CN112778334B

CN112778334B - Aromatic heterocyclic compound, pharmaceutical composition and application thereof

Info

Publication number: CN112778334B
Application number: CN202011230384.8A
Authority: CN
Inventors: 张健存; 邹晴安; 陈延维; 康宁; 张礼军; 胡洋; 游华金; 肖学兵
Original assignee: Guangzhou Henovcom Bioscience Co ltd
Current assignee: Guangzhou Henovcom Bioscience Co ltd
Priority date: 2019-11-07
Filing date: 2020-11-06
Publication date: 2022-04-15
Anticipated expiration: 2040-11-06
Also published as: WO2021088957A1; CN112778334A

Abstract

The present invention provides a novel class of heteroaromatic compounds as atx (autotaxin) inhibitors, pharmaceutical compositions comprising the compounds, and methods of using the compounds and compositions to treat a disease in a mammal having a pathological feature of increased atx (autotaxin) expression, wherein the compounds have a structure according to formula (I) or (II), or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, nitrogen oxide, metabolite, prodrug, or mixture thereof:

wherein R is^1a、R²、R³、R⁶、Cy¹、Cy²、Y¹、Y²Z and t all have the definitions given in the present invention.

Description

Aromatic heterocyclic compound, pharmaceutical composition and application thereof

Technical Field

The invention relates to the field of medicinal chemistry, in particular to a novel aromatic heterocyclic compound serving as an ATX (Autotaxin) inhibitor, a pharmaceutical composition containing the compound and application of the compound or the composition in treating diseases with pathological characteristics of increased expression of ATX (Autotaxin).

Background

Autotaxin (ATX), the first isolated protein from A2058 melanoma cells in 1992, was called an "autocrine motor" and is a secreted glycoprotein. ATX has Phosphodiesterase (PDE) activity and is a member of the extracellular pyrophosphatase/phosphodiesterase (ENPP) family. ATX also has lysophospholipase d (lysopld) activity, and can catalyze the production of lysophosphatidic acid (LPA) using Lysophosphatidylcholine (LPC) as a substrate. LPA is not only a precursor for phospholipid synthesis, but also can cause a wide range of biological effects through a variety of signaling pathways. LPA, once produced, is capable of functioning mediated by six cell surface specific receptor proteins (LPA1-6), the G protein-coupled receptors (GPCRs). According to the nomenclature of endothelial cell differentiation genes (Edg) and ventricular region genes, LPA1-6 is LPA1/Edg-2/VZG-1, LPA2/Edg-4, LPA3/Edg-7, LPA4/p2Y9/GPR23, LPA5/GPR92 and LPA6/p2Y5, respectively, each of which is mediated by a G.alpha.protein (Gs, Gi, Gq, and G12/13) to trigger a series of cell signaling cascades. Wherein the major pathway involves hydrolysis of phosphatidylinositol diphosphate (PIP2), which in turn triggers intracellular calcium ion release and protein kinase c (pkc) activation; inhibition of the adenylate cyclase (cAMP) signaling pathway; activating Ras-MAPK, MERK and ERK channels, and regulating cell proliferation activity; activating phosphoinositide PI3K-AKT pathway, regulating cell survival and apoptosis activity; finally, activation of the Rho pathway regulates cytoskeletal remodeling, shape change, and cell migration activities. In many pathological conditions, especially in tumor cells, ATX is highly expressed, resulting in excessive LPA concentrations. In tumor cells, LPA concentrations can rise to 10. mu. mol/L, well above the normal level of 100 nmol/L. Excessive LPA increases the production of Vascular Endothelial Growth Factor (VEGF), promotes angiogenesis; reduce the expression of the tumor suppressor factor p53, and increase the survival and metastasis of tumor cells. The ATX-LPA signaling pathway is involved in many physiological and pathological processes and thus has important implications for a number of serious diseases, including mainly cardiovascular diseases, autoimmune diseases, cancer, fibrotic diseases, inflammation, neurological diseases, pain, etc. LPA has multiple functions in tumorigenesis, promotes growth, angiogenesis, metastasis and development of drug resistance in tumor cells. Therefore, the concentration level of LPA is reduced, and the treatment and control of tumors are facilitated. Correspondingly, the inhibition of the activity of AXT and the blockage of the generation pathway of LPA are hot spots for the research of treating various serious diseases.

With the ongoing and intensive research on ATX, many new inhibitors targeting it have been promoted, with cancer and fibrotic diseases being the most focused on. Fibrotic diseases are mainly Idiopathic Pulmonary Fibrosis (IPF) and liver fibrosis. IPF is a fatal disease that manifests as diffuse alveolitis and alveolar disorganization and leads to progressive development of pulmonary interstitial fibrosis with a poor prognosis and an average survival time of 2 to 5 years. IPF is probably the most closely linked disease to the ATX-LPA pathway, since in lung tissue, ATX expression is most concentrated in bronchial epithelial cells and alveolar macrophages, which can juxtapose fibroblasts.

Currently, GLPG-1690, as an Autotaxin inhibitor, has entered the clinical phase II trial phase for the treatment of idiopathic pulmonary fibrosis; the concentration of ATX in serum is closely related to the hardness values of liver fibrosis and liver, and is one of the best indexes for predicting liver cirrhosis. In addition, ATX is highly expressed in many tumor tissues, including melanoma, non-small cell lung cancer, liver cancer, kidney cancer, breast cancer, thyroid cancer, ovarian cancer, and hodgkin's lymphoma. LPA/ATX promotes cell invasion and metastasis during tumor cell growth. Therefore, the ATX inhibitor blocks a signal transduction pathway, and provides a new way for clinically treating cancers and fibrotic diseases.

Compared with the traditional kinase inhibitor, the ATX inhibitor has the advantages that the ATX inhibitor can inhibit the activity of ATX and simultaneously affect a plurality of signal paths related to cell proliferation, growth, apoptosis and the like, has a good inhibition effect on some drug-resistant tumors, is closely related to the fibrosis of a plurality of organs, and is an important target for researching and developing novel fibrotic disease drugs.

The invention provides a new aromatic heterocyclic compound which has good inhibitory activity on ATX. The compound has excellent drug effect, pharmacokinetic property and/or toxicological property, and has good clinical application prospect.

Description

Definitions and general terms

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents and publications referred to herein are incorporated by reference in their entirety.

The following definitions as used herein should be applied, unless otherwise indicated. For the purposes of the present invention, the chemical elements are in accordance with the CAS version of the periodic Table of the elements, and the handbook of chemistry and Physics, 75 th edition, 1994. In addition, general principles of Organic Chemistry can be found in the descriptions of "Organic Chemistry", Thomas Sorrell, University Science Books, Sausaltito: 1999, and "March's Advanced Organic Chemistry" by Michael B. Smith and Jerry March, John Wiley & Sons, New York:2007, the entire contents of which are incorporated herein by reference.

The articles "a," "an," and "the" as used herein are intended to include "at least one" or "one or more" unless otherwise indicated or clearly contradicted by context. Thus, as used herein, the articles refer to articles of one or more than one (i.e., at least one) object. For example, "a component" refers to one or more components, i.e., there may be more than one component contemplated for use or use in embodiments of the described embodiments.

"stereoisomers" refers to compounds having the same chemical structure but differing in the arrangement of atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformers (rotamers), geometric isomers (cis/trans), atropisomers, and the like.

The term "tautomer" or "tautomeric form" refers to structural isomers having different energies that can interconvert by a low energy barrier (low energy barrier). If tautomerism is possible (e.g., in solution), then the chemical equilibrium of the tautomer can be reached. For example, proton tautomers (also known as proton transfer tautomers) include interconversions by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers (valenctautomers) include interconversion by recombination of some of the bonding electrons. A specific example of keto-enol tautomerism is the tautomerism of the pentan-2, 4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerism is phenol-ketone tautomerism. One specific example of phenol-ketone tautomerism is the tautomerism of pyridin-4-ol and pyridin-4 (1H) -one tautomers. Unless otherwise indicated, all tautomeric forms of the compounds of the invention are within the scope of the invention.

In general, the term "substituted" means that one or more hydrogen atoms in a given structure are replaced with a particular substituent. Unless otherwise indicated, a substituted group may have one substituent substituted at each substitutable position of the group. When more than one position in a given formula can be substituted with one or more substituents selected from a particular group, the substituents may be substituted at each position, identically or differently.

The term "unsubstituted" means that the specified group bears no substituents.

The term "optionally substituted with … …", is used interchangeably with the term "unsubstituted or substituted with ….," i.e., the structure is unsubstituted or substituted with one or more substituents described herein, including but not limited to, H, D, F, Cl, Br, I, N₃、CN、NO₂、OH、SH、NH₂Oxo (C ═ O), alkyl, haloalkyl, cyano-substituted alkyl, hydroxyalkyl, alkoxy, haloalkoxy, or haloalkoxyalkyl, and the like.

In addition, unless otherwise explicitly indicated, the descriptions of the terms "… independently" and "… independently" and "… independently" used in the present invention are interchangeable and should be understood in a broad sense to mean that the specific items expressed between the same symbols do not affect each other in different groups or that the specific items expressed between the same symbols in the same groups do not affect each other.

In the various parts of this specification, substituents of the disclosed compounds are disclosed in terms of group type or range. It is specifically intended that the invention includes each and every independent subcombination of the various members of these groups and ranges. For example, the term "C_1-6Alkyl "means in particular independently disclosed methyl, ethyl, C₃Alkyl radical, C₄Alkyl radical, C₅Alkyl and C₆An alkyl group.

The term "alkyl" or "alkyl group" as used herein, denotes a saturated, straight or branched chain monovalent hydrocarbon radical containing from 1 to 20 carbon atoms, wherein the alkyl group may be optionally substituted with one or more substituents as described herein. Unless otherwise specified, alkyl groups contain 1-20 carbon atoms. In one embodiment, the alkyl group contains 1 to 12 carbon atoms; in another embodiment, the alkyl group contains 1 to 6 carbon atoms; in yet another embodiment, the alkyl group contains 1 to 4 carbon atoms; in yet another embodiment, the alkyl group contains 1 to 3 carbon atoms. The alkyl group may be optionally substituted with one or more substituents described herein.

Examples of alkyl groups include, but are not limited to, methyl (Me, -CH)₃) Ethyl group (Et, -CH)₂CH₃) N-propyl (n-Pr, -CH)₂CH₂CH₃) Isopropyl group (i-Pr, -CH (CH)₃)₂) N-butyl (n-Bu, -CH)₂CH₂CH₂CH₃) Isobutyl (i-Bu, -CH)₂CH(CH₃)₂) Sec-butyl (s-Bu, -CH (CH)₃)CH₂CH₃) Tert-butyl (t-Bu, -C (CH)₃)₃) N-pentyl (-CH)₂CH₂CH₂CH₂CH₃) 2-pentyl (-CH (CH)₃)CH₂CH₂CH₃) 3-pentyl (-CH (CH)₂CH₃)₂) 2-methyl-2-butyl (-C (CH)₃)₂CH₂CH₃) 3-methyl-2-butyl (-CH (CH)₃)CH(CH₃)₂) 3-methyl-1-butyl (-CH)₂CH₂CH(CH₃)₂) 2-methyl-1-butyl (-CH)₂CH(CH₃)CH₂CH₃) N-hexyl (-CH)₂CH₂CH₂CH₂CH₂CH₃) 2-hexyl (-CH (CH)₃)CH₂CH₂CH₂CH₃) 3-hexyl (-CH (CH)₂CH₃)(CH₂CH₂CH₃) 2-methyl-2-pentyl (-C (CH))₃)₂CH₂CH₂CH₃) 3-methyl-2-pentyl (-CH (CH)₃)CH(CH₃)CH₂CH₃) 4-methyl-2-pentyl (-CH (CH)₃)CH₂CH(CH₃)₂) 3-methyl-3-pentyl (-C (CH)₃)(CH₂CH₃)₂) 2-methyl-3-pentyl (-CH (CH)₂CH₃)CH(CH₃)₂)2, 3-dimethyl-2-butyl (-C (CH)₃)₂CH(CH₃)₂)3, 3-dimethyl-2-butyl (-CH (CH)₃)C(CH₃)₃) N-heptyl, n-octyl, and the like.

The term "alkoxy" means an alkyl group attached to the rest of the molecule through an oxygen atom, wherein the alkyl group has the definition as described herein. Unless otherwise specified, the alkoxy group contains 1 to 12 carbon atoms. In one embodiment, the alkoxy group contains 1 to 6 carbon atoms; in another embodiment, the alkoxy group contains 1 to 4 carbon atoms; in yet another embodiment, the alkoxy group contains 1 to 3 carbon atoms. The alkoxy group may be optionally substituted with one or more substituents described herein.

Examples of alkoxy groups include, but are not limited to, methoxy (MeO, -OCH)₃) Ethoxy (EtO, -OCH)₂CH₃) 1-propoxy (n-PrO, n-propoxy, -OCH)₂CH₂CH₃) 2-propoxy (i-PrO, i-propoxy, -OCH (CH)₃)₂) 1-butoxy (n-BuO, n-butoxy, -OCH)₂CH₂CH₂CH₃) 2-methyl-l-propoxy (i-BuO, i-butoxy, -OCH)₂CH(CH₃)₂) 2-butoxy (s-BuO, s-butoxy, -OCH (CH)₃)CH₂CH₃) 2-methyl-2-propoxy (t-BuO, t-butoxy, -OC (CH)₃)₃) 1-pentyloxy (n-pentyloxy, -OCH)₂CH₂CH₂CH₂CH₃) 2-pentyloxy (-OCH (CH)₃)CH₂CH₂CH₃) 3-pentyloxy (-OCH (CH))₂CH₃)₂) 2-methyl-2-butoxy (-OC (CH))₃)₂CH₂CH₃) 3-methyl-2-butoxy (-OCH (CH)₃)CH(CH₃)₂) 3-methyl-l-butoxy (-OCH)₂CH₂CH(CH₃)₂) 2-methyl-l-butoxy (-OCH)₂CH(CH₃)CH₂CH₃) And so on.

The term "hydroxyalkyl" as used herein means an alkyl group substituted with one or more hydroxyl groups, wherein the alkyl group has the definition as set forth herein, examples of which include, but are not limited to, hydroxyethyl, 2-hydroxypropyl, hydroxymethyl, and the like.

The terms "heterocyclyl" and "heterocycle" are used interchangeably herein and, unless otherwise specified, refer to monovalent monocyclic non-aromatic ring systems and/or polycyclic ring systems comprising at least one non-aromatic ring; wherein one or more (in certain embodiments, 1,2,3, or 4) of said non-aromatic monocyclic atoms is independently selected from O, S (O)_0-2And N, and the remaining ring atoms are carbon atoms; and wherein one or more (in certain embodiments, 1,2,3, or 4) of the ring atoms of the polycyclic ring system is independently selected from O, S (O)_0-2And N, and the remaining ring atoms are all carbon atoms. In some embodiments, the heterocyclic ring contains 1 or 2 heteroatoms, each of which is a nitrogen atom. In some embodiments, the heterocyclic group is polycyclic and contains one heteroatom in a non-aromatic ring, or one heteroatom in an aromatic ring, or two heteroatoms with one in an aromatic ring and the other in a non-aromatic ring. In some embodiments, the heterocyclyl group has 3-20, 3-15, 3-10, 3-8, 4-7, or 5-6 ring atoms. In some embodiments, the heterocyclyl is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system. In some embodiments, the heterocyclyl group may be a bridged or unbridged, spiro or unbridged, and/or fused or unfused bicyclic group. One or more nitrogen and sulfur atoms may optionally be oxidized, and one or more nitrogen atoms mayOptionally quaternized, one or more carbon atoms optionally being substituted

And (6) replacing. Some rings may be partially or fully saturated or aromatic, provided that the heterocyclic ring is not fully aromatic. The monocyclic heterocycle and polycyclic heterocycle may be attached to the main structure at any heteroatom or carbon atom that results in the creation of a stable compound. The polycyclic heterocyclic group may be attached to the main structure through any ring thereof, including any aromatic or non-aromatic ring, regardless of whether the ring contains heteroatoms. In some embodiments, heterocyclyl is "heterocycloalkyl" which is 1) a saturated or partially unsaturated (but not aromatic) monovalent monocyclic group containing at least one ring heteroatom as described herein, or 2) a saturated or partially unsaturated (but not aromatic) monovalent bicyclic or tricyclic group in which at least one ring contains at least one heteroatom as described herein. When the heterocyclyl and heterocycloalkyl group are substituted, they may be substituted on either ring, i.e., on any aromatic or non-aromatic ring contained by the heterocyclyl and heterocycloalkyl groups. In some embodiments, such heterocyclyl groups include, but are not limited to, oxiranyl, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, 1, 3-dioxocyclopentyl, dithiocyclopentyl, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, dioxanyl, dithianyl, thioxanyl, homopiperazinyl, homopiperidinyl, oxepanyl, thietanyl, oxazepanyl, oxazepinyl, and thiazepinyl

Radical, diaza

Radical, S-N-aza

A group, a benzodioxanyl group, a benzodioxolyl group, a benzofuranonyl group, a benzopyranonyl group, a benzopyranyl group, a dihydrobenzofuranyl group, a benzothiophenyl group, a benzoxazinyl group, a β -carbolinyl group, a chromanyl group, a chromonyl group, a cinnolinyl group, a coumarinyl group, a decahydroquinolinyl group, a decahydroisoquinolinyl group, a dihydrobenzisothiazinyl group, a dihydrobenzisoxazinyl group, a dihydrofuranyl group, a dihydroisoindolyl group, a dihydropyranyl group, a dihydropyrazolyl group, a dihydropyrazinyl group, a dihydropyridinyl group, a dihydropyrimidyl group, a dihydropyrrolyl group, a dioxolanyl group, a1, 4-dithianyl group, a furanonyl group, an imidazolidinyl group, a2, 4-dioxo-imidazolidinyl group, an imidazolinyl group, an indolinyl group, a 2-oxo-indolinyl group, an isobenzotetrahydrofuranyl group, an isobenzotetrahydrothiophenyl group, isochromanyl, isocoumarin, isoindolinyl (isoindolinyl), 1-oxo-isoindolinyl, 1, 3-dioxo-isoindolinyl, isothiazolidinyl, isoxazolidinyl, 3-oxo-isoxazolidinyl, morpholinyl, 3, 5-dioxo-morpholinyl, octahydroindolyl, octahydroisoindolyl, 1-oxo-octahydroisoindolyl, 1, 3-dioxo-hexahydroisoindolyl, oxazolidinone, oxazolidinyl, oxiranyl, piperazinyl, 2, 6-dioxo-piperazinyl, piperidinyl, 2, 6-dioxo-piperidinyl, 4-piperidonyl, 2-oxopyrrolidinyl, 2, 5-dioxopyrrolidinyl, quinuclidinyl, tetrahydroisoquinolinyl, 3, 5-dioxo-thiomorpholinyl, thiazolidinyl, 2, 4-dioxo-thiazolidinyl, tetrahydroquinolinyl, phenothiazinyl, phenoxazinyl, xanthenyl and 1,3, 5-trithiohexanyl. In heterocyclic radicals of-CH₂Examples of-groups substituted by-C (═ O) -include, but are not limited to, 2-oxopyrrolidinyl, oxo-1, 3-thiazolidinyl, 2-piperidinonyl, 3, 5-dioxopiperidinyl and pyrimidinedione. Examples of the sulfur atom in the heterocyclic group being oxidized include, but are not limited to, sulfolane group, 1-dioxothiomorpholinyl group. The heterocyclyl group may be optionally substituted with one or more substituents as described herein.

In one embodiment, heterocyclyl is a 3-8 membered heterocyclyl, meaning that it contains 3-8 ring atomsWherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms. Unless otherwise specified, a heterocyclic group of 3 to 8 atoms may be carbon-based or nitrogen-based, and-CH₂-the group may optionally be replaced by-C (═ O) -. The sulfur atom of the ring may optionally be oxidized to the S-oxide. The nitrogen atom of the ring may optionally be oxidized to an N-oxygen compound. Examples of heterocyclic groups consisting of 3 to 8 atoms include, but are not limited to: azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, 1, 3-dioxolanyl, dithiocyclopentyl, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, dioxanyl, dithianyl, thioxanyl, homopiperazinyl, homopiperidinyl, oxepanyl, thiepanyl, oxacycloheptanyl, oxazepanyl, thiazepanyl, and the like

Radical, diaza

Radical, S-N-aza

And (4) a base. In heterocyclic radicals of-CH₂Examples of-groups substituted by-C (═ O) -include, but are not limited to, 2-oxopyrrolidinyl, oxo-1, 3-thiazolidinyl, 2-piperidinonyl, 3, 5-dioxopiperidinyl and pyrimidinedione. Examples of the sulfur atom in the heterocyclic group being oxidized include, but are not limited to, sulfolane group, 1-dioxothiomorpholinyl group. Said heterocyclyl group of 3 to 8 atoms may be optionally substituted by one or more substituents as described herein.

In one embodiment, heterocyclyl is a 3-6 atom heterocyclyl and refers to a saturated or partially unsaturated monocyclic ring containing 3-6 ring atoms in which at least one ring atom is selected from nitrogen, sulfur, and oxygen atoms. Unless otherwise statedIn addition, the heterocyclic group consisting of 3 to 6 atoms may be a carbon group or a nitrogen group, and-CH₂-the group may optionally be replaced by-C (═ O) -. The sulfur atom of the ring may optionally be oxidized to the S-oxide. The nitrogen atom of the ring may optionally be oxidized to an N-oxygen compound. Said heterocyclyl group of 3 to 6 atoms may be optionally substituted by one or more substituents as described herein.

In another embodiment, heterocyclyl is a 5-6 atom heterocyclyl and refers to a saturated or partially unsaturated monocyclic ring containing 5-6 ring atoms in which at least one ring atom is selected from the group consisting of nitrogen, sulfur, and oxygen atoms. Unless otherwise specified, a heterocyclic group of 5 to 6 atoms may be carbon-based or nitrogen-based, and-CH₂-the group may optionally be replaced by-C (═ O) -. The sulfur atom of the ring may optionally be oxidized to the S-oxide. The nitrogen atom of the ring may optionally be oxidized to an N-oxygen compound. Examples of heterocyclic groups consisting of 5 to 6 atoms include, but are not limited to: pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, 1, 3-dioxolanyl, dithiocyclopentyl, 2-oxopyrrolidinyl, oxo-1, 3-thiazolidinyl, sulfolane, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, dioxanyl, dithianyl, thiaxanyl, 2-piperidinonyl, 3, 5-dioxopiperidinyl and pyrimidinedione, 1-dioxothiomorpholinyl. Said heterocyclyl group of 5 to 6 atoms may be optionally substituted by one or more substituents as described herein.

The term "heterocyclylalkyl" includes heterocyclyl-substituted alkyl groups; the term "heterocyclylalkoxy" includes heterocyclyl-substituted alkoxy groups in which an oxygen atom is attached to the remainder of the molecule; the term "heterocyclylalkylamino" includes heterocyclyl-substituted alkylamino groups in which the nitrogen atom is attached to the remainder of the molecule. Where heterocyclyl, alkyl, alkoxy and alkylamino all have the meanings as described herein, such examples include, but are not limited to, azetidin-1-ylmethyl, azetidin-1-ylethyl, azetidin-1-ylpropyl, pyrrol-1-ylmethyl, pyrrol-1-ylethyl, pyrrol-1-ylpropyl, morpholin-4-ylethyl, piperazin-4-ylethyl, piperidin-4-ylethylamino and the like.

The terms "fused bicyclic ring", "fused bicyclic group", "fused ring group" denote a saturated or unsaturated fused ring system, referring to a non-aromatic bicyclic ring system, as shown in formula (a1), i.e., ring B shares a bond with ring B'. Such systems may contain independent or conjugated unsaturation, but the core structure does not contain aromatic or heteroaromatic rings (although aromatics may be substituents thereon). Each ring in the fused bicyclic ring is either a carbocyclic or a heteroalicyclic, examples of which include, but are not limited to, hexahydro-furo [3,2-b ] compounds]Furan, 2,3,3a,4,7,7 a-hexahydro-1H-indene, 7-azabicyclo [2.3.0 ]]Heptane, fused bicyclo [3.3.0]Octane, fused bicyclo [3.1.0]Hexane, these are contained within a fused bicyclic ring. And the fused bicyclic group may be substituted or unsubstituted, wherein the substituent may be, but is not limited to, D, F, Cl, Br, I, N₃、CN、NO₂、OH、SH、NH₂Oxo, alkyl, haloalkyl, cyano-substituted alkyl, hydroxyalkyl, alkoxy, haloalkoxy, haloalkoxyalkyl, or the like.

The term "fused heterobicyclic group" denotes a saturated or unsaturated fused ring system or bridged ring system, involving a non-aromatic bicyclic ring system or bridged ring system. Such systems may contain independent or conjugated unsaturation, but the core structure does not contain aromatic or heteroaromatic rings (although aromatics may be substituents thereon). And at least one ring system comprises one or more heteroatoms, wherein each ring system comprises a 3-7 membered ring, i.e. comprising 1-6 carbon atoms and 1-3 heteroatoms selected from N, O, P, S, whereby S or P is optionally substituted by one or more oxygen atoms to give, for example, SO,SO₂，PO，PO₂examples of such include, but are not limited to hexahydro-furo [3,2-b ]]Furan, 7-azabicyclo [2.3.0]Heptane, 2-azabicyclo [2.2.1]Heptane, octahydropyrrole [3,2-b ] and]pyrrole, octahydropyrrole [3,4-c ]]Pyrrole, octahydro-1H-pyrrole [3,2-b]Pyridine, and the like. And the fused heterobicyclic group may be substituted or unsubstituted, wherein the substituent may be, but is not limited to, D, F, Cl, Br, I, N₃、CN、NO₂、OH、SH、NH₂Oxo, alkyl, haloalkyl, cyano-substituted alkyl, hydroxyalkyl, alkoxy, haloalkoxy, haloalkoxyalkyl, or the like.

The terms "spirocyclic", "spiro", "spirobicyclic group", "spirobicyclic ring" indicate that one ring originates from a particular cyclic carbon on the other ring. For example, ring a and ring B share a carbon atom in two saturated ring systems, which are referred to as "spirocycles. Each ring within the spiro ring is either a carbocyclic or a heteroalicyclic. Examples include, but are not limited to, 2, 7-diazaspiro [4.4 ]]Nonan-2-yl, 7-oxo-2-azaspiro [4.5 ]]Decan-2-yl, 4-azaspiro [2.4 ]]Heptane-5-yl, 4-oxaspiro [2.4 ]]Heptane-5-yl, 5-azaspiro [2.4 ]]Heptane-5-yl, spiro [2.4 ]]Heptylalkyl, spiro [4.4 ]]Nonanyl, 7-hydroxy-5-azaspiro [2.4 ]]Heptane-5-yl, and the like. And said spirobicyclic group may be substituted or unsubstituted, wherein the substituents may be, but are not limited to, D, F, Cl, Br, I, N₃、CN、NO₂、OH、SH、NH₂Oxo, alkyl, haloalkyl, cyano-substituted alkyl, hydroxyalkyl, alkoxy, haloalkoxy, haloalkoxyalkyl, or the like.

The term "spirobicyclic group" means a spirobicyclic group having two points of attachment to the rest of the molecule, wherein the spirobicyclic group has the definition as described herein.

The term "spiroheterobicyclic group" means that one ring originates from a specific ring on another ringAs carbon. For example, as described above, ring a and ring B share a carbon atom in two saturated ring systems, and are referred to as "spirocycles. And at least one ring system comprises one or more heteroatoms, wherein each ring system comprises a 3-7 membered ring, i.e. comprising 1-6 carbon atoms and 1-3 heteroatoms selected from N, O, P, S, whereby S or P is optionally substituted by one or more oxygen atoms to give, for example, SO₂，PO，PO₂Examples of such include, but are not limited to, 4-azaspiro [2.4 ]]Heptane-5-yl, 4-oxaspiro [2.4 ]]Heptane-5-yl, 5-azaspiro [2.4 ]]Heptane-5-yl, 7-hydroxy-5-azaspiro [2.4 ]]Heptane-5-yl, 2, 6-diazaspiro [3.3]Heptane, 2, 6-diazaspiro [3.4 ]]Octane, 1, 6-diazaspiro [3.4 ]]Octane, 2, 7-diazaspiro [3.5 ]]Nonane, 1, 7-diazaspiro [3.5 ]]Nonane, 3, 9-diazaspiro [5.5 ]]Undecane, and the like. And the spiroheterobicyclic group may be substituted or unsubstituted, wherein the substituent may be, but is not limited to, D, F, Cl, Br, I, N₃、CN、NO₂、OH、 SH、NH₂Oxo, alkyl, haloalkyl, cyano-substituted alkyl, hydroxyalkyl, alkoxy, haloalkoxy, haloalkoxyalkyl, or the like.

The term "bridged ring group" as used herein denotes a saturated or unsaturated bridged ring system, relating to a non-aromatic bridged ring system, as shown in formula (a2), i.e. ring a1 shares an alkyl or a heteroalkyl chain with ring a2, wherein j is 1,2,3 or 4. Such systems may contain independent or conjugated unsaturation, but the core structure does not contain aromatic or heteroaromatic rings (although aromatics may be substituents thereon). Each ring in the bridged ring is either a carbocyclic or a heteroalicyclic, examples of which include, but are not limited to, bicyclo [2.2.1]Heptane, 2-azabicyclo [2.2.1]Heptane, 1,2,3,4,4a,5,8,8 a-octahydronaphthalene, which are contained within a fused bicyclic or bridged ring system. And the bridging group may be substituted or unsubstituted, wherein the substituents may be, but are not limited to, D, F, Cl, Br, I, N₃、 CN、NO₂、OH、SH、NH₂Oxo, alkyl, haloalkyl, cyano-substituted alkyl, hydroxyalkyl, alkoxy, haloAlkoxy, or haloalkoxyalkyl, and the like.

The term "bridged heterocyclyl" denotes a saturated or unsaturated bridged ring system, involving a non-aromatic bridged ring system. Such systems may contain independent or conjugated unsaturation, but the core structure does not contain aromatic or heteroaromatic rings (although aromatics may be substituents thereon). And at least one ring system comprises one or more heteroatoms, wherein each ring system comprises a 3-7 membered ring, i.e. comprising 1-6 carbon atoms and 1-3 heteroatoms selected from N, O, P, S, whereby S or P is optionally substituted by one or more oxygen atoms to give, for example, SO₂，PO，PO₂Examples of such include, but are not limited to, 2-azabicyclo [2.2.1 ]]Heptane, (1R,5S) -3, 6-diazabicyclo [3.1.1]Heptane, 2, 5-diazabicyclo [2.2.1]Heptane, (1R,5S) -8-azabicyclo [3.2.1]Octane, and the like. And the bridged heterocyclic group may be substituted or unsubstituted, wherein the substituents may be, but are not limited to, D, F, Cl, Br, I, N₃、CN、NO₂、OH、 SH、NH₂Oxo, alkyl, haloalkyl, cyano-substituted alkyl, hydroxyalkyl, alkoxy, haloalkoxy, haloalkoxyalkyl, or the like.

As described herein, there are two attachment points in the ring system that are attached to the rest of the molecule, as shown in formula (a3) or (a4), meaning that either the E or E' end is attached to the rest of the molecule, i.e., the attachment of the two ends can be interchanged.

E -C(＝O)-CH₂-O- E′ (a4)

The term "n-atomic" where n is an integer typically describes the number of ring-forming atoms in a molecule in which the number of ring-forming atoms is n. For example, piperidinyl is a heterocycloalkyl group of 6 atoms, while 1,2,3, 4-tetrahydronaphthalene is a cycloalkyl group of 10 atoms.

The term "heteroatom" refers to O, S, N, P and Si, including N, S and any oxidation state form of P; primary, secondary, tertiary amines and quaternary ammonium salt forms; or a form in which a hydrogen on a nitrogen atom in the heterocycle is substituted, for example, N (like N in 3, 4-dihydro-2H-pyrrolyl), NH (like NH in pyrrolidinyl) or NR (like NR in N-substituted pyrrolidinyl).

The term "cyano-substituted alkyl" or "cyanoalkyl" includes C substituted with one or more cyano groups_1-10A straight or branched alkyl group. In some of these embodiments, cyano-substituted alkyl is C substituted with one or more cyano groups_1-6"lower cyanoalkyl", other embodiments are where the cyano-substituted alkyl is C substituted with one or more cyano groups_1-4"lower cyanoalkyl", examples of which include, but are not limited to, CNCH₂-、CNCH₂CH₂-、CNCH₂CH₂CH₂-、CNCH₂CHCNCH₂-and the like.

As described herein, a ring system formed by a substituent on a ring having a bond to the center (as shown below) represents that the substituent may be substituted at any substitutable position on either ring. For example, formula B represents that any possible substituted position on the A or B ring may be substituted as shown in formulas c, d, e, f, g, h, i, j, k, l, m, n, o, p, q, etc.

As used herein, "pharmaceutically acceptable salts" refer to organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as are: berge et al, description of the scientific acceptable salts in detail in J. Pharmaceutical Sciences,1977,66:1-19. Pharmaceutically acceptable non-toxic acid salts include, but are not limited to, those formed by reaction with amino groupsInorganic acid salts such as hydrochloride, hydrobromide, phosphate, sulphate, perchlorate, and organic acid salts such as acetate, oxalate, maleate, tartrate, citrate, succinate, malonate, or by other methods described in the literature above, such as ion exchange. Other pharmaceutically acceptable salts include adipates, alginates, ascorbates, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, cyclopentylpropionates, digluconates, dodecylsulfates, ethanesulfonates, formates, fumarates, glucoheptonates, glycerophosphates, gluconates, hemisulfates, heptanoates, hexanoates, hydroiodides, 2-hydroxy-ethanesulfonates, lactobionates, lactates, laurates, malates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, palmitates, pamoates, pectinates, persulfates, 3-phenylpropionates, picrates, pivalates, propionates, stearates, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Salts obtained by reaction with a suitable base include alkali metals, alkaline earth metals, ammonium and N⁺(C₁-C₄Alkyl radical)₄A salt. The present invention also contemplates quaternary ammonium salts formed from compounds containing groups of N. Water-soluble or oil-soluble or dispersion products can be obtained by quaternization. Alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include suitable, non-toxic ammonium, quaternary ammonium salts and amine cations resistant to formation of counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C_1-8Sulfonates and aromatic sulfonates.

"solvate" of the present invention refers to an association of one or more solvent molecules with a compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to an association of solvent molecules that is water.

When the solvent is water, the term "hydrate" may be used. In some embodiments, a molecule of a compound of the present invention may be associated with a molecule of water, such as a monohydrate; in other embodiments, one molecule of the compound of the present invention may be associated with more than one molecule of water, such as a dihydrate, and in still other embodiments, one molecule of the compound of the present invention may be associated with less than one molecule of water, such as a hemihydrate. It should be noted that the hydrates of the present invention retain the biological effectiveness of the compound in its non-hydrated form.

The term "treating" or "treatment" as used herein refers, in some embodiments, to ameliorating a disease or disorder (i.e., slowing or arresting or reducing the development of the disease or at least one clinical symptom thereof). In other embodiments, "treating" or "treatment" refers to moderating or improving at least one physical parameter, including physical parameters that may not be perceived by the patient. In other embodiments, "treating" or "treatment" refers to modulating the disease or disorder, either physically (e.g., stabilizing a perceptible symptom) or physiologically (e.g., stabilizing a parameter of the body), or both. In other embodiments, "treating" or "treatment" refers to preventing or delaying the onset, occurrence, or worsening of a disease or disorder.

Disclosure of Invention

The aromatic heterocyclic compound provided by the invention can effectively inhibit ATX activity, and can be used for preparing medicaments for treating diseases with pathological characteristics of increased ATX expression, such as cancers, fibrotic diseases (such as pulmonary fibrosis or hepatic fibrosis), metabolic diseases, myelodysplastic syndrome, cardiovascular diseases, autoimmune diseases, inflammation, nervous system diseases or pain.

In one aspect, the present invention provides a novel heteroaromatic compound having a structure represented by formula (I), or (II):

wherein,

Cy¹is C_5-12Spiro bicyclic group, C_5-12Spiro-heterobicyclic radical, C_5-12Condensed bicyclic group, C_5-12Fused heterobicyclic radical, C_5-12Bridged ring radical, or C_5-12Bridged heterocyclyl, wherein Cy is¹Optionally substituted by 1,2,3 or 4R⁴Substitution;

Cy²is a heterocyclic group consisting of 3 to 8 atoms, wherein said Cy²Optionally substituted by 1,2,3 or 4R⁴Substitution;

Y¹is-C (═ O) -, or-S (═ O)_1-2-；

Y²Is- (CH)₂)_1-2-O-、-C(＝O)-CH₂-O-、-NHC(＝O)-CH₂-、-C(＝O)-(CH₂)_1-2-、-C(＝O)-CH₂-N(R^1b) -, or-NHC (═ O) NH-;

z is C_3-6Heterocyclyl radical C_1-4Alkyl, or

R^1aIs H, C_1-4Alkyl, or halo C_1-4An alkyl group;

R^1bis C_1-4alkyl-C (═ O) -, C_1-4alkyl-S (═ O)_1-2-、(C_1-4Alkyl radical)₂N-C (═ O) -, or C_2-7heterocyclyl-C (═ O) -C_1-4Alkyl-, said R^1bOptionally substituted by 1,2,3 or 4R⁷Substitution;

R²is H, -CN, -NO₂、-OH、-NH₂、F、Cl、Br、I、C_1-4Alkyl, halo C_1-4Alkyl, or C_1-4A hydroxyalkyl group;

R³is H, -CN, -NO₂、-OH、-NH₂、F、Cl、Br、I、C_1-4Alkyl, halo C_1-4Alkyl, cyano-substituted C_1-4Alkyl, or C_1-4A hydroxyalkyl group;

each R⁴Each independently H, D, oxo (C ═ O), -CN,-NO₂、-OH、-NH₂、F、Cl、Br、I、C_1-4Alkyl, halo C_1-4Alkyl, cyano-substituted C_1-4Alkyl radical, C_1-4Hydroxyalkyl radical, C_1-4Alkoxy, halo C_1-4Alkoxy, or halo C_1-4Alkoxy radical C_1-4An alkyl group;

each R⁵Each independently H, D, oxo (C ═ O), -CN, -NO₂、-OH、-NH₂、F、Cl、Br、I、C_1-4Alkyl, halo C_1-4Alkyl, cyano-substituted C_1-4Alkyl radical, C_1-4Hydroxyalkyl radical, C_1-4Alkoxy, halo C_1-4Alkoxy, or halo C_1-4Alkoxy radical C_1-4An alkyl group;

each R⁶Each independently H, D, oxo (C ═ O), -CN, -NO₂、-OH、-NH₂、F、Cl、Br、I、C_1-4Alkyl, halo C_1-4Alkyl, cyano-substituted C_1-4Alkyl radical, C_1-4Hydroxyalkyl radical, C_1-4Alkoxy, halo C_1-4Alkoxy, or halo C_1-4Alkoxy radical C_1-4An alkyl group;

each R⁷Each independently H, D, oxo (C ═ O), -CN, -NO₂、-OH、-NH₂、F、Cl、Br、I、C_1-4Alkyl, halo C_1-4Alkyl, cyano-substituted C_1-4Alkyl radical, C_1-4Hydroxyalkyl radical, C_1-4Alkoxy, halo C_1-4Alkoxy, or halo C_1-4Alkoxy radical C_1-4An alkyl group;

X⁶is N, or CH₂；

X⁷is-O-, -S-, -NH-, - (CH)₂)_m4-NH-(CH₂)_m5-、-(CH₂)_m4-O-(CH₂)_m5-、-(CH₂)_m4-S-(CH₂)_m5-, or- (CH)₂)_m6-；

Each m4 is independently 1,2,3 or 4;

each m5 is independently 0, 1,2,3, or 4;

each m6 is independently 1,2,3 or 4;

n2 is 0, 1,2,3 or 4; and

t is 0, 1,2,3 or 4;

or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, nitrogen oxide, metabolite, prodrug, or mixture thereof.

In some embodiments, the compounds of the present invention have a structure represented by formula (Ia), or (IIa):

In some embodiments, wherein Cy is¹Is that

Cy²Is that

Wherein,

X³、X⁴and X⁵Each independently being-O-, -S-, -NH-, - (CH)₂)_m1-NH-(CH₂)_m2-、-(CH₂)_m1-O-(CH₂)_m2-、-(CH₂)_m1-S-(CH₂)_m2-, or- (CH)₂)_m3-；

Each m1 is independently 1,2, or 3;

each m2 is independently 0, 1,2, or 3;

each m3 is independently 1,2, or 3; and

n1 is 0, 1,2,3 or 4.

In some embodiments, wherein Cy is²Is that

Wherein m3 is 1,2, or 3, and n1 is 0, 1,2,3, or 4.

In some embodiments, wherein Cy is¹Is that

Cy²Is that

Wherein, the Cy is¹And Cy²Independently optionally substituted by 1,2,3 or 4R⁴And (4) substitution.

In some embodiments, wherein Z is

In some embodiments, wherein R²Is H, -CN, -NO₂、-OH、-NH₂F, Cl, Br, I, methyl, ethyl, propyl, isopropyl, butyl, hydroxymethyl, hydroxyethyl, trifluoromethyl, or trifluoroethyl.

In some embodiments, wherein R^1bIs CH₃-C(＝O)-、CH₃CH₂-C(＝O)-、CH₃-S(＝O)_1-2-、CH₃CH₂-S(＝O)_1-2-、 (CH₃)₂N-C (═ O) -, or azetidinyl-C (═ O) -CH₂-, said R^1bOptionally substituted by 1,2,3 or 4R⁷Substitution; and

each R⁷Each independently is H, oxo (C ═ O), -CN, -NO₂、-OH、-NH₂、F、Cl、Br, I, methyl, ethyl, propyl, isopropyl, tert-butyl, methoxy, ethoxy, -OCH₂CF₃、-CF₃、-CH₂CF₃、-CH₂CH₂CN、-CH₂CH₂OH, or CHF₂-O-CH₂-。

In some embodiments, wherein each R is⁴、R⁵And R⁶Each independently is H, oxo (C ═ O), -CN, -NO₂、-OH、-NH₂F, Cl, Br, I, methyl, ethyl, propyl, isopropyl, tert-butyl, methoxy, ethoxy, -OCH₂CF₃、-CF₃、-CH₂CF₃、-CH₂CH₂CN、 CHF₂-O-CH₂-、CF₃-O-CH₂-, or-CH₂CH₂OH。

In some embodiments, the compounds of the present invention are compounds having one of the following structures:

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, nitrogen oxide, metabolite, prodrug thereof, and a pharmaceutically acceptable adjuvant, diluent, or carrier.

In some embodiments, the pharmaceutical compositions of the present invention further comprise an additional therapeutic agent.

In some embodiments, the composition of the invention wherein the additional therapeutic agent is a therapeutic agent for a disease associated with a fibrotic disease, a proliferative disease, an inflammatory disease, an autoimmune disease, a respiratory disease, a cardiovascular disease, a neurodegenerative disease, a dermatological disorder, and/or abnormal angiogenesis.

In some embodiments, the pharmaceutical composition of the present invention, wherein the additional therapeutic agent includes, but is not limited to, immunomodulators, analgesics, non-steroidal anti-inflammatory drugs, steroids, synthetic DMARDS, drugs for treating proliferative diseases, glucocorticoids, cytostatics, alkylating agents, antimetabolites, cytotoxic antibiotics, antibodies, and the like.

In another aspect, the present invention provides a use of a compound of the present invention or a pharmaceutical composition of the present invention for the manufacture of a medicament for preventing or treating a disease characterized by a pathology in which ATX expression is increased in a mammal.

In some embodiments, wherein the disease having a pathological feature of increased ATX expression comprises: cancer, fibrotic disease, metabolic disease, myelodysplastic syndrome, cardiovascular disease, autoimmune disease, inflammation, neurological disease, or pain.

In some embodiments, wherein the disease with a pathological feature of increased ATX expression is pulmonary fibrosis or liver fibrosis.

In some embodiments, a compound of the invention or a pharmaceutical composition thereof may be administered in combination with an additional therapeutic agent.

In some embodiments, the use of the invention comprises administering to a mammal an amount of a compound or pharmaceutical composition of the invention sufficient to effect said treatment or prevention.

Pharmaceutical composition, preparation and use

When used as a medicament, the compounds of the present invention are typically administered in the form of a pharmaceutical composition. The compositions may be prepared in a manner well known in the pharmaceutical art and comprise at least one compound according to the invention according to formula I, Ia, II, or IIa. Typically, the compounds of the present invention are administered in a pharmaceutically effective amount. The amount of the compound of the invention actually administered will generally be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound of the invention administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, and the like.

The pharmaceutical compositions of the present invention may be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intra-articular, intravenous, intramuscular, and intranasal. Depending on the intended route of delivery, the compounds of the invention are preferably formulated as injectable or oral compositions or as ointments, as lotions or as patches (all for transdermal administration).

As liquid compositions for oral administration, pharmaceutically acceptable solutions, suspensions, emulsions, syrups and elixirs containing inert diluents such as water or liquid paraffin may be used. Such compositions may also include substances other than diluents, and in some embodiments, humectants, sweeteners, or flavoring agents.

Compositions for parenteral administration may be emulsions or sterile solutions. In certain embodiments, propylene glycol, polyethylene glycol, vegetable oils, particularly olive oil, or injectable organic esters may be used as a solvent or carrier, and in some embodiments, ethyl oleate may be used as a solvent or carrier. These compositions may also comprise adjuvants, in particular wetting agents, isotonicity agents, emulsifiers, dispersants and stabilizers. Sterilization can be performed in several ways, in certain embodiments, using bacteriological filters, by radiation or by heating. They may also be prepared in the form of sterile solid compositions which may be dissolved in sterile water or any other injectable sterile medium at the time of use.

The composition may also be an aerosol. For use in the form of a liquid aerosol, the composition may be a stable sterile solution or a solid composition dissolved in pyrogen-free sterile water, saline, or any other pharmaceutically acceptable carrier at the time of use. For use in the form of a dry aerosol intended for direct inhalation, the active ingredients are finely divided and combined with a water-soluble solid diluent or carrier, in certain embodiments, dextran, mannitol or lactose.

Typical pharmaceutical compositions and dosage forms contain one or more excipients. Suitable excipients are well known to those skilled in the art of pharmacy and in certain embodiments include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. Whether a particular excipient is suitable for incorporation into a pharmaceutical composition or dosage form depends on a variety of factors well known in the art, including, but not limited to, the manner in which the dosage form is administered to a subject and the particular active ingredient in the dosage form. The compositions or single unit dosage forms may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents, if desired.

In another aspect, the present invention provides a compound of the invention or a pharmaceutical composition comprising a compound of the invention for use in medicine. In a specific embodiment, the present invention provides a compound of the present invention or a pharmaceutical composition comprising a compound of the present invention for use in the prevention and/or treatment of fibrotic diseases, proliferative diseases, inflammatory diseases, autoimmune diseases, respiratory diseases, cardiovascular diseases, neurodegenerative diseases, dermatological disorders, and/or abnormal angiogenesis-related diseases.

In some embodiments, the present invention provides a compound of the present invention or a pharmaceutical composition comprising a compound of the present invention for use in the preparation of a medicament for the prevention and/or treatment of a fibrotic disease, a proliferative disease, an inflammatory disease, an autoimmune disease, a respiratory disease, a cardiovascular disease, a neurodegenerative disease, a dermatological disorder, and/or an abnormal angiogenesis-related disease.

In some embodiments, the invention provides pharmaceutical compositions comprising a compound of the invention and an additional therapeutic agent. In particular embodiments, the additional therapeutic agent is a therapeutic agent for a disease associated with a fibrotic disease, a proliferative disease, an inflammatory disease, an autoimmune disease, a respiratory disease, a cardiovascular disease, a neurodegenerative disease, a dermatological disorder, and/or abnormal angiogenesis.

In some embodiments, the present invention provides a compound of the present invention or a pharmaceutical composition comprising a compound of the present invention for use in the prevention and/or treatment of a fibrotic disease. In particular embodiments, the fibrotic disease is selected from Idiopathic Pulmonary Fibrosis (IPF), cystic fibrosis, other diffuse parenchymal lung diseases of different etiology (including iatrogenic drug-induced fibrosis, occupational and/or environmentally induced fibrosis), granulomatous diseases (sarcoidosis, hypersensitivity pneumonitis), collagen vascular disease, pulmonary alveolar protein deposition, langerhans cell granuloma, lymphangioleiomyomatosis, genetic diseases (helminthependra syndrome, tuberous sclerosis, neurofibromas, metabolic accumulation disorders, familial interstitial lung disease), radiation-induced fibrosis, Chronic Obstructive Pulmonary Disease (COPD), scleroderma, bleomycin-induced pulmonary fibrosis, chronic asthma, silicosis, asbestos-induced pulmonary fibrosis, Acute Respiratory Distress Syndrome (ARDS), renal fibrosis, pulmonary fibrosis, and inflammatory bowel disease, Tubulointerstitial fibrosis, glomerulonephritis, regional segmental glomerulosclerosis, IgA nephropathy, hypertension, Alport disease (Alport), intestinal fibrosis, hepatic fibrosis, cirrhosis, alcohol-induced hepatic fibrosis, toxin/drug-induced hepatic fibrosis, hemochromatosis, non-alcohol steatohepatitis (NASH), bile duct injury, primary biliary cirrhosis, infection-induced hepatic fibrosis, virus-induced hepatic fibrosis and autoimmune hepatitis, corneal scarring, hypertrophic scarring, dipterland disease, keloids, skin fibrosis, cutaneous scleroderma, systemic sclerosis, spinal cord injury/fibrosis, bone marrow fibrosis, vascular restenosis, atherosclerosis, arteriosclerosis, wegener's granuloma, peloniosis, or chronic lymphocytic. More particularly, the fibrotic disease is Idiopathic Pulmonary Fibrosis (IPF).

In other embodiments, the present invention provides a compound of the present invention or a pharmaceutical composition comprising a compound of the present invention for use in the preparation of a medicament for the prevention and/or treatment of a fibrotic disease. In particular embodiments, the fibrotic disease is selected from Idiopathic Pulmonary Fibrosis (IPF), cystic fibrosis, other diffuse parenchymal lung diseases of different etiology (including iatrogenic drug-induced fibrosis, occupational and/or environmentally induced fibrosis), granulomatous diseases (sarcoidosis, hypersensitivity pneumonitis), collagen vascular disease, pulmonary alveolar protein deposition, langerhans cell granuloma, lymphangioleiomyomatosis, genetic diseases (helminth-primordia syndrome, sarcoidosis, neurofibromas, metabolic accumulation disorders, familial interstitial lung disease), radiation-induced fibrosis, Chronic Obstructive Pulmonary Disease (COPD), scleroderma, bleomycin-induced pulmonary fibrosis, chronic asthma, silicosis, asbestos-induced pulmonary fibrosis, Acute Respiratory Distress Syndrome (ARDS), Renal fibrosis, tubulointerstitial fibrosis, glomerulonephritis, regional segmental glomerulosclerosis, IgA nephropathy, hypertension, Alport disease (Alport), intestinal fibrosis, hepatic fibrosis, cirrhosis, alcohol-induced hepatic fibrosis, toxin/drug-induced hepatic fibrosis, hemochromatosis, nonalcoholic steatohepatitis (NASH), bile duct injury, primary biliary cirrhosis, infection-induced hepatic fibrosis, virus-induced hepatic fibrosis and autoimmune hepatitis, corneal scarring, hypertrophic scarring, diptertagian disease, epilepsy, dermal fibrosis, dermal scleroderma, systemic sclerosis, spinal cord injury/fibrosis, bone marrow fibrosis, vascular restenosis, atherosclerosis, arteriosclerosis, wegener granuloma, perlitic disease, or chronic lymphocytic fibrosis. More particularly, the fibrotic disease is Idiopathic Pulmonary Fibrosis (IPF).

In a further method of therapeutic aspects, the invention provides a method of preventing and/or treating a mammal having a fibrotic disease, the method comprising administering an effective amount of one or more of the compounds or pharmaceutical compositions described herein for treating or preventing the condition. In particular embodiments, the fibrotic disease is selected from Idiopathic Pulmonary Fibrosis (IPF), cystic fibrosis, other diffuse parenchymal lung diseases of different etiology (including iatrogenic drug-induced fibrosis, occupational and/or environmentally induced fibrosis), granulomatous diseases (sarcoidosis, hypersensitivity pneumonitis), collagen vascular disease, pulmonary alveolar protein deposition, langerhans cell granuloma, lymphangioleiomyomatosis, genetic diseases (helminth-primordia syndrome, sarcoidosis, neurofibromas, metabolic accumulation disorders, familial interstitial lung disease), radiation-induced fibrosis, Chronic Obstructive Pulmonary Disease (COPD), scleroderma, bleomycin-induced pulmonary fibrosis, chronic asthma, silicosis, asbestos-induced pulmonary fibrosis, Acute Respiratory Distress Syndrome (ARDS), Renal fibrosis, tubulointerstitial fibrosis, glomerulonephritis, regional glomerulosclerosis, IgA nephropathy, hypertension, Alport disease (Alport), intestinal fibrosis, hepatic fibrosis, cirrhosis, alcohol-induced hepatic fibrosis, toxin/drug-induced hepatic fibrosis, hemochromatosis, non-alcoholic steatohepatitis (NASH), bile duct injury, primary biliary cirrhosis, infection-induced hepatic fibrosis, virus-induced hepatic fibrosis and autoimmune hepatitis, corneal scarring, hypertrophic scarring, diptertron disease, keloids, skin fibrosis, cutaneous scleroderma, systemic sclerosis, spinal cord injury/fibrosis, bone marrow fibrosis, vascular restenosis, atherosclerosis, arteriosclerosis, wegener's granuloma, peroneal disease or chronic lymphocytic. More particularly, the fibrotic disease is Idiopathic Pulmonary Fibrosis (IPF).

A particular embodiment of the method of the invention comprises administering to an individual suffering from a fibrotic disease an effective amount of a compound of the invention of formula I, Ia, II, or IIa for a period of time sufficient to reduce the level of fibrosis in the individual and preferably to terminate the process that causes said fibrosis. Particular embodiments of the methods comprise administering to an individual patient having a developing idiopathic pulmonary fibrosis an effective amount of a compound of the present invention of formula I, Ia, II, or IIa for a period of time sufficient to reduce or prevent idiopathic pulmonary fibrosis in the patient, and preferably to terminate the process that causes the idiopathic pulmonary fibrosis.

As will be apparent to those skilled in the art, co-administration includes any manner of delivering two or more therapeutic agents to a patient as part of the same treatment regimen. While two or more active agents may be administered simultaneously in a single formulation (i.e., as a single pharmaceutical composition), this is not required. The active agents may also be administered in different formulations, at different times.

Detailed Description

To illustrate the invention, the following examples are set forth. It is to be understood that the invention is not limited to these embodiments, but is provided as a means of practicing the invention.

In general, the compounds of the invention can be prepared by the methods described herein, unless otherwise indicated, wherein the substituents are as defined in formula I, Ia, II, or IIa. The following reaction schemes and examples serve to further illustrate the context of the invention.

Those skilled in the art will recognize that: the chemical reactions described herein may be used to suitably prepare a number of other compounds of the invention, and other methods for preparing the compounds of the invention are considered to be within the scope of the invention. For example, the synthesis of those non-exemplified compounds according to the present invention can be successfully accomplished by those skilled in the art by modification, such as appropriate protection of interfering groups, by the use of other known reagents in addition to those described herein, or by some routine modification of reaction conditions. In addition, the reactions disclosed herein or known reaction conditions are also recognized as being applicable to the preparation of other compounds of the present invention.

The examples described below, unless otherwise indicated, are all temperatures set forth in degrees Celsius. Reagents were purchased from commercial suppliers such as Aldrich Chemical Company, Arco Chemical Company and Alfa Chemical Company and were used without further purification unless otherwise indicated. General reagents were purchased from Shantou Wen Long chemical reagent factory, Guangdong Guanghua chemical reagent factory, Guangzhou chemical reagent factory, Tianjin Haojian Yunyu chemical Co., Ltd, Tianjin Shucheng chemical reagent factory, Wuhan Xin Huayuan scientific and technological development Co., Ltd, Qingdao Tenglong chemical reagent Co., Ltd, and Qingdao Kaolingyi factory.

The anhydrous tetrahydrofuran, dioxane, toluene and ether are obtained through reflux drying of metal sodium. The anhydrous dichloromethane and chloroform are obtained by calcium hydride reflux drying. Ethyl acetate, petroleum ether, N-hexane, N, N-dimethylacetamide and N, N-dimethylformamide were used as they were previously dried over anhydrous sodium sulfate.

The following reactions are generally carried out under positive pressure of nitrogen or argon or by sleeving a dry tube over an anhydrous solvent (unless otherwise indicated), the reaction vial being stoppered with a suitable rubber stopper and the substrate being injected by syringe. The glassware was dried.

The column chromatography is performed using a silica gel column. Silica gel (300 and 400 meshes) was purchased from Qingdao oceanic chemical plants.

¹H NMR spectra were recorded using a Bruker 400MHz or 600MHz NMR spectrometer.¹H NMR Spectrum in CDC1₃、DMSO-d₆、 CD₃OD or acetone-d₆TMS (0ppm) or chloroform (7.26ppm) was used as a reference standard for the solvent (in ppm). When multiple peaks occur, the following abbreviations will be used: s (singleton), d (doublet), t (triplet), m (multiplet), br (broad), dd (doublet of doublets), dt (doublet of triplets). Coupling constants are expressed in hertz (Hz).

The conditions for determining low resolution Mass Spectrometry (MS) data were: agilent 6120 four-stage rod HPLC-M (column model: Zorbax SB-C18, 2.1X 30mm,3.5 micron, 6min, flow rate 0.6 mL/min. mobile phase: 5% -95% ((CH with 0.1% formic acid)₃CN) in (H containing 0.1% formic acid)₂O) by electrospray ionization (ESI) at 210nm/254nm, with UV detection.

Pure compounds were detected by UV at 210nm/254nm using Agilent 1260pre-HPLC or Calesep pump 250pre-HPLC (column model: NOVASEP 50/80 mm DAC).

A typical synthetic procedure for the preparation of the disclosed compounds is shown in scheme 1 below.

Synthesis scheme 1:

wherein E is¹And E²Each independently selected from Cl, Br or I; e³Selected from Cl, Br, I, OMs, OTs or OTf; pr (Pr) of¹Selected from Boc, Cbz or PMB; pr (Pr) of²Selected from tert-butyl or 2,4, 4-trimethylpent-2-yl; cy has the Cy of the present invention¹And Cy²The definition; y has the formula¹And Y²The definition; r^1a、R²、R³、R⁶Z and t all have the definitions given in the present invention.

Reacting intermediate 1-1 with Pr²NC and aldehyde R²CHO generates an intermediate 1-2 through three-component reaction under the catalysis of Lewis acid; reacting the intermediate 1-2 in formic acid under the condition of heating to obtain an intermediate 1-3; carrying out nucleophilic substitution reaction on the obtained intermediate 1-3 and the intermediate 2-2 under the conditions of alkalinity and heating condition to obtain an intermediate 1-4; the intermediate 1-4 is reacted with a strong base, then subjected to nucleophilic substitution with the intermediate 1-5, and then subjected to nucleophilic substitution with the substituted alkyl R^1aE⁴Reacting to obtain an intermediate 1-6; the intermediate 1-6 can be hydrogenated under acid condition or palladium catalysis or reacted with iodotrimethylsilane to remove protecting group Pr¹To give intermediates 1 to 7; and carrying out nucleophilic substitution reaction on the intermediates 1-7 and the intermediates 2-7 under alkaline and heating conditions to obtain the compound shown in the formula I or II.

Examples

Example 1

Preparation of 2- { [ 6-Ethyl-2- (3- ((2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) -1, 4-diazepan-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

Step 1) 2-bromo-6-ethyl-N- (2,4, 5-trimethylpentan-2-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-amine

To a mixture of 2-amino-5-bromo-1, 3, 4-thiadiazole (2.0g, 11.2mmol) in n-butanol (20mL) were added 1,1,3, 3-tetramethylbutylisonitrile (1.56g, 11.2mmol), n-propionaldehyde (1.6g, 28mmol) and anhydrous magnesium chloride (0.5g, 5.6mmol), and the resulting mixture was reacted at 120 ℃ for 4 hours, and the solvent was removed by concentration under reduced pressure. The resulting residue was purified by column chromatography (ethyl acetate: petroleum ether ═ 1:5) to give the title compound as a brown oil (3.5g, 88%). MS (m/z): 359.1[ M +1], 361.1[ M +1 ].

Step 2): n- (2-bromo-6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl) carboxamide

A solution of 2-bromo-6-ethyl-N- (2,4, 5-trimethylpentan-2-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-amine (3.5g, 9.75mmol) in formic acid (20mL) is reacted at 80 ℃ for 3 hours, concentrated under reduced pressure, and then petroleum ether is added to the residue for slurrying, filtration and drying to give 2.3g of product, yield: 74 percent. MS (m/z): 275.0[ M +1], 277.0[ M +1 ].

Step 3)2- { [ 2-bromo-6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To a mixture of N- (2-bromo-6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl) carboxamide (525mg, 1.91mmol) in THF (8mL) with cooling in an ice bath was added 60% sodium hydride (229mg, 5.73 mmol). After the resulting mixture was stirred for 15 minutes, a solution of 2-chloro-4- (4-fluorophenyl) thiazole-5-carbonitrile (454mg, 1.91mmol) in tetrahydrofuran (3mL) was added dropwise. The reaction was allowed to warm to room temperature for 30 minutes, the completion of the reaction was monitored on a thin-layer plate, and methyl iodide (542mg, 3.82mmol) was added to the reaction mixture, followed by reaction at room temperature for 2 hours. The reaction was detected to be complete by LC-MS, water was added dropwise to the mixture to quench the reaction, water (20mL) was then added, followed by extraction with ethyl acetate (15mL × 3), the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the resulting residue was purified by column chromatography (petroleum ether: ethyl acetate ═ 2:1) to give the title compound as a white solid (603mg, 68%).

Step 4)4- {5- [ (5-cyano-4- (4-fluorophenyl) thiazol-2-yl) amino ] -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl } -1, 4-diazepan-1-carboxylic acid tert-butyl ester

To a solution of tert-butyl 1, 4-diazepan-1-carboxylate (217mg, 1.08mmol) in DMF (6mL) was added potassium carbonate (450mg, 3.26mmol) and 2- { [ 2-bromo-6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (500 mg, 1.08 mmol). The reaction mixture was heated to 70 ℃ for 3 hours and checked by LC-MS for completion. The reaction solution was poured into water (20mL), extracted with ethyl acetate (20mL × 3), the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the resulting residue was purified by column chromatography (petroleum ether: ethyl acetate ═ 1:1) to give 561mg of a pale yellow solid, yield: 89 percent.

Step 5)2- { [2- (1, 4-diazepan-1-yl) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To 4- {5- [ (5-cyano-4- (4-fluorophenyl) thiazol-2-yl) amino group at 0-5 deg.C]-6-ethylimidazo [2,1-b][1,3,4]Thiadiazol-2-yl } -1, 4-diazepan-1-carboxylic acid tert-butyl ester (561mg, 0.96mmol) in DCM (10mL) was added dropwise with CF dropwise₃COOH (1mL), after 10 minutes, warmed to room temperature for 1 hour, and checked by LC-MS for completion. The mixture was concentrated under reduced pressure, the resulting residue was dissolved in MeOH (20mL), NaHCO was added₃(2g) After stirring and neutralization for 30min, concentrated under reduced pressure, the residue was dissolved by addition of DCM (20mL), filtered and concentrated under reduced pressure to give the title compound as a pale yellow solid (456mg, 98%).

Step 6)2- { [ 6-Ethyl-2- (3- ((2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) -1, 4-diazepan-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To 2- { [2- (1, 4-diazepan-1-yl) -6-ethylimidazo [2,1-b][1,3,4]Thiadiazol-5-yl]To a solution of (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (80mg, 0.17mmol) in acetonitrile (5mL) was added K in sequence₂CO₃(70mg, 0.51mmol) and KI (14mg, 0.8mmol), and after stirring the mixture for 5 minutes, 2-chloro-1- (3-hydroxyazetidin-1-yl) ethanone (25mg, 0.17mmol) was added. The mixture was heated to 75 ℃ for 2 hours, filtered, concentrated under reduced pressure, and the resulting residue was purified by column chromatography (dichloromethane: methanol 15:1) to give the title compound as a white solid (45mg,46％)。¹H NMR(400MHz,CDCl₃)δ:8.14(m,2H),7.16(m,2H),4.64(m,1H), 4.35(m,1H),4.25(m,1H),4.01(m,1H),3.87(m,1H),3.66(m,2H),3.59-3.56(m,5H),3.22(m,2H),2.93(m,2H), 2.82(m,2H),2.58(q,J＝7.6Hz,2H),2.02(m,2H),1.27(m,3H)。MS(m/z)：596.4[M+1]。

example 2

Preparation of (R) -N- {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl ] pyrrolidin-3-yl } -N- (2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) acetamide

Step 1 tert-butyl (R) - {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl ] pyrrolidin-3-yl } carboxylate

To a solution of (R) -3-tert-butoxyamidopyrrolidine (0.98g, 5.23mmol) in DMF (15mL) were added potassium carbonate (1.97g, 14.3mmol) and 2- { [ 2-bromo-6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (2.2g, 4.76 mmol). The mixture was heated to 65 ℃ for 3 hours and checked by LC-MS for completion. The reaction solution was poured into water (30mL), extracted with ethyl acetate (30mL × 3), the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the resulting residue was purified by column chromatography (petroleum ether: ethyl acetate ═ 1.5:1) to give the title compound as a white solid (2.61g, 97%).

Step 2) (R) -2- { [2- (3-Aminopyrrolidin-1-yl) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To (R) - {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2, 1-b) at 0-5 deg.C][1,3,4]Thiadiazol-2-yl]A stirred solution of t-butyl pyrrolidin-3-yl } carboxylate (2.61g, 4.59mmol) in DCM (10mL) was added CF dropwise₃COOH (1mL), after 10 minutes the reaction was warmed to room temperature for 1 hour, and the reaction was completed by LC-MS detection. The mixture was concentrated under reduced pressure, the resulting residue was dissolved in MeOH (20mL), NaHCO was added₃(3g) After stirring and neutralization for 30min, concentrated under reduced pressure, the residue was dissolved by addition of DCM (20mL), filtered and concentrated under reduced pressure to give the title compound as a white solid (1.89g, 88%).

Step 3) (R) -2- { [ 6-Ethyl-2- (3- ((2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) amino) tetrahydropyrrolidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To (R) -2- { [2- (3-aminopyrrolidin-1-yl) -6-ethylimidazo [2,1-b][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (1.5g, 3.2mmol) in acetonitrile (28mL) was added K in sequence₂CO₃(1.32g, 9.6mmol) and KI (0.27g, 1.6mmol) and after stirring the resulting mixture for 5 minutes, 2-chloro-1- (3-hydroxyazetidin-1-yl) ethanone (0.58g, 3.85mmol) was added. The mixture was heated to 65 ℃ for 2 hours, filtered, concentrated under reduced pressure, and the resulting residue was purified by column chromatography (dichloromethane: methanol 25:1) to give the title compound as a white solid (1.27g, 67%).¹H NMR(400MHz,CDCl₃)δ:8.15(m,2H),7.18(m,2H),4.70(m, 1H),4.30(m,2H),3.99(m,1H),3.91(m,1H),3.64-3.46(m,7H),3.33-3.16(m,3H),2.60(q,J＝7.6Hz,2H),2.22(m, 1H),1.99(m,2H),1.28(m,3H)。MS(m/z)：582.2[M+1]。

Step 4) (R) -N- {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl ] pyrrolidin-3-yl } -N- (2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) acetamide

To (R) -2- { [ 6-ethyl-2- (3- ((2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) amino) tetrahydropyrrolidin-1-yl) imidazo [2,1-b ] at 0 deg.C][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (70mg, 0.12mmol) in dichloromethane (5mL) was added dropwise AcCl (28mg, 0.36mmol) and the mixture was stirred at this temperature for 5 minutes, after which Et was slowly added dropwise₃N (37mg, 0.36 mmol). The mixture was allowed to warm to room temperature overnight, concentrated under reduced pressure, and the residue was taken up in THF (2mL) and water (2mL) and K was added₂CO₃(83mg, 0.6mmol), heated to 60 ℃ for overnight reaction, filtered, concentrated under reduced pressure and the resulting residue purified on thick prep plates (dichloromethane: methanol 15:1,) to give the title compound as a white solid (35mg, 47%).¹H NMR(400MHz,CDCl₃)δ:8.15(m, 2H),7.16(m,2H),4.69(m,1H),4.47(m,1H),4.28(m,1H),4.14(m,1H),3.98-3.40(m,12H),2.60(m,2H),2.39(m, 1H),2.39-2.22(m,4H),1.28(m,3H)。MS(m/z)：624.2[M+1]。

Example 3

Preparation of (R) -2- { [ -2- (3- (bis (2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) amino) pyrrolidin-1-yl) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To (R) -2- { [ 6-ethyl-2- (3- ((2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) amino) tetrahydropyrrolidin-1-yl) imidazo [2,1-b][1,3,4]Thiadiazol-5-yl]To a solution of (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (1.08g, 1.86mmol) in MeCN (20mL) was added K in sequence₂CO₃(0.77g, 5.57mmol), KI (0.15g, 0.93mmol) and 2-chloro-1- (3-hydroxyazetidin-1-yl) ethanone (0.33g, 2.23mmol) and the mixture was allowed to warm to 90 ℃ for reaction overnight. The reaction solution was cooled to room temperature, filtered, concentrated under reduced pressure, and the resulting residue was purified by column chromatography (dichloromethane: methanol ═ 20:1) to give the title compound as a white solid (320mg, 25%).¹H NMR(400MHz, CDCl₃)δ:8.15(m,2H),7.16(m,2H),4.61(m,1H),4.53(m,1H),4.43-4.35(m,2H),4.25-4.20(m,3H),4.13(m,1H), 3.89-3.78(m,2H),3.68-3.58(m,5H),3.49-3.28(m,4H),3.17(m,1H),2.60(q,J＝7.6Hz,2H),2.25(m,1H),2.02(m, 4H),1.28(m,3H)。MS(m/z)：695.1[M+1]。

Example 4

Preparation of (R) -N- {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl ] pyrrolidin-3-yl } -N- (2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) methanesulfonamide

To (R) -2- { [ 6-ethyl-2- (3- ((2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) amino) tetrahydropyrrolidin-1-yl) imidazo [2,1-b at 0 deg.C][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (70mg, 0.12mmol) in DCM (5mL) was added TMSCl (39mg, 0.36mmol) dropwise, the mixture was kept at this temperature for stirring for 5 minutes, and after stirring, Et (Et) was slowly added dropwise₃N (74mg, 0.72mmol), and then reacted at 0 ℃ for 1 hour. The reaction mixture was cooled to-20 ℃ and MsCl (11mg, 0.10mmol) was added slowly and the reaction was maintained at-20 ℃ for 2 h. The reaction was checked for completion by LC-MS, quenched by addition of methanol, filtered, concentrated under reduced pressure, and the resulting residue was purified on a thick prep plate (dichloromethane: methanol 15:1) to give the title compound as a white solid (35mg, 44%).¹H NMR(400MHz, CDCl₃)δ:8.14(m,2H),7.16(m,2H),4.68(m,1H),4.59(m,1H),4.33(m,1H),4.25(m,1H),4.07-3.86(m,3H), 3.79-3.58(m,5H),3.45(m,2H),3.12(s,3H),2.61(m,2H),2.39(m,1H),2.20(m,1H),2.00(m,1H),1.26(m,3H)。 MS(m/z)：660.4[M+1]。

Example 5

Preparation of (R) -1- {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl ] pyrrolidin-3-yl } -3- (3-hydroxycyclobutane) urea

To a mixture of triphosgene (19mg, 0.064mmol) in DCM (2mL) was added dropwise (R) -2- { [2- (3-aminopyrrolidin-1-yl) -6-ethylimidazo [2,1-b ] under argon protection at 0 deg.C][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (60mg, 0.128 mmol) and triethylamine (53. mu.L, 0.38mmol) in DCM (1 mL). After the addition was complete, the mixture was stirred at 0 ℃ for 30 minutes, and 3-aminocyclobutanol hydrochloride (47mg, 0.38mmol) and tris (hydroxymethyl) aminomercyclobutanol hydrochloride (NMDA) were addedEthylamine (73. mu.L, 0.53mmol) in DMF (1 mL). The mixture was reacted at room temperature overnight, the reaction was poured into water (10mL), extracted with DCM (15mL x 3), the organic layers were combined, washed with saturated brine (20mL x 3), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the resulting residue was purified on thick prep plates (dichloromethane: methanol 15:1) to give the title compound as a white solid (42mg, 57%).¹H NMR(400MHz,CDCl₃)δ:8.13-8.05(m,2H),7.16-7.05(m, 2H),5.57(m,0.5H),5.24(m,0.5H),4.88(m,1H),4.54(m,1.5H),4.34(m,0.5H),4.08(m,1H),3.86-3.52(m,6H), 3.39-3.28(m,2H),2.81(m,1H),2.58(m,2H),2.37-2.17(m,3H),1.99(m,1H),1.81(m,1H),1.28(m,3H)。MS (m/z)：582.2[M+1]。

Example 6

Preparation of (S) -1- {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl ] pyrrolidin-3-yl } -3- (3-hydroxycyclobutane) urea

Referring to example 5, (S) -1- {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2,1-b ] was prepared using (S) -3-tert-butoxyamidopyrrolidine instead of (R) -3-tert-butoxyamidopyrrolidine][1,3,4]Thiadiazol-2-yl]Pyrrolidin-3-yl } -3- (3-hydroxycyclobutane) urea.¹H NMR(400MHz,CDCl₃)δ:8.13-8.06(m,2H),7.16-7.06(m,2H),5.57(m,0.5H), 5.26(m,0.5H),4.96(m,1H),4.54(m,1H),4.08(m,1H),3.83-3.52(m,7H),3.41-3.28(m,2H),2.81(m,1H),2.57(m, 2H),2.35-2.22(m,3H),2.00(m,1H),1.81(m,1H),1.28(m,3H)。MS(m/z)：582.2[M+1]。

Example 7

Preparation of (R) -N- {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl ] pyrrolidin-3-yl } -1- (2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) -3, 3-dimethylurea

Step 1) (R) -2- { [2- (3- ((2- (3- ((tert-butyldimethylsilyl) oxy) azetidin-1-yl) -2-oxoethyl) amino) pyrrolidin-1-yl) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To an acetonitrile (10mL) mixture of (R) -2- { [2- (3-aminopyrrolidin-1-yl) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (280mg, 0.60mmol), 1- (3- ((tert-butyldimethylsilyl) oxy) azetidin-1-yl) -2-chloroacetophenone (210mg, 0.78mmol) were added potassium carbonate (165mg, 1.2mmol) and potassium iodide (50mg, 0.30mmol), the resulting mixture was heated under reflux for 4 hours, cooled, filtered, concentrated under reduced pressure, and the resulting residue was purified by column chromatography (dichloromethane: methanol ═ 30:1) to give the title compound as a yellow foamy solid (300mg, 72%). LC-MS (M/z):696.3[ M +1 ].

Step 2) step 1) (R) -2- { [2- (3- ((2- (3- ((tert-butyldimethylsilyl) oxy) azetidin-1-yl) -2-oxoethyl) amino) pyrrolidin-1-yl) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To (R) -2- { [2- (3- ((2- (3- ((tert-butyldimethylsilyl) oxy) azetidin-1-yl) -2-oxoethyl) amino) pyrrolidin-1-yl) -6-ethylimidazo [2,1-b][1,3,4]Thiadiazol-5-yl]To a solution of (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (100mg, 0.14mmol) in acetonitrile were added triethylamine (40. mu.L, 0.28mmol) and dimethylcarbamoyl chloride (16. mu.L, 0.17mmol), and the resulting mixture was heated under reflux for 4 hours. After cooling to room temperature, 1.0N hydrochloric acid (1.0mL) was added to the reaction mixture, and the mixture was stirred for 30 minutes and concentrated under reduced pressure. Water (10mL) was then added to the residue, extracted with dichloromethane (15mL x 3), the organic layers were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the resulting residue was purified on a thick prep plate (dichloromethane: methanol ═ 16:1) to give the title compound as a white solid (20 mg).¹H NMR(400MHz, CDCl₃)δ:8.17(m,2H),7.18(m,2H),4.72-4.63(m,1H),4.48-4.37(m,2H),4.30-4.22(m,1H),4.09-4.01(m,1H), 3.94-3.86(m,1H),3.80-3.74(m,1H),3.72-3.66(m,2H),3.66-3.61(m,1H),3.60(s,3H),3.49-3.41(m,2H),2.87(s, 6H),2.64-2.58(m,2H),2.39-2.31(m,1H),2.26-2.19(m,1H),1.32-1.28(m,3H)。MS(m/z):653.1[M+1]。

Example 8

Preparation of cis-2- { [ 6-ethyl-2- (3-fluoro-4- ((2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) (methyl) amino) pyrrolidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

Step 1) cis-3- { [ (benzyloxy) carbonyl ] amino } -4-fluoropyrrolidine-1-carboxylic acid tert-butyl ester

To a solution of cis-3-amino-4-fluoropyrrolidine-1-carboxylic acid tert-butyl ester (0.3g, 1.47mmol) in dichloromethane (5mL) was added triethylamine (0.3mL, 2.14mmol), followed by cooling to 0 deg.C under nitrogen, and benzyl chloroformate (0.25mL, 1.76mmol) was added dropwise. The mixture was allowed to warm slowly to room temperature and stirred for 2 hours, after LC-MS showed completion of the reaction, the reaction was poured into water (15mL), extracted with ethyl acetate (15mL × 3), the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the resulting residue was purified by column chromatography (petroleum ether: ethyl acetate ═ 1:1) to give the title compound as a pale yellow oil (0.41g, 81%). MS (M/z):339.2[ M +1 ].

Step 2) cis- (4-fluoropyrrolidin-3-yl) carbamic acid benzyl ester

To a solution of cis-3- { [ (benzyloxy) carbonyl ] amino } -4-fluoropyrrolidine-1-carboxylic acid tert-butyl ester (0.41g, 1.21mmol) in dichloromethane (4mL) was added trifluoroacetic acid (1mL, 6 mmol). The resulting mixture was stirred at room temperature for 2 hours and after LC-MS showed the reaction was complete, it was concentrated under reduced pressure to give 0.28g of crude product, which was used in the next reaction without purification. MS (m/z): 239.2[ M +1 ].

Step 3) cis- [1- (5-acetylamino-6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl) -4-fluoropyrrolidin-3-yl ] carbamic acid benzyl ester

To a solution of benzyl cis- (4-fluoropyrrolidin-3-yl) carbamate (0.28g, 1.17mmol) in DMF (5mL) was added N- (2-bromo-6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl) carboxamide (0.32g, 1.17mmol) and potassium carbonate (0.48g, 3.47mmol), and the mixture was heated to 90 ℃ for overnight reaction. After the reaction solution was cooled to room temperature, the reaction solution was poured into water (15mL), extracted with ethyl acetate (15mL × 3), the organic layers were combined, washed with saturated brine (20mL × 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the resulting residue was purified by column chromatography (dichloromethane: methanol ═ 50:1) to obtain the objective product (85mg, 16%). MS (m/z): 432.2[ M +1 ].

Step 4) benzyl cis- {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl ] -4-fluoropyrrolidin-3-yl } (methyl) carbamate

To a solution of benzyl cis- [1- (5-acetylamino-6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl) -4-fluoropyrrolidin-3-yl ] carbamate (80mg, 0.18mmol) in THF (5mL) at 0 ℃ under nitrogen protection was added 60% sodium hydride (14.8mg, 0.36mmol), the mixture was stirred for 10 minutes, and then a solution of 2-chloro-4- (4-fluorophenyl) thiazole-5-carbonitrile (42.3mg, 0.18mmol) in tetrahydrofuran (3mL) was added dropwise. The mixture was allowed to warm to room temperature for 20 minutes, the reaction was monitored on a thin layer plate for completion, methyl iodide (55mg, 0.39mmol) was added, and the reaction was continued at room temperature for 2 hours. The reaction was checked by LC-MS for completion, water was added dropwise to quench the reaction, water (15mL) was added to the mixture, extraction was performed with ethyl acetate (15mL × 2), the combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the resulting residue was purified by column chromatography (petroleum ether: ethyl acetate ═ 2:1) to give the product (90mg, 79%). MS (m/z): 635.2[ M +1 ].

Step 5) cis-2- { [ 6-ethyl-2- (3-fluoro-4- (methylamino) pyrrolidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

Trimethyliodosilane (59. mu.L, 0.42mmol) was added dropwise to a solution of cis- {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl ] -4-fluoropyrrolidin-3-yl } (methyl) benzyl carbamate (90mg, 0.14mmol) in dichloromethane (4mL) at 0 ℃ under nitrogen. After the mixture was stirred at room temperature for 1 hour, water was added to quench the reaction, the reaction was concentrated under reduced pressure, water (15mL) and methyl tert-butyl ether (15mL) were added to the residue, the layers were separated, the aqueous phase was adjusted to pH 8 with sodium bicarbonate and extracted with ethyl acetate (15 mL. times.2), the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate and dried by spin-drying to give the desired product (60mg, 84%). MS (m/z): 501.1[ M +1 ].

Step 6) cis-2- { [ 6-ethyl-2- (3-fluoro-4- ((2- (3-hydroxyazetidin-1-yl) -2-oxoethyl) (methyl) amino) pyrrolidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To a mixture of cis-2- { [ 6-ethyl-2- (3-fluoro-4- (methylamino) pyrrolidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (60mg, 0.12mmol) in acetonitrile (3mL) was added 2-chloro-1- (3-hydroxyazetidin-1-yl) ethanone (17.6mg, 0.12mmol), potassium carbonate (33mg, 0.24mmol) and potassium iodide (10mg, 0.06mmol), and the mixture was heated to 80 ℃ for 2 hours. After the reaction solution was cooled to room temperature, filtered and concentrated under reduced pressure, the resulting residue was purified on thick prep plates (dichloromethane: methanol ═ 20:1) to give the title compound as a pale yellow solid (30mg, 41%). MS (m/z): 614.2[ M +1 ].

Example 9

Preparation of 2- { [ 6-Ethyl-2- (3- (2- (3-hydroxyazetidin-1-yl) -2-oxoethoxy) pyrrolidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

Step 1) (R) -2- { [2- (3-Aminopyrrolidin-1-yl) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To a solution of 2- { [ 2-bromo-6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (150mg, 0.32 mmol) in N, N-dimethylformamide (3.0mL) were added 3-pyrrolidinol (33mg, 0.39mmol) and potassium carbonate (89mg, 0.65mmol), and the mixture was reacted at room temperature for 5 hours. The reaction was poured into water (15mL), extracted with ethyl acetate (15mL x 2), the combined organic layers washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give a white solid, and slurried with petroleum ether to purify the title compound (150mg, 98%).

Step 1)2- { [ 6-Ethyl-2- (3- (2- (3-hydroxyazetidin-1-yl) -2-oxoethoxy) pyrrolidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To (R) -2- { [2- (3-aminopyrrolidin-1-yl) -6-ethylimidazo [2,1-b][1,3,4]Thiadiazol-5-yl]To a solution of (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (47mg, 0.10mmol) in tetrahydrofuran (3.0mL) was added 60% sodium hydride (20mg, 0.50mmol), the mixture was stirred at room temperature for 15 minutes, then 1- (3- ((tert-butyldimethylsilyl) oxy) azetidin-1-yl) -2-chloroacetophenone (33mg, 0.12mmol) was added, and the reaction was heated to 55 ℃ for 5 h. After the reaction mixture was cooled to 0 ℃, water was added dropwise to quench the reaction, and 1.0N hydrochloric acid (5.0mL) was added thereto and stirred at room temperature for 30 minutes. The mixture was concentrated under reduced pressure, water (15mL) was added and then extracted with dichloromethane (15mL x 3), the combined organic layers were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the resulting residue was purified on thick prep plates (dichloromethane: methanol ═ 30:1) to give the title compound as a white solid (18mg, 31%).¹H-NMR(400MHz,CDCl₃)δ:8.17(m,2H),7.18(m,2H), 4.72-4.65(m,1H),4.47-4.39(m,1H),4.33-4.26(m,2H),4.11-4.03(m,3H),3.95-3.88(m,1H),3.64-3.60(m,5H), 3.60-3.51(m,2H),2.64-2.59(m,2H),2.32-2.24(m,1H),2.21-2.11(m,1H),1.32-1.28(m,3H)。LC-MS(m/z):583.2 [M+1]。

Example 10

Preparation of (R) -2- { [ 6-ethyl-2- (3- (2- (3-hydroxyazetidin-1-yl) -2-oxoethoxy) pyrrolidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

Referring to example 9, (R) -2- { [ 6-ethyl-2- (3- (2- (3-hydroxyazetidin-1-yl) -2-oxoethoxy) pyrrolidin-1-yl) imidazo [2,1-b ] was prepared by substituting (R) -3-pyrrolidinol for 3-pyrrolidinol][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile.¹H-NMR(500MHz,CDCl₃)δ:8.17(m,2H),7.18(m,2H),4.73-4.66(m,1H),4.47-4.40(m,1H),4.34-4.27(m,2H), 4.12-4.08(m,1H),4.07(m,2H),3.95-3.88(m,1H),3.64-3.60(m,5H),3.60-3.52(m,2H),2.64-2.59(m,2H), 2.32-2.25(m,1H),2.20-2.11(m,1H),1.32-1.29(m,3H)。LC-MS(m/z):583.3[M+1]。

Example 11

Preparation of (S) -2- { [ 6-ethyl-2- (3- (2- (3-hydroxyazetidin-1-yl) -2-oxoethoxy) pyrrolidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

Referring to example 9, (S) -2- { [ 6-ethyl-2- (3- (2- (3-hydroxyazetidin-1-yl) -2-oxoethoxy) pyrrolidin-1-yl) imidazo [2,1-b ] was prepared by substituting 3-pyrrolidinol with (S) -3-pyrrolidinol][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile.¹H-NMR(500MHz,CDCl₃)δ:8.17(m,2H),7.18(m,2H),4.71-4.66(m,1H),4.46-4.39(m,1H),4.33-4.27(m,2H), 4.12-4.03(m,3H),3.95-3.88(m,1H),3.63-3.60(m,5H),3.60-3.52(m,2H),2.64-2.59(m,2H),2.32-2.24(m,1H), 2.20-2.12(m,1H),1.31-1.28(m,3H)。LC-MS(m/z):583.3[M+1]。

Example 12

Preparation of (R) -2- { [ 6-ethyl-2- (3- (2- (3-hydroxyazetidin-1-yl) -2-oxoethoxy) piperidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

Referring to example 9, (R) -2- { [ 6-ethyl-2- (3- (2- (3-hydroxyazetidin-1-yl) -2-oxoethoxy) piperidin-1-yl) imidazo [2,1-b ] was prepared using (R) -3-piperidinol instead of 3-pyrrolidinol][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile.¹H-NMR(500MHz,CDCl₃)δ:8.17(m,2H),7.18(m,2H),4.68-4.57(m,1H),4.46-4.37(m,1H),4.31-4.24(m,1H), 4.12-4.03(m,3H),3.92-3.84(m,1H),3.66-3.60(m,2H),3.60(m,3H),3.54-3.46(m,1H),3.42(m,2H),2.63-2.59(m, 2H),2.00-1.91(m,2H),1.83-1.77(m,1H),1.69-1.65(m,1H),1.32-1.29(m,3H)。LC-MS(m/z):597.2[M+1]。

Example 13

Preparation of (S) -2- { [ 6-ethyl-2- (3- (2- (3-hydroxyazetidin-1-yl) -2-oxoethoxy) piperidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

With reference to example 9, (S) -2- { [ 6-ethyl-2- (3- (2- (3-hydroxyazetidin-1-yl) -2-oxoethoxy) piperidin-1-yl) imidazo [2,1-b ] was prepared using (S) -3-piperidinol instead of 3-pyrrolidinol][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile.¹H-NMR(500MHz,CDCl₃)δ:8.17(m,2H),7.18(m,2H),4.68-4.58(m,1H),4.46-4.37(m,1H),4.31-4.25(m,1H), 4.12-4.04(m,3H),3.92-3.85(m,1H),3.67-3.61(m,2H),3.61(m,3H),3.55-3.47(m,1H),3.42(m,2H),2.63-2.59(m, 2H),1.99-1.91(m,2H),1.83-1.77(m,1H),1.68-1.63(m,1H),1.32-1.29(m,3H)。LC-MS(m/z):597.2[M+1]。

Example 14

Preparation of 2- { [ 6-Ethyl-2- (6- (3-hydroxyazetidine-1-carbonyl) -3-azabicyclo [3.1.0] hex-3-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

Step 1) tert-butyl 6- (3-hydroxyazetidine-1-carbonyl) -3-azabicyclo [3.1.0] hexane-3-carboxylate

To a solution of 3- (tert-butoxycarbonyl) -3-azabicyclo [3.1.0] hexane-6-carboxylic acid (30mg, 0.132mmol) in dichloromethane (10mL) was added HATU (75mg, 0.198mmol), azetidine-3-ol hydrochloride (17mg, 0.158mmol), and N, N-diisopropylethylamine (41mg, 0.317 mmol). The mixture was allowed to react at room temperature for 6 hours and the reaction was monitored by TLC for completion. Dichloromethane (5mL) and water (10mL) were added to the reaction mixture, stirred for 5 minutes, separated, and the organic phase was washed with 0.5M HCl (10mL) and saturated brine (10mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 43mg of a white solid, which was used in the next reaction without purification. LC-MS (M/z):283.2 [ M +1 ].

Step 2) 3-azabicyclo [3.1.0] hex-6-yl (3-azetidin-1-yl) methanone hydrochloride

35% ethanol hydrochloride solution (1mL) was added dropwise to a solution of crude tert-butyl 6- (3-hydroxyazetidine-1-carbonyl) -3-azabicyclo [3.1.0] hexane-3-carboxylate obtained in the previous step in dichloromethane (10mL) under ice-cooling, and the mixture was reacted at room temperature overnight. The reaction mixture was concentrated to give 51mg of a colorless oil, which was used as it was in the next reaction. LC-MS (M/z):183.1[ M +1 ].

Step 3)2- { [ 6-Ethyl-2- (6- (3-hydroxyazetidine-1-carbonyl) -3-azabicyclo [3.1.0] hex-3-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

To 3-azabicyclo [3.1.0]To a solution of hex-6-yl (3-azetidin-1-yl) methanone hydrochloride (51mg, 0.24mmol) in DMF (10mL) were added potassium carbonate (46mg, 0.33mmol) and 2- { [ 2-bromo-6-ethylimidazo [2,1-b ]][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile (61mg, 0.13mmol), and the mixture was reacted at 70 ℃ overnight. Filtration, concentration of the filtrate to dryness under reduced pressure from an oil pump, and purification of the residue using thick preparative plates (dichloromethane)Methanol 15:1) to give the title compound as a pale yellow solid (27mg, 36%).¹H NMR(400 MHz,CDCl₃)δ8.15(m,2H),7.16(m,2H),4.71(m,1H),4.44(m,1H),4.25(m,1H),4.08(m,1H),3.86(m,1H), 3.64(m,4H),3.57(s,3H),2.59(q,J＝7.6Hz,2H),2.26(m,2H),1.30(t,J＝7.6Hz,3H),0.77(m,2H)。MS(m/z)： 565.1[M+1]。

Example 15

Preparation of 2- { [ 6-Ethyl-2- (3- (3-hydroxyazetidine-1-carbonyl) -8-azabicyclo [3.2.1] octan-8-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

With reference to example 14, using 8- (tert-butyloxycarbonyl) -8-azabicyclo [3.2.1]Octane-3-carboxylic acid instead of 3- (tert-butyloxycarbonyl) -3-azabicyclo [3.1.0]Hexane-6-Carboxylic acid preparation to give 2- { [ 6-Ethyl-2- (3- (3-hydroxyazetidine-1-carbonyl) -8-azabicyclo [3.2.1]Octane-8-yl) imidazo [2,1-b][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile.¹H NMR(500MHz,CDCl₃)δ8.14(m,2H), 7.16(m,2H),4.68(m,1H),4.41-4.25(m,2H),4.24-4.09(m,2H),4.02(m,1H),3.83(m,1H),3.58(s,3H),2.78(m, 1H),2.59(q,J＝7.5Hz,2H),2.15(m,4H),1.80(m,2H),1.66(m,2H),1.30(t,J＝7.5Hz,3H)。MS(m/z)：593.2 [M+1]。

Example 16

Preparation of 2- { [ 6-Ethyl-2- (6- (3-hydroxyazetidine-1-carbonyl) -2-azaspiro [3.3] heptan-2-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

With reference to example 14, using 2- (tert-butyloxycarbonyl) -2-azaspiro [3.3]]Heptane-6-carboxylic acid instead of 3- (tert-butoxycarbonyl) -3-azabicyclo [3.1.0]Hexane-6-carboxylic acid preparation to give 2- { [ 6-ethyl-2- (6- (3-hydroxy)Azetidine-1-carbonyl) -2-azaspiro [3.3]Heptane-2-yl) imidazo [2,1-b][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile.¹H NMR(500MHz,CDCl₃)δ8.14(m,2H), 7.16(m,2H),4.67(m,1H),4.33-4.19(m,2H),4.09(m,4H),3.92(m,1H),3.85(m,1H),3.56(s,3H),2.91(m,1H), 2.63-2.49(m,4H),2.42(m,2H),1.29(t,J＝7.5Hz,3H)。MS(m/z)：579.1[M+1]。

Example 17

Preparation of 2- {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2,1-b ] [1,3,4] thiadiazol-2-yl ] pyrrolidin-3-yl } -N- (3-hydroxycyclobutyl) acetamide

Referring to example 14, 2- (1- (tert-Butoxycarbonyl) pyrrolidin-3-yl) acetic acid was used in place of 3- (tert-butyloxycarbonyl) -3-azabicyclo [ 3.1.0%]Hexane-6-carboxylic acid and 3-aminocyclobutanol hydrochloride instead of azetidine-3-ol hydrochloride gave 2- {1- [5- ((5-cyano-4- (4-fluorophenyl) thiazol-2-yl) (methyl) amino) -6-ethylimidazo [2, 1-b-][1,3,4]Thiadiazol-2-yl]Pyrrolidin-3-yl } -N- (3-hydroxycyclobutyl) acetamide.¹H NMR (400MHz,CDCl₃)δ8.16(m,2H),7.17(m,2H),5.69(m,1H),4.49(m,1H),3.95(m,1H),3.74(m,1H), 3.60-3-43(m,5H),3.15(m,1H),2.82(m,2H),2.61(q,J＝7.6Hz,2H),2.37-2.22(m,5H),1.84-1.76(m,3H),1.30(t, J＝7.6Hz,3H)。MS(m/z)：581.3[M+1]。

Example 18

Preparation of 2- { [ 6-Ethyl-2- (3- (3- (3-hydroxyazetidin-1-yl) -3-oxopropyl) pyrrolidin-1-yl) imidazo [2,1-b ] [1,3,4] thiadiazol-5-yl ] (methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile

With reference to example 14, 3- [1- (tert-butyloxycarbonyl) pyrrolidin-3-yl group]Propionic acid instead of 3- (tert-butoxy)Carbonyl) -3-azabicyclo [3.1.0]Hexane-6-carboxylic acid preparation to give 2- { [ 6-Ethyl-2- (3- (3- (3-hydroxyazetidin-1-yl) -3-oxopropyl) pyrrolidin-1-yl) imidazo [2,1-b][1,3,4]Thiadiazol-5-yl](methyl) amino } -4- (4-fluorophenyl) thiazole-5-carbonitrile.¹H NMR(400MHz,CDCl₃)δ8.16(m,2H),7.18(m,2H), 4.70(m,1H),4.34(m,1H),4.26(m,1H),4.00(m,1H),3.88(m,1H),3.66-3-52(m,5H),3.44(m,1H),3.09(m,1H), 2.61(q,J＝7.6Hz,2H),2.53(m,1H),2.41(m,1H),2.25-2.15(m,3H),1.85-1.72(m,3H),1.30(t,J＝7.6Hz,3H)。 MS(m/z)：581.3[M+1]。

Biological assay

In vitro analysis: enzyme activity screening using LPC as substrateThe principle is as follows: the lysophosphatidic acid (LPA) is hydrolyzed by lysoPLDase activity to generate lysophosphatidic acid (LPA) and choline, which is oxidized by choline oxidase to generate H₂O₂In the presence of Horse Radish Peroxidase (HRP), Amplex Red reagent and H₂O₂Reacting according to the chemical quantitative ratio of 1:1 to form a strong fluorescence product for fluorescence quantitative detection.

The experimental steps are as follows: test compounds were dissolved in DMSO as 10mM stock solutions and diluted in DMSO in a 3-fold gradient starting at 10mM concentration, 10 concentration points. A mixed solution 1 having a final concentration of 2 ng/. mu.l ATX, 2U/ml HRP and 0.2U/ml choline oxidase (choline oxidase) was prepared from the reaction buffer solution. Mu.l of the above mixed solution 1 was added to each well of the plate, and the DMSO-diluted compound was transferred into the plate at 10 nl/well using Echo 550. A mixed solution 2 was prepared in the reaction buffer solution to a final concentration of 60mM LPC and 400uM Amplex Red, and 20. mu.l of the mixed solution 2 was added to each well of the plate. After sample addition, the plate was shaken on a plate shaking instrument for 30s and incubated at room temperature for 30 min. Excitation light was read at 530nm using Envision and a fluorescence signal of 590nm was emitted. Calculating the inhibition rate of the compound on enzyme reaction according to the fluorescence ratio, and analyzing by using software to calculate the IC of the compound₅₀Values, see table 1.

TABLE 1 ATX inhibitory Activity assay of the Compounds of the invention based on LPC

+：≥1000nM；++：500-1000nM；+++：100-500nM；++++：0.01-100nM

Test compounds	LPC-IC₅₀	Test compounds	LPC-IC₅₀
				Example 1	++++	Example 2	++++
Example 3	+++	Example 4	++++
				Example 5	+++	Example 6	++++
Example 7	++++	Example 8	+++
				Example 9	++++	Example 10	+++
Example 11	++++	Example 13	++++
				Example 14	+++	Example 15	+
Example 16	+++	Example 17	++++
				Example 18	++++

Note: ND: no test was performed

As can be seen from Table 1, the compounds of the present invention have very good ATX inhibitory activity, and most of the compounds have IC₅₀Values were all below 100 nM.

In vitro analysis: screening for human plasma LPA detection activityThe principle is as follows: using LPC present in plasma as substrate, the percentage of residual activity at various test compound concentration points was determined by LC/MS/MS quantitative analysis of the resulting LPA18:2 (internal standard LPA17:0), and the IC was calculated from the ratio of the amount of LPA18:2 produced to the amount produced in the absence of test compound₅₀The value is obtained.

The experimental steps are as follows: human pooled plasma from at least 6 individuals, testedThe compound is diluted by 3 times of the diluent from the stock solution in a gradient manner to prepare working solutions (containing zero point) with 8 series of concentrations. 10 μ L of the thawed blank plasma sample was added to a solution of ice methanol containing an internal standard (LPA17:0) for direct protein precipitation as a system control sample. Respectively taking 2 mu L of working solution with serial concentrations, adding 198 mu L of human blank plasma, incubating at the concentration of 0-10 mu M, and putting the sample into a container containing 5% CO₂At 37 ℃ for 2 hours. After incubation, 10. mu.L of plasma sample was taken, an appropriate volume of ice-methanol solution containing an internal standard (LPA17:0) was added to precipitate proteins, the supernatant was centrifuged and subjected to LC/MS/MS for LPA18:2 detection, and analysis with software calculated the IC of the compound₅₀Values, see table 2.

Table 2 human whole blood analysis IC of compounds of the invention₅₀

+：≥1000nM；++：500-1000nM；+++：100-500nM；++++：0.01-100nM

Test compounds	Plasma active IC₅₀	Test compounds	Plasma active IC₅₀
				Example 1	+++	Example 2	++++
Example 3	ND	Example 4	++++
				Example 5	+++	Example 6	++++
Example 7	++++	Example 8	ND
				Example 9	ND	Example 10	ND
Example 11	++++	Example 12	ND
				Example 13	++++	Example 14	ND
Example 15	ND	Example 16	ND
				Example 17	++++	Example 18	++++

Note: ND: no test was performed

As shown in Table 2, the compounds of the present invention are also effective in inhibiting ATX in human plasma, thereby inhibiting LPC from being hydrolyzed to LPA, IC of the compounds₅₀Values were substantially below 100 nM.

Pharmacokinetic testing of rats

Male SD rat test animals were selected for this study, and plasma drug concentrations at different time points of intravenous injection and oral administration of the test compound of the present invention were quantitatively determined by LC/MS method in rats, respectively, to evaluate the pharmacokinetic profile of the test compound in SD rats.

A clear solution of the test compound of the present invention was injected into SD rats via the foot vein (free diet, 6-8 weeks old), and the suspension of the test compound was gavaged to SD rats (free diet, 6-8 weeks old). Animals collect blood samples in tail veins 0.083, 0.25, 0.5, 1,2, 4, 6, 8, 10 and 24 hours after administration, the blood collection amount is 0.15mL each time, all collected whole blood samples are placed in a centrifuge tube containing EDTA-K2, the centrifuge tube is turned upside down to fully mix the anticoagulant with the blood, the plasma is separated by centrifugation for 10min at 1500g and 4 ℃ within 30min, the plasma samples are transferred to a new centrifuge tube and stored at-90 to-60 ℃ for analysis. The plasma concentration of the test compound of the invention was measured by LC-MS/MS, pharmacokinetic parameters were calculated using a non-compartmental model in the Pharsight Phoenix 8.0 software, and the absolute bioavailability was calculated.

Pharmacokinetic studies show that the compound has better half-life and exposure in vivo and has good bioavailability.

In a word, the compound has good inhibitory activity on ATX, excellent in-vivo and in-vitro drug effect and pharmacokinetic property, and good clinical application prospect.

All publications, patents and patent applications cited in this specification are herein incorporated by reference as if each individual publication, patent or patent application were specifically and individually indicated to be incorporated by reference. Although the claimed subject matter has been described in terms of various examples/embodiments, those skilled in the art will recognize that various modifications/alterations, substitutions, deletions, and changes/variations may be made without departing from the spirit of the invention. Accordingly, it is intended that the scope of the claimed subject matter be limited only by the scope of the appended claims, including equivalents thereof.

Claims

1. A compound having the structure shown in formula (II):

wherein,

Cy²is that

Wherein the Cy is²Optionally substituted by 1,2,3 or 4R⁴Substitution;

Y²is- (CH)₂)_1-2-O-、-C(＝O)-CH₂-O-、-NHC(＝O)-CH₂-、-C(＝O)-(CH₂)₁-、-C(＝O)-CH₂-N(R^1b) -, or-NHC (═ O) NH-;

z is

R^1aIs H, C_1-4Alkyl, or halo C_1-4An alkyl group;

each R⁴Each independently is H, D, oxo, -CN, -NO₂、-OH、-NH₂、F、Cl、Br、I、C_1-4Alkyl, halo C_1-4Alkyl, cyano-substituted C_1-4Alkyl radical, C_1-4Hydroxyalkyl radical, C_1-4Alkoxy, halo C_1-4Alkoxy, or halo C_1-4Alkoxy radical C_1-4An alkyl group;

each R⁵Each independently is H, D, oxo, -CN, -NO₂、-OH、-NH₂、F、Cl、Br、I、C_1-4Alkyl, halo C_1-4Alkyl, cyano-substituted C_1-4Alkyl radical, C_1-4Hydroxyalkyl radical, C_1-4Alkoxy, halo C_1-4Alkoxy, or halo C_1-4Alkoxy radical C_1-4An alkyl group;

each R⁶Each independently is H, oxo, -CN, -NO₂、-OH、-NH₂、F、Cl、Br、I、C_1-4Alkyl, halo C_1-4Alkyl, cyano-substituted C_1-4Alkyl radical, C_1-4Hydroxyalkyl radical, C_1-4Alkoxy, halo C_1-4Alkoxy, or halo C_1-4Alkoxy radical C_1-4An alkyl group;

each R⁷Each independently is H, oxo, -CN, -NO₂、-OH、-NH₂、F、Cl、Br、I、C_1-4Alkyl, halo C_1-4Alkyl, cyano-substituted C_1-4Alkyl radical, C_1-4Hydroxyalkyl radical, C_1-4Alkoxy, halo C_1-4Alkoxy, or halo C_1-4Alkoxy radical C_1-4An alkyl group;

X⁶is N, orCH；

Each m4 is independently 1,2,3 or 4;

each m5 is independently 0, 1,2,3, or 4;

each m6 is independently 1,2,3 or 4;

n2 is 0, 1,2,3 or 4; and

t is 0, 1,2,3 or 4;

or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, having the structure of (IIa):

or a pharmaceutically acceptable salt thereof.

3. The compound of claim 1 or 2, wherein Z is

4. The compound of claim 1, wherein R²Is H, -CN, -NO₂、-OH、-NH₂F, Cl, Br, I, methyl, ethyl, propyl, isopropyl, butyl, hydroxymethyl, hydroxyethyl, trifluoromethyl, or trifluoroethyl.

5. The compound according to claim 1 or 2,wherein R is^1bIs CH₃-C(＝O)-、CH₃CH₂-C(＝O)-、CH₃-S(＝O)_1-2-、CH₃CH₂-S(＝O)_1-2-、(CH₃)₂N-C (═ O) -, or azetidinyl-C (═ O) -CH₂-, said R^1bOptionally substituted by 1,2,3 or 4R⁷Substitution; and

each R⁷Each independently is H, oxo, -CN, -NO₂、-OH、-NH₂F, Cl, Br, I, methyl, ethyl, propyl, isopropyl, tert-butyl, methoxy, ethoxy, -OCH₂CF₃、-CF₃、-CH₂CF₃、-CH₂CH₂CN、-CH₂CH₂OH, or CHF₂-O-CH₂-。

6. The compound of claim 1, wherein each R⁴、R⁵And R⁶Each independently is H, oxo, -CN, -NO₂、-OH、-NH₂F, Cl, Br, I, methyl, ethyl, propyl, isopropyl, tert-butyl, methoxy, ethoxy, -OCH₂CF₃、-CF₃、-CH₂CF₃、-CH₂CH₂CN、CHF₂-O-CH₂-、CF₃-O-CH₂-, or-CH₂CH₂OH。

7. A compound, which is a compound having one of the following structures:

or a pharmaceutically acceptable salt thereof.

8. A pharmaceutical composition comprising a compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable adjuvant, diluent or carrier.

9. The pharmaceutical composition of claim 8, further comprising an additional therapeutic agent.

10. Use of a compound according to any one of claims 1-7 or a pharmaceutical composition according to any one of claims 8-9 in the manufacture of a medicament for the prevention or treatment of a disease characterized by pathology with increased ATX expression in a mammal.

11. The use according to claim 10, wherein said diseases characterized by pathologies with increased ATX expression comprise: cancer, fibrotic disease, metabolic disease, myelodysplastic syndrome, cardiovascular disease, autoimmune disease, inflammation, neurological disease, or pain.

12. Use according to claim 10, wherein the disease with a pathological feature of increased ATX expression is pulmonary fibrosis or hepatic fibrosis.