A class of compounds for inducing cartilage formation and their applications

Abstract

The present invention discloses a class of small molecule compounds represented by structural formulas (I)-(VII) or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers, as well as pharmaceutical compositions containing such compounds, and their use in drugs for improving osteoarthritis or related diseases of articular cartilage injury by inducing the chondrogenic differentiation of endogenous mesenchymal stem cells, and their use in drugs for promoting the in vivo cartilage repair effect of stem cells by inducing the chondrogenic differentiation of mesenchymal stem cells in vitro to improve osteoarthritis or articular cartilage defects.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to a class of compounds for inducing cartilage formation and their applications. Background Art

[0002] Osteoarthritis (OA) is a slow degenerative disease characterized by the degradation and injury of articular cartilage, accompanied by hypertrophy of the joint margin and subchondral bone. Currently, there is a lack of safe and effective treatment methods for osteoarthritis in clinical practice. The current OA therapies mainly include oral administration of non-steroidal anti-inflammatory drugs (NSAIDs) or selective cyclooxygenase-2 (COX-2) inhibitors, intra-articular (IA) injection of agents such as corticosteroids and hyaluronic acid to relieve pain, and surgical treatments such as microfracture, joint replacement, and cartilage mosaicplasty. However, these treatment methods all have varying degrees of deficiencies and cannot repair irreversibly damaged cartilage, and most of the clinical results are not satisfactory. Therefore, new treatment means need to be developed to meet the increasingly severe clinical situation of OA.

[0003] Mesenchymal stem cells (MSCs) exist in the synovium, bone marrow, and adipose tissue of adults, as well as in neonatal cord blood. MSCs are pluripotent stem cells that can be isolated and proliferated in vitro. They can differentiate into several different types of cells and also have trilineage differentiation ability. Through the differentiation process, specialized cell types are formed from less specialized cell types, such as chondrocytes derived from MSCs. MSCs can differentiate into, including but not limited to, osteoblasts, chondrocytes, and adipocytes; they can also be induced to differentiate into chondrocytes in vitro. Cartistem is produced by Medipost in South Korea and was approved by the South Korean FDA on January 19, 2012. It is currently the only drug for the treatment of osteoarthritis with stem cells approved at the national level in the world. This drug is obtained by isolating stem cells from cord blood, inducing and culturing them, and then injecting the cultured cells into the joint cavity. The stem cells will differentiate into articular cartilage in the joint cavity, repair the articular cartilage injury, and thus fundamentally cure osteoarthritis. Since its launch, the sales of Cartistem have increased exponentially, demonstrating the broad market prospects for the stem cell treatment of OA.

[0004] Collagen is the main structural component of the epidermis. Collagen is very important for skin health and is widely used in the epidermal treatment of wrinkles and skin aging and can be used as a healing aid for burn patients. Collagen is generally expressed and produced in fibroblasts, and the collagen used for epidermal treatment or healing aid is generally human and bovine collagen.

[0005] However, there are still certain deficiencies in the treatment of OA by stem cell transplantation. Since OA is a heterogeneous disease with large individual differences and unstable clinical effects, at the same time, the in vivo differentiation effect of stem cell transplantation is low, and there is a risk of tumorigenicity in direct stem cell transplantation. Therefore, developing a class of drugs that can avoid the problems of stem cell drugs by inducing the differentiation of stem cells into chondrogenic progenitor cells in vitro and then transplanting them, or by injecting into the joint cavity to induce the differentiation of bone marrow mesenchymal stem cells (BMSCs) into chondrocytes. Small molecule drugs have the advantages of avoiding immune rejection, stability, low cost, and low cross-species risk compared with biological agents. Therefore, developing a class of drugs that can induce mesenchymal stem cells to become chondrocytes and improve diseases related to osteoarthritis or joint injuries is of great significance.

[0006] WO2011 / 008773 and WO2014 / 138687 describe a class of peptide compositions and their use for inducing the differentiation of mesenchymal cells into chondrocytes for the treatment or prevention of arthritis and joint injuries. In addition, WO2012 / 129562, WO2014 / 151953, WO2015 / 175487, and WO2018 / 225009 describe different small molecule compounds, compositions, and the use of those compositions in improving arthritis and joint injuries and inducing the differentiation of mesenchymal cells into chondrocytes. The compound kartogenin reported in Science in 2012 was the first small molecule drug with a clear mechanism that could induce mesenchymal stem cells to become chondrocytes, but due to its poor drug-likeness, it does not have the potential to be developed into a drug. Its subsequent developed drug KA34 was approved by the FDA to enter clinical phase I research in 2017. At the R&D Day event of Novartis in 2019, it was announced that its company's cartilage repair candidate drugs LNA043 and LRX712 had entered clinical phase I research, and RHH466 had entered the preclinical research stage. Generally speaking, using small molecules to induce the chondrogenic differentiation of stem cells to promote cartilage repair and treat osteoarthritis brings new hope for the treatment of this refractory disease OA.

[0007] Currently, drugs for arthritis are mainly used for relieving inflammation and pain and cannot repair cartilage damage. With the development of tissue regeneration technology, certain progress has been made in cartilage repair and other aspects. However, clinical studies have shown that the cartilage tissue repaired by the above methods is mostly fibrous cartilage tissue, rather than hyaline cartilage tissue that meets the mechanical requirements. Fortunately, research reports have shown that the cartilage tissue repaired by drug-induced chondrogenic differentiation of mesenchymal stem cells is hyaline cartilage, which can better mimic normal human cartilage in tissue repair, thus fundamentally repairing articular cartilage damage (Advanced Drug Delivery Reviews, 2019, 146: 289-305). However, currently, the research and development of drugs for cartilage repair are still scarce, and the exploration of compound structure diversity and the understanding of biological mechanisms are not sufficient. Therefore, the design and synthesis of novel and efficient small molecule inducers for inducing chondrogenic differentiation of MSCs and promoting cartilage repair have great research space and significance for the basic research and clinical application of OA treatment. Moreover, due to the continuous increase in the incidence of osteoarthritis, there is a continuous demand in society for effective bone and joint regeneration compositions and methods, which has a huge market prospect. Researchers are required to develop more compounds with high efficiency, low toxicity and good drug properties. Summary of the Invention

[0008] To solve the deficiencies of the existing technology, the object of the present invention is to provide a class of compounds or their pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers, or pharmaceutical compositions for treating and / or improving and / or preventing osteoarthritis or joint injuries, or for inducing the differentiation of mesenchymal stem cells into chondrocytes, and their applications in the preparation of drugs for treating and / or improving and / or preventing arthritis or articular cartilage injuries in mammals, and / or in the preparation of drugs for inducing the differentiation of mesenchymal stem cells into chondrocytes.

[0009] The present invention provides a class of compounds represented by the following formula (I) or their pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers, which are represented by the following structure (I):

[0010]

[0011] Wherein:

[0012] m is 0, 1, 2, 3 or 4; preferably, 0, 1;

[0013] n is 0, 1, 2, 3 or 4; preferably, 0, 1;

[0014] j is 0, 1, 2, 3 or 4; preferably, 1, 2;

[0015] k is 0, 1, 2, 3, 4 or 5; preferably, 0 or 1;

[0016] X is oxygen, C 1-6 alkyl, vinyl, ethynyl, -NR 3 R 4 -, -C(O)-, -NHC(O)-, -C(O)NH-, -NR 3 C(O)-, -C(O)NR 3 -, -NHC(O)NH-, -NHS(O)NH-, -SO 2 NH-, -NHSO 2 -, -N=N- etc.; preferably, -C(O)NH-;

[0017] (R 1 ) k refers to k substituents R attached to the A ring 1 , each independently selected from H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcyano, cyano, nitro, carboxyl, amino, azide, C 1-6 alkylhydroxy, OR 3 , SR 3 , S(O)R 3 , SO 2 R 3 , NR 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , -NR 3 C(O)R 4 , -C(O)NR 3 R 4 , C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl etc.; preferably, H, halogen, nitro, carboxyl, amino, -NR 3 C(O)R 4 , -C(O)NR 3 R 4 .

[0018] Among them, the heterocycloalkyl group is selected from C cycloalkyl groups containing 1-5 nitrogen, oxygen, or sulfur atoms; the aryl group is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; preferably, it is a benzene ring; among them, the cycloalkyl group, heterocycloalkyl group, aryl group, and heteroaryl group can be further substituted by 1-2 R 3-8 groups; 3

[0019] (R 2 ) j refers to j substituents R 2 connected to the B ring, each independently selected from H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcyano, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, cyano, nitro, carboxyl, amino, SR 3 , S(O)R 3 , SO 2 R 3 , N R 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , C(O)R 3 , C(O)NR 3 R 4 , C 1-6 alkyloxy, (CR 5 R 6 )OR 3 , (CR 5 R 6 )NR 3 R 4 , (CR 5 R​6 )(CR 7 R 8 )OR 3 、Y(CR 5 R 6 )(CR 7 R 8 )OR 3 、Y(CR 5 R 6 )(CR 7 R 8 )NR 3 R 4 、(CR 5 R 6 )C(O)R 3 、(CR 5 R 6 )C(O)OR 3 、(CR 5 R 6 )C(O)NR 3 R 4 、Y(CR 5 R 6 )C(O)R 3 、Y(CR 5 R 6 )C(O)OR 3 、Y(CR 5 R 6 )C(O)NR 3 R 4 、Y(CR 5 R 6 )(CR 7 R 8 )C(O)R 3 、Y(CR 5 R 6 )(CR 7 R 8 )C(O)OR 3 、Y(CR 5 R 6 )(CR 7 R 8 )C(O)NR 3 R 4 、(CR 5 R 6 )NR 3 SO 2 R 3 or C(=NOR 3 )R 3 etc.; preferably, it is aryl, C 1-6 alkyloxy. Wherein, the heterocyclic alkyl includes C containing 1-5 nitrogen, oxygen, or sulfur atoms3-8 Cycloalkyl; the aryl group is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; preferably, it is a benzene ring, C 1-6 alkoxy, Y(CR 5 R 6 )(CR 7 R 8 )OR 3 ; wherein, the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups can be further substituted by 1-2 R 3 groups;

[0020] Y is O or CR 9 R 10 ;

[0021] R 3 is selected from H, C 1-6 alkyl, C 1-6 alkylNHC(O)R 4 , C 1-6 alkylC(O)NHR 4 , p(C 2 alkyloxy)C 2 alkylNHC(O)R 4 , p(C 2 alkyloxy)C 2 alkylC(O)NHR 4 etc.; preferably, it is H, C 1-6 alkylC(O)NHR 4 ; where p is 0, 1, 2, 3, 4;

[0022] R 4 is H, C 1-6 alkyl, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, etc.; wherein, the heterocycloalkyl is selected from C containing 1-5 nitrogen, oxygen, or sulfur atoms 3-8Cycloalkyl; the aryl group is selected from phenyl, anthracenyl, naphthyl, phenanthryl, etc.; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuryl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl, heterocycloalkyl, aryl, heteroaryl group may be further substituted by 1-2 R 3 groups;

[0023] R 5 、R 6 、R 7 、R 8 、R 9 and R 10 are independently selected from H, halogen, C 1-6 alkyl, OH, CO 2 R 3 、NR 3 R 4 、C 1-6 alkyloxy, etc.; preferably, it is H;

[0024] Ring A and Ring B are each independently selected from any one of the following groups: C 3-8 cycloalkyl, C 7 -C 10 bridged ring, heterocycloalkyl containing one or more O, N, S atoms, aryl, five- or six-membered heteroaryl containing one or more O, N, S atoms, benzoheterocycle containing one or more O, N, S atoms; wherein, the heterocycloalkyl is selected from C 3 - 8 cycloalkyl containing 1-5 nitrogen, oxygen, sulfur atoms; the aryl group is selected from phenyl, anthracenyl, naphthyl, phenanthryl; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuryl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl; the benzoheterocycle is selected from benzopyrazolyl, benzopyranyl, pyridazinyl, benzopyrimidinyl, benzimidazolyl, benzothiazolyl, benzisothiazolyl, benzopiperazinyl, indolyl, quinolinyl, pyrazinyl, isoquinolinyl, benzoxazolyl, benzisoxazolyl, benzofuryl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl.

[0025] In the above formula (I), when Ring B is a benzene ring, it is represented by the following structure (II):

[0026]

[0027] Wherein:

[0028] m is 0, 1, 2, 3 or 4; preferably, 0 or 1;

[0029] n is 0, 1, 2, 3 or 4; preferably, 0 or 1;

[0030] j is 0, 1, 2, 3 or 4; preferably, 1 or 2;

[0031] k is 0, 1, 2, 3, 4 or 5; preferably, 0 or 1;

[0032] X is oxygen, C 1-6 alkyl, vinyl, ethynyl, -NR 3 R 4 -, -C(O)-, -NHC(O)-, -C(O)NH-, -NR 3 C(O)-, -C(O)NR 3 -, -NHC(O)NH-, -NHS(O)NH-, -SO 2 NH-, -NHSO 2 -, -N=N-, etc.; preferably, -C(O)NH-;

[0033] (R 1 ) k refers to k substituents R attached to the A ring 1 , each independently being H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcyano, cyano, nitro, carboxyl, amino, azide, C 1-6 alkylhydroxy, OR 3 , SR 3 , S(O)R 3 , SO 2 R 3 , NR 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , -NR 3 C(O)R 4 , -C(O)NR 3 R 4 , C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C1-6 Alkyl - heterocycloalkyl, C 1-6 Alkyl - aryl, C 1-6 Alkyl - O - aryl, C 1-6 Alkyl - heteroaryl, heteroaryl - aryl, etc.; preferably, H, halogen, nitro, carboxyl, amino, -NR 3 C(O)R 4 、-C(O)NR 3 R 4 。

[0034] Wherein, the heterocycloalkyl includes C 3-8 cycloalkyl containing 1 - 5 nitrogen, oxygen, or sulfur atoms; the aryl is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl, heterocycloalkyl, aryl, heteroaryl may be further substituted by 1 - 2 R 3 groups;

[0035] (R 2 ) j refers to j substituents R 2 connected to the benzene ring, each independently selected from H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkyl cyano, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl - cycloalkyl, C 1-6 alkyl - heterocycloalkyl, C 1-6 alkyl - aryl, C 1-6 alkyl - O - aryl, C 1-6 alkyl - heteroaryl, heteroaryl - aryl, cyano, nitro, carboxyl, amino, SR 3 、S(O)R 3 、SO 2 R 3 、N R 3 R 4 、COR 3 、C(O)R 3 、CO 2 R 3 、C(O)R 3 、CO2R3 、 C(O)NR 3 R 4 、 C 1-6 alkyloxy, (CR 5 R 6 )OR 3 、 (CR 5 R 6 )NR 3 R 4 、 (CR 5 R 6 )(CR 7 R 8 )OR 3 、 Y(CR 5 R 6 )(CR 7 R 8 )OR 3 、 Y(CR 5 R 6 )(CR 7 R 8 )NR 3 R 4 、 (CR 5 R 6 )C(O)R 3 、 (CR 5 R 6 )C(O)OR 3 、 (CR 5 R 6 )C(O)NR 3 R 4 、 Y(CR 5 R 6 )C(O)R 3 、 Y(CR 5 R 6 )C(O)OR 3 、 Y(CR 5 R 6 )C(O)NR 3 R 4 、 Y(CR 5 R 6 )(CR 7 R 8 )C(O)R 3 、 Y(CR 5 R 6 )(CR 7 R 8 )C(O)OR 3 、 Y(CR 5 R 6 )(CR 7 R 8 )C(O)NR3 R 4 、 (CR 5 R 6 )NR 3 SO 2 R 3 or C(=NOR 3 )R 3 etc.; preferably, it is a benzene ring, C 1-6 alkoxy, Y(CR 5 R 6 )(CR 7 R 8 )OR 3 ; wherein, the heterocycloalkyl group includes a C 3-8 cycloalkyl group containing 1-5 nitrogen, oxygen, or sulfur atoms; the aryl group is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuryl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl group, heterocycloalkyl group, aryl group, and heteroaryl group may be further substituted by 1-2 R 3 groups;

[0036] Y is O or CR 9 R 10 ;

[0037] R 3 is selected from H, C 1-6 alkyl, C 1-6 alkylNHC(O)R 4 , C 1-6 alkylC(O)NHR 4 , p(C 2 alkyloxy)C 2 alkylNHC(O)R 4 , p(C 2 alkyloxy)C 2 alkylC(O)NHR 4 etc.; preferably, it is H, C 1-6 alkylC(O)NHR 4 ; wherein p is 0, 1, 2, 3, 4;

[0038] R 4 is H, C 1-6 alkyl, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, etc.; wherein, the heterocycloalkyl includes C 3-8 cycloalkyl containing 1-5 nitrogen, oxygen, or sulfur atoms; the aryl includes phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl includes pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may be further substituted by 1-2 R 3 groups;

[0039] R 5 、R 6 、R 7 、R 8 、R 9 and R 10 are independently selected from H, halogen, C 1-6 alkyl, OH, CO 2 R 3 、NR 3 R 4 、C 1-6 alkyloxy, etc.; preferably, H;

[0040] Ring A is selected from any one of the following groups: C 3-8 cycloalkyl, C 7 -C 10 bridged ring, heterocycloalkyl containing one or more O, N, or S atoms, aryl, five- or six-membered heteroaryl containing one or more O, N, or S atoms, benzoheterocycle containing one or more O, N, or S atoms; wherein, the heterocycloalkyl is selected from C 3-8 cycloalkyl containing 1-5 nitrogen, oxygen, or sulfur atoms; the aryl is selected from phenyl, anthryl, naphthyl, phenanthryl; the heteroaryl is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl; the benzoheterocycle is selected from benzopyrazolyl, benzopyranyl, pyridazinyl, benzopyrimidinyl, benzimidazolyl, benzothiazolyl, benzisothiazolyl, benzopiperazinyl, indolyl, quinolinyl, pyrazinyl, isoquinolinyl, benzoxazolyl, benzisoxazolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl.

[0041] In the above formula (II), when X is -C(O)NH-, it is represented by the following structure (III):

[0042]

[0043] Wherein:

[0044] m is 0, 1, 2, 3 or 4; preferably, it is 0, 1;

[0045] n is 0, 1, 2, 3 or 4; preferably, it is 0, 1;

[0046] j is 0, 1, 2, 3 or 4; preferably, it is 1, 2;

[0047] k is 0, 1, 2, 3, 4 or 5; preferably, it is 0, 1;

[0048] (R 1 ) k refers to k substituents R attached to the A ring 1 , each independently being H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcyano, cyano, nitro, carboxyl, amino, azide, C 1-6 alkylhydroxy, OR 3 , SR 3 , S(O)R 3 , SO 2 R 3 , NR 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , -NR 3 C(O)R 4 , -C(O)NR 3 R 4 , C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, etc.; preferably, it is H, halogen, nitro, carboxyl, amino, -NR 3 C(O)R4 , -C(O)NR 3 R 4 .

[0049] Among them, the heterocycloalkyl group includes a C cycloalkyl group containing 1 - 5 nitrogen, oxygen, or sulfur atoms; the aryl group is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; among them, the cycloalkyl group, heterocycloalkyl group, aryl group, and heteroaryl group can be further substituted by 1 - 2 R 3-8 groups; 3

[0050] (R 2 ) j refers to j substituents R attached to the benzene ring 2 , each independently selected from H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkyl cyano, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl - cycloalkyl, C 1-6 alkyl - heterocycloalkyl, C 1-6 alkyl - aryl, C 1-6 alkyl - O - aryl, C 1-6 alkyl - heteroaryl, heteroaryl - aryl, cyano, nitro, carboxyl, amino, SR 3 , S(O)R 3 , SO 2 R 3 , N R 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , C(O)R 3 , CO2R 3 , C(O)NR 3 R 4 , C 1-6 alkyloxy, (CR 5 R 6 )OR 3 , (CR 5 R​6 )NR 3 R 4 、(CR 5 R 6 )(CR 7 R 8 )OR 3 、Y(CR 5 R 6 )(CR 7 R 8 )OR 3 、Y(CR 5 R 6 )(CR 7 R 8 )NR 3 R 4 、(CR 5 R 6 )C(O)R 3 、(CR 5 R 6 )C(O)OR 3 、(CR 5 R 6 )C(O)NR 3 R 4 、Y(CR 5 R 6 )C(O)R 3 、Y(CR 5 R 6 )C(O)OR 3 、Y(CR 5 R 6 )C(O)NR 3 R 4 、Y(CR 5 R 6 )(CR 7 R 8 )C(O)R 3 、Y(CR 5 R 6 )(CR 7 R 8 )C(O)OR 3 、Y(CR 5 R 6 )(CR 7 R 8 )C(O)NR 3 R 4 、(CR 5 R 6 )NR 3 SO 2 R 3 或C(=NOR 3 )R 3etc.; preferably, it is a benzene ring, C 1-6 alkoxy, Y(CR 5 R 6 )(CR 7 R 8 )OR 3 ; wherein, the heterocycloalkyl group includes a C 3-8 cycloalkyl group containing 1-5 nitrogen, oxygen, or sulfur atoms; the aryl group is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuryl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl group, heterocycloalkyl group, aryl group, and heteroaryl group may be further substituted by 1-2 R 3 groups;

[0051] Y is O or CR 9 R 10 ;

[0052] R 3 is selected from H, C 1-6 alkyl, C 1-6 alkylNHC(O)R 4 , C 1-6 alkylC(O)NHR 4 , p(C 2 alkyloxy)C 2 alkylNHC(O)R 4 , p(C 2 alkyloxy)C 2 alkylC(O)NHR 4 etc.; preferably, it is H, C 1-6 alkylC(O)NHR 4 ; where p is 0, 1, 2, 3, 4;

[0053] R 4 is H, C 1-6 alkyl, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, etc.; wherein, the heterocycloalkyl group includes a C 3-8Cycloalkyl; the aryl group includes phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group includes pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; among them, the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups may be further substituted by 1-2 R 3 groups;

[0054] R 5 、R 6 、R 7 、R 8 、R 9 and R 10 are independently selected from H, halogen, C 1-6 alkyl, OH, CO 2 R 3 、NR 3 R 4 、C 1-6 alkyloxy, etc.; preferably, it is H;

[0055] Ring A is selected from any one of the following groups: C 3-8 cycloalkyl, C 7 -C 10 bridged ring, heterocycloalkyl containing one or more O, N, S atoms, aryl, five- or six-membered heteroaryl containing one or more O, N, S atoms, benzoheterocycle containing one or more O, N, S atoms; among them, the heterocycloalkyl is selected from C 3-8 cycloalkyl containing 1-5 nitrogen, oxygen, sulfur atoms; the aryl is selected from phenyl, anthryl, naphthyl, phenanthryl; the heteroaryl is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl; the benzoheterocycle is selected from benzopyrazolyl, benzopyranyl, pyridazinyl, benzopyrimidinyl, benzimidazolyl, benzothiazolyl, benzisothiazolyl, benzopiperazinyl, indolyl, quinolinyl, pyrazinyl, isoquinolinyl, benzoxazolyl, benzisoxazolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl.

[0056] In the above formula (III), when m is 0 and n is 0, it is represented by the following structure (IV):

[0057]

[0058] Wherein:

[0059] j is 0, 1, 2, 3 or 4; preferably, 1, 2;

[0060] k is 0, 1, 2, 3, 4 or 5; preferably, 0, 1;

[0061] (R 1 ) k refers to k substituents R attached to the A ring 1 , each independently being H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcyano, cyano, nitro, carboxyl, amino, azide, C 1-6 alkylhydroxy, OR 3 , SR 3 , S(O)R 3 , SO 2 R 3 , NR 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , -NR 3 C(O)R 4 , -C(O)NR 3 R 4 , C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, etc.; preferably, H, halogen, nitro, carboxyl, amino, -NR 3 C(O)R 4 , -C(O)NR 3 R 4 .

[0062] Wherein, the heterocycloalkyl includes C containing 1-5 nitrogen, oxygen, sulfur atoms 3-8Cycloalkyl; the aryl group is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups may be further substituted by 1 to 2 R 3 groups;

[0063] (R 2 ) j refers to j substituents R 2 connected to the benzene ring, each independently selected from H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcyano, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, cyano, nitro, carboxyl, amino, SR 3 , S(O)R 3 , SO 2 R 3 , N R 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , C(O)R 3 , CO2R 3 , C(O)NR 3 R 4 , C 1-6 alkyloxy, (CR 5 R 6 )OR 3 , (CR 5 R 6 )NR 3 R 4 , (CR 5 R 6 )(CR 7 R 8 )OR3 , Y(CR 5 R 6 )(CR 7 R 8 ) OR 3 , Y(CR 5 R 6 )(CR 7 R 8 ) NR 3 R 4 , (CR 5 R 6 ) C(O)R 3 , (CR 5 R 6 ) C(O)OR 3 , (CR 5 R 6 ) C(O)NR 3 R 4 , Y(CR 5 R 6 ) C(O)R 3 , Y(CR 5 R 6 ) C(O)OR 3 , Y(CR 5 R 6 ) C(O)NR 3 R 4 , Y(CR 5 R 6 )(CR 7 R 8 ) C(O)R 3 , Y(CR 5 R 6 )(CR 7 R 8 ) C(O)OR 3 , Y(CR 5 R 6 )(CR 7 R 8 ) C(O)NR 3 R 4 , (CR 5 R 6 ) NR 3 SO 2 R 3 or C(=NOR 3 ) R 3 etc.; preferably, it is a benzene ring, C 1-6 alkoxy, Y(CR 5 R 6 )(CR 7 R 8 ) OR 3; wherein, the heterocycloalkyl group includes a C cycloalkyl group containing 1-5 nitrogen, oxygen, or sulfur atoms; the aryl group is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl group, heterocycloalkyl group, aryl group, and heteroaryl group may be further substituted by 1-2 R groups 3-8 ; 3

[0064] Y is O or CR 9 R 10 ;

[0065] R 3 is selected from H, C 1-6 alkyl, C 1-6 alkylNHC(O)R 4 、C 1-6 alkylC(O)NHR 4 、p(C 2 alkyloxy)C 2 alkylNHC(O)R 4 、p(C 2 alkyloxy)C 2 alkylC(O)NHR 4 etc.; preferably, it is H, C 1-6 alkylC(O)NHR 4 ; where p is 0, 1, 2, 3, 4;

[0066] R 4 is H, C 1-6 alkyl, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, etc.; wherein, the heterocycloalkyl group includes a C 3-8 ​Cycloalkyl; the aryl group includes phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group includes pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl, heterocycloalkyl, aryl, heteroaryl group can be further substituted by 1-2 R 3 groups;

[0067] R 5 、R 6 、R 7 、R 8 、R 9 and R 10 are independently selected from H, halogen, C 1-6 alkyl, OH, CO 2 R 3 、NR 3 R 4 、C 1-6 alkyloxy, etc.; preferably, it is H;

[0068] Ring A is selected from any one of the following groups: C 3-7 cycloalkyl, C 7 -C 10 bridged ring, heterocycloalkyl containing one or more O, N, S atoms, aryl, five- or six-membered heteroaryl containing one or more O, N, S atoms, benzoheterocycle containing one or more O, N, S atoms; wherein, the heterocycloalkyl is selected from C 3-8 cycloalkyl containing 1-5 nitrogen, oxygen, sulfur atoms; the aryl is selected from phenyl, anthryl, naphthyl, phenanthryl; the heteroaryl is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl; the benzoheterocycle is selected from benzopyrazolyl, benzopyranyl, pyridazinyl, benzopyrimidinyl, benzimidazolyl, benzothiazolyl, benzisothiazolyl, benzopiperazinyl, indolyl, quinolinyl, pyrazinyl, isoquinolinyl, benzoxazolyl, benzisoxazolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl.

[0069] In the above formula (IV), when Ring A is a thiophene ring, it is represented by the following structure (V):

[0070]

[0071] Wherein:

[0072] j is 0, 1, 2, 3 or 4, preferably 1 or 2;

[0073] k is 0, 1, 2 or 3, preferably 0, 1 or 2;

[0074] (R 1 ) k refers to k substituents R attached to the thiophene ring 1 , each independently being H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcyano, cyano, nitro, carboxyl, amino, azide, C 1-6 alkylhydroxy, OR 3 , SR 3 , S(O)R 3 , SO 2 R 3 , NR 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , -NR 3 C(O)R 4 , -C(O)NR 3 R 4 , C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, etc.; preferably H, halogen, nitro, carboxyl, amino, -NR 3 C(O)R 4 , -C(O)NR 3 R 4 .

[0075] Wherein, the heterocycloalkyl includes C containing 1-5 nitrogen, oxygen or sulfur atoms 3-8Cycloalkyl; the aryl group is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups may be further substituted by 1-2 R 3 groups;

[0076] (R 2 ) j refers to j substituents R 2 connected to the benzene ring, each independently selected from H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcyano, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, cyano, nitro, carboxyl, amino, SR 3 , S(O)R 3 , SO 2 R 3 , N R 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , C(O)R 3 , CO2R 3 , C(O)NR 3 R 4 , C 1-6 alkyloxy, (CR 5 R 6 )OR 3 , (CR 5 R 6 )NR 3 R 4 , (CR 5 R 6 )(CR 7 R 8 )OR3 , Y(CR 5 R 6 )(CR 7 R 8 )OR 3 , Y(CR 5 R 6 )(CR 7 R 8 )NR 3 R 4 , (CR 5 R 6 )C(O)R 3 , (CR 5 R 6 )C(O)OR 3 , (CR 5 R 6 )C(O)NR 3 R 4 , Y(CR 5 R 6 )C(O)R 3 , Y(CR 5 R 6 )C(O)OR 3 , Y(CR 5 R 6 )C(O)NR 3 R 4 , Y(CR 5 R 6 )(CR 7 R 8 )C(O)R 3 , Y(CR 5 R 6 )(CR 7 R 8 )C(O)OR 3 , Y(CR 5 R 6 )(CR 7 R 8 )C(O)NR 3 R 4 , (CR 5 R 6 )NR 3 SO 2 R 3 or C(=NOR 3 )R 3 etc.; preferably, it is a benzene ring, C 1-6 alkoxy, Y(CR 5 R 6 )(CR 7 R 8 )OR 3; wherein, the heterocycloalkyl group includes a C cycloalkyl group containing 1-5 nitrogen, oxygen, or sulfur atoms; 3-8 The aryl group is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl group, heterocycloalkyl group, aryl group, and heteroaryl group may be further substituted by 1-2 R 3 groups;

[0077] Y is O or CR 9 R 10 ;

[0078] R 3 is selected from H, C 1-6 alkyl, C 1-6 alkylNHC(O)R 4 , C 1-6 alkylC(O)NHR 4 , p(C 2 alkyloxy)C 2 alkylNHC(O)R 4 , p(C 2 alkyloxy)C 2 alkylC(O)NHR 4 etc.; preferably, it is H, C 1-6 alkylC(O)NHR 4 ; where p is 0, 1, 2, 3, 4;

[0079] R 4 is H, C 1-6 alkyl, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, etc.; wherein, the heterocycloalkyl group includes a C 3-8Cycloalkyl; the aryl group includes phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group includes pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups may be further substituted by 1-2 R 3 groups;

[0080] R 5 、R 6 、R 7 、R 8 、R 9 and R 10 are independently selected from H, halogen, C 1-6 alkyl, OH, CO 2 R 3 、NR 3 R 4 、C 1-6 alkyloxy, etc.; preferably, it is H;

[0081] In the above formula (V), when R 1 is connected to the thiophene ring through the W group, it is represented by the following structure (VI):

[0082]

[0083] Wherein:

[0084] j is 0, 1, 2, 3 or 4, preferably 1, 2;

[0085] W is oxygen, C 1-6 alkyl, vinyl, ethynyl, -NR 3 R 4 -, -C(O)-, -NHC(O)-, -C(O)NH-, -NR 3 C(O)-, -C(O)NR 3 -, -NHC(O)NH-, -NHS(O)NH-, -SO 2 NH-, -NHSO 2 -, -N=N-, etc.; preferably, it is -NHC(O)-, -C(O)NH-;

[0086] R 1 is H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkyloxy, C 1-6 haloalkyl, C1-6 Alkyl cyano, cyano, nitro, carboxyl, amino, azide, C 1-6 alkyl hydroxyl, OR 3 , SR 3 , S(O)R 3 , SO 2 R 3 , NR 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , -NR 3 C(O)R 4 , -C(O)NR 3 R 4 , C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-C(O)NR 3 R 4 , C 1-6 alkyl--NR 3 C(O)R 4 , C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, etc.; preferably, C 1-6 alkyl hydroxyl, C 1-6 alkyl-C(O)NR 3 R 4 , C 1-6 alkyl--NR 3 C(O)R 4 . Among them, the heterocycloalkyl includes C 3-8 cycloalkyl containing 1-5 nitrogen, oxygen, or sulfur atoms; the aryl is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; preferably, C 1-6 alkyl hydroxyl; the heteroaryl is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; among them, the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl may be further substituted by 1-2 R 3 groups;

[0087] (R 2 ) jThe j substituents R attached to the benzene ring 2 , each independently selected from H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkyl cyano, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, cyano, nitro, carboxyl, amino, SR 3 , S(O)R 3 , SO 2 R 3 , N R 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , C(O)R 3 , CO2R 3 , C(O)NR 3 R 4 , C 1-6 alkyloxy, (CR 5 R 6 )OR 3 , (CR 5 R 6 )NR 3 R 4 , (CR 5 R 6 )(CR 7 R 8 )OR 3 , Y(CR 5 R 6 )(CR 7 R 8 )OR 3 , Y(CR 5 R 6 )(CR 7 R 8 )NR 3 R 4 , (CR 5 R 6 )C(O)R 3 , (CR 5 R6 )C(O)OR 3 , (CR 5 R 6 )C(O)NR 3 R 4 、Y(CR 5 R 6 )C(O)R 3 、Y(CR 5 R 6 )C(O)OR 3 、Y(CR 5 R 6 )C(O)NR 3 R 4 、Y(CR 5 R 6 )(CR 7 R 8 )C(O)R 3 、Y(CR 5 R 6 )(CR 7 R 8 )C(O)OR 3 、Y(CR 5 R 6 )(CR 7 R 8 )C(O)NR 3 R 4 , (CR 5 R 6 )NR 3 SO 2 R 3 or C(=NOR 3 )R 3 etc.; preferably, a benzene ring, C 1-6 Alkoxy, Y(CR 5 R 6 )(CR 7 R 8 )OR 3 ; wherein the heterocycloalkyl group includes C containing 1-5 nitrogen, oxygen, and sulfur atoms 3-8 Cycloalkyl; the aryl is selected from phenyl, anthracenyl, naphthyl, phenanthryl, etc.; the heterocyclic aryl is selected from pyridyl, furanyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolyl, pyrazinyl, isoquinolyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazine, phenothiazinyl, pteridinyl, etc.; wherein the cycloalkyl, heterocycloalkyl, aryl, heterocyclic aryl may be substituted by 1-2 R 3 The group is further substituted; Y is O or CR 9 R10 ;

[0088] R 3 is selected from H, C 1-6 alkyl, C 1-6 alkyl NHC(O)R 4 , C 1-6 alkyl C(O)NHR 4 , p(C 2 alkyloxy)C 2 alkyl NHC(O)R 4 , p(C 2 alkyloxy)C 2 alkyl C(O)NHR 4 etc.; preferably, it is H, C 1-6 alkyl C(O)NHR 4 ; where p is 0, 1, 2, 3, 4;

[0089] R 4 is H, C 1-6 alkyl, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, etc.; where the heterocycloalkyl includes C 3-8 cycloalkyl containing 1-5 nitrogen, oxygen, or sulfur atoms; the aryl includes phenyl, anthracenyl, naphthyl, phenanthryl, etc.; the heteroaryl includes pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuryl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; where the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl can be further substituted by 1-2 R 3 groups;

[0090] R 5 , R 6 , R 7 , R 8 , R 9 and R 10 are independently selected from H, halogen, C 1-6 alkyl, OH, CO 2 R 3 , NR 3 R 4 , C 1-6Alkoxy group, etc.; preferably, it is H;

[0091] In the above formula (V), when W is 6-aminohexanamide, it is represented by the following structure (VII):

[0092]

[0093] Wherein:

[0094] j is 0, 1, 2, 3 or 4, preferably 1 or 2;

[0095] (R 2 ) j refers to j substituents R connected to the benzene ring 2 , each independently selected from H, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 alkylcyano, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, cyano, nitro, carboxyl, amino, SR 3 , S(O)R 3 , SO 2 R 3 , N R 3 R 4 , COR 3 , C(O)R 3 , CO 2 R 3 , C(O)R 3 , CO2R 3 , C(O)NR 3 R 4 , C 1-6 alkoxy, (CR 5 R 6 )OR 3 , (CR 5 R 6 )NR 3 R 4 , (CR 5 R 6 )(CR 7 R 8 )OR3 , Y(CR 5 R 6 )(CR 7 R 8 )OR 3 , Y(CR 5 R 6 )(CR 7 R 8 )NR 3 R 4 , (CR 5 R 6 )C(O)R 3 , (CR 5 R 6 )C(O)OR 3 , (CR 5 R 6 )C(O)NR 3 R 4 , Y(CR 5 R 6 )C(O)R 3 , Y(CR 5 R 6 )C(O)OR 3 , Y(CR 5 R 6 )C(O)NR 3 R 4 , Y(CR 5 R 6 )(CR 7 R 8 )C(O)R 3 , Y(CR 5 R 6 )(CR 7 R 8 )C(O)OR 3 , Y(CR 5 R 6 )(CR 7 R 8 )C(O)NR 3 R 4 , (CR 5 R 6 )NR 3 SO 2 R 3 or C(=NOR 3 )R 3 etc.; preferably, it is a benzene ring, C 1-6 alkoxy, Y(CR 5 R 6 )(CR 7 R 8 )OR 3; wherein, the heterocycloalkyl group includes a C cycloalkyl group containing 1-5 nitrogen, oxygen, or sulfur atoms; 3-8 The aryl group is selected from phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group is selected from pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl group, heterocycloalkyl group, aryl group, and heteroaryl group may be further substituted by 1-2 R 3 groups;

[0096] Y is O or CR 9 R 10 ;

[0097] R 3 is selected from H, C 1-6 alkyl, C 1-6 alkyl NHC(O)R 4 、C 1-6 alkyl C(O)NHR 4 、p(C 2 alkyloxy)C 2 alkyl NHC(O)R 4 、p(C 2 alkyloxy)C 2 alkyl C(O)NHR 4 etc.; preferably, it is H, C 1-6 alkyl C(O)NHR 4 ; where p is 0, 1, 2, 3, 4;

[0098] R 4 is H, C 1-6 alkyl, C 3-8 cycloalkyl, C 7-10 bridged cycloalkyl, heterocycloalkyl, aryl, heteroaryl, C 1-6 alkyl-cycloalkyl, C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-O-aryl, C 1-6 alkyl-heteroaryl, heteroaryl-aryl, etc.; preferably, it is C 1-6 alkyl-heterocycloalkyl, C 1-6 alkyl-aryl, C 1-6 alkyl-heteroaryl; wherein, the heterocycloalkyl group includes a C 3-8Cycloalkyl; the aryl group includes phenyl, anthryl, naphthyl, phenanthryl, etc.; the heteroaryl group includes pyridyl, furyl, pyrazolyl, pyranyl, pyridazinyl, pyrimidinyl, imidazolyl, thiazolyl, isothiazolyl, piperazinyl, quinolinyl, pyrazinyl, isoquinolinyl, thienyl, oxazolyl, isoxazolyl, indolyl, benzofuranyl, benzothienyl, purinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, pteridinyl, etc.; wherein, the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups may be further substituted by 1-2 R 3 groups;

[0099] R 5 、R 6 、R 7 、R 8 、R 9 and R 10 are independently selected from H, halogen, C 1-6 alkyl, OH, CO 2 R 3 、NR 3 R 4 、C 1-6 alkyloxy, etc.; preferably, it is H;

[0100] The compound as described in formula (I)-(VII) or its pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer, and the compound can specifically be selected from:

[0101] N-([1,1'-Biphenyl]-4-yl)thiophene-2-carboxamide;

[0102] N-([1,1'-Biphenyl]-3-yl)thiophene-2-carboxamide;

[0103] N-([1,1'-Biphenyl]-2-yl)thiophene-2-carboxamide;

[0104] 5-([1,1'-Biphenyl]-4-ylcarbamoyl)thiophene-2-carboxylic acid;

[0105] 5-([1,1'-Biphenyl]-3-ylcarbamoyl)thiophene-2-carboxylic acid;

[0106] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)furan-2-carboxamide;

[0107] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-1H-pyrrole-2-carboxamide;

[0108] 5-((5-Chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxylic acid;

[0109] N2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(3-hydroxypropyl)thiophene-2,5-dicarboxamide;

[0110] N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(4-hydroxybutyl)thiophene-2,5-dicarboxamide;

[0111] N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(6-hydroxyhexyl)thiophene-2,5-dicarboxamide;

[0112] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-nitrothiophene-2-carboxamide;

[0113] 5-amino-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0114] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(2-chloroacetamido)thiophene-2-carboxamide;

[0115] N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -cyclopropylthiophene-2,5-dicarboxamide;

[0116] N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(2-hydroxyethyl)thiophene-2,5-dicarboxamide;

[0117] N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(5-(5-((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)pentyl)thiophene-2,5-dicarboxamide;

[0118] N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N5-(2-(2-hydroxyethoxy)ethyl)thiophene-2,5-dicarboxamide;

[0119] 5-(2-aminoacetamido)-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0120] N-(2-chloro-5-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0121] N-(4-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0122] N-([1,1'-biphenyl]-3-yl)thiophene-3-carboxamide;

[0123] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(5-((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)hexanamido)thiophene-2-carboxamide;

[0124] N-(4-(2-methoxyethyl)phenyl)thiophene-2-carboxamide;

[0125] N-(3-(2-methoxyethyl)phenyl)thiophene-2-carboxamide;

[0126] N-(3-(1-methyl-1H-pyrazol-4-yl)phenyl)thiophene-2-carboxamide;

[0127] N-(3-(1H-pyrazol-4-yl)phenyl)thiophene-2-carboxamide;

[0128] N-(3-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0129] N-(2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0130] N-(3-(1-(2-hydroxyethyl)-1H-pyrazol-4-yl)phenyl)thiophene-2-carboxamide;

[0131] N-([[1,1'-biphenyl]-3-yl)-5-(6-(5-((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)hexanamido)thiophene-2-carboxamide;

[0132] N2-([1,1'-biphenyl]-3-yl)-N5-(5-(5-(((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)pentyl)thiophene-2,5-dicarboxamide;

[0133] N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(((2R,3R,4S,5R)-3,4,5,6-tetrahydroxy-1-oxohexan-2-yl)thiophene-2,5-dicarboxamide;

[0134] N-(1-(2-Hydroxyethyl)-1H-pyrazol-4-yl)thiophene-2-carboxamide;

[0135] N-(5-Methyl-1H-pyrazol-3-yl)thiophene-2-carboxamide;

[0136] N-(1-Methyl-1H-pyrazol-4-yl)thiophene-2-carboxamide;

[0137] N-([1,1'-Biphenyl]-3-yl)-1H-1,2,4-triazole-5-carboxamide;

[0138] N-([1,1'-Biphenyl]-3-yl)thiazole-2-carboxamide;

[0139] N-([1,1'-Biphenyl]-3-yl)thiazole-4-carboxamide;

[0140] N-([1,1'-Biphenyl]-3-yl)-1H-pyrazole-5-carboxamide;

[0141] N-([1,1'-Biphenyl]-3-yl)thiazole-5-carboxamide;

[0142] N-([1,1'-Biphenyl]-3-yl)-1-hydroxy-1H-1,2,3-triazole-4-carboxamide;

[0143] N-([1,1'-Biphenyl]-3-yl)isothiazole-5-carboxamide;

[0144] N-(Thiophen-2-yl)-[1,1'-biphenyl]-3-carboxamide;

[0145] N-([1,1'-Biphenyl]-3-yl)isothiazole-5-carboxamide;

[0146] 1-([1,1'-Biphenyl]-3-yl)-3-(thiophen-2-yl)urea;

[0147] N-([1,1'-Biphenyl]-3-ylmethyl)thiophene-2-carboxamide;

[0148] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)thiazole-5-carboxamide;

[0149] N-(4-Chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0150] N-(3-Chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0151] 4-Bromo-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0152] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-1-methyl-1H-pyrazole-3-carboxamide;

[0153] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(methylsulfonyl)thiophene-2-carboxamide;

[0154] 3-bromo-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0155] N-(5-chloro-2-(2-hydroxyethyl)phenyl)thiophene-2-carboxamide;

[0156] N-(5-chloro-2-(pentyloxy)phenyl)thiophene-2-carboxamide;

[0157] N-(5-chloro-2-ethoxyphenyl)thiophene-2-carboxamide;

[0158] 4-chloro-2-(thiophene-2-carboxamido)phenylacetic acid;

[0159] N-(2-(2-methoxyethoxy)-5-methylphenyl)thiophene-2-carboxamide;

[0160] N-(2-butoxy-5-chlorophenyl)thiophene-2-carboxamide;

[0161] N-(5-chloro-2-propoxyphenyl)thiophene-2-carboxamide;

[0162] N-(5-chloro-2-(3-methoxypropoxy)phenyl)thiophene-2-carboxamide;

[0163] N-(5-chloro-2-(2-ethoxyethoxy)phenyl)thiophene-2-carboxamide;

[0164] N-(5-chloro-2-(2-hydroxyethoxy)phenyl)thiophene-2-carboxamide;

[0165] N-(5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)thiophene-2-carboxamide;

[0166] N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiazole-2-carboxamide;

[0167] N-(5-chloro-2-(heptyloxy)phenyl)thiophene-2-carboxamide;

[0168] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-6-ethynylnicotinamide;

[0169] Methyl 4-chloro-2-(thiophene-2-carboxamido)benzoate;

[0170] N-(2-(2-Methoxyethyl)phenyl)thiophene-2-carboxamide;

[0171] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-(cyclopropanecarboxamido)thiophene-2-carboxamide;

[0172] N-(5-Chloro-2-(prop-2-yn-1-yloxy)phenyl)thiophene-2-carboxamide;

[0173] (5-((5-Chloro-2-(2-methoxyethoxy)phenylcarbamoyl)thiophene-2-carbonyl)glycine;

[0174] N-(5-Methoxy-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0175] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-methoxypyrazine-2-carboxamide;

[0176] N-(5-Bromo-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0177] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-4-phenylthiazole-2-carboxamide;

[0178] N-(5-Fluoro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0179] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-sulfonamide;

[0180] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-1-hydroxy-1H-benzo[d][1,2,3]triazole-6-carboxamide;

[0181] tert-Butyl (5-(5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxamido)pentyl)carbamate;

[0182] N 2 -(5-Aminopentyl)-N 5 -(5-Chloro-2-(2-methoxyethoxy)phenyl)thiophene-2,5-dicarboxamide;

[0183] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-methoxypicolinamide;

[0184] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(2-(2-(2-(5-(((3aR,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)ethoxy)ethoxy)acetamido)thiophene-2-carboxamide;

[0185] N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(2-(2-(2-(5-(((3aR,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamidoethoxy)ethoxy)ethylthiophene-2,5-dicarboxamide;

[0186] N-(5-chloro-2-(2-methoxyethoxy)phenyl)pyrimidine-4-carboxamide;

[0187] N-(5-chloro-2-(2-methoxyethoxy)phenyl)pyridazine-3-carboxamide;

[0188] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-2-phenylcyclopropane-1-carboxamide;

[0189] N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiazole-4-carboxamide;

[0190] N-(5-chloro-2-(2-methoxyethoxy)phenyl)isothiazole-5-carboxamide;

[0191] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-1H-1,2,4-triazole-5-carboxamide;

[0192] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-4-hydroxypyrrolidine-2-carboxamide;

[0193] 2-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene;

[0194] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-3-(thiophen-2-yl)propanamide;

[0195] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-4-(thiophen-2-yl)butanamide;

[0196] N-([[1,1'-biphenyl]-3-yl)-2-(thiophen-2-yl)acetamide;

[0197] N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5-(2-(2-(2-(2-(2-Phenylacetamido)ethoxy)ethoxy)ethyl)thiophene-2,5-dicarboxamide;

[0198] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(thiophene-2-carboxamido)hexanamido)thiophene-2-carboxamide;

[0199] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(2-phenylacetamido)hexanamido)thiophene-2-carboxamide;

[0200] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(2-(thiophen-2-yl)acetamido)hexanamido)thiophene-2-carboxamide;

[0201] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(3-(4-fluorophenyl)propanamido)hexanamido)thiophene-2-carboxamide;

[0202] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(4-(thiophen-2-yl)butanamido)hexanamido)thiophene-2-carboxamide;

[0203] 2-Chloro-N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)pyrimidine-5-carboxamide;

[0204] 3-Bromo-N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)furan-2-carboxamide;

[0205] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(4-chloro-2-methylbenzamido)hexanamido)thiophene-2-carboxamide;

[0206] N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(3-(thiophen-2-yl)propanamido)hexanamido)thiophene-2-carboxamide;

[0207] N 2 -(5-Chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)thiophene-2,5-dicarboxamide;

[0208] N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)benzofuran-2-carboxamide;

[0209] N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)thiazole-5-carboxamide;

[0210] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(2-(pyridin-2-yl)acetamido)hexanamido)thiophene-2-carboxamide;

[0211] N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)-1H-pyrazole-5-carboxamide;

[0212] 5-(6-(2-(1H-tetrazol-1-yl)acetamido)hexanamido)-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide;

[0213] N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(4-phenylbutyramido)hexanamido)thiophene-2-carboxamide.

[0214] The present invention also provides a pharmaceutical composition, which comprises the compound or its pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer described above in the present invention, and a pharmaceutically acceptable excipient.

[0215] In certain embodiments, the pharmaceutical composition further comprises an additional compound that can effectively prevent, treat, or improve arthritis or joint injury and / or symptoms associated with arthritis or joint injury in mammals.

[0216] In certain embodiments, the additional compound is selected from one or more of NSAIDs, analgesics, angiopoietin-like 3 protein (ANGPTL3) or its chondrogenic variant, oral salmon calcitonin, SD-6010 (iNOS inhibitor), vitamin D3 (cholecalciferol), apoptosis / caspase inhibitor (enlimomab), collagen hydrolysate, FGF18, BMP7, avocado soy unsaponifiables (ASU), hyaluronic acid; preferably, FGF18, BMP7, hyaluronic acid.

[0217] The present invention also provides the use of the aforementioned compound or its pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer, or pharmaceutical composition in the preparation of a pharmaceutical composition for preventing and / or treating and / or ameliorating arthritis or joint injury and / or symptoms associated with arthritis or joint injury in mammals.

[0218] The arthritis is one or more of osteoarthritis, traumatic arthritis, autoimmune arthritis, etc.

[0219] The compound or its pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer, pharmaceutical composition can be administered to a desired subject in a matrix and a biocompatible scaffold; the administered composition also contains hyaluronic acid.

[0220] The present invention also provides a method for preventing, treating, or ameliorating arthritis or joint injury in mammals, the method comprising administering to the mammal a therapeutically effective amount of the compound or composition of the present invention, wherein the mammal has joint damage or arthritis or is at risk thereof.

[0221] The present invention also provides a method for preventing, treating, or ameliorating arthritis or joint injury in a human patient, the method comprising: administering to the joint of the patient a composition comprising an effective amount of the compound or composition of the present invention, thereby treating, ameliorating, or preventing arthritis or joint injury in the patient.

[0222] The patient has arthritis or joint injury; or does not have arthritis or joint injury, but is at risk thereof.

[0223] Wherein, the arthritis is osteoarthritis, traumatic arthritis, autoimmune arthritis; the administered composition also contains hyaluronic acid.

[0224] The present invention also provides a method for inducing mesenchymal stem cells (MSCs) to differentiate into chondrocytes using the aforementioned compound. The method includes contacting mesenchymal stem cells with a sufficient amount of the compound of the present invention or its pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer, or pharmaceutical composition, thereby inducing the stem cells to differentiate into chondrocytes.

[0225] The method for inducing mesenchymal stem cells (MSCs) to differentiate into chondrocytes can be carried out in vitro or in a mammal, and the stem cells are present in the mammal.

[0226] In some embodiments, the mammal is a domesticated animal or livestock. In certain embodiments, the mammal is a human, mouse, rabbit, dog, cat or horse.

[0227] The contact described in the above method is carried out in a matrix or a biocompatible scaffold; the contact can also be carried out by binding the compound to one or more additional chondrogenic factors; the contact can also be carried out by binding the compound to an active agent selected from angiopoietin-like 3 protein (ANGPTL3), oral salmon calcitonin, SD-6010 (iNOS inhibitor), vitamin D3 (cholecalciferol), collagen hydrolysate, FGF18, BMP7, romosozumab, avocado soybean unsaponifiables (ASU), steroids and steroidal anti-inflammatory drugs (NSAIDs), hyaluronic acid, and the like, one or several of them.

[0228] Any suitable amount of the compound of the present invention or its pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer, or pharmaceutical composition can induce the differentiation of MSCs into chondrocytes. Depending on the specific application and efficacy of the active ingredient, the amount of the compound of the present invention or its pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer, or pharmaceutical composition used ranges from 0.1 mg to 10,000 mg, such as an amount of 1.0 mg to 1000 mg, such as an amount of 10 mg to 5000 mg, etc. In some embodiments, in intra-articular injection of the knee, the compound of the present invention can be present at a concentration of 0.1 μM to about 100 μM.

[0229] The compound provided by the present invention or its pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer, or pharmaceutical composition can be used to treat, improve or prevent any type of articular cartilage damage (such as joint injury or trauma), including, for example, damage resulting from a traumatic event or tendon or ligament tear. In some embodiments, administering the compound of the present invention or its pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer, or pharmaceutical composition can prevent or improve arthritis or joint damage, for example, where there is a genetic or family history of arthritis or joint damage or joint injury before or during joint surgery; it can also be used to treat joint damage; the joint damage is traumatic joint damage; it can also be damage caused by age or inactivity; it can also be damage caused by an autoimmune disease. In some embodiments of the present invention, the compounds, compositions and methods of the present invention can be used to treat, improve or prevent osteoarthritis (Example 3-1). In some embodiments, the compounds, compositions and methods are used to improve or prevent arthritis in subjects at risk of having or acquiring arthritis (Examples 3-2, 3). In some embodiments, the compounds, compositions and methods are used to improve or prevent joint injury in subjects at risk of having or acquiring joint injury (Examples 3-2, 3-3).

[0230] The compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions provided by the present invention can be used in a method for stimulating chondrocyte proliferation and cartilage production in damaged cartilage tissue caused by, for example, traumatic injury or chondropathy. Specifically, the compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions provided by the present invention can be used for treating cartilage damage in joints, such as cartilage damage on joint articular surfaces, such as in the spine, acromion, wrist, finger, hip, knee, ankle joints and foot joints; examples of diseases or disorders that can benefit from treatment include osteoarthritis, rheumatoid arthritis, other autoimmune diseases or osteochondritis dissecans. In addition, cartilage damage or destruction can also occur as a result of some genetic or metabolic disorders, cartilage deformities are usually observed in human dwarfism, and / or treatment of cartilage damage or destruction is usually the result of reconstructive surgery; therefore, the aforementioned compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions provided by the present invention, whether used alone or in combination with other methods, are useful therapies for patients.

[0231] The compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof provided by the present invention, or pharmaceutical compositions thereof, can be administered by direct injection into the synovial fluid of the joint, systemic administration (oral or intravenous), or direct injection into the cartilage defect, and can be administered alone or in combination with a suitable carrier for extended protein release; in some embodiments, the compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions can be administered in a biocompatible material or scaffold. The compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions of the present invention can also be used in combination with surgical procedures at the damaged joint. The implementation of the present invention can be carried out before, during or in combination with, and / or after the surgical procedure. For example, the compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions of the present invention can be used to expand the chondrocyte population in culture for autologous or allogeneic chondrocyte implantation (ACI); chondrocytes can be treated while administering the compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions of the present invention. In these methods, for example, chondrocytes can be collected from an area of the damaged joint with a small amount of undamaged load through arthroscopic surgery optionally in the presence of the compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions of the present invention and / or additional growth factors, and incubated in vitro to increase the cell number, and then transplanted. Then, optionally, the expanded culture is mixed with the compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions of the present invention and placed into the joint space or directly into the defect. In some embodiments, the expanded culture can be placed into the joint space suspended in a matrix or membrane together with the compounds of the present invention. In additional embodiments, the compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions of the present invention can also be used in combination with one or more perichondrium or perichondrium grafts containing chondrogenic cells to help keep the implanted chondrocytes or chondrocyte precursor cells in place. In some embodiments, the use of the compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions of the present invention in combination with other methods can also repair cartilage damage, and the other methods include but are not limited to joint lavage, bone marrow stimulation, abrasion arthroplasty, subchondral drilling or microfracture of the proximal subchondral bone, etc.After administering the compound of the present invention or a pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer thereof, or a pharmaceutical composition, and after cartilage growth, surgical treatment can be beneficial to appropriately contour the newly formed cartilage surface.

[0232] The compound of the present invention or a pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer thereof, or a pharmaceutical composition can promote collagen expression in human epidermal fibroblasts; the present invention thus provides a method for increasing collagen production in fibroblasts, which is carried out by contacting fibroblasts with the compound of the present invention or a pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer thereof, or a pharmaceutical composition, thereby increasing the production of collagen in fibroblasts. The contacting can be carried out in vivo by directly injecting the compound into the area to be treated, or can be carried out in vitro on a population of fibroblasts. Brief Description of the Drawings

[0233] Figure 1 It is a schematic diagram of the cartilage injury repair effect of PA002 of the present invention on a C57 mouse acute injury surgical model. Among them, Figure 1 A is the safranin-fast green staining result of paraffin sections of the knee joints of mice; Figure 1 B is the statistical result of the joint score of the lateral tibial plateau with cartilage injury; Figure 1 C is the result of evaluating the pain relief of PA002 on OA-induced pain in mice by weight-bearing test; Figure 1 D is the result of evaluating the pain relief of PA002 on OA-induced pain in mice by hot plate experiment.

[0234] Figure 2 It is a schematic diagram of the cartilage repair effect of PA002 of the present invention on a C57 mouse cartilage defect model. Among them, Figure 2 A is the repair situation of cartilage defects at the joints of mice under microscopic imaging (the circled area is the cartilage defect site); Figure 2 B is the safranin-fast green staining result of mouse joint sections.

[0235] Figure 3 It is a schematic diagram of the cartilage repair effect of PA002 of the present invention on an SD rat cartilage defect model. Among them, Figure 3 A is the cartilage defect repair effect in vivo induced by PA002 in vitro on UC-MSCs and the comparison with the control group; Figure 3 B is the effect of PA002-induced hydrogel-encapsulated UC-MSCs on repairing cartilage defects in vivo and the comparison with the control group. Detailed Description of the Embodiments

[0236] The invention will be further described in detail below in conjunction with specific embodiments and the accompanying drawings. The processes, conditions, experimental methods, etc. for implementing the present invention are common knowledge and well-known common sense in the art except for the specifically mentioned content below, and the present invention has no particularly restricted content.

[0237] The present invention provides a class of compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions, which can stimulate chondrocyte differentiation in mesenchymal stem cells. The present invention also provides a method for inducing the differentiation of mesenchymal stem cells into chondrocytes. Further, the present invention provides for the treatment, prevention or improvement of arthritis or joint injuries and / or symptoms associated with arthritis or joint injuries by administering the compounds of the present invention or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions into joints, vertebrae, intervertebral discs or systemically.

[0238] In the following description, certain specific details are set forth in order to provide a thorough understanding of the embodiments. However, those skilled in the art will understand that the present invention may be practiced without these details. In other instances, well-known structures have not been shown or described in detail to avoid an unnecessary obscuring description of the embodiments. Unless the context requires otherwise, throughout the specification and the claims that follow, the word "comprise" and its variants, such as "comprises" and "comprising", shall be construed in an open, inclusive sense, i.e., as "including but not limited to". Additionally, the headings provided in the present invention are for convenience only and do not interpret the scope or meaning of the claimed invention.

[0239] Unless otherwise specified, the following terms as used herein have the following meanings:

[0240] "Amino" refers to the -NH 2 group.

[0241] "Cyano" or "nitrile" refers to the -CN group.

[0242] "Hydroxy (Hydroxy or hydroxyl)" refers to the -OH group.

[0243] "Nitro" refers to the -NO 2 group.

[0244] "Oxo" refers to the =O substituent.

[0245] "Oxime" refers to the =N-OH substituent.

[0246] "Thio" refers to the =S substituent.

[0247] "Alkyl" means a straight-chain or branched-chain hydrocarbon chain group that is fully saturated or contains unsaturated bonds, having 1 to 30 carbon atoms, and is connected to the rest of the molecule by a single bond. Alkyl groups containing any number of carbon atoms from 1 to 30 are encompassed. An alkyl group containing up to 30 carbon atoms is referred to as a C 1-30 alkyl group. Similarly, for example, an alkyl group containing up to 12 carbon atoms is a C 1-12 alkyl group. Alkyl groups containing other numbers of carbon atoms (and other moieties defined in the present invention) are represented in a similar manner.

[0248] "Aryl" means a group derived from a hydrocarbon ring system containing hydrogen, 6 to 30 carbon atoms, and at least one aromatic ring. The aryl group can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, and can include fused or bridged ring systems. Unless otherwise specifically stated in the specification, the term "aryl" or the prefix "ar-" (as in "aralkyl") is intended to include optionally substituted aryl groups.

[0249] "Cycloalkyl" or "carbocyclic" means a stable, non-aromatic monocyclic or polycyclic carbocyclic ring, which can include fused or bridged ring systems, and which is saturated or unsaturated. Representative cycloalkyl or carbocyclic rings include, but are not limited to, cycloalkyl rings having 3 to 15 carbon atoms, 3 to 10 carbon atoms, 3 to 8 carbon atoms, 3 to 6 carbon atoms, 3 to 5 carbon atoms, or 3 to 4 carbon atoms. Unless otherwise specifically stated in the specification, the cycloalkyl or carbocyclic ring can be optionally substituted.

[0250] "Fused" means any ring structure described herein that is fused to an existing ring structure. When the fused ring is a heterocyclic ring or a heteroaryl ring, any carbon atom on the existing ring structure that becomes part of the fused heterocyclic ring or fused heteroaryl ring can be replaced by a nitrogen atom.

[0251] "Halogen (halo)" or "halogen" means bromine, chlorine, fluorine, or iodine.

[0252] "Haloalkyl" means an alkyl group as defined above that is substituted with one or more halogen groups as defined above.

[0253] Similarly, "haloalkoxy" means a group of the formula -OR a wherein R a is a haloalkyl group as defined above. Unless otherwise specifically stated in the specification, the haloalkoxy group can be optionally substituted as described below.

[0254] "Heterocycloalkyl", "heterocyclic group", "heterocyclic ring", or "heterocycle" refers to a stable 3- to 24-membered non-aromatic ring group containing 2 to 23 carbon atoms and 1 to 8 heteroatoms selected from nitrogen, oxygen, phosphorus, and sulfur. Unless otherwise specifically stated in the specification, the heterocyclic group can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, and can include fused or bridged ring systems; and the nitrogen, carbon, or sulfur atoms in the heterocyclic group can be optionally oxidized; the nitrogen atoms can be optionally quaternized; and the heterocyclic group can be partially or fully saturated. Unless otherwise specifically stated in the specification, the heterocyclic group can be optionally substituted. Illustrative examples of heterocycloalkyl are also referred to as non-aromatic heterocycles. The term heterocycloalkyl also includes all cyclic forms of carbohydrates, including but not limited to monosaccharides, disaccharides, and oligosaccharides. Unless otherwise stated, heterocycloalkyl has 2 to 10 carbons in the ring. It should be understood that when referring to the number of carbon atoms in a heterocycloalkyl, the number of carbon atoms in the heterocycloalkyl is different from the total number of atoms (including heteroatoms) that make up the heterocycloalkyl (i.e., the backbone atoms of the heterocycloalkyl ring). Unless otherwise specifically stated in the specification, heterocycloalkyl can be optionally substituted.

[0255] "Heteroaryl" refers to a 5- to 14-membered ring system group containing a hydrogen atom, 1 to 13 carbon atoms, 1 to 6 heteroatoms selected from nitrogen, oxygen, phosphorus, and sulfur, and at least one aromatic ring. For the purposes of the present invention, the heteroaryl can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, and can include fused or bridged ring systems; and the nitrogen, carbon, or sulfur atoms in the heteroaryl can be optionally oxidized; the nitrogen atoms can be optionally quaternized. Unless otherwise specifically stated in the specification, the heteroaryl can be optionally substituted.

[0256] The term "optional" or "optionally" means that the subsequent described event or situation may or may not occur, and the description includes the case where the event or situation occurs and the case where the event or situation does not occur.

[0257] "Effective amount" or "therapeutically effective amount" refers to the amount of a compound that is administered to a mammalian subject as a single dose or as part of a series of doses and effectively produces the desired therapeutic effect.

[0258] "Treatment" of an individual (e.g., a mammal, such as a human) or a cell is any type of intervention used in an attempt to alter the natural course of the individual or cell. In some embodiments, treatment includes administering a pharmaceutical composition after the onset of a pathological event or contact with a pathogen, and includes stabilization of the condition (e.g., the condition does not worsen or the condition is alleviated). In other embodiments, treatment also includes prophylactic treatment (e.g., administering the compositions described herein when an individual is suspected of having a bacterial infection).

[0259] "Tautomer" refers to the transfer of a proton from one atom of a molecule to another atom of the same molecule. The compounds provided herein can exist as tautomers. Tautomers are compounds that can be interconverted by the migration of a hydrogen atom (accompanied by the conversion of a single bond and an adjacent double bond). In the bonding arrangements where tautomerism can occur, there will be a chemical equilibrium of tautomers. All tautomeric forms of the compounds disclosed herein are considered. The exact ratio of tautomers depends on several factors, including temperature, solvent, and pH.

[0260] A "metabolite" of a compound disclosed in the present invention is a derivative of the compound formed when the compound is metabolized. The term "active metabolite" refers to a bioactive derivative of the compound formed when the compound is metabolized. As used herein, the term "metabolism" refers to the sum of the processes by which a particular substance is altered by an organism. Thus, enzymes can produce specific structural changes in a compound. The metabolites of the compounds disclosed herein can be identified by administering the compound to a host and analyzing tissue samples from the host, or by incubating the compound with hepatocytes in vitro and analyzing the resulting compounds. Both of these methods are well known in the art. In some embodiments, the metabolites of the compound are formed by an oxidation process and correspond to the corresponding hydroxy-containing compounds. In some embodiments, the compound is metabolized into a pharmacologically active metabolite.

[0261] The present invention provides compounds for inducing the differentiation of mesenchymal stem cells into chondrocytes and for preventing, treating, and improving arthritis or joint injuries and / or symptoms associated with arthritis or joint injuries in mammals, as well as methods for preparing such compounds. The present invention also provides such compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers, or isomers thereof, and pharmaceutical compositions comprising at least one such compound or pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof and a pharmaceutically acceptable excipient.

[0262] The compounds provided by the present invention can be synthesized using standard synthetic reactions known to those skilled in the art or using methods known in the art. The reactions can be employed in a linear sequence to provide the compounds, or the reactions can be used to synthesize fragments, which are then joined by methods known in the art.

[0263] If desired, conventional techniques including, but not limited to, filtration, distillation, crystallization, and chromatography can be used to isolate and purify the reaction products. Such materials can be characterized using conventional means including physical constants and spectroscopic data.

[0264] The low-temperature reaction apparatus used in the present invention is an EYELA (PSL-1810) magnetic stirring low-temperature constant temperature water bath; an EYELA (N-1100) rotary evaporator; the purity results of the compounds obtained in the experiments are all from an Agilent 1200 series LCsystem high performance liquid chromatograph analyzer (chromatographic conditions: Zorbax XDB-C18 (4.6×150 mm, 5 μm), column temperature 40 °C, mobile phase MeOH / H 2 O, running flow rate 1.5 mL / min, UV detection wavelength 254 nm, injection volume 10 μL); the nuclear magnetic resonance spectrometer is a Bruker 300 type or 500 type (internal standard: TMS, solvent used is CDCl 3 or DMSO-d 6 ); the purification of reaction intermediates and end products is all carried out using a chromatography column (the silica gel used is 200-300 mesh), and the silica gel used is purchased from Qingdao Ocean Chemical Factory. All solvents are redistilled before use, and the anhydrous solvents used are all obtained by drying according to standard methods; unless otherwise specified, all reactions are carried out under nitrogen protection and the reaction progress is tracked by TLC, and the post-treatment is all washed with saturated brine and dried over anhydrous sodium sulfate.

[0265] The mammalian part does not have arthritis or joint injury, but has an increased risk of developing arthritis or joint injury.

[0266] The compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions provided by the present invention can be used for preventing, treating, improving any type of arthritis or joint injury or various cartilage disorders; in some embodiments, the compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions of the present invention are administered to prevent arthritis or joint injury, for example, in the presence of a genetic or family history of arthritis or joint injury or previous joint surgery, or in other situations where the risk of arthritis or joint injury is increased. Exemplary conditions or disorders that the compounds or pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers thereof, or pharmaceutical compositions provided by the present invention can treat or prevent include but are not limited to systemic rheumatoid arthritis, juvenile chronic arthritis, osteoarthritis, intervertebral disc degenerative disease, spondyloarthropathy and systemic sclerosis (scleroderma). The arthritis can be osteoarthritis, traumatic arthritis, intervertebral disc degenerative disease, Dupuytren's disease or tendon disease.

[0267] The present invention also provides a method for stimulating chondrocyte proliferation and cartilage production in cartilage tissue damaged due to traumatic injury or chondropathy using the aforementioned compound or its pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer, or pharmaceutical composition. Traumatic injuries can include, but are not limited to, blunt trauma to joints or ligament injuries, such as anterior cruciate ligament, medial collateral ligament tears or meniscus tears; examples of tissues that exhibit joint articulation surfaces and are thus particularly sensitive to treatment include, but are not limited to, the spine, shoulder, elbow, wrist, finger joints, hip, knee, ankle, foot joints, etc. Examples of diseases that can benefit from the treatment include osteoarthritis, rheumatoid arthritis, other autoimmune diseases, osteochondritis dissecans, etc. Additionally, cartilage deformities are common in human dwarf forms, and thus the present invention can be used to assist in treating patients with cartilage deformities.

[0268] As used herein, "mammal" refers to any mammal classified as a mammal, including humans, domesticated animals and livestock, companion animals or farm animals, and animals kept in zoos or used in competitive events; "companion animal" refers to dogs, cats, rodents and rabbits. Specifically, it can be a human, a cow (e.g., a female cow), a horse, a dog, a sheep, a pig, a rabbit, a goat, a cat, etc. The choice of animal is determined according to the actual situation.

[0269] The present invention also provides a method for inducing the differentiation of mesenchymal stem cells (MSCs) into chondrocytes. The method comprises contacting the mesenchymal stem cells with a sufficient amount of the compound of the present invention or its pharmaceutically acceptable salt, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer or isomer, or pharmaceutical composition, thereby inducing the differentiation of the stem cells into chondrocytes.

[0270] MSCs are pluripotent stem cells that can differentiate into several different types of cells, including but not limited to osteoblasts, chondrocytes and adipocytes. The differentiation process is the formation of a specialized cell type from a less specialized cell type, such as the formation of chondrocytes from MSCs. The method for inducing the differentiation of mesenchymal stem cells (MSCs) into chondrocytes can be carried out in vitro or in vivo in a mammal, and the stem cells are present in the mammal. In certain embodiments, the mammal is one or more of a human, a dog, a cat or a horse, etc.

[0271] The differentiation of MSCs into chondrocytes can be achieved using any appropriate amount of the compounds of the present invention or their pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers, or pharmaceutical compositions. Depending on the specific application and potency of the active ingredient, the amount of the compounds of the present invention or their pharmaceutically acceptable salts, polymorphs, prodrugs, esters, metabolites, N-oxides, stereoisomers or isomers, or pharmaceutical compositions used ranges from 0.1 mg to 10,000 mg, such as 1.0 mg to 1000 mg or for example 10 mg to 500 mg, etc. In some embodiments, the compounds of the present invention may also be present in an intra-articular injection for the knee at a concentration of 0.1 μM - 100 μM.

[0272] Example 1: Preparation of each compound

[0273] Example 1-1: Preparation of N-([1,1'-biphenyl]-4-yl)thiophene-2-carboxamide (PA001)

[0274] 4-Aminobiphenyl (169 mg, 1 mmol), 2-thiophenecarboxylic acid (141 mg, 1.1 mmol), N,N-diisopropylethylamine (388 mg, 3 mmol) and HATU (418 mg, 1.1 mmol) were dissolved in 3 mL of N,N-dimethylformamide and stirred at room temperature for 6 h. After TLC detection showed that the reaction was complete, the reaction solution was extracted with ethyl acetate. The organic phase was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After evaporating the organic phase, the product PA001 (220 mg, 79%) was obtained by column chromatography separation and purification.

[0275] 1 1H NMR (500 MHz, Chloroform-d) δ 7.72 (s, 1H), 7.70 (d, J = 8.6 Hz, 2H), 7.65 (dd, J = 3.8, 1.2 Hz, 1H), 7.63–7.58 (m, 4H), 7.56 (dd, J = 5.0, 1.2 Hz, 1H), 7.47–7.41 (m, 2H), 7.37–7.31 (m, 1H), 7.15 (dd, J = 5.0, 3.7 Hz, 1H).

[0276] Example 1-2: Preparation of N-([1,1'-biphenyl]-3-yl)thiophene-2-carboxamide (PA002)

[0277] Using a method similar to the preparation of compound PA001, 4-aminobiphenyl was replaced with 3-aminobiphenyl, and finally compound PA002 was obtained with a yield of 75%.

[0278] 11H NMR (500 MHz, Chloroform-d) δ 7.90–7.86 (m, 1H), 7.75 (s, 1H), 7.67–7.64 (m, 1H), 7.61 (ddd, J=11.9, 8.6, 1.1 Hz, 3H), 7.56 (dd, J=5.0, 1.0 Hz, 1H), 7.47–7.41 (m, 3H), 7.42–7.32 (m, 2H), 7.15 (dd, J=4.9, 3.8 Hz, 1H).

[0279] Example 1-3, Preparation of N-([1,1'-Biphenyl]-2-yl)thiophene-2-carboxamide (PA003)

[0280] Using a method similar to the preparation of compound PA001, 4-aminobiphenyl was replaced with 2-aminobiphenyl, and finally compound PA003 was obtained with a yield of 77%.

[0281] 1 1H NMR (500 MHz, Chloroform-d) δ 8.50 (dd, J=8.3, 1.2 Hz, 1H), 7.89–7.85 (m, 1H), 7.57–7.50 (m, 2H), 7.50–7.35 (m, 5H), 7.30 (dd, J=7.6, 1.7 Hz, 1H), 7.24–7.18 (m, 1H), 7.16 (dd, J=3.8, 1.2 Hz, 1H), 7.02 (dd, J=5.0, 3.7 Hz, 1H).

[0282] Example 1-4, Preparation of 5-([1,1'-Biphenyl]-4-ylcarbamoyl)thiophene-2-carboxylic acid (PA004)

[0283] 4-Aminobiphenyl (169 mg, 1 mmol), methyl 5-carboxy-2-thiophenecarboxylate (205 mg, 1.1 mmol), N,N-diisopropylethylamine (388 mg, 3 mmol) and HATU (418 mg, 1.1 mmol) were dissolved in 3 mL of N,N-dimethylformamide and stirred at room temperature for 6 h. After TLC detection showed that the reaction was complete, the reaction solution was extracted with ethyl acetate. The organic phase was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After evaporating the organic phase, the compound methyl 5-([1,1'-biphenyl]-4-ylcarbamoyl)thiophene-2-carboxylate (270 mg, yield 80%) was obtained by column chromatography separation and purification.

[0284] Compound methyl 5-([1,1'-biphenyl]-4-ylcarbamoyl)thiophene-2-carboxylate (270 mg, 0.8 mmol), LiOH·H 2O (336 mg, 8 mmol) was dissolved in methanol / water (8 mL / 2 mL), and the reaction was carried out at 80 °C for 2 h. After the reaction was completed by TLC detection, the organic solvent was removed by rotary evaporation. The aqueous phase was adjusted to acidity with dilute hydrochloric acid, and the filter cake was obtained by suction filtration and dried in a vacuum drying oven to obtain the final product PA004 (212 mg, yield 82%).

[0285] 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.24 (s, 1H), 7.84 (d, J = 8.4 Hz, 3H), 7.67 (dd, J = 7.9, 4.0 Hz, 4H), 7.48–7.42 (m, 2H), 7.37–7.30 (m, 1H), 7.27 (d, J = 3.7 Hz, 1H).

[0286] Preparation of Example 1-5, 5-([1,1'-biphenyl]-3-ylcarbamoyl)thiophene-2-carboxylic acid (PA005)

[0287] Using a method similar to the preparation of compound PA004, 4-aminobiphenyl was replaced with 3-aminobiphenyl, and finally compound PA005 was obtained with a yield of 85%.

[0288] 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.23 (s, 1H), 7.90–7.86 (m, 1H), 7.73 (s, 1H), 7.62 (ddd, J = 11.9, 8.6, 1.1 Hz, 3H), 7.52 (dd, J = 5.0, 1.0 Hz, 1H), 7.47–7.40 (m, 3H), 7.43–7.33 (m, 2H), 7.16 (dd, J = 4.9, 3.8 Hz, 1H).

[0289] Preparation of Example 1-6, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-methoxypicolinamide (PA006)

[0290] 4-Chloro-2-nitrophenol (1.74 g, 10 mmol), 2-bromoethyl methyl ether (4.17 g, 30 mmol), and potassium carbonate (4.14 g, 30 mmol) were dissolved in N,N-dimethylformamide (30 mL), and the reaction was carried out at 80 °C for 2 h. After the reaction was completed by TLC detection, the reaction solution was extracted with ethyl acetate. The organic phase was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After the organic phase was evaporated to dryness, the compound 4-chloro-1-(2-methoxyethoxy)-2-nitrobenzene (2.2 g, yield 95%) was obtained by column chromatography separation and purification

[0291] Dissolve 4-chloro-1-(2-methoxyethoxy)-2-nitrobenzene (2.2 g, 9.5 mmol) in methanol (30 mL), add palladium on carbon, introduce hydrogen gas, and react at room temperature for 2 hours. After detecting the completion of the reaction by TLC, filter it through diatomaceous earth by suction filtration, and rotary evaporate the filtrate to obtain compound 5-chloro-2-(2-methoxyethoxy)aniline (1.8 g, yield 94%).

[0292] Dissolve 5-chloro-2-(2-methoxyethoxy)aniline (202 mg, 1 mmol), 5-methoxypicolinic acid (168 mg, 1.1 mmol), N,N-diisopropylethylamine (388 mg, 3 mmol) and HATU (418 mg, 1.1 mmol) in 3 mL of N,N-dimethylformamide, and stir and react at room temperature for 6 hours. After detecting the completion of the reaction by TLC, extract the reaction solution with ethyl acetate. Wash the organic phase twice with water, once with saturated brine, and dry it over anhydrous sodium sulfate. After evaporating the organic phase to dryness, separate and purify the product PA006 by column chromatography, with a yield of 70%.

[0293] 1 H NMR (500 MHz, Chloroform-d) δ10.51 (s, 1H), 8.66 (d, J = 2.5 Hz, 1H), 8.32–8.18 (m, 2H), 7.33 (dd, J = 8.7, 2.9 Hz, 1H), 7.01 (dd, J = 8.7, 2.6 Hz, 1H), 6.86 (d, J = 8.7 Hz, 1H), 4.30–4.17 (m, 2H), 3.94 (s, 3H), 3.89–3.82 (m, 2H), 3.52 (s, 3H).

[0294] Preparation of Examples 1-7, N-(5-chloro-2-(2-methoxyethoxy)phenyl)pyrimidine-4-carboxamide (PA007)

[0295] Using a method similar to the preparation of compound PA006, replace 5-methoxypicolinic acid with pyrimidine-4-carboxylic acid, and finally obtain compound PA007 with a yield of 73%.

[0296] 1 H NMR (500 MHz, Chloroform-d) δ10.58 (s, 1H), 9.31 (d, J = 1.4 Hz, 1H), 9.04 (d, J = 5.0 Hz, 1H), 8.63 (d, J = 2.5 Hz, 1H), 8.21 (dd, J = 5.0, 1.4 Hz, 1H), 7.07 (dd, J = 8.7, 2.6 Hz, 1H), 6.89 (d, J = 8.7 Hz, 1H), 4.37–4.16 (m, 2H), 3.98–3.72 (m, 2H), 3.52 (s, 3H).

[0297] Example 1 - 8, Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)pyridazine-3-carboxamide (PA008)

[0298] Using a method similar to the preparation of compound PA006, substituting 5-methoxypicolinic acid with pyridazine-3-carboxylic acid, the compound PA008 was finally obtained with a yield of 70%.

[0299] 1 H NMR(500MHz,Chloroform-d)δ9.67(dd,J=2.4,1.3Hz,1H),9.45(dd,J=5.2,1.2Hz,1H),9.15(s,1H),8.56(d,J=2.5Hz,1H),7.97(dd,J=5.3,2.4Hz,1H),7.11(dd,J=8.7,2.5Hz,1H),6.98(d,J=8.6Hz,1H),4.27–4.18(m,2H),3.77–3.68(m,2H),3.40(s,3H).

[0300] Example 1 - 9, Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-2-phenylcyclopropane-1-carboxamide (PA009)

[0301] Using a method similar to the preparation of compound PA006, substituting 5-methoxypicolinic acid with 2-phenylcyclopropane-1-carboxylic acid, the compound PA009 was finally obtained with a yield of 78%.

[0302] 1 H NMR(500MHz,Chloroform-d)δ8.51(d,J=2.5Hz,1H),8.44(s,1H),7.30(t,J=7.6Hz,2H),7.24–7.19(m,1H),7.17–7.09(m,2H),6.96(dd,J=8.6,2.6Hz,1H),6.86(d,J=8.6Hz,1H),4.19–4.05(m,2H),3.72–3.63(m,2H),3.34(s,3H),2.60(ddd,J=9.0,6.5,4.0Hz,1H),1.81(ddd,J=8.1,5.2,4.1Hz,1H),1.73(dt,J=9.5,4.8Hz,1H),1.38(ddd,J=8.2,6.5,4.5Hz,1H).

[0303] Example 1 - 10, Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiazole-4-carboxamide (PA010)

[0304] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with thiazole-4-carboxylic acid, and finally compound PA010 was obtained with a yield of 69%.

[0305] 1 H NMR (500 MHz, Chloroform-d) δ 9.97 (s, 1H), 8.80 (d, J = 2.1 Hz, 1H), 8.62 (d, J = 2.6 Hz, 1H), 8.27 (d, J = 2.1 Hz, 1H), 7.03 (dd, J = 8.7, 2.6 Hz, 1H), 6.87 (d, J = 8.7 Hz, 1H), 4.28–4.17 (m, 2H), 3.86–3.78 (m, 2H), 3.49 (s, 3H).

[0306] Preparation of Example 1-11, N-(5-chloro-2-(2-methoxyethoxy)phenyl)isothiazole-5-carboxamide (PA011)

[0307] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with isothiazole-5-carboxylic acid, and finally compound PA011 was obtained with a yield of 72%.

[0308] 1 H NMR (500 MHz, Chloroform-d) δ 8.82 (s, 1H), 8.56 (d, J = 1.8 Hz, 1H), 8.51 (d, J = 2.6 Hz, 1H), 7.65 (d, J = 1.7 Hz, 1H), 7.07 (dd, J = 8.7, 2.5 Hz, 1H), 6.93 (d, J = 8.7 Hz, 1H), 4.32–4.12 (m, 2H), 3.78–3.66 (m, 2H), 3.41 (s, 3H).

[0309] Preparation of Example 1-12, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-1H-1,2,4-triazole-5-carboxamide (PA012)

[0310] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 1H-1,2,4-triazole-5-carboxylic acid, and finally compound PA012 was obtained with a yield of 74%.

[0311] 11H NMR (500 MHz, Chloroform-d) δ 9.73 (s, 1H), 8.54 (d, J = 2.5 Hz, 1H), 8.26 (s, 1H), 7.08 (dd, J = 8.7, 2.5 Hz, 1H), 6.91 (d, J = 8.7 Hz, 1H), 4.29–4.21 (m, 2H), 3.86–3.78 (m, 2H), 3.49 (s, 3H).

[0312] Example 1-13. Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-4-hydroxypyrrolidine-2-carboxamide (PA013)

[0313] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 4-hydroxypyrrolidine-2-carboxylic acid to finally obtain compound PA013 with a yield of 71%.

[0314] 1 1H NMR (500 MHz, Chloroform-d) δ 9.73 (s, 1H), 8.54 (d, J = 2.5 Hz, 1H), 8.26 (s, 1H), 7.08 (dd, J = 8.7, 2.5 Hz, 1H), 6.91 (d, J = 8.7 Hz, 1H), 4.29–4.21 (m, 2H), 3.86–3.78 (m, 2H), 3.49 (s, 3H).

[0315] Example 1-14. Preparation of 2-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene (PA014)

[0316] Dissolve 2-bromo-4-chlorophenol (207 mg, 1 mmol), 2-bromoethyl methyl ether (417 mg, 3 mmol), and potassium carbonate (414 mg, 3 mmol) in N,N-dimethylformamide (5 mL), and react at 80 °C for 2 h. After detecting the completion of the reaction by TLC, extract the reaction solution with ethyl acetate. Wash the organic phase twice with water, once with saturated brine, and dry over anhydrous sodium sulfate. After evaporating the organic phase, separate and purify by column chromatography to obtain compound 2-bromo-4-chloro-1-(2-methoxyethoxy)benzene (245 mg, yield 92%).

[0317] 2-Bromo-4-chloro-1-(2-methoxyethoxy)benzene (133 mg, 0.5 mmol), 2-thiopheneboronic acid (70 mg, 0.55 mmol), bis(triphenylphosphine)palladium(II) dichloride (35 mg, 0.05 mmol), and potassium carbonate (104 mg, 0.75 mmol) were dissolved in ethylene glycol dimethyl ether (4 mL) and water (1 mL), and the reaction was carried out at 90 °C for 2 h. After the reaction was monitored by TLC to be complete, the reaction solution was extracted with ethyl acetate. The organic phase was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After evaporation of the organic phase, the product PA014 was obtained by column chromatography purification with a yield of 45%.

[0318] 1 H NMR (500 MHz, Chloroform-d) δ 7.63 (d, J = 2.6 Hz, 1H), 7.53 (dd, J = 3.7, 1.2 Hz, 1H), 7.35 (dd, J = 5.1, 1.2 Hz, 1H), 7.17 (dd, J = 8.7, 2.6 Hz, 1H), 7.09 (dd, J = 5.1, 3.7 Hz, 1H), 6.91 (d, J = 8.8 Hz, 1H), 4.20 (dd, J = 5.6, 4.2 Hz, 2H), 3.95–3.75 (m, 2H), 3.46 (s, 3H).

[0319] Example 1-15. Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-3-(thiophen-2-yl)propanamide (PA015)

[0320] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 3-(thiophen-2-yl)propanoic acid, and finally compound PA015 was obtained with a yield of 74%.

[0321] 1 H NMR (500 MHz, Chloroform-d) δ 8.48 (d, J = 2.6 Hz, 1H), 8.19 (s, 1H), 7.13 (dd, J = 5.1, 1.2 Hz, 1H), 6.97 (dd, J = 8.6, 2.6 Hz, 1H), 6.92 (dd, J = 5.1, 3.4 Hz, 1H), 6.88–6.83 (m, 2H), 4.18–4.03 (m, 2H), 3.74–3.62 (m, 2H), 3.43 (s, 3H), 3.32–3.21 (m, 2H), 2.75 (dd, J = 8.3, 6.8 Hz, 2H).

[0322] Example 1-16. Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-4-(thiophen-2-yl)butanamide (PA016)

[0323] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 4-(thiophen-2-yl)butyric acid, and finally compound PA016 was obtained with a yield of 75%.

[0324] 1 H NMR (500 MHz, Chloroform-d) δ 8.48 (d, J = 2.5 Hz, 1H), 8.14 (s, 1H), 7.13 (dd, J = 5.2, 1.2 Hz, 1H), 6.96 (dd, J = 8.7, 2.6 Hz, 1H), 6.92 (dd, J = 5.2, 3.3 Hz, 1H), 6.85 (d, J = 8.7 Hz, 1H), 6.84–6.81 (m, 1H), 4.19–4.03 (m, 2H), 3.73–3.65 (m, 2H), 3.39 (s, 3H), 3.02–2.91 (m, 2H), 2.51–2.36 (m, 2H), 2.11 (p, J = 7.4 Hz, 2H).

[0325] Preparation of Example 1-17, N-([[1,1'-Biphenyl]-3-yl)-2-(thiophen-2-yl)acetamide (PA017)

[0326] Using a method similar to the preparation of compound PA002, 2-carboxythiophene was replaced with 2-(thiophen-2-yl)acetic acid, and finally compound PA017 was obtained with a yield of 74%.

[0327] 1 H NMR (500 MHz, Chloroform-d) δ 7.64 (t, J = 1.9 Hz, 1H), 7.59–7.52 (m, 2H), 7.50–7.39 (m, 3H), 7.39–7.29 (m, 5H), 7.06 (d, J = 3.9 Hz, 2H), 3.98 (s, 2H).

[0328] Preparation of Example 1-18, N-(5-Chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-sulfonamide (PA018)

[0329] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with thiophene-2-sulfonyl chloride, and finally compound PA018 was obtained with a yield of 72%.

[0330] 11H NMR (500 MHz, Chloroform-d) δ 7.86 (s, 1H), 7.63 (d, J = 2.5 Hz, 1H), 7.55 (dd, J = 3.8, 1.3 Hz, 1H), 7.52 (dd, J = 5.0, 1.3 Hz, 1H), 7.03–6.97 (m, 2H), 6.78 (d, J = 8.7 Hz, 1H), 3.97–3.92 (m, 2H), 3.63–3.58 (m, 2H), 3.46 (s, 3H).

[0331] Example 1-19. Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-1-hydroxy-1H-benzo[d][1,2,3]triazole-6-carboxamide (PA019)

[0332] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 1-hydroxy-1H-benzo[d][1,2,3]triazole-6-carboxylic acid, and finally compound PA019 was obtained with a yield of 53%.

[0333] 1 1H NMR (500 MHz, DMSO-d 6 ) δ 14.02 (s, 1H), 9.75 (s, 1H), 8.30 (s, 1H), 8.14 (d, J = 8.5 Hz, 1H), 7.99 (d, J = 2.2 Hz, 1H), 7.92 (d, J = 8.4 Hz, 1H), 7.27–7.18 (m, 2H), 4.24–4.18 (m, 2H), 3.70–3.65 (m, 2H), 3.28 (s, 3H).

[0334] Example 1-20. Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)furan-2-carboxamide (PA020)

[0335] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with furoic acid, and finally compound PA020 was obtained with a yield of 81%.

[0336] 1 1H NMR (500 MHz, Chloroform-d) δ 8.91 (s, 1H), 8.56 (d, J = 2.5 Hz, 1H), 7.54–7.50 (m, 1H), 7.24 (d, J = 3.5 Hz, 1H), 7.02 (dd, J = 8.7, 2.6 Hz, 1H), 6.87 (d, J = 8.7 Hz, 1H), 6.59–6.54 (m, 1H), 4.25–4.19 (m, 2H), 3.79–3.75 (m, 2H), 3.46 (s, 3H).

[0337] Example 1 - 21: Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-1H-pyrrole-2-carboxamide (PA021)

[0338] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with pyrrole-2-carboxylic acid, and finally compound PA021 was obtained with a yield of 83%.

[0339] 1 H NMR (500 MHz, Chloroform-d) δ 9.50 (s, 1H), 8.57 (s, 1H), 8.53 (d, J = 2.5 Hz, 1H), 7.07–6.96 (m, 2H), 6.88 (d, J = 8.6 Hz, 1H), 6.79–6.74 (m, 1H), 6.33–6.27 (m, 1H), 4.23–4.17 (m, 2H), 3.77–3.71 (m, 2H), 3.45 (s, 3H).

[0340] Example 1 - 22: Preparation of 5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxylic acid (PA022)

[0341] Dissolve 5-chloro-2-(2-methoxyethoxy)aniline (202 mg, 1 mmol), methyl 5-carboxy-2-thiophenecarboxylate (205 mg, 1.1 mmol), N,N-diisopropylethylamine (388 mg, 3 mmol) and HATU (418 mg, 1.1 mmol) in 3 mL of N,N-dimethylformamide, and stir the reaction at room temperature for 6 h. After TLC detection showed that the reaction was complete, the reaction solution was extracted with ethyl acetate. The organic phase was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After evaporating the organic phase, the compound methyl 5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxylate (315 mg, yield 85%) was obtained by column chromatography purification.

[0342] Dissolve methyl 5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxylate (315 mg, 0.85 mmol) and LiOH·H 2 O (357 mg, 8.5 mmol) in methanol / water (8 mL / 2 mL), and react at 80 °C for 2 h. After TLC detection showed that the reaction was complete, the organic solvent was removed by rotary evaporation. The aqueous phase was adjusted to acidic with dilute hydrochloric acid, filtered by suction, and the filter cake was placed in a vacuum drying oven for drying to obtain the product PA022 (280 mg, yield 92%).

[0343] 11H NMR (500 MHz, DMSO-d 6 ) δ 13.54 (s, 1H), 9.74 (s, 1H), 7.92 (d, J = 3.9 Hz, 1H), 7.82 (d, J = 2.3 Hz, 1H), 7.78 (d, J = 3.9 Hz, 1H), 7.24 (dd, J = 8.8, 2.4 Hz, 1H), 7.19 (d, J = 8.8 Hz, 1H), 4.21–4.16 (m, 2H), 3.70–3.64 (m, 2H), 3.28 (s, 3H).

[0344] Example 1-23, N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(3-hydroxypropyl)thiophene-2,5-dicarboxamide (PA023) Preparation

[0345] Dissolve 5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxylic acid (356 mg, 1 mmol), 3-amino-1-propanol (83 mg, 1.1 mmol), N,N-diisopropylethylamine (388 mg, 3 mmol) and HATU (418 mg, 1.1 mmol) in 3 mL of N,N-dimethylformamide, stir the reaction at room temperature for 6 h. After TLC detection showed the reaction was complete, extract the reaction solution with ethyl acetate. Wash the organic phase twice with water, once with saturated brine, and dry over anhydrous sodium sulfate. After evaporating the organic phase to dryness, purify the product PA023 (320 mg, 77%) by column chromatography.

[0346] 1 1H NMR (500 MHz, Chloroform-d) δ 8.79 (s, 1H), 8.52 (d, J = 2.6 Hz, 1H), 7.60 (d, J = 4.0 Hz, 1H), 7.51 (d, J = 4.0 Hz, 1H), 7.04 (dd, J = 8.6, 2.6 Hz, 1H), 6.91 (d, J = 8.7 Hz, 1H), 6.73–6.66 (m, 1H), 4.24–4.18 (m, 2H), 3.78 (d, J = 6.0 Hz, 2H), 3.75–3.70 (m, 2H), 3.64 (q, J = 6.0 Hz, 2H), 3.42 (s, 3H), 2.64–2.54 (m, 1H), 1.84 (p, J = 5.7 Hz, 2H).

[0347] Example 1-24, N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(4-hydroxybutyl)thiophene-2,5-dicarboxamide (PA024) Preparation

[0348] Using a method similar to that for preparing compound PA023, 3-amino-1-propanol was replaced with 4-amino-1-butanol, and finally compound PA024 was obtained with a yield of 81%.

[0349] 1 H NMR(500MHz,Chloroform-d)δ8.77(s,1H),8.51(d,J=2.6Hz,1H),7.59(d,J=3.9Hz,1H),7.52(d,J=3.9Hz,1H),7.03(dd,J=8.6,2.5Hz,1H),6.90(d,J=8.7Hz,1H),6.84–6.81(m,1H),4.23–4.18(m,2H),3.77–3.71(m,4H),3.53–3.45(m,2H),3.42(s,3H),2.07–1.94(m,1H),1.81–1.65(m,4H).

[0350] Examples 1-25, N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 Preparation of N-(6-hydroxyhexyl)thiophene-2,5-dicarboxamide (PA025)

[0351] Using a method similar to that for preparing compound PA023, 3-amino-1-propanol was replaced with 6-amino-1-hexanol, and finally compound PA025 was obtained with a yield of 75%.

[0352] 1 H NMR(500MHz,Chloroform-d)δ8.79(s,1H),8.52(d,J=2.6Hz,1H),7.60(dd,J=5.2,3.8Hz,1H),7.50(d,J=3.9Hz,1H),7.44(d,J=2.0Hz,1H),7.08–7.01(m,1H),6.92(dd,J=8.7,3.9Hz,1H),4.34(t,J=6.5Hz,1H),4.24–4.18(m,2H),3.75–3.70(m,2H),3.69–3.61(m,2H),3.46(dd,J=14.6,7.1Hz,4H),3.42(d,J=2.3Hz,3H),1.71–1.59(m,2H),1.46–1.40(m,4H).

[0353] Examples 1-26, Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-nitrothiophene-2-carboxamide (PA026)

[0354] Dissolve 5-chloro-2-(2-methoxyethoxy)aniline (202 mg, 1 mmol), 5-nitrothiophene-2-carboxylic acid (190 mg, 1.1 mmol), N,N-diisopropylethylamine (388 mg, 3 mmol) and HATU (418 mg, 1.1 mmol) in 3 mL of N,N-dimethylformamide, and stir the reaction at room temperature for 6 h. After the reaction is completed by TLC detection, extract the reaction solution with ethyl acetate. Wash the organic phase twice with water, once with saturated brine, and dry over anhydrous sodium sulfate. After evaporating the organic phase to dryness, separate and purify the product PA026 (273 mg, yield 77%) by column chromatography.

[0355] 1 1H NMR (500 MHz, Chloroform-d) δ 8.99 (s, 1H), 8.66 (s, 1H), 8.56 (d, J = 2.6 Hz, 1H), 7.52 (d, J = 4.2 Hz, 1H), 7.02 (dd, J = 8.7, 2.6 Hz, 1H), 6.91 (d, J = 8.6 Hz, 1H), 6.82 (d, J = 4.1 Hz, 1H), 4.31 (s, 2H), 4.26–4.20 (m, 2H), 3.78–3.73 (m, 2H), 3.46 (s, 3H).

[0356] Preparation of Example 1-27, 5-amino-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA027)

[0357] Dissolve N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-nitrothiophene-2-carboxamide (1.78 g, 5 mmol) in methanol (30 mL), add palladium on carbon, and introduce hydrogen gas. React at room temperature for 2 h. After the reaction is completed by TLC detection, filter through diatomaceous earth, and concentrate the filtrate to dryness to obtain the product PA027 (800 mg, yield 94%).

[0358] 1 1H NMR (500 MHz, Chloroform-d) δ 8.53–8.48 (m, 2H), 7.27 (d, J = 4.1 Hz, 1H), 6.95 (dd, J = 8.7, 2.6 Hz, 1H), 6.86 (d, J = 8.6 Hz, 1H), 6.11 (d, J = 4.0 Hz, 1H), 4.30 (s, 2H), 4.21–4.08 (m, 2H), 3.74–3.69 (m, 2H), 3.42 (s, 3H).

[0359] Preparation of Example 1-28, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(2-chloroacetamido)thiophene-2-carboxamide (PA028)

[0360] Dissolve 5-amino-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (326 mg, 1 mmol), chloroacetyl chloride (124 mg, 1.1 mmol), and triethylamine (303 mg, 3 mmol) in dichloromethane (10 mL), and stir the reaction at room temperature for 6 h. After detecting the completion of the reaction by TLC, extract the reaction solution with dichloromethane. Wash the organic phase twice with water, once with saturated brine, and dry over anhydrous sodium sulfate. After evaporating the organic phase, separate and purify the product PA028 (297 mg, yield 74%) by column chromatography.

[0361] 1 1H NMR (500 MHz, Chloroform-d) δ 8.99 (s, 1H), 8.66 (s, 1H), 8.56 (d, J = 2.6 Hz, 1H), 7.52 (d, J = 4.2 Hz, 1H), 7.02 (dd, J = 8.7, 2.6 Hz, 1H), 6.91 (d, J = 8.6 Hz, 1H), 6.82 (d, J = 4.1 Hz, 1H), 4.31 (s, 2H), 4.26–4.20 (m, 2H), 3.78–3.73 (m, 2H), 3.46 (s, 3H).

[0362] Example 1-29, N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -cyclopropylthiophene-2,5-dicarboxamide (PA029) preparation

[0363] Using a method similar to the preparation of compound PA023, replace 3-amino-1-propanol with cyclopropylamine, and finally obtain compound PA029 with a yield of 68%.

[0364] 1 1H NMR (500 MHz, Chloroform-d) δ 8.78 (s, 1H), 8.52 (d, J = 2.5 Hz, 1H), 7.59 (d, J = 3.9 Hz, 1H), 7.48 (s, 1H), 7.03 (dd, J = 8.7, 2.6 Hz, 1H), 6.91 (d, J = 8.7 Hz, 1H), 6.16 (s, 1H), 4.23–4.18 (m, 2H), 3.75–3.70 (m, 2H), 3.42 (s, 3H), 2.89 (tq, J = 7.1, 3.6 Hz, 1H), 0.93–0.83 (m, 2H), 0.70–0.63 (m, 2H).

[0365] Examples 1 - 30, N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 Preparation of -(2-hydroxyethyl)thiophene-2,5-dicarboxamide (PA030)

[0366] Using a method similar to the preparation of compound PA023, 3-amino-1-propanol was replaced with ethanolamine, and finally compound PA030 was obtained with a yield of 72%.

[0367] 1 H NMR (500 MHz, Chloroform-d) δ 8.80 (s, 1H), 8.52 (d, J = 2.6 Hz, 1H), 7.60 (d, J = 4.0 Hz, 1H), 7.52 (d, J = 3.9 Hz, 1H), 7.04 (dd, J = 8.7, 2.5 Hz, 1H), 6.91 (d, J = 8.6 Hz, 1H), 6.53 (s, 1H), 4.24–4.18 (m, 2H), 3.88–3.84 (m, 2H), 3.75–3.70 (m, 2H), 3.67–3.60 (m, 2H), 3.42 (s, 3H), 2.27–2.23 (m, 1H).

[0368] Examples 1 - 31, N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 Preparation of -(5-(5-((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)pentyl)thiophene-2,5-dicarboxamide (PA031)

[0369] Using a method similar to the preparation of compound PA023, tert-butyl (5-(5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxamido)pentyl)carbamate was prepared.

[0370] Dissolve tert-butyl (5-(5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxamido)pentyl)carbamate (270 mg, 0.5 mmol) in dichloromethane (10 mL), add hydrochloric acid / dioxane solution (3 mL), and stir at room temperature for 2 h. After TLC detection showed complete reaction, the organic solvent was removed by rotary evaporation to obtain compound N 2 -(5-aminopentyl)-N 5 -(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2,5-dicarboxamide (210 mg, yield 95%).

[0371] Dissolve N2 -(5-Aminopentyl)-N 5 -(5-Chloro-2-(2-methoxyethoxy)phenyl)thiophene-2,5-dicarboxamide (210 mg, 0.48 mmol), (+)-biotin-N-succinimidyl ester (180 mg, 0.53 mmol), and triethylamine (146 mg, 1.44 mmol) were dissolved in dichloromethane (10 mL), and the reaction mixture was stirred at room temperature for 2 h. After the reaction was completed as monitored by TLC, the reaction mixture was extracted with dichloromethane. The organic phase was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After evaporation of the organic phase, the product PA031 (253 mg, yield 80%) was obtained by column chromatography purification.

[0372] 1 H NMR (500 MHz, DMSO-d 6 ) δ 9.56 (s, 1H), 8.65 (s, 1H), 7.90–7.84 (m, 2H), 7.80–7.72 (m, 2H), 7.27–7.14 (m, 2H), 6.38 (d, J = 31.4 Hz, 2H), 4.29 (t, J = 6.5 Hz, 1H), 4.23–4.16 (m, 2H), 4.11 (d, J = 7.8 Hz, 1H), 3.71–3.65 (m, 2H), 3.29 (s, 3H), 3.26–3.20 (m, 2H), 3.10 (d, J = 8.9 Hz, 1H), 3.05–2.99 (m, 2H), 2.81 (dd, J = 12.3, 4.8 Hz, 1H), 2.59–2.54 (m, 2H), 2.04 (t, J = 7.0 Hz, 2H), 1.60 (s, 1H), 1.55–1.38 (m, 6H), 1.35–1.25 (m, 4H).

[0373] Examples 1-32, N 2 -(5-Chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(2-(2-Hydroxyethoxy)ethyl)thiophene-2,5-dicarboxamide (PA032) Preparation

[0374] Using a method similar to the preparation of compound PA023, 3-amino-1-propanol was replaced with diethanolamine, and finally compound PA032 was obtained with a yield of 72%.

[0375] 11H NMR (500 MHz, Chloroform-d) δ 8.78 (s, 1H), 8.52 (d, J = 2.5 Hz, 1H), 7.60 (d, J = 3.9 Hz, 1H), 7.54 (d, J = 3.9 Hz, 1H), 7.03 (dd, J = 8.6, 2.5 Hz, 1H), 6.91 (d, J = 8.6 Hz, 1H), 6.69 (s, 1H), 4.24–4.18 (m, 2H), 3.81–3.78 (m, 2H), 3.76–3.62 (m, 8H), 3.42 (s, 3H), 2.17–2.06 (m, 1H).

[0376] Example 1 - 33, Preparation of 5-(2-Aminoacetamido)-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA033)

[0377] Dissolve 5-Amino-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (326 mg, 1 mmol), BOC-glycine (193 mg, 1.1 mmol), N,N-diisopropylethylamine (388 mg, 3 mmol) and HATU (418 mg, 1.1 mmol) in 3 mL of N,N-dimethylformamide, and stir the reaction at room temperature for 6 h. After the reaction is complete by TLC detection, extract the reaction solution with ethyl acetate. The organic phase is washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After evaporating the organic phase, the compound tert-butyl (2-(((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-2-oxoethyl)carbamate (386 mg, yield 80%) is obtained by column chromatography separation and purification.

[0378] Dissolve tert-butyl (2-(((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-2-oxoethyl)carbamate (386 mg, 0.8 mmol) in dichloromethane (10 mL), add hydrochloric acid / dioxane solution (3 mL), and stir the reaction at room temperature for 2 h. After the reaction is complete by TLC detection, remove the organic solvent by rotary evaporation to obtain the product PA033 (257 mg, yield 84%).

[0379] 11H NMR (500 MHz, Chloroform-d) δ 8.51 (s, 1H), 8.50 (s, 1H), 7.27 (d, J = 4.0 Hz, 1H), 6.96 (dd, J = 8.6, 2.6 Hz, 1H), 6.87 (d, J = 8.5 Hz, 1H), 6.12 (d, J = 4.0 Hz, 1H), 4.27 (s, 2H), 4.21–4.16 (m, 2H), 3.74–3.69 (m, 2H), 3.42 (s, 3H).

[0380] Preparation of Examples 1 - 34, N-(2-chloro-5-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA034)

[0381] Using a method similar to the preparation of compound PA006, 4-chloro-2-nitrophenol was replaced with 2-chloro-5-nitrophenol to prepare 2-chloro-5-(2-methoxyethoxy)aniline.

[0382] Dissolve 2-chloro-5-(2-methoxyethoxy)aniline (202 mg, 1 mmol), thiophene-2-carboxylic acid (141 mg, 1.1 mmol), N,N-diisopropylethylamine (388 mg, 3 mmol) and HATU (418 mg, 1.1 mmol) in 3 mL of N,N-dimethylformamide, and stir the reaction at room temperature for 6 hours. After TLC detection showed that the reaction was complete, the reaction solution was extracted with ethyl acetate. The organic phase was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After evaporating the organic phase, the product PA034 (222 mg, yield 71%) was obtained by column chromatography separation and purification.

[0383] 1 1H NMR (500 MHz, Chloroform-d) δ 8.31 (s, 1H), 8.23 (d, J = 3.0 Hz, 1H), 7.65 (dd, J = 3.7, 1.2 Hz, 1H), 7.59 (dd, J = 5.0, 1.1 Hz, 1H), 7.28 (s, 1H), 7.16 (dd, J = 5.0, 3.7 Hz, 1H), 6.69 (dd, J = 8.9, 3.0 Hz, 1H), 4.18–4.13 (m, 2H), 3.78–3.68 (m, 2H), 3.45 (s, 3H).

[0384] Preparation of Examples 1 - 35, N-(4-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA035)

[0385] Using a method similar to that for preparing compound PA034, replace 2-chloro-5-(2-methoxyethoxy)aniline with 4-(2-methoxyethoxy)aniline, and finally obtain compound PA035 with a yield of 68%.

[0386] 1 H NMR(500MHz,Chloroform-d)δ7.60(d,J=3.0Hz,1H),7.57(s,1H),7.53(d,J=4.9Hz,1H),7.50(d,J=8.9Hz,1H),7.45(d,J=8.7Hz,1H),7.15–7.10(m,1H),6.93(d,J=8.9Hz,1H),6.84(d,J=8.7Hz,1H),4.15–4.10(m,2H),3.78–3.73(m,2H),3.46(s,3H).

[0387] Preparation of Examples 1 - 36, N-([1,1'-biphenyl]-3-yl)thiophene-3-carboxamide (PA036)

[0388] Using a method similar to that for preparing compound PA002, replace 2-thiophenecarboxylic acid with 3-thiophenecarboxylic acid, and finally obtain compound PA036 with a yield of 74%.

[0389] 1 H NMR(500MHz,Chloroform-d)δ8.00(dd,J=3.0,1.3Hz,1H),7.89–7.86(m,1H),7.75(s,1H),7.61(ddd,J=6.2,3.5,1.3Hz,3H),7.51(dd,J=5.1,1.3Hz,1H),7.47–7.32(m,6H).

[0390] Preparation of Examples 1 - 37, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(5-((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)hexanamido)thiophene-2-carboxamide (PA037)

[0391] Using a method similar to that for preparing compound PA031, replace 5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxylic acid with 5-amino-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide, and finally obtain compound PA037 with a yield of 74%.

[0392] 1 H NMR(500MHz,DMSO-d6 ) δ 11.55 (s, 1H), 9.12 (s, 1H), 8.00 (d, J = 2.1 Hz, 1H), 7.78–7.72 (m, 1H), 7.65 (d, J = 4.2 Hz, 1H), 7.19–7.12 (m, 2H), 6.70 (d, J = 4.2 Hz, 1H), 6.41 (s, 1H), 6.35 (s, 1H), 4.29 (dd, J = 7.7, 5.1 Hz, 1H), 4.22–4.17 (m, 2H), 4.15–4.09 (m, 1H), 3.71–3.66 (m, 2H), 3.31 (s, 3H), 3.12–3.06 (m, 1H), 3.02 (q, J = 6.6 Hz, 2H), 2.81 (dd, J = 12.4, 5.1 Hz, 1H), 2.57 (d, J = 12.4 Hz, 1H), 2.38 (t, J = 7.5 Hz, 2H), 2.04 (t, J = 7.4 Hz, 2H), 1.65–1.55 (m, 3H), 1.55–1.33 (m, 5H), 1.35–1.21 (m, 5H).

[0393] Preparation of Example 1-38, N-(4-(2-methoxyethyl)phenyl)thiophene-2-carboxamide (PA038)

[0394] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 4-(2-methoxyethyl)aniline, and finally compound PA038 was obtained with a yield of 72%.

[0395] 1 H NMR (500 MHz, Chloroform-d) δ 7.66 (s, 1H), 7.61 (dd, J = 3.7, 1.2 Hz, 1H), 7.56–7.50 (m, 3H), 7.24–7.19 (m, 2H), 7.12 (dd, J = 5.0, 3.7 Hz, 1H), 3.59 (t, J = 7.0 Hz, 2H), 3.36 (s, 3H), 2.87 (t, J = 7.0 Hz, 2H).

[0396] Preparation of Example 1-39, N-(3-(2-methoxyethyl)phenyl)thiophene-2-carboxamide (PA039)

[0397] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 3-(2-methoxyethyl)aniline, and finally compound PA039 was obtained with a yield of 72%.

[0398] 11H NMR (500 MHz, Chloroform-d) δ 7.72 (s, 1H), 7.62 (dd, J = 3.7, 0.9 Hz, 1H), 7.54 (dd, J = 5.0, 1.0 Hz, 1H), 7.51 (s, 1H), 7.48–7.44 (m, 1H), 7.29 (d, J = 7.8 Hz, 1H), 7.12 (dd, J = 4.9, 3.8 Hz, 1H), 7.01 (d, J = 7.6 Hz, 1H), 3.62 (t, J = 7.0 Hz, 2H), 3.36 (s, 3H), 2.89 (t, J = 7.0 Hz, 2H).

[0399] Preparation of Examples 1 - 40, N-(3-(1-methyl-1H-pyrazol-4-yl)phenyl)thiophene-2-carboxamide (PA040)

[0400] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 3-bromoaniline to prepare N-(3-bromophenyl)thiophene-2-carboxamide.

[0401] N-(3-Bromophenyl)thiophene-2-carboxamide (282 mg, 1 mmol), 1-methylpyrazole-4-boronic acid pinacol ester (312 mg, 1.5 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (73 mg, 0.1 mmol), and sodium carbonate (318 mg, 3 mmol) were dissolved in ethylene glycol dimethyl ether / water (6 mL / 3 mL), and the reaction was carried out at 90 °C for 6 h. After the reaction was completed by TLC detection, the reaction solution was extracted with ethyl acetate. The organic phase was washed twice with water and once with saturated brine, and then dried over anhydrous sodium sulfate. After evaporating the organic phase, the product PA040 (191 mg, yield 67%) was obtained by column chromatography separation and purification.

[0402] 1 1H NMR (500 MHz, Chloroform-d) δ 7.87 (s, 1H), 7.78 (s, 1H), 7.71 (s, 1H), 7.67–7.62 (m, 2H), 7.56 (dd, J = 5.0, 1.0 Hz, 1H), 7.40–7.31 (m, 2H), 7.29–7.23 (m, 1H), 7.14 (dd, J = 4.9, 3.8 Hz, 1H), 3.94 (s, 3H).

[0403] Preparation of Examples 1 - 41, N-(3-(1H-pyrazol-4-yl)phenyl)thiophene-2-carboxamide (PA041)

[0404] Using a method similar to the preparation of compound PA040, 1-methylpyrazole-4-boronic acid pinacol ester was replaced with 4-pyrazoleboronic acid pinacol ester, and finally compound PA039 was obtained with a yield of 68%.

[0405] 1 H NMR(500MHz,DMSO-d 6 )δ12.98(s,1H),10.23(s,1H),8.06(d,J=3.6Hz,1H),7.92–7.84(m,3H),7.56(d,J=7.2Hz,2H),7.38–7.25(m,2H),7.26–7.21(m,1H).

[0406] Preparation of Examples 1 - 42, N-(3-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA042)

[0407] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 3-(2-methoxyethoxy)aniline, and finally compound PA042 was obtained with a yield of 71%.

[0408] 1 H NMR(500MHz,Chloroform-d)δ7.67(s,1H),7.63–7.59(m,1H),7.55(dd,J=5.0,0.9Hz,1H),7.46–7.41(m,1H),7.28–7.20(m,1H),7.13(dd,J=4.9,3.8Hz,1H),7.08–7.03(m,1H),6.74(dd,J=8.3,1.9Hz,1H),4.18–4.13(m,2H),3.78–3.73(m,2H),3.45(s,3H).

[0409] Preparation of Examples 1 - 43, N-(2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA043)

[0410] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 2-(2-methoxyethoxy)aniline, and finally compound PA043 was obtained with a yield of 71%.

[0411] 11H NMR (500 MHz, Chloroform-d) δ 8.77 (s, 1H), 8.51–8.44 (m, 1H), 7.66 (d, J = 3.7 Hz, 1H), 7.54 (d, J = 5.0 Hz, 1H), 7.15–7.11 (m, 1H), 7.08–7.02 (m, 2H), 6.98 (dd, J = 6.0, 3.5 Hz, 1H), 4.23 (dd, J = 5.3, 3.7 Hz, 2H), 3.75 (dd, J = 5.2, 3.9 Hz, 2H), 3.43 (s, 3H).

[0412] Example 1-44. Preparation of N-(3-(1-(2-hydroxyethyl)-1H-pyrazol-4-yl)phenyl)thiophene-2-carboxamide (PA044)

[0413] Using a method similar to the preparation of compound PA040, replacing 1-methylpyrazole-4-boronic acid pinacol ester with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-ethanol, the compound PA044 was finally obtained with a yield of 65%.

[0414] 1 1H NMR (500 MHz, Chloroform-d) δ 7.87 (s, 1H), 7.80 (s, 1H), 7.76 (s, 1H), 7.73 (s, 1H), 7.65 (d, J = 3.6 Hz, 1H), 7.56 (d, J = 5.0 Hz, 1H), 7.39–7.31 (m, 2H), 7.28–7.23 (m, 1H), 7.14 (dd, J = 4.9, 3.8 Hz, 1H), 4.34–4.24 (m, 2H), 4.14–3.96 (m, 2H), 3.12–2.97 (m, 1H).

[0415] Example 1-45. Preparation of N-([1,1'-biphenyl]-3-yl)-5-(6-(5-((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentaamido)hexanamido)thiophene-2-carboxamide (PA045)

[0416] Using a method similar to the preparation of compound PA037, replacing 5-chloro-2-(2-methoxyethoxy)aniline with 3-aminobiphenyl, the compound PA045 was finally obtained with a yield of 58%.

[0417] 1 1H NMR (500 MHz, DMSO-d 6)δ11.45(s,1H),10.08(s,1H),8.03(s,1H),7.83(d,J=4.1Hz,1H),7.74(d,J=6.2Hz,2H),7.64(d,J=7.2Hz,2H),7.53–7.46(m,2H),7.45–7.34(m,3H),6.69(d,J=4.1Hz,1H),6.38(d,J=31.2Hz,2H),4.31–4.25(m,1H),4.15–4.08(m,1H),4.03(q,J=7.1Hz,1H),3.09(d,J=3.4Hz,1H),3.05–2.99(m,2H),2.80(dd,J=12.4,4.9Hz,1H),2.59–2.54(m,2H),2.41–2.34(m,2H),2.04(t,J=7.3Hz,2H),1.65–1.57(m,4H),1.55–1.36(m,6H).

[0418] Examples 1 - 46, N 2 -([1,1'-Biphenyl]-3-yl)-N 5 -(5-(5-(((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)pentyl)thiophene-2,5-dicarboxamide (PA046) Preparation

[0419] Using a method similar to the preparation of compound PA031, replacing 5-chloro-2-(2-methoxyethoxy)aniline with 3-aminobiphenyl, the compound PA046 was finally obtained with a yield of 60%.

[0420] 1 H NMR(500MHz,DMSO-d 6)δ 10.41 (s, 1H), 8.64 (s, 1H), 8.05 (s, 1H), 8.02–7.92 (m, 2H), 7.79 (d, J = 3.9 Hz, 1H), 7.75 (d, J = 8.7 Hz, 2H), 7.65 (d, J = 7.3 Hz, 2H), 7.54–7.47 (m, 2H), 7.46–7.36 (m, 2H), 6.38 (d, J = 32.4 Hz, 2H), 4.31–4.26 (m, 1H), 4.12 (s, 1H), 3.24 (d, J = 6.0 Hz, 2H), 3.05 (dd, J = 22.4, 8.1 Hz, 4H), 2.80 (dd, J = 12.3, 4.9 Hz, 1H), 2.57 (d, J = 12.3 Hz, 2H), 2.04 (t, J = 7.2 Hz, 2H), 1.55–1.38 (m, 6H), 1.35–1.26 (m, 4H).

[0421] Examples 1 - 47, N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(((2R,3R,4S,5R)-3,4,5,6-tetrahydroxy-1-oxohexan-2-yl)thiophene-2,5-dicarboxamide (PA047) Preparation

[0422] Using a method similar to the preparation of compound PA023, replacing 3-amino-1-propanol with D-glucosamine, the compound PA024 was finally obtained with a yield of 81%.

[0423] 1 H NMR (500 MHz, Chloroform-d) δ 8.81 (s, 1H), 8.48–8.42 (m, 1H), 7.55 (d, J = 4.0 Hz, 1H), 7.45 (d, J = 3.9 Hz, 1H), 7.07 (ddd, J = 7.0, 4.3, 2.0 Hz, 2H), 7.01–6.96 (m, 1H), 6.38 (s, 1H), 4.61 (dt, J = 13.6, 6.6 Hz, 1H), 4.26–4.21 (m, 2H), 3.97–3.89 (m, 4H), 3.87–3.79 (m, 5H), 3.76–3.71 (m, 2H), 3.42 (s, 3H).

[0424] Examples 1 - 48, Preparation of N-(1-(2-hydroxyethyl)-1H-pyrazol-4-yl)thiophene-2-carboxamide (PA048)

[0425] Using a method similar to that for preparing compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 2-(4-amino-1H-pyrazol-1-yl)ethanol, and finally compound PA048 was obtained with a yield of 71%.

[0426] 1 H NMR(500MHz,Chloroform-d)δ8.06(s,1H),7.60(d,J=2.9Hz,2H),7.56–7.53(m,2H),7.16–7.11(m,1H),4.27–4.21(m,2H),4.04–3.99(m,2H).

[0427] Example 1-49, Preparation of N-(5-methyl-1H-pyrazol-3-yl)thiophene-2-carboxamide (PA049)

[0428] Using a method similar to that for preparing compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 3-amino-5-methylpyrazole, and finally compound PA049 was obtained with a yield of 68%.

[0429] 1 H NMR(500MHz,Chloroform-d)δ8.35(dd,J=3.9,1.3Hz,1H),7.73(dd,J=5.0,1.3Hz,1H),7.15(dd,J=5.0,4.0Hz,1H),5.61(s,2H),5.30(s,1H),2.23(s,3H).

[0430] Example 1-50, Preparation of N-(1-methyl-1H-pyrazol-4-yl)thiophene-2-carboxamide (PA050)

[0431] Using a method similar to that for preparing compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 1-methyl-4-aminopyrazole, and finally compound PA050 was obtained with a yield of 64%.

[0432] 1 H NMR(500MHz,Chloroform-d)δ7.98(s,1H),7.95(s,1H),7.64–7.59(m,1H),7.52(dd,J=5.0,1.0Hz,1H),7.47(s,1H),7.10(dd,J=4.9,3.8Hz,1H),3.87(s,3H).

[0433] Example 1-51, Preparation of N-([1,1'-biphenyl]-3-yl)-1H-1,2,4-triazole-5-carboxamide (PA051)

[0434] Using a method similar to that for preparing compound PA002, 2-thiophenecarboxylic acid was replaced with 1H-1,2,4-triazole-3-carboxylic acid, and finally compound PA051 was obtained with a yield of 64%.

[0435] 1 H NMR(500MHz,DMSO-d 6 )δ14.64(s,1H),10.44(s,1H),8.80(s,1H),8.17(s,1H),7.88(d,J=7.4Hz,1H),7.69–7.63(m,2H),7.54–7.36(m,5H).

[0436] Preparation of Examples 1 - 52, N-([1,1'-biphenyl]-3-yl)thiazole-2-carboxamide (PA052)

[0437] Using a method similar to that for preparing compound PA002, 2-thiophenecarboxylic acid was replaced with thiazole-2-carboxylic acid, and finally compound PA052 was obtained with a yield of 70%.

[0438] 1 H NMR(500MHz,Chloroform-d)δ9.18(s,1H),8.01–7.96(m,1H),7.94(d,J=3.1Hz,1H),7.70(ddd,J=7.9,2.0,1.1Hz,1H),7.65(d,J=3.1Hz,1H),7.64–7.61(m,2H),7.49–7.40(m,4H),7.39–7.34(m,1H).

[0439] Preparation of Examples 1 - 53, N-([1,1'-biphenyl]-3-yl)thiazole-4-carboxamide (PA053)

[0440] Using a method similar to that for preparing compound PA002, 2-thiophenecarboxylic acid was replaced with thiazole-4-carboxylic acid, and finally compound PA053 was obtained with a yield of 64%.

[0441] 1 H NMR(500MHz,Chloroform-d)δ9.34(s,1H),8.83(d,J=2.1Hz,1H),8.30(d,J=2.1Hz,1H),8.00–7.96(m,1H),7.75–7.69(m,1H),7.66–7.61(m,2H),7.51–7.42(m,3H),7.41–7.33(m,2H).

[0442] Preparation of Example 1-54, N-([1,1'-biphenyl]-3-yl)-1H-pyrazole-5-carboxamide (PA054)

[0443] Using a method similar to the preparation of compound PA002, replace 2-thiophenecarboxylic acid with pyrazole-3-carboxylic acid, and finally obtain compound PA054 with a yield of 71%.

[0444] 1 H NMR (500 MHz, Chloroform-d) δ 10.28 (s, 1H), 8.79 (s, 1H), 7.95 (s, 1H), 7.69 (d, J = 8.0 Hz, 1H), 7.66 (d, J = 2.4 Hz, 1H), 7.63 (d, J = 7.4 Hz, 2H), 7.47–7.41 (m, 3H), 7.36 (dd, J = 12.4, 6.6 Hz, 2H), 7.00 (d, J = 2.3 Hz, 1H).

[0445] Preparation of Example 1-55, N-([1,1'-biphenyl]-3-yl)thiazole-5-carboxamide (PA055)

[0446] Using a method similar to the preparation of compound PA002, replace 2-thiophenecarboxylic acid with thiazole-5-carboxylic acid, and finally obtain compound PA055 with a yield of 61%.

[0447] 1 H NMR (500 MHz, Chloroform-d) δ 8.96 (s, 1H), 8.40 (s, 1H), 7.88–7.84 (m, 1H), 7.80 (s, 1H), 7.64–7.56 (m, 3H), 7.49–7.40 (m, 4H), 7.39–7.33 (m, 1H).

[0448] Preparation of Example 1-56, N-([1,1'-biphenyl]-3-yl)-1-hydroxy-1H-1,2,3-triazole-4-carboxamide (PA056)

[0449] Using a method similar to the preparation of compound PA002, replace 2-thiophenecarboxylic acid with 1-hydroxy-1H-1,2,3-triazole-4-carboxylic acid, and finally obtain compound PA056 with a yield of 78%.

[0450] 1 H NMR (500 MHz, DMSO-d 6)δ9.98(s,1H),8.15(s,1H),7.84(d,J=7.6Hz,1H),7.65(d,J=7.3Hz,2H),7.54(s,1H),7.51–7.44(m,2H),7.42–7.30(m,4H).

[0451] Examples 1 - 57, Preparation of N-([1,1'-biphenyl]-3-yl)isothiazole-5-carboxamide (PA057)

[0452] Using a method similar to the preparation of compound PA002, replacing 2-thiophenecarboxylic acid with 1-hydroxy-1H-1,2,3-triazole-4-carboxylic acid, the compound PA057 was finally obtained with a yield of 70%.

[0453] 1 H NMR(500MHz,Chloroform-d)δ8.56(d,J=1.5Hz,1H),7.86(s,1H),7.79(s,1H),7.64(d,J=1.5Hz,1H),7.63–7.56(m,3H),7.49–7.41(m,4H),7.40–7.34(m,1H).

[0454] Examples 1 - 58, Preparation of N-(thiophen-2-yl)-[1,1'-biphenyl]-3-carboxamide (PA058)

[0455] Thiophenamine (99 mg, 1 mmol), 3-aminobiphenyl (186 mg, 1.1 mmol), N,N-diisopropylethylamine (388 mg, 3 mmol) and HATU (418 mg, 1.1 mmol) were dissolved in 3 mL of N,N-dimethylformamide and stirred at room temperature for 6 h. After the reaction was detected to be complete by TLC, the reaction solution was extracted with ethyl acetate. The organic phase was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After evaporating the organic phase, the product PA058 (181 mg, yield 65%) was obtained by column chromatography separation and purification.

[0456] 1 H NMR(500MHz,Chloroform-d)δ8.75(dd,J=4.5,1.3Hz,1H),8.55–8.51(m,1H),8.48(dd,J=8.4,1.3Hz,1H),8.33–8.26(m,1H),8.02–7.97(m,1H),7.73–7.63(m,4H),7.53–7.45(m,3H),7.42(dd,J=8.4,6.4Hz,1H).

[0457] Preparation of Example 1-59, N-([1,1'-Biphenyl]-3-yl)isothiazole-5-carboxamide (PA059)

[0458] Using a method similar to the preparation of compound PA002, 2-thiophenecarboxylic acid was replaced with 5-isothiazolecarboxylic acid, and finally compound PA059 was obtained with a yield of 71%.

[0459] 1 H NMR (500 MHz, Chloroform-d) δ 8.56 (d, J = 1.5 Hz, 1H), 7.86 (s, 1H), 7.79 (s, 1H), 7.64 (d, J = 1.5 Hz, 1H), 7.63–7.56 (m, 3H), 7.49–7.41 (m, 4H), 7.40–7.34 (m, 1H).

[0460] Preparation of Example 1-60, 1-([1,1'-Biphenyl]-3-yl)-3-(thiophen-2-yl)urea (PA060)

[0461] Dissolve thiopheneamine (99 mg, 1 mmol), phenyl chloroformate (235 mg, 1.5 mmol), and potassium carbonate (414 mg, 3 mmol) in toluene / water (10 mL / 5 mL), and react at room temperature for 5 h. After TLC detection showed that the reaction was complete, the reaction solution was extracted with ethyl acetate. The organic phase was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After evaporating the organic phase, the compound phenyl thiophen-2-ylcarbamate (210 mg, yield 96%) was obtained by column chromatography separation and purification.

[0462] Dissolve phenyl thiophen-2-ylcarbamate (210 mg, 0.96 mmol) and 3-aminobiphenyl (178 mg, 1.06 mmol) in N,N-dimethylformamide (10 mL), and react at 80 °C for 2 h. After TLC detection showed that the reaction was complete, the reaction solution was extracted with ethyl acetate. The organic phase was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After evaporating the organic phase, the product PA060 was obtained by column chromatography separation and purification with a yield of 70%.

[0463] 1 H NMR (500 MHz, DMSO-d 6 ) δ 8.84 (s, 1H), 7.82 (d, J = 1.7 Hz, 1H), 7.66–7.60 (m, 3H), 7.51–7.45 (m, 3H), 7.45–7.38 (m, 3H), 7.37 (d, J = 7.7 Hz, 2H), 7.30–7.24 (m, 1H).

[0464] Preparation of Example 1-61, N-([1,1'-Biphenyl]-3-ylmethyl)thiophene-2-carboxamide (PA061)

[0465] Using a method similar to the preparation of compound PA002, 3-aminobiphenyl was replaced with 3-phenylbenzylamine, and finally compound PA061 was obtained with a yield of 71%.

[0466] 1 H NMR (500 MHz, Chloroform-d) δ 7.60–7.55 (m, 3H), 7.52 (dd, J = 8.2, 5.6 Hz, 2H), 7.48 (d, J = 5.0 Hz, 1H), 7.46–7.39 (m, 3H), 7.39–7.32 (m, 2H), 7.09–7.04 (m, 1H), 6.34 (s, 1H), 4.69 (d, J = 5.7 Hz, 2H).

[0467] Preparation of Example 1-62, N-(5-Chloro-2-(2-methoxyethoxy)phenyl)thiazole-5-carboxamide (PA062)

[0468] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with thiazole-5-carboxylic acid, and finally compound PA062 was obtained with a yield of 74%.

[0469] 1 H NMR (500 MHz, Chloroform-d) δ 8.96 (s, 1H), 8.82 (s, 1H), 8.50 (d, J = 2.5 Hz, 1H), 8.40 (s, 1H), 7.05 (dd, J = 8.7, 2.5 Hz, 1H), 6.93 (d, J = 8.7 Hz, 1H), 4.24–4.17 (m, 2H), 3.76–3.70 (m, 2H), 3.44 (s, 3H).

[0470] Preparation of Example 1-63, N-(4-Chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA063)

[0471] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 4-chloro-6-(2-methoxyethoxy)aniline, and finally compound PA063 was obtained with a yield of 71%.

[0472] 11H NMR (500 MHz, Chloroform-d) δ 8.64 (s, 1H), 8.42 (d, J = 8.7 Hz, 1H), 7.66–7.61 (m, 1H), 7.58–7.53 (m, 1H), 7.13 (dd, J = 4.9, 3.8 Hz, 1H), 7.03 (dd, J = 8.7, 2.2 Hz, 1H), 6.96 (d, J = 2.2 Hz, 1H), 4.25–4.19 (m, 2H), 3.78–3.72 (m, 2H), 3.43 (s, 3H).

[0473] Example 1 - 64, Preparation of N-(3-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA064)

[0474] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 3-chloro-6-(2-methoxyethoxy)aniline, and finally compound PA064 was obtained with a yield of 70%.

[0475] 1 1H NMR (500 MHz, Chloroform-d) δ 9.21 (s, 1H), 8.46 (p, J = 3.9 Hz, 1H), 7.75 (dd, J = 3.7, 1.1 Hz, 1H), 7.57 (dd, J = 5.0, 1.1 Hz, 1H), 7.13 (dd, J = 4.9, 3.8 Hz, 1H), 7.09 (d, J = 4.5 Hz, 2H), 4.31–4.26 (m, 2H), 3.72–3.67 (m, 2H), 3.31 (s, 3H).

[0476] Example 1 - 65, Preparation of 4-bromo-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA065)

[0477] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 4-bromothiophene-2-carboxylic acid, and finally compound PA065 was obtained with a yield of 70%.

[0478] 1 1H NMR (500 MHz, Chloroform-d) δ 8.96 (s, 1H), 8.82 (s, 1H), 8.50 (d, J = 2.5 Hz, 1H), 8.40 (s, 1H), 7.05 (dd, J = 8.7, 2.5 Hz, 1H), 6.93 (d, J = 8.7 Hz, 1H), 4.24–4.17 (m, 2H), 3.76–3.70 (m, 2H), 3.44 (s, 3H).

[0479] Example 1-66. Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-1-methyl-1H-pyrazole-3-carboxamide (PA066)

[0480] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 1-methylpyrazole-3-carboxylic acid, and finally compound PA066 was obtained with a yield of 73%.

[0481] 1 H NMR (500 MHz, Chloroform-d) δ 9.41 (s, 1H), 8.60 (d, J = 2.5 Hz, 1H), 7.40 (d, J = 2.3 Hz, 1H), 6.98 (dd, J = 8.7, 2.6 Hz, 1H), 6.87 (d, J = 2.3 Hz, 1H), 6.83 (d, J = 8.7 Hz, 1H), 4.23–4.18 (m, 2H), 3.96 (s, 3H), 3.85–3.80 (m, 2H), 3.51 (s, 3H).

[0482] Example 1-67. Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(methylsulfonyl)thiophene-2-carboxamide (PA067)

[0483] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 5-methylsulfonylthiophene-2-carboxylic acid, and finally compound PA067 was obtained with a yield of 75%.

[0484] 1 H NMR (500 MHz, Chloroform-d) δ 8.89 (s, 1H), 8.50 (d, J = 2.5 Hz, 1H), 7.71 (d, J = 4.0 Hz, 1H), 7.62 (d, J = 4.0 Hz, 1H), 7.07 (dd, J = 8.7, 2.5 Hz, 1H), 6.94 (d, J = 8.7 Hz, 1H), 4.24–4.19 (m, 2H), 3.75–3.69 (m, 2H), 3.41 (s, 3H), 3.23 (s, 3H).

[0485] Example 1-68. Preparation of 3-bromo-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA068)

[0486] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 3-bromothiophene-2-carboxylic acid, and finally compound PA068 was obtained with a yield of 70%.

[0487] 11H NMR (500 MHz, Chloroform-d) δ 9.70 (s, 1H), 8.61 (d, J = 2.4 Hz, 1H), 7.52 (dd, J = 5.3, 1.1 Hz, 1H), 7.10 (dd, J = 5.3, 1.1 Hz, 1H), 7.03 (dt, J = 8.7, 1.8 Hz, 1H), 6.83 (d, J = 8.7 Hz, 1H), 4.27–4.16 (m, 2H), 3.83–3.72 (m, 2H), 3.42 (d, J = 1.1 Hz, 3H).

[0488] Example 1-69. Preparation of N-(5-chloro-2-(2-hydroxyethyl)phenyl)thiophene-2-carboxamide (PA069)

[0489] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 2-(2-amino-4-chlorophenyl)ethan-1-ol, and finally compound PA069 was obtained with a yield of 72%.

[0490] 1 1H NMR (500 MHz, DMSO-d 6 ) δ 9.31 (s, 1H), 7.89–7.84 (m, 2H), 7.67 (d, J = 2.2 Hz, 1H), 7.26–7.21 (m, 1H), 7.03 (d, J = 8.3 Hz, 1H), 6.95 (dd, J = 8.4, 2.2 Hz, 1H), 4.17–4.11 (m, 2H), 3.69–3.63 (m, 2H).

[0491] Example 1-70. Preparation of N-(5-chloro-2-(pentoxy)phenyl)thiophene-2-carboxamide (PA070)

[0492] Using a method similar to the preparation of compound PA006, 2-bromoethyl methyl ether was replaced with 1-bromopentane to prepare 4-chloro-2-(pentoxy)aniline.

[0493] 4-Chloro-2-(pentoxy)aniline (214 mg, 1 mmol), 2-carboxythiophene (141 mg, 1.1 mmol), N,N-diisopropylethylamine (388 mg, 3 mmol) and HATU (418 mg, 1.1 mmol) were dissolved in 3 mL of N,N-dimethylformamide and stirred at room temperature for 6 hours. After the reaction was completed as detected by TLC, the reaction solution was extracted with ethyl acetate. The organic phase was washed twice with water, once with saturated brine, and dried over anhydrous sodium sulfate. After evaporating the organic phase, the product PA070 (233 mg, yield 72%) was obtained by column chromatography separation and purification.

[0494] 1 1H NMR (500 MHz, Chloroform-d) δ 8.54 (d, J = 2.5 Hz, 1H), 8.45 (s, 1H), 7.60 (dd, J = 3.7, 1.0 Hz, 1H), 7.56 (dd, J = 5.0, 1.0 Hz, 1H), 7.14 (dd, J = 4.9, 3.8 Hz, 1H), 7.01 (dd, J = 8.7, 2.5 Hz, 1H), 6.81 (d, J = 8.7 Hz, 1H), 4.06 (t, J = 6.5 Hz, 2H), 1.88 (dt, J = 14.5, 6.5 Hz, 2H), 1.53–1.48 (m, 2H), 1.46–1.40 (m, 2H), 0.96 (t, J = 7.2 Hz, 3H).

[0495] Example 1-71, Preparation of N-(5-chloro-2-ethoxyphenyl)thiophene-2-carboxamide (PA071)

[0496] Using a method similar to the preparation of compound PA070, replacing n-pentyl bromide with ethyl bromide, the compound PA071 was finally obtained with a yield of 70%.

[0497] 1 1H NMR (500 MHz, Chloroform-d) δ 8.53 (d, J = 2.5 Hz, 1H), 8.45 (s, 1H), 7.57 (dd, J = 17.6, 4.2 Hz, 2H), 7.17–7.12 (m, 1H), 7.00 (dd, J = 8.7, 2.5 Hz, 1H), 6.80 (d, J = 8.7 Hz, 1H), 4.13 (q, J = 7.0 Hz, 2H), 1.50 (t, J = 7.0 Hz, 3H).

[0498] Example 1-72, Preparation of 4-chloro-2-(thiophene-2-carboxamido)phenylacetic acid (PA072)

[0499] Using a method similar to the preparation of compound PA070, replacing n-pentyl bromide with acetic anhydride, the compound PA072 was finally obtained with a yield of 81%.

[0500] 1 1H NMR (500 MHz, Chloroform-d) δ 8.35 (s, 1H), 8.03 (d, J = 3.0 Hz, 1H), 7.77–7.72 (m, 1H), 7.28 (s, 1H), 7.25–7.22 (m, 1H), 7.21 (d, J = 8.7 Hz, 1H), 7.13 (d, J = 8.2 Hz, 1H), 2.16 (s, 3H).

[0501] Preparation of Example 1-73, N-(2-(2-Methoxyethoxy)-5-methylphenyl)thiophene-2-carboxamide (PA073)

[0502] Using a method similar to the preparation of compound PA034, replace 2-chloro-5-nitrophenol with 2-nitro-4-methylphenol, and finally obtain compound PA073 with a yield of 73%.

[0503] 1 H NMR (500 MHz, Chloroform-d) δ 7.40–7.36 (m, 2H), 7.32 (dd, J = 5.0, 1.1 Hz, 1H), 7.08–7.03 (m, 3H), 6.84 (dd, J = 4.9, 3.9 Hz, 1H), 4.64 (s, 3H), 2.44 (t, J = 2.4 Hz, 2H), 2.24 (t, J = 2.5 Hz, 2H), 1.57 (s, 3H).

[0504] Preparation of Example 1-74, N-(2-Butoxy-5-chlorophenyl)thiophene-2-carboxamide (PA074)

[0505] Using a method similar to the preparation of compound PA070, replace n-pentyl bromide with n-butyl bromide, and finally obtain compound PA074 with a yield of 81%.

[0506] 1 H NMR (500 MHz, Chloroform-d) δ 8.54 (d, J = 2.5 Hz, 1H), 8.45 (s, 1H), 7.60–7.58 (m, 1H), 7.56 (d, J = 5.0 Hz, 1H), 7.15 (dd, J = 4.9, 3.8 Hz, 1H), 7.01 (dd, J = 8.7, 2.5 Hz, 1H), 6.81 (d, J = 8.7 Hz, 1H), 4.07 (t, J = 6.5 Hz, 2H), 1.86 (dt, J = 14.4, 6.5 Hz, 2H), 1.57–1.52 (m, 2H), 1.03 (t, J = 7.4 Hz, 3H).

[0507] Preparation of Example 1-75, N-(5-Chloro-2-propoxyphenyl)thiophene-2-carboxamide (PA075)

[0508] Using a method similar to the preparation of compound PA070, replace n-pentyl bromide with n-propyl bromide, and finally obtain compound PA075 with a yield of 81%.

[0509] 11H NMR (500 MHz, Chloroform-d) δ 8.56 (d, J = 2.5 Hz, 1H), 8.49 (s, 1H), 7.62 (dd, J = 3.7, 1.0 Hz, 1H), 7.58 (dd, J = 5.0, 1.0 Hz, 1H), 7.17 (dd, J = 4.9, 3.8 Hz, 1H), 7.03 (dd, J = 8.7, 2.6 Hz, 1H), 6.83 (d, J = 8.7 Hz, 1H), 4.05 (t, J = 6.5 Hz, 2H), 1.93 (h, J = 7.3 Hz, 2H), 1.14 (t, J = 7.4 Hz, 3H).

[0510] Example 1-76. Preparation of N-(5-chloro-2-(3-methoxypropoxy)phenyl)thiophene-2-carboxamide (PA076)

[0511] Using a method similar to the preparation of compound PA070, n-pentyl bromide was replaced with 1-bromo-3-methoxypropane, and finally compound PA076 was obtained with a yield of 74%.

[0512] 1 1H NMR (500 MHz, Chloroform-d) δ 8.53 (d, J = 2.5 Hz, 1H), 8.52 (s, 1H), 7.62 (dd, J = 3.7, 1.1 Hz, 1H), 7.56 (dd, J = 5.0, 1.1 Hz, 1H), 7.14 (dd, J = 5.0, 3.8 Hz, 1H), 7.01 (dd, J = 8.7, 2.6 Hz, 1H), 6.83 (d, J = 8.7 Hz, 1H), 4.17 (t, J = 6.2 Hz, 2H), 3.60 (t, J = 5.9 Hz, 2H), 3.33 (s, 3H), 2.13 (p, J = 6.1 Hz, 2H).

[0513] Example 1-77. Preparation of N-(5-chloro-2-(2-ethoxyethoxy)phenyl)thiophene-2-carboxamide (PA077)

[0514] Using a method similar to the preparation of compound PA070, n-pentyl bromide was replaced with 2-bromoethyl ethyl ether, and finally compound PA077 was obtained with a yield of 71%.

[0515] 11H NMR (500 MHz, Chloroform-d) δ 8.68 (s, 1H), 8.54 (d, J = 2.5 Hz, 1H), 7.65 (dd, J = 3.7, 1.1 Hz, 1H), 7.56 (dd, J = 5.0, 1.1 Hz, 1H), 7.13 (dd, J = 5.0, 3.8 Hz, 1H), 7.01 (dd, J = 8.7, 2.6 Hz, 1H), 6.88 (d, J = 8.7 Hz, 1H), 4.24–4.19 (m, 2H), 3.82–3.74 (m, 2H), 3.59 (q, J = 7.0 Hz, 2H), 1.21 (t, J = 7.0 Hz, 3H).

[0516] Example 1-78. Preparation of N-(5-chloro-2-(2-hydroxyethoxy)phenyl)thiophene-2-carboxamide (PA078)

[0517] Using a method similar to the preparation of compound PA070, n-pentyl bromide was replaced with 2-bromoethanol, and finally compound PA078 was obtained with a yield of 76%.

[0518] 1 1H NMR (500 MHz, Chloroform-d) δ 8.61 (s, 1H), 8.49 (d, J = 2.4 Hz, 1H), 7.65 (dd, J = 3.7, 1.2 Hz, 1H), 7.54 (dd, J = 5.0, 1.2 Hz, 1H), 7.12 (dd, J = 5.0, 3.7 Hz, 1H), 7.02 (dd, J = 8.7, 2.5 Hz, 1H), 6.88 (d, J = 8.8 Hz, 1H), 4.24–4.14 (m, 2H), 4.06–3.96 (m, 2H), 1.95 (s, 1H).

[0519] Example 1-79. Preparation of N-(5-chloro-2-(2,2,2-trifluoroethoxy)phenyl)thiophene-2-carboxamide (PA079)

[0520] Using a method similar to the preparation of compound PA070, n-pentyl bromide was replaced with 2-bromo-1,1,1-trifluoroethane, and finally compound PA079 was obtained with a yield of 76%.

[0521] 11H NMR (500 MHz, Chloroform-d) δ 8.65 (s, 1H), 8.52 (d, J = 2.5 Hz, 1H), 7.65 (dd, J = 3.7, 1.1 Hz, 1H), 7.56 (dd, J = 5.0, 1.1 Hz, 1H), 7.12 (dd, J = 5.0, 3.8 Hz, 1H), 7.01 (dd, J = 8.7, 2.6 Hz, 1H), 6.88 (d, J = 8.8 Hz, 1H), 4.78 (q, J = 8.5 Hz, 2H).

[0522] Example 1-80. Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiazole-2-carboxamide (PA080)

[0523] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with thiazole-2-carboxylic acid, and finally compound PA080 was obtained with a yield of 68%.

[0524] 1 1H NMR (500 MHz, Chloroform-d) δ 8.96 (s, 1H), 8.82 (s, 1H), 8.50 (d, J = 2.5 Hz, 1H), 8.40 (s, 1H), 7.05 (dd, J = 8.7, 2.5 Hz, 1H), 6.93 (d, J = 8.7 Hz, 1H), 4.24–4.17 (m, 2H), 3.76–3.70 (m, 2H), 3.44 (s, 3H).

[0525] Example 1-81. Preparation of N-(5-chloro-2-(heptyloxy)phenyl)thiophene-2-carboxamide (PA081)

[0526] Using a method similar to the preparation of compound PA070, n-pentyl bromide was replaced with 1-bromoheptane, and finally compound PA081 was obtained with a yield of 63%.

[0527] 11H NMR (500 MHz, Chloroform-d) δ 8.53 (d, J = 2.5 Hz, 1H), 8.45 (s, 1H), 7.59 (dd, J = 3.7, 1.1 Hz, 1H), 7.56 (dd, J = 5.0, 1.1 Hz, 1H), 7.14 (dd, J = 4.9, 3.8 Hz, 1H), 7.00 (dd, J = 8.7, 2.6 Hz, 1H), 6.80 (d, J = 8.7 Hz, 1H), 4.06 (t, J = 6.4 Hz, 2H), 1.87 (dt, J = 14.4, 6.5 Hz, 2H), 1.55–1.45 (m, 2H), 1.43–1.36 (m, 2H), 1.35–1.29 (m, 4H), 0.95–0.87 (m, 3H).

[0528] Example 1 - 82. Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-6-ethynylnicotinamide (PA082)

[0529] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 6-ethynylnicotinic acid, and finally compound PA082 was obtained with a yield of 71%.

[0530] 1 1H NMR (500 MHz, Chloroform-d) δ 9.23 (dd, J = 2.3, 0.9 Hz, 1H), 9.08 (s, 1H), 8.56 (d, J = 2.5 Hz, 1H), 8.42 (dd, J = 8.1, 2.2 Hz, 1H), 7.85 (dd, J = 8.1, 0.8 Hz, 1H), 7.09 (dd, J = 8.7, 2.5 Hz, 1H), 6.97 (d, J = 8.6 Hz, 1H), 4.27–4.17 (m, 2H), 4.12 (q, J = 7.1 Hz, 1H), 3.79–3.66 (m, 2H), 3.39 (s, 3H).

[0531] Example 1 - 83. Preparation of methyl 4-chloro-2-(thiophene-2-carboxamido)benzoate (PA083)

[0532] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with methyl 2-amino-4-chlorophenol ester, and finally compound PA083 was obtained with a yield of 76%.

[0533] 11H NMR (500 MHz, Chloroform-d) δ 8.35 (s, 1H), 8.03 (d, J = 3.0 Hz, 1H), 7.77–7.72 (m, 1H), 7.28 (s, 1H), 7.25–7.22 (m, 1H), 7.21 (d, J = 8.7 Hz, 1H), 7.13 (d, J = 8.2 Hz, 1H), 3.96 (s, 3H).

[0534] Example 1-84, Preparation of N-(2-(2-Methoxyethyl)phenyl)thiophene-2-carboxamide (PA084)

[0535] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 2-(2-methoxyethyl)aniline, and finally compound PA084 was obtained with a yield of 74%.

[0536] 1 1H NMR (500 MHz, Chloroform-d) δ 9.77 (s, 1H), 8.01 (d, J = 8.0 Hz, 1H), 7.64 (dd, J = 3.7, 1.0 Hz, 1H), 7.53 (dd, J = 5.0, 1.1 Hz, 1H), 7.33–7.26 (m, 1H), 7.18 (dd, J = 7.6, 1.5 Hz, 1H), 7.15–7.07 (m, 2H), 3.77–3.71 (m, 2H), 3.45 (s, 3H), 2.95–2.89 (m, 2H).

[0537] Example 1-85, Preparation of N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-(cyclopropanecarboxamido)thiophene-2-carboxamide (PA085)

[0538] Using a method similar to the preparation of compound PA028, chloroacetyl chloride was replaced with cyclopropylcarbonyl chloride, and finally compound PA085 was obtained with a yield of 74%.

[0539] 11H NMR (500 MHz, Chloroform-d) δ 8.57 (s, 1H), 8.53 (d, J = 2.5 Hz, 1H), 8.35 (s, 1H), 7.46 (d, J = 4.1 Hz, 1H), 6.98 (dd, J = 8.7, 2.5 Hz, 1H), 6.87 (d, J = 8.6 Hz, 1H), 6.65 (d, J = 4.1 Hz, 1H), 4.22–4.15 (m, 2H), 3.75–3.70 (m, 2H), 3.43 (s, 3H), 1.71–1.65 (m, 1H), 1.20–1.14 (m, 2H), 0.95–0.92 (m, 2H).

[0540] Example 1-86. Preparation of N-(5-chloro-2-(prop-2-yn-1-yloxy)phenyl)thiophene-2-carboxamide (PA086)

[0541] Using a method similar to the preparation of compound PA070, bromopentane was replaced with 3-bromopropyne, and finally compound PA086 was obtained with a yield of 74%.

[0542] 1 1H NMR (500 MHz, Chloroform-d) δ 7.41–7.35 (m, 2H), 7.32 (dd, J = 5.0, 1.1 Hz, 1H), 7.08–7.02 (m, 3H), 6.84 (dd, J = 4.9, 3.9 Hz, 1H), 4.64 (s, 2H), 2.44 (t, J = 2.4 Hz, 1H).

[0543] Example 1-87. Preparation of (5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carbonyl)glycine (PA087)

[0544] Using a method similar to the preparation of compound PA023, 3-amino-1-propanol was replaced with glycine, and finally compound PA087 was obtained with a yield of 69%.

[0545] 1 1H NMR (500 MHz, DMSO-d 6 ) δ 9.63 (s, 1H), 8.99–8.96 (m, 1H), 7.90 (d, J = 3.8 Hz, 1H), 7.88–7.82 (m, 2H), 7.25–7.15 (m, 2H), 4.21–4.17 (m, 2H), 3.89–3.84 (m, 2H), 3.70–3.66 (m, 2H), 3.28 (s, 3H).

[0546] Preparation of Example 1-88, N-(5-Methoxy-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA088)

[0547] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 5-methoxy-2-(2-methoxyethyl)aniline, and finally compound PA088 was obtained with a yield of 70%.

[0548] 1 H NMR (500 MHz, Chloroform-d) δ 8.89 (s, 1H), 8.21 (d, J = 3.0 Hz, 1H), 7.68 (dd, J = 3.7, 1.1 Hz, 1H), 7.55 (dd, J = 5.0, 1.1 Hz, 1H), 7.13 (dd, J = 4.9, 3.8 Hz, 1H), 6.93 (d, J = 8.9 Hz, 1H), 6.59 (dd, J = 8.9, 3.0 Hz, 1H), 4.19–4.14 (m, 2H), 3.81 (s, 3H), 3.71–3.65 (m, 2H), 3.39 (s, 3H).

[0549] Preparation of Example 1-89, N-(5-Chloro-2-(2-methoxyethoxy)phenyl)-5-methoxypyrazine-2-carboxamide (PA089)

[0550] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 5-methoxypyrazine-2-carboxylic acid, and finally compound PA089 was obtained with a yield of 70%.

[0551] 1 H NMR (500 MHz, Chloroform-d) δ 10.20 (s, 1H), 9.01 (d, J = 1.3 Hz, 1H), 8.65 (d, J = 2.5 Hz, 1H), 8.15 (d, J = 1.3 Hz, 1H), 7.03 (dd, J = 8.7, 2.6 Hz, 1H), 6.86 (d, J = 8.7 Hz, 1H), 4.25–4.20 (m, 2H), 4.07 (s, 3H), 3.82 (dd, J = 5.3, 3.9 Hz, 2H), 3.49 (s, 3H).

[0552] Preparation of Example 1-90, N-(5-Bromo-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA090)

[0553] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 5-bromo-2-(2-methoxyethoxy)aniline, and finally compound PA090 was obtained with a yield of 76%.

[0554] 1 H NMR(500MHz,Chloroform-d)δ8.72(s,1H),8.69(d,J=2.4Hz,1H),7.65(dd,J=3.7,1.0Hz,1H),7.56(dd,J=5.0,1.0Hz,1H),7.19–7.11(m,2H),6.86–6.82(m,1H),4.23–4.18(m,2H),3.76–3.71(m,2H),3.42(s,3H).

[0555] Example 1-91, Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-4-phenylthiazole-2-carboxamide (PA091)

[0556] Using a method similar to the preparation of compound PA006, 5-methoxypicolinic acid was replaced with 4-phenylthiazole-2-carboxylic acid, and finally compound PA091 was obtained with a yield of 70%.

[0557] 1 H NMR(500MHz,Chloroform-d)δ9.98(s,1H),8.59(d,J=2.5Hz,1H),8.01–7.95(m,2H),7.78(s,1H),7.50–7.43(m,2H),7.43–7.36(m,1H),7.06(dd,J=8.7,2.5Hz,1H),6.88(d,J=8.7Hz,1H),4.28–4.22(m,2H),3.90–3.84(m,2H),3.48(s,3H).

[0558] Example 1-92, Preparation of N-(5-fluoro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA092)

[0559] Using a method similar to the preparation of compound PA034, 2-chloro-5-(2-methoxyethoxy)aniline was replaced with 5-bromo-2-(2-methoxyethoxy)aniline, and finally compound PA092 was obtained with a yield of 72%.

[0560] 11H NMR (500 MHz, Chloroform-d) δ 8.85 (s, 1H), 8.31 (dd, J = 10.7, 3.0 Hz, 1H), 7.67 (d, J = 2.8 Hz, 1H), 7.56 (d, J = 4.9 Hz, 1H), 7.16–7.11 (m, 1H), 6.93 (dd, J = 8.9, 5.0 Hz, 1H), 6.77–6.70 (m, 1H), 4.19 (dd, J = 5.3, 3.6 Hz, 2H), 3.71 (dd, J = 5.3, 3.6 Hz, 2H), 3.40 (s, 3H).

[0561] Example 1-93. Preparation of tert-butyl (5-(5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxamido)pentyl)carbamate (PA093)

[0562] Using a method similar to the preparation of compound PA023, 3-amino-1-propanol was replaced with tert-butyl N-(5-aminopentyl)carbamate, and finally compound PA093 was obtained with a yield of 76%.

[0563] 1 1H NMR (500 MHz, Chloroform-d) δ 8.78 (s, 1H), 8.53 (d, J = 2.5 Hz, 1H), 7.60 (d, J = 4.0 Hz, 1H), 7.53 (d, J = 4.0 Hz, 1H), 7.03 (dd, J = 8.7, 2.6 Hz, 1H), 6.91 (d, J = 8.6 Hz, 1H), 4.24–4.18 (m, 2H), 3.75–3.70 (m, 2H), 3.49–3.43 (m, 2H), 3.42 (s, 3H), 3.14 (d, J = 7.0 Hz, 2H), 3.00 (s, 2H), 1.66 (p, J = 7.1 Hz, 2H), 1.58–1.54 (m, 9H), 1.53 (d, J = 7.3 Hz, 2H), 1.38 (d, J = 5.1 Hz, 2H).

[0564] Example 1-94. Preparation of (5-(5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxamido)pentyl)carbamate (PA094)

[0565] Dissolve tert-butyl (5-(5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxamido)pentyl)carbamate (540 mg, 1 mmol) in 10 ml of dichloromethane, add 2 mL of trifluoroacetic acid, react at room temperature for 2 hours, evaporate the solvent, and use ethyl acetate and saturated NaHCO 3Extraction was carried out, the organic phase was rotary evaporated to dryness, and the product PA094 (412 mg, 94%) was obtained by column chromatography separation and purification.

[0566] 1 H NMR (500 MHz, Chloroform-d) δ 8.79 (s, 1H), 8.52 (d, J = 2.6 Hz, 1H), 7.63–7.57 (m, 1H), 7.52 (d, J = 3.9 Hz, 1H), 7.03 (dd, J = 8.7, 2.6 Hz, 1H), 6.91 (d, J = 8.7 Hz, 1H), 6.36–6.27 (m, 1H), 4.24–4.18 (m, 2H), 3.75–3.70 (m, 2H), 3.48–3.44 (m, 2H), 3.42 (s, 3H), 2.76–2.71 (m, 2H), 1.68–1.65 (m, 6H), 1.54–1.43 (m, 2H).

[0567] Preparation of Example 1 - 95, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(2-(2-(2-(5-(((3aR,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)ethoxy)ethoxy)acetamido)thiophene-2-carboxamide (PA095)

[0568] Using a method similar to the preparation of compound PA037, tert-butoxycarbonyl 6-aminohexanoic acid was replaced with 2-[2-(tert-butoxycarbonylamino)ethoxy]ethoxyacetic acid, and finally compound PA095 was obtained with a yield of 59%.

[0569] 1 H NMR (500 MHz, DMSO-d 6)δ11.40(s,1H),9.18(s,1H),8.01–7.97(m,2H),7.85(t,J=5.6Hz,1H),7.68(d,J=4.2Hz,1H),7.17(d,J=1.9Hz,2H),6.90(d,J=4.2Hz,1H),6.42(s,1H),6.36(s,1H),4.32–4.26(m,1H),4.24–4.18(m,4H),4.14–4.09(m,1H),3.72–3.66(m,4H),3.61(dd,J=5.7,3.5Hz,2H),3.44(t,J=5.9Hz,2H),3.31(s,3H),3.21(q,J=5.8Hz,2H),3.12–3.04(m,1H),2.81(dd,J=12.4,5.1Hz,1H),2.60–2.54(m,1H),2.07(t,J=7.4Hz,2H),1.65–1.40(m,4H),1.36–1.25(m,2H).

[0570] Examples 1 - 96, N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(2-(2-(2-(5-(((3aR,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamidoethoxy)ethoxy)ethyl)thiophene-2,5-dicarboxamide (PA096) Preparation

[0571] Using a method similar to the preparation of compound PA031, tert-butyl N-(5-aminopentyl)carbamate was replaced with tert-butyl 2-[2-(2-aminoethoxy)ethoxy]ethylcarbamate N-Boc-3,6-dioxa-1,8-octanediamine, and finally compound PA096 was obtained with a yield of 62%.

[0572] 1 H NMR(500MHz,DMSO-d 6)δ9.60(s,1H),8.84–8.77(m,1H),7.88(d,J=3.2Hz,2H),7.85–7.74(m,2H),7.27–7.17(m,2H),6.42(s,1H),6.37(s,1H),4.34–4.28(m,1H),4.22–4.18(m,2H),4.15–4.10(m,1H),3.69(t,J=4.6Hz,2H),3.56–3.51(m,4H),3.44–3.37(m,4H),3.37–3.32(m,2H),3.29(s,3H),3.18(q,J=5.9Hz,2H),3.12–3.06(m,1H),2.81(dd,J=12.4,5.1Hz,1H),2.61–2.55(m,1H),2.06(t,J=7.5Hz,2H),1.65–1.41(m,4H),1.32–1.22(m,2H).

[0573] Preparation of Example 1-97, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(thiophene-2-carboxamido)hexanamido)thiophene-2-carboxamide (PA097)

[0574] Using a method similar to the preparation of compound PA031, D-biotin was replaced with thiophene-2-carboxylic acid, and finally compound PA097 was obtained with a yield of 71%.

[0575] 1 H NMR(500MHz,Chloroform-d)δ9.51(s,1H),8.63(s,1H),8.50–8.34(m,1H),7.59–7.54(m,1H),7.45(dd,J=5.9,4.4Hz,2H),7.07(dd,J=5.0,3.7Hz,1H),7.05–6.99(m,1H),6.99–6.93(m,1H),6.72(d,J=4.1Hz,1H),6.51(t,J=5.9Hz,1H),4.22(dd,J=5.3,3.8Hz,2H),3.80–3.67(m,2H),3.44(s,3H),2.43(t,J=7.2Hz,2H),1.86–1.71(m,2H),1.47–1.19(m,4H),0.98–0.82(m,2H).

[0576] Preparation of Example 1-98, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(2-phenylacetamido)hexanamido)thiophene-2-carboxamide (PA098)

[0577] Using a method similar to the preparation of compound PA031, D-biotin was replaced with phenylacetic acid, and finally compound PA098 was obtained with a yield of 75%.

[0578] 1 H NMR (500 MHz, Chloroform-d) δ 9.41 (s, 1H), 8.61 (s, 1H), 8.49–8.38 (m, 1H), 7.46 (d, J = 4.1 Hz, 1H), 7.37–7.26 (m, 3H), 7.26–7.20 (m, 2H), 7.07–6.99 (m, 2H), 6.99–6.92 (m, 1H), 6.70 (d, J = 4.1 Hz, 1H), 4.25–4.19 (m, 2H), 3.79–3.71 (m, 2H), 3.56 (s, 2H), 3.44 (s, 3H), 3.22 (q, J = 6.8 Hz, 2H), 2.38 (t, J = 7.3 Hz, 2H), 1.73 (p, J = 7.4 Hz, 2H), 1.47 (p, J = 7.2 Hz, 2H), 0.95–0.83 (m, 2H).

[0579] Examples 1-99, N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 Preparation of N-(2-(2-(2-(2-(2-phenylacetamido)ethoxy)ethoxy)ethyl)thiophene-2,5-dicarboxamide (PA099)

[0580] Using a method similar to the preparation of compound PA098, tert-butyl N-(5-aminopentyl)carbamate was replaced with tert-butyl 2-(2-(2-aminoethoxy)ethoxy)ethylcarbamate, and finally compound PA099 was obtained with a yield of 70%.

[0581] 1 H NMR (500 MHz, Chloroform-d) δ 8.80 (s, 1H), 8.51 (d, J = 2.5 Hz, 1H), 7.61–7.54 (m, 2H), 7.37–7.25 (m, 4H), 7.25 (s, 1H), 7.04 (dd, J = 8.6, 2.6 Hz, 1H), 6.95–6.83 (m, 2H), 5.99 (s, 1H), 4.24–4.18 (m, 2H), 3.75–3.69 (m, 2H), 3.62 (d, J = 2.6 Hz, 4H), 3.57 (s, 6H), 3.53 (t, J = 5.3 Hz, 2H), 3.47–3.40 (m, 2H), 3.41 (s, 3H).

[0582] Preparation of Example 1-100, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(2-(thiophen-2-yl)acetamido)hexanamido)thiophene-2-carboxamide (PA100)

[0583] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with 2-thiopheneacetic acid, and finally obtain compound PA100 with a yield of 71%.

[0584] 1 H NMR (500 MHz, Chloroform-d) δ 9.33 (s, 1H), 8.59 (s, 1H), 8.52 (d, J = 2.5 Hz, 1H), 7.46 (d, J = 4.1 Hz, 1H), 7.23 (dd, J = 5.2, 1.2 Hz, 1H), 7.01–6.95 (m, 2H), 6.92 (d, J = 3.1 Hz, 1H), 6.87 (d, J = 8.6 Hz, 1H), 6.72 (d, J = 4.2 Hz, 1H), 5.86 (s, 1H), 4.21–4.14 (m, 2H), 3.77 (s, 2H), 3.75–3.71 (m, 2H), 3.44 (s, 3H), 3.25 (q, J = 6.7 Hz, 2H), 2.41 (t, J = 7.3 Hz, 2H), 1.75 (p, J = 7.4 Hz, 2H), 1.50 (p, J = 7.2 Hz, 2H), 0.96–0.83 (m, 2H).

[0585] Preparation of Example 1-101, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(3-(4-fluorophenyl)propanamido)hexanamido)thiophene-2-carboxamide (PA101)

[0586] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with 3-(4-fluorophenyl)propionic acid, and finally obtain compound PA101 with a yield of 75%.

[0587] 11H NMR (500 MHz, Chloroform-d) δ 9.09 (s, 1H), 8.59 (s, 1H), 8.53 (d, J = 2.5 Hz, 1H), 7.46 (d, J = 4.2 Hz, 1H), 7.16–7.09 (m, 2H), 7.01–6.91 (m, 3H), 6.88 (d, J = 8.7 Hz, 1H), 6.72 (d, J = 4.2 Hz, 1H), 5.61 (s, 1H), 4.23–4.17 (m, 2H), 3.80–3.66 (m, 2H), 3.43 (s, 3H), 3.22 (q, J = 6.7 Hz, 2H), 2.92 (t, J = 7.5 Hz, 2H), 2.43 (dt, J = 19.5, 7.4 Hz, 4H), 1.78–1.70 (m, 2H), 1.51–1.43 (m, 2H), 0.95–0.89 (m, 2H).

[0588] Preparation of Example 1-102, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(4-(thiophen-2-yl)butanamido)hexanamido)thiophene-2-carboxamide (PA102)

[0589] Using a method similar to the preparation of compound PA098, replacing phenylacetic acid with 4-(thiophen-2-yl)butyric acid, the compound PA102 was finally obtained with a yield of 69%.

[0590] 1 1H NMR (500 MHz, DMSO-d 6 ) δ 11.50 (s, 1H), 9.11 (s, 1H), 8.00 (d, J = 2.3 Hz, 1H), 7.78 (t, J = 5.6 Hz, 1H), 7.65 (d, J = 4.1 Hz, 1H), 7.29 (dd, J = 5.1, 1.2 Hz, 1H), 7.21–7.09 (m, 2H), 6.93 (dd, J = 5.1, 3.4 Hz, 1H), 6.85–6.79 (m, 1H), 6.68 (d, J = 4.1 Hz, 1H), 4.25–4.13 (m, 2H), 3.74–3.63 (m, 2H), 3.31 (s, 3H), 3.03 (q, J = 6.6 Hz, 2H), 2.76 (t, J = 7.6 Hz, 2H), 2.37 (t, J = 7.4 Hz, 2H), 2.10 (t, J = 7.4 Hz, 2H), 1.81 (t, J = 7.5 Hz, 2H), 1.68–1.53 (m, 2H), 1.41 (p, J = 7.1 Hz, 2H), 1.33–1.16 (m, 2H).

[0591] Preparation of Example 1-103, 2-chloro-N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)pyrimidine-5-carboxamide (PA103)

[0592] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with 2-chloropyrimidine-5-carboxylic acid, and finally obtain compound PA103 with a yield of 69%.

[0593] 1 H NMR (500 MHz, DMSO-d 6 ) δ 11.52 (dd, J = 12.2, 6.5 Hz, 1H), 9.24–9.02 (m, 2H), 8.90–8.65 (m, 1H), 7.99 (d, J = 7.0 Hz, 1H), 7.65 (dd, J = 9.5, 4.3 Hz, 1H), 7.16 (d, J = 3.0 Hz, 2H), 6.75–6.62 (m, 2H), 4.20 (t, J = 4.6 Hz, 2H), 3.69 (dd, J = 5.9, 3.1 Hz, 2H), 3.30 (d, J = 2.0 Hz, 3H), 2.40 (p, J = 7.2 Hz, 2H), 1.73–1.45 (m, 4H), 1.34 (td, J = 17.8, 16.4, 7.1 Hz, 2H), 1.28–1.15 (m, 2H).

[0594] Preparation of Example 1-104, 3-bromo-N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)furan-2-carboxamide (PA104)

[0595] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with 3-bromo-2-furoic acid, and finally obtain compound PA104 with a yield of 78%.

[0596] 11H NMR (500 MHz, Chloroform-d) δ 9.18 (s, 1H), 8.58 (s, 1H), 8.53 (d, J = 2.5 Hz, 1H), 7.46 (d, J = 4.2 Hz, 1H), 7.43 (d, J = 1.9 Hz, 1H), 6.98 (dd, J = 8.6, 2.5 Hz, 1H), 6.87 (d, J = 8.6 Hz, 1H), 6.73 (d, J = 4.2 Hz, 1H), 6.61 (d, J = 6.5 Hz, 1H), 6.57 (d, J = 1.9 Hz, 1H), 4.23–4.17 (m, 2H), 3.76–3.70 (m, 2H), 3.44 (s, 3H), 2.45 (t, J = 7.4 Hz, 2H), 1.82 (p, J = 7.5 Hz, 2H), 1.65 (s, 2H), 1.51–1.37 (m, 2H), 1.39–1.17 (m, 2H).

[0597] Example 1-105. Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(4-chloro-2-methylbenzamido)hexanamido)thiophene-2-carboxamide (PA105)

[0598] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with 4-chloro-2-methylbenzoic acid to finally obtain compound PA105 with a yield of 71%.

[0599] 1 1H NMR (500 MHz, Chloroform-d) δ 9.45 (s, 1H), 8.58 (s, 1H), 8.53–8.44 (m, 1H), 7.40 (d, J = 4.1 Hz, 1H), 7.23 (d, J = 8.2 Hz, 1H), 7.17 (s, 1H), 7.11 (s, 1H), 6.98 (dd, J = 8.6, 2.5 Hz, 1H), 6.87 (d, J = 8.7 Hz, 1H), 6.64 (d, J = 4.2 Hz, 1H), 6.18 (s, 1H), 4.23–4.15 (m, 2H), 3.79–3.66 (m, 2H), 3.42 (s, 3H), 2.45 (t, J = 7.3 Hz, 2H), 2.38 (s, 3H), 1.80 (d, J = 8.0 Hz, 2H), 1.66 (d, J = 7.5 Hz, 2H), 1.43 (dt, J = 21.0, 7.4 Hz, 2H), 1.38–1.19 (m, 2H).

[0600] Preparation of Example 1-106, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(3-(thiophen-2-yl)propanamido)hexanamido)thiophene-2-carboxamide (PA106)

[0601] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with (thiophen-2-yl)propanoic acid, and finally obtain compound PA106 with a yield of 70%.

[0602] 1 H NMR (500 MHz, Chloroform-d) δ 9.45 (s, 1H), 8.58 (s, 1H), 8.53–8.44 (m, 1H), 7.40 (d, J = 4.1 Hz, 1H), 7.23 (d, J = 8.2 Hz, 1H), 7.17 (s, 1H), 7.11 (s, 1H), 6.98 (dd, J = 8.6, 2.5 Hz, 1H), 6.87 (d, J = 8.7 Hz, 1H), 6.64 (d, J = 4.2 Hz, 1H), 6.18 (s, 1H), 4.23–4.15 (m, 2H), 3.79–3.66 (m, 2H), 3.42 (s, 3H), 2.45 (t, J = 7.3 Hz, 2H), 2.38 (s, 3H), 1.80 (d, J = 8.0 Hz, 2H), 1.66 (d, J = 7.5 Hz, 2H), 1.43 (dt, J = 21.0, 7.4 Hz, 2H), 1.38–1.19 (m, 2H).

[0603] Example 1-107, N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)thiophene-2,5-dicarboxamide (PA107) Preparation

[0604] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with 5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxylic acid, and finally obtain compound PA107 with a yield of 71%.

[0605] 1 H NMR (500 MHz, DMSO-d 6)δ 11.47 (s, 1H), 9.55 (s, 1H), 9.06 (s, 1H), 8.68 (d, J = 7.5 Hz, 1H), 7.96–7.82 (m, 2H), 7.77 (d, J = 4.1 Hz, 1H), 7.62 (d, J = 4.2 Hz, 1H), 7.29–7.15 (m, 2H), 7.15–7.06 (m, 2H), 7.03–6.91 (m, 1H), 6.68 (d, J = 4.1 Hz, 1H), 4.18 (q, J = 5.0 Hz, 4H), 3.68 (dt, J = 6.2, 3.0 Hz, 4H), 3.36–3.21 (m, 6H), 2.39 (q, J = 9.9, 8.6 Hz, 2H), 1.68–1.61 (m, 2H), 1.56 (t, J = 7.7 Hz, 2H), 1.41–1.31 (m, 2H), 1.28–1.14 (m, 2H).

[0606] Preparation of Example 1 - 108, N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)benzofuran-2-carboxamide (PA108)

[0607] Using a method similar to the preparation of compound PA098, replacing phenylacetic acid with benzofuran-2-carboxylic acid, the compound PA108 was finally obtained with a yield of 73%.

[0608] 1 H NMR (500 MHz, DMSO-d 6 )δ 11.47 (s, 1H), 9.55 (s, 1H), 9.06 (s, 1H), 8.68 (d, J = 7.5 Hz, 1H), 7.96–7.82 (m, 2H), 7.77 (d, J = 4.1 Hz, 1H), 7.62 (d, J = 4.2 Hz, 1H), 7.29–7.15 (m, 2H), 7.15–7.06 (m, 2H), 7.03–6.91 (m, 1H), 6.68 (d, J = 4.1 Hz, 1H), 4.18 (q, J = 5.0 Hz, 4H), 3.68 (dt, J = 6.2, 3.0 Hz, 4H), 3.36–3.21 (m, 6H), 2.39 (q, J = 9.9, 8.6 Hz, 2H), 1.68–1.61 (m, 2H), 1.56 (t, J = 7.7 Hz, 2H), 1.41–1.31 (m, 2H), 1.28–1.14 (m, 2H).

[0609] Preparation of Example 1-109, N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)thiazole-5-carboxamide (PA109)

[0610] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with thiazole-5-carboxylic acid, and finally obtain compound PA109 with a yield of 73%.

[0611] 1 H NMR (500 MHz, Chloroform-d) δ 8.88 (s, 1H), 8.84 (s, 1H), 8.62 (s, 1H), 8.50 (d, J = 2.5 Hz, 1H), 8.31 (s, 1H), 7.46 (d, J = 4.1 Hz, 1H), 6.99 (dd, J = 8.7, 2.6 Hz, 1H), 6.88 (d, J = 8.7 Hz, 1H), 6.70 (d, J = 4.1 Hz, 1H), 6.51 (s, 1H), 4.20 (d, J = 2.7 Hz, 2H), 3.78–3.69 (m, 2H), 3.52–3.45 (m, 2H), 3.43 (s, 3H), 2.47 (t, J = 7.2 Hz, 2H), 1.80 (q, J = 7.5 Hz, 2H), 1.70–1.64 (m, 4H).

[0612] Preparation of Example 1-110, N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(2-(pyridin-2-yl)acetamido)hexanamido)thiophene-2-carboxamide (PA110)

[0613] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with 2-pyridylacetic acid, and finally obtain compound PA110 with a yield of 71%.

[0614] 11H NMR (500 MHz, Chloroform-d) δ 9.33 (s, 1H), 8.58 (s, 1H), 8.57–8.50 (m, 2H), 7.67 (td, J = 7.7, 1.8 Hz, 1H), 7.48 (d, J = 4.2 Hz, 1H), 7.43 (d, J = 16.4 Hz, 1H), 7.26–7.18 (m, 2H), 6.98 (dd, J = 8.7, 2.5 Hz, 1H), 6.88 (d, J = 8.7 Hz, 1H), 6.73 (d, J = 4.1 Hz, 1H), 4.20 (d, J = 5.0 Hz, 2H), 3.75–3.72 (m, 2H), 3.72 (s, 2H), 3.44 (s, 3H), 3.28 (q, J = 6.6 Hz, 2H), 2.41 (t, J = 7.3 Hz, 2H), 1.77 (p, J = 7.4 Hz, 2H), 1.57–1.50 (m, 2H), 1.46–1.37 (m, 2H).

[0615] Example 1 - 111. Preparation of N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)-1H-pyrazole-5-carboxamide (PA111)

[0616] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with 1H-pyrazole-5-carboxylic acid to finally obtain compound PA111 with a yield of 71%.

[0617] 1 1H NMR (600 MHz, DMSO-d 6 ) δ 13.17 (s, 1H), 11.50 (s, 1H), 9.11 (s, 1H), 8.08 (t, J = 6.1 Hz, 1H), 8.00 (d, J = 2.3 Hz, 1H), 7.79 (t, J = 1.8 Hz, 1H), 7.64 (d, J = 4.2 Hz, 1H), 7.17–7.14 (m, 2H), 6.68 (d, J = 4.2 Hz, 1H), 6.60 (t, J = 2.2 Hz, 1H), 4.25–4.12 (m, 2H), 3.76–3.59 (m, 2H), 3.31 (s, 3H), 3.22 (p, J = 6.5 Hz, 2H), 2.42–2.35 (m, 2H), 1.63 (p, J = 7.5 Hz, 2H), 1.52 (p, J = 7.4 Hz, 2H), 1.32 (dd, J = 15.1, 7.5 Hz, 2H).

[0618] Example 1-112. Preparation of 5-(6-(2-(1H-tetrazol-1-yl)acetamido)hexanamido)-N-(5-chloro-2-(2-methoxyethoxy)phenyl)thiophene-2-carboxamide (PA112)

[0619] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with tetrazoleacetic acid, and finally obtain compound PA112 with a yield of 77%.

[0620] 1 H NMR (500 MHz, DMSO-d 6 ) δ 11.52 (s, 1H), 9.35 (s, 1H), 9.12 (s, 1H), 8.42 (t, J = 5.6 Hz, 1H), 7.99 (d, J = 2.2 Hz, 1H), 7.65 (d, J = 4.3 Hz, 1H), 7.28–7.05 (m, 2H), 6.69 (d, J = 4.2 Hz, 1H), 5.21 (s, 2H), 4.25–4.14 (m, 2H), 3.74–3.64 (m, 2H), 3.31 (s, 3H), 3.11 (d, J = 6.3 Hz, 2H), 2.39 (t, J = 7.5 Hz, 2H), 1.69–1.57 (m, 2H), 1.52–1.39 (m, 2H), 1.32 (tt, J = 9.5, 6.1 Hz, 2H).

[0621] Example 1-113. Preparation of N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(4-phenylbutanamido)hexanamido)thiophene-2-carboxamide (PA113)

[0622] Using a method similar to the preparation of compound PA098, replace phenylacetic acid with 4-phenylbutyric acid, and finally obtain compound PA113 with a yield of 71%.

[0623] 11H NMR (500 MHz, Chloroform-d) δ 9.54 (s, 1H), 8.68–8.44 (m, 2H), 7.45 (d, J = 4.1 Hz, 1H), 7.26 (t, J = 7.5 Hz, 2H), 7.16 (dd, J = 19.7, 7.3 Hz, 3H), 7.00–6.95 (m, 1H), 6.87 (d, J = 8.6 Hz, 1H), 6.73 (d, J = 4.1 Hz, 1H), 5.88–5.59 (m, 1H), 4.29–4.10 (m, 2H), 3.85–3.65 (m, 2H), 3.43 (d, J = 0.8 Hz, 3H), 3.24 (q, J = 6.7 Hz, 2H), 2.63 (t, J = 7.5 Hz, 2H), 2.43 (t, J = 7.3 Hz, 2H), 2.18 (t, J = 7.6 Hz, 2H), 1.96 (p, J = 7.5 Hz, 2H), 1.76 (p, J = 7.4 Hz, 2H), 1.51 (q, J = 7.2 Hz, 2H), 1.35 (p, J = 7.3, 6.9 Hz, 2H).

[0624] Summary of the preparation of the compounds shown in Examples 1-113 (PA001-113) in Table 1

[0625]

[0626]

[0627]

[0628]

[0629]

[0630]

[0631]

[0632]

[0633]

[0634]

[0635]

[0636]

[0637]

[0638]

[0639] The compounds of the present invention can be identified using a variety of assays; an initial screen identified compounds that stimulate human mesenchymal stem cells (hMSCs) to develop into chondrocyte nodules.

[0640] Example 2: Biological experiments

[0641] Example 2-1, Culturing and differentiation of cells

[0642] Human bone marrow mesenchymal stem cells (hBMSCs), umbilical cord mesenchymal stem cells (hUCMSCs), and adipose mesenchymal stem cells (hADMSCs) were selected. They were cultured in mesenchymal stem cell growth medium (MSCGM), and the 2-8th generation mesenchymal stem cells in good condition obtained by culturing could be used for experiments. The cells were sorted by fluorescence-activated cell sorting, and it was demonstrated that the positive rates for CD29, CD44, CD166, and CD105 were >98%, and the positive rate for CD45 was <0.1%.

[0643] Example 2-2, Immunocytochemical staining and quantification

[0644] To induce the chondrogenic differentiation of MSCs, 5000 cells / well were plated in a Corning 384-well plate. After 24 hours, the MSCGM was removed and replaced with 25 μL of DMEM containing 1% FBS. Then, the indicated doses of the test compounds were added to each well. The cultures were grown at 37 °C for 7 days.

[0645] To detect potential proteins promoting chondrogenic differentiation, the cells were fixed with 4% PFA for 15 minutes, permeabilized with PBS containing 0.1% triton X-100 and 0.25 g / mL collagenase 2 for 10 minutes, blocked with PBST containing 5% BSA at room temperature for 1 hour, and then incubated overnight at 4 °C with type II collagen antibody in PBS containing 1% BSA. The cells were washed 3 times with PBS, incubated with a fluorophore-conjugated secondary antibody at room temperature for 1 hour, then washed 3 times with PBS, incubated with DAPI for 3 minutes, and then washed 3 times with PBS. High-content estimation quantification was performed using ImageXpress Micro. Data analysis was carried out using a dedicated multi-wavelength cell scoring script to calculate the level of type II collagen expression stimulated by the compounds prepared in Example 1 of the present invention.

[0646] For the evaluation of compound activity, the negative control (DMSO) was used as the basal level of MSCs chondrogenic differentiation. Compounds prepared in Example 1 of the present invention that showed a staining intensity increase of 30% or more compared to the negative control were selected as active hits. Representative data are shown in Table 1 (A: staining intensity increase > 200% compared to the negative control; B: staining intensity increase 100 - 200% compared to the negative control; C: staining intensity increase 30 - 100% compared to the negative control; D: staining intensity increase < 30% compared to the negative control).

[0647] As shown in Table 2, the vast majority of compounds prepared in Example 1 of the present invention were able to significantly promote the expression of type II collagen in mesenchymal stem cells, and multiple compounds showed better activity than the positive compound KGN at a concentration of 10 μM. These results indicate that the compounds prepared in Example 1 of the present invention can induce chondrogenic differentiation of mesenchymal stem cells.

[0648] Table 2. Activity of the compounds of the present invention in inducing chondrogenesis

[0649]

[0650]

[0651] Example 3: Animal experiment

[0652] Example 3 - 1 C57 mouse surgically induced acute injury osteoarthritis model

[0653] In the OA mouse acute injury surgery, namely the medial meniscus instability surgery (DMM) model, the method causes instability of the knee joint and thus damages articular cartilage by surgically cutting the anterior cruciate ligament (ACL), medial meniscus tibial ligament (MMTL), and medial collateral ligament (MCL) of the right knee of C57 mice (number of mice in each group n = 7), resulting in the formation of OA phenotypes. To evaluate the chondroprotective ability of the test compounds, the experiment was divided into four groups: Sham sham surgery group (only the skin was incised without harming the joint ligaments), negative control (intra - articular injection of PBS), KGN administration group (KGN was dissolved in physiological saline to form a 33 μM solution for intra - articular injection), and test compound PA002 administration group (the test compound was dissolved in physiological saline to form a 33 μM solution for intra - articular injection). Three days after surgery, each group was given intra - articular injection of the drug once a week. After 6 weeks, the mice were sacrificed, the joint samples were taken, embedded and sectioned, and histological improvement was observed and statistically analyzed.

[0654] The results are as Figure 1As shown, it can be found that the compound PA002 of the present invention can significantly reverse the process of cartilage damage at a dosage of 33 μM. In terms of joint score results, PA002 shows better cartilage repair effect, and there is obvious statistical significance. In the behavioral pain measurement experiment on mice, the mice in the PA002 administration group showed good response to the pain caused by postoperative osteoarthritis, and the effect was better than that of the KGN administration group. It indicates that PA002 can well relieve the pain caused by osteoarthritis.

[0655] Example 3-2 C57 mouse articular cartilage defect osteoarthritis model

[0656] Use a 1 mL disposable sterile syringe needle (26G) to induce cartilage damage. The needle is polished in advance to make it blunt, and a 10 μL transparent pipette tip is put on the above needle, so that the tip exposes 1 mm. The needle is exposed to ultraviolet radiation for 15 minutes for sterilization before the operation. When the mice grow to 4 weeks old, the operation begins. Male C57BL / 6 mice are anesthetized with avertin (220 μL / 15 g). Wipe the hind limbs and the operating table with cotton balls dipped in 75% alcohol to disinfect, and try to reduce the pain of the mice during the operation. Use a surgical scalpel to make a skin incision and muscle layer with a length slightly less than 1 cm on the medial side of the proximal femur, open the joint capsule, move the patella to the side, and then bend the joint completely to expose the articular surface of the trochlear groove. Use the above needle to cause longitudinal cartilage damage in the patellar groove, place the tip of the needle in front of the patellar groove, and gently move it along the entire length of the femur. Confirm the penetration to the subchondral bone through the bleeding at the cartilage damage site. Suture the joint capsule and skin with mouse suture. The contralateral knee joint is not operated (sham operation control group). Place the mice on a constant temperature heating plate at 37 °C, and after waking up, put them into the mouse cage and continue to raise them in the SPF-class barrier environment for 7, 10, and 14 days. The experiment is divided into four groups (the number of mice in each group n = 7), namely the sham operation group, the PBS group, the KGN administration group, and the test compound PA002 administration group. In both administration groups, the drug is prepared into a concentration of 33 μM with normal saline for intra-articular injection. Three days after the model is established, intra-articular injection is given once a week. Four weeks later, the mice are sacrificed by cervical dislocation, the femurs are dissected, photographed with a stereomicroscope, the tissues are fixed, and then decalcified and sectioned, and safranin-fast green staining is used to observe the process of cartilage damage repair.

[0657] The results are as Figure 2 shown. It can be found that the positive compound KGN does not have a good cartilage damage repair effect, while the compound PA002 of the present invention can significantly promote cartilage repair at the cartilage defect site at a dosage of 33 μM, indicating that the compound PA002 has a good effect on cartilage damage repair.

[0658] Example 3-3 Construction of SD rat articular cartilage injury model

[0659] All surgeries were performed in a sterile environment, and the experimental instruments were sterilized. In this study, male Sprague-Dawley rats (8 weeks old) were used; the rats were anesthetized by injecting 4% chloral hydrate (1 ml / 100 g), their skin was prepared, disinfected, and draped; the skin and muscle were incised successively to expose the knee joint; a full-thickness cylindrical cartilage defect with a diameter of 2 mm and a depth of 1.5 mm was drilled using an electric drill; the animal experiments were divided into four groups (the number of mice in each group n = 7), namely the sham operation group, the PBS control group, the UC-MSCs direct injection group, and the UC-MSCs injection group after in vitro induction with 5k (10 μM). Five days after the model was established, stem cells were injected into the joint cavity; 9 weeks after the operation, the rats were sacrificed by excessive injection of 4% chloral hydrate, and all specimens were collected; after the repair period was completed, the cartilage repair status of each sample was obtained by a digital camera ( Figure 3 A). The present invention also intends to utilize a biomaterial, that is, to wrap stem cells with gelatin and fix them to the cartilage defect site for targeted repair of articular cartilage. Using the same modeling and drug administration methods as the above experiment, femoral joint samples of rats were taken 9 weeks after the operation ( Figure 3 B).

[0660] The results are as Figure 3 shown. Figure 3 A The results showed that in the PBS group without stem cell treatment, less regenerated cartilage tissue was observed in the cartilage defect area, and obvious cartilage defects could still be observed. In the experimental group directly injected with UC-MSCs, it was obvious that the articular cartilage defect site was repaired, but the joint surface was relatively rough, showing a slight phenomenon of bone hyperplasia, which was not conducive to cartilage repair. In the experimental group of UC-MSCs induced in vitro by PA002, the phenomenon of abnormal cartilage hyperplasia was weaker, and it was observed that the cartilage defect site was well repaired. It could be observed that a complete cartilage tissue was formed on the defect surface and formed a smooth surface with the surrounding original cartilage tissue. These experimental results indicate that the in vitro induction with the compound PA002 in advance can improve the in vivo cartilage repair effect of MSCs. Figure 3The results of Group B showed that obvious cartilage defects could still be observed on the joint surface in the hydrogel group without administration. In the group of undifferentiated UC-MSCs wrapped in hydrogel, a layer of hyaline cartilage-like tissue was observed to form at the site of articular cartilage defect, and the cartilage defect site could be clearly observed. It was speculated that the formed cartilage might be thinner and the subchondral bone site was not well repaired. In the experimental group induced by compound PA002, it was observed that the cartilage defect site had been completely covered with smooth tissue, and the color of the newly formed tissue was not much different from that of the surrounding normal cartilage tissue. This result indicated that compound PA002 could better promote the chondrogenic differentiation of UC-MSCs in vivo, and after UC-MSCs induced by PA002 were wrapped in hydrogel, it could better promote the repair of cartilage defects and avoid the problem of abnormal tissue hyperplasia caused by direct injection of stem cells.

[0661] The protection scope of the present invention is not limited to the above embodiments. Without departing from the spirit and scope of the inventive concept, changes and advantages that can be conceived by those skilled in the art are included in the present invention, and the appended claims are taken as the protection scope.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that, the compound is selected from: N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -cyclopropylthiophene-2,5-dicarboxamide; N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(2-hydroxyethyl)thiophene-2,5-dicarboxamide; N-([1,1'-biphenyl]-3-yl)-5-(6-(5-((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)hexanamido)thiophene-2-carboxamide; N 2 -([1,1'-Biphenyl]-3-yl)-N 5 -(5-(5-(((3aR,4R,6aS)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)pentyl)thiophene-2,5-dicarboxamide; tert-butyl (5-(5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophene-2-carboxamido)pentyl)carbamate; N-([1,1'-biphenyl]-3-yl)-2-(thiophen-2-yl)acetamide; N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(4-(thiophen-2-yl)butanamido)hexanamido)thiophene-2-carboxamide; 2-chloro-N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)pyrimidine-5-carboxamide; N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)benzofuran-2-carboxamide; N 2 -(5-chloro-2-(2-methoxyethoxy)phenyl)-N 5 -(2-(2-(2-(2-(2-phenylacetamido)ethoxy)ethoxy)ethyl)thiophene-2,5-dicarboxamide; N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(2-(thiophen-2-yl)acetamido)hexanamido)thiophene-2-carboxamide; N-(5-chloro-2-(2-methoxyethoxy)phenyl)-5-(6-(4-chloro-2-methylbenzamido)hexanamido)thiophene-2-carboxamide; N-(6-((5-((5-chloro-2-(2-methoxyethoxy)phenyl)carbamoyl)thiophen-2-yl)amino)-6-oxohexyl)thiazole-5-carboxamide.

2. A pharmaceutical composition, characterized in that, the pharmaceutical composition comprises the compound or a pharmaceutically acceptable salt thereof as claimed in claim 1 and a pharmaceutically acceptable excipient.

3. The pharmaceutical composition as claimed in claim 2, characterized in that, the pharmaceutical composition further comprises an additional compound capable of effectively treating, improving, preventing arthritis or joint injury of a mammal and / or symptoms associated with arthritis or joint injury.

4. The pharmaceutical composition as claimed in claim 3, characterized in that, the additional compound is selected from one or more of NSAIDS, analgesics, angiopoietin-like 3 protein or its chondrogenic variant, oral salmon calcitonin, iNOS inhibitor, vitamin D3, apoptosis / caspase inhibitor, collagen hydrolysate, FGF18, BMP7, avocado soybean unsaponifiables, hyaluronic acid.

5. Use of the compound or a pharmaceutically acceptable salt thereof as claimed in claim 1, or the pharmaceutical composition as claimed in any one of claims 2-4 in the preparation of a pharmaceutical composition for preventing and / or treating and / or improving arthritis or joint injury of a mammal and / or symptoms associated with arthritis or joint injury.

6. The use as claimed in claim 5, characterized in that, the arthritis is one or more of osteoarthritis, traumatic arthritis, autoimmune arthritis.

7. The application according to claim 5, wherein, the compound or its pharmaceutically acceptable salt, and the pharmaceutical composition are administered to a desired subject in a matrix and a biocompatible scaffold.

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