Benzamide compound and use thereof

Abstract

Disclosed in the present invention are a benzamide compound and a use thereof, and provided are a benzamide compound represented by formula I or a pharmaceutically acceptable salt, ester, stereoisomer, deuterated compound, or solvate thereof. Further disclosed in the present invention is a use of the compound or the pharmaceutically acceptable salt, ester, stereoisomer, deuterated compound, or solvate thereof in preparation of a drug for preventing or treating a USP7-mediated disease. The compound provided in the present invention has strong USP7 inhibitory activity and high metabolic stability, while significantly reducing hERG inhibitory activity, thereby having improved druggability.

Description

Benzamide compounds and uses thereof TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, in particular to benzamide compounds, and the preparation method and application thereof. BACKGROUND

[0002] USP7 (also known as HAUSP) is an important deubiquitinase, which is involved in cell cycle regulation, DNA repair, protein stability regulation and other biological processes in the human body through various mechanisms, and its abnormal expression is closely related to the occurrence and development of tumors. For example, USP7 can stabilize MDM2 protein in tumor cells through deubiquitination, thereby reducing the level of p53 (Cell 1998, 95, 5). Inhibition of USP7 in tumor cells can promote the degradation of MDM2 protein and increase the level of p53, thereby inhibiting tumor cell proliferation (Mol Cell 2004, 6, 879; Nature 2004, 428, 6982). USP7 can also stabilize Foxp3 protein in Treg cells, promoting the differentiation and immunosuppressive function of Treg cells. Inhibition of USP7 in Treg cells can reduce the level of Foxp3 protein, down-regulate the level and activity of Treg cells, and thus improve the immunosuppressive microenvironment of tumors (Immunity 2013, 39(2), 259; EbioMedicine 2016, 13, 99). Therefore, inhibition of USP7 is expected to become a new way to treat tumors. In addition, USP7 is also closely related to inflammatory diseases, viral infectious diseases and other diseases. Studies have shown that USP7 can stabilize key inflammatory factors such as NF-κB, promote the expression of inflammatory genes, and thus exacerbate inflammatory reactions. USP7 inhibitors can down-regulate the expression of inflammatory factors such as NF-κB, reduce tissue damage and inflammatory reactions (Cell Death and Disease 2014, 5, e1229). Another study has shown that USP7 can interact with the nuclear antigen 1 protein of Epstein-Barr virus, and assist in the carcinogenicity of Epstein-Barr virus (Biochemical Society Transactions 2004, 32, 731-732). USP7 can also enhance the DNA replication of KSHV virus through similar effects (JoV 2012, 86(12):6745-57). USP7 can also control the generation of HIV-1 by stabilizing Tat protein (Biochem J. 2017, 474(10): 1653-1668). USP7 inhibitors can synergize with proteasome inhibitors to reduce the replication ability of HIV virus and resist retroviral activity (Frontiers in Microbiology 2022, 13).

[0003] In summary, USP7 has become a potential drug development target, and its inhibitors have broad potential application prospects in anti-tumor, anti-inflammatory diseases and anti-viral diseases, but there is no USP7 inhibitor entering clinical research at present. The reported potent USP7 inhibitors have very limited structural types and various defects. For example, Gavory et al. reported N-acylpiperidinols represented by Compound 4 (CP4) as potent USP7 inhibitors (Nat Chem Biol 2018, 14(2), 118), however, the metabolic stability of CP4 and the like in human liver microsomes is poor. RAPT company reported thienopyridines represented by Compound 41 (CP41) as potent USP7 inhibitors, but the compounds have strong hERG inhibitory activity, and there is a great risk of cardiotoxic side effects (J Med Chem 2020, 63, 5398). Developing new USP7 inhibitors with good drug properties has potential clinical transformation value. SUMMARY

[0004] The first object of the present application is to provide a class of benzamide compounds, which can act as USP7 inhibitors and have excellent activity, or pharmaceutically acceptable salts, esters, stereoisomers, deuterium derivatives or solvates thereof. The second object of the present application is to provide the use of the benzamide compounds or pharmaceutically acceptable salts, esters, stereoisomers, deuterium derivatives or solvates thereof in the preparation of drugs for preventing or treating USP7-mediated diseases.

[0005] The present application provides a benzamide compound as shown in formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, deuterium derivative or solvate thereof:

[0006] wherein:

[0007] R 1 , R 2 each independently is selected from H, halogen, -CN, alkynyl, -N3, -OR 4 , -O(CO)R 4 , -SR 4 , -NR 4 R 5 , -NR 4 (CO)R 5 , -(CO)OR 4 , -(CO)NR 4 R 5 , -SF5, C 1-3 alkyl or deuterated alkyl or cycloalkyl or halogenated alkyl or halogenated cycloalkyl;

[0008] Ring A is unsubstituted or substituted with R 3a , R 3b is a 4-8 membered nitrogen-containing non-aromatic heterocyclic ring;

[0009] R 3a , R 3b are each independently selected from H, halogen, oxo (=0), oximino (=N0H), =N0R 6 , -CN, -N3, -OR 6 , -O(CO)R 6 , -SR 6 , -NR 6 R 7 , -NR 6 (CO)R 7 , -(CO)OR 6 , -(CO)NR 6 R 7 , -SO2R 6 , -SO2NR 6 R 7 , C 1-6 1-6 alkyl, C 2-6 2-6 alkenyl, C 2-6 2-6 alkynyl, 3-8 cycloalkyl or heterocycloalkyl, 5-10 aryl or heteroaryl, which alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl groups are optionally substituted with halogen, -CN, -(CO)OR 8 , -(CO)NR 8 R 9 , -NO2, -OR 8 , -NR 8 R 9 , C 1-6 1-6 alkyl, C 2-6 2-6 alkenyl, C 2-6 2-6 alkynyl, C 3-8 3-8 cycloalkyl or C 3-8 4-8 heterocycloalkyl; or R 3a , R 3b are linked together with the ring atoms to which they are attached to form a 4-7 membered ring;

[0010] M is a carbon atom or a nitrogen atom to which a substituent Z 3 , Z 4 is attached, when M is a carbon atom C 3 , Z 1 , Z 2 , Z 3 , Z 4 are each independently selected from H, C 1-3 halogenated or unhalogenated alkyl or cycloalkyl, Z 1 , Z 2, Z 3 , Z 4 any one of which can be connected to any one of the atoms contained in the other three, thereby forming together with the atoms to which it is connected a 3- to 6-membered ring;

[0011] R 4 , R 5 , R 6 , R 7 , R 8 , R 9 each independently is selected from H, halogen, C 1-6 alkyl or deuterated alkyl or halogenated alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, 3- to 8-membered cycloalkyl or heterocycloalkyl, 5- to 10-membered aryl or heteroaryl.

[0012] Preferably, the compound of formula I is selected from the following structures:

[0013] wherein:

[0014] m, p, q, y are each independently selected from 1, 2 or 3;

[0015] R 3a , R 3b each independently is selected from R 3a , R 3b each independently is selected from H, -OH, halogen, -NH2, -NHMe, -NMe2, -CH2OH, -CH2NH2, -CH2NHMe, -CH2NMe2, -COOH, -CONH2, -CH2NHCOCHMe2, -CH2NHCOOCMe3, -CHMeNH2, -CH2COOEt, -CH2NH2COOMe, -CH2NHCOOCHMe2, -CH2NHCOMe, -CH2NHPyr, -CH2NHCONH2, -CH2NHCONHEt, -CH2NHCOCF3, -CH2NHCH2CF3, -CH2NH(Pr c ), -CH2NHCH2CHF2, -CH2NHCOCHF2, -CMe2NH2, -CONHMe, -CMe2NHMe, -CMe2NHCOCF3, CHMeNHCOCF3, R 3a and R 3b are not simultaneously H;

[0016] X is selected from -O-, -S-, -NH-, -N(Me)-, -N(Et)-, -N(Pr i ​), -N(Pr c -N(CONH2)-, -C(=NOH)- or -C(=NOH)-.

[0017] Preferably, the Selected from

[0018] Preferably, the compound of Formula I is selected from any one of the following compounds:

[0019] The salt of the compound of formula I is a salt of at least one of the following acids: galactosic acid, D-glucuronic acid, glycerophosphate, hippuric acid, hydroxyethanesulfonic acid, lactobionic acid, maleic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, neopentanoic acid, terephthalic acid, thiocyanate, cholic acid, n-dodecyl sulfate, benzenesulfonic acid, citric acid, D-glucose, glycolic acid, lactic acid, malic acid, malonic acid, mandelic acid, phosphoric acid, propionic acid, hydrochloric acid, sulfuric acid, tartaric acid, succinic acid, formic acid, hydroiodine. Acids, hydrobromic acid, methanesulfonic acid, nicotinic acid, nitric acid, orotic acid, oxalic acid, picric acid, L-pyroglutamic acid, saccharinic acid, salicylic acid, gentian acid, p-toluenesulfonic acid, valeric acid, palmitic acid, sebacic acid, stearic acid, lauric acid, acetic acid, adipic acid, carbonic acid, benzenesulfonic acid, ethanedisulfonic acid, ethylsuccinic acid, fumaric acid, 3-hydroxynaphthalene-2-carboxylic acid, 1-hydroxynaphthalene-2-carboxylic acid, oleic acid, undecenoic acid, ascorbic acid, camphoric acid, camphorsulfonic acid, dichloroacetic acid, or ethanesulfonic acid.

[0020] The compounds described in this invention, or their pharmaceutically acceptable salts, esters, stereoisomers, deuterated derivatives, or solvates, can be used to prepare USP7 inhibitors for the prevention or treatment of USP7-mediated diseases.

[0021] The diseases mediated by USP7 are any one of tumors, viral infections, inflammatory diseases, or autoimmune diseases.

[0022] The tumor diseases are bone cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, myeloproliferative disease, multiple myeloma, myelodysplastic syndrome, Hodgkin's lymphoma, non-Hodgkin's lymphoma, angioma, granuloma, xanthoma, meningeal sarcoma, glioma, neuroblastoma, astrocytoma, medulloblastoma, ependymoma, germ cell tumor (pinealoma), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, fibroma, sarcoma, esophageal cancer, gastric cancer, pancreatic cancer, large intestinal cancer, colon cancer, rectal cancer, kidney cancer, prostate cancer, lymphatic cancer, testicular cancer, interstitial cell cancer, lung cancer, liver cancer, skin cancer, basal cell carcinoma or soft tissue tumor (such as synovial sarcoma, rhabdomyosarcoma, liposarcoma, Ewing's sarcoma); the viral infectious diseases are infectious diseases caused by RNA viruses (such as influenza virus, new coronavirus), DNA viruses (such as hepatitis B virus, herpes virus) and the like.

[0023] The compound of the present application can be used for preparing a pharmaceutical composition for preventing or treating a USP7-mediated disease, which can be a capsule, a powder, a tablet, a granule, a pill, an injection, a syrup, an oral liquid, an inhalant, an ointment, a suppository or a patch, and the carrier contained in the pharmaceutical composition includes excipients, binders, disintegrants, lubricants, flavoring agents, flavoring agents, coloring agents and sweetening agents.

[0024] The compound of the present application can be used in combination with one or more other types of drugs for preventing or treating the above-mentioned diseases, including but not limited to the following combinations:

[0025] The other types of prophylactic or therapeutic drugs that can be used in combination with the compounds of the present application can be one or more anticancer drugs, including alkylating agents (such as cisplatin, cyclophosphamide, ifosfamide, melphalan, chlorambucil, bendamustine, estramustine, triaziquone, imidazole carboxamide, busulfan, dibromomannitol, cyclohexyl nitrosurea, carmustine, pyrimidine nitrosourea, methylcyclohexyl nitrosurea, methylhydrazine, and the like), antimetabolites (such as fluorouracil, cytarabine, furofurouracil, bifurouracil, mercaptopurine, sulfmercaprine sodium, thiazole purine, thioguanine, methotrexate, aminopterin, and the like), antitumor antibiotics (such as mitomycin C, bleomycin, dactinomycin, mithramycin, daunorubicin, doxorubicin, chromomycin A3, enomycin, neocarzinostatin, duocarmycin, and the like), natural anticancer drugs (such as vincristine, colchicine, camptothecin, topotecan, ellipticine, indirubin, and the like), hormone drugs (such as prednisone, hydrocortisone, hydrocortisone, dexamethasone, diethylstilbestrol, bromoacetyl hexaethylstilbestrol, propionic testosterone, methyltestosterone, nandrolone phenylpropionate, naphthoxidine, tamoxifen, and the like), immunotherapeutic agents (such as PD-1 inhibitors nivolumab and pembrolizumab, and the like; PD-L1 inhibitors atezolizumab, durvalumab, and avelumab, and the like; CTLA-4 inhibitors Ipilimumab, and the like; other immune checkpoint inhibitors; cell therapy agents, and the like), antibody drug conjugates (such as Kadcyla, and the like), kinase inhibitors (such as SHP-2 inhibitors, B-RAF inhibitors, MEK inhibitors, and BTK inhibitors, and the like), IDO inhibitors (such as Epacadostat), and the like.

[0026] Advantages: Compared with the prior art, the present application has the following advantages:

[0027] 1. The benzamide compound or its pharmaceutically acceptable salt, ester, stereoisomer, deuterated compound or solvate provided by the present application has strong inhibitory activity on USP7; and can be used for preparing a drug for preventing or treating a USP7-mediated disease.

[0028] 2. The benzamide compound or its pharmaceutically acceptable salt, ester, stereoisomer, deuterated compound or solvate provided by the present application has low hERG (human ether-a-go-go) potassium ion channel inhibitory effect, indicating that the risk of cardiotoxic side effects is small.

[0029] 3. The benzamide compound or its pharmaceutically acceptable salt, ester, stereoisomer, deuterated compound or solvate provided by the present application has high metabolic stability in human liver microsomes, indicating that the PD / PK property is good, and the drug potential is high. DETAILED DESCRIPTION

[0030] The application will be further described in conjunction with specific examples. The materials, reagents and the like used in the examples, unless otherwise specified, can be obtained from commercial sources. The experimental methods not specified in the examples are usually carried out according to the conventional conditions or the conditions recommended by the manufacturers.

[0031] Example 1

[0032] Preparation of 3-((7-(5-chloro-2-(1,4-diazepane-1-carbonyl)-3-methylphenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 1).

[0033] (1) 7-chlorothieno[3,2-b]pyridine (2 g, 12 mmol) was dissolved in anhydrous tetrahydrofuran (100 mL), replaced with argon for three times, cooled to -78 °C, and then slowly dropped with n-butyllithium in n-hexane (2.5 M, 6.13 mL, 15 mmol) by using a syringe pump. After dropping, it was stirred at -78 °C for 1 hour, and then anhydrous N,N-dimethylformamide (3.05 mL, 40 mmol) was slowly dropped. After dropping, it was reacted for 1 hour, and then the reaction was completed by monitoring with TLC (petroleum ether: ethyl acetate = 5:1). The reaction was quenched by dropping saturated ammonium chloride solution into the system, and then the solvent was removed under reduced pressure. Water (150 mL) was added to the residue to make a slurry, and then it was filtered under suction. The filter cake was washed with a small amount of water, and then it was dried to obtain the intermediate A1 as a yellowish brown solid (2.37 g, crude yield 100%). The crude product was directly used in the next reaction without purification. 1 H NMR (300 MHz, Chloroform-d) δ 10.21 (s, 1H), 8.72 (d, J = 5.0 Hz, 1H), 8.25 (s, 1H), 7.45 (d, J = 5.0 Hz, 1H).

[0034] (2) The intermediate A1 (1.71 g, 8.70 mmol) was placed in methanol (10 mL), and then sodium borohydride (0.64 g, 17 mmol) was added in batches under ice bath. Then it was slowly warmed to room temperature and reacted for 1 hour. After the reaction was completed by monitoring with TLC (petroleum ether: ethyl acetate = 2:1), it was quenched by adding saturated ammonium chloride solution. The methanol was removed under reduced pressure, and then water (100 mL) was added to the residue to make a slurry. It was filtered under suction, and then the filter cake was washed with a small amount of water. After drying, the intermediate A2 was obtained as a yellow solid (1.73 g, crude yield 100%). The crude product was directly used in the next reaction without purification. 1H NMR (400 MHz, Chloroform-d) δ 8.54 (d, J = 5.1 Hz, 1H), 7.44 (t, J = 1.1 Hz, 1H), 7.26 (d, J = 5.2 Hz, 1H), 5.01 (d, J = 1.1 Hz, 2H).

[0035] (3) Intermediate A2 (1.1 g, 5.51 mmol) was placed in anhydrous dichloromethane (30 mL), and oxysulfur chloride (1.2 mL, 16.53 mmol) was slowly added dropwise under ice bath, and stirred at room temperature overnight after dropwise addition. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 2:1), and the reaction was completed. Saturated sodium bicarbonate solution was slowly added dropwise under ice bath to quench, extracted with dichloromethane (80 mL x 2), the combined organic phase was washed with saturated sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 15:1) to obtain white solid of intermediate A3 (1.60 g, yield 86%). 1 H NMR (400 MHz, Chloroform-d) δ 8.54 (d, J = 5.1 Hz, 1H), 7.44 (t, J = 1.1 Hz, 1H), 7.26 (d, J = 5.2 Hz, 1H), 5.01 (d, J = 1.1 Hz, 2H).

[0036] (4) Intermediate A3 (3.9 g, 18 mmol) and 6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (3.8 g, 27 mmol) were dissolved in acetonitrile (60 mL), and potassium carbonate (5 g, 36 mmol) was added. The reaction was heated to 80 °C for 10 hours. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 1:1), and the reaction was completed. The reaction was cooled to room temperature, and water (150 mL) was added. The organic phase was extracted with dichloromethane (80 mL x 3), and the combined organic phase was washed with saturated sodium chloride aqueous solution (100 mL), dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain pink solid of intermediate A4 (5.52 g, yield 96%). 1 H NMR (400 MHz, Chloroform-d) δ 8.54 (d, J = 5.1 Hz, 1H), 7.44 (t, J = 1.1 Hz, 1H), 7.26 (d, J = 5.2 Hz, 1H), 5.01 (d, J = 1.1 Hz, 2H).

[0037] (5) Intermediate A4 (0.5 g, 1.56 mmol), bis(pinacolato)diboron (0.43 g, 1.71 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (0.063 g, 0.085 mmol) and potassium acetate (0.31 g, 3.12 mmol) were added to anhydrous dimethyl sulfoxide (8 mL), replaced with argon for three times, and warmed to 100 °C for 16 hours. TLC (petroleum ether: ethyl acetate = 1:1) was used to monitor the completion of the reaction. The reaction solution was cooled to room temperature, diluted with ethyl acetate (20 mL), and filtered with diatomite. Water (20 mL) was added to the filtrate, and extracted with ethyl acetate (20 mL x 3). The combined organic phase was washed with water (20 mL x 2), saturated sodium chloride aqueous solution (20 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was slurried with n-hexane (15 mL), filtered, and dried to obtain intermediate A5 as a dark brown solid (0.79 g, crude yield 100%), which was used directly in the next step without purification.

[0038] (6) 4-Chloro-2-methylbenzoic acid (15 g, 88 mmol), diacetoxyiodobenzene (56 g, 170 mmol), iodine (43 g, 170 mmol), palladium acetate (0.99 g, 4.4 mmol) were added to anhydrous N,N-dimethylformamide (200 mL), replaced with argon for three times, and warmed to 80 °C for 8 hours. TLC (petroleum ether: ethyl acetate = 1:1, plus 1 drop of acetic acid) was used to monitor the completion of the reaction. The reaction solution was cooled to room temperature, diluted with methyl tert-butyl ether (20 mL), and then quenched with sodium pyrosulfite. The pH was adjusted to 1 by adding concentrated hydrochloric acid, and extracted with methyl tert-butyl ether (200 mL x 3). The combined organic phase was adjusted to pH 14 with saturated sodium hydroxide aqueous solution at 0 °C, and the organic phase was discarded. The aqueous phase was adjusted to pH 1 with concentrated hydrochloric acid at 0 °C, and extracted with methyl tert-butyl ether (200 mL x 3). The solvent was removed under reduced pressure to obtain intermediate A6 as a yellow solid (20 g, yield 76%). 1 H NMR (300 MHz, Chloroform-d) δ 8.09 (s, 1H), 7.69 (dd, J = 1.9, 0.7 Hz, 1H), 7.21 (dd, J = 1.9, 0.7 Hz, 1H), 2.41 (s, 3H).

[0039] (7) Intermediate A6 (7 g, 24 mmol) was dissolved in thionyl chloride (20 mL) and heated at 80 °C for 3 h. After A6 was completely converted to acyl chloride, the solvent was removed under reduced pressure. tert-Butyl 1,4-diazepane-1-carboxylate (4.72 g, 24 mmol) was dissolved in anhydrous dichloromethane (72 mL), and triethylamine (6.54 mL, 48 mmol) was added. The acyl chloride solution in anhydrous dichloromethane (20 mL) was added dropwise at 0 °C, and the mixture was allowed to warm to room temperature for 0.5 h. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 1:1), and the reaction was quenched by the addition of saturated sodium bicarbonate solution. The mixture was extracted with dichloromethane (50 mL x 3), and the combined organic phases were washed with saturated aqueous sodium chloride solution (30 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 5:1) to give intermediate A7 as a yellow-white foamy solid (11 g, 96%). + .

[0040] (8) A7 (1.05 g, 2.2 mmol), A5 (1.35 g, 3.3 mol), Pd(amphos)2Cl2(0.078 g, 0.11 mmol), and potassium carbonate (0.61 g, 4.4 mmol) were added to a mixture of dioxane / water (12 mL / 3 mL), and the mixture was replaced with argon three times. The mixture was warmed to 100 °C and stirred for 8 h. The reaction was monitored by TLC (dichloromethane:methanol = 30:1), and the mixture was filtered through celite. The filtrate was extracted with ethyl acetate (40 mL x 3), and the combined organic phases were washed with saturated aqueous sodium chloride solution (20 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 1:1) to give intermediate A8 as a yellow solid (0.97 g).

[0041] (9) Intermediate A8 (970 mg, 1.59 mmol) was dissolved in ethyl acetate (8 mL), and hydrogen chloride in ethyl acetate (4 N, 3 mL) was added at 0 °C. The mixture was stirred at room temperature for 3 h. The reaction was monitored by TLC (dichloromethane:methanol = 15:1), and the reaction was quenched by the addition of saturated aqueous sodium bicarbonate solution (30 mL). The mixture was extracted with ethyl acetate (40 mL x 3), and the combined organic phases were washed with saturated aqueous sodium chloride solution (20 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative TLC (dichloromethane:methanol = 10:1) to give the target compound 1 as a white solid (94 mg, 10% over two steps). 1H NMR (500 MHz, DMSO-d6, Amide isomerization of two sets of peaks, 80 °C high temperature nuclear magnetic can make two sets of peaks degenerate) δ 8.81 (s, 1H), 8.68 (d, J = 9.4, 4.8 Hz, 1H), 7.57 (d, J = 2.2 Hz, 1H), 7.51 (d, J = 2.2 Hz, 2H), 7.24 (d, J = 4.8 Hz, 1H), 4.80 (s, 2H), 3.59 (t, J = 4.8 Hz, 2H), 3.26 - 3.10 (m, 3H), 2.89 - 2.74 (m, 2H), 2.56 (s, 1H), 2.32 (s, 2H), 2.13 - 2.06 (m, 1H), 1.87 - 1.68 (m, 1H), 1.53 - 1.37 (m, 1H), 1.21 (s, 3H), 1.07 (s, 3H). HRMS (ESI): m / z exact mass calculated for C 28 H 30 ClN4O3S[M+H] + 537.1727, found 537.1734.

[0042] Example 2

[0043] 3-((7-(5-chloro-2-(1,5-diazepan-1- carbonyl)-3-methylphenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 2) was prepared according to the procedure described in Reference Example 1, except that tert-butyl 1,4-diazepane-1- carboxylate was replaced by tert-butyl 1,5-diazepane-1-carboxylate to give Compound 2 as a white solid (57 mg, 3% yield over two steps).

[0044] 3-((7-(5-chloro-2-(1,5-diazepan-1- carbonyl)-3-methylphenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 2) was prepared according to the procedure described in Reference Example 1, except that tert-butyl 1,4-diazepane-1- carboxylate was replaced by tert-butyl 1,5-diazepane-1-carboxylate to give Compound 2 as a white solid (57 mg, 3% yield over two steps). 1 H NMR (300 MHz, DMSO-d6) δ 8.69 (d, J = 4.8 Hz, 1H), 8.41 (s, 1H), 7.66 - 7.54 (m, 3H), 7.25 (d, J = 4.7 Hz, 1H), 4.80 (s, 2H), 3.55 - 3.39 (m, 2H), 3.14 - 2.96 (m, 2H), 2.95 - 2.80 (m, 2H), 2.80 - 2.64 (m, 2H), 2.61 (s, 2H), 2.31 (s, 3H), 2.09 - 1.93 (m, 1H), 1.88 - 1.70 (m, 2H), 1.65 - 1.49 (m, 1H), 1.17 (s, 3H), 1.03 (s, 3H). HRMS (ESI): m / z exact mass calculated for C29 H 32 ClN4O3S[M+H] + 551.1884, found 551.1885.

[0045] Example 3

[0046] Preparation of 3-((7-(5-chloro-3-methyl-2-(5-oxo-1,4-azepane-1-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 3).

[0047] Refer to Example 1, the only difference is that 1,4-diazepane-1-carboxylic acid tert-butyl ester is replaced by 1,4-diazepan-5-one, no need to remove Boc after coupling, white solid of Compound 3 is prepared (58 mg, 11%). 1 H NMR (300 MHz, DMSO-d6) d 8.69 (d, J = 4.8 Hz, 1H), 7.69 - 7.62 (m, 1H), 7.64 - 7.54 (m, 2H), 7.54 (d, J = 7.2 Hz, 1H), 7.23 (dd, J = 4.9, 2.9 Hz, 1H), 4.80 (s, 2H), 3.66 - 3.46 (m, 1H), 3.47 - 3.34 (m, 2H), 3.22 - 2.95 (m, 2H), 2.93 - 2.75 (m, 1H), 2.60 (d, J = 3.3 Hz, 2H), 2.27 (s, 3H), 2.21 - 2.05 (m, 1H), 1.89 (dd, J = 15.2, 8.7 Hz, 1H), 1.17 (s, 3H), 1.02 (s, 3H). HRMS (ESI): m / z exact mass calculated for C 28 H 28 ClN4O4S[M+H] + 551.1520, found 551.1523.

[0048] Example 4

[0049] Preparation of 3-((7-(5-chloro-3-methyl-2-(3-oxo-1,4-azepane-1-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 4).

[0050] Reference to Example 1, except that tert-butyl 1,4-diazepane-1-carboxylate was replaced by 1,4-diazepan-3-one, and no Boc removal was needed after coupling, to give compound 4 as a white solid (3 mg, 1%). 1 H NMR (300 MHz, DMSO-d6) δ 8.69 (d, J = 4.8 Hz, 1H), 7.71 - 7.60 (m, 2H), 7.59 - 7.56 (m, 1H), 7.56 - 7.48 (m, 1H), 7.28 - 7.01 (m, 1H), 4.80 (s, 2H), 3.72 - 3.40 (m, 2H), 3.27 - 2.96 (m, 2H), 2.96 - 2.66 (m, 2H), 2.60 (d, J = 3.3 Hz, 2H), 2.43 - 2.32 (m, 1H), 2.27 (s, 3H), 1.99 - 1.86 (m, 1H), 1.17 (d, J = 2.7 Hz, 3H), 1.02 (d, J = 1.8 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C 28 H 28 ClN4O4S[M+H] + 551.1520, found 551.1523.

[0051] Example 5

[0052] 3-((7-(5-chloro-3-methyl-2-(1,4-oxazepane-4-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 5).

[0053] Reference to Example 1, except that tert-butyl 1,4-diazepane-1-carboxylate was replaced by 1,4-diazepan-3-one, and no Boc removal was needed after coupling, to give compound 5 as a white solid (55 mg, 6%). 1H NMR (300 MHz, DMSO-d6) δ 8.68 (d, J = 6.7, 4.8 Hz, 1H), 7.64 - 7.53 (m, 2H), 7.55 (d, J = 3.4 Hz, 1H), 7.32 - 7.24 (m, 1H), 4.79 (s, 2H), 3.63 - 3.36 (m, 3H), 3.30 - 3.14 (m, 1H), 3.13 - 2.91 (m, 2H), 2.89 - 2.70 (m, 2H), 2.60 (s, 2H), 2.28 (s, 3H), 1.70 - 1.27 (m, 2H), 1.16 (s, 3H), 1.01 (d, J = 2.9 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C 28 H 29 ClN3O4S[M+H] + 538.1567, found 538.1567.

[0054] Example 6

[0055] Preparation of 3-((7-(5-chloro-3-methyl-2-(2-oxa-6-azaspiro[3.3]heptane-6- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane- 2,4-dione (Compound 6).

[0056] Refer to Example 1, the only difference is that 1,4-diazepane-1-carboxylic acid tert-butyl ester is replaced by 2-oxa-6-azaspiro[3,3]heptane, no need to remove Boc after coupling, white solid of Compound 6 is prepared (43 mg, 26%). 1 H NMR (300 MHz, DMSO-d6) δ 8.68 (d, J = 6.7, 4.8 Hz, 1H), 7.64 - 7.53 (m, 2H), 7.55 (d, J = 3.4 Hz, 1H), 7.32 - 7.24 (m, 1H), 4.79 (s, 2H), 3.63 - 3.36 (m, 3H), 3.30 - 3.14 (m, 1H), 3.13 - 2.91 (m, 2H), 2.89 - 2.70 (m, 2H), 2.60 (s, 2H), 2.28 (s, 3H), 1.70 - 1.27 (m, 2H), 1.16 (s, 3H), 1.01 (d, J = 2.9 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C28 H 27 ClN3O4S[M+H] + 536.1411, found 536.1411.

[0057] Example 7

[0058] Preparation of 3-((7-(5-chloro-3-methyl-2-(2,7-diazaspiro[3.5]nonane-7- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 7).

[0059] Referring to Example 1, except that tert-butyl 2,7-diazaspiro[3.5]nonane-2- carboxylate was used instead of tert-butyl 1,4-diazepane-1-carboxylate, Compound 7 was prepared as a white solid (71 mg, 8% yield over two steps). 1 H NMR (300 MHz, DMSO-d6) δ 9.23 (s, 1H), 8.65 (d, J = 4.8 Hz, 1H), 7.59 (d, J = 2.1 Hz, 1H), 7.54 (d, J = 2.7 Hz, 2H), 7.18 (d, J = 4.8 Hz, 1H), 4.79 (s, 2H), 3.59 (s, 2H), 3.53 - 3.42 (m, 1H), 3.38 - 3.17 (m, 3H), 3.05 - 2.86 (m, 1H), 2.77 - 2.59 (m, 3H), 2.25 (s, 3H), 1.74 - 1.61 (m, 1H), 1.49 - 1.34 (m, 1H), 1.19 (s, 3H), 1.13 - 1.08 (m, 1H), 1.06 (s, 3H), 0.76 - 0.59 (m, 1H). HRMS (ESI): m / z exact mass calculated for C 30 H 32 ClN4O3S[M+H] + 563.1884, found 563.1885.

[0060] Example 8

[0061] Preparation of 3-((7-(5-chloro-3-methyl-2-(2,8-diazaspiro[4.5]decane-8- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 8).

[0062] Reference to Example 1, except that tert-butyl 2,8-diazaspiro[4.5]decane-2- carboxylate was used instead of tert-butyl 1,4-diazepane-1-carboxylate, to afford Compound 8 as a white solid (76 mg, 19% yield over two steps). 1 H NMR (300 MHz, DMSO-d6) δ 9.22 (s, 1H), 8.68 (d, J = 4.8, 1.9 Hz, 1H), 7.62 - 7.51 (m, 3H), 7.26 - 7.18 (m, 1H), 4.79 (s, 2H), 3.66 - 3.43 (m, 1H), 3.17 - 2.92 (m, 4H), 2.92 - 2.70 (m, 2H), 2.70 - 2.58 (m, 3H), 2.27 (s, 3H), 1.84 - 1.56 (m, 1H), 1.48 - 1.35 (m, 2H), 1.18 (s, 2H), 1.15 - 1.07 (m, 2H), 1.06 (s, 2H), 0.78 - 0.70 (m, 1H). HRMS (ESI): m / z exact mass calculated for C 31 H 34 ClN4O3S[M+H] + 577.2040, found 577.2040.

[0063] Example 9

[0064] 3-((7-(5-chloro-3-methyl-2-(3,9-diazaspiro[5.5]undecane-3-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 9).

[0065] Reference to Example 1, except that tert-butyl 2,8-diazaspiro[4.5]decane-2- carboxylate was used instead of tert-butyl 1,4-diazepane-1-carboxylate, to afford Compound 8 as a white solid (76 mg, 19% yield over two steps). 1H NMR (300 MHz, DMSO-d6) δ 8.68 (d, J = 4.8 Hz, 2H), 7.60 (d, J = 0.7 Hz, 2H), 7.54 (s, 1H), 7.23 (d, J = 4.8 Hz, 1H), 4.88 - 4.70 (m, 2H), 3.78 - 3.50 (m, 1H), 3.14 - 2.70 (m, 7H), 2.71 - 2.59 (m, 2H), 2.26 (s, 3H), 1.72 - 1.29 (m, 4H), 1.19 (s, 3H), 1.05 (s, 3H), 1.02 - 0.75 (m, 4H). HRMS (ESI): m / z exact mass calculated for C 32 H 36 ClN4O3S [M+H] + 591.2197, found 591.2201.

[0066] Example 10

[0067] 3-((7-(5-chloro-3-methyl-2-(2,7-diazaspiro[4.4]nonane-2-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 10) was prepared according to the procedure described in Reference Example 1, except that tert-butyl 1,4-diazepane-1-carboxylate was replaced by tert-butyl 2,7-diaza-spiro[4.4]nonane-2-carboxylate, to give Compound 10 as a white solid (96 mg, 3% yield over two steps).

[0068] 3-((7-(5-chloro-3-methyl-2-(2,7-diazaspiro[4.4]nonane-2-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 10) was prepared according to the procedure described in Reference Example 1, except that tert-butyl 1,4-diazepane-1-carboxylate was replaced by tert-butyl 2,7-diaza-spiro[4.4]nonane-2-carboxylate, to give Compound 10 as a white solid (96 mg, 3% yield over two steps). 1 H NMR (400 MHz, DMSO-d6) δ 9.19 (s, 1H), 8.72 - 8.65 (m, 1H), 7.67 - 7.51 (m, 3H), 7.31 - 7.20 (m, 1H), 4.80 (s, 2H), 3.41 - 3.24 (m, 3H), 3.19 - 3.07 (m, 1H), 3.07 - 2.96 (m, 2H), 2.96 - 2.85 (m, 1H), 2.74 - 2.68 (m, 1H), 2.66 - 2.57 (m, 2H), 2.34 - 2.28 (m, 3H), 1.86 - 1.70 (m, 2H), 1.58 - 1.40 (m, 1H), 1.35 (s, 1H), 1.24 - 1.14 (m, 3H), 1.13 - 0.92 (m, 3H). HRMS (ESI): m / z exact mass calculated for C 30 H 32 ClN4O3S [M+H]+ 563.1884, found 563.1885.

[0069] Example 11

[0070] Preparation of 3-((7-(5-chloro-3-methyl-2-(2,7-diazaspiro[3.5]decane-2- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane- 2,4-dione (Compound 11).

[0071] Referring to Example 1, except for replacing 1,4-diazepane-1-carboxylic acid tert-butyl ester with 2,7-diazaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester, Compound 11 was prepared as a white solid (61 mg, 13% yield over two steps). 1 H NMR (400 MHz, DMSO-d6) d 8.71 (d, J = 4.8 Hz, 1H), 7.60 (d, J = 2.1 Hz, 1H), 7.58 (s, 1H), 7.57 (s, 1H), 7.23 (d, J = 4.8 Hz, 1H), 4.80 (s, 2H), 3.62 - 3.55 (m, 1H), 3.51 - 3.35 (m, 2H), 3.17 - 3.10 (m, 1H), 2.99 - 2.84 (m, 1H), 2.79 - 2.69 (m, 1H), 2.66 - 2.59 (m, 3H), 2.59 - 2.52 (m, 1H), 2.33 (s, 3H), 1.88 (s, 1H), 1.58 (t, J = 5.6 Hz, 2H), 1.18 (s, 3H), 1.03 (s, 3H), 0.98 - 0.88 (m, 1H), 0.66 - 0.58 (m, 1H). HRMS (ESI): m / z exact mass calculated for C 30 H 32 ClN4O3S [M+H] + 563.1884, found 563.1885.

[0072] Example 12

[0073] Preparation of 3-((7-(5-chloro-3-methyl-2-(2,6-diazaspiro[3.3]heptane-2- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane- 2,4-dione (Compound 12).

[0074] Referring to Example 1, except that tert-butyl 2,6-diazaspiro[3.3]heptane-2- carboxylate was used instead of tert-butyl 1,4-diazepane-1-carboxylate, Compound 12 was prepared as a white solid (134 mg, 15% yield over two steps). 1 H NMR (400 MHz, DMSO-d6) δ 8.69 (d, J = 4.8 Hz, 1H), 7.62 - 7.58 (m, 1H), 7.58 - 7.51 (m, 2H), 7.30 - 7.17 (m, 1H), 4.79 (d, J = 3.6 Hz, 2H), 3.76 (s, 1H), 3.73 - 3.56 (m, 1H), 3.48 - 3.38 (m, 2H), 3.22 - 3.13 (m, 1H), 3.09 - 3.01 (m, 1H), 3.00 - 2.92 (m, 1H), 2.67 - 2.61 (m, 1H), 2.60 - 2.56 (m, 2H), 2.35 (s, 3H), 1.16 (s, 3H), 1.02 (d, J = 2.9 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C 28 H 28 ClN4O3S[M+H] + 535.1571, found 535.1572.

[0075] Example 13

[0076] Preparation of 3-((7-(5-chloro-3-methyl-2-(hexahydropyrrolo[3,4-c]pyrrole-2- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane- 2,4-dione (Compound 13).

[0077] Referring to Example 1, except that tert-butyl 2,6-diazaspiro[3.3]heptane-2- carboxylate was used instead of tert-butyl 1,4-diazepane-1-carboxylate, Compound 12 was prepared as a white solid (134 mg, 15% yield over two steps). 1H NMR (400 MHz, DMSO-d6) δ 8.92 (s, 1H), 8.75 - 8.64 (m, 1H), 7.68 - 7.59 (m, 1H), 7.58 - 7.55 (m, 1H), 7.54 (s, 1H), 7.31 - 7.14 (m, 1H), 4.80 (s, 2H), 3.69 - 3.34 (m, 3H), 3.28 - 3.09 (m, 3H), 3.04 - 2.91 (m, 1H), 2.91 - 2.83 (m, 1H), 2.82 - 2.72 (m, 2H), 2.63 - 2.57 (m, 2H), 2.36 - 2.26 (m, 3H), 1.17 (s, 3H), 1.05 - 0.99 (m, 3H). HRMS (ESI): m / z exact mass calculated for C 29 H 30 ClN4O3S[M+H] + 549.1727, found 549.1730.

[0078] Example 14

[0079] Preparation of 4-(1,4-diazepane-1 -carbonyl)-3-(2-((6,6-dimethyl-2,4-dioxo-3- azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2-b]pyridin-7-yl)-5-methylbenzonitrile (Compound 14).

[0080] Referring to the preparation method of compounds A6, A7, A8 and 1 in Example 1, the only difference is that 4-chloro-2-methylbenzoic acid is replaced by 4-cyano-2-methylbenzoic acid, white solid of compound 14 is prepared (10 mg, yield 4%). 1HNMR (300 MHz, DMSO-d6) δ 9.11 (s, 1H), 8.71 (dd, J = 7.8, 4.8 Hz, 1H), 8.06 - 7.99 (m, 2H), 7.56 (d, J = 3.5 Hz, 1H), 7.25 (t, J = 4.4 Hz, 1H), 4.80 (s, 2H), 4.40 (t, J = 5.0 Hz, 1H), 3.57 (s, 2H), 3.43 (dd, J = 7.0, 4.9 Hz, 1H), 3.25 - 3.03 (m, 2H), 2.76 - 2.70 (m, 2H), 2.62 (d, J = 1.3 Hz, 2H), 2.34 (d, J = 1.7 Hz, 3H), 1.90 - 1.77 (m, 1H), 1.48 (s, 1H), 1.17 (s, 3H), 1.05 (s, 3H). HRMS (ESI): m / z exact mass calculated for C 29 H 30 N5O3S[M+H] + 528.2069, found 528.2077.

[0081] Example 15

[0082] 3-((7-(5-chloro-3-methyl-2-(1-oxa-4,9-diazaspiro[5.5]undecane-9-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 15).

[0083] Following Example 1, except replacing 1,4-diazepane-1-carboxylic acid tert-butyl ester with 1-oxa-4,9-diazaspiro[5.5]undecane-4-carboxylic acid tert-butyl ester, Compound 15 was prepared as a white solid (25 mg, yield 6%). 1H NMR (300 MHz, DMSO-d6) δ 8.93 (s, 1H), 8.68 (dd, J = 11.2, 4.8 Hz, 1H), 7.62 - 7.57 (m, 1H), 7.54 (s, 1H), 7.47 - 7.25 (m, 1H), 7.24 - 7.14 (m, 1H), 4.80 (d, J = 3.0 Hz, 2H), 3.98 - 3.81 (m, 1H), 3.71 - 3.49 (m, 2H), 3.12 - 2.67 (m, 5H), 2.62 (d, J = 6.9 Hz, 2H), 2.48 - 2.33 (m, 2H), 2.32 - 2.22 (m, 3H), 2.02 - 1.63 (m, 1H), 1.40 - 1.21 (m, 3H), 1.18 (d, J = 4.5 Hz, 3H), 1.08 - 0.97 (m, 3H). HRMS (ESI): m / z exact mass calculated for C 31 H 34 ClN4O4S[M+H] + 593.1911, found 593.1974.

[0084] Example 16

[0085] Preparation of 3-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2- b]pyridin-7-yl)-5-methyl-4-(1-oxa-4,9-diazaspiro[5.5]undecane-9-carbonyl)benzonitrile (Compound 16).

[0086] Referring to the preparation method of compounds A6, A7, A8 and 1 in Example 1, the only difference is that the intermediate 4-chloro-2-methylbenzoic acid is replaced by 4-cyano-2-methylbenzoic acid, and the intermediate tert-butyl 1,4-diazepane-1-carboxylate is replaced by tert-butyl 1-oxa-4,9-diazaspiro[5.5]undecane-4-carboxylate, to prepare white solid of compound 16 (45 mg, yield 7.4%). 1H NMR (400 MHz, DMSO-d6) δ 8.74 - 8.65 (m, 1H), 8.10 - 7.96 (m, 2H), 7.58 - 7.52 (m, 1H), 7.27 - 7.16 (m, 1H), 3.99 - 3.79 (m, 1H), 3.65 - 3.42 (m, 2H), 3.07 - 2.75 (m, 2H), 2.72 - 2.64 (m, 2H), 2.61 (d, J = 5.7 Hz, 2H), 2.36 - 2.26 (m, 3H), 2.14 (s, 1H), 1.96 - 1.59 (m, 2H), 1.41 - 1.23 (m, 2H), 1.17 (d, J = 4.8 Hz, 3H), 1.08 - 0.99 (m, 3H). HRMS (ESI): m / z exact mass calculated for C 32 H 34 N5O4S[M+H] + 584.2253, found 584.2316.

[0087] Example 17

[0088] 3-((7-(2-(4-(Aminomethyl)piperidine-l-carbonyl)-5-chloro-3-methylphenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 17) was prepared according to the procedure described in Reference Example 1, except that tert-butyl 1,4-diazepane-l-carboxylate was replaced by tert-butyl (piperidin-4- ylmethyl)carbamate, to give Compound 17 as a white solid (5.6 mg, yield 1.5%).

[0089] 3-((7-(2-(4-(Aminomethyl)piperidine-l-carbonyl)-5-chloro-3-methylphenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 17) was prepared according to the procedure described in Reference Example 1, except that tert-butyl 1,4-diazepane-l-carboxylate was replaced by tert-butyl (piperidin-4- ylmethyl)carbamate, to give Compound 17 as a white solid (5.6 mg, yield 1.5%). 1 H NMR (400 MHz, DMSO-d6) δ 8.69 - 8.65 (m, 1H), 7.62 - 7.49 (m, 3H), 7.27 - 7.12 (m, 1H), 5.98 - 5.30 (m, 1H), 4.79 (s, 2H), 4.31 - 4.19 (m, 1H), 3.20 - 3.10 (m, 1H), 3.04 - 2.96 (m, 1H), 2.87 - 2.74 (m, 1H), 2.62 - 2.57 (m, 2H), 2.35 - 2.14 (m, 5H), 2.14 - 1.87 (m, 1H), 1.62 - 1.51 (m, 1H), 1.51 - 1.22 (m, 3H), 1.18 (d, J = 2.5 Hz, 3H), 1.08 - 1.00 (m, 3H). HRMS (ESI): m / z exact mass calculated for C 29 H31 CIN4O3S [M+H] + 551.1884, found 551.1871.

[0090] Example 18

[0091] Preparation of 3-((7-(2-(3-(aminomethyl)azetidine-1-carbonyl)-5-chloro-3- methylphenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane- 2,4-dione (Compound 18).

[0092] Referring to Example 1, the only difference is that tert-butyl 1,4-diazepane-1-carboxylate is replaced by tert-butyl carbamate (azetidin-3-ylmethyl) to give Compound 18 as a white solid (26 mg, yield 4.8%). 1 H NMR (400 MHz, DMSO-d6) δ 8.75 - 8.67 (m, 1H), 7.80 (s, 2H), 7.62 - 7.58 (m, 1H), 7.55 (d, J = 3.4 Hz, 1H), 7.55 - 7.52 (m, 1H), 7.27 - 7.20 (m, 1H), 4.80 (s, 2H), 4.12 - 3.99 (m, 1H), 3.99 - 3.81 (m, 1H), 3.73 - 3.58 (m, 1H), 3.50 - 3.42 (m, 1H), 3.17 (d, J = 5.1 Hz, 1H), 3.02 - 2.87 (m, 2H), 2.60 (s, 2H), 2.35 (d, J = 12.5 Hz, 3H), 1.17 (s, 3H), 1.02 (d, J = 6.8 Hz, 3H).

[0093] Example 19

[0094] Preparation of 1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3- yl)methyl)thieno[3,2-b]pyridin-7-yl)-6-methylbenzoyl)piperidine-4-carboxamide (Compound 19).

[0095] Referring to Example 1, the only difference is that tert-butyl 1,4-diazepane-1-carboxylate is replaced by piperidine-4-carboxamide reaction, no need to remove Boc after coupling to give Compound 19 as a white solid (76 mg, yield 12.9% for two steps). 1H NMR (400 MHz, DMSO-d6) δ 8.69 - 8.63 (m, 1H), 7.58 (dd, J = 7.3, 2.1 Hz, 1H), 7.55 - 7.49 (m, 2H), 7.26 - 6.98 (m, 2H), 6.79 - 6.60 (m, 1H), 4.84 - 4.71 (m, 2H), 4.23 - 4.10 (m, 1H), 3.23 - 3.01 (m, 1H), 2.91 - 2.63 (m, 1H), 2.60 (s, 1H), 2.59 - 2.55 (m, 1H), 2.33 - 2.23 (m, 4H), 2.15 - 2.01 (m, 1H), 1.62 - 1.50 (m, 1H), 1.41 - 1.18 (m, 3H), 1.17 (d, J = 2.8 Hz, 3H), 1.01 (d, J = 1.3 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C 29 H 29 ClN4O4S[M+H] + 564.1598, found 565.1658.

[0096] Example 20

[0097] 3-((7-(5-chloro-3-methyl-2-(piperazin-l- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 20) was prepared according to the procedure described in Example 1, with the exception that 1,4-diazepane-l-carboxylic acid tert-butyl ester was replaced by piperazine- 1-carboxylic acid tert-butyl ester to give Compound 20 as a white solid (13 mg, 15% yield).

[0098] 3-((7-(5-chloro-3-methyl-2-(piperazin-l- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 20) was prepared according to the procedure described in Example 1, with the exception that 1,4-diazepane-l-carboxylic acid tert-butyl ester was replaced by piperazine- 1-carboxylic acid tert-butyl ester to give Compound 20 as a white solid (13 mg, 15% yield). 1 H NMR (300 MHz, DMSO-d6) δ 8.69 (d, J = 2.1 Hz, 1H), 7.57 (d, J = 2.1 Hz, 1H), 7.55 (s, 1H), 7.22 (d, J = 4.8 Hz 1H), 4.80 (s, 2H), 3.60 - 3.41 (m, 6H), 2.95-2.82 (m, 2H), 2.60 (s, 2H), 1.16 (s, 3H), 1.01 (s, 3H). HRMS (ESI): m / z exact mass calculated for C 27 H 27 ClN4O4S[M+H] + 523.1571, found 523.1568.

[0099] Example 21

[0100] 3-((7-(5-chloro-3-methyl-2-(1-oxo-2,9-diazaspiro[5.5]undecan-9-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 21) was prepared according to the procedure described in Example 1, with the exception that tert-butyl 1,4-diazepane-1-carboxylate was replaced by tert-butyl 1-oxo-2,9-diazaspiro[5.5]undecan-9-carboxylate, which was first de-Boc under acidic condition before it was engaged in the reaction with A6. No de-Boc was needed after the coupling. Compound 21 was obtained as a yellow solid (160 mg, 35.5%).

[0101] 3-((7-(5-chloro-3-methyl-2-(1-oxo-2,9-diazaspiro[5.5]undecan-9-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 21) was prepared according to the procedure described in Example 1, with the exception that tert-butyl 1,4-diazepane-1-carboxylate was replaced by tert-butyl 1-oxo-2,9-diazaspiro[5.5]undecan-9-carboxylate, which was first de-Boc under acidic condition before it was engaged in the reaction with A6. No de-Boc was needed after the coupling. Compound 21 was obtained as a yellow solid (160 mg, 35.5%). 1 H NMR (400 MHz, DMSO-d6) d 8.71 - 8.62 (m, 1H), 7.61 - 7.48 (m, 3H), 7.28 - 7.14 (m, 2H), 4.82 - 4.75 (m, 2H), 3.64 - 3.53 (m, 1H), 3.32 - 3.16 (m, 3H), 3.05 - 2.84 (m, 3H), 2.60 - 2.56 (m, 2H), 2.28 (d, J = 5.1 Hz, 3H), 1.77 - 1.66 (m, 1H), 1.65 - 1.32 (m, 4H), 1.16 (d, J = 5.0 Hz, 4.5H), 1.04 - 0.96 (m, 3H), 0.79 - 0.70 (m, 0.5H). HRMS (ESI): m / z exact mass calculated for C 32 H 33 ClN4O4S[M+H] + 604.1911, found 605.1973.

[0102] Example 22

[0103] 3-((7-(5-chloro-3-methyl-2-(1-oxo-2,9-diazaspiro[5.5]undecan-9-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 21) was prepared according to the procedure described in Example 1, with the exception that tert-butyl 1,4-diazepane-1-carboxylate was replaced by tert-butyl 1-oxo-2,9-diazaspiro[5.5]undecan-9-carboxylate, which was first de-Boc under acidic condition before it was engaged in the reaction with A6. No de-Boc was needed after the coupling. Compound 21 was obtained as a yellow solid (160 mg, 35.5%).

[0104] 3-((7-(5-chloro-3-methyl-2-(1-oxo-2,9-diazaspiro[5.5]undecan-9-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 21) was prepared according to the procedure described in Example 1, with the exception that tert-butyl 1,4-diazepane-1-carboxylate was replaced by tert-butyl 1-oxo-2,9-diazaspiro[5.5]undecan-9-carboxylate, which was first de-Boc under acidic condition before it was engaged in the reaction with A6. No de-Boc was needed after the coupling. Compound 21 was obtained as a yellow solid (160 mg, 35.5%).1 H NMR (400 MHz, DMSO-d6) d 8.66 (dd, J = 7.6, 4.8 Hz, 1H), 7.61 - 7.42 (m, 4H), 7.25 - 7.19 (m, 1H), 4.83 - 4.74 (m, 2H), 3.96 - 3.84 (m, 1H), 3.24 - 3.16 (m, 1H), 3.10 - 2.90 (m, 4H), 2.89 - 2.79 (m, 1H), 2.60 - 2.55 (m, 2H), 2.33 - 2.23 (m, 3H), 1.83 (t, J = 6.8 Hz, 1H), 1.70 - 1.61 (m, 1H), 1.53 - 1.42 (m, 1H), 1.33 - 1.24 (m, 1H), 1.16 (d, J = 2.7 Hz, 3H), 1.00 (d, J = 10.5 Hz, 3H), 0.62 (s, 1H). HRMS (ESI): m / z exact mass calculated for C 31 H 31 ClN4O4S [M+H] + 591.1833, found 591.1817.

[0105] Example 23

[0106] 3-((7-(5-chloro-3-methyl-2-(3-oxo-2,8-diazaspiro[4.5]decane-8-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 23) was prepared according to the procedure described in Reference Example 1, with the exception that tert-butyl 1,4-diazepane-1-carboxylate was replaced by 3-oxo-2,8-diazaspiro[4.5]decane-3-carboxylic acid (no need to remove Boc after coupling) to give Compound 23 as a grey solid (150 mg, 23.3% yield over two steps).

[0107] Reference Example 1, with the exception that tert-butyl 1,4-diazepane-1-carboxylate was replaced by 3-oxo-2,8-diazaspiro[4.5]decane-3-carboxylic acid (no need to remove Boc after coupling) to give Compound 23 as a grey solid (150 mg, 23.3% yield over two steps). 1H NMR (400 MHz, DMSO-d6) δ 8.68 (d, J = 4.8 Hz, 1H), 7.61 - 7.52 (m, 3H), 7.47 - 7.36 (m, 1H), 7.23 (t, 1H), 4.86 - 4.72 (m, 2H), 3.73 - 3.37 (m, 1H), 3.27 - 3.15 (m, 1H), 3.06 - 2.88 (m, 3H), 2.81 - 2.61 (m, 2H), 2.58 (s, 2H), 2.40 (d, J = 9.7 Hz, 1H), 2.27 (d, J = 4.1 Hz, 3H), 1.96 (d, J = 4.6 Hz, 1H), 1.79 - 1.44 (m, 1H), 1.42 - 1.34 (m, 1H), 1.16 (d, J = 3.3 Hz, 3H), 1.02 (d, J = 3.4 Hz, 3H), 0.91 - 0.64 (m, 1H). HRMS (ESI): m / z exact mass calculated for C 31 H 31 ClN4O4S[M+H] + 591.1833, found 591.1816.

[0108] Example 24

[0109] 1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2- b]pyridin-7-yl)-6-methylbenzoyl)azetidine-3-carboxamide (Compound 24).

[0110] Refer to Example 1, the only difference is that tert-butyl 1,4-diazepane-1-carboxylate is replaced by azetidine-3-carboxamide (no need to remove Boc after coupling), yellow solid of Compound 24 is prepared (10 mg, yield 1.2%). 1H NMR (400 MHz, DMSO-d6) δ 8.72 - 8.61 (m, 1H), 7.58 (d, J = 2.0 Hz, 1H), 7.56 - 7.50 (m, 2H), 7.35 (d, J = 15.8 Hz, 1H), 7.33 - 7.20 (m, 1H), 7.05 - 6.91 (m, 1H), 4.79 (d, J = 7.7 Hz, 2H), 3.88 - 3.79 (m, 1H), 3.70 - 3.62 (m, 1H), 3.61 - 3.52 (m, 1H), 3.49 - 3.35 (m, 1H), 3.27 - 2.88 (m, 1H), 2.59 (d, J = 4.8 Hz, 2H), 2.35 (d, J = 2.0 Hz, 3H), 1.16 (s, 3H), 0.99 (d, J = 3.4 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C 27 H 25 ClN4O4S[M+H] + 537.1363, found 537.1345.

[0111] Example 25

[0112] Preparation of 4-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3- yl)methyl)thieno[3,2-b]pyridin-7-yl)-6-methylbenzoyl)piperazine-1-carboxamide (Compound 25).

[0113] Compound 20 (0.311 mmol, 162 mg), trimethylsilyl isocyanate (0.373 mmol, 43 mg, 51 μL), triethylamine (0.404 mmol, 41 mg, 56 μL) were dissolved in tetrahydrofuran (2 mL) and stirred at room temperature for 3 hours. The reaction was monitored by TLC (dichloromethane:methanol = 15:1) and the solvent was removed by evaporation under reduced pressure. The residue was purified by preparative thin layer chromatography (dichloromethane:methanol = 20:1) to give Compound 25 as a white solid (67 mg, yield 38.1%). 1H NMR (400 MHz, DMSO-d6) δ 8.69 (d, J = 4.8 Hz, 1H), 7.60 (dd, J = 2.1, 0.8 Hz, 1H), 7.56 (d, J = 2.1 Hz, 1H), 7.54 (s, 1H), 7.21 (d, J = 4.8 Hz, 1H), 5.95 (s, 2H), 4.79 (s, 2H), 3.31 - 3.16 (m, 3H), 3.08 - 3.00 (m, 1H), 3.00 - 2.91 (m, 1H), 2.69 - 2.54 (m, 4H), 2.28 (s, 3H), 2.14 - 2.02 (m, 1H), 1.17 (s, 3H), 1.03 (s, 3H). HRMS (ESI): m / z exact mass calculated for C 28 H 28 ClN5O4S[M+H] + 566.1629, found 566.1608.

[0114] Example 26

[0115] Preparation of 4-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3- yl)methyl)thieno[3,2-b]pyridin-7-yl)-6-methylbenzoyl)-1,4-diazepane-1-carboxamide (Compound 26).

[0116] Following the procedure of Example 25, except substituting Compound 20 with Compound 1, white solid of Compound 26 was prepared (10 mg, 8.6% yield over two steps). 1 H NMR (400 MHz, DMSO-d6) δ 8.69 (d, J = 4.8 Hz, 1H), 7.60 (dd, J = 2.1, 0.8 Hz, 1H), 7.56 (d, J = 2.1 Hz, 1H), 7.54 (s, 1H), 7.21 (d, J = 4.8 Hz, 1H), 5.95 (s, 2H), 4.79 (s, 2H), 3.31 - 3.16 (m, 3H), 3.08 - 3.00 (m, 1H), 3.00 - 2.91 (m, 1H), 2.69 - 2.54 (m, 4H), 2.28 (s, 3H), 2.14 - 2.02 (m, 1H), 1.17 (s, 3H), 1.03 (s, 3H). HRMS (ESI): m / z exact mass calculated for C 29 H 30ClN5O4S[M+H] + 580.1785, found 580.1771.

[0117] Example 27

[0118] Preparation of 3-((7-(5-chloro-2-(4-(hydroxymethyl)piperidin-1-carbonyl)-3-methylphenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (compound 27).

[0119] Referring to Example 1, the only difference was that tert-butyl 1,4-diazacycloheptan-1-carboxylate was replaced with piperidine-4-methanol (no need for de-Boc after coupling), to obtain a brown solid of compound 27 (117 mg, yield 42.4%). 1 H NMR(400MHz, DMSO-d6)δ8.67(t,J=4.9Hz,1H),7.60–7.49(m,3H),7.21(dd,J=45.0,4.8H z,1H),4.79(t,J=3.6Hz,2H),4.26(s,1H),3.19–3.11(m,1H),3.05–2.92(m,1H),2.84–2 .73(m,1H),2.61–2.58(m,2H),2.46(dd,J=13.5,3.3Hz,1H),2.30(s,1H),2.23(s,2H),1 .53(d,J=12.9Hz,1H),1.43–1.21(m,3H),1.16(d,J=2.3Hz,3H),1.02(d,J=21.4Hz,3H).

[0120] Example 28

[0121] Preparation of 1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl)thieno[3,2-b]pyridin-7-yl)-6-methylbenzoyl)piperidine-4-carboxylic acid (compound 28).

[0122] Referring to Example 1, the only difference was that tert-butyl 1,4-diazacycloheptan-1-carboxylate was replaced with ethyl piperidine-4-carboxylate for coupling, followed by hydrolysis in concentrated hydrochloric acid / acetic acid (1 / 1) at 110°C for 3 hours. Finally, the mixture was purified by preparative TLC (dichloromethane:methanol = 15:1) to obtain a white solid of compound 28 (60 mg, two-step yield 7.6%). 1H NMR (400 MHz, DMSO-d6) δ 12.10 (s, 1H), 8.66 (d, J = 4.8 Hz, 1H), 7.60 - 7.50 (m, 3H), 7.26 - 7.15 (m, 1H), 4.83 - 4.72 (m, 2H), 4.05 (t, J = 15.1 Hz, 1H), 3.18 - 2.65 (m, 3H), 2.58 (d, J = 8.7 Hz, 2H), 2.46 - 2.13 (m, 5H), 1.68 - 1.57 (m, 1H), 1.42 - 1.24 (m, 2H), 1.16 (d, J = 1.8 Hz, 3H), 1.01 (s, 3H). HRMS (ESI): m / z exact mass calculated for C 29 H 28 ClN3O5S[M+H] + 566.1516, found 566.1500.

[0123] Example 29

[0124] Preparation of 3-((7-(5-chloro-3-methyl-2-(4-((methylamino)methyl)piperidine-1- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane- 2,4-dione (Compound 29).

[0125] Following the procedure of Example 1, but substituting tert-butyl methyl(piperidin-4- ylmethyl)carbamate for tert-butyl 1,4-diazepane-1-carboxylate, Compound 29 was prepared as a white solid (60 mg, 24.4% yield over two steps). 1 H NMR (400 MHz, DMSO-d6) δ 8.72 - 8.63 (m, 1H), 8.57 (s, 1H), 7.62 - 7.50 (m, 3H), 7.27 - 7.13 (m, 1H), 4.79 (d, J = 2.5 Hz, 2H), 4.25 - 4.17 (m, 1H), 3.20 - 2.97 (m, 1H), 2.92 - 2.81 (m, 0.5H), 2.76 - 2.66 (m, 1H), 2.64 - 2.60 (m, 2H), 2.58 - 2.53 (m, 0.5H), 2.46 - 2.37 (m, 3H), 2.34 - 2.12 (m, 5H), 1.76 - 1.56 (m, 2H), 1.52 - 1.27 (m, 1H), 1.25 - 1.10 (m, 4H), 1.10 - 0.93 (m, 4H). HRMS (ESI): m / z exact mass calculated for C 30 H33 CIN4O3S [M+H] + 565.2040, found 565.2023.

[0126] Example 30

[0127] Preparation of 3-((7-(5-chloro-2-(4-((dimethylamino)methyl)piperidine-l- carboxamido)-3-methylphenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 30).

[0128] Following Example 1, except that N,N-dimethyl-l-(piperidin-4-yl)methanamine hydrochloride was coupled instead of tert-butyl 1,4-diazepane-l-carboxylate without Boc removal after coupling, purple solid of Compound 30 was prepared (10 mg, 3.2% yield over two steps). 1 H NMR (400 MHz, DMSO) d 8.72 - 8.63 (m, 1H), 7.63 - 7.51 (m, 3H), 7.23 (d, J = 4.8 Hz, 1H), 4.80 (d, J = 2.6 Hz, 2H), 4.27 - 4.19 (m, 1H), 3.15 - 3.08 (m, 1H), 3.06 - 2.95 (m, 2H), 2.92 - 2.84 (m, 2H), 2.72 (s, 2H), 2.63 (d, J = 5.0 Hz, 6H), 2.43 (s, 1H), 2.34 - 2.22 (m, 3H), 2.11 - 1.87 (m, 1H), 1.84 - 1.68 (m, 2H), 1.68 - 1.49 (m, 1H), 1.18 (d, J = 3.0 Hz, 3H), 1.07 (s, 3H). HRMS (ESI): m / z exact mass calculated for C 31 H 35 CIN4O3S [M+H] + 579.2197, found 579.2180.

[0129] Example 31

[0130] Preparation of 3-((7-(5-chloro-2-(4-((dimethylamino)methyl)piperidine-l- carboxamido)-3-methylphenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 30).

[0131] Reference Example 1, except that tert-butyl 1,4-diazepane-1-carboxylate was replaced by N-methylhomopiperazine (no need to remove Boc after coupling) to give Compound 35 as a brown solid (12 mg, 3.6% yield over two steps). 1 H NMR (400 MHz, DMSO-d6) δ 8.72 - 8.63 (m, 1H), 7.63 - 7.51 (m, 3H), 7.23 (d, J = 4.8 Hz, 1H), 4.80 (d, J = 2.6 Hz, 2H), 4.27 - 4.19 (m, 1H), 3.15 - 3.08 (m, 1H), 3.06 - 2.95 (m, 2H), 2.92 - 2.84 (m, 2H), 2.72 (s, 2H), 2.63 (d, J = 5.0 Hz, 6H), 2.43 (s, 1H), 2.34 - 2.22 (m, 3H), 2.11 - 1.87 (m, 1H), 1.84 - 1.68 (m, 2H), 1.68 - 1.49 (m, 1H), 1.18 (d, J = 3.0 Hz, 3H), 1.07 (s, 3H).

[0132] Example 32

[0133] Preparation of 3-((7-(5-chloro-3-methyl-2-(morpholine-4-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 103).

[0134] Reference Example 1, except that tert-butyl 1,4-diazepane-1-carboxylate was replaced by morpholine (no need to remove Boc after coupling) to give Compound 103 as a brown solid (37 mg, 7% yield over two steps). 1 H NMR (400 MHz, DMSO-d6) δ 8.72 - 8.63 (m, 1H), 7.63 - 7.51 (m, 3H), 7.23 (d, J = 4.8 Hz, 1H), 4.80 (d, J = 2.6 Hz, 2H), 4.27 - 4.19 (m, 1H), 3.15 - 3.08 (m, 1H), 3.06 - 2.95 (m, 2H), 2.92 - 2.84 (m, 2H), 2.72 (s, 2H), 2.63 (d, J = 5.0 Hz, 6H), 2.43 (s, 1H), 2.34 - 2.22 (m, 3H), 2.11 - 1.87 (m, 1H), 1.84 - 1.68 (m, 2H), 1.68 - 1.49 (m, 1H), 1.18 (d, J = 3.0 Hz, 3H), 1.07 (s, 3H).

[0135] Example 33

[0136] 3-((7-(5-chloro-3-methyl-2-(4-methylpiperazine-l-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 104).

[0137] Following Example 1, except replacing tert-butyl 1,4-diazepane-l-carboxylate with N-methylpiperazine (no need to remove Boc after coupling), Compound 104 was prepared as a white solid (55 mg, 6% yield over two steps). 1 H NMR (300 MHz, DMSO-d6) δ 8.68 (d, J = 4.8 Hz, 1H), 7.63 - 7.54 (m, 2H), 7.56 (s, 1H), 7.56 (d, J = 4.8 Hz, 1H), 4.79 (d, J = 3.4 Hz, 2H), 3.41 - -3.09 (m, 1H), 3.03 - 2.88 (m, 1H), 2.72 - 2.63 (m, 1H), 2.65 - 2.55 (m, 2H), 2.24 (s, 3H), 2.24 - 2.17 (m, 1H), 1.96 - 1.87 (m, 1H), 1.85 (s, 3H), 1.46 - 1.31 (m, 1H), 1.17 (s, 3H), 1.03 (s, 3H), 0.81 (s, 1H).

[0138] Example 34

[0139] 3-((7-(5-chloro-3-methyl-2-(3-oxopiperazine-l-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 105).

[0140] Following Example 1, except replacing tert-butyl 1,4-diazepane-l-carboxylate with piperazin-2-one (no need to remove Boc after coupling), Compound 105 was prepared as a grey solid (45 mg, 0.4% yield over two steps). 1H NMR (300 MHz, DMSO-d6) δ 8.72 - 8.60 (m, 1H), 7.93 (d, J = 16.9 Hz, 2H), 7.66 - 7.49 (m, 3H), 4.88 - 4.74 (m, 2H), 3.69 - 3.55 (m, 1H), 3.51 (q, J = 6.7 Hz, 6.0 Hz, 1H), 3.38 - 3.30 (m, 3H), 3.25 - 2.95 (m, 1H), 2.93 - 2.80 (m, 1H), 2.63 - 2.57 (m, 2H), 2.27 (d, J = 7.2 Hz, 3H), 1.02 (d, J = 5.9 Hz, 3H).

[0141] Example 35

[0142] Preparation of 3-((7-(5-chloro-2-(2,8-diazaspiro[4.5]decane-8-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 106).

[0143] Referring to Example 8, except that 4-chloro-2-methylbenzoic acid was replaced by 4- chlorobenzoic acid, white solid of Compound 106 was prepared (260 mg, 36% yield over two steps). MS (ESI): m / z calculated for C 30 H 32 ClN4O3S[M+H] + 563.2, found 563.3.

[0144] Example 36

[0145] Preparation of 3-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2- b]pyridin-7-yl)-5-methyl-4-(2,6-diazaspiro[3.3]heptan-2-carbonyl)benzonitrile (Compound 107).

[0146] Referring to Example 12, except that 4-chloro-2-methylbenzoic acid was replaced by 4- cyano-2-methylbenzoic acid, white solid of Compound 107 was prepared (20 mg, 5% yield over two steps). 1H NMR (400 MHz, DMSO-d6) δ 8.71 (dd, J = 4.8, 0.7 Hz, 1H), 8.05 - 7.96 (m, 2H), 7.57 (d, J = 2.3 Hz, 1H), 7.32 - 7.22 (m, 1H), 4.82 - 4.77 (m, 2H), 3.76 (s, 1H), 3.73 - 3.58 (m, 1H), 3.50 - 3.42 (m, 1H), 3.23 - 3.14 (m, 1H), 3.11 - 2.93 (m, 2H), 2.58 (d, J = 3.1 Hz, 2H), 2.39 (d, J = 1.5 Hz, 3H), 2.18 - 2.00 (m, 1H), 1.95 - 1.64 (m, 2H), 1.16 (s, 3H), 1.03 (d, J = 1.1 Hz, 3H).

[0147] Example 37

[0148] Preparation of 4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3- yl)methyl)thieno[3,2-b]pyridin-7-yl)-6-methyl-N-(piperidin-4-yl)benzamide (Comparison Compound TSD-01-108).

[0149] Following Example 1, except replacing tert-butyl 1,4-diazepane-1-carboxylate with 1-(tert-butoxycarbonyl)-4-aminopiperidine, to afford light pink solid of compound TSD-01-108 (39 mg, 8.5% yield over three steps). 1 H NMR (300 MHz, DMSO-d6) δ 8.63 (d, J = 4.8 Hz, 1H), 8.37 (d, J = 7.9 Hz, 1H), 7.55 (d, J = 2.1 Hz, 1H), 7.50 (s, 1H), 7.45 (d, J = 2.1 Hz, 1H), 7.28 (d, J = 4.8 Hz, 1H), 4.78 (s, 2H), 3.57 (s, 3H), 2.88 (d, J = 12.6 Hz, 2H), 2.61 (s, 2H), 2.56 (s, 1H), 2.33 (s, 3H), 1.41 (d, J = 12.8 Hz, 2H), 1.18 (s, 3H), 1.15 - 1.08 (m, 2H), 1.04 (s, 3H).

[0150] Example 38

[0151] 4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2- b]pyridin-7-yl)-6-methyl-N-(piperidin-4-yl)benzamide (Comparative Compound WDL-04-123).

[0152] Referring to Example 1, except that tert-butyl 4-aminoazepane-1-carboxylate was used instead of tert-butyl 1,4-diazepane-1-carboxylate, white solid of Compound WDL-04-123 was prepared (22 mg, yield 4%). 1 H NMR (300 MHz, DMSO-d6) δ 8.64 (d, J = 4.8 Hz, 1H), 8.48 (d, J = 8.0 Hz, 2H), 7.56 (d, J = 2.1 Hz, 1H), 7.51 (s, 1H), 7.46 (d, J = 2.1 Hz, 1H), 7.27 (d, J = 4.7 Hz, 1H), 4.79 (s, 2H), 3.75 (s, 1H), 3.10 - 2.97 (m, 2H), 2.96 - 2.81 (m, 2H), 2.61 (s, 2H), 2.33 (s, 3H), 1.77 - 1.36 (m, 6H), 1.18 (s, 3H), 1.04 (s, 3H).

[0153] Example 39

[0154] 5-(1,4-diazepane-1-carbonyl)-4-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3- yl)methyl)thieno[3,2-b]pyridin-7-yl)-6-methylpyridinecarbonitrile (Comparative Compound WDL-06-173) was prepared.

[0155] (1) 4,6-dichloro-2-methylnicotinic acid (206 mg, 1.0 mmol), tert-butyl 1,4-diazepane-1- carboxylate (220 mg, 1.1 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (456 mg, 1.2 mmol), N,N-diisopropylethylamine (254 mg, 342 μL, 2.0 mmol) were dissolved in dry dichloromethane (5 mL) and stirred at room temperature overnight. The reaction was monitored to completion by TLC (petroleum ether: ethyl acetate = 1:1), the solvent was evaporated under reduced pressure and the residue was separated and purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to give intermediate A9 as a colorless transparent oil (508 mg, yield 71.0%).

[0156] (2) A9 (388 mg, 1 mmol), zinc cyanide (79 mg, 0.67 mmol), 1,1'- bis(diphenylphosphino)ferrocene (55 mg, 0.1 mmol), tris(dibenzylideneacetone)dipalladium (46 mg, 0.05 mmol) were dissolved in dry N,N-dimethylformamide (3 mL), replaced by argon for 3 times, heated at 100 °C for 12 hours. TLC (petroleum ether: ethyl acetate = 1:1) was used to monitor the reaction, the reaction solution was filtered by celite, extracted by ethyl acetate (40 mL x 3), the combined organic phase was washed by saturated sodium chloride aqueous solution (20 mL x 2), dried by anhydrous sodium sulfate, the solvent was removed by reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the yellow oil of intermediate A10 (205 mg, yield 54%).

[0157] (3) Refer to example 1, coupling of compound A10 with A5, and finally de-Boc to prepare the white solid of compound WDL-06-173 (8 mg, 3% yield for two steps). MS (ESI): m / z calculated for C 28 H 29 N6O3S[M+H] + 529.2, found 529.1.

[0158] Example 40

[0159] 4-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2- b]pyridin-7-yl)-6-methyl-5-(2,8-diazaspiro[4.5]decane-8-carbonyl)picolinonitrile (Comparison Compound WDL-06-183).

[0160] (1) 4,6-dichloro-2-methylnicotinic acid (588 mg, 2.85 mmol), 4-dimethylaminopyridine (70 mg, 0.57 mmol) were dissolved in tetrahydrofuran (10 mL), di-tert-butyl dicarbonate (1246 mg, 5.71 mmol) was added under stirring at room temperature. The reaction was carried out at room temperature overnight, TLC (petroleum ether: ethyl acetate = 1:1) was used to monitor the reaction, the solvent was removed by reduced pressure, the residue was purified by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the intermediate A12 (435 mg, yield 58%).

[0161] (2) A12 (435 mg, 1.66 mmol), zinc cyanide (130 mg, 1.11 mmol), 1,1'- bis(diphenylphosphino)ferrocene (92 mg, 0.17 mmol), tris(dibenzylideneacetone)dipalladium (76 mg, 0.083 mmol) were placed in dry N,N-dimethylformamide (5 mL), purged with argon for 3 times, heated at 100 °C for 12 h. TLC (petroleum ether: ethyl acetate = 10:1) was used to monitor the reaction. The reaction mixture was filtered through celite, and the filtrate was extracted with ethyl acetate (40 mL x 3), washed with saturated aqueous sodium chloride solution (20 mL x 2), dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 80:1) to give intermediate A13 as a yellow solid (188 mg, 45% yield).

[0162] (3) A13 (188 mg, 0.74 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (2 mL) was added. The mixture was stirred at room temperature overnight. TLC was used to monitor the reaction. The solvent was removed under reduced pressure to give A14 as a yellow solid (145 mg, 100% yield), which was used directly in the next step without further purification.

[0163] (4) A14 (145 mg, 0.74 mmol), 2,8-diazaspiro[4.5]decane-2-carboxylic acid tert-butyl ester (178 mg, 0.74 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (422 mg, 1.11 mmol), N,N-diisopropylethylamine (191 mg, 256 μL, 1.48 mmol) were dissolved in dry dichloromethane (3 mL), and the mixture was stirred at room temperature overnight. TLC (petroleum ether: ethyl acetate = 1:1) was used to monitor the reaction. The solvent was removed under reduced pressure. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to give intermediate A15 as a colorless transparent oil (300 mg, 96.8% yield).

[0164] (5) Referring to Example 1, compound A15 was coupled with A5, and finally deprotected to give compound WDL-06-183 as a white solid (30 mg, 7.5% yield for two steps). MS (ESI): m / z calculated for C 31 H 33 N6O3S[M+H] + 569.2, found 569.3.

[0165] Example 41

[0166] Preparation of ((1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3- azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2-b]pyridin-7-yl)-6- methylbenzoyl)piperidin-4-yl)methyl)isobutylcarbamate (Compound 108).

[0167] Referring to Example 1, except that tert-butyl N-(4-piperidinylmethyl)carbamate was used in place of tert-butyl 1,4-diazepane-1-carboxylate (no need to remove Boc after coupling), white solid of Compound 109 was obtained (4 mg, 2% yield over two steps). 1 H NMR (400 MHz, DMSO-d6) δ 8.68 (d, J = 4.8 Hz, 0.5H), 8.65 (d, J = 4.8 Hz, 0.5H), 7.58 - 7.52 (m, 3H), 7.23 (d, J = 4.8 Hz, 0.5H), 7.14 (d, J = 4.8 Hz, 0.5H), 6.83 - 6.73 (m, 0.5H), 6.66 - 6.61 (m, 0.5H), 4.79 (d, J = 2.6 Hz, 2H), 4.25 - 4.15 (m, 1H), 3.17 - 3.10 (m, 1H), 3.04 - 2.97 (m, 1H), 2.84 - 2.72 (m, 2H), 2.61 - 2.58 (m, 2H), 2.47 - 2.37 (m, 2H), 2.30 (s, 1H), 2.27 - 2.18 (m, 3H), 1.50 - 1.42 (m, 1H), 1.34 (d, J = 2.6 Hz, 9H), 1.31 - 1.22 (m, 2H), 1.17 (d, J = 2.6 Hz, 3H), 1.06 (d, J = 3.0 Hz, 2H), 0.98 (s, 1H). HRMS (ESI): m / z exact mass calculated for C 34 H 39 ClN4O5S[M+H] + 651.2408, found 651.2394.

[0168] Example 42

[0169] Preparation of ((1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3- azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2-b]pyridin-7-yl)-6- methylbenzoyl)piperidin-4-yl)methyl)isobutylcarbamate (Compound 108).

[0170] (1) 4-nitro-phenyl ester 1-chloro carboxylic acid ethyl ester (245 mg, 1.0 mmol), isobutyric acid (264 mg, 3.0 mmol) and silver oxide (232 mg, 1.0 mmol) were placed in a chicken heart bottle and heated with stirring at 90 °C overnight. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 5:1). The reaction was diluted with ethyl acetate (10 mL) and filtered through celite, and the filter cake was washed with ethyl acetate (5 mL x 2). The filtrate was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give intermediate D1 as a yellow oil (129 mg, yield 43%). The intermediate D1 was used directly in the next step without further purification. 1 H NMR (400 MHz, CDC13) δ 8.28 (d, J = 2.2 Hz, 1H), 8.26 (d, J = 2.1 Hz, 1H), 7.28 (d, J = 2.1 Hz, 1H), 7.26 (d, J = 1.4 Hz, 1H), 6.87 - 6.82 (m, 1H), 2.90 - 2.77 (m, 1H), 1.35 (s, 3H), 1.33 (s, 3H), 1.17 (d, J = 0.8 Hz, 1.5H), 1.15 (d, J = 0.8 Hz, 1.5H).

[0171] (2) Compound 17 (115 mg, 0.21 mmol), triethylamine (88 μL, 0.63 mmol) were dissolved in dichloromethane (2 mL), and D1 (93 mg, 0.31 mmol) was added, and stirred at room temperature overnight. The reaction was monitored by TLC (dichloromethane:methanol = 15:1). After the reaction was completed, the reaction was evaporated under reduced pressure, and the residue was purified by preparative thin layer chromatography (dichloromethane:methanol = 15:1), and the white solid of compound 108 was obtained by ether slurry, filtration and drying (70 mg, yield 47%). The compound 108 was used directly in the next step without further purification. 1H NMR (400 MHz, DMSO-d6) δ 8.68 (d, J = 4.8 Hz, 0.5H), 8.65 (d, J = 4.8 Hz, 0.5H), 7.71 - 7.64 (m, 0.5H), 7.60 - 7.58 (m, 0.5H), 7.57 (s, 1H), 7.55 - 7.50 (m, 2H), 7.25 (d, J = 4.8 Hz, 0.5H), 7.15 (d, J = 4.8 Hz, 0.5H), 4.79 (d, J = 2.6 Hz, 2H), 4.27 - 4.15 (m, 1H), 3.17 - 3.09 (m, 0.5H), 3.03 - 2.95 (m, 0.5H), 2.90 - 2.85 (m, 1H), 2.84 - 2.73 (m, 1H), 2.62 - 2.55 (m, 3H), 2.47 - 2.41 (m, 1H), 2.34 - 2.29 (m, 2H), 2.28 - 2.25 (m, 1H), 2.24 - 2.16 (m, 3H), 1.53 - 1.23 (m, 3H), 1.17 (d, J = 3.9 Hz, 3H), 1.06 (s, 1.5H), 0.99 (s, 1.5H), 0.95 - 0.94 (m, 3H), 0.94 - 0.92 (m, 3H). HRMS (ESI): m / z exact mass calculated for C 33 H 37 ClN4O4S[M+H] + 621.2302, found 621.2288.

[0172] Example 43

[0173] 3-((7-(5-chloro-3-methyl-2-(2-oxo-l-oxa-3,8-diazaspiro[4.5]decane-8- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 110)

[0174] Refer to Example 1, the only difference is that tert-butyl 2-oxo-l-oxa-3,8- diazaspiro[4.5]decane-8-carboxylate is used instead of tert-butyl 1,4- diazepane-l-carboxylate, and it is first de-Boc under acidic condition before it participates in the reaction with A6. No de-Boc is needed after coupling. Brown yellow solid of Compound 110 is obtained (4 mg, 2% yield for two steps). 1H NMR (400 MHz, DMSO-d6) δ 8.68 (d, J = 4.8 Hz, 0.5H), 8.65 (d, J = 4.8 Hz, 0.5H), 7.58 - 7.52 (m, 3H), 7.23 (d, J = 4.8 Hz, 0.5H), 7.14 (d, J = 4.8 Hz, 0.5H), 6.83 - 6.73 (m, 0.5H), 6.66 - 6.61 (m, 0.5H), 4.79 (d, J = 2.6 Hz, 2H), 4.25 - 4.15 (m, 1H), 3.17 - 3.10 (m, 1H), 3.04 - 2.97 (m, 1H), 2.84 - 2.72 (m, 2H), 2.61 - 2.58 (m, 2H), 2.47 - 2.37 (m, 2H), 2.30 (s, 1H), 2.27 - 2.18 (m, 3H), 1.50 - 1.42 (m, 1H), 1.34 (d, J = 2.6 Hz, 9H), 1.31 - 1.22 (m, 2H), 1.17 (d, J = 2.6 Hz, 3H), 1.06 (d, J = 3.0 Hz, 2H), 0.98 (s, 1H). HRMS (ESI): m / z exact mass calculated for C 34 H 39 ClN4O5S[M+H] + 651.2408, found 651.2394.

[0175] Example 44

[0176] Preparation of 3-((7-(2-(4-(1-aminoethyl)piperidine-1-carbonyl)-5-chloro-3- methylphenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane- 2,4-dione (Compound 111).

[0177] (1) 4-(1-aminoethyl)-1-piperidinecarboxylic acid-1,1-dimethylethyl ester (200 mg, 0.88 mmol), potassium carbonate (363 mg, 2.63 mmol) were dissolved in a mixture of ether / water (3 / 1) (4 mL), and benzyl chloroformate (370 μL, 2.63 mmol) was added with stirring in an ice bath. The reaction was allowed to proceed at room temperature overnight, and TLC (dichloromethane:methanol = 30:1) was used to monitor the completion of the reaction. The reaction was quenched with saturated sodium bicarbonate solution, and extracted with dichloromethane (20 mL x 2). The organic phase was combined and washed with saturated aqueous sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane:methanol = 90:1) to give intermediate El as colorless oil (317 mg, yield 100%). The structure of intermediate El was confirmed by1H NMR. 1H NMR (400 MHz, CDC13) δ 7.36 (s, 5H), 5.18 - 4.98 (m, 2H), 4.27 - 3.97 (m, 2H), 3.77 - 3.53 (m, 1H), 2.70 - 2.54 (m, 2H), 1.60 (d, J = 7.3 Hz, 1H), 1.45 (s, 9H), 1.34 - 1.14 (m, 4H), 1.11 (d, J = 6.8 Hz, 3H).

[0178] (2) Dissolve E1 (317 mg, 0.88 mmol) in dichloromethane (3 mL), add hydrogen chloride in ethyl acetate (4 N, 2.2 mL) at 0 °C, stir at room temperature for 3 h. TLC (dichloromethane:methanol = 30:1) monitor the reaction is completed, directly remove the solvent under reduced pressure to obtain E2 white solid directly into the next step.

[0179] (3) Refer to Example 1, the difference is only 1,4-diazepane-1-carboxylic acid tert-butyl ester is replaced by intermediate E2, after the amide condensation, coupling to prepare E4 brown yellow foam (230 mg, 38%) solid.

[0180] (4) Dissolve E4 (230 mg, 0.33 mmol) in acetic acid (0.6 mL), replace argon 3 times, inject 33% aqueous hydrogen bromide solution (1 mL), stir at room temperature for 3 h. TLC (dichloromethane:methanol = 15:1) monitor the reaction is completed, add saturated sodium bicarbonate solution to quench the reaction. Extracted with dichloromethane (20 mL x 2), combined organic phase, washed with saturated aqueous sodium chloride (10 mL), dried over anhydrous sodium sulfate, remove the solvent under reduced pressure, the residue was purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to give the crude product. The crude product was further purified by preparative thin layer chromatography (dichloromethane:methanol = 15:1), ether, suction filtration and drying to give compound 111 white solid (50 mg, yield 27%). Compound 111 1H NMR (400 MHz, DMSO-d6) δ 8.67 (d, J = 4.8 Hz, 0.5H), 8.64 (d, J = 4.8 Hz, 0.5H), 7.59 - 7.35 (m, 4H), 7.25 (dd, J = 4.8, 2.4 Hz, 0.5H), 7.13 (d, J = 4.8 Hz, 0.5H), 4.85 - 4.71 (m, 2H), 4.32 (m, 1H), 3.21 - 3.13 (m, 0.5H), 3.04 - 2.93 (m, 1H), 2.88 - 2.76 (m, 1H), 2.73 - 2.66 (m, 0.5H), 2.59 (d, J = 1.6 Hz, 1H), 2.57 (d, J = 1.6 Hz, 1H), 2.54 (s, 1H), 2.47 - 2.37 (m, 1H), 2.34 - 2.06 (m, 4H), 1.64 - 1.38 (m, 2.5H), 1.18 - 1.14 (m, 3H), 1.10 - 1.03 (m, 3.5H), 0.97 (d, J = 3.5 Hz, 1H), 0.69 (d, J = 6.7 Hz, 1H), 0.51 (d, J = 6.7 Hz, 1H). HRMS (ESI): m / z exact mass calculated for C 30 H 33 ClN4O3S[M+H] + 565.2040, found 565.2029.

[0181] Example 45

[0182] Preparation of N-((1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3- azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2-b]pyridin-7-yl)-6- methylbenzoyl)piperidin-4-yl)methyl)isobutyramide (Compound 112)

[0183] Compound 17 (253 mg, 0.46 mmol), triethylamine (128 μL, 0.92 mmol) were dissolved in dichloromethane (1 mL), diethylpyrocarbonate (80 μL, 0.55 mmol) was added under ice-bath stirring. After 4 h stirring at room temperature, the reaction was monitored by TLC (dichloromethane:methanol = 30:1) and the solvent was removed by evaporation under reduced pressure. The residue was purified by preparative thin layer chromatography (dichloromethane:methanol = 15:1) and the product was washed with n-hexane, filtered and dried to give Compound 112 as a light yellow solid (141 mg, yield 49%). Compound 112 1H NMR (400 MHz, DMSO-d6) δ 8.67 (d, J = 4.8 Hz, 0.5H), 8.65 (d, J = 4.8 Hz, 0.5H), 7.60 - 7.51 (m, 3H), 7.24 (d, J = 4.8 Hz, 0.5H), 7.14 (d, J = 4.8 Hz, 0.5H), 7.07 (t, J = 5.8 Hz, 0.5H), 6.91 (t, J = 5.8 Hz, 0.5H), 4.84 - 4.74 (m, 2H), 4.28 - 4.14 (m, 1H), 3.92 (q, J = 7.1 Hz, 2H), 3.17 - 3.09 (m, 0.5H), 3.02 - 2.94 (m, 1.5H), 2.83 - 2.76 (m, 1H), 2.63 - 2.55 (m, 2H), 2.47 - 2.41 (m, 1H), 2.30 (s, 1H), 2.26 - 2.17 (m, 3H), 1.52 - 1.43 (m, 1H), 1.35 - 1.21 (m, 3H), 1.18 - 1.16 (m, 4H), 1.13 - 1.11 (m, 3H), 1.06 (s, 1.5H), 0.98 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 32 H 35 ClN4O5S[M + H] + 623.2095, found 623.2080.

[0184] Example 46

[0185] Preparation of N-((1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3- azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2-b]pyridin-7-yl)-6- methylbenzoyl)piperidin-4-yl)methyl)isobutyramide (Compound 113).

[0186] Refer to Example 45, the only difference is to replace diethylpyrocarbonate with dimethyl dicarbonate to produce Compound 113 as a white solid (40 mg, yield 45%). The data of Compound 113 1H NMR (400 MHz, DMSO-d6) δ 8.67 (d, J = 4.8 Hz, 0.5H), 8.65 (d, J = 4.8 Hz, 0.5H), 7.61 - 7.50 (m, 3H), 7.24 (d, J = 4.8 Hz, 0.5H), 7.17 - 7.08 (m, 1H), 6.96 (t, J = 5.9 Hz, 0.5H), 4.79 (d, J = 2.7 Hz, 2H), 4.28 - 4.14 (m, 1H), 3.47 (d, J = 2.1 Hz, 3H), 3.21 - 3.15 (m, 1H), 3.05 - 2.89 (m, 1H), 2.84 - 2.73 (m, 1H), 2.64 - 2.55 (m, 2H), 2.48 - 2.40 (m, 1H), 2.30 (s, 1H), 2.22 (s, 3H), 1.54 - 1.42 (m, 1H), 1.30 - 1.25 (m, 2H), 1.23 (s, 2H), 1.17 (d, J = 4.6 Hz, 3H), 1.06 (s, 1.5H), 0.98 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 31 H 33 ClN4O5S[M+H] + 609.1938, found 609.1925.

[0187] Example 47

[0188] Preparation of N-((1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3- azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2-b]pyridin-7-yl)-6- methylbenzoyl)piperidin-4-yl)methyl)isobutyramide (Compound 114).

[0189] Referring to Example 45, except that diethylpyrocarbonate was replaced by isopropyl chloroformate, Compound 114 was prepared as a white solid (65 mg, yield 55%). The data of Compound 114: 1H NMR (400 MHz, DMSO-d6) 8.67 (d, J = 4.8 Hz, 0.5H), 8.65 (d, J = 4.8 Hz, 0.5H),, 7.61 - 7.49 (m, 3H), 7.24 (d, J = 4.8 Hz, 0.5H), 7.15 (d, J = 4.8 Hz, 0.5H), 7.00 (t, J = 5.9 Hz, 0.5H), 6.85 (t, J = 5.9 Hz, 0.5H), 4.79 (d, J = 2.4 Hz, 2H), 4.74 - 4.63 (m, 1H), 4.28 - 4.13 (m, 1H), 3.13 (d, J = 13.1 Hz, 0.5H), 2.99 (d, J = 13.2 Hz, 0.5H), 2.83 - 2.76 (m, 1H), 2.65 - 2.55 (m, 2H), 2.48 - 2.40 (m, 1H), 2.30 (s, 1H), 2.22 (s, 3H), 1.47 (s, 1H), 1.34 (d, J = 6.1 Hz, 1H), 1.31 - 1.28 (m, 1H), 1.25 (d, J = 3.8 Hz, 1H), 1.23 (s, 2H), 1.17 (d, J = 3.3 Hz, 3H), 1.12 (dd, J = 6.3, 2.8 Hz, 6H), 1.06 (s, 1.5H), 0.98 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 33 H 37 ClN4O5S[M+H] + 637.2251, found 637.2239.

[0190] Example 48

[0191] Preparation of N-((1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3- azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2-b]pyridin-7-yl)-6- methylbenzoyl)piperidin-4-yl)methyl)acetamide (Compound 115).

[0192] Following the procedure of Example 45, except substituting acetyl chloride for diethylpyrocarbonate, Compound 115 was prepared as a white foamy solid (70 mg, 65% yield). Compound 115 1H NMR (400 MHz, DMSO-d6) δ 8.68 (d, J = 4.8 Hz, 0.5H), 8.65 (d, J = 4.8 Hz, 0.5H), 7.78 (d, J = 5.8 Hz, 0.5H), 7.61 - 7.57 (m, 2H), 7.55 - 7.51 (m, 1.5H), 7.25 (d, J = 4.8 Hz, 0.5H), 7.15 (d, J = 4.8 Hz, 0.5H), 4.79 (s, 2H), 4.28 - 4.22 (m, 0.5H), 4.21 - 4.15 (m, 0.5H), 3.17 - 3.09 (m, 0.5H), 3.02 - 2.94 (m, 0.5H), 2.89 - 2.83 (m, 1H), 2.83 - 2.73 (m, 1H), 2.61 - 2.59 (m, 2H), 2.57 - 2.53 (m, 0.5H), 2.47 - 2.41 (m, 0.5H), 2.33 - 2.29 (m, 1.5H), 2.27 - 2.17 (m, 3.5H), 1.74 (d, J = 3.4 Hz, 3H), 1.53 - 1.45 (m, 1H), 1.41 - 1.21 (m, 3H), 1.17 (d, J = 3.1 Hz, 3H), 1.06 (s, 1.5H), 0.99 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 31 H 33 CIN4O4S [M+H] + 593.1989, found 593.1973.

[0193] Example 49

[0194] 3-((7-(5-chloro-3-methyl-2-(4-((pyrimidin-2-ylamino)methyl)piperidine-l- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 116)

[0195] Compound 17 (50 mg, 0.091 mmol) was dissolved in ethanol (1 mL) in a sealed tube, diisopropylethylamine (24 μL, 0.14 mmol) and 2-chloropyrimidine (58 mg, 0.11 mmol) were added, and heating stirring was performed at 90 °C. After cooling to room temperature, the reaction was monitored by TLC (dichloromethane:methanol = 15:1). After the reaction was completed, ethanol was evaporated under reduced pressure, and the residue was purified by preparative thin layer chromatography (dichloromethane:methanol = 15:1), and Compound 116 was obtained as a white solid (19 mg, yield 33%) by slurry in ethyl ether, suction filtration and drying. Compound 116 1H NMR(400MHz, DMSO-d6)δ8.69(d,J=4.8Hz,0.5H),8.65(d,J=4.8Hz,0.5H)),8.23–8.16(m,2H),7.60–7.49(m,3H),7.25(d, J=4.8Hz,0.5H),7.15(t,J=5.1Hz,1H),6.98(t,J=5.9Hz,0.5H),6.50(q,J=5.0Hz,1H),4.78(d,J=2.3Hz,2H),4.28–4.13( m,1H),3.18–2.95(m,2H),2.84–2.70(m,1H),2.63–2.55(m,2H),2.48–2.41(m,1H),2.36–2.17(m,4H),1.59–1.44(m,2H), 1.40–1.27(m,1H),1.26–1.19(m,1H),1.14(d,J=10.5Hz,3H),1.04(s,2H),0.95(s,1H),0.93–0.79(m,1H).HRMS(ESI):m / z exact mass calculated for C 33 H 33 ClN6O3S[M+H] + 629.2102, found 629.2085.

[0196] Example 50

[0197] Preparation of 1-((1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl)thiophene[3,2-b]pyridin-7-yl)-6-methylbenzoyl)piperidin-4-yl)methyl)urea (compound 117).

[0198] Referring to Example 25, the only difference being that compound 20 was replaced with compound 17, yielding compound 117 as a white solid (183 mg, yield 57%). Compound 117... 1H NMR (400 MHz, DMSO-d6) δ 8.68 (d, J = 4.8 Hz, 0.5H), 8.65 (d, J = 4.8 Hz, 0.5H), 7.59 - 7.51 (m, 3H), 7.25 (d, J = 4.8 Hz, 0.5H), 7.15 (d, J = 4.8 Hz, 0.5H), 5.93 (t, J = 5.9 Hz, 0.5H), 5.80 (t, J = 5.8 Hz, 0.5H), 5.33 (s, 1H), 5.31 (s, 1H), 4.84 - 4.74 (m, 2H), 4.28 - 4.13 (m, 1H), 3.19 - 3.09 (m, 0.5H), 3.04 - 2.97 (m, 0.5H), 2.88 - 2.71 (m, 2H), 2.60 (s, 1H), 2.59 (s, 1H), 2.48 - 2.42 (m, 1H), 2.33 - 2.13 (m, 5H), 1.51 - 1.43 (m, 1H), 1.36 - 1.22 (m, 2H), 1.17 (d, J = 2.8 Hz, 3H), 1.06 (s, 1.5H), 0.99 (s, 1.5H), 0.95 - 0.81 (m, 1H). HRMS (ESI): m / z exact mass calculated for C 30 H 32 ClN5O4S[M+H] + 594.1942, found 594.1926.

[0199] Example 51

[0200] 1-((1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2- b]pyridin-7-yl)-6-methylbenzoyl)piperidin-4-yl)-3-ethyl)urea (Compound 118).

[0201] Following the procedure of Example 50, except substituting ethyl isocyanate for trimethylsilyl isocyanate, Compound 118 was prepared as a white solid (50 mg, 44% yield). Compound 118 1H NMR (400MHz, DMSO-d6).δ8.68(d,J=4.8Hz,0.5H),8.65(d,J=4.8Hz,0.5H),7.60–7.50(m,3H),7.25(d,J=4.8Hz,0.5H),7.15(d,J =4.8Hz,0.5H),5.85(t,J=5.9Hz,0.5H),5.77–5.66(m,1.5H),4.79(d,J=1.8Hz,2H),4.28–4.14(m,1H),3.19–3.10(m,0.5H),3.0 3–2.89(m,2.5H),2.84–2.75(m,1H),2.64–2.56(m,2H),2.55–2.51(m,1H),2.48–2.42(m,1H),2.40–2.10(m,5H),1.51–1.43(m,1 H),1.40–1.23(m,2H),1.17(d,J=3.0Hz,3H),1.05(s,1.5H),0.99(s,1.5H),0.96–0.91(m,3H),0.90–0.76(m,1H).HRMS(ESI):m / z exact mass calculated for C 32 H 36 ClN5O4S[M+H] + 622.2255, found 622.2238.

[0202] Example 52

[0203] Preparation of 3-((7-(2-(4-(aminomethyl)-4-fluoropiperidine-1-carbonyl)-5-chloro-3-methylphenyl)thiophene[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (compound 121).

[0204] Referring to Example 1, the only difference was that tert-butyl 1,4-diazacycloheptan-1-carboxylate was replaced with N-((4-fluoropiperidin-4-yl)methyl)carbamate tert-butyl ester to prepare compound 121 as a white solid (26 mg, 9% yield in three steps). Compound 121... 1H NMR (400 MHz, DMSO-d6) δ 8.72 - 8.64 (m, 1H), 7.63 - 7.51 (m, 3H), 7.23 (d, J = 4.8 Hz, 0.5H), 7.17 (d, J = 4.8 Hz, 0.5H), 6.24 (s, 2H), 4.79 (d, J = 2.8 Hz, 2H), 4.13 - 3.98 (m, 1H), 3.13 - 3.05 (m, 1H), 3.00 - 2.89 (m, 1H), 2.88 - 2.80 (m, 1H), 2.61 (s, 1H), 2.59 (s, 1H), 2.48 - 2.37 (m, 1H), 2.34 - 2.15 (m, 4H), 1.77 - 1.51 (m, 2H), 1.50 - 1.22 (m, 2H), 1.17 (d, J = 5.0 Hz, 3H), 1.06 (s, 1.5H), 0.98 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 29 H 30 ClFN4O3S [M+H] + 569.1789, found 569.1773.

[0205] Example 53

[0206] Preparation of 3-((7-(5-chloro-2-(6,6-difluoro-l,4- diazepane-l-carbonyl)-3-methylphenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6- dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 92).

[0207] Following the procedure of Example 1, except substituting 6,6-difluoro-l,4-diaza- 1-carboxylic acid tert-butyl ester for 1,4-diazepane-l-carboxylic acid tert-butyl ester, Compound 92 was prepared as a white solid (10 mg, 4% yield over three steps). Compound 121 1H NMR (400 MHz, DMSO-d6) δ 8.65 - 8.57 (m, 1H), 7.60 - 7.53 (m, 2H), 7.51 (s, 1H), 7.29 - 7.20 (m, 1H), 4.77 (s, 2H), 4.14 - 3.99 (m, 1H), 3.68 - 3.55 (m, 1H), 3.41 - 3.31 (m, 1H), 3.03 (s, 1H), 2.91 - 2.74 (m, 2H), 2.54 (d, J = 2.0 Hz, 2H), 2.25 (d, J = 8.2 Hz, 3H), 1.93 - 1.76 (m, 1H), 1.74 - 1.59 (m, 1H), 1.12 (d, J = 6.0 Hz, 3H), 1.00 - 0.90 (m, 3H). HRMS (ESI): m / z exact mass calculated for C 28 H 27 ClF2N4O3S[M+H] + 573.1539, found 573.1527.

[0208] Example 54

[0209] 3-((7-(2-(4-(Aminomethyl)piperidine-l-carbonyl)-5-chloro-3-(methoxy-d3)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 119).

[0210] (1) 4-Chloro-2-hydroxybenzoic acid methyl ester (200 mg, 1.07 mmol), potassium carbonate (222 mg, 1.61 mmol) were dissolved in acetone (3 mL), and deuterated methyl iodide (80 μL, 1.3 mmol) was added slowly. The reaction was stirred at room temperature for 6 h, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 10: 1). The reaction was quenched by adding 10 mL of water, and the aqueous phase was extracted with ethyl acetate (20 mL x 2). The combined organic phase was washed with saturated sodium chloride solution (10 mL x 2) and dried over anhydrous sodium sulfate. The organic solvent was removed by evaporation under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 20: 1) to obtain intermediate Fl as a yellow oil (207 mg, yield 95%). The intermediate Fl was used directly in the next step without further purification. 1 H NMR (400 MHz, CDCl3) δ 7.79 - 7.72 (m, 1H), 7.00 - 6.93 (m, 2H), 3.88 (s, 3H).

[0211] (2) F1 (207 mg, 1.02 mmol) was dissolved in a mixture of methanol / water (1 / 1) (3 mL), and potassium hydroxide (286 mg, 5.10 mmol) was added under ice-bath stirring. The reaction was stirred at room temperature for 6 h, and TLC (petroleum ether: ethyl acetate = 10:1) was used to monitor the completion of the reaction. After the reaction was completed, 15 mL of water was added, and the aqueous phase was washed with dichloromethane (10 mL). The aqueous phase was retained, and the organic phase was discarded. Hydrochloric acid (1 N, 10 mL) was added to the aqueous phase to adjust the pH to 1-2, and the aqueous phase was extracted with dichloromethane (20 mL x 2). The combined organic phase was washed with saturated sodium chloride solution (10 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude intermediate F2 as a yellow oil, which was directly used in the next step.

[0212] (3) Intermediate F2, diacetoxyiodobenzene (656 mg, 2.04 mmol), iodine (517 mg, 2.04 mmol), and palladium acetate (12 mg, 0.051 mmol) were added to anhydrous N,N-dimethylformamide (3 mL), and the mixture was replaced with argon three times. The mixture was heated to 80°C and stirred for 8 h. TLC (petroleum ether: ethyl acetate = 1:1, with 1 drop of acetic acid) was used to monitor the completion of the reaction. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with methyl tert-butyl ether (5 mL), and then quenched with sodium metabisulfite. Concentrated hydrochloric acid was added to adjust the pH to 1, and the mixture was extracted with methyl tert-butyl ether (20 mL x 3). The combined organic phase was adjusted to pH 14 with saturated aqueous sodium hydroxide at 0°C, and the organic phase was discarded. The aqueous phase was adjusted to pH 1 with concentrated hydrochloric acid at 0°C, and then extracted with methyl tert-butyl ether (20 mL x 3). The solvent was removed under reduced pressure to obtain intermediate F3 as a white solid (254 mg, 79% yield over two steps). 1 H NMR (400 MHz, CDCl3) δ 10.75-9.55 (s, 1H), δ 8.58 (s, 1H), 7.17 (s, 1H).

[0213] (4) Refer to Example 17, except that intermediate A6 was replaced with intermediate F3, to obtain the target compound 119 as a light yellow solid (70 mg, 42% yield over two steps). 1H NMR (400 MHz, DMSO-d6, δ 8.75 - 8.68 (m, 1H), 7.81 (s, 2H), 7.56 - 7.51 (m, 1H), 7.42 - 7.39 (m, 1H), 7.35 - 7.28 (m, 2H), 4.78 (s, 2H), 4.50 - 4.43 (m, 1H), 3.09 - 2.91 (m, 2H), 2.79 - 2.65 (m, 3H), 2.62 - 2.58 (m, 1H), 1.88 - 1.61 (m, 3H), 1.26 - 1.21 (m, 1H), 1.17 (s, 3H), 1.15 - 1.06 (m, 2H), 1.01 (s, 3H). HRMS (ESI): m / z exact mass calculated for C 28 H 28 D3ClN4O4S[M+H] + 570.2021, found 570.2007.

[0214] Example 55

[0215] 3-((7-(2-(4-(Aminomethyl)piperidine-l-carbonyl)-3,5-dichlorophenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 120).

[0216] Following the procedure of Example 17, except substituting 2,4-dichlorobenzoic acid for 4-chloro-2-methylbenzoic acid, Compound 120 was prepared as a white solid (56 mg, 21% yield over four steps). Compound 120 was characterized by the following data: 1H NMR (400 MHz, DMSO-d6) δ 8.73 (d, J = 4.8 Hz, 0.5H), 8.68 (d, J = 4.8 Hz, 0.5H), 7.97 - 7.65 (m, 4H), 7.55 (d, J = 5.5 Hz, 1H), 7.25 (d, J = 4.8 Hz, 0.5H), 7.17 (d, J = 4.8 Hz, 0.5H), 4.80 (d, J = 2.7 Hz, 2H), 4.22 - 4.14 (m, 1H), 3.23 - 3.15 (m, 0.5H), 2.99 - 2.89 (m, 0.5H), 2.68 - 2.54 (m, 4H), 2.47 - 2.29 (m, 1.5H), 2.20 - 2.08 (m, 0.5H), 1.72 - 1.43 (m, 3H), 1.34 - 1.12 (m, 5H), 1.07 (s, 1.5H), 1.03 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 28 H 28 Cl2N4O3S [M+H] + 571.1337, found 571.1323.

[0217] Example 56

[0218] 3-((7-(2-(3-(Aminomethyl)azetidine-l-carbonyl)-3,5-dichlorophenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 123).

[0219] Following the procedure of Example 18, except substituting 2,4-dichlorobenzoic acid for 4-chloro-2-methylbenzoic acid, Compound 123 was prepared as a white solid (27 mg, 11% yield over four steps). Compound 123 1H NMR (400 MHz, DMSO-d6) δ 8.76 - 8.67 (m, 1H), 7.98 - 7.90 (m, 1H), 7.76 - 7.70 (m, 1H), 7.55 (s, 1H), 7.28 (d, J = 4.8 Hz, 0.5H), 7.25 (d, J = 4.8 Hz, 0.5H), 4.79 (s, 2H), 3.99 - 3.90 (m, 1H), 3.70 - 3.66 (m, 1H), 3.66 - 3.61 (m, 2H), 3.40 - 3.32 (m, 1H), 2.93 - 2.86 (m, 1H), 2.61 - 2.55 (m, 3H), 1.16 (s, 3H), 1.01 (d, J = 6.3 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C 26 H 24 Cl2N4O3S[M+H] + 543.1024, found 543.1010.

[0220] Example 57

[0221] 3-((7-(2-(4-(Aminomethyl)-4-fluoropiperidine-1-carbonyl)-3,5-dichlorophenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 125)

[0222] Refer to Example 52, except replace 4-chloro-2-methylbenzoic acid with 2,4- dichlorobenzoic acid reaction to produce Compound 125 as a light yellow solid (19 mg, 8% yield over four steps). Compound 125 1 H NMR (400 MHz, DMSO-d6) δ 8.70 (m, 1H), 7.96 (s, 1H), 7.81 (s, 0.5H), 7.76 (s, 0.5H), 7.57 - 7.52 (m, 1H), 7.24 (d, J = 4.8 Hz, 0.5H), 7.18 (d, J = 4.8 Hz, 0.5H), 4.80 (d, J = 2.9 Hz, 4H), 4.05 - 3.98 (m, 1H), 3.28 - 3.18 (m, 1H), 3.16 - 3.09 (m, 1H), 2.95 - 2.72 (m, 2H), 2.68 - 2.57 (m, 3H), 1.77 - 1.34 (m, 4H), 1.17 (d, J = 4.6 Hz, 3H), 1.07 (s, 1.5H), 1.01 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C28 H 27 Cl2FN4O3S[M+H] + 589.1243, found 589.1228.

[0223] Example 58

[0224] Preparation of 3-((7-(3,5-dichloro-2-(3-(((2,2,2-trifluoroethylamino)methyl)azetidine-1- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4- dione (Compound 134).

[0225] Compound 123 (60 mg, 0.11 mmol), triethylamine (46 μL, 0.33 mmol) were dissolved in tetrahydrofuran (1 mL), 2,2,2-trifluoroethyl trifluoromethanesulfonate (32 μL, 0.22 mmol) was added under ice-bath stirring, and the reaction was allowed to proceed at room temperature overnight. After the reaction was completed as monitored by TLC (dichloromethane:methanol = 20:1), the reaction was quenched by adding water (10 mL), extracted with dichloromethane (10 mL x 2), washed with saturated sodium chloride solution (10 mL), and dried over anhydrous sodium sulfate. The organic solvent was evaporated under reduced pressure, and the residue was purified by preparative thin layer chromatography, washed with n-hexane, and dried by suction filtration to obtain Compound 134 as a white solid (45 mg, yield 65%). Compound 134 1 H NMR (400 MHz, DMSO-d6) δ 8.72 (d, J = 2.1 Hz, 0.5H), 8.71 (d, J = 2.1 Hz, 0.5H), 7.96 (d, J = 1.9 Hz, 0.5H), 7.95 (d, J = 1.9 Hz, 0.5H), 7.78 (d, J = 1.9 Hz, 0.5H), 7.75 (d, J = 1.9 Hz, 0.5H), 7.57 (d, J = 1.0 Hz, 1H), 7.26 (t, J = 4.8 Hz, 1H), 4.83 - 4.78 (m, 2H), 3.93 - 3.80 (m, 1H), 3.66 - 3.50 (m, 2H), 3.24 - 3.15 (m, 2H), 3.14 - 2.93 (m, 2H), 2.73 - 2.69 (m, 1H), 2.59 (s, 2H), 2.34 - 2.05 (m, 2H), 1.16 (d, J = 5.0 Hz, 3H), 1.02 (s, 1.5H), 0.99 (s, 1.5H).. HRMS (ESI): m / z exact mass calculated for C 28 H 25 Cl2F3N4O3S[M+H] +625.1055, found 625.1042.

[0226] Example 59

[0227] N-((1-(2,4-dichloro-6-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3- yl)methyl)thieno[3,2-b]pyridin-7-yl)benzoyl)azetidin-3-yl)methyl)-2,2,2- trifluoroacetamide (Compound 135).

[0228] Referring to Example 58, the only difference is that 2,2,2-trifluoroethyl trifluoromethanesulfonate is replaced by trifluoroacetyl trifluoromethanesulfonate reaction to prepare Compound 135 as a white solid (44 mg, yield 62%). Compound 135 1 H NMR (400 MHz, DMSO-d6) δ 9.57 (t, J = 5.8 Hz, 0.5H), 9.47 (t, J = 5.8 Hz, 0.5H), 8.73 (d, J = 4.8 Hz, 0.5H), 8.71 (d, J = 4.8 Hz, 0.5H), 7.98 (d, J = 2.0 Hz, 0.5H), 7.96 (d, J = 2.0 Hz, 0.5H), 7.76 (t, J = 1.9 Hz, 1H), 7.56 (s, 1H), 7.27 (d, J = 4.8 Hz, 0.5H), 7.25 (d, J = 4.8 Hz, 0.5H), 4.80 (d, J = 3.4 Hz, 2H), 3.96 (q, J = 8.9 Hz, 1H), 3.66 (d, J = 7.0 Hz, 2H), 3.61 - 3.53 (m, 0.5H), 3.41 - 3.36 (m, 1H), 3.17 - 3.06 (m, 0.5H), 2.92 - 2.80 (m, 0.5H), 2.59 (d, J = 1.8 Hz, 3.5H), 1.16 (d, J = 3.7 Hz, 3H), 1.03 (s, 1.5H), 1.00 (s, 1.5H).. HRMS (ESI): m / z exact mass calculated for C 28 H 23 Cl2F3N4O4S [M+H] + 625.1055, found 639.0839.

[0229] Example 60

[0230] Preparation of N-((1-(2,4-dichloro-6-(2-((6,6-dimethyl-2,4-dioxo-3- azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2-b]pyridin-7-yl)benzoyl)azetidin-3- yl)methyl)-2,2-difluoroacetamide (Compound 140).

[0231] Following the procedure of Example 58, except replacing 2,2,2-trifluoroethyl triflate with difluoroacetyl triflate, Compound 140 was prepared as a white solid (45 mg, 65% yield).

[0232] Compound 140 1 H NMR (400 MHz, DMSO-d6) δ 8.96 (t, J = 5.9 Hz, 0.5H), 8.89 (t, J = 5.9 Hz, 0.5H), 8.74 (d, J = 4.8 Hz, 0.5H), 8.71 (d, J = 4.8 Hz, 0.5H), 7.97 (d, J = 1.9 Hz, 0.5H), 7.96 (d, J = 1.9 Hz, 0.5H), 7.77 (d, J = 1.9 Hz, 0.5H), 7.76 (d, J = 1.9 Hz, 0.5H), 7.57 (s, 1H), 7.28 (d, J = 4.8 Hz, 0.5H), 7.25 (d, J = 4.8 Hz, 0.5H), 6.36 - 6.02 (m, 1H), 4.87 - 4.75 (m, 2H), 3.94 (q, J = 9.0 Hz, 1H), 3.67 - 3.53 (m, 2H), 3.33 - 3.23 (m, 2H), 3.13 - 3.02 (m, 0.5H), 2.85 - 2.72 (m, 0.5H), 2.69 - 2.56 (m, 3H), 1.16 (d, J = 2.7 Hz, 3H), 1.03 (s, 1.5H), 1.00 (s, 1.5H).. HRMS (ESI): m / z exact mass calculated for C 28 H 24 Cl2F2N4O4S [M+H] + 621.0942, found 621.0931.

[0233] Example 61

[0234] Preparation of 3-((7-(3,5-dichloro-2-(3-(((2,2-difluoroethylamino)methyl)azetidine-1- carbonyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane- 2,4-dione (Compound 136).

[0235] Referring to Example 58, except that 2,2,2-trifluoroethyltrifluoromethanesulfonate was replaced with 2,2-difluoroethyltrifluoromethanesulfonate, the reaction yielded compound 136 as a white solid (37 mg, 55% yield). Compound 136... 1 H NMR (400MHz, DMSO-d6) δ8.73(d,J=4.9Hz,0.5H),8.72(d,J=4.9Hz,0.5H),7.96(t,J=1.8Hz,1H),7.79(d,J=2.0Hz,0.5H),7.76( d,J=2.0Hz,0.5H),7.57(d,J=2.3Hz,1H),7.27(d,J=4.9Hz,0.5H),7.25(d,J=4.9Hz,0.5H),6.08–5.68(m,1H),4.81(d,J=2.7Hz ,2H),3.93–3.79(m,1H),3.67–3.42(m,2H),3.22–3.16(m,0.5H),3.12–3.02(m,0.5H),2.88–2.75(m,1H),2.73–2.63(m,2H),2. 60(d,J=1.4Hz,2H),2.47–2.36(m,1H),2.24–1.92(m,2H),1.16(d,J=4.4Hz,3H),1.03(s,1.5H),1.00(s,1.5H)..HRMS(ESI):m / z exact mass calculated for C 28 H 26 Cl2F2N4O3S[M+H] + 607.1149, found 607.1137.

[0236] Example 62

[0237] Preparation of 3-((7-(2-(4-(aminomethyl)piperidin-1-carbonyl)-5-chloro-3-(trifluoromethyl)phenyl)thiophene[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (compound 122).

[0238] Referring to Example 17, the only difference being that 4-chloro-2-methylbenzoic acid was replaced with 4-chloro-2-trifluoromethylbenzoic acid to prepare a white solid (8 mg, 7% yield in four steps) of compound 122. Compound 122... 1H NMR (400 MHz, DMSO-d6) δ 8.73 (d, J = 4.9 Hz, 0.5H), 8.71 (d, J = 4.8 Hz, 0.5H), 8.19 - 8.08 (m, 2H), 7.82 (s, 2H), 7.57 (d, J = 5.6 Hz, 1H), 7.29 (d, J = 4.8 Hz, 0.5H), 7.21 (d, J = 4.8 Hz, 0.5H), 4.80 (d, J = 2.4 Hz, 2H), 4.15 (m, 1H), 3.22 - 3.15 (m, 0.5H), 3.04 - 2.97 (m, 0.5H), 2.80 - 2.66 (m, 1H), 2.64 - 2.60 (m, 2H), 2.47 - 2.37 (m, 1H), 2.34 - 2.24 (m, 1H), 2.15 - 2.05 (m, 0.5H), 1.97 - 1.88 (m, 0.5H), 1.64 - 1.23 (m, 4H), 1.18 (d, J = 3.2 Hz, 3H), 1.09 (s, 2H), 1.02 (s, 1H), 0.98 - 0.83 (m, 1H). HRMS (ESI): m / z exact mass calculated for C 29 H 28 ClF3N4O3S[M+H] + 605.1601, found 605.1584.

[0239] Example 63

[0240] 3-((7-(2-(3-(Aminomethyl)azetidine-l-carbonyl)-5-chloro-3- (trifluoromethyl)phenyl)thiophen-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 124).

[0241] Following the procedure of Example 18, except substituting 4-chloro-2- trifluoromethylbenzoic acid for 4-chloro-2-methylbenzoic acid, Compound 124 was prepared as a white solid (19 mg, 13% over four steps). Compound 124 1H NMR (400 MHz, DMSO-d6) δ 8.75 (m, 1H), 8.19 - 8.10 (m, 2H), 7.59 (d, J = 4.5 Hz, 1H), 7.37 - 6.76 (m, 3H), 4.81 (s, 2H), 3.89 - 3.70 (m, 1H), 3.66 - 3.58 (m, 0.5H), 3.53 - 3.47 (m, 0.5H), 3.46 - 3.40 (m, 1H), 3.22 - 3.14 (m, 1H), 2.82 - 2.70 (m, 1H), 2.60 (d, J = 3.3 Hz, 2H), 2.41 - 2.01 (m, 2H), 1.17 (d, J = 2.6 Hz, 3H), 1.05 - 0.97 (m, 3H). HRMS (ESI): m / z exact mass calculated for C 27 H 24 Cl2F3N4O3S [M+H] + 577.1288, found 577.1273.

[0242] Example 64

[0243] Preparation of 3-((7-(2-(4-(aminomethyl)piperidine-l-carbonyl)-3-chloro-5- (trifluoromethyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 127).

[0244] Following the procedure of Example 17, except substituting 2-chloro-4- trifluoromethylbenzoic acid for 4-chloro-2-methylbenzoic acid, Compound 127 was prepared as a white solid (22 mg, 5.4% over four steps). Compound 127 1H NMR (400 MHz, DMSO-d6) δ 8.75 (d, J = 4.8 Hz, 0.5H), 8.71 (d, J = 4.8 Hz, 0.5H), 8.20 (d, J = 1.6 Hz, 1H), 8.06 (d, J = 1.7 Hz, 0.5H), 8.00 (d, J = 1.7 Hz, 0.5H), 7.81 (s, 2H), 7.56 (d, J = 6.1 Hz, 1H), 7.29 (d, J = 4.8 Hz, 0.5H), 7.21 (d, J = 4.8 Hz, 0.5H), 4.80 (d, J = 2.9 Hz, 2H), 4.23 - 4.16 (m, 1H), 3.24 - 2.87 (m, 2H), 2.70 - 2.57 (m, 4H), 2.45 - 2.07 (m, 2H), 1.77 - 1.42 (m, 3H), 1.38 - 1.24 (m, 1H), 1.18 (d, J = 1.7 Hz, 3H), 1.07 (s, 1.5H), 1.03 (d, J = 5.9 Hz, 1.5H). HRMS (ESI): m / z exact mass calculated for C 29 H 28 ClF3N4O3S [M+H] + 605.1601, found 605.1585.

[0245] Example 65

[0246] 3-((7-(2-(3-(Aminomethyl)azetidine-l-carbonyl)-3-chloro-5- (trifluoromethyl)phenyl)thiophene[3,2-b]pyridine-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 126) was prepared according to the procedure described in Reference Example 18, with the exception that 4-chloro-2- methylbenzoic acid was replaced by 2-chloro-4-trifluoromethylbenzoic acid. Compound 126 was obtained as a light yellow solid (33 mg, 9% for four steps). Compound 126

[0247] Reference Example 18, with the exception that 4-chloro-2-methylbenzoic acid was replaced by 2-chloro-4-trifluoromethylbenzoic acid. Compound 126 was obtained as a light yellow solid (33 mg, 9% for four steps). Compound 126 1H NMR (400 MHz, DMSO-d6) δ 8.76 (d, J = 4.8 Hz, 0.5H), 8.74 (d, J = 4.8 Hz, 0.5H) 8.24 (d, J = 1.7 Hz, 0.5H), 8.22 (d, J = 1.7 Hz, 0.5H), 8.02 (s, 1H), 7.91 - 7.24 (m, 3H), 7.38 (d, J = 4.8 Hz, 0.5H), 7.30 (d, J = 4.8 Hz, 0.5H), 4.80 (s, 2H), 4.03 - 3.94 (m, 1H), 3.80 - 3.60 (m, 2H), 3.58 - 3.41 (m, 2H), 2.96 - 2.89 (m, 1H), 2.76 - 2.65 (m, 1H), 2.60 (s, 2H), 1.17 (s, 3H), 1.02 (d, J = 6.5 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C 27 H 24 ClF3N4O3S [M+H] + 577.1288, found 577.1274.

[0248] Example 66

[0249] Preparation of N-((1-(2-chloro-6-(2-((6,6-dimethyl-2,4-dioxo-3- azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2-b]pyridin-7-yl)-4- trifluoromethylbenzoyl)azetidin-3-yl)methyl)-2,2,2-trifluoroacetamide (Compound 130).

[0250] Referring to Example 59, the only difference is that 2,4-dichloromethylbenzoic acid is replaced by 2-chloro-4-trifluoromethylbenzoic acid to react to prepare light pink solid of Compound 130 (30 mg, yield 43%). The data of Compound 130 are as follows: 1H NMR (400 MHz, DMSO-d6) δ 9.56 (t, J = 5.8 Hz, 0.5H), 9.45 (d, J = 6.0 Hz, 0.5H),, 8.78 - 8.76 (d, J = 4.8 Hz, 0.5H), 8.73 (d, J = 4.8 Hz, 0.5H), 8.25 (d, J = 1.7 Hz, 0.5H), 8.22 (d, J = 1.7 Hz, 0.5H), 8.03 (s, 1H), 7.58 (s, 1H), 7.32 (d, J = 4.8 Hz, 0.5H), 7.29 (d, J = 4.8 Hz, 0.5H), 4.80 (d, J = 3.5 Hz, 2H), 4.03 - 3.94 (m, 1H), 3.72 - 3.57 (m, 2H), 3.43 - 3.32 (m, 2H), 3.20 - 3.08 (m, 0.5H), 2.89 (s, 0.5H), 2.75 - 2.61 (m, 1H), 2.58 (d, J = 2.3 Hz, 2H), 1.16 (d, J = 3.7 Hz, 3H), 1.03 (s, 1H), 0.99 (s, 2H). HRMS (ESI): m / z exact mass calculated for C 29 H 23 ClF6N4O4S[M+H] + 673.1111, found 673.1096.

[0251] Example 67

[0252] 3-((7-(3-chloro-2-(3-(((2,2,2-trifluoroethyl)amino)methyl)azetidine-l- carbonyl)-5-(trifluoromethyl)phenyl)thiophen-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 131).

[0253] Following the procedure of Example 58, except substituting 2-chloro-4- trifluoromethylbenzoic acid for 2,4-dichlorobenzoic acid, Compound 131 was prepared as a white solid (49 mg, 59% yield). Compound 131 1H NMR (400 MHz, DMSO-d6) δ 8.75 (d, J = 2.0 Hz, 0.5H), 8.74 (d, J = 2.0 Hz, 0.5H), 8.23 (d, J = 1.7 Hz, 0.5H), 8.22 (d, J = 1.7 Hz, 0.5H), 8.05 (d, J = 1.7 Hz, 0.5H), 8.02 (d, J = 1.7 Hz, 0.5H), 7.58 (s, 1H), 7.30 (t, J = 5.2 Hz, 1H), 4.81 (d, J = 4.0 Hz, 2H), 3.97 - 3.83 (m, 1H), 3.70 - 3.46 (m, 2H), 3.29 - 3.13 (m, 2H), 3.10 - 2.96 (m, 1H), 2.75 - 2.69 (m, 1H), 2.58 (s, 2H), 2.47 - 2.28 (m, 1H), 2.26 - 2.07 (m, 1H), 1.16 (d, J = 5.0 Hz, 3H), 1.02 (s, 1.5H), 0.99 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 29 H 25 ClF6N4O3S[M+H] + 659.1318, found 659.1304.

[0254] Example 68

[0255] 3-((7-(3-chloro-2-(5-oxa-2,8-diazaspiro[3.5]nonane-2-carbonyl)-5- (trifluoromethyl)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 132).

[0256] Referring to Example 1, the only difference is that 2,4-dichloromethylbenzoic acid is replaced by 2-chloro-4-trifluoromethylbenzoic acid, and tert-butyl 1,4-diazepane-1- carboxylate is replaced by tert-butyl 5-oxo-2,8-diazaspiro[3.5]nonane-8-carboxylate, the reaction to prepare Compound 132 white solid (28 mg, 10% yield for three steps) is prepared. Compound 132 1H NMR (400 MHz, DMSO-d6) δ 8.74 (t, J = 4.6 Hz, 1H), 8.24 (d, J = 1.6 Hz, 1H), 8.05 (d, J = 2.7 Hz, 1H), 7.58 (d, J = 1.7 Hz, 1H), 7.28 (d, J = 1.4 Hz, 1H), 4.80 (s, 2H), 3.85 (d, J = 10.7 Hz, 0.5H), 3.73 - 3.64 (m, 1.5H), 3.52 - 3.37 (m, 4H), 3.28 - 3.20 (m, 1H), 2.81 - 2.68 (m, 1H), 2.62 - 2.55 (m, 3H), 2.20 - 2.01 (m, 1H), 1.17 (d, J = 2.8 Hz, 3H), 1.03 (d, J = 6.8 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C 29 H 26 ClF3N4O4S [M+H] + 619.1394, found 619.1378.

[0257] Example 69

[0258] Preparation of 3-((7-(2-(4-(aminomethyl)piperidine-l-carbonyl)-5-chloro-3- (methyl-d3)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 129).

[0259] (1) Tetramethylethylenediamine (479 μL, 3.19 mmol) was placed in a dry three-necked flask, purged with argon 3 times, and anhydrous tetrahydrofuran solution (6 mL) was injected, and cooled to -78 °C. Secondary butyllithium (1.3 N, 2.65 mL) was slowly added. After stirring for 20 min, p-chlorobenzoic acid (200 mg, 1.28 mmol) was dissolved in anhydrous tetrahydrofuran solution (4 mL) and slowly injected into the system, and the system gradually changed from a yellow clear solution to an orange yellow suspension. After stirring for 2 h at -78 °C, deuterated iodomethane (119 μL, 1.92 mmol) was added, and stirred for 1 h. After the reaction was completed by TLC (petroleum ether: ethyl acetate = 1:4, with a drop of acetic acid), saturated ammonium chloride (5 mL) was added to quench the reaction. After being raised to room temperature, the phases were separated, the aqueous phase was washed with ethyl acetate (10 mL x 2), and the organic phase was discarded. Hydrochloric acid solution (1 N, 10 mL) was added to the aqueous phase to adjust the pH to 1-2, and a large amount of white solid was precipitated, which was filtered and dried to obtain the white solid of intermediate El (200 mg, yield 91%). The intermediate G1 was prepared according to the method described in WO2018123398. 1H NMR (400 MHz, DMSO) δ 12.96 (s, 1H), 7.83 (d, J = 8.4 Hz, 1H), 7.42 (d, J = 2.2 Hz, 1H), 7.35 (dd, J = 8.4, 2.3 Hz, 1H). HRMS (ESI): m / z exact mass calculated for C8H4D3ClO2 [M-H] - 172.0250, found 172.0246.

[0260] (2) Refer to Example 17, except that 4-chloro-2-methylbenzoic acid is replaced by intermediate Gl reaction to prepare compound 129 as a white solid (43 mg, 7% yield over four steps). Compound 129 has 1 H NMR (400 MHz, DMSO-d6) δ 8.69 (d, J = 4.8 Hz, 0.5H), 8.66 (d, J = 4.8 Hz, 0.5H), 7.84 (s, 2H), 7.62 - 7.51 (m, 3H), 7.24 (d, J = 4.8 Hz, 0.5H), 7.15 (d, J = 4.8 Hz, 0.5H), 4.79 (s, 2H), 4.29 - 4.18 (m, 1H), 3.42 - 3.35 (m, 1H), 3.20 - 3.12 (m, 0.5H), 3.06 - 2.99 (m, 0.5H), 2.91 - 2.72 (m, 1H), 2.67 - 2.57 (m, 3H), 2.56 - 2.52 (m, 0.5H), 2.37 - 2.12 (m, 2H), 2.03 - 1.94 (m, 0.5H), 1.71 - 1.31 (m, 3H), 1.18 (d, J = 2.4 Hz, 3H), 1.07 (s, 1.5H), 1.01 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 29 H 28 D3ClN4O3S [M+H] + 554.2072, found 554.2056.

[0261] Example 70

[0262] Preparation of 3-((7-(2-(3-(aminomethyl)azetidine-l-carbonyl)-5-chloro-3- (methyl-d3)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 137).

[0263] Referring to Example 18, the only difference being that 2,4-dichloromethylbenzoic acid was replaced with intermediate G1 in the reaction, yielding a white solid (20 mg, 12% yield in three steps) of compound 137. Compound 137... 1 H NMR(400MHz,DMSO-d6)δ8.74–8.69(m,1H),7.63–7.49(m,3H),7.29–6.97(m,3H),4.86– 4.75(m,2H),3.97–3.88(m,0.5H),3.84–3.75(m,0.5H),3.72–3.64(m,0.5H),3.63–3.54 (m,0.5H),3.51–3.42(m,1H),3.13–2.98(m,1H),2.94–2.83(m,1H),2.61(s,2H),2.46– 2.29(m,1.5H),2.23–2.08(m,0.5H),1.17(s,3H),1.01(d,J=8.8Hz,3H).HRMS(ESI):m / z exact mass calculated for C 27 H 24 D3ClN4O3S[M+H] + 526.1759, found 526.1744.

[0264] Example 71

[0265] Preparation of N-((1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexane-3-yl)methyl)thiophene[3,2-b]pyridin-7-yl)-6-(methyl-d3)benzoyl)azacyclobutane-3-yl)methyl)-2,2,2-trifluoroacetamide (compound 138).

[0266] Referring to Example 59, the only difference being that 2,4-dichloromethylbenzoic acid was replaced with intermediate G1 in the reaction, yielding compound 138 as a white solid (42 mg, 59% yield). Compound 138... 1H NMR (400 MHz, DMSO-d6) δ 9.52 (t, J = 5.9 Hz, 0.5H), 9.43 (t, J = 5.9 Hz, 0.5H), 8.72 (d, J = 4.7 Hz, 0.5H), 8.69 (d, J = 4.7 Hz, 0.5H), 7.66 - 7.50 (m, 3H), 7.24 (d, J = 4.7 Hz, 0.5H), 7.21 (d, J = 4.7 Hz, 0.5H), 4.87 - 4.74 (m, 2H), 3.92 (t, J = 8.3 Hz, 0.5H), 3.82 (t, J = 8.3 Hz, 0.5H), 3.63 - 3.56 (m, 1H), 3.41 - 3.35 (m, 1H), 3.31 - 3.22 (m, 1H), 3.11 - 3.00 (m, 0.5H), 2.99 - 2.88 (m, 0.5H), 2.59 (s, 3H), 2.48 - 2.38 (m, 1H), 1.16 (d, J = 6.2 Hz, 3H), 1.02 (s, 1.5H), 0.96 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 29 H 23 D3ClF3N4O4S[M+H] + 622.1582, found 622.1571.

[0267] Example 72

[0268] Preparation of N-((1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3- azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2-b]pyridin-7-yl)-6- (methyl-d3)benzoyl)azetidin-3-yl)methyl)-2,2-difluoroacetamide (Compound 139).

[0269] Reference to Example 60, except that 2,4-dichloromethylbenzoic acid was replaced by intermediate Gl reaction, white solid of Compound 139 was prepared (46 mg, yield 67%). The purity of Compound 139 was 99.0% (UV 210 nm), and the mass was 622.1571 Da. 1H NMR (400 MHz, DMSO-d6) 8.90 (t, J = 6.0 Hz, 0.5H), 8.81 (t, J = 6.0 Hz, 0.5H), 8.72 (d, J = 4.8 Hz, 0.5H), 8.69 (d, J = 4.8 Hz, 0.5H), 7.61 (d, J = 2.0 Hz, 0.5H), 7.58 (d, J = 2.0 Hz, 0.5H), 7.55 (d, J = 2.2 Hz, 1H), 7.53 (t, J = 2.0 Hz, 1H), 7.25 (d, J = 4.8 Hz, 0.5H), 7.20 (d, J = 4.8 Hz, 0.5H) 6.35 - 5.99 (m, 1H), 4.87 - 4.74 (m, 2H), 3.94 - 3.85 (m, 0.5H), 3.80 m, 0.5H), 3.64 - 3.51 (m, 1H), 3.41 - 3.35 (m, 1H), 3.32 - 3.22 (m, 2H), 3.09 - 2.98 (m, 0.5H), 2.94 - 2.81 (m, 0.5H), 2.59 (s, 2H), 2.52 (s, 0.5H), 2.38 (s, 0.5H), 1.16 (d, J = 4.9 Hz, 3H), 1.01 (s, 1.5H), 0.96 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 29 H 24 D3ClF2N4O4S[M+H] + 604.1676, found 604.1658.

[0270] Example 73

[0271] 3-((7-(5-chloro-3-(methyl-d3)-2-(3-(((2,2,2-trifluoroethyl)amino)methyl)azetidine-1- carbonyl)phenyl)thiophen-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 141).

[0272] Reference to Example 58, except that 2,4-dichloromethylbenzoic acid was replaced by intermediate Gl reaction, to produce Compound 141 as a white solid (15 mg, yield 22%). Compound 141 1H NMR (400 MHz, DMSO-d6) δ 8.71 (d, J = 3.0 Hz, 0.5H), 8.69 (d, J = 3.0 Hz, 0.5H), 7.63 - 7.51 (m, 3H), 7.23 (d, J = 4.8 Hz, 0.5H), 7.21 (d, J = 4.8 Hz, 0.5H), 4.88 - 4.74 (m, 2H), 3.91 - 3.82 (m, 0.5H), 3.75 - 3.66 (m, 0.5H), 3.57 - 3.50 (m, 1H), 3.31 - 3.22 (m, 1H), 3.21 - 3.08 (m, 1.5H), 3.04 - 2.86 (m, 1H), 2.85 - 2.78 (m, 0.5H), 2.65 (d, J = 7.4 Hz, 1H), 2.59 (d, J = 1.6 Hz, 2H), 2.42 - 2.14 (m, 2H), 2.00 - 1.81 (m, 1H), 1.15 (d, J = 7.9 Hz, 3H), 1.01 (s, 1.5H), 0.95 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 29 H 25 D3ClF3N4O3S[M+H] + 608.1789, found 608.1783.

[0273] Example 74

[0274] 3-((7-(5-chloro-2-(3-(((2,2-difluoroethylamino)methyl)azetidine-l- carbonyl)-3-(methyl-d3)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 142).

[0275] Refer to Example 61, the only difference is that 2,4-dichloromethylbenzoic acid is replaced by intermediate Gl reaction to prepare white solid of compound 142 (33 mg, yield 49%). The data of compound 142 1H NMR (400 MHz, DMSO-d6) δ 8.72 (d, J = 4.8 Hz, 0.5H), 8.70 (d, J = 4.8 Hz, 0.5H), 7.63 - 7.51 (m, 3H), 7.24 (d, J = 4.8 Hz, 0.5H), 7.21 (d, J = 4.8 Hz, 0.5H), 6.08 - 5.64 (m, 1H), 4.88 - 4.74 (m, 2H), 3.91 - 3.82 (m, 0.5H), 3.74 - 3.65 (m, 0.5H), 3.54 (d, J = 6.7 Hz, 1H), 3.32 - 3.23 (m, 1H), 3.15 - 3.06 (m, 0.5H), 2.86 - 2.72 (m, 1.5H), 2.68 - 2.53 (m, 4.5H), 2.40 - 2.20 (m, 1.5H), 1.83 (s, 1H), 1.15 (d, J = 7.0 Hz, 3H), 1.01 (s, 1.5H), 0.96 (s, 1.5H).. HRMS (ESI): m / z exact mass calculated for C 29 H 26 D3ClF2N4O3S[M+H] + 590.1884, found 590.1870.

[0276] Example 75

[0277] 3-((7-(5-chloro-2-(3-hydroxyazetidine-l-carbonyl)-3-(methyl-d3)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 143) was prepared according to the procedure described in Reference Example 1, with the exception that 2,4-dichloromethylbenzoic acid was replaced by the reaction of intermediate Gl, and tert-butyl 1,4-diazepane-l-carboxylate was replaced by 3-hydroxyazetidine hydrochloride (no need to remove Boc after coupling), to give Compound 143 as a white solid (30 mg, 18% yield over two steps). Compound 143

[0278] 3-((7-(5-chloro-2-(3-hydroxyazetidine-l-carbonyl)-3-(methyl-d3)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 143) was prepared according to the procedure described in Reference Example 1, with the exception that 2,4-dichloromethylbenzoic acid was replaced by the reaction of intermediate Gl, and tert-butyl 1,4-diazepane-l-carboxylate was replaced by 3-hydroxyazetidine hydrochloride (no need to remove Boc after coupling), to give Compound 143 as a white solid (30 mg, 18% yield over two steps). Compound 143 1H NMR (400 MHz, DMSO-d6) δ 8.69 (t, J = 4.5 Hz, 1H), 7.58 (d, J = 2.1 Hz, 0.5H), 7.57 (d, J = 2.1 Hz, 0.5H), 7.55 (d, J = 2.7 Hz, 1H), 7.52 (t, J = 2.0 Hz, 1H), 7.22 (d, J = 4.8 Hz, 0.5H), 7.20 (d, J = 4.8 Hz, 0.5H), 5.73 (d, J = 6.3 Hz, 0.5H), 5.56 (d, J = 5.9 Hz, 0.5H), 4.80 (d, J = 3.7 Hz, 2H), 4.38 - 4.32 (m, 0.5H), 4.11 - 4.02 (m, 0.5H), 4.00 - 3.96 (m, 0.5H), 3.80 - 3.69 (m, 1H), 3.63 - 3.55 (m, 0.5H), 3.54 - 3.47 (m, 0.5H), 3.47 - 3.39 (m, 0.5H), 3.28 - 3.20 (m, 0.5H), 3.00 - 2.96 (m, 0.5H), 2.59 (d, J = 1.4 Hz, 2H), 1.16 (s, 3H), 0.98 (d, J = 3.8 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C 26 H 21 D3ClN3O4S[M+H] + 513.1443, found 513.1435.

[0279] Example 76

[0280] Preparation of 3-((7-(5-chloro-2-(4-hydroxypiperidine-l-carbonyl)-3- (methyl-d3)phenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 144).

[0281] Following Example 1, except that 2,4-dichloromethylbenzoic acid was replaced by the reaction of intermediate Gl and tert-butyl 1,4-diazepane-l-carboxylate was replaced by 4-hydroxypiperidine hydrochloride (no need to remove Boc after coupling) to give Compound 144 as a white solid (29 mg, 16% yield over two steps). The purity of Compound 144 was 99.0% (UV). 1H NMR (400 MHz, DMSO-d6) 8.68 (d, J = 4.8 Hz, 0.5H), 8.66 (d, J = 4.8 Hz, 0.5H), 7.59 (d, J = 2.0 Hz, 0.5H), 7.58 (d, J = 2.0 Hz, 0.5H), 7.56 - 7.53 (m, 2H), 7.23 (d, J = 4.8 Hz, 0.5H), 7.20 (d, J = 4.8 Hz, 0.5H), 4.80 (d, J = 2.0 Hz, 2H), 4.61 (s, 1H), 3.73 - 3.61 (m, 1H), 3.52 - 3.41 (m, 1H), 3.16 - 2.92 (m, 2H), 2.87 - 2.79 (m, 1H), 2.59 (s, 2H), 2.49 (s, 1H), 1.59 - 1.48 (m, 1H), 1.27 - 1.21 (m, 2H), 1.17 (s, 3H), 1.01 (d, J = 4.9 Hz, 3H), 0.82 - 0.67 (m, 1H). HRMS (ESI): m / z exact mass calculated for C 28 H 25 D3ClN3O4S[M+H] + 541.1756, found 541.1746.

[0282] Example 77

[0283] 3-((7-(5-chloro-3-(methyl-d3)-2-(1-oxo-2,9-diazaspiro[5.5]undecan-9-carbonyl)phenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 145).

[0284] Following Example 1, except that 2,4-dichloromethylbenzoic acid was replaced by the reaction of intermediate Gl and tert-butyl 1,4-diazepane-1-carboxylate was replaced by 2,9-diazaspiro[5.5]undecan-1-one hydrochloride reaction (no need to remove Boc after coupling), white solid of Compound 145 was produced (27 mg, 10% yield for two steps). The purity of Compound 145 was 99.0% by HPLC. 1H NMR (400 MHz, DMSO-d6) δ 8.68 (d, J = 4.8 Hz, 0.5H), 8.65 (d, J = 4.8 Hz, 0.5H), 7.61 - 7.50 (m, 3H), 7.28 - 7.17 (m, 2H), 4.83 - 4.74 (m, 2H), 3.64 - 3.54 (m, 1H), 3.31 - 3.15 (m, 2H), 3.05 - 2.94 (m, 2.5H), 2.93 - 2.84 (m, 0.5H), 2.64 - 2.54 (m, 3H), 1.76 - 1.65 (m, 1H), 1.62 - 1.46 (m, 2H), 1.45 - 1.31 (m, 2H), 1.19 - 1.12 (m, 4H), 1.05 - 0.93 (m, 4H). HRMS (ESI): m / z exact mass calculated for C 32 H 30 D3ClN4O4S[M+H] + 608.2178, found 608.2164.

[0285] Example 78

[0286] 1-(4-chloro-2-(2-((6,6-dimethyl-2,4-dioxo-3-azabicyclo[3.1.0]hexan-3-yl)methyl)thieno[3,2- b]pyridin-7-yl)-6-(methyl-d3)benzoyl)piperidine-4-carboxamide (Compound 146).

[0287] Following Example 1, except that 2,4-dichloromethylbenzoic acid was replaced by the reaction of intermediate G1 and tert-butyl 1,4-diazepane-1-carboxylate was replaced by piperidine-4-carboxamide reaction (no need to remove Boc after coupling) to afford Compound 146 as a white solid (30 mg, 10% yield over two steps). Compound 146 1H NMR (400 MHz, DMSO-d6) δ 8.67 (d, J = 3.5 Hz, 0.5H), 8.65 (d, J = 3.4 Hz, 0.5H), 7.60 (d, J = 2.1 Hz, 0.5H), 7.58 (d, J = 2.1 Hz, 0.5H), 7.56 - 7.50 (m, 2H), 7.24 (d, J = 4.8 Hz, 0.5H), 7.20 (s, 0.5H), 7.16 (d, J = 4.7 Hz, 0.5H), 7.03 (s, 0.5H), 6.78 (s, 0.5H), 6.64 (s, 0.5H), 4.79 (d, J = 6.8 Hz, 2H), 4.23 - 4.11 (m, 1H), 3.20 - 3.13 (m, 0.5H), 3.09 - 3.01 (m, 0.5H), 2.90 - 2.80 (m, 0.5H), 2.71 - 2.64 (m, 0.5H), 2.62 - 2.56 (m, 2H), 2.34 - 2.24 (m, 1H), 2.11 - 2.04 (m, 1H), 1.61 - 1.49 (m, 1H), 1.38 - 1.21 (m, 3H), 1.16 (d, J = 3.0 Hz, 3H), 1.01 (s, 3H). HRMS (ESI): m / z exact mass calculated for C 29 H 26 D3ClN4O4S[M+H] + 568.1864, found 568.1853.

[0288] Example 79

[0289] 3-((7-(2-(3-(2-aminopropan-2-yl)azetidine-l-carbonyl)-5-chloro-3-(methyl-d3)phenyl)thiophen-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 147) was prepared according to the procedures described in Reference Example 1, with the exception that 2,4-dichloromethylbenzoic acid was replaced by the reaction of intermediate Gl and tert-butyl 1,4-diazepane-l-carboxylate was replaced by tert-butyl N-(2-(azetidin-3-yl)propan-2-yl)carbamate hydrochloride reaction to give Compound 147 as a white solid (39 mg, 9% yield over three steps). Compound 147 was characterized by the following data.

[0290] Example 79

[0289] 3-((7-(2-(3-(2-aminopropan-2-yl)azetidine-l-carbonyl)-5-chloro-3-(methyl-d3)phenyl)thiophen-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 147) was prepared according to the procedures described in Reference Example 1, with the exception that 2,4-dichloromethylbenzoic acid was replaced by the reaction of intermediate Gl and tert-butyl 1,4-diazepane-l-carboxylate was replaced by tert-butyl N-(2-(azetidin-3-yl)propan-2-yl)carbamate hydrochloride reaction to give Compound 147 as a white solid (39 mg, 9% yield over three steps). Compound 147 was characterized by the following data. 1H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 4.8 Hz, 0.5H), 8.66 (d, J = 4.8 Hz, 0.5H), 7.60 - 7.58 (m, 1H), 7.55 (d, J = 3.1 Hz, 1H), 7.54 - 7.52 (m, 1H), 7.25 (d, J = 4.8 Hz, 0.5H), 7.22 (d, J = 4.8 Hz, 0.5H), 6.49 (s, 2H), 4.85 - 4.77 (m, 2H), 3.90 - 3.74 (m, 1H), 3.71 - 3.59 (m, 1H), 3.49 - 3.43 (m, 1H), 3.24 - 3.17 (m, 0.5H), 3.11 - 3.03 (m, 0.5H), 2.69 - 2.61 (m, 0.5H), 2.59 (d, J = 1.3 Hz, 2H), 2.45 - 2.36 (m, 0.5H), 1.18 (s, 2H), 1.15 (s, 1H), 1.12 - 0.95 (m, 6H), 0.65 (s, 1.5H), 0.39 (s, 1.5H). HRMS (ESI): m / z exact mass calculated for C 29 H 28 D3ClN4O3S[M+H] + 554.2072, found 554.2059.

[0291] Example 80

[0292] 3-((7-(2-(3-(Aminomethyl)azetidine-l-carbonyl)-3-bromo-5-chlorophenyl)thieno[3,2- b]pyridin-2-yl)methyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2,4-dione (Compound 128) was prepared according to the procedure described in Reference Example 70, except that 2,4-dichlorobenzoic acid was replaced by 2-chloro-4-bromobenzoic acid reaction to give Compound 128 as a white solid (25 mg, 9% yield over three steps). Compound 128

[0293] Reference Example 70, except that 2,4-dichlorobenzoic acid was replaced by 2-chloro-4-bromobenzoic acid reaction to give Compound 128 as a white solid (25 mg, 9% yield over three steps). Compound 128 1H NMR (400 MHz, DMSO-d6) δ 8.77 - 8.68 (m, 1H), 8.09 (d, J = 2.0 Hz, 0.5H), 8.07 (d, J = 2.0 Hz, 0.5H), 7.80 (d, J = 1.9 Hz, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.30 (d, J = 4.8 Hz, 0.5H), 7.25 (d, J = 4.8 Hz, 0.5H), 6.68 (d, J = 8.0 Hz, 2H), 4.80 (s, 2H), 3.96 - 3.84 (m, 1H), 3.69 - 3.42 (m, 3H), 3.22 - 3.00 (m, 1H), 2.88 (d, J = 7.4 Hz, 1H), 2.60 (d, J = 5.4 Hz, 2H), 2.45 - 2.35 (m, 1H), 1.17 (d, J = 1.5 Hz, 3H), 1.03 (d, J = 7.5 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C 26 H 24 BrClN4O3S [M+H] + 587.0519, found 587.0507.

[0294] Example 81

[0295] Preparation of 3-((7-(2-(3-(hydroxymethyl)azetidin-1-yl)-5-chloro-3- methylphenyl)thieno[3,2-b]pyridin-2-yl)methyl)-6,6-dimethyl-3- azabicyclo[3.1.0]hexane-2,4-dione (Compound 93).

[0296] Following Example 27, except replacing piperidin-4-methanol with 3- methoxyazetidine hydrochloride (no need to remove Boc after coupling), Compound 93 was prepared as a brown solid (198 mg, 39% yield over two steps). Compound 128 was prepared following the procedure described in Example 128, except replacing 3- methoxyazetidine hydrochloride with 3-(hydroxymethyl)azetidine hydrochloride (no need to remove Boc after coupling), Compound 128 was prepared as a white solid (65 mg, 39% yield over two steps). 1H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 3.8 Hz, 0.5H), 8.69 (d, J = 3.8 Hz, 0.5H), 7.60 - 7.56 (m, 1H), 7.56 (s, 1H), 7.54 (d, J = 2.1 Hz, 0.5H), 7.52 (d, J = 2.1 Hz, 0.5H), 7.26 (d, J = 4.8 Hz, 0.5H), 7.22 (d, J = 4.8 Hz, 0.5H), 4.81 (d, J = 4.8 Hz, 2H), 4.62 (s, 1H), 3.86 (t, J = 9.0 Hz, 0.5H), 3.70 (t, J = 8.5 Hz, 0.5H), 3.66 - 3.60 (m, 0.5H), 3.31 - 3.19 (m, 2H), 3.55 - 3.48 (m, 0.5H), 2.99 - 2.86 (m, 1H), 2.72 - 2.62 (m, 0.5H), 2.59 (d, J = 1.7 Hz, 2H), 2.46 - 2.38 (m, 0.5H), 2.33 (d, J = 2.8 Hz, 4H), 1.16 (d, J = 3.5 Hz, 3H), 1.01 - 0.96 (m, 3H). HRMS (ESI): m / z exact mass calculated for C 27 H 26 ClN3O4S [M+H] + 524.1411, found 524.1398.

[0297] Example 82

[0298] 3-((7-(5-chloro-2-(4-(hydroxy(methyl)amino)piperidin-1-yl)-3- methy lpheny l)thieno [3,2-b]pyr idine-2-y l)methy l)-6,6-dimethy l-3- azabicyclo[3.1 0]hexane-2,4-dione (Compound 99)

[0299] (1) N-tert-butoxycarbonyl-4-piperidinone (199 mg, 1.0 mmol) was dissolved in dichloromethane (3 mL), and hydrogen chloride in n-hexane (4 N, 2.5 mL) was slowly added at 0 °C. After the addition was completed, the reaction was allowed to proceed at room temperature. After the reaction was completed, the solvent was removed under reduced pressure, and intermediate H1 (crude product, 100%) was prepared.

[0300] (2) Refer to the synthesis of intermediate A7 in Example 1, except that tert-butyl 1,4-diazepane-1-carboxylate was replaced with intermediate H1, and yellow crystalline solid of intermediate H2 was prepared (174 mg, yield 46.2%). 1H NMR (400 MHz, CDC13) δ 7.69 (dd, J = 1.9, 0.7 Hz, 1H), 7.23 (dd, J = 1.9, 0.8 Hz, 1H), 4.41 - 4.30 (m, 1H), 3.93 - 3.79 (m, 1H), 3.64 - 3.53 (m, 1H), 3.51 - 3.37 (m, 1H), 2.74 - 2.64 (m, 2H), 2.62 - 2.52 (m, 1H), 2.43 - 2.34 (m, 1H), 2.32 (s, 3H).

[0301] (3) Hydroxylamine hydrochloride (32 mg, 0.460 mmol) was dissolved in a mixed solvent of acetonitrile / water (2:1) (2 mL), sodium acetate (57 mg, 0.712 mmol) and H2(174 mg, 0.462 mmol) were added, and the reaction was carried out at 60°C. After the reaction was completed, which was monitored by TLC (petroleum ether: ethyl acetate = 1:1), the solvent was removed by reduced pressure, and the reaction was quenched with potassium carbonate aqueous solution. The aqueous phase was extracted with dichloromethane (20 mL x 2), and the combined organic phase was washed with saturated sodium chloride aqueous solution (20 mL x 2), dried over anhydrous sodium sulfate, and the solvent was removed by reduced pressure to obtain the white foamy solid of intermediate H3 (167 mg, crude yield 85.9%).

[0302] (4) Refer to the synthesis of intermediate A8 in Example 1, except that intermediate A7 was replaced by intermediate H3, to obtain the light orange solid of compound 100 (49.5 mg, yield 21.1%). 1 H NMR (400 MHz, DMSO) δ 10.47 - 10.43 (m, 1H), 8.68 (d, J = 4.8 Hz, 1H), 7.61 (d, J = 2.1 Hz, 1H), 7.58 (d, J = 1.8 Hz, 1H), 7.53 (s, 1H), 7.24 (d, J = 4.8 Hz, 1H), 4.79 (dd, J = 5.1, 2.4 Hz, 2H), 3.57 - 3.46 (m, 1H), 3.30 - 3.21 (m, 1H), 3.14 - 3.01 (m, 1H), 2.90 - 2.75 (m, 1H), 2.58 (dd, J = 5.5, 1.4 Hz, 2H), 2.28 (s, 3H), 2.27 - 2.09 (m, 2H), 1.89 - 1.66 (m, 2H), 1.16 (d, J = 3.9 Hz, 3H), 1.00 (d, J = 1.7 Hz, 3H). HRMS (ESI): m / z exact mass calculated for C 27 H 26 ClN3O4S [M+H] + 551.1520, found 551.1503.

[0303] Compounds 148-150, 154, 160-184 of the present application can be synthesized by the similar methods described above. The mass spectrometry data of some compounds are shown in the following table:

[0304] Example 83

[0305] Test of inhibitory activity of compounds on USP7 enzyme in vitro

[0306] 1. Preparation

[0307] Prepare enzyme activity test buffer: prepare a buffer containing 50 mM HEPES, 0.5 mM EDTA, 100 mM NaCl, 1 mM TCEP, 0.1 mg / mL BSA with deionized water, and adjust the pH to 7.5 with sodium hydroxide solution.

[0308] Prepare test solution: weigh an appropriate amount of test compound, dissolve it with DMSO, and dilute it by 3 times successively to obtain 8 concentrations (the highest concentration is adjusted according to the IC 50 of each compound). Weigh an appropriate amount of positive compound, dissolve it with DMSO, and dilute it by 3 times successively to obtain 8 concentrations (the highest concentration is 250 μM). The above compound solution and DMSO (blank control) are diluted 62.5 times with enzyme activity test buffer to obtain the test solution.

[0309] Prepare USP7 protein solution: dilute the commercially purchased USP7 protein solution (0.7 ug / ul, Recombinant USP7 Protein, Active Motif, Catalog No: 31525, Lot No: 08215001) with enzyme activity test buffer to 20 pM to obtain the USP7 protein solution for testing.

[0310] Prepare Ub-Rho solution: dilute the commercially obtained Ub-Rho solution (2.490 mg / mL, Boston Biochem, U-555-050, Lot No: DBFH2322121) 500 times with enzyme activity test buffer to obtain the Ub-Rho solution for testing.

[0311] 2. Sample addition and incubation

[0312] To the experimental wells of 384-well plates (Thermo Scientific, Lot No: 1207363), add USP7 protein solution (10 μL) and different concentrations of test compound solution (5 μL), 4 replicates for each concentration, and incubate the resulting mixture at room temperature in the dark for 30 min. Then add Ub-Rho solution (5 μL) to 2 replicates of each concentration, and enzyme activity test buffer (5 μL) to the other 2 replicates. After addition, incubate the 384-well plates at 37°C in the dark for 2 h. The positive control uses the USP7 inhibitor Compound 4 (CP4) reported in the literature (Nat Chem Biol, 2018, 14(2), 118).

[0313] 3. Readings and calculations

[0314] Read the fluorescence data with a multifunctional plate reader at an excitation wavelength of 485 nm and an emission wavelength of 535 nm. Inhibition rate % = [(F 空白(Ub-Rho)- F 空白(缓冲液A) )-(F 化合物(Ub-Rho) -F 化合物(缓冲液A) )] / (F 空白(Ub-Rho)- F 空白(缓冲液 A) )×100%.

[0315] IC 50 The numerical values are calculated with GraphPad Prism software.

[0316] 4. Experimental results

[0317] The inhibitory activity of some compounds on USP7 is shown in Table 1 below.

[0318] Table 1, Inhibition of USP7 enzyme activity by compounds of the present application

[0319] The above test results show that the compounds provided by the present application have strong USP7 inhibitory activity, especially compound 18, which reaches sub-nanomolar level, while the USP7 inhibitory activity of the comparative compounds WDL-04-123 and TSD-01-108 (amide nitrogen not in the ring), WDL-06-183 and WDL-06-173 (pyridine amide) is poor. At the same time, it is unexpectedly found that the activity of compound 20 is improved by 4 times after the piperazine ring is expanded to a seven-membered ring (compound 1), and the activity is well maintained after the piperazine ring is expanded to an eight-membered ring (compound 2) or N-methylated (compound 104).

[0320] Example 84

[0321] hERG inhibitory activity test of the compound

[0322] 1. Main experimental steps:

[0323] The concentration of the test sample was selected as 0.3 μM, 1 μM, 3 μM, 10 μM, 30 μM, and the concentration of the positive control sample was selected as 0.001 μM, 0.01 μM, 0.1 μM, 1 μM.

[0324] After the HEK-293-hERG cells were subcultured to a suitable state, the cells were rinsed with PBS (or DPBS), digested and separated with a Tryple solution, resuspended in a culture medium and stored in a centrifuge tube, the supernatant was discarded after centrifugation, and the cells were resuspended in an extracellular solution for standby, and stored at 2-8°C. Before patch clamp recording, the cells were added dropwise to a culture dish to ensure that the cells had a certain density and were in a single isolated state.

[0325] The hERG current was recorded by whole-cell patch clamp technique. The cell suspension was added to a small culture dish and placed on an inverted microscope stage. After the cells adhered, they were perfused with an extracellular solution at a recommended flow rate of 1-2 mL / min. The glass microelectrode was two-step drawn by a microelectrode puller, and after being filled with the electrode internal solution, its water resistance value was 2-5 MΩ.

[0326] After establishing the whole-cell recording mode, the clamping potential was maintained at -80 mV. A depolarization voltage of +60 mV was given for 850 ms, and then repolarization to -50 mV was maintained for 1275 ms to induce hERG tail current. This set of pulse programs was repeated every 15 seconds throughout the experiment.

[0327] After the current was stable, the drug was administered by continuous perfusion from low concentration to high concentration. Starting from a low concentration, the perfusion was continued until the drug effect was stable, and then the perfusion of the next concentration was performed. In this experiment, the blocking effect of each concentration of the test sample and the positive control on the hERG tail current was tested (N≥2); the actual concentration can be adjusted according to the actual solubility and effect, and is not considered as a deviation from the scheme.

[0328] The definition of drug effect stability is that the current value change of the last 5 stimulation bars at each concentration administration stage is less than 10% of the mean value (when the current is greater than or equal to 200 pA) or less than 30% of the mean value (when the current is less than 200 pA), which is considered stable, and if not stable, the data at that concentration will not be taken.

[0329] Stimulation and signal acquisition were performed by PatchMaster software; the patch clamp amplifier amplified the signal.

[0330] Further data analysis and curve fitting were performed using FitMaster, EXCEL, Graphpad Prism or SPSS21.0, etc. The data were expressed as mean and standard deviation.

[0331] In data processing, the peak value of the tail current and its baseline are corrected when judging the blocking effect on hERG. The inhibition rate (IR) of the tail current is used to represent the effect of each compound at different concentrations. The SD of %IR of all cells at each concentration is ≤ 15, which is the acceptable standard (except for abnormal data). IR = 100% x (peak value of tail current before administration-peak value of tail current after administration) / peak value of tail current before administration.

[0332] IC 50 The numerical value is obtained by fitting the Hill equation (if applicable), and if the maximum inhibition rate of the test substance at all concentrations is less than 50%, the IC is not calculated 50 The numerical value.

[0333] 2、Experimental results

[0334] The inhibition activity of some compounds on hERG is shown in Table 2.

[0335] Table 2, Inhibition of compounds of the present application on hERG

[0336] The above test results show that the compounds provided by the present application have low inhibition activity on hERG (IC 50 ≥ 8 μM), especially compound 8 and compound 137 (IC 50 > 30 μM, which is the generally recognized low risk standard of cardiotoxic side effects), which is significantly lower than the hERG inhibition activity of the comparative compound CP41 (IC 50 = 4.2 μM, J. Med. Chem. 2020, 63, 10, 5398), indicating that the compounds provided by the present application have a lower risk of cardiotoxic side effects.

[0337] Example 85

[0338] Compound human liver microsomal stability test

[0339] 1、Main experimental steps

[0340] The compound was added to the incubation system containing human liver microsomes and NADPH regeneration system, respectively, and incubated. The reaction was terminated at the specified time 0, 10, 20, 30, 60 min (n = 3) and the sample was collected; an LC-MS / MS method was established to determine and calculate the area ratio of the compound and the internal standard compound, and the metabolic residual rate-time curve was drawn using the GraphPad Prism 8.0 drawing software; the in vitro metabolism related parameters were calculated by the statistical moment method using the pharmacokinetics professional software WinNonlin 7.0.

[0341] 2、Experimental results

[0342] The stability of the compounds in human liver microsomes is shown in Table 3.

[0343] Table 3, Test results of the stability of the compounds of the present application in human liver microsomes

[0344] The above test results show that the compounds 1, 8, 123, 129, 137, etc. provided by the present application have high stability in human liver microsomes (T 1 / 2 > 40 min), indicating that they can have good in vivo PD / PK properties and good drug properties.

[0345] Example 86

[0346] Preparation of tablets:

[0347] The compound 1 prepared in Example 1 (5 g), hydroxypropyl methyl cellulose E (150 g), starch (200 g), polyvinylpyrrolidone K30 (appropriate amount), and magnesium stearate (1 g) were mixed, granulated, and tableted.

[0348] In addition, according to the conventional preparation method of Pharmacopoeia 2015 edition, the compounds 1-17 can be endowed with different pharmaceutical excipients to form capsules, powders, granules, pills, injections, syrups, oral solutions, inhalants, ointments, suppositories, or patches, etc.

Claims

A benzamide compound of Formula (I) or a pharmaceutically acceptable salt, ester, stereoisomer, deuterated form, or solvate thereof: wherein: R 1 , R 2 are each independently selected from H, halogen, -CN, alkyne, -N3, -OR 4 , -OCOR 4 , -SR 4 , -NR 4 R 5 , -N(R 4 )COR 5 , -COOR 4 , -CON(R 4 )R 5 , -SF5, C 1-3 alkyl or deuterated alkyl or cycloalkyl or halogenated alkyl or halogenated cycloalkyl; Ring A is unsubstituted or contains a substituent R 3a , R 3b of a 4-8 membered nitrogen-containing non-aromatic heterocycle; R 3a , R 3b each independently is selected from H, halogen, =0, =NOH, =NOR 6 , -CN, -N3, -OR 6 , -OCOR 6 , -SR 6 , -NR 6 R 7 , -N(R 6 )COR 7 , -COOR 6 , -CON(R 6 )R 7 , -SO2R 6 , -SO2N(R 6 )R 7 , C 1-6 1-6 alkyl, C 2-6 2-6 alkenyl, C 2-6 2-6 alkynyl, 3-8 membered cycloalkyl or heterocycloalkyl, 5-10 membered aryl or heteroaryl, which alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted with halogen, -CN, -COOR 8 , -CON(R 8 )R 9 , -NO2, -OR 8 , -N(R 8 )R 9 , C 1-6 1-6 alkyl, C 2-6 2-6 alkenyl, C 2-6 2-6 alkynyl, C 3-8 3-8 cycloalkyl or C 3-8 3-8 membered heterocycloalkyl; or R 3a and R 3b , when attached to the same ring atom, together form a 4-7 membered ring; M is a carbon atom or a nitrogen atom to which the substituents Z are attached, when M is a carbon atom C the substituents Z are attached 3 , Z 4 , when M is a nitrogen atom N the substituents Z are attached 3 , Z 1 , Z 2 , Z 3 , Z 4 are each independently selected from H, halogenated or unhalogenated alkyl or cycloalkyl, Z 1-3 , Z 1 , Z 2 , Z 3 , Z 4 Any one of Z1, Z2, Z3, Z4, Z5, Z6, Z7, Z8, Z9, Z10, Z11, Z12, Z13, Z14, Z15, Z16, Z17, Z18, Z19, Z20, Z21, Z22, Z23, Z24, Z25, Z26, Z27, Z28, Z29, Z30, Z31, Z32, Z33, Z34, Z35, Z36, Z37, Z38, Z39, Z40, Z41, Z42, Z43, Z44, Z45, Z46, Z47, Z48, Z49, Z50, Z51, Z52, Z53, Z54, Z55, Z56, Z57, Z58, Z59, Z60, Z61, Z62, Z63, Z64, Z65, Z66, Z67, Z68, Z69, Z70, Z71, Z72, Z73, Z74, Z75, Z76, Z77, Z78, Z79, Z80, Z81, Z82, Z83, Z84, Z85, Z86 R 4 , R 5 , R 6 , R 7 , R 8 , R 9 each independently is selected from H, halogen, C 1-6 alkyl or deuterated alkyl or halogenated alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, 3- to 8-membered cycloalkyl or heterocycloalkyl, 5- to 10-membered aryl or heteroaryl. The benzamide compound or a pharmaceutically acceptable salt, ester, stereoisomer, deuteride, or solvate thereof according to Claim 1, characterized in that, The selected from the group consisting of: wherein m, p, q, y are each independently selected from 1, 2 or 3; R 3a , R 3b each independently taken from H, -OH, halogen, -NH2, -NHMe, -NMe2, -CH2OH, -CH2NH2, -CH2NHMe, -CH2NMe2, -COOH, -CONH2, -CH2NHCOCHMe2, -CH2NHCOOCMe3, -CHMeNH2, -CH2COOEt, -CH2NH2COOMe, -CH2NHCOOCHMe2, -CH2NHCOMe, -CH2NHPyr, -CH2NHCONH2, -CH2NHCONHEt, -CH2NHCOCF3, -CH2NHCH2CF3, -CH2NHCH2CHF2, -CH2NHCOCHF2, -CMe2NH2, -CONHMe, -CMe2NHMe, -CMe2NHCOCF3, CHMeNHCOCF3, R c , -CH2NHCOMe, -CH2NHPyr, -CH2NHCONH2, -CH2NHCONHEt, -CH2NHCOCF3, -CH2NHCH2CF3, -CH2NHCH2CHF2, -CH2NHCOCHF2, -CMe2NH2, -CONHMe, -CMe2NHMe, -CMe2NHCOCF3, CHMeNHCOCF3, R 3a , -CH2NHCOMe, -CH2NHPyr, -CH2NHCONH2, -CH2NHCONHEt, -CH2NHCOCF3, -CH2NHCH2CF3, -CH2NHCH2CHF2, -CH2NHCOCHF2, -CMe2NH2, -CONHMe, -CMe2NHMe, -CMe2NHCOCF3, CHMeNHCOCF3, R 3b are not simultaneously H; X is selected from -0-, -S-, -NH-, -N(Me)-, -N(Et)-, -N(Pr i ), -N(Pr c ), -N(CONH2)-, -C(=NOH)- or -C(=NOH)-. The benzamide compound or a pharmaceutically acceptable salt, ester, stereoisomer, deuteride, or solvate thereof according to claim 1, characterized in that, The selected from the group consisting of The benzamide compound or a pharmaceutically acceptable salt, ester, stereoisomer, deuteride or solvate thereof according to any one of claims 1 to 3, characterized in that, The compound is selected from any one of the following compounds: Use of a compound of any one of claims 1-4, or a pharmaceutically acceptable salt, ester, stereoisomer, deuterated analog, or solvate thereof, in the manufacture of a USP7 inhibitor. Use of a compound of any one of claims 1-4, or a pharmaceutically acceptable salt, ester, stereoisomer, deuterated analog, or solvate thereof, in the manufacture of a medicament for preventing or treating a USP7-mediated disease. The use according to claim 6, characterized in that The USP7-mediated disease is any one of a tumor, a viral infectious disease, an inflammatory disease, or an autoimmune disease. The use according to claim 7, characterized in that The tumor disease is bone cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, myeloproliferative disease, multiple myeloma, myelodysplastic syndrome, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hemangioma, granuloma, xanthoma, meningiosarcoma, glioma, neuroblastoma, astrocytoma, medulloblastoma, ependymoma, germinoma, glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, fibroma, sarcoma, esophageal cancer, stomach cancer, pancreatic cancer, large intestinal cancer, colon cancer, rectal cancer, kidney cancer, prostate cancer, lymphatic cancer, testicular cancer, interstitial cell carcinoma, lung cancer, liver cancer, skin cancer, basal cell carcinoma, or soft tissue tumor; the viral infectious disease is an infectious disease caused by an RNA virus or a DNA virus. A pharmaceutical composition for the preparation of a medicament for preventing or treating a USP7-mediated disease, characterized in that, The composition comprises a compound of any one of claims 1-4, or a pharmaceutically acceptable salt, ester, stereoisomer, deuterated analog, or solvate thereof, and a pharmaceutically acceptable carrier. The pharmaceutical composition according to claim 9, characterized in that, The pharmaceutical composition is a capsule, a powder, a tablet, a granule, a pill, an injection, a syrup, an oral solution, an inhalant, an ointment, a suppository, or a patch.

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