Oxygen-containing heterocyclic compound and use thereof

Abstract

Disclosed herein are an oxygen-containing heterocyclic compound and uses thereof. The present disclosure provides an oxygen-containing heterocyclic compound represented by formula I, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt or a crystal form thereof. The oxygen-containing heterocyclic compound is expected to selectively inhibit Smad3 activation. # imgabs0 #

Description

[0001] Cross - reference to related applications

[0002] This application claims priority to U.S. Provisional Application 63 / 370,516, filed on August 5, 2022, the disclosure of which is hereby incorporated by reference in its entirety. Field of the Invention

[0003] The present invention relates to oxygen - containing heterocyclic compounds and their uses. Background Art

[0004] Fibrosis is characterized by the over - production and accumulation of extracellular matrix proteins, which leads to a gradual loss of tissue function and ultimately organ failure. Chronic kidney disease is usually accompanied by renal interstitial fibrosis regardless of the primary insult. To prevent renal function decline, treatment strategies for chronic kidney disease not only need to eliminate etiological factors such as hyperglycemia, hypertension, and HIV infection, but also require anti - fibrosis treatment to restore normal renal structure and function. In addition to kidney fibrosis, many other organ - specific fibrosis disorders are also known, including liver, heart, and lung fibrosis.

[0005] Regardless of the original etiology of the disease, renal fibrosis is considered the final convergent pathway of progressive chronic kidney disease. Although much is known about the molecular mechanisms of renal fibrosis, there has been limited success in translating this knowledge into clinical applications. HIPK2 has been shown to be a multifunctional activator of the TGF - β / Smad3, NF - κB, and p53 pathways, and global knockout of HIPK2 in mice can reduce renal fibrosis in vivo. U.S. Patent 10,669,266 discloses small - molecule inhibitors of HIPK2 that specifically block the TGF - β / Smad3 pathway to reduce renal fibrosis without causing adverse systemic effects. However, the compounds disclosed in the '266 patent still have solubility and potency issues.

[0006] Transforming growth factor - β1 (TGF - β1) has been identified as the most important profibrotic factor in kidney disease. TGF - β1 binds to the type II TGF - β receptor, causing it to dimerize with the type I TGF - β receptor and resulting in the phosphorylation of Smad2 and Smad3. Phosphorylated Smad3 relocates to the nucleus, where it binds to Smad - binding elements in the promoter and activates the transcription of target genes, including profibrotic genes such as collagen I, fibronectin, and α - smooth muscle actin (α - SMA). Smad3 is known to be highly active in fibrotic kidneys, and knockout of Smad3 can reduce renal fibrosis in animal models of kidney disease. Thus, blocking the TGF - β1 / Smad3 pathway provides a treatment strategy for kidney fibrosis. Summary of the Invention

[0007] The technical problem to be solved by the present invention is the lack of effective Smad3 activation inhibitors in the prior art for clinical treatment. Therefore, the present invention provides oxygen-containing heterocyclic compounds that can selectively inhibit Smad3 activation and their uses.

[0008] The present disclosure solves the above technical problems through the following technical solutions.

[0009] The present disclosure provides an oxygen-containing heterocyclic compound represented by Formula I, its pharmaceutically acceptable salt, its deuterated derivative, its solvate, the solvate of the pharmaceutically acceptable salt, or its crystal form:

[0010]

[0011] Wherein:

[0012] Ar is C 6-20 aryl, C 1-1 aryl substituted with one or more R 6-20 "5-12 membered heteroaryl containing 1-4 heteroatoms selected from O, S, and N", or "5-12 membered heteroaryl containing 1-4 heteroatoms selected from O, S, and N" substituted with one or more R 1-2 ;

[0013] R 1-1 and R 1-2 are independently selected from halogen, hydroxy, and C 1-6 alkyl;

[0014] is a 5-6 membered heteroaryl containing 1-3 heteroatoms selected from O, S, and N;

[0015] R 1 and R 2 are independently selected from hydrogen, deuterium, hydroxy, C 1-6 alkyl, amino, and -OC 1-6 alkyl;

[0016] R 3 and R 4 are C 1-6 alkyl substituted with deuterium, or, R 3 and R 4 together with the atom to which they are attached form C 3-8 cycloalkyl, C 1-3 cycloalkyl substituted with R 3-8 "4 to 10 membered heterocycloalkyl containing 1-3 heteroatoms selected from O, S, and N", or "4 to 10 membered heterocycloalkyl containing 1-3 heteroatoms selected from O, S, and N" substituted with R 1-4 ;

[0017] R 1-3 and R1-4 independently selected from halogen, deuterium, hydroxyl, and C 1-6 alkyl.

[0018] In one embodiment, the oxygen-containing heterocyclic compound represented by Formula I has the structure of Formula II:

[0019]

[0020] wherein represents or and a mixture of;

[0021] n is 1, 2, 3, 4, or 5;

[0022] Ar and are as defined in Formula I.

[0023] In one embodiment, the oxygen-containing heterocyclic compound represented by Formula I has the structure of Formula III:

[0024]

[0025] wherein:

[0026] Ar and are as defined in Formula I;

[0027] represents or and a mixture of;

[0028] R 3 and R 4 are C 1-6 alkyl substituted with deuterium.

[0029] In one embodiment, Formula III has the following structure:

[0030]

[0031] wherein Ar and are as defined in Formula III.

[0032] In one embodiment, in Formula II or III',

[0033] Ar is a C 1-1 aryl substituted with one or more R 6-20 or a "5-12 membered heteroaryl containing 1-4 heteroatoms selected from O, S, and N" substituted with one or more R 1-2 ;

[0034] R1-1 and R 1-2 are independently selected from halogen and C 1-6 alkyl;

[0035] is a 5- or 6-membered heteroaryl containing 1 to 3 heteroatoms independently selected from O, S, and N;

[0036] represents

[0037] n is 2 or 3.

[0038] In one embodiment, in Formula II,

[0039] Ar is a C 1-1 aryl substituted with one or more R 6-20 or a "5- to 12-membered heteroaryl containing 1 to 4 heteroatoms independently selected from O, S, and N" substituted with one or more R 1-2 ;

[0040] R 1-1 and R 1-2 are independently selected from halogen and C 1-6 alkyl;

[0041] is a 5- or 6-membered heteroaryl containing 1 to 3 heteroatoms independently selected from O, S, and N;

[0042] represents

[0043] n is 2 or 3.

[0044] In one embodiment, in Formula II or III',

[0045] Ar is a C 1-1 aryl substituted with one or more R 6-20 ;

[0046] R 1-1 is halogen;

[0047] is a 5- or 6-membered heteroaryl containing 1 to 3 heteroatoms selected from O, S, and N;

[0048] represents

[0049] n is 2.

[0050] In one embodiment, n is 1, 2, or 3.

[0051] In one embodiment, Ar is a C1-1 Substituted C 6-20 aryl or a 5- to 12-membered heteroaryl containing one or more R 1-2 substituents and having 1-4 heteroatoms independently selected from O, S, and N.

[0052] In one embodiment, R 1-1 and R 1-2 are independently selected from halogen, and C 1-6 alkyl;

[0053] In one embodiment, is a 5-membered heteroaryl containing 3 heteroatoms selected from O and N.

[0054] In one embodiment, R 3 and R 4 together with the atoms to which they are attached form a 3- to 8-membered ring.

[0055] In one embodiment, when Ar is C 6-20 aryl, or C 1-1 aryl substituted with one or more R 6-20 substituents, the C 6-20 aryl and the C 1-1 aryl in the C 6-20 aryl substituted with one or more R 6-20 substituents are C 6-10 aryl; the C 6-10 aryl is phenyl or naphthyl;

[0056] In one embodiment, when Ar is a "5- to 12-membered heteroaryl containing 1-4 heteroatoms independently selected from O, S, and N" or a "5- to 12-membered heteroaryl containing 1-4 heteroatoms independently selected from O, S, and N" substituted with one or more R 1-2 substituents, the "5- to 12-membered heteroaryl containing 1-4 heteroatoms independently selected from O, S, and N" in the "5- to 12-membered heteroaryl containing 1-4 heteroatoms independently selected from O, S, and N" and the "5- to 12-membered heteroaryl containing 1-4 heteroatoms independently selected from O, S, and N" substituted with one or more R 1-2 substituents is a "5-membered heteroaryl containing 2 heteroatoms selected from N", such as pyrazolyl

[0057] In one embodiment, when is a 5- to 6-membered heteroaryl containing 1-3 heteroatoms independently selected from O and N, the is a 5-membered heteroaryl containing 3 heteroatoms independently selected from O and N, for example,

[0058] In one embodiment, when R 1-1 and R 1-2 are independently selected from halogen, the halogen is -F, -Cl, -Br or -I.

[0059] In one embodiment, when R 1-1 and R 1-2 are independently selected from C 1-6 alkyl, the C 1-6 alkyl is C 1-4 alkyl, or can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, or can be methyl.

[0060] In one embodiment, when R 3 and R 4 together with the atom to which they are attached form a C 3-8 cycloalkyl, the C 3-8 cycloalkyl is C3 cycloalkyl, C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl or C8 cycloalkyl.

[0061] In one embodiment, when R 3 and R 4 are deuterium-substituted C 1-6 alkyl, the C 1-6 alkyl is C 1-4 alkyl, or can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, or can be methyl.

[0062] In one embodiment, when R 3 and R 4 are deuterium-substituted C 1-6 alkyl, the deuterium-substituted C 1-6 alkyl is -CD3.

[0063] In one embodiment, Ar is

[0064] In one embodiment, is

[0065] In one embodiment, when is then, either side of

[0066] In one embodiment, is

[0067] In one embodiment, is

[0068] In one embodiment, the oxygen-containing heterocyclic compound of Formula I may have any of the following structures:

[0069]

[0070] The present disclosure also provides a pharmaceutical composition comprising Substance A and a pharmaceutically acceptable excipient, wherein Substance A is a therapeutically effective amount of the oxygen-containing heterocyclic compound of Formula I as described above, its pharmaceutically acceptable salt, its deuterated derivative, its solvate, the solvate of the pharmaceutically acceptable salt, or its crystal form.

[0071] In the present disclosure, the pharmaceutical composition can be in the form of an oral dosage form or in the form of a sterile injectable aqueous solution, and can be prepared according to any method for preparing pharmaceutical compositions known in the art.

[0072] The present disclosure also provides a method for inhibiting the interaction between homeodomain interacting protein kinase 2 and Smad3, the method comprising binding HIPK2 to Substance A, wherein Substance A is a therapeutically effective amount of the oxygen-containing heterocyclic compound of Formula I as described above, its pharmaceutically acceptable salt, its deuterated derivative, its solvate, the solvate of the pharmaceutically acceptable salt, or its crystal form.

[0073] The present disclosure also provides a method for inhibiting Smad3 activation, the method comprising contacting Smad3 with Substance A, wherein Substance A is a therapeutically effective amount of the oxygen-containing heterocyclic compound of Formula I as described above, its pharmaceutically acceptable salt, its deuterated derivative, its solvate, the solvate of the pharmaceutically acceptable salt, or its crystal form.

[0074] The present disclosure also provides a method for treating a fibrotic disease, comprising administering Substance A to a subject suffering from a fibrotic disease, wherein Substance A is a therapeutically effective amount of the oxygen-containing heterocyclic compound of Formula I as described above, its pharmaceutically acceptable salt, its deuterated derivative, its solvate, the solvate of the pharmaceutically acceptable salt, or its crystal form, and the disease is renal fibrosis, cardiac fibrosis, hepatic fibrosis, or pulmonary fibrosis.

[0075] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention prepared with relatively safe and pharmaceutically acceptable acids or bases. When the compounds disclosed herein contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of the compound with a sufficient amount of a pharmaceutically acceptable base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to: lithium salts, sodium salts, potassium salts, calcium salts, aluminum salts, magnesium salts, zinc salts, bismuth salts, ammonium salts, and diethanolamine salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of the compound with a sufficient amount of a pharmaceutically acceptable acid in a pure solution or a suitable inert solvent. Pharmaceutically acceptable acids include inorganic acids, including but not limited to: hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, and sulfuric acid. Pharmaceutically acceptable acids include organic acids, including but not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acid citrate, oleic acid, tannic acid, pantothenic acid, acid tartrate, ascorbic acid, gentisic acid, fumaric acid, gluconic acid, glucaric acid, formic acid, ethanesulfonic acid, pamoic acid (i.e., 4,4'-methylene-bis(3-hydroxy-2-naphthoic acid)), and amino acids (e.g., glutamic acid and arginine). When the compounds disclosed herein contain both relatively acidic functional groups and relatively basic functional groups, they can be converted into base addition salts or acid addition salts. For details, see Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66:1-19 (1977), or Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl and Camille G. Wermuth, ed., Wiley-VCH, 2002).

[0076] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0077] The term "alkyl" refers to straight-chain or branched-chain alkyl groups having a specific number of carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc.

[0078] The terms "cycloalkyl" and "carbocyclic ring" refer to saturated cyclic groups composed only of carbon atoms having a specific number of carbon atoms (e.g., C3-C6), which are monocyclic, bridged, or spirocyclic. Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0079] The term "heteroaryl" refers to a cyclic group having a specified number of ring atoms (e.g., 5 - 12 members), a specified number of heteroatoms (e.g., 1, 2, or 3), and specified heteroatom species (one or more of N, O, and S), which cyclic group is monocyclic or polycyclic and has at least one aromatic ring (according to Hückel's rule). The heteroaryl is connected to other fragments of the molecule through an aromatic ring or a non - aromatic ring. Heteroaryls include, but are not limited to, furyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, pyridyl, pyrimidinyl, and indolyl.

[0080] The terms "heterocyclic group", "heterocycle", or "heterocycloalkyl" refer to a cyclic group having a specific number of ring atoms (e.g., 3 - 8 members), a specific number of heteroatoms (e.g., 1, 2, or 3), and specific heteroatom species (one or more of N, O, and S), which cyclic group is monocyclic, bridged, or spirocyclic and each ring is saturated. Heterocycloalkyls include, but are not limited to, azetidinyl, pyrrolidinyl, tetrahydrofuryl, morpholinyl, piperidinyl, etc.

[0081] The term "hydroxy" refers to the - OH group.

[0082] The term "cyano" refers to the - CN group.

[0083] Experimental Section

[0084] Example 1: Synthetic Route of Compound 1

[0085]

[0086] Synthesis of Compound 1 - j

[0087] Hydroxylamine (21.13 g, 319.85 mmol) was added to a mixture of 4 - chlorobenzonitrile (11 g, 79.96 mmol) in EtOH (150 mL). The mixture was stirred at 90 °C for 2 hours. After the consumption of the starting material (determined by LCMS), the mixture was concentrated in vacuo and the product was dried by lyophilization to obtain Compound 1 - j (13 g, crude).

[0088] Synthesis of Compound 1 - i

[0089] To a round-bottom flask (100 mL) containing compound 1-j (700 mg, 4.10 mmol) was added 1,1-carbonyldiimidazole (CDI, 642.14 mg, 4.46 mmol) and 1,8-diazabicyclo[5.4.0]-7-undecene (DBU, 1.23 g, 4.87 mmol). Then 1,4-dioxane (10 mL) was added in turn, and the reaction mixture was stirred at 100 °C for 3 h. After cooling to ambient temperature, the mixture was diluted with water, adjusted to pH ~2 with 3M aqueous HCl, and extracted with ethyl acetate (30 mL * 2). The combined organic phases were washed with water, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by silica gel column chromatography (EA:PE = 0 - 15%) to obtain compound 1-i (400 mg, 50% yield).

[0090] Synthesis of compound 1-h

[0091] A mixture of compound 1-i (1.0 g, 5.09 mmol) and POCl3 (16.45 g, 105.90 mmol, 10 mL) was added to 1,8-diazabicyclo[5.4.0]-7-undecene (DBU, 1.53 g, 10.05 mmol, 1.5 mL) at 0 °C. The mixture was stirred at 100 °C for 12 h. The mixture was poured into ice water, and the pH was adjusted to 9 by adding saturated NaHCO3. The mixture was extracted with ethyl acetate (30 mL * 3). The combined organic phases were washed with water, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo, and purified by silica gel column chromatography (EA:PE = 0 - 10%) to give compound 1-h (455 mg, 42% yield).

[0092] Synthesis of compound 1-g

[0093] A mixture of diethyl cyanomethylphosphonate (51.43 g, 290.35 mmol), triethylamine (TEA, 26.65 g, 263.95 mmol), and lithium bromide (25.21 g, 290.35 mmol) in tetrahydrofuran (THF, 700 mL) was stirred at 25 °C for 2 h. Then, cyclobutanone (18.5 g, 263.95 mmol) was added to the mixture, and some solid was separated out. Then the mixture was stirred at 25 °C for 12 h. TLC (EA (ethyl acetate):PE (petroleum ether) = 1:20, KMnO4) showed that the starting materials had been consumed and new spots were detected. The reaction was diluted with water (500 ml) and extracted with ethyl acetate (500 mL * 2). The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo, and purified by column chromatography (EA:PE = 0 - 5%, detected at 220 nm) to obtain compound 1-g (10.6 g, 43% yield).

[0094] Synthesis of compound 1-f

[0095] To a solution of compound 1-g (10.6 g, 113.82 mmol) in dichloromethane (DCM, 200 mL) at -78 °C under N₂ was slowly added 1.0 M DIBAL-H in hexane (113.8 mL, 113.8 mmol). After addition, the reaction was allowed to warm to 0 °C. Then the reaction was stirred at 0 °C for 2.5 h under N₂. TLC (EA:PE = 1:10, 254 nm) showed that the starting material had been consumed. The reaction mixture was slowly poured into 1 M H₂SO₄ solution (200 mL) at 0 °C and stirred for 30 min. Then the organic layer was separated, and the aqueous phase was extracted with DCM (200 mL × 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated in vacuo at 25 °C, and purified by silica gel column chromatography (DCM:PE = 0 - 20%) to give compound 1-f (3.1 g, 28% yield).

[0096] Synthesis of compound 1-e

[0097] To a solution of compound 1-f (3.1 g, 32.25 mmol) and 3,5-dibromo-2-methoxypyridine (8.61 g, 32.25 mmol) in THF (100 mL) at -78 °C under N₂ was slowly added n-BuLi in 2.5 M hexane (13.0 mL, 32.25 mmol). Then the reaction mixture was stirred under these conditions for 2.5 h. After the starting material was consumed (detected by LCMS), the reaction was quenched with saturated NH₄Cl (200 mL), and the mixture was extracted with ethyl acetate (200 mL × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by silica gel column chromatography (EA:PE = 0 - 26%) to give compound 1-e (3 g, 33% yield).

[0098] Synthesis of compound 1-d

[0099] To a mixture of compound 1-e (400 mg, 1.41 mmol) in glacial acetic acid (12 mL) was added 33% HBr / AcOH (0.8 mL, 4.94 mmol). Then the reaction was stirred at 80 °C for 3 h. After the starting material was consumed (detected by LCMS), the reaction mixture was directly concentrated to remove the solvent, and then saturated sodium carbonate (100 mL) was added to the residue. The mixture was extracted with ethyl acetate (60 mL × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by silica gel column chromatography (EA:PE = 0 - 50%) to give compound 1-d (110 mg, 31% yield).

[0100] Synthesis of compound 1-c

[0101] To a mixture of 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (151 mg, 0.59 mmol), compound 1-d (100 mg, 0.40 mmol) and potassium acetate (117 mg, 1.2 mmol) in 1,4-dioxane (3 mL) was added Pd(dppf)Cl2 (28 mg, 0.04 mmol). After addition, the reaction mixture was bubbled with N2 for 3 minutes and stirred at 85 °C for 3 hours. After consumption of the starting materials (detected by LCMS), the reaction mixture was concentrated and purified by silica gel column (EA:PE = 0 - 20%), to give compound 1-c (110 mg, 9.3% yield).

[0102] Synthesis of compound 1-b

[0103] To a mixture of compound 1-h (79 mg, 0.37 mmol), compound 1-c (110 mg, 0.37 mmol), sodium carbonate (120 mg, 1.11 mmol) in 1,4-dioxane (4 mL) and H2O (1 mL) was added Pd(dppf)Cl2 (27 mg, 0.037 mmol). After addition, the reaction mixture was bubbled with N2 for 3 minutes and then stirred at 95 °C for 3 hours. After consumption of the starting materials (detected by LCMS), the reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL * 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo, and purified by column (EA:PE = 0 - 30%) to give compound 1-b (51 mg, 39% yield).

[0104] Synthesis of compound 1-a

[0105] To a flask were added compound 1-b (51 mg, 0.15 mmol), (S,S)-[N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexanediamine]manganese(III) chloride (jacobsen cat(S,S)) (16 mg, 0.025 mmol) and a solution of Na2HPO4 in H2O (0.4 mL) and DCM (2 mL) (2.56 mg, 0.021 mmol). The reaction mixture was cooled to 0 °C and a solution of sodium hypochlorite (42 mg, 0.57 mmol) in H2O (1.03 mL) was slowly added to the mixture under this condition. Then the reaction was stirred at 25 °C for 12 hours. After consumption of the starting materials (by LCMS), the reaction was diluted with water (10 mL) and extracted with DCM (20 mL * 2), the organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo, and purified by silica gel column (EA:PE = 0 - 30%) to give compound 1-a (51 mg, 96% yield).

[0106] Synthesis of Compound 1

[0107] Under N2, sodium borohydride (10.5 mg, 0.28 mmol) was added to a solution of Compound 1-a (51 mg, 0.14 mmol) in THF (1.5 mL). The reaction mixture was stirred at 50 °C under N2 for 12 hours. After the consumption of the starting material (detected by LCMS), the reaction mixture was quenched with saturated NH4Cl (10 mL) and extracted with ethyl acetate (20 mL × 2). The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo, and purified by silica gel column chromatography (EA:PE = 0 - 40%) to obtain Compound 1 (13.3 mg, 26% yield). MS measured value: 370.0 [M+H] + ; 1 1H NMR (400 MHz, CDCl3): δ 8.95 (d, J = 2.2 Hz, 1H), 8.22–8.18 (m, 1H), 8.12–8.06 (m, 2H), 7.51–7.46 (m, 2H), 4.30–4.19 (m, 1H), 3.19 (dd, J = 17.0, 4.0 Hz, 1H), 2.98 (dd, J = 17.1, 3.8 Hz, 1H), 2.54–2.48 (m, 2H), 2.48–2.40 (m, 1H), 2.13–2.03 (m, 2H), 1.82–1.73 (m, 1H).

[0108] Example 2: Synthetic Route of Compound 2

[0109]

[0110] Synthesis of Compound 2-a

[0111] To a solution of 1-e (98 mg, 0.28 mmol), (R,R)-[N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexanediamine]manganese(III) chloride (jacobsen cat (R,R)) (31.7 mg, 0.05 mmol), Na2HPO4 (5.07 mg, 0.042 mmol) in H2O (0.4 mL) and DCM (cooled to 0 °C) was added a solution of sodium hypochlorite (83.8 mg, 1.12 mmol) in H2O (2.04 mL). The reaction mixture was stirred at 25 °C for 12 hours. After the consumption of the starting material (by LCMS), the reaction was diluted with water (20 mL) and extracted with DCM (30 mL × 2). The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo, and purified by silica gel column chromatography (EA:PE = 0 - 30%) to obtain Compound 2-a (65 mg, 63% yield).

[0112] Synthesis of Compound 2

[0113] Under N2, sodium borohydride (13 mg, 0.35 mmol) was added to a solution of compound 2-a (65 mg, 0.18 mmol) in THF (2 mL). Then the reaction mixture was stirred at 50 °C under N2 for 12 h. After the consumption of the starting material (detected by LCMS), the reaction mixture was quenched with saturated NH4Cl solution (10 mL), and the mixture was extracted with ethyl acetate (20 mL×2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by silica gel column chromatography (EA:PE = 0–40%) to give compound 2 (9.01 mg, 14% yield). MS found: 370.0 [M+H] + ; 1 H NMR (400 MHz, CDCl3): δ 8.96 (d, J = 2.3 Hz, 1H), 8.23–8.19 (m, 1H), 8.12–8.07 (m, 2H), 7.51–7.45 (m, 2H), 4.26 (dd, J = 9.6 Hz, 4.0 Hz, 1H), 3.19 (dd, J = 17.0 Hz, 4.1 Hz, 1H), 3.00–2.94 (m, 1H), 2.55–2.47 (m, 2H), 2.43 (dd, J = 12.3, 8.6 Hz, 1H), 2.13–2.03 (m, 2H), 1.80–1.72 (m, 1H).

[0114] Example 3: Synthetic Route of Compound 3

[0115]

[0116] Synthesis of Compound 3-g

[0117] A mixture of diethyl cyanomethylphosphonate (81.12 g, 7.94 mmol), TEA (46.29 g, 6.96 mmol) and lithium bromide (39.42 g, 458.33 mmol) in THF (400 mL) was stirred at 25 °C for 2 h. Then, cyclopentanone (35 g, 416.07 mmol) was added to the mixture, and some solids were separated out. Then the mixture was stirred at 25 °C for 12 h. TLC (EA:PE = 1:20, KMnO4) showed that the starting material had been consumed and new spots were detected. The reaction mixture was diluted with water (300 ml) and extracted with ethyl acetate (500 mL×2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo, and purified by silica gel column (EA:PE = 0–5%, detected at 220 nm) to give compound 3-g (33 g, 74% yield). 1HNMR(400MHz,CDCl3):δ5.23–5.14(m,1H),2.66–2.48(m,2H),2.47–2.37(m,2H),1.80–1.67(m,4H).

[0118] Synthesis of Compound 3-f

[0119] At -78 °C under N2, 1.0 mol / L DIBAL-H / hexane (616.9 mL, 616.9 mmol) was slowly added to a solution of Compound 3-g (33 g, 30 7.95 mmol) in DCM (300 mL). After the addition, the reaction mixture was allowed to warm to 0 °C and stirred at 0 °C for 2.5 h. TLC (EA:PE = 1:8, 254 nm) showed that the starting material had been consumed. The reaction mixture was slowly poured into 1 M H2SO4 solution (600 mL) at 0 °C and stirred for 30 min. Then the organic layer was separated, and the aqueous phase was extracted with DCM (300 mL * 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated in vacuo at 25 °C, and purified by silica gel column (DCM:PE = 0 - 20%) to give Compound 3-f (13 g, 38% yield). 1 H NMR(400MHz,CDCl3):δ9.77(d,J = 8.0Hz,1H),6.02–5.87(m,1H),2.73(t,J = 7.3Hz,2H),2.47(t,J = 7.2Hz,2H),1.82–1.72(m,2H),1.66(dd,J = 13.8,6.8Hz,2H).

[0120] Synthesis of Compound 3-e

[0121] At 0 °C under N2, i-PrMgBr (2.8 M in 2-MeTHF (6.3 mL, 17.64 mmol)) was slowly added to a solution of 5-bromo-3-iodo-2-methoxypyridine (5 g, 15.93 mmol) in THF (50 mL). The reaction mixture was stirred at 0 °C for 1 h. Then Compound 3-f (2.11 g, 19.15 mmol) was added to the reaction mixture. Then the reaction mixture was stirred under this condition for 1 h. After the starting material was consumed (determined by LCMS), the reaction mixture was quenched with saturated NH4Cl (200 mL), and the mixture was extracted with ethyl acetate (200 mL * 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by silica gel column (EA:PE = 0 - 25%) to give Compound 3-e (1.1 g, 23% yield). 11H NMR (400 MHz, CDCl3): δ 8.10 (d, J = 2.4 Hz, 1H), 7.75 (d, J = 2.4 Hz, 1H), 5.44 (dd, J = 5.4, 3.5 Hz, 2H), 3.97 (s, 3H), 2.53–2.39 (m, 2H), 2.31 (d, J = 8.4 Hz, 3H), 1.77–1.65 (m, 3H).

[0122] Synthesis of Compound 3-d

[0123] To a mixture of compound 3-e (1.1 g, 3.69 mmol) in glacial acetic acid (20 mL) was added 33% HBr / AcOH (2.1 mL, 12.92 mmol). The reaction was stirred at 80 °C for 3 h. After the raw materials were consumed (detected by LCMS), the reaction mixture was directly concentrated to remove the solvent. Saturated sodium carbonate (200 mL) was added to the residue, and the mixture was extracted with ethyl acetate (50 mL × 2). The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by silica gel column chromatography (EA:PE = 0 - 10%) to give compound 3-d (84 mg, 8.5% yield). 1 1H NMR (400 MHz, CDCl3) δ 8.01 (d, J = 2.4 Hz, 1H), 7.34 (d, J = 2.4 Hz, 1H), 6.25 (d, J = 9.8 Hz, 1H), 5.73 (d, J = 9.8 Hz, 1H), 2.26–2.14 (m, 2H), 2.05–1.97 (m, 2H), 1.70 (t, J = 4.7 Hz, 4H).

[0124] Synthesis of Compound 3-c

[0125] To a mixture of 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (149 mg, 0.59 mmol), compound 3-d (103 mg, 0.39 mmol) and potassium acetate (116 mg, 1.2 mmol) in 1,4-dioxane (5 mL) was added Pd(dppf)Cl2 (29 mg, 0.04 mmol). After addition, N2 was bubbled through the reaction solution for 3 min, and then the reaction was stirred at 85 °C for 3 h. After the raw materials were consumed (detected by LCMS), the reaction mixture was concentrated and purified by silica gel column chromatography (EA:PE = 0 - 15%) to give compound 3-c (150 mg, crude).

[0126] Synthesis of Compound 3-b

[0127] Under N2, Pd(dppf)Cl2 (18 mg, 0.024 mmol) was added to a mixture of compound 1-h (125 mg, 0.58 mmol), compound 3-c (150 mg, crude), and sodium carbonate (132 mg, 0.96 mmol) in 1,4-dioxane (5 mL) and water (1 mL). After addition, N2 was bubbled through the reaction for 3 minutes. The reaction was stirred at 100 °C for 3 hours. After consumption of the starting materials (detected by LCMS), the reaction solution was diluted with water (20 mL) and extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column (EA:PE = 0 - 30%) to give compound 3-b (72 mg, 51% yield).

[0128] Synthesis of compound 3-a

[0129] To a solution of compound 3-b (72 mg, 0.20 mmol), (S,S)-[N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexanediamine]manganese(III) chloride (jacobsen cat(S,S)) (19 mg, 0.03 mmol), and Na2HPO4 (4.3 mg, 0.03 mmol) in H2O (0.5 mL) and DCM (5 mL) (cooled to 0 °C) was slowly added a solution of sodium hypochlorite (60 mg, 0.8 mmol) in H2O (1 mL). Then, the reaction was stirred at 25 °C for 12 hours. After consumption of the starting materials (detected by LCMS), the reaction mixture was diluted with water (10 mL) and extracted with DCM (20 mL * 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by silica gel column (EA:PE = 0 - 30%) to give compound 3-a (42 mg, 56% yield).

[0130] Synthesis of compound 3

[0131] Under N2, sodium borohydride (13 mg, 0.33 mmol) was added to a mixture of compound 3-a (42 mg, 0.11 mmol) in THF (3 mL). Then the reaction was stirred at 50 °C under N2 for 12 hours. After consumption of the starting materials (determined by LCMS), the reaction was quenched with saturated NH4Cl (10 mL) and extracted with ethyl acetate (20 mL * 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by preparative HPLC to give compound 3 (3.01 mg, 7.1% yield). MS measured value was 384.1 [M+1] + ; 11H NMR (400 MHz, CDCl3): δ 8.90 (s, 1H), 8.14 (d, J = 51.1 Hz, 3H), 7.47 (s, 2H), 4.02 (s, 1H), 3.16 (d, J = 15.3 Hz, 1H), 2.98 (d, J = 15.4 Hz, 1H), 2.29 (s, 1H), 2.05 (d, J = 38.5 Hz, 4H), 1.77 (s, 3H).

[0132] Example 4: Synthetic route of compound 4

[0133]

[0134] Synthesis of compound 4-f

[0135] To a mixture of 4-chloro-3-fluorobenzonitrile (311 mg, 2 mmol) in EtOH (10 mL) was added 50% aqueous NH2OH solution (528 mg, 8 mmol). The mixture was stirred at 90 °C for 1 h, and then LCMS indicated the formation of the desired product. The mixture was concentrated in vacuo to give compound 4-f (350 mg, 93% yield).

[0136] Synthesis of compound 4-e

[0137] A mixture of 1-d (7.7 g, 30.54 mmol), Zn(CN)2 (14.3 g, 112.16 mmol), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (S-Phos, 2.5 g, 6.11 mmol) and Pd2(dba)3 (5.6 g, 6.11 mmol) in DMF (200 mL) was heated in a microwave reactor at 100 °C for 1 h. Saturated aqueous NH4Cl solution (50 mL) was added, and the mixture was partitioned between water (100 mL) and dichloromethane (100 mL), and the layers were separated. The organic layer was washed with brine (20 mL), dried (MgSO4), filtered and concentrated under reduced pressure. The crude product was purified by flash chromatography, eluting with 0 - 25% EtOAc / hexane to give compound 4-e (4.3 g, 71% yield).

[0138] Synthesis of compound 4-d

[0139] To a solution of compound 4-e (3 g, 15.13 mmol) in MeOH (80 mL) at 0 °C was slowly added concentrated H2SO4 (20 mL). After the addition, the reaction was stirred at 80 °C for 24 h. The solution was cooled to room temperature and concentrated under reduced pressure. Then DCM (100 mL) and NaOH (2 M) were added to adjust the pH to 7 - 8, and the layers were separated. The aqueous layer was treated with DCM (50 mL), the combined organic phases were dried (MgSO4), filtered and concentrated under reduced pressure. The crude product was purified by flash chromatography, eluting with 0 - 25% EtOAc / hexane to give compound 4-d (2.4 g, 69% yield).

[0140] Synthesis of compound 4-c

[0141] To a solution of compound 4-d (1.5 g, 6.49 mmol), (S,S)-[N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexanediamine]manganese(III) chloride (jacobsen cat(S,S)) (0.72 g, 1.13 mmol) and Na2HPO4 (0.12 g, 0.85 mmol) in H2O (18 mL) and DCM (80 mL) (cooled to 0 °C) was slowly added a solution of sodium hypochlorite (1.93 g, 25.96 mmol) in H2O (46 mL). Thereafter, the reaction mixture was stirred at room temperature (25 °C) for 12 h. After consumption of the starting material (detected by LCMS), the reaction was diluted with water (50 mL), extracted with DCM (50 mL * 2), the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by silica gel column (EA:PE = 0 - 30%) to give compound 4-c (850 mg, 53% yield).

[0142] Synthesis of compound 4-b

[0143] To a mixture of compound 4-c (850 mg, 3.44 mmol) in MeOH (20 mL) was added wet 10% Pd / C (250 mg). The mixture was purged with H2 twice. Then the mixture was stirred at room temperature (25 °C) for 12 h. The mixture was filtered and the cake was washed with MeOH (50 mL * 2). The filtrate was concentrated in vacuo to give the crude compound (830 mg), which was dissolved in MeOH (35 mL) and purified by supercritical fluid chromatography (SFC). The eluate was concentrated in vacuo and the residue was dried to give compound 4-b (430 mg, 50% yield).

[0144] Synthesis of compound 4-a

[0145] Sodium hydroxide (138 mg, 3.46 mmol) was added to a mixture of compound 4-b (430 mg, 1.73 mmol) in MeOH (20 mL) and H2O (8 mL). The mixture was stirred at 25 °C for 12 h. The solution was concentrated under reduced pressure and diluted with H2O (8 mL). The pH of the mixture was adjusted to 6 with HCl (2 M). The mixture was dried by lyophilization. 600 mg of compound 4-a was obtained. Some NaCl remained, and the product could be used in the next reaction without purification.

[0146] Synthesis of Compound 4

[0147] CDI (139 mg, 0.86 mmol) was added to a mixture of compound 4-a (100 mg, 0.43 mmol) in DMF (2 mL). The mixture was stirred at 25 °C for 1 h. Then 4-f (121 mg, 0.64 mmol) was added to the above mixture, and the mixture was stirred at 120 °C for 12 h. The mixture was purified by preparative HPLC to obtain compound 4 (21.07 mg, 13% yield). The MS measured value was 388.1 [M+1] + ; 1 1H NMR (400 MHz, CDCl3): δ 8.87 (d, J = 1.9 Hz, 1H), 8.13 (s, 1H), 7.92–7.79 (m, 2H), 7.53–7.44 (m, 1H), 4.19 (t, J = 3.9 Hz, 1H), 3.12 (dd, J = 17.0, 3.9 Hz, 1H), 2.96 - 2.89 (m, 1H), 2.45 (dd, J = 10.5, 6.6 Hz, 2H), 2.41 - 2.33 (m, 1H), 2.07 - 1.93 (m, 2H), 1.78 - 1.65 (m, 1H).

[0148] Example 5: Synthetic Route of Compound 5

[0149]

[0150] Synthesis of Compound 5-a

[0151] 50% aqueous NH2OH solution (1.13 g, 17.08 mmol) was added to a mixture of 4-methylbenzonitrile (500 mg, 4.27 mmol) in EtOH (10 mL). The mixture was stirred at 90 °C for 1 h. LCMS showed the formation of the desired product, and the mixture was concentrated to obtain compound 5-a (520 mg, 81% yield).

[0152] Synthesis of Compound 5

[0153] To a solution of compound 4-a (125 mg, 0.53 mmol) in DMF (3 mL) was added CDI (172 mg, 1.06 mmol). The mixture was stirred at 25 °C for 1 h. Then compound 5-a (120 mg, 0.8 mmol) was added to the above mixture and stirred at 120 °C for 3 h. LCMS showed the formation of the desired product. The mixture was extracted with EtOAc (80 mL * 3). The organic layer was concentrated in vacuo. The product was purified by preparative HPLC to give compound 5 (9.75 mg, 5.3% yield). MS found 350.1 [M+1] + ; 1 H NMR (400 MHz, CDCl3): δ 8.91 (d, J = 1.8 Hz, 1H), 8.19 (s, 1H), 8.01 (d, J = 8.1 Hz, 2H), 7.29 (d, J = 8.0 Hz, 2H), 4.24 (t, J = 3.8 Hz, 1H), 3.17 (dd, J = 17.0, 3.8 Hz, 1H), 2.97 (dd, J = 17.0, 3.7 Hz, 1H), 2.50 (t, J = 8.3 Hz, 2H), 2.42 (s, 3H), 2.13–2.02 (m, 2H), 1.82–1.70 (m, 2H).

[0154] Example 6: Synthetic route of compound 6

[0155]

[0156] Synthesis of compound 6-a

[0157] To a mixture of 1-methyl-1H-pyrazole-3-carbonitrile (100 mg, 0.93 mmol) in EtOH (5 mL) was added 50% aqueous NH2OH solution (246 mg, 3.72 mmol). The mixture was stirred at 90 °C for 1 h. LCMS showed the formation of the desired product. The mixture was concentrated in vacuo to give compound 6-a (130 mg, 99% yield).

[0158] Synthesis of compound 6

[0159] To a mixture of compound 4-a (146 mg, 0.62 mmol) in DMF (3 mL) was added CDI (201 mg, 1.24 mmol). The mixture was stirred at 25 °C for 1 h. Then 6-a (140 mg, 0.93 mmol) was added to the above mixture and stirred at 120 °C for 3 h. LCMS showed the formation of the desired product. The product was purified by preparative HPLC to give compound 6 (46.17 mg, 22% yield). MS found 340.1 [M+1] + ; 11H NMR (400 MHz, CDCl3): δ 8.98 (d, J = 2.0 Hz, 1H), 8.26 (d, J = 1.0 Hz, 1H), 7.46 (d, J = 2.2 Hz, 1H), 6.87 (d, J = 2.3 Hz, 1H), 4.22 (t, J = 3.9 Hz, 1H), 4.02 (s, 3H), 3.14 (dd, J = 17.0, 4.0 Hz, 1H), 2.93 (dd, J = 17.1, 3.9 Hz, 1H), 2.57–2.31 (m, 3H), 2.03 (ddd, J = 14.6, 10.4, 5.8 Hz, 3H), 1.84–1.63 (m, 1H).

[0160] Example 7: Synthetic route of Compound 7

[0161]

[0162] Synthesis of Compound 7-a

[0163] Hydrazine monohydrate (2 mL) was added to a solution of Compound 4-b (150 mg, 0.60 mmol) in EtOH (10 mL). The mixture was stirred at 90 °C for 12 h. The solvent was removed in vacuo, and the crude product was triturated with PE to give Compound 7-a (140 mg, 93% yield).

[0164] Synthesis of Compound 7

[0165] 4-Chlorobenzaldehyde (79 mg, 0.56 mmol) and ammonium cerium(IV) nitrate (307 mg, 0.56 mmol) were added to a solution of Compound 7-a (140 mg, 0.56 mmol) in DCM (20 mL). The mixture was stirred at 40 °C for 12 h. The mixture was purified by preparative HPLC to give Compound 7 (6.15 mg, 3% yield) as a white solid. MS calcd for 370.0 [M+1] + ; 1 1H NMR (400 MHz, CDCl3): δ 8.75 (d, J = 2.2 Hz, 1H), 8.15–8.07 (m, 1H), 8.05–7.96 (m, 2H), 7.48–7.41 (m, 2H), 4.18 (t, J = 3.9 Hz, 1H), 3.11 (dd, J = 17.1, 4.0 Hz, 1H), 2.90 (dd, J = 17.2, 3.9 Hz, 1H), 2.44 (t, J = 8.5 Hz, 2H), 2.38–2.32 (m, 1H), 2.08–1.99 (m, 2H), 1.77 (d, J = 9.1 Hz, 1H).

[0166] Example 8: Synthetic route of Compound 8

[0167]

[0168] Synthesis of Compound 8-c

[0169] To a mixture of compound 4-e (120 mg, 0.61 mmol) in EtOH (5 mL) was added 50% aqueous NH2OH solution (161 mg, 2.44 mmol). The mixture was stirred at 90 °C for 1 hour. LCMS showed the formation of the desired product. The mixture was concentrated in vacuo to give compound 8-c (130 mg, 93% yield).

[0170] Synthesis of Compound 8-b

[0171] To a mixture of 4-chlorobenzoic acid (58 mg, 0.37 mmol) in DMF (3 mL) was added CDI (120 mg, 0.74 mmol). The mixture was stirred at 25 °C for 1 hour. Then compound 8-c (130 mg, 0.56 mmol) was added and the mixture was stirred at 120 °C for 3 hours. LCMS showed the formation of the desired product. After consumption of the starting materials (determined by LCMS), the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (20 mL × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo, and purified by silica gel column (EA:PE = 0 - 20%) to give compound 8-b (30 mg, 23% yield).

[0172] Synthesis of Compound 8-a

[0173] To a mixture of compound 8-b (30 mg, 0.085 mmol), (S,S)-[N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexanediamine]manganese(III) chloride (jacobsen cat) (S,S) (8.3 mg, 0.013 mmol), and Na2HPO4 (1.7 mg, 0.012 mmol) in a mixture of H2O (0.2 mL) and DCM (2 mL) (cooled to 0 °C) was slowly added a solution of sodium hypochlorite (25 mg, 0.34 mmol) in H2O (0.5 mL). Thereafter, the reaction was stirred at 25 °C for 12 hours. After consumption of the starting materials (by LCMS), the reaction mixture was diluted with water (10 mL) and extracted with DCM (20 mL × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by silica gel column (EA:PE = 0 - 30%) to give compound 8-a (27 mg, 86% yield).

[0174] Synthesis of Compound 8

[0175] Under N2, sodium borohydride (5.76 mg, 0.152 mmol) was added to a mixture of compound 8-a (28 mg, 0.076 mmol) in THF (2 mL). The reaction was stirred at 50 °C for 12 h under N2. After consumption of the starting material (detected by LCMS), the reaction was quenched with saturated NH4Cl (10 mL) and extracted with ethyl acetate (20 mL × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by silica gel column chromatography (EA:PE = 0–40%) to give compound 8 (2.57 mg, 9.1% yield). MS calcd for: 370.0 [M+H] + ; 1 H NMR (400 MHz, CDCl3): δ 8.90 (d, J = 2.1 Hz, 1H), 8.20–8.11 (m, 3H), 7.54 (d, J = 8.6 Hz, 2H), 4.23 (t, J = 3.8 Hz, 1H), 3.18 (dd, J = 17.0, 4.1 Hz, 1H), 2.96 (dd, J = 17.1, 3.8 Hz, 1H), 2.49 (dd, J = 15.6, 6.9 Hz, 2H), 2.44–2.37 (m, 1H), 2.13–2.03 (m, 2H), 1.80–1.72 (m, 1H).

[0176] Example 9: Synthetic route of compound 9

[0177]

[0178] Synthesis of compound 9-f-1

[0179] A mixture of 5-bromo-2-chloronicotinaldehyde (1.00 g, 4.54 mmol), (triphenylphosphoranylidene)acetic acid methyl ester (1.82 g, 5.44 mmol) and THF (20 mL) was stirred at 80 °C for 2 h under N2. The reaction mixture was cooled to room temperature and concentrated. The resulting residue was purified by flash chromatography (PE / EA = 2:1) to give compound 9-f-1 (190 mg, 15% yield) as a white solid and compound 9-f-2 (877 mg, 70% yield) as a white solid. Compound 9-f-1: LC-MS (ESI): m / z = 275.9 [M+1] + ; Compound 9-f-2: MS calcd for: 275.9 [M+1] + .

[0180] Synthesis of compound 9-e-1

[0181] Under N2, an ethereal solution of CD3MgI was slowly added to a solution of compound 9-f-1 (190 mg, 0.69 mmol) in THF (10 mL) cooled to 0 °C( 2.06 mL, 2.06 mmol, 1 M). After addition, the mixture was stirred at 0 °C for 2 h. The reaction was quenched with saturated NH4Cl (30 mL), and the mixture was extracted with ethyl acetate (50 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo, and purified by flash chromatography (PE / EA = 2:1) to afford compound 9-e-1 (120 mg, 62% yield) as a white solid. MS found: 281.9 [M+H] + .

[0182] Synthesis of Compound 9-d

[0183] To a solution of compound 9-e-1 (10 mg, 0.035 mmol) in N,N-dimethylacetamide (2 mL) at room temperature was added cesium carbonate (23 mg, 0.071 mmol). The mixture was stirred at 80 °C overnight. The reaction mixture was cooled to room temperature, and H2O (10 mL) was added. The mixture was extracted with ethyl acetate (25 mL × 2), and the combined organic phases were washed with saturated NH4Cl (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo, and purified by flash chromatography (PE / EA = 2:1) to afford compound 9-d (6 mg, 69%) as a white solid. MS calcd: 246.0 [M+H] + .

[0184] Synthesis of Compound 9

[0185] According to Example 1, compound 9-d was used instead of compound 1-d to give compound 9. MS calcd: 364.1 [M+H] + .

[0186] Other examples not specifically described above were carried out in a similar manner.

[0187] For the data in Table 1, Promega's Bight-Glo TMThe luciferase assay system is used together with the TGF / SMAD signaling pathway SBE reporter gene-HEK293 cell line from BPSbioscience. On day 1, seed the SBE reporter gene-HEK293 cells at a density of 25,000 cells per well into 100 μL of growth medium without geneticin in a white clear-bottom 96-well microplate. Incubate the plate in a CO2 incubator at 37 °C for 24 hours. Refresh the wells with 60 μL of assay medium and treat the test compounds by adding 5 μL of the compound to the medium. Adjust the concentration of the test compound in the medium so that 5 μL provides the desired solution molarity. After 4 hours, add TGFβ to 10 ng / mL. Incubate the plate in a CO2 incubator at 37 °C overnight (18 hours). Replace all the medium one hour before the assay. Use the ONE-Step TM Luciferase Assay System to perform a luciferase assay by adding 100 μL of ONE-Step TM Luciferase reagent, shake for approximately 15 to 30 minutes at room temperature, and measure the luminescence using a luminometer. Normalize the signal to percentage inhibition, with 100% inhibition for non-TGF-treated cells, and process the data using Graphpad Prism.

[0188] The detection results of the luciferase screening are shown in Table 1. The symbol "***" indicates an IC 50 <100 nM, "**" indicates an IC 50 ≥100 nM but < 10 μM, and "*" indicates an IC 50 ≥10 μM.

[0189] Table 1

[0190]

[0191]

[0192] Although typical embodiments are listed for illustrative purposes, the above description and examples should not be considered as limiting the scope of the present invention. Accordingly, various modifications, adaptations, and alternatives can be made by those skilled in the art without departing from the spirit and scope of the present invention.

Claims

1. An oxygen-containing heterocyclic compound represented by Formula I, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt thereof, or a crystal form thereof: Wherein: Ar is C 6-20 aryl, C substituted with one or more R 1-1 aryl, "5- to 12-membered heteroaryl containing 1 to 4 heteroatoms selected from O, S, and N", or "5- to 12-membered heteroaryl containing 1 to 4 heteroatoms selected from O, S, and N" substituted with one or more R 6-20 aryl, "5- to 12-membered heteroaryl containing 1 to 4 heteroatoms selected from O, S, and N", or substituted with one or more R 1-2 substituted "5- to 12-membered heteroaryl containing 1 to 4 heteroatoms selected from O, S, and N"; R 1-1 and R 1-2 are independently selected from halogen, hydroxy, and C 1-6 alkyl; is a 5- or 6-membered heteroaryl containing 1 to 3 heteroatoms selected from O, S, and N; R 1 and R 2 are independently selected from hydrogen, deuterium, hydroxyl, C 1-6 alkyl, amino, and -OC 1-6 alkyl; R 3 and R 4 is a C 1-6 alkyl group substituted with deuterium, or, R 3 and R 4 together with the atom to which it is attached form a C 3-8 cycloalkyl group, a C 1-3 cycloalkyl group substituted with R 3-8 a 4- to 10-membered heterocycloalkyl group containing 1-3 heteroatoms selected from O, S, and N, a 4- to 10-membered heterocycloalkyl group containing 1-3 heteroatoms selected from O, S, and N and substituted with R 1-4 ; R 1-3 and R 1-4 are independently selected from halogen, deuterium, hydroxyl group, and C 1-6 alkyl group.

2. The oxygen-containing heterocyclic compound represented by Formula I, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt thereof, or a crystal form thereof according to claim 1, and the oxygen-containing heterocyclic compound represented by Formula I has the structure of Formula II: wherein denotes or and a mixture of; n is 1, 2, 3, 4 or 5; and Ar and as defined in Formula I.

3. The oxygen-containing heterocyclic compound represented by Formula II, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt thereof, or a crystal form thereof according to claim 2, wherein Ar is a C 1-1 aryl substituted by one or more R 6-20 groups, or a "5-12 membered heteroaryl containing 1-4 heteroatoms independently selected from O, S, and N" substituted by one or more R 1-2 groups; R 1-1 and R 1-2 are independently selected from halogen and C 1-6 alkyl; a 5- or 6-membered heteroaryl group containing 1 to 3 heteroatoms independently selected from O, S, and N; represent or and n is 2 or 3.

4. The oxygen-containing heterocyclic compound represented by Formula I, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt thereof, or a crystal form thereof according to claim 1, and the oxygen-containing heterocyclic compound represented by Formula I has the structure of Formula III: Wherein: Ar and as defined in formula I; represent or and a mixture of; and R 3 and R 4 is a C 1-6 alkyl group substituted with deuterium.

5. The oxygen-containing heterocyclic compound represented by Formula III, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt thereof, or a crystal form thereof according to claim 4, and Formula III has the following structure: wherein as defined in Formula III.

6. The oxygen-containing heterocyclic compound represented by Formula II, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt thereof, or a crystal form thereof according to claim 2, wherein Ar is a C 1-1 aryl substituted by one or more R 6-20 groups, or a "5-12 membered heteroaryl containing 1-4 heteroatoms independently selected from O, S, and N" substituted by one or more R 1-2 groups; R 1-1 and R 1-2 are independently selected from halogen and C 1-6 alkyl; a 5- or 6-membered heteroaryl group containing 1 to 3 heteroatoms independently selected from O, S, and N; indicate and n is 2 or 3.

7. The oxygen-containing heterocyclic compound represented by Formula I, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt thereof, or a crystal form thereof according to claim 1, wherein When Ar is C 6-20 Aryl or one or more R 1-1 Substituted C 6-20 When the C 6-20 The aryl group and the 1-1 Substituted C 6-20 C in aromatic groups 6-20 Aryl is C 6-10 Aryl; said C 6-10 Aryl is phenyl or naphthyl; and / or, when Ar is "a 5- to 12-membered heteroaryl containing 1 to 4 heteroatoms independently selected from O, S, and N" or "a 5- to 12-membered heteroaryl containing 1 to 4 heteroatoms independently selected from O, S, and N" substituted by one or more R 1-2 substituents, the "5- to 12-membered heteroaryl containing 1 to 4 heteroatoms independently selected from O, S, and N" and the "5- to 12-membered heteroaryl containing 1 to 4 heteroatoms independently selected from O, S, and N" substituted by one or more R 1-2 substituents, the "5- to 12-membered heteroaryl containing 1 to 4 heteroatoms independently selected from O, S, and N" is a "5-membered heteroaryl containing 2 heteroatoms selected from N"; and / or, when is a 5- or 6-membered heteroaryl containing 1-3 heteroatoms independently selected from O and N, the is a 5-membered heteroaryl containing 3 heteroatoms independently selected from O and N; and / or, when R 1-1 and R 1-2 are each independently selected from halogen, then the halogen is -F, -Cl, -Br or -I; and / or, when R 1-1 and R 1-2 are independently selected from C 1-6 alkyl, then the C 1-6 alkyl is C 1-4 alkyl, or can be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, or can be methyl; and / or, when R 3 and R 4 together with the atom to which it is attached form a C 3-8 cycloalkyl group, the C 3-8 cycloalkyl group is a C3 cycloalkyl group, a C4 cycloalkyl group, a C5 cycloalkyl group, a C6 cycloalkyl group, a C7 cycloalkyl group or a C8 cycloalkyl group; and / or, when R 3 and R 4 is a deuterium-substituted C 1-6 alkyl group, said C 1-6 alkyl group is a C 1-4 alkyl group, or may be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, or may be methyl; and / or, when R 3 and R 4 is a deuterium-substituted C 1-6 alkyl group, the deuterium-substituted C 1-6 alkyl group is -CD3.

8. The oxygen-containing heterocyclic compound represented by Formula I, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt thereof, or a crystal form thereof according to claim 1, wherein Ar is and / or, is and / or is 9. The oxygen-containing heterocyclic compound represented by Formula I, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt thereof, or a crystal form thereof according to claim 1, wherein For 10. The compound according to claim 1, its pharmaceutically acceptable salt, its deuterated derivative, its solvate, the solvate of the pharmaceutically acceptable salt, or its crystalline form, wherein R 1 is hydrogen; R 2 is selected from hydrogen, deuterium, and hydroxyl; R 3 and R 4 are C 1-6 alkyl substituted with deuterium, or, R 3 and R 4 together with the atoms to which they are attached form C 3-6 cycloalkyl; is oxadiazolyl; and Ar is selected from phenyl substituted with one or more R 1-1 and pyrazolyl substituted with one or more R 1-2 .

11. The oxygen-containing heterocyclic compound represented by Formula I according to Claim 1, its pharmaceutically acceptable salt, its deuterated derivative, its solvate, the solvate of the pharmaceutically acceptable salt, or its crystal form, wherein, The oxygen-containing heterocyclic compound represented by Formula I has any one of the following structures:

12. A pharmaceutical composition comprising Substance A and a pharmaceutically acceptable excipient, wherein Substance A is a therapeutically effective amount of the oxygen-containing heterocyclic compound represented by Formula I according to claim 1, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt thereof, or a crystal form thereof.

13. A method for inhibiting the interaction between homeodomain interacting protein kinase 2 and Smad3, the method comprising combining HIPK2 with Substance A, wherein Substance A is a therapeutically effective amount of the oxygen-containing heterocyclic compound of Formula I according to claim 1, a pharmaceutically acceptable salt thereof, a deuterated derivative thereof, a solvate thereof, a solvate of the pharmaceutically acceptable salt thereof, or a crystal form thereof.

14. A method for inhibiting Smad3 activation, wherein the method comprises contacting Smad3 with Substance A, and Substance A is a therapeutically effective amount of the oxoheterocyclic compound of Formula I according to Claim 1, its pharmaceutically acceptable salt, its deuterated derivative, its solvate, the solvate of the pharmaceutically acceptable salt, or its crystal form.

15. A method for treating a fibrotic disease, comprising administering Substance A to a subject suffering from a fibrotic disease, wherein Substance A is a therapeutically effective amount of the oxoheterocyclic compound of Formula I according to Claim 1, its pharmaceutically acceptable salt, its deuterated derivative, its solvate, the solvate of the pharmaceutically acceptable salt, or its crystal form, and the disease is renal fibrosis, cardiac fibrosis, liver fibrosis or pulmonary fibrosis.

Images