 0.95 (t, J= 7.5 Hz, 3H).
S51 0 AL^L^^ <U ^H	S24; (D'A	Préparation ofS47^1,2,3	Ή NMR (300 MHz, Chloroform-7) δ 4.08 - 3.85 (m, 2H), 3.35 (td, J= 11.8, 2.2 Hz, 2H), 2.92 (dqd, J= 12.2, 6.2, 2.8 Hz, 1H), 2.65 (ddd, J= 11.9, 6.3, 2.8 Hz, 1H), 2.35 (ddt, 7= 14.4, 11.6, 2.4 Hz, 2H), 2.12 -1.97 (m, 2H), 1.73 - 1.63 (m, 1H), 1.63 1.52 (m, 2H), 1.46 - 1.30 (m, 2H), 1.19 (d, 7= 6.2 Hz, 3H).

279

Product	Aldéhyde Reagent And Amine	Method	Ή NMR
S52 V H	S24; J	Préparation ofS47’’²·³	Ή NMR (300 MHz, Chlorofonn-7) δ 2.89 (dqd, J= 12.2, 6.1, 3.0 Hz, 1H), 2.45 (ddd, 7= 14.0, 3.1, 2.2 Hz, 1H), 2.36 - 2.23 (m, 2H), 2.13 (ddd, 7= 13.9, 11.6, 1.0 Hz, 1H), 2.00 1.93 (m, 1H), 1.22 (d, 7= 6.2 Hz, 3H), 0.89 (qt, 7= 8.3, 5.0 Hz, 1H), 0.61 - 0.46 (m, 2H), 0.20 (qq, 7 = 8.0,3.9,3.4 Hz, 2H).
O S53^ ^H	S24; (D \\	Préparation ofS47^4,5	Ή NMR (300 MHz, Chloroform-7) δ 2.94 (dqd, 7= 12.3, 6.1, 2.7 Hz, 1H), 2.75 (ddd, 7= 11.7, 8.8, 2.7 Hz, 1H), 2.69 (s, 1H), 2.37 - 2.23 (m, 3H), 2.07 (ddd, 7= 13.8, 11.3,7.9 Hz, 2H), 2.01 -1.83 (m, 3H), 1.83 1.61 (m, 3H), 1.19 (d, 7= 6.2 Hz, 3H). LCMS m/z 167.95 [M+H]⁺.

1. The reaction was stirred ovemight.

2. Purification by silica gel chromatography (Gradient: 0-10% MeOH in DCM) afforded the product.

3. After completion the mixtures were concentrated, re-diluted in DCM and filtered.

4. After completion the mixture was poured into saturated aqueous sodium bicarbonate (10 mL) and diluted with water (3 mL) and ethyl acetate (30 mL).

5. The reaction was not purified, telescoped to the next step.

Compound 211 (2S, 6S)-2-chloro-2 '-cyclopropyl-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4'-piperidine]

S47 211

To a mixture of (2S,6S)-2-cyclopropyl-6-methyl-piperidin-4-one S47 (65 mg, 0.4242 mmol) in DCM (2 mL) was added 2-(5-chloro-3-thienyl)ethanol (75 pL) followed by MsOH (130 pL, 2.003 mmol) and the mixture was refluxed for 40 minutes. The mixture was cooled, pH adjusted with NaOH (500 pL of 6 M, 3.000 mmol) pH 14 and the organic layer was separated and concentrated. Purification by silica gel chromatography (Gradient: 0-20% MeOH in DCM) 280 afforded the freebase. The freebase was re-diluted in diethyl ether and HCl (100 pL of 4 M in dioxane, 0.4000 mmol), at which point a white solid precipitated. The mixture was concentrated to yield (2S,6S)-2-chloro-2'-cyclopropyl-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine] 211 (hydrochloride sait) (15.2 mg, 11%). *H NMR (300 MHz, MethanolA) δ 6.74 (s, 1H), 3.92 (t, J= 5.5 Hz, 2H), 3.65 - 3.51 (m, 1H), 2.75 (ddd, J= 12.6, 9.8, 3.0 Hz, 1H), 2.63 (dd, 7= 6.1, 4.9 Hz, 2H), 2.35 (ddt,7=23.2, 14.7, 2.8 Hz, 2H), 1.85 (dd,7= 14.6, 12.3 Hz, 1H), 1.70 (dd, 7= 14.6, 12.2 Hz, 1H), 1.36 (d, 7= 6.6 Hz, 3H), 0.93 (ddt, 7= 13.1, 9.5, 4.2 Hz, 1H), 0.81 - 0.64 (m, 2H), 0.64 - 0.53 (m, 1H), 0.44 - 0.26 (m, 1H). LCMS m/z 298.05 [M+H]⁺.

Compounds 212-218

Compounds 212-218 (see Table 10) were prepared from a single Oxa-Pictet Spengler step with isolated piperidone (see Table 9) and S2 as described for compound 211. Thiophene éthanol and piperidones were prepared by methods described above or obtained from commercial sources. As previously described in the préparation of the piperidinone intermediates, partial stéréo Chemical érosion of the enantiomerically pure starting material (4S)4-aminopentan-2-one (Hydrochloride sait) S25 was observed, leading to unseparated mixtures of stereoisomers where the cis-product was the major isomer. Any modifications to methods are noted in Table 10 and accompanying footnotes.

Table 10. Method of préparation, structure and physicochemical data for compounds 212-218.

Product	Piperidone	Method	'H NMR; LCMS m/z [M+H]⁺
Compound 212 oA£V^{c\| s} AA N V H	O ^pvA S48	Compound 211¹·^2,3	Ή NMR (300 MHz, DMSOA) δ 9.00 (d, 7= 23.3 Hz, 2H), 3.90 (d, 7= 4.6 Hz, 2H), 3.51 (s, 1H), 3.09 (s, 1H), 2.57 (d, 7= 5.4 Hz, 2H), 2.24 (d, 7= 15.6 Hz, 2H), 1.94 (d, 7= 13.2 Hz, 2H), 1.75 (d, 7= 13.3 Hz, 2H), 1.57 (s, 1H), 1.27 (d, 7= 6.6 Hz, 3H). LCMS m/z 334.15 [M+H]⁺.
Compound 213 oOCAci AA n V H	O ^fvX S48	Compound 211¹·²·³	‘H NMR (300 MHz, DMSOA) δ 8.92 (s, 1H), 8.70 (s, 1H), 3.96 3.82 (m, 2H), 3.45 (s, 1H), 3.21 (s, 1H), 2.59 (s, 2H), 2.31 -2.24 (m, 1H), 2.21 (s, 1H), 2.08 (d, J= 13.1 Hz, 2H), 1.76 (dd, 7= 28.8, 15.3 Hz, 4H), 1.27 (d, 7= 6.5 Hz, 3H). LCMS m/z 333.96 [M+H]⁺.

281

Product	Piperidone	Method	Ή NMR; LCMS m/z [M+H]⁺
Compound 214 ₀O3^^ci H 1	Â S49	Compound 211⁴	Ή NMR (300 MHz, Chloroform7)5 6.58 (s, 1H), 3.91 (t,7=5.5 Hz, 2H), 3.10 (dddd, J= 12.6, 8.9, 6.4, 2.5 Hz, 1H), 2.71 (ddd, J = 11.5, 6.8, 2.4 Hz, 1H), 2.60 (t, J = 5.5 Hz, 2H), 2.10 (dt, J= 13.5, 2.5 Hz, 1H), 2.02 (dt, 7= 13.6, 2.6 Hz, 1H), 1.58 (dt,7= 13.4, 6.7 Hz, 2H), 1.31 (ddd, 7= 13.5, 11.4, 4.3 Hz, 2H), 1.10 (d, 7 = 6.4 Hz, 3H), 0.94 (dd, 7= 12.7, 6.7 Hz, 6H). LCMS m/z 300.08 [M+H]⁺.
Compound 215 θόΑ X	0 A H S50	Compound 211⁴	^!H NMR (300 MHz, Chloroform-7) 5 6.58 (s, 1H), 3.92 (t, 7= 5.5 Hz, 2H), 3.16 (dt, 7=9.4,3.6 Hz, 1H), 3.00-2.85 (m, 1H), 2.61 (t, 7= 5.5 Hz, 2H), 2.14- 1.96 (m, 2H), 1.49-1.42 (m, 2H), 1.41 - 1.26 (m, 3H), 1.13 (d, 7= 6.4 Hz, 3H), 0.95 (t, J= 7.5 Hz, 3H). LCMS m/z 286.04 [M+H]⁺.
Compound 216 oOA Aç s ^H Y·⁰	0 <U ^H S51	Compound 211⁴	‘H NMR (300 MHz, Chloroformd) 5 6.56 (s, 1H), 4.07 - 3.92 (m, 2H), 3.89 (t, 7= 5.5 Hz, 2H), 3.35 (tt, 7= 11.7, 2.6 Hz, 2H), 3.07 (dtd, 7= 12.6, 6.3, 2.4 Hz, 1H), 2.76 (ddd, 7= 11.5, 6.9, 2.4 Hz, 1H), 2.58 (td, 7= 5.4, 1.3 Hz, 2H), 2.06 (ddt, 7= 20.3, 13.5, 2.5 Hz, 2H), 1.69 (dt, 7= 12.4, 2.6 Hz, 1H), 1.64- 1.59 (m, 1H), 1.52 - 1.29 (m, 4H), 1.26 (dd,7= 10.2,3.1 Hz, 1H), 1.08 (d,7=6.3 Hz, 3H). LCMS m/z 342.06 [M+H]⁺.
Compound 217 oOA^CIΛ A	Â S52	Compound 211⁴	Ή NMR (300 MHz, Chloroformd) δ 6.56 (s, 1H), 3.87 (t, 7= 5.5 Hz, 2H), 3.03 (dtd, 7= 12.5, 6.3, 2.5 Hz, 1H), 2.58 (t, 7= 5.5 Hz, 2H), 2.22-2.02 (m, 2H), 1.98 (dt, 7= 13.7, 2.5 Hz, 1H), 1.54 (dd, 7= 13.3, 11.1 Hz, 1H), 1.37 (dd, 7=13.6,11.3 Hz, 1H), 1.10 (d, 7= 6.3 Hz, 3H), 0.77 (ddq, 7 = 13.2, 8.5, 4.8 Hz, 1H), 0.55 0.36 (m, 2H), 0.26 - 0.04 (m, 2H). LCMS m/z 298.1 [M+H]⁺.

282

Product	Piperidone	Method	'II NMR; LCMS m/z [M+H]⁺
Compound 218 <OlV^cix<A^s	O		Ή NMR (300 MHz, Chloroformez) δ 6.56 (s, 1H), 3.91 (t, 7=5.5 Hz, 2H), 3.08 (q, 7= 4.9, 4.2 Hz, 1H), 2.96 - 2.75 (m, 1H), 2.59 (t,
	Compound	J= 5.5 Hz, 2H), 2.28 - 2.04 (m,
	/7 VJ H	2 11⁴·⁵	2H), 2.08-1.81 (m, 4H), 1.74 (dq, 7=16.8, 8.3, 7.8 Hz, 3H),
H VJ	S53		1.42- 1.23 (m, 2H), 1.16 (d, 7 = 13.4 Hz, 1H), 1.08 (d, 7= 6.4 Hz, 3H). LCMS m/z 312.05 [M+H]⁺.

1. The reaction was pH adjusted with saturated aqueous sodium bicarbonate, the organic layer was separated, and the solvent was removed in vacuo.

2. Purification by reversed-phase HPLC (Method: C18 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H₂O with 5 mM HCl) afforded the product.

3. Compounds 212 and 213 were separated during purification.

4. The compound was not salted with HCl after column chromatography

5. Reaction was run for 5 minutes

Compound 219 (2S, 6R)-2-chloro-2 '-methyl-6'-(tetrahydropyran-4-ylmethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] (219)

S25 MgSO₄

To a vial of 2-tetrahydropyran-4-ylacetaldehyde (23.99 mg, 0.1872 mmol), MgSO4 (35 mg, 0.2908 mmol) and L-proline (5 mg, 0.04343 mmol) was added a solution of (4S)-4aminopentan-2-one (Hydrochloride sait) S28 (28 mg, 0.1872 mmol)) in EtOH (1 mL). Triethylamine (30 pL, 0.2152 mmol) was added to each vial. The reaction was stirred at room température over the weekend. The reaction mixture was evaporated under a stream of nitrogen. A solution of 2-(5-chloro-3-thienyl)ethanol (25 pL, 0.2075 mmol) in dioxane (750 pL) was added. A solution of triflic acid (100 pL, 1.130 mmol) in dioxane (750 pL) was added. The resulting mixture was stirred at room température for 30 minutes. The reaction mixture was quenched with NaOH (2.0 mL of 2 M, 4.000 mmol) and DCM (1.5 mL), and the resulting biphasic mixtures were stirred for several minutes. The mixture was passed through a 25 pM

283 polypropylene filter plate, and the resulting DCM layers were isolated and evaporated. Purification by reversed-phase HP LC (Method: C18 Waters Sunfire column (30 x 150 mm, 5 micron). Gradient: MeCN in H2O with 0.1% trifluoroacetic acid) yielded (2S,6R)-2-chloro-2'methyl-6^,-(tetrahydropyran-4-ylmethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] (Trifluoroacetate sait) 219 (22.8 mg, 32%). *H NMR (400 MHz, DMSO-cfe) δ 8.56 (s, 1H), 8.23 (s, 1H), 6.93 (s, 1H), 3.90 (t, J= 5.5 Hz, 2H), 3.81 (d, J= 11.4 Hz, 2H), 3.55 - 3.20 (m, 4H), 2.62 - 2.55 (m, 2H), 2.24 (t, J= 14.0 Hz, 2H), 1.76 - 1.38 (m, 7H), 1.23 (d, J= 6.5 Hz, 3H), 1.28 1.03 (m, 2H). LCMS m/z 356.23 [M+H]⁺.

Préparation of S54 (2 S, 6S)-2 '-chloro-2-cyclopropyl-6-methyl-l-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'thieno[2,3-c]pyran]-4'-one (S54)

O

Step 1. Synthesis ofl-[(2S,6S)-2-chloro-2'-cyclopropyl-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-1 ’-yl]-2,2,2-trifluoro-ethanone (Cl 06)

To a mixture of (2'S,6'S,7S)-2-chloro-2'-cyclopropyl-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] 211 (Hydrochloride sait) (30 mg, 0.08974 mmol) in DCM (1 mL) was added DIPEA (50 pL, 0.2871 mmol) followed by trifluoroacetic anhydride (15 pL, 0.1079 mmol). The reaction was stirred for 2 hours. The mixture was quenched with 1 N HCl, separated, and concentrated. Purification by silica gel chromatography (Gradient: 0-30% EtOAc in Heptane) afforded l-[(2S,6S)-2-chloro-2'-cyclopropyl-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C106 (31 mg, 87%). Ή NMR (300 MHz, Chloroform-J) δ 6.59 (s, 1H), 4.60 - 3.95 (m, 1H), 3.82 (t, J= 5.5 Hz, 2H), 3.25 (s, 1H), 2.57 (t, J= 5.5 Hz, 2H), 2.50 (dd, J= 16.1, 8.1 Hz, 2H), 2.07 (d, J= 15.1 Hz, 2H), 1.62 -1.19 (m, 3H), 1.18 - 0.85 (m, 1H), 0.86 - 0.38 (m, 3H), 0.32 (dq, J= 9.2, 4.7 Hz, 1H). LCMS m/z 394.04 [M+H]⁺. The product is a mixture of two rotamers based on NMR.

284

Step 2. Synthesis of (2S,6S)-2'-chloro-2-cyclopropyl-6-methyl-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one (S54)

To a mixture of l-[(2S,6S)-2-chloro-2'-cyclopropyl-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C106 (30 mg, 0.07617 mmol) in acetonitrile (500 pL) was added N-hydroxyphthalimide (8 mg, 0.04904 mmol) and cobaltous diacetate tetrahydrate (2 mg, 0.008029 mmol). The mixture was vacuum purged with an oxygen balloon three times. The mixture was heated to 45 °C and stirred. After 6 hours, the reaction was cooled to room température. The mixture was vacuum purged with nitrogen three times and then diluted with MTBE (3 mL) and saturated sodium bicarbonate (3 mL). The layers were separated, and the organic layer was washed with water (2x2 mL) and brine (20 mL). The organic layer was dried with NazSCU, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-30% EtOAc in heptane) afforded (2S,6S)-2'-chloro-2-cyclopropyl6-methyl-l-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one (13 mg, 41%) S54. LCMS m/z 408.0 [M+H]⁺.

Compound 220 (2'S,6'S, 7S)-2-chloro-2'-cyclopropyl-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-4-ol) (226)

Step 1. Synthesis of l-[(2S,6S)-2-chloro-2'-cyclopropyl-4-hydroxy-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4’-piperidine]-1 '-yl]-2,2,2-trifluoro-ethanone (C107)

To a solution of (2S,6S)-2'-chloro-2-cyclopropyl-6-methyl-1-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S54 in MeOH (0.5 mL) was added NaBH₄ (1 mg, 0.02643 mmol). After 1 hour, the mixture was diluted with MTBE (5 mL), washed with saturated brine, and concentrated to give crude l-[(2'S,6'S,7S)-2-chloro-2'cyclopropyl-4-hydroxy-6'-methyl-spiro [4,5 -dihydrothieno [2,3 -c]pyran-7,4'-piperidine] -1 -yl] 2,2,2-trifluoro-ethanone C107.

285

Step 2. Synthesis of (2S,6S)-2-chloro-2'-cyclopropyl-6'-methyl-spiro[4,5-dihydrothieno[2,3c/pyran- 7,4 ’-piperidine]-4-ol (220)

Crude l-[(2'S,6'S,7S)-2-chloro-2'-cyclopropyl-4-hydroxy-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C107 was diluted with NaOH (500 pL of 6 M, 3.000 mmol) followed by MeOH (0.5 mL) and the mixture was stirred at 50 °C. After 40 minutes, the mixture was cooled to room température, diluted with DCM (5 mL) and pH adjusted to pH 10 with saturated aqueous ammonium chloride. The organic phase was separated, passed through a phase separator, and concentrated. Purification by silica gel chromatography (Gradient: 0-20% MeOH in DCM) afforded (2S,6S)-2-chloro-2'-cyclopropyl-6'methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol 220 (6 mg, 24%). *H NMR (300 MHz, Chloroformé δ 6.60 (s, 1H), 4.20 (q, J= 3.1 Hz, 1H), 3.74 (dd, J= 12.3, 2.9 Hz, 1H), 3.63 (dd, 7= 12.3, 2.8 Hz, 1H), 3.01 - 2.71 (m, 1H), 2.12 -1.70 (m, 4H), 1.46 (t, J= 12.2 Hz, 1H), 1.37-1.19 (m, 1H), 1.19- 1.02 (m, 1H), 0.92 (dd, 7= 6.3, 3.4 Hz, 3H), 0.61 (t,7= 10.9 Hz, 1H), 0.28 (dt, J= 9.9, 6.2 Hz, 2H), -0.08 (dd, 7= 9.0, 4.4 Hz, 2H). LCMS m/z 314.07 [M+H]⁺.

Compound 221 (2S, 4S, 6S)-2 '-chloro-2-ethynyl-6-methyl-45'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]

C113

· TMEDA, sec-BuLi, DN1F

^C114 C11S

Step 1. Synthesis of tert-butyl (S)-7-methyl-l,4-dioxa-8-azaspiro[4.5]decane-8-carboxylate To a solution of tert-butyl (2S)-2-methyl-4-oxo-piperidine-l-carboxylate C113 (40 g, 183.80 mmol) in toluene (1000 mL) were added ethylene glycol (22.916 g, 21 mL, 361.82 mmol) and PPTS (7.5 g, 29.248 mmol). The mixture was refluxed for 72 hours under Dean-Stark conditions. The mixture was then cooled to room température, quenched with saturated sodium bicarbonate (300 mL), and extracted with EtOAc (2 x 500 mL). The combined organic layer was dried over Na2SÛ4 and concentrated. Purification by silica gel column chromatography (Gradient: 0-5% EtOAc:Heptane) afforded tert-butyl (7S)-7-methyl-l,4-dioxa-8azaspiro[4.5]decane-8-carboxylate C114 (35 g, 67%) as a yellow liquid. *H NMR (300 MHz, 286

CDC1₃) δ 4.49-4.44 (m, 1H), 4.03-3.88 (m, 5H), 3.12-3.02 (m, 1H), 1.90-1.83 (m, 1H), 1.68-1.56 (m, 3H), 1.46 (s, 9H), 1.23 (d, J = 6.9 Hz, 3H), LCMS m/z 258.31 [M+l]⁺.

Step 2. Synthesis of tert-butyl (7S,9S)-7-ethynyl-9-methyl-l ,4-dioxa-8-azaspiro[4.5] decane8-carboxylate (Cl 15)

i. To a solution of tert-butyl (S)-7-methyl-l,4-dioxa-8-azaspiro[4.5]decane-8-carboxylate C114 (14 g, 53.862 mmol) in diethyl ether (210 mL) was added TMEDA (7.5460 g, 10 mL, 63.638 mmol) at room température. The reaction was cooled to -78°C, and then sec-Butyllithium (60 mL of 1.4 M, 84.000 mmol) was added slowly. The reaction was stirred at -78°C for 2 hours. DMF (7.8635 g, 8.5 mL, 105.43 mmol) was added slowly while maintaining température at -78°C and stirred for 1 hour. The reaction was then warmed to room température and stirred for 1 hour. The mixture was quenched with saturated ammonium chloride solution (100 mL) and extracted with ethyl acetate (2 x 200 mL). The combined organic layer was concentrated. Purification by silica gel chromatography (Gradient: 0-50% EtOAc:Heptane) to yield crude tertbutyl (9S)-7-formyl-9-methyl-l,4-dioxa-8-azaspiro[4.5]decane-8-carboxylate (20 g, 65%) as a pale yellow liquid. The intermediate did not ionize by LCMS, and material was a mixture of diastereomers as observed by *H NMR, so the crude product was carried forward to the next step.

ii. To the crude product from the previous step dissolved in MeOH (300 mL) was added l-diazo-l-dimethoxyphosphoryl-propan-2-one (17 g, 75.217 mmol) and K?CO₃ (15 g, 106.36 mmol) at room température. The reaction mixture was stirred for 16 hours at room température. The reaction mixture was filtered and washed with EtOAc (50 mL). The filtrâtes were concentrated. Purification by silica gel column chromatography (0-5% EtOAc:Heptane) provided the isolated single stereoisomer tert-butyl (7S,9S)-7-ethynyl-9-methyl-l,4-dioxa-8azaspiro[4.5]decane-8-carboxylate C115 (9.4 g, 59%) as a colorless liquid. *H NMR (400 MHz, DMSO-d6) δ : 5.032-5.014 (m, 1H), 4.23-4.19 (m, 1H), 3.97-3.93 (m, 2H), 3.84-3.80 (m, 2H), 3.11-3.10 (m, 1H), 1.95-1.78 (m, 4H), 1.39 (s, 9H), 1.35 (d, J = 6.8 Hz, 3H), LCMS m/z 282.35 [M+l]⁺

Step 3. Synthesis of (2'S,6'S,7S)-2-chloro-2'-ethynyl-6'-methyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] (221)

To a mixture of tert-butyl (7S,9S)-7-ethynyl-9-methyl-l,4-dioxa-8-azaspiro[4.5]decane8-carboxylate C115 (25 mg, 0.08886 mmol) and 2-(5-chloro-3-thienyl)ethanol (20 pL, 0.1986 mmol) in DCM (0.5 mL) was added MsOH (40 pL, 0.6164 mmol). The mixture was heated to 40 °C for 2 hours. The mixture was allowed to cool to room température and then pH adjusted with saturated aqueous bicarbonate and aqueous 1 N NaOH to pH 11. Additional DCM (2 mL) 287 was added, and the organic layer was separated and concentrated. Purification by silica gel chromatography (Gradient: 0-10% MeOH:DCM) yielded (2'S,6'S,7S)-2-chloro-2'-ethynyl-6'methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] 221 (14 mg, 50%) as a yellow oil. ’H NMR (300 MHz, Methanol-d4) δ 6.67 (s, 1H), 3.90 (t, J = 5.5 Hz, 2H), 3.84 (dt, J = 11.8, 2.6 Hz, 1H), 3.05 (dqd, J = 13.0, 6.4, 2.6 Hz, 1H), 2.68 (d, J = 2.3 Hz, 1H), 2.59 (t, J = 5.5 Hz, 2H), 2.20 (dt, J = 13.8, 2.7 Hz, 1H), 1.97 (dt, J =13.8, 2.6 Hz, 1H), 1.71 (dd, J =13.8, 11.8 Hz, 1H), 1.32 (dd, J = 13.8, 11.5 Hz, 1H), 1.07 (d, J = 6.4 Hz, 3H). LCMS m/z 282.09 [M+l]⁺ NMR analysis by NOE confirmed the 2,4-trans 2,6-cis stereochemistry.

Préparation of S55 (2S,4S, 6S)-2'-chloro-2-ethynyl-6-methyl-l-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'thieno[2,3-c]pyran] -4'(5'H)-one

221

TFAA, DIPEA

Co(ll) acetate tetrahydrate N-hydroxypthalimide, O₂ balloon

C116

S55

Step 1. Synthesis of l-[(2S,6'S,7S)-2-chloro-2'-ethynyl-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-1 ’-yl]-2,2,2-trifluoro-ethanone (Cl 16)

A solution of (2'S,6'S,7S)-2-chloro-2'-ethynyl-6'-methyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] 221 (6.91 g, 24.5 mmol) in DCM (100 mL) was cooled to 0 °C. DIPEA (8.5 mL, 48.80 mmol) was added, followed by TFAA (4.3 mL, 30.93 mmol), and the mixture was stirred for 1 minute. The mixture was quenched with 1 N HCl (50 mL), and the organic layer separated. The organic layer was washed with aqueous 1 N HCl (50 mL), dried with MgSCU, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-5% EtOAc:heptane) provided the crude product as a crystalline white-pink solid. This material was suspended in TBME (28 mL) and stirred at reflux, which fully dissolved the solid. The solution was cooled to 0 °C and stirred for 15 minutes. The resulting solid was filtered and rinsed with cold TBME. The resulting mother liquor was concentrated, and the recrystallization process was repeated twice to provide three crops of white solid. The combined crops were concentrated to yield l-[(2'S,6'S,7S)-2-chloro-2'-ethynyl-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-l'-yl]-2,2,2-trifluoro-ethanone C116 (7.42 g, 80%). *H NMR (300 MHz, Chloroform-d) δ 6.60 (s, 1H), 5.19 (s, 1H), 4.40 (p, J = 7.4, 6.9 Hz, 1H), 3.83 (td, J = 5.6, 2.2 Hz,

288

2H), 2.68 - 2.52 (m, 4H), 2.37 (dt, J = 23.7, 14.2 Hz, 3H), 1.51 (d, J = 6.6 Hz, 3H). LCMS m/z 378.07 [M+l]⁺

Step 2. Synthesis of 1-[(2'S,6'S, 7S)-2-chloro-2'-ethynyl-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone (S55)

To a solution of l-[(2'S,6'S,7S)-2-chloro-2'-ethynyl-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-T-yl]-2,2,2-trifluoro-ethanone C116 (5.0 g, 13.18 mmol) in MeCN (50 mL) was added N-hydroxyphthalimide (3.0 g, 18.39 mmol) and cobalt(II) acetate tetrahydrate (330 mg, 1.325 mmol). The mixture was vacuum purged with an oxygen balloon six times and oxygen balloon placed into septum to maintain oxygen atmosphère. The mixture was heated to 55 °C and stirred ovemight. The mixture was cooled to room température, then vacuum purged with nitrogen three times and diluted with MTBE (200 mL) and saturated bicarbonate (100 mL). The layers were separated, and the organic layer was washed with saturated bicarbonate (4 x 100 mL). Brine (100 mL) was added and the layers were separated. The organic layer was dried with MgSCU, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-10% EtOAc:heptane) yielded (2S,4S,6S)-2'-chloro-2-ethynyl-6methyl-l-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S55 (2.67 g, 52%) as a white crystalline solid. Ή NMR (300 MHz, Chloroform-d) δ 7.19 (s, 1H), 5.26 (d, J = 21.5 Hz, 1H), 4.38 (tt, J = 11.1, 5.8 Hz, 1H), 4.26 (t, J = 1.2 Hz, 2H), 2.77 (dd, J = 15.2, 8.3 Hz, 1H), 2.65 - 2.45 (m, 3H), 2.43 - 2.27 (m, 1H), 1.53 (d, J = 6.5 Hz, 3H).

Compound 222 (2 'S, 4S, 6'S, 7S)-2-chloro-2 '-ethynyl-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 piperidine]-4-ol (222)

S55 C117 222

Step 1. Synthesis of l-[(2'S,4S,6'S,7S)-2-chloro-2'-ethynyl-4-hydroxy-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-l’-yl]-2,2,2-trifluoro-ethanone (Cil7)

To a solution of N-[(lR,2R)-2-amino-l,2-diphenyl-ethyl]-4-methyl-benzenesulfonamide (80 mg, 0.2183 mmol) in MeCN (15 mL) was added dichloro(pentamethylcyclopentadienyl)

289 rhodium(III) dimer (65 mg, 0.1035 mmol). The mixture was stirred at room température for 5 minutes, then 5:2 formic acid-triethylamine complex (3 mL, 7.144 mmol) was added. After 5 minutes, the mixture was cooled to 0°C, and (2S,4S,6S)-2'-chloro-2-ethynyl-6-methyl-1-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S55 (2670 mg, 6.809 mmol) in MeCN (70 mL) was added. The mixture was stirred at 0°C for 2 hours and then allowed to warm to room température and stirred 72 hours. The mixture was diluted with ethyl acetate (20 mL) and then washed with 1 N HCl (20 mL), saturated sodium bicarbonate (20 mL), and brine (20 mL). The organic layer was dried with MgSCU, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-40% EtOAc:heptane) yielded l-[(2'S,4S,6'S,7S)-2-chloro-2'ethynyl-4-hydroxy-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2trifluoro-ethanone C117 (2.27 g, 85%) as a white solid. 'H NMR (300 MHz, Chloroform-d) δ 6.85 (s, 1H), 5.19 (s, 1H), 4.48 (t, J = 9.2 Hz, 2H), 3.86 (qd, J = 12.5, 3.1 Hz, 2H), 2.75 - 2.46 (m, 3H), 2.37 (dd, J = 14.6, 6.8 Hz, 1H), 2.20 (d, J = 14.7 Hz, 1H), 2.01 (d, J = 9.1 Hz, 1H), 1.53 (d, J = 6.6 Hz, 3H). LCMS m/z 394.13 [M+l]⁺

Step 2. Synthesis of (2 'S, 4S, 6'S, 7S)-2-chloro-2'-ethynyl-6'-methyl-spiro[4,5dihydrothieno]2,3-c]pyran-7,4 ’-piperidine]-4-ol (222)

To a solution of l-[(2'S,4S,6'S,7S)-2-chloro-2^,-ethynyl-4-hydroxy-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2- trifluoro-ethanone C117 (1.5 g, 3.809 mmol) in MeOH (7.5 mL) was added NaOH (4 mL of 6 M, 24.00 mmol). The mixture was stirred for 15 minutes at room température, then filtered and rinsed with additional water and concentrated to yield (2'S,4S,6'S,7S)-2-chloro-2'-ethynyl-6'-methyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-4-ol 222 (1.07 g, 88%) as a white solid. ^!H NMR (300 MHz, Chloroform-d) δ 6.84 (s, 1H), 4.44 (d, J = 7.8 Hz, 1H), 4.04 - 3.80 (m, 3H), 3.51 (s, 2H), 3.21 (dd, J = 11.4, 5.7 Hz, 1H), 2.35 (d, J = 14.1 Hz, 1H), 2.05 (d, J = 9.3 Hz, 1H), 2.00 - 1.85 (m, 1H), 1.74 - 1.63 (m, 1H), 1.45 (dd, J = 13.4, 11.3 Hz, 1H), 1.13 (d, J = 6.3 Hz, 3H). LCMS m/z 298.07 [M+l]⁺.

290

Compound 223 (2 'S, 6'S, 7S)-2-chloro-2'-ethynyl-4,4-difluoro-6'-methyl-spiro[5H-thieno[2,3-c]pyran-7,4'piperidine] (223)

S55 C118 223

Step 1. Synthesis of l-[(2'S,6'S,7S)-2-chloro-2'-ethynyl-4,4-difluoro-6'-methyl-spiro[5Hthieno[2,3-c]pyran- 7,4 ’-piperidine]-1 ’-yl]-2,2,2-trifluoro-ethanone (Cl 18)

A mixture of (2S,4S,6S)-2'-chloro-2-ethynyl-6-methyl-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S55 (1690 mg, 4.314 mmol) in DAST (5 mL, 37.84 mmol) was heated to 40 °C and stirred ovemight. Another portion of DAST (5 mL, 37.84 mmol) was added, and the reaction was heated to 40 °C for 36 hours. The mixture was cooled to room température and added dropwise into a stirring solution of saturated sodium bicarbonate (25 mL) maintained at 0 °C. The mixture was diluted with DCM (25 mL) and layers separated. The aqueous layer was extracted with additional DCM (25 mL), and the combined organic layer was dried with MgSCU, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-40% ETOAc:heptane) yielded l-[(2'S,6'S,7S)-2-chloro-2'-ethynyl4,4-difluoro-6'-methyl-spiro[5H-thieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoroethanone C118 (1.34 g, 73%) as an orange oil. *H NMR (300 MHz, Chloroform-d) δ 6.99 (s, 1H), 5.29 (d, J = 44.5 Hz, 1H), 4.41 (q, J = 7.7 Hz, 1H), 3.99 (t, J = 10.0 Hz, 2H), 2.70 (dd, J = 15.2, 8.3 Hz, 1H), 2.60 (s, 1H), 2.50 (t, J = 8.4 Hz, 2H), 2.32 (d, J = 15.3 Hz, 1H), 1.53 (d, J = 6.5 Hz, 3H). LCMS m/z 414.07 [M+l]⁺

Step 2. Synthesis of (2'S,6'S, 7S)-2-chloro-2'-ethynyl-4,4-difluoro-6'-methyl- spiro[5Hthieno]2,3-c]pyran- 7,4 ’-piperidine] (223)

To a mixture of l-[(2'S,6^,S,7S)-2-chloro-2'-ethynyl-4,4-difluoro-6'-methyl-spiro[5Hthieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C118 (1.36 g, 3.287 mmol) in MeOH (25 mL) was added NaOH (2.5 mL of 6 M, 15.00 mmol). The reaction was heated to 50 °C for 1 hour. The mixture was cooled to room température and diluted with TB ME (30 mL) and water (30 mL). The organic layer was washed with brine (50 mL) and dried with MgSÜ4, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-10%

291

MeOH:DCM) yielded (2'S,6'S,7S)-2-chloro-2'-ethynyl-4,4-difluoro-6'-methyl- spiro[5Hthieno[2,3-c]pyran-7,4'-piperidine] 223 (970 mg, 84%) as a brown oil. *H NMR (300 MHz, Methanol-d4) δ 7.07 (s, 1H), 4.10 (t, J = 10.4 Hz, 2H), 3.85 (dd, J = 11.7, 3.0 Hz, 1H), 3.12 2.97 (m, 1H), 2.77 - 2.68 (m, 1H), 2.30 (dt, J = 13.8, 2.9 Hz, 1H), 2.16 - 2.02 (m, 1H), 1.76 (dd, J = 13.7, 11.8 Hz, 1H), 1.44- 1.30 (m, 1H), 1.10 (d, J = 6.4 Hz, 3H). LCMS m/z 317.99 [M+l]⁺

Préparation of C 120

N-diazosulfamoyl fluoride

sodium azide water/MeCN/MTBE 0 °C oj?

F'%, 'Nt 'N'

C119 C120

Synthesis of N-diazosulfamoyl fluoride (C120)

To a mixture of NaN? (1.95 g, 30.00 mmol) in water (60 mL) cooled to 0 °C was added MTBE (60 mL) followed by an MeCN (3 mL) solution of 2,3-dimethylimidazol-3-ium-lsulfonyl fluoride (Trifluoromethanesulfonate) (11.8 g, 35.95 mmol) C119. After stirring vigorously for 10 minutes, the layers were allowed to separate for 30 minutes and the organic layer was removed, passed over a phase separator and placed in a conical flask, and aged ovemight. At this time, the formed red solution at the bottom of the flask was removed with a pipet and the organic layer was diluted with DMSO (60 mL). A biphasic mixture was observed. Acetonitrile (about 25 mL) was added until a homogenous mixture was observed. The final total volume was about 150 mL. This reaction mixture was used in subséquent azide formations without purification or characterization. The yield was estimated to be about 90% of active reagent based on literature precedent*. Based on this, the concentration was estimated to be about 0.18 M.

292

Compound 224 (2 'S, 4S, 6'S, 7S)-2-chloro-2 '-[l-[[4-(hydroxymethyl)phenyl] methyl] triazol-4-yl]-6'-methylspiro[4,5-dihydrothieno[2,3-c]pyra.n- 7,4 '-piperidine]-4-ol (224)

Examples:

C120

C121

KHCO₃water/DMSO/MeCN/MTBE

C123

OH

citric acid DMSO, water 50 °C

Step 1. Synthesis of [4-(azidomethyl)phenyl]methanol (C123)

To a mixture of [4-(aminomethyl)phenyl]methanol C121 and aqueous KHCO3 (400 pL of 3 M) was added an MTBE/DMSO/MeCN solution of N-diazosulfamoyl fluoride C120 (1.7 mL of 0.18 M) and the reaction was stirred at room température for 2 hours. At this time, reactions were assumed to be complété based on literature precedent^x and were not further characterized and [4-(azidomethyl)phenyl]methanol C123 was used directly in the next step.

Step 2. Synthesis of (2'S,4S,6'S,7S)-2-chloro-2'-[l-[[4-(hydroxymethyl)phenyl]methyl]triazol4-yl]-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 '-piperidine]-4-ol (224)

With continuai rapid stirring, approximately a third of the formed biphasic suspension (700 pL) from Step 1 was added to a solution of (2'S,4S,6'S,7S)-2-chloro-2'-ethynyl-6'-methylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol 222 in DMSO (0.1 mL), 350 pL of aqueous pH 5 buffer (sodium ascorbate (350 pL of 0.125 M):disodium phosphate:citric acid 1:4:2, 0.125 M based on sodium ascorbate) and 100 pL of an aqueous solution of CuSO₄ (100 pL of 0.035 M):3-[4-[[bis[[l-(3-hydroxypropyl)triazol-4-yl]methyl]amino]methyl]triazol-lyl]propan-l-ol 1:1 was added and the mixture was stirred at 50 °C ovemight. At this time, the reaction was cooled to room température and then dried in vacuo to remove highly volatile solvents. The resulting solution or suspension was then passed over a fîlter membrane for purification by reversed-phase HPLC. (Method: Waters XSelect CSH Cl8 OBD Prep Column; 19 x 100 mm, 5 micron. Gradient: Acetonitrile in Water with 10 mM Ammonium Hydroxide).) The product-containing fractions were pooled and concentrated to yield (2'S,4S,6'S,7S)-2-chloro2'-[l-[[4-(hydroxymethyl)phenyl]methyl]triazol-4-yl]-6'-methyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-4-ol 224 (3.5 mg, 21%) as a yellow solid. ’H NMR (400 MHz, DMSOd6) δ 7.97 (s, 1H), 7.30 (t, J = 5.7 Hz, 4H), 6.96 (s, 1H), 5.50 (s, 2H), 5.39 (s, 1H), 5.20 (s, 1H),

293

4.47 (s, 2H), 4.38 (t, J = 4.5 Hz, 1H), 4.10 (d, J = 11.5 Hz, 1H), 3.92 (dd, J = 11.9, 4.2 Hz, 1H), 3.63 (dd, J = 11.7, 5.1 Hz, 1H), 3.07 (s, 1H), 2.27 (s, 1H), 2.17 (d, J = 13.3 Hz, 1H), 2.03 (d, J = 13.3 Hz, 1H), 1.54 (t, J = 12.5 Hz, 1H), 1.23 (t, J = 12.3 Hz, 1H), 1.00 (d, J = 6.3 Hz, 3H). LCMS m/z 461.12 [M+l]⁺

Compound 225 (4-((4-((28,4S,68)-2'-chloro-4',4'-difluoro-2-methyl-4', 5'-dihydrospiro[piperidine-4,7'thieno[2,3-c]pyran]-6-yl)-lH-l,2,3-triazol-l-yl)methyl)phenyl)methanol (225)

C121

C120

KHCO3 --------------------->

water/D MS O/M eC N/MTB E

223

CuSO₄

THPTA sodium ascorbate disodium phosphate citric acid DMSO, water 50 °C

225

Step 1. Synthesis of [4-(azidomethyl)phenyl]methanol (C123)

To a mixture of [4-(aminomethyl)phenyl]methanol C121 and aqueous KHCO3 (400 pL of 3 M) was added an MTBE/DMSO/MeCN solution of N-diazosulfamoyl fluoride C120 (1.7 mL of 0.18 M), and the reaction was stirred at room température for 2 hours. At this time, reactions were assumed to be complété based on literature precedent^x and were not further characterized and [4-(azidomethyl)phenyl]methanol C123 was used directly in the next step.

Step 2. Synthesis of (4-((4-((28,4S,6S)-2'-chloro-4',4'-difluoro-2-methyl-4',5'dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-6-yl)-lH-l,2,3-triazol-lyl)methyl)phenyl)methanol (225)

With continuai rapid stirring, approximately a third of the formed biphasic suspension (700 pL) from Step 1 was added to a solution of (2'S,6'S,7S)-2-chloro-2'-ethynyl-4,4-difluoro-6'methyl- spiro[5H-thieno[2,3-c]pyran-7,4'-piperidine] 223 in DMSO (0.1 mL), 350 pL of aqueous pH 5 buffer (sodium ascorbate (350 pL of 0.125 M):disodium phosphate:citric acid 1:4:2, 0.125 M based on sodium ascorbate) and 100 pL of an aqueous solution of CuSO₄ (100 pL of 0.035 M) : 3 - [4-[ [bis [ [ 1 -(3 -hydroxypropyl)triazol-4-yl]methyl] amino]methyl]triazol-1 -yl]propan-1 -ol 1:1 was added and the mixture was stirred at 50 °C ovemight. At this time, the reaction was cooled to room température and then dried in vacuo to remove highly volatile solvents. The resulting solution or suspension was then passed over a filter membrane for purification by reversed-phase HP LC. (Method: Waters XSelect CSH Cl 8 OBD Prep Column; 19 x 100 mm, 5

294 micron. Gradient: Acetonitrile in Water with 10 mM Ammonium Hydroxide.) The productcontaining fractions were pooled and concentrated to yield (4-((4-((2S,4S,6S)-2'-chloro-4',4'difluoro-2-methyl-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-6-yl)-lH-l,2,3-triazoll-yl)methyl)phenyl)methanol 225 (3.5 mg, 21%) as a yellow solid. Ή NMR (400 MHz, DMSO5 d6) δ 8.00 (s, 1H), 7.34 (s, 1H), 7.29 (d, J = 2.3 Hz, 4H), 5.52 (s, 2H), 5.20 (s, 1H), 4.47 (d, J =

4.6 Hz, 2H), 4.21 (t, J = 10.6 Hz, 2H), 4.11 (d, J = 11.5 Hz, 1H), 3.02 (d, J = 18.8 Hz, 1H), 2.33 (d, J = 12.7 Hz, 2H), 2.10 (d, J = 13.4 Hz, 1H), 1.66 (dd, J = 13.5, 11.7 Hz, 1H), 1.30 (dd, J = 13.6, 11.3 Hz, 1H), 1.01 (d, J = 6.2 Hz, 3H). LCMS m/z 481.12 [M+l]⁺

Compounds 226-371

Compounds 226-371 (see Table 11) were prepared as either parent or trifluoroacetate salts following the methods described for compounds 224 and 225. 222 or 223 and appropriate amine were used, where the amine is converted in situ to an azide for cycloaddition to the desired triazole.

Table 11. Method of préparation, structure, and physicochemical data for compounds

226-371

Product	Starting Material and Amine	NMR, LCMS
Compound 226 OH °Ô>^CI N-J HO^Z O—	222; nh₂L 0 /	Ή NMR (400 MHz, DMSO-d6) δ 7.87 (s, 1H), 6.96 (s, 1H), 5.40 (d, J = 6.3 Hz, 1H), 5.28 (d, J = 5.6 Hz, 1H), 4.43 - 4.34 (m, 2H), 4.27 4.17 (m, 1H), 4.12 (dd, J = 11.3, 2.5 Hz, 1H), 3.93 (dd, J = 11.8,4.3 Hz, 2H), 3.64 (dd, J= 11.9, 5.2 Hz, 1H), 3.27 (s, 3H), 3.25 (d, J = 5.6 Hz, lH),3.10(d, J = 8.5 Hz, 1H), 2.18 (d, J= 13.5 Hz, 1H), 2.05 (d, J = 13.3 Hz, 1H), 1.58 (t, J = 12.5 Hz, 1H), 1.31-1.20 (m, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 429.15 (M+l)⁺
Compound 227	222;	Ή NMR (400 MHz, DMSO-d6) δ 7.86 (d, J = 1.6 Hz, 1H), 6.96 (s, 1H), 5.40 (s, 1H), 4.41 -

295

Product	Starting Material and Amine	NMR, LCMS
'^Z/--\ °~λ = X )-o \ Z\__/ t * ^ωχ·χ g	NH, S	4.23 (m, 3H), 4.11 (d, J = 11.2 Hz, 1H), 3.93 (dd, J = 11,8, 4.2 Hz, 1H), 3.84 (d, J = 11.4 Hz, 1H), 3.64 (dd, J = 11.9,5.2 Hz, 2H), 3.29 (s, 1H), 3.09 (s, 1H), 2.28 (s, 1H), 2.18 (d, J = 13.4 Hz, 1H), 2.05 (d, J = 13.3 Hz, 1H), 1.77 (s, 1H), 1.57 (t, J =11.8 Hz, 2H), 1.43 (d, J= 10.2 Hz, 3H), 1.31 - 1.12 (m, 2H), 1.02 (d, J = 6.3 Hz, 3H). LCMS m/z 439.17 (M+l)⁺
Compound 228 OH ÔA^CI N-J 0 \	222; nh₂ A°	Ή NMR (400 MHz, DMSO-d6) δ 8.01 (s, 1H), 6.97 (s, 1H), 5.41 (s, 1H), 4.44 (t, J = 7.0 Hz, 2H), 4.43 - 4.36 (m, 1H), 4.12 (d, J= 11.4 Hz, 1 H), 3.93 (dd, J = 11.8, 4.3 Hz, 1H), 3.64 (dd, J = 11.8, 5.2 Hz, 1H), 3.15-3.06 (m,3H), 2.99 (s, 3H), 2.28 (s, 1H), 2.22 (dt, J = 20.0, 9.8 Hz, 3H), 2.06 (d, J = 13.3 Hz, 1H), 1.58 (t, J = 12.5 Hz, 1H), 1.30-1.20 (m, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 461.12 (M+l)⁺
Compound 229 OH oOq*-⁰¹ -f-yx' N-J 0^__ o—	222; nh₂ O^__ o—	'H NMR (400 MHz, DMSO-d6) δ 7.94 (s, 1H), 6.96 (s, 1H), 5.42 (s, 1H), 4.46 (t, J = 5.3 Hz, 2H), 4.39 (t, J = 4.6 Hz, 1H), 4.12 (d, J = 11.4 Hz, 1 H), 3.93 (dd, J = 11.8, 4.3 Hz, 1H), 3.79 (t, J = 5.2 Hz, 2H), 3.64 (dd, J =11.8, 5.3 Hz, 1H), 3.51 (dd, J = 5.9, 3.5 Hz, 2H), 3.40 (dd, J = 5.7,3.6 Hz, 2H), 3.21 (s, 3H), 3.10 (s, 1H),

296

Product	Starting Material and Amine	NMR, LCMS
		2.24 (s, 1H), 2.17 (d, J = 13.4 Hz, 1H), 2.05 (d, J = 13.3 Hz, 1H), 1.56 (dd, J= 13.4, 11.6 Hz, 1H), 1.30- 1.20 (m, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 443.15 (M+l)⁺
Compound 230 OH oÔl>“ N-J X NH / θ' '	222; nh₂ X NH / .H	Ή NMR (400 MHz, DMSO-d6) δ 7.88 (d, J = 7.5 Hz, 2H), 6.97 (s, 1H), 5.42 (s, 1H), 4.40 (t, J = 4.8 Hz, 1H), 4.36 (t, J = 6.1 Hz, 2H), 4.11 (d, J = 11.4 Hz, 1H), 3.92 (dd, J = 11.8, 4.3 Hz, 1H), 3.64 (dd, J = 11.7, 5.4 Hz, 1H), 3.44 (d, J = 6.1 Hz, 1 H), 3.24 (t, J = 5.4 Hz, 1H), 3.21 (s, 0H), 3.10 (s, 1H), 2.28 (p, J = 7.0 Hz, 1H), 2.19 (s, lH),2.16(d, J = 13.5 Hz, 1H), 2.06 (d, J = 13.4 Hz, 1H), 1.56 (t, J = 12.4 Hz, 1H), 1.291.16 (m, 1H), 1.02 (d, J = 6.3 Hz, 3H), 0.94 (dd, J = 6.9, 1.5 Hz, 6H). LCMS m/z 454.17 (M+l)⁺
Compound 231 OH °ÔQ-^ci y	222; NH₂o \v —s—(^z v h \—/ 0 —'	Ή NMR (400 MHz, DMSO-d6) δ 8.06 (s, 1H), 8.01 (dd, J = 7.9, 1.4 Hz, 1H), 7.76 - 7.67 (m, 1H), 7.64 (t, J = 7.9 Hz, 1H), 7.10 (d, J = 7.6 Hz, 1H), 6.96 (s, 1H), 6.03 (s, 2H), 5.42 (s, 1H), 4.39 (t, J = 4.8 Hz, 1H), 4.14 (d, J= 11.5 Hz, 1H), 3.94 (dd, J = 11.9, 4.2 Hz, 1H), 3.64 (dd, J = 11.8, 5.2 Hz, lH),3.30(s, 3H), 3.09 (s, 1H), 2.33 (s, 1H), 2.21 (d, J = 13.4 Hz, 1H), 2.05 (d, J =13.3

297

Product	Starting Material and Amine	NMR, LCMS
		Hz, 1H), 1.63 - 1.52 (m, 1H), 1.31 - 1.16 (m, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 509.12 (M+l)⁺
Compound 232 OH o0q*-^c\| N-⁴ N H	222; nh₂ho N H	‘H NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.69 (d, J = 8.1 Hz, 1H), 7.52 (d, J = 8.4 Hz, 1H), 7.34 (dd, J = 8.3, 6.9 Hz, 1H), 7.10 (t, J = 7.5 Hz, 1H), 6.95 (s, 1H), 5.90 (s, 2H), 4.37 (t, J = 4.7 Hz, 1H), 4.08 (d, J =11.5 Hz, 1H), 3.91 (dd, J= 11.9, 4.2 Hz, 1H), 3.61 (dd, J = 11.9, 5.2 Hz, 1H), 3.05 (s, 1H), 2.24 (s, 1H), 2.15 (d, J= 13.3 Hz, 1H), 2.02 (d, J= 13.1 Hz, 1H), 1.58 - 1.47 (m, 1H), 1.22 (t, J= 12.2 Hz, 1H), 0.98 (d, J = 6.3 Hz, 3H). LCMS m/z 471.14 (M+l)⁺
Compound 233 OH oÔT>^cl	222; nh₂V	Ή NMR (400 MHz, DMSO-d6) δ 7.93 (s, 1H), 6.97 (s, 1H), 5.49 (s, 2H), 4.40 (t, J = 4.7 Hz, 1H), 4.35 (s, 2H), 4.13 (d, J = 11.7 Hz, 1H), 3.94 (dd, J= 11.8, 4.2 Hz, 1H), 3.69 - 3.62 (m, 1 H), 3.11 (s, 2H), 2.26 (s, 1H), 2.21 (d, J = 13.4 Hz, 1H), 2.05 (d, J = 13.3 Hz, 1H), 1.651.52 (m, 1H), 1.27 (dd, J = 13.5, 11.3 Hz, 1H), 1.03 (d, J = 6.3 Hz, 3H), 0.76 - 0.63 (m, 3H). LCMS m/z 411.14 (M+l)⁺
Compound 234	222;	’H NMR (400 MHz, DMSO-d6) δ 8.69 (s, 1H), 8.06 (s, 1H), 7.55 (d, J = 3.5 Hz, 1H), 6.96

298

Product	Starting Material and Amine	NMR, LCMS
ο °*\ X ²¹ 5 { (Γ\ 'ρ=\ γ^ζ ^Ζ'' '^Ζ^Μ< JJ ζ ζ	nh₂ nfXk-NH \=N	(s, 1 H), 6.25 (d, J = 3.5 Hz, 1H), 5.94 (s, 2H), 5.41 (s, 1H), 4.39 (t, J = 4.5 Hz, 1H), 4.14 (d, J = 11.5 Hz, 1H), 3.94 (dd, J = 11.8,4.3 Hz, 1H), 3.64 (dd, J =11.9, 5.2 Hz, 1H), 3.10 (s, 1H), 2.29 (s, 1H), 2.20 (d, J= 13.4 Hz, 1H), 2.05 (d, J = 13.3 Hz, 1H), 1.64-1.53 (m, 1H), 1.25 (t, J= 12.2 Hz, 1H), 1.02 (d, J = 6.4 Hz, 3H). LCMS m/z 472.14 (M+l)⁺
Compound 235 ΟΗ °ÔM^ci _ν?ν ... LÀ,. Ν-¹ > ^Ν=\	222; NH₂ > ^N=\	^!H NMR (400 MHz, DMSO-d6) δ 7.92 (s, 1H), 6.95 (d, J =14.4 Hz, 2H), 6.84 (s, 1H), 5.42 (s, 1H), 4.65 (t, J = 7.4 Hz, 2H), 4.43 - 4.36 (m, 1H), 4.09 (d, J = 11.4 Hz, 1 H), 3.92 (dd, J = 11.9, 4.2 Hz, 1H), 3.63 (dd, J =11.8, 5.3 Hz, 1H), 3.18 (t, J = 7.4 Hz, 2H), 3.08 (s, 1H), 2.39 (s, 3H), 2.20 (s, 3H), 2.14 (d, J= 13.4 Hz, 1H), 2.05 (d, J= 13.2 Hz, 1H), 1.54 (t, J= 12.5 Hz, 1H), 1.24 (t, J= 12.3 Hz, 1H), 1.02 (d, J = 6.3 Hz, 3H). LCMS m/z 474.21 (M+l)⁺
Compound 236	222; nh₂ O J	’H NMR (400 MHz, DMSO-d6) δ 8.74 (d, J = 2.1 Hz, 1H), 8.11 (s, 1H), 8.06 (d, J = 8.0 Hz, 1H), 7.91 (dd, J = 8.1, 2.2 Hz, 1H), 6.96 (s, 1H), 5.74 (s, 2H), 5.42 (s, 1H), 4.39 (t, J = 4.7 Hz, 1H), 4.12 (d, J = 11.5 Hz, 1H), 3.93 (dd, J = 11.8, 4.2 Hz, 1H), 3.63 (dd, J =11.8, 5.3 Hz, 1H), 3.09 (s, 1H), 2.31

299

Product	Starting Material and Amine	NMR, LCMS
ζ'ί''² /—\ O—\ 12 A >© \__/\__/ I * ^ωχΑ O ^—		(s, 1H), 2.19 (d, J = 13.4 Hz, 1H), 2.04 (d, J = 13.1 Hz, 1H), 1.61 - 1.50 (m, 1H), 1.24 (dd, J = 13.3, 11.4 Hz, 1H), 1.01 (d, J = 6.3 Hz, 3H). LCMS m/z 457.15 (M+l)⁺
Compound 237 OH °ÔA^C\| AA N-⁴ —,Q N	222; nh₂ —4 „o N	Ή NMR (400 MHz, DMSO-d6) δ 7.94 (s, 1H), 6.96 (s, 1H), 5.38 (s, 2H), 4.38 (t, J = 4.7 Hz, 1H), 4.10 (d, J = 11.5 Hz, 1 H), 3.92 (dd, J = 11.7, 4.2 Hz, 1H), 3.63 (dd, J =11.9, 5.2 Hz, 1H), 3.08 (s, 1H), 2.43 (s, 3H), 2.28 (s, 1H), 2.17 (d, J= 13.4 Hz, 1H), 2.14 (s, 3H), 2.04 (d, J= 13.3 Hz, 1H), 1.55 (t, J = 12.5 Hz, 1H), 1.22 (d, J =12.6 Hz, 1H), 1.01 (d, J = 6.3 Hz, 3H). LCMS m/z 450.19 (M+l)⁺
Compound 238 OH _οΟ> _NRA N-J	. 222; nh₂ ?s° 0' \	*H NMR (400 MHz, DMSO-d6) δ 8.08 (s, 1H), 7.99 - 7.89 (m, 2H), 7.54 (d, J = 8.2 Hz, 2H), 6.96 (s, 1H), 5.69 (s, 2H), 5.42 (s, 1H), 4.39 (t, J = 4.7 Hz, 1H), 4.12 (d, J= 11.4 Hz, 1H), 3.93 (dd, J= 11.9, 4.2 Hz, 1 H), 3.63 (dd, J = 11.8,5.1 Hz, 1H), 3.20 (s, 3H), 3.09 (s, 1H), 2.30 (s, 1H), 2.19 (d, J = 13.3 Hz, 1H), 2.04 (d, J = 13.3 Hz, 1H), 1.61 1.50 (m, 1H), 1.29-1.16 (m, 1H), 1.01 (d, J = 6.3

300

Product	Starting Material and Amine	NMR, LCMS
		Hz, 3H). LCMS m/z 509.12 (M+l)⁺
Compound 239 OH oÔQ-⁰¹ <rr N⁴0^0	222; nh₂	Ή NMR (400 MHz, DMSO-d6) δ 7.84 (s, 1H), 6.97 (s, 1H), 5.37 (s, 3H), 4.40 (t, J = 4.7 Hz, 1H), 4.13 (d, J = 11.7 Hz, 1 H), 3.94 (dd, J = 11.9, 4.2 Hz, 1H), 3.64 (dd, J = 11.9, 5.3 Hz, 1H), 3.48 - 3.40 (m, 4H), 3.10 (s, 1H), 2.24 (s, 1H), 2.19 (d, J= 13.5 Hz, 1H), 2.06 (d, J = 13.3 Hz, 1H), 1.64-1.53 (m, 1H), 1.58 (s, 4H), 1.45 (s, 2H), 1.31 - 1.16 (m, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 466.18 (M+l)⁺
Compound 240 OH °ÔQ~^ci -<yV^% N** V^OH \r	222; nh₂\z~oh LT	‘H NMR (400 MHz, DMSO-d6) δ 7.85 (s, 1H), 6.96 (s, 1H), 5.38 (s, 1H), 4.93 (s, 1H), 4.39 (t, J = 4.6 Hz, 1H), 4.33 (s, 2H), 4.12 (d, J = 11.4 Hz, 1 H), 3.93 (dd, J = 11.8, 4.2 Hz, 1H), 3.64 (dt, J = 12.2, 4.7 Hz, 3H), 3.54 (td, J = 7.9, 7.5, 3.6 Hz, 2H), 3.22 (s, 2H), 3.10 (s, 1H), 2.29 (s, lH),2.19(d, J= 13.4 Hz, 1 H), 2.05 (d, J = 13.2 Hz, 1H), 1.62-1.51 (m, 1H), 1.38-1.21 (m, 5H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 469.20 (M+l)⁺
Compound 241	222; nh₂ <r ^-S-NH > \ ⁰ //A	Ή NMR (400 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.26 (t, J = 7.7 Hz, 2H), 7.15 - 7.09 (m, 2H), 7.00 (t, J = 7.4 Hz, 1H), 6.96 (s, 1H), 5.40 (s, 1H), 4.67 (t, J = 7.1 Hz, 2H), 4.39 (s, 1H), 4.124.04 (m, 1H), 3.92 (dd, J

301

Product	Starting Material and Amine	NMR, LCMS
i T ''^z O=tozO ^==/ 1 \ \ O—< il Ί ^{1 Iz}v A \ / ^{T ω}χΆ O		= 11.9, 4.2 Hz, 1H), 3.68 - 3.61 (m, 1Ή), 3.61 (d, J = 7.2 Hz, 2H), 3.09 (d, J = 9.4 Hz, 1H), 2.13 (d, J = 13.4 Hz, 1H), 2.04 (d, J = 13.3 Hz, 1H), 1.52 (t, J = 12.5 Hz, 1H), 1.23 (t, J = 12.3 Hz, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 424.20 (M+l)⁺
Compound 242 OH oÔl>^c' <rr é ^j<JoH	222; nh₂ ^OH	‘H NMR (400 MHz, DMSO-d6) δ 7.93 (s, 1H), 6.97 (s, 1H), 5.41 (s, 1H), 4.74 (s, 1H), 4.39 (s, 1H), 4.28 (d, J = 1.4 Hz, 2H), 4.12 (d, J = 11.4 Hz, 1H), 3.94 (dd, J = 11.8, 4.2 Hz, 1H), 3.64 (dd, J = 11.8, 5.2 Hz, 1H), 3.11 (d, J = 4.6 Hz, 3H), 2.29 (s, 1H), 2.19 (d, J = 13.6 Hz, 1H), 2.05 (d, J= 13.3 Hz, 1H), 1.57 (dd, J= 13.3, 11.7 Hz, 1H), 1.31 - 1.16 (m, 1H), 1.03 (d, J = 6.3 Hz, 3H), 0.61 (q, J = 3.8 Hz, 2H), 0.48 (t, J = 2.9 Hz, 2H). LCMS m/z 425.18 (M+l)⁺
Compound 243 OH <0Q~^cl té N	222; NH, Vô	Ή NMR (400 MHz, DMSO-d6) δ 9.11 (d, J = 1.5 Hz, 1H), 8.61 (dd, J = 4.6, 1.6 Hz, 1H), 8.07 - 7.95 (m, 3H), 6.95 (s, 1H), 6.07 (s, 2H), 5.40 (s, 1H), 4.37 (t, J = 4.7 Hz, 1H), 4.08 (d, J = 11.3 Hz, 1H), 3.90 (dd, J = 11.9, 4.2 Hz, 1H), 3.61 (dd, J =11.9, 5.3 Hz, 1H), 3.06 (s, 1H), 2.27 (s, 1H), 2.15 (d, J = 13.5 Hz, 1H), 2.02 (d, J = 13.2 Hz, 1H), 1.57-1.46 (m, 1H), 1.27- 1.16 (m, 1H), 0.99 (d, J = 6.2 Hz,

302

Product	Starting Material and Amine	NMR, LCMS
		3H). LCMS m/z 472.18 (M+l)⁺
Compound 244 OH Ô>^CI N-⁴ HO'	222; nh₂ HO^Z	’H NMR (400 MHz, DMSO-d6) δ 7.92 (s, 1H), 6.96 (s, 1H), 5.40 (s, 1H), 5.05 (s, 1H), 4.39 (t, J = 4.8 Hz, 1H), 4.34 (t, J = 5.4 Hz, 2H), 4.12 (d, J = 11.4 Hz, 1 H), 3.93 (dd, J=11.9, 4.2 Hz, 1H), 3.75 (t, J = 5.5 Hz, 2H), 3.64 (dd, J = 11.9,5.3 Hz, 1H), 3.10 (s, 1H), 2.23 (s, 1H), 2.18 (d, J = 13.5 Hz, 1H), 2.05 (d, J = 13.3 Hz, 1H), 1.63 - 1.50 (m, 1H), 1.30- 1.20 (m, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 385.17 (M+l)⁺
Compound 245 OH <Ο> Λ'τ^% N-' F /°	222; nh₂ F 0 /	Ή NMR (400 MHz, DMSO-d6) δ 7.99 (s, 1H), 7.28 - 7.20 (m, 1H), 7.20-7.10 (m, 2H), 6.96 (s, 1H), 5.46 (s, 2H), 5.41 (s, 1H), 4.38 (t, J = 4.7 Hz, 1H), 4.10 (d, J = 11.5 Hz, 1H), 3.92 (dd, J = 11.8, 4.2 Hz, 1H), 3.82 (s, 3H), 3.63 (dd, J = 11.8,5.2 Hz, 1 H), 3.08 (s, 1H), 2.28 (s, 1H), 2.17 (d, J = 13.5 Hz, 1H), 2.03 (d, J= 13.3 Hz, 1H), 1.54 (t, J = 12.5 Hz, 1H), 1.28- 1.16 (m, 1H), 1.01 (d, J = 6.3 Hz, 3H). LCMS m/z 479.18 (M+l)⁺
Compound 246	222; HO—^NH₂HO	‘H NMR (400 MHz, DMSO-d6) δ 7.87 (s, 1H), 6.96 (s, 1H), 5.21 (s, 2H), 4.90 (s, 1H), 4.48 - 4.36 (m, 1H), 4.39 (s, 1H), 4.23 -4.08 (m, 2H), 3.93 (dd, J=11.9, 4.3 Hz, 1H), 3.79 (s,

303

Product	Starting Material and Amine	NMR, LCMS
τ O I I oj /--\ °~λ ιζ X >-o \__/\__/ I * o		1H), 3.64 (dd, J =11.8, 5.3 Hz, 1H), 3.29 (d, J = 6.4 Hz, 1H), 3.10 (s, 1H), 2.24 (s, 1H), 2.19 (d, J= 13.4 Hz, 1H), 2.05 (d, J= 13.2 Hz, 1H), 1.63 - 1.52 (m, 1H), 1.31 - 1.16 (m, 1H), 1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 415.15 (M+l)⁺
Compound 247 OH oôî>^cl N^'îî H	222; HO^ _zNH₂	'H NMR (400 MHz, DMSO-d6) δ 7.82 (s, 1H), 6.96 (s, 1H), 5.41 (s, 1H), 4.84 (s, 1H), 4.39 (t, J = 4.7 Hz, 1H), 4.21 (s, 2H), 4.12 (d, J = 11.7 Hz, 1H), 3.94 (dd, J = 11.9, 4.2 Hz, 1H), 3.64 (dd, J = 11.8, 5.2 Hz, 1H), 3.10 (s, 1H), 2.26 (s, 1H), 2.20 (d, J= 13.4 Hz, 1H), 2.05 (d, J = 13.3 Hz, 1H), 1.62-1.51 (m, 1H), 1.26 (dd, J = 13.3, 11.1 Hz, 1H), 1.08 - 1.00 (m, 8H). LCMS m/z 413.17 (M+l)⁺
Compound 248 OH oÔ>^cl N-'	222; nh₂ $\^.N	Ή NMR (400 MHz, DMSO-d6) δ 9.08 (s, 1H), 8.03 (s, 1H), 7.98 (s, 1H), 6.96 (s, 1H), 5.87 (s, 2H), 5.41 (s, 1H), 4.38 (t, J = 4.6 Hz, 1H), 4.10 (d, J= 11.6 Hz, 1 H), 3.92 (dd, J = 11.8, 4.3 Hz, 1H), 3.63 (dd, J = 11.9, 5.2 Hz, 1H), 3.08 (s, 1H), 2.29 (s, lH),2.17(d, J= 13.4 Hz, 1H), 2.04 (d, J = 13.3 Hz, 1H), 1.54 (dd, J = 13.3, 11.6 Hz, 1H), 1.29-1.16 (m, 1H), 1.01 (d, J = 6.2 Hz, 3H). LCMS m/z 438.14 (M+l)⁺

304

Product	Starting Material and Amine	NMR, LCMS
Compound 249 OH oôi>^cl/AA, N-^J N^OH	222; nh₂^^*nh N^OH	*H NMR (400 MHz, DMSO-d6) δ 7.94 (s, 1H), 6.98 - 6.85 (m, 4H), 5.47 (s, 2H), 4.38 (t, J = 4.7 Hz, 1H), 4.09 (d, J = 11.5 Hz, 1 H), 3.92 (dd, J = 11.7, 4.2 Hz, 1H), 3.62 (dd, J =11.9, 5.2 Hz, 1H), 3.07 (s, 1H), 2.25 (s, lH),2.17(d, J= 13.4 Hz, 1H), 2.03 (d, J = 13.3 Hz, 1H), 1.58-1.48 (m, 1H), 1.22 (t, J =12.4 Hz, 1H), 1.00 (d, J = 6.2 Hz, 3H). LCMS m/z 487.21 (M+l)⁺
Compound 250 OH O N-⁴	222; nh₂	'H NMR (400 MHz, DMSO-d6) δ 7.92 (s, 1H), 6.97 (s, 1H), 5.45 (d, J = 20.5 Hz, 1H), 4.39 (t, J = 4.7 Hz, 1H), 4.33 (d, J = 7.3 Hz, 2H), 4.11 (d, J= 11.3 Hz, 1H), 3.93 (dd, J = 11.9, 4.2 Hz, 1 H), 3.64 (dd, J = 11.9,5.2 Hz, 1H), 3.10 (s, 1H), 2.73 (h, J = 7.7 Hz, 1H), 2.27 (s, 1H), 2.22-2.14 (m, 1H), 2.05 (d, J= 13.3 Hz, 1H), 1.97 (qd, J = 7.3, 6.7, 4.4 Hz, 2H), 1.90- 1.70 (m, 4H), 1.57 (dd, J= 14.3, 10.7 Hz, 1H), 1.26 (dd, J = 13.3, 11.1 Hz, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 409.19 (M+l)⁺
Compound 251	222; nh₂ A	Ή NMR (400 MHz, DMSO-d6) δ 8.00 (t, J = 9.2 Hz, 1H), 7.06 (s, 1H), 6.98 (d, J = 13.7 Hz, 1H), 6.73 (s, 1H), 5.59 - 5.45 (m, 1H), 4.35 (d, J = 34.4 Hz, 4H), 4.13 (d, J= 12.1 Hz, 1H), 2.51 (s, 7H), 2.23 (d, J =12.7 Hz, 4H),

305

Product	Starting Material and Amine	NMR, LCMS
ο ¢- x /\/ \ ° Λ A ²¹ ²'' '^Z^ ^^Z<^ Z '•'Z		2.05 (d, J= 13.9 Hz, 1H), 1.58 (t, J =12.8 Hz, 1H), 1.27 (t, J = 12.9 Hz, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 463.19 (M+l)⁺
Compound 252 OH o0o~^ci <rr N-J “^OH	222; N H, >L	’H NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 6.96 (s, 1H), 5.49 (s, 1H), 4.62 (s, 1H), 4.41 -4.32 (m, 4H), 4.11 (d, J= 11.4 Hz, 1H), 3.92 (d, J = 4.0 Hz, 0H), 3.13 -3.07 (m, 1H), 2.17 (d, J = 13.5 Hz, 1H), 2.04 (d, J= 13.3 Hz, 1H), 1.94- 1.86 (m, 2H), 1.65 - 1.53 (m, 1H), 1.27 (t, J= 12.4 Hz, 1H), 1.12 (s, 6H), 1.02 (d, J = 6.3 Hz, 3H). LCMS m/z 427.21 (M+l)⁺
Compound 253 OH o0q~^ci<yÇ· N-J	222; nh₂	'H NMR (400 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.84 (d, J = 8.0 Hz, 2H), 7.44 (d, J = 8.0 Hz, 2H), 6.95 (s, 1H), 5.67 (s, 2H), 5.50 (s, 1H), 4.39 (s, 1H), 4.12 (dd, J = 11.7, 2.5 Hz, 1H), 3.93 (dd, J= 11.9, 4.1 Hz, 1H), 3.09 (dt, J = 9.0, 4.0 Hz, lH),2.19(dd, J = 13.2, 2.8 Hz, 1H), 2.03 (d, J = 13.6 Hz, 1H), 1.64-1.51 (m, 1H), 1.35 -1.21 (m, 1H), 1.01 (d, J = 6.3 Hz, 3H). LCMS m/z 456.15 (M+l)⁺
Compound 254	222;	‘H NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.47 (d, J = 3.3 Hz,

306

Product

Starting Material and Amine

NMR, LCMS

O

Compound 255

222;

OH

1H), 6.95 (s, 1H), 5.52 (d, J = 6.8 Hz, 1H), 4.39 (d, J = 4.9 Hz, 1H), 4.32 (d, J = 7.1 Hz, 2H), 4.12 (dd, J = 11.7, 2.6 Hz, 1H), 3.93 (dd, J= 12.0, 4.1 Hz, 1H), 3.10 (q, J = 5.7, 4.2 Hz, 1H), 3.00 (d, J = 10.2 Hz, 1H), 2.89 (t, J = 10.8 Hz, 1H), 2.32 2.10 (m,4H), 2.03 (d, J = 13.4 Hz, 1H), 1.68 (s, 1H), 1.58 (t, J = 12.6 Hz, 1H), 1.42 (ddd, J =10.5, 8.5,4.1 Hz, 1H), 1.341.23 (m, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 452.18 (M+l)⁺‘H NMR (400 MHz, DMSO-d6) δ 8.09 (s, 1H), 6.95 (s, 1H), 5.74 (s, 2H), 5.53 (s, 1H), 4.39 (d, J = 4.8 Hz, 1H), 4.14 (dd, J= 11.7, 2.5 Hz, 1 H), 3.94 (dd, J = 11.9, 4.1 Hz, 1H), 3.09 (d, J= 10.6 Hz, 1H), 2.20 (d, J = 13.5 Hz, 1H), 2.04 (d, J= 13.3 Hz, 1H), 1.59 (t, J = 12.6 Hz, 1H), 1.29 (t, J = 12.4 Hz, 1H), 1.03 (d, J = 6.3 Hz, 4H). LCMS m/z 380.12 (M+l)⁺_________

Ή NMR (400 MHz, DMSO-d6) δ 7.90 (s, 1H), 6.95 (s, 1H), 5.50 (s, 1H), 4.83 (s, 1H), 4.39 (t, J = 4.6 Hz, 1H), 4.34 (dd, J = 13.6, 6.0 Hz, 1H), 4.12 (dd, J = 13.4, 7.8 Hz, 2H), 3.93 (dd, J= 11.9, 4.1 Hz, 1H), 3.26 (t, J = 6.3 Hz, 2H), 3.10 (s, 2H), 2.33 (s, 1H), 2.19 (dt, J = 13.6, 2.6 Hz, 1H), 2.101.99 (m, 2H), 1.58 (t, J =

307

Product	Starting Material and Amine	NMR, LCMS
		12.6 Hz, 1H), 1.28 (dd, J = 13.4, 11.2 Hz, 1H), 1.03 (d,J = 6.3 Hz, 3H), 0.76 (d, J = 6.8 Hz, 3H). LCMS m/z 413.17 (M+l)⁺
Compound 257 OH οόί>“ N-^J	222; nh₂	*H NMR (400 MHz, DMSO-d6) δ 7.92 (s, 1H), 6.96 (s, 1H), 5.42 (s, 1H), 4.46 (t, J = 5.3 Hz, 2H), 4.39 (t, J = 4.7 Hz, 1H), 4.12 (d, J = 11.4 Hz, 1 H), 3.93 (dd, J = 11.9, 4.2 Hz, 1H), 3.74 (t, J = 5.3 Hz, 2H), 3.64 (dd, J = 11.9, 5.2 Hz, 1H), 3.47-3.39 (m, 2H), 3.09 (d, J = 9.9 Hz, 1H), 2.26 (s, 1H), 2.18 (d, J = 13.5 Hz, 1H), 2.05 (d, J = 13.4 Hz, 1H), 1.57 (t, J = 12.5 Hz, 1H), 1.26 (dd, J =13.4, 11.3 Hz, 1H), 1.10- 1.00 (m, 6H). LCMS m/z 413.17 (M+l)⁺
Compound 258 OH oÔl>^ci N-J ( HN— n	222; nh₂ K	Ή NMR (400 MHz, DMSO-d6) δ 7.89 (d, J = 4.9 Hz, 1H), 7.82 (d, J = 2.3 Hz, 1H), 6.96 (s, 1H), 5.42 (s, 1H), 4.43 (dt, J = 12.7, 6.3 Hz, 1H), 4.39 (d, J = 4.9 Hz, 1H), 4.25 (ddd, J= 13.7, 6.8, 2.3 Hz, 1H), 4.10 (d, J = 11.4 Hz, 1H), 3.93 (dd, J = 11.9, 4.2 Hz, 1H), 3.63 (dd, J= 11.8, 5.3 Hz, 1H), 3.09 (s, 1H), 2.89-2.81 (m, 1H), 2.53 (d, J = 4.2 Hz, 3H), 2.24 (s, lH),2.16(d, J = 13.4 Hz, 1H), 2.05 (d, J = 13.4 Hz, 1H), 1.54 (td, J = 13.0, 5.2 Hz, 1H), 1.23 (q, J =12.3 Hz, 1H), 1.02 (d, J = 6.3 Hz, 3H), 0.98 (d, J = 6.9 Hz,

308

Product	Starting Material and Amine	NMR, LCMS
		3H). LCMS m/z 440.21 (M+l)⁺
Compound 259 OH OQ^-01 '.^sAmA ^N. J H N —' OH	222; nh₂ OH	Ή NMR (400 MHz, DMSO-d6) δ 7.91 (s, ΊΗ), 7.16 (d, J = 8.4 Hz, 2H), 6.95 (s, 1H), 6.76 6.70 (m, 2H), 5.38 (s, 3H), 4.38 (t, J = 4.7 Hz, 1H), 4.08 (d, J= 11.4 Hz, 1 H), 3.92 (dd, J = 11.8, 4.2 Hz, 1H), 3.62 (dd, J =11.9, 5.2 Hz, 1H), 3.07 (s, 1H), 2.26 (s, lH),2.17(d, J= 13.4 Hz, 1H), 2.03 (d, J = 13.3 Hz, 1H), 1.60-1.48 (m, 1H), 1.23 (dd, J = 13.4, 11.2 Hz, 1H), 1.00 (d, J = 6.3 Hz, 3H). LCMS m/z 447.17 (M+l)⁺
Compound 260 OH °ÔO“^ci _K,ysA Ν'' ^h° ³~Ç~^	222; nh₂ ^h° ³~C~	Ή NMR (400 MHz, DMSO-d6) δ 7.92 (s, 1H), 7.19-7.11 (m, 1H), 6.97 (s, 1H), 6.74 (s, 3H), 6.72 (d, J = 8.3 Hz, 1H), 5.54 (s, 1H), 5.45 (s, 1H), 4.52 (dd, J = 13.9, 4.1 Hz, 1H), 4.39 (s, 2H), 4.18 (s, 1H), 4.12 (d, J = 11.5 Hz, 1H), 3.97-3.84 (m, 2H), 3.64 (dd, J= 11.9, 5.2 Hz, 1H), 3,09 (s, 1H), 2.26 (s, 3H), 2.23 (s, 1H), 2.17 (d, J= 13.3 Hz, 1H), 2.05 (d, J = 13.2 Hz, 1H), 1.57 (s, 1H), 1.30-1.16 (m, 1H), 1.02 (d, J = 6.3 Hz, 3H). LCMS m/z 505.19 (M+l)⁺
Compound 261	222;	*H NMR (400 MHz, DMSO-d6) δ 7.92 (d, J = 10.0 Hz, 1H), 7.47 (d, J = 7.3 Hz, 1H), 7.19 (ddt, J= 17.6, 10.0, 4.5 Hz, 3H), 6.97 (s, 1H),

309

Product	Starting Material and Amine	NMR, LCMS
x CL /χ ,Z Y z 'z NZ IZ Y )-o \__/\__/ x O	nh₂ >p	5.65 (s, 1H), 5.44 (s, 1H), 5.11 (s, 1H), 4.474.37 (m, 2H), 4.37 - 4.27 (m, 1H), 4.13 (d, J = 11.7 Hz, 1 H), 3.94 (dd, J = 11.9, 4.2 Hz, 1H), 3.64 (dd, J = 11.8, 5.3 Hz, 1H), 3.10 (s, 1H), 2.31 (d, J = 6.3 Hz, 3H), 2.24 (s, 1H), 2.18 (d, J = 15.4 Hz, 1H), 2.06 (d, J = 13.4 Hz, 1H), 1.57 (td, J = 12.4,6.6 Hz, 1H), 1.31 -1.16 (m, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 475.20 (M+l)⁺
Compound 262 OH oôjc t' N'^JA N—⁷	222; nh₂ N—⁷	Ή NMR (400 MHz, DMSO-d6) δ 9.10 (d, J = 1.4 Hz, 1H), 8.68 (d, J = 5.1 Hz, 1H), 7.96 (s, 1H), 7.39 (dd, J = 5.2, 1.5 Hz, 1H), 6.96 (s, 1H), 5.41 (s, 1H), 4.75 (t, J = 7.1 Hz, 2H), 4.39 (s, 1H), 4.08 (d, J =10.8 Hz, 1 H), 3.92 (dd, J = 11.8, 4.3 Hz, 1H), 3.63 (dd, J =11.8, 5.2 Hz, 1H), 3.34 (s, 2H), 3.08 (s, 1H), 2.23 (s, 1H), 2.14 (d, J= 13.4 Hz, 1H), 2.04 (d, J = 13.4 Hz, 1H), 1.58 - 1.48 (m, 1H), 1.29-1.19 (m, 1H), 1.01 (d, J = 6.2 Hz, 3H). LCMS m/z 447.17 (M+l)⁺
Compound 263	222; nh₂l·'	Ή NMR (400 MHz, DMSO-d6) δ 9.34 (s, 5H), 8.27 (s, 1H), 7.04 (s, 1H), 5.52 (d, J = 6.1 Hz, 1H), 5.50 (s, 2H), 4.73 (d, J = 12.3 Hz, 1H), 4.42 (q, J = 4.9 Hz, 1H), 4.00 (dd, J= 12.1, 4.0 Hz, 1H), 3.72 (s, 3H), 2.32 (d, J = 14.0 Hz, 1H), 2.19 (t, J =13.8

310

Product	Starting Material and Amine	NMR, LCMS
OH oÔl>^ci /N ν'*AjyA'/z, ^N. J H o^°		Hz, 1H), 1.82 (t, J =13.4 Hz, 1H), 1.28 (d, J = 6.5 Hz, 3H). LCMS m/z 413.12 (M+l)⁺
Compound 264 OH o0Q-^ci N-^J OH	222; nh₂ OH	’H NMR (400 MHz, DMSO-d6) δ 9.36 (s, 1H), 8.91 (s, 1H), 8.28 (s, 1H), 7.04 (s, 1H), 5.53 (d, J = 6.1 Hz, 1H), 4.68 (s, 1H), 4.58 (s, 1H), 4.44 (td, J = 8.0, 7.3,3.1 Hz, 3H), 3.99 (dd, J = 12.2, 4.0 Hz, 1H), 3.71 (dd, J= 12.1, 4.9 Hz, 1H), 3.64 (s, 1H), 3.26 (s, 2H), 2.45 (s, 1H), 2.33 (d, J = 14.0 Hz, 1H), 2.21 (t, J= 13.6 Hz, 1H), 1.94 (dt, J = 13.5, 6.8 Hz, 1H), 1.83 (dd, J= 14.3, 12.3 Hz, 1H), 1.58 (dq, J =14.0, 7.4 Hz, 1H), 1.51-1.41 (m, 1H), 1.28 (d, J = 6.4 Hz, 3H), 0.87 (d, J = 6.6 Hz, 3H). LCMS m/z 427.16 (M+l)⁺
Compound 265 OH ô>^cl N-⁴ J°	222; nh₂	‘H NMR (400 MHz, DMSO-d6) δ 9.32 (s, 1H), 8.92 (d, J = 11.3 Hz, 1H), 8.29 (s, 1H), 7.02 (d, J = 16.7 Hz, 1H), 5.53 (s, 1H), 4.70 (t, J = 11.2 Hz, 1H), 4.59 (d, J = 6.3 Hz, 2H), 4.42 (s, 1H), 3.99 (dd, J = 12.0, 4.0 Hz, 1H), 3.71 (dd, J = 12.1, 4.9 Hz, 1 H), 3.64 (s, 1 H), 3.283.20 (m, 1H), 3.18-3.03 (m, 2H), 3.02 - 2.85 (m, 2H), 2.46 (s, 1H), 2.33

311

Product	Starting Material and Amine	NMR, LCMS
		(d, J = 14.2 Hz, 1H), 2.29-2.18 (m, 1H), 2.15 (s, 1H), 1.84 (t, J = 12.5 Hz, 2H), 1.28 (d, J = 6.4 Hz, 3H). LCMS m/z 473.13 (M+l)⁺
Compound 266 OH ÔQ^^cinAj''T^%N-J θ~/°	222; nh₂ Q°	‘H NMR (400 MHz, DMSO-d6) δ 9.30 (s, 1H), 8.88 (s, 1H), 8.26 (s, 1H), 7.33 - 7.27 (m, 2H), 7.03 (s, 1H), 6.98 (d, J = 8.3 Hz, 2H), 5.55 (s, 2H), 5.50 (d, J = 6.2 Hz, 1H), 4.66 (d, J = 12.5 Hz, 1H), 4.56 (dt, J = 8.8, 4.5 Hz, 1H), 4.40 (d, J = 5.4 Hz, 1H), 3.98 (dd, J =12.0, 3.9 Hz, 1H), 3.88 - 3.78 (m, 1H), 3.83 (s, 1H), 3.69 (dd, J = 12.1, 4.8 Hz, 1H), 3.60 (s, 1H), 3.46 (td, J = 11.8, 10.7, 2.7 Hz, 2H), 2.30 (d, J = 14.4 Hz, 1H), 2.17 (t, J =13.5 Hz, 1H), 1.93 (d, J= 12.4 Hz, 2H), 1.81 (t, J= 13.3 Hz, 1H), 1.54 (dtd, J = 13.2,9.0, 4.0 Hz, 2H), 1.25 (d, J = 6.6 Hz, 3H). LCMS m/z 531.19 (M+l)⁺
Compound 267 OH oÔl>^ci N-J	222; nh₂	Ή NMR (400 MHz, DMSO-d6) δ 9.35 (d, J = 10.2 Hz, 1H), 8.92 (d, J =11.0 Hz, 1H), 8.24 (s, 1H), 7.03 (d, J = 14.3 Hz, 1H), 5.53 (s, 1H), 4.69 (t, J= 11.4 Hz, 1H), 4.42 (s, 1H), 4.33 (d, J = 7.0 Hz, 2H), 3.99 (dd, J = 12.0, 4.0 Hz, 1H), 3.83 (d, J = 11.6 Hz, 2H), 3.71 (dd, J= 12.0, 4.9 Hz, 1H), 3.64 (s, 1H), 3.24 (t, J =11.8 Hz, 2H), 2.45 (s, 1H), 2.33 (d, J = 14.0 Hz, 1H), 2.28-2.17

312

Product	Starting Material and Amine	NMR, LCMS
		(m, 1H), 1.84 (t, J = 13.3 Hz, 1H), 1.40 (d, J = 12.9 Hz, 2H), 1.28 (d, J = 6.6 Hz, 3H), 1.28 1.17 (m, 1H). LCMS m/z 439.22 (M+l)⁺
Compound 268 OH oÔl>^ci N-⁴ A	222; nh₂ < 0—V ⁰	*H NMR (400 MHz, DMSO-d6) δ 9.31 (s, 1H), 8.91 (d, J= 11.5 Hz, 1H), 8.19 (s, 1H), 7.04 (s, 1H), 6.88 (dd, J = 8.1, 1.7 Hz, 1H), 6.82 (t, J = 7.8 Hz, 1H), 6.73 (dd, J = 7.6, 1.7 Hz, 1H), 5.55 (s, 2H), 5.52 (d, J = 5.9 Hz, 1H), 4.70 (t, J = 11.3 Hz, 1H), 4.41 (d, J = 4.5 Hz, 1H), 4.34 4.22 (m, 4H), 3.98 (dd, J = 12.1, 4.0 Hz, 1H), 3.70 (dd, J = 12.0, 4.9 Hz, 1 H), 3.61 (s, 1H), 2.45 (s, 1H), 2.31 (d, J = 14.3 Hz, 1H), 2.25-2.14 (m, 1H), 1.83 (t, J = 13.3 Hz, 1H), 1.27 (d, J = 6.5 Hz, 3H). LCMS m/z 489.16 (M+l)⁺
Compound 269 OH A » N-J b.	222; nh₂b	‘H NMR (400 MHz, DMSO-d6) δ 9.36 (s, 1H), 8.93 (s, 1H), 8.30 (s, 1H), 7.04 (s, 1H), 5.53 (s, 1H), 4.69 (t, J = 10.9 Hz, 1H), 4.42 (d, J = 7.3 Hz, 3H), 3.99 (dd, J = 12.0, 4.0 Hz, 1H), 3.82 - 3.65 (m, 2H), 3.68 - 3.63 (m, 1H), 3.65 3.59 (m, 1H), 3.45 (dd, J = 8.8, 5.3 Hz, 1H), 2.72 (p, J = 6.4 Hz, 1H), 2.45 (s, 1H), 2.33 (d, J= 14.1 Hz, 1H), 2.28-2.16 (m, 1H), 1.99- 1.78 (m, 2H), 1.61 (dt, J =13.2, 6.6 Hz, 1H), 1.28 (d, J = 6.4 Hz, 3H). LCMS m/z 425.18 (M+l)⁺

313

Product	Starting Material and Amine	NMR, LCMS
Compound 270 OH Om^ci « N-⁷	222; nh₂	Ή NMR (400 MHz, DMSO-d6) δ 9.40 (s, 1H), 8.95 (s, 1H), 8.23 (s, 1H), 7.04 (s, 1H), 5.53 (d, J = 6.2 Hz, 1H), 4.71 (s, 1H), 4.42 (d, J = 5.4 Hz, 1H), 4.36 (s, 2H), 4.00 (dd, J= 12.1, 4.0 Hz, 1H), 3.71 (dd, J = 12.0, 4.9 Hz, 1H), 3.65 (s, 1H), 3.22 (s, 3H), 3.03 (s, 2H), 2.47 (s, 1H), 2.33 (d, J =14.1 Hz, 1H), 2.21 (t, J= 13.6 Hz, 1H), 1.84 (t, J= 13.3 Hz, 1H), 1.29 (d, J = 6.5 Hz, 3H), 0.73 (q, J = 4.3 Hz, 2H), 0.54 (t, J = 3.0 Hz, 2H). LCMS m/z 439.17 (M+l)⁺
Compound 271 OH ô>^ci DH	222; nh₂ DH	Ή NMR (400 MHz, DMSO-d6) δ 9.30 (d, J = 10.4 Hz, 1H), 8.88 (d, J = 11.2 Hz, 1H), 8.31 (s, 1H), 7.38 -7.24 (m, 3H), 7.21 (d, J = 7.5 Hz, 1H), 7.04 (s, 1H), 5.64 (s, 2H), 5.51 (s, 1H), 5.27 (s, 1H), 4.69 (t, J = 11.3 Hz, 1H), 4.48 (s, 2H), 4.41 (s, 1H), 3.98 (dd, J = 12.0,3.9 Hz, 1 H), 3.70 (dd, J=12.0, 4.7 Hz, 1H), 3.62 (s, 1H), 2.45 (s, 1H), 2.31 (d, J = 14.4 Hz, 1H), 2.19 (t, J = 13.6 Hz, 1H), 1.82 (t, J =13.3 Hz, 1H), 1.26 (d, J = 6.4 Hz, 3H). LCMS m/z 461.16 (M+l)⁺
Compound 272	222;	Ή NMR (400 MHz, DMSO-d6) δ 9.33 (d, J = 10.6 Hz, 1H), 8.89 (d, J = 11.1 Hz, 1H), 8.28 (s, 1H), 7.02 (d, J =15.6 Hz, 1H), 5.63 (s, 2H), 4.71 (t, J = 11.4 Hz, 1H),

314

Product	Starting Material and Amine	NMR, LCMS
ο x / \/~~\ ° \ Λ ^ΖΙο \—( '7^=\ ζ-²; X	nh₂A hn^n	4.41 (t, J = 4.4 Hz, 1H), 3.99 (dd, J= 12.0, 4.0 Hz, 1 H), 3.70 (dd,J= 11.9, 4.8 Hz, 1 H), 3.63 (s, 2H), 2.45 (s, 1H), 2.31 (s, 3H), 2.20 (t, J = 13.7 Hz, 1H), 1.83 (t, J = 13.3 Hz, 1H), 1.27 (d, J = 6.4 Hz, 3H). LCMS m/z 436.19 (M+l)⁺
Compound 273 ΟΗ oÔR _NRA N-J Q	222; nh₂cj	Ή NMR (400 MHz, DMSO-d6) δ 9.36 (d, J = 10.2 Hz, 1H), 8.91 (d, J = 11.2 Hz, 1H), 8.29 (s, 1H), 7.04 (s, 1H), 5.52 (s, 1H), 4.69 (t, J = 11.2 Hz, 1H), 4.43 (t, J = 7.1 Hz, 3H), 3.99 (dd, J = 12.0, 4.0 Hz, 1H), 3.73 (d, J = 9.7 Hz, 3H), 3.65 (s, 1H), 3.27 (td, J = 11.0,2.7 Hz, 1H), 3.072.97 (m, 1H), 2.45 (s, 1H), 2.33 (d, J = 14.2 Hz, 1H), 2.21 (t, J = 13.6 Hz, 1H), 1.85 (d, J = 14.0 Hz, 1H), 1.80 (s, 1H), 1.67 (ddq, J = 20.7, 13.9, 6.9 Hz, 2H), 1.53 (s, 1H), 1.41 (s, 2H), 1.28 (d, J = 6.4 Hz, 3H), 1.18 (d, J= 12.3 Hz, 1H). LCMS m/z 453.17 (M+l)⁺
Compound 274 ΟΗ oCQ-^cl N-⁴ Ν-Ν	222; nh₂ n-n « a^.	Ή NMR (400 MHz, DMSO-d6) δ 9.36 (d, J = 10.8 Hz, 1H), 8.89 (d, J = 11.2 Hz, 1H), 8.04 (s, 1H), 7.38 - 7.30 (m, 1H), 7.05 (s, 1H), 5.95 (s, 1H), 4.82 (t, J = 5.9 Hz, 2H), 4.66 (t, J= 11.5 Hz, 1H), 4.46 (dt, J = 17.9,5.3 Hz, 3H), 4.19 (t, J = 5.6 Hz, 1H), 3.98 (dd, J= 11.9, 4.2 Hz, 1 H), 3.68 (td, J = 13.7,

315

Product	Starting Material and Amine	NMR, LCMS
		13.0, 6.5 Hz, 1H), 2.34 (t, J = 12.8 Hz, 2H), 2.27 (s, 1H), 2.15 (t, J = 13.6 Hz, 1H), 1.98 (s, 3H), 1.82 (t, J =13.3 Hz, 1H), 1.27 (d, J = 6.5 Hz, 3H). LCMS m/z 449.20 (M+l)⁺
Compound 275 OH oÔQ*^-01 N 'J H ‘ N-J OH	222; nh₂ OH	’H NMR (400 MHz, DMSO-d6) δ 11.71 (s, 1H), 9.29 (d, J =10.3 Hz, 1H), 8.87 (d, J = 10.9 Hz, 1H), 8.27 (s, 1H), 7.60 (d, J = 2.7 Hz, 1H), 7.41 (dd, J = 9.5, 2.7 Hz, 1H), 7.02 (d, J = 13.0 Hz, 1H), 6.34 (d, J = 9.4 Hz, 1H), 5.38 (s, 2H), 4.68 (t, J =11.2 Hz, 1H), 4.41 (t, J = 4.3 Hz, 1H), 3.98 (dd, J =12.0, 4.0 Hz, 1H), 3.74-3.62 (m, 1H), 2.45 (s, 1H), 2.31 (d, J= 14.6 Hz, lH),2.18(t, J= 13.6 Hz, 1H), 1.82 (t, J =13.3 Hz, 1H), 1.26 (d, J = 6.4 Hz, 3H). LCMS m/z 448.16 (M+l)⁺
Compound 276 OH ₀ÔQ*-^ci Mu N-⁴ Y /N	222; nh₂Y ^NC=/^N	Ή NMR (400 MHz, DMSO-d6) δ 9.34 (d, J = 10.3 Hz, 1H), 8.92 (d, J = 10.7 Hz, 1H), 8.72 (d, J= 1.4 Hz, 1H), 8.69 - 8.60 (m, 2H), 8.39 (s, 1H), 7.04 (s, 1H), 5.88 (s, 2H), 5.52 (s, 1H), 4.72 (t, J= 11.3 Hz, 1H), 4.42 (d, J = 4.5 Hz, 1H), 3.99 (dd, J = 12.0, 4.0 Hz, 1 H), 3.71 (dd, J = 12.0, 4.8 Hz, 1H), 3.64 (s, 1H), 2.47 (s, 1H), 2.32 (d, J = 14.0 Hz, 1H), 2.27-2.16 (m, 1H), 1.89- 1.78 (m, 1H), 1.28 (d, J = 6.4 Hz, 3H).

316

Product	Starting Material and Amine	NMR, LCMS
		LCMS m/z 433.17 (M+l)⁺
Compound 277 OH oÔQ-^ci J, n'J h N-⁴ N=J	222; NH, SN=J	Ή NMR (400 MHz, DMSO-d6) δ 9.27 (d, J = 10.3 Hz, 1H), 8.87 (d, J = 11.1 Hz, 1H), 8.72 (s, 2H), 8.38 (s, 1H), 7.02 (d, J = 13.4 Hz, 1H), 5.72 (s, 2H), 5.50 (s, 1H), 4.69 (t, J =11.2 Hz, 1H), 4.41 (t, J = 4.3 Hz, 1H), 3.98 (dd, J = 12.0, 4.0 Hz, 1H), 3.743.62 (m, 1H), 2.61 (s, 3H), 2.45 (s, 1H), 2.31 (d, J = 14.3 Hz, 1H), 2.26-2.15 (m, 1H), 1.82 (t, J= 13.3 Hz, 1H), 1.26 (d, J = 6.5 Hz, 3H). LCMS m/z 447.17 (M+l)⁺
Compound 278 OH o0Q~^ci N⁴€>°\	222; nh₂ €>°\	*H NMR (400 MHz, DMSO-d6) δ 9.30 (s, 1H), 8.88 (d, J = 10.0 Hz, 1H), 8.32 (s, 1H), 7.29 (dt, J= 10.8, 8.3 Hz, 1H), 7.04 (s, 1H), 6.95 - 6.86 (m, 3H), 5.61 (s, 2H), 5.51 (d, J = 6.1 Hz, 1H), 4.69 (t, J= 10.9 Hz, 1H), 4.41 (s, 1H), 3.98 (dd, J= 12.0, 3.9 Hz, 1H), 3.74 (s, 3H), 3.70 (dd, J= 12.0, 4.8 Hz, 1H), 3.62 (s, 1H), 2.46 (s, 1 H), 2.31 (d, J = 14.2 Hz, 1H), 2.20 (t, J = 13.6 Hz, 1H), 1.82 (t, J = 13.3 Hz, 1H), 1.26 (d, J = 6.5 Hz, 3H). LCMS m/z 461.16 (M+l)⁺
Compound 279	222; nh₂oh	Ή NMR (400 MHz, DMSO-d6) δ 9.61 (s, 1H), 9.30 (d, J= 10.3 Hz, 1H), 8.87 (d, J = 10.9 Hz, 1H), 8.29 (s, 1H), 7.21-7.11 (m, 1H), 7.04 (s, 1H), 6.78 - 6.70

317

Product	Starting Material and Amine	NMR, LCMS
I /—\ o—< x ^ωκΑ O		(m, 3H), 5.55 (s, 2H), 4.69 (s, 1H), 4.41 (t, J = 4.3 Hz, 1H), 3.98 (dd, J = 12.0,3.9 Hz, 1H), 3.70 (dd, J = 12.1, 4.8 Hz, 1H), 3.62 (s, 1H), 2.45 (s, 1H), 2.31 (d, J= 14.4 Hz, 1H), 2.25-2.14 (m, 1H), 1.88 - 1.76 (m, 1H), 1.26 (d, J = 6.4 Hz, 3H). LCMS m/z 447.17 (M+l)⁺
Compound 280 OH oÔ>^cl Λψ' N-J b	222; NH, b	Ή NMR (400 MHz, DMSO-d6) δ 9.36 (d, J = 10.7 Hz, 1H), 9.23 (dd, J = 5.1, 1.7 Hz, 1H), 8.93 (d, J= 10.4 Hz, 1H), 8.43 (s, 1H), 7.77 (dd, J = 8.4, 4.9 Hz, 1H), 7.69 (dd, J = 8.4, 1.7 Hz, 1H), 7.02 (d, J = 14.2 Hz, 1H), 6.02 (s, 2H), 5.55 (s, 1H), 4.73 (t, J = 11.1 Hz, 1H), 4.42 (d, J = 4.6 Hz, 1 H), 3.99 (dd, J = 12.1,4.1 Hz, 1H), 3.71 (dd, J= 12.0, 4.9 Hz, 1H), 3.64 (s, 1H), 2.47 (s, 1H), 2.32 (d, J = 14.1 Hz, 1H), 2.22 (t, J = 13.6 Hz, 1H), 1.84 (t, J = 13.3 Hz, 1H), 1.28 (d, J = 6.4 Hz, 3H). LCMS m/z 433.13 (M+l)⁺
Compound 281 OH oÔl>^ci Ατό KH OH	222; nh₂ OH	Ή NMR (400 MHz, DMSO-d6) δ 9.40 (s, 1H), 8.92 (d, J =11.4 Hz, 1H), 8.26 (d, J= 1.1 Hz, 1H), 7.43 - 7.24 (m, 5H), 7.05 (s, 1H), 5.86 (s, 1H), 5.52 (s, 1H), 4.99 (s, 1H), 4.71 (t, J = 11.2 Hz, 1H), 4.62-4.46 (m, 2H), 4.43 (s, 1H), 4.00 (dd, J = 12.1,4.0 Hz, 1H), 3.72 (dd, J = 12.0, 4.9 Hz, 1H), 3.63 (s, 1H), 2.44 (s, 1H),

318

Product	Starting Material and Amine	NMR, LCMS
		2.33 (d, J= 14.1 Hz, 1H), 2.21 (t, J = 13.2 Hz, 1H), 1.84 (t, J = 13.3 Hz, 1H), 1.29 (d, J = 6.4 Hz, 3H). LCMS m/z 461.16 (M+l)⁺
Compound 282 OH o0Q-^c' ,Ν^ρΑ Α, ^N. J H X	222; NH₂X	Ή NMR (400 MHz, DMSO-d6) δ 9.38 (s, 1H), 8.93 (d, J= 12.0 Hz, 1H), 8.25 (s, 1H), 7.03 (d, J= 14.3 Hz, 1H), 5.52 (s, 1H), 4.70 (t, J = 11.1 Hz, 1H), 4.58 (t, J = 5.0 Hz, 2H), 4.42 (s, 1H), 3.99 (dd, J = 12.1, 3.9 Hz, 1H), 3.773.67 (m, 3H), 3.63 (s, 1H), 3.24 (s, 3H), 2.45 (s, 1H), 2.32 (d, J = 14.2 Hz, 1H), 2.21 (t, J= 13.6 Hz, 1H), 1.84 (t, J = 13.3 Hz, 1H), 1.28 (d, J = 6.5 Hz, 3H). LCMS m/z 399.17 (M+l)⁺
Compound 283 OH oôj JA N'^J A F	222; nh₂A F	’H NMR (400 MHz, DMSO-d6) δ 9.39 (s, 1H), 8.97 (d, J= 10.8 Hz, 1H), 8.33 (s, 1H), 7.04 (s, 1H), 6.66 - 6.32 (m, 1H), 5.52 (s, 1H), 5.04 (td, J = 16.1,3.0 Hz, 2H), 4.75 (t, J = 10.9 Hz, 1H), 4.42 (s, 1H), 4.00 (dd, J = 12.1,4.0 Hz, 1 H), 3.71 (dd, J = 12.1, 4.8 Hz, 1H), 3.64 (s, 1H), 2.46 (s, 1H), 2.33 (d, J= 14.2 Hz, 1H), 2.23 (t, J = 13.6 Hz, 1H), 1.85 (t, J =13.3 Hz, 1H), 1.29 (d, J = 6.5 Hz, 3H). LCMS m/z 405.13 (M+l)⁺
Compound 284	222;	‘H NMR (400 MHz, DMSO-d6) δ 9.37 (s, 1H), 8.91 (d, J= 10.3 Hz, 1H), 8.20 (d, J= 1.7 Hz, 1H), 7.03 (d, J =

319

Product	Starting Material and Amine	NMR, LCMS
τ CL -Z /—\ °—\ » X >-o \__/\__/ X O	nh₂ ho'	13.2 Hz, 1H), 5.53 (s, 1H), 5.07 (s, 1H), 4.70 (t, J = 11.1 Hz, 1H), 4.46 - 4.37 (m, 2H), 4.29 (dd, J = 13.9, 7.1Hz, 1H), 3.99 (dd, J = 12.1,4.0 Hz, 1H), 3.77 - 3.68 (m, 2H), 3.64 (s, 1H), 2.45 (s, 1H), 2.33 (d, J = 14.2 Hz, 1H), 2.22 (td, J = 14.1, 13.5, 4.6 Hz, 1H), 1.84 (t, J= 13.3 Hz, 1H), 1.39-1.21 (m, 4H), 0.91 (t, J = 7.4 Hz, 3H). LCMS m/z 413.17 (M+l)⁺
Compound 285 OH Ù>^CI N-J r	222; nh₂ -b 1	Ή NMR (400 MHz, DMSO-d6) δ 9.25 (d, J = 10.3 Hz, 1H), 8.83 (d, J = 11.1 Hz, 1H), 8.16 (s, 1H), 7.41 (s, 1H), 7.04 (s, 1H), 5.44 (s, 2H), 4.65 (t, J =11.2 Hz, 1H), 4.40 (t, J = 4.2 Hz, 1H), 4.25 (s, 2H), 3.97 (dd, J = 11.9, 3.9 Hz, 1H), 3.70 (s, 3H), 2.44 (s, 1H), 2.30 (d, J= 14.7 Hz, 1H), 2.26 (s, 3H), 2.16 (t, J = 13.6 Hz, 1H), 1.81 (t, J= 13.2 Hz, 1H), 1.26 (d, J = 6.5 Hz, 4H). LCMS m/z 449.20 (M+l)⁺
Compound 286 OH oCjQ>-^ci N-J —=N	222; nh₂Un	’H NMR (400 MHz, DMSO-d6) δ 9.41 (d, J = 10.3 Hz, 1H), 8.96 (d, J = 10.8 Hz, 1H), 8.35 (s, 1H), 7.05 (s, 1H), 5.52 (s, 1H), 4.73 (t, J = 6.3 Hz, 3H), 4.42 (t, J = 4.4 Hz, 1H), 4.00 (dd, J = 12.2, 4.0 Hz, 1H), 3.71 (dd, J = 11.9, 4.9 Hz, 1H), 3.65 (s, 1H), 3.19 (t, J = 6.3 Hz, 2H), 2.45 (s, 1H), 2.33 (d, J= 14.1 Hz, 1H), 2.22 (dd, J =

320

Product	Starting Material and Amine	NMR, LCMS
		14.5, 12.7 Hz, 1H), 1.84 (dd, J = 14.4, 12.2 Hz, 1H), 1.29 (d, J = 6.5 Hz, 3H). LCMS m/z 394.16 (M+l)⁺
Compound 287 OH °0Q“^ci Y’ N-J ^nXq	222; nh₂	^lH NMR (400 MHz, DMSO-d6) δ 9.36 (s, 1H), 8.93 (d, J= 11.0 Hz, 1H), 8.30 (s, 1H), 7.05 (s, 1H), 5.53 (s, 1H), 4.70 (t, J = 11.5 Hz, 1H), 4.52 - 4.39 (m, 3H), 4.11 (p, J = 6.7 Hz, 1H), 3.99 (dd, J = 12.0, 4.0 Hz, 1 H), 3.71 (dd, J = 12.0, 5.0 Hz, 1H), 3.07 (dd, J = 10.1, 5.4 Hz, 1H), 2.87 - 2.77 (m, 1H), 2.44 (s, 1H), 2.35 (dd, J = 16.6, 9.3 Hz, 2H), 2.22 (td, J =13.7, 5.0 Hz, 1H), 2.09 (dd, J =16.6, 6.2 Hz, 1H), 1.84 (t, J = 13.3 Hz, 1H), 1.29 (d, J = 6.5 Hz, 3H), 1.03 (dd, J = 6.8, 4.7 Hz, 6H). LCMS m/z 480.22 (M+l)⁺
Compound 288 OH oÔQ*-^cl/yM-, n--^j h₂n~4 0	222; nh₂ 3 h₂nO	¹H NMR (400 MHz, DMSO-d6) δ 9.35 (d, J = 10.2 Hz, 1H), 8.90 (d, J = 11.0 Hz, 1H), 8.21 (s, 1H), 7.47 (s, 1H), 7.04 (s, 1H), 6.98 (s, 1H), 5.52 (s, 1H), 4.69 (t, J = 11.5 Hz, 1H), 4.58 (t, J = 6.6 Hz, 2H), 4.42 (d, J = 4.5 Hz, 1H), 3.99 (dd, J = 12.0, 4.0 Hz, 1H), 3.71 (dd, J= 12.1, 4.8 Hz, 1H), 3.09 (q, J = 6.8 Hz, lH),2.72(t, J = 6.6 Hz, 2H), 2.49 - 2.35 (m, 1H), 2.32 (d, J = 13.9 Hz, 1H), 2.25-2.13 (m, 1H), 1.89- 1.77 (m, 1H), 1.28 (d, J = 6.5 Hz,

321

Product	Starting Material and Amine	NMR, LCMS
		3H). LCMS m/z 412.17 (M+l)⁺
Compound 289 OH <ÔQ“^ci -i-yV' N-J b	222; NH, 6	’H NMR (400 MHz, DMSO-d6) δ 9.38 (d, J = 10.1 Hz, 1 H), 8.95 (d, J = 10.9 Hz, 1H), 8.21 (s, 1H), 7.04 (s, 1H), 5.52 (s, 1H), 4.79 (s, 1H), 4.73 (s, 2H), 4.42 (t, J = 4.4 Hz, 1H), 4.00 (dd, J = 12.1,4.0 Hz, 1H), 3.79 - 3.67 (m, 3H), 3.54-3.47 (m, 1H), 2.47 (s, 1H), 2.33 (d, J = 14.2 Hz, 1H), 2.29-2.18 (m, 1H), 1.85 (t, J= 13.2 Hz, 2H), 1.75 (d, J = 11.3 Hz, 2H), 1.50 (t, J= 12.3 Hz, 2H), 1.29 (d, J = 6.4 Hz, 3H). LCMS m/z 457.19 (M+l)⁺
Compound 290 OH _QO> yv N-J Sr^N	222; nh₂V» Sr^N	’H NMR (400 MHz, DMSO-d6) δ 9.33 (s, 1H), 8.91 (s, 1H), 8.23 (s, 1H), 7.84 (s, 1H), 7.48 (s, 1H), 7.03 (s, 1H), 5.49 (s, 2H), 4.67 (t, J = 11.1 Hz, 1H), 4.41 (s, 1H), 4.09 (q, J = 7.3 Hz, 2H), 3.98 (dd, J = 11.9, 3.9 Hz, 1H), 3.70 (dd, J = 12.1, 4.8 Hz, 1H), 3.61 (s, 1H), 2.44 (s, 1H), 2.31 (d, J= 14.6 Hz, 1H), 2.20 (dd, J = 14.4, 12.7 Hz, 1H), 1.83 (t, J = 13.3 Hz, 1H), 1.33 (t, J = 7.2 Hz, 3H), 1.25 (t, J = 7.1 Hz, 3H). LCMS m/z 449.15 (M+l)⁺
Compound 291	222; nh₂	Ή NMR (400 MHz, DMSO-d6) δ 9.33 (d, J = 10.1 Hz, 1 H), 8.90 (d, J = 10.7 Hz, 1H), 8.43 (s, 1H), 8.26 (d, J = 6.7 Hz, 2H), 7.04 (s, 1H), 5.61 (s, 2H), 5.50 (s,

322

Product	Starting Material and Amine	NMR, LCMS
OH oÔQ-^cl N-J ΓΚ Ct		1H), 4.70 (t, J = 11.3 Hz, 1H), 4.41 (s, 1H), 3.99 (dd, J =12.1, 3.9 Hz, 1H), 3.70 (dd, J = 12.0, 4.8 Hz, 1H), 3.62 (s, 1H), 2.45 (s, 1H), 2.32 (d, J= 11.4 Hz, 1H), 2.20 (t, J =13.6 Hz, 1H), 1.83 (t, J =13.3 Hz, 1H), 1.27 (d, J = 6.5 Hz, 3H). LCMS m/z 422.11 (M+l)⁺
Compound 292 OH oCjlJ¹ K'ï H ' N-J Y-N // \\	222; nh₂ Y-N // \\	‘H NMR (400 MHz, DMSO-d6) δ 9.35 (d, J = 10.5 Hz, 1H), 8.92 (d, J = 11.0 Hz, 1H), 8.54 (d, J = 4.6 Hz, 1H), 8.35 (s, 1H), 7.85 (td, J = 7.7, 1.9 Hz, 1H), 7.41 -7.33 (m, 2H), 7.04 (s, 1H), 5.77 (s, 2H), 4.72 (t, J = 11.1 Hz, 1H), 4.42 (t, J = 4.2 Hz, 1H), 3.99 (dd, J = 12.1, 4.0 Hz, 1H), 3.71 (dd, J = 12.1, 4.8 Hz, 1H), 2.47 (s, 1H), 2.32 (d, J = 14.0 Hz, 1H), 2.27-2.16 (m, 1H), 1.83 (t, J= 13.3 Hz, 1H), 1.28 (d, J = 6.4 Hz, 3H). LCMS m/z 432.13 (M+l)⁺
Compound 293 OH oOq~^c' <YT^% N⁴ Q-°\ F	222; nh₂ ^F °\	Ή NMR (400 MHz, DMSO-d6) δ 9.32 (d, J = 10.7 Hz, 1H), 8.91 (d, J = 11.0 Hz, 1H), 8.34 (s, 1H), 7.04 (s, 1H), 6.83 (dt, J= 11.2, 2.3 Hz, 1H), 6.78 (d, J = 2.5 Hz, 1H), 6.72 (dd, J = 9.0, 1.9 Hz, 1H), 5.63 (s, 2H), 5.51 (s, 1H), 4.70 (t, J = 11.2 Hz, 1H), 4.41 (s, 1 H), 3.99 (dd, J = 12.1, 4.0 Hz, 1H), 3.76 (s, 3H), 3.70 (dd, J = 12.0, 4.8 Hz, 1H), 3.63 (s, 1H), 2.46 (s, 1H),

323

Product	Starting Material and Amine	NMR, LCMS
		2.31 (d, J = 14.2 Hz, 1H), 2.27-2.16 (m, 1H), 1.83 (t, J= 13.3 Hz, 1H), 1.27 (d, J = 6.5 Hz, 3H). LCMS m/z 479.18 (M+l)⁺
Compound 294 OH oÔl>^ciκ-Υ'Ψ' N J h	222; nh₂ Y-N y	*H NMR (400 MHz, DMSO-d6) δ 9.34 (d, J = 10.4 Hz, 1H), 8.90 (d, J =11.4 Hz, 1H), 8.38 (s, 1H), 8.32 (s, 1H), 7.66 (d, J = 8.2 Hz, 1H), 7.28 (d, J = 7.9 Hz, 1H), 7.04 (s, 1H), 5.71 (s, 2H), 4.71 (t, J = 11.3 Hz, 1H), 4.41 (t, J = 4.4 Hz, 1 H), 3.99 (dd, J= 12.0, 4.0 Hz, 1H), 3.71 (dd, J = 12.1, 4.8 Hz, 1H), 2.46 (s, 1H), 2.29 (s, 3H), 2.20 (t, J =13.6 Hz, 1H), 1.83 (t, J= 13.3 Hz, 1H), 1.27 (d, J = 6.5 Hz, 3H). LCMS m/z 446.17 (M+l)⁺
Compound 295 OH N-J /°	222; nh₂ /°	’H NMR (400 MHz, DMSO-d6) δ 9.30 (d, J = 10.5 Hz, 1H), 8.87 (d, J = 10.7 Hz, 1H), 8.26 (s, 1H), 7.36 - 7.29 (m, 2H), 7.04 (s, 1H), 6.98 6.90 (m, 2H), 5.56 (s, 2H), 4.68 (t, J= 11.2 Hz, 1H), 4.41 (d, J = 4.2 Hz, 1H), 3.98 (dd, J =12.1, 3.9 Hz, 1 H), 3.74 (s, 3H), 3.70 (dd, J= 12.0, 4.7 Hz, 1H), 2.44 (s, 1H), 2.30 (d, J= 14.6 Hz, 1H), 2.18 (t, J = 13.6 Hz, 1H), 1.82 (t, J= 13.3 Hz, 1H), 1.26 (d, J = 6.4 Hz, 3H). LCMS m/z 461.16 (M+l)⁺
Compound 296	222;	Ή NMR (400 MHz, DMSO-d6) δ 9.40 (d, J = 10.2 Hz, 1H), 8.98 (d, J = 11.1 Hz, 1H), 8.54

324

Product _r	Starting Material and Amine	NMR, LCMS
H /—\ °—\ IZ A >-O \__Z\ / i O	nh₂	(d, J = 5.4 Hz, 1H), 8.39 (s, 1H), 7.26 (d, J = 3.9 Hz, 1H), 7.19 (d, J = 5.4 Hz, 1H), 7.04 (s, 1H), 5.79 (s, 2H), 4.73 (t, J = 11.3 Hz, 1H), 4.42 (t, J = 4.5 Hz, 1H), 3.99 (dd, J = 12.0, 4.0 Hz, 1H), 3.80 -3.67(m, 1H), 2.54 (s, 3H), 2.54 - 2.45 (m, 1H), 2.33 (d, J =14.1 Hz, 1H), 2.23 (d, J= 13.8 Hz, 1H), 1.85 (t, J= 13.3 Hz, 1H), 1.28 (d, J = 6.4 Hz, 3H). LCMS m/z 446.17 (M+l)⁺
Compound 297 OH οΟίΑ¹</r N⁴$=N	222; nh₂$=_N ^ογΥ	Ή NMR (400 MHz, DMSO-d6) δ 9.38 (d, J = 10.6 Hz, 1H), 8.96 (d, J = 11.1 Hz, 1H), 8.36 (s, 1H), 5.79 (s, 2H), 4.73 (t, J = 11.4 Hz, 1H), 4.42 (t, J = 4.4 Hz, 1H), 3.99 (dd, J= 12.0, 4.1 Hz, 1 H), 3.71 (dd, J = 12.1, 4.8 Hz, 1H), 3.64 (s, 2H), 2.46 (s, 1H), 2.32 (d, J =13.8 Hz, 1H), 2.27-2.16 (m, 2H), 2.20 (s, 3H), 1.99 (s, 3H), 1.84 (t, J =13.3 Hz, 1H), 1.28 (d, J = 6.4 Hz, 3H). LCMS m/z 450.19 (M+l)⁺
Compound 298 OH oOo-^c\|AV' N-J \-N F	222; nh₂ \-N F	Ή NMR (400 MHz, DMSO-d6) δ 9.40 (s, 1H), 8.97 (s, 1H), 8.50 (d, J = 2.4 Hz, 1H), 8.33 (s, 1H), 8.13-8.04 (m, 1H), 7.04 (s, 1H), 5.84 (d, J = 2.0 Hz, 2H), 5.52 (s, 1H), 4.71 (s, 1H), 4.41 (s, 1H), 3.99 (dd, J = 12.0, 4.0 Hz, 1H), 3.70 (dd, J = 12.0, 4.8 Hz, 1H), 3.63 (s, 1H), 2.45 (s, 1H), 2.31 (d, J= 14.1 Hz, 1H), 2.20 (t, J = 13.6

325

Product	Starting Material and Amine	NMR, LCMS
		Hz, 1H), 1.83 (t, J= 13.3 Hz, 1H), 1.28 (d, J = 6.5 Hz, 3H). LCMS m/z 468.16 (M+l)⁺
Compound 299 OH <Ô0~^ci N-J III·· ho'	222; nh₂ un· HO	Ή NMR (400 MHz, DMSO-d6) δ 9.36 (d, J = 10.3 Hz, 1H), 8.91 (d, J = 10.8 Hz, 1H), 8.28 (s, 1H), 7.02 (d, J =15.6 Hz, 1H), 5.53 (s, 1H), 4.68 (t, J = 11.3 Hz, 1H), 4.59 (s, 1H), 4.44 (dd, J = 10.8, 7.0 Hz, 3H), 3.99 (dd, J= 12.0, 4.0 Hz, 1H), 3.71 (dd, J =12.0, 4.9 Hz, 1H), 3.64 (s, 1H), 3.25 (q, J = 5.9, 5.4 Hz, 1H), 2.45 (s, 1H), 2.33 (d, J = 14.0 Hz, 1H), 2.27-2.15 (m, 1H), 1.93 (dq, J= 13.7, 7.4 Hz, 1H), 1.83 (dd, J = 14.5, 12.2 Hz, 1H), 1.58 (dq, J = 14.2, 7.4 Hz, 1H), 1.47 (dt, J= 13.1, 6.4 Hz, 1H), 1.28 (d, J = 6.5 Hz, 3H), 0.86 (dd, J = 11.0, 6.7 Hz, 3H). LCMS m/z 427.21 (M+l)⁺
Compound 300 OH <Oq~^ci N-J ^•OH OH	222; nh₂^••OH OH	Ή NMR (400 MHz, DMSO-d6) δ 7.87 (s, 1H), 6.96 (s, 1H), 5.40 (s, 1H), 5.13 (s, 1H), 4.86 (s, 1H), 4.44 (dd, J = 13.8, 3.5 Hz, 1H), 4.39 (s, 1H), 4.22 - 4.08 (m, 2H), 3.93 (dd, J= 11.8, 4.2 Hz, 1H), 3.78 (s, 1 H), 3.64 (dd, J= 11.8, 5.3 Hz, 1H), 3.29 (s, 1H), 3.10 (s, 1H), 2.24 (s, 1H), 2.19 (d, J= 13.5 Hz, 1H), 2.05 (d, J = 13.3 Hz, 1H), 1.58 (dd, J = 13.3, 11.7 Hz, 1H), 1.30-1.16 (m, 1H), 1.03 (d, J = 6.2 Hz, 3H).

326

Product	Starting Material and Amine	NMR, LCMS
		LCMS m/z 415.15 (M+l)⁺
Compound 301 OH oôi>^cl IA 0^	222; NH, A	Ή NMR (400 MHz, DMSO-d6) δ 8.02 (s, 1H), 7.24 - 7.07 (m, 3H), 6.96 (s, 1H), 6.95 (s, 1H), 5.41 (s, 1H), 4.74 (t, J = 5.1 Hz, 2H), 4.46 (t, J = 5.1 Hz, 2H), 4.39 (t, J = 4.6 Hz, 1H), 4.12 (d, J= 11.4 Hz, 1H), 3.93 (dd, J= 11.9, 4.2 Hz, 1H), 3.64 (dd, J = 11.8,5.2 Hz, 1H), 3.10 (s, 1H), 2.26 (s, 1H), 2.17 (d, J =13.5 Hz, 1H), 2.05 (d, J= 13.3 Hz, 1H), 1.56 (dd, J = 13.4, 11.7 Hz, 1H), 1.25 (dd, J= 13.3, 11.2 Hz, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 479.18 (M+l)⁺
Compound 302 L F OA N —' OH	223; nh₂ OH	’H NMR (400 MHz, DMSO-d6) δ 9.55 (s, 1H), 7.94 (s, 1H), 7.34 (s, 1H), 7.21 -7.13 (m, 2H), 6.77 - 6.70 (m, 2H), 5.39 (s, 2H), 4.21 (t, J = 10.6 Hz, 2H), 4.10 (dd, J = 11.6, 2.5 Hz, 1H), 3.04 (s, 1H), 2.32 (d, J= 13.5 Hz, 1H), 2.10 (d, J = 13.5 Hz, 1H), 1.66 (dd, J = 13.5, 11.7 Hz, 1H), 1.36-1.21 (m, 1H), 1.01 (d, J = 6.2 Hz, 3H). LCMS m/z 467.13 (M+l)⁺
Compound 303	223; nh₂	Ή NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.35 (s, 1H), 7.15 (td, J = 8.0, 1.3 Hz, 1H), 6.74 (dd, J = 10.2,6.3 Hz, 3H), 5.55 (d, J = 5.5 Hz, 1H), 4.53 (dd, J = 13.7,3.9 Hz, 1H), 4.444.34 (m, 1H), 4.27-4.17

327

Product	Starting Material and Amine	NMR, LCMS
ο M— O>—< z° ^zJi^ JL Z '^ O X		(m, 3H), 4.17-4.10 (m, 1H), 3.88 (d, J = 5.4 Hz, 2H), 3.07 (s, 1H), 2.30 (s, 1H), 2.26 (s, 3H), 2.13 (d, J = 13.6 Hz, 1H), 1.69 (t, J =11.8 Hz, 1H), 1.32 (t, J =12.4 Hz, 1H), 1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 525.23 (M+l)⁺
Compound 304 F. ^F °O>^ci M'TN-J HO^P	223; nh₂HO^P	Ή NMR (400 MHz, DMSO-d6) δ 7.96 (d, J = 7.3 Hz, 1H), 7.48 (d, J = 7.3 Hz, 1H), 7.36 (s, 1H), 7.26 - 7.13 (m, 3H), 5.65 (t, J = 3.8 Hz, 1H), 5.11 (s, 1H), 4.44 (d, J = 13.5 Hz, 1H), 4.38-4.11 (m,4H), 3.08 (s, 1H), 2.32 (d, J = 4.9 Hz, 3H), 2.14 (d, J = 13.5 Hz, 1H), 1.76- 1.64 (m, 1H), 1.33 (s, 1H), 1.04 (d, J = 6.3 Hz, 3H). LCMS m/z 495.16 (M+l)⁺
Compound 305 F F oUL>^CIm « N J N—'	223; nh₂M N—'	’H NMR (400 MHz, DMSO-d6) δ 9.11 (d, J = 1.5 Hz, 1H), 8.68 (d, J = 5.2 Hz, 1H), 7.99 (s, 1H), 7.40 (dd, J = 5.3, 1.5 Hz, 1H), 7.35 (s, 1H), 4.76 (t, J = 7.1 Hz, 2H), 4.21 (t, J = 10.7 Hz, 2H), 4.10 (dd, J= 11.7, 2.5 Hz, 1H), 3.34 (s, 2H), 3.09-2.98 (m, 1H), 2.33 -2.25 (m, 1H), 2.12 (d, J = 13.7 Hz, 1H), 1.66 (dd, J= 13.6, 11.6 Hz, 1H), 1.32 (dd, J = 13.7, 11.2 Hz, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 467.17 (M+l)⁺
Compound 306	223;	'H NMR (400 MHz, DMSO-d6) δ 8.01 (s, 1H), 7.36 (s, 1H), 4.53 (t, J = 5.3 Hz, 1H), 4.35

328

Product	Starting Material and Amine	NMR, LCMS
τ CL xV X'x /-χρ-λ^ iz X X \__/\__/ -n \·ζ o	nh₂4 HO	(td, J = 7.9, 7.3, 2.7 Hz, 2H), 4.23 (t, J = 10.6 Hz, 2H), 4.16 (d, J =11.3 Hz, 1H), 3.25 (td, J = 5.6, 1.6 Hz, 2H), 3.09 (s, 1H), 2.35 (d, J = 14.0 Hz, lH),2.14(d, J = 13.6 Hz, 1H), 1.91 (dq, J = 13.5, 7.5 Hz, 1H), 1.72 (t, J= 12.7 Hz, 1H), 1.62 -1.30(m, 3H), 1.05 (d, J = 6.2 Hz, 3H), 0.86 (d, J = 6.7 Hz, 3H). LCMS m/z 447.12 (M+l)⁺
Compound 307 F. F oO> ΛΥ' N-J V	223; nh₂ Ot° ^so	‘H NMR (400 MHz, DMSO-d6) δ 8.03 (s, 1H), 7.36 (s, 1H), 4.49 (d, J = 6.4 Hz, 2H), 4.23 (t, J =10.7 Hz, 2H), 4.18 -4.11 (m, 1H), 3.272.84 (m, 6H), 2.35 (d, J = 13.8 Hz, 1H), 2.14 (d, J = 13.5 Hz, 2H), 1.83 (q, J = 12.2, 10.7 Hz, 1H), 1.71 (t, J = 12.8 Hz, 1H), 1.39- 1.28 (m, 1H), 1.04 (d, J = 6.2 Hz, 3H). LCMS m/z 493.13 (M+l)⁺
Compound 308 F F °OA^ci ^N,J H N-J /°	223; nh₂ d/°	‘H NMR (400 MHz, DMSO-d6) δ 7.98 (s, 1H), 7.34 (s, 1H), 7.31 7.19 (m, 2H), 7.05 - 6.91 (m, 2H), 5.45 (s, 2H), 4.56 (td, J = 8.5, 4.2 Hz, 1H), 4.21 (t, J= 10.6 Hz, 2H), 4.11 (dd, J= 11.6, 2.5 Hz, 1H), 3.83 (dt, J = 11.7, 4.4 Hz, 2H), 3.503.43 (m, 2H), 3.02 (d, J = 19.3 Hz, 1H), 2.32 (d, J = 13.5 Hz, 1H), 2.10 (d, J= 13.5 Hz, 1H), 1.94 (dd, J = 13.1,3.8 Hz, 2H), 1.66 (dd, J = 13.5, 11.7 Hz, 1H), 1.54 (dtd, J = 12.9, 9.0, 4.0 Hz, 2H), 1.42- 1.23 (m,

329

Product	Starting Material and Amine	NMR, LCMS
		1H), 1.01 (d, J = 6.2 Hz, 3H). LCMS m/z 551.20 (M+l)⁺
Compound 309 F. F ô>^clκλΛ N-J	223; nh₂	’H NMR (400 MHz, DMSO-d6) δ 7.97 (s, 1H), 7.35 (s, 1H), 4.27 4.18 (m, 4H), 4.18-4.10 (m, 1H), 3.87 - 3.78 (m, 2H), 3.24 (td, J = 11.7, 2.2 Hz, 2H), 3.08 (s, 1H), 2.34 (d, J= 13.4 Hz, 1H), 2.13 (d, J = 13.6 Hz, 1H), 2.04 (ddd, J =11.5, 7.6, 4.0 Hz, 1H), 1.76- 1.65 (m, 1H), 1.39 (d, J = 10.8 Hz, 2H), 1.36- 1.25 (m, 1H), 1.24 (d, J = 4.6 Hz, 1H), 1.20 (dd, J = 12.1,4.4 Hz, 1H), 1.04 (d, J = 6.2 Hz, 3H). LCMS m/z 459.18 (M+l)⁺
Compound 310 F. F OO^-01 yN Μ Ά_ζ ^N.J H N-⁴R?	223; nh₂A	Ή NMR (400 MHz, DMSO-d6) δ 7.89 (s, 1H), 7.34 (s, 1H), 6.86 (dd, J = 8.2, 1.7 Hz, 1H), 6.80 (t, J = 7.8 Hz, 1H), 6.65 (dd, J = 7.6, 1.7 Hz, 1H), 5.47 (s, 2H), 4.32 (dd, J = 5.8, 2.7 Hz, 2H), 4.26 (dd, J = 5.6, 2.9 Hz, 2H), 4.21 (t, J = 10.5 Hz, 2H), 4.13 (dd, J= 11.6, 2.5 Hz, 1H), 3.06 (d, J = 10.0 Hz, 1H), 2.33 (d, J = 13.6 Hz, 1H), 2.11 (d, J = 13.6 Hz, 1H), 1.70 (dd, J= 13.6, 11.6 Hz, 1H), 1.32 (dd, J= 13.6, 11.3 Hz, 1H), 1.02 (d, J = 6.3 Hz, 3H). LCMS m/z 509.16 (M+l)⁺
Compound 311	223;	Ή NMR (400 MHz, DMSO-d6) δ 8.02 (s, 1H), 7.35 (s, 1H), 4.32 (d, J = 7.5 Hz, 2H), 4.22 (t, J= 10.5 Hz, 2H), 4.13 (dd, J = 11.7, 2.5 Hz,

330

Product	Starting Material and Amine	NMR, LCMS
ο Jw cU- / \/~\ xK X ²¹“ \_j\—/ ixJO⁵Z '----x/	N H, b	1H), 3.76 (td, J = 8.0, 5.5 Hz, 1H), 3.69 - 3.58 (m, 2H), 3.48 - 3.42 (m, 1H), 3.07 (t, J = 8.1 Hz, 1H), 2.71 (p, J = 6.7 Hz, 1H), 2.34 (dt, J= 13.9, 2.5 Hz, 1 H), 2.13 (dd, J = 13.8, 2.7 Hz, 1H), 1.96 -1.86 (m, 1H), 1.70 (dd, J = 13.6, 11.6 Hz, 1H), 1.59 (dt, J = 13.1, 6.6 Hz, 1H), 1.33 (dd, J = 13.6, 11.3 Hz, 1H), 1.04 (d, J = 6.3 Hz, 3H). LCMS m/z 445.18 (M+l)⁺
Compound 312 F. ^F OA' N-⁴	223; nh₂ A-	*H NMR (400 MHz, DMSO-d6) δ 7.94 (s, 1H), 7.35 (s, 1H), 4.27 (d, J = 2.5 Hz, 2H), 4.23 (t, J = 10.5 Hz, 2H), 4.14 (dd, J = 11.7, 2.6 Hz, 1H), 3.22 (s, 3H), 3.10 3.03 (m, 1H), 3.01 (s, 2H), 2.39 - 2.30 (m, 1H), 2.17-2.08 (m, 1H), 1.71 (dd, J= 13.5, 11.7 Hz, 1H), 1.34 (dd, J= 13.6, 11.2 Hz, 1H), 1.04 (d, J = 6.3 Hz, 3H), 0.70 (q, J = 4.0 Hz, 2H), 0.51 (q, J = 4.1 Hz, 2H). LCMS m/z 495.18 (M+l)⁺
Compound 313 F F mA^ci N-J DH	223; nh₂ DH	*H NMR (400 MHz, DMSO-d6) δ 8.01 (s, 1H), 7.36 - 7.28 (m, 3H), 7.26 (d, J = 7.6 Hz, 1H), 7.19 (d, J = 7.4 Hz, 1H), 5.53 (s, 2H), 5.23 (t, J = 5.7 Hz, 1H), 4.48 (d, J = 5.4 Hz, 2H), 4.21 (t, J = 10.6 Hz, 2H), 4.16-4.09 (m, 1H), 3.09-2.97 (m, 1H), 2.33 (d, J= 13.5 Hz, 1H), 2.11 (d, J = 13.5 Hz, 1H), 1.73 -1.62 (m, 1H), 1.37- 1.26 (m, 1H), 1.01 (d, J = 6.3 Hz,

331

Product	Starting Material and Amine	NMR, LCMS
		3H). LCMS m/z 481.17 (M+l)⁺
Compound 314 F. F <ÔC*~^ciO N⁴ ^-Q	223; nh₂	’H NMR (400 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.35 (s, 1H), 4.33 (t, J = 7.3 Hz, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.13 (dd, J = 11.7, 2.6 Hz, 1 H), 3.72 (dd, J =11.4, 4.0 Hz, 2H), 3.26 (td, J = 11.1, 2.9 Hz, 2H), 3.082.95 (m, 2H), 2.33 (d, J = 13.5 Hz, 1H), 2.12 (dd, J= 13.7, 2.8 Hz, 1H), 1.81 (d, J = 12.8 Hz, 1H), 1.75- 1.49 (m, 3H), 1.42 (s, 1H), 1.471.36 (m, 1H), 1.39-1.27 (m, 1H), 1.26 - 1.09 (m, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 473.22 (M+l)⁺
Compound 315 F. F °00“^cin^A hH < “P	223; nh₂A	Ή NMR (400 MHz, DMSO-d6) δ 7.71 (s, 1H), 7.38-7.31 (m, 2H), 5.94 (s, 1H), 4.71 (t, J = 6.0 Hz, 2H), 4.44 (t, J = 5.9 Hz, 2H), 4.22 (t, J = 10.7 Hz, 2H), 4.10 (dd, J = 11.7, 2.5 Hz, 1H), 3.09 - 2.97 (m, 1H), 2.25 (d, J = 14.0 Hz, 1H), 2.172.09 (m, 1H), 1.92 (s, 3H), 1.63 (dd, J= 13.5, 11.6 Hz, 1H), 1.36-1.21 (m, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 469.20 (M+l)⁺
Compound 316	223; nh₂b OH	Ή NMR (400 MHz, DMSO-d6) δ 11.64 (s, 1H), 7.99 (s, 1H), 7.57 7.48 (m, 1H), 7.41 (dd, J = 9.5, 2.6 Hz, 1H), 7.34 (s, 1H), 6.32 (d, J = 9.4 Hz, 1H), 5.28 (s, 2H), 4.21 (t, J = 10.6 Hz, 2H), 4.11 (d, J= 10.9 Hz, 1H), 3.05 (s, 1H), 2.33

332

Product	Starting Material and Amine	NMR, LCMS
F. F <00-^ci ΊΑ'^,/z/ N 'J H N-'' OH		(d, J= 13.5 Hz, 1H), 2.11 (d, J= 13.9 Hz, 1H), 1.72-1.61 (m, 1H), 1.36-1.21 (m, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 468.16 (M+l)⁺
Compound 317 F. F OR <yV' N-J Ν^Λΐ	223; nh₂	‘H NMR (400 MHz, DMSO-d6) δ 8.70 - 8.61 (m, 3H), 8.10 (s, 1H), 7.35 (s, 1H), 5.77 (s, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.14 (dd, J = 11.6, 2.5 Hz, 1H), 3.06 (s, 1H), 2.39-2.31 (m, 1H), 2.12 (d, J = 13.6 Hz, 1H), 1.75- 1.64 (m, 1H), 1.38 - 1.27 (m, 1H), 1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 453.17 (M+l)⁺
Compound 318 E F oOl>^ci <y^kA' R ^ΝΛ	223; nh₂H N—(	‘H NMR (400 MHz, DMSO-d6) δ 8.71 (s, 2H), 8.11 (s, 1H), 7.35 (s, 1H), 5.61 (s, 2H), 4.21 (t, J= 10.6 Hz, 2H), 4.12 (dd, J= 11.7, 2.5 Hz, 1H), 3.10-2.98 (m, 1H), 2.61 (s, 3H), 2.33 (d, J= 13.5 Hz, 1H), 2.11 (d, J = 13.4 Hz, 1H), 1.67 (dd, J =13.5, 11.6 Hz, 1H), 1.30 (dd, J = 13.6, 11.2 Hz, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 467.17 (M+l)⁺
Compound 319	223;	Ή NMR (400 MHz, DMSO-d6) δ 8.02 (s, 1H), 7.34 (s, 1H), 7.29 (td, J = 7.3, 1.9 Hz, 1H), 6.94-6.83 (m, 3H), 5.51 (s, 2H), 4.21 (t, J= 10.6 Hz, 2H), 4.12 (dd, J =

333

Product	Starting Material and Amine	NMR, LCMS
F. F oOQ“^ci N 'J ij N'-^J O-°\	nh₂ O-°x	11.6, 2.5 Hz, 1H), 3.74 (s, 3H), 3.10-2.98 (m, 1H), 2.38 -2.29 (m, 1H), 2.15-2.06 (m, 1H), 1.68 (dd, J= 13.5, 11.6 Hz, 1H), 1.32 (dd, J = 13.6, 11.3 Hz, 1H), 1.02 (d, J = 6.3 Hz, 3H). LCMS m/z 481.21 (M+l)⁺
Compound 320 E ^F oO>^CI KA\|iA k j H N-'' OH	223; nh₂ OH	’H NMR (400 MHz, DMSO-d6) δ 9.60 (s, 1H), 7.99 (s, 1H), 7.34 (s, 1H), 7.15 (t, J = 7.8 Hz, 1H), 6.75 - 6.64 (m, 3H), 5.45 (s, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.12 (dd, J = 11.6, 2.5 Hz, 1H), 3.05 (s, 1H), 2.34 (d, J= 13.6 Hz, 1H), 2.15-2.07 (m, 1H), 1.68 (dd, J= 13.5, 11.6 Hz, 1H), 1.31 (dd, J= 13.5, 11.3 Hz, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 467.17 (M+l)⁺
Compound 321 E F oôj JA b	223; nh₂ &	^lH NMR (400 MHz, DMSO-d6) δ 9.22 (dd, J = 5.0, 1.7 Hz, 1H), 8.13 (s, 1H), 7.74 (dd, J = 8.5, 5.0 Hz, 1H), 7.59 (dd, J = 8.5, 1.7 Hz, 1H), 7.35 (s, 1H), 5.91 (s, 2H), 4.22 (t, J =10.6 Hz, 2H), 4.15 (dd, J= 11.8, 2.6 Hz, 1H), 3.07 (s, 1H), 2.35 (d, J= 13.9 Hz, 1H), 2.12 (d, J = 13.5 Hz, 1H), 1.71 (dd, J = 13.6, 11.6 Hz, 1H), 1.33 (dd, J= 13.6, 11.2 Hz, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 453.13 (M+l)⁺
Compound 322	223;	Ή NMR (400 MHz, DMSO-d6) δ 7.95 (d, J = 4.9 Hz, 1H), 7.42 7.32 (m, 5H), 7.32 - 7.24

334

Product	Starting Material and Amine	NMR, LCMS
ο < u. / \ / \ X X ζι Ô ^zc~^z ζ X	nh₂	(m, 1H), 5.78 (d, J = 4.7 Hz, 1H), 4.97 (d, J = 8.1 Hz, 1H), 4.47 (dd, J = 13.9, 4.0 Hz, 1H), 4.40 (dd, J= 13.8, 8.5 Hz, 1H), 4.23 (t, J = 10.6 Hz, 2H), 4.14 (dd, J= 11.7, 2.5 Hz, 1H), 3.07 (t, J = 8.0 Hz, 1H), 2.54 (s, 1H), 2.36 - 2.27 (m, 1H), 2.14 (d, J= 13.6 Hz, 1H), 1.69 (ddd, J= 13.5, 11.6, 5.6 Hz, 1H), 1.33 (dd, J= 13.8, 11.0 Hz, 1H), 1.04 (d, J = 6.3 Hz, 3H). LCMS m/z 481.21 (M+l)⁺
Compound 323 Ε ^F O>^ci K'Y H ’ hH °\	223; nh₂ °\	‘H NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.35 (s, 1H), 4.48 (t, J = 5.2 Hz, 2H), 4.23 (t, J = 10.6 Hz, 2H), 4.13 (dd, J =11.8, 2.5 Hz, 1H), 3.71 (t, J = 5.1 Hz, 2H), 3.24 (s, 3H), 3.07 (t, J = 5.3 Hz, 1H), 2.54 (s, 1H), 2.38 -2.30 (m, lH),2.12(d, J= 13.8 Hz, 1H), 1.70 (dd, J = 13.5, 11.6 Hz, 1H), 1.33 (dd, J= 13.6, 11.3 Hz, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 419.17 (M+l)⁺
Compound 324 E F oOC*^-01 N-⁴ F F	223; nh₂ F	‘H NMR (400 MHz, DMSO-d6) δ 8.03 (s, 1H), 7.35 (s, 1H), 6.62 6.27 (m, 1H), 4.91 (td, J = 15.7,3.3 Hz, 2H), 4.23 (t, J= 10.6 Hz, 2H), 4.16 (dd, J = 11.7, 2.6 Hz, 1H), 3.06 (d, J =10.1 Hz, 1H), 2.54 (s, 1H), 2.35 (d, J= 13.9 Hz, 1H), 2.13 (d, J= 13.5 Hz, 1H), 1.70 (dd, J = 13.5, 11.6 Hz, 1H), 1.33 (dd, J =13.5, 11.3 Hz,

335

Product	Starting Material and Amine	NMR, LCMS
		1H), 1.04 (d, J = 6.2 Hz, 3H). LCMS m/z 425.13 (M+l)⁺
Compound 325 F. F OA^C\| N·^ HC)	223; nh₂H(j/	Ή NMR (400 MHz, DMSO-d6) δ 7.91 (s, 1H), 7.35 (s, 1H), 5.00 (dd, J = 5.7, 1.9 Hz, 1H), 4.31 (dd, J= 13.8, 4.2 Hz, 1H), 4.22 (t, J = 10.3 Hz, 3H), 4.18-4.08 (m, 1H), 3.72 (s, 1H), 3.07 (s, 1H), 2.40 - 2.25 (m, 2H), 2.13 (d, J = 13.5 Hz, 1H), 1.70 (t, J= 12.6 Hz, 1H), 1.32 (dtq, J = 26.5, 13.4, 6.3 Hz, 3H), 1.03 (d, J = 6.3 Hz, 3H), 0.89 (t, J = 7.4 Hz, 3H). LCMS m/z 433.17 (M+l)⁺
Compound 326 F. F °CQ“^ci N-J 1	223; nh₂	Ή NMR (400 MHz, DMSO-d6) δ 7.86 (s, 1H), 7.40 (s, 1H), 7.34 (s, 1H), 5.34 (s, 2H), 4.21 (t, J= 10.6 Hz, 2H), 4.09 (dd, J = 11.7,2.5 Hz, 1H), 3.69 (s, 3H), 3.10-2.99 (m, 1H), 2.31 (d, J = 13.7 Hz, 2H), 2.25 (s, 3H), 2.10 (d, J = 13.4 Hz, 1H), 1.65 (dd, J = 13.6, 11.6 Hz, 1H), 1.30 (dd, J = 13.6, 11.2 Hz, 1H), 1.01 (d, J = 6.3 Hz, 3H). LCMS m/z 469.20 (M+l)⁺
Compound 327	223; nh₂	Ή NMR (400 MHz, DMSO-d6) δ 8.03 (s, 1H), 7.35 (s, 1H), 4.37 (d, J = 7.3 Hz, 2H), 4.23 (t, J= 10.6 Hz, 2H), 4.12 (td, J= 13.5, 13.0, 4.7 Hz, 2H), 3.30 (s, 2H), 3.14-3.00 (m, 2H), 2.88 - 2.72 (m, 1H), 2.42 2.26 (m, 2H), 2.22 - 2.06 (m, 2H), 1.70 (t, J= 12.6 Hz, 1H), 1.33 (dd, J =

336

Product	Starting Material and Amine	NMR, LCMS
\ -Z ' ¹² X O		13.6, 11.3 Hz, 1H), 1.07 - 0.99 (m, 9H). LCMS m/z 500.22 (M+l)⁺
Compound 328 F. F OT^ci <yV' té H₂léZ 0	223; nh₂ H₂léK 0	Ή NMR (400 MHz, DMSO-d6) δ 7.90 (s, 1H), 7.44 (s, 1H), 7.35 (s, 1H), 7.01 - 6.89 (m, 1H), 4.49 (t, J = 6.8 Hz, 2H), 4.22 (t, J= 10.6 Hz, 2H), 4.12 (dd, J= 11.6, 2.5 Hz, 1H), 3.05 (d, J = 13.6 Hz, 1H), 2.67 (t, J = 6.8 Hz, 2H), 2.32 (d, J = 13.7 Hz, 2H),2.12(d, J = 13.7 Hz, 1H), 1.68 (dd, J= 13.5, 11.7 Hz, 1H), 1.32 (dd, J = 13.6, 11.3 Hz, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 432.13 (M+l)⁺
Compound 329 E F °OQ~^ci -Υιη lé ^F /—\ '—o⁷	223; nh₂ ^f“7—\ '—o⁷	Ή NMR (400 MHz, DMSO-d6) δ 7.91 (s, 1H), 7.35 (s, 1H), 4.65 (d, J = 21.8 Hz, 2H), 4.23 (t, J = 10.6 Hz, 2H), 4.15 (dd, J= 11.6, 2.6 Hz, 1 H), 3.74 (ddd, J = 11.5, 5.0,2.5 Hz, 2H), 3.49 (t, J = 10.9 Hz, 3 H), 3.07 (s, 1H), 2.40-2.30 (m, 1H), 2.12 (d, J = 13.6 Hz, 1H), 1.90-1.63 (m, 3H), 1.49 (t, J =12.3 Hz, 2H), 1.34 (dd, J = 13.6, 11.3 Hz, 1H), 1.04 (d, J = 6.2 Hz, 3H). LCMS m/z 477.19 (M+l)⁺

337

Product	Starting Material and Amine	NMR, LCMS
Compound 330 E F oCQ-^ci N-yv' N-J μ \'^N	223; NH, Sl'^N	‘H NMR (400 MHz, DMSO-d6) δ 7.94 (s, 1H), 7.82 (s, 1H), 7.48 (s, 1H), 7.34 (s, 1H), 5.38 (s, 2H), 4.21 (t, J = 10.6 Hz, 2H), 4.09 (q, J = 7.4 Hz, 3H), 3.04 (d, J = 9.9 Hz, 1H), 2.32 (d, J = 13.7 Hz, 2H), 2.11 (d, J = 13.7 Hz, 1H), 1.67 (dd, J = 13.6, 11.6 Hz, 1H), 1.42-1.21 (m, 4H), 1.01 (d, J = 6.2 Hz, 3H). LCMS m/z 469.20 (M+l)⁺
Compound 331 F F ₀O>^ci<yV' N-J FK (T	223; nh₂V	‘H NMR (400 MHz, DMSO-d6) δ 8.41 (s, 1H), 8.21 (s, 1H), 7.96 (s, 1H), 7.34 (s, 1H), 5.50 (s, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.12 (dd, J = 11.7, 2.5 Hz, 1H), 3.12 -3.01 (m, 1H), 2.41 2.27 (m, 1H), 2.11 (d, J = 13.6 Hz, 1H), 1.68 (dd, J = 13.5, 11.6 Hz, 1H), 1.32 (dd, J= 13.6, 11.3 Hz, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 442.16 (M+l)⁺
Compound 332 F. F Oy^ci <yÇ· N-* b	223; nh₂ \-N // \\	Ή NMR (400 MHz, DMSO-d6) δ 8.55 (d, J = 4.1 Hz, 1H), 8.05 (s, 1H), 7.82 (td, J = 7.7, 1.8 Hz, 1H), 7.35 (d, J = 4.7 Hz, 2H), 7.26 (d, J = 7.8 Hz, 1H), 5.66 (s, 2H), 4.22 (t, J= 10.6 Hz, 2H), 4.14 (dd, J = 11.6, 2.5 Hz, 1H), 3.12-3.04 (m, 1H), 2.35 (d, J = 13.9 Hz, 2H), 2.12 (d, J = 13.7 Hz, 1H), 1.70 (dd, J= 13.5, 11.7 Hz, 1H), 1.33 (dd, J= 13.6, 11.2 Hz, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 452.18 (M+l)⁺

338

Product	Starting Material and Amine	NMR, LCMS
Compound 333 F ^F oOA' ΚΑκΑ, Ό H N-J F °\	223; nh₂^-F °\	^XH NMR (400 MHz, DMSO-d6) δ 8.06 (s, 1H), 7.34 (s, 1H), 6.81 (dt, J = 11.1, 2.3 Hz, 1H), 6.76 (t, J =1.9 Hz, 1H), 6.72 - 6.62 (m, 1H), 5.52 (s, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.16-4.07 (m, 1H), 3.76 (s, 3H), 3.05 (t, J = 9.1 Hz, 1H), 2.34 (d, J= 13.9 Hz, 2H), 2.11 (d, J = 13.6 Hz, 1H), 1.68 (dd, J = 13.6, 11.7 Hz, 1H), 1.32 (dd, J= 13.5, 11.3 Hz, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 499.18 (M+l)⁺
Compound 334 F. F oQQ*-^ci N-J Y-N // \\	223; nh₂ Y-N // \\	Ή NMR (400 MHz, DMSO-d6) δ 8.38 (d, J = 2.2 Hz, 1H), 8.01 (s, 1H), 7.63 (dd, J = 8.4, 2.3 Hz, 1H), 7.35 (s, 1H), 7.19 (d, J = 7.9 Hz, 1H), 5.60 (s, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.16 - 3.97 (m, 1H), 3.06 (s, lH),2.34(d, J= 14.2 Hz, 2H), 2.28 (s, 3H), 2.11 (d, J = 13.5 Hz, 1H), 1.69 (dd, J =13.5, 11.7 Hz, 1H), 1.44-1.29 (m, 1H), 1.02 (d, J = 6.3 Hz, 3H). LCMS m/z 466.22 (M+l)⁺
Compound 335	223; nh₂ 0 /	Ή NMR (400 MHz, DMSO-d6) δ 7.97 (s, 1H), 7.34 (s, 1H), 7.33 7.24 (m, 2H), 7.03 - 6.87 (m, 2H), 5.46 (s, 2H), 4.21 (t, J= 10.6 Hz, 2H), 4.15-4.04 (m, 1 H), 3.73 (s, 3H), 3.04 (s, 1H), 2.32 (d, J = 13.9 Hz, 2H), 2.10 (d, J= 13.5 Hz, 1H), 1.66 (dd, J = 13.6, 11.7 Hz, 1H), 1.30 (dd, J = 13.6, 11.3 Hz,

339

Product	Starting Material and Amine	NMR, LCMS
___________ ,Z JÛX^ZA/^Z ΙΖ A /A '\ f Ή *' ωχ) Ο		1H), 1.01 (d, J = 6.3 Hz, 3H). LCMS m/z 481.17 (M+l)⁺
Compound 336 E F oÔl>^ci ΛΎ^% N-J	223; nh₂	Ή NMR (400 MHz, DMSO-d6) δ 8.42 (d, J = 5.1 Hz, 1H), 8.08 (s, 1H), 7.35 (s, 1H), 7.10 (s, 1H), 7.00 (d, J = 5.1 Hz, 1H), 5.58 (s, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.19-4.04 (m, 1 H), 3.06 (s, 1H), 2.44 (s, 3H), 2.35 (d, J = 13.8 Hz, 2H), 2.12 (d, J = 13.5 Hz, 1H), 1.69 (dd, J = 13.5, 11.6 Hz, 1H), 1.32 (dd, J= 13.5, 11.3 Hz, 1H), 1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 466.18 (M+l)⁺
Compound 337 F. F °OQ^ci N-J $=N °Xj^\	223; nh₂S=N ⁰Xj^s.	Ή NMR (400 MHz, DMSO-d6) δ 8.04 (s, 1H), 7.35 (s, 1H), 5.67 (s, 2H), 4.22 (t, J= 10.6 Hz, 2H), 4.14 (dd, J = 11.7, 2.6 Hz, 1H), 3.06 (t, J = 8.5 Hz, 1H), 2.39 - 2.26 (m, 2H), 2.23 2.16 (m, 3H), 2.12 (d, J = 13.6 Hz, 1H), 2.041.95 (m, 3H), 1.70 (dd, J = 13.6, 11.7 Hz, 1H), 1.32 (dd, J= 13.5, 11.2 Hz, 1H), 1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 470.15 (M+l)⁺
Compound 338	223; .	Ή NMR (400 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.35 (s, 1H), 4.53

340

Product	Starting Material and Amine	NMR, LCMS
1 ü V^^w c U- / \/~~\ X ^ZI n- _OA—/ z?~~2 ___/X.- Z' g	nh₂111··^ ho'	(t, J = 5.3 Hz, 1H), 4.35 (td, J = 7.7, 7.2, 2.2 Hz, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.14 (d, J = 11.1 Hz, 1H), 3.25 (td, J = 5.8, 1.6 Hz, 2H), 3.08 (s, 1H), 2.34 (d, J = 13.4 Hz, 1H), 2.13 (d, J = 13.6 Hz, 1H), 1.91 (dt, J = 13.5, 6.9 Hz, 1H), 1.70 (t, J= 12.6 Hz, 1H), 1.62 - 1.49 (m, 1H), 1.45 (dt, J = 13.7, 6.6 Hz, 1H), 1.39 - 1.23 (m, 1H), 1.04 (d, J = 6.2 Hz, 3H), 0.86 (d, J = 6.6 Hz, 3H). LCMS m/z 447.21 (M+l)⁺
Compound 339 E F °O>^ciΝ?νψ' N—^ N_x N	223; nh₂U N. N	*H NMR (400 MHz, DMSO-d6) δ 7.89 (s, 1H), 7.34 (s, 1H), 5.47 (s, 2H), 4.21 (t, J = 10.6 Hz, 2H), 4.11 (d, J = 11.4 Hz, 1H), 3.05 (s, lH),2.32(d, J = 13.1 Hz, 2H), 2.26 (s, 3H), 2.10 (d, J= 13.4 Hz, 1H), 1.76- 1.60 (m, 1H), 1.40- 1.24 (m, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 456.15 (M+l)⁺
Compound 340 E F °CQ“^ci Ν-Υΐ' N-J (/ /°	223; nh₂ o' /°	‘H NMR (400 MHz, DMSO-d6) δ 7.97 (s, 1H), 7.35 (s, 1H), 4.48 (t, J = 5.3 Hz, 2H), 4.23 (t, J = 10.6 Hz, 2H), 4.14 (dd, J= 11.7, 2.5 Hz, 1H), 3.79 (t, J = 5.2 Hz, 2H), 3.52 (dd, J = 5.9, 3.5 Hz, 2H), 3.41 (dd, J = 5.7, 3.6 Hz, 2H), 3.21 (s, 3H), 3.14-2.97 (m, 1H), 2.54 (s, 1H), 2.33 (d, J = 13.6 Hz, 1H), 2.13 (d, J= 13.7 Hz, 1H), 1.70 (dd, J= 13.6, 11.6 Hz, 1H), 1.33 (dd, J

341

Product	Starting Material and Amine	NMR, LCMS
		= 13.6, 11.3 Hz, 1H), 1.04 (d, J = 6.3 Hz, 3H). LCMS m/z 463.15 (M+l)⁺
Compound 341 E F °A>^CI N^J A	223; nh₂ /Ύ χΝ	Ή NMR (400 MHz, DMSO-d6) δ 8.03 (s, 1H), 7.35 (s, 1H), 7.06 (d, J=1.4 Hz,1H), 6.73 (d,.J = 1.3 Hz, 1H), 4.31 (t, J = 7.1 Hz, 2H), 4.23 (t, J = 10.7 Hz, 2H), 4.14 (d, J= 11.6 Hz, 1H), 3.86 (t, J = 7.1 Hz, 2H), 3.07 (s, 1H), 2.33 (d, J = 13.6 Hz, 1H), 2.22 (s, 5H), 2.14 (d, J = 13.6 Hz, 1H), 1.80- 1.62 (m, 1H), 1.35- 1.16 (m, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 483.24 (M+l)⁺
Compound 342 E F OA^ci AY* N-⁴ OH /°	223; nh₂p /°	Ή NMR (400 MHz, DMSO-d6) δ 7.90 (s, 1H), 7.35 (s, 1H), 5.28 (d, J = 5.5 Hz, 1H), 4.40 (dd, J =13.7, 3.3 Hz, 1H), 4.29-4.18 (m, 3H), 4.14 (dd, J= 11.5, 2.6 Hz, 1H), 3.95 (s, 1H), 3.29 - 3.22 (m, 5H), 3.07 (s, 1H), 2.33 (d, J= 13.6 Hz, 1H), 2.13 (d, J = 13.6 Hz, 1H), 1.71 (t, J = 12.6 Hz, 1H), 1.33 (dd, J = 13.5, 11.3 Hz, 1H), 1.04 (d, J = 6.2 Hz, 3H). LCMS m/z 449.15 (M+l)⁺
Compound 343	223; nh₂ O^	’H NMR (400 MHz, DMSO-d6) δ 7.89 (s, 1H), 7.35 (s, 1H), 4.46 4.28 (m, 2H), 4.28-4.18 (m, 2H), 4.12 (dd, J = 11.8, 2.6 Hz, 1H), 3.84 (d, J= 11.5 Hz, 1H), 3.65 (s, 1H), 3.28 (d, J = 10.7 Hz, 1H), 3.05 (d, J = 9.6 Hz, 1H), 2.34 (d, J

342

Product	Starting Material and Amine	NMR, LCMS
li '^z ^=/ XZ X X \ /\ / Tl *' \Z^ 2		= 13.4 Hz, 2H), 2.12 (d, J = 13.5 Hz, 1H), 1.72 (dd, J = 26.7, 14.3 Hz, 2H), 1.58 (d, J = 13.3 Hz, 1H), 1.51 - 1.27 (m, 4H), 1.19 (dd, J = 23.4, 11.5 Hz, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 459.18 (M+l)⁺
Compound 344 F. F oOQ“^ci M « N oM	223; nh₂ 0M	Ή NMR (400 MHz, DMSO-d6) δ 8.04 (s, 1H), 7.35 (s, 1H), 4.46 (t, J = 7.0 Hz, 2H), 4.23 (t, J= 10.7 Hz, 2H), 4.14 (d, J = 11.2 Hz, 1H), 3.18-3.03 (m, 3H), 3.00 (s, 3H), 2.34 (d, J = 13.6 Hz, 2H), 2.23 (p, J = 7.1 Hz, 2H),2.14(d, J = 13.6 Hz, 1H), 1.70 (dd, J = 13.5, 11.6 Hz, 1H), 1.32 (dd, J =13.6, 11.3 Hz, 1H), 1.04 (d, J = 6.3 Hz, 3H). LCMS m/z 481.08 (M+l)⁺
Compound 345 F F oÔQ*-⁰¹ M « N-J HN / H	223; nh₂oH	’H NMR (400 MHz, DMSO-d6) δ 7.92 (s, 1H), 7.88 (t, J = 5.6 Hz, 1H), 7.36 (s, 1H), 4.37 (t, J = 6.1 Hz, 2H), 4.29 -4.18(m, 2H), 4.13 (d, J = 11.5 Hz, 1H), 3.45 (q, J = 6.1 Hz, 2H), 3.07 (s, 1H), 2.35 - 2.23 (m, 3H), 2.14 (d, J= 13.6 Hz, 1H), 1.69 (dd, J= 13.6, 11.6 Hz, 1H), 1.38-1.25 (m, 1H), 1.03 (d, J = 6.2 Hz, 3H), 0.95 (dd, J = 6.9, 1.4 Hz, 6H). LCMS m/z 474.17 (M+l)⁺
Compound 346	223;	Ή NMR (400 MHz, DMSO-d6) δ 8.19 (dd, J = 4.8,1.6 Hz, 1H), 8.04 (s, 1H), 7.83 (dd, J = 7.9, 1.6 Hz, 1H), 7.34 (s, 1H), 7.05 (dd, J = 7.9,

343

Product	Starting Material and Amine	NMR, LCMS
F. F oÔQ-^cl ,N >/' Μ ^‘‘·ΐ, ^N. J H N-J $=N hn_vZA T N L Ù	nh₂T^Nhn_vXA I I	4.7 Hz, 1H), 5.75 (s, 2H), 4.21 (t, J = 10.6 Hz, 2H), 4.14 (d, J = 11.3 Hz, 1H), 3.05 (s, 1H), 2.40-2.28 (m, 1H), 2.10 (d, J = 13.5 Hz, 1H), 1.74- 1.60 (m, 1H), 1.32 (dd, J = 13.4, 11.4 Hz, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 492.14 (M+l)⁺
Compound 347 E ^F N-J 0 )—x ¹¹ // \\ —s—(' v n \—/ 0 —'	223; nh₂ o ;—x —s—(' v n \—/ 0 —'	Ή NMR (400 MHz, DMSO-d6) δ 8.08 (s, 1H), 8.01 (dd, J = 7.9, 1.5 Hz, 1H), 7.72 (td, J = 7.5, 1.5 Hz, 1H), 7.697.54 (m, 1H), 7.35 (s, 1H), 7.11 (d, J = 7.6 Hz, 1H), 6.04 (s, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.18 -4.03 (m, 1H), 3.31 (s, 3H), 3.06 (s, 1H), 2.43 2.30 (m, 2H), 2.12 (d, J = 13.5 Hz, 1H), 1.70 (dd, J= 13.6, 11.6 Hz, 1H), 1.46- 1.25 (m, 1H), 1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 529.12 (M+l)⁺
Compound 348 E F OA' N-J ( /=\ / f—\\ // ⁰ HO '—'	223; nh₂ ( /=\ / HO ^X—'	*H NMR (400 MHz, DMSO-d6) δ 7.90 (d, J = 7.1 Hz, 1H), 7.35 (s, 1H), 7.28 (dd, J = 8.8, 3.2 Hz, 2H), 6.95-6.84 (m, 2H), 5.71 (s, 1H), 4.90 (dd, J = 7.6, 4.9 Hz, 1H), 4.40 (dd, J = 6.2, 3.4 Hz, 2H), 4.23 (t, J = 10.6 Hz, 2H), 4.13 (d, J = 11.4 Hz, 1 H), 3.73 (d, J = 1.8 Hz, 3H), 3.06 (s, 1H), 2.33 (d, J = 30.9 Hz, 2H), 2.13 (d, J = 13.5 Hz, 1H), 1.73 - 1.60 (m, 1H), 1.32 (t, J= 12.5 Hz, 1H), 1.04 (d, J = 6.2 Hz, 3H). LCMS m/z 511.19 (M+l)⁺

344

Product	Starting Material and Amine	NMR, LCMS
. Compound 349 F. F oOlV⁰¹ O K-yÇ' N-J oJ N H	223; nh₂ oJ N H	Ή NMR (400 MHz, DMSO-d6) δ 7.97 (s, 1H), 7.70 (d, J = 8.1 Hz, 1H), 7.53 (d, J = 8.4 Hz, 1H), 7.34 (d, J = 7.2 Hz, 2H), 7.10 (t, J = 7.5 Hz, 1H), 5.91 (s, 2H), 4.20 (t, J =10.6 Hz, 2H), 4.09 (d, J = 11.6 Hz, 1H), 3.00 (s, 1H), 2.30 (d, J = 11.4 Hz, 2H), 2.08 (d, J = 13.6 Hz, 1H), 1.741.62 (m, 1H), 1.38 - 1.16 (m, 1H), 0.99 (d, J = 6.2 Hz, 3H). LCMS m/z 491.15 (M+l)⁺
Compound 350 F F oO3~^ci Mr Ν'⁴ n'xAnh \=N	223; nh₂ n'XC-NH \=N	’H NMR (400 MHz, DMSO-d6) δ 12.29 (s, 1H), 8.70 (s, 1H), 8.09 (s, 1H), 7.56 (d, J = 3.6 Hz, 1H), 7.35 (s, 1H), 6.28 (d, J = 3.6 Hz, 1H), 5.96 (s, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.16 (d, J = 11.3 Hz, 1H), 3.06 (s, 1H), 2.35 (d, J= 13.2 Hz, 2H),2.12(d, J = 13.6 Hz, 1H), 1.82-1.64 (m, 1H), 1.33 (dd, J = 13.6, 11.2 Hz, 1H), 1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 492.14 (M+l)⁺
Compound 351 F. F O>^C\| J	223; nh₂	‘H NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.35 (s, 1H), 6.93 (s, 1H), 6.85 (s, 1H), 4.66 (t, J = 7.4 Hz, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.11 (d, J = 11.4 Hz, lH),3.19(t, J = 7.4 Hz, 2H), 3.05 (s, 1H), 2.40 (s, 3H), 2.29 (d, J = 11.4 Hz, 2H), 2.21 (s, 3H), 2.13 (d, J = 13.6 Hz, 1H), 1.67 (dd, J= 13.6, 11.6 Hz, 1H), 1.31 (dd, J = 13.5, 11.3 Hz, 1H),

345

Product	Starting Matériel and Amine	NMR, LCMS
		1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 494.21 (M+l)⁺
Compound 352 E F oôi3-^cl N χ,χ''' κι A/,, «J H N-⁴	223; nh₂	Ή NMR (400 MHz, DMS0-d6) δ 8.74 (d, J = 2.1 Hz, 1H), 8.13 (s, 1H), 8.07 (d, J = 8.0 Hz, 1H), 7.92 (dd, J = 8.0, 2.2 Hz, 1H), 7.34 (d, J = 5.9 Hz, 1H), 5.75 (s, 2H), 4.22 (t, J =10.6 Hz, 2H), 4.14 (d, J= 11.2 Hz, 1H), 3.06 (s, 1H), 2.42-2.30 (m, 2H), 2.12 (d, J = 13.3 Hz, 1H), 1.68 (dd, J =13.5, 11.6 Hz, 1H), 1.38 - 1.28 (m, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 477.15 (M+l)⁺
Compound 353 E F OQ~^ci N⁴ -¥	223; nh₂A	Ή NMR (400 MHz, DMSO-d6) δ 7.96 (s, 1H), 7.35 (s, 1H), 5.40 (s, 2H), 4.21 (t, J= 10.6 Hz, 2H), 4.11 (d, J = 11.5 Hz, 1H), 3.05 (s, 1H), 2.44 (s, 3H), 2.40 2.28 (m, 2H), 2.14 (s, 4H), 1.67 (dd, J= 13.6, 11.6 Hz, 1H), 1.30 (dd, J = 13.6, 11.3 Hz, 1H), 1.02 (d, J = 6.3 Hz, 3H). LCMS m/z 470.15 (M+l)⁺
Compound 354 F. F <O>^C\|K-yV' N-J (/ ^F	223; nh₂ 0^ ^F	’H NMR (400 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.35 (s, 1H), 7.16 (dddd, J= 19.9, 17.9, 11.2, 4.8 Hz, 3H), 7.026.90 (m, 1H), 4.75 (t, J = 5.1 Hz, 2H), 4.47 (t, J = 5.1 Hz, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.14 (d, J = 11.6 Hz, 1H), 3.06 (s, 1H), 2.32 (d, J= 10.6 Hz, 2H), 2.13 (d, J = 13.5 Hz, 1H), 1.69 (dd, J = 13.7, 11.6 Hz, 1H),

346

Product	Starting Material and Amine	NMR, LCMS
		1.33 (dd, J = 13.6, 11.3 Hz, 1H), 1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 499.18 (M+l)⁺
Compound 355 F F oQQ~^ci <^ν3Υ' N-J Λ°	223; nh₂ Λ”	‘H NMR (400 MHz, DMSO-d6) δ 8.10 (s, 1H), 7.99 - 7.90 (m, 2H), 7.54 (d, J = 8.3 Hz, 2H), 7.35 (s, 1H), 5.70 (s, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.14 (d, J = 11.7 Hz, 1H), 3.21 (s, 3H), 3.06 (s, 1H), 2.35 (d, J = 15.3 Hz, 2H), 2.12 (d, J = 13.5 Hz, 1H), 1.741.63 (m, 1H), 1.31 (dd, J = 13.5, 11.3 Hz, 1H), 1.02 (d, J = 6.2 Hz, 3H). LCMS m/z 529.16 (M+l)⁺
Compound 356 F F o0c*-^ci N⁴ O^O	223; nh₂Jo	Ή NMR (400 MHz, DMSO-d6) δ 7.86 (s, 1H), 7.35 (s, 1H), 5.38 (s, 2H), 4.23 (t, J = 10.6 Hz, 2H), 4.15 (d, J = 11.6 Hz, 1H), 3.45 (d, J = 8.8 Hz, 4H), 3.07 (s, 1H), 2.34 (d, J = 13.4 Hz, 2H), 2.13 (d, J = 13.6 Hz, 1H), 1.71 (dd, J = 13.5, 11.6 Hz, 1H), 1.59 (q, J = 6.5, 5.6 Hz, 4H), 1.45 (s, 2H), 1.41 1.26 (m, 1H), 1.04 (d, J = 6.3 Hz, 3H). LCMS m/z 429.15 (M+l)⁺
Compound 357 F. F oOtA¹ N?yY' N-⁴	223; nh₂	‘H NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.35 (s, 1H), 4.75 (s, 1H), 4.29 (s, 2H), 4.23 (t, J = 10.6 Hz, 2H), 4.14 (d, J = 11.5 Hz, 1H), 3.15-3.02 (m, 3H), 2.35 (d, J =12.3 Hz, 2H), 2.12 (d, J =13.5 Hz, 1H), 1.70 (dd, J = 13.7, 11.7 Hz, 1H), 1.34

347

Product	Starting Material and Amine	NMR, LCMS
		(dd, J =13.6, 11.3 Hz, 1H), 1.04 (d, J = 6.3 Hz, 3H), 0.62 (q, J = 4.0 Hz, 2H), 0.48 (q, J = 4.1 Hz, 2H). LCMS m/z 445.18 (M+l)⁺
Compound 358 F. ^Fοόί>“ <y^-T^% Ν'' A	223; nh₂ N	’H NMR (400 MHz, DMSO-d6) δ 9.12 (d, J = 1.5 Hz, 1H), 8.61 (dd, J = 4.7, 1.6 Hz, 1H), 8.07 (s, 1H), 8.02 (d, J = 4.7 Hz, 1H), 7.97 (s, 1H), 7.34 (s, 1H), 6.09 (s, 2H), 4.20 (t, J= 10.6 Hz, 2H), 4.09 (d, J = 11.3 Hz, 1H), 3.01 (d, J = 11.7 Hz, 1H), 2.30 (d, J = 15.4 Hz, 2H), 2.10 (d, J = 13.6 Hz, 1H), 1.64 (dd, J= 13.5, 11.7 Hz, 1H), 1.38- 1.19 (m, 1H), 1.00 (d, J = 6.2 Hz, 3 H). LCMS m/z 491.18 (M+l)⁺
Compound 359 F. F <OQ-^ci N J OH	223; nh₂OH	Ή NMR (400 MHz, DMSO-d6) δ 7.97 (s, 1H), 7.35 (s, 1H), 4.67 (s, 1H), 4.36 (t, J = 7.1 Hz, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.13 (d, J = 11.4 Hz, 1H), 3.39 (s, 2H), 3.07 (s, 1H), 2.34 (s, 2H), 2.13 (d, J= 13.8 Hz, 1H), 1.93 (p, J = 6.5 Hz, 2H), 1.70 (dd, J = 13.6, 11.6 Hz, 1H), 1.32 (dd, J= 13.6, 11.3 Hz, 1H), 1.03 (d, J = 6.3 Hz, 3H). LCMS m/z 419.13 (M+l)⁺
Compound 360	223;	‘H NMR (400 MHz, DMSO-d6) δ 8.23 (d, J = 2.5 Hz, 1H), 8.03 (s, 1H), 7.69 (dd, J = 8.6, 2.5 Hz, 1H), 7.34 (s, 1H), 6.83 (d, J = 8.5 Hz, 1H), 5.51 (s, 2H), 4.21 (t, J = 10.6 Hz, 2H), 4.11

34S

Product	Starting Material and Amine	NMR, LCMS
ο V^Xtfî >' u. / \/ \ >( X ^ZIq\-/ Z z^zx/°x Z '—	nh₂S z°	(d, J = 11.4 Hz, 1H), 3.83 (s, 3H), 3.04 (s, 1H), 2.32 (d, J =13.7 Hz, 2H), 2.11 (d, J = 13.6 Hz, 1H), 1.75-1.58 (m, 1H), 1.30 (dd, J = 13.5, 11.2 Hz, 1H), 1.01 (d, J = 6.2 Hz, 3H). LCMS m/z 482.16 (M+l)⁺
Compound 361 F. F oCQ-^ci ΛΤ' N-J HC)	223; nh₂ HC)	’H NMR (400 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.35 (s, 1H), 5.03 (s, 1H), 4.35 (t, J = 5.4 Hz, 2H), 4.22 (t, J= 10.6 Hz, 2H), 4.14 (d, J = 11.6 Hz, 1H), 3.76 (t, J = 5.1 Hz, 2H), 3.07 (s, 1H), 2.33 (d, J= 14.0 Hz, 2H), 2.13 (d, J = 13.6 Hz, 1H), 1.70 (dd, J = 13.6, 11.6 Hz, 1H), 1.32 (dd, J = 13.6, 11.2 Hz, 1H), 1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 405.13 (M+l)⁺
Compound 362 F F oOq*-^ciA V' ^J^~F /°	223; nh₂ F 0 /	Ή NMR (400 MHz, DMSO-d6) δ 8.01 (s, 1H), 7.34 (s, 1H), 7.28 7.21 (m, 1H), 7.20-7.10 (m, 2H), 5.47 (s, 2H), 4.21 (t, J= 10.6 Hz, 2H), 4.11 (d, J = 11.8 Hz, 1H), 3.82 (s, 3H), 3.02 (d, J = 19.8 Hz, 1H), 2.33 (d, J= 13.6 Hz, 2H), 2.11 (d, J = 13.5 Hz, 1H), 1.66 (dd, J = 13.6, 11.6 Hz, 1H), 1.31 (dd, J = 13.6, 11.3 Hz, 1H), 1.01 (d, J = 6.3 Hz, 3H). LCMS m/z 499.18 (M+l)⁺
Compound 363	223;	^!H NMR (400 MHz, DMSO-d6) δ 7.90 (s, 1H), 7.35 (s, 1H), 5.14

349

Product	Starting Material and Amine	NMR, LCMS
---ο ΊΓ ^z ''z Z i l , o Z \ r\fv IZ X X \ /\ / -n o	NH, c OH	(s, 1H), 4.86 (s, 1H), 4.45 (dd, J =13.8, 3.6 Hz, 1H), 4.22 (dd, J = 13.3, 8.9 Hz, 3H), 4.14 (d, J = 14.4 Hz, 1H), 3.79 (s, 1H), 3.26 (s, 2H), 3.07 (s, 1H), 2.33 (d, J = 14.2 Hz, 2H), 2.13 (d, J= 13.6 Hz, 1H), 1.70 (dd, J = 13.5, 11.6 Hz, 1H), 1.38-1.27 (m, 1H), 1.04 (d, J = 6.3 Hz, 3H). LCMS m/z 435.16 (M+l)⁺
Compound 364 F. F °OQ~^ci _nJA N-J OH	223; nh₂	‘H NMR (400 MHz, DMSO-d6) δ 7.85 (s, 1H), 7.35 (s, 1H), 4.83 (s, 1H), 4.24 (d, J = 12.4 Hz, 4H), 4.14 (d, J = 11.5 Hz, 1H), 3.07 (s, 1H), 2.35 (d, J = 13.8 Hz, 2H),2.12(d, J = 13.6 Hz, 1H), 1.70 (dd, J = 13.6, 11.6 Hz, 1H), 1.34 (dd, J = 13.6,11.2 Hz, 1H), 1.08-1.01 (m, 9H). LCMS m/z 433.17 (M+l)⁺
Compound 365 F. F oQQ~^ci JA' ^N. J H N-J h N	223; nh₂h N	Ή NMR (400 MHz, DMSO-d6) δ 9.09 (s, 1H), 8.05 (s, 1H), 7.99 (s, 1H), 7.34 (s, 1H), 5.88 (s, 2H), 4.21 (t, J = 10.6 Hz, 2H), 4.12 (d, J = 11.0 Hz, 1 H), 3.05 (s, 1H), 2.33 (d, J= 13.6 Hz, 2H), 2.11 (d, J = 13.7 Hz, 1H), 1.67 (dd, J = 13.5, 11.6 Hz, 1H), 1.31 (dd, J= 13.5, 11.3 Hz, 1H), 1.02 (d, J = 6.3 Hz, 3H). LCMS m/z 458.10 (M+l)⁺
Compound 366	223;	*H NMR (400 MHz, DMSO-d6) δ 10.68 (s, 2H), 7.96 (s, 1H), 7.34 (s, 1H), 6.95 (dd, J = 7.9, 1.6 Hz, 1H), 6.92 (d,

350

Product	Starting Material and Amine	NMR, LCMS
ο γσ, ¢- \—/ X u.^ / \/ \ o XJQ¹ H -YjU Z ^x—	nh₂ N^OH	J =1.5 Hz, 1H), 6.89 (d, J = 7.9 Hz, 1H), 5.49 (s, 2H), 4.21 (t, J = 10.6 Hz, 2H), 4.10 (d, J = 11.5 Hz, 1H), 3.02 (d, J = 16.3 Hz, 1H), 2.33 (t, J = 13.7 Hz, 2H),2.10(d, J = 13.4 Hz, 1H), 1.66 (dd, J= 13.6, 11.6 Hz, 1H), 1.35 - 1.23 (m, 1H), 1.01 (d, J = 6.2 Hz, 3H). LCMS m/z 507.17 (M+l)⁺
Compound 367 F. F O>^CI N-J 4 OH	223; nh₂ 4 OH	Ή NMR (400 MHz, DMSO-d6) δ 7.99 (s, 1H), 7.35 (s, 1H), 4.52 (s, 1H), 4.38 (dd, J = 7.0, 4.2 Hz, 2H), 4.22 (t, J = 10.6 Hz, 2H), 4.12 (d, J= 11.2 Hz, 1H), 3.06 (s, 1H), 2.32 (d, J = 11.6 Hz, 2H), 2.13 (d, J = 13.5 Hz, 1H), 1.951.83 (m, 2H), 1.69 (dd, J = 13.6, 11.7 Hz, 1H), 1.32 (dd, J =13.6, 11.2 Hz, 1 H), 1.13 (s, 6H), 1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 447.17 (M+l)⁺
Compound 368 F. F oOQ-⁰¹ ^NJ H N-J Ç NH o	223; nh₂ ( NH O	‘H NMR (400 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.44 (s, 1H), 7.35 (s, 1H), 4.33 (d, J = 7.2 Hz, 2H), 4.23 (t, J= 10.6 Hz, 2H), 4.13 (d, J = 11.4 Hz, 1H), 3.12-2.96 (m, 2H), 2.89 (t, J = 10.8 Hz, 1H), 2.34 (d, J = 13.0 Hz, 2H), 2.30-2.05 (m,4H), 1.69 (t, J = 12.5 Hz, 2H), 1.43 (t, J = 9.9 Hz, 1H), 1.39- 1.28 (m, 1H), 1.03 (d, J = 6.2 Hz, 3H). LCMS m/z 472.18 (M+l)⁺
Compound 369	223;	Ή NMR (400 MHz, DMSO-d6) δ 8.03 (d, J

351

Product	Starting Material and Amine	NMR, LCMS
o IL / \/ \ X zx —/ •Z.	nh₂	= 90.8 Hz, 1H), 7.35 (s, 1H), 5.40 (d, J = 306.9 Hz, 2H), 4.19 (dt, J = 29.2, 9.8 Hz, 3H), 3.07 (s, 1H), 2.43 (s, 1H), 2.35 (d, J = 13.6 Hz, 1H), 2.13 (d, J= 13.5 Hz, 1H), 1.71 (t, J = 12.6 Hz, 1H), 1.40- 1.29 (m, 1H), 1.04 (d, J = 6.2 Hz, 3H). LCMS m/z 400.12 (M+l)⁺
Compound 370 F. F oCjQ-^ci <7 h N-⁴ DH	223; nh₂ DH	Ή NMR (400 MHz, DMSO-d6) δ 7.94 (s, 1H), 7.35 (s, 1H), 4.73 (t, J = 5.2 Hz, 1H), 4.35 (dd, J = 13.5, 5.9 Hz, 1H), 4.22 (t, J = 10.6 Hz, 2H), 4.17-4.07 (m, 2H), 3.30-3.19 (m, 2H), 3.08 (d, J = 9.7 Hz, 1H), 2.34 (d, J = 13.5 Hz, 2H), 2.17-1.99 (m, 2H), 1.70 (dd, J= 13.5, 11.6 Hz, 1H), 1.33 (dd, J = 13.6, 11.1 Hz, 1H), 1.03 (d, J = 6.2 Hz, 3H), 0.77 (d, J = 6.8 Hz, 3H). LCMS m/z 433.17 (M+l)⁺
Compound 371 F. F oOt>^cl N-J »1111 OH	223; nh₂OH	Ή NMR (400 MHz, DMSO-d6) δ 7.90 (s, 1H), 7.35 (s, 1H), 5.12 (s, 1H), 4.84 (s, 1H), 4.45 (dd, J =13.9, 3.5 Hz, 1H), 4.27-4.10 (m, 4H), 3.79 (s, 1H), 3.07 (t, J = 7.9 Hz, 1H), 2.34 (d, J= 13.4 Hz, 1H), 2.13 (d, J= 13.6 Hz, 1H), 1.71 (dd, J= 13.5, 11.7 Hz, 1H), 1.39-1.28 (m, 1H), 1.04 (d, J = 6.2 Hz, 3H). LCMS m/z 435.16 (M+l)⁺

^xSharpless, K. B. et al. Nature, 2019, 574, 86-89

352

Préparation of S56 methyl 1-[(2S, 6S)-6-methyl-4-oxo-2-piperidyl]cyclopropanecarboxylate (S56)

L-Proline, Et₃N, MgSO₄EtOH

Standard Method C: Cyclization to Préparé Piperidone with Distillation Workup

A stirred solution of (45)-4-aminopentan-2-one (hydrochloric acid) S25 (1.25 g, 8.2 mmol) in EtOH (50 mL) was cooled to 0 °C. Then Et₃N (2.28 g, 3.2 mL, 22.0 mmol), MgSO₄(950 mg, 7.7 mmol), L-Proline (490 mg, 4.0 mmol) and methyl 1formylcyclopropanecarboxylate C124 (1 g, 7.6 mmol) were added. After 30 minutes, the reaction mixture was slowly allowed to warm to room température and stirred for 24 hours. The reaction mixture was distilled under vacuum to give crude product as a brown color thick oil. The crude material was purified by silica gel flash column chromatography (eluted with 12 % MeOH in DCM) to afford the product methyl l-[(2S,6S)-6-methyl-4-oxo-2piperidyl]cyclopropanecarboxylate S56 (600 mg, 37% yield) as a pale yellow oil. Partial stereochemical érosion of the enantiomerically pure starting material (4S)-4-aminopentan-2-one (Hydrochloride sait) S25 was observed under these conditions, leading to unseparated mixtures of stereoisomers where the cis-product was the major isomer. 'H NMR (400 MHz, DMSO-d6) δ 3.58 (s, 3H), 2.90-2.86 (m, 1H), 2.81-2.72 (m, 1H), 2.30-2.13 (m, 3H), 2.06-1.97 (m, 1H), 1.100.94 (m, 8H). LCMS m/z 212.1 [M+H]⁺.

Préparation ofS57 tert-butyl N-[1 ~[(2S, 6S)-6-methyl-4-oxo-2-piperidyl]cyclopropyl]carbamate (S57)

C125

S25

L-Proline, Et₃N, MgSO₄EtOH

S57

Standard Method D: Cyclization to Préparé Piperidone with Aquêous Quench Workup

To a stirred solution of (4S)-4-aminopentan-2-one (hydrochloric acid) (1 g, 6.5 mmol)

353

S25 in EtOH (20 mL) were added L-proline (150 mg, 1.3 mmol), MgSCU (783 mg, 6.4 mmol), EtsN (718.74 mg, 1 mL, 7.0 mmol) and tert-butyl N-(l-formylcyclopropyl)carbamate C125 (1.2 g, 6.4 mmol) at room température. The reaction mass was allowed to stir for 16 hours at room température. The reaction mixture was diluted with saturated NaHCCh (50 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layer was washed with 0.15 N aqueous HCl (3 x 50 mL), and then the aqueous layer pH was adjusted to 12 by using 1 N NaOH solution. The aqueous layer extracted with EtOAc (3x100 mL), and the combined organic layer was dried over Na₂SO4, fïltered, and evaporated under vacuum to provide the crude material. The crude material was purified by column chromatography using 100-200 mesh silica gel and eluted with 4% MeOH in DCM to provide tert-butyl N-[l-[(2S,6S)-6-methyl-4-oxo-2piperidyl] cyclopropyl] carbamate S57 (800 mg, 46% yield). Partial stéréo Chemical érosion of the enantiomerically pure starting material (4S)-4-aminopentan-2-one (Hydrochloride sait) S25 was observed under these conditions, leading to unseparated mixtures of stereoisomers where the cisproduct was the major isomer. *H NMR (400 MHz, DMSO-d6) δ 7.11 (brs, 1H), 2.74 (brs, 1H), 2.49 (brs, 1H), 2.18-2.0 (m, 5H), 1.36 (s, 9H), 1.09 (d, J=6.0 Hz, 3H), 0.75-0.60 (m, 4H). LCMS m/z 269.2 [M+H]⁺.

Préparation of S58 tert-butyl N-[l-[(2S, 6S)-6-methyl-4-oxo-2-piperidyl]cyclopropyl]carbamate (S58)

^C126 C127 C128 S58

Step 1. l-(dimethylamino)-N-methoxy-N-methyl-cyclopropanecarboxamide (C128)

To a stirred solution of l-(dimethylamino)cyclopropanecarboxylic acid C126 (500 mg, 0.0038 mol) in DCM (10 mL) were added DIPEA (2.226 g, 3.00 mL, 0.0169 mol) and N-methoxymethanamine (hydrochloric acid) C127 (453 mg, 0.0046 mol) at 0 °C. The reaction mixture was stirred for 10 minutes, and T3P (3.12 g, 2.92 mL of 50 %w/w, 0.0049 mol) was slowly added at 0 °C. The reaction mixture was stirred at room température for 16 hours. The reaction mixture was diluted with DCM (25 mL) and cooled to 0 °C. 1 N NaOH solution (10 mL) was slowly added, and the organic layer was separated and washed with saturated NH4CI solution (10 mL). The organic layer was dried over anhydrous sodium sulfate, fïltered, and concentrated under reduced pressure to obtained the crude material. The crude material was purified by column chromatography using 100-200 mesh silica gel and eluted with 70% EtOAc 354 in petroleum ether to afford l-(dimethylamino)-N-methoxy-N-methyl-cyclopropanecarboxamide C128 (300 mg, 44%) as a brown color liquid. ’H NMR (400 MHz, DMSO-d6) δ 3.62 (s, 3H), 3.19 (s, 3H), 2.23 (s, 6H), 0.82-0.80 (m, 4H). LCMS m/z 173.22 [M+H]⁺.

Step 2. l-(dimethylamino)cyclopropanecarbaldehyde (S58)

To a stirred suspension of LAH (988 mg, 0.0255 mol) in diethyl ether (50 mL) was added l-(dimethylamino)-N-methoxy-N-methyl-cyclopropanecarboxamide C128 (2 g, 0.0102 mol) dropwise at 0 °C. The reaction mixture was allowed to stir for 4 hours at 0 °C. The reaction was quenched with water (3.2 mL), 1 N NaOH solution (3.2 mL), and water (3.2 mL). The reaction mass was filtered through a Celite® pad and washed with diethyl ether (30 mL). The filtrate was evaporated under vacuum to afford crude l-(dimethylamino)cyclopropanecarbaldehyde S58 (1.4 g, 97%) as a colorless oil. *H NMR (300 MHz, Chloroform-d) δ 9.01 (s, 1H), 2.63 (s, 6H), 1.281.14 (m, 4H).

Intermediates S59-S62

Intermediates S59-S62 (see Table 12) were prepared in a single step from intermediate S25 using standard Method C or D. Corresponding aldéhydes were prepared by methods described above or obtained from commercial sources. Partial stereochemical érosion of the enantiomerically pure starting material (4S)-4-aminopentan-2-one (hydrochloride sait) S25 was observed in step 1 leading to unseparated mixtures of stereoisomers being generated in Step 1. In each case the cis-product was the major isomer. Any modifications to methods are noted in Table 12 and accompanying footnotes.

Table 12. Structure and physicochemical data for intermediates S59-S62

Intermediate	Structure	Aldéhyde Reagent	Method	Ή NMR; LCMS m/z [M+HI
S59	1 ⁰ Aa	O y°\ O '—	C	‘H NMR (400 MHz, DMSO-d6) δ 4.09-4.01 (m, 2H), 2.78-2.65 (m, 1H), 2.39-2.31 (m, 1H), 2.25-1.97 (m,4H), 1.681.60 (m, 1H), 1.43-1.32 (m, 1H), 1.20-1.18 (m, 6H), 1.11-0.87 (m, 3H)

355

Intermediate	Structure	Aldéhyde Reagent	Method	^]HNMR; LCMS m/z. IM+Hf
S60	0 °\	\> <χ₀	D	Ή NMR (400 MHz, DMSO-d6) δ 4.09-4.01 (m, 2H), 2.78-2.65 (m, 1H), 2.39-2.31 (m, 1H), 2.25-1.97 (m,4H), 1.681.60 (m, 1H), 1.43-1.32 (m, 1H), 1.20-1.18 (m, 6H), 1.11-0.87 (m, 3H); 198.1
S61	OOP CN ^H		D	‘H NMR (400 MHz, DMSO-d6) δ 2.80-2.75 (m, 2H), 2.33-2.22 (m, 3H), 2.13-2.11 (m, 2H), 1.23-1.00 (m, 7H); 179.2
S62	A /ir	/ ⁰E	D¹	‘H NMR (400 MHz, DMSO-d6) δ 4.09-4.01 (m, 2H), 2.78-2.65 (m, 1H), 2.39-2.31 (m, 1H), 2.25-1.97 (m, 4H), 1.681.60 (m, 1H), 1.43-1.32 (m, 1H), 1.20-1.18 (m, 6H), 1.11-0.87 (m, 3H); 198.1

Note:

1) The workup procedure was modified as: the reaction mixture was diluted with saturated NaHCCh (50 mL) and extracted with EtOAc (4 x 300 mL). The combined organic layer was dried over sodium sulfate, filtered, and evaporated under vacuum to provide the crude material.

Compound 372 methyl l-[(2'S, 6'S, 7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]~ '-yl] cyclopropanecarboxylate (372)

356

Standard Method E: To Préparé Spiropiperidine

To a mixture ofmethyl l-[(2S,6S)-6-methyl-4-oxo-2-piperidyl]cyclopropanecarboxylate S56 (100 mg, 0.473 mmol) and 2-(5-chloro-3-thienyl)ethanol S2 (92 mg, 0.57 mmol, 1.2 equiv) in DCM (2.2 mL) was added MsOH (250 pL, 3.85 mmol, 8 equiv), and the mixture was heated to 40 °C. After 5 hours, the reaction was quenched with saturated NaHCCh solution and extracted with DCM (6x). The combined organic layer was dried over Na₂SO4, filtered, and concentrated. The crude material was purified with a silica gel column and eluted with 0 to 100% EtOAc in heptane to provide methyl l-[(2'S,6'S,7S)-2-chloro-6'-methyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-2'-yl]cyclopropanecarboxylate 372 (123 mg, 69%). Due to partial érosion of enantiomeric purity when preparing S56, 372 was also isolated as a mixture of enantiomers where the (2'S,6'S,7S) configuration was the major enantiomer. ’H NMR (300 MHz, Chloroform-7) δ 6.56 (s, 1H), 3.88 (t, J = 5.5 Hz, 2H), 3.63 (s, 3H), 3.04 (dtd, J = 12.9, 6.5, 2.5 Hz, 1H), 2.73 (dd, J = 11.8, 2.5 Hz, 1H), 2.58 (t, J = 5.5 Hz, 2H), 2.00 (ddt, J = 17.5, 13.4, 2.5 Hz, 2H), 1.70 (dd, J = 13.3, 11.7 Hz, 2H), 1.40 -1.16 (m, 3H), 1.11 (d, J = 6.4 Hz, 3H), 1.04 - 0.92 (m, 1H), 0.77 (ddd, J = 9.7, 5.4, 2.6 Hz, 1H). LCMS m/z 356.15 [M+H]⁺.

Compound 373-378

Compounds 373-378 (see Table 13) were prepared in a single step from the corresponding piperidinone reagent using standard method E. Due to partial érosion of enantiomeric purity when preparing piperidone reagents, compounds were isolated as a mixture of enantiomers where the (2'S,6'S,7S) configuration was the major enantiomer. Any modifications to methods are noted in Table 13 and accompanying footnotes.

357

Table 13. Structure and physicochemical data for compounds 373-378

Compound	Structures	Piperidone Reagent	¹II NMR ; LCMS m/z [M+H]⁺
373	x N) Z /—\ °M A / ⁷ AA ο	Y a Λ H	Ή NMR (300 MHz, Chloroform-tT) δ 6.58 (s, 1H), 3.88 (t, J = 5.5 Hz, 2H), 3.17 (dd, J =10.3, 5.1 Hz, 1H), 2.59 (t, J = 5.5 Hz, 2H), 2.34 (dd, J= 11.8, 2.3 Hz, 1H), 2.09- 1.93 (m, 2H), 1.62 (d, J= 12.7 Hz, 1H), 1.45 - 1.28 (m, 1H), 1.14 (d, J = 6.3 Hz, 3H), 0.71 - 0.60 (m, 1H), 0.60 0.45 (m, 2H), 0.45 - 0.37 (m, 1H); 356.15
374¹	\ » f/ o \ )—\ M IZ Y \ 1 Q	1 ⁰ AjA	‘H NMR (300 MHz, Chloroformé δ 8.40 (s, 1H), 6.59 (s, 1H), 4.10 (qd, J = 7.2, 2.7 Hz, 2H), 3.86 (t, J = 5.5 Hz, 2H), 3.29 (d, J = 12.3 Hz, 1H), 2.66 - 2.46 (m, 3H), 2.24- 1.82 (m, 3H), 1.81-1.68 (m, 2H), 1.68 - 1.54 (m, 1H), 1.301.06 (m, 7H), 0.87-0.69 (m, 1H); 370.31
375¹	'^ξ—o \ » o \ )—\ M iz Y \ ^ω\Μ O	1 ⁰ V H	Ή NMR (300 MHz, Chloroformé δ 8.43 (s, 1H), 8.38 (s, 1H), 6.60 (s, 1H), 4.08 (qd, J = 7.1, 2.3 Hz, 2H), 3.87 (td, J = 5.6, 3.1 Hz, 2H), 3.70 - 3.28 (m, 2H), 2.78 (d, J = 10.8 Hz, 1H), 2.61 (q, J = 5.0, 4.4 Hz, 2H), 2.38 -2.07 (m, 3H), 1.95 (t, J = 13.3 Hz, 1H), 1.70 (s, 1H), 1.53 (dt, J = 8.9, 4.4 Hz, 1H), 1.39 (dd, J = 6.5, 1.7 Hz, 3H), 1.33 0.97 (m, 5H); 370.31

358

Compound	Structures	Piperidone Reagent	’HNMR; LCMS m/z [M+H]⁺
376²	θ' IZ Y \ O	0 xj^A °\	’H NMR (300 MHz, Chloroformé) δ 8.44 (d, J = 1.9 Hz, 2H), 6.61 (s, 1H), 3.87 (t, J = 5.4 Hz, 2H), 3.78 - 3.45 (m, 2H), 3.38 (d, J = 3.0 Hz, 3H),3.15(d, J = 12.2 Hz, 2H), 2.61 (t, J = 5.4 Hz, 2H),2.16(td, J = 10.4, 9.0, 2.8 Hz, 2H), 2.05 (d, J = 13.4 Hz, 1H), 1.82 (t, J = 13.2 Hz, 1H), 1.38 (d, J = 6.6 Hz, 3H), 1.02 (s, 1H), 0.65 (p, J = 5.6 Hz, 2H), 0.51 (t, J = 9.5 Hz, 1H); 342.32
377²	oCcvyxs _CNf Ί V H	0 [>Xa CN	‘H NMR (300 MHz, Chloroformé) δ 8.26 (s, 2H), 6.60 (s, 1H), 4.13 3.60 (m, 2H), 3.44 (ddd, J = 12.0, 6.5, 2.8 Hz, 1H), 2.75 (dd, J = 12.0, 2.8 Hz, 1H), 2.61 (q, J = 5.2 Hz, 2H), 2.26 (dt, J = 14.2, 2.7 Hz, 1H), 2.19-2.01 (m, 2H), 1.76 (dd, J= 14.4, 11.9 Hz, 1H), 1.53 - 1.32 (m, 3H), 1.30 (d, J = 6.4 Hz, 3H), 1.06- 0.82 (m, 1H); 323.26
378³	\ IZ X / O	x’i	Ή NMR (400 MHz, Methanol-<4) δ 6.75 (s, 1H), 3.95 (t, J = 5.5 Hz, 2H), 3.70 (ddt, J =13.3, 6.8,3.4 Hz, 1H), 3.21 (d, J =12.2 Hz, 1H), 2.64 (td, J = 5.4, 1.5 Hz, 2H), 2.42 (s, 6H), 2.30 (ddt, J = 14.4, 9.2, 2.7 Hz, 2H), 1.98 (dd, J= 14.5, 12.4 Hz, 1H), 1.80 (dd, J = 14.9, 12.2 Hz, 1H), 1.39 (d, J = 6.6 Hz, 3H), 1.27-1.04 (m, 2H), 0.93 (dd, J= 11.3, 4.8 Hz, 1H), 0.75 (dd, J = 11.0, 5.0 Hz, 1H); 341.37

Note:

1) Compounds 374 and 375 were separated with silica gel chromatography and eluted with 0% to

359

70% EtOAc/EtOH (3:1) in heptane, followed by reversed-phase HPLC. Method: Waters XSelect CSH Cl8 OBD Prep Column; 30 x 150 mm, 5 micron. Gradient: acetonitrile in water with 0.2% formic acid.

2) Purification by reversed-phase HPLC. Method: Waters XSelect CSH Cl8 OBD Prep Column; 30 x 150 mm, 5 micron. Gradient: acetonitrile in water with 0.2% formic acid.

3) Purification by reversed-phase HPLC. Method: Waters XSelect CSH Cl8 OBD Prep Column;

x 150 mm, 5 micron. Gradient: acetonitrile in water with 0.1% trifluoroacetic acid.

Compound 379

N-[1 -[(2'S, 6'S, 7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-2'yl]cyclopropyl]acetamide (379)

At 0 °C, to a mixture of l-[(2'S,6'S,7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-2'-yl]cyclopropanamine 373 (14.5 mg, 0.044 mmol) in DCM (200 pL) was added N,N-diethylethanamine (20 pL, 0.13 mmol) and acetyl chloride (3 pL, 0.04 mmol). After 30 minutes, the reaction was quenched with saturated NaHCCh solution and extracted with EtOAc (4x). The combined organic layer was dried over Na2SO4, filtered, and concentrated. Purification by reversed-phase HPLC (Method: Waters XSelect CSH Cl8 OBD Prep Column; 30 x 150 mm, 5 micron. Gradient: acetonitrile in water with 5 mM hydrochloric acid) afforded the product N-[l-[(2'S,6'S,7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-2'-yl]cyclopropyl]acetamide (hydrochloride sait) 379 (4.9 mg, 27% yield). Due to partial érosion of enantiomeric purity when preparing 373, 379 was isolated as a mixture of enantiomers where the (2'S,6'S,7S) configuration was the major enantiomer. *H NMR (300 MHz, Chloroform-7) δ 9.40 (s, 1H), 9.07 (s, 1H), 7.48 (s, 1H), 6.60 (s, 1H), 3.87 (s, 2H), 3.69 (s, 1H), 3.36 (s, 1H), 2.61 (d, J = 7.0 Hz, 2H), 2.29 (d, J = 14.7 Hz, 1H), 2.20 - 1.97 (m, 5H), 1.86 (t, J = 13.3 Hz, 1H), 1.64 (s, 3H), 1.18 (s, 1H), 1.08 (s, 1H), 0.96 (s, 1H). LCMS m/z 355.28 [M+H]⁺. The site of N-acylation was tentatively assigned.

360

Compound 380

[l-[(2 'S, 6'S, 7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine] -2 'yl]cyclopropyl]methanol (380)

₃₇₂ C129 380

Step 1. 1-[(2'S,6'S, 7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-2'-yl]cyclopropanecarboxylic acid (C129)

To a mixture of methyl l-[(2'S,6'S,7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-2'-yl]cyclopropanecarboxylate 372 (82 mg, 0.22 mmol) in THF (500 pL) was added an aqueous (500 pL) solution of LiOH (26 mg, 1.1 mmol). The reaction mixture was heated to 50 °C for 2 hours, and additional THF (1 mL) was added to help dissolve the starting material. After heating for almost 3 days, the reaction was cooled to room température. The reaction mixture was then concentrated to remove the solvent and redissolved in DMSO. Purification by reversed-phase HPLC (Method: Waters XSelect CSH Cl8 OBD Prep Column; 30 x 150 mm, 5 micron. Gradient: acetonitrile in water with 5 mM hydrochloric acid) was isolated as l-[(2'S,6'S,7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-2'-yl]cyclopropanecarboxylic acid C129 (58 mg, 77% yield). Due to partial érosion of enantiomeric purity when preparing 372, C129 was isolated as a mixture of enantiomers where the (2'S,6'S,7S) configuration was the major enantiomer. *H NMR (300 MHz, DMSO-d6) δ 12.98 (s, 1H), 6.93 (s, 1H), 3.89 (t, J = 5.4 Hz, 2H), 3.48 (s, 2H), 2.58 (s, 3H), 2.22 - 1.84 (m, 4H), 1.29 (d, J = 6.3 Hz, 3H), 1.27 - 1.11 (m, 2H), 1.02 (s, 1H). LCMS m/z 342.27 [M+H]⁺.

Step 2. [1-[(2'S,6'S, 7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine]-2 ’-yl] cyclopropyl] methanol (380) l-[(2'S,6'S,7S)-2-chloro-6'-methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]2’-yl]cyclopropanecarboxylic acid C129 (15 mg, 0.044 mmol) was dissolved in THF (200 pL) in a vial. The vial was cooled to 0 °C under nitrogen. Borane tetrahydrofuran complex (175 pL of 1 M, 0.1750 mmol) was slowly added, and significant bubbling was observed. The mixture was allowed to slowly warm to room température. After 4 hours, the reaction was quenched with saturated NaHCCh solution and extracted with DCM (5x). The combined organic layer was dried

361 over Na2SÜ4, filtered, and concentrated in vacuo. Purification by reversed-phase HPLC (Method: Waters XSelect CSH C18 OBD Prep Column; 30 x 150 mm, 5 micron. Gradient: acetonitrile in water with 0.2% formic acid) afforded the product [l-[(2'S,6'S,7S)-2-chloro-6'methyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-2'-yl]cyclopropyl]methanol (formic acid sait) 380 (6.9 mg, 42% yield). Due to partial érosion of enantiomeric purity when preparing C129, 380 was isolated as a mixture of enantiomers where the (2'S,6'S,7S) configuration was the major enantiomer. ’H NMR (400 MHz, Methanol-A) δ 6.74 (s, 1H), 3.93 (t, J = 5.5 Hz, 2H), 3.86 (dd, J =11.7, 1.5 Hz, 1H), 3.66 - 3.47 (m, 1 H), 3.24 (d, J =11.7 Hz, 1H), 3.06 (dd,J=12.4, 2.6 Hz, 1H), 2.63 (t, J = 5.5 Hz, 2H), 2.27 (ddt, J = 16.7, 14.2, 2.8 Hz, 2H), 2.10 (dd, J = 14.6, 12.5 Hz, 1H), 1.85 (dd, J = 14.7, 12.2 Hz, 1H), 1.36 (d, J = 6.6 Hz, 3H), 0.93 - 0.79 (m, 1H), 0.79 - 0.67 (m, 1H), 0.56 (ddt, J = 17.2, 9.5, 5.4 Hz, 2H). LCMS m/z 328.28 [M+H]⁺.

Compound 381 (2 'R, 6'S, 7S)-2-chloro-2 ’-cyclopropyl-6'-(l-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] (381)

C130

381

Step 1. tert-butyl (2S,3R,6R)-6-cyclopropyl-2-(l-methyltriazol-4-yl)-4-oxo-piperidine-3carboxylate (Cl31)

To a solution of (3R)-3-(tert-butoxycarbonylamino)-3-cyclopropyl-propanoic acid C130 (500 mg, 2.181 mmol) in THF (4.5 mL) was added CDI (390 mg, 2.405 mmol), and the mixture

362 was stirred at room température for 3 hours. Bis[(3-tert-butoxy-3-oxo-propanoyl)oxy]magnesium C109 (449 mg, 1.310 mmol) was added. After 20 hours, the reaction was diluted with MBTE (10 mL) and 1 N HCl (3 mL). The organic layer was separated and washed with saturated aqueous sodium bicarbonate (3 mL) and brine (3 mL), dried with magnésium sulfate, fïltered, and concentrated to yield tert-butyl (5R)-5-(tert-butoxycarbonylamino)-5-cyclopropyl-3-oxopentanoate.

The crude material was dissolved in DCM (4 mL) and TFA (1 mL, 13 mmol) was added. After 1 h 30 min, the solution was azeotroped three times with DCM (4 mL).

The crude material from the second step was dissolved in DCM (4 mL) and 1methyltriazole-4-carbaldehyde (250 mg, 2.250 mmol) was added. After stirring over 2 days, the mixture was directly loaded onto a silica gel column for purification (Gradient: 0-10% MeOH in DCM). The product-containing fractions were pooled and concentrated to yield tert-butyl (2S,3R,6R)-6-cyclopropyl-2-(l-methyltriazol-4-yl)-4-oxo-piperidine-3-carboxylate C131 (432 mg, 62% yield). Ή NMR (400 MHz, Chloroform-d) δ 10.17 (s, 1H), 8.11 (s, 1H), 7.50 (s, 1H), 4.22 (s, 3H), 4.09 - 4.04 (m, 1H), 2.67 - 2.64 (m, 1H), 2.49 - 2.39 (m, 2H), 1.39 (s, 9H), 0.95 (dq, J = 8.1, 2.7 Hz, 1H), 0.57 (dd, J = 8.2, 1.5 Hz, 2H), 0.32 - 0.24 (m, 2H).

Step 2. (2 'R, 6'S, 7S)-2-chloro-2 '-cyclopropyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] (381)

To a mixture of tert-butyl (2S,3R,6R)-6-cyclopropyl-2-(l-methyltriazol-4-yl)-4-oxopiperidine-3-carboxylate C131 (25 mg, 0.078 mmol) in DCM (0.5 mL) was added MsOH (20 pL, 0.3082 mmol) and the reaction was refluxed. After 1 hour, 2-(5-chloro-3-thienyl)ethanol S2 (20 mg, 0.1230 mmol) was added. The reaction was stirred ovemight. The mixture was cooled to room température and diluted with additional DCM (2 mL) and saturated aqueous sodium bicarbonate (2 mL). The organic layer was separated and loaded onto a silica gel column for purification (Gradient: 0-10% MeOH in DCM). The product-containing fractions were found to be still under the purity threshold desired, so the combined fractions were concentrated and rediluted in DMSO. Purification by reversed-phase HPLC. Method: Waters XSelect CSH Cl8 OBD Prep Column; 30 x 150 mm, 5 micron. Gradient: acetonitrile in water with 0.1% trifluoroacetic acid. The product-containing fractions were diluted with DCM (1 mL) and saturated aqueous sodium bicarbonate (1 mL). The organic layer was dried to yield (2R,6'S,7S)2-chloro-2'-cyclopropyl-6'-(l-methyltriazol-4-yl)spnO[4,5-dihydrothieno[2,3-c]pyran-7,4'piperidine] 381 (5.3 mg, 18% yield). ‘H NMR (400 MHz, Chloroform-d) δ 7.45 (s, 1H), 6.58 (s, 1H), 4.35 (dd, J = 11.7, 2.6 Hz, 1H), 4.06 (s, 3H), 3.99 - 3.85 (m, 2H), 2.68 - 2.50 (m, 2H), 2.38 2.25 (m, 2H), 2.20 (dt, J = 13.6, 2.6 Hz, 1H), 1.80 (dd, J= 13.5, 11.7 Hz, 2H), 1.65 (dd, J= 13.6, 363

11.4 Hz, 1H), 0.79 (qt, J = 8.5, 4.9 Hz, 1H), 0.46 (tdd, J = 10.3, 8.3, 5.4 Hz, 2H), 0.23 - 0.12 (m, 2H). LCMS m/z 365.3 [M+H]⁺.

Compound 382 (2'R,4S,6'S, 7S)-2-chloro-2'-cyclopropyl-6'-(l-methyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3

c]pyran-7,4'-piperidine]-4-ol (382)

381

C134

Q^H OH

382

Cp*RhCI(R,R)-Tsdpen

364

Step 1. (2R,6S)-2-cyclopropyl-6-(l-methyltriazol-4-yl)piperidin-4-one (C132)

To a mixture of tert-butyl (2S,3R,6R)-6-cyclopropyl-2-(l-methyltriazol-4-yl)-4-oxopiperidine-3-carboxylate C131 (400 mg, 1.248 mmol) in chloroform (8 mL) was added 2,2,2trifluoroacetic acid (500 pL, 6.490 mmol) and the reaction was refluxed. After stirring ovemight, the mixture was cooled to room température, diluted with DCM (10 mL), and washed with saturated aqueous sodium bicarbonate (10 mL). The organic layer was dried with magnésium sulfate, filtered, and concentrated and then minimally diluted in DCM and loaded onto a silica gel column for purification (Gradient: 0-10% MeOH in DCM). The product-containing fractions were pooled and concentrated to yield (2R,6S)-2-cyclopropyl-6-(l-methyltriazol-4-yl)piperidin4-one C132 (90 mg, 33% yield). ’H NMR (400 MHz, Chloroform-d) δ 7.50 (s, 1H), 4.22 - 4.15 (m, 1H), 4.12 (s, 3H), 2.67 - 2.64 (m, 2H), 2.61 - 2.56 (m, 1H), 2.47 - 2.40 (m, 1H), 2.19 (ddd, J = 11.6, 8.7, 3.0 Hz, 1H), 0.95 (ddt, J = 13.2, 8.4, 4.1 Hz, 1H), 0.59 - 0.52 (m, 2H), 0.33 - 0.21 (m, 2H).

Step 2. (2'R,6'S, 7S)-2-chloro-2'-cyclopropyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] (381)

To a mixture of (2R,6S)-2-cyclopropyl-6-(l-methyltriazol-4-yl)piperidin-4-one C132 (45 mg, 0.20 mmol) and 2-(5-chloro-3-thienyl)ethanol (43 mg, 0.26 mmol) in DCM (1000 pL) was added MsOH (90 pL, 1.4 mmol) and the mixture was refluxed. After stirring ovemight, the mixture was cooled to room température and quenched with saturated aqueous sodium bicarbonate (1 mL). The layers were separated, and the organic layer was directly loaded onto a silica gel column for purification (Gradient: 0-10% MeOH in DCM). The product-containing fractions were pooled and concentrated to yield (2'R,6'S,7S)-2-chloro-2'-cyclopropyl-6'-(lmethyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] 381 (74 mg, 90% yield) as a yellow oil. LCMS m/z 365.15 [M+H]⁺.

Step 3. 1-[(2'R,6'S, 7S)-2-chloro-2'-cyclopropyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-1 '-yl]-2,2,2-trifluoro-ethanone (Cl33)

To (2^,R,6'S,7S)-2-chloro-2'-cyclopropyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] 381 (74 mg, 0.203 mmol) in DCM (1 mL) cooled to 0 °C was added DIPEA (50 pL, 0.2871 mmol), followed by TF AA (30 pL, 0.2158 mmol). After 35 minutes, the mixture was quenched with 1 N HCl (1 mL). The organic layer was separated and passed over a phase separator. The organic layer was loaded onto a silica gel column for purification (Gradient: 0-50% EtOAc in heptane). The product-containing fractions were pooled and concentrated to yield l-[(2'R,6'S,7S)-2-chloro-2'-cyclopropyl-6'-(l-methyltriazol-4yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C133 (51 365 mg, 50% yield) as a white solid. LCMS m/z 461.31 [M+H]⁺.

Step 4. (2R, 4S, 6S)-2 '-chloro-2-cyclopropyl-6-(l-methyltriazol-4-yl)-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one (C134)

A mixture ofl-[(2'R,6'S,7S)-2-chloro-2'-cyclopropyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C133 (51 mg, 0.102 mmol) in chlorobenzene (1 mL) was vacuum purged with nitrogen 5 times. At this time, 5,5dimethyl-l,3-dibromohydantoin (20 mg, 0.07 mmol) and 2-[(E)-(l-cyano-l-methyl-ethyl)azo]-2methyl-propanenitrile (1.5 mg, 0.009 mmol) were added, and the mixture was once again vacuum purged 5 times. The mixture was heated to 75 °C. After 15 minutes, the mixture was cooled to room température and mixed with saturated aqueous sodium bicarbonate (1 mL). The organic layer was separated, the aqueous layer was washed with ethyl acetate (1 mL), and the combined organic layer was dried with magnésium sulfate, filtered, and concentrated. The material was dried ovemight to remove residual solvent.

The crude foam was diluted with DMSO (1 mL) in a flame-dried flask and vacuum purged with nitrogen 5 times. The mixture was heated to 60 °C, triethylamine (75 pL, 0.54 mmol) was added, and the dark brown solution was heated further to 65 °C (internai température). After 75 minutes, the mixture was cooled to room température and diluted with water (5 mL) and ethyl acetate (5 mL). The aqueous layer was washed with ethyl acetate (2x5 mL), and then the combined organic layer was dried with sodium sulfate, filtered, and concentrated. The crude material was diluted with minimal DCM and loaded onto a silica gel column for purification (Gradient: 0-50% EtOAc in heptane). The product-containing fractions were pooled and concentrated to yield (2R,4S,6S)-2'-chloro-2-cyclopropyl-6-(l-methyltriazol-4yl)-l-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one C134 (16 mg, 32%) as a white solid. LCMS m/z 475.21 [M+H]⁺.

Step 5. 1-[(2'R,4S,6'S, 7S)-2-chloro-2'-cyclopropyl-4-hydroxy-6'-(l-methyltriazol-4yl)spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 '-piperidine]-1 '-yl]-2,2,2-trifluoro-ethanone (C135)

To a mixture of 5:2 diethylethanamine/formic acid (20 pL, 0.04763 mmol) in ACN (500 pL) was added a solution of N-[(lR,2R)-2-amino-l,2-diphenyl-ethyl]-4-methylbenzenesulfonamide (0.12 mg, 3.274E-4 mmol) and 1,2,3,4,5-pentamethylcyclopentane rhodium(2+) tetrachloride (0.1 mg, 1.592E-4 mmol) in ACN (50 pL). After 5 minutes, the solution was added to (2R,4S,6S)-2'-chloro-2-cyclopropyl-6-(l-methyltriazol-4-yl)-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4^,-one C134 (16 mg, 0.03203 mmol), and the reaction was stirred at -10 °C and warmed slowly ovemight. At this time, the mixture

366 was concentrated and directly loaded onto a silica gel column for purification (Gradient: 0-50% EtOAc in heptane). The product-containing fractions were pooled and concentrated to yield 1[(2'R,4S,6'S,7S)-2-chloro-2^,-cyclopropyl-4-hydroxy-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C135 (15 mg, 100%). LCMS m/z 477.24 [M+H]⁺.

Step 6. (2'R,4S,6'S, 7S)-2-chloro-2'-cyclopropyl-6'-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4’-piperidine]-4-ol (382) l-[(2'R,4S,6'S,7S)-2-chloro-2'-cyclopropyl-4-hydroxy-6^,-(l-methyltriazol-4-yl)spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C135 (15 mg, 0.03 mmol) was dissolved in MeOH (0.2 mL) and heated to 60 °C, at which point NaOH (50 pL of 6 M, 0.30 mmol) was added. After stirring for 7 hours, near full conversion was observed. The mixture was diluted with water (1 mL). Purification by reversed-phase HPLC (Method: Waters XSelect CSH Cl8 OBD Prep Column; 30 x 150 mm, 5 micron. Gradient: acetonitrile in water with 0.1% trifluoroacetic acid) afforded (2'R,4S,6'S,7S)-2-chloro-2'-cyclopropyl-6'-(lmethyltriazol-4-yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol (trifluoroacetate sait) 382 (11.7 mg, 74% yield over 2 steps) as a white solid. *H NMR (400 MHz, Methanol-d4) δ 8.03 (s, 1H), 6.95 (s, 1H), 4.49 (t, J = 3.4 Hz, 1H), 4.13 (s, 3H), 4.04 (dd, J = 12.2, 3.3 Hz, 1H), 3.86 (dd, J = 12.2, 3.7 Hz, 1H), 3.32 (d, J = 1.8 Hz, 1H), 2.99 (s, 1H), 2.64 - 2.50 (m, 2H), 2.28 (dd, J = 14.8, 12.6 Hz, 1H), 2.12 - 2.01 (m, 1H), 1.02 (dd, J = 8.7, 4.8 Hz, 1H), 0.74 (d, J = 8.1 Hz, 2H), 0.65 - 0.59 (m, 1H), 0.42 (s, 1H). LCMS m/z 381.28 [M+H]⁺.

Compound 383 (2 'R, 4S, 6'S, 7S)-2-chloro-2 ’-cyclopropyl-6'-(l-methyltriazol-4-yl)spiro[4,5-dihydrothieno [2,3c]pyran- 7,4 ’-piperidine]-4-ol (383)

³⁸³

To a mixture of (2R,6S)-2-cyclopropyl-6-(l-methyltriazol-4-yl)piperidin-4-one C132 (45 mg, 0.20 mmol) and 2-[5-(trifluoromethyl)-3-thienyl]ethanol S3 (52 mg, 0.27 mmol) in DCM

367 (1000 pL) was added MsOH (90 pL, 1.387 mmol) and the mixture was refluxed. After stirring ovemight, the mixture was cooled to room température and quenched with saturated aqueous sodium bicarbonate (1 mL). The layers were separated, and the organic layer was directly loaded onto a silica gel column for purification (Gradient: 0-10% MeOH in DCM). The productcontaining fractions were pooled and concentrated; however, UPLC analysis indicated presence of the intermediate. The product was dissolved in DMSO. Purification by reversed-phase HP LC (Method: Waters XSelect CSH Cl8 OBD Prep Column; 30 x 150 mm, 5 micron. Gradient: acetonitrile in water with 5 mM hydrochloric acid) afforded (2'R,6'S,7S)-2'-cyclopropyl-6'-(lmethyltriazol-4-yl)-2-(trifluoromethyl)spiro [4,5-dihydrothieno [2,3-c]pyran-7,4'-piperidine] (hydrochloride sait) 383 (18.7 mg, 21% yield) as a white solid. ’H NMR (400 MHz, Methanold4) δ 8.05 (s, 1H), 7.35 (s, 1H), 4.90 - 4.87 (m, 1H), 4.13 (s, 3H), 4.03 (t, J = 5.4 Hz, 2H), 2.99 (s, 1H), 2.78 (d, J = 2.4 Hz, 2H), 2.54 (d, J = 17.3 Hz, 2H), 2.46 - 2.37 (m, 1H), 2.15 - 2.05 (m, 1H), 1.03 (s, 1H), 0.74 (d, J = 8.2 Hz, 2H), 0.66 - 0.61 (m, 1H), 0.40 (s, 1H). LCMS m/z 399.3 [M+H]⁺.

Compound 384 (2'S,6'S, 7S)-2',4-dimethyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[5H-thieno[2,3c]pyran- 7,4 ’-piperidine]-4-ol (384)

O

S32 ³⁸⁴

To a mixture of (2S,4S,6S)-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2-trifluoroacetyl)-2'(trifluoromethyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one S32 (53 mg, 0.1036 mmol) in diethyl ether (1 mL) cooled to 0 °C was added bromo(methyl)magnesium (35 pL of 3.4 M, 0.1190 mmol). The mixture was stirred at 0 °C for 5 minutes, then quenched with saturated ammonium chloride (2 mL) and diluted with TBME (3 mL). The organic layer was washed with brine and dried over MgSCU, fïltered, and concentrated to a crude residue.

To the crude residue diluted in MeOH (1 mL) was added NaOH (50 pL of 6 M, 0.3000 mmol). The mixture was stirred at 60 °C for 2 hours. The reaction mixture was directly purified 368 by reversed-phase HPLC (Method: C18 Waters Sunfire column (30x150 mm, 5 micron).

Gradient: MeCN in H2O with 0.1% trifhioroacetic acid) to provide the product (2'S,6'S,7S)-2',4dimethyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[5H-thieno[2,3-c]pyran-7,4'piperidine]-4-ol (Trifluoroacetate sait) 384 (10.6 mg, 20%) as a clear glassy solid. LCMS m/z

403.27 [M+l]⁺.

Compound 385 (2S, 4S, 4 'S, 6S)-2'-chloro-4 ',5',5 '-trideuterio-2-methyl-6-(l -methyltriazol-4-yl)spiro[piperidine4,7'-thieno[2,3-c]pyran]-4'-ol (385)

Step 1. (2S, 4S, 6S)-2 '-chloro-5 5 '-dideuterio-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-cJpyran]-4'-one (C137)

369

To a 40 mL vial was added (2S,4S,6S)-2'-chloro-2-methyl-6-(l-methyltriazol-4-yl)-l(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4^,-one C136 (404 mg, 0.817 mmol), followed by D₂O (1.9 mL) and MeCN (5.6 mL). Then pyrrolidine (7 pL, 0.08 mmol) was added. Additional MeCN (1 mL) and dioxane (2 mL) were added, and the reaction mixture was heated to 75 °C. After 18 hours, the reaction mixed was washed with water and extracted with EtOAc (3x). The combined organic layer was washed with brine, dried over Na₂SÛ4, filtered, and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0 to 50% EtOAc in heptane) afforded (2S,4S,6S)-2'-chloro-5',5'-dideuterio-2-methyl-6-(lmethyltriazol-4-yl)-l-(2,2,2-trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one C137 (366 mg, 97%). LCMS m/z 451.15 [M+H]⁺. 97.5% D based on ‘H NMR.

Step 2. 1-[(2S, 4S, 4 'S, 6S)-2 '-chloro-4 ',5',5 '-trideuterio-4 '-hydroxy-2-methyl-6-(lmethyltriazol-4-yl)spiro[piperidine-4,7'-thieno[2,3-c]pyra.n]-l-yl]-2,2,2-trifluoro-ethanone (C138)

To a mixture of HCOOH-d4 (23 pL, 0.6095 mmol) and triethylamine (32 pL, 0.23 mmol) in DCM (500 pL) was added a solution of 1,2,3,4,5-pentamethylcyclopentane rhodium(2+) tetrachloride (0.195 mg, 3.104E-4 mmol) and N-[(lR,2R)-2-amino-l,2-diphenylethyl]-4-methyl-benzenesulfonamide (0.222 mg, 6.058E-4 mmol) in DCM (60 pL). After 5 minutes, (2S,4S,6S)-2'-chloro-5',5'-dideuterio-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one C137 (50 mg, 0.11 mmol) in DCM (500 pL) was added. The resulting orange reaction mixture was stirred at room température ovemight. The reaction was washed with saturated aqueous sodium bicarbonate, 1 N HCl, and brine, and the aqueous layers were extracted with EtOAc separately. The combined organic layer was dried over Na₂SO4, filtered, and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-50% EtOAc in heptane) afforded the product 1[(2S,4S,4'S,6S)-2'-chloro-4^,,5',5^,-trideuterio-4'-hydroxy-2-methyl-6-(l-methyltriazol-4yl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-l-yl]-2,2,2-trifluoro-ethanone C138 (45.8 mg, 93%). ‘H NMR (300 MHz, Chloroform-d) δ 7.59 (s, 1H), 6.84 (s, 1H), 5.53 (s, 1H), 4.68 - 4.39 (m, 1H), 4.13 - 4.03 (m, 3H), 3.10 (dd, J = 15.1, 7.4 Hz, 1H), 2.64 (ddd, J = 15.1, 8.1, 2.2 Hz, 2H), 2.12 (s, 1H), 2.08 (s, 1H), 1.44 - 1.25 (m, 3H). LCMS m/z 454.17 [M+H]⁺.

Step 3. (2S,4S, 4'S, 6S)-2'-chloro-4',5',5'-trideuterio-2-methyl-6-(l-methyltriazol-4yl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4’-ol (385)

To a solution of l-[(2S,4S,4'S,6S)-2'-chloro-4^,,5',5^,-trideuterio-4'-hydroxy-2-methyl-6-(lmethyltriazol-4-yl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-l-yl]-2,2,2-trifluoro-ethanone C138 (45.8 mg, 0.1008 mmol) in MeOH (800 pL) was treated with an aqueous solution of NaOH (170 370 pL of 6 Μ, 1.020 mmol) and heated to 60 °C. After 1 hour, the reaction vial was cooled to and quenched with saturated aqueous NaHCCh solution and extracted with EtOAc (4x). The combined organic layer was dried over sodium sulfate and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-12% MeOH in DCM) afforded the product (2S,4S,4'S,6S)-2^,-chloro-4',5',5'-trideuterio-2-methyl-6-(l-methyltriazol-4-yl)spiro[piperidine4,7'-thieno[2,3-c]pyran]-4'-ol 385 (24.3 mg, 61% yield). Ή NMR (400 MHz, Chloroform-d) δ 7.45 (s, 1H), 6.82 (s, 1H), 4.38 (dd, J = 11.7, 2.6 Hz, 1H), 4.05 (s, 3H), 3.47 (s, 1H), 3.37 (dqd, J = 12.7, 6.4, 2.5 Hz, 1H), 2.40 (dt, J = 13.9, 2.6 Hz, 1H), 2.03 (dt, J = 13.5, 2.6 Hz, 1H), 1.69 (dd, J = 13.9, 11.8 Hz, 1H), 1.50 (dd, J= 13.5, 11.3 Hz, 1H), 1.13 (d, J = 6.3 Hz, 3H). LCMS m/z 358.13 [M+H]⁺.

Préparation of Compound 476-1

TMSN₃

Toluene

- 90 °C

476-1

To a 500 mL three-necked RBF was added toluene (125 mL) and methyl propiolate (10.09 g, 1.0 eq.). TMSN3 (27.65 g, 2.0 eq.) was slowly added at room température. The mixture was heated to 85-90 °C for 24 hours. The solution was cooled to 0 °C and diluted with THF (200 mL). Solid NaNO₂ (10.76 g, 1.3 eq.) was added to the mixture. To this was added 2.4 M HCl (aqueous) (63 mL,1.3 eq.) dropwise while keeping the internai température at 0-10 °C. After addition, the mixture was stirred for 1 hour at 0-10 °C. The layers separated, and the aqueous layer was extracted with THF (2 x 200 mL). The combined organic extracts were washed with 3% NaHCCh (aqueous) (100 mL). The aqueous layer was extracted with THF (2 x 100 mL). The combined organic extracts were dried over Na₂SC>4, filtered, and concentrated under reduced pressure. The crude residue was washed with hexane (200 mL) and stirred for 1 hour at room température. The mixture was filtered, and the solid was washed with hexane (2x100 mL) to give a white powder (12.3 g, 81 % yield). Ή NMR (400 MHz, DMSO): δ 8.53 (s,lH),3.83 (s,3H).

371

Préparation of Compound 477-1a

476-1 ¹³cd₃i

N32CO3 THF °C

477-1 a

To a 500 mL three-necked round bottom flask was added THF (200 mL), 476-1 (14.8 g, 1.0 eq), and Na2CO₃ (18.6 g, 1.5 eq) followed by slow, portionwise addition of ¹³CD₃I (20.0 g, 1.2 eq.). The mixture was stirred for 48 hours at 25 °C. After reaction completion (TLC 1:1 Ethyl acetate: Hexane), the mixture was filtered through a Na2SO₄ pad. The cake was rinsed with DCM and THF. The filtrate was concentrated under vacuum followed by dissolution in DCM (145 mL) and washed with 5% Na2SO₃ (aqueous) (80 mL). The aqueous phase was extracted with DCM (4 x 80 mL). The combined organic extracts were concentrated and washed with hexane (6 x 300 mL). The solid was collected by filtration and dried to give 477-la as a white powder (8.4 g, 50.6% yield). Ή NMR (400 MHz, DMSO): δ 8.70 (d, J= 0.8 Hz, 1H), 3.82 (s, 3H), 3.32 (s, 3H).

Préparation of Compound 497a

477-1 a

DIBAL-H DCM

-70 °C d₃ ¹³c

497a

To a round bottom flask was added 477-la (10.0 g, 68.9 mmol) and 240 mL of anhydrous DCM. The reaction mixture was cooled to -70 °C. DIBAL (137 mL, IM in DCM, 137.8 mmol) was added by addition tunnel. The reaction mixture was stirred at -70 °C for 2 hours. Methanol (35 mL) was added at -70 °C by addition tunnel and stirred for 1 hour. Sodium tartrate (40%, 170 mL) was added slowly. The mixture was stirred at room température ovemight. The DCM layer was separated and the aqueous layer was extracted with DCM (4 x 150 mL). The combined organic extracts were concentrated, and hexanes (150 mL) was added to produce a white slurry, which was filtered and dried to give 497a (7.5 g, 94% yield) as a white solid. Ή NMR (400 MHz, CDC1₃) δ 10.14 (s, 1H) 8.09 (s, 1H).

372

Préparation of Compound 495a

ΟχΑ/) tBuOH

Γ DCC I I

OH OH OH O^/ MeCN * = ¹³C rt _495a

To a round bottom flask was added /BuOH (28.2 g, 380.7 mmol), [2-¹³C]malonic acid (20.0 g, 190.4 mmol) and MeCN (300 mL). DCC (43.2 g, 209.4 mmol) in MeCN (300 mL) was added at 5 °C in an ice-water bath. The white slurry was stirred at room température ovemight. The reaction mixture was filtered and washed with MeCN (2x30 mL). The filtrate was concentrated to give a yellow oil, which was purified by silica gel flash chromatography (100% DCM to 4% MeOH in DCM) to provide 495a (20.7 g 67 % yield) as a colorless oil. Ή NMR( 400 MHz, CDC1₃) δ 3.35 (d, J= 132 Hz, 2H), 1.49 (s, 9H).

Préparation of Compound 496a

To a solution of compound 495a (19.50 g, 121.0 mmol) in THF (150 mL) was added Mg(OtBu)? (11.46 g, 60.50 mmol) in portions. After each addition, a minor exothermic was observed, with the round bottom flask being warm to the touch (~30 °C). The mixture was stirred at room température ovemight. After 3 hours, the reaction was a clear, pale yellow solution. The mixture was concentrated via rotovap, and the residue was taken up in ether (50 mL) and concentrated again. This was repeated 3 times. The resulting solid was dried under vacuum to afford an off-white solid (21.20 g, 102% yield). In a separate flask, compound 479 (22.13 g, 108.9 mmol) in THF (230 mL) was treated with CDI (19.04 g, 117.4 mmol) in three portions. The reaction was stirred at room température under nitrogen and gas évolution was observed. After 3 hours, the previously prepared solid magnésium sait (21.20 g) was added and the reaction was stirred at room température ovemight. The reaction mixture was poured into 1 N HCl (270 mL) and extracted with EtOAc (2 x 200 mL). The combined organic extracts were washed with saturated sodium bicarbonate (200 mL) and brine (200 mL), dried over sodium sulfate, and concentrated via rotovap to afford a brown oil (crade, 35.5 g). This material was

373 taken up in THF (100 mL) and treated with 1 N NaOH (100 mL) and stirred at room température for 0.5 hour. This solution was diluted with water (300 mL) and extracted with EtOAc (3 x 300 mL). The combined organic extracts were washed with 0.5 HCl (100 mL) and brine (200 mL), dried over sodium sulfate, and concentrated via rotovap to give compound 496a (28.0 g, 85% yield) as a light yellow oil. ’H NMR (400 MHz, DMSO-d₆): δ 6.75 (d, J= 7.2 Hz, 1H), 3.83 (p, J = 7.2 Hz, 1H), 3.43 (d, J= 132 Hz, 2 H), 2.66-2.53 (m, 2H), 1.39 (s, 9H), 1.35 (s, 9H),1.00 (d, J = 6.8 Hz, 3H).

Préparation of Compound 498a

To a solution of compound 496a (28.0 g, 92.6 mmol) in DCM (110 mL) was added TFA (28.4 mL, 370.4 mmol) at room température. After 20 hours, TLC showed the reaction was completed. The mixture was concentrated via rotovap, and the residue brought up in DCM (100 mL) and concentrated again. This was repeated 3 times. The resulting solid was dried under vacuum to afford a pink solid (35.0 g). The solid was immediately dissolved in DCM (220 mL (sonicated to dissolve ail solids)) and treated with compound 497a (10.13 g, 88.0 mmol). The reaction was stirred at room température ovemight. TLC showed complété consumption of 497a and formation of the product. The reaction was quenched with saturated sodium bicarbonate (270 mL) and extracted with DCM (3 x 300 mL). The organic extracts were washed with brine (200 mL) and concentrated via rotovap to give crude solid (23.0 g). The crude solid was triturated in MTBE (70 mL) at room température for 1 hour. The white solid was collected via vacuum filtration, washed with cold MTBE (2 x 20 mL), and dried under high vacuum to afford compound 498a (14.1 g, 53% yield) as a white solid. ‘H NMR (400 MHz, CDC1₃): δ 7.46 (s, 1H), 4.50 (d, J= 10.8 Hz, 1H), 3.59 (dd, J= 11.2, 132 Hz, 1H), 3.21-3.16 (m, 1H) 2.55-2.50 (m, 1H), 2.22-2.16 (m, 1H), 2.00-1.90 (br, 1H), 1.37 (s, 9H), 1.27 (d, 7=6.4 Hz, 3H).

374

Préparations of Compounds 499a and 500a

MsOH, DCM ¹³cd₃

8.5:1 cis:trans 498a

¹³cd₃ >20:1 cis:trans 500a

To a solution of compound 498a (14.1, 47.1 mmol) in DCM (180 mL) was added MsOH (15.3 mL, 235.5 mmol). The reaction was heated to 40 °C ovemight and was monitored by TLC until the reaction was completed to give compound 499a. The reaction mixture was cooled to room température and quenched with saturated sodium bicarbonate solution, layers were separated, and the aqueous layer was extracted with DCM (5 x 120 mL). The combined organic extracts were concentrated via rotovap to afford compound 499a (crade, 9.15 g) as a pale yellow solid. The crade solid was triturated in MTBE (60 mL) for 3 hours, then filtered to give a white solid (8.30 g), which was further purified by silica gel flash chromatography (2-3% MeOH in DCM) to give pure compound 500a (6.80 g 72% yield). *H NMR (400 MHz, CDCh): δ 7.45 (s, 1H), 4.23 (m, 1H), 3.18-3.10 (m, 1H), 2.84-2.72 (m, 1H), 2.49-2.41 (m, 1H + 2 x 0.5H), 2.212.14 (m, 2 x 0.5H), 2.06-1.90 (br s 1H), 1.25 (d, J= 6.0 Hz, 3H).

375

Compound 386 (2S,4S,4S,6S)-2-methyl-6-(l-(l^I3C)methyltriazol-4-yl)-2’-(trifluoromethyl)spiro[(5^ViC)azinane4,7'-4,5-dihydrothieno[2,3-c]pyran]-4'-ol (386)

C139 C140

1. AIBN, NBS, MeCN 100WCFL lamp

2. Et₃N, DMSO

3. 4-acetamido-TEMPO KBr, NaOCI

Dichloro(pentamethylcyclopent adienyl)rhodium(lll) dimer

Formic acid-Et₃N (R.R)-TsDPEN

C143

386

Step 1. Synthesis of (2S,4S,6S)-2-methyl-6-(l-(methyl-^C-d3)-lH-l,2,3-triazol-4-yl)-2'(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-5-¹³C (C140)

A 500 mL 3-neck flask fitted with a magnetic stirrer, a heating mantle, a température probe, a water cooled reflux condenser, and a nitrogen inlet/outlet was charged with (2S,6S)-2methyl-6-(l-(methyl-¹³C-d3)-lH-l,2,3-triazol-4-yl)piperidin-4-one-5-¹³C C139 (10.4 g, 52.20 mmol) and dichloromethane (180 mL), and stirred for 5 minutes. 2-[5-(trifluoromethyl)-3thienyl]éthanol (12.5 g, 63.71 mmol) was added to the mixture, followed by methanesulfonic acid (24 mL, 369.8 mmol). The resulting reaction mixture was warmed to 40 °C, and stirred at this température for 5 days. The reaction mixture was cooled to 0 °C with an ice/water bath, and basified with 6 N NaOH solution until pH = 11.5. The reaction mixture was extracted with DCM (2 x 100 mL). The combined organic fraction was dried over MgSCU, filtered, and concentrated under reduced pressure. The crude residue was treated with MTBE (200 mL), aged for 1 hour, and then filtered through medium fritted tunnel, washed with MTBE (50 mL), and dried under

376 vacuum to afford one crop of the desired product in 12 g as a white solid. The mother liquor was concentrated under reduced pressure. Purification by silica gel chromatography (Gradient: 020% MeOH:DCM) afforded a second crop of the desired product in 5.7 g as a white solid. The two isolated crops of product were combined to give (2S,4S,6S)-2-methyl-6-(l-(methyl-¹³C-d3)lH-l,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]5-¹³C (17.7 g, 90%) C140 as a white solid. Ή NMR (400 MHz, Chloroform-d) δ 7.44 (t, J = 0.7 Hz, 1H), 7.12 (q, J = 1.2 Hz, 1H), 4.44 (dt, J = 11.8, 2.8 Hz, 1H), 4.04 - 3.89 (m, 2H), 3.33 (dtd, J = 12.7, 6.4, 2.5 Hz, 1H), 2.79 - 2.61 (m, 2H), 2.38 (ddt, J = 131.3, 13.5, 2.6 Hz, 1H), 2.14 2.04 (m, 1H), 2.05 - 1.57 (m, 1H), 1.49 (dd, J = 13.7, 11.3 Hz, 1H), 1.13 (d, J = 6.3 Hz, 3H). ¹⁹F NMR (376 MHz, Chloroform-d) δ -55.30. LCMS m/z 378.07 [M+l]⁺.

Step 2. Synthesis of2,2,2-trifluoro-l-((2S,4S,6S)-2-methyl-6-(l-(methyl-^I3C-d3)-lH-l,2,3triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-1-yl5-^I3C)ethan-l-one (C141)

A 500 mL 3-neck flask fitted with a magnetic stirrer, a température probe, and a nitrogen inlet/outlet was charged with (2S,4S,6S)-2- methyl-6-(l-(l¹³C)methyltriazol-4-yl)-2'(trifluoromethyl)spiro[(5¹³C)azinane-4,7'-4,5-dihydrothieno[2,3-c]pyran] (17.5 g, 46.37 mmol) and DCM (200 mL), stirred for 5 minutes, and then cooled to 0 °C with an ice/water bath. To the mixture, N-ethyl-N-isopropyl-propan-2-amine (13 mL, 74.63 mmol) was added, followed by (2,2,2-trifluoroacetyl) 2,2,2-trifluoroacetate (7.5 mL, 53.96 mmol). The resulting reaction mixture was stirred at this température for 1 hour. The reaction mixture was quenched with a saturated NaHCCh solution (100 mL). The layers were separated, and organic layer was washed with 2 M aqueous HCl (2 x 60 mL), water (100 mL), brine (100 mL), then dried over MgSO₄, filtered and concentrated. Purification by silica gel chromatography (0-100% EtOAc:Heptane) provided 2,2,2-trifluoro-l-((2S,4S,6S)-2-methyl-6-(l-(methyl-¹³C-d3)-lH-l,2,3-triazol-4-yl)-2'(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-l-yl-5-¹³C)ethan-l-one C141 (21 g, 96%) as a white foam. Ή NMR (400 MHz, Chloroform-d) δ 7.59 (s, 1H), 7.12 (d, J = 1.3 Hz, 1H), 5.60 (s, 1H), 4.52 - 4.32 (m, 1H), 3.91 (t, J = 5.5 Hz, 2H), 3.24 (ddd, J = 130.5, 14.8, 7.4 Hz, 1H), 2.88 - 2.59 (m, 2H), 2.55 - 2.38 (m, 1H), 2.07 (d, J = 21.9 Hz, 1H), 1.49 -1.00 (m, 3H). ¹⁹F NMR (376 MHz, Chloroform-d) δ -55.41, -68.97. LCMS m/z 474.02 [M+l]⁺.

Step 3. Synthesis of (2S,4S,6S)-2-methyl-6-(l-(methyl-¹³C-d3)-lH-l,2,3-triazol-4-yl)-l-(2,2,2trifluoroacetyl)-2'-(trifluoromethyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'(5'H)-one-5¹³ C (C142)

First Step - Photochemical Bromination: AIL flask fitted with a magnetic stirrer, a 100 W CFL light source, and a nitrogen inlet/outlet was charged with 2,2,2-trifluoro-l-((2S,4S,6S)-2377 methyl-6-( 1 -(methyl- ¹³C-d3)-1 H-1,2,3 -triazol-4-yl)-2'-(trifluoromethyl)-4',5'dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-l-yl-5-^I3C)ethan-l-one C141 (21 g, 44.36 mmol) and acetonitrile (400 mL). The resulting reaction mixture was degassed with a stream of nitrogen via gas dispersion tube for 15 minutes. To the mixture, N-bromosuccinimide (10.2 g, 57.31 mmol) was added, followed by AIBN (200 mg, 1.218 mmol). The resulting reaction mixture was stirred under 100 W CFL irradiation for 4 hours. The reaction mixture was quenched with aqueous 10 wt% sodium bisulfite (200 mL), stirred for 10 minutes, and then MTBE (300 mL) was added. The organic phase was separated, washed with saturated aqueous sodium bicarbonate (200 mL) and brine (200 mL), dried over MgSCU, filtered, and concentrated to afford l-((2S,4S,6S)-4'-bromo-2-methyl-6-(l-(methyl-¹³C-d3)-lH-l,2,3-triazol-4-yl)-2'(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-l-yl-5-¹³C)-2,2,2trifluoroethan-l-one (25 g, 102%) as a tan foam. This crude material was taken into the next step without further purification. LCMS m/z 553.94 [M+l]⁺

Second Step - Kornblum Oxidation: A mixture ofl-((2S,4S,6S)-4'-bromo-2-methyl-6-(l(methyl-¹³C-d3)-lH-l,2,3-triazol-4-yl)-2^,-(trifluoromethyl)-4',5^,-dihydrospiro[piperidine-4,7'thieno[2,3-c]pyran]-l-yl-5-¹³C)-2,2,2-trifluoroethan-l-one (25 g, 102%) and dimethylsulfoxide (200 mL) was stirred for 5 minutes. To the reaction mixture, Et₃N (45 mL, 322.9 mmol) was added. The resulting reaction mixture was warmed to 75 °C and stirred for 3 hours. The reaction mixture was cooled to room température, partitioned between MTBE (~400 mL), saturated aqueous NaHCCh solution (~200 mL) and water (~400 mL), and stirred for 10 minutes. The organic phase was separated, and the aqueous layer was extracted with MTBE (2 x 200 mL). The combined organic phase was washed with saturated NaHCCh solution (~200 mL), dried over MgSÜ4, filtered, and concentrated to yield a product consisting of a mixture of the ketone (2S,4S,6S)-2-methyl-6-(l-(methyl-¹³C-d3)-lH-l,2,3-triazol-4-yl)-l-(2,2,2-trifluoroacetyl)-2'(trifluoromethyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'(5'H)-one-5-^I3C C142 and alcohol 2,2,2-trifluoro-l-((2S,4S,6S)-4'-hydroxy-2-methyl-6-(l-(methyl-¹³C-d3)-lH-l,2,3-triazol-4-yl)2'-(trifluoromethyl)-4^,,5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-l-yl-5-¹³C)ethan-lone (21 g, 97%) in a 5:1 ratio as a tan color foam. This crude material of ketone and alcohol (5:1) was taken to the following step without further purification. LCMS m/z 487.95 [M+l]⁺.

Third Step - Oxidation: A solution of the crude material from the second step (21 g, 42.91 mmol) (mixture of ketone and alcohol, ~5:1 ratio) and DCM (200 mL) was stirred for 5 minutes and then cooled to 0 °C. To the reaction mixture, solid NaHCO₃ (2.4 g, 28.57 mmol) was added, followed by KBr (1.5 g, 12.60 mmol) in water (25 mL). To the reaction mixture, 4acetamido-TEMPO (480 mg, 2.250 mmol) was added, followed by NaOCl (30 mL of 12 %

378 (w/w), 58.32 mmol) very slowly over 30 minutes while maintaining an internai température below 7 °C. The resulting reaction mixture was stirred for 1 hour. The reaction mixture was quenched with aqueous 1 M Na₂S₂Ü3 (100 mL). The organic phase was separated, washed with saturated aqueous NaHCCh solution (100 mL) and brine (100 mL), dried over MgSCU, and then filtered and concentrated. Purification by silica gel chromatography (Gradient: 0-100% EtOAc:Heptane) yielded (2S,4S,6S)-2-methyl-6-(l-(methyl-¹³C-d3)-lH-l,2,3-triazol-4-yl)-l(2,2,2-trifhioroacetyl)-2'-(trifluoromethyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'(5'H)-one5-¹³C C142 (14 g, 66%) as a white solid. ‘H NMR (400 MHz, Chloroform-d) δ 7.73 (q, J = 1.2 Hz, 1H), 7.62 (s, 1H), 5.65 (s, 1H), 4.47 (p, J = 7.1 Hz, 1H), 4.36 (d, J = 4.4 Hz, 2H), 3.40 (ddd, J = 131.1, 15.0, 5.8 Hz, 1H), 2.99 (dd, J = 15.0, 8.5 Hz, 1H), 2.65 (dd, J = 15.1, 8.5 Hz, 1H), 2.23 (d, J = 23.2 Hz, 1H), 1.26 (s, 3H). ^I9F NMR (376 MHz, Chloroform-d) δ -55.96, -68.87. LCMS m/z 488.01 [M+l]⁺

Step 4. Synthesis of2,2,2-trifluoro-l-((2S,4S,4'S,6S)-4’-hydroxy-2-methyl-6-(l-(methyl-^I3CdiflH-l, 2,3-triazol-4-yl)-2 '-(trifluoromethyl)-4 5 '-dihydrospiro[piperidine-4,7'-thieno[2,3c]pyran]-l-yl-5-¹³C)ethan-l-one (C143)

A solution of dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (72 mg, 0.1146 mmol) and (R,R)-TsDPEN (90 mg, 0.2456 mmol) was prepared in MeCN (120 mL) and then stirred for 1 hour at room température. Formic acid-Et3N (15.8 mL, 237.0 mmol, a 5:2 commercial solution) was added in one portion, and the reaction was then cooled to -15 °C.

In as separate flask, (2S,4S,6S)-2-methyl-6-(l-(methyl-¹³C-d3)-lH-l,2,3-triazol-4-yl)-l(2,2,2-trifluoroacetyl)-2^,-(trifluoromethyl)spiro[piperidine-4,7^,-thieno[2,3-c]pyran]-4^,(5'H)-one5-^I3C C142 (14 g, 28.44 mmol) in ACN (120 mL) was cooled to -15 °C, and then added to the previously prepared solution, while carefully maintaining the internai température between 17°C and-20 °C.

The resulting reaction mixture was warmed to -10 °C and stirred at this température for 6 hours. The reaction mixture was quenched with a saturated NaHCCh solution (200 mL) and stirred at room température for 6 hours. The reaction mixture was then extracted with MTBE (2 x 200 mL). The combined organic phase was washed with 1 N HCl (200mL) and brine (200mL), dried over sodium sulfate, filtered, and concentrated. The residue was dissolved in DCM (~200 mL), treated with 3-mercaptopropyl ethyl sulfide silica (SPM32f métal scavenging resin) (6 g), stirred for 2 hours at room température, and then filtered and washed with DCM (60 mL). The combined filtrate was concentrated. Purification by silica gel chromatography (Gradient: 0-100% EtOAc:Heptane) yielded 2,2,2-trifluoro-l-((2S,4S,4'S,6S)-4'-hydroxy-2-methyl-6-(l-(methyl¹³C-d3)-lH-l,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3379

c]pyran]-l-yl-5-¹³C)ethan-l-one C143 (13 g, 93%) as a white foam. Ή NMR (400 MHz, Chloroform-d) δ 7.60 (s, 1H), 7.38 (q, J = 1.2 Hz, 1H), 5.57 (s, 1H), 4.58 (dt, J = 9.1, 3.3 Hz, 1H), 4.50 (s, 1H), 3.99 (dd, J = 12.4, 3.2 Hz, 1H), 3.89 (dd, J = 12.4, 3.4 Hz, 1H), 3.17 (ddd, J = 131.1, 15.2, 7.0 Hz, 1H), 2.68 (ddd, J = 132.9, 15.2, 8.3 Hz, 2H), 2.20 (s, 1H), 2.10 (d, J = 9.1 Hz, 1H), 1.33 (s, 3H). ¹⁹F NMR (376 MHz, Chloroform-d) δ -55.56, -68.96. LCMS m/z 490.05 [M+l]⁺.

Step 5. Synthesis of (2S,4S,4'S,6S)-2-methyl-6-(l-(methyl-¹³C-d3)-lH-l,2,3-triazol-4-yl)-2'(trifluoromethyl)-4',5'-dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-5-¹³C-4'-ol (386)

A solution of 2,2,2-trifluoro-l-((2S,4S,4'S,6S)-4'-hydroxy-2-methyl-6-(l-(methyl-¹³Cd3)-lH-l,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4^,,5'-dihydrospiro[piperidine-4,7'-thieno[2,3c]pyran]-l-yl-5-¹³C)ethan-l-one C143 (13 g, 26.56 mmol) and MeOH (120 mL) was stirred for 5 minutes. To the reaction mixture, aqueous NaOH (45 mL of 6 M, 270.0 mmol) was added in one portion. The resulting reaction mixture was warmed to 60 °C and stirred at this température for 1 hour. The reaction mixture was cooled to room température, then partitioned between cold water (100 mL) and MTBE (200 mL) and stirred for 20 minutes. The organic phase was then separated. The aqueous phase was extracted with MTBE (2 x 100 mL). The combined organic phases were washed with cold water (60 mL), brine (100 mL), dried over MgSCU, filtered, and concentrated. The residue was dried under vacuum oven at 80 °C for 14 hours to afford (2S,4S,4^,S,6S)-2-methyl-6-(l-(methyl-^I3C-d3)-lH-l,2,3-triazol-4-yl)-2'-(trifluoromethyl)-4',5'dihydrospiro[piperidine-4,7'-thieno[2,3-c]pyran]-5-¹³C-4'-ol 386 (9.5 g, 90%) as a white solid. ’H NMR (400 MHz, Methanol-d4) δ 7.83 (t, J = 0.7 Hz, 1H), 7.46 (q, J = 1.2 Hz, 1H), 4.58 (t, J = 3.9 Hz, 1H), 4.35 (dt, J = 11.8, 2.9 Hz, 1H), 4.08 (dd, J = 12.2, 3.6 Hz, 1H), 3.86 (dd, J = 12.2, 4.2 Hz, 1H), 3.35 (ddd, J = 11.5, 6.3, 3.2 Hz, 1H), 2.64 - 2.17 (m, 2H), 1.78 (ddd, J = 127.7, 13.8, 11.8 Hz, 1H), 1.53 (dd, J= 13,6, 11.4 Hz, 1H), 1.17 (d, J = 6.4 Hz, 3H). ¹⁹F NMR (376 MHz, Methanol-d4) δ -56.94. LCMS m/z 394.1 [M+l]⁺.

380

Préparation of S63 tert-butyl (2S,3R, 6R)-2-(l-methyl-lH-1,2,3-triazol-4-yl)-4-oxo-6-phenylpiperidine-3-carboxylate

Step 1. To a solution of (3R)-3-(tert-butoxycarbonylamino)-3-phenyl-propanoic acid C144 (500 mg, 1.885 mmol) in THF (4.5 mL) was added CDI (340 mg, 2.097 mmol), and the mixture was stirred at room température for 2.5 hours. Bis[(3-tert-butoxy-3-oxopropanoyl)oxy]magnesium C109 (390 mg, 1.138 mmol) was then added, and stirring continued at room température for 20 hours. The reaction was diluted with TBME (10 mL) and 1 N HCl (3 mL). The organic layer was separated and washed with saturated sodium bicarbonate (3 mL), brine (3 mL). The organic layer was dried with magnésium sulfate, filtered, and concentrated to yield terZ-butyl (2S,3R,6R)-2-(l-methyl-lH-l,2,3-triazol-4-yl)-4-oxo-6-phenylpiperidine-3carboxylate S63 as a white crystalline solid, which was immediately carried forward to Step 2.

Step 2. To the white crystalline solid from the first step dissolved in DCM (4 mL) was added TFA (900 pL, 11.68 mmol) The mixture was stirred for 1 hour, and then the mixture was concentrated and azeotroped by DCM (3x4 mL) to provide a crude mixture of ZerAbutyl (5R)5-amino-3-oxo-5-phenyl-pentanoate that was directly carried forward to Step 3.

Step 3. To the crude mixture from step 2 dissolved in DCM (4 mL) was added 1methyltriazole-4-carbaldehyde S17 (225 mg, 2.025 mmol) and the mixture was stirred at room température for 2 hours. The mixture was quenched with saturated sodium bicarbonate (2 mL). 6 N NaOH was added to adjust the pH to >9. The organic layer was separated and concentrated. Purification by silica gel chromatography (Gradient: 0-10% MeOH:DCM) yielded tert-butyl (2S,3R,6R)-2-(l-methyltriazol-4-yl)-4-oxo-6-phenylpiperidine-3-carboxylate S63 (347 mg, 52%). This was observed as a mixture of keto and enol tautomers. LCMS m/z 357.23 [M+H]⁺

381

Préparation of S64 l-[(2'S, 6'R, 7S)-2-chloro-2 '-(l-methyltriazol-4-yl)-6'-phenyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-1 ’-yl] -2,2,2-trifluoro-ethanone (S64)

S63

Step 1. Synthesis of (2S,6R)-2-(l-methyltriazol-4-yl)-6-phenyl-piperidin-4-one (C145)

To a mixture of tert-butyl (2S,3R,6R)-2-(l-methyltriazol-4-yl)-4-oxo-6-phenylpiperidine-3-carboxylate S63 (320 mg, 0.8978 mmol) dissolved in DCM (5 mL) was added MsOH (300 pL, 4.623 mmol). The mixture was refluxed for 1 hour, then cooled to room température and quenched with saturated sodium bicarbonate (5 mL). The organic layer was separated, and the aqueous layer was extracted with additional DCM (3x5 mL). The combined organic layer was dried with MgSO₄, filtered, and concentrated. Purification by silica gel chromatography (0-10% MeOH:DCM) yielded (2S,6R)-2-(l-methyltriazol-4-yl)-6-phenylpiperidin-4-one C145 (98 mg, 43%), as a yellow oil. *H NMR (400 MHz, Chloroform-d) δ 7.53 (s, 1H), 7.46 - 7.42 (m, 2H), 7.40 - 7.30 (m, 3H), 4.41 (dd, J = 9.8, 5.2 Hz, 1H), 4.18-4.14 (m, 1H), 4.13 (s, 3H), 2.84 - 2.72 (m, 2H), 2.65 (d, J = 7.6 Hz, 2H).

Step 2. Synthesis of (2'S,6'R,7S)-2-chloro-2'-(l-methyltriazol-4-yl)-6'-phenyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] (C146)

To a mixture of (2S,6R)-2-(l-methyltriazol-4-yl)-6-phenyl-piperidin-4-one C145 (49 mg, 0.1912 mmol) and 2-(5-chloro-3-thienyl)ethanol S2 (40 mg, 0.2459 mmol) dissolved in DCM 382 (1000 pL) was added MsOH (90 pL, 1.387 mmol). The mixture was refluxed ovemight and then cooled to room température and quenched with saturated sodium bicarbonate (1 mL). The organic layer was directly purified by silica gel chromatography (Gradient: 0-10% MeOH:DCM) to yield (2'S,6'R,7S)-2-chloro-2^,-(l-methyltriazol-4-yl)-6'-phenyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] C146 (46 mg, 57%) as a white crystalline solid. *H NMR (400 MHz, Chloroform-d) δ 7.48 (s, 1H), 7.45 - 7.38 (m, 2H), 7.36 - 7.29 (m, 2H), 7.24 (td, J = 5.3, 4.8, 2.4 Hz, 1H), 6.58 (s, 1H), 4.58 (dd, J = 11.7, 2.6 Hz, 1H), 4.32 (dd, J = 11.6, 2.5 Hz, 1H), 4.06 (s, 3H), 4.02 (t, J = 5.5 Hz, 2H), 2.72 - 2.54 (m, 2H), 2.44 (dt, J = 13.6, 2.6 Hz, 1H), 2.22 (dt, J = 13.6, 2.6 Hz, 1H), 1.86 (ddd, J = 21.7, 13.6, 11.7 Hz, 3H). LCMS m/z 401.15 [M+l]⁺

Step 3. Synthesis of l-[(2'S,6'R,7S)-2-chloro-2'-(l-methyltriazol-4-yl)-6'-phenyl-spiro[4,5dihydrothieno[2,3-c]pyran- 7,4 ’-piperidine]-1 '-yl]-2,2,2-trifluoro-ethanone (S64)

A solution of (2'S,6'R,7S)-2-chloro-2'-(l-methyltriazol-4-yl)-6'-phenyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine] C146 dissolved in DCM (1 mL) was cooled to 0 °C. DIPEA (25 pL, 0.1435 mmol) was added, followed by TF AA (20 pL, 0.1439 mmol). The mixture was stirred for 20 min, and then quenched with 1 N HCl (1 mL). The organic layer was passed through a phase separator. Purification by silica gel chromatography (0-50% EtOAc:Heptane) yielded l-[(2'S,6'R,7S)-2-chloro-2'-(l-methyltriazol-4-yl)-6^,-phenyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone S64 (52 mg, 52%) as a clear oil. LCMS m/z 497.26 [M+l]⁺

383

Compound 387 (2 'S, 4S, 6'R, 7S)-2-chloro.-2 '-(l -methyltriazol-4-yl)-6'-phenyl-spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine]-4-ol (387)

S64

i) DBDMH, AIBN Et₃N ii) Et₃N, DMSO

Dichloro(pentamethylcyclopent adienyl)rhodium(III) dimer

C147

Formic acid-Et₃N (R.R)-TsDPEN

C148

Step 1. Synthesis of (2S,4S,6R)-2'-chloro-2-(l-methyltriazol-4-yl)-6-phenyl-1-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one (C147)

To a solution ofl-[(2'S,6^,R,7S)-2-chloro-2'-(l-methyltriazol-4-yl)-6^l-phenyl-spiro[4,5dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone S64 (52 mg, 0.09901 mmol) in chlorobenzene (1 mL) under nitrogen atmosphère was added 5,5-dimethyl-l,3dibromohydantoin (20 mg, 0.06995 mmol) and 2-[(E)-(l-cyano-l-methyl-ethyl)azo]-2-methylpropanenitrile (1.5 mg, 0.009135 mmol). The mixture was heated to 75 °C for 15 minutes. The mixture was cooled to room température and quenched with saturated sodium bicarbonate (1 mL). The organic layer was separated, dried with MgSCh, filtered, and concentrated to provide a crude foam.

The crude foam was dissolved in DMSO (1 mL) under nitrogen atmosphère. The mixture was heated to 60 °C and triethylamine (75 pL, 0.5381 mmol) was added. The dark brown solution was heated to 65 °C and stirred for 75 minutes. The mixture was cooled to room température, and diluted with ethyl acetate (5 mL) and water (5 mL). The aqueous layer was washed with additional ethyl acetate (2x5 mL). The combined organic layers were dried with Na2SÜ4, filtered, and concentrated. Purification by silica gel chromatography (Gradient: 0-50% EtOAc:Heptane) yielded (2S,4S,6R)-2'-chloro-2-(l-methyltriazol-4-yl)-6-phenyl-1-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one C147 (21 mg, 39%) as a pale

384 yellow solid. LCMS m/z 511.21 [M+l]⁺

Step 2. Synthesis of l-[(2'S,4S,6'R,7S)-2-chloro-4-hydroxy-2'-(l-methyltriazol-4-yl)-6'phenyl-spiro[4,5-dihydrothieno[2,3-c]pyran- 7,4 '-piperidine]-1 ’-yl] -2,2,2-trifluoro-ethanone (C148)

To a mixture of 5:2 formic acid-triethylamine complex (20 pL, 0.04763 mmol) in MeCN (500 pL) was added a solution of N-[(lR,2R)-2-amino-l,2-diphenyl-ethyl]-4-methylbenzenesulfonamide (0.12 mg, 3.274E⁴ mmol) and Dichloro(pentamethylcyclopentadienyl)rhodium(III) dimer (0.1 mg, 1.592E'⁴ mmol) dissolved in MeCN (50 pL). After 5 minutes, the resulting solution was added to a cooled solution at -10°C of (2S,4S,6R)-2'-chloro-2-(l-methyltriazol-4-yl)-6-phenyl-l-(2,2,2trifluoroacetyl)spiro[piperidine-4,7'-thieno[2,3-c]pyran]-4'-one C147 (21 mg, 0.03908 mmol) in MeCN (500 pL). The reaction mixture was warmed slowly to room température and stirred ovemight. The mixture was concentrated and purified by silica gel chromatography (Gradient: 050% EtOAc:heptane) to yield l-[(2'S,4S,6'R,7S)-2-chloro-4-hydroxy-2'-(l-methyltriazol-4-yl)6'-phenyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C148 as a white solid. LCMS m/z 514.94 [M+l]⁺

Step 3. Synthesis of (2'S,4S,6'R,7S)-2-chloro-2'-(l-methyltriazol-4-yl)-6'-phenyl-spiro[4,5dihydrothieno[2,3-c]pyran- 7,4 '-piperidine]-4-ol (387)

A solution of l-[(2^,S,4S,6'R,7S)-2-chloro-4-hydroxy-2'-(l-methyltriazol-4-yl)-6'-phenylspiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-r-yl]-2,2,2-trifluoro-ethanone C148 dissolved in MeOH (250 pL) was heated to 60 °C. NaOH (75 pL of 6 M, 0.4500 mmol) was then added, and reflux continued for 3 hours. The mixture was diluted with MTBE (3 mL) and saturated ammonium chloride (3 mL). The organic layer was separated, and the aqueous layer was extracted with additional MTBE (2x3 mL). The combined organic layer was passed over a phase separator and concentrated to yield (2'S,4S,6'R,7S)-2-chloro-2'-(l-methyltriazol-4-yl)-6'phenyl-spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine]-4-ol 387 (13.8 mg, 82%) as a white solid. 'H NMR (400 MHz, Methanol-d4) δ 7.88 (d, J = 3.2 Hz, 1H), 7.48 - 7.41 (m, 2H), 7.37 7.30 (m, 2H), 7.30 - 7.21 (m, 1H), 6.88 (s, 1H), 4.60 - 4.47 (m, 2H), 4.37 (dd, J = 11.8, 2.6 Hz, 1H), 4.12 (dt, J = 11.5, 3.4 Hz, 1H), 4.07 (s, 3H), 3.93 - 3.84 (m, 1H), 2.49 (dt, J = 13.9, 2.6 Hz, 1H), 2.35 -2.22 (m, 1H), 1.89 (ddd, J =13.8, 11.8, 1.5 Hz, 2H). LCMS m/z 417.26 [M+l]⁺385

Préparation of S65

2-(difluoromethyl)-6-(l-methyltriazol-4-yl)piperidin-4-one (S65)

O

!

[CIS]

To a solution of 4-amino-5,5-difluoro-pentan-2-one (Hydrochloride sait) C149 (250 mg, 1.440 mmol) in éthanol (15 mL) cooled to 0 °C was added l-methyltriazole-4-carbaldehyde S17 (175 mg, 1.512 mmol), L-proline (35 mg, 0.3040 mmol), and EtsN (210 pL, 1.507 mmol). The mixture was warmed to room température and 72 hours. The mixture was concentrated, and dissolved in DCM (10 mL) and saturated sodium bicarbonate (5 mL). The aqueous layer was extracted with additional DCM (2x10 mL), and the combined organic layer was concentrated. Purification by silica gel chromatography (Gradient: 0-10% MeOH:DCM) yielded 2(difluoromethyl)-6-(l-methyltriazol-4-yl)piperidin-4-one S65 (303 mg, 91%) as a yellow oil. 'H NMR (400 MHz, Chloroform-d) δ 7.51 (s, 1H), 5.78 (tdd, J = 56.0, 16.1, 4.3 Hz, 1H), 4.30 (dt, J = 9.8, 4.7 Hz, 1H), 4.13 (s, 3H), 3.51 (d, J = 5.0 Hz, 1H), 3.41 (d, J = 18.7 Hz, 1H), 2.79 - 2.69 (m, 1H), 2.60 (s, 1H), 2.43 (dd, J = 14.4, 11.9 Hz, 1H), 2.30 (s, 1H). LCMS m/z 231.18 [M+H]⁺. The product was isolated as a mixture of 4:1 dr.

Compound 388

2-chloro-2 ’-(difluoromethyl)-6'-(l -methyltriazol-4-yl)spiro[4,5-dihydrothienp[2,3-c]pyran- 7,4'piperidine] (388)

S65 388

[CIS] [CHF₂ and Metriazole are CIS]

386

To a solution of 2-(difluoromethyl)-6-(l-methyltriazol-4-yl)piperidin-4-one S65 (140 mg, 0.6081 mmol) in DCM (3 mL) was added 2-(5-chloro-3-thienyl)ethanol S2 (100 pL, 0.8085 mmol) followed by MsOH (200 pL, 3.082 mmol). The mixture was refluxed ovemight. The mixture was cooled to room température and quenched with saturated sodium bicarbonate. The organic layer was separated and concentrated. Purification by silica gel chromatography (Gradient: 0-10% MeOH:DCM) yielded 2-chloro-2'-(difluoromethyl)-6'-(l-methyltriazol-4yl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] 388 (139 mg, 53%) as a yellow oil. *H NMR (400 MHz, Chloroform-d) δ 7.47 (s, 1H), 6.62 (s, 1H), 5.66 (td, J = 56.5, 5.2 Hz, 1H), 4.48 (dd, J = 11.8, 2.7 Hz, 1H), 4.10 (d, J = 1.5 Hz, 3H), 3.98 (t, J = 5.5 Hz, 2H), 3.64 - 3.51 (m, 1H), 2.65 (td, J = 5.5, 3.1 Hz, 2H), 2.41 - 2.36 (m, 1H), 2.24 - 2.16 (m, 1H), 2.13 (dd, J = 14.8, 6.6 Hz, 1H), 1.84 (dd, J = 13.6, 11.8 Hz, 1H). LCMS m/z 375.14 [M+l]⁺.

Compound 389 '-(difluoromethyl)-6'-(l -methyltriazol-4-yl)-2-(trifluoromethyl)spiro[4,5-dihydrothieno[2,3c]pyran-7,4'-piperidine] (389)

S65 ³⁸⁹

[CIS] [CHF₂ and Metriazole are CIS]

To a solution of 2-(difluoromethyl)-6-(l-methyltriazol-4-yl)piperidin-4-one S65 (140 mg, 0.6081 mmol) in DCM (3 mL) was added 2-[5-(trifhioromethyl)-3-thienyl]ethanol S3 (160 mg, 0.8155 mmol), followed by MsOH (200 pL, 3.082 mmol). The mixture was refluxed for 48 hours. The mixture was cooled to room température and quenched with saturated sodium bicarbonate. The organic layer was separated and concentrated. Purification by reversed-phase HPLC (Method: Cl8 Waters Sunfire column (30x150 mm, 5 micron). Gradient: MeCN in H₂O with 0.1% trifluoroacetic acid) provided the product that was neutralized by dilution with DCM followed by saturated sodium bicarbonate wash. The solution was passed over a phase separator, and the organic layer dried to yield 2'-(difluoromethyl)-6'-(l-methyltriazol-4-yl)-2(trifluoromethyl)spiro[4,5-dihydrothieno[2,3-c]pyran-7,4'-piperidine] 389 (12 mg, 5%) as a clear oil. *H NMR (400 MHz, Chloroform-d) δ 7.49 (s, 1H), 7.16 (q, J = 1.1 Hz, 1H), 5.68 (td, J = 387

56.4, 5.1 Hz, 1H), 4.51 (dd, J =11.7, 2.7 Hz, 1H), 4.10 (s, 3H), 4.02 (t, J = 5.5 Hz, 2H), 3.64 3.54 (m, 1H), 2.75 (td, J = 5.4, 2.8 Hz, 2H), 2.43 (dt, J = 13.7, 2.7 Hz, 1H), 2.25 (dd, J = 13.4, 2.7 Hz, 1H), 1.91 (dd, J =13.6, 11.7 Hz, 1H), 1.72- 1.66 (m, 1H). LCMS m/z 409.21 [M+l]⁺

Compound 390 (2 'S,4S, 6'S, 7R)-2 '-methyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[6,7dihydrothieno[3,2-c]pyran-4,4'-piperidine]-7-ol (390)

390 C152

Step 1. Synthesis of2,2,2-trifluoro-l-[(2'S,4S,6'S)-2'-methyl-6'-(l-methyltriazol-4-yl)-210 (trifluoromethyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]-l ’-yl]ethanone (C150) To a solution of (2'S,4S,6'S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2(trifluoromethyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine] 8 (900 mg, 2.365 mmol) (Prepared via S26 intermediate using “Method B”) and DIPEA (550 pL, 3.158 mmol in DCM

388 (15 mL) at 0 °C was added TF AA (400 pL, 2.878 mmol) dropwise. After thirty minutes, the reaction was diluted with saturated sodium bicarbonate solution (10 mL), and the mixture passed over a phase separator, extracting with DCM (2x10 mL). The organics were concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-50% EtOAc in Heptane) yielded 2,2,2-trifluoro-l-[(2'S,4S,6'S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2-(trifluoromethyl)spiro[6,7dihydrothieno[3,2-c]pyran-4,4'-piperidine]-l'-yl]ethanone C150 (1.03 g, 91%) as a white foam. Ή NMR (300 MHz, Chloroformé δ 7.59 (s, 1H), 7.39 (s, 1H), 5.59 (s, 1H), 4.42 (q, J = 7.0 Hz, 1H), 4.11 (s, 3H), 3.92 (t, J = 5.5 Hz, 2H), 3.24 (dd, J = 14.8, 6.5 Hz, 1H), 2.96 - 2.73 (m, 2H), 2.48 (dd, J = 14.9, 8.4 Hz, 1H), 2.40 - 1.93 (m, 2H), 1.52 - 0.94 (m, 3H). LCMS m/z 469.07 [M+H]⁺.

Step 2. Synthesis of (2S,4S,6S)-2-methyl-6-(l-methyltriazol-4-yl)-l-(2,2,2-triftuoroacetyl)-2'(trifluoromethyl)spiro[piperidine-4,4'-thieno[3,2-cJpyranJ-7’-one (C151)

To a mixture of 2,2,2-trifluoro-l-[(2'S,4S,6'S)-2'-methyl-6'-(l-methyltriazol-4-yl)-2(trifluoromethyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]-r-yl]ethanone C150 (1.03 g, 2.153 mmol) in acetonitrile (18 mL) was added N-hydroxyphthalimide (260 mg, 1.594 mmol) and cobaltous diacetate tetrahydrate (120 mg, 0.4818 mmol), and then the mixture was vacuum purged with an oxygen balloon three times. The reaction was heated to 45 °C and stirred under an oxygen balloon atmosphère ovemight. The mixture was cooled to room température, vacuum purged with nitrogen three times, and then diluted with water (10 mL) and saturated aqueous bicarbonate (20 mL). The mixture was extracted with DCM (3 x 20 mL), and the organics were dried over sodium sulfate and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-40% EtOAc in Heptane) afforded (2S,4S,6S)-2-methyl-6-(l-methyltriazol-4-yl)-l(2,2,2-trifluoroacetyl)-2'-(trifluoromethyl)spiro[piperidine-4,4'-thieno[3,2-c]pyran]-7'-one C151 (440 mg, 42%) as a white foam. *H NMR (300 MHz, Chloroformé δ 8.03 - 7.38 (m, 2H), 5.64 (s, 1H), 4.51 - 4.30 (m, 3H), 4.13 (s, 3H), 3.48 - 3.35 (m, 1H), 2.67 (dd, J = 15.2, 8.5 Hz, 1H), 2.24 (s, 2H), 1.47 - 0.91 (m, 3H). LCMS m/z 483.11 [M+H]⁺.

Step 3. Synthesis of 2,2,2-trifluoro-l-[(2'S,4S,6'S,7R)-7-hydroxy-2'-methyl-6'-(lmethyltriazol-4-yl)-2-(trifluoromethyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4’-piperidine]l'-yl]ethenone (C152)

To apre-mixed solution of N-[(lR,2R)-2-amino-l,2-diphenyl-ethyl]-4-methylbenzenesulfonamide (1.5 mg, 0.004093 mmol) and 1,2,3,4,5pentamethylcyclopentane;rhodium(2+) tetrachloride (1 mg, 0.001592 mmol) in ACN (200 pL) was added a solution of formic acid (40 pL, 1.060 mmol) and TEA (50 pL, 0.3587 mmol). After 10 minutes, the mixture was cooled to 0 °C and a solution of (2S,4S,6S)-2-methyl-6-(l

389 methyltriazol-4-yl)-l-(2,2,2-trifluoroacetyl)-2'-(trifluoromethyl)spiro[piperidine-4,4'-thieno[3,2c]pyran]-7'-one C151 (80 mg, 0.1653 mmol) in ACN (1.5 mL) was added. The reaction was stirred at 0°C, and after one hour, it was quenched with saturated aqueous bicarbonate and extracted with DCM (2x3 mL) through a phase separator. The organics were concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-30% EtOAc in DCM) afforded 2,2,2-trifluoro-l-[(2'S,4S,6'S,7R)-7-hydroxy-2'-methyl-6'-(l-methyltriazol-4-yl)-2(trifluoromethyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]-1 '-yl]ethanone C152 (65 mg, 81%) as a colorless film. ’H NMR (300 MHz, Chloroform-7) δ 7.77 - 7.26 (m, 2H), 5.57 (s, 1H), 4.78 - 4.67 (m, 1H), 4.48 (d, J = 7.9 Hz, 1H), 4.11 (s, 3H), 3.94 (ddd, J = 41.9, 12.2, 3.7 Hz, 2H), 3.16 (dd, J = 15.0, 6.1 Hz, 1H), 2.52 (dd, J = 15.1, 8.5 Hz, 1H), 2.44 - 1.98 (m, 3H), 1.55 0.83 (m, 3H). LCMS m/z 485.09 [M+H]⁺.

Step 4. Synthesis of (2'S,4S,6'S,7R)-2'-methyl-6'-(l-methyltriazol-4-yl)-2(trifluoromethyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]-7-ol (390) To a solution of2,2,2-trifluoro-l-[(2'S,4S,6'S,7R)-7-hydroxy-2^,-methyl-6'-(lmethyltriazol-4-yl)-2-(trifluoromethyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]-ryl]ethenone C152 (65 mg, 0.1331 mmol) in MeOH (1.3 mL) was added NaOH (900 pL of 2 M, 1.800 mmol) and the mixture was heated to 50 °C. After 50 minutes, the mixture was cooled to room température, diluted with water (2 mL), and extracted with DCM (2x3 mL) through a phase separator. The organics were concentrated in vacuo to afford a clear film. The residue was brought up in DCM and heptane and stripped down to afford (2'S,4S,6'S,7R)-2'-methyl-6'-(lmethyltriazol-4-yl)-2-(trifluoromethyl)spiro[6,7-dihydrothieno[3,2-c]pyran-4,4'-piperidine]-7-ol 390 (48.3 mg, 91%) as a white foam. ’H NMR (300 MHz, Chloroform-7) δ 7.45 (s, 1H), 7.15 (s, 1H), 4.69 (s, 1H), 4.40 (dt, J = 11.8, 1.9 Hz, 1H), 4.10 - 3.93 (m, 6H), 3.45 - 3.33 (m, 1H), 2.25 (d, J = 13.9 Hz, 1H), 1.91 (d, J = 13.6 Hz, 1H), 1.81 - 1.70 (m, 1H), 1.58 (dd, J = 13.3, 11.5 Hz, 1H), 1.14 (d, J = 6.3 Hz, 3H). LCMS m/z 389.1 [M+H]⁺.

390

Compound 391

502b

DDH AIBN Chlorobenzene (Cp*RhCI₂)₂TsDPen CH₃CN Et₃N HCO₂H DCM

Step 1. To a mixture of compound 500a (8.84 g, 44.4 mmol) and 489a (9.50 g, 57.7 mmol) in DCM (250 mL) was added MsOH (34.6 mL, 532.8 mmol). The mixture was heated to 5 40 °C and stirred ovemight. TLC showed reaction completed. The mixture was cooled to 0 °C, diluted with water (200 mL) and quenched with aqueous NaOH (6 N, 100 mL). The mixture was separated, and the aqueous layer was extracted with DCM (3 x 300 mL). The combined organic layers were washed with brine (300 mL), dried over sodium sulfate, and concentrated via rotovap to afford a brown oil (crade, 20.0 g), which was further purified by flash column chromatography (DCM/acetone (1:2), then 5% MeOH/DCM and 0.5% NH₄OH) to give pure compound 501b (14.50 g, 94% yield). HPLC: 99.5% at 254 nm. LCMS: 346.20 (M+l). Ή NMR (500 MHz, CDCI3) δ 7.42 (s, 1H), 6.58 (s, 1H), 4.43 - 4.39 (m, 1H), 4.09 (m, 1H), 3.813.79 (m, 1H), 3.33-3.28 (m, 1H), 2.76-2.70 (m, 1H), 2.50-2.46 (m, 0.5H) 2.22-2.19 (m, 0.5H), 2.07-2.03 (m, 1H), 1.93-1.89 (m, 1H), 1.84 (br s, 1H), 1.68-1.63 (m, 1H), 1.44-1.40 (m, 1H),

391

1.11 (d, J= 6.5 Hz, 3H).

Step 2. To a mixture of compound 501b (11.4 g, 33.0 mmol) and DIPEA (17.2 mL, 99.0 mmol) in DCM (170 mL) in an ice-water bath was added TF AA (7.80 mL, 56.1 mmol) dropwise. After addition completed, the mixture was warmed to room température and stirred for 2 hours. TLC showed reaction completed. The reaction mixture was quenched with water (100 mL) in an ice-water bath. The biphasic mixture was stirred for 15 minutes, separated, and then the aqueous layer was extracted with additional DCM (50 mL). The combined organic layers were washed with 0.5 M aqueous HCl (50 mL), saturated NaHCO₃ (50 mL), dried over sodium sulfate, filtered, and concentrated to give a brown solid (crude 16.3 g). The solid was treated with MTBE (40 mL) and stirred at 0 °C to break up chunks for 1 hour. The white solid was collected via vacuum filtration, washed with cold MTBE (2x10 mL) and dried under high vacuum to afford compound 502b (12.6 g, 86% yield) as a white solid. HPLC: 99.0% at 230 nm. LCMS: 442.20 (M+l). 'H NMR (500 MHz, CDC1₃) δ 7.59 (s, 1H), 6.58 (s, 1H), 5.57 (m, 1H), 4.41 (m, 1H), 3.88 (dt, J= 144 Hz and 5.5 Hz, 2H), 3.17 (d, J= 144 Hz, 2H), 2.59 (d, J= 125 Hz, 2H), 2.47 (m, 1H), 2.02 (m, 1H), 1.35 (m, 3H).

Step 3. In a 250 mL three-neck round-bottom flask, compound 502b (10.70 g, 24.21 mmol) was dissolved in chlorobenzene (160 mL). The solution was sparged with nitrogen via a gas dispersion tube for 10 minutes at room température. DDH (4.85 g, 16.95 mmol) and AIBN (0.32 g, 1.94 mmol) were added. The resulting suspension was sparged with N2 for an additional 5 minutes, then heated to 75-80 °C for 4 hours. HPLC showed residual starting material (~2%), which was minor compared to the desired bromide isomers. The mixture was cooled to 20 °C, then treated with saturated NaHCCh (160 mL) and stirred for 25 minutes. The layers were separated. The aqueous layer was extracted with DCM (2 x 150 mL). The combined organic extracts were dried over sodium sulfate and concentrated via rotovap to afford a brown oil 503b (crude, 14.5 g), which was used directly in the next step.

Step 4. Crude compound 503b (14.5 g) was dissolved in DMSO (120 mL), then treated with triethylamine (15.2 mL, 109 mmol). The solution was stirred and heated to 75 °C. After 2 hours, TLC showed conversion was completed. The mixture was cooled to 20 °C, then partitioned between water (200 mL) and EtOAc (200 mL). The layers were separated. The aqueous layer was extracted with EtOAc (2 x 200 mL). The combined organic extracts were washed successively with 0.5 N aqueous HCl (150 mL), brine (150 mL), dried over sodium sulfate, filtered, and concentrated to give 11.0 g of crude solid. The solid was treated with EtOAc/MTBE (20 mL/20 mL) and stirred at room température for 1 hour to break up the chunks. The light-yellow solid was collected via vacuum filtration, washed with cold MTBE (2x10 mL) 392 and dried under high vacuum to afford compound 504b (6.40 g, 58% yield). HP LC: 95.4% at 230 nm. 'HNMR (500 MHz, DMSO-d0: δ 8.14 (s, 1H), 7.36 (s, 1H), 5.54 (m, 1H), 4.61 (m, 1H), 4.26 (m, 2H), 3.04 (m, 1H), 2.88 (m, 1H), 2.66 (m, 1H), 1.95 (m, 1H), 1.02 (m, 3H).

Step 5. To a 250 mL round-bottom flask was added (Cp*RhCb)2 (130 mg, 0.21 mmol) and TsDPEN (116 mg, 0.32 mmol) in CH3CN (50 mL). The mixture was degassed with N2 for 5 minutes. Then Et3N ( 4.2 mL, 30 mmol) and HCO2H (2.8 mL,74.8 mmol) were added. The resulting mixture was cooled in an ice-saltwater bath. To another flask, compound 504b (3.10g, 6.80 mmol) was dissolved in dry CH3CN/dry DCM (20 mL/20 mL), and the solution was degassed with N2 for 5 minutes. The resulting solution was added dropwise via dropping funnel to the above catalyst solution, keeping the internai température around 0 °C during the addition of compound 504b. After addition of compound 504b, the reaction mixture was stirred at 0 °C to 2 °C (internai temp) for 4 hours. TLC showed the reaction was almost completed. A saturated NaHCCh solution (50 mL) was added, and the cooling bath was removed. The mixture was stirred for 2 hours and extracted with EtOAc (3 x 50 mL). The combined organic extracts were washed with brine (100 mL), dried over Na2SO₄, filtered, and concentrated to dryness. The product was purified by flash column chromatography on silica gel eluting with EtOAc/ DCM (1/10 to 1/3). The product fractions were combined and concentrated in vacuo to give the product, 505b, as a white foam (2.90 g, 93% yield). HPLC: 98.3%. LCMS: 458.21 (M+l). 'H NMR (400 MHz, CDCI3): δ 7.59 (s, 1H), 6.83 (s, 1H), 5.53 (br s, 1H), 4.64 (d, J= 8.4 Hz, 0.5H), 4.47 (m, 1H), 4.25 (d, J= 8.0 Hz 0.5H), 4.16-4.00 (m, 2H), 3.80 (m, 0.5H), 3.65 (m, 0.5H), 3.26 (m, 0.5H), 2.93 (m, 0.5H), 2.81 (m, 0.5H), 2.51-2.45 (m, 1H), 2.16-2.03 (m, 1.5H), 1.29 (br s, 3H).

Step 6. To a 250 mL flask was charged compound 505b (6.44 g, 14.07 mmol) dissolved in MeOH (100 mL) and water (25 mL). Solid NaOH (6.5 g, 162.5 mmol) was added, and the stirred reaction mixture was warmed to 60 °C (oil bath température) for 3 hours. TLC showed the reaction went to completion. The reaction mixture was cooled to room température and then concentrated to dryness. The residue was purified by flash column chromatography on silica gel with MeOH/DCM/NH₄OH (7/93/0.5) to give Compound 391 (4.35 g, 86% yield) as off-white solid. HPLC: 99.4%.LCMS: 362.21 (M+l). ‘H NMR (400 MHz, DMSO-d₆): δ 7.88 (s, 1H), 6.95 (s, 1H), 5.37 (t, J= 3.2 Hz, 1H), 4.57 (d, J= 5.2 Hz, 0.5H), 4.20 (d, 7 = 5.2 Hz, 0.5H), 4.124.05 (m, 2H), 3.82-3.71 (m, 1H), 3.47-3.41 (m, 0.5H), 3.15 (d,7=4.8 Hz, 1H), 3.11-3.06 (m, 1H), 2.33-1.97 (m, 1.5H), 1.75-1.68 (m, 0.5H), 1.43-1.36 (m, 0.5H), 1.27-1.21 (m, 1H), 1.01 (d, 7= 6.4 Hz, 3H).

393

Solid State NMR Experimental (Compounds 174 and 181 Forms)

A Bruker-Biospin 400 MHz wide-bore spectrometer equipped with Bruker-Biospin 4mm HFX probe was used. Samples were packed into 4-mm ZrCh rotors and spun under Magic Angle Spinning (MAS) condition with spinning speed typically set to 12.5 kHz. The proton relaxation time was measured using *H MAS Ti saturation recovery relaxation experiment in order to set up proper recycle delay of the ¹³C and ³¹P cross-polarization (CP) MAS experiments. The fluorine relaxation time was measured using ¹⁹F MAS Ti saturation recovery relaxation experiment in order to set up proper recycle delay of the ¹⁹F MAS experiment. The CP contact time of carbon as well as phosphorus CPMAS experiments was set to 2 ms. A CP proton puise with linear ramp (from 50% to 100%) was employed. The carbon Hartmann-Hahn match was optimized on extemal reference sample (glycine), while phosphorus Hartmann-Hahn match was optimized on the actual samples. Ail carbon, phosphorus, and fluorine spectra were recorded with proton decoupling using TPPM15 decoupling sequence with the field strength of approximately 100 kHz.

Préparation of K2

K1

K2-Mg Sait

THF (3720 mL, 6.2 vol) was charged to a 5 L glass flask, then Kl (600 g, 3.47 mol, 576.92 mL, 92.6% purity by Q-NMR, 1 equiv) was added at 20°C. The mixture was cooled to 0°C and Mg(OEt)2 (198.46 g, 1.73 mol, 0.5 equiv) was charged to the reactor. The resulting mixture was stirred at 0-5°C for 10 minutes, then warmed to 20°C and stirred for 18 hours to give a milky white suspension. The hazy solution was distilled at 40°C under reduced pressure to remove THF (3.1 L). n-Hexane (3.1 L) was added, and the mixture was stirred for 2 hours to give a thick slurry. The slurry was filtered, and the filter cake was washed with n-hexane (1 x 300 mL). The solid was dried under vacuum at 40 °C for 16 hours to provide 533.6 g of K2-Mg sait (89.8% yield).

394

Préparation of K7

Step 1. K3 (600 g, 2.85 mol, 1 equiv, 96.5% purity by Q-NMR) was dissolved in anhydrous THF (3660 mL) in a 5000 mL glass flask. CDI (508.15 g, 3.13 mol, 1.1 equiv) was charged to the flask in 5 portions over 15 minutes to give a solution. The resulting reaction mixture was stirred at 18 °C for 2.5 hours. K2-Mg sait (755.77 g, 2.02 mol, 91.7% purity, 0.71 eq) was charged to the reactor in 5 portions over 8 minutes. The resulting suspension was stirred for 18 hours at 18 °C. The reaction mixture was diluted with methyl tert-butyl ether (1.8 L, 3 vol) and treated with 2 N HCl (7.1 L) to adjust the pH to 2.0 - 3.0. The organic layer was separated. The organic layer was combined and washed with saturated sodium bicarbonate (3.3 L). The organic layer was dried over anhydrous sodium sulfate and filtered, and the filtrate was evaporated at 40 °C under reduced pressure to give 862.3 g of K4 (96.9% yield).

Steps 2 and 3. A solution of K4 (570.0 g, 1.83mol, 96.7% purity by Q-NMR, 1 equiv) in dichloromethane (2850 mL, 5 vol) was cooled to 5 °C. At 0-5 °C, trifluoroacetic acid (859.15 g, 7.54 mol, 557.89 mL, 4.12 equiv = 4 eq / 0.97) was added over 80 minutes. The resulting solution was stirred at 5 °C for 1 hour, then warmed to 20 °C and stirred for 18 hours. K5 (180.76 g, 1.59 mol, 97.8% purity, 0.87 equiv) was charged as a solid in one portion, and the resulting solution was stirred for 18 hours at 20 °C. The reaction mixture was diluted with saturated brine (1.14 L, 2 vol), cooled to 5 to 10 °C, and then adjusted to pH 10 with 6 N sodium hydroxide (950 mL). The organic layer was separated and dried over sodium sulfate (400 g). The resulting solution was distilled at 30 °C under reduced pressure to remove DCM (1 L). MTBE (1.14 L) was charged, and the mixture was evaporated to dryness under reduced pressure to give an off-white solid 533.5 g. The residue was diluted with methyl tert-butyl ether (3.2 L, 6 vol) and stirred at 10-20 °C for 24 hours. The mixture was filtered, and the filter cake was washed with

395 fresh methyl tert-butyl ether (453 mL, 0.85 vol) and dried under vacuum at 45 °C for 1 hour to provide 290.4 g of K6 (62.0% yield).

Step 4. To an aqueous solution of HCl (6 M, 1.52 L, 8.83 eq) was added K6 (303 g, 1.03 mol, 1 eq) in nine portions at 30-35 °C in a 3000 mL three-necked, round-bottomed flask. The mixture was stirred at 35 °C for 1 hour. A light yellow solution was obtained. TLC and LCMS analysis indicated K6 was reacted completely. HPLC (Extemal standard method) indicated about 0.03% of K6 remained. When the reaction was completed, the mixture was cooled to 5 °C and charged with 3 g of solid K3PO4. 1.11 g of 45% KOH solution was charged portion-wise at a rate to keep the température less than 30 °C. 156 g of 45% KOH was charged resulting in a pH of 11-12. The mixture was extracted with DCM (6 x 900 mL). The combined organic layers were dried over sodium sulfate (300 g) and concentrated at 25 °C under vacuum until a heavy slurry of product was obtained. n-Heptane (200 mL) was added, and the mixture was further concentrated at 25 °C to remove solvents (200 mL). This process was repeated three times. The resulting solution was filtered, and the fïlter cake was washed with n-heptane (200 mL). The solid was dried under vacuum for 10 hours at 40 °C to give 186 g of K7 (92.9% yield).

Préparation of K8

J4 J5 J6/K8

J7

LiOH,

H₂O, THF,

Step 1 (J2): J1 (85.0 kg, 663.1 mol, 1.0 equiv) was dissolved in DMF (162.3 kg) in a

396

1000 L reactor with stirring under nitrogen and then cooled to -10 - 0 °C. NBS (122.7 kg, 689.6 mol, 1.04 eq) was dissolved in DMF (241.0 kg) in a separate, 500 L reactor with stirring under nitrogen. The solution of NBS was added slowly to the 1000 L reactor over 5 hours while maintaining the température between -10-0 °C. After the addition, the reaction mixture was held at -10-0°C for 1-2 hours. A saturated aqueous solution of NaCl (480 kg) was added to the reaction mixture followed by EtOAc (460.7 kg), and the reaction mixture was stirred for 30 minutes. The organic layer was separated and the aqueous layer was extracted with EtOAc (230.4-kg). The organic layers were combined and washed with 0.5 N HCl (420.0 kg). After séparation, a saturated solution of NaCl (300 kg) was added, and the mixture was stirred for 30 minutes. The phases were separated and the organic layer was concentrated at 40-50 °C to afford J2 (147.95 kg, 92.3% purity, 75% QNMR, 80.78% yield) as abrown liquid.

Step 2 (J3): J2 (147.95 kg, QNMR 75%, 535.8 mol, 1.0 equiv) was treated with AcOH (349.65 kg) and AC2O (82.05 kg, 803.7mol, 1.5eq) in a 1000 L reactor with stirring under nitrogen. The mixture was heated to 90-100 °C for 5-10 hours and until less than 0.5% J2 remained by GC. The mixture was cooled to 35-40 °C. NIS (138.6 kg, 616.2 mol, 1.15 eq) was added to the 1000 L reactor, and the mixture was stirred at 35-40 °C for 6-10 hours. When less than 0.5% of the intermediate remained, the mixture was cooled to 20-30 °C and transferred to a 3000 L reactor. A mixture of MTBE/heptane (250 kg/226.4 kg) and water (333 kg) were added. The mixture was stirred for 30 minutes and then separated. The aqueous layer was extracted with a mixture of MTBE/heptane (250 kg/226.4 kg). The organic layers were combined, and a 13% solution of aqueous NaHSCh (510.6 kg) was added. After stirring the mixture for 30 minutes, the layers were separated, and the organic layer was washed with 1 M NaOH (461.8kg) and water (333kg). The organic layer was concentrated at 40-60 °C to afford J3 (220.75 kg, 92.3% purity, 85.57% QNMR, 94% yield) as a brown liquid.

Step 3 (J4): J3 (111 kg, 85.57% QNMR, 252.2 mol, 1.0 equiv), Cul (12.06 kg, 63.3 mol, 0.25 equiv ), and 2,6-lutidine (6.78 kg, 63.3 mol, 0.25 equiv) were dissolved in DMAc (356.25 kg) in a 3000 L reactor with stirring under nitrogen and then heated to 85-100°C. Methyl fhiorosulfonyldifluoroacetate (MFSDA, 194.65 kg, 1013.2 mol, 4.0 equiv) was added to the 3000 L reactor while maintaining the température between 85-100°C. After the reaction mixture was held at 90-95°C for 1-4 hours, less than 5.0% J3 remained and the reaction mixture was cooled to 5-15°C. In another 3000 L reactor, water (1140 kg) and n-heptane (439.3 kg) were charged, and the mixture was cooled to 10-20 °C. The reaction was quenched to this reactor at 10-20°C, and the resulting mixture was stirred for 30 minutes. The layers were filtered and then separated. The aqueous phase was extracted with n-heptane (220 kg), and the combined organics 397 were washed with 20% NaCl (570 kg) and dried with MgSO₄ (9.5 kg, 10% w/w). The mixture was fïltered and concentrated at 35-45 °C to give crude J4. This same procedure was repeated on three additional batches of J4 (109.8 kg, QNMR 85.57%) + (110.2 kg, QNMR 85.1%) + (108.15 kg, QNMR 85.1%). The four total batches of crude J4 were combined and distilled to afford J4 (246.5 kg, 89.6 % purity, 87% QNMR, 67.7% yield) as a yellow liquid.

Step 4 (J5): NaOH (61.63 kg, 1540.8 mol, 2.28 equiv) was dissolved in water (493 kg) in a 3000 L reactor with stirring. J4 (246.5 kg, 87% QNMR, 676.2 mol, 1.0 equiv) and tetrabutylammonium bromide (TBAB, 12.33 kg, 38.25 mol, 0.057 eq) were charged, followed by 2-MeTHF (1059.95 kg). The reaction mixture was heated to 65-75°C and held at that température for 1-4 hours, at which time less than 1.0% J4 remained by HPLC analysis. The reaction mixture was cooled to 30 °C, and the phases were separated. The organic layer was washed twice with water (739.5 kg) and dried over MgSO₄ (36.98 kg). The mixture was fïltered and concentrated to dryness at 40-50 °C. n-Heptane (167.6 kg) was added, and the mixture was again concentrated to remove residual water. This process was repeated one time to afford J5 (203.2 kg, 89.46 % QNMR, 94.57% plurity,97.72% yield) as a yellow liquid.

Step 5 (J6/K8): J5 (203.2 kg, 89.46% QNMR, 660.8 mol, 1.0 equiv) was dissolved in THF (817.2 kg) in a 2000 L reactor with stirring under nitrogen. The solution was cooled to -50 to -30 °C and charged with n-BuLi (377.5 kg, 1387.7 mol, 2.1 equiv) while maintaining the température between -50 to -30 °C. After the reaction mixture was held at -50 to -30 °C for 1-2 hours, less than 1.0% J5 remained. The mixture was quenched into 20% aqueous NH₄C1 (671.9 kg) at 15 °C, and the resulting mixture was stirred for 30 minutes and separated. The aqueous phase was extracted with EtOAc (817 kg). The combined organic phases were washed twice with 20% aqueous NH₄C1 (671.9 kg), followed by 20% aqueous NaCl (408.6 kg) and then concentrated to dryness at 40-55 °C. THF (100 kg) was added, and the mixture was concentrated to remove residual water. This process was repeated one time to afford J6/K8 (147.8 kg, 89.71% purity, 83.62% QNMR, 95.41% yield) as a yellow liquid.

Step 6 (J7): J6/K8 (147.8 kg, 83.62% QNMR, 627.0 mol, 1.0 equiv) and triethylamine (95.2kg, 940.5mol, 1.5 equiv) were dissolved in THF (587.0 kg) in a 3000 L reactor with stirring under nitrogen. The mixture was cooled to -10-0 °C. 3,5-dinitrobenzoyl chloride (173.5 kg, 752.4 mol, 1.2 equiv) was dissolved in THF (587.0 kg) in a separate 1000 L reactor, and the resulting solution was transferred into the 3000 L reactor at -10-5°C. After the reaction mixture was warmed to 10-20 °C and stirred for 1.5-2 hours, less than 1.0% J6/K8 remained. 8% aqueous NaHCCh (667.4 kg) and EtOAc (500 kg) were added to the 3000 L reactor. The mixture was stirred for 30 minutes and then separated. The organic layer was washed with an 8%

398 aqueous solution of NaHCCh (667.4 kg), followed by 10% aqueous NaCl (680 kg), and then concentrated at 40-55 °C. n-Heptane (168 kg) was added, and the mixture was concentrated at 40-55 °C. EtOAc (300 kg) and n-heptane (420 kg) were added, and the mixture was heated to 65-75 °C with stirring for 1-2 hours. The slurry was cooled to 15-25°C, was stirred for 1-2 hours, and then was filtered. The solid was treated with a combination of EtOAc (450 kg) and EtOH (352 kg), and the resulting mixture was heated to 65-75°C with stirring for 1-2 hours. The mixture was cooled to 5-10°C, stirred for 1-2 hours, and filtered. The filter cake was washed with EtOH (50 kg) and dried at 40-50 °C to afford J7 (206.4 kg, 99.04% purity, 83.59% yield) as a light yellow solid.

Step 7 (K8): LiOH-HzO (66.57 kg, 1586.5 mol, 3.0 equiv) was dissolved in water (619.2 kg) in a 3000 L reactor with stirring. J7 (206.4 kg, 528.8 mol, 1.0 equiv) and THF (928.8 kg) were charged. After stirring the mixture at 30-40°C for 3 hours, less than 1% J7 remained. The layers were separated, and the THF layer was concentrated at 40-55°C. MTBE (1548 kg) was added, and the resulting mixture was washed twice with 8% aqueous NaHCO3 (668.7 kg) and then washed with 20% aqueous NaCl (743 kg). The mixture was dried over MgSO₄ (20.64kg, 10%w/w) for 1-2 hours and filtered. The organic phase was concentrated at 40-50 °C. n-Heptane (138 kg) was added, and the mixture was concentrated to remove residual MTBE. This process was repeated one time, and the resulting solution was concentrated to yield K8 (89.9 kg, 98.61% QNMR, 99.24% purity, 86.72% yield) as a light yellow, brown liquid.

Compound 181 Phosphate Sait Hydrate

MsOH, DCM, 39 °C cryst. MTBE/n-hept

TFAA, Et₃N,

DCM cryst MTBE, n-hept

399

OH ^F3^C“^lAn (RhCI₂Cp*)₂S (R.R)-TsDPEN, | I NaOH, MeOH

-------ΑμΆ-Ν. *HCO₂H, Et₃N,L V

ACN,-15°C ^F>Ao cryst: MTBE cryst: MTBE/n-hept ^F p'

K13

OH

Compound 181.H₂O Compound 181.H₃PO₄

Step 1. A solution of K7 (70 g, 0.360 mol, 1.0 equiv) and 2-[5-trifluoromethyl)-3thienyl]éthanol K8 (74.2 g, 0.378 mol, 1.05 equiv) in dichloromethane (210 mL, 3 vol) was cooled to 5 °C. Methanesulfonic acid (210.6 mL, 3.24 mol, 9 equiv) was charged to the reactor while maintaining an internai température of less than 30 °C. The resulting reaction mixture was heated to 39 °C. After 18 hours, HPLC analysis indicated greater than 99% conversion to K9. The reaction mixture was cooled to 30 °C, charged with dichloromethane (280 mL, 4 vol), and further cooled to 0 °C. The pH was adjusted to pH 10 with 4 N sodium hydroxide (830 mL). The organic layer was separated, and the aqueous phase was back-extracted with DCM (350 mL, 5 vol). The combined organics were washed with water (350 mL, 5 vol) and concentrated at reduced pressure to 3.5 total volumes. The batch was charged with MTBE (5 vol) and concentrated under reduced pressure to 3.5 total volumes. This put/take cycle was repeated three additional times, and the resulting 3.5 vol. mixture was diluted with MTBE (6.5 vol) to provide a 10 vol. mixture. The slurry was heated to 50 °C and stirred for 5 hours, then charged with nheptane (700 mL, 10 vol) over 2 hours. The resulting suspension was cooled to 20 °C over 5 hours and stirred for 18 hours. The suspension was filtered, washed with 1:2 MTBE/n-heptane (2 x 140 mL, 2x2 vol), and dried under vacuum while flushing with nitrogen at 50 °C for 18 hours to give 103 g of K9 (77% yield).

Step 2. A solution of K9 (50 g, 0.134 mol, 1.0 equiv) and triethylamine (22.5 mL, 0.161 mol, 1.2 equiv) in dichloromethane (380 mL, 7.6 vol) was cooled to 5 °C. At 5 °C, trifluoroacetic acid anhydride (20.5 mL, 0.148 mol, 1.1 equiv) was charged to the reactor while

400 keeping the internai température below 15 °C. The resulting reaction mixture was stirred at 5 °C for 1 hour, at which time HPLC showed 99.8% conversion to K10. The reaction mixture was charged at 5 °C with water (200 mL, 4 vol). The organic layer was separated and sequentially washed with 5% NaHCCh (200 mL, 4 vol), 2 N HCl (2 x 200 mL, 2x4 vol), and water (2 x 200 mL, 2x4 vol). The organic layer was concentrated under reduced pressure to 3.5 total volumes. MTBE (400 mL, 8 vol) was charged, and the batch was concentrated under reduced pressure to 3.5 vol. This put/take cycle was repeated two additional times, and the mixture was concentrated to 3 volumes after the final cycle. The solution was heated to 40 °C and charged with n-heptane (190 mL, 2 vol) over 1 hour. The batch was cooled to 20 °C over 2 hours to yield a suspension. n-Heptane (500 mL, 10 vol) was charged over 2 hours, and the resulting suspension was stirred for 18 hours. The suspension was filtered, washed with 5% MTBE/n-heptane (2 x 125 mL, 2 x 2.5 vol), and dried under vacuum while flushing with nitrogen at 50 °C for 18 hours to give 53 g of K10 (84% yield).

Step 3. A suspension of K10 (70 g, 149.4 mmol, 1.0 equiv), and l,3-dibromo-5,5’dimethylhydantoin (29.9 g, 104.6 mmol, 0.7 equiv) in anhydrous chlorobenzene (280 L, 4 vol) was sparged with sub-surface nitrogen bubble for 60 minutes. The mixture was heated to 75 °C and charged at that température with a prepared solution of azobisisobutyronitrile (0.49 g, 3 mmol, 0.02 equiv) in anhydrous chlorobenzene (70 mL, 1 vol) over 60 minutes. After stirring for 2 hours at 75 °C, HPLC analysis showed conversion to Kll. The reaction mixture was cooled to 60 °C and charged with anhydrous, degassed DMSO (350 mL, 5 vol) over 30 minutes, followed by anhydrous, degassed triethylamine (104 mL, 747 mmol, 5 equiv) over 30 minutes. The reactor headspace was well purged with nitrogen, and the batch was heated to 75 °C. After 15 hours, HPLC analysis showed >99% conversion of Kll to K12. The batch was cooled to 20 °C and diluted with dichloromethane (210 mL, 3 vol). The batch was further cooled to 5 °C and charged with water (350 mL, 5 vol) while keeping the solution température below 30 °C. The organic layer was separated, and the aqueous layer was back-extracted with dichloromethane (210 mL, 3 vol). The organic phases were combined and washed sequentially with 2 N HCl (350 mL, 5 vol) and water (2 x 350 mL, 2x5 vol). The organic phase was concentrated under reduced pressure to 3 total volumes. The solution was charged with IPA (560 mL, 8 vol) and concentrated under reduced pressure to 3 volumes. This put/take cycle was repeated two additional times, giving a 3-volume solution that was further diluted with IPA (70 mL, 1 vol). The resulting 4 vol mixture was heated to 75 °C to provide a homogenous solution and then cooled to 50 °C. The solution was seeded (0.1 wt%) at 50 °C, stirred for 1 hour, and further cooled to 20 °C over 2 hours. After stirring an additional 18 hours at 20 °C, the slurry was

401 charged with n-heptane (70 mL, 1 vol) over 1 hour. The slurry was stirred for 4 hours at 20 °C, filtered, washed with 1:1 IPA/n-heptane (2 x 70 mL, 2x2 vol), and dried under vacuum while flushing with nitrogen at 50 °C for 18 hours to give 31.2 g of K12 (43% yield from K10). The dried K12 was suspended in IPA (93 mL, 3 vol), heated to 80 °C, and stirred at that température for 2 hours. The solution was cooled to 70 °C over 1 hour and stirred for 1 hour. The suspension was cooled to 20 °C over 5 hours and stirred at that température for 18 hours. The suspension was filtered, washed with 1:1 IPA/n-heptane (2x35 mL, 2 x 0.5 vol), and dried under vacuum while flushing with nitrogen at 50 °C for 18 hours to give 28.8 g of K12 (40% yield from K10).

Step 5. The pentamethylcyclopentadienylrhodium(III)chloride dimer (154 mg, 0.002 eq) and (R,R)-TsDPEN (182 mg, 0.004 eq) were combined in acetonitrile (240 mL, 4 vol), and the mixture was sparged with nitrogen while stirring at 20 °C for 1 hour. The mixture was cooled to -15 °C and a prepared mixture of formic acid (27.0 mL, 5.5 equiv) and triethylamine (38.1 mL, 2.2 eq) was added over 30 minutes and the résultant red/orange solution was stirred for 15 minutes at -15 °C. A solution of K12 (60 g, 1.0 equiv) in acetonitrile (240 mL, 4 vol) was separately prepared and added to the cold catalyst solution over 45 minutes. The mixture was sparged with subsurface nitrogen bubble for 15 minutes, stirred at -15 °C for 20 hours, warmed to 0 °C, and stirred for an additional 20 hours. The température was adjusted to 20 °C and the mixture was charged with MTBE (360 mL, 6 vol) and 18% NaCl (aq) (360 mL, 6 vol). The phases were mixed, and the phases separated. The organic phase was washed sequentially with 18% NaCl (aq) (2 x 360 mL, 6 vol), 4% NaHCO₃ (aq) (360 mL, 6 vol), and 18% NaCl (aq.) (180 mL, 3 vol). The reaction solution was concentrated to 3 total volumes under reduced pressure, then solvent swapped to MTBE by adding MTBE (360 mL, 6 vol) and concentrating to 3 volumes under reduced pressure. This put/take cycle was repeated 3 additional times. The resulting solution was diluted to 4 total volumes with MTBE and charged with DCM (240 mL, 4 vol) and MTBE-pre-washed SiliaMetS DMT resin (30 g, 50 wt%). The mixture was stirred vigorously at 20 °C for 2 hours. The resin slurry was filtered under vacuum. The reaction flask was rinsed with a solution of 2:1 DCM:MTBE (120 mL, 2 vol) and the rinse was transferred to the resin. The resulting slurry was mixed, then filtered under vacuum. The resin was rinsed once more with a solution of 2:1 DCM:MTBE (120 mL, 2 vol) by adding it to the resin, mixing, then filtering under vacuum. The rinses and original filtrate were combined and transferred back to the reaction flask using 2:1 DCM:MTBE (30 mL, 0.5 vol) as a final rinse after the transfer. The filtrate was combined with MTBE-pre-washed SiliaMetS DMT resin (30 g, 50 wt%) and stirred vigorously for 2 hours at 20 °C. The resin slurry was under vacuum. A solution of 2:1

DCM:MTBE (120 mL, 2 vol) was used to rinse the reaction flask, and the rinse was transferred

402 to the resin in the frit. The slurry was mixed and filtered under vacuum. A solution of 2:1 DCM:MTBE (120 mL, 2 vol) was charged to the resin in the frit and the slurry was mixed, then filtered under vacuum. The combined filtrâtes were transferred back to the reaction flask using 2:1 DCM:MTBE (30 mL, 0.5 vol) as a rinse. The filtrate was combined with MTBE-pre-washed SiliaMetS DMT resin (30 g, 50 wt% loading) and stirred vigorously for 18 hours. The résultant resin slurry was filtered under vacuum. The reaction flask was rinsed with a solution of 2:1 DCM:MTBE (120 mL, 2 vol). The rinse was added to the resin in the frit, and the slurry was mixed then filtered under vacuum. A solution of 2:1 DCM:MTBE (120 mL, 2 vol) was added to the resin in the frit, and the slurry was mixed then filtered under vacuum. The combined filtrate was transferred to a flask, then concentrated to 3 total volumes (180 mL) of solution. MTBE (480 mL, 8 vol) was added, and the solution was concentrated to 3 total volumes (180 mL). This put/take cycle was repeated two additional times. The resulting solution was diluted to 5 vol (300 mL) with MTBE, heated to 50 °C and stirred for 3 hours. n-Heptane (240 mL, 4 vol) was added over 60 minutes, and the slurry was maintained at 50 °C for an additional 1 hour. The slurry was cooled to 20 °C over 3 hours and stirred ovemight. The slurry was filtered under vacuum. The cake was rinsed with 1:1 MTBE:heptane (2 x 60 mL, 2x1 vol), and the solids were dried under vacuum at 50 °C for 18 hours to yield 58.5 g of K13 (83% yield).

Step 6. K13 (43.5 g, 89 mmol, 1 equiv) and methanol (150.0 mL, 3 vol) were combined and agitated until full dissolution was observed. 6 N NaOH (89 mL, 6 eq) was added drop-wise over 30 minutes, and the mixture was heated to 60 °C and stirred for 1 hour at which time complété conversion to Compound 181 was achieved. The reaction solution was cooled to 15 °C and treated with isopropyl acetate (250 mL, 5.75 vol). Water (100 mL, 2.3 vol) was then added, and the mixture was agitated for 30 minutes. The phases were separated, and the aqueous phase was back-extracted with isopropyl acetate (250 mL, 5.75 vol). The organics were combined and washed with 10% NaCl (aq.) (2 x 250 mL, 2 x 5.75 vol) and water (250 mL, 5.75 vol). The organics were concentrated to 4.0 total volumes (174 mL). The solution was charged with MTBE (11.5 vol, 500 mL) and concentrated again to 4.0 vol. This put/take cycle was repeated three additional times. MTBE (75 mL, 1.75 vol) was added to give a 5.75 vol, 250 mL, solution. While stirring at 20 °C, water (3.2 mL, 180 mmol, 2 eq) was added over 2 hours, inducing crystallization. The slurry was stirred at 20 °C for 1 hour, then heated to 50 °C and stirred at that température for 3 hours. The suspension was cooled to 20 °C and stirred for 18 hours. The slurry was filtered under vacuum, and the cake was washed with MTBE (100 mL, 2.3 vol). The solids were dried at 50 °C under vacuum for 18 hours to provide 29 g of Compound 181 Free Form Monohydrate (Compound I8I.H2O) (81% yield).

403

Step 7. Method A. 1 eq. of Compound 181 free form monohydrate was charged to a reactor followed by 6 vol. of MEK. Agitation was started at 20 °C. Once a clear solution was obtained, the solution was polish filtered and charged back to the reactor. Water (0.2 vol.) was added to the clear solution and agitation continued. 1 wt% of Compound 181 Phosphate Sait was added as seeds. In a separate container, 1.02 eq. of 85 wt% phosphoric acid was diluted with 3.8 vol. of MEK. This phosphoric acid solution was then added to the reactor slowly over 3 hours. The final slurry was agitated at 20 °C for 2 hours, then filtered under vacuum. The resulting wet cake was washed with 3 vol. of MEK. The wet cake was dried under vacuum with a nitrogen bleed at 80°C to yield Compound 181 Phosphate Sait Hydrate (Compound I8I.H3PO4) (about 90% yield).

Method B. 1 eq. of Compound 181 free form monohydrate was charged to a reactor, followed by 6 vol. of MEK. Agitation was commenced at 20 °C, and once a clear solution was obtained, the solution was polish filtered and charged back to the reactor. Water (0.2 vol.) was added to the clear solution and agitation continued. In a separate container, 1.02 eq. of 85 wt% phosphoric acid was diluted with 3.8 vol. of MEK. This phosphoric acid solution was then added to the reactor slowly over 3 hours. The final slurry was agitated at 20 °C for 2 hours then filtered under vacuum. The resulting wet cake was washed with 3 vol. of MEK. The wet cake was dried under vacuum with a nitrogen bleed at 80°C to yield Compound 181 Phosphate Sait Hydrate (Compound I8I.H3PO4) (about 90% yield).

Note: Compound 181 Phosphate Sait Hydrate is a crystalline hydrate.

XRPD and VH-XRPD

The powder, X-ray powder diffraction (XRPD), diffractogram of Compound 181 Phosphate Sait Hydrate (FIG. 1) was acquired at room température (25 ± 2 °C) in transmission mode using a PANalytical Empyrean System equipped with a sealed tube source and a PIXcel 1D Medipix-3 detector (Malvem PANalytical Inc, Westborough, Massachusetts). The X-Ray generator operated at a voltage of 45 kV and a current of 40 mA with copper radiation (1.54060 Â). The powder was placed in sample stage AP CHC stage and CHC chamber. The CHC chamber was attached to a water pump which collected the relative humidity. The relative humidity in the chamber was stepwise changed in incréments, starting at 5% for 1 hour, then increased to 10% and held for an hour followed by 10% relative humidity (RH) stepwise increased to 60% and held for an hour at each, with a jump at 60% to 90% and held for 1 hour. The CHC Chamber was then held at 90% for an additional hour, then decreased from 90% to 80% and held for 3 hours, then from 80% to 70% and held for 3 hours, then from 70% to 60%

404 and held for 3 hours, then from 60% to 10% decreased stepwise by 10% RH and held for an hour at each step and last decreased from 10% to 5% and held for an hour. At the hour time point, XRPD collection was run over the range of about 3° to about 40° 20 with a step size of 0.0131303° and 49s per step.

Variable Humidity XRPD (VH-XRPD): Compound 181 Phosphate Sait Hydrate was observed to hâve continuous peak shift which are ail (within ± 0.2 °20) from 5-90% relative humidity (FIG. 2, Table 14).

Table 14. Peak List from XRPD Diffractogram of Compound 181 Phosphate Sait Hydrate

XRD Peaks	Relative Humidity 40%	Relative Humidity 5%	Relative Humidity 90%
Angle (°20 ±0.2)	Intensity %	Angle (°20 ±0.2)	Intensity %	Angle (°20 ±0.2)	Intensity %
1	19.9	100.0	19.9	100.0	19.9	100.0
2	8.6	76.2	8.6	79.2	8.6	65.3
3	28.3	64.3	28.3	69.4	28.3	60.9
4	20.4	56.7	20.4	61.9	20.4	55.2
5	21.0	43.0	22.8	37.1	21.0	48.7
6	22.8	41.4	17.2	31.9	27.8	44.9
7	17.2	38.3	21.9	30.1	22.8	40.9
8	27.8	37.2	21.1	29.7	17.2	40.5
9	26.4	28.4	27.0	29.3	19.5	30.9
10	17.8	27.2	15.7	23.7	25.5	30.6
11	15.7	26.8	27.8	22.9	17.8	29.2
12	25.5	26.2	25.8	18.2	15.8	26.6
13	19.5	25.8	16.9	17.6	21.9	25.8
14	21.9	25.5	17.8	17.1	16.9	24.1
15	27.1	23.3	19.6	16.7	27.1	23.2
16	16.9	22.7	26.4	15.9	26.4	22.5
17	21.7	20.1	25.1	15.2	25.1	20.0
18	25.1	19.4	25.4	15.1	25.9	16.1
19	25.9	16.6	22.1	14.3	25.3	15.1
20	19.7	14.7	17.7	14.2	13.0	13.7
21	22.0	13.6	12.9	12.6	20.6	13.4

405

XRD Peaks	Relative Humidity 40%	Relative Humidity 5%	Relative Humidity 90%
Angle (°20 ±0.2)	Intensity %	Angle (°20 ±0.2)	Intensity %	Angle (°20 ±0.2)	Intensity %
22	13.0	13.1	18.5	12.2	18.5	12.1
23	25.3	12.7	27.4	11.6	11.5	10.4
24	18.5	12.3	11.5	11.5	17.6	10.4
25	17.6	11.9			27.4	10.4
26	11.5	11.5			13.1	10.2
27	27.4	11.0
28	13.2	10.1

TGA

Thermal gravimétrie analysis of Compound 181 Phosphate Sait Hydrate was performed using the TA Instruments Q5000 TGA. A sample with a weight of approximately 1-10 mg was scanned from ambient to 250 °C at a heating rate of 10 °C/min with nitrogen purge. The TGA thermogram shows around 0.5% weight loss from ambient température up until 150 °C (FIG. 3).

DSC

Differential Scanning Calorimetry (DSC) analysis of Compound 181 Phosphate Sait Hydrate was performed using the TA Instruments Q2000 DSC. A sample with a weight between 1-10 mg was weighed into an aluminum crimp sealed pan with a pinhole. This pan was placed in the sample position in the calorimeter cell. An empty pan was placed in the reference position. The calorimeter cell was closed, and a flow of nitrogen was passed through the cell. The program was set to modulate 0.32° per 60 seconds, then heat rate at of 2 °C per min to a température to 300 °C. The thermogram shows two endotherm peaks around 226 °C and 251 °C (FIG. 4).

SSNMR

The ¹³C CPMAS of Compound 181 Phosphate Sait Hydrate (FIG. 5, Table 15) was acquired at 275K and 43% relative humidity (RH) with 12.5 kHz spinning and using adamantane as a reference.

406

Table 15. Peak List from ¹³C CPMAS of Compound 181 Phosphate Sait Hydrate

Peak #	Chem Shift [ppm]	Intensity [rel]
1	146.3	42.1
2	145.8	45.6
3	144.0	42.5
4	141.7	58.3
5	139.3	51.8
6	129.4	46.4
7	128.6	52.3
8	126.6	46.8
9	73.6	87.5
10	73.2	83.2
11	66.1	38.9
12	64.3	43.7
13	62.7	55.1
14	62.1	62.3
15	48.9	44.6
16	47.3	70.8
17	45.4	50.6
18	43.0	39.6
19	41.6	48.8
20	38.4	100.0
21	36.7	48.3
22	16.0	94.4

The ¹⁹F MAS of Compound 181 Phosphate Sait Hydrate (FIGs. 6, 7; Tables 16, 17) was acquired at 275K and 0%, 6%, 22%, 43%, 53%, 75%, and 100% relative humidity (RH) with

12.5 kHz spinning and using adamantane as a reference.

Table 16. Peak List from ¹⁹F MAS of Compound 181 Phosphate Sait Hydrate at 43% RH

Peak#	Chem Shift [ppm]	Intensity [rel]
1	-53.8	11.0

407

2

-57.4

12.5

Table 17. Effect of Relative Humidity on ¹⁹F MAS of Compound 181 Phosphate Sait Hydrate

RH [%]	Peak 1 [ppm]	Peak 2 [ppm]
0	-53.4	-57.6
6	-53.6	-57.5
22	-53.8	-57.5
33	-53.8	-57.4
43	-53.8	-57.4
53	-53.8	-57.4
75	-53.9	-57.4
100	-53.9	-57.4

The ³¹P CPMAS of Compound 181 Phosphate Sait Hydrate (FIGs. 8, 9; Tables 18, 19) was acquired at 275K and 0%, 6%, 22%, 43%, 53%, 75%, and 100% relative humidity (RH) with 12.5 kHz spinning and using adamantane as a reference.

Table 18. Peak List from ³¹P CPMAS of Compound 181 Phosphate Sait Hydrate at 43%

RH

Peak#	Chem Shift [ppm]	Intensity [rel]
1	4.2	46.4
2	2.6	100.0

Table 19. Effect of Relative Humidity on ³¹P CPMAS of Compound 181 Phosphate Sait Hydrate

RH [%]	Peak 1 [ppm]	Peak 2 [ppm]
0	6.1	2.6
6	5.1	2.6
22	4.4	2.6
33	4.2	2.6
43	4.2	2.6
53	4.1	2.5
75	4.0	2.5
100	3.8	2.5

408

Alternative Préparation of Compound 181 Free Form Monohydrate

Amorphous Compound 181 (30 mg) was added to saline (1 mL). After mild vortexing to see if the material would dissolve, a white milky precipitate formed. The sample was left ovemight at ambient température. The solid material was filtered using a 0.22 pm PVDF Eppendorf filter tube, rinsing with ice cold water. The sample was dried in a vacuum oven at 45°C ovemight. Both the wet cake and dried materials were crystalline Compound 181 Free Form Monohydrate.

XRPD

The powder, X-ray powder diffraction (XRPD), diffractogram of Compound 181 Free Form Monohydrate was acquired at room température in transmission mode using a PANalytical Empyrean System equipped with a sealed tube source and a PIXcel 1D Medipix-3 detector (Malvem PANalytical Inc, Westborough, Massachusetts) (FIG. 10, Table 20). The X-Ray generator operated at a voltage of 45 kV and a current of 40 mA with copper radiation (1.54060 Â). The powder sample was placed on a 96 well sample holder with mylar film and loaded into the instrument. The sample was scanned over the range of about 3° to about 40°20 with a step size of 0.0131303 and 49s per step.

Table 20. Peak List from XRPD Diffractogram of Compound 181 Free Form Monohydrate

XRD Peaks	Angle (°20 ±0.2)	Intensity %
1	16.7	100.0
2	21.7	37.0
3	8.7	23.3
4	12.8	18.4
5	19.8	15.9
6	25.8	15.8
7	13.8	15.1
8	15.5	12.7
9	24.3	12.7

TGA

Thermal gravimétrie analysis of Compound 181 Free Form Monohydrate was performed using the TA5500 Discovery TGA. A sample with a weight of approximately 1-10 mg was scanned from ambient température to 250 °C at a heating rate of 10 °C/min with nitrogen purge.

409

The TGA thermogram showed around ~3-4% weight loss from ambient température up until 100 °C (FIG. 11).

DSC

The Differential Scanning Calorimetric analysis of Compound 181 Free Form Monohydrate was performed using the TA Instruments Q2000 DSC. A sample with a weight between 1-10 mg was weighed into an aluminum crimp sealed pan with a pinhole. This pan was placed in the sample position in the calorimeter cell. An empty pan was placed in the reference position. The calorimeter cell was closed, and a flow of nitrogen was passed through the cell. The program was set to modulate 0.32° per 60 seconds, then heated at a rate of 2 °C per min to a température to 300 °C. The thermogram showed three endotherm peaks around 61 °C, 94 °C, and 111°C (FIG. 12).

SSNMR

The ¹³C CPMAS of Compound 181 Free Form Monohydrate (FIG. 13, Table 21) was acquired at 275K and 43% relative humidity (RH) with 12.5 kHz spinning and using adamantane as a reference. Additionally, the ¹³C CPMAS of Compound 181 Free Form Monohydrate following déhydration (80 °C in rotor ovemight (2x), 80 °C weekend incubation with P2O5) (FIG. 14, Table 22) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference.

Table 21. Peak List from ^I3C CPMAS of Compound 181 Free Form Monohydrate

Peak#	Chem Shift [ppm]	Intensity [rel]
1	149.6	57.4
2	149.4	33.3
3	135.3	63.4
4	129.6	28.0
5	127.7	23.3
6	126.2	26.9
7	74.4	100.0
8	68.1	40.7
9	61.6	47.1
10	49.8	47.2

410

Peak#	Chem Shift [ppm]	Intensity [rel]
11	47.8	33.0
12	47.0	36.0
13	39.3	41.7
14	35.1	43.2
15	24.9	55.8

Table 22. Peak List from ¹³C CPMAS of Dehydrated Compound 181 Free Form Monohydrate

Peak#	Chem Shift [ppm]	Intensity [rel]
1	150.9	33.4
2	150.0	53.9
3	135.3	64.6
4	129.6	30.1
5	127.2	29.2
6	126.6	32.6
7	74.7	100.0
8	68.4	14.9
9	61.5	38.6
10	50.7	48.5
11	48.8	22.2
12	48.3	41.7
13	47.5	23.4
14	47.2	41.7
15	36.8	45.2
16	35.8	42.5
17	25.6	56.2

The ¹⁹F MAS of Compound 181 Free Form Monohydrate (FIG. 15, Table 23) was acquired at 275K and 43% relative humidity (RH) with 12.5 kHz spinning and using adamantane as a reference, with ^I9F background subtracted. Additionally, the ¹⁹F MAS of Compound 181 Free Form Monohydrate following déhydration (80 °C in rotor ovemight (2x), 80 °C weekend

411 incubation with P2O5) (FIG. 16, Table 24) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference, with ¹⁹F background subtracted.

Table 23. Peak List from ¹⁹F MAS of Compound 181 Free Form Monohydrate

Peak #	Chem Shift [ppm]	Intensity [rel]
1	-55.8	12.5

Table 24. Peak List from ¹⁹F MAS of Dehydrated Compound 181 Free Form Monohydrate

Peak#	Chem Shift [ppm]	Intensity [rel]
1	-55.5	12.5

Compound 181 Phosphate Sait Methanol Solvaté

Amorphous Compound 181 (50 mg) was added to MEK (0.3 mL). To this was added 0.27 mL of a 0.5 M stock solution of H3PO4 in MeOH. The sample was lefit at ambient température ovemight. The solids were filtered using a 0.22 μιη PVDF Eppendorf filter tube and washed with 4:1 n-Heptane/MEK (v/v) that was chilled over ice. Subséquent washes were performed with n-Heptane, resulting in a solid white powder. XRPD of the wet material showed the product was Compound 181 Phosphate Sait Methanol Solvaté.

XRPD

The powder, X-ray powder diffraction (XRPD), diffractogram of Compound 181 Phosphate Sait Methanol Solvaté (FIG. 17, Table 25) was acquired at room température in transmission mode using a PANalytical Empyrean System equipped with a sealed tube source and a PIXcel 3D Medipix-3 detector (Malvem PANalytical Inc, Westborough, Massachusetts). The X-Ray generator operated at a voltage of 45 kV and a current of 40 mA with copper radiation (1.54060 Â). The powder sample was placed on a 96 well sample holder with mylar film and loaded into the instrument. The sample was scanned over the range of about 3° to about 40°20 with a step size of 0.0131303° and 49s per step.

Table 25. Peak List from XRPD Diffractogram of Compound 181 Phosphate Sait Methanol Solvaté

XRD Peaks	Angle (°20 ±0.2)	Intensity %
1	15.8	100.0

412

XRD Peaks	Angle (°20 ±0.2)	Intensity %
2	20.7	89.2
3	12.7	59.5
4	8.5	54.2
5	19.5	45.5
6	18.7	36.8
7	13.9	35.6
8	10.2	30.3
9	22.5	29.5
10	21.5	27.4
11	3.9	26.4
12	20.0	24.9
13	19.2	24.5
14	24.9	24.0
15	19.6	23.3
16	21.8	21.5
17	27.4	21.3
18	12.9	21.0
19	25.2	20.8
20	14.8	20.7
21	17.3	18.0
22	9.6	17.8
23	20.4	17.0
24	17.6	15.9
25	16.0	15.5
26	11.4	13.9
27	18.4	13.7
28	25.5	12.5
29	27.9	12.2
30	27.6	11.1
31	12.5	10.8
32	23.5	10.7

413

SSNMR

The ¹³C CPMAS of Compound 181 Phosphate Sait Methanol Solvaté (FIG. 18, Table 26) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference.

Table 26. Peak List from ¹³C CPMAS of Compound 181 Phosphate Sait Methanol Solvaté

Peak#	Chem Shift [ppm]	Intensity [rel]
1	146.8	54.0
2	145.8	50.8
3	143.9	47.8
4	140.6	82.3
5	139.5	66.0
6	129.4	71.6
7	128.5	56.2
8	127.9	58.2
9	126.7	46.5
10	73.8	94.9
11	72.2	95.2
12	66.3	66.8
13	64.2	61.7
14	62.8	69.1
15	61.6	77.9
16	49.7	56.9
17	48.5	80.3
18	47.1	100.0
19	45.5	57.9
20	43.0	51.0
21	40.5	73.1
22	40.1	65.6
23	38.9	66.2
24	37.7	62.1
25	36.8	58.6
26	17.7	78.3

414

Peak#	Chem Shift [ppm]	Intensity [rel]
27	15.7	78.5

The ¹⁹F MAS of Compound 181 Phosphate Sait Methanol Solvaté (FIG. 19, Table 27) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference, with ¹⁹F background subtracted.

Table 27. Peak List from ¹⁹F MAS of Compound 181 Phosphate Sait Methanol Solvaté

Peak#	Chem Shift [ppm]	Intensity [rel]
1	-54.7	11.8
2	-57.7	12.5

The ³¹P CPMAS of Compound 181 Phosphate Sait Methanol Solvaté (FIG. 20, Table 28) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference.

Table 28. Peak List from ³¹P CPMAS of Compound 181 Phosphate Sait Methanol Solvaté

Peak#	Chem Shift [ppm]	Intensity [rel]
1	2.5	93.9
2	1.8	100.0

Compound 181 Phosphate Sait MEK Solvaté

Compound 181 Phosphate Sait Hydrate (25 mg) was added to 2-butanone (MEK) (1 mL) in an HP LC vial. The sample was mixed and formed a slurry. The slurry was placed in a cold room at 5 °C with a small stir bar for 11 days. The solid material was centrifuged and fïltered using a 0.22 pm PVDF Eppendorf filter tube at room température. The XRD of the wet cake sample showed that it was Compound 181 Phosphate Sait MEK Solvaté.

XRPD

The powder, X-ray powder diffraction (XRPD), diffractogram of Compound 181 Phosphate Sait MEK Solvaté was acquired at room température (25 ± 2 °C) in transmission mode using a PANalytical Empyrean System equipped with a sealed tube source and a PIXcel 1D Medipix-3 detector (Malvem PANalytical Inc, Westborough, Massachusetts) (FIG. 21, Table 29). The X-Ray generator operated at a voltage of 45 kV and a current of 40 mA with

415 copper radiation (1.54060 Â). The powder sample was placed on a 96 well sample holder with mylar film as well Kapton tape over the sample and loaded into the instrument. The sample was scanned over the range of about 3° to about 40°20 with a step size of 0.0131303° and 49s per step.

Table 29. Peak List from XRPD Diffractogram of Compound 181 Phosphate Sait MEK Solvaté

XRD Peaks	Angle (°20 ±0.2)	Intensity %
1	20.1	100.0
2	15.4	85.7
3	8.6	80.8
4	15.7	36.5
5	19.4	32.1
6	18.2	32.0
7	21.7	30.8
8	21.9	29.0
9	13.2	28.6
10	23.8	25.9
11	10.8	25.1
12	10.5	24.1
13	21.0	23.0
14	22.8	21.7
15	17.5	18.8
16	18.4	18.2
17	26.7	16.8
18	22.4	14.4
19	3.8	12.4
20	8.3	11.0
21	16.5	10.6

SSNMR

The ¹³C CPMAS of Compound 181 Phosphate Sait MEK Solvaté (FIG. 22, Table 30) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference.

416

Table 30. Peak List from ¹³C CPMAS of Compound 181 Phosphate Sait MEK Solvaté

Peak#	Chem Shift [ppm]	Intensity [rel]
1	146.6	36.9
2	145.8	41.2
3	144.1	34.3
4	143.6	32.3
5	142.0	55.2
6	140.9	20.2
7	139.4	41.6
8	138.4	26.0
9	130.7	16.3
10	129.6	52.8
11	128.7	46.9
12	128.0	32.6
13	126.5	54.5
14	73.7	79.9
15	73.2	86.8
16	66.3	60.6
17	64.3	35.0
18	63.3	25.6
19	62.7	48.2
20	62.3	73.8
21	50.4	31.2
22	48.8	54.4
23	48.4	32.8
24	47.4	57.8
25	46.3	24.0
26	45.5	42.2
27	44.3	23.2
28	43.3	31.7
29	42.3	28.9
30	41.9	40.4

417

Peak#	Chem Shift [ppm]	Intensity [rel]
31	38.4	100.0
32	37.5	56.0
33	36.8	48.4
34	35.3	23.6
35	29.5	24.2
36	16.0	74.8
37	15.2	33.7
38	7.4	44.5

The ¹⁹F MAS of Compound 181 Phosphate Sait MEK Solvaté (FIG. 23, Table 31) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference, with ¹⁹F background subtracted.

Table 31. Peak List from ¹⁹F MAS of Compound 181 Phosphate Sait MEK Solvaté

Peak#	Chem Shift [ppm]	Intensity [rel]
1	-53.6	10.0
2	-55.2	5.2
3	-57.2	12.5

The ³¹P CPMAS of Compound 181 Phosphate Sait MEK Solvaté (FIG. 24, Table 32) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference.

Table 32. Peak List from ³¹P CPMAS of Compound 181 Phosphate Sait MEK Solvaté

Peak#	Chem Shift [ppm]	Intensity [rel]
1	4.8	94.7
2	2.7	15.7
3	0.1	100.0

Alternative Préparation of Compound 181 Phosphate Sait Hydrate

2.05 g Compound 181 Phosphate Sait Methanol Solvaté was dried at 50 °C for 21 hours with N2 purge. The résultant solid was Compound 181 Phosphate Sait Hydrate.

418

Compound 174 Hemihydrate

MsOH, DCM, 39 °C cryst MTBE/n-hept

K8

TFAA, Et₃N,

DCM cryst. MTBE, n-hept

OH

K17 (RhCI₂Cp*)₂(R.R)-TsDPEN, HCO₂H, Et₃N, ACN, -15 °C

NaOH, MeOH

K18

OH

Compound 174

OH

Compound 174.1/2 H₂O

Step 1. A solution of K7 (4153 g, 1 equiv, 81.11% purity by Q-NMR, 21.53 mmol, 1 equiv) and K8 (3651 g, 22.45 mmol, 1.05 equiv) in dichloromethane (33.2L, 8 vol) was treated with methanesulfonic acid (14384 g, 149.7 mol, 7 equiv) at 0 °C over 1 hour. The resulting

419 mixture was heated at 40 °C. After 14 hours, analysis showed >99% consumption of K7. The reaction mixture was cooled to 10 °C and adjusted to pH 10 with 4 N sodium hydroxide (40 L). The organic layer was separated, dried over sodium sulfate (1.5 kg), and evaporated under reduced pressure at 25 °C to give crude K14 as an off-white solid (8.1 kg). This solid was suspended in methyl ZerZ-butyl ether (22 L), stirred at 10 °C for 2.5 hours, and then filtered. The filter cake was washed with methyl ZerZ-butyl ether (4 L) and dried under vacuum while flushing with nitrogen at 20 °C for 18 hours to give purified 5950 g K14 (96.8% yield).

Step 2. A solution of K14 (5937 g, 17.52 mol, 1 equiv) and ΛζΑ-diisopropylethylamine (3967 mL, 22.78 mol, 1.3 equiv) in dichloromethane (59 L, 10 vol) was cooled to 0-5 °C and treated with trifluoroacetic acid anhydride (2680 mL, 19.27 mol, 1.1 equiv) over 40 minutes while keeping the reaction température below 14 °C. The resulting reaction mixture was stirred at 0-10 °C. After 2 hours, HP LC analysis indicated >99.5% conversion. The reaction mixture was cooled to 5 °C and diluted with saturated brine (27 L). The resulting mixture was adjusted to pH 10 with 6 N sodium hydroxide solution (5 L) while keeping the température below 12 °C. The resulting mixture was stirred for 20 minutes, then the layers were separated. The organic layer was sequentially washed with 2 N HCl (3 x 22 L), water (3 x 22 L), and brine (22 L), then dried over sodium sulfate (1 kg) and evaporated under reduced pressure at 30 °C to give crude K15 (7519 g). The crude material was suspended in a mixture of methyl ZerZ-butyl ether (16 L) and n-heptane (8 L) at 50 °C for 5 hours, then cooled to 20 °C over 5 hours. After 18 hours of stirring at 20 °C, the suspension was filtered. The filter cake was washed with a mixture of methyl ZerZ-butyl ether (8 L) and n-heptane (4 L), then dried under vacuum while flushing with nitrogen at 20 °C for 18 hours to give 6824 g of K15 (94.1% yield).

Steps 3 and 4. A suspension of K15 (5879 g, 13.52 mol, 1 equiv), azobisisobutyronitrile (178 g, 1.082 mol, 0.08 equiv), and l,3-dibromo-5,5-dimethylhydantoin (2900 g, 10.14 mol, 0.75 equiv) in chlorobenzene (41.2 L, 7 vol) was sparged with nitrogen for 20 minutes in a 100 L jacketed glass reactor. The reaction mixture was then heated to 70 °C. After 30 minutes, HPLC analysis indicated >99% conversion to K16. The reaction was cooled to 45 °C, treated with anhydrous dimethylsulfoxide (41.2 L, 7 vol) and triethylamine (9.42 L, 67.55 mol, 5 equiv), and heated at 65 °C. After 12 hours, HPLC analysis indicated complété consumption of K16. The reaction mixture was cooled to 0 °C and divided into two equal halves. Each half was treated with ice cold water (22 L), keeping the température below 15 °C, then extracted with ethyl acetate (2 x 20 L). The aqueous layers were extracted with ethyl acetate (18 L). The combined organic layers were washed with water (2 x 24 L), brine (24 L), dried over anhydrous sodium sulfate (2 kg) then evaporated under reduced pressure at 50 °C to give a semi-solid residue which 420 was co-evaporated with methanol (2 x 4 L) at 50 °C to give the crude product (6.65 kg) as a dark brown solid. The residue was triturated with methanol (32 L) at 65 °C for 5 hours, cooled to 15 °C over 5 hours, then filtered to give 3643 g of K17. This solid was triturated with a 1:2 mixture of acetone and methanol (18L) at 65 °C for 5 hours, cooled to 20 °C over 5 hours, then filtered. The filter cake was rinsed with a 1:2 mixture of acetone and methanol (2x3 L), followed by methanol (3 L) at 20 °C. The product was dried under nitrogen convection at 20 °C for 18 hours to give 2780 g of K17 (45.8% yield).

Step 5. A solution of7V-[(lR,2R)-2-amino-l,2-diphenyl-ethyl]-4-methylbenzenesulfonamide (22.1 g, 0.06 mol, 0.01 equiv) and dichloro-(l,2,3,4,5pentamethylcyclopenta-2,4-dien-l-yl)rhodium dimer (18.26 g, 0.03 mol, 0.005 equiv) in acetonitrile (12 L) was stirred for 30 minutes at 20 °C, then cooled to -5 °C. This solution was added to a suspension of K17 (2704 g, 6.024 mol, 1 equiv) in acetonitrile (16 L) and a mixture of formic acid (1.25 L, 33.13 mol, 5.5 equiv) and triethylamine (1.85 L, 13.25 mol, 2.2 equiv) (premixed and precooled to 0 °C) at 0 °C. The resulting mixture was stirred at 0 °C and the progress of the reaction was monitored by HP LC. After 31 hours, HPLC analysis indicated >99.9% conversion to K18. The reaction mixture was diluted with a solution of sodium bicarbonate (2.1 kg) in water (30 L). The resulting mixture was stirred for 15 minutes at 10 °C, then warmed to 15 °C and diluted with methyl /eri-butyl ether (12 L). The resulting mixture was stirred for 15 minutes at 15 °C. The layers were separated and the aqueous layer was extracted with methyl terAbutyl ether (12 L). The combined organic layers were sequentially washed with IN HCl (2 x 11 L) and brine (2 x 11 L). During the second brine wash, the pH of the brine layer was adjusted to ~8 using solid sodium bicarbonate (288 g). The organic layer was dried over anhydrous sodium sulfate (2 kg) and evaporated under reduced pressure to give 3.4 kg of crude K18. A solution of crude K18 in methyl tert-butyl ether (35 L) was treated with SiliaMetS DMT (1.7 kg) at 20 °C for 18 hours, then filtered. The filter cake was rinsed with methyl /eri-butyl ether (5 L). The combined filtrâtes were treated with SiliaMetS DMT (1.7 kg, 0.5 vol) in 3 consecutive runs at 50 °C for 5 hours. The mixture was cooled to 20 °C in between treatments and filtered. The filtrate after final trituration was evaporated at 45 °C under reduced pressure to give 2.4 kg of Kl8 (68.9% yield).

Step 6. A solution of K18 (1785.8 g (corrected for NMR purity), 3.96 mol) in methanol (12.5 L, 7 vol) at 20 °C was treated with 6 N sodium hydroxide (5.0 L, 29.71 mol, 7.5 equiv, precooled to 5 °C) added in 4 equal portions over 20 minutes in a 100 L jacketed glass reactor. The resulting solution was stirred at 40 °C. After 1.5 hours, LC-MS analysis indicated >99.9% conversion. The reaction mixture was cooled to 5-10 °C and adjusted to pH 10 to 11 with 6 N

421

HCl (4 L). The reaction mixture was partially evaporated under reduced pressure at 37 °C to remove methanol. The mixture was diluted with isopropyl acetate (18 L) and water (2 L). The resulting suspension was heated to 46 °C to give clear phases. After stirring for 15 minutes at 46 °C, the layers were separated. The aqueous layer was extracted with isopropyl acetate (10 L) at 40 °C. The combined organic layers were washed with half saturated brine (10 L), followed by water (5 L) at 40 °C. The organic layer was evaporated under reduced pressure at 40 °C to dryness to give 1298 g of crude Compound 174 (Compound 174) (~92% yield).

Step 7. Compound 174 (1207 g, 3.4 mol (corrected for -90% ’HNMR purity), 1 equiv) was co-evaporated with methyl ethyl ketone (4 L) at 40 °C under reduced pressure. The residue was dissolved in methyl ethyl ketone (6 L) and filtered (~8 pm porosity). The filtrate was charged to the reactor along with water (40 mL, 2.2 mol, 0.65 equiv). The resulting solution was heated to 60-62 °C. n-Heptane (6 L, 5 vol) was charged to the hot solution over an hour, maintaining the température at 60-62 °C. The resulting mixture was seeded (1 g, -0.1% wt) and heated at 62 °C for an hour. The resulting solution was cooled to 20 °C over 5 hours. After stirring at 20 °C for 18 hours, the suspension was filtered through Whatman #113 filter paper at 20 °C. The filter cake was washed with a 4:1 mixture of n-heptane and methyl ethyl ketone (3 L) in 2 equal portions. The product was dried under nitrogen convection at 20 °C for 3 hours to give 1091.5 g of Compound 174 Hemihydrate (Compound 174.0.5 H2O) as a white powder (86.1% yield).

Préparation 1 of Compound 174 Phosphate Hemihydrate

628 mg Compound 174 Hemihydrate was weighed in a 10 mL vial, followed by adding about 7.6 mL 2-MeTHF. About 3.7 mL 0.5 Μ H3PO4, pre-formulated via mixing with about 0.42 mL 6 Μ H3PO.4 (aq.) and about 4.6 mL MeOH was added to the vial dropwise. The mixture was stirred with a magnetic stirring bar at ambient température for two days. Then the solids were collected via centrifugation and dried in 40 °C vacuum oven ovemight. Total solids recovered were 670 mg.

Préparation 2 of Compound 174 Phosphate Hemihydrate eq. of Compound 174 Hemihydrate was charged to a reactor followed by 8 vol. of 2-MeTHF. The mixture was agitated at 40 °C. The clear solution at 40 °C was seeded with 1 wt% of Compound 174 Phosphate Hemihydrate. In a separate container, 1.02 eq. of 85 wt% phosphoric acid was diluted with 0.35 vol. of water, 3 vol. of 2-MeTHF, and 0.6 vol. of acetone. This phosphoric acid solution was then added to the reactor slowly over 2 hours. The resulting 422 slurry was then cooled to 20 °C over 5 hours. The final slurry was agitated at 20 °C for not less than 2 hours then fïltered under vacuum. The resulting wet cake was washed with 3 vol. of 2-MeTHF. The wet cake was dried under vacuum with a nitrogen bleed at 50 °C to yield about 94% of Compound 174 Phosphate Hemihydrate.

XRPD

X-ray powder diffraction (XRPD) spectra were recorded at room température (25 ± 2 °C) in transmission mode using a PANalytical Empyrean System equipped with a sealed tube source and a PIXcel 3D Medipix-3 detector (Malvem PANalytical Inc, Westborough, Massachusetts) (FIG. 24, Table 33). The X-Ray generator operated at a voltage of 45 kV and a current of 40 mA with copper radiation (1.54060 Â). The powder sample was placed on a 96 well sample holder with mylar film and loaded into the instrument. The sample was scanned over the range of about 3° to about 40° 20 with a step size of 0.0131303° and 49s per step.

Table 33. Peak List from XRPD Diffractogram of Compound 174 Phosphate Hemihydrate

No.	Pos. [±0.2, °20]	Rel. Int. [%]
1	9.1	100.0
2	16.7	77.4
3	18.7	68.1
4	20.0	43.3
5	15.7	41.9
6	14.9	39.0
7	18.4	36.1
8	10.1	32.8
9	20.2	32.4
10	15.2	27.0
11	23.9	25.7
12	20.7	25.6
13	23.6	24.6
14	16.3	23.9
15	17.1	23.4
16	21.0	21.4
17	26.2	20.5
18	22.0	20.4
19	21.2	19.8
20	19.8	19.0
21	27.4	18.1
22	17.8	18.0
23	10.2	17.1

423

No.	Pos. [±0.2, °20]	Rel. Int. [%]
24	21.6	16.6
25	24.1	15.8
26	13.2	15.3
27	25.5	14.9
28	25.7	14.8
29	18.9	12.5
30	20.4	12.0
31	22.7	11.8
32	22.3	11.7
33	17.9	11.1
34	8.8	11.0
35	19.6	10.6
36	27.0	10.5
37	10.5	10.3
38	27.2	10.1

TGA

Thermal gravimétrie analysis of Compound 174 Phosphate Hemihydrate was conducted using a TA Discovery 550 TGA from TA Instrument. A sample with a weight of 1-10 mg was scanned from 25 °C to 350 °C at a heating rate of 10 °C/min with nitrogen purge. Data were collected by Thermal Advantage Q Sériés™ software and analyzed by Trios and/or Universal Analysis software (TA Instruments, New Castle, DE). The thermogram showed 2.4% weight loss from ambient température up to 150 °C (FIG. 25).

DSC

DSC of Compound 174 Phosphate Hemihydrate was conducted using a TA Discovery 550 DSC. A sample with a weight between 1-10 mg was weighed into an aluminum crimp sealed pan with a pinhole. This pan was placed in the sample position in the calorimeter cell. An empty pan was placed in the reference position. The calorimeter cell was closed and a flow of nitrogen was passed through the cell. The heating program was set to heat the sample at a heating rate of 10 °C/min to a température of 250 °C. When the run was completed, the data were analyzed by Trios and/or Universal Analysis software (TA Instruments, New Castle, DE). The thermogram showed two endothermie peaks around 123 °C and 224 °C (FIG. 26).

SSNMR

The ¹³C CPMAS of Compound 174 Phosphate Hemihydrate (FIG. 27, Table 34) was

424 acquired at 275K with 12.5 kHz spinning and using adamantane as a reference. Additionally, the ¹³C CPMAS of Compound 174 Phosphate Hemihydrate following déhydration (FIG. 28, Table 35) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference.

Table 34. Peak List from ¹³C CPMAS of Compound 174 Phosphate Hemihydrate

Peak#	Chem Shift [ppm]	Intensity [rel]
1	144.7	17.8
2	143.5	16.3
3	142.9	20.7
4	141.3	53.4
5	140.6	4.3
6	140.2	19.6
7	139.7	20.0
8	139.1	21.1
9	136.8	30.3
10	135.9	27.0
11	129.5	20.2
12	127.6	22.6
13	127.1	29.4
14	126.6	30.0
15	125.6	24.1
16	125.1	21.1
17	123.7	20.8
18	73.0	36.2
19	72.5	100.0
20	66.1	29.9
21	65.4	32.8
22	63.4	32.1
23	62.8	17.5
24	61.4	28.2
25	50.5	62.2
26	48.4	30.7
27	47.7	41.4
28	46.9	34.6
29	43.9	21.2
30	42.6	21.0
31	40.8	20.6
32	40.5	21.9
33	39.9	31.2
34	39.4	20.8

425

Peak#	Chem Shift [ppm]	Intensity [rel]
35	39.0	26.3
36	38.6	28.0
37	37.4	23.9
38	36.7	24.6
39	36.1	23.7
40	34.6	22.34
41	18.4	27.3
42	16.6	34.58
43	15.8	31.5
44	15.3	35.4

Table 35. Peak List from ¹³C CPMAS of Dehydrated Compound 174 Phosphate Hemihydrate

Peak#	Chem Shift [ppm]	Intensity [rel]
1	144.7	23.3
2	144.1	18.6
3	143.0	33.0
4	141.3	31.2
5	140.7	17.7
6	140.2	27.1
7	139.5	14.1
8	138.6	16.1
9	138.3	18.2
10	136.8	29.8
11	129.0	22.5
12	127.5	53.1
13	125.6	42.3
14	124.7	27.6
15	123.9	22.3
16	73.3	100.0
17	73.0	55.6
18	72.2	58.5
19	66.5	60.3
20	65.0	46.4
21	64.1	57.4
22	62.4	10.5
23	61.2	46.6
24	50.2	34.2
25	48.5	60.9
26	47.6	39.0

426

Peak#	Chem Shift [ppm]	Intensity [rel]
27	46.5	39.4
28	46.1	29.9
29	45.3	26.1
30	44.0	20.1
31	42.9	46.8
32	40.6	32.6
33	39.3	53.4
34	38.5	54.1
35	37.0	33.9
36	36.6	29.5
37	34.5	21.6
38	16.5	89.0

The ¹³C CPMAS of Compound 174 Phosphate Hemihydrate (FIG. 29A, Table 36A) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference. Additionally, the ^3IP CPMAS of Compound 174 Phosphate Hemihydrate following déhydration (FIG. 29B, 5 Table 36B) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference.

Table 36A. Peak List from ³¹P CPMAS of Compound 174 Phosphate Hemihydrate

Peak#	Chem Shift [ppm]	Intensity [rel]
1	3.1	100.0
2	-1.1	70.6
3	-1.8	28.0

Table 36B. Peak List from ³¹P CPMAS of Dehydrated Compound 174 Phosphate

Hemihydrate

Peak#	Chem Shift [ppm]	Intensity [rel]
1	5.6	81.9
2	4.4	100.0
3	3.2	85.0
4	3.0	96.7

Alternative Préparation of Compound 174 Hemihydrate

100 mg of amorphous Compound 174 was added to a glass vial. To this was added 0.4 mL MEK, and ail solids dissolved. 3 pL water was then added to aid the hemihydrate formation.

To this mixture was added 0.25 mL of n-Heptane directly. After stirring for 18 hours at ambient

427 température, the solids were filtered, rinsing with 1:4 MEK/n-Heptane (v/v), followed by 100% n-Heptane. The solids were collected, dried in a vacuum oven (60 °C) ovemight, and characterized.

XRPD

X-ray powder diffraction (XRPD) spectra were recorded at room température (25 ± 2 °C) in transmission mode using a PANalytical Empyrean System equipped with a sealed tube source and a PIXcel 1D Medipix-3 detector (Malvem PANalytical Inc, Westborough, Massachusetts) (FIG. 30, Table 37). The X-Ray generator operated at a voltage of 45 kV and a current of 40 mA 10 with copper radiation (1.54060 Â). The powder sample was placed on a 96-well sample holder with mylar film and loaded into the instrument. The sample was scanned over the range of about 3° to about 40° 2Θ with a step size of 0.0131303° and 49s per step.

Table 37. Peak List from XRPD Diffractogram of Compound 174 Hemihydrate (Room

Température)

No.	Pos. [±0.2, °20]	Rel. Int. [%]
1	17.1	100.0
2 ’	20.4	87.3
3	19.1	74.1
4	6.5	66.2
5	5.7	46.8
6	14.4	32.6
7	12.1	25.6
8	11.4	22.3
9	25.5	22.0
10	12.3	20.5
11	18.9	19.9
12	9.4	19.9
13	22.4	19.7
14	21.8	18.8
15	15.8	17.7
16	22.7	14.5
17	22.4	14.2

428

No.	Pos. [±0.2, °20]	Rel. Int. [%]
18	20.8	12.9
19	25.0	12.4
20	26.1	12.1
21	29.0	12.0
22	26.1	11.8
23	27.0	11.8
24	19.3	11.5
25	25.1	11.2
26	25.3	11.1
27	6.2	10.9
28	27.9	10.9
29	28.4	10.7
30	15.7	10.0

Additionally, in one test of in situ variable température XRPD (VT-XRPD), Compound 174 Hemihydrate was observed to show peak shifts at elevated température. Variable température X-ray powder diffraction (VT-XRPD) spectra were recorded in 30-90 °C in reflection mode using a PANalytical Empyrean System equipped with a sealed tube source and a PIXcel 1D Medipix-2 detector (Malvem PANalytical Inc, Westborough, Massachusetts). The step-wise température change in incréments of 10 °C from 30 °C to 90 °C with a hold at each température for 1 hour, followed by XRD collection. The sample chamber was purged with house nitrogen. The X-Ray generator operated at a voltage of 45 kV and a current of 40 mA with 10 copper radiation (1.54060 Â). The sample was scanned over the range of about 3° to about 40°

2Θ with a step size of 0.0131303° and 49.725s per step.

Three distinct XRPD patterns were found, respectively, at: (1) ambient température to 30 °C; (2) 40-50 °C; and (3) 60-90 °C. The sample retumed to its initial form after reequilibration at ambient température and humidity. The XRPD spectrum from ambient température to 30 °C was the same (within ± 0.2 °20) as the XRPD spectrum collected at room température (25 ± 2 °C). Table 38 lists the peaks observed between 40-50 °C, and Table 39 lists the peaks observed between 60-90 °C.

429

Table 38. Peak List from XRPD Diffractogram of Compound 174 Hemihydrate (40-50 °C)

No.	Pos. [±0.2, °20]	Rel. Int. [%]
1	20.1	100.0
2	19.0	85.2
3	11.3	58.1
4	5.6	56.8
5	22.3	56.2
6	25.1	45.7
7	24.8	43.1
8	27.8	42.5
9	22.1	40.3
10	17.2	32.5
11	9.5	30.8
12	11.9	29.1
13	18.7	28.0
14	15.6	23.2
15	20.9	22.6
16	6.6	22.5
17	21.9	21.6
18	23.9	21.4
19	22.6	21.2
20	29.9	20.6
21	19.6	20.1
22	30.0	19.8
23	26.6	19.7
24	25.9	19.5
25	28.9	18.4
26	26.2	17.1
27	26.9	16.3
28	27.0	16.2
29	28.7	15.6
30	19.3	15.3
31	28.3	14.5

430

No.	Pos. [±0.2, °20]	Rel. Int. [%]
32	14.4	12.4
33	17.8	12.3
34	25.5	11.6
35	23.4	11.0
36	23.1	11.0
37	29.4	10.9
38	24.2	10.6

Table 39. Peak List from XRPD Diffractogram of Compound 174 Hemihydrate (60-90 °C)

No.	Pos. [±0.2, °20]	Rel. Int. [%]
1	19.8	100.0
2	19.2	72.1
3	5.5	62.0
4	21.8	60.5
5	11.0	50.3
6	27.2	48.3
7	24.7	46.0
8	19.0	44.5
9	22.0	40.6
10	24.3	40.4
11	17.3	35.5
12	11.7	28.8
13	9.6	28.7
14	29.3	24.2
15	29.3	23.4
16	21.0	21.8
17	26.8	20.7
18	23.5	20.5
19	15.5	20.0
20	6.7	19.9
21	27.4	18.0

431

No.	Pos. [±0.2, °20]	Rel. Int. [%]
22	25.1	17.7
23	25.8	16.4
24	23.0	15.8
25	29.9	15.1
26	14.4	14.8
27	25.9	14.4
28	28.3	14.3
29	17.8	13.2
30	15.6	13.0
31	25.6	12.3
32	20.3	12.3
33	22.6	12.2

TGA

Thermal gravimétrie analysis of Compound 174 Hemihydrate was conducted using a TA Discovery 550 TGA from TA Instrument. A sample with a weight of approximately 1-10 mg was scanned from 25 °C to 300 °C at a heating rate of 10 °C/min with nitrogen purge. Data were collected by Thermal Advantage Q SeriesTM software and analyzed by Trios and/or Universal Analysis software (TA Instruments, New Castle, DE). The thermogram showed 2.4% weight loss from ambient température up to 150°C (FIG. 31).

DSC

DSC of Compound 174 Hemihydrate was conducted using the TA Instruments Q2000 DSC. A sample with a weight between 1-10 mg was weighed into an aluminum crimp sealed pan with a pinhole. This pan was placed in the sample position in the calorimeter cell. An empty pan was placed in the reference position. The calorimeter cell was closed and a flow of nitrogen was passed through the cell. The heating program was set to heat the sample at a heating rate of 10 °C/min to a température of 300° C. When the run was completed, the data were analyzed by Trios and/or Universal Analysis software (TA Instruments, New Castle, DE). The thermogram showed endothermie peaks around 77 °C, 107 °C, and 125 °C (FIG. 32).

SSNMR

The ^l3C CPMAS of Compound 174 Hemihydrate (FIG. 33, Table 40) was acquired at

432

275K with 12.5 kHz spinning and using adamantane as a reference. Additionally, the ^I3C

CPMAS of Compound 174 Hemihydrate following déhydration (weekend at ambient température and ovemight at 80 °C in rotor) (FIG. 34, Table 41) was acquired at 275K with 12.5 kHz spinning and using adamantane as a reference.

Table 40. Peak List from ¹³C CPMAS of Compound 174 Hemihydrate

Peak#	Chem Shift [ppm]	Intensity [rel]
1	150.9	32.6
2	147.6	38.4
3	142.7	44.5
4	140.9	47.7
5	139.8	65.2
6	133.2	56.1
7	131.9	25.5
8	124.7	42.9
9	124.2	40.7
10	123.4	27.4
11	74.6	100.0
12	67.9	58.3
13	65.0	48.6
14	64.0	39.3
15	61.9	44.1
16	49.7	48.6
17	49.4	39.6
18	48.2	38.3
19	47.4	34.0
20	46.4	42.4
21	43.9	33.0
22	43.2	28.9
23	40.3	35.4
24	38.4	41.0
25	35.8	36.3

433

Peak#	Chem Shift [ppm]	Intensity [rel]
26	22.6	70.0
27	21.9	61.5

Table 41. Peak List from ^,3C CPMAS of Dehydrated Compound 174 Hemihydrate

Peak#	Chem Shift [ppm]	Intensity [rel]
1	151.9	20.5
2	147.2	17.3
3	142.2	23.6
4	141.5	25.6
5	139.4	34.1
6	132.9	29.6
7	130.7	14.4
8	125.5	21.1
9	124.4	24.0
10	121.1	17.5
11	74.6	100.0
12	67.6	39.4
13	64.1	58.3
14	61.8	30.9
15	49.9	24.8
16	49.2	25.8
17	47.7	42.9
18	47.3	39.9
19	46.6	30.3
20	45.2	26.8
21	44.3	33.9
22	39.8	26.2
23	38.5	26.3
24	35.3	22.2
25	22.6	41.1
26	22.4	43.6

434

Example 3. Assays for Detecting and Measuring APOL1 Inhibitor Properties of Compounds MultiTox-Fluor Multiplex Cytotoxicity Assay

The MultiTox-Fluor Multiplex Cytotoxicity Assay is a single-reagent-addition, homogeneous, fluorescence assay that measures the number of live and dead cells simultaneously in culture wells. The assay measures cell viability and cytotoxicity by detecting two distinct protease activities. The live-cell protease activity is restricted to intact viable cells and is measured using a fluorogenic, cell-permeant peptide glycyl-phenylalanylamino fluorocoumarin (GF-AFC) substrate. The substrate enters intact cells, where it is cleaved to generate a fluorescent signal proportional to the number of living cells. This live-cell protease activity marker becomes inactive upon loss of membrane integrity and leakage into the surrounding culture medium. A second, cell-impermeant, fluorogenic peptide substrate (bis-AAF-Rl 10 Substrate) is used to measure dead-cell protease that has been released from cells that hâve lost membrane integrity. A ratio of dead to live cells is used to normalize data.

Briefly, the tet-inducible transgenic APOL1 T-REx-HEK293 cell lines were incubated with 50 ng/mL tet to induce APOL1 in the presence of 3-(2-(4-fluorophenyl)-lH-indol-3-yl)-N((3S,4R)-4-hydroxy-2-oxopyrrolidin-3-yl)propenamide at 10.03, 3.24, 1.13, 0.356, 0.129, 0.042, 0.129, 0.0045, 0.0015, 0.0005 pM in duplicate for 24 hours in a humidified 37°C incubator. The MultiTox reagent was added to each well and placed back in the incubator for an additional 30 minutes. The plate was read on the EnVision plate reader. A ratio of dead to live cells was used to normalize, and data was imported, analyzed and fit using Genedata Screener (Basel, Switzerland) software. Data was normalized using percent of control, no tet (100% viability), and 50 ng/mL tet treated (0% viability),and fit using Smart Fit. The reagents, methods, and complété protocol for the MultiTox assay are described below.

Table 42. Reagents Used in the Multi-Tox Assay

Reagent	Catalog Number	Vendor
384 well, transparent, fiat bottom tissue culture treated, Poly-D lysine coated	356663	Corning (Corning, NY)
384 well round bottom polypropylene plates	3656	CoStar (Corning, NY)
Universal plate lids	250002	Thermo Fisher (Waltham)
Axygen 30 pL tips for Bravo 384 well	VT-384-31UL-R-S	Corning (Corning, NY)

435

Reagent	Catalog Number	Vendor
MultiTox-Fluor Multiplex Cytotoxicity Assay	G9202	Promega (Madison, WI)
225 cm² flask, angled neck, treated, vented cap	431082	Corning (Corning, NY)
Dulbecco's PhosphateBuffered Saline (DPBS), calcium and magnesiumfree	14190-136	Thermo Fisher (Waltham)
Dulbecco's Modified Eagle Medium (DMEM), high glucose, no glutamine, no sodium pyruvate	11960-077	Thermo Fisher (Waltham)
Fêtai Bovine Sérum (FBS), tetracycline-free, USSourced	631368	Takara (Kusatsu, Japan)
L-Glutamine, 200 mM	25030-081	Thermo Fisher (Waltham)
Penicillin-Streptomycin, 10,000 Units/mL	15140-122	Thermo Fisher (Waltham)
Blasticidin S HCl, 10 mg/mL	Al 1139-03	Thermo Fisher (Waltham)
Tétracycline hydrochloride	T7660 -5G	Sigma (St. Louis, MO)
Puromycin dihydrochloride, 10 mg/mL	A11138-03	Thermo Fisher (Waltham)
Trypsin-EDTA	25300-054	Thermo Fisher

(Waltham)

Table 43. Equipment Used in the Multi-Tox Assay

Instrument	Model	Supplier	Location
Bravo	16050-101	Agilent	Santa Clara, CA
		Technologies
Multidrop	N/A	Thermo	Waltham, MA
Combi		Scientific
EnVision	N/A	PerkinElmer	Waltham, MA

Multi-Tox Assay Protocol

Human embryonic kidney (HEK293) cell lines containing a tet-inducible expression

System (T-REx™; Invitrogen, Carlsbad, CA) and Adeno-associated virus site 1 pAAVSl-Puro-APOLl GO or pAAVSl-Puro-APOLl G1 or pAAVSl-Puro-APOLl G2 Clones GO DC2.13, G1 DC3.25, and G2 DC4.44 were grown in a T-225 flask at ~90% confluency in cell growth media (DMEM, 10% Tet-free FBS, 2 mM L-glutamine, 100 Units/mL penicillinstreptomycin, 5 pg/mL blasticidin S HCl, 1 pg/mL puromycin dihydrochloride). Cells were washed with DPBS and then trypsinized to dissociate from the flask. Media was used to quench the trypsin, cells were then pelleted at 200g and resuspended in fresh cell assay media (DMEM,

436

2% Tet-free FBS, 2 mM L-glutamine, 100 Units/mL penicillin-streptomycin). Cells were counted and diluted to 1.17 x 10⁶ cells/mL. 20 pL of cells (23,400/well) were dispensed to every well of a 384-well Poly-D-Lysine coated plate using the Multidrop dispenser. The plates were then incubated at room température for one hour.

Tétracycline is needed to induce APOL1 expression. 1 mg/mL tet stock in water was diluted to 250 ng/mL (5X) in cell assay media. 60 pL of cell assay media (no tet control) was dispensed in columns 1 and 24, and 60 pL of 5X tet was dispensed in 384-PP-round bottom plate in columns 2 to 23 with the Multidrop dispenser.

Assay ready plates from the Global Compound Archive were ordered using template 384_APOLlCell_DR10n2_50uM_v3. Compounds were dispensed at 200 nL in DMSO. The final top concentration was 10 pM with a 10 point 3-fold dilution in duplicate in the MultiTox assay.

pL were transferred from the 5X tet plate to the ARP and mixed, then 5 pL of 5X tet and the compounds were transferred to the cell plate and mixed using the Bravo. The cell plate was placed in the humidified 37°C 5% CO2 incubator for 24 hours.

The MultiTox-Fluor Multiplex Cytotoxicity Assay was performed in accordance with the manufacturer’s protocol. After cells were incubated with tet and compound for 24 hours, 25 pL of Ix MultiTox reagent was added to each well using the Multidrop dispenser; the plates were placed on a plate shaker (600 rpm) for 2 minutes, then centrifuged briefly and placed back in the 37°C incubator for 30 minutes. The cell viability (excitation: 400 nm, émission: 486 nm) and cytotoxicity (excitation: 485 nm, émission: 535 nm) were read using the EnVision plate reader. A ratio of dead (cytotoxicity) to live (viability) cells was reported. Data was exported and analyzed in Genedata. Data was normalized using percent of control, no tet (100% viability), and 50 ng/mL tet treated (0% viability), and fit using Smart Fit settings in Genedata.

Trypanosoma brucei brucei Lysis Assay Using APOL1 Recombinant Protein

Trypanosoma brucei brucei is a blood stream parasite to which human, gorillas, and baboons are immune due to the presence of the APOL1 protein in their HDL particles. The protein is uptaken by the parasite via the TbHpHb receptor located in its flagellar pocket and is bonded by the Hpr protein contained in the HDL particles which triggers the receptor endocytosis by the parasite.

Following endocytosis, the formed vesicle containing the HDL particle matures from early to late endosome, and subsequently to lysosome. The concomitant pH change in the lumen of the vesicle triggers the insertion of the APOL1 protein into the membrane of the late endosome/lysosome and hereby triggers lysosomal membrane permeabilization and as a further 437 downstream event, trypanosome lysis. Trypanosoma brucei brucei is sensitive to lysis by ail tbree APOL1 variants (GO, Gl, and G2).

The Trypanosoma brucei brucei lysis assay is a lysis assay of the parasite using recombinant APOL1 protein variant followed by a fluorescent détection method of viability by 5 the addition of AlamarBlue reagent to the assay well, a general metabolic redox indicator (AlamarBlue assay).

Briefly, the AlamarBlue active compound, resazurin, a blue, water soluble, non-toxic and cell permeable molécule, which can be followed by absorbance, is reduced by varions metabolic pathways into resorufin, a red compound which can be followed by either absorbance or fluorescence. The assay allows the calculation of the percent viability (percent of living Trypanosomes remaining in each well) at the end of a lysis relative to the untreated condition by interpolation of fluorescent values (FLU) on a standard curve with a known amount of seeded trypanosome/well.

Reagents and Materials

Trypanosoma brucei brucei (ATCC, Cat. No. PRA-382) o Lister 427 VSG 221 bloodstream form.

Thaw/Expansion Media (ATCC Medium 2834 Modified HMI-9 Medium)

IMDM	250 mL	76.3%
FBS	25 mL	7.63%
Sérum Plus	25 mL	7.63%
HMI-9	25 mL	7.63%
Hypoxanthine	2.5 mL 327.5 mL total	0.763%
Assay Media (No Phénol Red/No FBS): Make on Day of Use
IMDM No Phénol Red	250 mL	82.6%
Sérum Plus	25 mL	8.26%
HMI-9	25 mL	8.26%
Hypoxanthine	2.5 mL 302.5 mL total	0.826%
HMI-9 (10X)
Bathocuproine disulfonic acid	280 mg
Cysteine	1820 mg
Sodium pyruvate (lOOx)	100 mL
Uracil	100 mg
Cytosine	100 mg
2-mercaptoethanol	140 pL
Water	900 mL 1000 mL total

Hypoxanthine Stock (lOOx)-9 (10X) Sodium Hydroxide 0.8 g

438

Hypoxanthine 2.72 g

Water 200 mL

200 mL total

Table 44. Media Reagents in Trypanosoma brucei brucei used in Lysis Assay

IMDM	Phénol Red sodium pyruvate L-glutamine 25 mM HEPES	Life Technologies, Cat. No. 12440
IMDM	NO Phénol Red sodium pyruvate L-glutamine 25 mM HEPES	Life Technologies, Cat. No. 21056
FBS	Heat inactivated	Sigma-Aldrich, Cat. No. F8317-500 mL
Sérum Plus	medium supplément	Sigma-Aldrich, Cat. No. 14008C
Bathocuproine disulfonic acid		Sigma-Aldrich, Cat. No. B1125-1G
Cysteine		Sigma-Aldrich, Cat. No. C7352-25G
Sodium Pyruvate Solution	lOOx	Sigma-Aldrich, Cat. No. S8636-100ml
Uracil		Sigma-Aldrich, Çat. No. U1128-25G
Cytosine		Sigma-Aldrich, Cat. No. C3506-1G
2-mercaptoethanol		Sigma-Aldrich, Cat. No. M3148-25ml
Hypoxanthine		Sigma, Cat. No. H9636
Sodium hydroxide		Sigma-Aldrich, Cat. No. S8045-500G

Table 45. Materials used in Trypanosoma brucei brucei Lysis Assay

T75/T175	Nunc™Non-Treated flask Non-TC treated Vented/White lids with filter	T75 Thermo-Fisher Cat. No. 156800 T175 Thermo-Fisher Cat. No. 159926
Assay Plates	384 well black clear bottom Non-sterile Non-TC treated	Corning® Cat. No. 3762
Polypropylene storage plates		Corning® Cat. No. 3656
Plate Lids	Clear universal stérile lids	Thermo-Fisher Cat. No. 250002

439

Bravo Tips	30 pL tips for 384 well	Axygen Cat. No. VT-38431UL-R-S
El-Clip Tip pipette 12 channel adjustable 2-125 pL		Thermo-Fisher Cat. No. 4672070BT
Tips	125 pL El-Clip stérile filter	Thermo-Fisher Cat. No. 94420153
Tips	125 pL El-Clip stérile (nonfilter)	Thermo-Fisher Cat. No. 94410153

Equipment o El-Clip Tip pipette 12 channel adjustable 2-125 pL, Cat. No. 4672070BT o ThermoFisher MultiDrop 384, Cat. No. 5840300 o Multidrop o Agilent Bravo, Cat. No. G5409A o Bravo o SpectraMax M5

Assay Ready Plates (ARPs) o ARPs cornes in two formats:

mM final top concentration with a 2.5 fold dilution down.

mM final top concentration with a 3 fold dilution down.

Both hâve a 10 point Dose response.

0.1% DMSO final in the Black Assay Plate.

Compounds are diluted 1000 fold in the Black Assay Plate.

Each plate is designed for 14 compounds in duplicate.

o In the final Black Assay Plate:

• Column 1: Media only (no APOL1) (100% viable) • Column2-23: 0.05 pg/mL APOL1 (-ECw) (10% viable with APOL1) • Column 24: 0.1 pg/mL APOL1 (ECioo) (Approx. 0% viable)

Assay Procedures

Trypanosoma brucei brucei Culture

Protocol A

Step 1, Day 1 o Thaw the cells at 35 °C for no more than 2 minutes.

440 o Resuspend one vial gently in 20 mL pre-warmed media and incubate in a T75 flask at 37 °C and 5% CO₂.

o Do not remove the cryoprotective agent.

Step 2, Day 4 o Centrifuge at 800xg for 5 minutes at room température.

o Resuspend in 1 mL media.

o Make a 1:25 fold dilution (10 pL/240 pL media).

o Count on a hemocytometer (after adding parasites).

• Let sit for 1 -2 minutes for the parasites to settle.

• Count should be approximately 100 viable motile parasites/16 grid or approximately 25 x 10⁶ parasites/flask.

o Passage the parasites by adding 1 x 10⁶ parasites/T75 flask in 20 mL media.

o Passage the parasites by adding 2.33 x 10⁶ parasites/T175 flask in 46.6 mL media.

• For every T75 flask should make enough for approximately 1.5 x 384 well assay plates.

• For every Tl75 flask should make enough for approximately 3.8 x 384 well assay plates.

Step 3, Day 6 o Centrifuge at 800xg for 5 minutes.

• Resuspend in 3 mL assay media (No phénol red, no FBS) per 75 starting flask.

• Resuspend in 7 mL assay media (No phénol red, no FBS) per 175 flask o Make a 1:25 fold dilution.

o Count by hemocytometer.

• Every T75 flask set up should hâve approximately 75 x 10⁶ parasites/flask (verify doubling time = 8.7 hours + 1 hour).

• Every T175 flask set up should hâve approximately 175 x 10⁶parasites/flask (verify doubling time = 8.7 hours + 1 hour).

• Require 46 x 10⁶ parasites per 384 well plate (at 120,000 parasites per well).

Protocol B

Step 1, Day 1 o Thaw the cells at 35 °C for not more than 2 minutes.

441 o Resuspend one vial gently in 20 mL of pre-warmed médiate and incubate in a T75 flask at 37 °C and 5% CO₂.

o Do not remove the cryoprotective agent.

Step 2, Day 2 o Centrifuge at 800xg for 5 minutes at room température.

o Resuspend in 1 mL media.

o Make a 1:25 fold dilution (10 pL/240 μΕ media).

• Let sit for 1-2 minutes for the parasites to settle.

• Count should be approximately 100 viable motile parasites/16 grid or approximately 8 x 10⁶ parasites per flask.

o Passage the parasites by adding 1.25 x 10⁶ parasites per T75 flask in 20 mL media.

• For every T75 flask set up should hâve approximately 1.5 x 384 well assay plates.

• For every T175 flask set up should hâve approximately 3.8 x 384 well assay plates.

Step 3, Day 5 o Centrifuge at 800xg for 5 minutes.

• Resuspend in 3 mL assay media (no phénol red, no FBS) per T75 starting flask.

• Resuspend in 7 mL assay media (no phénol red, no FBS) per Tl75 starting flask.

o Make a 1:25 fold dilution.

o Count by hemocytometer.

• Every T75 flask should hâve approximately 75 x 10⁶ parasites per flask (verify doubling time: 7.7 hours + 1 hour).

• Every Tl75 flask should hâve approximately 175 x 10⁶ parasites per flask (verify doubling time: 7.7 hours + 1 hour).

Lysis Assay Setup

APOL1 G1 Protein o Remove an aliquot of the 1.2 mg/mL APOL1 protein stock from -70 °C.

o Détermine amount required for the experiment:

• Need 11.5 mL of 0.1 pg/mL APOL1 per 384 well plate.

442 • Need 0.5 mL of 0.2 pg/mL APOL1 per 384 well plate for control.

o Make initial 1:10 dilution (10 pL/90pL) into Assay media (now at 120 pg/mL).

• Using APOL1 at a final concentration of 0.05 pg/mL for an ~ECso. Need to détermine this value for each new lot of protein used.

• Adding 30 mL/well of 2X APOL1 concentration of 0.1 pg/mL.

Solution A: Measure 8.33 pL (120 pg/mL) in 10 mL for a 0.1 pg/mL 2X stock.

Solution B: Measure 16.67 pL (120 pg/mL) in 10 mL for a 0.2 pg/mL 2X stock control.

Multidrop o Black Assay Plate (384 well black well clear bottom, Cat. No. 3762).

Column 1 : Dispense 30 pL/well of Assay media (no APOL1).

Column 2-23 : Dispense 30 pL/well of Solution A (0.1 pg/mL APOL1).

Column 24: Dispense 30 pL/well of Solution B (0.2 pg/mL APOL1).

o Storage Plate (Polypropylene storage plate, Corning® Cat. No. 3656).

Column 1-24: Dispense 80 pL Assay media (no APOL1) per well (30 mL media/plate).

Bravo: Compound Transfer o Place the storage plate, the Assay Ready Plate (ARP), and Black Assay Plate on the deck.

• Transfer 20 pL from the storage plate to the ARP and mix.

• Transfer 6 pL from the ARP to the Black Assay Plate and mix.

• Black Assay Plates are now ready for Trypanosome addition.

Trypanosome Addition:

Once the Black Assay Plates hâve compounds added, begin harvesting the Trypanosomes as described in Step 3 of the Trypanosoma brucei brucei Culture section.

o Count the Trypanosomes and préparé at 5 x 10⁶/mL in Assay media (no Phénol red and no FBS).

• Requires 9.2 mL of 5 x 10⁶ trypanosomes/mL for each 384 well plate (46 x 10⁶/plate).

o Add 24 pL of 5 x 10⁶ trypanosomes mix to each well of a 384 well plate using the El-Clip multichannel 12 channel 2-125 pL adjustable pipette.

443 o Once addition is complété, tap plate on the surface to ensure liquid is within each well.

o Place plates on the plate shaker for approximately 10 seconds and shake to ensure even distribution and that no drops are left on any edges.

o Place in incubator ovemight (16 hours) at 37 °C and 5% CO2.

o Each well should include 60 pL:

pL 2X APOL1 media, 6 pL of 10X compounds, and 24 pL of trypanosome solution.

AlamarBlue Addition o After 16 hours ovemight in incubator, remove required amount of AlamarBlue (2.3 mL/plate) from the bottle stored in refrigerator, and warm up briefly in a 37 °C water bath.

o Add 6 pL/well using the El-Clip Multichannel 12 channel 2-125 pL adjustable pipette.

o Protect from light and incubate the plate at 37 °C and 5% CO2 for 2.5 hours.

o Read on SpectraMax (Softmax Pro 6.4 software, excitation: 555 nm, émission: 585 nm)

Potency Data for Compounds 1 to 390

The compounds of Formula I are useful as inhibitors of APOL1 activity. Table 46 below illustrâtes the IC50 of Compounds 1 to 390 using procedures described above. In Table 46 below, the following meanings apply. For IP50 (i.e., IC50 for cell prolifération) and IC90: “+++” means < 50 nM; “++” means between 50 nM and 500 nM; “+” means > 500 nM. N.D. = Not determined.

Table 46. Potency data for Compounds 1 to 390

Compound No.	Multi-Tox Assay	Trypanosome Assay
IC90 (nM)	IPso (nM)	Max Activity (%)	IP50 (nM)	Max Activity (%)
1	+++	+++	99.3	+++	124.1
2	+++	+++	100.0	+++	117.3
3	+++	-H-+	99.7	+++	107.0
4	+++	+++	99.3	+++	117.5
5	++	++	100.0	++	111.0
6	+++	+++	99.7	+++	102.5
7	++	++	96.0	+	92.0
8	+++	+++	100.0	++	105.5
9	++		100.0	++	108.0
10	+++	+++	99.7	++	101.0

444

Compound No.	Multi-Tox Assay	Trypanosome Assay
IC₉o(nM)	IP50 (nM)	Max Activity (%)	IPso (nM)	Max Activity (%)
11	+++	+++	99.3	+++	101.5
12	+++	H—H-	99.3	+++	102.0
13	+++	+++	99.0	+++	100.0
14	++	-H-+	99.3	+++	102.5
15	+++	+++	99.0	+++	101.0
16	++	+++	100.0	++	99.5
17	+++	+++	100.0	++	102.0
18	+++	+++	99.5	++	103.0
19	+++	+-H-	99.3	+++	106.7
20	+++	+++	99.4	+++	115.7
21	+++	+++	99.7	+++	108.0
22	+++	+++	99.7	-H-+	128.7
23	+++	+++	99.7	+++	115.7
24	+++	+++	100.0	+++	107.0
25	+++	+++	100.0	+++	109.0
26	+++	-H-+	100.0	+++	110.0
27	+++	+++	100.3	+++	112.0
28	+++	+++	100.0	+++	104.0
29	+++	+++	100.0	+++	109.0
30	+++ -	+++	99.0	+++	124.3
31	+++	+++	100.0	+++	123.0
32	+++	+++	99.6	+++	116.3
33	+++	-H-+	98.7	+++	111.0
34	+++	+++	99.0	+++	115.0
35	+++	+++	99.3	+++	111.0
36	+++	+++	100.0	+++	117.5
37	+++	+++	99.7	+++	113.5
38	+++	+-H-	100.0	+++	112.0
39	+++	+++	99.7	+++	116.0
40	+++	+++	100.0	+++	118.5
41	-H-+	+++	99.7	+++	111.5
42	H—H-	+++	99.7	+++	110.5
43	+++	+++	100.0	+++	103.0
44	+++	+++	99.5	+++	105.5
45	+++	+++	100.0	+++	103.3
46	+++	H—H-	99.0	+++	107.7
47	+++	+++	99.3	+++	100.5
48	+++	+++	99.0	+++	101.5
49	+++	+++	99.7	+++	103.5
50	+++	+++	99.7	+++	103.0

445

Compound No.	Multi-Tox Assay	Trypanosome Assay
IC90 (nM)	IP50 (nM)	Max Activity (%)	IP50 (nM)	Max Activity (%)
51	+++	+++	99.7	+++	102.5
52	+++	+++	99.7	+++	103.5
53	+++	+++	99.3	++	106.0
54	+++	+++	100.0	+++	102.5
55	+++	+++	99.7	+++	111.5
56	+++	+++	99.8	+++	115.7
57	+++	+++	99.7	+++	105.0
58	+++	+++	99.7	+++	100.5
59	+++	+++	99.3	+++	116.5
60	+++	+++	99.7	+++	102.5
61	+++	+++	99.7	+++	105.0
62	+++	+++	99.3	+++	95.0
63	+++	+++	99.8	+++	104.0
64	+++	+++	100.0	+++	101.0
65	+++	+++	99.5	+++	105.3
66	+++	+++	99.8	+++	102.7
67	+++	+++	99.7	+++	105.0
68	+++	+++	99.7	+++	105.5
69	+++	+++	99.7	+++	110.0
70	+++	+++	99.3	+++	104.0
71	+++	+++	100.0	+++	103.5
72	+++	+++	99.7	+++	118.3
73	+++	+++	99.8	+++	110.3
74	+++	+++	99.2	++	109.7
75	+++	+++	100.0	+++	105.0
76	+++	+++	99.3	+++	105.5
77	+++	+++	99.0	+++	110.0
78	+++	+++	99.7	+++	102.5
79	++	+++	99.7 .	+++	97.5
80	+++	+++	99.7	+++	100.5
81	+++	+++	99.7	+++	100.7
82	+++	+++	99.7	+++	106.3
83	+++	+++	99.0	+++	111.0
84	+++	+++	99.8	+++	112.3
85	+++	+++	99.3	+++	99.5
86	+++	+++	99.5	+++	112.0
87	+++	+++	99.3	+++	100.0
88	+++	+++	99.3	+++	98.0
89	+++	+++	99.3	+++	101.0
90	+++	+++	100.0	+++	104.5

446

Compound No.	Multi-Tox Assay	Trypanosome Assay
IC90 (nM)	IPso (nM)	Max Activity (%)	IPso (nM)	Max Activity (%)
91	+++	+++	100.0	+++	109.5
92	+++	+++	100.0	+++	103.0
93	+++	+++	99.3	++	112.0
94	+++	+++	100.0	+++	104.0
95	+++	+-H-	99.8	++	106.3
96	+++	-H-+	99.8	+++	104.0
97	+++	+++	99.0	+++	104.5
98	+++	+++	99.0	+++	110.5
99	+++	+++	99.0	+++	102.0
100	d—H-	+++	99.3	+++	99.0
101	+++	+++	100.0	++	114.3
102	+-H-	+++	99.8	+++	112.0
103	+++	+++	99.6	++	117.7
104	++	+++	99.3	++	100.5
105	++	+++	99.3	+++	72.0
106	+++	+++	99.7	+++	107.0
107	+++	+++	99.7	+++	92.5
108	++	+++	99.0	+++	94.5
109	++	+++	100.0	++	100.0
110	++	4-++	99.2	++	115.0
111	++	-H—h	99.3	++	98.5
112	+++	+++	100.0	++	112.0
113	++	+++	100.0	++	97.0
114	++	+++	99.7	++	95.0
115	++	+++	99.3	++	100.0
116	++	+++	99.4	++	100.7
117	++	+++	99.0	+++	100.7
118	++	+++	99.3	++	98.0
119	++	+++	99.7	++	97.5
120	++	+++	99.0	++	91.5
121	++	+++	99.0	++	97.5
122	++	-H-+	99.7	++	105.5
123	++	-H-+	100.0	+++	103.5
124	++	++	99.3	++	103.5
125	-H-	+++	99.7	++	103.0
126	++	++	99.4	++	114.7
127	++	++	99.3	++	107.0
128	++	++	99.3	++	107.0
129	++	-H-	100.0	+++	88.0
130	++	-H-	99.0	++	101.5

447

Compound No.	Multi-Tox Assay	Trypanosome Assay
IC₉o(nM)	IP50 (nM)	Max Activity (%)	IP50 (nM)	Max Activity (%)
131	++	++	99.3	++	111.0
132	++	++	99.3	++	91.5
133	++	++	99.7	++	111.0
134	-H-	++	99.2	++	96.7
135	++	++	98.0	++	94.0
136	-H-	++	98.7	++	100.3
137	++	++	98.7	++	84.5
138	++	++	98.0	++	104.5
139	++	++	98.0	+	98.3
140	++	++	99.7	++	101.5
141	++	+4-	96.3	++	90.0
142	-H-	-H-	99.0	++	83.7
143	-H-	++	98.0	++	101.0
144	+	++	98.0	++	101.5
145	+	++	97.7	++	88.0
146	++	++	97.0	++	88.0
147	+	++	92.3	++	93.5
148	++	++	97.0	++	92.5
149	+	++	95.7	+	95.0
150	+	++	96.3	+	87.0
151	+	++	96.2	+	85.7
152	+	++	98.0	+	92.7
153	+	++	96.0	+	89.5
154	+	+	94.0	+	96.5
155	+	-H-	95.2	+	97.0
156	+	+	93.0	+	85.0
157	+	-H-	96.3	+	95.0
158	+	+	90.4	+	75.5
159	+	+	92.2	+	63.5
160	+	+	94.3	+	97.5
161	+	+	92.4	+	73.0
162	+	+	92.0	+	81.0
163	+	+	92.7	+	79.5
164	+	+	91.7	+	85.0
165	+	+	92.0	+	61.0
166	+	+	96.7	+	57.5
167	+	+	89.7	+	73.0
168	+	+	89.0	+ '	54.5
169	+	+	86.8	+	61.0
170	+	+	89.7	+	71.5

448

Compound No.	Multi-Tox Assay	Trypanosome Assay
IC₉o(nM)	IP50 (nM)	Max Activity (%)	IP50 (nM)	Max Activity (%)
171	+	+	71.8	+	14.0
172	+	+	52.0	+	64.5
173	+++	4-4-4-	99.5	+++	116.0
174	+++	4-4-4-	99.8	+++	104.2
175	+4-+	+++	100.0	+++	105.5
176	+++	+++	100.0	+++	115.5
177	+++	+++	100.0	++	107.3
178	+++	4-4-4-	99.7	+++	101.0
179	+++	4--H-	99.0	+++	119.5
180	+++	4-4-4-	99.8	+++	102.5
181	+++	+++	99.8	+++	101.9
182	+++	4-H-	100.0	+++	113.0
183	+++	4-4-4-	99.5	++	105.5
184	++	+++	99.3	++	100.5
185	+++	+++	99.5	++	99.0
186	++	+4-4-	99.3	++	104.0
187	+4-+	+++	100.0	++	106.5
188	++	++	99.0	++	108.0
189	+	++	97.0	+	87.5
190	-H-	++	99.0	+	92.5
191	-H-	++	99.7	++	108.0
192	++	++	99.0	++ _;	100.5
193	++	++	99.0	++	100.5
194	++	++	98.7	++	117.3
195	+	+	76.0	+	50.7
196	++	-H-	99.0	++	111.0
197	-H-	++	99.0	++	106.5
198	++	++	98.7	++	99.3
199	++	+++	99.3	++	110.3
200	+	++	97.0	++	98.7
201	+	++	95.0	+	76.7
202	+	+	94.7	+	19.0
203	+++	4-H-	99.3	+++	102.3
204	4—F	++	99.0	++	112.0
205	4-4-	+++	99.3	++	105.7
206	++	4-H-	99.3	++	104.0
207	++	+++	100.0	+++	108.0
208	++	+++	99.5	++	103.3
209	+	+	76.7	++	100.3
210	+	+	92.2	+	103.5

449

Compound No.	Multi-Tox Assay	Trypanosome Assay
IC90 (nM)	IP50 (nM)	Max Activity (%)	IPso (nM)	Max Activity (%)
211	++	+++	99.6	++	113.3
212	+++	-H-+	99.0	+++	101.3
213	++	++	99.5	++	98.7
214	++	+++	99.3	++	108.7
215	++	+++	99.3	++	112.5
216	+	++	97.7	++	103.7
217	+++	+++	99.2	+++	110.4
218	++	+++	99.3	++	111.0
219	+	+	88.7	+	71.0
220	+++	+++	99.3	+++	110.0
221	++	+++	99.0
222	++	++	100.0
223	+	+	93.0
224	++	++	99.0
225	++	++	97.0
226	++	++	100.0
227	++	++	100.0
228	++	++	100.0
229	++	++	98.0
230	+	++	98.0
231	++	++	95.0
232	++	+++	100.0
233	++	++	100.0
234	++	+++	100.0
235	++	+++	100.0
236	-H-	+++	100.0
237	++	+++	99.0
238	++	++	100
239	++	++	97.0
240	+	++	95.0
241	++	+++	97.0
242	++	++	97.0
243	++	++	98.0
244	++	++	100.0
245	++	+++	99.0
246	++	++	97.0
247	+	++	91.0
248	++	+++	101.0
249	+	++	92.0
250	++	+++	95.0

450

Compound No.	Multi-Tox Assay	Trypanosome Assay
IC₉o(nM)	IP50 (nM)	Max Activity (%)	IPso (nM)	Max Activity (%)
251	++	++	99.0
252	++	+++	100.0
253	++	+++	100.0
254	+	+	92.0
255	++	++	100.0
256	++	++	96.0
257	++	-H—l·	101.0
258	+	++	98.0
259	++	++	100.0
260	++	+++	99.0
261	++	++	100.0
262	++	++	99.0
263	+	+	86.0
264	++	+++	100.0
265	++	++	99.0
266	++	+++	100.0
267	++	++	99.0
268	++	++	99.0
269	++	++	99.0
270	+	++	93.0
271	++	+++	97.0
272	+	++	95.0
273	+4-	+++	101.0
274	++	++	97.0
275	+	++	94.0
276	++	++	100.0
277	++	+-H-	100.0
278	++	+++	100.0
279	++	++	97.0
280	++	++	99.0
281	++	++	99.0
282	++	+++	99.0
283	++	++	101.0
284	++	++	101.0
285	++	++	98.0
286	++	+++	98.0
287	++	++	96.0
288	++	++	97.0
289	+	+	83.0
290	++	++	99.0

451

Compound No.	Multi-Tox Assay	Trypanosome Assay
IC90 (nM)	IPso (nM)	Max Activity (%)	IPso (nM)	Max Activity (%)
291	++	++	100.0
292	++	++	99.0
293	+++	+++	100.0
294	++	++	100.0
295	++	+++	99.0
296	++	++	99.0
297	++	+++	99.0
298	+	++	95.0
299	++	+++	99.0
300	++	++	100.0
301	+++	+++	99.0
302	++	++	94.0
303	++	++	99.0
304	++	++	99.0
305	++	+++	100.0
306	+	+	99.0
307	++	+++	99.0
308	+	++	95.0
309	++	++	97.0
310	+	++	94.0
311	++	-H-+	99.0
312	+	+	91.0
313	++	++	98.0
314	+++	+++	99.0
315	++	++	98.0
316	+	++	95.0
317	++	++	98.0
318	+++	+++	99.0
319	++	-H-	95.0
320	+		95.0
321	++	+++	99.0
322	-H-	++	97.0
323	++	+++	100.0
324	+	++	96.0
325	++	+++	98.0
326	++	++	97.0
327	++	+++	98.0
328	++	+++	100.0
329	+	+	90.0
330	++	+++	99.0

452

Compound No.	Multi-Tox Assay	Trypanosome Assay
IC90 (nM)	IPso (nM)	Max Activity (%)	IP50 (nM)	Max Activity (%)
331	++	+-H-	98.0
332	++	++	96.0
333	++	++	97.0
334	+	++	96.0
335	+	++	96.0
336	++	+++	99.0
337	++	+++	99.0
338	+++	+++	99.0
339	-H-	++	97.0
340	-H-	++	98.0
341	+	++	98.0
342	++	++	98.0
343	++	++	97.0
344	++	+++	99.0
345	++	+++	99.0
346	++	++	99.0
347	-H-	-H-	96.0
348	+	++	97.0
349	+	++	91.0
350	++	+++	99.0
351	++	+++	98.0
352	++	+++	99.0
353	+++	+++	100.0
354	++	++	99.0
355	++	++	98.0
356	++	++	99.0
357	++	++	99.0
358	+++	+++	98.0
359	+++	+++	98.0
360	++	++	99.0
361	++	H—H-	98.0
362	++	++	98.0
363	++	-H-+	99.0
364	+	++	95.0
365	++	+++	99.0
366	+	++	95.0
367	+++	+++	99.0
368	+	++	96.0
369	+	-H-	98.0
370	++	++	99.0

453

Compound No.	Multi-Tox Assay	Trypanosome Assay
IC90 (nM)	IP50 (nM)	Max Activity (%)	IP₅o (nM)	Max Activity (%)
371	-H-	+++	99.0
372	+	+	92.0
373	-H-	+++	99.0
374	++	-H-+	99.0
375	++	H—H-	99.0
376	++	++	99.0
377	+	++	98.0
378	++	+++	99.0
379	+	+	94.0
380	-H-	+++	99.0
381	+++	+++	100.0
382	++	+++	100.0
383	+++	+++	100.0
384	+++	+++	100.0
385	-H-+	+++	100.0	+++	103.5
386	+++	H—H-	100.0
387	+	+	94.0
388	+	+	90.0
389	+	++	99.0
390	++4-	+++	99.0

Other Embodiments

This disclosure pro vides merely exemplary embodiments of the disclosed subject matter.

One skilled in the art will readily recognize from the disclosure and daims, that varions changes, 5 modifications and variations can be made therein without departing from the spirit and scope of the disclosure as defined in the following daims.

Claims

1. A compound represented by the following structural formula:

<R^3a)k

5 . Formula I a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, wherein:

X¹ is selected from S and -CR^2a and X² is selected from S and -CR^2b, wherein: one of X¹ and X² is S;

10 when X¹ is S, then X² is -CR^2b; and ' ' when X² is S, then X¹ is -CR^2a;

R¹ is selected from hydrogen, halogen, cyano, -OH, Cj-Cô alkyl, Ci-Cg alkoxy, C3-C6 cycloalkyl, and phenyl, wherein:

the Ci-Cô alkyl of R¹ is optionally substituted with 1 to 3 groups independently

15 selected from halogen, cyano, -OH, -NH2, -NH(Ci-C₄ alkyl), -N(Ci-C₄ alkyl)2, and Cj-C₄ alkoxy;

. the C3-C6 cycloalkyl of R¹ is optionally substituted with 1 to 3 groups

20 ' independently selected from halogen, cyano, -OH, -NH₂, -NH(Ci-C₄ alkyl), -N(C]-C₄alkyl)₂, C_rC₄ alkyl, Ci-C₄ alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂; and the phenyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH₂, -NH(C_rC₄ alkyl), -N(C_rC₄ alkyl)₂, Ç_rC₄

25 alkyl, C_rC₄ alkoxy, -C(=O)NH₂, -C(=O)NH(C_rC₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂;

R^2ais selected from hydrogen, halogen, cyano, -OH, =0, and Çj-Ce alkyl, wherein: ' 455 the Cj-Cô alkyl of R^2a is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and C1-C4 alkoxy;

R²¹’ is selected from hydrogen, halogen, cyano, -OH, =0, and C|-C₆ alkyl;

R^3a is selected from halogen, cyano, -OH, C|-Cg alkyl, and =0; wherein:

the C|-Cô alkyl of R^3a is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

R^3bis selected from C1-C2 alkyl and =0; wherein:

------, for each occurrence, is a single bond when R^3a is selected from halogen, cyano, -OH, Ci-Cô alkyl or when R^3bis selected from C1-C2 alkyl; or»altematively------, for each occurrence, is a double bond when R^3a is =0 or when R^3b is =0;

R⁴ is selected from Cj-Cg alkyl, -C(=O)O(C]-C4 alkyl), C₂-Cg alkynyl, and

the Ci-Cô alkyl of R⁴ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(C]-C4 alkyl), -N(Ci-C4 alkyfh, C1-C4 alkoxy, -C(=O)NH₂, -C(=O)NH(Ci-C₄ alkyl), -C(=O)N(Ci-C₄ alkyl)₂, C₃-C₆ cycloalkyl, 5 to 10-membered heterocyclyl, phenyl, and 5 to 10-membered heteroaryl;

Ring A is selected from C3-C12 carbocyclyl, 3 to 12-membered heterocyclyl, Cg and Cio aryl, and 5 to 10-membered heteroaryl, wherein Ring A is optionally substituted with 1, 2, 3, 4, or 5 R^a groups; wherein:

R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cô alkyl, C2-C6 alkenyl, C]-Cg alkoxy, C]-C6 haloalkyl, Cj-Cg haloalkenyl, Ci-C6 haloalkoxy, -C(=0)NR^hRⁱ, -NR^hR^!, -NR^hC(=0)R^k, -NR^hC(=0)0R^k, -NR^hC(=0)NR'R^j, -NR^hS(=O)pR^k-OR^k, -OC(=O)R^k, -OC(=O)OR^k, -0C(=0)NR^hRⁱ, -[O(CH₂)_q]_rO(Ci-C₆ alkyl), -S(=O)_pR^k, -8(=0)^^, -C(=O)OR^k, C3-C12 carbocyclyl, 3- to 12-membered heterocyclyl, Cg and Cio aryl, and 5- to 10-membered heteroaryl; wherein:

the Ci-Cg alkyl, Ci-Cg alkoxy, and the C2-C0 alkenyl of R^a are each optionally substituted with 1 to 3 groups independently selected

456 from C₆ to Cio aryl (optionally substituted with 1 to 3 Rⁿⁱ groups), 5- to 10-membered heterocyclyl (optionally substituted with 1 to 3 R”' groups), 5- to 10-membered heteroaryl (optionally substituted with 1 to j R groups), cyano, -C(=O)R^k, -C(=O)OR^k, -C(=O)NR R, -NR R, -NRC(=O)R^k, -NRC(=O)OR^k, -NR^llC(=O)NRⁱR^j, -NR^hS(=O)pR^k. -OR^k, -OC(=O)R^k, -OC(=O)OR^k, -OC(=O)NR^hRⁱ, -S(=O)pR^k, -8(=0)^^^, and C₃-C₆ carbocyclyl (optionally substituted with 1 to 3 R' groups);

the C3-C12 carbocyclyl, the 3 to 12-membered heterocyclyl, the C₆ and Cio aryl, and the 5 to 10-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups independently selected-from halogen, cyano, C1-C4 alkyl, -NR^hR‘, and -OR^k; wherein:

R^h, R‘, and R^j, for each occurrence, are each independently selected from hydrogen, C1-C4 alkyl, C₆-Cio aryl, and C₃-C₆ cycloalkyl; wherein:

the C1-C4 alkyl of any one of R^h, R¹, and R^j is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, and -OH;

R^k, for each occurrence, are each independently selected from hydrogen, C1-C4 alkyl, 5- to 10-membered heterocyclyl, and C₃-C₆ carbocyclyl; wherein:

R^m, for each occurrence, is independently selected from halogen, cyano, oxo, Ci-Cô alkyl, Ci-Cg alkoxy, -S(=O)_PR , and -OR^k; wherein:

R^sis selected from Ci-C₆ alkyl, -C(=O)O(CrC₄ alkyl), C₃-Ci₂ carbocyclyl, 3- to 12membered heterocyclyl, Cô and Cio aryl, and 5- to 10-membered heteroaryl, wherein.

the Ci-Cô alkyl of R⁵ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH₂, -NH(Ci-C4 alkyl), -N(Cj-C4 alkyl)₂, C1-C4

457 alkoxy. *

-C(=O)NH₂, -C(=O)NH(C>-C₄ alkyl), and -C(=O)N(Ci-C₄ alkyl)₂;

the C3-C12 carbocyclyl, the 3 to 12-membered heterocyclyl, the C₄₎ and Cioaryl, and the 5 to 10-membered heteroaryl of R^? are each optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH₂, -NH(Cj-C₄ alkyl) (optionally substituted with -OH), -N(Ci-C₄ alkyl)₂, C1-C5 alkyl (optionally substituted with -OH), Cj-C₄ alkoxy, -C(=O)NH₂, -C(=O)NH(C,-C₄ alkyl), -NHC(=O)(CrC₄ alkyl), -0(=0)(0)-C₄ alkoxy), and -C(=0)N(C,-C₄ alkyl)₂;

k is an integer selected from 0, 1, and 2, wherein:

when R^3a is selected from halogen, cyano, -OH, and C]-Cô alkyl, k is 1 or 2; and when R^3a is =0, k is 1 ; , .

m is an integer selected from 0, 1, and 2, wherein:

when R^3b is selected from C)-C₂ alkyl, m is 1 or 2; and when R^3b is =0, m is 1 ;

2. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to claim 1, wherein the compound is represented by one of the following structural formulae:

or is a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing, wherein:

R^2ais selected from hydrogen, halogen, cyano, and C)-C₄ alkyl; wherein:

45¾ the C|-Cj allÇyl of R²ⁱⁱ is optionally substituted with 1 to 3 groups independently selected from halogen, -OH, and C1-C2 alkoxy; '

R^2b is selected from hydrogen, halogen, cyano, and C1-C4 alkyl; and k is an integer selected from 0, 1, and 2.

The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait (R^a)₀-5

according to claim 1 or claim 2, wherein R⁴ is selected from C1-C4 alkyl and » wherein:

the C,-C₄ alkyl of R⁴ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, -NH2, -NH(Ci-C4 alkyl), -N(Ci-C₄ alkyl)2, C1-C2alkoxy, C3-C6 cycloalkyl, 5 to 6-membered heterocyclyl, phenyl, and 5 to 6-membered heteroaryl.

4. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 3, wherein R⁴ is selected from C1-C2 alkyl and

the C1-C2 alkyl of R⁴is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and 5 to 6-membered heterocyclyl.

5. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 4, wherein R⁴ is selected from -CH3, -CH2OH, and (tetrahydro-2H-pyran-4-yl)methyl.

6. The compound according to any one of daims 1 to 4, wherein the compound is represented by one of the following structural formulae:

453

5 Ring A, for each occurrence, is selected from C3-C6 cycloalkyl, 5- to 10-membered heterocyclyl, phenyl, and 5- to 10-membered heteroaryl; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups.

7. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait

10 according to any one of daims 1 to 4 and 6, wherein Ring A is selected from cyclopropyl, 5- to 10-membered heterocyclyl, phenyl, and 5- to 9-membered heteroaryl; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups.

8. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait

15 according to any one of daims 1 to 4, 6, and 7, wherein Ring A is selected from cyclopropyl, 5to 10-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, phenyl, and 5- to 9-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups.

20 9. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 4 and 6 to 8, wherein Ring A is selected from cyclopropyl, 5-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 6-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 9-membered heterocyclyl containing 1 to 3 heteroatoms selected from N and O, 10-membered heterocyclyl containing 1 to

25 3 heteroatoms selected from N and O, phenyl, 5-membered heteroaryl containing 1 to 3

469 heteroatoms selected from N and O. 6-membered heteroaryl contai ning 1 to 3 heteroatoms selected from N and O, and 9-membered heteroaryl containing 1 to 3 heteroatoms selected from N and O; each of which is optionally substituted with 1,2, 3, 4, or 5 R^a groups.

; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a groups.

461

462.

5 12. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 4 and 6 to 11, wherein R⁴ is selected from -CH3 and Ring A;

and

; each of which is optionally substituted with 1, 2, 3, 4, or 5 R^a

10 groups.

13. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 12, wherein R^s is selected from C1-C4 alkyl, -C(=O)O(Ci-C2 alkyl), C3-C6 cycloalkyl, and 5- to 10-membered heterocyclyl; wherein:

15 the C1-C4 alkyl of R⁵ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and C1-C2 alkoxy; and

463 the C3-C6 cycloalkyTand the 5 lo 10-membered heterocyclyl of R^are each optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, C]-C₂ alkyl, and C1-C2 alkoxy.

5

14. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 13, wherein R⁵ is selected from C|-C₂ alkyl, C(=O)O(Ci-C₂alkyl), cyclopropyl, cyclobutyl, and 5- to 6-membered heterocyclyl; wherein:

the C1-C2 alkyl of R'¹ is optionally substituted with 1 to 3 groups independently selected from F, Cl, Br, cyano, -OH, and Ci-C₂ alkoxy; and

10 the cyclopropyl, the cyclobutyl, and the 5- to 6-membered heterocyclyl of R⁵ are each optionally substituted with 1 to 3 groups independently selected fron) F, Cl, Br, cyano,. -OH, C1-C2 alkyl, and C1-C2 alkoxy.

15. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait

15 according to any one of daims 1 to 14, wherein R⁵ is selected from -CH3, -CH2CH3, -CH2OH, -C(=O)OCH3, -CH2OCH3, -CH(CH3)₂, cyclopropyl, difluorocyclopropyl, and tetrahydro-2Hpyranyl.

16. The compound according to any one of daims 1 to 15, wherein the compound is

20 represented by one of the foliowing structural formulae:

(R^3a)k (R^3a)k

Formula IVa

Formula IVb or is a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing.

464

17. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 16, wherein R¹ is selected from hydrogen, halogen, cyano, -OH, C1-C4 alkyl, C1-C4 alkoxy, and C₃-C₆ cycloalkyl; wherein:

the C1-C4 alkoxy of R¹ is optionally substituted with 1 to 3 independently selected halogen groups; and the C₃-C₆ cycloalkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, cyano, -OH, and C1-C2 alkoxy.

18. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 17, wherein R¹ is selected from F, Cl, Br, C1-C4 alkyl, and C₃-Cô cycloalkyl; wherein:

the C1-C4 alkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen and -OH; and the C₃-Cô cycloalkyl of R¹ is optionally substituted with 1 to 3 groups independently selected from halogen, and -OH.

Z

19. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 18, wherein R¹ is selected from F, Cl, Br, C1-C4 alkyl, and C₃-Cg cycloalkyl; wherein:

20. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 16, wherein R¹ is selected from Cl, Br, -CH₃, -CF₃, -CH₂CH₃, -CH(CH₃)₂, -CH2CHF2, -CH₂CH(CH₃)₂, difluorocyclobutyl, and cyclohexyl.

21. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 20, wherein R¹ is Cl.

46-5

22. The compound. tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 21. wherein R^3il is selected from halogen, -OH, and C|-C₄alkyl; wherein:

the C|-C₄ alkyl of R^3ais optionally substituted with 1 to 3 groups independently selected from halogen and -OH.

23. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 22, wherein R^3a is selected from F, Cl, Br, -OH, and Cj-Cj alkyl; wherein:

the C1-C2 alkyl of R^3a is optionally substituted with 1 to 3 groups independently selected from F, Cl, and -OH. , .

24. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 23, wherein R^3a is selected from F, -OH, -CH3, -CHF2, and CH₂OH.

25. The compound according to any one of daims 1 to 4 and 6 to 24, wherein the compound is represented by one of the following structural formulae:

Formula Va

Formula Vb or is a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing.

26. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 4 and 6 to 25, wherein R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cg alkyl, Cj-C4 alkoxy, Ci-Cô haloalkyl, Cj-Cô haloalkoxy, -C(=O)NR^hRⁱ, -NR^hR', -NR^hC(=O)R^k, -OR^k, -[O(CH2)_q]_rO(Ci-C₆ alkyl),

466

-S(=O)₂R^k, -S(=O)2NR^llRⁱT Ca-Cô cycloalkyl, 5- to 10-membered heterocyclyl, phenyl, and 5- to 8-membered heteroaryl; wherein:

the C|-C₆ alkyl of R^a is optionally substituted with 1 to 3 groups independently selected from cyano, -C(=O)NR¹’Rⁱ, -NR'W, -NR^hC(=O)R^k, -NR^hC(=O)OR^k, -ΝΗ^ηΟ(=Ο)ΝΚ^, -NR^hS(=O)pR^k -OR^k, -S(=O)2R^k, -S(=O)pNR^hR‘, and C3-C₆ cycloalkyl;

the C₃-C₆ cycloalkyl, the 5 to 10-membered heterocyclyl, the phenyl, and the 5- to 8membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups independently selected from halogen, Cj-C₂ alkyl, and -OR^k; wherein:

R¹¹, R', and R^j, for each occurrence, are each independently selected from hydrogen, C]-C₂ alkyl, cyclopropyl, and cyclobutyl; wherein:

the C,-C₂ alkyl of any one of R^h, R‘, and R^j is optionally substituted with 1 to 3 groups independently selected from halogen and -OH;

R^k, for each occurrence, are each independently selected from hydrogen and

C1-C4 alkyl; wherein:

the C1-C4 alkyl of R^k is optionally substituted with l'to 3 groups independently selected from halogen and -OH; and q and r are each an integer selected from 1, 2, and 3.

Z

27. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 4 and 6 to 26, wherein R^a, for each occurrence, is independently selected from halogen, cyano, Ci-Cg alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, CiC4 haloalkoxy, -Ο(=Ο)ΝΚ^ηΚ, -NR^hR', -NR^hC(=O)R^k, -OR^k, -[O(CH2)_q]_rO(Ci-C₄ alkyl), S(=O)₂R^k, -S(=O)₂NR^llRⁱ, cyclopropyl, cyclobutyl, 5 to 6-membered heterocyclyl, phenyl, and 5 to 6-membered heteroaryl; wherein:

the Ci-C₆ alkyl of R^a is optionally substituted with 1 to 3 groups independently selected from cyano, -C(=O)NR^hR', -NR^hR', -OR^k, cyclopropyl, and cyclobutyl;

the cyclopropyl, the cyclobutyl, the 5 to 6-membered heterocyclyl, the phenyl, and the 5 to 6-membered heteroaryl of R^a are each optionally substituted with 1 to 3 groups independently selected from halogen, -CH₃, -OH, and -OCH₃; wherein:

R^h and R¹, for each occurrence, are each independently selected from hydrogen, -CH₃, cyclopropyl, and cyclobutyl; wherein:

the -CH₃ of any one of R^h and R' is optionally substituted with 1 to 3 groups independently selected from F, Cl, and -OH;

467

R^k, for each occurrence, is each independently selected from hydrogen and -CH₃; wherein:

the -CH₃of R^k is optionally substituted with 1 to 3 groups independently selected from halogen and -OH.

28. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 4 and 6 to 27, wherein R^a, for each occurrence, is independently selected from F, Cl, Br, cyano, Ci-Cô alkyl, Cj-C2 alkoxy, Cj-C2 haloalkyl, -C(=O)NR^hR', -NR^hR', -NR^hC(=O)R^k, -OR^k, -[O(CH2)_q]_rO(C]-C₂ alkyl), -S(=O)₂R^k, -S(=O)2NR^ilR^l, cyclopropyl, cyclobutyl, 5-membered heterocyclyl, phenyl, and 6-membered heteroaryl; wherein: , .

the Ci-Cô alkyl of R^a is optionally substituted with 1 to 3 groups independently selected from cyano, -C(=O)NR^hR', -OR^k, and cyclopropyl;

R¹¹ and R¹, for each occurrence, are each independently selected from hydrogen, -CH₃, and cyclopropyl; wherein: .

Z the -CH₃ of any one of R^h and R¹ is optionally substituted with 1 to 3 groups independently selected from F, Cl, and -OH;

R^k, for each occurrence, is each independently selected from hydrogen and -CH₃; and q and r are each an integer selected from 1 and 2.

29. The compound, tautomer, deuterated dérivative, or pharmaceutically acceptable sait according to any one of daims 1 to 4 and 6 to 28, wherein R^a, for each occurrence, is independently selected from F, cyano, -OH, -CH₃, -CF₃, -CH(CH₃)₂, -(CH₂)₂OH, -(CH₂)₂OCH₃, -CH₂CH(OH)C₂H₅, -CH₂C(CH₃)(CH₂OH)₂, -och₃, -OCH₂CH₃, -[O(CH₂)₂]₂OCH₃, -CH₂C(=O)NHCH₃, -(CH₂)₂SO₂CH₃, -CH₂C(=O)N(CH₃)₂, -CH₂(cyclopropyl), -C(=O)NH₂, -C(=O)NH(cyclopropyl), -NH₂, -NHCH₃, -N(CH₃)₂, -NHC(CH₃)₂CH₂OH, -NHC(=O)CH₃, -SO₂CH₃, -SO₂NH₂, cyclopropyl, 2-methoxyphenyl, N-methylpiperazinyl, tetrahydro-2H-pyranyl, methylpyrazolyl, pyridinyl, and tetrahydrothiophenyl 1,1-dioxide.

46?

Formula Va’

Formula Vb’

5 or is a tautomer thereof, a deuterated dérivative of that compound or tautomer, or a pharmaceutically acceptable sait of any of the foregoing.

474

472

47J

475

476

477

47¾

47«

484

4SI

483

Ο /

48^

48ί

48f

OH

483

-¾

48?

489

49ί

491

tautomers thereof, deuterated dérivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.

5 32. A pharmaceutical composition comprising at least one entity according to any one of daims 1 to 31 and a pharmaceutically acceptable carrier.

33. An entity according to any one of daims 1 to 31 or a pharmaceutical composition according to daim 32 for use in method of treating focal segmentai glomerulosclerosis and/or 10 non-diabetic kidney disease in a patient.

34. An entity according to any one of daims 1 to 31 or a pharmaceutical composition according to daim 32 for use in a method of inhibiting APOL1 activity.

15

35. A Silicon dérivative of the at least one entity according to any one of daims 1 to 31.

36. A pharmaceutical composition comprising a Silicon dérivative according to daim 35.

493

37. A Silicon dérivative according lo daim 35 or a pharmaceutical composition according to daim 36 for use in a method of treating focal segmentai glomerulosclerosis and/or nondiabetic kidney disease in a patient.

38. A boron dérivative of the at least one entity according to any one of daims 1 to 31.

39. A pharmaceutical composition comprising a boron dérivative according to daim 38.

40. A boron dérivative according to daim 38 or a pharmaceutical composition according to daim 39 for use in a method of treating focal segmentai glomerulosclerosis and/or nondiabetic kidney disease in a patient.

41. A phosphorus dérivative of at least one entity according to any one of daims 1 to 31.

42. A pharmaceutical composition comprising a phosphorus dérivative according to daim

41.

S

43. A phosphorus dérivative according to daim 41 or a pharmaceutical composition according to daim 42 for use in a method of treating focal segmentai glomerulosclerosis and/or non-diabetic kidney disease in a patient.

44. An entity according to any one of daims 1 to 31 or a pharmaceutical composition according to daim 32 for use in a method of treating an APOLl-mediated disease in a patient.

45. The entity or composition according to daim 44, wherein the APOLl-mediated disease is cancer.

46. The entity or composition according to daim 44 or daim 45, wherein the APOLlmediated disease is pancreatic cancer.

47. The entity or composition according to daim 44, wherein the APOLl-mediated disease is an APOLl-mediated kidney disease.