CN117771378A

CN117771378A - Pharmaceutical composition for improving bioavailability of KRAS inhibitor and application thereof

Info

Publication number: CN117771378A
Application number: CN202311249600.7A
Authority: CN
Inventors: 吴颢; 路渊; 杨翔; 徐人奇; 杨晓峰; 李波燕; 夏洪峰; 何将旗; 孙佳玲; 匡翠文; 赵志昌; 湛波; 王维; 田凯; 李树森; 章凯帆; 张洪波; 林远望; 高锜; 方龙城
Original assignee: Betta Pharmaceuticals Co Ltd
Current assignee: Betta Pharmaceuticals Co Ltd
Priority date: 2022-09-29
Filing date: 2023-09-26
Publication date: 2024-03-29

Abstract

The invention belongs to the field of medicines, and particularly relates to a pharmaceutical composition of a KRAS inhibitor and a PGY1 inhibitor, and application of the pharmaceutical composition in preparation of oral cancer drugs. The PGY1 inhibitor and the KRAS inhibitor are combined, so that the bioavailability of the KRAS inhibitor can be greatly improved, the PK property of the KRAS inhibitor is remarkably improved, and the KRAS inhibitor can be prepared into oral medicines.

Description

Pharmaceutical composition for improving bioavailability of KRAS inhibitor and application thereof

Technical Field

The invention belongs to the field of medicines, and particularly relates to a pharmaceutical composition of a KRAS inhibitor and a PGY1 inhibitor and application of the pharmaceutical composition in preparation of oral cancer drugs.

Background

RAS is the first oncogene identified in human tumors and one of the most widely occurring oncogenes. There have been difficulties in developing targeted drugs that inhibit RAS-driven cancers, and are therefore also referred to as "non-patentable" targets. Members of the RAS gene family currently known include KRAS, NRAS and HRAS. Among them, the first oncogene KRAS mutation found in human cancer in 1982 is most common, accounting for about 85%. The KRAS mutation type and probability vary depending on the tumor tissue type, and most often carry missense mutations, including single amino acid substitutions such as glycine 12 (G12), glycine 13 (G13), and glutamine 61 (Q61).

Scientists design a covalent small molecule inhibitor capable of irreversibly targeting and combining cysteine residues on KRAS No. 12 codons by virtue of the characteristic that cysteine newly introduced by KRAS G12C mutation is easy to form covalent bonds, and finally realize the marketing of medicines. KRAS G12D differs from KRAS G12C only in the kind of amino acid mutation at the same codon. KRAS G12D is the introduction of mutation at codon KRAS12 into aspartic acid, rather than cysteine, and is directly expressed as an epidemiological difference. It is generally believed that KRAS mutations occur at about 30% in malignant tumors, although the ratio varies among tumor species. KRAS occurs in the highest proportion in pancreatic cancer, which is also commonly known as "king in cancer", and has limited therapeutic approaches per se. This "point" differs from KRAS G12C, which focuses the dominant field of KRAS G12D on pancreatic cancer, as opposed to KRAS G12C, which focuses mainly on lung cancer. Unlike the incidence of KRAS G12C in the lung cancer area, KRAS G12D is also an order of magnitude higher in pancreatic cancer, further illustrating the urgency of developing KRAS G12D inhibitors.

At present, KRAS inhibitors in clinical and research stages have poor oral bioavailability and are difficult to prepare oral medicines.

Disclosure of Invention

The inventor finds that the PGY1 inhibitor and the KRAS inhibitor are combined in the research and development process of KRAS small molecule inhibition, so that the oral bioavailability of the KRAS inhibitor can be greatly improved, the PK properties (including the whole blood exposure of the KRAS inhibitor in animals, the oral bioavailability and the like) of the KRAS inhibitor in the mice and rats are remarkably improved, and the oral tumor inhibition effect of the KRAS inhibitor can be greatly improved by combining the PGY1 inhibitor and the KRAS inhibitor through a CDX tumor-bearing mouse model of a KRAS mutant cell line, and the KRAS inhibitor concentration in the tumor tissues of the mice is remarkably improved after the PGY1 inhibitor is combined through the PK is verified through investigation.

The PGY1 inhibitor provided by the invention is a P-glycinate 1 inhibitor, which is also called a PGP inhibitor. The PGP inhibitor in the present invention means an active molecule which can play a role in reducing the expression of P-glycoprotein or inhibiting the activity of P-glycoprotein, and includes, but is not limited to, a molecule which has been currently marketed or clinically used as a PGP inhibitor, as long as it can play a role in reducing the expression of P-glycoprotein or inhibiting the activity of P-glycoprotein.

In one aspect, the present application provides a pharmaceutical combination comprising: (i) KRAS inhibitors; and (ii) a PGY1 inhibitor.

In some embodiments of the invention, the KRAS inhibitor is selected from a compound of the following general formula (I), or a stereoisomer, tautomer, deuterate, or pharmaceutically acceptable salt thereof:

wherein,

x is selected from bond, O, -NR _a -, -5-14 membered heterocyclyl-or-C _2-4 Alkynyl-; wherein the-5-14 membered heterocyclic group-, -C _2-4 Alkynyl-optionally further substituted with one or more R _a Substituted;

l is selected from the group consisting of bond, -cycloalkyl (preferably C) _3-14 Cycloalkyl) -, C _0-3 Alkylene-, -C _0-3 Alkylene-cycloalkyl (preferably C _3-14 Cycloalkyl) -or-C _0-3 Alkylene-cycloalkyl (preferably C _3-14 Cycloalkyl) -C _0-3 An alkylene group; wherein said-cycloalkyl (preferably C _3-14 Cycloalkyl) -, C _0-3 Alkylene-, -C _0-3 Alkylene-cycloalkyl (preferably C _3-14 Cycloalkyl) -, C _0-3 Alkylene-cycloalkyl (preferably C _3-14 Cycloalkyl) -C _0-3 Alkylene-optionally further substituted with one or more R _a Substituted; and when two R _a When the same carbon atom is substituted, two R _a Together with the carbon atoms to which they are attached may form C _3-14 Cycloalkyl, 3-14 membered heterocyclyl;

R ₁ selected from H, -C _0-3 alkylene-N (R) _a ) ₂ 、-C _0-3 Alkylene-heterocyclyl (preferably 3-14 membered heterocyclyl), and,-C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 alkylene-C _6-18 Aryl or-C _0-3 Alkylene-5-18 membered heteroaryl; wherein the-C _0-3 alkylene-N (R) _a ) ₂ 、-C _0-3 Alkylene-heterocyclyl (preferably 3-14 membered heterocyclyl), -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 alkylene-C _6-18 Aryl, -C _0-3 Alkylene-5-18 membered heteroaryl optionally further substituted with one or more R _a Substituted; and when two R _a When the same atom is substituted, two R _a Together with the atoms to which they are attached may form C _3-14 Cycloalkyl, 3-14 membered heterocyclyl;

d is selected from CR ₅ 、C(R ₅ ) ₂ O, N or NR ₆ ；

E is selected from C, CH or N;

f is selected from C, CH, O or N;

independently selected from single bond or double bond;

k is an N-containing heterocycle (preferably a 3-14 membered N-containing heterocycle); wherein the N-containing heterocycle (preferably 3-14 membered N-containing heterocycle) is optionally further substituted with one or more R _a Substituted;

R ₂ selected from the group consisting of absence, C _3-14 Cycloalkyl, 3-14 membered heterocyclyl, C _6-18 Aryl or 5-18 membered heteroaryl; wherein the C _3-14 Cycloalkyl, 3-14 membered heterocyclyl, C _6-18 Aryl, 5-18 membered heteroaryl optionally further substituted with one or more R _a Substitution;

R ₃ selected from H, hydroxy, halogen, cyano, C _1-6 Alkyl, C _1-6 Alkoxy or C _3-14 cycloalkyl-O-; wherein the C _1-6 Alkyl, C _1-6 Alkoxy, C _3-14 cycloalkyl-O-optionally further substituted with one or more R _a Substitution;

optionally R ₃ And substituent R on K _a Together with the atoms to which they are attached form a 6-8 membered heterocyclic ringA base; wherein the 6-8 membered heterocyclyl is optionally further substituted with one or more R _a Substituted;

R ₄ selected from the group consisting of absent, H, halogen, oxo, C _1-6 Alkoxy, C _1-6 Haloalkoxy, -O-C _0-3 alkylene-C _3-14 Cycloalkyl, -O-C _0-3 Alkylene-3-14 membered heterocyclyl, -O-C _0-3 alkylene-C _6-18 Aryl or-O-C _0-3 Alkylene-5-18 membered heteroaryl; wherein the-O-C _0-3 alkylene-C _3-14 Cycloalkyl, -O-C _0-3 Alkylene-3-14 membered heterocyclyl, -O-C _0-3 alkylene-C _6-18 Aryl or-O-C _0-3 Alkylene-5-18 membered heteroaryl optionally further substituted with one or more R _a Substitution;

R ₅ selected from H, hydroxy, oxo, halogen, cyano, C _1-6 Alkyl, C _1-6 Haloalkyl, -S-C _1-6 Alkyl, -C _0-3 alkylene-C _2-4 Alkenyl-, C _1-6 Alkoxy, C _1-6 Haloalkoxy or C _3-14 Cycloalkyl;

R ₆ selected from H, hydroxy, halogen, cyano, C _1-6 Alkyl, C _1-6 Haloalkyl, -S-C _1-6 Alkyl, -C _0-3 alkylene-C _2-4 Alkenyl-, C _1-6 Alkoxy, C _1-6 Haloalkoxy or C _3-14 Cycloalkyl;

R _a each independently selected from H, halogen, hydroxy, amino, oxo, nitro, cyano, carboxyl, C _2-6 Alkynyl, C _1-6 Alkyl, C _2-6 Olefins, -C _1-3 alkylene-C _2-6 Olefins, C _1-6 Hydroxyalkyl, C _1-6 Aminoalkyl, -C _1-3 Alkylene-cyano, C _1-6 Haloalkyl, -C _0-3 alkylene-C _1-6 Alkoxy radicalRadical, C _1-6 alkyl-S-, C _1-6 Haloalkoxy, C _1-6 Heteroalkyl, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl or-C _0-3 Alkylene-5-14 membered heteroaryl; wherein the C _1-6 Alkyl, C _2-6 Olefins, -C _1-3 alkylene-C _2-6 Olefins, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl, -C _0-3 Alkylene-5-14 membered heteroaryl optionally further substituted with one or more halo, hydroxy, cyano, hydroxy, amino, nitro, C _1-6 Alkyl, C _3-14 Cycloalkyl, 3-8 membered heterocyclyl, C _6-14 Aryl or 5-14 membered heteroaryl;

R _c 、R _d each independently selected from H, halogen, -C _0-3 Alkylene C _3-6 Cycloalkyl, -C _0-3 Alkylene C _6-8 Aryl, C _1-6 Alkyl or C _1-6 A haloalkyl group; alternatively, R _c And R is _d Together with the carbon atom to which they are attached form a 3-8 membered heterocyclic group or C _3-8 Cycloalkyl groups.

In some embodiments of the invention, K in formula (I) is a 6-10 membered N-containing heterocycle, wherein the 6-10 membered N-containing heterocycle is optionally substituted with one or more R _a Substituted; preferably, the 6-10 membered N-containing heterocycle is selected from Preferably, said R _a Selected from H, cyano, halogen, hydroxy, amino, nitro, C _1-6 Alkyl,/->-C _0-3 Alkylene-cyano, C _1-6 Haloalkyl or C _1-6 An alkoxy group.

In some embodiments of the invention, R in the general formula (I) ₁ Selected from the group consisting of

In some embodiments of the invention, X in the general formula (I) is selected from the group consisting of bond, -O-, -NH-, -5-14 membered heterocyclyl-or-C _2-4 Alkynyl-.

In some embodiments of the invention, L in the general formula (I) is selected from the group consisting of a bond, -cyclopropyl-, -C _0-3 Alkylene-, -C _0-3 alkylene-cyclopropyl-or-C _0-3 alkylene-cyclopropyl-C _0-3 Alkylene-.

In some embodiments of the invention, the formula (I) wherein-X-L-R ₁ Selected from the group consisting of

In some embodiments of the invention, R in the general formula (I) ₂ Selected from the group consisting of

In some embodiments of the invention, R in the general formula (I) ₃ Selected from hydrogen, halogen, C _1-6 Alkoxy, -O-CH ₂ -CF ₃ 、-O-cyclopropyl.

In some embodiments of the invention, R in the general formula (I) ₄ Selected from hydrogen, halogen, C _1-6 Haloalkoxy, oxo, C _1-6 Alkoxy, -O-C _0-3 alkylene-C _3-14 Cycloalkyl, -O-C _0-3 Alkylene-3-14 membered heterocyclyl, -O-C _0-3 alkylene-C _6-18 Aryl or-O-C _0-3 Alkylene-5-18 membered heteroaryl; wherein the-O-C _0-3 alkylene-C _3-14 Cycloalkyl, -O-C _0-3 Alkylene-3-14 membered heterocyclyl, -O-C _0-3 alkylene-C _6-18 Aryl or-O-C _0-3 Alkylene-5-18 membered heteroaryl optionally further substituted with one or more R _a Substitution, said R _a Each independently selected from H, halogen, hydroxy, amino, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl or-C _0-3 Alkylene-5-14 membered heteroaryl.

In some embodiments of the invention, R in formula (I) ₅ Selected from hydrogen, halogen, C _1-6 alkyl-S-, oxo, -C _0-3 alkylene-C _2-4 Alkenyl-, C _3-6 Cycloalkyl or C _1-6 An alkoxy group.

In some embodiments of the invention, the KRAS inhibitor is selected from a compound of formula I-A, I-B or I-C, or a stereoisomer, tautomer, deuterate, or pharmaceutically acceptable salt thereof:

wherein said K is selected fromWherein said->Optionally further by one or more R _a Substituted;

R ₃ selected from hydrogen, halogen, C _1-6 Alkoxy, -O-CH ₂ -CF ₃ -O-cyclopropyl;

optionally R ₃ And substituent R on K _a Together with the atoms to which they are attached form a 6-8 membered heterocyclic group; wherein the 6-8 membered heterocyclyl is optionally further substituted with one or more R _a Substituted;

x is selected from bond, -O-, -NR _a -, -5-14 membered heterocyclyl-or-C _2-4 Alkynyl-; wherein the-5-14 membered heterocyclic group-, -C _2-4 Alkynyl-optionally further substituted with one or more R _a Substituted;

l is selected from the group consisting of bond, -cyclopropyl-, -C _0-3 Alkylene-, -C _0-3 alkylene-cyclopropyl-or-C _0-3 alkylene-cyclopropyl-C _0-3 An alkylene group;

R ₁ selected from the group consisting of

R ₂ Selected from the group consisting of

R ₄ Selected from hydrogen, halogen, C _1-6 Alkoxy, C _1-6 Haloalkoxy, oxo, -O-C _0-3 alkylene-C _3-14 Cycloalkyl, -O-C _0-3 Alkylene-3-14 membered heterocyclyl, -O-C _0-3 alkylene-C _6-18 Aryl or-O-C _0-3 Alkylene-5-18 membered heteroaryl; wherein the-O-C _0-3 alkylene-C _3-14 Cycloalkyl, -O-C _0-3 Alkylene-3-14 membered heterocyclyl, -O-C _0-3 alkylene-C _6-18 Aryl or-O-C _0-3 Alkylene-5-18 membered heteroaryl optionally further substituted with one or more R _a Substitution;

R ₅ selected from H, hydroxy, halogen, cyano, C _1-6 Alkyl, C _1-6 Haloalkyl, -S-C _1-6 Alkyl, -C _0-3 alkylene-C _2-4 Alkenyl-, C _1-6 Alkoxy, C _1-6 Haloalkoxy or C _3-14 Cycloalkyl;

R _a selected from hydrogen, cyano, halogen, hydroxy, amino, nitro, C _1-6 Alkyl group, C _2-6 Alkynyl, -C _0-3 Alkylene-cyano, C _1-6 Haloalkyl, C _1-6 Haloalkoxy, C _1-6 Alkoxy, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl or-C _0-3 Alkylene-5-14 membered heteroaryl;

In some embodiments of the invention, the KRAS inhibitor is selected from the compounds in table 1, or stereoisomers, tautomers, deuterates, or pharmaceutically acceptable salts thereof:

TABLE 1

In some embodiments of the invention, the KRAS inhibitor is selected from a KRAS G12D inhibitor or a KRAS G12C inhibitor.

In some embodiments of the invention, the KRAS G12C inhibitor is selected from Sotorasib, adagrasib, GF-105, JDQ-443, YL-15293, D-1553, JAB-21822, ZG-19018, JMKX-001899, HS-10370, GDC-6036, BPI-421286, GH-35, RMC-6291, GEC-255, LY-3537982, MK-1084, D3S-001, HBI-2438, BI-1823911, JS-116, XNW-14010, ABSK-071, ARS-1620, APG-1842, RM-007, ERAS-3490, MRTX-1257, ERAS-3691, WDB-178, ICP-915, AZ-8037, ASP-2453, AZD-4625, AU-10458, AU-8653, ATG-012, LC-2, ARS-853, KP-14, RM-018, YF-135, YK-3720, JN-3728, or JN-36.

In some embodiments of the invention, the KRAS G12D inhibitor is selected from MRTX-1133, HRS-4642, JAB-22000, ERAS-4057, JR-6000, RMC-9805, IMC-KRAS-G12D, KAL-21404358, VRTX-144, RM-036, or KD-8.

In some embodiments of the invention, the PGY1 inhibitor is selected from Losartan, dactinomycin, levofloxacin, gamidin D, grepafloxacin, rifamycin, lamotrigine, posaconazole, cetirizine, rantidine, loratadine, ranolazine, mefloquine, quinine, chloroquine, dipyridamole, ticagrelor, acetaminophen, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, velplatasvir, voxilaprevir, pibrentasvir, glecaprevir, letermovir, tenofovir, disoproxil, mirabegron, bisoprolol, verapamil, mibefradil, nicardipine, amlodipine, diltiazem, tezacaftor, elexacaftor, taurocholic acid, bromocriptine, chlorpromazine, haloperidol, paliperidone, elagolix, enasidenib, lvosidenib, cyclosporine, erythromycin, clarithromycin, azithromycin, indomethacin, slidenafil, vardenafil, valinomycin, esomeprazole, pantoprazole, omeprazole, lansoprazole, lasmiditan, canagliflozin, venlafaxine, quinidine, citalopram, prazosin, atorvastain, methyl blue, tolvaptan, encequidar, dexverapamil, dexniguldipine, elacridar, tariquidar (XR-9576), zosuquidar, mitotane, laniquidar, valspodar, tezacaftor, piperine, timcodar dimesilate, licochalcone A, dofequidar fumarate, biricodar, cinchonine, cyclosporine (Huons), CTP-786, STI-0529, BST-204, schisandrin B, KI-1102, CBT-1, ORX-102, ORX-101, ATNX-04, WS-10, CAP-0121, WS-691, WS-898, YS-370, TTT-28, CP-778875, EMD 55900, PMI-002, HE-10, SDZ-280-446, W-198, BIBW-22, S-9788, ONT-093, CRL-1336, SDZ-280125, LYP18 or OC-10426.

In some embodiments of the invention, the PGY1 inhibitor is selected from Amlodipine, verapamil, bisoprolol, nicardipine, diltiazem, ticagrelor, dipyridamole, prazosin, quinidine, atorvastatin, canagliflozin, cyclosporine, indomethacin, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, glecaprevir, tenofovir disoproxil, quinine, chloroquine, dexverapamil, dexniguldipine, valspodar, dofequidar fumarate, zosuquidar, laniquidar, mitotane, biricodar, elacridar, ONT-093, encequidar, tezacaftor or Piperine.

In some embodiments of the present invention, the PGY1 inhibitor is selected from cyclosporin A, progesterone Progesterone, propafenone Propanone, nifedipine, felodipine, aureobasidin A, fluphenazine flufenazine, chlorpromazine chloromazine, caffeine, yohimbine, reserpine, ganciclovir, tamoxifen, tolrimafil, darunamine Daroutin, fostanner Fostamatinib, ritonavir, tacrolimus Tacrolimus, oxazolquidambaridavir, ranolazine, ionolazine, itraconazole, tiatavanavir, saquinavir; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

In some embodiments of the invention, the KRAS inhibitor is selected from the group consisting of a compound of formula (I), or a stereoisomer, tautomer, deuterate, or pharmaceutically acceptable salt thereof:

wherein,

l is selected from the group consisting of bond, -cycloalkyl (preferably C) _3-14 Cycloalkyl) -, C _0-3 Alkylene-, -C _0-3 Alkylene-cycloalkyl groups are preferably C _3-14 Cycloalkyl) -or-C _0-3 Alkylene-cycloalkyl (preferably C _3-14 Cycloalkyl) -C _0-3 An alkylene group; wherein said-cycloalkyl (preferably C _3-14 Cycloalkyl) -, C _0-3 Alkylene-, -C _0-3 Alkylene-cycloalkyl (preferably C _3-14 Cycloalkyl) -, C _0-3 Alkylene-cycloalkyl (preferably C _3-14 Cycloalkyl) -C _0-3 Alkylene-optionally further substituted with one or more R _a Substituted; and when two R _a When the same carbon atom is substituted, two R _a Together with the carbon atoms to which they are attached may form C _3-14 Cycloalkyl, 3-14 membered heterocyclyl;

R ₁ selected from H, -C _0-3 alkylene-N (R) _a ) ₂ 、-C _0-3 Alkylene-heterocyclyl (preferably 3-14 membered heterocyclyl), -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene group-C _6-18 Aryl or-C _0-3 Alkylene-5-18 membered heteroaryl; wherein the-C _0-3 alkylene-N (R) _a ) ₂ 、-C _0-3 Alkylene-heterocyclyl (preferably 3-14 membered heterocyclyl), -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 alkylene-C _6-18 Aryl, -C _0-3 Alkylene-5-18 membered heteroaryl optionally further substituted with one or more R _a Substituted; and when two R _a When the same atom is substituted, two R _a Together with the atoms to which they are attached may form C _3-14 Cycloalkyl, 3-14 membered heterocyclyl;

d is selected from CR ₅ 、C(R ₅ ) ₂ O, N or NR ₆ ；

E is selected from C, CH or N;

f is selected from C, CH, O or N;

independently selected from single bond or double bond;

k is an N-containing heterocycle (preferably a 3-14 membered N-containing heterocycle) which is further substituted with one or more R _a Substituted;

R ₅ selected from H, hydroxy, oxo, halogen, cyano, C _1-6 Alkyl, haloalkyl, -S-C _1-6 Alkyl, -C _0-3 alkylene-C _2-4 Alkenyl-, C _1-6 Alkoxy or C _3-14 Cycloalkyl;

R _a each independently selected from H, halogen, hydroxy, amino, oxo, nitro, cyano, carboxyl, C _2-6 Alkynyl, C _1-6 Alkyl, C _2-6 Olefins, -C _1-3 alkylene-C _2-6 Olefins, C _1-6 Hydroxyalkyl, C _1-6 Aminoalkyl, -C _1-3 Alkylene-cyano, C _1-6 Haloalkyl, -C _0-3 alkylene-C _1-6 Alkoxy, C _1-6 alkyl-S-, C _1-6 Haloalkoxy, C _1-6 Heteroalkyl, -C _0-3 alkylene-C _3-14 Cycloalkyl radicals、-C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl or-C _0-3 Alkylene-5-14 membered heteroaryl; wherein the C _1-6 Alkyl, C _2-6 Olefins, -C _1-3 alkylene-C _2-6 Olefins, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl, -C _0-3 Alkylene-5-14 membered heteroaryl optionally further substituted with one or more halo, hydroxy, cyano, hydroxy, amino, nitro, C _1-6 Alkyl, C _3-14 Cycloalkyl, 3-8 membered heterocyclyl, C _6-14 Aryl or 5-14 membered heteroaryl;

R _c 、R _d each independently selected from H, halogen, -C _0-3 Alkylene C _3-6 Cycloalkyl, -C _0-3 Alkylene C _6-8 Aryl, C _1-6 Alkyl or C _1-6 A haloalkyl group; alternatively, R _c And R is _d Together with the carbon atom to which they are attached form a 3-8 membered heterocyclic group or C _3-8 Cycloalkyl;

the PGY1 inhibitor is selected from Amlodipine, verapamil, bisoprolol, nicardipine, diltiazem, ticagrelor, dipyridamole, prazosin, quinidine, atorvastatin, canagliflozin, cyclosporine, indomethacin, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, glecaprevir, tenofovir disoproxil, quinine, chloroquine, dexverapamil, dexniguldipine, valspodar, dofequidar fumarate, zosuquidar, laniquidar, mitotane, biricodar, elacridar, ONT-093, encequidar, tezacaftor or Piperine.

Wherein,

R ₁ selected from H, -C _0-3 alkylene-N (R) _a ) ₂ 、-C _0-3 Alkylene-heterocyclyl (preferably 3-14 membered heterocyclyl), -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 alkylene-C _6-18 Aryl or-C _0-3 Alkylene-5-18 membered heteroaryl; wherein the-C _0-3 alkylene-N (R) _a ) ₂ 、-C _0-3 Alkylene-heterocyclyl (preferably 3-14 membered heterocyclyl), -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 alkylene-C _6-18 Aryl, -C _0-3 Alkylene-5-18 membered heteroaryl optionally further substituted with one or more R _a Substituted; and when two R _a When the same atom is substituted, two R _a Together with the atoms to which they are attached may form C _3-14 Cycloalkyl, 3-14 membered heterocyclyl;

d is selected from CR ₅ 、C(R ₅ ) ₂ O, N or NR ₆ ；

E is selected from C, CH or N;

f is selected from C, CH, O or N;

independently selected from single bond or double bond;

R _a each independently selected from H, halogen, hydroxy, amino, oxo, nitro, cyano, carboxyl, C _2-6 Alkynyl, C _1-6 Alkyl, C _2-6 Olefins, -C _1-3 alkylene-C _2-6 Olefins, C _1-6 Hydroxyalkyl, C _1-6 Aminoalkyl, -C _1-3 Alkylene-cyano, C _1-6 Haloalkyl, -C _0-3 alkylene-C _1-6 Alkoxy, C _1-6 alkyl-S-, C _1-6 Haloalkoxy, C _1-6 Heteroalkyl, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl or-C _0-3 Alkylene-5-14 membered heteroaryl; wherein the C _1-6 Alkyl, C _2-6 Olefins, -C _1-3 alkylene-C _2-6 Olefins, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl, -C _0-3 Alkylene-5-14 membered heteroaryl optionally further substituted with one or more halo, hydroxy, cyano, hydroxy, amino, nitro, C _1-6 Alkyl, C _3-14 Cycloalkyl, 3-8 membered heterocyclyl, C _6-14 Aryl or 5-14 membered heteroaryl;

R _c 、R _d each independently selected from H, halogen, -C _0-3 Alkylene C _3-6 Cycloalkyl, -C _0-3 Alkylene groupC _6-8 Aryl, C _1-6 Alkyl or C _1-6 A haloalkyl group; alternatively, R _c And R is _d Together with the carbon atom to which they are attached form a 3-8 membered heterocyclic group or C _3-8 Cycloalkyl;

the PGY1 inhibitor is selected from cyclosporine ACyclosporine A, progesterone Progesterone, propafenone Propanonene, nifedipine, felodipine, aureobasidin A, fluphenazine flufenazine, chlorpromazine chloromazine, caffeine, yohimbine, reserpine, ganciclovibrillin, tamoxifen, tolrimcontroller, daruloramide Daroutiamide, futaminib, ritonavir, tacrolimus, fluquindazole, ranolazine, itraconazole, tianavir, saquinavir; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

In some embodiments of the invention, the KRAS inhibitor is selected from a compound of formula I-a, or a stereoisomer, tautomer, deuterate, or pharmaceutically acceptable salt thereof:

wherein said K is selected fromSaid->Optionally further by one or more R _a Substituted;

optionally R ₃ And substituent R on K _a Together with the atoms to which they are attached form a 6-8 membered heterocyclic group, said 6-8 membered heterocyclic group optionally being further substituted with one or more R _a Substituted;

R ₁ selected from the group consisting of

R ₂ Selected from the group consisting of

wherein said K is selected from Said->Optionally further by one or more R _a Substituted;

R ₁ selected from the group consisting of

R ₂ Selected from the group consisting of

the PGY1 inhibitor is selected from cyclosporin A, progesterone Progesterone, propafenone Propanone, nifedipine, felodipine, aureobasidin A, fluphenazine flufenazine, chlorpromazine chloromazine, caffeine, yohimbine, reserpine, ganciclovir, tamoxifen, tolrimafil, darunamine Daroutin, fostanner Fostamatinib, ritonavir, tacrolimus Tacrolimus, oxazolquidambaridavir, ranolazine, ionolazine, itraconazole, tiatavanavir, saquinavir; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

In some embodiments of the invention, the KRAS inhibitor is selected from a compound of formula I-B, or a stereoisomer, tautomer, deuterate, or pharmaceutically acceptable salt thereof:

R ₁ selected from the group consisting of

R ₂ Selected from the group consisting of

wherein said K is selected from Wherein said-> Optionally further by one or more R _a Substituted;

R ₁ selected from the group consisting of

R ₂ Selected from the group consisting of

In some embodiments of the invention, the KRAS inhibitor is selected from a compound of formula I-C, or a stereoisomer, tautomer, deuterate, or pharmaceutically acceptable salt thereof:

R ₁ selected from the group consisting of

R ₂ Selected from the group consisting of

R _a Selected from hydrogen, cyano, halogen, hydroxy, amino,Nitro, C _1-6 Alkyl group, C _2-6 Alkynyl, -C _0-3 Alkylene-cyano, C _1-6 Haloalkyl, C _1-6 Haloalkoxy, C _1-6 Alkoxy, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl or-C _0-3 Alkylene-5-14 membered heteroaryl;

R ₁ selected from the group consisting of

R ₂ Selected from the group consisting of

the PGY1 inhibitor is selected from cyclosporin A cycloporine A, progesterone Progesterone, propafenone Propanone, nifedipine, felodipine felopirine, aureobasidin A, fluphenazine flufenazine, chlorpromazine chloromazine, caffeine, yohimbine, reserpine, ganciclovibrolamine, tamoxifen, tolrimitinib, ritonafen, tacrolimus Tacrolimus, zolidazoquidambarir, ranolazine, itraconazole, tioconazole, tiatavanavir, saquinavir; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

In some embodiments of the invention, the KRAS inhibitor is selected from a compound of table 1, or a stereoisomer, tautomer, deuterate, or pharmaceutically acceptable salt thereof; the PGY1 inhibitor is selected from Amlodipine, verapamil, bisoprolol, nicardipine, diltiazem, ticagrelor, dipyridamole, prazosin, quinidine, atorvastatin, canagliflozin, cyclosporine, indomethacin, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, glecaprevir, tenofovir disoproxil, quinine, chloroquine, dexverapamil, dexniguldipine, valspodar, dofequidar fumarate, zosuquidar, laniquidar, mitotane, biricodar, elacridar, ONT-093, encequidar, tezacaftor or Piperine.

In some embodiments of the invention, the KRAS inhibitor is selected from a compound of table 1, or a stereoisomer, tautomer, deuterate, or pharmaceutically acceptable salt thereof; the PGY1 inhibitor is selected from cyclosporine ACyclosporine A, progesterone Progesterone, propafenone Propanonene, nifedipine, felodipine, aureobasidin A, fluphenazine flufenazine, chlorpromazine chloromazine, caffeine, yohimbine, reserpine, ganciclovibrillin, tamoxifen, tolrimcontroller, daruloramide Daroutiamide, futaminib, ritonavir, tacrolimus, fluquindazole, ranolazine, itraconazole, tianavir, saquinavir; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

In some embodiments of the invention, the KRAS G12D inhibitor is selected from MRTX-1133, HRS-4642, JAB-22000, ERAS-4057, JR-6000, RMC-9805, IMC-KRAS-G12D, KAL-21404358, VRTX-144, RM-036, or KD-8; the PGY1 inhibitor is selected from Amlodipine, verapamil, bisoprolol, nicardipine, diltiazem, ticagrelor, dipyridamole, prazosin, quinidine, atorvastatin, canagliflozin, cyclosporine, indomethacin, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, glecaprevir, tenofovir disoproxil, quinine, chloroquine, dexverapamil, dexniguldipine, valspodar, dofequidar fumarate, zosuquidar, laniquidar, mitotane, biricodar, elacridar, ONT-093, encequidar, tezacaftor or Piperine.

In some embodiments of the invention, the KRAS G12D inhibitor is selected from MRTX-1133, HRS-4642, JAB-22000, ERAS-4057, JR-6000, RMC-9805, IMC-KRAS-G12D, KAL-21404358, VRTX-144, RM-036, or KD-8; the PGY1 inhibitor is selected from cyclosporin A cycloporine A, progesterone Progesterone, propafenone Propanone, nifedipine, felodipine felopirine, aureobasidin A, fluphenazine flufenazine, chlorpromazine chloromazine, caffeine, yohimbine, reserpine, ganciclovibrolamine, tamoxifen, tolrimitinib, ritonafen, tacrolimus Tacrolimus, zolidazoquidambarir, ranolazine, itraconazole, tioconazole, tiatavanavir, saquinavir; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

In some embodiments of the invention, the KRAS G12C inhibitor is selected from Sotorasib, adagrasib, GF-105, JDQ-443, YL-15293, D-1553, JAB-21822, ZG-19018, JMKX-001899, HS-10370, GDC-6036, BPI-421286, GH-35, RMC-6291, GEC-255, LY-3537982, MK-1084, D3S-001, HBI-2438, BI-1823911, JS-116, XNW-14010, ABSK-071, ARS-1620, APG-1842, RM-007, ERAS-3490, MRTX-1257, ERAS-3691, WDB-178, ICP-915, AZ-8037, ASP-2453, AZD-4625, AU-10458, AU-8653, ATG-012, LC-2, ARS-853, KP-14, RM-018, YF-135, YK-3720, JN-3728, or JN-36; the PGY1 inhibitor is selected from Amlodipine, verapamil, bisoprolol, nicardipine, diltiazem, ticagrelor, dipyridamole, prazosin, quinidine, atorvastatin, canagliflozin, cyclosporine, indomethacin, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, glecaprevir, tenofovir disoproxil, quinine, chloroquine, dexverapamil, dexniguldipine, valspodar, dofequidar fumarate, zosuquidar, laniquidar, mitotane, biricodar, elacridar, ONT-093, encequidar, tezacaftor or Piperine.

In some embodiments of the invention, the KRAS G12C inhibitor is selected from Sotorasib, adagrasib, GF-105, JDQ-443, YL-15293, D-1553, JAB-21822, ZG-19018, JMKX-001899, HS-10370, GDC-6036, BPI-421286, GH-35, RMC-6291, GEC-255, LY-3537982, MK-1084, D3S-001, HBI-2438, BI-1823911, JS-116, XNW-14010, ABSK-071, ARS-1620, APG-1842, RM-007, ERAS-3490, MRTX-1257, ERAS-3691, WDB-178, ICP-915, AZ-8037, ASP-2453, AZD-4625, AU-10458, AU-8653, ATG-012, LC-2, ARS-853, KP-14, RM-018, YF-135, YK-3720, JN-3728, or JN-36; the PGY1 inhibitor is selected from cyclosporin A, progesterone Progesterone, propafenone Propanone, nifedipine, felodipine, aureobasidin A, fluphenazine flufenazine, chlorpromazine chloromazine, caffeine, yohimbine, reserpine, ganciclovir, tamoxifen, tolrimafil, darunamine Daroutin, fostanner Fostamatinib, ritonavir, tacrolimus Tacrolimus, oxazolquidambaridavir, ranolazine, ionolazine, itraconazole, tiatavanavir, saquinavir; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

In some embodiments of the invention, the ratio of the average daily dose of KRAS inhibitor to PGY1 inhibitor is selected from 100:1 to 1:100; optionally, the average daily dose ratio is selected from 20:1 to 1:20, 1:10 to 10:1, 1:5 to 5:1, 1:4 to 4:1, 1:3 to 3:1, 1:2 to 2:1, or 1:1.

In some embodiments of the invention, the KRAS inhibitor is administered three times per day, twice per day, once every two days, once every three days, once every four days, once every five days, once every six days, once every week, once every two weeks, or once every three weeks.

In some embodiments of the invention, the KRAS inhibitor is administered at a dose of 1-5000mg per administration; preferably, the KRAS inhibitor is administered at a dose of 1-1000mg, 1-500mg, 5-500mg, 10-500mg, 50-400mg, 50-300mg, 50-200mg, 50-100mg, 100-500mg, 100-400mg, 100-300mg or 100-200mg per administration.

In some embodiments of the invention, the PGY1 inhibitor is administered three times per day, twice per day, once per two days, once per three days, once per four days, once per five days, once per six days, once per week, once per two weeks, or once every three weeks.

In some embodiments of the invention, the PGY1 inhibitor is administered at a dose of 0.01-1000mg per administration; preferably, the PGY1 inhibitor is administered at a dose of 0.1-100mg, 0.1-80mg, 0.1-50mg, 0.1-20mg, 1-100mg, 1-50mg, 5-100mg, 5-50mg, 5-30mg, 10-100mg, 10-50mg, or 10-30mg per time.

In some embodiments of the invention, the pharmaceutical combination is a fixed combination; optionally the fixed combination is in the form of a solid pharmaceutical composition; optionally, the KRAS inhibitor and PGY1 inhibitor in the fixed combination are present in the same solid pharmaceutical composition.

In some embodiments of the invention, the pharmaceutical composition is a non-fixed combination; optionally, the KRAS inhibitor and PGY1 inhibitor in the non-fixed combination are each in the form of a solid pharmaceutical composition; optionally, each of the KRAS inhibitor and PGY1 inhibitor in the non-fixed combination is in the form of a solid pharmaceutical composition, and the solid pharmaceutical composition of the KRAS inhibitor and the solid pharmaceutical composition of the PGY1 inhibitor are present in the same pouch; optionally, each of the KRAS inhibitor and PGY1 inhibitor in the non-fixed combination is in the form of a solid pharmaceutical composition, and the solid pharmaceutical composition of the KRAS inhibitor and the solid pharmaceutical composition of the PGY1 inhibitor are not present in the same pouch.

In some embodiments of the invention, the KRAS inhibitor and PGY1 inhibitor may be administered to the patient simultaneously, separately and/or sequentially.

In some embodiments of the invention, the pharmaceutical combination is an oral formulation.

In some embodiments of the invention, the pharmaceutical combination is a tablet or capsule.

The invention also provides a medicine box which comprises the medicine combination.

In a further aspect, the present invention provides the use of a pharmaceutical combination or kit as described above for the manufacture of a medicament, preferably an oral medicament, for the treatment of KRAS mediated diseases.

Further, the KRAS mediated disease, the cancer is preferably selected from breast cancer, multiple myeloma, bladder cancer, endometrial cancer, gastric cancer, cervical cancer, rhabdomyosarcoma, non-small cell lung cancer, polymorphic lung cancer, ovarian cancer, esophageal cancer, melanoma, colorectal cancer, hepatoma, head and neck tumor, hepatobiliary cell cancer, myelodysplastic syndrome, glioblastoma, prostate cancer, thyroid cancer, xu Wangshi cell tumor, lung squamous cell carcinoma, licheniform keratosis, synovial sarcoma, skin cancer, pancreatic cancer, testicular cancer, or liposarcoma.

In yet another aspect, the invention provides a kit for treating a KRAS-mediated disease comprising (i) a KRAS inhibitor, and (ii) a PGY1 inhibitor; the method is characterized in that: (i) Contained in a first compartment, and (ii) contained in a second compartment.

In addition, the present invention provides a method of treating a KRAS-mediated disease comprising co-administering to a subject in need of treatment a therapeutically effective amount of (i) a KRAS inhibitor; and (ii) a PGY1 inhibitor.

The pharmaceutical composition can obviously improve the bioavailability of the KRAS inhibitor, obviously improve the PK property of the KRAS inhibitor, and is favorable for preparing the KRAS inhibitor into an oral medicament.

The components of the pharmaceutical combination of the present invention may optionally be combined with one or more pharmaceutically acceptable carriers, wherein the components may each independently, or some or all of them together comprise pharmaceutically acceptable carriers and/or excipients.

The pharmaceutical combinations of the invention may be formulated separately from each other or some or all of them may be formulated together. Preferably, the components of the pharmaceutical combination are formulated separately or each as a suitable pharmaceutical composition. In some embodiments, the pharmaceutical combinations of the present invention may be formulated into pharmaceutical compositions suitable for single or multiple administration.

In some particular embodiments of the present invention, the pharmaceutical composition comprising a KRAS inhibitor may be selected from solid pharmaceutical compositions including, but not limited to, tablets or capsules.

The components of the pharmaceutical combination of the present invention may each be administered alone or some or all of them may be co-administered. The components of the pharmaceutical combinations of the present application may be administered substantially simultaneously, or some or all of them may be administered substantially simultaneously.

The components of the pharmaceutical combinations of the present invention may each be administered independently, or some or all of them together, by suitable routes, including, but not limited to, oral or parenteral.

In some embodiments of the invention, the components of the pharmaceutical combinations of the present application may each be administered independently, or some or all of them may be co-orally.

The components of the pharmaceutical combinations of the present invention may each independently, or some or all of them together, be in a suitable dosage form, including, but not limited to, tablets, troches, pills, capsules (e.g., hard capsules, soft capsules, enteric capsules, microcapsules), elixirs, granules, syrups, granules, emulsions, suspensions, solutions, dispersions, and sustained release formulations for oral administration.

In the present invention, the components of the pharmaceutical combination may be administered to a patient as separate entities (e.g., pharmaceutical compositions) simultaneously, alternately or sequentially, wherein the active ingredients administered to the patient are at therapeutically effective levels.

In some embodiments of the invention, the individual active components may be packaged, marketed or administered as a completely separate pharmaceutical composition. The amount of each component administered in the pharmaceutical combination of the present invention may be determined based on the severity of the disease, the response of the disease, any treatment-related toxicity, the age and health of the patient.

For the purposes of this application, the following terms, as used in the specification and claims, shall have the following meanings, unless otherwise indicated.

"patient" means a mammal, preferably a human.

By "pharmaceutically acceptable" is meant that it is used to prepare a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and includes that it is acceptable for human pharmaceutical use.

By "therapeutically effective amount" is meant an amount of a compound that, when administered to a human being for treating a disease, is sufficient to effect treatment of the disease.

"treatment" means any administration of a therapeutically effective amount of a compound and includes:

(1) Inhibiting the disease in a human experiencing or exhibiting the pathology or symptomology of the disease (i.e., arresting further development of the pathology and/or symptomology), or

(2) Improving the disease in a human experiencing or exhibiting the pathology or symptomology of the disease (i.e., reversing the pathology and/or symptomology).

"optionally" means with or without.

Unless otherwise indicated, general chemical terms used in the structural formulae have their ordinary meanings.

For example, the term "halogen" as used herein refers to fluorine, chlorine, bromine or iodine unless otherwise indicated.

In the present invention, unless otherwise indicated, "alkyl" includes straight or branched monovalent saturated hydrocarbon groups. For example, alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 3- (2-methyl) butyl, 2-pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-methylpentyl and the like. Similarly, "C _1-6 "in" alkyl group " _1-6 "refers to a group comprising an array of straight or branched chain forms of 1, 2, 3, 4, 5 or 6 carbon atoms.

"alkoxy" refers to the oxyether form of the aforementioned straight or branched alkyl group, i.e., -O-alkyl.

The term "alkylene" refers to a divalent alkyl linking group. Alkylene refers formally to an alkane in which two C-H bonds are replaced with points of attachment of the alkylene group to the rest of the compound. Similarly, C _1-3 "C" in alkylene _1-3 "refers to an alkylene group containing 1,2 or 3 carbon atoms and includes, but is not limited to, methylene, 1, 2-ethylene, 1, 3-propylene or 1, 2-isopropylene.

The term "haloalkyl" refers to an alkyl group in which one or more H has been replaced with a halogen atom.

The term "oxo" or "oxo" refers to an oxygen atom in the form of a divalent substituent that forms a carbonyl group when attached to C and a sulfoxide or sulfone group or an N-oxide group when attached to a heteroatom.

The term "aryl", in the present invention, unless otherwise indicated, refers to an unsubstituted or substituted monocyclic or fused ring aromatic group comprising atoms of a carbocyclic ring. Preferably C _6-18 Aryl, more preferably aryl is C _6-10 A monocyclic or bicyclic aromatic ring group. Preferably phenyl, naphthyl; most preferred is naphthyl. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl group, wherein the ring attached to the parent structure is an aryl ring, non-limiting examples of which include, but are not limited to, benzocyclopentyl.

The term "heterocyclyl" refers to a ring system having at least one cyclized alkyl or cyclized alkenyl group containing a heteroatom selected from N, O and/or S. The heterocyclyl may include single or multiple rings (e.g., having 2, 3, or 4 fused rings, spiro rings, bridged rings, etc.). The heterocyclic group may be attached to the rest of the compound via a ring-forming carbon atom or a ring-forming heteroatom. Preferably a 3-14 membered heterocyclic group, and "3-14 membered" in a 3-14 membered heterocyclic group means a heterocyclic group consisting of 3-14 ring-forming atoms of C, N, O or S; more preferably a 5-14 membered heterocyclic group and a 3-8 membered heterocyclic group, still more preferably a 3-6 membered heterocyclic group. Wherein the nitrogen or sulfur heteroatoms may be selectively oxidized and the nitrogen heteroatoms may be selectively quaternized. Examples of such heterocyclic groups include, but are not limited to Azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, oxopiperidinyl, tetrahydrofuranyl, dioxolanyl, tetrahydroimidazolylGroup, tetrahydrothiazolyl, tetrahydrooxazolyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, and tetrahydrooxadiazolyl. The heterocyclic group may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is a heterocyclic group.

The term "heteroaryl", in the present invention, unless otherwise indicated, refers to a monocyclic or polycyclic (e.g., having 2, 3 or 4 fused rings, spiro rings, bridged rings, etc.) aromatic heterocycle having at least one heteroatom selected from N, O and/or S, and wherein the nitrogen or sulfur heteroatom may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. Preferably a 5-18 membered heteroaryl group, wherein "5-18 membered" in a 5-18 membered heteroaryl group refers to a heteroaryl group consisting of 5-18 ring-forming atoms of C, N, O or S, more preferably a 5-10 membered heteroaryl group, examples of heteroaryl groups include, but are not limited to, thienyl, furyl, imidazolyl, isoxazolyl, oxazolyl, pyrazolyl, pyrrolyl, thiazolyl, thiadiazolyl, triazolyl, pyridyl, pyridazinyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, benzofuranyl, benzothienyl, benzisoxazolyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyladenine, quinolinyl or isoquinolinyl. The heteroaryl group may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is a heteroaryl ring.

The term "cycloalkyl" refers to a ring system having at least one cyclized alkyl group. Preferably C _3-14 Cycloalkyl radicals, where "C _3-14 "means that the cycloalkyl group may have 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms. Cycloalkyl groups may include monocyclic and polycyclic (e.g., having 2, 3, or 4 fused rings, spiro rings, bridged rings, etc.). Some embodiments include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and the like; the cycloalkyl groups may also be fused to aryl, heterocyclyl or heteroaryl rings, wherein the ring attached to the parent structure is cycloalkyl.

The term "fetchSubstituted "means that one or more hydrogen atoms in the group are each replaced by the same or different substituents. Typical substituents include, but are not limited to, halogen (F, cl, br or I), C _1-8 Alkyl, C _3-12 Cycloalkyl, -OR ¹ 、-SR ¹ 、＝O、＝S、-C(O)R ₁ 、-C(S)R ¹ 、＝NR ¹ 、-C(O)OR ¹ 、-C(S)OR ¹ 、-NR ¹ R ² 、-C(O)NR ¹ R ² Cyano, nitro, -S (O) ₂ R ¹ 、-O-S(O ₂ )OR ¹ 、-O-S(O) ₂ R ¹ 、-OP(O)(OR ¹ )(OR ² ) The method comprises the steps of carrying out a first treatment on the surface of the Wherein R is ¹ And R is ² Independently selected from-H, C _1-6 Alkyl, C _1-6 Haloalkyl or C _3-6 Cycloalkyl groups. In some embodiments of the present invention, in some embodiments, the substituents are independently selected from the group consisting of-F, -Cl, -Br, -I, -OH, trifluoromethoxy, ethoxy propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, -SCH ₃ 、-SC ₂ H ₅ Formaldehyde, -C (OCH) ₃ ) Cyano, nitro, -CF ₃ 、-OCF ₃ Amino, dimethylamino, methylthio, sulfonyl and acetyl groups.

When the number of one linking group is 0, such as- (CH) ₂ ) ₀ -representing that the linking group is a bond.

The term "pharmaceutically acceptable salt" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids.

When the compounds provided herein are acids, the corresponding salts thereof can be conveniently prepared from pharmaceutically acceptable non-toxic bases, including inorganic and organic bases. Salts derived from inorganic bases include salts of aluminum, ammonium, calcium, copper (both higher and lower), ferric, ferrous, lithium, magnesium, manganese (both higher and lower), potassium, sodium, zinc and the like. Particularly preferred are salts of ammonium, calcium, magnesium, potassium and sodium. Nontoxic organic bases capable of derivatizing into pharmaceutically acceptable salts include primary, secondary and tertiary amines, as well as cyclic amines and substituent-containing amines, such as naturally occurring and synthetic substituent-containing amines. Other pharmaceutically acceptable non-toxic organic bases capable of salt formation include ion exchange resins as well as arginine, betaine, caffeine, choline, N' -dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, reduced glucosamine, histidine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, chloroprocaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.

When the compounds provided by the present invention are bases, the corresponding salts thereof can be conveniently prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, formic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, oxalic acid, propionic acid, glycolic acid, hydroiodic acid, perchloric acid, cyclamic acid, salicylic acid, 2-naphthalenesulfonic acid, saccharin acid, trifluoroacetic acid, tartaric acid, p-toluenesulfonic acid, and the like. Preferably, citric acid, hydrobromic acid, formic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid and tartaric acid. More preferably formic acid and hydrochloric acid.

Prodrugs of the compounds of the present invention are included within the scope of the present invention. Typically, the prodrug refers to a functional derivative that is readily converted in vivo to the desired compound. For example, any pharmaceutically acceptable salt, ester, salt of an ester, or other derivative of a compound of the invention, which upon administration to a subject is capable of providing, directly or indirectly, a compound of the invention or a pharmaceutically active metabolite or residue thereof.

The compounds of the present invention may contain one or more asymmetric centers and may thus produce diastereomers and optical isomers. The present invention includes all possible diastereomers and racemic mixtures thereof, substantially pure resolved enantiomers thereof, all possible geometric isomers, and pharmaceutically acceptable salts thereof.

Where tautomers exist for the compounds of the invention, the invention includes any of the possible tautomers and pharmaceutically acceptable salts thereof, and mixtures thereof, unless specifically stated otherwise.

Drawings

FIG. 1 is a graph showing comparison of tumor growth inhibition by compound 7 of example 2 administered in combination with Elacridar.

Wherein, G1 is a Vehicle control group; g2 is compound 7 alone; g3 is compound 7 in combination with Elacridar (day 17-33).

FIG. 2 is a plot of the bulk polarized light microscopy of a single crystal sample of compound M23-1.

FIG. 3 is a single crystal structure analysis chart of the compound M23-1.

Detailed Description

In order to make the above matters clearer and more obvious, the following examples are provided to further illustrate the technical aspects of the present invention. The following examples are presented only to illustrate specific embodiments of the invention so that those skilled in the art can understand the invention and are not intended to limit the scope of the invention. In the specific embodiment of the present invention, technical means, methods, and the like not specifically described are conventional technical means, methods, and the like in the art.

All temperatures in this invention are in degrees celsius unless otherwise indicated.

The following abbreviations are used in the examples:

(BOC) ₂ o: di-tert-butyl dicarbonate;

[PdCl ₂ (dppf)]CH ₂ Cl ₂ : [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane complex;

[Rh(COD)Cl] ₂ : (1, 5-cyclooctadiene) rhodium (I) chloride dimer;

B ₂ Pin ₂ : pinacol ester of biboron acid;

BF ₃ ·Et ₂ o: boron trifluoride diethyl etherate;

BH ₃ THF: borane-tetrahydrofuran solution;

BINAP:1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine;

BOP: a catter condensing agent;

CataCXium A Pd G3: methanesulfonic acid [ n-butylbis (1-adamantyl) phosphine ] (2-amino-1, 1' -biphenyl-2-yl) palladium (II);

CBZCl: benzyl chloroformate;

CDI: n, N' -carbonyldiimidazole;

CH ₃ CN: acetonitrile

CPME: cyclopentyl methyl ether;

Cs ₂ CO ₃ : cesium carbonate;

CsF: cesium fluoride;

DABCO: triethylene diamine;

DBU:1, 8-diazabicyclo undec-7-ene;

DCM: dichloromethane;

dioxane: a dioxane;

DIPEA: n, N-diisopropylethylamine;

DMAP: 4-dimethylaminopyridine;

DME: ethylene glycol dimethyl ether;

DMF: n, N-dimethylformamide;

DMSO: dimethyl sulfoxide;

DPPA: diphenyl azide phosphate;

EA: ethyl acetate;

ESI-MS: electrospray ionization mass spectrometry;

LAH: lithium aluminum hydride;

LiHMDS: lithium bis (trimethylsilyl) amide;

m-CPBA: m-chloroperoxybenzoic acid;

MeOH: methanol;

MTBE: methyl tertiary butyl ether;

NaH: sodium hydride;

n-BuLi: n-butyllithium;

NCS: n-chlorosuccinimide;

NIS: n-iodosuccinimide;

NMP: n-methylpyrrolidone;

Pd(DPEPhos)Cl ₂ : bis (diphenylphosphinophenyl ether) palladium (II) dichloride;

Pd(dtbpf)Cl ₂ : dichloro [1,1' -bis (ear tert-butylphosphine) ferrocene palladium (II);

Pd(PPh ₃ ) ₄ : tetraphenylphosphine palladium;

PdCl ₂ (dppf): [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride;

PE: petroleum ether;

POCl ₃ : phosphorus oxychloride;

Pre-TLC: thin layer chromatography;

PyBOP: 1H-benzotriazol-1-yloxy tripyrrolidinyl hexafluorophosphate;

SOCl ₂ : thionyl chloride;

TBDPSCl: t-butyldiphenylchlorosilane;

TBSCl: t-butyldimethylchlorosilane;

t-BuOH: t-butanol;

t-BuOK: potassium tert-butoxide;

TEA: triethylamine;

Tf ₂ o: trifluoro methanesulfonic anhydride;

TFA: trifluoroacetic acid;

THF: tetrahydrofuran;

TMP:2, 6-tetramethylpiperidine;

and (3) Tol: toluene;

TosMIC: p-methylsulfonylmethyl isonitrile;

synthesis of intermediate compound M1:

step 1: synthesis of Compound M1-1

Dissolving compound M1-0 (208 g) in anhydrous MeOH (2L) at room temperature, dropwise adding thionyl chloride (286 mL) at 0deg.C, reacting at 5deg.C for 1 hr, concentrating the reaction solution after the reaction is completed, adding anhydrous DCM (1L) for dilution, dropwise adding the diluted solution into saturated sodium bicarbonate solution at 0deg.C, separating, washing the organic layer with saturated saline (500 mL), and drying Drying over sodium sulfate, filtering, and concentrating. The concentrate was purified by column chromatography (EA: dcm=0-50%) to give compound M1-1 (240 g,95% yield). ESI-MS m/z=258.1 [ m+h ]] ⁺ 。

Step 2: synthesis of Compound M1-2

Compound M1-1 (235 g) was dissolved in anhydrous THF (2.4L) at room temperature, lithium aluminum hydride (69.4 g) was added in portions at 0℃and stirred at 60℃for 30min after the addition. After the reaction was completed, the reaction solution was cooled, water (69.4 mL) was added dropwise under ice bath, then 15% aqueous sodium hydroxide solution (69.4 mL) was added dropwise, finally water (208.2 mL) was added dropwise, dried over anhydrous sodium sulfate, and the filtrate was filtered and concentrated to give Compound M1-2 (165 g,90% yield) which was used directly in the next step. ESI-MS m/z=202.1 [ m+h ]] ⁺ 。

Step 3: synthesis of Compound M1-3

Compound M1-2 (160 g) was dissolved in trifluoroacetic acid (500 mL), water (67 mL) was added, the reaction was carried out overnight at 60℃and the reaction liquid was concentrated to give crude compound M1-3 (320 g, 259%) which was used directly in the next step. ESI-MS m/z=156.1 [ m+h ]] ⁺ 。

Step 4: synthesis of Compounds M1-4

Compound M1-3 (308 g) was dissolved in DMF (350 mL) at room temperature, imidazole (540 g) was added at 0deg.C, TBDPSCl (170 mL) was added dropwise, and the mixture was stirred at room temperature for 1 hour after the addition was completed. After the reaction was completed, water and EA were added to dilute, and the aqueous phase was extracted with EA 3 times. The organic phases were combined, washed three times with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. Purification of the concentrate by column chromatography (EA: pe=0-15%) gave compound M1-4 (192 g,25% yield). ESI-MS m/z=394.1 [ M+H ] ] ⁺ 。

Step 5: synthesis of Compound M1

Dissolving compound M1-4 (187 g) and difluoromethyl (2-pyridyl) sulfone (184 g) in anhydrous DMF (1.4L) at room temperature, dropwise adding a solution of potassium tert-butoxide (107 g) in DMF (460 mL) at-50deg.C, controlling the temperature at-40deg.C for 2 hours after the dropwise addition, dropwise adding saturated ammonium chloride solution at-50deg.C until the solution becomes weak acid, naturally heating to room temperature for 18 hours, filtering to obtain filtrate, adding EA (1.4L) for dilution, and filtering to obtain filtrateConcentrating. The concentrate was purified by column chromatography (MeOH: dcm=0-10%) to give compound M1 (60 g,67% yield). ¹ H NMR(500MHz,DMSO-d6)δ3.95-3.92(m,1H),3.70-3.67(m,1H),3.32-3.27(m,2H),2.94-2.89(m,1H),2.69-2.66(m,1H),2.50-3.45(m,1H),1.99-1.92(m,2H),1.88-1.75(m,2H)。ESI-MS m/z＝190.1[M+H] ⁺ 。

Synthesis of intermediate compound M2:

step 1: synthesis of Compound M2-1

2-chloro-3-fluoro-pyridine-4-carboxylic acid (54.00 g), toluene (390.00 mL), t-butanol (390.00 mL), triethylamine (128.27 mL), powdered 4A molecular sieves (90 g) (preactivation), and reflux under nitrogen at an internal temperature of 87 ℃ C.) were added sequentially at room temperature. Then cooled to room temperature naturally, DPPA (99.44 mL) was added, the mixture was warmed to reflux, and the reaction was continued for 5 hours. The reaction mixture was cooled to below 40 ℃ and then diluted with 500mL EA; cooling to room temperature, filtering with diatomite, and filtering to remove the added molecular sieve; and using EA1500mL to rinse filter residues for many times and pump out; collecting filtrate, sequentially washing with 700mL of water and 700mL of saturated saline solution, and separating; the organic phase was dried over anhydrous sodium sulfate; filtration, desiccant removal, concentration, column chromatography separation and purification of the concentrate (PE/ea=30:1-20:1), concentration of the eluate, and final product of compound M2-1 (68.2 g, 89.88%). ESI-MS m/z 247.1[ M+H ] ] ⁺ 。

Step 2: synthesis of Compound M2-2

Compound M2-1 (65.00 g) was dissolved in CH at room temperature ₃ CN (82.00 mL) was cooled in a water bath, hydrochloric acid/dioxane (4M/L) (264 mL) was slowly added, and the mixture was stirred at room temperature to react for about 16 hours, whereby a white solid was precipitated and suspended. The reaction mixture was filtered and the filter cake was rinsed with a small amount of acetonitrile, drained and the filtrate was discarded. Collecting a filter cake, adding the filter cake into a mixture of 700mL of saturated sodium bicarbonate aqueous solution and 700mL of ethyl acetate, alkalizing, extracting and separating liquid; extracting the water phase with 350mL of ethyl acetate, and separating; combining acetic acidAdding 300mL of saturated sodium chloride aqueous solution into the ethyl ester phase for washing and separating; the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give compound M2-2 (36.3 g, 94.0% yield). ESI-MS m/z 147.1[ M+H ]] ⁺ 。

Step 3: synthesis of Compound M2-3

Compound M2-2 (36.00 g) was dissolved in acetonitrile (180.00 mL) at room temperature, NIS (66.32 g) and p-toluenesulfonic acid (2.12 g) were added thereto, and the mixture was heated and incubated at 70℃under nitrogen protection. Cooling the reaction solution to 50 ℃, adding 900mL of water, precipitating pink solid powder, and pulping for half an hour; filtering, rinsing the filter cake with water, and pumping. The filter cake was collected, added with 1200mL of ethyl acetate and dissolved completely, then washed twice with 350mL of saturated aqueous sodium sulfite, washed twice with 350mL of saturated aqueous sodium chloride, separated, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give Compound M2-3 (63.2 g, yield 94.43%). ESI-MS m/z 272.9[ M+H ] ] ⁺ 。

Step 4: synthesis of Compound M2-4

Compound M2-3 (57.50 g) was dissolved in DMF (22.00 mL) at room temperature, zinc cyanide (32.22 g), palladium tetraphenylphosphine (12.19 g) and powderedMolecular sieves (20.00 g) were added thereto and reacted at 100℃for about 7 hours under nitrogen atmosphere with heating. And removing the oil bath, naturally cooling to room temperature, and waiting for post-treatment. Filtering with diatomite, filtering the reaction mixture, and pumping; collecting filtrate, concentrating at 60-70 ℃ to obtain a light yellow solid crude product. Rinsing the filter residue with 500mL of ethyl acetate, and draining; collecting the rinsing liquid, combining the rinsing liquid with the crude product, and concentrating again until no liquid is distilled off; 700mL of ethyl acetate was added, dissolved and concentrated to obtain a crude solid, which was then washed 3 times with 250mL of saturated sodium chloride each time, and separated. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated to give a pale yellow solid, 160mL of PE/ea=3/1 mixture was added, slurried for half an hour, filtered and drained. Collecting filter cake, concentrating in 45 deg.C water bath, and pumping with high vacuum oil pump to constant weight; compound M2-4 (36.1 g, 99.7% yield) was finally obtained. ESI-MS m/z 172.0[M+H] ⁺ 。

Step 5: synthesis of Compound M2-5

In a 500mL single-necked flask at room temperature, concentrated sulfuric acid (61.37 mL) was added, the flask was cooled to 10℃or lower by an ice-water bath, and Compound M2-4 (39.30 g) was added in portions, followed by stirring for 10 minutes, and the flask was incubated with an oil bath at 60℃under a nitrogen atmosphere to react for about 1 hour. The reaction was cooled to room temperature and then carefully added to 1100mL of an ice-water mixture, diluted and quenched with a small amount of yellow solid precipitated. Stirring for 10 min, and filtering; collecting filter cake, pulping with 50mL saturated sodium bicarbonate water solution for 20 min, filtering again, collecting the two filtrates, and mixing; then slowly adding sodium carbonate solid, regulating pH to about 7, and separating out off white solid powder. Stirring for half an hour, filtering and pumping; the filter cake was rinsed with 100mL of water each time, drained, and rinsed 2 times in total. The cake was collected and placed in a vacuum oven and dried to constant weight at 55℃to give Compound M2-5 (33.6 g, 77.37% yield). ESI-MS m/z 190.0[ M+H ] ] ⁺ 。

Step 6: synthesis of Compound M2-6

Tetrahydrofuran (470.00 mL) is added at room temperature, after nitrogen replacement, sodium hydride (10.00 g) is added under the protection of micro nitrogen flow, and the mixture is heated by an oil bath, kept at 40-45 ℃ and stirred for 15 minutes; then, compound M2-5 (18.95 g) was added in portions, and after the addition was completed, the mixture was stirred mechanically at an elevated temperature for 20 minutes, CDI (24.31 g) was then added in portions carefully, and after the addition was completed, the mixture was stirred for 15 minutes, the mixture was heated in an oil bath and was refluxed at an elevated temperature. The reaction solution is cooled to below 10 ℃ by ice water bath, then 500mL of saturated ammonium chloride aqueous solution is added, pale yellow solid is separated out, and 1000mL of water is added; then transferring to a 5L beaker, and adding 3000mL of water; stirring for 1 hour, filtering, and pumping; the filter cake was collected and placed in a vacuum oven and dried to constant weight at 50-55℃to give Compound M2-6 (18.3 g, yield 84.93%). ESI-MS m/z 216.0[ M+H ]] ⁺ 。

Step 7: synthesis of Compound M2-7

Compound M2-6 (18.00 g) and DIEA (36.00 mL) were dissolved in POCl at room temperature ₃ (180.00 mL) was heated and incubated at 100deg.C for about 2.5 hours under nitrogen. Concentrating under reduced pressure to remove threePhosphorus oxychloride and taken 2 times with 100mL of DCM; the concentrated residue was dissolved with 400mL of dichloromethane and then added dropwise to 500mL of saturated aqueous sodium bicarbonate solution, cooled with ice water; after stirring for 15 minutes, separating the liquid; extracting the water phase with 300mL of dichloromethane, and separating; the methylene chloride phases are combined, washed and separated by 300mL of saturated sodium chloride aqueous solution; drying over anhydrous sodium sulfate, filtering, concentrating, and purifying the concentrate by silica gel column (PE/ea=90/10 to 75/25) to obtain compound M2-7 (10.95 g, yield 51.94%). ESI-MS m/z 251.9[ M+H ] ] ⁺ 。

Step 8: synthesis of Compound M2

Compound M2-7 (10.50 g) and DIEA (17.18 mL) were dissolved in DCM (120.00 mL) at room temperature, cooled in a water bath, tert-butyl 3, 8-diazabicyclo [3.2.1] octane-8-carboxylate (9.27 g) was added in portions, and the reaction was stirred at room temperature for about 10 minutes. 120mL of methylene chloride was added, and the organic phase was washed with 100mL of water, separated with 100mL of saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography on silica gel (PE/EA=90/10 to 75/25) and slurried (40 mL EA+160mL PE) to give compound M2 (15.9 g, 89.26% yield).

Synthesis of intermediate compound M3:

step 1: synthesis of Compound M3-1

Compound M3-0 (500 mg), styrene (373 mg) and Grubbs generation 2 catalyst (405.72 mg) were dissolved in anhydrous DCM (20 mL) at room temperature and reacted overnight at 50 ℃. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the concentrate was separated and purified by column chromatography to give the objective compound M3-1 (400 mg).

Step 2: synthesis of Compound M3

Compound M3-1 (400 mg) was dissolved in anhydrous THF (10 mL) at room temperature, LAH (160 mg) was added at 0deg.C, and the mixture was allowed to react at 70℃for 30min. After the reaction was completed, the temperature was lowered to 0 ℃, 170uL of water was slowly added, 170uL of 15% aqueous sodium hydroxide solution was then added, 510uL of water was finally added, the reaction was carried out at room temperature for 15min, anhydrous sodium sulfate was added for drying, celite filtration, and the filtrate was concentrated to obtain the objective compound M3 (277 mg).

Synthesis of intermediate compound M4:

step 1: synthesis of Compound M4-1

Compound M3-0 (800 mg), p-chlorostyrene (795 mg) and Grubbs generation 2 catalyst (325 mg) were dissolved in anhydrous DCM (20 mL) at room temperature and reacted overnight at 50 ℃. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the concentrate was separated and purified by column chromatography to give the objective compound M4-1 (729 mg). ESI-MS m/z=320.1 [ m+h ]] ⁺ 。

Step 2: synthesis of Compound M4

Compound M4-1 (729 mg) was dissolved in anhydrous THF (10 mL) at room temperature, LAH (260 mg) was added at 0deg.C, and the mixture was allowed to react at 70℃for 30min. After the reaction was completed, the temperature was lowered to 0 ℃, 280uL of water was slowly added, 280uL of 15% aqueous sodium hydroxide solution was then added, 840uL of water was finally added, the reaction was carried out at room temperature for 15 minutes, anhydrous sodium sulfate was added for drying, celite filtration, and the filtrate was concentrated to obtain the objective compound M4 (563 mg). ESI-MS m/z=264.1 [ m+h ]] ⁺ 。

Synthesis of intermediate compound M5:

step 1: synthesis of Compound M5-1

Compound M3-0 (400 mg), methylenecyclopentane (236 mg) and Grubbs generation 2 catalyst (325 mg) were dissolved in anhydrous DCM (20 mL) at room temperature and reacted overnight at 50 ℃. After the reaction, the reaction mixture was concentrated to a small amount under reduced pressure, and the mixture was directly subjected to wet-process chromatography to obtain the objective compound M5-1 (410 mg).

Step 2: synthesis of Compound M5

Compound M5-1 (410 mg) was dissolved in anhydrous THF (10 mL), LAH (204 mg) was added at 0deg.C, and the mixture was reacted at 70℃for 30min. After the reaction was completed, the temperature was lowered to 0 ℃, 210uL of water was slowly added, then 210uL of 15% aqueous sodium hydroxide solution was added, 630uL of water was added, the reaction was carried out at room temperature for 15 minutes, anhydrous sodium sulfate was added for drying, celite filtration, and the filtrate was concentrated to obtain the objective compound M5 (370 mg).

Synthesis of intermediate compound M6:

step 1: synthesis of Compound M6-1

M3-0 (1.00 g), 2, 3-dimethyl-2-ene (2.01 g) was added to DCM (20.00 mL) at room temperature, followed by Grubbs generation 2 catalyst (0.41 g). N (N) ₂ Three times of replacement, N ₂ The reaction was stirred for 20 hours at 50℃under protection. The reaction is cooled and concentrated under reduced pressure, and the concentrate is purified by column chromatography to obtain the target compound M6-1.

Step 2: synthesis of Compound M6

M6-1 (0.20 g) was added to THF (4.00 mL) at room temperature, and LAH (0.10 g) was added in portions under an ice bath. The reaction was stirred for 1 hour at 70 ℃. Cooling the reaction, and dropwise adding H under ice bath ₂ O (100 uL), followed by dropwise addition of 15% NaOH (100 uL) solution, and finally dropwise addition of H ₂ O (300 uL), stirred at room temperature for 10 minutes, dried over anhydrous sodium sulfate and stirred for 5 minutes. The mixture was filtered, and the cake was washed with EA, and the filtrate was concentrated under reduced pressure to give the objective compound M6 (0.18 g). Synthesis of intermediate compound M7:

Step 1: synthesis of Compound M7-1

Compound M3-0 (0.5 g), togni's reagent II (1.51 g) and tetrabutylammonium iodide (0.44 g) were dissolved in 1, 4-dioxane (10 mL) at room temperature, nitrogen was purged, the reaction solution was directly concentrated at 80℃for 10 hours, and the concentrate was separated and purified by a column chromatography (DCM/EA=2/1) to give compound M7-1 (0.45 g,68% yield).

Step 2: synthesis of Compound M7

Compound M7-1 (0.3 g) was dissolved in tetrahydrofuran at room temperature, lithium aluminum hydride (123 mg) was added under ice bath, then a solution of borane in tetrahydrofuran (3.2 mL) was added, the reaction was immediately quenched with water after 1 minute until no more bubbles were evolved, 15% sodium hydroxide (0.3 mL) was added, water (0.9 mL) was further added, after completion of stirring quenching, sodium sulfate was added to dry, the filtrate was filtered, and concentrated to give compound M7 (80 mg,33% yield).

Synthesis of intermediate compound M8:

compound M7-1 (0.3 g) was dissolved in tetrahydrofuran at room temperature, lithium aluminum hydride (123 mg) was added under ice bath, then a solution of borane in tetrahydrofuran (3.2 mL) was added, the ice bath reaction was removed for 5 minutes, LC-MS monitored for completion of the reaction, the reaction was quenched with water until no more bubbles emerged, 15% sodium hydroxide (0.3 mL) was added, water (0.9 mL) was further added, after completion of stirring and quenching, sodium sulfate was added to dry, and the filtrate was filtered and dried by spinning to give compound M8 (164 mg,73% yield).

Synthesis of intermediate compound M9:

ethyl 2-methylene-5-oxo-1, 3,6, 7-tetrahydropyrrolizine-8-carboxylate (10.00 g) was dissolved in THF (150.00 mL) at room temperature in a reaction flask, LAH (3.63 g) was slowly added and the temperature was controlled below 60℃and stirring was completed for 0.2h. Cooling to 0 ℃, adding 3.6ml of water to quench the reaction, adding 3.6ml of 15% sodium hydroxide aqueous solution, finally adding 10.8ml of water, stirring for 10min, adding anhydrous magnesium sulfate to dry, stirring for 10min, filtering, washing a filter cake with EA for three times, and concentrating a mother liquor to obtain a target intermediate compound M9 (6.5 g,89.34% yield).

Synthesis of intermediate compound M10:

step 1: synthesis of Compound M10-1

1-bromo-2, 5-difluoro-3-nitrobenzene was dissolved in ethanol (400 ml) and water (80 ml), ammonium chloride (28.1 g) was added, iron powder (20.53 g) was added with stirring, and the mixture was reacted at 75℃for 2 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, filtered through celite, the residue was washed with DCM, the filtrate was washed once with water, saturated brine was washed once, the organic phase was dried, filtered and concentrated to give compound M10-1 (21.13 g yield 96.70%). The crude product was used directly in the next step. ESI-MS m/z 208.13[ M+H ]] ⁺ 。

Step 2: synthesis of Compound M10-2

Hydroxylamine hydrochloride (24.71 g) and chloral hydrate (25.2 g) were dissolved in water (435 ml), and anhydrous sodium sulfate (115.43 g) was added with stirring. M10-1 (21.13 g) was dissolved in ethanol (61 ml) and water (35 ml), concentrated hydrochloric acid (14.7 ml) was added, and the mixture was added to a reaction flask and reacted overnight at 60 ℃. After completion of the reaction, the reaction mixture was filtered under heating to give a cake, which was dried to give Compound M10-2 (25.10 g yield 88.55%). The crude product was used directly in the next step.

Step 3: synthesis of Compound M10-3

Compound M10-2 (25.10 g) was added to concentrated sulfuric acid (250 ml) at 60℃and the temperature was raised to 90℃after the addition. The reaction was carried out for 1h. After the reaction was completed, the reaction solution was cooled, slowly added to ice water, a large amount of solids was precipitated, and the solid was obtained by filtration, dissolved with EA, washed twice with saturated brine, dried, filtered, and spin-dried to give Compound M10-3 (16.07 g yield 68.19%). The crude product was used directly in the next step.

Step 4: synthesis of Compound M10-4

Sodium hydroxide (22.08 g) was dissolved in water (280 ml), added to a reaction flask containing compound M10-3 (16.07 g), the temperature was lowered to 0℃after the addition, hydrogen peroxide (31 ml) was added, and the reaction was allowed to proceed to room temperature overnight. After the reaction is completed, the pH is adjusted to 7 by using concentrated hydrochloric acid, the mixture is filtered to obtain filtrate, The mixture was further adjusted to pH 1 with concentrated hydrochloric acid, filtered to give a solid, dissolved with EA, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give Compound M10-4 (12.54 g yield 81.13%). ESI-MS m/z 252.14[ M+H ]] ⁺ 。

Step 5: synthesis of Compound M10-5

Compound M10-4 (12.34 g) was dissolved in DMF (120 ml), N-chlorosuccinimide (7.85 g) was added and reacted at 70℃for 0.5h. After completion of the reaction, the reaction mixture was cooled to 0℃and water (480 ml) was slowly added dropwise to give a large amount of solid, which was filtered to give a solid, which was dissolved in EA, washed with saturated brine once, dried, filtered, and concentrated to give Compound M10-5 (12.95 g, yield 92.32%). ESI-MS m/z 286.1[ M+H ]] ⁺ 。

Step 6: synthesis of Compound M10-6

Compound M10-5 (12.46 g) was dissolved in THF (120 ml), CDI (10.58 g) was added, and the reaction was carried out at 50℃for 0.5h, the reaction solution was cooled to room temperature, and slowly added dropwise to aqueous ammonia (120 ml) of ice, and the reaction was continued at room temperature for 0.5h. After completion of the reaction, the reaction mixture was diluted with EA and water, extracted with EA, saturated brine was concentrated by drying, and the concentrate was purified by column chromatography to give Compound M10-6 (9.64 g, yield 77.63%). ESI-MS m/z 285.12[ M+H ]] ⁺ . Step 7: synthesis of Compound M10-7

Compound M10-6 (4 g) was dissolved in THF (50 ml), the temperature was raised to 40℃and NaH (1.4 g) was added in portions, stirred at 40℃for 10 minutes, N' -thiocarbonyldiimidazole (3.75 g) was added in portions, and the reaction was completed by raising the temperature to 60℃for 0.5h. After the reaction was completed, the reaction mixture was quenched with saturated ammonium chloride, the pH was adjusted to 5-6 with dilute hydrochloric acid, tetrahydrofuran was removed by vacuum concentration, a large amount of solids were precipitated, and the solid was obtained by filtration and dried to give Compound M10-7 (3.79 g yield 82.58%).

Step 8: synthesis of Compound M10

Compound M10-7 (3.59 g) was dissolved in methanol (60 ml), and sodium methoxide (0.89 g) and methyl iodide (1.36 ml) were added thereto to react at room temperature for 0.5h. After completion of the reaction, water (10 ml) was added to the reaction mixture, stirred for 10 minutes, and the solid was obtained by filtration, followed by drying to obtain Compound M10 (2.79 g yield 74).53％)。ESI-MS m/z:341.09[M+H] ⁺ 。

Synthesis of intermediate compound M11:

step 1: synthesis of Compound M11-1

4-bromo-2-fluoroaniline (11.0 g), potassium carbonate (20.18 g) and KI (9.7 g) were added to NMP (110.0 mL) respectively, and 1- (chloromethyl) -4-methoxybenzene (16.24 mL) was added dropwise thereto and stirred at room temperature for 7 hours. Pouring the reaction solution into H ₂ O (120 mL) was extracted 2 times with MTBE (60 mL) (three layers were separated, the upper layer was taken), and the organic layer was washed twice with saturated saline and dried over anhydrous sodium sulfate. The crude product was slurried with PE (60 ml) for 1 hour. Filtering, washing the filter cake with PE, and drying to obtain the compound M11-1 (17.68 g,70.33% yield). ESI-MS m/z=430 [ M+H ] ] ⁺ 。

Step 2: synthesis of Compound M11-2

The above compound M11-1 (6.3 g) was added to the reaction flask, followed by pinacol diboronate (11.2 g), potassium acetate (2.88 g) and Pd (dppf) Cl ₂ DCM (1.20 g), 100℃for 12h. TLC confirmed complete conversion of starting material. Ethyl acetate (30 mL) was added thereto, followed by washing with a saturated aqueous sodium chloride solution (20 mL. Times.2) and liquid separation. Anhydrous sodium sulfate was added to dry. Filtration, concentration, and separation and purification of the concentrate by column chromatography (pe—pe: ea=10:1) gave compound M11-2 (4.14 g,67.5% yield). ESI-MS m/z=478 [ M+H ]] ⁺ 。

Step 3: synthesis of Compound M11-3

The product M11-2 (4.1 g) of the above step was dissolved in dioxane (50 mL) and water (10 mL) and 4-methyl-2-en-1-one (3.2 g), BINAP (1.22 g) and [ Rh (COD) Cl were added to the flask] ₂ (483mg)，K ₃ PO ₄ (6.2 g) was reacted at 35℃for 10 minutes after nitrogen substitution. TLC monitoring showed complete reaction of the starting material, 40mL of ethyl acetate was added, followed by 30 mL. Times.2 washing with saturated aqueous sodium chloride solution and separation. Anhydrous sodium sulfate was added to dry. Filtering, concentrating, and separating and purifying the concentrate by column chromatography (PE-PE: EA=10:1) to obtain a compound M11-3(2.84 g,62.8% yield). ESI-MS m/z=462 [ m+h ]] ⁺ 。

Step 4: synthesis of Compound M11-4

The product M11-3 (2.6 g) of the previous step was added to the reaction flask, acetonitrile (25 mL) was added for dissolution, NIS (2.05 g) and trifluoroacetic acid (68 mg) were added for reaction at room temperature for 2 hours, the solvent was concentrated, and the concentrate was separated and purified by column chromatography (PE.about.PE: EA=8:1) to give compound M11-4 (1.22 g,33.7% yield). ESI-MS m/z=588 [ m+h ] ] ⁺ 。

Step 5: synthesis of Compound M11-5

To the reaction flask, M11-4 (1.2 g) was added, and DMF (12 mL) was added for dissolution, followed by addition of methyl 2, 2-difluoro-2-fluorosulfonyl acetate (1.56 g) and cuprous iodide (1.16 g), followed by reaction at 85℃for 2 hours under nitrogen protection. Ethyl acetate (30 mL) was added thereto, followed by washing with 25ml×3 of a saturated aqueous sodium chloride solution and liquid separation. Anhydrous sodium sulfate was added to dry. Filtration, concentration, and separation and purification of the concentrate by column chromatography (pe—pe: ea=3:1) gave compound M11-5 (600 mg,55.6% yield). ESI-MS m/z=530 [ M+H ]] ⁺ 。

Step 6: synthesis of Compound M11-6

To the reaction flask was added compound M11-5 (600 mg), dissolved in THF (10 mL), cooled to-78℃under nitrogen protection, then LiHMDS (2.3 mL) was slowly added, and after the addition was completed, the reaction was allowed to stand for 0.4h, then ethyl cyanide (246 mg) was added at that temperature, and after the addition was completed, the reaction was allowed to proceed for 0.5h again at that temperature. TLC monitoring showed complete reaction of the starting material, quenched by addition of 5mL of aqueous ammonium chloride at low temperature, followed by addition of 20mL of ethyl acetate, and washing with 20mL of saturated aqueous sodium chloride by 2 and pipetting. Anhydrous sodium sulfate was added to dry. Filtration and concentration of the filtrate gave crude M11-6 (600 mg 88.3% yield). ESI-MS m/z=602 [ M+H ]] ⁺ 。

Step 7: synthesis of Compound M11-7

The above compound M11-6 (600 mg) was added to ethanol (10 mL) and water (2 mL), and sodium hydrogencarbonate (2.0 g) and methyl isothiourea sulfate (2.2 g) were added, and the reaction solution was stirred at 50℃for 4 hours. The reaction mixture was added to 25mL of water, extracted 2 times with ethyl acetate (15 mL), and the combined organic phases were washed 2 times with saturated brine (15 mL) and dried over anhydrous sodium sulfateFiltering, and concentrating the filtrate under reduced pressure. The concentrate was purified by silica gel column chromatography (PE: ea=100:0-1:1) to give compound M11-7 (480 mg). ESI-MS m/z=628 [ m+h ]] ⁺ 。

Step 8: synthesis of Compound M11

Compound M11-7 (480 mg) was dissolved in methylene chloride (5 mL), N diisopropylethylamine (0.3 g) was added thereto, the temperature was lowered to 0-10℃and trifluoromethanesulfonic anhydride (0.3 g) was slowly added to the reaction mixture, followed by reaction at this temperature for 15 minutes. The reaction solution was poured into saturated aqueous ammonium chloride (5 mL), the solution was separated, the aqueous phase was extracted 2 times with methylene chloride (5 mL), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Pulping the concentrate by a mixed solvent (PE: MTBE=20:1, 4 mL), filtering, and drying a filter cake to obtain the compound M11.ESI-MS m/z=760 [ M+H ]] ⁺ 。

Synthesis of intermediate compound M12:

Step 1: synthesis of Compound M12-1

To a mixed solution of tert-butyl 2, 5-dihydro-1H-pyrrole-1-carboxylate (20 g) in tert-butanol (200 mL) and water (200 mL) at room temperature was added potassium osmium dihydrate (1.74 g) and N-methylmorpholine oxide (51.5 g) in this order, and the mixture was stirred at 45℃for 15 hours. The reaction solution was concentrated directly, extracted 2 times with 50mL each, and washed with a saturated sodium sulfite solution (60 mL). The combined organic layers were washed 3 times with 50mL portions each with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was separated and purified by column chromatography (petroleum ether/ethyl acetate=1/0 to 0/1) to give compound M12-1 (19.0 g). ¹ H NMR(400MHz，CDCl ₃ )δ4.20(m，2H)，3.57-3.54(m，2H)，3.32-3.29(m，2H)，2.85-2.80(m，2H)，1.40(s，9H)。

Step 2: synthesis of Compound M12-2

Compound M12-1 (19 g) was dissolved in methylene chloride (150 mL) at room temperature, cooled to 0℃and iodobenzene diacetate (20.0 g) was then added, and the reaction system was transferred to room temperature and stirred for 3 hours. The reaction was quenched by addition of saturated sodium bicarbonate solution (40 mL), and dichloromethane (10 mL) was added and stirred for 0.5 h, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. Methyl tert-butyl ether (20 mL) was added at 25℃and stirred for 10 min, filtered and concentrated under reduced pressure to give compound M12-2 (19.5 g).

Step 3: synthesis of Compound M12-3

Compound M12-2 (19.5 g) was dissolved in tetrahydrofuran (50 mL) at room temperature, cooled to-78℃and ethyl magnesium bromide (1M, 150 mL) was added to the reaction system. The reaction was warmed to room temperature and stirred for 15 hours. The reaction was then cooled to 10℃and quenched by addition of saturated ammonium chloride solution (100 mL) thereto, followed by extraction with ethyl acetate. The organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified by column chromatography to give Compound M12-3 (18.9 g).

Step 4: synthesis of Compound M12-4

Compound M12-3 (18.9 g) was dissolved in methylene chloride (200 mL) at room temperature, the reaction system was moved to 0℃and DBU (4.5 g) and 2, 2-trichloroacetonitrile (53 g) were added, and the reaction system was transferred to room temperature and stirred for 16 hours. The residue was extracted twice with ethyl acetate, washed twice with water and saturated brine, respectively, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified by column chromatography (petroleum ether/ethyl acetate=1/0 to 5/1) to give compound M12-4 (17.5 g).

Step 5: synthesis of Compound M12-5

The compound 2-phenylpropane-2-amine (6.5 g) was dissolved in DCE (140 mL) at room temperature, and 1, 5-cyclooctadiene iridium chloride dimer (2.5 g) was added thereto. The reaction system was cooled to 0 ℃, compound M12-4 (17.5 g) was dissolved in DCE (200 mL) and added to the above reaction system, the reaction was allowed to warm to room temperature and stirred for further 15 hours, and the reaction solution was concentrated directly. The concentrate was purified by column chromatography (petroleum ether/ethyl acetate=1/0 to 10/1) to give compound M12-5 (15.1 g). ESI-MS m/z=357 [ M+H ]] ⁺ 。

Step 6: synthesis of Compound M12-6

M12-5 (15.1 g) was dissolved in toluene (380 mL) at room temperature followed by the addition of a second generation Grubbs catalyst (1.2 g). The reaction system was warmed to 125 ℃ and stirred for 16 hours. The reaction solution was directly filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate=1/0 to 10/1) to give compound M12-6 (12.1 g). ESI-MS m/z=329 [ M+H ]] ⁺ 。

Step 7: synthesis of Compound M12-7

Compound M12-6 (12.1 g) was dissolved in methanol (90 mL) at room temperature, followed by the addition of HCl/MeOH (4M, 35 mL). The reaction was stirred for 16 hours at 35 ℃. The pH of the reaction mixture was adjusted to 12, and extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give Compound M12-7 (11.0 g). ESI-MS m/z=229 [ M+H ] ] ⁺ 。

Step 8: synthesis of Compound M12-8

Compound M12-7 (11 g) was dissolved in THF (100 mL) at room temperature, followed by addition of 9-fluorenylmethylchloroformate (9.0 g), sodium carbonate (10.0 g). The mixture was stirred at 0 ℃ for 1 hour. Ethyl acetate extraction was twice and water washing was performed once. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give Compound M12-8 (9.8 g). ESI-MS m/z=451 [ m+h ]] ⁺ 。

Step 9: synthesis of Compound M12-9

Compound M12-8 (9.8 g) was dissolved in TFA (180 mL) at room temperature, heated to 75deg.C and stirred for 16 hours. Water (20.0 mL) was added and the pH was adjusted to 9, and dichloromethane (20.0 mL) was added for extraction, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was slurried with n-heptane (6 mL) at 25℃to give trifluoroacetate salt (7.4 g) of Compound M12-9.

Step 10: synthesis of Compound M12-10

Trifluoroacetate salt of Compound M12-9 (7.4 g) was dissolved in tetrahydrofuran (70 mL) at room temperature, followed by the sequential addition of di-tert-butyl dicarbonate (5.0 g), triethylamine (6.0 g) and stirring at 25 ℃And 1 hour. Ethyl acetate and water were added for extraction. The organic phases were combined, washed with saturated brine (50.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified by column chromatography (petroleum ether/ethyl acetate=1/0 to 3/1) to give intermediate compound M12-10 (6.9 g). ESI-MS m/z=433 [ m+h ] ] ⁺ 。

Step 11: synthesis of Compound M12

Compound M12-10 (6.9 g) was dissolved in ethanol (70.0 mL) at room temperature, followed by dimethylamine (36.6 g). The reaction system was stirred at 25℃for 3 hours. The mixture was concentrated directly under reduced pressure, the concentrate was extracted with ethyl acetate (50.0 mL) and 10% citric acid (200.0 mL), the pH of the aqueous phase was adjusted to 9, filtered and extracted 2 times with ethyl acetate, 40.0mL each time, the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound M12 (1.6 g,96.3% purity, 47.7% yield). ESI-MS m/z=211 [ m+h ]] ⁺ 。 ¹ H NMR(400MHz，CDCl ₃ )δ6.20(m，2H)，4.37(m，2H)，2.88-3.00(m，2H)，2.38(m，2H)，1.49(s，9H)。

Synthesis of intermediate compound M13:

the compound ((2-fluoro-6- (methoxymethoxy) -8- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-1-yl) ethynyl) triisopropylsilane (10 g) was added to a 100mL single-necked flask, and 30mL hydrochloric acid (4M/Dioxane) was added. After reacting at room temperature for half an hour, the reaction solution was neutralized by adding a saturated sodium bicarbonate solution in an ice bath, filtered, the cake was washed twice with water, and after dissolving EA, it was dried over anhydrous sodium sulfate, concentrated, and the concentrate was purified by column chromatography (PE: ea=82:18) to give the objective intermediate compound M13 (9.1 g).

The synthesis step of intermediate M14:

step 1: synthesis of Compound M14-1

t-BuOK (27.93 g) was dissolved in DME (100.00 mL) at room temperature, a solution of TosMIC (24.29 g) in DME (100.00 mL) was added thereto at-78℃and a solution of 6-bromo-2, 3-difluorobenzaldehyde (25.00 g) in DME (100.00 mL) was added thereto, and the mixture was stirred at a constant temperature for 1h. The reaction mixture was concentrated directly, water was added, extracted with DCM, the organic phase was concentrated, and the concentrate was purified by column chromatography (EA: pe=0-10%) to give the title compound M14-1 (11.0 g, 41.91%) as a white solid. ESI-MS m/z 232[ M+H ] ] ⁺ 。

Step 2: synthesis of Compound M14-2

Compound M14-1 (10.00 g) was dissolved in DMF (100.00 mL) at room temperature, t-BuOK (5.32 g) was added thereto under ice bath, and the reaction was stirred for 10min to make the solution dark red. Ethyl isothiocyanamide (6.22 g) was added thereto, and the mixture was stirred at room temperature for 1 hour, and then was stirred at 100℃for 30 minutes. After the reaction, the temperature is reduced to 0 ℃ by using an ice bath, water is slowly added, the mixture is filtered, and a filter cake is washed by water and PE and dried, so that the target compound M14-2 (11.0 g, yield 74.37%) is obtained. ESI-MS m/z 343[ M+H ]] ⁺ 。

Step 3: synthesis of Compound M14-3

Compound M14-2 (11.00 g) was dissolved in DMSO (80.00 mL) at room temperature, and NaOH (12.82 g) was added to H ₂ O (60.00 mL) was stirred overnight at 100deg.C. The reaction mixture was diluted with water and extracted with EA. The organic phases were combined, dried and concentrated to give the objective compound M14-3 (11.0 g). ESI-MS m/z 271[ M+H ]] ⁺ 。

Step 4: synthesis of Compound M14-4

Compound M14-3 (15.00 g), DMAP (0.54 g), (BOC) was added at room temperature ₂ O (13.28 g) was dissolved in THF (200.00 mL) and DMF (30.00 mL), DIEA (13.72 mL) was added dropwise thereto, and the mixture was stirred at room temperature for 3h. Water was added to the reaction, extracted with ethyl acetate, and the organic phase was dried and concentrated. The concentrate was slurried with EA: pe=1:5 and filtered to give the title compound M14-4 (7.3 g, 35.54%) as a yellow solid. ESI-MS m/z 371[ M+H ] ] ⁺ 。

Step 5: synthesis of Compound M14

To compound M14-4 (1.00 g) at room temperature)，B ₂ Pin ₂ (2.05g)，[PdCl ₂ (dppf)]CH ₂ Cl ₂ (0.44 g), acOK (1.32 g) was dissolved in 1, 4-dioxane (20.00 mL), and stirred at 80℃for 3h. Water and ethyl acetate are added into the reaction liquid for extraction, and the organic phase is dried and concentrated. The concentrate was purified by column chromatography (EA: pe=3-5%) to give the title compound M14 (750 mg) as a white solid. ESI-MS m/z 419[ M+H ]] ⁺ 。

Synthesis of intermediate M15:

step 1: synthesis of Compound M15-1

2, 6-Dibromobenzaldehyde (25.0 g) was dissolved in ethanol (300 mL) at 0deg.C, sodium borohydride (3.58 g) was added in portions, and after the addition was completed, the mixture was returned to room temperature and stirred for 30min. After completion of the reaction, water (500 mL) was added thereto, and the extracts were twice extracted with DCM (200 mL), and the organic phases were combined, washed once with saturated brine (100 mL), dried over anhydrous sodium sulfate, and spun-dried to give compound M15-1 (25.0 g, yield 99%). ESI-MS m/z=267 [ m+h ]] ⁺ 。

Step 2: synthesis of Compound M15-2

Compound M15-1 (25 g), triphenylphosphine (32.1 g) was dissolved in DCM (300 mL) at room temperature, cooled to 0deg.C, NBS (18.4 g) was added in portions, incubated after addition and stirred for 30min. After completion of the reaction, the reaction solution was concentrated. Purification of the concentrate by column chromatography (EA: pe=0-50%) gave compound M15-2 (20 g, 64% yield).

Step 3: synthesis of Compound M15-3

Compound M15-2 (20 g), potassium cyanide (11.9 g) was dissolved in a mixed solution of ethanol (100 mL) and water (30 mL) at room temperature, and heated under reflux for 3-5h. After completion of the reaction, the reaction mixture was concentrated, diluted with EA (100 mL), washed with water (30 mL), and washed with saturated sodium bicarbonate solution (30 mL) and saturated brine (30 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated. Purification of the concentrate by column chromatography (EA: pe=10%) gave compound M15-3 (13 g, 77% yield).

Step 4: synthesis of Compound M15-4

NaH (3.0 g,60% purity) was added to DMF (130 mL) under ice bath, and Compound M15-3 (20.0 g) was added, followed by stirring for 10min after completion of the addition. Ethyl isothiocyanamide (10.5 g) was then added dropwise to the reaction mixture, followed by stirring for 10min after the completion of the addition, and the mixture was returned to room temperature and stirred for 20min. CuI (1.39 g) and L-proline (1.68 g) were added to the reaction mixture, and after the addition was completed, the temperature was raised to 65℃and the mixture was stirred for 2 to 5 hours. After completion of the reaction, 0.2M aqueous EDTA (500 mL) and EA (100 mL) were added to the reaction mixture, and the mixture was stirred overnight. The mixture was filtered, and the cake was washed twice with water (200 mL) and dried to give Compound M15-4 (23 g, 97% yield). ESI-MS m/z=323 [ M-H ]] ^- 。

Step 5: synthesis of Compound M15-5

Compound M15-4 (23 g) was dissolved in DMSO (100 mL) at room temperature, sodium hydroxide (6M, 90 mL) was added, and the mixture was stirred for 6h at 110 ℃. After completion of the reaction, the reaction mixture was cooled to room temperature, and slowly poured into water (300 mL). Filtration and cake drying gave compound M15-5 (15.0 g, 84% yield). ESI-MS m/z=251 [ m-H ]] ^- 。

Step 6: synthesis of Compound M15-6

Compound M15-5 (15.0 g), DIEA (14.7 mL), DMAP (0.58 g), di-tert-butanol dicarbonate (14.2 g) were added to THF (200 mL) and DMF (30 mL) under ice-bath, and stirred at room temperature for 1h. After the completion of the reaction, the reaction mixture was diluted with 500mL of EA, washed with 300mL of water, 300mL of saturated brine, dried over anhydrous sodium sulfate, and spun-dried. Purification by column chromatography (EA: pe=0-20%) afforded compound M15-6 (16.0 g, 76.4% yield). ESI-MS m/z=351 [ M-H ]] ^- 。

Step 7: synthesis of Compound M15

Compound M15-6 (16.0 g) was dissolved in THF (250 mL) at room temperature, naH (3.6 g,60% purity) was added in portions, and after addition was stirred at room temperature for 30min. Cooling to-78 ℃, dropwise adding n-butyllithium (36 mL, 2.5M), and preserving the heat for 30min after the addition. Methoxy pinacol borate (35.8 g) was added dropwise and slowly returned to room temperature after the addition was completed. The reaction was added dropwise to 800mL of saturated ammonium chloride solution, the solution was separated, extracted with EA (200 mL) in aqueous phase, and the organic phases were combined. The organic phase was washed once with 100mL of saturated brine, Dried over anhydrous sodium sulfate and spun dry. Purification by column chromatography (EA: pe=1-20%) afforded intermediate compound M15 (11 g, 60% yield). ESI-MS m/z=399.2 [ m-H ]] ^- 。

Synthesis of intermediate M16:

step 1: synthesis of Compound M16-1

Di (CDI (2.70 g) was added to the compound 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid (4.0 g) in THF (20 mL) at room temperature, N-ethyl-N-isopropyl-2-amine (1.4 g) was added thereto, the mixture was allowed to react at 50℃for 2 hours, 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid was substantially completely converted into an intermediate product, the mixture was then added dropwise to ice aqueous ammonia (35 mL) and stirred for 5min until the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, and the organic layer was washed with brine, followed by Na ₂ SO ₄ Dried and concentrated in vacuo and the residue was purified by flash column chromatography on silica gel (petroleum ether/ethyl acetate=70:30) to give the desired target product compound M16-1 (1.6 g) as a brown solid.

Step 2: synthesis of Compound M16-2

The mixture of compound M16-1 and urea was reacted at 200℃with stirring for 3 hours. The mixture was cooled to room temperature, the solid was washed with ethyl acetate, and the solid was dried to give the desired crude solid compound M16-2 (209 mg,78% yield).

Step 3: synthesis of Compound M16-3

Adding DIPEA to POCl of M1-2 at room temperature ₃ Is refluxed at 110℃for 16 hours. The mixture was cooled to room temperature and concentrated in vacuo to remove POCl ₃ The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=100:1 to 50:1) to give the target compound M16-3 as a brown solid.

Step 4: synthesis of Compound M16

M16-3 (3.0 g) was dissolved in 1, 4-dioxane (20 ml) at room temperatureAddition of tert-butyl (1R, 5S) -3, 8-diazabicyclo [3.2.1 to the solution]Octane-8-carboxylate (1.6 g) and DIPEA (2.8 g), and the resulting mixture was stirred at room temperature for 10 minutes. The mixture was concentrated in vacuo and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=75:25 to 65:35) to give the title compound M16 (2.5 g,57% yield) as a yellow solid. ESI-MS m/z 505[ M+H ]] ⁺ . Synthesis of intermediate M17:

step 1: synthesis of Compound M17-1

Intermediate M16-1 (19.00 g) was added to tetrahydrofuran (200.00 mL) at room temperature, and sodium hydride (7.10 g) was added in portions at room temperature. N (N) ₂ Under protection, the mixture was stirred for 0.5 hour at 40 ℃. N, N' -thiocarbonyldiimidazole (18.99 g) was added in portions. Stirred for 0.5 hours at a temperature of 70 ℃. LCMS monitored completion of the reaction. Adding saturated NH ₄ The reaction was quenched with Cl (20 mL), adjusted to ph=5-6 with 1N HCl, extracted 2 times with EA (80 mL), the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give 17.4g of crude compound M17-1 as a yellow powder, which was directly used in the next step.

Step 2: synthesis of Compound M17-2

Compound M17-1 (16.40 g) was added to methanol (300.00 mL) and then sodium methoxide (4.29 g) at room temperature, and methyl iodide (4.95 mL) was added dropwise thereto and stirred at room temperature for 0.5 hours. Adding H ₂ Diluting with O (20 mL), adjusting pH to 5-6 with 2N HCl, concentrating under reduced pressure, and adding H into the crude product ₂ O (20 mL) was stirred for 5 min, filtered, the filter cake was washed 3 times with water and then PE/EA=3/1 (20 mL), and dried to give compound M17-2 (14.6 g) as a yellow powder. ESI-MS m/z 323[ M+H ]] ⁺ 。

Step 3: synthesis of Compound M17-3

Compound M17-2 (14.19 g), tert-butyl (1R, 5S) -3, 8-diazabicyclo [3.2.1, was reacted at room temperature]Octane-8-carboxylate (12.10 g) was added acetonitrile (220.00 mL), followed by DBU (32.76 mL) and BOP (58.19 g). Stirred at room temperature for 2 hours. Adding EA (100 mL) andH ₂ o (100 mL) was extracted 2 times, and the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Purification by column chromatography (PE/EA, 10% EA) afforded Compound M17-3 (16 g) as a yellow powder. ESI-MS m/z 517[ M+H ] ] ⁺ 。

Step 4: synthesis of Compound M17-4

Compound M17-3 (11.00 g), intermediate M4 (15.46 g) was added to CPME (165.00 mL) followed by Pd (DPEPhos) Cl at room temperature ₂ (3.04 g) and K ₃ PO ₄ (13.53g)，KF(4.94g)。N ₂ Three times of replacement, N ₂ Under protection, the temperature was raised to 83℃and stirred overnight. EA (80 mL) and H were added ₂ O (100 mL) was extracted 2 times, the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, and the concentrate was purified by column chromatography (PE/EA, 25% EA) to give Compound M17-4 (5.1 g) as a yellow powder, ESI-MS M/z 729[ M+H ]] ⁺ 。

Step 5: synthesis of Compound M17

Compound M17-4 (4.60 g) was added to DCM (100.00 mL) and M-CPBA (4.3 g) was added at room temperature. Stirred at room temperature for 0.5 hours. LCMS monitored completion of the reaction. Adding saturated NaHCO ₃ The pH was adjusted to 8, the excess m-CPBA was quenched with sodium sulfite solution and concentrated under reduced pressure to remove DCM. Extracting with EA for 2 times, 70ml each time, and saturated NaHCO for the organic layer ₃ The resulting mixture was washed with brine, dried over anhydrous sodium sulfate, and concentrated. The concentrate was purified by column chromatography (PE/EA, 50% EA) to give the desired intermediate compound M17 (4.54 g) as a yellow powder. ESI-MS m/z 761[ M+H ]] ⁺ . Synthesis of intermediate compound M18:

step 1: synthesis of intermediate compound M18-1

The compound 1-methoxycarbonylcyclopropane-1-carboxylic acid (5.00 g) was dissolved in DCM (80.00 mL) at room temperature, DMF (0.27 mL) was added in ice bath, and oxalyl chloride (8.81 mL) was added dropwise thereto and stirred at 35℃for 3h after the addition. And finally, directly concentrating the reaction mixture to obtain a crude product of the target compound M18-1, wherein the crude product is directly used for the next reaction. Step 2: synthesis of intermediate compound M18-2

The crude 1-1 (5.64 g) was dissolved in DCM (80.00 mL) at room temperature, and TEA (24.11 mL) and dimethylamine hydrochloride (5.66 g) were added in ice bath and stirred at room temperature for 1h after the addition. Water was added to the reaction mixture, extraction was performed, and the organic phase was dried and concentrated. The concentrate was purified by a silica gel column to give the title compound M18-2 (5.4 g, yield 91%).

Step 3: synthesis of intermediate compound M18-3

Compound 18-2 (5.4 g) was dissolved in THF (120 mL) at room temperature, cooled to-20deg.C, and LiAlH was added in portions ₄ (4.0 g), the mixture was slowly returned to room temperature after the completion of the addition, and stirred for 3 hours. After the reaction was completed, 4.0g of water, 4.0g of 15% aqueous NaOH solution and 12.0g of water were slowly added in this order under ice bath conditions, and stirring was continued for 1 hour. The mixture was filtered and the filtrate was concentrated. Purification of the concentrate on a silica gel column (MeOH: dcm=0-30%) afforded the title compound M18-3 (2.5 g, 61%). ESI-MS m/z 130[ M+H ]] ⁺ 。

Step 4: synthesis of intermediate compound M18-4

DMSO (2.34 g) was dissolved in DCM (30 mL) at room temperature, oxalyl chloride (1.9 g) was added dropwise thereto at-78℃and stirred for 30min. A solution of compound M18-3 (1.3 g) in DCM (10 mL) was added dropwise thereto at-78deg.C and stirred for 1h. Triethylamine (3.1 g) was added dropwise thereto at-78℃and slowly returned to room temperature after completion of the addition. Adding water into the reaction solution, extracting, separating liquid, and separating water phase. The organic phase was concentrated to give crude product of the target compound M18-4. Step 5: synthesis of intermediate Compound M18

The crude product of the above compound M18-4 and potassium carbonate (2.8 g) were dissolved in methanol (10 mL) at room temperature, and stirred at room temperature for 15min. Dimethyl (1-diazo-2-oxopropyl) phosphonate (2.5 g) was added dropwise and stirred overnight at room temperature after addition. Toluene and water are added into the reaction liquid for extraction, the liquid is separated, and the organic phase is washed once by saturated potassium carbonate solution and dried by anhydrous sodium sulfate, thus obtaining crude product of the target compound M18.

Synthesis of intermediate compound M19:

reference to the synthetic preparation of intermediate M18, intermediate compound M19 is obtained. ESI-MS m/z 162[ M+H ]] ⁺ 。

Synthesis of intermediate compound M20:

step 1: synthesis of Compound M20-1

THF (80.00 mL), benzyl alcohol (1.60 g), was added to the flask at room temperature, cooled to 0deg.C, naH (0.59 g) was added, the reaction was completed at room temperature for 0.4h, cooled to 0deg.C, and Compound M2 (5.30 g) was added, and the reaction was completed at room temperature for 1h. The reaction mixture was quenched with water, extracted with EA, and the organic phase was dried and concentrated. The concentrate was purified by silica gel column (pe.about.pe: ea=1:1) to give the title compound M20-1 (5.60 g, yield 90.51%) as a yellow oil. ESI-MS m/z 500[ M+H ]] ⁺ 。

Step 2: synthesis of Compound M20-2

The compound M20-1 (1.90 g), 2- [6- (methoxymethoxy) -8- (4, 5-tetramethyl-1, 3, 2-dioxabenzaldehyde-2-yl) naphthalen-1-yl, was added to a reaction flask at room temperature ]Ethynyl tripropan-2-ylsilane (2.82 g), spospdg 2 (0.27 g), K ₃ PO ₄ (2.42 g) was dissolved in a mixed solution of 1,4-dioxane (30.00 mL) and water (5.00 mL), and reacted at 95℃for 10 hours under nitrogen atmosphere after nitrogen substitution. EA and water were added to the reaction mixture to extract, the organic phases were dried, and concentrated. The concentrate was purified by silica gel column (pe.about.pe: ea=3:1) to give the title compound M20-2 (1.80 g, yield 39.85%) as a yellow oil. ESI-MS m/z 902[ M+H ]] ⁺ 。

Step 3: synthesis of Compound M20-3

M20-2 (1.80 g), methanol (25.00 mL) and Pd/C (0.92 g) were added to the flask at room temperature, and the mixture was reacted at room temperature under hydrogen protection for 12 hours after hydrogen substitution. The reaction mixture was filtered, the filter residue was washed with methanol, the filtrate was concentrated and purified by column to give the title compound M20-3 (0.60 g, 37.38%) as a yellow solid.ESI-MS m/z:812[M+H] ⁺ 。

Step 12: synthesis of Compound M20

M20-3 (600.00 mg), dried DCM (8.00 mL) and DIEA (0.40 mL) were added to the reaction flask at room temperature, and the reaction mixture was cooled to-10deg.C to give Tf ₂ O (0.16 mL) was dissolved in DCM and added slowly to the reaction mixture, and the reaction was resumed at room temperature for 2h. The reaction solution was concentrated directly, and the concentrate was purified by a silica gel column (pe.about.pe: ea=8:1) to give the objective compound M20 (480.00 mg, 67.91%) as a yellow solid. ESI-MS m/z 944[ M+H ] ] ⁺ 。

Synthesis of intermediate M21:

step 1: synthesis of Compound M21-1

4-amino-2, 6-dichloropyridine (63 g) was added to a 1L three-necked flask, and 440mL of ACN and 180mL of water were added. The temperature was raised to 45℃and SelectFluor (164 g) was added, the reaction exothermed and allowed to cool naturally in air. After 10min of reaction, the reaction was quenched by addition of saturated sodium sulfite solution in ice bath, extracted 3 times with EA, the organic phases were collected and combined, dried over anhydrous sodium sulfate, concentrated, and the concentrate was purified by column chromatography (PE: ea=10:1) to give the title compound M21-1 (42.6 g). ESI-MS m/z 163.9[ M+H ]] ⁺ 。

Step 2: synthesis of Compound M21-2

M21-1 (54 g) was added to a 1L three-necked flask, 500mL of ACN was added, and NIS (80 g) and TsOH (5.1 g) were added. After reaction for 1h at 70℃2.5L of water were added dropwise, filtered and the filter cake was dissolved with EA. After separation, the organic phase was concentrated to give the objective compound M21-2 (94 g). ESI-MS m/z 307.2[ M+H ]] ⁺ 。

Step 3: synthesis of Compound M21-3

M21-2 (88 g) was added to a 1L three-necked flask, 500mL of DMF was added, and CuCN (31 g) was added. After the temperature is raised to 125 ℃ for reaction for 16 hours, 2L of water is added dropwise, the mixture is added into 2L of water, the mixture is filtered, and filter cakes are washed by water and dissolved by EA. The aqueous phase was extracted 2 times with EA, and the organic phases were collected and combined, and concentrated to give the desired product M21-3 (55.4 g). ES (ES) I-MS m/z:207.0[M] ^- 。

Step 4: synthesis of Compound M21-4

M21-3 (88 g) was added to a 250mL three-necked flask, 83mL concentrated sulfuric acid and 9mL water were added. After 16h of reaction at 60 ℃, 70mL of concentrated sulfuric acid and 5mL of water are supplemented, and the reaction is continued for 24h. Cooling to 0 ℃, slowly pouring the reaction solution into 2L of ice water, filtering, washing a filter cake with water, and dissolving EA. The filtrate was extracted twice with EA, the organic phases were collected and combined, and after concentration, PE: ea=1: 1 pulping. Filtration gave the desired product M21-4 (59 g). ESI-MS m/z 224.1[ M+H ]] ⁺ 。

Step 5: synthesis of Compound M21-5

M21-4 (58 g) was added to a 1L three-necked flask, 650mL of THF was added, and after heating to 40℃NaH (16 g) was added in slow portions. After stirring for 10min, the temperature was raised to 60℃and N, N' -thiocarbonyldiimidazole (69 g) was slowly added. After 1h of reaction, the reaction was quenched by addition of saturated ammonium chloride solution, diluted hydrochloric acid was added dropwise to adjust ph=4-5, and THF was removed by spinning. The mixture was filtered, and the filter cake was washed with water, dissolved in methanol, and the organic phase was concentrated to give crude M21-5 (85 g). ESI-MS m/z 267.0[ M+H ]] ⁺ 。

Step 6: synthesis of Compound M21

M21-5 (74 g) was added to a 1L three-necked flask, 750mL of ACN was added, methyl iodide (22 mL) was added, and an aqueous solution (100 mL) of sodium methoxide (23 g) was added. After 15min of reaction at room temperature, the reaction solution was added to 3.5L of water, ph=4-5 was adjusted by adding dilute hydrochloric acid, filtered, and the cake was treated with PE: ea=2: 1, pulping the mixed solvent. Filtering to obtain a filter cake which is the target product M21 (42.2 g). ESI-MS m/z 281.2[ M+H ] ] ⁺ 。

Synthesis of intermediate compound M22:

step 1: synthesis of Compound M22-1

4-bromo-2-fluoroaniline (11.0 g), potassium carbonate (20.18 g) and KI (9.7 g) were added to NMP (110.0 mL) respectively, and 1- (chloromethyl) -4-methoxybenzene (16.24 mL) was added dropwise thereto and stirred at room temperature for 7 hours. Pouring the reaction solution into H ₂ O (120 mL) was extracted 2 times with MTBE (60 mL) (three layers were separated, the upper layer was taken), and the organic layer was washed twice with saturated saline and dried over anhydrous sodium sulfate. The crude product was slurried with PE (60 ml) for 1 hour. Filtering, washing the filter cake with PE, and drying to obtain the compound M22-1 (17.68 g,70.33% yield). ESI-MS m/z=430 [ M+H ]] ⁺ 。

Step 2: synthesis of Compound M22-2

TMP (20.21 mL) was added to THF (170.00 mL), N at-10deg.C ₂ Three times of replacement, N ₂ n-BuLi (2.5M/n-hexane) (47.91 mL) was added dropwise under protection, stirring was carried out at-10℃for 10 minutes, the reaction was cooled to-60℃and a solution of M22-1 (17.18 g) in THF (40.00 mL) was added dropwise and stirred for 0.5 hour. DMF (30.90 mL) was then added quickly and stirred for 10 min. Pouring saturated NH into the reaction kettle ₄ Cl (200 mL) and then EA (50 mL) were used for extraction 2 times, and the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Concentrated under reduced pressure, and the crude product was slurried overnight with PE/mtbe=5/1 (60 ml). Filtration, washing the filter cake with PE/MTBE=5/1 and drying to obtain compound M22-2 (16.59 g, yield 90.66%). ¹ H NMR(500MHz,CDCl ₃ )δ10.34(d,J＝0.8Hz,1H),7.19(dd,J＝8.7,1.4Hz,1H),7.17-7.13(m,4H),6.86-6.80(m,5H),4.23(s,4H),3.78(s,6H)。ESI-MS m/z＝458[M+H] ⁺ 。

Step 3: synthesis of Compound M22-3

Compound M22-2 (16.09 g), cuI (1.34 g) was added to DMF (80.00 mL), N ₂ Three times of replacement, N ₂ The reaction was allowed to warm to 80℃under protection, and then methyl fluorosulfonyl difluoroacetate (14.97 mL) was added dropwise thereto, followed by stirring at 100℃for 1.5 hours. The mixture was filtered (celite), and the cake was washed 3 times with MTBE (50 ml), and the filtrate was washed 2 times with saturated brine. The emulsion was extracted with EA (40 mL), and the organic layer was washed 3 times with saturated saline. Purification by column chromatography (PE/EA, 10-15% EA) gave compound M22-3 (13.11 g, yield 83.46%). ¹ H NMR(500MHz,CDCl ₃ )δ10.43(q,J＝1.8Hz,1H),7.32(d,J＝8.6Hz,1H),7.17-7.13(m,4H),6.95(t,J＝8.5Hz,1H),6.87-6.82(m,4H),4.38(s,4H),3.79(s,6H)。ESI-MS m/z＝448[M+H] ⁺ 。

Step 3: synthesis of Compound M22-4

NaH (1.12 g) was added in portions to THF (50.00 mL), N ₂ The temperature was lowered to 0℃and methyl acetoacetate (3.01 mL) was added dropwise, followed by stirring for 10 minutes and n-BuLi (2.5M/n-hexane) (11.18 mL) was added dropwise and stirring was continued for 10 minutes. A solution of Compound M22-3 (5.00 g) in THF (5.00 mL) was added dropwise thereto at a temperature lowered to-15℃and stirred for 0.5 hours. Pouring the reaction solution into saturated NH ₄ Cl (100 ml) solution. The mixture was extracted with EA (40 ml) 2 times, and the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Purification by column chromatography (PE/EA, 30% EA) gave compound M22-4 (4.95 g, yield 78.60%). ESI-MS m/z=564 [ M+H ]] ⁺ 。

Step 4: synthesis of Compound M22-5

Compound M22-4 (4.85 g) was added to DCM (50.00 mL) and N, N-dimethylformamide dimethyl acetal (2.29 mL). N (N) ₂ Under protection, stirring at room temperature for 12 hours, LCMS monitored intermediate formation. Boron trifluoride diethyl etherate (1.95 mL) was added dropwise and stirred for 75 min after cooling to 0deg.C. Pouring saturated NH ₄ Cl solution (80 ml) and extracted 2 times with DCM (30 ml), and the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Purification by column chromatography (PE/EA, 35% EA) afforded Compound M22-5 (3.95 g, 80.02% yield). ESI-MS m/z=574 [ M+H ]] ⁺ 。

Step 5: synthesis of Compound M22-6

Compound M22-5 (3.75 g) was added to THF (40.00 mL) at N ₂ Lithium tri-sec-butylborohydride (7.85 mL,1.00 mol/L) was added dropwise at-65℃under protection, and stirring was continued for 0.5 hour. Pouring the reaction solution into saturated NH ₄ Cl (80 mL) and then EA (30 mL) were used for extraction 2 times, and the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Purification by column chromatography (PE/EA, 20% EA) gave Compound M22-6 (2.11 g, 56.07% yield). ESI-MS m/z=576 [ M+H ]] ⁺ . Step 6: synthesis of Compound M22-7

Compound M22-6 (2.01 g) was added to EtOH (40.00 mL) and H ₂ O (4.00 mL) was added 2-methyl-2-mercaptosulfuric acid urea (3.29 g) and Na ₂ CO ₃ (1.30 g). The reaction was stirred for 10 hours at 50 ℃. The reaction mixture was concentrated under reduced pressure, and then EA (30 ml) and H were added ₂ O (20 mL) was extracted twice, and the organic layer was washed with saturated brine and thenDried over anhydrous sodium sulfate. Purification by column chromatography (PE/EA, 45-50% EA) afforded Compound M22-7 (794 mg, 36.93% yield). ESI-MS m/z=616 [ M+H ]] ⁺ 。

Step 7: synthesis of Compound M22-8

Compound M22-7 (744.00 mg) was added to DCM (10.00 mL), followed by DIEA (1.00 mL), dropwise added Tf in ice bath ₂ O (0.61 mL). Stirred at 0℃for 0.5 h. Pouring the reaction solution into saturated NH ₄ Cl (15 ml), then extracted 2 times with DCM (15 ml), and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give Compound M22-8 (900 mg). ESI-MS m/z=748 [ M+H ]] ⁺ 。

Step 8: synthesis of Compound M22-9

Compound M22-8 (900.00 mg), M11 (604 mg) was added to DMF (10.00 mL) followed by DIEA (0.99 mL). Raise to 50℃and stir for 0.5 hours. EA (3 0 ml) and H ₂ O (30 mL) was extracted twice, and the organic layer was washed 3 times with saturated brine and dried over anhydrous sodium sulfate. Purification by column chromatography (PE/EA, 25% EA) gave compound M22-9 (520 mg, yield 55.8%). ESI-MS m/z=808 [ m+h ]] ⁺ 。

Step 9: synthesis of Compound M22

Compound M22-9 (500.00 mg) was added to DCM/dichloromethane (7.00 mL) and M-CPBA (170 mg, 85%) was added in portions and stirred at room temperature for 1.5 hours. LCMS monitored completion of the reaction. DCM (20 mL) and saturated NaHCO were added to the reaction ₃ The solution was washed 2 times, and the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Concentration gave 480mg of compound M22, crude which was used directly in the next step. ESI-MS m/z=824 [ m+h ]] ⁺ 。

Preparation of intermediate M23:

1) Preparation of L- (-) -di-p-methylbenzoyl tartrate of M23 Compound

Weighing 11g M9, adding 330ml of acetonitrile/water (30/1, volume ratio) mixed solvent into a reaction kettle, and stirring at 50 ℃ to dissolve to obtain M9 solution; 26.5g of L- (-) -di-p-methylbenzoyl tartaric acid was weighed into a round bottom flask, and 330ml of ethyl acetate solvent was added to prepare an acid solution. At the system temperature of 50 ℃, slowly dripping the acid liquor into the M9 solution, dripping 330ml of ethyl acetate into the reaction solution after the acid liquor dripping is finished, slowly cooling to 10 ℃ after the dripping is finished, and curing for half an hour at 10 ℃. The sample was suction filtered and the filter cake was dried under vacuum overnight at 40℃to give 13.9g of crude (S) -DPLT salt (solid isomer ratio 9.2%: 90.8%). The crude (S) -DPLT salt was recrystallized from ethanol/acetonitrile (1/1, volume ratio 1:1) to give 11.6g of (S) -DPLT salt M23-1 (isomer ratio 0.8%: 99.2%).

Single crystal cultivation and identification: weighing compound M23-1, dissolving with 1ml butanone/water (19/1, v/v) mixed solvent, transferring the clarified sample into butanone atmosphere, and performing gas-liquid diffusion overnight to obtain single crystal sample, wherein the single crystal sample is shown in figure 2 and the single crystal structure analysis is shown in figure 3, and the single crystal structure information table is shown below.

2) Preparation of M23

Weighing 20.0g of M23-1 (the R/S isomer ratio is 0.8 percent to 99.2 percent) into a reaction kettle, adding 200ml of dichloromethane/methanol (10/1, volume ratio), and stirring to dissolve; 160ml of purified water is added into the reaction liquid, 1M sodium hydroxide is added into the reaction liquid under stirring, the pH of the water phase is regulated to 11-12, and the mixture is stirred for half an hour and then separated; the aqueous phase is extracted by separating liquid after stirring for 20 minutes with 200ml of methylene dichloride/methanol (10/1, volume ratio) solvent, and repeated for 2 times; the organic phases were combined, washed with 200ml of pure water, then with 200ml of saturated brine, finally the organic phase was dried over anhydrous sodium sulfate, the dried organic phase was filtered, the filtrate was concentrated, and after concentration, 2 times of evaporation with ACN set, 5.17g of (S) - (2-methyltetrahydro-1H-pyrrolizine-7 a (5H) -yl) methanol M23 was obtained, and the R/S isomer ratio was 0.8%:99.2%.

Preparation example 1: synthesis of the Compound 4- (4- (1R, 5S) -3, 8-diazabicyclo [3.2.1] oct-3-yl) -2- (2- (difluoromethylene) tetrahydro-1H-pyrrolin-7 a (5H) -yl) methoxy) -8-fluoropyridine [4,3-d ] pyrimidin-7-yl) -5-ethynyl-6-fluoronaphthalen-2-ol

Step 1: synthesis of Compound 1-1

Compound M2 (0.6 g) and sodium hydride (0.11 g) were dissolved in anhydrous THF (2 mL) at room temperature, and a THF solution of compound M1 (0.53 g) was added dropwise at room temperature to react at room temperature for 0.5 hours. After the completion of the reaction, the solution was added dropwise to a saturated ammonium chloride solution at 0℃and then the organic layer was separated by adding EA, followed by washing with saturated brine (500 mL), drying over anhydrous sodium sulfate, and concentration. The concentrate was purified by column chromatography (EA: dcm=0-40%) to give compound 1-1 (0.54 g,66% yield). ESI-MS m/z=581.1 [ m+h ] ] ⁺ 。

Step 2: synthesis of Compounds 1-2

Compound 1-1 (0.3G), cataCXium APd G3 (40 mg), ((2-fluoro-6- (methoxymethoxy) -8- (4, 5-tetramethyl-1, 3, 2-dioxabenzaldehyde-2-yl) naphthalen-1-yl) ethynyl) triisopropylsilane (0.53G) and potassium phosphate (0.33G) were dissolved in THF (6 mL) and water (1 mL) at room temperature under nitrogen atmosphere, and the mixture was heated to 70℃to react for 3 hours, after the completion of the reaction, an organic layer was obtained by adding EA and separating, washing with saturated brine, drying over anhydrous sodium sulfate, and concentrating. The concentrate was purified by column chromatography (EA: dcm=0-30%) to give compound 1-2 (0.31 g,63% yield). ESI-MS m/z=931.1 [ m+h ]] ⁺ 。

Step 3: synthesis of Compounds 1-3

Compound 1-2 (0.31 g) was dissolved in DMF (3 mL), cesium fluoride (0.5 g) was added in portions at room temperature, after the reaction was completed, 15mL of water was added dropwise to the reaction liquid, a large amount of solid was precipitated, and the solid was obtained by filtration and dried to obtain compound 1-3 (0.31 g,76% yield). ESI-MS m/z=775.1 [ m+h ]] ⁺ 。

Step 4: synthesis of Compound 1

Compounds 1 to 3 (0.31 g) were dissolved in DCM (3 mL) at room temperature, trifluoroacetic acid (1 mL) was added thereto, after the completion of the reaction, the reaction solution was added dropwise to a protected sodium bicarbonate solution at 0℃and DCM was added to separate the organic layer, which was washed with saturated brine once, dried over anhydrous sodium sulfate, and concentrated. The concentrate was purified by Pre-HPLC to give compound 1 (31 mg,13% yield). ¹ H NMR(500MHz,DMSO-d ₆ )δ10.15(s,1H),9.04(s,1H),7.97(dd,J＝9.2,5.9Hz,1H),7.46(t,J＝9.0Hz,1H),7.39(d,J＝2.6Hz,1H),7.17(t,J＝2.3Hz,1H),4.47(d,J＝12.4Hz,1H),4.30(d,J＝12.2Hz,1H),4.14(dd,J＝10.6,4.2Hz,1H),4.08(dd,J＝10.6,2.3Hz,1H),3.93(dd,J＝4.0,1.0Hz,1H),3.68–3.60(m,2H),3.55(s,3H),3.30(s,1H),3.00(dq,J＝10.0,4.7,4.1Hz,1H),2.63(d,J＝14.7Hz,1H),2.56(q,J＝8.2Hz,1H),2.41(d,J＝15.8Hz,1H),1.97(td,J＝10.9,10.1,6.9Hz,1H),1.91–1.82(m,1H),1.78(q,J＝7.5Hz,2H),1.65(s,4H)。 ¹⁹ F NMR(471MHz,DMSO-d ₆ )δ-90.97(dd,J＝64.9,20.6Hz,1F),-91.46(dd,J＝64.9,38.2Hz,1F),-110.76(d,J＝3.8Hz,1F),-140.20(d,J＝3.1Hz,1F).

Preparation example 2: synthesis of the Compound 4- (4- (1R, 5S) -3, 8-diazabicyclo [3.2.1] oct-3-yl) -2- (2- (difluoromethylene) tetrahydro-1H-pyrrolin-7 a (5H) -yl) methoxy) -8-fluoro-5-isopropoxypyridin [4,3-d ] pyrimidin-7-yl) -5-ethynyl-6-fluoronaphthalen-2-ol

Step 1: synthesis of Compound 2-1

Isopropanol (1.23 ml) was dissolved in tetrahydrofuran (30 ml), naH (0.64 g) was added, and the reaction was carried out at room temperature for 10 minutes, the temperature was lowered to 0℃and 5, 7-dichloro-8-fluoro-2- (methylthio) pyridine [4,3-d ] was added]Pyrimidine-4-ol was removed to room temperature and reacted for 1h. After completion of the reaction, the reaction was quenched with saturated ammonium chloride, extracted twice with EA, washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to give compound 2-1 (2.06 g). ESI-MS m/z 304.26[ M+H ]] ⁺ 。

Step 2: synthesis of Compound 2-2

Compound 2-1 (2.06 g) and tert-butyl (1R, 5S) -3, 8-diazabicyclo [3.2.1]Octane-8-carboxylate (1.73 g) was dissolved in DMF (20 ml), DBU (4.13 g) was added thereto, pyBOP (5.29 g) was finally added thereto, and reacted at room temperature for 10 minutes. After the reaction was completed, water (100 ml) was slowly added dropwise to the reaction solution, a large amount of solids was precipitated, the obtained solid was filtered, dissolved with EA, dried over anhydrous sodium sulfate, concentrated, and the concentrate was purified by column chromatography to give compound 2-2 (1.35 g, yield 40.0%). ESI-MS m/z 498.11[ M+H ] ] ⁺ 。

Step 3: synthesis of Compound 2-3

The compound 2-2 (500 mg), ((2-fluoro-6- (methoxymethoxy) -8- (4, 5-tetramethyl-1, 3, 2-dioxabenzaldehyde-2-yl) naphthalen-1-yl) ethynyl) triisopropylsilane (705.54 mg), potassium carbonate (416.29 mg), [1,1' -bis (diphenylphosphine) ferrocene]Palladium dichloride dichloromethane complex (81.99 mg) was dissolved in 1, 4-dioxane (8.00 ml) and water (2.00 ml), and reacted under nitrogen atmosphere at 130℃in a can-sealed condition for 1 hour. After the reaction was completed, the reaction mixture was diluted with EA and water, extracted with EA once, washed with saturated brine once, dried over anhydrous sodium sulfate, concentrated, and the concentrate was purified by column chromatography to give compound 2-3 (713 mg, yield 88.32%). ESI-MS m/z 805.49[ M+H ]] ⁺ 。

Step 4: synthesis of Compounds 2-4

2-3 (713 mg) was dissolved in methylene chloride (10.00 ml) and t-butyldimethylchlorosilane (668.22 mg) and imidazole (422.57 mg) were added to the solution, and the mixture was reacted at room temperature for 0.5h. After the reaction was complete, the reaction mixture was diluted with DCM and water, extracted once with DCM, washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and the concentrate purified by column chromatography to give compound 2-4 (770 mg yield 94.56%).

Step 5: synthesis of Compounds 2-5

Compound 2-4 (770 mg) was dissolved in DCM (10.00 mL), and mCPBA (578.80 mg) was added and reacted at room temperature for 0.5h. The reaction was diluted with DCM, quenched with saturated sodium bicarbonate, extracted once with DCM, washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated and the concentrate purified by column chromatography to give compound 2-5 (290 mg).

Step 6: synthesis of Compounds 2-6

M1 was dissolved in THF (3.00 mL), naH (17.17 mg) was added thereto, and compound 2-5 (170.00 mg) was added thereto and reacted at room temperature for 0.5h. The reaction was quenched with saturated ammonium chloride, extracted twice with EA, washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the concentrate was purified by Pre-TLC to give compound 2-6 (163 mg). ESI-MS m/z 530.53[ M/2+H ]] ⁺ 。

Step 7: synthesis of Compounds 2-7

Compound 2-6 (163.00 mg) was dissolved in DMF (3.00 mL), csF (701.32 mg) was added, and the mixture was reacted at room temperature for 6h. The reaction solution was diluted with EA and water, extracted once with EA, washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the concentrate was purified by Pre-TLC to give compound 2-7 (73 mg). ESI-MS m/z 789.11[ M+H ]] ⁺ 。

Step 8: synthesis of Compound 2

Compound 2-7 (73.00 mg) was dissolved in DCM (2.00 mL), and boron trifluoride diethyl etherate (0.23 mL) was added and reacted at room temperature for 0.5h. The reaction was added to a cold saturated sodium carbonate solution using DCM: meoh=10:1, twice, one time with saturated brine, dry over anhydrous sodium sulfate, filter, concentrate, and purify the concentrate by Pre-TLC to give compound 2 (37.9 mg). ESI-MS m/z 689.25[ M+H ] ] ⁺ 。 ¹ H NMR(500MHz,DMSO)δ10.14(s,1H),10.14(s,1H),7.97(dd,J＝9.2,5.9Hz,1H),7.97(dd,J＝9.2,5.9Hz,1H),7.46(t,J＝9.0Hz,1H),7.46(t,J＝9.0Hz,1H),7.37(d,J＝2.5Hz,1H),7.37(d,J＝2.5Hz,1H),7.20(s,1H),7.20(s,1H),5.37-5.21(m,1H),4.16-4.02(m,3H),3.87(d,J＝2.3Hz,1H),3.64(d,J＝14.4Hz,1H),3.45(dd,J＝45.7,19.5Hz,4H),3.05-2.91(m,1H),2.67-2.53(m,2H),2.40(d,J＝15.9Hz,2H),2.04-1.72(m,4H),1.45-1.65(m,3H),1.30(dd,J＝6.2,2.0Hz,4H)。

Preparation example 3: synthesis of the Compound 4- (4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] oct-3-yl) -2- ((2- (difluoromethylene) tetrahydro-1H-pyrrolidin-7 a (5H) -yl) methoxy) -8-fluoropyrido [4,3-d ] pyrimidin-7-yl) -5-ethynyl-6-fluoronaphthalen-2-amine

Step 1: synthesis of Compound 3-1

Compound 1-1 (500 mg) and 6-fluoro-4- (4, 5-tetramethyl-1, 3, 2-dioxan-2-yl) -5- ((triisopropylsilyl) ethynyl) naphthalen-2-ol (882 mg) were dissolved in a mixed solvent of 1, 4-dioxane (10 mL) and water (2 mL), potassium phosphate (548 mg) was added, cataCXium A Pd G3 (63 mg) was added under nitrogen protection, and the mixture was heated to 70℃for 3 hours. Heating was stopped, cooled to room temperature, diluted with water (20 mL), extracted twice with dichloromethane (20 mL), the organic phases were combined, washed twice with water, dried over anhydrous sodium sulfate, filtered, concentrated and stirred, and the product was isolated by column chromatography (DCM: meoh=20:1) to yield 770mg of the title compound 3-1.

Step 2: synthesis of Compound 3-2

Compound 3-1 (720 mg) was dissolved in THF (20 mL), triethylamine (0.34 mL) and DMAP (10 mg) were added thereto, the temperature was lowered to 0℃under the protection of nitrogen, N-phenylbis (trifluoromethanesulfonyl) imide (580 mg) was dissolved in THF (4 mL), the mixture was dropped into the above-mentioned reaction solution, the reaction was allowed to proceed at room temperature for 10 minutes after the dropping, and then the mixture was stirred at room temperature for 2 hours. After LCMS monitored complete conversion of the starting material, the reaction was quenched in saturated sodium bicarbonate solution (50 mL), extracted twice with 50mL each time with DCM, the organic phases combined, washed twice with water, dried over anhydrous sodium sulfate, filtered, concentrated and stirred, and the product isolated by column chromatography (DCM: meOH) =25:1, isolating 686mg of the target compound 3-2.

Step 3: synthesis of Compound 3-3

Compound 3-2 (200 mg), benzophenone imine (71 mg) and cesium carbonate (192 mg) were dissolved in 1, 4-dioxane (3 mL), and BINAP (12 mg) and palladium acetate (4.4 mg) were added under nitrogen atmosphere, and the mixture was reacted at 100℃for 2 hours. After the reaction was completed, water (10 mL) was added to dilute, DCM was extracted twice, 15mL each time, the organic phases were combined, washed twice, dried over anhydrous sodium sulfate, filtered, concentrated and stirred, and the product (DCM: meOH) =50:1 was separated by column chromatography to give 230mg of the target compound 3-3.

Step 4: synthesis of Compounds 3-4

Compound 3-3 (200 mg) was dissolved in a mixed solution of hydrochloric acid solution (5 mL,2 mol/L) and THF (10 mL), and stirred at room temperature for 15min. Quenching the reaction solution in saturated carbonic acid solution, extracting with DCM twice, 15mL each time, combining the organic phases, washing twice, drying the organic phase with anhydrous sodium sulfate, filtering, concentrating to obtain the compound 3-4, and directly adding the compound into the next reaction without purification.

Step 5: synthesis of Compound 3-5

The above compound 3-4 was dissolved in DMF (3 mL), cesium fluoride (48 mg) was added, and the mixture was stirred at room temperature for 15min. Water (10 mL) was added, DCM was extracted twice, 15mL each time, the organic phases were combined, washed twice, dried over anhydrous sodium sulfate, filtered, concentrated and spun-dried to give crude product which was directly added to the next reaction without purification.

Step 6: synthesis of Compound 3

The crude product was dissolved in DCM (3 mL), and boron trifluoride diethyl etherate (0.12 mL) was added dropwise at room temperature under nitrogen blanket, and stirred at room temperature for 30min. The reaction was quenched in saturated carbonic acid solution, extracted twice with 15ml each time with DCM, the organic phases combined, washed twice with water, dried over anhydrous sodium sulfate, filtered, concentrated and isolated by pre-TLC (DCM: meoh=100:12) to give compound 3 (22.3 mg, 99.09% purity). ESI-MS m/z 630.5[ M+H ]] ⁺ 。 ¹ H NMR(500MHz,DMSO)δ9.06(s,1H),7.77(dd,J＝9.2,5.9Hz,1H),7.33(t,J＝9.0Hz,1H),7.07-7.01(m,2H),5.65(s,2H),4.58(d,J＝13.3Hz,1H),4.40(d,J＝12.9Hz,1H),4.20-4.06(m,2H),3.86-3.84(m,1H),3.82-3.62(m,3H),3.04-2.96(m,1H),2.70-2.54(m,2H),2.46-2.32(m,2H),2.04-1.94(m,2H),1.91-1.70(m,4H),1.49-1.18(m,4H)。

Preparation example 4: compound 2-amino-4- ((1 r,13 ar) -11-chloro-9-fluoro-7- ((2 r,7 as) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) -1-methyl-1, 2,3,4,13 a-hexahydropyrazine [2',1': synthesis of 3,4] [1,4] oxazine [5,6, 7-des ] quinazolin-10-yl) -7-fluorobenzo [ b ] thiophene-3-carbonitrile

Step 1: synthesis of Compound 4-1

The compound 3-methylpyrazine-2-carboxylic acid (6.0 g) was dissolved in methanol (150 mL), cooled below 0deg.C in an ice bath, and slowly added with SOCl dropwise ₂ (15.5 g), and after completion of the dropping, reacted at room temperature for 12 hours. The reaction solution was concentrated, the concentrate was diluted with methylene chloride, saturated brine was washed with water, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The concentrate was dissolved in dichloromethane (1 mL), crystallized from petroleum ether (20 mL), filtered and dried to give compound 4-1 (4.5 g,68% yield). ESI-MS m/z 153[ M+H ] ] ⁺ 。

Step 2: synthesis of Compound 4-2

Compound 4-1 (4.5 g) was dissolved in absolute ethanol (20 mL) and PtO was added ₂ (0.13 g) H was introduced ₂ The reaction was stirred at 70℃for 12h. The reaction solution was concentrated to give compound 4-2 (3.6 g,77% yield) which was directly fed to the next step.

Step 3: synthesis of Compound 4-3

Compound 4-2 (3.6 g) was dissolved in tetrahydrofuran (20 mL) and LiAlH was added in portions under an ice bath ₄ (2.25 g), and stirred at room temperature for 1h. The reaction was monitored to be complete and 2mL of H was slowly added dropwise ₂ O, 2mL of 15% NaOH aqueous solution is added, and 6mLH is added ₂ O, stirring at room temperature for 0.5h, adding sodium sulfate for drying after stirring and quenching are completed, filtering to obtain filtrate, and spin-drying to obtain the compound 4-3 (2.9 g,98% yield) which is directly added into the next step.

Step 4: synthesis of Compound 4-4

Compound 4-3 (2.9 g) was dissolved in methanol (100 mL), DIEA (14.39 g) and di-tert-butyl dicarbonate (12.15 g) were added, stirred at room temperature for 2h, a large amount of starting material was monitored to remain, DMAP (0.27 g) and DIEA (5.75 g) were added, and stirred at room temperature for 1h. The reaction was monitored to completion, the reaction mixture was concentrated, the resulting product was dissolved in ethanol, and an aqueous solution (50 mL) containing sodium hydroxide (2.6 g) was added thereto, followed by reflux reaction for 4 hours. The reaction solution was cooled to room temperature, the solvent was concentrated, then diluted with water (50 mL), pH was adjusted to about 8 with 6M hydrochloric acid, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, suction filtered, and the filtrate was concentrated to give Compound 4-4 (4.0 g,78% yield) which was directly added to the next step.

Step 5: synthesis of Compound 4-5

Compound 4-4 (4.0 g) was dissolved in methylene chloride (20 mL), TBSCl (5.24 g) and imidazole (5.91 g) were added, and the mixture was stirred at room temperature for 1h. After the reaction is completed, water is added for extraction, saturated saline water is used for washing, anhydrous sodium sulfate is used for drying, suction filtration is carried out, and filtrate is concentrated. The concentrate was purified by column chromatography on silica gel (PE/ea=3/2) to give compound 4-5 (2.6 g,43% yield).

Step 6: synthesis of Compounds 4-6

Compound 4-5 (1 g) was dissolved in DMF (10 mL), cesium fluoride (1.32 g) was added, reacted at room temperature for 14 hours, after the reaction was completed, the reaction solution was added to water, extracted with ethyl acetate, backwashed with saturated sodium chloride solution, the organic phase was concentrated, acetonitrile (5 mL) was added, then concentrated, tetrahydrofuran (5 mL) was added, and concentrated to obtain compound 4-6, which was directly put into the next step.

Step 7: synthesis of Compounds 4-7

The above-mentioned compounds 4 to 6 were dissolved in tetrahydrofuran, sodium hydride (100 mg) was added thereto, and the mixture was stirred at room temperature for 30 minutes. Intermediate M10 (300 mg) was added, stirring was continued at room temperature for 30 minutes, after completion of the reaction, the reaction solution was added to an aqueous ammonium chloride solution, extracted with ethyl acetate, the organic phase was concentrated, and the concentrate was purified by silica gel column to give the objective compound 4-7 (102 mg,22% yield). 551[ M+H ] ESI-MS m/z ] ⁺ 。

Step 8: synthesis of Compounds 4-8

Compound 4-7 (102 mg) was dissolved in DMF (5 mL), DIEA (0.1 mL) was added at room temperature, pyBOP (192 mg) was added slowly in portions with stirring at room temperature, stirring at room temperature was completed for 30 minutes, after completion of the reaction, the reaction solution was added to water, extracted with ethyl acetate, backwashed with saturated sodium chloride solution, the organic phase was concentrated, and the concentrate was purified by column on silica gel to give the objective compound 4-8 (62 mg,63% yield). ESI-MS m/z 533[ M+H ]] ⁺ 。

Step 9: synthesis of Compounds 4-9

In a reaction flask was charged compound 4-8 (62 mg), (3-cyano-7-fluoro-4- (4, 5-tetramethyl-1, 3, 2-dioxan-2-yl) benzo [ b)]Thiophene-2-alkyl) carbamic acid tert-butyl ester (97 mg), pd (DPEPhos) Cl ₂ (16 mg), potassium phosphate (74 mg), potassium fluoride (20 mg), cyclopentylmethyl ether (5 mL), nitrogen gas was purged, and the reaction was carried out at 80℃for 14 hours. After the reaction was cooled, it was added to water, extracted with ethyl acetate, the organic phase was concentrated, and the concentrate was purified by silica gel column to give the objective compound 4-9 (75 mg,86% yield). ESI-MS m/z 745[ M+H ]] ⁺ 。

Step 10: synthesis of Compounds 4-10

Compound 4-9 (75 mg) was dissolved in methylene chloride (5 mL), m-chloroperoxybenzoic acid (35 mg) was added in portions at room temperature, stirred at room temperature for 30 minutes, after completion of the reaction, the reaction mixture was added to water, extracted with ethyl acetate, the organic phase was concentrated, and the concentrate was purified by silica gel column separation to give the objective compound 4-10 (55 mg,70% yield). 777[ M+H ] ESI-MS m/z ] ⁺ 。

Step 11: synthesis of Compounds 4-11

Alcohol ((2R, 7 aS) -2-fluorotetrahydro-1H-pyrrolizine-7 a (5H) -yl) methanol (33 mg) was dissolved in tetrahydrofuran (3 mL), sodium hydride (9 mg) was added, and the mixture was stirred at room temperature for 30 minutes. 4-10 (55 mg) was added thereto, stirring was continued at room temperature for 30 minutes, and after completion of the reaction, the reaction solution was added to an aqueous ammonium chloride solution, extracted with ethyl acetate, the organic phase was concentrated, and the concentrate was purified by silica gel column separation to give the objective compound 4-11 (35 mg,58% yield). ESI-MS m/z 856[ M+H ]] ⁺ 。

Step 12: synthesis of Compound 4

Compound 4-11 (35 mg) was dissolved in methylene chloride (3 mL), and trifluoroacetic acid (1.5 mL) was added thereto, followed by stirring at room temperature for 30 minutes. After the reaction was complete, the reaction solution was concentrated and separated using preparative HPLC to give compound 4 (5.9 mg,22% yield). 656[ M+H ] ESI-MS m/z] ⁺ 。 ¹ H NMR(500MHz,DMSO-d ₆ )δ8.08(d,J＝6.6Hz,2H),7.23(dt,J＝9.2,4.9Hz,1H),7.17-7.10(m,1H),5.47-5.16(m,1H),4.79(t,J＝14.3Hz,1H),4.43(ddd,J＝24.6,12.6,5.3Hz,1H),4.13-3.90(m,3H),3.08(d,J＝10.9Hz,2H),3.03(d,J＝8.5Hz,1H),2.88-2.77(m,2H),2.13(d,J＝9.3Hz,1H),2.07(q,J＝5.2,3.5Hz,1H),2.04-1.98(m,1H),1.77(dd,J＝11.9,5.6Hz,2H),1.34(t,J＝4.0Hz,1H),1.24(d,J＝7.4Hz,3H),1.08(q,J＝6.7,5.8Hz,3H),0.92(t,J＝7.1Hz,1H),0.88-0.80(m,1H)。

Preparation example 5: synthesis of Compound 3- (4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] oct-6-en-3-yl) -2- ((2- (difluoromethylene) tetrahydro-1H-pyrrolidin-7 a (5H) -yl) methoxy) -7, 8-dihydro-5H-pyran [4,3-d ] pyrimidin-7-yl) -4-fluoromethylaniline trifluoroacetate

Step 10: synthesis of Compound 5-1

Intermediate M1 (120 mg) was added to THF (3.00 mL) and t-Buona (70 mg) was added thereto, and the mixture was stirred at room temperature for 0.5 hours. A solution of Compound M22 (300 mg) in THF (1.00 mL) was then added. Stirred at room temperature for 0.5 hours. LCMS monitored completion of the reaction. Reaction with addition of saturated NH ₄ Cl (20 mL) was quenched, and after 2 times extraction with EA (25 mL), the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. Filtration, concentration and purification by column chromatography (DCM/MeOH, 5-10% MeOH) afforded compound 5-1 (150 mg, 40.07% yield). ESI-MS m/z=949 [ M+H ]] ⁺ 。

Step 11: synthesis of Compound 5

Compound 5-1 (150 mg) was added to DCM (3.00 mL) and TFA (3.00 mL) was added and stirred at room temperature for 0.5 h. Concentrating under reduced pressure, and preparing the crude product by HPLC to obtain the trifluoroacetate salt of Compound 5 (53 mg,55.1% yield, 95.05% purity). ¹ H NMR(500MHz,DMSO-d ₆ )δ10.71(s,1H),9.30(d,J＝10.2Hz,1H),9.21(d,J＝10.0Hz,1H),7.27(d,J＝8.6Hz,1H),6.84(t,J＝8.5Hz,1H),5.03(dd,J＝11.2,4.4Hz,1H),4.84(d,J＝13.8Hz,1H),4.78-4.60(m,2H),4.67(d,J＝13.8Hz,1H),4.48-4.42(m,3H),4.18(d,J＝14.2Hz,2H),4.13-4.07(m,3H),3.91(d,J＝14.6Hz,1H),3.70-3.52(m,3H),3.26-3.12(m,3H),2.86(td,J＝13.1,10.8,7.3Hz,2H),2.73(d,J＝16.1Hz,1H),2.24-1.88(m,7H),1.82(q,J＝11.2Hz,1H)。ESI-MS m/z＝609[M+H] ⁺ 。

Preparation example 6: synthesis of the Compound 3- (4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] oct-6-en-3-yl) -2- ((2-methylenetetrahydro-1H-pyrrolidin-7 a (5H) -yl) methoxy) -7, 8-dihydro-5H-pyrano [4,3-d ] pyrimidin-7-yl) -4-fluoromethylaniline

Step 1: synthesis of Compound 6-1

Intermediate M9 (100 mg) was dissolved in THF (2.50 mL), and t-Buona (60 mg) was added thereto and stirred at room temperature for 0.5 hours. A solution of Compound M22 (270 mg) in THF (1.00 mL) was then added. Stirred at room temperature for 0.5 hours. Adding saturated NH into the reaction solution ₄ Cl (20 mL) was quenched, and after 2 times extraction with EA (25 mL), the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified by column chromatography (DCM/MeOH, 5-10% MeOH) to give compound 6-1 (90 mg, 28.07% yield). ESI-MS m/z=913 [ M+H ] ] ⁺ 。

Step 2: synthesis of Compound 6

Compound 6-1 (90 mg) was dissolved in DCM (2.00 mL), TFA (2.00 mL) was added and stirred at room temperature for 0.5 h. Concentrating under reduced pressure, and purifying the crude product by Pre-TLC to give Compound 6 (32 mg, purity 98.5%, yield 56.7%). ¹ H NMR(500MHz,DMSO-d ₆ )δ7.25(d,J＝8.6Hz,1H),6.80(t,J＝8.5Hz,1H),5.25-5.17(m,2H),5.00(m,1H),4.80(d,J＝13.8Hz,1H),4.75-4.60(m,2H),4.63(d,J＝13.8Hz,1H),4.45-4.40(m,3H),4.15(d,J＝14.2Hz,2H),4.12-4.07(m,3H),3.90(d,J＝14.6Hz,1H),3.69-3.50(m,3H),3.23-3.10(m,3H),2.84(m,2H),2.67(m,1H),2.20-1.84(m,7H),1.80(m,1H)。ESI-MS m/z＝573[M+H] ⁺ 。

Preparation example 7: synthesis of the Compound 4- (4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] oct-3-yl) -6-chloro-8-fluoro-2- ((2R, 7 aS) -2-fluorotetrahydro-1H-pyrrolin-7 a (5H-yl) methoxy) quinazolin-7-yl) -2-amino-7-fluorobenzo [ b ] thiophene-3-carbonitrile

Step 1: synthesis of Compound 7-1

At room temperature, 3- (7-bromo-2, 6-dichloro-8-fluoro-quinazolin-4-yl) -3, 8-diazabicyclo [3.2.1] are added sequentially]Octane-8-carboxylic acid tert-butyl ester (100.00 mg), [ (2R, 8S) -2-fluoro-1, 2,3,5,6, 7-hexahydropyrrolizin-8-yl]Methanol (34.61 mg), cs ₂ CO ₃ (193.15 mg) DABCO (2.22 mg) to a mixed solution of DMF (1.00 mL) and THF/tetrahydrofuran (1.00 mL). Nitrogen bubbling, gas displacement, and stirring at room temperature overnight. The reaction solution was diluted with water, extracted with EA, and the organic layer was washed with saturated brine three times, dried and concentrated, and purified by column chromatography to give compound 7-1 (62.00 mg, yield 49.89%) as a pale yellow solid. ESI-MS m/z 628[ M+H ]] ⁺ 。

Step 2: synthesis of Compound 7-2

The compound 7-1 (20.00 mg), tert-butyl N- [ 3-cyano-7-fluoro-4- (4, 5-tetramethyl-1, 3, 2-dioxabenzaldehyde-2-yl) benzothien-2-yl, was reacted at room temperature]Carbamates (26.60 mg), pd (dtbpf) Cl ₂ (4.13mg)，K ₃ PO ₄ (16.88 mg) was added to a mixed solution of 1, 4-dioxane (1.00 mL) and water (0.10 mL) in this order. The nitrogen is replaced by gas and is placed in an oil bath at 90 ℃ for reaction. Diluting with water, extracting with DCM, drying and concentrating the organic phase, and separating and purifying the concentrate by Pre-TLC to obtain the title compound 7-2 (24.00 mg, yield 90.3%) as a pale yellow solid. ESI-MS m/z 840[ M+H ]] ⁺ 。

Step 3: synthesis of Compound 7

Compound 7-2 (48.00 mg) was added to a mixed solvent of trifluoroacetic acid (1.00 mL) and dichloromethane (1.00 mL) at room temperature, and stirred at room temperature for 30min. The reaction solution was concentrated, diluted with DCM, pH adjusted to weak base with saturated sodium bicarbonate in ice water bath, extracted with separated liquid, and concentrated by organic phase drying. The concentrate was purified by Pre-TLC to give the title compound 7 (22.00 mg, yield 60.19%) as a white solid. ESI-MS m/z 640[ M+H ]] ⁺ 。 ¹ H NMR(500MHz,MeOD)δ7.84(d,J＝8.2Hz,1H),7.24-7.15(m,1H),7.02(dd,J＝16.9,8.0Hz,1H),5.41-5.19(m,1H),4.57-4.39(m,1H),4.32-4.15(m,1H),3.69-3.53(m,1H),3.20(dd,J＝20.2,10.5Hz,1H),2.99(d,J＝22.1Hz,1H),2.37-2.10(m,1H),2.06-1.74(m,1H),1.45-1.21(m,1H)。

Preparation example 8: synthesis of the Compound 4- (4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] oct-3-yl) -6-chloro-8-fluoro-2- ((2-oxodihydro-1H-pyrrolin-7 a (5H) -yl) methoxy) quinazolin-7-yl) -2-amino-7-fluorobenzo [ b ] thiophene-3-carbonitrile

Step 1: synthesis of Compound 8-1

M9 (120.00 mg) and intermediate M17 (36.22 mg) were added to tetrahydrofuran (2.50 mL) at room temperature, followed by sodium t-butoxide (30.29 mg). Stirring at room temperature for 10 minutes. LCMS monitored completion of the reaction. Filtration, washing of the filter cake with DCM, and purification by column chromatography (DCM/MeOH, 5% MeOH) gave the title compound 8-1 (101 mg, 76.81% yield). ESI-MS m/z 834[ M+H ]] ⁺ 。

Step 2: synthesis of Compound 8

Compound 8-1 (55.00 mg) was added to dichloromethane (3.00 mL) at room temperature followed by TFA (0.80 mL). Stirring at room temperature for 1.5 hours. LCMS monitored completion of the reaction. Concentrating under reduced pressure, and concentrating the crude product with saturated NaHCO ₃ After adjusting to ph=8, extracting 2 times with DCM/MeOH (10/1), the organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, and concentrated, and the concentrate was prepared by Pre-HPLC to give compound 8 (16.4 mg, yield 38.80%, purity 98.9%). ¹ H NMR (500 MHz, deuterated methanol) delta 7.95 (d, j=1.6 hz, 1H), 7.22 (dd, j=8.3, 5.0hz, 1H), 7.06 (dd, j=9.4, 8.3hz, 1H), 5.28 (d, j=13.4 hz, 2H), 4.78-4.63 (m, 6H), 4.41-4.31 (m, 1H), 4.23 (s, 2H), 3.91 (dd, j=26.3, 13.9hz, 3H), 3.77 (dq, j=11.9, 5.9hz, 1H), 3.10-3.01 (m, 1H), 2.80 (d, j=16.5 hz, 1H), 2.38 (dp, j=18.3, 5.9hz, 1H), 2.28-2.13 (m, 4H), 1.31 (d, j=20.9 hz, 1H). ESI-MS m/z 634[ M+H ] ] ⁺ 。

Preparation example 9: synthesis of Compound 4- (4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octyl-3-yl) -6-chloro-2- ((2- (difluoromethylene) tetrahydro-1H-pyrrolin-7 a (5H) -yl) methoxy) -8-fluoroquinazolin-7-yl) -2-amino-7-fluorobenzo [ b ] thiophene-3-carbonitrile

Step 1: synthesis of Compound 9-1

The compound 2, 5-dioxodihydro-1H-pyrrolizine-7 a (5H) -carboxylic acid ethyl ester (500 mg) was dissolved in methanol (10 mL) at room temperature, thionyl chloride (0.69 mL) was added slowly at 0deg.C, and then warmed to room temperature for reaction for 1H. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, the residue was extracted with saturated aqueous sodium hydrogencarbonate (50 mL) and DCM (50 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated to give a crude product of compound 9-1, which was used in the next reaction without purification.

Step 2: synthesis of Compound 9-2

The crude product of the above compound 9-1 was dissolved in 20mL of anhydrous THF at room temperature, LAH (222 mg) was slowly added at 0℃and reacted at 70℃for 1h. Then cooling to 0 ℃, adding 222mL of water, adding 222mL of 15% strong sodium oxide aqueous solution, adding 666mL of water, reacting for 30min at room temperature, adding a proper amount of anhydrous sodium sulfate, reacting for 15min at room temperature, filtering, and spin-drying the filtrate to obtain the compound 9-2 (370 mg).

Step 3: synthesis of Compound 9-3

Intermediate M17 (900 mg) and compound 9-2 (357 mg) were dissolved in anhydrous THF (20 mL) at room temperature, and t-Buona (228 mg) was then added slowly and reacted at room temperature for 10min to completion. Quenched by adding water (50 mL) and extracted by adding EA (50 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give the title compound 9-3 (963 mg). ESI-MS m/z 882[ M+H ]] ⁺ 。

Step 4: synthesis of Compound 9-4

Compound 9-3 (800 mg) was dissolved in THF (2 mL) and 4M HCl 1, 4-dioxane (8 mL) at room temperature, reacted at 50℃for 10min, and solid was separated out, and filtered to give the hydrochloride (845 mg) of the objective compound 9-4. ESI-MS m/z 636[ M+H ]] ⁺ 。

Step 5: synthesis of Compound 9-5

Compound 9-4 (800 mg), (Boc) was added at room temperature ₂ O (1.37 g), DIPEA (1.6 g) and DMAP (31 mg) were dissolved in THF (12 mL) and DMF (3 mL) and reacted at room temperature for 10min. The reaction solution was slowly poured into saturated sodium bicarbonate (20 mL), stirred for 30min, then extracted with EA (50 mL) and water (30 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give the title compound 9-5 (568 mg). ¹ H NMR(500MHz,Chloroform-d)δ8.02(d,J＝1.5Hz,1H),7.79-7.67(m,1H),7.30(td,J＝5.5,4.9,2.4Hz,1H),7.15(t,J＝8.7Hz,1H),4.46-4.21(m,6H),3.66(d,J＝18.9Hz,2H),3.35(dtd,J＝10.1,5.1,2.9Hz,1H),3.26-3.13(m,1H),2.80(d,J＝18.8Hz,1H),2.74(dt,J＝9.8,7.9Hz,1H),2.46-2.36(m,1H),2.33-2.21(m,1H),2.03-1.97(m,2H),1.97-1.92(m,2H),1.92-1.73(m,4H),1.56(s,9H),1.53(s,9H)。ESI-MS m/z:836[M+H] ⁺ 。

Step 6: synthesis of Compound 9-6

Compound 9-5 (50 mg) and difluoromethyl (2-pyridyl) sulfone (116 mg) were dissolved in anhydrous DMF (3 mL) at room temperature, a solution of potassium tert-butoxide (67 mg) in DMF (1 mL) was added dropwise at-50℃for 2 hours at-40℃and, after completion of the reaction, a solution of saturated ammonium chloride (5 mL) was added dropwise at-50℃for 8 hours at-50℃and then reacted at room temperature, EA (10 mL) was added for dilution, washed once with water (20 mL), saturated brine (20 mL) was washed once, dried over anhydrous sodium sulfate and concentrated. The concentrate was purified by column chromatography (MeOH: dcm=0-10%) to give compound 9-6 (25 mg, 48% yield). ESI-MS m/z=870 [ m+h ] ] ⁺ 。

Step 7: synthesis of Compound 9

Compound 9-6 (22 mg) was dissolved in DCM (1 mL) at room temperature, TFA (1 mL) was added, and after the addition was completed, the mixture was stirred at room temperature for 30min. After the reaction was completed, the reaction solution was added dropwise to 10mL of saturated sodium bicarbonate solution, and the aqueous phase was extracted 3 times with 4mL of DCM. The organic phases were combined, washed once with 5mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated. The concentrate was purified by Pre-HPCL to give compound 9 (20.1 mg, yield 87%). ¹ HNMR(500MHz,)δ10.81(s,1H),9.32(s,1H),9.05(s,1H),8.14(s,2H),7.98(s,1H),7.26-7.15(m,2H),4.70-4.57(m,3H),4.40(d,J＝13.9Hz,1H),4.28-4.24(m,1H),4.23-4.06(m,4H),3.92(d,J＝14.0Hz,1H),3.73-3.70(m,1H),3.31(s,1H),2.97(d,J＝16.1Hz,1H),2.83(d,J＝15.5Hz,1H),2.23-2.01(m,4H),1.93-1.48(m,4H)。ESI-MS m/z＝670[M+H] ⁺ 。

Preparation example 10: synthesis of the Compound 4- (4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] oct-3-yl) -6-chloro-8-fluoro-2- ((2R, 7 aS) -2-fluorotetrahydro-1H-pyrrolin-7 a (5H-yl) methoxy) quinazolin-7-yl) -2-aminobenzo [ b ] thiophene-3-carbonitrile

Step 1: synthesis of Compound 10-1

Intermediate M15 (10.5 g), intermediate 7-1 (10.0 g), pd (DPEPhos) Cl were reacted at room temperature under nitrogen atmosphere ₂ (2.3 g) and potassium phosphate (10.1 g) were dissolved in toluene (150 mL), and the mixture was stirred at 100℃for 9 hours. After completion of the reaction, the reaction mixture was diluted with EA (300 mL), washed with water (100 mL) and saturated brine (100 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated. Purification of the concentrate by column chromatography (EA: pe=1-20%) gave compound 10-1 (8.5 g, 65% yield). ESI-MS m/z=822 [ m+h ] ] ⁺ 。

Step 2: synthesis of Compound 10

Compound 10-1 (8.5 g) was dissolved in DCM (80 mL) at room temperature, TFA (80 mL) was added dropwise at room temperature, and after addition was completed, stirring was performed at room temperature for 30min. After the completion of the reaction, the reaction solution was concentrated. Adding NH ₃ Neutralized with methanol (7M, 20 mL) and concentrated. The concentrate was purified by column chromatography ((7M NH) ₃ in MeOH): dcm=1-10%) to give the title compound 10 (5.4 g, 86% yield). ESI-MS m/z=622 [ m+h ]] ⁺ 。

Preparation example 11: synthesis of Compound 4- (4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octyl-3-yl) -8-fluoro-2- ((1- (pyrrolidin-1-ylmethyl) cyclopropyl) ethynyl) quinazolin-7-yl) -5-ethynyl naphthalen-2-ol

Step 1: synthesis of Compound 11-1

Compound M20 (80 mg), pd (PPh) ₃ ) ₂ Cl ₂ (11 mg), cuI (6 mg) and triethylamine (40 mg) were dissolved in a toluene solution of Compound M19, and the mixture was stirred at 50℃for 2 hours. Water and EA were added to the reaction solution, the mixture was extracted and separated, the organic phase was dried and concentrated, and the concentrate was purified by column chromatography (EA: pe=20 to 50%) to give the objective compound 11-1. Step 2: synthesis of Compound 11-2

Compound 11-1 (110 mg) was dissolved in THF (5 mL) at room temperature, TBAF (1M/THF, 0.12 mL) was added dropwise, and the mixture was stirred at room temperature for 30min. The reaction solution was concentrated, and the concentrate was purified by column chromatography (MeOH: dcm=0-200%) to give the objective compound 11-2 (80 mg, yield 90%). ESI-MS m/z 730[ M+H ] ] ⁺ 。

Step 3:

the crude compound 11-2 was dissolved in DCM (5 mL) at room temperature, to which trifluoroacetic acid (3 mL) was added dropwise and stirred at room temperature for 30min. The reaction solution was concentrated to obtain a crude product of compound 11. ESI-MS m/z 585[ M+H ]] ⁺ 。

Example 12: synthesis of the Compound 4- (4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] oct-3-yl) -8-fluoro-2- ((2-methyltetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) pyrido [4,3d ] pyrimidin-7-yl) -5-ethynyl-6-fluoronaphthalen-2-amine

Step 1: synthesis of Compound 12-1

Intermediate M9 (430 mg) was dissolved in anhydrous THF (20 mL), nitrogen blanketed, sodium hydride (220 mg) was added, intermediate M2 (1 g,2.33 mmol) was added, and stirred at room temperature for 4h. The reaction solution was quenched in anhydrous ammonium chloride (40 mL), extracted twice with ethyl acetate, 40mL each time, the organic phases were combined, washed twice with water, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified by column chromatography (DCM: meoh=40:1) to give 760mg of compound 12-1.

Step 2: synthesis of Compound 12-2

To a mixed solvent of 1, 4-dioxane (15 mL) and water (3 mL) was dissolved compound 12-1 (760 mg) and intermediate compound M13 (980 mg), potassium phosphate (888 mg) was added, cataCXium A Pd G3 (101 mg) was added under nitrogen protection, and the temperature was raised to 100℃for reaction for 3 hours. Heating was stopped, cooled to room temperature, diluted with water (40 mL), extracted twice with 40mL each time of dichloromethane, the organic phases were combined, washed twice with water, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrate was purified by column chromatography (DCM: meoh=20:1) to give 760mg of compound 12-2.

Step 3: synthesis of Compound 12-3

Compound 12-2 (760 mg) was dissolved in THF (20 mL), triethylamine (0.37 mL) and DMAP (11 mg) were added thereto, the temperature was lowered to 0℃under the protection of nitrogen, N-phenylbis (trifluoromethanesulfonyl) imide (638 mg) was dissolved in THF (4 mL), the mixture was dropped into the above-mentioned reaction solution, the reaction was allowed to proceed at room temperature for 10 minutes after the dropping, and then the mixture was stirred at room temperature for 1 hour. The reaction was quenched in saturated sodium bicarbonate solution (50 mL), extracted twice with 50mL each time with DCM, the organic phases were combined, washed twice with water, dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography (DCM: meoh=25:1) to give 635mg of compound 12-3.

Step 4: synthesis of Compound 12-4

Compound 12-3 (635 mg), benzophenone imine (234 mg) and cesium carbonate (631 mg) were dissolved in 1, 4-dioxane (15 mL), and BINAP (40 mg) and palladium acetate (15 mg) were added under nitrogen atmosphere, and the mixture was reacted at 100℃for 1 hour. Dilute with water (40 mL), extract twice with 50mL each time with DCM, combine the organic phases, wash twice with water, dry the organic phase over anhydrous sodium sulfate, filter, and concentrate. The concentrate was purified by column chromatography (DCM: meoh=50:1) to give 560mg of yellow oil 12-4.

Step 5: synthesis of Compound 12-5

12-4 (450 mg) was dissolved in a mixed solution of hydrochloric acid solution (5 mL,2 mol/L) and THF (10 mL), and stirred at room temperature for 15min. Quenching the reaction solution in saturated sodium carbonate solution, extracting with DCM for two times, combining 30ml each time, washing with water for two times, drying the organic phase with anhydrous sodium sulfate, filtering, concentrating to obtain crude product 12-5, and directly feeding the crude product into the next reaction without purification.

Step 6: synthesis of Compound 12-6

The crude product was dissolved in DMF (7 mL), cesium fluoride (1.3 g) was added, and stirred at 35℃for 30min. Dilute with water (30 mL), extract twice with 3mL each time with DCM, combine the organic phases, wash twice with water, dry the organic phase over anhydrous sodium sulfate, and concentrate. Concentrating to obtain crude product, and directly feeding into next reaction without purification.

Step 7: synthesis of Compound 12

The crude product is dissolved in DCM (6 mL), nitrogen is used for protection, boron trifluoride diethyl etherate (0.34 mL) is added dropwise at room temperature, the addition is completed, stirring is carried out for 10min at room temperature, the reaction solution is quenched in saturated carbonic acid solution, DCM is extracted twice, 30mL each time, the organic phases are combined, water washing is carried out twice, and the organic phase is dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by Pre-TLC (DCM: meoh=100:12) to give 12 (23.0 mg, purity 98.24%). [ M+H ]] ⁺ ＝594.45。 ¹ H NMR(500MHz,CDCl ₃ )δ8.96(s,1H),7.66(dd,J＝9.1,5.7Hz,1H),7.20(t,J＝8.9Hz,1H),7.09(dd,J＝15.6,2.4Hz,2H),4.93(s,2H),4.72–4.60(m,1H),4.55–4.43(m,1H),4.24–4.12(m,2H),3.92(s,2H),3.76–3.64(m,4H),3.58(dd,J＝12.3,4.8Hz,1H),3.28(d,J＝14.1Hz,1H),3.22–3.15(m,1H),2.86–2.75(m,2H),2.71–2.61(m,1H),2.40(d,J＝14.4Hz,1H),2.19(ddd,J＝17.4,11.9,7.2Hz,1H),1.97–1.87(m,2H),1.85-1.75(m,2H),1.72-1.54(m,4H)。

Example 18: synthesis of the Compound 4- (4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] oct-3-yl) -8-fluoro-5-methoxy-2- ((2-methylenetetrahydro-1H-pyrrolidin-7 a (5H) -yl) methoxy) pyridin [4,3-d ] pyrimidin-7-yl) -5-ethyne-6-fluoronaphthalen-2-ol

Step 1: synthesis of Compound 18-1

MeOH (1.80 mL) and anhydrous THF are respectively added into a reaction bottle, the temperature is reduced in an ice-water bath for 5min, naH (2.86 g) is slowly added in batches, after stirring for 5min, powder solid M21 (4.00 g) is added in batches, and the mixture is naturally heated to room temperature and 22 ℃ for stirring reaction for 10min; cooling in ice water bath for 5min, and dripping saturated NH ₄ Quenching with 100mL of Cl, adding 200mL of ethyl acetate, extracting, washing the water phase with ethyl acetate for 2 times, merging the organic phases, and evaporating under reduced pressure to obtain the target compound 18-1(4.23g)。ESI-MS m/z:276.0[M+H] ⁺ 。

Step 2: synthesis of Compound 18-2

The compound 18-1 (4.23 g), (1R, 5S) -3, 8-diazabicyclo [3.2.1 ] is added sequentially to the reaction flask at room temperature]After tert-butyl octane-8-carboxylate (3.91 g), DBU (11.47 mL) and anhydrous DMF (10.0 mL) were dissolved by stirring at room temperature, pyBOP (12.00 g) was added and the reaction was stirred at room temperature for 0.5h. 200mL of saturated NaCl-tap water and 200mL of EA were added to the reaction, the mixture was washed twice, the separated liquid was extracted, the organic phase was evaporated to dryness under reduced pressure, and the concentrate was purified by column chromatography (PE/EA=3:1) to give the objective compound 18-2 (2.57 g). ESI-MS m/z 470.3[ M+H ]] ⁺ 。

Step 3: synthesis of Compound 18-3

The microwave tube was charged with compound 18-2 (0.70 g), M13 (1.40 g), K, respectively ₂ CO ₃ (0.63g)，Pd(dppf)Cl ₂ (0.20 g) and 7mL of Dioxane/1mL of H ₂ After O, N ₂ After 2min of displacement, the reaction was carried out for 0.5h at 140℃with microwaves. DCM and H were added to the reaction solution ₂ O20 mL each, extract, wash the aqueous phase twice with DCM, concentrate the organic phase under reduced pressure, and purify the concentrate by column chromatography (PE/EA=4:1) to give the title compound 18-3 (1.23 g). 776.4[ M+H ] ESI-MS m/z] ⁺ 。

Step 4: synthesis of Compound 18-4

To the flask were added 18-3 (1.13 g), TBSCl (1.10 g), imidazole (0.69 g) and anhydrous DCM (15 mL), respectively, and the reaction was stirred at room temperature for 20min. 15mL of saturated NaCl-tap water was added to the reaction, the solution was separated by extraction, the aqueous phase was washed twice with 10mL of DCM, the organic phase was evaporated to dryness under reduced pressure, and the concentrate was purified by column chromatography (PE/EA=10:1) to give the objective compound 18-4 (1.06 g). ESI-MS m/z 890.3[ M+H ] ] ⁺ 。

Step 5: synthesis of Compound 18-5

To the flask were added 18-4 (1.00 g), mCPBA (0.47 g) and anhydrous DCM (15 mL), respectively, and the reaction was stirred at room temperature for 15min. To the reaction was added 20mL of saturated NaHCO ₃ The extract was separated, the aqueous phase was washed twice with 10mL DCM, the organic phase was evaporated to dryness under reduced pressure and the concentrate was purified by column chromatography (PE/ea=3:1) to give the title compound 18-5 (0.40 g). ESI-MS m/z 923.0[ M+H ]] ⁺ 。

Step 6: synthesis of Compound 18-6

To the reaction flask, M4 (0.11 g) and anhydrous THF (1 mL) were added, naH (0.25 g) was added slowly in portions, followed by stirring for 10min, 18-5 (0.33 g) was added, and the reaction was stirred at room temperature for 15min. Cooling the reaction solution with ice water bath for 5min, and dripping saturated NH ₄ Cl 20mL quenching, adding 20mL ethyl acetate, extraction, aqueous phase with ethyl acetate 2 times, combined organic phase, reduced pressure evaporation, concentrate column chromatography purification (DCM/NH) ₃ : meoh=20:1) to afford the title compound 18-6 (0.32 g). ESI-MS m/z 881.6[ M+H ]] ⁺ 。

Step 7: synthesis of Compound 18-7

To the reaction flask, compound 18-6 (0.32 g), csF (0.55 g) and anhydrous DMF (3 mL) were added, respectively, and the reaction was stirred at room temperature for 5.0h. To the reaction was added 20mL of saturated NaCl, the extract was separated, the aqueous phase was washed twice with 20mL of EA, the organic phase was evaporated to dryness under reduced pressure, and the concentrate was purified by column chromatography (DCM/NH) ₃ in MeOH=20:1) to give the title compound 18-7 (0.18 g). ESI-MS m/z 725.5[ M+H ]] ⁺ 。

Step 8: synthesis of Compound 18

After 18-7 (0.18 g) and anhydrous DCM (3 mL) were added to the reaction flask, BF was added ₃ .Et ₂ O (0.30 mL) was reacted at room temperature for 0.5h. After the reaction is finished, cooling in ice water bath for 5min, and dripping saturated Na ₂ CO ₃ (10 mL) was quenched, extracted with DCM/MeOH (10:1, 20 mL), the organic phase was evaporated to dryness under reduced pressure and the concentrate was purified by column chromatography (DCM/NH) ₃ in meoh=8:1) to give the target compound 18 (126.0 mg,82.60% yield). ¹ H NMR(500MHz,MeOD)δ7.81(dd,J＝9.1,5.7Hz,1H),7.31–7.22(m,3H),5.48(s,1H),4.96(d,J＝1.5Hz,2H),4.22(dt,J＝10.5,6.8Hz,2H),3.99(s,3H),3.70(d,J＝14.2Hz,1H),3.55(s,3H),3.45(d,J＝13.0Hz,1H),3.34–3.25(m,3H),3.12(dt,J＝10.6,5.5Hz,1H),2.79–2.64(m,2H),2.44(dd,J＝15.8,1.8Hz,1H),2.17–2.07(m,1H),2.01–1.85(m,2H),1.84–1.72(m,4H),1.65(s,1H).ESI-MS m/z:625.3[M+H] ⁺ 。

Example 69 Synthesis of Compound 4- (4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] oct-3-yl) -8-fluoro-5-methoxy-2- (((S) -2-methyltetrahydro-1H-pyrrolizine-7 a (5H) -yl) methoxy) pyrido [4,3-d ] pyrimidin-7-yl) -5-ethynyl-6-fluoronaphthalen-2-ol

Step 1: synthesis of Compound 69-1

M13 (4.84 g,1.86 mmol) was dissolved in anhydrous tetrahydrofuran (85 ml), naH (2.52 g,3.71 mmol) was added, the temperature was reduced to 5 ℃, compound 18-5 (17 g, 1.24 mmol) was dissolved in anhydrous tetrahydrofuran (170 ml), compound 18-5 was slowly added dropwise to the reaction solution, the temperature was controlled to 5-10 ℃, after the dropwise addition, the reaction was continued with stirring at 5-10℃until the starting material was completely reacted. After completion of the reaction, the reaction mixture was quenched with saturated ammonium chloride, diluted with EA (200 ml) and water (200 ml), and the mixture was stirred to give an organic phase, which was extracted once with EA (100 ml), combined with the organic phase, washed once with saturated brine (200 ml), dried over anhydrous sodium sulfate, filtered, dried over a spin-on, and concentrated to give compound 69-1 (19.75 g).

ESI-MS m/z:881.65[M+H] ⁺ 。

Step 2: synthesis of Compound 69-2

Compound 69-1 (19.75 g 22.41 mmol) was dissolved in DMF and CsF (34.05g 224.14mmol) was added and reacted in an oil bath at 35℃for 4h. After the reaction was completed, the temperature was lowered using an ice bath, the temperature was lowered to 10 ℃, water (800 ml) was slowly added dropwise to produce a large amount of solids, and the filter cake was obtained by filtration. This was dissolved with EA (200 ml), washed once with saturated brine (100 ml), and the organic phase was separated and purified by column chromatography (DCM: meoh=20:1) to give compound 69-2 (14.82 g).

ESI-MS m/z:725.51[M+H] ⁺ 。

Step 3: synthesis of Compound 69

Compound 69-2 (14.82 g 20.45 mmol) was dissolved in DCM (140 ml), boron trifluoride etherate (56 ml) was added and reacted in an oil bath at 35℃for 0.5h. After the reaction is completed, the filter cake is obtained by direct filtration. This was dissolved with DCM: meoh=10:1 mixture, the compound was free with saturated sodium carbonate, extracted three times with DCM: meoh=10:1 mixture, washed once with saturated brine, dried over anhydrous sodium sulfate, filtered and spun-dried to give the crude product which was dissolved with methanol, the complex of compound and boron trifluoride was cleaved at 50 ℃, and the cleaved silica gel was purified by column chromatography (DCM: meoh=10:1) to give compound 69 (10.61 g).

ESI-MS m/z:625.47[M+H] ⁺

¹ H NMR(500MHz,DMSO)δ10.16(s,1H),7.97(dd,J＝9.2,5.9Hz,1H),7.47(t,J＝9.0Hz,1H),7.38(d,J＝2.5Hz,1H),7.22(d,J＝2.4Hz,1H),4.90(s,2H),4.08(s,1H),4.02(dd,J＝10.4,4.0Hz,1H),3.98–3.93(m,1H),3.90(s,1H),3.88(s,3H),3.54(d,J＝14.1Hz,1H),3.47(s,2H),3.38(d,J＝12.5Hz,1H),3.32(s,1H),3.18(d,J＝13.9Hz,1H),3.03–2.95(m,1H),2.61–2.52(m,2H),2.35(d,J＝16.2Hz,1H),2.03–1.73(m,4H),1.71-1.64(m,1H),1.63–1.44(m,4H),1.23(s,1H).

Compound 42 was prepared as described in example 1 of reference WO2020216190 A1:

Compound 48 was prepared by the method described in reference WO2022228568 A1; compounds 50-56 are prepared by the method described in reference to WO2022236578A 1.

The examples described below were synthesized using the methods described above, or similar methods using the corresponding intermediates.

TABLE 2

Biological testing

EXAMPLE 1 PK investigation of Compound 7 in combination with the PGY1 inhibitor Elacridar in mice

Compound 7 was mixed with vehicle 10% dmso/10% solutol/80% normal saline, vortexed and sonicated to prepare a clear solution of 1 mg/ml. Balbc female mice of 6 to 7 weeks of age were selected, fasted for 2 hours, orally administered with blank vehicle (20% tween 80/13% ethanol/67% water), orally administered with compound 7 after 15 minutes, whole blood collected for a certain period of time, analyzed for drug concentration by LC-MS/MS method, and drug substitution parameters were calculated using Phoenix WinNolin software (pharside, usa) and the results are shown in table 3.

Table 3 oral compound 7 single drug mice PK data

Route	Compound 7PO
		Dosing Level(mg/kg)	10
Formulation	10% DMSO/10% Solutol/80% normal saline
		Sex	Female
t _1/2 (h)	28
		t _max (h)	2
C _max (ng/mL)	15.7
		AUC _last (h*ng/mL)	173

Compound 7 was mixed with vehicle 10% dmso/10% solutol/80% normal saline, vortexed and sonicated to prepare a clear solution of 1 mg/ml. Elacridar was vortexed with vehicle 20% Tween 80/13% ethanol/67% water, vortexed and sonicated to prepare a clear solution of 3 mg/ml. Balbc female mice of 6 to 7 weeks of age were selected, fasted for 2 hours, orally administered to eladridar, orally administered to compound 7 after 15 minutes, whole blood collected for a certain period of time, analyzed for drug concentration by LC-MS/MS method, and drug generation parameters were calculated using Phoenix WinNolin software (pharside, usa) and the results are shown in table 4.

TABLE 4 oral Compound 7 in combination with Elacridar mouse PK data

Route	Compound 7PO+Elacridar PO
		Dosing Level(mg/kg)	10+30
Formulation	10% DMSO/10% Solutol/80% normal saline
		Sex	Female
t _1/2 (h)	NA
		t _max (h)	7
C _max (ng/mL)	92
		AUC _last (h*ng/mL)	1493

The data in tables 3 and 4 show that the combination of the PGY1 inhibitor eladridar significantly improves the oral PK profile of compound 7 in mice.

EXAMPLE 2 study of the efficacy of Compound 7 in combination with the PGY1 inhibitor Elacridar in Panc04.03 CDX model mice

The experimental method comprises the following steps: construction of a mouse model of subcutaneous xenograft tumor CD-1 of KRAS-G12D mutated human pancreatic cancer cells Panc04.03, 1X 10 ⁷ Each CD-1 nude mouse was inoculated subcutaneously on the right back with Panc04.03 cells (Matrigel mixed with cell suspension at 1:1 volume). When the average tumor volume reaches 200-300mm ³ Within the range, the group administration was started, and 6 tumor-bearing mice were each group (the initial average tumor volume of each group was 229mm ³ )。

The first group G1 was a Vehicle control group (PEG 300: SOLUTOL: citrate buffer=2:1:7).

The second group G2 is compound 7 single drug.

The third group G3 is Compound 7 in combination with Elacridar (day 17-33).

The dosing and experimental protocols are shown in table 5 below.

TABLE 5

Note that: IP represents intraperitoneal injection, PO represents oral gavage, bid represents twice daily.

Tumor size and mouse body weight were measured 2 times per week during the experiment, with each administration referencing the day animal body weight.

The tumor size measurement method comprises the following steps: the length (a) and width (b) of the Tumor were measured using a vernier caliper, and the calculation formula of the Tumor Volume (TV) was: tv=a×b ² /2

TGI% = [ (1- (mean tumor volume at the end of dosing of a treatment group-mean tumor volume at the beginning of dosing of a treatment group))/(mean tumor volume at the end of treatment of a solvent control group-mean tumor volume at the beginning of treatment of a solvent control group) ]x100%.

The results indicated that the second group of G2 compounds 7 had a TGI of 48% after 33 days of 150mg/kg, PO, bid. The third group of G3 compounds 7 had substantially no effect on tumor growth inhibition at 90mg/kg PO, bid after 16 days of administration, and compound 7 (150 mg/kg PO, bid) in combination with eladridar (30 mg/kg PO, QD) was changed from day 17 to effectively inhibit tumor growth, tumor volume was reduced, and TGI at day 33 was 71%, detailed results are shown in FIG. 1. The results show that the combination of the PGY1 inhibitor Elacridar can significantly improve the in vivo oral drug effect of the compound 7 on the KRAS-G12D mutant tumor model.

EXAMPLE 3 PK investigation of Compound 7 in combination with the PGY1 inhibitor Elacridar in Panc04.03 CDX model mice

The experimental method comprises the following steps: whole blood drug concentrations were analyzed by LC-MS/MS method using Panc04.03 mice subjected to animal efficacy study in example 2, and after the last dose on day 33, whole blood (0-0.5-1-2-4-7-10-24 h) was collected for a certain period of time, and drug generation parameters were calculated using Phoenix Winnolin software (Pharsight, USA). And collecting the tumor tissue of the mice after 24 hours of last administration, and measuring the concentration of the compound in the tumor tissue.

Experimental results: c in Whole blood after combination of third group of G3 Compounds 7 with Elacridar compared to the second group of G2 Compounds 7 alone _max ，AUC _last F (F) _last % are significantly improved. The compound concentration in the tumor was also significantly increased, and the results are shown in table 6.

TABLE 6 drug efficacy last dose PK investigation results for Panc04.03CDX model mice

Group of	G2	G3
			Test article	Compound 7	Compound 7+Elacridar
Last dose and mode of administration	150mpk	150mpk+30mpk
			t _1/2 (h)	29.6	20.8
t _max (h)	2.00	4.00
			C _max (ng/mL)	777	2887
AUC _last (h*ng/mL)	10006	50810
			F _last (％)	2.2	11.4
Concentration of Compounds in tumor (ng/g)	6015	54300

The results show that: the combination of the PGY1 inhibitor eladridar significantly increases PK properties in whole blood of panc04.03cdx model mice following oral administration of compound 7, and tumor exposure.

EXAMPLE 4 PK investigation of Compound 1 in combination with the PGY1 inhibitor Elacridar in rats

The experimental method comprises the following steps:

intravenous administration investigation experimental procedure: compound 1 was mixed with vehicle 10% dmso/5% solutol/85% normal saline, vortexed and sonicated to prepare a clear solution of 0.5 mg/ml. SD male rats of 6-7 weeks of age were selected, and after 8 hours of fasted, compound 1 was administered by intravenous injection, whole blood was collected for a certain period of time, the drug concentration was analyzed by the LC-MS/MS method, and the drug substitution parameters were calculated by using Phoenix WinNolin software (Pharsight Co., USA) and the results are shown in Table 7.

TABLE 7 intravenous Compound 1 rat PK data

Oral administration investigation experimental procedure:

compound 1 was mixed with vehicle 10% dmso/5% solutol/85% water, vortexed and sonicated to prepare a clear solution of 1.5 mg/ml. SD male rats of 6-7 weeks of age were selected, and after 8 hours of fasted, compound 1 was orally administered, whole blood was collected for a certain period of time, the drug concentration was analyzed by the LC-MS/MS method, and the drug generation parameters were calculated by using Phoenix WinNolin software (Pharsight Co., USA) and the results are shown in Table 8.

Table 8 oral gavage compound 1 rat PK data

Route	Compound 1PO
		Dosing Level(mg/kg)	30
Formulation	10% DMSO/5% Solutol/85% water
		Sex	male
t _1/2 (h)	NA
		t _max (h)	7
C _max (ng/mL)	3.8
		AUC _last (h*ng/mL)	18
F _last (％)	0.12

Compound 1 was mixed with vehicle 10% dmso/5% solutol/85% water, vortexed and sonicated to prepare a clear solution of 1.5 mg/ml. Elacridar was vortexed with vehicle 20% Tween 80/13% ethanol/67% water, vortexed and sonicated to prepare a clear solution of 3 mg/ml. SD male rats of 6-7 weeks of age were selected, and after 8 hours of fasting, elacridar was orally administered, and after 15 minutes compound 1 was orally administered, whole blood was collected for a certain period of time, and the drug concentration was analyzed by LC-MS/MS method, and the drug generation parameters were calculated by Phoenix Winnolin software (Pharsight Co., U.S.A.), and the results are shown in Table 9.

TABLE 9 oral gavage compound 1+Elacridar rat PK data

Route	Compound 1PO+Elacridar PO
		Dosing Level(mg/kg)	30+30
Formulation	10% DMSO/5% Solutol/85% water
		Sex	male
t _1/2 (h)	7.2809
		t _max (h)	4
C _max (ng/mL)	151
		AUC _last (h*ng/mL)	1831
F _last (％)	12.0

From the results in tables 7-9, it can be seen that the combination of the PGY1 inhibitor eladridar significantly improves the oral PK profile of compound 1 in rats.

Example 5 PK investigation of compound 1 in combination with Elacridar, elbasvir, quinine, diyridamole and encquidar in rats

Compound 1 single drug rat PK study experiment: compound 1 was mixed with vehicle 10% dmso/5% solutol/85% water, vortexed and sonicated to prepare a clear solution of 3 mg/ml. SD male rats of 6 to 7 weeks of age were selected, fasted overnight, orally administered with blank vehicle (20% DMSO/10% Twenn80/70% (20% HP. Beta. CD)), orally administered with compound 1 (G1 group) after 15min, whole blood collected for a certain period of time, analyzed for drug concentration by LC-MS/MS method, and drug substitution parameters were calculated using Phoenix Winnolin software (Pharsight Co., USA).

Compound 1 rat PK study experiment in combination with 5 compounds: compound 1 was mixed with vehicle 10% dmso/5% solutol/85% water, vortexed and sonicated to prepare a clear solution of 3 mg/ml. Elacridar and vehicle 20% DMSO/10% Twen80/70% (20% HP. Beta. CD), vortexing and sonicating to prepare 1.5mg/ml clear solution; elbasvir and vehicle 10% dmso/5% hs15/85% water, vortexed and sonicated to prepare 0.5mg/ml clear solution; quinine and solvent 10% DMSO/5% HS15/85% water are vortexed and sonicated to prepare 9.4mg/ml clear solution; dipyridamole was vortexed with vehicle 10% DMSO/5% HS15/85% water and sonicated to prepare a clear solution of 2.1 mg/ml; encequidar was vortexed with vehicle 20% DMSO/10% HS15/70% (20% HP. Beta. CD), and sonicated to prepare a clear solution of 1.5 mg/ml. SD male rats of 6 to 7 weeks of age were selected, fasted overnight, orally administered with Elacridar (group G2), elbasvir (group G3), quinine (group G4), diyridamole (group G5) and Encequidar (group G6), respectively, orally administered with Compound 1 after 15 minutes, whole blood was collected for a certain period of time, drug concentration was analyzed by LC-MS/MS method, and drug generation parameters were calculated by Phoenix Winnolin software (Pharsight Co., U.S.A.), and the results are shown in Table 10.

Table 10

The results show that: the oral exposure of compound 1 in rats was increased by about 40-fold in combination with eladridar; the oral exposure of compound 1 in rats was increased by about 12-fold in combination with Elbasvir; the combination of Quinine increased the oral exposure of compound 1 in rats by about 4-fold; the oral exposure of compound 1 in rats was increased by about 31-fold in combination with Dipyridamole; the combination encquidar increased the oral exposure of compound 1 in rats by about 11-fold.

EXAMPLE 6 PK investigation of Compound 18 in combination with tetrandrine, elacridar, elbasvir, quinine, diyridamole and Encequidar in mice

Compound 18 single drug mice PK study experiment: compound 18 was mixed with vehicle 10% dmso/5% solutol/85% water, vortexed and sonicated to prepare a clear solution of 1 mg/ml. Balbc female mice of 6 to 7 weeks of age were selected, given blank vehicle (20% dmso/10% tween 80/70% (20% hp βcd)) orally, compound 18 (G1, G4, G7 groups, 3 separate studies) orally after 15min, whole blood was collected for a certain period of time, drug concentrations were analyzed by LC-MS/MS method, and drug generation parameters were calculated using Phoenix WinNolin software (pharshht, usa).

Compound 18 mice PK study experiment in combination with 7 compounds: compound 18 was mixed with vehicle 10% dmso/5% solutol/85% water, vortexed and sonicated to prepare a clear solution of 1 mg/ml. The Elacridar and the menstruum 10 percent propylene glycol/5 percent HS15/85 percent glucose are vortexed and are sonicated to prepare 1.5mg/ml clarified solution; elbasvir and vehicle 10% dmso/5% hs15/85% water, vortexed and sonicated to prepare 1mg/ml clear solution; quinine and solvent 10% DMSO/5% HS15/85% water are vortexed and sonicated to prepare a clear solution of 18.8 mg/ml; dipyridamole was vortexed with vehicle 10% DMSO/5% HS15/85% water and sonicated to prepare a clear solution of 6 mg/ml; encequidar was vortexed with vehicle 20% DMSO/10% HS15/70% (20% HP. Beta. CD), and sonicated to prepare a clear solution of 3 mg/ml. Tetrandrine and vehicle 10% dmso/5% hs15/85% water were vortexed and sonicated to prepare a clear solution of 3 mg/ml. The tetrandrine and the solvent 10% DMSO/5% HS15/85% water are vortexed and sonicated to prepare a clear solution of 3 mg/ml. Balbc female mice of 6 to 7 weeks of age were selected, and tetrandrine (G2), tetrandrine (G3), elacridar (G5), diyridamole (G6), elbasvir (G8), quinine (G9), and Encequidar (G10) were orally administered, respectively, and compound 18 was orally administered after 15min, whole blood was collected for a certain period of time, and drug concentrations were analyzed by LC-MS/MS method, and drug generation parameters were calculated using Phoenix Winnolin software (Pharsight, USA) and the results are shown in Table 11.

TABLE 11

The results show that: the oral exposure of compound 18 in mice was increased by about 2.06-fold in combination with tetrandrine; the oral exposure of compound 18 in mice was increased by about 2.56-fold in combination with tetrandrine; the oral exposure of compound 18 in mice was increased by about 6.24-fold in combination with eladridar; the oral exposure of compound 18 in mice was increased by a factor of about 1.57 in combination with Dipyridamole; the oral exposure of compound 18 in mice was increased by about 2.72-fold in combination with Elbasvir; the oral exposure of compound 18 in mice was increased by about 1.78-fold in combination with Quinine; the combination encquidar increased the oral exposure of compound 18 in mice by about 6.21-fold.

EXAMPLE 7 PK investigation of Compound 12 in combination with Elacridar, diyridamole in mice

Compound 12 single drug mice PK study experiment: compound 12 was mixed with vehicle 10% dmso/5% solutol/85% water, vortexed and sonicated to prepare a clear solution of 1 mg/ml. Balbc female mice of 6 to 7 weeks of age were selected, a blank vehicle 10% dmso/5% hs15/85% water was orally administered, compound 12 (G1 group) was orally administered after 15min, whole blood was collected for a certain period of time, drug concentrations were analyzed by LC-MS/MS method, and drug generation parameters were calculated using Phoenix WinNolin software (pharside, usa).

Compound 12 mice PK study experiments in combination with 2 compounds: compound 12 was mixed with vehicle 10% dmso/5% solutol/85% water, vortexed and sonicated to prepare a clear solution of 1 mg/ml. The Elacridar and the menstruum 10 percent propylene glycol/5 percent HS15/85 percent glucose are vortexed and are sonicated to prepare 1.5mg/ml clarified solution; dipyridamole was vortexed with vehicle 10% DMSO/5% HS15/85% water and sonicated to prepare a clear solution of 6 mg/ml. Balbc female mice of 6 to 7 weeks of age were selected, and eladridar (group G2) and dyridamole (group G3) were orally administered, respectively, and compound 12 was orally administered after 15 minutes, whole blood was collected for a certain period of time, the drug concentration was analyzed by LC-MS/MS method, and the drug substitution parameters were calculated by Phoenix WinNolin software (Pharsight, usa) and the results are shown in table 12.

Table 12

The results show that: the oral exposure of compound 12 in mice was increased by about 8.95-fold in combination with eladridar; the oral exposure of compound 12 in mice was increased by a factor of about 2.51 in combination with Dipyridamole.

EXAMPLE 8 study of the efficacy of Compound 18 in combination with the PGY1 inhibitor Elacridar in Panc04.03 CDX model mice

The experimental method comprises the following steps: construction of KRAS-G12D mutant human pancreatic cancer cell Panc04.03 subcutaneous xenogeneics The CD-1 mouse model of the transplanted tumor is 1×10 ⁷ Each CD-1 nude mouse was inoculated subcutaneously on the right back with Panc04.03 cells (Matrigel mixed with cell suspension at 1:1 volume). When the average tumor volume reaches 200-300mm ³ The administration of groups was started at this time, and 6 tumor-bearing mice per group (the initial mean tumor volume of mice per group was 233-235mm ³ )。

The second group G2 is compound 18 (100 mpk) single drug.

The third group, G3, is compound 18 (10 mpk) in combination with Elacridar.

The fourth group, G4, is compound 18 (30 mpk) in combination with Elacridar.

The dosing and experimental protocols are shown in table 13 below.

TABLE 13

The tumor size measurement method comprises the following steps: the length (a) and width (b) of the Tumor were measured using a vernier caliper, and the calculation formula of the Tumor Volume (TV) was: tv=a×b ² /2；

The TGI results 21 days after administration are shown in table 14.

TABLE 14

The results showed that group G2 compound 18 had a TGI of 104% after 21 days of administration of 100mg/kg PO, bid alone. TGI was 112% after 21 days of G3 group compound 18 (10 mg/kg PO, bid) in combination with Elacridar (30 mpk, PO, bid). Group G4 compound 18 (30 mg/kg PO, bid) was administered in combination with Elacridar (30 mpk, PO, bid) for 21 days with a TGI of 137%. The higher tumor suppression effect was achieved in combination with eladridar at lower doses of compound 18 in the G3 and G4 groups compared to the G2 group alone. The effect of the compound 18 that oral administration is combined with the Elacridar can greatly improve the oral drug effect is embodied.

EXAMPLE 9 PK investigation of Compound 18 in combination with the PGY1 inhibitor Elacridar in Panc04.03 CDX model mice

The experimental method comprises the following steps: whole blood drug concentrations were analyzed by LC-MS/MS method using Panc04.03 mice subjected to animal efficacy study in example 8, and after the last administration on day 21, whole blood was collected for a certain period of time (0-0.5-1-2-4-7-24 h) and drug generation parameters were calculated using Phoenix WinNolin software (Pharsight, USA). And collecting the tumor tissue of the mice after 24 hours of last administration, and measuring the concentration of the compound in the tumor tissue.

Experimental results: c in Whole blood after combination of lower dose (30 mpk) of Compound 18 from group G4 with Elacridar than from group G2 Compound 18 alone (100 mpk) _max ，AUC _last Are significantly higher, the compound concentration in the tumor is also significantly higher, and the results are shown in table 15.

Table 15Panc04.03 CDX model mice and animals drug efficacy last dose PK study results

The results show that: oral combination with eladridar significantly improved PK properties in whole blood and tumor concentrations of compound 18 orally administered in panc04.03 CDX model mice.

Example 10 evaluation of potency of Compounds 18 and 42 as PGY1 substrates Using MDCK-MDR1 cells as a model

The experimental method comprises the following steps: verapamil was used as PGY1 inhibitor at an incubation concentration of 5 μm for compound 18 and compound 42. The rate of transport from tip to base was determined: the top side was added with a transport buffer solution (MEM containing 0.5% BSA) containing compound 18 and compound 42, and the basal side was added with a transport buffer solution; the rate of transport from substrate end to tip was determined: adding a transport buffer solution containing a compound 18 and a compound 42 to the substrate end side, and adding a transport buffer solution to the top end side; for the inhibitor adding group, the inhibitor Verapamil needs to be added to both the delivery buffer at the administration end and the delivery buffer at the receiving end in the MDCK-MDR1 monolayer cell plate. After incubation of the cell plates for 2 hours at 37 ℃, the cell plates were removed and samples were obtained on both sides. LC-MS/MS semi-quantitation was used to obtain compound 18 and compound 42 peak areas. And calculating the bi-directional apparent permeability coefficient of the compound on the cell monolayer membrane according to the peak areas of the compound at the administration end and the receiving end, thereby calculating the efflux rate. The entire experiment was performed in double parallel.

And (3) data processing: data calculations were all performed using Excel, and apparent permeability coefficients (Papp, units: cm/s.times.10) of the compounds were calculated from specific peak areas at the receiving end and at the administration end ^-6 ) Calculated using the following formula:

apparent permeability coefficient (Papp) = (receiving side volume x receiving side drug to internal standard area ratio at end of culture)/(membrane area x culture time x dosing side drug to internal standard area ratio at start of culture x dilution)

Note that: membrane area = 0.33cm ² Recipient side volume=0.7 mL (a-to-B) or 0.2mL (B-to-a), incubation time=7200 seconds;

efflux ratio = Papp (B-ase:Sub>A)/Papp (ase:Sub>A-B);

% recovery = 100× (dosing side drug to internal standard area ratio at end of incubation x dosing side volume + receiving side drug to internal standard area ratio at end of incubation x receiving side volume)/(total dosing side incubation zero minutes)

Substrate determination criteria: it was judged that the efflux ratio of PGY1 substrate was >2 and that the efflux ratio in MDCK-MDR1 cells was reduced by 50% after addition of the inhibitor verapamil.

Experimental results: the permeation results of compound 18 and compound 42 in MDCK-MDR1 cell lines are shown in table 16.

Table 16

Conclusion of experiment: in the absence of verapamil, the average apparent permeability coefficient of Compound 18 in the A-B direction at 5. Mu.M administration concentration was 0.09X 10 ^-6 cm/s; an average apparent permeability coefficient in the B-A direction of 37.5X10 ^-6 cm/s. The discharge ratio was 433.9. Compound 18 showed efflux at 5 μm.

When 5. Mu.M of Compound 18 was incubated with 100. Mu.M verapamil, the average apparent permeability coefficient in the A-B direction was 0.76X10 ^-6 cm/s; an average apparent permeability coefficient in the B-A direction of 4.2X10 ^-6 cm/s. The efflux ratio was 5.5. Compound 18 of the inhibitor group had a greater than 50% decrease in efflux rate at 5 μm.

In conclusion, when verapamil as an inhibitor was not added, the compound 18 concentration was 5. Mu.M, and low permeability was exhibited in the A-B direction. From the data, compound 18 is a PGY1 substrate at a concentration of 5 μm.

In the absence of verapamil, the average apparent permeability coefficient of Compound 42 in the A-B direction was 0.0243 ×10 at a drug administration concentration of 20. Mu.M ^-6 cm/s; an average apparent permeability coefficient in the B-A direction of 6.00×10 ^-6 cm/s. The discharge ratio was 247. Compound 42 showed efflux at 20 μm.

When 20. Mu.M of Compound 42 was incubated with 100. Mu.M verapamil, the average apparent permeability coefficient in the A-B direction was 0.253X 10 ^-6 cm/s; an average apparent permeability coefficient in the B-A direction of 4.29×10 ^-6 cm/s. The discharge ratio was 17.0. Compound 42 of the inhibitor group had a greater than 50% decrease in efflux rate at 20 μm.

In conclusion, when verapamil as an inhibitor was not added, the compound 42 showed low permeability in the a-B direction at a concentration of 20 μm. From the data, compound 42 is a PGY1 substrate at a concentration of 20 μm.

Claims

1. A pharmaceutical combination comprising: (i) KRAS inhibitors; and (ii) a PGY1 inhibitor.

2. The pharmaceutical combination according to claim 1, wherein the KRAS inhibitor is selected from the group consisting of compounds of the following general formula (I), or stereoisomers, tautomers, deuterates or pharmaceutically acceptable salts thereof:

wherein,

l is selected from the group consisting of bond, -cycloalkyl (preferably C) _3-14 Cycloalkyl) -, C _0-3 Alkylene-, -C _0-3 Alkylene-cycloalkyl (preferably C _3-14 Cycloalkyl) -or-C _0-3 Alkylene-cycloalkyl (preferably C _3-14 Cycloalkyl) -C _0-3 An alkylene group; wherein said-cycloalkyl (preferably C _3-14 Cycloalkyl) -, C _0-3 Alkylene-, -C _0-3 Alkylene-cycloalkyl (preferably C _3-14 Cycloalkyl) -, C _0-3 Alkylene-cycloalkyl (preferably C _3-14 Cycloalkyl) -C _0-3 Alkylene-optionally further substituted with one or more R _a Substituted; and is also provided with

When two R _a When the same carbon atom is substituted, two R _a Together with the carbon atoms to which they are attached may form C _3-14 Cycloalkyl, 3-14 membered heterocyclyl;

R ₁ selected from H, -C _0-3 alkylene-N (R) _a ) ₂ 、-C _0-3 Alkylene-heterocyclyl (preferably 3-14 membered heterocyclyl), -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 alkylene-C _6-18 Aryl or-C _0-3 Alkylene-5-18 membered heteroaryl; wherein the-C _0-3 alkylene-N (R) _a ) ₂ 、-C _0-3 Alkylene-heterocyclyl (preferably 3-14 membered heterocyclyl), -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 alkylene-C _6-18 Aryl, -C _0-3 Alkylene-5-18 membered heteroaryl optionally further substituted with one or more R _a Substituted; and is also provided with

When two R _a When the same atom is substituted, two R _a Together with the atoms to which they are attached may form C _3-14 Cycloalkyl, 3-14 membered heterocyclyl;

d is selected from CR ₅ 、C(R ₅ ) ₂ O, N or NR ₆ ；

E is selected from C, CH or N;

f is selected from C, CH, O or N;

independently selected from single bond or double bond;

R ₅ selected from H, hydroxy, oxo, halogen, cyano, C _1-6 Alkyl, C _1-6 Haloalkyl, -S-C _1-6 Alkyl, -C _0-3 alkylene-C _2-4 Alkenyl, C _1-6 Alkoxy, C _1-6 Haloalkoxy or C _3-14 Cycloalkyl;

R ₆ selected from H, hydroxy, halogen, cyano, C _1-6 Alkyl, C _1-6 Haloalkyl, -S-C _1-6 Alkyl, -C _0-3 alkylene-C _2-4 Alkenyl, C _1-6 Alkoxy, C _1-6 Haloalkoxy or C _3-14 Cycloalkyl;

3. The pharmaceutical combination according to claim 2, wherein K in formula (I) is 6-10 membered N-containing

A heterocycle wherein the 6-10 membered N-containing heterocycle is optionally substituted with one or more R _a Substituted; preferably, the 6-10 membered N-containing heterocycle is selected fromPreferably, said R _a Selected from H, cyano, halogen, hydroxy, amino, nitro, C _1-6 Alkyl,/->-C _0-3 Alkylene-cyano, C _1-6 Haloalkyl or C _1-6 An alkoxy group.

4. A pharmaceutical combination according to claim 2 or 3, characterized in thatR in the general formula (I) ₁ Selected from the group consisting of

5. The pharmaceutical combination according to any one of claims 2 to 4, wherein X in formula (I) is selected from the group consisting of a bond, -O-, -NH-, -5-14 membered heterocyclyl-or-C _2-4 Alkynyl-.

6. The pharmaceutical combination according to any one of claims 2 to 5, wherein L in formula (I) is selected from the group consisting of a bond, -cyclopropyl-, -C _0-3 Alkylene-, -C _0-3 alkylene-cyclopropyl-or-C _0-3 alkylene-cyclopropyl-C _0-3 Alkylene-.

7. The pharmaceutical combination according to any one of claims 2 to 6, wherein-X-L-R in formula (I) ₁ Selected from the group consisting of

8. The pharmaceutical combination according to any one of claims 2 to 7, wherein R in formula (I) ₂ Selected from the group consisting of

9. The pharmaceutical combination according to any one of claims 2 to 8, wherein R in formula (I) ₃ Selected from hydrogen, halogen, C _1-6 Alkoxy, -O-CH ₂ -CF ₃ -O-cyclopropyl.

10. The pharmaceutical combination according to any one of claims 2 to 9, wherein R in the general formula (I) ₄ Selected from hydrogen, halogen, C _1-6 Haloalkoxy, oxo, C _1-6 Alkoxy, -O-C _0-3 alkylene-C _3-14 Cycloalkyl, -O-C _0-3 Alkylene-3-14 membered heterocyclyl, -O-C _0-3 alkylene-C _6-18 Aryl or-O-C _0-3 Alkylene-5-18 membered heteroaryl; wherein the-O-C _0-3 alkylene-C _3-14 Cycloalkyl, -O-C _0-3 Alkylene-3-14 membered heterocyclyl, -O-C _0-3 alkylene-C _6-18 Aryl or-O-C _0-3 Alkylene-5-18 membered heteroaryl optionally further substituted with one or more R _a Substitution, said R _a Each independently selected from H, halogen, hydroxy, amino, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl or-C _0-3 Alkylene-5-14 membered heteroaryl.

11. The pharmaceutical combination according to any one of claims 2 to 10, wherein R in the general formula (I) ₅ Selected from hydrogen, halogen, C _1-6 alkyl-S-, oxo, -C _0-3 alkylene-C _2-4 Alkenyl, C _3-6 Cycloalkyl or C _1-6 An alkoxy group.

12. The pharmaceutical combination according to claim 2, wherein the KRAS inhibitor is selected from the group consisting of formula I-a,

An I-B or I-C compound, or a stereoisomer, tautomer, deuterate, or pharmaceutically acceptable salt thereof:

R ₁ selected from the group consisting of

R ₂ Selected from the group consisting of

R ₅ selected from H, hydroxy, halogen, cyano, C _1-6 Alkyl, C _1-6 Haloalkyl, -S-C _1-6 Alkyl, -C _0-3 alkylene-C _2-4 Alkenyl, C _1-6 Alkoxy, C _1-6 Haloalkoxy or C _3-14 Cycloalkyl;

R _a selected from hydrogen, cyano, halogen, hydroxy, amino, nitro, C _1-6 Alkyl group, C _2-6 Alkynyl, -C _0-3 Alkylene-cyano, C _1-6 Haloalkyl, C _1-6 Haloalkoxy, C _1-6 Alkoxy, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8Membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl or-C _0-3 Alkylene-5-14 membered heteroaryl;

13. The pharmaceutical combination according to any one of claims 2-12, wherein the KRAS inhibitor is selected from the compounds in table 1, or stereoisomers, tautomers, deuterides or pharmaceutically acceptable salts thereof:

TABLE 1

14. The pharmaceutical combination according to claim 1, wherein the KRAS inhibitor is selected from KRAS G12D inhibitor or KRAS G12C inhibitor.

15. The pharmaceutical combination of claim 14, wherein the KRAS G12C inhibitor is selected from Sotorasib, adagrasib, GF-105, JDQ-443, YL-15293, D-1553, JAB-21822, ZG-19018, JMKX-001899, HS-10370, GDC-6036, BPI-421286, GH-35, RMC-6291, GEC-255, LY-3537982, MK-1084, D3S-001, HBI-2438, BI-1823911, JS-116, XNW-14010, ABSK-071, ARS-1620, APG-1842, RM-007, erat-3490, MRTX-1257, erat-3691, WDB-178, ICP-915, AZ-8037, ASP-2453, AZD-4625, AU-10458, AU-8653, ATG-012, LC-2, ARS-853, KP-14, RM-018, YF-135, yj-3720, jn-36, or j-20.

16. The pharmaceutical combination according to claim 14, wherein the KRAS G12D inhibitor is selected from MRTX-1133, HRS-4642, JAB-22000, erat-4057, JR-6000, RMC-9805, IMC-KRAS-G12D, KAL-21404358, VRTX-144, RM-036 or KD-8.

17. The pharmaceutical combination according to any one of claims 1-16, wherein the PGY1 inhibitor is selected from Losartan, dactinomycin, levofloxacin, gamidin D, grepafloxacin, rifamycin, lamotrigine, posaconazole, cetirizine, rantidine, loratadine, ranolazine, mefloquine, quinine, chloroquine, dipyridamole, ticagrelor, acetaminophen, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, velplatasvir, voxilaprevir, pibrentasvir, glecaprevir, letermovir, tenofovir, disoproxil, mirabegron, bisoprolol, verapamil, mibefradil, nicardipine, amlodipine, diltiazem, tezacaftor, elexacaftor, taurocholic acid, bromocriptine, chlorpromazine, haloperidol, paliperidone, elagolix, enasidenib, lvosidenib, cyclosporine, erythromycin, clarithromycin, azithromycin, indomethacin, slidenafil, vardenafil, valinomycin, esomeprazole, pantoprazole, omeprazole, lansoprazole, lasmiditan, canagliflozin, venlafaxine, quinidine, citalopram, prazosin, atorvastain, methyl blue, tolvaptan, encequidar, dexverapamil, dexniguldipine, elacridar, tariquidar (XR-9576), zosuquidar, mitotane, laniquidar, valspodar, tezacaftor, piperine, timcodar dimesilate, licochalcone a, dofequidar fumarate, biricodar, cinchonine, cyclosporine (hoons), CTP-786, STI-0529, BST-204, schisandrin B, KI-1102, CBT-1, ORX-102, ORX-101, ATNX-04, WS-10, CAP-0121, WS-691, WS-898, YS-370, TTT-28, CP-778875, EMD 55900, PMI-002, HE-10, SDZ-280-446, W-198, BIBW-22, S-9788, ONT-093, CRL-1336, z-280125, LYP-10426.

18. The pharmaceutical combination according to claim 17, wherein the PGY1 inhibitor is selected from Amlodipine, verapamil, bisoprolol, nicardipine, diltiazem, ticagrelor, dipyridamole, prazosin, quinidine, atorvastatin, canagliflozin, cyclosporine, indomethacin, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, glecaprevir, tenofovir disoproxil, quinine, chloroquine, dexverapamil, dexniguldipine, valspodar, dofequidar fumarate, zosuquidar, laniquidar, mitotane, biricodar, elacridar, ONT-093, encequidar, tezacaftor or Piperine.

19. The pharmaceutical combination according to any one of claims 1 to 16, the PGY1 inhibitor is selected from cyclosporin A cycloporine A, progesterone Progesterone, propafenone Propanone, nifedipine, felodipine felopirine, aureobasidin A, fluphenazine flufenazine, chlorpromazine chloromazine, caffeine, yohimbine, reserpine, ganciclovibrolamine, tamoxifen, tolrimitinib, ritonafen, tacrolimus Tacrolimus, zolidazoquidambarir, ranolazine, itraconazole, tioconazole, tiatavanavir, saquinavir; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

20. The pharmaceutical combination according to claim 1, wherein the KRAS inhibitor is selected from formula (I)

The compound, or a stereoisomer, tautomer, deuterate, or pharmaceutically acceptable salt thereof:

wherein,

d is selected from CR ₅ 、C(R ₅ ) ₂ O, N or NR ₆ ；

E is selected from C, CH or N;

f is selected from C, CH, O or N;

independently selected from single bond or double bond;

R ₄ selected from the group consisting of absent, H, halogen, oxo,C _1-6 Alkoxy, C _1-6 Haloalkoxy, -O-C _0-3 alkylene-C _3-14 Cycloalkyl, -O-C _0-3 Alkylene-3-14 membered heterocyclyl, -O-C _0-3 alkylene-C _6-18 Aryl or-O-C _0-3 Alkylene-5-18 membered heteroaryl; wherein the-O-C _0-3 alkylene-C _3-14 Cycloalkyl, -O-C _0-3 Alkylene-3-14 membered heterocyclyl, -O-C _0-3 alkylene-C _6-18 Aryl or-O-C _0-3 Alkylene-5-18 membered heteroaryl optionally further substituted with one or more R _a Substitution;

R _a each independently selected from H, halogen, hydroxy, amino, oxo, nitro, cyano, carboxyl, C _2-6 Alkynyl, C _1-6 Alkyl, C _2-6 Olefins, -C _1-3 alkylene-C _2-6 Olefins, C _1-6 Hydroxyalkyl, C _1-6 Aminoalkyl, -C _1-3 Alkylene-cyano, C _1-6 Haloalkyl, -C _0-3 alkylene-C _1-6 Alkoxy, C _1-6 alkyl-S-, C _1-6 Haloalkoxy, C _1-6 Heteroalkyl, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 Alkylene group-C _6-14 Aryl or-C _0-3 Alkylene-5-14 membered heteroaryl; wherein the C _1-6 Alkyl, C _2-6 Olefins, -C _1-3 alkylene-C _2-6 Olefins, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl, -C _0-3 Alkylene-5-14 membered heteroaryl optionally further substituted with one or more halo, hydroxy, cyano, hydroxy, amino, nitro, C _1-6 Alkyl, C _3-14 Cycloalkyl, 3-8 membered heterocyclyl, C _6-14 Aryl or 5-14 membered heteroaryl;

the PGY1 inhibitor is selected from Amlodipine, verapamil, bisoprolol, nicardipine, diltiazem, ticagrelor, dipyridamole, prazosin, quinidine, atorvastatin, canagliflozin, cyclosporine, indomethacin, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, glecaprevir, tenofovir disoproxil, quinine, chloroquine, dexverapamil, dexniguldipine, valspodar, dofequidar fumarate, zosuquidar, laniquidar, mitotane, biricodar, elacridar, ONT-093, encequidar, tezacaftor, piperine, cyclosporin A cycloporine A, progesterone Progesterone, propafenone, nifedipine, felodipine, aureobasidin A, fluphenazine flufenazine, chlorpromazine, caffeine, yohimbine, reserpine, ganamazolamide, tamoxifen, tolrimifene, dariumine Darotamamide, futanium Fomaminib, ritonavir, tacrolimus, oxazolquidamid Zosuquivir, razinol, itraconazole, tiiramate; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

21. The pharmaceutical combination according to claim 1, wherein the KRAS inhibitor is selected from the group consisting of formulas I-a

R ₁ selected from the group consisting of

R ₂ Selected from the group consisting of

R _a selected from hydrogen, cyano, halogen, hydroxy, amino, nitro, C _1-6 Alkyl group, C _2-6 Alkynyl, -C _0-3 Alkylene-cyano, C _1-6 Haloalkyl, C _1-6 Haloalkoxy, C _1-6 Alkoxy, -C _0-3 alkylene-C _3-14 Cycloalkyl, -C _0-3 Alkylene-3-8 membered heterocyclyl, -C _0-3 alkylene-C _6-14 Aryl or-C _0-3 Alkylene groupA 5-to 14-membered heteroaryl;

22. The pharmaceutical combination of claim 1, wherein the KRAS inhibitor is selected from a compound of formula I-B, or a stereoisomer, tautomer, deuterate, or pharmaceutically acceptable salt thereof:

R ₁ selected from the group consisting of

R ₂ Selected from the group consisting of

23. Pharmaceutical combination according to claim 1, characterized in that the KRAS inhibitor is selected from compounds of formula I-C, or stereoisomers, tautomers, deuterides or pharmaceutically acceptable salts thereof:

R ₁ selected from the group consisting of

R ₂ Selected from the group consisting of

the PGY1 inhibitor is selected from Amlodipine, verapamil, bisoprolol, nicardipine, diltiazem, ticagrelor, dipyridamole, prazosin, quinidine, atorvastatin, canagliflozin, cyclosporine, indomethacin, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, glecaprevir, tenofovir disoproxil, quinine, chloroquine, dexverapamil, dexniguldipine, valspodar, dofequidar fumarate, zosuquidar, laniquidar, mitotane, biricodar, elacridar, ONT-093, encequidar, tezacaftor or Piperine, cyclosporin A cycloporine A, progesterone Progesterone, propafenone Propanone, nifedipine, felodipine, aureobasidin A, fluphenazine flufenazine, chlorpromazine, caffeine, yohimbine reserpine, ganciclovir, tamoxifen, toremifene Dalutamide, futamitinib Fostamatinib, ritonavir Tacrolimus, zoquidambar Zosuquidar, ranolazine, itraconazole Iraconazole, telanavir Tiprataavir, saquinavir; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

24. The pharmaceutical combination according to claim 1, wherein the KRAS inhibitor is selected from the compounds of table 1, or stereoisomers, tautomers, deuterides or pharmaceutically acceptable salts thereof; the PGY1 inhibitor is selected from Amlodipine, verapamil, bisoprolol, nicardipine, diltiazem, ticagrelor, dipyridamole, prazosin, quinidine, atorvastatin, canagliflozin, cyclosporine, indomethacin, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, glecaprevir, tenofovir disoproxil, quinine, chloroquine, dexverapamil, dexniguldipine, valspodar, dofequidar fumarate, zosuquidar, laniquidar, mitotane, biricodar, elacridar, ONT-093, encequidar, tezacaftor, piperine, cyclosporin A cycloporine A, progesterone Progesterone, propafenone, nifedipine, felodipine, aureobasidin A, fluphenazine flufenazine, chlorpromazine, caffeine, yohimbine, reserpine, ganamazolamide, tamoxifen, tolrimifene, dariumine Darotamamide, futanium Fomaminib, ritonavir, tacrolimus, oxazolquidamid Zosuquivir, razinol, itraconazole, tiiramate; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

25. The pharmaceutical combination according to claim 14, wherein the KRAS G12D inhibitor is selected from MRTX-1133, HRS-4642, JAB-22000, erat-4057, JR-6000, RMC-9805, IMC-KRAS-G12D, KAL-21404358, VRTX-144, RM-036 or KD-8; the PGY1 inhibitor is selected from Amlodipine, verapamil, bisoprolol, nicardipine, diltiazem, ticagrelor, dipyridamole, prazosin, quinidine, atorvastatin, canagliflozin, cyclosporine, indomethacin, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, glecaprevir, tenofovir disoproxil, quinine, chloroquine, dexverapamil, dexniguldipine, valspodar, dofequidar fumarate, zosuquidar, laniquidar, mitotane, biricodar, elacridar, ONT-093, encequidar, tezacaftor, piperine, cyclosporin A cycloporine A, progesterone Progesterone, propafenone, nifedipine, felodipine, aureobasidin A, fluphenazine flufenazine, chlorpromazine, caffeine, yohimbine, reserpine, ganamazolamide, tamoxifen, tolrimifene, dariumine Darotamamide, futanium Fomaminib, ritonavir, tacrolimus, oxazolquidamid Zosuquivir, razinol, itraconazole, tiiramate; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

26. The pharmaceutical combination of claim 14, wherein the KRAS G12C inhibitor is selected from Sotorasib, adagrasib, GF-105, JDQ-443, YL-15293, D-1553, JAB-21822, ZG-19018, JMKX-001899, HS-10370, GDC-6036, BPI-421286, GH-35, RMC-6291, GEC-255, LY-3537982, MK-1084, D3S-001, HBI-2438, BI-1823911, JS-116, XNW-14010, ABSK-071, ARS-1620, APG-1842, RM-007, erat-3490, MRTX-1257, erat-3691, WDB-178, ICP-915, AZ-8037, ASP-2453, AZD-4625, AU-10458, AU-8653, ATG-012, LC-2, ARS-853, KP-14, RM-018, YF-135, YF-3720, jn-032, jn-36, or j-20; the PGY1 inhibitor is selected from Amlodipine, verapamil, bisoprolol, nicardipine, diltiazem, ticagrelor, dipyridamole, prazosin, quinidine, atorvastatin, canagliflozin, cyclosporine, indomethacin, asunaprevir, telaprevir, simeprevir, daclatasvir, elbasvir, glecaprevir, tenofovir disoproxil, quinine, chloroquine, dexverapamil, dexniguldipine, valspodar, dofequidar fumarate, zosuquidar, laniquidar, mitotane, biricodar, elacridar, ONT-093, encequidar, tezacaftor, piperine, cyclosporine ACyclosporine A, progesterone Progesterone, propafenone Propafenone, nifedipine, felodipine, aureobasidin A, fluphenazine flufenazine, chlorpromazine, caffeine, yohimbine, reserpine, ganapamizol, tamoxifen, tolnaftate, ritonavir, tacrolimus Tacrolimus, zoquin Zosuquivir, razinol fluvozine, itraconazole, titansvara fluvozole; tarimquidar, tetrandrine or Tetrandrine Demethyl Tetrandrine.

27. The pharmaceutical combination according to any one of claims 1 to 26, wherein the ratio of the average daily dose of KRAS inhibitor to PGY1 inhibitor is selected from 100:1 to 1:100, optionally the ratio of the average daily dose is selected from 20:1 to 1:20, 1:10 to 10:1, 1:5 to 5:1, 1:4 to 4:1, 1:3 to 3:1, 1:2 to 2:1 or 1:1.

28. The pharmaceutical combination of any one of claims 1-27, wherein the KRAS inhibitor is administered three times per day, twice per day, once per two days, once per three days, once per four days, once per five days, once per six days, once per week, once per two weeks, or once every three weeks.

29. The pharmaceutical combination according to any one of claims 1 to 28, wherein the KRAS inhibitor is administered at a dose of 1 to 5000mg per administration; preferably, the KRAS inhibitor is administered at a dose of 1-1000mg, 1-500mg, 5-500mg, 10-500mg, 50-400mg, 50-300mg, 50-200mg, 50-100mg, 100-500mg, 100-400mg, 100-300mg or 100-200mg per time.

30. The pharmaceutical combination of any one of claims 1-29, wherein the PGY1 inhibitor is administered three times per day, twice per day, once per two days, once per three days, once per four days, once per five days, once per six days, once per week, once per two weeks, or once every three weeks.

31. The pharmaceutical combination according to any one of claims 1 to 36, wherein the PGY1 inhibitor is administered at a dose of 0.01-1000mg per administration; preferably, the PGY1 inhibitor is administered at a dose of 0.1-100mg, 0.1-80mg, 0.1-50mg, 0.1-20mg, 1-100mg, 1-50mg, 5-100mg, 5-50mg, 5-30mg, 10-100mg, 10-50mg, or 10-30mg per time.

32. The pharmaceutical combination according to any one of claims 1-37, wherein the pharmaceutical combination is a fixed combination; optionally the fixed combination is in the form of a solid pharmaceutical composition; optionally, the KRAS inhibitor and PGY1 inhibitor in the fixed combination are present in the same solid pharmaceutical composition.

33. The pharmaceutical combination according to any one of claims 1-32, wherein the pharmaceutical composition is a non-fixed combination; optionally, the KRAS inhibitor and PGY1 inhibitor in the non-fixed combination are each in the form of a solid pharmaceutical composition; optionally, each of the KRAS inhibitor and PGY1 inhibitor in the non-fixed combination is in the form of a solid pharmaceutical composition, and the solid pharmaceutical composition of the KRAS inhibitor and the solid pharmaceutical composition of the PGY1 inhibitor are present in the same pouch; optionally, each of the KRAS inhibitor and PGY1 inhibitor in the non-fixed combination is in the form of a solid pharmaceutical composition, and the solid pharmaceutical composition of the KRAS inhibitor and the solid pharmaceutical composition of the PGY1 inhibitor are not present in the same pouch.

34. The pharmaceutical combination according to any one of claims 1-33, wherein the KRAS inhibitor and PGY1 inhibitor can be administered to the patient simultaneously, separately and/or sequentially.

35. The pharmaceutical combination according to any one of claims 1-34, wherein the pharmaceutical combination is an oral formulation.

36. The pharmaceutical combination according to any one of claims 1-35, wherein the pharmaceutical combination is a tablet or capsule.

37. A kit comprising the pharmaceutical combination of any one of claims 1-36.

38. Use of a pharmaceutical combination according to any one of claims 1-36 or a kit according to claim 37 in the manufacture of a medicament for the treatment of KRAS mediated diseases.

39. The use according to claim 38, wherein the KRAS mediated disease is a cancer, preferably selected from breast cancer, multiple myeloma, bladder cancer, endometrial cancer, gastric cancer, cervical cancer, rhabdomyosarcoma, non-small cell lung cancer, polymorphic lung cancer, ovarian cancer, esophageal cancer, melanoma, colorectal cancer, hepatoma, head and neck cancer, hepatobiliary cell cancer, myelodysplastic syndrome, glioblastoma, prostate cancer, thyroid cancer, xu Wangshi cell tumor, lung squamous cell carcinoma, bryoid keratosis, synovial sarcoma, skin cancer, pancreatic cancer, testicular cancer or liposarcoma.

40. A kit for treating a KRAS mediated disease comprising (i) the KRAS inhibitor of any one of claims 1-33, and (ii) the PGY1 inhibitor of any one of claims 1-33, wherein: (i) Contained in a first compartment, and (ii) contained in a second compartment.