Thieno[2,3-c]pyridazin-4(1H)-one derivative and application thereof

Abstract

Disclosed are a thieno[2,3-c]pyridazin-4(1H)-one derivative as an ACC1 or ACC2 inhibitor and an application thereof in preparing a drug as an ACC1 or ACC2 inhibitor. In particular, disclosed is a compound represented by formula (II) or an isomer or pharmaceutically acceptable salt thereof. ##STR00001##

Claims

1. A compound represented by formula (II), a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof, ##STR00075## wherein, D.sub.1 is selected from —O— and —N(R.sub.6)—; R.sub.1 is selected from H, F, Cl, Br, I, OH, NH.sub.2 and C.sub.1-3 alkyl, wherein the C.sub.1-3 alkyl is optionally substituted by 1, 2 or 3 R.sub.a; R.sub.2 is selected from H, F, Cl, Br, I, OH, NH.sub.2 and C.sub.1-6 alkyl, wherein the C.sub.1-6 alkyl is optionally substituted by 1, 2 or 3 R.sub.b; R.sub.3 is selected from H, F, Cl, Br, I and C.sub.1-6 alkyl, wherein the C.sub.1-6 alkyl is optionally substituted by 1, 2 or 3 R.sub.c; or, R.sub.2 and R.sub.3 are attached together to form a ring, the ring is selected from C.sub.3-7 cycloalkyl and 4-7 membered heterocycloalkyl, the C.sub.3-7 cycloalkyl and 4-7 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 R.sub.d; R.sub.4 is selected from OH, NH.sub.2, C.sub.1-3 alkyl and C.sub.1-3 alkylamino, wherein the C.sub.1-3 alkyl and C.sub.1-3 alkylamino are optionally substituted by 1, 2 or 3 R.sub.e; each of R.sub.51, R.sub.52, R.sub.53, R.sub.54 and R.sub.55 is independently selected from H, F, Cl, Br, I, OH, NH.sub.2, C.sub.1-6 alkyl, C.sub.1-6 alkylamino and C.sub.1-6 alkoxy, wherein the C.sub.1-6 alkyl, C.sub.1-6 alkylamino and C.sub.1-6 alkoxy are optionally substituted by 1, 2 or 3 R.sub.f; R.sub.6 is selected from H, C.sub.1-6 alkyl, C.sub.1-6 alkyl-C(═O)—, C.sub.1-6 alkyl-S(═O)—, C.sub.1-6 alkyl-S(═O).sub.2— and C.sub.1-6 alkyl-O—C(═O)—, wherein the C.sub.1-6 alkyl, C.sub.1-6 alkyl-C(═O)—, C.sub.1-6 alkyl-S(═O)—, C.sub.1-6 alkyl-S(═O).sub.2— and C.sub.1-6 alkyl-O—C(═O)— are optionally substituted by R.sub.g; each of R.sub.a, R.sub.b, R.sub.c, R.sub.d, R.sub.e, R.sub.f and R.sub.g are independently selected from F, Cl, Br, I, OH, NH.sub.2 and C.sub.1-3 alkyl, wherein the C.sub.1-3 alkyl is optionally substituted by 1, 2 or 3 R; each R is independently selected from F, Cl, Br, I, OH and NH.sub.2; the 4-7 membered heterocycloalkyl contains 1, 2, 3 or 4 heteroatoms or heteroatomic groups independently selected from —NH—, —O—, —S— and N; the carbon atom labeled with “*” is a chiral carbon atom, and exists in the form of (R) or (S) single enantiomer or enriched in one enantiomer.

2. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1, wherein, each of R.sub.a, R.sub.b, R.sub.c, R.sub.d, R.sub.e, R.sub.f and R.sub.g is independently selected from F, Cl, Br, I, OH, and NH.sub.2.

3. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1, wherein, R.sub.1 is selected from H, F, Cl, Br, I, OH, NH.sub.2 and CH.sub.3.

4. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1, wherein, R.sub.2 is selected from H, F, Cl, Br, I, OH, NH.sub.2, CH.sub.3 and Et.

5. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1, wherein, R.sub.3 is selected from H, F, Cl, Br, I, CH.sub.3 and Et.

6. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1, wherein, R.sub.2 and R.sub.3 are attached together to form a ring, the ring is selected from C.sub.3-6 cycloalkyl and 5-6 membered heterocycloalkyl, the C.sub.3-6 cycloalkyl and 5-6 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 R.sub.d.

7. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 6, wherein, R.sub.2 and R.sub.3 are attached together to form a ring, the ring is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydrofuranyl, tetrahydropyranyl and piperidinyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydrofuranyl, tetrahydropyranyl and piperidinyl are optionally substituted by 1, 2 or 3 R.sub.d.

8. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 7, wherein, R.sub.2 and R.sub.3 are attached together to form a ring, the ring is selected from ##STR00076##

9. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1, wherein, R.sub.4 is selected from OH and NH.sub.2.

10. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1, wherein, each of R.sub.51, R.sub.52, R.sub.53, R.sub.54 and R.sub.55 is independently selected from H, F, Cl, Br, I, OH, NH.sub.2, C.sub.1-3 alkyl, C.sub.1-3 alkylamino and C.sub.1-3 alkoxy, wherein the C.sub.1-3 alkyl, C.sub.1-3 alkylamino and C.sub.1-3 alkoxy are optionally substituted by 1, 2 or 3 R.sub.f.

11. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 10, wherein, each of R.sub.51, R.sub.52, R.sub.53, R.sub.54 and R.sub.55 is independently selected from H, F, Cl, Br, I, OH, NH.sub.2, CH.sub.3, Et and ##STR00077##

12. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1, wherein, R.sub.6 is selected from H, C.sub.1-3 alkyl, C.sub.1-3 alkyl-C(═O)—, C.sub.1-3 alkyl-S(═O)—, C.sub.1-3 alkyl-S(═O).sub.2— and C.sub.1-4 alkyl-O—C(═O)—, wherein the C.sub.1-3 alkyl, C.sub.1-3 alkyl-C(═O)—, C.sub.1-3 alkyl-S(═O)—, C.sub.1-3 alkyl-S(═O).sub.2— and C.sub.1-4 alkyl-O—C(═O)— is optionally substituted by R.sub.g.

13. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 12, wherein, R.sub.6 is selected from H, CH.sub.3, CH.sub.3—C(═O)—, CH.sub.3—S(═O).sub.2—, CH.sub.3—O—C(═O)— and ##STR00078##

14. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1, wherein the compound is selected from ##STR00079## wherein, R.sub.1, R.sub.4, R.sub.51, R.sub.52, R.sub.53, R.sub.54 and R.sub.55 and R.sub.6 are as defined in claim 1; the carbon atom labeled with “*” is a chiral carbon atom, and exists in the form of (R) or (S) single enantiomer or enriched in one enantiomer.

15. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1, wherein the compound is selected from ##STR00080## wherein, m is 0, 1, 2 or 3; E.sub.1 is —O— or —NH—; R.sub.1, R.sub.4, R.sub.51, R.sub.52, R.sub.53, R.sub.54 and R.sub.55 are as defined in claim 1; the carbon atom labeled with “*” is a chiral carbon atom, and exists in the form of (R) or (S) single enantiomer or enriched in one enantiomer.

16. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 15, wherein the compound is selected from ##STR00081## wherein, R.sub.1, R.sub.4, R.sub.51, R.sub.52, R.sub.53, R.sub.54 and R.sub.55 are as defined in claim 15; the carbon atom labeled with “*” is a chiral carbon atom, and exists in the form of (R) or (S) single enantiomer or enriched in one enantiomer.

17. A compound represented by the following formula, a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof, ##STR00082## ##STR00083## ##STR00084## ##STR00085##

18. The compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 17, ##STR00086## ##STR00087## ##STR00088## ##STR00089##

19. A method for inhibiting ACC1 and/or ACC2 in a subject in need thereof, comprising administering the compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1 to the subject.

20. A method for treating non-alcoholic steatohepatitis and liver fibrosis in a subject in need thereof, comprising administering the compound, the stereoisomer or tautomer thereof, or the pharmaceutically acceptable salt thereof as defined in claim 1 to the subject.

Description

DETAILED DESCRIPTION OF THE EMBODIMENT

(1) The following embodiments further illustrate the present disclosure, but they are not setting any limit to the present disclosure in any sense. The present disclosure has been described in detail in the text, and its specific embodiments have also been disclosed, for one skilled person in the art, it is obvious to modify and improve the embodiments of the present disclosure without departing the spirit and scope of the present disclosure.

Reference Example 1: Fragment BB-1

(2) ##STR00022##

Synthetic Route

(3) ##STR00023##

(4) Step 1: Synthesis of Compound BB-1-2

(5) Compound BB-1-1 (25 g, 254.67 mmol) was dissolved in DCM (60 mL), a solution of sulfonyl chloride (43 mL, 430.10 mmol) in DCM (10 mL) was added dropwise at 0° C., and the mixture was stirred overnight at room temperature. After the completion of the reaction, the solvent was removed under reduced pressure to give the crude BB-1-2, which was directly used in the next step. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.62 (s, 1H), 2.14 (s, 3H).

(6) Step 2: Synthesis of Compound BB-1-3

(7) Compound BB-1-2 (40.1 g, 240.04 mmol) was dissolved in chloroform (300 mL), acetyl chloride (34.3 mL, 480.65 mmol) and aluminum trichloride (38.4 g, 287.98 mmol) were added thereto at 0° C., and the mixture was stirred overnight at room temperature. After the completion of the reaction, the reaction mixture was poured into ice water (1000 mL), stirred at room temperature for 30 minutes, and extracted with EtOAc (500 mL×2). The combined organic phase was washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, and filtered. The solvent was removed under reduced pressure, and the residue was purified by column chromatography to give BB-1-3. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 2.64 (s, 3H), 2.26 (s, 3H).

(8) Step 3: Synthesis of Compound BB-1-4

(9) Under nitrogen atmosphere, sodium hydride (12.7 g, 60% dispersed in mineral oil, 317.53 mmol) was added to toluene (200 mL), dimethyl carbonate (17.9 mL, 212.63 mmol) was added thereto, and the temperature was raised to 120° C. A solution of compound BB-1-3 (22.2 g, 106.17 mmol) in toluene (50 mL) was added dropwise over half an hour, and the reaction was allowed to continue for another half an hour. After the completion of the reaction, the mixture was quenched with water (300 mL) and the aqueous phase was extracted with EtOAc (150 mL×2). The combined organic phase was washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was purified by column chromatography to give BB-1-4. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 12.13 (s, 1H), 5.28 (s, 1H), 3.78 (s, 3H), 2.10 (s, 3H).

(10) Step 4: Synthesis of Compound BB-1-5

(11) Compound BB-1-4 (4.7 g, 17.59 mmol) and triethylamine (2.9 mL, 20.84 mmol) were added to acetonitrile (50 mL), p-toluenesulfonyl azide (4.2 g, 21.30 mmol) was added thereto at 0° C., and the reaction was allowed to continue at this temperature for 30 minutes, and then at room temperature for 2 hours. After the completion of the reaction, the mixture was quenched with water (50 mL) at 0° C., and the aqueous phase was extracted with EtOAc (25 mL×2). The combined organic phase was washed with saturated brine (10 mL×2), dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was purified by column chromatography to give BB-1-5. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.83 (s, 3H), 2.12 (s, 3H).

(12) Step 5: Synthesis of Compound BB-1-6

(13) Compound BB-1-5 (22.6 g, 77.10 mmol) was added to isopropyl ether (300 mL), a solution of tributylphosphine (20.9 mL, 84.71 mmol) in n-hexane (30 mL) was added dropwise at 0° C., and the mixture was stirred at room temperature for 2 hours. After the completion of the reaction, the solvent was removed under reduced pressure, and the residue was purified by column chromatography to give BB-1-6. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.92 (s, 3H), 2.12 (s, 3H).

(14) Step 6: Synthesis of Compound BB-1-7

(15) Compound BB-1-6 (20.1 g, 68.10 mmol) was dissolved in DCM (300 mL), Boc.sub.2O (17.8 g, 81.56 mmol) and DMAP (1.7 g, 13.92 mmol) were added thereto, and the mixture was stirred at room temperature for 12 hours. After the completion of the reaction, the solvent was removed under reduced pressure, and the residue was purified by column chromatography to give BB-1-7. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.93 (s, 3H), 2.22 (s, 3H), 1.54 (s, 9H).

(16) Step 7: Synthesis of Compound BB-1

(17) Compound BB-1-7 (26.1 g, 66.03 mmol) was dissolved in DMF (100 mL), K.sub.2CO.sub.3 (10.95 g, 79.24 mmol) was added thereto, and the reaction was allowed to run at 80° C. for 12 hours. After the completion of the reaction, water (300 mL) and HCl (1 M, 100 mL) were added, and the resulting aqueous phase was extracted with EtOAc (300 mL×2). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure, the residue was purified by column chromatography to give the target compound. LCMS: [M+H].sup.+ 258.8.

Reference Example 2: Fragment BB-2

(18) ##STR00024##

Synthetic Route

(19) ##STR00025##

(20) Step 1: Synthesis of Compound BB-2-2

(21) Compound BB-2-1 (4 g, 21.49 mmol) was dissolved in acetonitrile (50 mL), a solution of DBU (4.91 g, 32.23 mmol) and 4-acetamidobenzenesulfonyl azide (6.19 g, 25.78 mmol) in acetonitrile (10 mL) was added, and the resulting mixture was stirred overnight at room temperature. After the completion of the reaction, the solvent was removed by rotary evaporation under reduced pressure, and the residue was separated by column chromatography to give the target compound BB-2-2. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.56-7.53 (m, 1H), 7.23-7.20 (m, 1H), 7.12-7.08 (m, 1H), 3.77 (s, 3H).

(22) Step 2: Synthesis of Compound BB-2-3

(23) Compound BB-2-2 (0.99 g, 4.67 mmol) and 4-tetrahydropyranol (930 μL, 9.29 mmol) were dissolved in DCM (50 mL), rhodium acetate dimer (41 mg, 93 μmol) was added thereto, and the reaction was allowed to run at room temperature for 5 minutes. After the completion of the reaction, the solvent was removed under reduced pressure, and the residue was separated by column chromatography to give BB-2-3. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.49-7.47 (m, 1H), 7.10-7.03 (m, 1H), 6.99-6.91 (m, 1H), 5.39 (s, 1H), 3.94-3.81 (m, 2H), 3.66 (s, 3H), 3.59-3.51 (m, 1H), 3.41-3.28 (m, 2H), 1.96-1.87 (m, 1H), 1.83-1.73 (m, 1H), 1.70-1.55 (m, 2H).

(24) Step 3: Synthesis of Compound BB-2

(25) Compound BB-2-3 (5.2 g, 18.16 mmol) was dissolved in methanol (100 mL), NaBH.sub.4 (687 mg, 18.16 mmol) was added at 0° C., and the reaction was allowed to run at room temperature for 2 hours. After the completion of the reaction, water (50 mL) was added dropwise to quench the reaction at 0° C. The mixture was filtered, and the solvent was removed under reduced pressure. Water (50 mL) was added to the residue and then extracted with DCM (50 mL×2). The combined organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and dried by rotary evaporation, and the resulting residue was separated by column chromatography to give the target compound BB-2. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.52-7.49 (m, 1H), 7.14-7.12 (m, 1H), 7.05-7.01 (m, 1H), 5.05-5.01 (m, 1H), 3.99-3.96 (m, 1H), 3.92-3.90 (m, 1H), 3.72-3.70 (m, 1H), 3.57-3.32 (m, 4H), 2.26-2.23 (m, 1H), 2.05-1.96 (m, 1H), 1.79-1.63 (m, 2H), 1.60-1.54 (m, 1H).

Reference Example 3: Fragment BB-3

(26) ##STR00026##

(27) Synthetic route:

(28) ##STR00027##

(29) Step 1: Synthesis of Compound BB-3-2

(30) Compound BB-3-1 (50 g, 277.47 mmol) and NBS (49.39 g, 277.47 mmol) were dissolved in carbon tetrachloride (1 L), BPO (1.01 g, 4.16 mmol) was added thereto, and the reaction was allowed to run at 80° C. for 3 hours. After the completion of the reaction, the solvent was removed from the reaction mixture under reduced pressure to give the target compound BB-3-2, which was used directly in the next step. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.64-7.61 (m, 1H), 7.37-7.30 (m, 1H), 7.03-6.98 (m, 1H), 6.91-6.88 (m, 1H), 5.91 (s, 1H), 3.89 (s, 3H), 3.79 (s, 3H).

(31) Step 2: Synthesis of Compound BB-3-3

(32) Compound BB-3-2 (76.1 g, 293.71 mmol) and 4-tetrahydropyranol (58.8 mL, 587.24 mmol) were dissolved in DCM (1.2 L), silver oxide (68.1 g, 293.87 mmol) was added thereto, and the mixture was stirred at 25° C. for 16 hours. After the completion of the reaction, the mixture was filtered and the solvent was removed under reduced pressure. The resulting residue was purified by column chromatography to give the target compound BB-3-3. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.48-7.43 (m, 1H), 7.35-7.31 (m, 1H), 7.02-7.01 (m, 1H), 6.94-6.90 (m, 1H), 5.51 (s, 1H), 4.02-3.92 (m, 2H), 3.87 (s, 3H), 3.73 (s, 3H), 3.65-3.60 (m, 1H), 3.48-3.36 (m, 2H), 2.03-1.94 (m, 1H), 1.92-1.83 (m, 1H), 1.79-1.65 (m, 2H).

(33) Step 3: Synthesis of Compound BB-3

(34) Compound BB-3-3 (42.1 g, 150.19 mmol) was dissolved in methanol (300 mL), NaBH.sub.4 (28.4 g, 750.94 mmol) was added in batches at 0° C., and the reaction was allowed to run at room temperature for 2 hours. After the completion of the reaction, water (100 mL) was added dropwise at 0° C. to quench the reaction, and the solvent was removed under reduced pressure. Water (200 mL) was added to the residue and then extracted with DCM (250 mL×2). The combined organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, dried by rotary evaporation, and the resulting residue was separated by column chromatography to give the target compound BB-3. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.48-7.40 (m, 1H), 7.33-7.28 (m, 1H), 7.03-7.00 (m, 1H), 6.94-6.85 (m, 1H), 5.11-5.05 (m, 1H), 4.01-3.90 (m, 2H), 3.84 (s, 3H), 3.70-3.66 (m, 1H), 3.58-3.46 (m, 2H), 3.45-3.33 (m, 2H), 2.38-2.19 (m, 1H), 2.06-1.97 (m, 1H), 1.84-1.75 (m, 1H), 1.72-1.60 (m, 2H).

Example 1: WX001

(35) ##STR00028##

Synthetic Route

(36) ##STR00029## ##STR00030## ##STR00031##

(37) Step 1: Synthesis of Compound WX001-1

(38) Compound BB-1 (6.02 g, 23.27 mmol), compound BB-3 (5.87 g, 23.27 mmol) and triphenylphosphine (12.21 g, 46.54 mmol) were added to THF (250 mL), DIAD (9.0 mL, 46.29 mmol) was added thereto at 0° C., and the reaction was allowed to run at room temperature for 2 hours. After the completion of the reaction, the mixture was filtered, and the solvent was removed under reduced pressure. The residue was separated by chromatography column to give the target compound WX001-1. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.56-7.54 (m, 1H), 7.38-7.34 (m, 1H), 7.08-7.04 (m, 1H), 6.93-6.91 (m, 1H), 5.44-5.41 (m, 1H), 4.43-4.39 (m, 1H), 4.17-4.12 (m, 1H), 4.01 (s, 3H), 3.90 (s, 3H), 3.73-3.65 (m, 1H), 3.59-3.49 (m, 1H), 3.31-3.28 (m, 1H), 3.27-3.20 (m, 2H), 2.62 (s, 3H), 1.78-1.66 (m, 1H), 1.64-1.48 (m, 2H), 1.29-1.23 (m, 1H).

(39) Step 2: Synthesis of Compound WX001-2

(40) NaBH.sub.4 (1.73 g, 45.64 mmol) was dissolved in methanol (70 mL), compound WX001-1 (4.5 g, 9.13 mmol) was added in batches at 0° C., and the reaction was allowed to run at 50° C. for 1 hour. The reaction mixture was quenched with water (20 mL) at 0° C., and extracted with DCM (25 mL×2). The organic phase was combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was used directly in the next step.

(41) Step 3: Synthesis of Compound WX001-3

(42) Under nitrogen atmosphere, compound WX001-2 (4.2 g, 9.03 mmol) and phosphorus tribromide (940 μL, 9.90 mmol) were dissolved in DCM (50 mL), and the reaction was allowed to run at room temperature for half an hour. After the completion of the reaction, the solvent was removed under reduced pressure, and the resulting residue was separated by chromatography column to give the target compound WX001-3. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.56-7.54 (m, 1H), 7.37-7.33 (m, 1H), 7.07-7.03 (m, 1H), 6.93-6.91 (m, 1H), 5.43-5.40 (m, 1H), 4.64 (d, J=9.2 Hz, 1H), 4.57 (d, J=9.2 Hz, 1H), 4.38-4.33 (m, 1H), 4.07-4.04 (m, 1H), 3.91 (s, 3H), 3.75-3.65 (m, 1H), 3.63-3.52 (m, 1H), 3.42-3.25 (m, 3H), 2.62 (s, 3H), 1.74-1.64 (m, 1H), 1.59-1.54 (m, 2H), 1.24-1.20 (m, 1H).

(43) Step 4: Synthesis of Compound WX001-4

(44) Compound WX001-3 (3.8 g, 7.20 mmol) was dissolved in DMF (10 mL), KCN (2 g, 30.71 mmol) was added thereto, and the reaction was allowed to run at room temperature for 2 hours. TLC detected that the raw material was still remained, KCN (1.6 g, 24.57 mmol) was added, and the reaction was allowed to continue for another 2.5 hours. After the completion of the reaction, the mixture was quenched with water (50 mL) at 0° C., and extracted with EtOAc (25 mL×2). The organic phase was combined, washed with saturated brine (25 mL), and dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The resulting residue was separated by chromatography column to give the target compound WX001-4. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.50-7.37 (m, 1H), 7.31-7.22 (m, 1H), 6.98-6.94 (m, 1H), 6.86-6.84 (m, 1H), 5.39-5.36 (m, 1H), 4.41-4.24 (m, 1H), 4.05-3.99 (m, 1H), 3.98-3.75 (m, 5H), 3.74-3.63 (m, 1H), 3.56-3.46 (m, 1H), 3.33-3.29 (m, 1H), 3.28-3.15 (m, 2H), 2.51 (s, 3H), 1.71-1.61 (m, 1H), 1.59-1.41 (m, 2H), 1.16-1.04 (m, 1H).

(45) Step 5: Synthesis of Compound WX001-5

(46) Compound WX001-4 (2.83 g, 5.97 mmol), 2-(tri-n-butylstannyl)oxazole (5.35 g, 14.93 mmol) were dissolved in toluene (100 mL), and tetrakis(triphenylphosphine)palladium (2.07 g, 1.79 mmol) was added thereto. The reaction mixture was purged with nitrogen for 3 times and raised to 120° C., and the reaction was allowed to run for 1 hour. The reaction mixture was reduced to room temperature and the solvent was removed under reduced pressure. The residue was dissolved in DCM (30 mL) and quenched with saturated potassium fluoride (30 mL). The mixture was extracted with DCM (30 mL), the combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was separated by chromatography column to give the target compound WX001-5. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.71 (s, 1H), 7.48-7.45 (m, 1H), 7.27-7.26 (m, 1H), 7.21 (s, 1H), 6.99-6.95 (m, 1H), 6.86-6.84 (m, 1H), 5.45-5.41 (m, 1H), 4.42-4.38 (m, 1H), 4.14-4.09 (m, 1H), 3.94-3.82 (m, 5H), 3.71-3.62 (m, 1H), 3.54-3.43 (m, 1H), 3.30-3.35 (m, 1H), 3.27-3.12 (m, 2H), 2.95 (s, 3H), 1.70-1.60 (m, 1H), 1.57-1.47 (m, 2H), 1.12-1.09 (m, 1H).

(47) Step 6: Synthesis of Compound WX001-6

(48) Compound WX001-5 (0.1 g, 197.41 μmol) was dissolved in THF (10 mL), LiHMDS (1M, 590 μL, 590 μmol) was added dropwise at −65° C., and the reaction was allowed to run for half an hour. Then 1,4-dibromobutane (70 μL, 580.32 μmol) was added dropwise, and the reaction was allowed to continue at room temperature for half an hour. After the completion of the reaction, water (10 mL) was added dropwise to quench the reaction at 0° C., and extracted with EtOAc (10 mL×2). The organic phase was combined and washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the solvent was removed under reduced pressure. The residue was separated by preparation plate to give the target compound WX001-6. LCMS (5-95/1.5 min): 0.973 min, [M+H].sup.P=561.1.

(49) Step 7: Synthesis of Compound WX001-7

(50) Compound WX001-6 (140 mg, 249.70 μmol) was dissolved in benzyl alcohol (1 mL), a solution of hydrogen chloride in 1,4-dioxane (4 M, 62 μL, 248 μmol) was added under nitrogen atmosphere, and the reaction was allowed to run at 50° C. for 2 hours. After the completion of the reaction, the solvent was removed under reduced pressure, and the resulting residue was separated by preparative HPLC to give the target compound WX001-7 (hydrochloric acid condition). LCMS (5-95/1.5 min): 1.095 min, [M+1-1].sup.+=670.1.

(51) Step 8: Synthesis of Compound WX001

(52) Compound WX001-7 (0.14 g, 209.02 μmol) was dissolved in MeOH (10 mL), and 10% Pd/C (30 mg) was added under nitrogen atmosphere. The reaction mixture was purged with hydrogen for 3 times, and the reaction was allowed to run under hydrogen atmosphere (30 Psi) at 30° C. for 2 hours. The reaction mixture was filtered, and the solvent was removed under reduced pressure to give a residue. The residue was separated by preparative chromatography (hydrochloric acid condition) to give the target compound WX001. The compound WX001 was analyzed by supercritical fluid chromatography (column: Chiralpak AD-3 100×4.6 mm I.D., 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B from 5% to 40% in 4.5 minutes, 40% for 2.5 minutes, back to 5% equilibration for 1 minute; flow rate: 2.8 mL/min; column temperature: 40° C.; wavelength: 220 nm) as racemic compounds. Chiral isomers WX001A and WX001B were separated, and their retention time was 3.954 min and 4.388 min respectively.

(53) Referring to the synthesis method of steps 6-8 in the Embodiment 1, the example in the following table was synthesized using different halide fragments in step 6. The structure in the table also represents its possible isomers.

(54) TABLE-US-00001 TABLE 1 Compound structure of each example Example Halide fragment Structure Compound 2 — WX002 WX002A or WX002B WX002B or WX002A 3 Hydrolyzing the nitrile group in WX001-5 WX003 4 MeI WX004 WX004A or WX004B WX004B or WX004A 5 0 WX005 6 WX006 7 WX007 WX007A or WX007B WX007B or WX007A 8 WX008 WX008A or WX008B 0 WX008B or WX008A 9 WX009 WX009A or WX009B WX009B or WX009A 10 WX010 16 MeI; The methoxybenzene in BB-3 intermediate was replaced with p- fluoromethoxybenzene WX016 WX016A or WX016B WX016B or WX016A

(55) TABLE-US-00002 TABLE 2 NMR and MS data of each example Example Compound NMR MS m/z: 1 SFC detection method: Column: Chiralpak AD-3 100 × 4.6 mm I.D., 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B from 5% to 40% in 4.5 minutes, 40% for 2.5 minutes, back to 5% equilibrium for 1 minute; flow rate: 2.8 mL/min; column temperature: 40° C.; wavelength: 220 nm. WX001 .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.69 (s, 1H), 7.47-7.45 580.1 (m, 1H), 7.26-7.24 (m, 1H), 7.20 (s, 1H), 6.99-6.95 (M + H).sup.+ (m, 1H), 6.84-6.82 (m, 1H), 5.42-5.38 (m, 1H), 4.38- 4.34 (m, 1H), 4.13-4.07 (m, 1H), 3.80 (s, 3H), 3.70- 3.60 (m, 1H), 3.47-3.38 (m, 1H), 3.35-3.08 (m, 3H), 2.95 (s, 3H), 2.40-2.29 (m, 2H), 2.20-2.15 (m, 2H), 1.77-1.75 (m, 2H), 1.62-1.48 (m, 5H), 1.12-1.09(m, 1H). A pair of racemates, with a ratio of 1:1, detected by SFC. WX001A SFC retention time 3.954 min .sup.1H NMR (400 MHz, CDCl.sub.3,) δ 7.70 (s, 1H), 7.47-7.44 (m, 1H), 7.26-7.25 (m, 1H), 7.20 (s, 1H), 6.99-6.97 (m, 1H), 6.85-6.83 (m, 1H), 5.42-5.38 (m, 1H), 4.40- 4.35 (m, 1H), 4.14-4.08 (m, 1H), 3.81 (s, 3H), 3.71- 3.62 (m, 1H), 3.46-3.41 (m, 1H), 3.31-3.25 (m, 1H), 3.24-3.18 (m, 1H), 3.11-3.17 (m, 1H), 2.96 (s, 3H), 2.43-2.40 (m, 2H), 2.17-2.16 (m, 2H), 1.78-1.70 (m, 2H), 1.62-1.49 (m, 3H), 1.48-1.46 (m, 2H), 1.12- 1.08 (m, 1H). WX001B SFC retention time 4.388 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.70 (s, 1H), 7.46-7.44 (m, 1H), 7.27-7.24 (m, 1H), 7.20 (s, 1H), 6.99-6.97 (m, 1H), 6.85-6.83 (m, 1H), 5.41-5.38 (m, 1H), 4.39- 4.35 (m, 1H), 4.14-4.08 (m, 1H), 3.81 (s, 3H), 3.71- 3.62 (m, 1H), 3.46-3.41 (m, 1H), 3.31-3.25 (m, 1H), 3.24-3.18 (m, 1H), 3.11-3.17 (m, 1H), 2.96 (s, 3H), 2.42-2.40 (m, 2H), 2.18-2.16 (m, 2H), 1.77-1.75 (m, 2H), 1.62-1.58 (m, 3H), 1.48-1.46 (m, 2H), 1.11- 1.08 (m, 1H). 2 WX002 .sup.1H NMR (400 MHz, CDCl.sub.3) δ7.79 (s, 1H), 7.58-7.56 525.1 (m, 1H), 7.38-7.34 (m, 1H), 7.30 (s, 1H), 7.21 (bs, (M + H).sup.+ 1H), 7.08-7.04 (m, 1H), 6.94-6.92 (m, 1H), 5.52- 5.48 (m, 1H), 5.39 (bs, 1H), 4.50-4.45 (m, 1H), 4.18- 4.12 (m, 1H), 3.93 (s, 3H), 3.84-3.82 (m, 2H), 3.75- 3.67 (m, 1H), 3.54-3.45 (m, 1H), 3.39-3.36 (m, 1H), 3.35-3.22 (m, 2H), 3.06 (s, 3H), 1.71-1.72 (m, 1H), 1.59-1.50 (m, 2H), 1.18-1.15 (m, 1H). 3 SFC detection method: column: Chiralcel OJ-3 100 × 4.6 mm I.D., 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in methanol; gradient: B from 5% to 40% in 4.5 minutes, 40% for 2.5 minutes, back to 5% equilibrium for 1 minute; flow rate: 2.8 mL/min; column temperature: 40° C.; wavelength: 220 nm. WX003 The crude product was directly purified by SFC. As 525.9 racemates with a ratio of 1:1, detected by SFC. (M + H).sup.+ WX003A SFC retention time 2.343 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.73 (s, 1H), 7.47-7.45 (m, 1H), 7.28-7.26 (m, 1H), 7.22 (s, 1H), 6.99-6.97 (m, 1H), 6.86-6.84 (m, 1H), 5.41-5.38 (m, 1H), 4.43- 4.39 (m, 1H), 4.15-4.09 (m, 1H), 3.91 (s, 2H), 3.84 (s, 3H), 3.68-3.60 (m, 1H), 3.48-3.40 (m, 1H), 3.35- 3.26 (m, 1H), 3.25-3.12 (m, 2H), 2.98 (s, 3H), 1.61- 1.57 (m, 1H), 1.52-1.41 (m, 2H), 1.08-1.05 (m, 1H). WX003B SFC retention time 2.685 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.72 (s, 1H), 7.48-7.46 (m, 1H), 7.28-7.26 (m, 1H), 7.22 (s, 1H), 6.99-6.95 (m, 1H), 6.86-6.84 (m, 1H), 5.41-5.38 (m, 1H), 4.42- 4.38 (m, 1H), 4.14-4.09 (m, 1H), 3.89 (s, 2H), 3.84 (s, 3H), 3.68-3.60 (m, 1H), 3.48-3.40 (m, 1H), 3.35- 3.26 (m, 1H), 3.25-3.12 (m, 2H), 2.97 (s, 3H), 1.61- 1.60 (m, 1H), 1.49-1.42 (m, 2H), 1.08-1.06 (m, 1H). 4 SFC detection method: column: Chiralpak AD-3 100 × 4.6 mm I.D., 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B from 5% to 40% in 2.5 minutes, 40% for 2 minutes, back to 5% equilibrium for 1 minute. Flow rate: 2.5 mL/min; column temperature: 40° C.; wavelength: 220 nm. WX004 The crude product obtained after hydrogenation was 554.0 directly separated by SFC to give the enantiomers. (M + H).sup.+ As racemates with a ratio of 1:1, detected by SFC. WX004A SFC retention time 3.636 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.69 (s, 1H), 7.47-7.45 (m, 1H), 7.26-7.24 (m, 1H), 7.22 (s, 1H), 6.98-6.94 (m, 1H), 6.84-6.82 (m, 1H), 5.42-5.40 (m, 1H), 4.38- 4.34 (m, 1H), 4.15-4.10 (m, 1H), 3.78 (s, 3H), 3.71- 3.63 (m, 1H), 3.50-3.42 (m, 1H), 3.35-3.32 (m, 1H), 3.28-3.13 (m, 2H), 2.95 (s, 3H), 1.64-1.62 (m, 1H), 1.54-1.43 (m, 8H), 1.16-1.14 (m, 1H). WX004B SFC retention time 3.940 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.69 (s, 1H), 7.47-7.45 (m, 1H), 7.26-7.24 (m, 1H), 7.22 (s, 1H), 6.98-6.95 (m, 1H), 6.84-6.82 (m, 1H), 5.43-5.40 (m, 1H), 4.39- 4.34 (m, 1H), 4.15-4.10 (m, 1H), 3.78 (s, 3H), 3.71- 3.63 (m, 1H), 3.50-3.42 (m, 1H), 3.35-3.32 (m, 1H), 3.28-3.13 (m, 2H), 2.95 (s, 3H), 1.64-1.62 (m, 1H), 1.54-1.43 (m, 8H), 1.19-1.16 (m, 1H). 5 WX005 .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.71 (s, 1H), 7.45-7.42 552.1 (m, 1H), 7.29-7.24 (m, 1H), 7.21 (s, 1H), 6.99-6.93 (M + H).sup.+ (m, 1H), 6.86-6.80 (m, 1H), 5.36-5.34 (m, 1H), 4.39- 4.36 (m, 1H), 4.18-4.10 (m, 1H), 3.82 (s, 3H), 3.69- 3.63 (m, 1H), 3.49-3.42 (m, 1H), 3.34-3.31 (m, 1H), 3.26-3.13 (m, 2H), 2.98 (s, 3H), 1.61-1.55 (m, 2H), 1.47-1.42 (m, 2H), 1.20-1.12 (m, 4H). 6 WX006 .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.81 (s, 1H), 7.52-7.51 566.2 (m, 1H), 7.37-7.31 (m, 1H), 7.31 (s, 1H), 7.07-7.03 (m, (M + H).sup.+ 1H), 6.93-6.91 (m, 1H), 5.52-5.48 (m, 1H), 4.54-4.50 (m, 1H), 4.30-4.24 (m, 1H), 3.88 (s, 3H), 3.78-3.69 (m, 1H), 3.58-3.51 (m, 1H), 3.45-3.37 (m, 1H), 3.36- 3.21 (m, 2H), 3.05 (s, 3H), 3.00-2.89 (m, 2H), 2.68- 2.56 (m, 2H), 2.21-2.09 (m, 1H), 1.83-1.81 (m, 1H), 1.72-1.70 (m, 2H), 1.51-1.48 (m, 1H), 1.24-1.17 (m, 1H). 7 SFC detection method: Column: Chiralpak AD-3 50 × 3 mm I.D., 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in isopropanol; gradient: B from 5% to 40% within 2.5 minutes, 40% for 0.35 min, from 40% back to 5% in 0.15 minute. Flow rate: 2.5 mL/min; column temperature: 40° C.; Wavelength: 220 nm. WX007 The crude product was directly separated by SFC to 594.0 give the enantiomers. As racemates with a ratio of 1:1, (M + H).sup.+ detected by SFC. WX007A SFC retention time 1.875 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.71 (s, 1H), 7.47-7.45 (m, 1H), 7.30-7.25 (m, 1H), 7.21 (s, 1H), 6.99-6.96 (m, 1H), 6.85-6.83 (m, 1H), 5.45-5.42 (m, 1H), 4.43- 4.38 (m, 1H), 4.15-4.10 (m, 1H), 3.80 (s, 3H), 3.68- 3.64 (m, 1H), 3.47-3.42 (m, 1H), 3.34-3.30 (m, 1H), 3.27-3.20 (m, 1H), 3.19-3.12 (m, 1H), 2.96 (s, 3H), 2.17-2.05 (m, 4H), 1.69-1.63 (m, 2H), 1.53-1.45 (m, 5H), 1.32-1.41 (m, 2H), 1.12-1.09 (m, 1H). WX007B SFC retention time 2.177 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.80 (s, 1H), 7.56-7.54 (m, 1H), 7.39-7.33 (m, 1H), 7.30 (s, 1H), 7.09-7.04 (m, 1H), 6.94-6.92 (m, 1H), 5.54-5.51 (m, 1H), 4.52- 4.48 (m, 1H), 4.22-4.19 (m, 1H), 3.89 (s, 3H), 3.79- 3.72 (m, 1H), 3.56-3.51 (m, 1H), 3.45-3.37 (m, 1H), 3.35-3.29 (m, 1H), 3.27-3.21 (m, 1H), 3.05 (s, 3H), 2.26-2.07 (m, 4H), 1.74-1.73 (m, 2H), 1.65-1.54 (m, 5H), 1.51-1.47 (m, 2H), 1.23-1.17 (m, 1H). 8 SFC detection method: column: Chiralpak AD-3 100 × 4.6 mm I.D., 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in isopropanol; gradient: B from 5% to 40% in 2.5 minutes, 40% for 0.35 minute, from 40% to 5% in 0.15 minute. Flow rate: 2.5 mL/min; column temperature: 40° C.; wavelength: 220 nm. WX008 The crude product was directly separated by SFC to 596.1 give the enantiomers. As racemates with a ratio of 1:1, (M + H).sup.+ detected by SFC. WX008A SFC retention time 2.017 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.72 (s, 1H), 7.46-7.44 (m, 1H), 7.31-7.26 (m, 1H), 7.22 (s, 1H), 7.00-6.96 (m, 1H), 6.86-6.84 (m, 1H), 5.43-5.40 (m, 1H), 4.44- 4.39 (m, 1H), 4.18-4.13 (m, 1H), 3.86-3.79 (m, 5H), 3.79-3.61 (m, 4H), 3.48-3.42 (m, 1H), 3.34-3.30 (m, 1H), 3.27-3.14 (m, 2H), 2.95 (s, 3H), 2.48-2.41 (m, 2H), 2.21-2.11 (m, 2H), 1.66-1.62 (m, 1H), 1.48-1.45 (m, 2H), 1.12-1.03 (m, 1H). WX008B SFC retention time 2.336 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.72 (s, 1H), 7.47-7.44 (m, 1H), 7.32-7.26 (m, 1H), 7.22 (s, 1H), 7.00-6.96 (m, 1H), 6.86-6.84 (m, 1H), 5.43-5.38 (m, 1H), 4.45- 4.40 (m, 1H), 4.16-4.14 (m, 1H), 3.86-3.82 (m, 1H), 3.80 (s, 3H), 3.49-3.43 (m, 1H), 3.34-3.30 (m, 1H), 3.26-3.20 (m, 1H), 3.19-3.12 (m, 1H), 2.95 (s, 3H), 2.43-2.41 (m, 2H), 2.16-2.14 (m, 2H), 1.67-1.65 (m, 1H), 1.57-1.45 (m, 2H), 1.12-1.06 (m, 1H). 9 SFC detection method: column: Chiralcel AD-3 100 × 4.6 mm I.D., 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B from 5% to 40% in 4.5 minutes, 40% for 2.5 minutes, back to 5% equilibrium for 1 minute; flow rate: 2.8 mL/min; column temperature: 40° C.; wavelength: 220 nm. WX009 The crude product was directly separated by SFC to 562.1 give the enantiomers. As racemates with a ratio of 1:1, (M + H).sup.+ detected by SFC. WX009A SFC retention time is 3.273 min. .sup.1H NMR (400 MHz, CDCl.sub.3,) δ 7.63 (s, 1H), 7.45-7.49 (m, 1H), 7.25-7.21 (m, 1H), 7.12 (s, 1H), 6.95-6.91 (m, 1H), 6.81-6.79 (m, 1H), 5.41-5.39 (m, 1H), 4.40-4.37 (m, 1H), 4.14-4.13 (m, 1H), 3.79 (s, 3H), 3.64-3.59 (m, 1H), 3.42 (s, 3H), 3.30-3.28 (m, 1H), 3.19-3.12 (m, 1H), 2.82-2.78 (m, 2H), 1.57-1.49 (m, 2H), 1.41-1.39 (m, 1H), 1.19-1.17 (m, 1H). WX009B SFC retention time is 3.674 min. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.77 (s, 1H), 7.56-7.54 (m, 1H), 7.37-7.34 (m, 1H), 7.25 (s, 1H), 7.04-7.02 (m, 1H), 6.93-6.91 (m, 1H), 5.53-5.51 (m, 1H), 4.55- 4.52 (m, 1H), 4.30-4.25 (m, 1H), 3.90 (s, 3H), 3.75- 3.70 (m, 1H), 3.60-3.58 (m, 1H), 3.45-3.39 (m, 1H), 3.31-3.25 (m, 1H), 2.96 (s, 3H), 3.30-3.28 (m, 1H), 3.19-3.12 (m, 1H), 2.82-2.78 (m, 2H), 1.57-1.41 (m, 2H), 1.25-1.17 (m, 1H). 10 WX010 .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.76-8.70 (d, 1H), 595.1 8.30 (s, 1H), 7.51-7.49 (m, 1H), 7.46 (s, 1H), 7.38- (M + H).sup.+ 7.34 (m, 1H), 7.08-7.05 (m, 2H), 5.41-5.38 (m, 1H), 4.50-4.45 (m, 1H), 4.25-4.19 (m, 1H), 3.88 (s, 3H), 3.63-3.59 (m, 1H), 3.28-3.15 (m, 5H), 3.14-3.08 (m, 2H), 2.90 (s, 3H), 2.39-2.21 (m, 4H), 1.70-1.62 (m, 1H), 1.57-1.48 (m, 1H), 1.40-1.34 (m, 1H), 1.22- 1.17 (m, 1H), 1.05-0.94 (m, 1H) 16 SFC detection method: column: Lux 3u Cellulose-2 150*4.6 mm 572.1 I.D., 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% (M + H).sup.+ diethylamine in ethanol; gradient: B from 5% to 40% in 4.5 minutes, 40% for 2.5 minutes, back to 5% equilibrium for 1 minute; flow rate: 2.5 mL/min; column temperature: 40° C.; wavelength: 220 nm. WX016A The SFC retention time is 3.725 min. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.69 (s, 1H), 7.20-7.22 (m, 2H), 6.93-6.91 (m, 1H), 6.77-6.73 (m, 1H), 5.37-5.36 (m, 1H), 4.40-4.33 (m, 1H), 4.13-4.10 (m, 1H), 3.75 (s, 3H), 3.70-3.67 (m, 1H), 3.48-3.43 (m, 1H), 3.32-3.17 (m, 3H), 2.94 (s, 3H), 1.68- 1.62 (m, 1H), 1.59-1.53 (m, 1H), 1.50 (s, 3H), 1.45 (s, 3H), 1.18-1.13 (m, 1H), 0.83-0.79 (m, 1 H). WX016B The SFC retention time is 6.972 min. .sup.1H NMR (400 MHz, CDCl.sub.3) δ7.74 (s, 1H), 7.24-7.23 (m, 2H), 7.00-6.97 (m, 1H), 6.82-6.79 (m, 1H), 5.44-5.41 (m, 1H) 4.41-4.37 (m, 1H), 4.17-4.14 (m, 1H), 3.81 (s, 3 H), 3.76-3.73 (m, 1H), 3.56- 3.49 (m, 1H), 3.39-3.23 (m, 3H), 2.97 (s, 3H) 1.73- 1.70 (m, 1H), 1.66-1.59 (m, 1H), 1.54 (s, 3H), 1.49 (s, 3H), 1.25-1.16 (m, 1H) 0.83-0.85 (m, 1H).

Example 11: WX011

(56) ##STR00060##

Synthetic Route

(57) ##STR00061## ##STR00062## ##STR00063##

(58) Step 1: Synthesis of Compound WX011-1

(59) Compound BB-1 (20 g, 77.32 mmol) was dissolved in DMF (270 mL), NaH (4.02 g, 60% dispersed in mineral oil, 100.51 mmol) was added thereto, and the mixture was stirred at 0° C. for 30 minutes. Then SEM-Cl (15 mL, 84.75 mmol) was added, and the reaction was allowed to run at room temperature for 1 hour. The reaction mixture was quenched with water (800 mL), and extracted with EtOAc (400 mL×2). The organic phase was combined and washed with saturated brine (100 mL×4), dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The resulting residue was separated by chromatography column to give the target compound WX011-1.

(60) .sup.1HNMR (400 MHz, CDCl.sub.3) δ 5.52 (s, 2H), 3.99 (s, 3H), 3.66-3.62 (m, 2H) 2.59 (s, 3H), 1.00-0.96 (m, 2H), 0.00 (s, 9H).

(61) Step 2: Synthesis of Compound WX011-2

(62) Compound WX011-1 (10 g, 25.71 mmol) was dissolved in methanol (100 mL), lithium borohydride (2.8 g, 128.55 mmol) was added thereto, and the reaction was allowed to run at room temperature for 2 hours. The reaction mixture was quenched with water (200 mL) and the methanol was removed under reduced pressure. The mixture was extracted with EtOAc (300 mL×2), and the organic phase was combined, washed with saturated brine (50 mL×2), dried over anhydrous sodium sulfate, and filtered. The solvent was removed under reduced pressure, and the resulting residue was used directly in the next step. .sup.1HNMR (400 MHz, CDCl.sub.3) δ 5.48 (s, 2H), 4.77 (s, 2H), 3.64-3.60 (m, 2H), 2.58 (s, 3H), 1.00-0.95 (m, 2H), 0.01 (s, 9H).

(63) Step 3: Synthesis of Compound WX011-3

(64) Compound WX011-2 (8 g, 22.16 mmol) and triethylamine (6.2 mL, 44.33 mmol) were dissolved in DCM (100 mL), MsCl (2.3 mL, 29.68 mmol) was added thereto at 0° C., and the reaction was allowed to run for 1 hour after the addition was completed. The reaction mixture was quenched with ice water (100 mL), and extracted with DCM (60 mL×2). The organic phase was combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The resulting residue was used directly used in the next step.

(65) Step 4: Synthesis of Compound WX011-4

(66) Compound WX011-3 (8.5 g, 19.36 mmol) was dissolved in DMF (100 mL), NaCN (4.07 g, 83.04 mmol) was added thereto, and the reaction was allowed to run at room temperature for 2 hours. After the completion of the reaction, the reaction was quenched by water (200 mL) and extracted with EtOAc (200 mL×3). The organic phase was combined, washed with saturated brine (100 mL×4), dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The resulting residue was separated by chromatography column to give the target compound WX011-4. .sup.1HNMR (400 MHz, DMSO-d.sub.6) δ 5.51 (s, 2H), 3.90 (s, 2H), 3.66-3.62 (m, 2H), 2.56 (s, 3H), 0.99-0.95 (m, 2H), 0.00 (s, 9H).

(67) Step 5: Synthesis of Compound WX011-5

(68) Compound WX011-4 (2.5 g, 6.76 mmol), 2-(tri-n-butylstannyl)oxazole (6.05 g, 16.89 mmol) were dissolved in toluene (30 mL), and tetrakis(triphenylphosphine)palladium (1.56 g, 1.35 mmol) was added thereto. The reaction mixture was purged with nitrogen for 3 times and raised to 120° C., and the reaction was allowed to run for 4 hours. After cooling to room temperature, the reaction was quenched with saturated potassium fluoride (20 mL). Water (80 mL) was added and extracted with EtOAc (100 mL×2). The organic phase were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The residue was separated by chromatography column to give the target compound WX011-5. .sup.1HNMR (400 MHz, CDCl.sub.3) δ 7.77 (s, 1H), 7.29 (s, 1H), 5.60 (s, 2H), 3.94 (s, 2H), 3.71-3.67 (m, 2H), 3.02 (s, 3H), 1.02-0.98 (m, 2H), 0.01 (s, 9H).

(69) Step 6: Synthesis of Compound WX011-6

(70) Compound WX011-5 (1.9 g, 4.72 mmol) and methyl iodide (1.6 mL, 25.50 mol) were dissolved in THF (20 mL), potassium tert-butoxide solution (1 M, 14.2 mL, 14.2 mmol) was added dropwise at 0° C., and the reaction was allowed to run at room temperature for 1 hour. The reaction was quenched with water (100 mL), and extracted with EtOAc (100 mL×2). The organic phase was combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the solvent was removed under reduced pressure. The residue was used directly in the next step. .sup.1HNMR (400 MHz, CDCl.sub.3) δ 7.75 (s, 1H), 7.29 (s, 1H), 5.54 (s, 2H), 3.70-3.65 (m, 2H), 3.02 (s, 3H), 1.64 (s, 6H), 1.00-0.96 (m, 2H), 0.01 (s, 9H).

(71) Step 7: Synthesis of Compound WX011-7

(72) Compound WX011-6 (1 g, 2.32 mmol) was added to a solution of TBAF (1 M, 15 mL, 15 mmol) in THF, and the reaction was allowed to run at room temperature for 1 hour. After the completion of the reaction, the reaction was quenched with water (80 mL), and extracted with EtOAc (100 mL×2). The combined organic phase was washed with water (50 mL×5), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the solvent in the filtrate was removed under reduced pressure. The resulting residue was directly used in the next step. .sup.1HNMR (400 MHz, CDCl.sub.3) δ 7.72 (s, 1H), 7.24 (s, 1H), 2.95 (s, 3H), 1.81 (s, 6H).

(73) Step 8: Synthesis of Compound WX011-8

(74) Compound WX011-7 (0.56 g, 1.86 mmol), compound WX011-7a (786 mg, 2.24 mmol) and triphenylphosphine (978 mg, 3.73 mmol) were added to THF (10 mL), DIAD was added thereto at 0° C. (730 μL, 3.75 mmol), and the reaction was allowed to run at room temperature for 15 hours. After the completion of the reaction, the solvent was removed under reduced pressure, and the resulting residue was directly used in the next step. LCMS: [M+Na]=656.2.

(75) Step 9: Synthesis of Compound WX011-9

(76) Compound WX011-8 (0.9 g, 1.42 mmol) was dissolved in benzyl alcohol (15 mL), a solution of hydrogen chloride in 1,4-dioxane (4 M, 15 mL) was added thereto, and the reaction was allowed to run at 50° C. for 1 hour. After the completion of the reaction, the solvent was removed under reduced pressure, and the resulting residue was slurried with methyl tert-butyl ether (150 mL) to give the target compound WX011-9. LCMS: [M+H].sup.+=643.4.

(77) Step 10: Synthesis of Compound WX011-10

(78) DCM (0.7 mL) was added to compound WX011-9 (0.1 g, 155.58 μmol), sodium hydroxide (1 M, 0.8 mL) was added, followed by acetyl chloride (44 μL, 622 μmol), and then the reaction was allowed to run at room temperature for 1 hour. The reaction mixture was extracted with DCM (5 mL×2). The combined organic phase was dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the solvent was removed from the filtrate under reduced pressure. The resulting residue was separated by preparative chromatography (hydrochloric acid condition) to give the target compound WX011-10. LCMS: [M+H].sup.+=707.1.

(79) Step 11: Synthesis of Compound WX011

(80) Compound WX011-10 (50 mg, 73.01 μmol) was dissolved in MeOH (5 mL), 10% Pd/C (100 mg) was added under nitrogen atmosphere. The reaction system was purged with hydrogen for 3 times, and then the reaction was allowed to run under hydrogen atmosphere (15 Psi) at room temperature for 1 hour. The reaction mixture was filtered, and the solvent was removed under reduced pressure to give a residue. The residue was separated by preparative chromatography (hydrochloric acid condition) to give the target compound WX011.

(81) Compound WX011 was analyzed by supercritical fluid chromatography (column: (S,S)Whelk-01, 100×4.6 mm 5 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in methanol; gradient: B from 5% to 40% in 4.5 minutes, 40% for 2.5 minutes, back to 5% equilibrium for 1 minute; flow rate: 2.8 mL/min; column temperature: 40° C.; wavelength: 220 nm) as racemates. Chiral isomers WX011A and WX011B were separated, and their retention times were 3.078 min and 3.734 min, respectively.

(82) WX011A, .sup.1HNMR (400 MHz, CDCl.sub.3) δ 7.70 (s, 1H), 7.47-7.42 (m, 1H), 7.30-7.26 (m, 2H), 6.99-6.96 (m, 1H), 6.86-6.84 (m, 1H), 5.47-5.42 (m, 1H), 4.39-4.30 (m, 1H), 4.15-4.10 (m, 1H), 3.82 (s, 3H), 3.47-3.00 (m, 5H), 2.95 (s, 3H), 1.93, 1.91 (2 s, 3H), 1.54-1.19 (m, 10H); LCMS (5-95 AB/1.5 min): Rt=0.904; [M+Na]=617.3.

(83) WX011B, .sup.1HNMR (400 MHz, CDCl.sub.3) δ 7.70 (s, 1H) 7.47-7.42 (m, 1H) 7.30-7.26 (m, 2H) 6.99-6.96 (m, 1H) 6.86-6.84 (m, 1H) 5.47-5.42 (m, 1H) 4.39-4.33 (m, 1H) 4.15-4.10 (m, 1H) 3.82 (s, 3H) 3.47-3.00 (m, 5H) 2.95 (s, 3H) 1.93, 1.91 (2 s, 3H) 1.54-1.19 (m, 10H); LCMS (5-95 AB/1.5 min): Rt=0.904; [M+Na]=617.4.

(84) Referring to the synthetic methods of steps 10 and 11 in the example 1, each example in the following table was synthesized using different intermediate fragments in step 10. The structures in the table also represent their possible isomers.

(85) TABLE-US-00003 TABLE 3 Compound structure of each example Intermediate Example fragment Structure Compound 12 WX012 WX012A or WX012B WX012B or WX012A 13 MeI WX013 14 0 WX014 15 WX015 WX015A or WX015B WX015B or WX015A

(86) TABLE-US-00004 TABLE 4 NMR and MS data of each example Example Compound NMR MS m/z: 12 SFC detection method: Column: (S,S)Whelk-01, 100 × 4 6 mm I.D., 5 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in methanol; gradient: B from 5% to 40% in 4.5 minutes, 40% for 2.5 minutes, back to 5% equilibrium for 1 minute; flow rate: 2.5 mL/min; column temperature: 40° C.; wavelength: 220 nm. WX012 A pair of racemates with a ratio of 1:1, detected by SFC. 631.1 WX012A SFC retention time 2.649 min (M + H).sup.+ .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.70 (s, 1H) 7.41-7.39 (m, 1H) 7.28-7.26 (m, 2H) 6.99-6.95 (m, 1H) 6.87- 7.85 (m, 1H) 5.43-5.41 (m, 1H) 4.40-4.37 (m, 1 H) 4.11-4.06 (m, 1H) 3.83 (s, 3H) 3.45-3.41 (m, 1 H) 3.15-3.00 (m, 1H) 2.90 (s, 1H) 2.85-2.72 (m, 1 H) 2.39 (s, 3H) 2.30-2.27 (m, 1H) 1.66-1.18 (m, 10 H). WX012B SFC retention time 3.874 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.70 (s, 1H) 7.41-7.39 (m, 1 H) 7.28-7.26 (m, 2H) 6.99-6.95 (m, 1H) 6.87- 7.85 (m, 1H) 5.43-5.41 (m, 1H) 4.40-4.37 (m, 1 H) 4.11-4.06 (m, 1H) 3.83 (s, 3H) 3.45-3.41 (m, 1 H) 3.15-3.00 (m, 1H) 2.90 (s, 1H) 2.85-2.72 (m, 1 H) 2.39 (s, 3H) 2.30-2.27 (m, 1H) 1.66-1.18 (m, 10 H). 13 WX013 .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.84-7.82 (m, 1H), 7.45- 567.4 7.30 (m, 3H), 7.10-7.09 (m, 1H), 7.0-6.98 (m, 1 (M + H).sup.+ H), 5.60-5.52 (m, 1H) 4.53-4.49 (m, 1H), 4.25- 4.22 (m, 1H), 3.98-3.96 (m, 4H), 3.65-3.65 (m, 1 H), 3.03-2.99 (m, 5H), 2.65-2.35 (m, 4H), 1.70- 1.50 (m, 10H). 14 WX014 .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.70 (s, 1H) 7.44-7.43 653.1 (m, 1H) 7.26-7.20 (m, 2H) 6.99-6.95 (m, 1H) 6.84- (M + H).sup.+ 7.82 (m, 1H) 5.42-5.41 (m, 1H) 4.38-4.34 (m, 1 H) 4.18-4.12 (m, 1H) 3.80 (s, 3H) 3.45-3.42 (m, 1 H) 3.29-3.19 (m, 1H) 2.96 (s, 3 H) 2.95-2.85 (m, 2 H) 1.52 (s, 3H) 1.50 (s, 3H) 1.35-1.31 (m, 3H) 1.18 (s, 9H) 1.04-1.03 (m, 1H). 15 SFC detection method: column: Chiralcel OJ-3, 100 × 4.6 mm I.D., 3 μm; mobile phase: A: supercritical carbon dioxide, B: 0.05% diethylamine in ethanol; gradient: B from 5% to 40% in 4.5 minutes, 40% for 2.5 minutes, back to 5% equilibrium for 1 minute; flow rate: 2.5 mL/min; column temperature: 40° C.; wavelength: 220 nm. WX015 The crude product obtained after hydrogenation was 611.4 directly separated by SFC to give the enantiomers. As (M + H).sup.+ racemates with a ratio of 1:1, detected by SFC. WX015A SFC retention time 2.475 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.70 (s, 1H) 7.44 (d, J = 7.2 Hz, 1H) 7.28-7.20 (m, 2H) 7.00-6.95 (m, 1H) 6.85-7.83 (m, 1H) 5.43-5.41 (m, 1H) 4.38-4.34 (m, 1H) 4.16-4.11 (m, 1H) 3.80 (s, 3H) 3.55 (s, 3H) 3.45- 3.05 (m, 5H) 2.96 (s, 1H) 1.60-1.10 (m, 10H). WX015B SFC retention time 2.658 min .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.70 (s, 1H) 7.44 (d, J = 7.2 Hz, 1H) 7.28-7.20 (m, 2H) 7.00-6.95 (m, 1H) 6.85-7.83 (m, 1H) 5.43-5.41 (m, 1H) 4.38-4.34 (m, 1H) 4.16-4.11 (m, 1H) 3.80 (s, 3H) 3.55 (s, 3H) 3.45- 3.05 (m, 5H) 2.96 (s, 1H) 1.60-1.10 (m, 10H).

Experimental Example 1: In Vitro Evaluation

(87) Objective of this Experiment:

(88) The IC.sub.50 value was determined to evaluate the ability of the test compounds to inhibit acetyl-CoA carboxylase (ACC).

(89) Experimental Materials:

(90) Protein: human acetyl-CoA carboxylase 1 (hACC1) and human acetyl-CoA carboxylase 2 (hACC2).

(91) Substrate: NaHCO.sub.3

(92) Cofactor: acetyl coenzyme A, ATP

(93) Activator: potassium citrate

(94) Experimental Method:

(95) 1. 1 time of the enzyme/substrate/cofactor was added to the wells of the well plate.

(96) 2. According to Acoustic technology, the solutions of the compounds in DMSO were added to the above enzyme mixture and the mixture was pre-incubated for 15 minutes.

(97) 3. ATP was added to initiate the reaction and the mixture was shaken until thoroughly mixed.

(98) 4. The mixture was incubated for 1 hour at room temperature.

(99) 5. After quenching the reaction, the incubation continued for 40 minutes.

(100) 6. After adding the detection reagents, the mixture was incubated for 30 minutes.

(101) 7. Measure Luminescence.

(102) 8. Data analysis: based on the standard curve of ADP, the luminescence was converted into ADP product concentration and the enzyme activity was calculated. Graphpad Prism software was used to fit the curve to obtain the IC.sub.50 value. The experimental results are shown in Table 5.

(103) TABLE-US-00005 TABLE 5 In vitro screening test results of the compounds of the present disclosure Compound hACC1 (nM) hACC2(nM) WX001B 4.9 8 WX002 53.6 8.7 WX003B 10.9 3.4 WX004B 14.1 10.3 WX005 46.6 45 WX006 20.3 22.1 WX007B 9.1 12.6 WX008B 5.3 13.9 WX009B 5.6 11 WX010 83.9 70.7 WX011 26 10.9 WX012 14.2 5.7 WX014 26 7.2 WX015 20.2 7.0 WX016B 11.1 4.8

(104) Conclusion: The compounds of the present disclosure have a strong inhibitory activity on human ACC1/ACC2 enzyme.

Experimental Example 2: Evaluation of the Pharmacokinetics Characteristics of the Compounds

(105) Objective of this Experiment:

(106) Testing the pharmacokinetics characteristics of the compounds in C57BL/6 mice

(107) Experimental Materials:

(108) C57BL/6 mice (male, 18-30 g, 7-9 weeks, Shanghai Lingchang Biotechnology Co., Ltd.)

(109) Experimental Operation:

(110) The clear solution of the test compounds (0.5 mg/mL in 10% DMSO, 10% polyethylene glycol stearate, 80% water) was injected via tail vein into 4 male C57BL/6 mice (overnight fasted, 7-9 weeks), at a dose of 2.0 mg/kg. The suspensions or clear solutions of the test compounds (1 mg/mL in 10% PEG400, 90% (0.5% methylcellulose+0.2% Tween 80)) were administrated by oral gavage to 4 male C57BL/6 mice (overnight fasted, 7-9 weeks), at a dose of 10 mg/kg.

(111) Two mice were set in each group and subjected to blood sampling alternatively, 4 to 5 time points per mouse. At 0.0833h (IV group only), 0.25 h, 0.5 h, 1.0 h, 2.0 h, 4.0 h, 6.0 h, 8.0 h, and 24 h after the intravenous or oral administration, about 30 μL of blood was collected by saphenous vein puncture and added into an anticoagulation tube with EDTA-K.sub.2, and the plasma was isolated by centrifugation. The drug plasma concentration was determined by LC-MS/MS, and WinNonlin™ Version 6.3 (Pharsight, Mountain View, Calif.) pharmacokinetic software was used to calculate relevant pharmacokinetic parameters using the non-compartmental model linear logarithmic trapezoidal method.

(112) The experimental results are shown in Table 6:

(113) TABLE-US-00006 TABLE 6 Pharmacokinetic test results The test sample (compounds Bioavail- prepared in Clearance Half-life AUC.sub.0-last ability each example) (mL/min/kg) T.sub.1/2 (h) (nM .Math. h) F (%) WX004B 27.3 1.85 7211 66.7 WX008B 51.9 0.60 304 5.86 WX009B 92.1 0.61 40 1.26

(114) Conclusion: The compounds of the present disclosure can significantly improve single or partial pharmacokinetic parameters in mouse.

Experimental Example 3: In Vivo Pharmacodynamic Study in NASH Mouse Model Induced by HFD+CCl.SUB.4

(115) Objective of this Experiment:

(116) The objective of this research is to study the effect of the compound on improving NASH and liver fibrosis in HFD+CCl.sub.4 mouse model, with 1-181 as the reference compound.

(117) 1-181 is an Acetyl-CoA carboxylase inhibitor and is currently undergoing a phase II clinical study on non-alcoholic fatty liver disease (NAFLD). The HFD+CCl.sub.4 mouse model used in this study is an animal model simulating human non-alcoholic fatty liver disease evolving into NASH, high-fat diet causes the fat accumulation and steatosis in liver cells; CCl.sub.4 (intraperitoneal injection, twice a week) simulates the “second hit” of liver injury. This model is stable and reliable and has a high similarity to the pathogenesis of human NASH, it has the main pathological characteristics of NASH, including steatosis, apoptosis, inflammation and fibrosis, and also shows elevated plasma aminotransferase (ALT and AST) levels.

(118) Experimental Design:

(119) The modeling for this experiment included two steps of high-fat feed and CCl.sub.4 induction. Firstly, the mice were fed with high-fat feed to induce non-alcoholic fatty liver, and mice with body weight >38 g were selected. The mice were continuously fed with high-fat feed, and simultaneously intraperitoneally injected with 25% CCl.sub.4, 0.5 mg/kg, twice a week, for a total of four weeks. The day of starting CCl.sub.4 administration was set as day 0, and the time of starting CCl.sub.4 administration was set as hour 0. On the day of starting CCl.sub.4 administration, the intragastric administration was started, and the administration volume of each group was 5 mL/kg, once a day for 4 weeks (28 days). The injection time of CCl.sub.4 should be more than 4 hours away from the first drug administration time point in this day. 6 groups were set in this experiment, namely the healthy control group, model group, reference compound group (GS-0976), test compound group (WX004B, three doses). The healthy control group had 10 normal mice, which were fed with normal feed during the experiment, without CCl.sub.4 injection; 50 obese mice were used in the model group and the administration group, 10 mice each group. After grouping, CCl.sub.4 was injected intraperitoneally and the different doses of the drugs were administered. The grouping and dosage regimen are shown in Table 7.

(120) TABLE-US-00007 TABLE 7 Animal grouping and dosing regimen Feed Dosing regimen (dose | mode of and Number of Test administration | frequency | total CCl.sub.4 Group animals compound Menstruum duration) injection Healthy 10 Menstruum 40% 0| Oral gavage| QD | Day 0-27 Normal control polyethylene feed, group glycol/10% without Solutol/50% CCl.sub.4 water injection Model 10 Menstruum 40% 0| Oral gavage| QD |Day 0-27 High- group polyethylene fat feed, glycol/10% CCl.sub.4 Solutol/50% injection water I-181, 3 10 I-181 40% 3 mg/kg| Oral gavage| QD | High- mpk polyethylene Day 0-27 fat feed, glycol/10% CCl.sub.4 Solutol/50% injection water WX004 10 WX004B 40% 0.5 mg/kg| Oral gavage| QD | High- B, 0.5 polyethylene Day 0-27 fat feed, mpk glycol/10% CCl.sub.4 Solutol/50% injection water WX004 10 WX004B 40% 1 mg/kg| Oral gavage| QD | High- B, 1 polyethylene Day 0-27 fat feed, mpk glycol/10% CCl.sub.4 Solutol/50% injection water WX004 10 WX004B 40% 3 mg/kg| Oral gavage| QD | High- B, 3 polyethylene Day 0-27 fat feed, mpk glycol/10% CCl.sub.4 Solutol/50% injection water

(121) Experimental Results:

(122) In the mouse model induced by the combination of a high-fat diet and CCl.sub.4, WX004B achieves the same efficacy in both NAS and fibrosis as the reference compound at a higher dose.