Pyrazolopyrimidine derivative and use thereof

Abstract

A compound of formula (II), a tautomer thereof or a pharmaceutically acceptable salt thereof, and use thereof in the preparation of medicaments for treating solid tumor-related diseases. ##STR00001##

Claims

1. A compound represented by formula (II), an isomer thereof or a pharmaceutically acceptable salt thereof, ##STR00069## wherein, T.sub.1, T.sub.2, T.sub.3, T.sub.4, T.sub.5, and T.sub.6 are each independently selected from the group consisting of CR.sub.3 and N; W is selected from the group consisting of CR.sub.4 and N; X.sub.1 and X.sub.2 are each independently CR.sub.5R.sub.6; R.sub.1 is selected from the group consisting of H, F, Cl, Br, I, OH, NH.sub.2, CN and C.sub.1-6 alkyl, wherein the C.sub.1-6 alkyl is optionally substituted by 1, 2 or 3 R.sub.a; R.sub.2 is selected from the group consisting of H 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 and R.sub.4 are each independently selected from the group consisting of H, F, Cl, Br, I, OH and NH.sub.2; R.sub.5 and R.sub.6 are each independently selected from the group consisting of H, F, Cl, Br, I, OH, NH.sub.2 and C.sub.1-6 alkyl; L.sub.1 is selected from the group consisting of —C.sub.1-3 alkyl—, —C.sub.3-6 cycloalkyl— and -4-to 6-membered heterocycloalkyl—, wherein the —C.sub.1-3 alkyl—, —C.sub.3-6 cycloalkyl— and -4- to 6-membered heterocycloalkyl— are optionally substituted by 1, 2 or 3 R.sub.c; L.sub.2 is selected from the group consisting of —C.sub.1-3 alkyl—, —C.sub.1-3 alkyl—O—, —N(Rd)—, —C.sub.1-3 alkyl—N(Rd)— and —O—; R.sub.a is independently selected from the group consisting of H, F, Cl, Br, I, OH and NH.sub.2; R.sub.b is selected from the group consisting of H, F, Cl, Br, I, OH and NH.sub.2; R.sub.c is selected from the group consisting of H, F, Cl, Br, I, OH, NH.sub.2, CN, C.sub.1-3 alkyl and C.sub.1-3 alkyl—C═O—, wherein the C.sub.1-3 alkyl and C.sub.1-3 alkyl—C═O— are optionally substituted by 1, 2 or 3 R; R.sub.d is selected from the group consisting of H and C.sub.1-3 alkyl; R is independently selected from the group consisting of F, Cl, Br, I, OH and NH.sub.2; the carbon atom marked with “*” is a chiral carbon atom present in a single enantiomer form of (R) or (S) or in a form enriched in one enantiomer; the 4- to 6-membered heterocycloalkyl independently comprises 1, 2, 3 or 4 heteroatoms or heteroatomic groups independently selected from the group consisting of —NH—, —O—, —S—and N.

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

3. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 1, wherein R.sub.2 is selected from the group consisting of H and CH.sub.3.

4. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 1, wherein R.sub.5 and R.sub.6 are each independently selected from the group consisting of H, F, Cl, Br, I, OH, NH.sub.2 and CH.sub.3.

5. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 1, wherein R.sub.c is selected from the group consisting of H, F, Cl, Br, I, OH, NH.sub.2, CN, CH.sub.3, CH.sub.3 CH.sub.2 and CH.sub.3 C(=O), wherein the CH.sub.3, CH.sub.3 CH.sub.2 and CH.sub.3 C(=O) are optionally substituted by 1, 2 or 3 R.

6. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 5, wherein R.sub.c is selected from the group consisting of H, F, Cl, Br, I, OH, NH.sub.2, CN, CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, CH.sub.3 CH.sub.2 and CH.sub.3 C(=O).

7. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 1, wherein L.sub.1 is selected from the group consisting of —CH.sub.2—, —CH.sub.2CH.sub.2—, —cyclopropyl—, —cyclobutyl—, —cyclopentyl—, —oxetanyl—, —tetrahydrofuranyl—, —tetrahydropyranyl—, —pyrrolidinyl— and —piperidinyl—, wherein the —CH.sub.2—,—CH.sub.2CH.sub.2—, —cyclopropyl—, —cyclobutyl—, —cyclopentyl—, —oxetanyl—, — tetrahydrofuranyl—, —tetrahydropyranyl—, —pyrrolidinyl— and —piperidinyl— are optionally substituted by 1, 2 or 3 R.sub.c.

8. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 7, wherein L.sub.1 is selected from the group consisting of —CH.sub.2—, —CH(CH.sub.3)—, —C(CH.sub.3).sub.2—, —CH.sub.2CH.sub.2—, ##STR00070##

9. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 8, wherein L.sub.1 is selected from the group consisting of —CH.sub.2—, —CH.sub.2CH.sub.2—, —CH(CH.sub.3)—, —C(CH.sub.3).sub.2—, ##STR00071##

10. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 1, wherein L.sub.2 is selected from the group consisting of —CH.sub.2—, —CH.sub.2CH.sub.2—, —CH.sub.2CH.sub.2O—, —CH(CH.sub.3)O—, —O—, —NH—, —CH.sub.2NH— and —CH.sub.2O—.

11. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 1, wherein the moiety ##STR00072## is selected from the group consisting of ##STR00073##

12. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 1, wherein the moiety ##STR00074##

13. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 1, wherein the moiety ##STR00075##

14. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 1, wherein the compound is selected from: ##STR00076## wherein, W, R.sub.1, R.sub.2, L.sub.1 and L.sub.2 are as defined in claim 1.

15. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 14, wherein the compound is selected from: ##STR00077## wherein, n is selected from the group consisting of 0 and 1.

16. A compound, an isomer thereof or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: ##STR00078## ##STR00079## ##STR00080##

17. The compound, isomer thereof or pharmaceutically acceptable salt thereof as defined in claim 16, wherein the compound is selected from ##STR00081## ##STR00082## ##STR00083## ##STR00084## ##STR00085## ##STR00086##

18. A pharmaceutical composition comprising a therapeutically effective amount of the compound or pharmaceutically acceptable salt thereof as defined in claim 1 as active ingredient and a pharmaceutically acceptable carrier.

19. A method for treating a disease related to Trk, ALK and Ros1 kinase in a subject in need thereof, comprising: administering an effective amount of the compound or pharmaceutically acceptable salt thereof as defined in claim 1 to the subject, the disease related to Trk, ALK and Ros1 kinase is a colon cancer or a lung cancer.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1: transplanted tumor model of human colon cancer KM12 in nude mice;

(2) FIG. 2: subcutaneous xenograft tumor model of human lung cancer LU-01-0414;

(3) PO stands for oral administration; QD stands for once a day; BID stands for twice a day.

DETAILED DESCRIPTION OF THE EMBODIMENT

(4) The following examples further illustrate the present disclosure, but the present disclosure is not limited thereto. The present disclosure has been described in detail in the text, and its specific embodiments have also been disclosed, for one skilled in the art, it is obvious to modify and improve the embodiments of the present disclosure within the spirit and scope of the present disclosure.

Example 1: Synthesis of WX001

(5) ##STR00033## ##STR00034## ##STR00035##

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

(7) Compound 1-1 (15 g, 96.70 mmol, 1 eq) was dissolved in ethyl acetate (300 mL), followed by addition of isopropylidene malonate (13.94 g, 96.70 mmol, 1 eq), triethylene diamine (1.08 g, 9.67 mmol, 1.06 mL, 0.1 eq) and tert-butyl N-hydroxycarbamate (12.87 g, 96.70 mmol, 1 eq). The obtained reaction mixture was stirred at 25° C. for 16 hours. The reaction mixture was washed twice with water (200 mL each time), and then washed once with 100 mL saturated brine. The organic phase was dried over anhydrous sodium sulfate, the desiccant was removed by filtration, and the solvent was removed from the filtrate under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether:ethyl acetate=3:1) to obtain compound 1-2. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 7.98 (d, J=2.8 Hz, 1H), 7.52-7.48 (m, 1H), 5.65 (dd, J=3.6, 10.0 Hz, 1H), 3.95 (s, 3H), 3.33 (dd, J=9.6, 18.0 Hz, 1H), 2.74 (dd, J=3.6, 16.0 Hz, 1H), 1.50 (s, 9H). LCMS m/z=313.3 [M+H].sup.+.

(8) Step 2: Synthesis of Compound 1-3

(9) Compound 1-2 (21.40 g, 68.53 mmol, 1 eq) was dissolved in tetrahydrofuran (300 mL), and lithium borohydride (4.48 g, 205.58 mmol, 3 eq) was slowly added, and stirred at 25° C. for 0.1 hour. 200 mL of water was added to the reaction mixture, and then extracted twice with ethyl acetate (50 mL each time). The organic phases were combined and washed with 100 mL of saturated brine, then dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated to dryness by rotary evaporation to obtain the crude compound 1-3. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 7.88 (d, J=3.2 Hz, 1H), 7.67-7.64 (dd, J=2.8, 8.8 Hz, 1H), 5.46-5.42 (m, 1H), 3.91 (s, 3H), 3.86-3.71 (m, 2H), 2.24-2.14 (m, 1H), 2.08-2.00 (m, 1H), 1.41 (s, 9H). LCMS m/z=317.3[M+H].sup.+.

(10) Step 3: Synthesis of Compound 1-4

(11) Compound 1-3 (14.52 g, 45.90 mmol, 1 eq) and triphenylphosphine (30.10 g, 114.76 mmol, 2.5 eq) were dissolved in tetrahydrofuran (150 mL), and the obtained reaction mixture was cooled to 5° C. in an ice-water bath, followed by dropwise addition of diisopropyl azodicarboxylate (27.85 g, 137.71 mmol, 26.77 mL, 3 eq). After the dropwise addition, the ice-water bath was removed, and the mixture was stirred at 25° C. for 0.1 hour. The reaction mixture was concentrated to dryness by rotary evaporation, and the residue was purified by column chromatography (petroleum ether:ethyl acetate=50:1 to 30:1 to 10:1 to 5:1) to obtain compound 1-4. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 7.88 (d, J=3.2 Hz, 1H), 7.52-7.50 (m, 1H), 5.38-5.35 (m, 1H), 4.13-4.03 (m, 1H), 3.94 (s, 3H), 3.89-3.82 (m, 1H), 2.84-2.76 (m, 1H), 2.12-2.03 (m, 1H), 1.50 (s, 9H). LCMS m/z=299.3[M+H].sup.+.

(12) Step 4: Synthesis of Compound 1-5

(13) Compound 1-4 (3.00 g, 10.06 mmol, 1 eq) was dissolved in a solution of hydrogen chloride in methanol (4 M, 12.57 mL, 5 eq) and stirred at 25° C. for 3 hours. The reaction mixture was concentrated to dryness by rotary evaporation and compound 1-5 was obtained. .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.17 (d, J=2.8 Hz, 1H), 7.81-7.79 (m, 1H), 5.21 (t, J=8.0 Hz, 1H), 4.60-4.54 (m, 1H), 4.40-4.32 (m, 1H), 4.04 (s, 3H), 2.96-2.80 (m, 2H). LCMS m/z=199.3 [M+H].sup.+.

(14) Step 5: Synthesis of Compound 1-6

(15) Ethyl 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate (1.92 g, 8.52 mmol, 1 eq), compound 1-5 (2.20 g, 9.38 mmol, 1.1 eq) and n-butanol (5 mL) were added into a reaction flask, and then N,N-diisopropylethylamine (6.61 g, 51.14 mmol, 8.91 mL, 6 eq) was added, and the obtained reaction mixture was stirred at 90° C. for 3.5 hours. The reaction mixture was concentrated, 30 mL of water was added, and then extracted with 30 mL of ethyl acetate. The organic phase was separated, washed once with 20 mL of saturated brine, then dried over anhydrous sodium sulfate, and filtered to remove the desiccant. The solvent was removed from the filtrate under reduced pressure to obtain a crude product, which was purified by column chromatography (petroleum ether:ethyl acetate=100:0 to 10:1 to 5:1 to 2:3) to obtain compound 1-6. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 8.48 (d, J=7.6 Hz, 1H), 8.39 (s, 1H), 7.92 (d, J=3.2 Hz, 1H), 7.58-7.55 (m, 1H), 7.03 (d, J=7.6 Hz, 1H), 6.06 (dd, J 5.2, 8.8 Hz, 1H), 4.33-4.24 (m, 2H), 4.22-4.18 (m, 1H), 4.01 (s, 3H), 3.93-3.87 (m, 1H), 2.94-2.90 (m, 1H), 2.36-2.30 (m, 1H), 1.27 (t, J=6.8 Hz, 3H). LCMS m/z=388.3 [M+H].sup.+.

(16) Step 6: Synthesis of Compounds 1-7

(17) Compound 1-6 (3.9 g, 10.07 mmol, 1 eq) was dissolved in acetonitrile (100 mL), sodium iodide (4.53 g, 30.20 mmol, 3 eq) was added, and trimethylchlorosilane (3.28 g, 30.20 mmol, 3.83 mL, 3 eq) was added dropwise under stirring. After the dropwise addition, the obtained reaction mixture was stirred and refluxed at 75° C. for 0.5 hour under nitrogen atmosphere. 50 mL of water was added to the reaction mixture and then a solid precipitated. The mixture was filtered, and the filter cake was dried under vacuum at 40° C. to obtain compound 1-7. .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.69 (d, J=7.6 Hz, 1H), 8.30 (s, 1H), 7.63-7.60 (m, 1H), 7.38 (t, J=3.2 Hz, 1H), 7.10 (d, J 5.6, 1H), 5.79-5.75 (m, 1H), 4.26-4.19 (m, 2H), 2.98-2.90 (m, 1H), 2.35-2.29 (m, 1H), 1.25 (t, J=7.2 Hz, 3H). LCMS m/z=374.3 [M+H].sup.+.

(18) Step 7: Synthesis of Compounds 1-8

(19) Compound 1-7 (0.6 g, 1.61 mmol, 1 eq) and triethylamine (442.34 mg, 4.37 mmol, 608.45 μL, 2.72 eq) were dissolved in anhydrous dichloromethane (20 mL), and cooled to 5° C. in an ice-water bath, followed by dropwise addition of trifluoromethanesulfonic anhydride (1.22 g, 4.31 mmol, 710.64 μL, 2.68 eq). After the dropwise addition, the obtained reaction mixture was naturally warmed to 25° C. and stirred for 2 hours under nitrogen atmosphere. The reaction mixture was washed with 20 mL of water and 15 mL of saturated brine, then dried over anhydrous sodium sulfate, and filtered to remove the desiccant. The filtrate was concentrated to dryness by rotary evaporation to obtain compound 1-8. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 8.53 (d, J=7.6 Hz, 1H), 8.43 (s, 1H), 8.14 (d, J=2.8 Hz, 1H), 7.87 (dd, J=2.8, 7.6 Hz, 1H), 7.05 (d, J=7.2 Hz, 1H), 6.10 (dd, J=5.6, 8.8 Hz, 1H), 4.37-4.24 (m, 3H), 3.95-3.88 (m, 1H), 3.12-3.04 (m, 1H), 2.50-2.41 (m, 1H), 1.28 (t, J=7.2 Hz, 3H). LCMS m/z=506.3 [M+H].sup.+.

(20) Step 8: Synthesis of Compounds 1-9

(21) Compound 1-8 (3.00 g, 5.94 mmol, 1 eq) was dissolved in a mixture of water (60 mL) and toluene (120 mL), diisopropylamine (1.50 g, 14.84 mmol, 2.10 mL, 2.5 eq), bis(triphenylphosphine)palladium dichloride (833.28 mg, 1.19 mmol, 0.2 eq) and cuprous iodide (226.10 mg, 1.19 mmol, 0.2 eq) were added, and (R)—N-Boc-3-amino-1-butyne (4.02 g, 23.74 mmol, 4 eq) was added at last. The obtained reaction mixture was allowed to react at 100° C. for 16 hours under nitrogen atmosphere. The reaction mixture was filtered, and the filter cake was washed with 20 mL of ethyl acetate. The filtrate was separated to obtain an organic phase, which was dried over anhydrous sodium sulfate and then filtered to remove the desiccant. The filtrate was concentrated to dryness by rotary evaporation to obtain a crude product, which was purified by column chromatography (petroleum ether:ethyl acetate=10:1 to 5:1 to 1:1) to obtain compound 1-9. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 8.52 (d J=7.6 Hz, 1H), 8.38 (d, J=3.6 Hz 1H), 8.36-8.33 (m, 1H), 7.65-7.62 (m, 1H), 7.05 (t, J=7.6 Hz, 1H), 6.44-6.38 (m, 1H), 5.76-5.54 (brs, 1H), 4.91-4.76 (m, 1H), 4.36-4.24 (m, 2H), 3.93-3.83 (m, 1H), 3.08-3.04 (m, 1H), 2.41-2.29 (m, 1H), 1.60-1.54 (m, 3H), 1.43 (s, 9H), 1.32-1.29 (m 3H). LCMS m/z=525.5[M+H].sup.+.

(22) Step 9: Synthesis of Compound 1-10

(23) Compound 1-9 (1.1 g, 2.10 mmol, 1 eq) was dissolved in ethanol (20 mL), and palladium/carbon (2.10 mmol, 10% purity, 1 eq) and sodium carbonate (444 mg, 4.19 mmol, 2 eq) were added. The reaction mixture was purged with hydrogen, and then stirred at 25° C. for 1.5 hours under hydrogen atmosphere at a pressure of 15 psi. The reaction mixture was filtered, and the filtrate was concentrated to dryness by rotary evaporation to obtain 712 mg of crude product. The crude product was purified by preparative plate (petroleum ether:ethyl acetate=1:1.5) to obtain compound 1-10, which was directly used in the reaction of next step. LCMS m/z=529.5 [M+H].sup.+.

(24) Step 10: Synthesis of Compound 1-11

(25) Compound 1-10 (10 mg, 18.92 μmol, 1 eq) was dissolved in methanol (1 mL), and then a prepared sodium hydroxide solution (3 M, 37.84 μL, 6 eq) and water (0.04 mL) were added. The resulting reaction mixture was stirred at 60° C. for 1.5 hours. The above reaction was conducted in octuplicate, and the 8 batches of reaction mixture were combined together, neutralized with 1 mol/L dilute hydrochloric acid until the pH value was 7, and then concentrated to dryness by rotary evaporation to obtain a crude product. The crude product was purified by high performance preparative chromatography to obtain compound 1-11. LCMS m/z=501.2 [M+H].sup.+, 401.4[M−100+H].sup.+.

(26) Step 11: Synthesis of Compound 1-12

(27) Compound 1-11 (8.6 mg, 17.18 μmol, 1 eq) was dissolved in a solution of hydrogen chloride in ethyl acetate (3 M, 0.6 mL, 104.76 eq), and stirred at 20° C. for 1 hour. The reaction was conducted in duplicate, and the 2 batches of reaction mixture were combined, and concentrated to dryness by rotary evaporation to obtain a crude product of compound 1-12, which was directly used in the reaction of next step. LCMS m/z=401.3 [M+H].sup.+.

(28) Step 12: Synthesis of Compound WX001

(29) Compound 1-12 (13.8 mg, 34.47 μmol, 1 eq) was dissolved in N,N-dimethylformamide (5 mL), followed by addition of pentafluorophenyl diphenylphosphate (19.86 mg, 51.70 μmol, 1.5 eq) and then N,N-diisopropylethylamine (11.14 mg, 86.16 μmol, 15.01 μL, 2.5 eq). The reaction mixture was stirred at 25° C. for 1 hour. 30 mL of dichloromethane was added to the reaction mixture, and then washed with water (10 mL×3). The organic phase was concentrated to dryness by rotary evaporation and the residue was dissolved in 130 mL of methyl tert-butyl ether, and then washed with water (10 mL×3). The organic phase was concentrated to obtain a crude product. The crude product was separated by HPLC (hydrochloric acid system) to obtain a hydrochloride of compound WX001. .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.83 (s, 1H), 8.79 (d, J=7.2 Hz, 1H), 8.29 (s, 1H), 7.05 (d, J=7.2 Hz, 1H), 6.24-6.21 (m, 1H), 4.63 (t, J=7.2 Hz, 1H), 4.30-4.25 (m, 1H), 3.96-4.03 (m, 1H), 3.84-3.78 (m, 1H), 3.35-3.30 (m, 2H), 3.20-3.13 (m, 1H), 2.50-2.69 (m, 2H), 2.04-1.99 (m, 1H), 1.44 (d, J=6.4 Hz, 3H). LCMS m/z=385.2 [M+H].sup.+.

(30) The hydrochloride of compound 001 was dissolved in methanol, and a basic resin (model: Amberlite IRA-400) was added under stirring. After 0.5 hour, the mixture was basic determined by pH test, and was filtered to remove the resin and directly concentrated to dryness to obtain compound WX001.

Example 2-4: Synthesis of WX002, WX002A and WX002B

(31) ##STR00036## ##STR00037##

(32) Steps 1-6: Compounds 1-2 to 1-7 were Synthesized by Steps Similar to the Step 1 to Step 6 in Example 1

(33) Step 7: Synthesis of Compound 2-8

(34) Compound 1-7 was dissolved in methanol (30 mL), and a prepared solution of sodium hydroxide (385.68 mg, 9.64 mmol, 4 eq) in water (3 mL) was added. The resulting reaction mixture was stirred at 60° C. under nitrogen atmosphere for 16 hours. The reaction mixture was cooled to room temperature, and the pH value was adjusted to about 7 with 2M hydrochloric acid solution. The mixture was directly concentrated to dryness by rotary evaporation to obtain compound 2-8, which was directly used in the next step. LCMS m/z=346.2 [M+H].sup.+.

(35) Step 8: Synthesis of Compound 2-9

(36) Compound 2-8 was dissolved in N,N-dimethylformamide (8 mL), then N,N-diisopropylethylamine (449.36 mg, 3.48 mmol, 605.60 μL, 3.5 eq) and O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (453.26 mg, 1.19 mmol, 1.2 eq) were added and stirred for 0.5 hour, and then (1-(hydroxymethyl)cyclopropylamino hydrochloride (159.59 mg, 1.29 mmol, 1.3 eq, HCl) was added. The resulting reaction mixture was allowed to react at 25° C. for 3 hours. The reaction mixture was poured into 80 mL of saturated aqueous solution of ammonium chloride, then extracted with dichloromethane (60 mL×3). The organic phases were combined and washed with saturated brine (60 mL×3), dried over anhydrous sodium sulfate in an appropriate amount, filtered to remove the desiccant, and the filtrate was concentrated to obtain a crude product. 2 mL of water was added to the crude product, and then freeze-dried to obtain compound 2-9, which was directly used in the reaction of next step. LCMS m/z=415.3 [M+H].sup.+.

(37) Step 9: Synthesis of Compounds WX002A and WX002B

(38) Compound 2-9 (200 mg, 482.64 μmol, 1 eq) was dissolved in tetrahydrofuran (2 mL), then tri-n-butylphosphine (195.29 mg, 965.28 μmol, 238.16 μL, 2 eq) was added, the resulting reaction mixture was cooled to 0° C. 1,1′-(azodicarbonyl)dipiperidine (243.55 mg, 965.28 μmol, 2 eq) was added, and the resulting reaction mixture was allowed to react at 25° C. for 4 hours. The above reaction was conducted in duplicate, and the 2 batches of reaction mixture were combined and then directly concentrated to dryness. The residue was purified by flash silica gel column chromatography (petroleum ether/ethyl acetate=0 to 90%) and preparative plate (ethyl acetate:methanol=10:1) to obtain compound WX002. The compound WX002 was resolved by SFC (column: DAICEL CHIRALCEL OD-H (250 mm*30 mm, 5 μm); mobile phase: A (CO.sub.2) and B (methanol, containing 0.1% ammonium hydroxide); gradient: B %=32%-32%, 7.5 min, to obtain WX002A and WX002B.

(39) WX002A: .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.27 (s, 1H), 8.42 (d, J=7.6 Hz, 1H), 8.30 (s, 1H), 7.97 (d, J=2.8 Hz, 1H), 7.59-7.57 (m, 1H), 6.79 (d, J=8.0 Hz, 1H), 6.11 (t, J=8.4 Hz, 1H), 4.88 (d, J=10.8 Hz, 1H), 4.53 (t, J=8.0 Hz, 1H), 0.33.97-3.90 (m, 1H), 3.84 (d, J=10.8 Hz, 1H), 3.08-3.01 (m, 1H), 2.60-2.46 (m, 1H), 2.39-2.33 (m, 1H), 1.48-1.42 (m, 1H), 0.95-0.90 (m, 1H), 0.87-0.81 (m, 1H). LCMS m/z=397.3 [M+H].sup.+.

(40) SFC (column: Chiralcel OD-3, 3 μm, 0.46 cm id×10 cm L; mobile phase: A (CO.sub.2) and B (MeOH, containing 0.05% isopropylamine); gradient: B %=5 to 40%, 5 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt=2.14 min, 100% excess of chiral isomer.

(41) WX002B: .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.27 (s, 1H), 8.41 (d, J=7.6 Hz, 1H), 8.30 (s, 1H), 7.97 (d, J=2.8 Hz, 1H), 7.59-7.56 (m, 1H), 6.79 (d, J=7.6 Hz, 1H), 6.13-6.09 (m, 1H), 4.88 (dd, J=10.8, 1.6 Hz, 1H), 4.53 (t, J=8.0 Hz, 1H), 3.97-3.90 (m, 1H), 3.84 (d, J=10.8 Hz, 1H), 3.08-3.01 (m, 1H), 2.60-2.49 (m, 1H), 2.39-2.33 (m, 1H), 1.48-1.42 (m, 1H), 0.96-0.90 (m, 1H), 0.87-0.81 (m, 1H). LCMS m/z=397.3 [M+H].sup.+.

(42) SFC (column: Chiralcel OD-3, 3 μm, 0.46 cm id×10 cm L; mobile phase: A (CO.sub.2) and B (MeOH, containing 0.05% isopropylamine); gradient: B %=5 to 40%, 5 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt=2.49 min, 100% excess of chiral isomer.

Examples 5 to 6: Synthesis of Compounds WX003A and WX003B

(43) ##STR00038## ##STR00039##

(44) Step 1: Synthesis of Compound 3-2

(45) Compound 3-1 (20 g, 142.74 mmol, 1 eq) and imidazole (19.44 g, 285.49 mmol, 2 eq) were dissolved in dichloromethane (250 mL), and then a solution of tert-butyl dimethylchlorosilane (25.82 g, 171.29 mmol, 20.99 mL, 1.2 eq) in dichloromethane (30 mL) was slowly added dropwise at 0° C. After the dropwise addition, the reaction mixture was naturally warmed to 25° C. and the reaction was allowed to run for 15 hours. Additional imidazole (9.72 g, 142.74 mmol, 1 eq) and tert-butyl dimethylchlorosilane (10.76 g, 71.37 mmol, 8.75 mL, 0.5 eq) were added, and the reaction mixture was stirred at 25° C. for 12 hours. After completion of the reaction, the reaction mixture was poured into 300 mL of saturated aqueous solution of sodium bicarbonate, and then extracted with dichloromethane (300 mL each time). The organic phases were combined and washed with saturated brine (200 mL×3). The organic phase was dried over an appropriate amount of anhydrous sodium sulfate, filtered to remove the desiccant, and concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography to obtain compound 3-2. .sup.1H NMR: (400 MHz, CDCl.sub.3) δ: 10.39-10.37 (m, 1H), 7.46 (dd, J=3.2, 8.0 Hz, 1H), 7.20-7.11 (m, 1H), 6.87-6.85 (m, H), 1.01 (s, 9H), 0.26 (dd, J=2.4, 3.6 Hz, 6H).

(46) Step 2: Synthesis of Compound 3-3

(47) Compound 3-2 was dissolved in ethyl acetate (450 mL), and then 2,2-dimethyl-1,3-dioxane-4,6-dione (13.48 g, 93.56 mmol, 1 eq), tert-butyl N-hydroxycarbamate (12.46 g, 93.56 mmol, 1 eq) and 1,4-diazabicyclo[2.2.2]octane (1.05 g, 9.36 mmol, 1.03 mL, 0.1 eq) were added. The resulting reaction mixture was stirred at 25° C. for 18 hours under nitrogen atmosphere. After completion of the reaction, the reaction mixture was washed with water (50 mL) and saturated brine (50 mL×2). The organic phase was dried over an appropriate amount of anhydrous sodium sulfate. The desiccant was removed by filtration, and the filtrate was concentrated to dryness to obtain a crude yellow oily product. The crude product was purified by column chromatography to obtain compound 3-3.

(48) .sup.1H NMR: (400 MHz, CDCl.sub.3) δ: 7.16 (dd, J=3.2, 9.2 Hz, 1H), 6.92-6.87 (m, 1H), 6.77 (dd, J=4.8, 8.8 Hz, 1H), 5.75 (dd, J=3.2, 9.6 Hz, 1H), 3.31-3.25 (m, 1H), 2.71 (dd, J=3.2, 17.6 Hz, 1H), 1.51 (s, 9H), 1.01 (s, 9H), 0.27 (d, J=12.4 Hz, 6H). LCMS m/z=434 [M+23].sup.+, 311.9 [M−100+H].sup.+.

(49) Step 3: Synthesis of Compound 3-4

(50) Compound 3-3 (2.01 g, 4.88 mmol, 1 eq) was dissolved in tetrahydrofuran (20 mL), then lithium borohydride (319.18 mg, 14.65 mmol, 3 eq) was added, and the resulting reaction mixture was stirred at 12° C. for 0.5 hour. After completion of the reaction, 10 mL of saturated solution of ammonium chloride was slowly added to the reaction mixture to quench the reaction. The mixture was stirred for 20 minutes, and then extracted with ethyl acetate (50 mL×3). The organic phases were combined, dried over an appropriate amount of anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated to dryness to obtain compound 3-4. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 7.50 (br s, 1H), 7.42-7.32 (m, 1H), 6.87-6.77 (m, 1H), 6.76-6.63 (m, 1H), 5.53-5.42 (m, 1H), 3.90-3.71 (m, 2H), 3.25 (br s, 1H), 2.32-2.16 (m, 1H), 2.10-2.01 (m, 1H), 1.44-1.36 (m, 9H), 1.07-0.98 (m, 9H), 0.27 (d, J=1.2 Hz, 6H). LCMS m/z=438.1 [M+23].sup.+, 316 [M−100+H].sup.+.

(51) Step 4: Synthesis of Compound 3-5

(52) Compound 3-4 (23 g, 55.35 mmol, 1 eq) and triphenylphosphine (36.29 g, 138.36 mmol, 2.5 eq) were dissolved in anhydrous tetrahydrofuran (300 mL), and the resulting solution was cooled to 0-5° C., followed by dropwise addition of diisopropyl azodicarboxylate (33.57 g, 166.04 mmol, 32.28 mL, 3 eq). After the dropwise addition, the ice bath was removed, and the reaction was allowed to run at 25° C. for 4 hours. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated to dryness to obtain a yellow oily liquid. 120 mL of a mixed solvent (ethyl acetate/petroleum ether=1:8) was added thereto, and the obtained mixture was stirred evenly, allowed to stand, and then filtered, the filter cake was rinsed with 50 mL of a mixed solvent (ethyl acetate/petroleum ether=8:1). The filtrate was collected and concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography to obtain compound 3-5. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 7.17 (dd, J=3.2, 9.6 Hz, 1H), 6.87-6.75 (m, 1H), 6.74-6.66 (m, 1H), 5.43 (dd, J=4.4, 8.4 Hz, 1H), 4.08-4.02 (m, 1H), 3.92 (q, J=8.0 Hz, 1H), 2.83-2.68 (m, 1H), 2.19-2.08 (m, 1H), 1.47 (s, 9H), 1.03 (s, 9H), 0.26 (d, J=10.8 Hz, 6H). LCMS m/z=420.0 [M+23].sup.+, 297.9 [M−100+H].sup.+.

(53) Step 5: Synthesis of Compound 3-6

(54) Compound 3-5 (2.03 g, 5.11 mmol, 1 eq) was dissolved in ethyl acetate (20 mL), a solution of hydrogen chloride in ethyl acetate (4 M, 7.66 mL, 6 eq) was added, and the resulting reaction mixture was stirred at 14° C. for 5 hours. After completion of the reaction, the reaction mixture was concentrated to dryness to obtain a crude product. The crude product was fully dispersed in a mixed solution (10 mL) of ethyl acetate/petroleum ether (10:1), the solid was collected by filtration, and dried under vacuum at 40° C. to obtain compound 3-6. .sup.1H NMR: (400 MHz, DMSO-d.sub.6) δ: 7.32-7.26 (m, 1H), 7.18-7.05 (m, 1H), 7.00-6.88 (m, 1H), 5.03-4.93 (m, 1H), 4.36-4.26 (m, 1H), 4.15-4.05 (m, 1H), 2.84-2.71 (m, 1H), 2.42-2.29 (m, 1H), 1.00 (s, 9H), 0.27 (d, J=3.2 Hz, 6H). LCMS m/z=297.9 [M+H].sup.+.

(55) Step 6: Synthesis of Compound 3-7

(56) Compound 3-6 (1.43 g, 4.28 mmol, 1 eq) and ethyl 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate (1.06 g, 4.71 mmol, 1.1 eq) were added into dimethyl sulfoxide (15 mL), followed by addition of triethylamine (1.30 g, 12.85 mmol, 1.79 mL, 3 eq). The resulting reaction mixture was allowed to react at 75° C. for 18 hours under nitrogen atmosphere. After completion of the reaction, the reaction mixture was concentrated to dryness. The residue was dissolved in 200 mL of ethyl acetate, and then washed with water (30 mL×3) and saturated brine (30 mL). The organic phase was dried over an appropriate amount of anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated to dryness to obtain a crude product as yellow solid. To the crude product was added 10 mL of ethyl acetate and 10 mL of petroleum ether, and the obtained mixture was slurried, and filtered to obtain the solid. The solid was dried under vacuum at 40° C. to obtain compound 3-7. .sup.1H NMR: (400 MHz, CDCl.sub.3) δ: 8.55 (s, 1H), 8.39 (d, J=8.0 Hz, 1H), 8.33 (s, 1H), 7.10-7.01 (m, 2H), 6.95-6.91 (m, 1H), 6.85 (d, J=7.6 Hz, 1H), 5.97 (t, J=7.2 Hz, 1H), 4.58-4.38 (m, 3H), 4.04-3.93 (m, 1H), 2.97-2.71 (m, 2H), 1.41 (t, J=7.2 Hz, 3H). LCMS m/z=373.0 [M+H].sup.+.

(57) Step 7: Synthesis of Compound 3-8

(58) Compound 3-7 (300 mg, 805.69 μmol, 1 eq), (1-tert-butoxycarbonylamino)cyclopropyl methyl methanesulfonate (277.90 mg, 1.05 mmol, 1.3 eq) and cesium carbonate (525.02 mg, 1.61 mmol, 2 eq) were added into N,N-dimethylformamide (2 mL), and the resulting reaction mixture was stirred at 80° C. for 5 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with ethyl acetate (200 mL), and filtered through diatomite. The filtrate was washed with water (20 mL×3). The organic phase was dried over an appropriate amount of anhydrous sodium sulfate, and filtered to remove the desiccant, and the filtrate was concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography to obtain compound 3-8. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 8.45 (d, J=7.6 Hz, 1H), 8.34 (s, 1H), 7.21 (dd, J=2.9, 9.2 Hz, 1H), 7.02 (d, J=7.6 Hz, 1H), 6.94-6.87 (m, 1H), 6.84-6.78 (m, 1H), 6.30-6.22 (m, 1H), 5.91 (br s, 1H), 4.38-4.25 (m, 2H), 4.19-4.01 (m, 2H), 3.97-3.80 (m, 1H), 2.95-2.82 (m, 1H), 2.48-2.37 (m, 1H), 1.70-1.57 (m, 14H), 1.07-0.78 (m, 4H). LCMS m/z=542.3 [M+H].sup.+.

(59) Step 8: Synthesis of Compound 3-9

(60) Compound 3-8 (150 mg, 276.97 μmol, 1 eq) was dissolved in methanol (3 mL), then sodium hydroxide solution (2 M, 830.92 μL, 6 eq) was added, and the resulting reaction mixture was stirred at 60° C. for 18 hours. After completion of the reaction, the reaction mixture was concentrated to dryness, and the residue was added with water (5 mL) and stirred until completely dissolved. The pH value of the obtained solution was adjusted to 4-5 with 1 M hydrochloric acid, and then the mixture was extracted with ethyl acetate (20 mL×3). The organic phases were combined, dried over an appropriate amount of anhydrous sodium sulfate, and filtered to remove the desiccant. The filtrate was concentrated to dryness to obtain a crude product of compound 3-9. LCMS m/z=514.1 [M+H].sup.+.

(61) Step 9: Synthesis of Compound 3-10

(62) Compound 3-9 (143 mg, 278.47 μmol, 1 eq) was dissolved in ethyl acetate (3 mL), and then a solution of hydrogen chloride in ethyl acetate (4 M, 69.62 μL, 1 eq) was added. The resulting reaction mixture stirred at 13° C. for 18 hours. After completion of the reaction, the reaction mixture was concentrated to dryness to obtain product 3-10. LCMS m/z=414.1 [M+H].sup.+.

(63) Step 10: Synthesis of Compound 3-11

(64) Compound 3-10 (128 mg, 309.63 μmol, 1 eq) and N,N-diisopropylethylamine (200.09 mg, 1.55 mmol, 269.66 μL, 5 eq) were added into a mixed solvent of dichloromethane (20 mL) and N,N-dimethylformamide (4 mL), followed by addition of pentafluorophenyl diphenylphosphinate (154.66 mg, 402.51 μmol, 1.3 eq). The resulting reaction mixture was stirred at 25° C. for 4 hours. After completion of the reaction, 3M aqueous solution of sodium carbonate (3 mL) was added to the reaction mixture and stirred for 5 minutes, and then extracted and ethyl acetate (100 mL). The aqueous layer was discarded, the organic phase was washed with saturated brine (15 mL×3), then dried over an appropriate amount of anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated to dryness to obtain a brown oily liquid. The crude product was purified by column chromatography (ethyl acetate/petroleum ether=0 to 45%) to obtain compound 3-11. LCMS m/z=396.1 [M+H].sup.+.

(65) Step 11: Synthesis of Compounds WX003A and WX003B

(66) Compound 3-11 (180 mg, 455.25 μmol, 1 eq) was resolved by supercritical fluid chromatography (SFC) (column: Phenomenex-Amylose-1 (250 mm*30 mm, 5 μm); mobile phase: A (CO.sub.2) and B (ethanol, containing 0.1% ammonium hydroxide); gradient: B %=40%-40%, 10 min) to obtain compounds WX003A and WX003B.

(67) WX003A: .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.51 (s, 1H), 8.40 (d, J=7.2 Hz, 1H), 8.30 (s, 1H), 7.21 (dd, J=3.2, 9.2 Hz, 1H), 6.99-6.93 (m, 1H), 6.83-6.75 (m, 2H), 6.37-6.30 (m, 1H), 4.53 (t, J=7.6 Hz, 1H), 4.41 (dd, J=2.0, 9.2 Hz, 1H), 3.93-3.85 (m, 1H), 3.73 (d, J=9.2 Hz, 1H), 3.08-2.97 (m, 1H), 2.65-2.53 (m, 2H), 1.32-1.28 (m, 1H), 0.93-0.82 (m, 2H). LCMS m/z=396.2 [M+H].sup.+. SFC (Column: Chiralpak AD-3 150×4.6 mm I.D., 3 μm; Mobile phase: A (CO.sub.2) and B (ethanol, containing 0.05% diethylamine); gradient: B %=40%, 6 min; flow rate: 2.5 mL/min; column temperature: 35° C.), Rt=3.689 min, 100% isomer excess.

(68) WX003B: .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.51 (s, 1H), 8.40 (d, J=7.2 Hz, 1H), 8.30 (s, 1H), 7.21 (dd, J=3.2, 9.2 Hz, 1H), 6.99-6.93 (m, 1H), 6.83-6.75 (m, 2H), 6.37-6.30 (m, 1H), 4.53 (t, J=7.6 Hz, 1H), 4.41 (dd, J=2.0, 9.2 Hz, 1H), 3.93-3.85 (m, 1H), 3.73 (d, J=9.2 Hz, 1H), 3.08-2.97 (m, 1H), 2.65-2.53 (m, 2H), 1.32-1.28 (m, 1H), 0.93-0.82 (m, 2H). LCMS m/z=396.2 [M+H].sup.+. SFC (Column: Chiralpak AD-3 150×4.6 mm I.D., 3 μm; Mobile phase: A (CO.sub.2) and B (ethanol, containing 0.05% diethylamine); gradient: B %=40%, 6 min; flow rate: 2.5 mL/min; column temperature: 35° C.), Rt=4.561 min, 99.74% isomer excess.

(69) By methods similar to the synthesis methods of steps 1-9 in Example 2, the examples in the following table were synthesized. The SFC conditions of each example in the following table are the resolution conditions of chiral carbon in isoxazolyl, and the remaining chiral carbons in the structural formulas are directly introduced from the corresponding raw materials in the synthetic process.

(70) TABLE-US-00001 Ex- MS am- Com- m/z: ple pound Structure NMR and SFC [M + H].sup.+  7 WX004A 0 .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.66 (d, J = 7.6 Hz, 1 H), 8.25 (s, 1 H), 8.01 (d, J = 2.8 Hz, 1 H), 7.77-7.74 (m, 1H), 6.91 (d, J = 8.0 Hz, 1 H), 6.08-6.03 (m, 1 H), 4.66-4.55 (m, 2 H), 4.39-4.33 (m, 2 H), 3.98-3.92 (m, 1 H), 3.09-3.03 (m, 1 H), 2.66-2.56 (m, 1 H), 1.50 (d, J = 6.4 Hz, 3 H) 385.1  8 WX004B .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.31 (s, 1 H), 8.45 (s, 1H), 8.00 (d, J = 2.4 Hz, 1 H), 7.61-7.59 (m, 1 H), 6.86 (d, J = 7.6 Hz, 1 H), 6.06 (s, 1H), 4.71-4.66 (m, 1H), 4.52 (t, J = 7.6, 14.8 Hz, 1 H), 4.36-4.26 (m, 2H), 3.83 (s, 1H), 3.43 (s, 1H), 2.99 (s, 1H), 2.53 (s, 1H), 1.61 (s, 3H). 385.1  9 WX005A .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.20 (s, 1 H), 8.44 (d, J = 7.6 Hz, 1 H), 8.37 (s, 1 H), 7.95 (d, J = 3.2 Hz, 1 H), 7 54 (dd, J = 2.8, 8.0 Hz, 1 H), 6.81 (d, J = 12.4 Hz, 1 H), 5.98-5.94 (m, 1 H), 4.99- 4.92 (m, 1 H), 4.54-4.50 (m, 1 H), 4.05-3.96 (m, 2 H), 3.94- 3.78 (m, 1 H), 2.58-2.50 (m, 1H), 1.60 (d, J = 6.4 Hz, 3 H). SFC (column: Chiralpak AD-H, 5 μm, 3 cm id × 25 cm L; mobile 385.1 phase: A (CO.sub.2) and B (EtOH, containing 0.1% ammonium hydroxide); gradient: B% = 50%, flow rate: 70 g/min; wavelength: 220 nm; pressure: 100 bar., column temperature: 40° C., Rt = 4.3 min. 10 WX005B .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.65 (d, J = 8.0 Hz, 1 H), 8.23 (s, 1 H), 8.00 (d, J = 3.2 Hz, 1 H), 7.73 (dd, J = 3.2, 8.8 Hz, 1 H), 6.89 (d, J = 7.6 Hz, 1 H), 5.94- 5.90 (m, 1 H), 5.34-5.26 (m, 1 H), 4.58 (t, J = 6.8, 14.4 Hz, I H), 4.07-3.97 (m, 2 H), 3.40 (dd, J = 8.4, 13.6 Hz, 1 H), 3.07-3.01 (m, 1H), 2.62-2.52 (m, 1 H), 1.55 (d, J = 6.4 Hz, 3 H). SFC (column: Chiralpak AD-H, 385.1 5 μm, 3 cm id × 25 cm L; mobile phase: A (CO.sub.2) and B (EtOH, containing 0.1% ammonium hydroxide); gradient: B% = 50%, flow rate: 70 g/min; wavelength: 220 nm; pressure: 100 bar., column temperature: 40° C., Rt = 4.97 min. 11 WX006A .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.17 (s, 1H), 8.45-8.41 (m, 2 H), 8.00 (d, J = 2.4 Hz, 1 H), 7.60- 7.57 (m, 1 H), 6.82 (d, J = 7.6 Hz, 1 H), 5.97-5.93 (m, 2 H), 5.52 (d, J = 5.4 Hz, 1 H), 4.99 (d, J = 11.2 Hz, 1 H), 4.76 (d, J = 11.2 Hz, 1 H), 4.63 (d, J = 5.4 Hz, 2 H), 4.53 (t, J = 7.6 Hz, 1 H), 3.92-3.85 (m, 1 H), 3.04-2.98 (m, 1 H), 2.60- 2.52 (m, 1 H). SFC (column: Chiralcel OD-3, 3 μm, 0.46 cm id × 10 cm L; mobile 413.1 phase: A (CO.sub.2) and B (EtOH, containing 0.05% isopropylamine); gradient: B% = 5 to 40%, 5 min; flow rate: 4.0 mL/ min; wavelength: 220 nm; pressure: 100 bar, Rt = 2.7 min, chiral isomer excess 100%. 12 WX006B .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.17 (s, 1H), 8.45-8.41 (m, 2 H), 8.00 (d, J = 2.4 Hz, 1 H), 7.60- 7.57 (m, 1 H), 6.82 (d, J = 7.6 Hz, 1 H), 5.97-5.93 (m, 2 H), 5.52 (d, J = 5.4 Hz, 1 H), 4.99 (d, J = 11.2 Hz, 1 H), 4.76 (d, J = 11.2 Hz, 1 H), 4.63 (d, J = 5.4 Hz, 2 H), 4.53 (t, J = 7.6 Hz, 1 H), 3.92-3.85 (m, 1 H), 3.04-2.98 (m, 1 H), 2.60- 2.52 (m, 1 H). SFC (column: Chiralcel OD-3, 3 μm, 0.46 cm id × 10 cm L; mobile 413.1 phase: A (CO.sub.2) and B (EtOH, containing 0.05% isopropylamine); gradient: B% = 5 to 40%, 5 min; flow rate: 4.0 mL/ min; wavelength: 220 nm; pressure: 100 bar, Rt = 3.41 min, chiral isomer excess 100%. 13 WX007A .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.57 (d, J = 6.8 Hz, 1 H), 8.41 (d, J = 7.8 Hz, 1 H), 8.36 (s, 1H), 7.94 (d, J = 3.2 Hz, 1 H), 7.54 (dd, J = 8.4, 3.2 Hz, 1 H), 6.77 (d, J = 7.8 Hz, 1 H), 5.98-5.93 (m, 1 H), 5.64-5.60 (m, 1 H), 4.55- 4.51 (m, 1 H), 4.45-4.40 (m, 1 H), 3.99-3.94 (m, 1 H), 3.06- 3.01 (m, 1 H), 2.58-2.50 (m, 1 H), 2.37-2.23 (m, 2 H), 2.02- 1.81 (m, 3 H), 1.71-1.68 (m, 1 H). 411.1 SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (CO.sub.2) and B (MeOH, containing 0.05% isopropylamine); gradient: B% = 10 to 40%, 3 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.48 min, chiral isomer excess 100%. 14 WX007B .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.67 (d, J = 6.8 Hz, 1 H), 8.50 (d, J = 7.8 Hz, 1 H), 8.44 (s, 1H), 8.00 (d, J = 3.2 Hz, 1 H), 7.58 (dd, J = 8.4, 3.2 Hz, 1 H), 6.90 (d, J = 7.8 Hz, 1 H), 6.27-6.23 (m, 1 H), 5.19-5.14 (m, 1 H), 4.51 (t, J = 8.4 Hz, 1 H), 4.35 (s, 1 H), 3.75-3.69 (m, 1 H), 3.04-2.99 (m, 1 H), 2.95-2.88 (m, 1 H), 2.72-2.51 (m, 2 H), 1.90-1.62 (m, 3 H), 1.54-1.46 (m, 1 H). SFC (column: Chiralpak AS-3, 411.1 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (CO.sub.2) and B (MeOH, containing 0.05% isopropylamine); gradient: B% = 10 to 40%, 3 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.73 min, chiral isomer excess 100%. 15 WX008A .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.60 (d, J = 6.4 Hz, 1H), 8.43 (d, J = 7.6 Hz, 1H), 8.37 (s, 1H), 7.94 (d, J = 2.8 Hz, 1H), 7.57 (dd, J = 8.0, 2.8 Hz, 1H), 6.79 (d, J = 7.2 Hz, 1H), 5.99-5.94 (m, 2H), 4.77-4.70 (m, 2H), 4.34-4.24 (m, 2H), 4.11-4.07 (m, 1H), 4.03-3.96 (m, 1H), 3.89 (dd, J = 7.2, 1.6 Hz, 1H), 3.09-3.02 (m, 1H), 2.60-2.50 (m, 1H). SFC (column: Chiralcel OD-3, 3 μm, 0.46 cm id × 5 cm L; mobile 413.1 phase: A (CO.sub.2) and B (MeOH, containing 0.05% isopropylamine); gradient: B% = 10 to 40%, 3 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.67 min, chiral isomer excess 100%. 16 WX008B .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 10.01 (s, 1H), 8.52 (d, J = 7.6 Hz, 1H), 8.43 (s, 1H), 7.97 (d, J = 2.8 Hz, 1H), 7.62 (dd, J = 8.0, 2.8 Hz, 1H), 6.92 (d, J = 7.6 Hz, 1H), 6.25-6.21 (m, 1H), 5.34-5.39 (m, 1H), 4.75-4.67 (m, 2H), 4.54-4.50 (m, 2H), 4.02 (dd, J = 10.0, 3.6 Hz, 1H), 3.77-3.67 (m, 2H), 2.96-2.88 (m, 1H), 2.63- 2.53 (m, 1H) SFC (column: Chiralcel OD-3, 3 μm, 0.46 cm id × 5 cm L; mobile 413.1 phase: A (CO.sub.2) and B (MeOH, containing 0.05% isopropylamine); gradient: B% = 10 to 40%, 3 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.94 min, chiral isomer excess 98.72%. 17 WX009A 0 .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.68-8.66 (m, 1H), 8.24 (s, 1H), 8.03 (d, J = 2.8 Hz, 1H), 7.84- 7.82 (m, 1H), 6.94-6.89 (m, 1H), 6.25-6.23 (m, 1H), 6.06-6.02 (m, 1H), 5.07-5.03 (m, 1H), 4.63-4.60 (m , 1H), 4.17-3.92 (m, 3H), 3.73-3.65 (m, 2H), 3.12-3.05 (m, 4H), 2.69-2.59 (m, 1H). SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 10 cm L; mobile phase: A (CO.sub.2) and B (MeOH, containing 0.05% isopropylamine); gradient: B% = 426.1 10 to 40%, 5 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 2.36 min, chiral isomer excess 100%. 18 WX009B .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.76 (d, J = 7.6 Hz, 1H), 8.32 (s, 1H), 8.08 (d, J = 3.2 Hz, 1H), 7.94-7.91 (m, 1H), 7.01 (d, J = 7.6 Hz, 1H), 6.03 (t, J = 7.6 Hz, 1H), 5.69-5.64 (m, 1H), 4.92- 4.90 (m, 1H), 4.59-4.56 (m, 1H), 4.51-4.48 (m, 1H), 4.06-3.90 (m, 3H), 3.59-3.55 (m, 1H), 3.04 (s, 3H), 2.980-2.95 (m, 1H), 2.70-2.67 (m, 1H). SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 10 cm L; mobile phase: A (CO.sub.2) and B (MeOH, containing 0.05% 426.1 isopropylamine); gradient: B% = 10 to 40%, 5 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 2.72 min, chiral isomer excess 100%. 19 WX010A .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.64 (s, 1H), 8.52 (d, J = 7.6 Hz, 1H), 8.44 (s, 1H), 8.00 (d, J = 2.8 Hz, 1H), 7.60 (dd, J = 8.4, 2.8 Hz, 1H), 6.91 (d, J = 7.6 Hz, 1H), 6.31-6.28 (m, 1H), 5.58 (d, J = 12.0 Hz, 1H), 5.17-5.12 (m, 1H), 4.53-4.49 (m, 1H), 4.08 (s, 1H), 3.98-3 94 (m, 1H), 3.75- 3.69 (m, 1H), 3.58-3.53 (m, 2H), 2.94-2.87 (m, 1H), 2.62-2.54(m, 1H), 2.52-2.42 (m, 1H), 1.81- 1.77 (m, 1H). SFC (column: Chiralpak AD-3, 427.1 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (CO.sub.2) and B (EtOH, containing 0.05% isopropylamine); gradient: B% = 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.93 min, chiral isomer excess 100.00%. 20 WX010B .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.63 (d, J = 8.8 Hz, 1H), 8.43 (d, J = 7.6 Hz, 1H), 8.36 (s, 1H), 7.97 (d, J = 2.8 Hz 1H), 7.59-7.57 (m, 1H), 6.78 (d, J = 7.6 Hz, 1H), 5.97-5.93 (m, 1H), 5.34 (d, J = 2.8 Hz, 1H), 4.58-4.54 (m, 1H), 4.41-4.34 (m, 1H), 4.12 (dd, J = 10.8, 5.2 Hz, 1H), 4.00- 3.94 (m, 1H), 3.91-3.87 (m, 1H), 3.74-3.69 (m, 1H), 3.57 (t, J = 10.8 Hz, 1H), 3.11-3.05 (m, 1H), 2.61-2.44 (m, 2H), 2.18- 2.11 (m, 1H). 427.1 SFC (column: Chiralpak AD-3, 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (CO.sub.2) and B (EtOH, containing 0.05% isopropylamine); gradient: B% = 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 2.31 min, chiral isomer excess 96.53%. 21 WX011A .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.76 (d, J = 8.8 Hz, 1 H), 8.41 (d, J = 7.6 Hz, 1 H), 8.37 (s, 1 H), J = 7.95 (d, J = 3.2 Hz, 1 H), 7.57- 7.54 (m, 1 H), 8 = 6.76 (d, J = 7.6 Hz, 1 H), 5.97-5.93 (m, 1 H), 5.11 (s, 1 H), 4.52 (t, J = 6.8 Hz, 1 H), 4 16-4.13 (m, 1 H), 3.96- 3.92 (m, 1 H), 3.92-3.71 (m, 1 H), 3.19-3.14 (m, 1 H), 2.98- 2.95 (m, 1 H), 2.56-2.46 (m, 1 H), 2.43-2.37 (m, 1 H), 2.35 (s, 3 H), 2.24-2.04 (m, 3 H). SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 5 cm L; mobile 440.1 phase: A (CO.sub.2) and B (iPrOH, containing 0.05% isopropylamine); gradient: B% = 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.78 min, chiral isomer excess 100%. 22 WX011B .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.63 (s, J = 8.8 Hz, 1 H), 8.52 (d, J = 7.6 Hz, 1 H), 8.44 (s, 1 H), J = 8.01 (d, J = 2.8 Hz, 1 H), 7.57 (dd, J = 8.4, 3.2 Hz, 1 H), J = 6.91 (d, J = 7.6 Hz, 1 H), 6.31-6.27 (m, 1 H), 5.17-5.12 (m, 1 H), 4.51 (t, J = 6.8 Hz, 1 H), 4.27- 4.25 (m, 1 H), 3.75-3.68 (m, 1 H), 3.47-3.44 (m, 1 H), 2.91- 2.86 (m, 1 H), 2.70-2.67 (m, 1 H), 2.63-2.53 (m, 1 H), 2.29- 2.26 (m, 1 H), 2.23 (s, 3 H), 2.01- 1.96 (m, 1 H), 1.93-1.80 (m, 2 H). 440.1 SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (CO.sub.2) and B (iPrOH, containing 0.05% isopropylamine); gradient: B% = 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 2.07 min, chiral isomer excess 98.53%. 23 WX012A .sup.1HNMR (400 MHz, CDCl.sub.3) δ: 8.66-8.64 (m, 1H), 8.22 (s, 1H), 8.04-8.03 (m, 1H), 7.84-7.81 (m, 1H), 6.90-6.88 (m, 1H), 6.05-5.96 (m, 2H), 4.95 (s, 1H), 4.60 (t, J = 7.6 Hz, 1H), 4.05- 3.99 (m, 1H), 3.91-3.78 (m, 3H), 3.59-3.55 (m, 1H), 3.08 (s, 1H), 2.67-2.58 (m, 1H). SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (CO.sub.2) and B (MeOH, containing 0.05% isopropylamine); gradient: B% = 412.1 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.99 min, chiral isomer excess 100%. 24 WX012B .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.67-8.64 (m, 1H), 8.23-8.22 (m, 1H), 8.04 (d, J = 2.8 Hz, 1H), 7.83 (dd, J = 8.4, 2.8 Hz, 1H), 6.91-6.89 (m, 1H), 6.06-5.97 (m, 2H), 4.95-4.93 (m, 1H), 4.62-4.58 (m, 1H), 4.05-3.99 (m, 1H), 3.91-3.78 (m, 3H), 3.59-3.48 (m, 1H), 3.11-3.04 (m, 1H), 2.67-2.58 (m, 1H). SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (CO.sub.2) and B (MeOH, containing 0.05% 412.1 isopropylamine); gradient: B% = 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.62 min, chiral isomer excess 98.59%. 25 WX013A .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.68 (d, J = 7.6 Hz, 1 H), 8.24 (s, 1 H), 8.05 (d, J = 3.2 Hz, 1 H), 7.84-7.81 (m, 1 H), 6.91 (d, J = 8 Hz, 1 H), 6.02-5.98 (m, 1 H), 5.36-5.35 (m, 1 H), 4.62-4.58 (m, 1 H), 4.44-4.39 (m, 1 H), 4.21-4.17 (m, 1 H), 4.03-3.97 (m, 1 H), 3.63-3.50 (m, 2 H), 3.28-3.23 (m, 1 H), 3.14-3.08 (m, 1 H), 2.67-2.57 (m, 1 H), 2.33-2.18 (m, 2 H). SFC (column: Chiralcel OD-3, 3 μm, 0.46 cm id × 5 cm L; mobile 426.1 phase: A (CO.sub.2) and B (MeOH, containing 0.05% isopropylamine); gradient: B% = 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 2.06 min, chiral isomer excess 100%. 26 WX013B .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.72 (d, J = 7.6 Hz, 1 H), 8.29 (s, 1 H), 8.14 (d, J = 4.0 Hz, 1 H), 7.97-7.94 (m, 1 H), 6.99-6.97 (m, 1 H), 5.94-5.90 (m, 1 H), 5.24-5.20 (m, 1 H), 4.61-4.57 (m, 1 H), 4.50-4.45 (m, 1 H), 4.06-4.00 (m, 1 H), 3.92-3.83 (m, 2 H), 3.62-3.59 (m, 1 H), 3.30-3.28 (m, 1 H), 3.03-2.96 (m, 1 H), 2.61-2.55 (m, 1 H), 2.36-2.30 (m, 2 H). SFC (column: Chiralcel OD-3, 3 μm, 0.46 cm id × 5 cm L; mobile 426.1 phase: A (CO.sub.2) and B (MeOH, containing 0.05% isopropylamine); gradient: B% = 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.78 min, chiral isomer excess 96.27%. 27 WX014A 0 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.72 (d, J = 46.4 Hz, 1 H), 8.55- 8.52 (m, 1 H), 8.43-8.41 (m, 1 H), 8.09-8.02 (m, 1 H), 7.72-7.56 (m, 1 H), 6.92 (d, J = 8.0 Hz, 1 H), 6.277-6.260 (m, 1 H), 5.32- 5.28 (m, 1 H), 5.05-4.90 (m, 1 H), 4.55-4.44 (m, 2 H), 4.42- 4.36 (m, 1 H), 3.77-3.65 (m, 1 H), 3.36-3.08 (m, 1 H), 2.95- 2.79 (m, 1 H), 2.65-2.56 (m, 2 H), 2.18-2.30 (s, 3 H), 1.35-1.25 (m, 2 H). SFC (column: Chiralpak AD-3, 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (CO.sub.2) and B (iPrOH, containing 0.05% 468.1 isopropylamine); gradient: B% = 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.99 min, chiral isomer excess 100%. 28 WX014B .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.78-9.71 (m, 1 H), 8.45-8.41 (m, 1 H), 8.35 (s, 1 H), 7.98-7.96 (m, 1 H), 7.64-7.59 (m, 1 H), 6.78 (d, J = 7.6 Hz, 1 H), 5.88- 5.82 (m, 1 H), 5.14-5.4.99 (m, 1 H), 4.79-4.76 (m, 1 H), 4.56- 4.53 (m, 1 H), 4.38-4.31 (m, 1 H), 3.96-3.90 (m, 1 H), 3.49- 3.45 (m, 1 H), 3.12-2.99 (m, 1 H), 2.76-2.70 (m, 1 H), 2.55- 2.47 (m, 1 H), 2.22-2.02 (m, 2 H), 1.93 (s, 3 H). SFC (column: Chiralpak AD-3, 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (CO.sub.2) and B (iPrOH, containing 0.05% 468.1 isopropylamine); gradient: B% = 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 2.47 min, chiral isomer excess 97.49%. 29 WX015A .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.71-8.66 (m, 1H), 8.26-8.25 (m, 1H), 8.04-8.03 (m, 1H), 7.84-7.82 (m, 1H), 6.95-6.90 (m, 1H), 6.21-6.19 (m, 1H), 6.07- 6.03 (m, 1H), 5.03 (s, 1H), 4.64-4.60 (m, 1H), 4.17-3.90 (m, 3H), 3.73-3.66 (m, 2H), 3.47-3.36 (m, 2H), 3.13-2.98 (m, 1H), 2.69-2.59 (m, 1H), 1.41 (t, J = 7.2 Hz, 3H). SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 10 cm L; mobile phase: A (CO.sub.2) and B (MeOH, containing 0.05% isopropylamine); gradient: B% = 440.1 10 to 40%, 5 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 2.31 min, chiral isomer excess 100%. 30 WX015B .sup.1H NMR (400 MHz, CD.sub.3OD) δ: 8.79-8.77 (m, 1H), 8.34 (s, 1H), 8.09 (d, J = 2.8 Hz, 1H), 7.95- 7.92 (m, 1H), 7.03 (d, J = 7.6 Hz, 1H), 6.10-6.06 (m, 1H), 5.69- 5.64 (m, 1H), 4.81-4.80 (m, 1H), 4.61-4.55 (m, 2H), 4.13-1.05 (m, 1H), 3.92-3.80 (m, 2H), 3.55-3.50 (m, 1H), 3.42-3.35 (m, 2H), 2.99-2.92 (m, 1H), 2.73-2.63 (m, 1H), 1.35 (t, J = 7.2 Hz, 3H). SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 10 cm L; mobile phase: A (CO.sub.2) and B (MeOH, containing 0.05% 440.1 isopropylamine); gradient: B% = 10 to 40%, 5 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 2.72 min, chiral isomer excess 100%. 31 WX016A .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.76 (d, J = 8.8 Hz, 1 H), 8.41 (d, J = 7.6 Hz, 1 H), 8.37 (s, 1 H), J = 7.95 (d, J = 3.2 Hz, 1 H), 7.57- 7.51 (m, 1 H), J = 6.76 (d, J = 7.6 Hz, 1 H), 5.98-5.94 (m, 1 H), 5.15 (s, 1 H), 4.52 (t, J = 6.8 Hz, 1 H), 4.25-4.14 (m, 1 H), 3.96- 3.90 (m, 1 H), 3.87-3.83 (m, 1 H), 3.25-3.14 (m, 1 H), 3.07- 2.99 (m, 1 H), 2.68-2.41 (m, 3 H), 2.38-2.31 (m, 1 H), 2.28- 2.07 (s, 3 H), 1.16-1.08 (m, 3 H). SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 5 cm L; mobile phase: A (CO.sub.2) and B (iPrOH, containing 0.05% 454.1 isopropylamine); gradient: B% = 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.77 min, chiral isomer excess 100%. 32 WX016B .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 9.63 (s, 1 H), 8.52 (d, J = 7.6 Hz, 1 H), 8.44 (s, 1 H), δ = 8.01 (d, J = 3.2 Hz, 1 H), 7.57 (dd, J = 8.4, 3.2 Hz, 1 H), δ = 6.92 (d, J = 7.6 Hz, 1 H), 6.32-6.28 (m, 1 H), 5.18-5.14 (m, 1 H), 4.52 (t, J = 6.8 Hz, 1 H), 4.29-4.27 (m, 1 H), 3.75-3.69 (m, 1 H), 3.57-3.44 (m, 2 H), 2.91-2.78 (m, 2 H), 2.63-2.53 (m, 1 H), 2.41-2.35 (m, 2 H), 2.24-2.18 (m, 1 H), 2.00 (t, J = 10.4 Hz, 1 H), 1.92- 1.78 (m, 1H), 1.02 (t, J = 7.2 Hz, 3 H). SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 5 cm L; mobile 454.1 phase: A (CO.sub.2) and B (iPrOH, containing 0.05% isopropylamine); gradient: B% = 5 to 40%, 4 min; flow rate: 4.0 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 2.05 min, chiral isomer excess 97.62%. 33 WX018 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 8.48 (d, J = 7.6 Hz, 1 H), 8.35 (s, 1 H), 7.53-7.50 (m, 1 H), 7.16 (s, I H), 7.02 (d, J = 7.6 Hz, 1 H), 5.95-5.92 (m, 1 H), 4.21-4.16 (m, 1 H), 4.07 (s, 2 H), 3.96-3.90 (m, 1 H), 2.81-2.77 (m, 1 H), 2.53-2.49 (m, 1 H), 1.49 (s, 3 H), 1.37 (s, 3 H). SFC (column: Chiralpak AS-3, 3 μm, 0.46 cm id × 10 cm L; mobile phase: A (CO.sub.2) and B (MeOH, 399.2 containing 0.05% isopropylamine); gradient: B% = 10 to 40%, 5 min; flow rate: 4 mL/min; wavelength: 220 nm; pressure: 100 bar, Rt = 1.83 min, chiral isomer excess 98.2%.

(71) By methods similar to the synthesis methods of steps 1 to 11 in Example 3, examples in the following table were synthesized.

(72) TABLE-US-00002 Ex- MS am- Com- m/z: ple pound Structure NMR and SFC [M + H].sup.+ 34 WX017A .sup.1H NMR (400 MHz, DMSO- d.sub.6) δ: 9.89 (d, J = 8.0 Hz, 1H), 8.95 (d, J = 7.2 Hz, 1H), 8.26 (br s, 1H), 7.33-6.99 (m, 3H), 6.85 (d, J = 7.2 Hz, 1H), 6.20- 6.10 (m, 1H), 4.60-4.50 (m, 1H), 4.47-4.40 (m, 1H), 4.29- 4.20 (m 1H), 4.09-3.92 (m, 2H), 3.91-3.82 (m, 1H), 3.03- 2.94 (m, 1H), 1.38 (d, J = 6.0 Hz, 3H). 384.1 35 WX017B .sup.1H NMR (400 MHz, DMSO- d.sub.6) δ: 9.33 (s, 1H), 8.97 (d, J = 7.6 Hz, 1H), 8.26 (s, 1H), 7.27-7.23 (m, 1H), 7.18-7.15 (m, 2H), 6.89 (d, J = 7.6 Hz, 1H), 6.15-6.08 (m, 1H), 4 55- 4.39 (m, 2H), 4.10-4.03 (m, 1H), 3.89-3.70 (m, 2H), 2.96- 2.87 (m, 1H), 2.44-2.36 (m, 1H), 1.48 (d, J = 6.3 Hz, 3H). 384.1

Experimental Example 1: Inhibitory Activities of the Compounds on TrkA, TrkC, ALK, Ros1 and Other Kinases

(73) The inhibitory activities of the compounds on TrkA, TrkC, ALK, Ros1 and other kinases were determined by Reaction Biology Corp. To reaction buffer (20 mM Hepes (pH 7.5), 10 mM MgCl.sub.2, 1 mM EGTA, 0.02% Brij35, 0.02 mg/mL BSA, 0.1 mM Na.sub.3VO.sub.4, 2 mM DTT, 1% DMSO) were sequentially added a certain concentration of substrate, coenzyme factor, kinase and test compound (10 concentrations, 3-fold serial dilutions, DMSO at a final concentration of 2%), and the mixture was mixed evenly. The mixture was incubated at room temperature for 20 minutes. To the reaction mixture was added a certain concentration of .sup.33P-ATP to initiate the reaction, and then the mixture was incubated at room temperature for 120 minutes. The radioactivities of the reactants were determined by the method of filter binding. The final kinase activity was expressed as the ratio of the remaining kinase activity in the test sample to the kinase activity in the DMSO control group. The dose-effect curve was fitted by GraphPad software and the IC.sub.50 was calculated. The results are shown in Table 1:

(74) TABLE-US-00003 TABLE 1 The half maximal inhibitory concentration IC.sub.50 on the kinases (nM) TrkA- TrkA- Ros- Compound TrkA G595R G667C TrkC ALK Ros1 G2032R WX001 177.00 / / / / / / hydrochloride WX002 4.37 / / / / / / WX002A 2.07 3.60 7.00 0.14 13.00 0.14 0.4 WX002B >1000 / / / / / / WX003A 2.66 3.59 3.04 0.06 28.10 0.15 / WX003B >1000 / / / / / / WX004A 14.30 38.50 42.00 0.41 109.00 0.94 / WX004B >1000 / / 105.00 / / / WX005A >1000 / / 189.00 >1000 531.00 / WX005B 13.10 10.70 26.40 0.09 19.40 0.35 2.65 WX006A 9.84 4.74 37.50 0.16 79.20 0.59 / WX006B >1000 >1000 / / / / / WX007A 2.80 5.18 2.85 0.05 11.60 0.13 0.73 WX007B 585.00 >1000 / / / / / WX008A 1.61 2.31 6.49 0.05 10.40 0.16 0.52 WX008B >1000 >1000 / / / / / WX009A 85.10 / / / / / / WX009B >1000 / / / / / / WX010A >1000 / / / / / / WX010B 3.10 / / / / / 0.61 WX011A 33.60 / / / / / / WX011B 270.00 / / / / / / WX012A >1000 / / / / / / WX012B 143.00 / / / / / / WX013A >1000 / / / / / / WX013B 57.20 / / / / / / WX014A 1230.00 / / / / / / WX014B 12.80 / / / / / / WX015A 182.00 / / / / / / WX015B >1000 / / / / / / WX016A 33.00 / / / / / / WX016B 220.00 / / / / / / WX017A 13.40 37.10 26.30 0.30 119.00 0.82 / WX017B >1000 / / / / / / WX018 >1000 / / / / / / LOXO-101 19.70 >1000 512.00 1.12 >1000 95.50 / LOXO-195 6.67 6.19 110.00 0.50 274.00 1.15 2.88 “/”: Not detected.

(75) The results show that the compounds of the present disclosure exhibit strong kinase inhibitory activities against a variety of kinases and their mutants, and exhibit strong inhibitory effects against gatekeeper mutation, solvent front mutation and DFG mutations of a variety of kinases.

Experimental Example 2: Inhibitory Activities of the Compounds on Cell Proliferation

(76) Adenosine Tri-Phosphate (ATP) is an energy carrier shared by various life activities in nature, and is the smallest unit of energy storage and transfer. The CellTiter-Glo™ cell viability detection kit uses luciferase as the detection substance, and luciferase requires the participation of ATP during the process of luminescence. CellTiter-Glo™ reagent is added to cell culture medium, and the luminescent intensity is measured. The luminescent signal is directly proportional to the amount of ATP in the system, and ATP is positively related to the number of living cells. Therefore, by using the CellTiter-Glo kit to detect ATP content, cell proliferation can be detected. In this assay, the cell line is Ba/F3 LMNA-NTRK1-WT stably transfected cell line, with 5000 cells/well.

(77) IC.sub.50 Determination Process:

(78) 1. Cell Culture and Inoculation

(79) a) the cells in the logarithmic growth phase were harvested, and counted with a platelet counter. Trypan blue exclusion method was used to detect cell viability to ensure the cell viability was no less than 90%.

(80) b) the cell concentration was adjusted; 90 μL of cell suspension was added respectively into a 96-well plate.

(81) c) the cells in the 96-well plates was cultured under conditions of 37° C., 5% CO.sub.2 and 95% humidity overnight.

(82) 2. Drug Dilution and Administration

(83) a) 10-fold drug solutions were prepared, the highest concentration was 10 μM, 9 concentrations, 3-fold diluted (refer to Appendix I). 10 μL of each drug solution was added to each well of the 96-well plate seeded with the cells in triplicate.

(84) b) the cells in the 96-well plate added with the drug were cultured under conditions of 37° C., 5% CO.sub.2 and 95% humidity for 72 hours, and then CTG (cell proliferation) was performed.

(85) 3. Reading at End Point

(86) a) CellTiter-Glo™ reagent was thawed, and the cell plate was equilibrated at room temperature for 30 minutes.

(87) b) CellTiter-Glo™ reagent of a same volume was added to each well;

(88) c) the cell plate was shaken on an orbital shaker for 5 minutes to lyse the cells.

(89) d) the cell plate was placed at room temperature for 20 minutes to stabilize the luminescent signal.

(90) e) the luminescent intensity was read.

(91) 4. Data Processing

(92) GraphPad Prism 5.0 software was used to analyze the data, nonlinear S-curve regression was used to fit the data to obtain dose-effect curve, and IC.sub.50 value was calculated therefrom. The data are shown in Table 2.

(93) TABLE-US-00004 TABLE 2 The half maximal inhibitory concentration IC.sub.50 against cells (nM) Ba/F3 Ba/F3 Ba/F3 BaF3 Ba/F3 LMNA- LMNA- LMNA- ETV6- Ba/F3 SLC34A2- NTRK1- NTRK1- NTRK1- NTRK3- SLC34A2- ROS1- Compound WT F589L G595R G623R ROS1-WT G2032R WX002A 1.26 0.78 6.20 3.40  2.53 16.08 WX003A 4.08 / 27.90 0.90 / / WX004A 7.69 / 14.50 2.10 / / WX005B 5.73 / 9.10 5.60 17.92 258.60  WX006A 23.50 / 19.40 5.40 / / WX007A 5.80 / 12.40 69.13  10.10 165.17  WX008A 1.60 1.59 3.92 3.49  5.45 38.47 WX009A 76.80 / / / / / WX010B 5.80 4.26 20.94 19.48  10.45 58.05 WX011A 39.71 / / / / / WX012B 258.75 / / / / / WX013B 215.55 / / / / / WX014B 80.41 / / / / / WX015A 65.13 / / / / / WX016A 65.80 / / / / / “/”: Not detected.

(94) The results show that the compounds of the present disclosure exhibit strong inhibitory activities on cell proliferation of Ba/F3 LMNA-NTRK1-WT stably transfected cell line. The compounds also exhibit strong inhibitory activities on cell proliferation of Ba/F3 LMNA-NTRK1-F589L, Ba/F3 LMNA-NTRK1-G595R, BaF3 ETV6-NTRK3-G623R, Ba/F3 SLC34A2-ROS1-WT and Ba/F3 SLC34A2-ROS1-G2032R stably transfected cell lines.

Experimental Example 3: In Vivo Cassette Pharmacokinetic Test of the Compounds in Mice

(95) Experimental object: male CD-1 mice aged 7-9 weeks were used as the experimental animals, the LC/MS/MS method was used to determine the drug concentrations of WX002A, TPX0005, Entrectinib (RXDX-101) and Larotrectinib (LOXO-101) in plasma and specific tissues at different time points after single intravenous (IV) and intragastric (PO) cassette administration of WX002A, TPX0005, Entrectinib (RXDX-101) and Larotrectinib (LOXO-101), in vivo pharmacokinetic behaviors of the compounds of the present disclosure in mice were studied, and pharmacokinetic characteristics thereof were evaluated.

(96) Drug preparation: WX002A, TPX0005, Entrectinib (RXDX-101) and Larotrectinib (LOXO-101) were prepared into clear solutions with 5% DMSO+10% solutol+85% water as solvent for administration in IV (intravenous) group and PO (intragastric) group. The dose of the compounds: IV dose of 1 mg/kg, administration volume of 2 mL/kg; PO dose of 3 mg/kg, and administration volume of 3 mL/kg. The results of pharmacokinetic parameters are shown in Table 3:

(97) TABLE-US-00005 TABLE 3 Results of in vivo cassette pharmacokinetic test in mice LOXO- RXDX- TPX- Compound 101 101 0005 WX002A IV Initial concentration 2379 2808 2253 3708 @ 1 C.sub.0 (nM) mpk Half life 1.21 1.94 3.69 0.88 T.sub.1/2 (h) Apparent volume 1.57 1.24 2.63 0.57 of distribution Vd (L/kg) Apparent clearance 22.7 7.9 16.5 6.3 Cl (mL/Kg/min) Area under curve 1717 3767 2857 6689 AUC.sub.0-inf (nM .Math. hr) PO Peak concentration 1353 586 1540 4740 @ 3 C.sub.max (nM) mpk Time of Peak 0.5 0.75 0.75 0.25 concentration T.sub.max (h) Area under curve 2720 3256 5494 19769 AUC.sub.0-inf (nM .Math. hr) Bioavailability 51% 23% 64% 76% F % drug concentration 40 ND 50 663 in brain at 0.5 h Brain@0.5 h (nmol/kg) drug concentration 20 30 41 343 in brain at 2 h Brain@2 h (nmol/kg) drug concentration 15 ND 8 63 in cerebrospinal fluid at 0.5 hour CSF@0.5 h (nmol/kg) drug concentration 3 ND ND 21 in cerebrospinal fluid at 2 hour CSF@2 h (nmol/kg) “ND”: Not detected.

(98) The results show that: WX002A has better pharmacokinetic properties in mice. Compared with TPX0005, Entrectinib (RXDX-101) and Larotrectinib (LOXO-101), the total exposure of WX002A after oral administration, and the exposure of WX002A in brain and cerebrospinal fluid CSF at 0.5 h and 2 h respectively after administration were significantly higher than the corresponding exposure of TPX0005, Entrectinib (RXDX-101) and Larotrectinib (LOXO-101) at the same dosage.

Experimental Example 4: The In Vivo Pharmacokinetic Test of the Compounds in Mice

(99) Experimental object: male CD-1 mice aged 7-9 weeks were used as the experimental animals, the LC/MS/MS method was used to determine the drug concentrations in plasma at different time points after single intravenous (IV) and intragastric (PO) administration of the compounds, in vivo pharmacokinetic behaviors of the compounds of the present disclosure in mice were studied, and pharmacokinetic characteristics thereof were evaluated.

(100) Drug preparation: The compounds were prepared into clear solutions with 5% DMSO+10% solutol+85% water as solvent for administration in IV (intravenous) group and PO (intragastric). The dose of the compounds: IV dose of 3 mg/kg, and PO dose of 10 mL/kg.

(101) The results of pharmacokinetic parameters are shown in Table 4:

(102) TABLE-US-00006 TABLE 4 Results of pharmacokinetic test in mice LOXO- LOXO- Compound 101 195 WX002A WX003A WX004A WX005B Half life IV 0.39 1.19 0.75 0.70 0.74 0.50 T.sub.1/2 (h) Apparent volume 0.97 0.50 0.49 0.81 0.75 0.44 of distribution Vd (L/kg) Apparent 37.4 11.8 7.2 10.8 10.4 12.4 clearance Cl (mL/Kg/min) Area under 3122 11146 17502 12216 12464 10544 the curve AUC.sub.0-last (nM .Math. hr) Peak PO 1548 12700 9145 7360 10520 17250 concentration C.sub.max (nM) Time of peak 0.38 0.38 0.50 0.38 0.5 0.38 concentration T.sub.max (h) Area under 2890 27344 29787 27967 29648 29367 the curve AUC.sub.0-last(nM .Math. hr) Bioavailability 28% 74% 51% 69% 72% 85% F % Compound WX006A WX007A WX008A WX010B WX017A Half life IV 0.56 0.88 1.14 0.92 0.71 T.sub.1/2 (h) Apparent volume 1.01 0.47 0.62 0.68 0.74 of distribution Vd (L/kg) Apparent 19.9 5.50 6.7 8.4 17.8 clearance Cl (mL/Kg/min) Area under 6104 22095 18220 14031 7337 the curve AUC.sub.0-last (nM .Math. hr) Peak PO 8775 22550 12200 13000 10790 concentration C.sub.max (nM) Time of peak 0.25 0.38 0.25 0.5 0.63 concentration T.sub.max (h) Area under 14448 54927 39553 49196 26657 the curve AUC.sub.0-last(nM .Math. hr) Bioavailability 72% 76% 65% 107% 111% F %

(103) The results show that the total exposure, peak concentration, and bioavailability of multiple compounds of the present disclosure after oral administration are significantly better than Larotrectinib (LOXO-101) and LOXO-195 at the same dosage, indicating excellent pharmacokinetics characteristics.

Experimental Example 5: Test of In Vivo Efficacy of the Compounds in Mice

(104) Experimental object: evaluation of the in vivo efficacy of test drugs such as WX002A on subcutaneous xenograft tumor of human colon cancer cell line KM12 cell in BALB/c mouse model.

(105) Drug preparation: The compounds were all prepared into clear solutions with 5% DMSO+10% solutol+85% water as solvent for administration in the PO (intragastric) group.

(106) Tumor measurement: the tumor diameters were measured with a vernier caliper twice a week. The calculation formula of tumor volume is: V=0.5×a×b.sup.2, wherein a and b represent the long diameter and short diameter of the tumor, respectively. The anti-tumor efficacy of the compounds is evaluated by TGI (%). TGI (%) reflects tumor growth inhibition rate. TGI (%)=[(1−(average tumor volume at the end of the administration in a treatment group−average tumor volume at the beginning of the administration in the treatment group))/(average tumor volume at the end of the treatment in the solvent control group−average tumor volume at the beginning of the treatment in the solvent control group)]×100%. The results are shown in FIG. 1.

(107) Statistical analysis: The statistical analysis was based on the relative tumor volume and tumor weight at the end of the experiment using SPSS software. One-way ANOVA was used to analyze the comparison between multiple groups. If the variance was uniform (the F values were not significantly different), the Tukey's method was used for analysis, and if the variance was not uniform (the F values were significantly different), the Games-Howell method was used for analysis. P<0.05 was considered to indicate significant difference.

(108) Experimental results: In the nude mouse xenograft model of human colon cancer KM12, the test compound WX002A had a significant anti-tumor effect at a dose as low as 3 mg/kg, and the anti-tumor effect had dose-effect-dependent trend (p<0.05 between the high-dose group and the low-dose group). The anti-tumor effect of WX002A at a dose of 3 mg/kg (T/C=33.18%, TGI=71.23%) and the anti-tumor effect of the compound LOXO-101 in the high-dose group (60 mg/kg) (T/C=34.20%, TGI=69.73%) were equivalent (P>0.05). The anti-tumor effect of WX002A at a dose of 15 mg/kg (T/C=15.63%, TGI=88.61%) was better than that of LOXO-101 high-dose group (60 mg/kg) (T/C=34.20%, TGI=69.73%), and was equivalent to the anti-tumor effect of TPX-0005 in the high-dose group (3 mg/kg) (T/C=16.80%, TGI=87.46%) (P>0.05).

Experimental Example 6: Test of the Efficacy of the Compounds in Mice

(109) Experimental object: evaluation of the in vivo efficacy of test drugs such as WX002A on subcutaneous xenograft tumor of human lung cancer LU-01-0414 in BALB/c mouse model.

(110) Drug preparation: The compounds were all prepared into clear solutions with 5% DMSO+10% solutol+85% water as solvent for administration in the PO (intragastric) group.

(111) Tumor measurement: the tumor diameters were measured with a vernier caliper twice a week. The calculation formula of tumor volume is: V=0.5×a×b.sup.2, wherein a and b represent the long diameter and short diameter of the tumor, respectively. The anti-tumor efficacy of the compounds was evaluated by TGI (%). TGI (%) reflects tumor growth inhibition rate. TGI (%)=[(1−(average tumor volume at the end of the administration in a treatment group−average tumor volume at the beginning of the administration in the treatment group))/(average tumor volume at the end of the treatment in the solvent control group−average tumor volume at the beginning of the treatment in the solvent control group)]×100%. The results are shown in FIG. 2.

(112) Statistical analysis: the statistical analysis was based on the relative tumor volume and tumor weight at the end of the experiment using SPSS software. One-way ANOVA was used to analyze the comparison between multiple groups. If the variance was uniform (the F values were not significantly different), the Tukey's method was used for analysis, and if the variance was not uniform (the F values were significantly different), the Games-Howell method was used for analysis. P<0.05 was considered to indicate significant difference.

(113) Experimental results: at the 14.sup.th day after administration in subcutaneous xenograft tumor of human lung cancer LU-01-0414, WX002A exhibits a significant inhibitory effect on tumor growth when administered at a dosage of 3, 15 and 30 mg/kg BID, with T/C of 9.57%, 3.07%, and 1.87% respectively, and TGI of 118.02%, 126.88%, and 128.36% respectively, and all WX002A groups show P<0.0001 compared with the solvent control group. Crizotinib administered at a dosage of 30 and 50 mg/kg QD exhibits T/C of 10.32% and 4.89% respectively, and TGI of 117.67% and 124.09% respectively, and all crizotinib groups show P<0.0001 compared with the solvent control group, indicating significant anti-tumor effect. The above results suggest that in the xenograft tumor model of human lung cancer LU-01-0414 in nude mice, WX002A has a significant anti-tumor effect at a dose as low as 3 mg/kg, and the anti-tumor effects of WX002A at a dose of 3 mg/kg and crizotinib at a dose of 30 mg/kg were equivalent (p>0.05).