SMAC MIMETICS USED AS IAP INHIBITORS AND USE THEREOF

Abstract

Disclosed are a class of SMAC mimetics used as IAP inhibitors, and in particular disclosed are compounds as shown in formula (I), isomers thereof, and pharmaceutically acceptable salts thereof. The IAP inhibitors are drugs for treating cancers, in particular breast cancer.

##STR00001##

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, ##STR00091## wherein, X.sub.1 is C(R.sub.5) or N; X.sub.2 is C(R.sub.6), N, O, or S; are each independently a single bond or a double bond; L is a single bond or —O—; R.sub.1 is —C(═O)NH.sub.2, CN, C.sub.1-5 alkyl, C.sub.1-5 heteroalkyl, phenyl, 5- to 6-membered heteroaryl, or 5- to 6-membered heterocycloalkyl; said C.sub.1-5 alkyl, C.sub.1-5 heteroalkyl, phenyl, 5- to 6-membered heteroaryl or 5- to 6-membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R; R.sub.2 is H, halogen, CN, COOH, —C(═O)NH.sub.2, C.sub.1-4 alkyl, or C.sub.1-4 heteroalkyl; said C.sub.1-4 alkyl or C.sub.1-4 heteroalkyl is optionally substituted with 1, 2 or 3 R; R.sub.3 and R.sub.7 are each independently H, halogen or C.sub.1-4 alkyl; said C.sub.1-4 alkyl is optionally substituted with 1, 2 or 3 R; R.sub.4 is H, phenyl, or 5- to 6-membered heteroaryl; R.sub.5 is H or halogen; R.sub.6 is H, halogen, C.sub.1-4 alkyl, C.sub.1-4 heteroalkyl, CN, or COOH; said C.sub.1-4 alkyl or C.sub.1-4 heteroalkyl is optionally substituted with 1, 2 or 3 R; R is halogen, OH, CN, CH.sub.3, CH.sub.3CH.sub.2, CH.sub.3CH.sub.2CH.sub.2, CH(CH.sub.3).sub.2, OCH.sub.3, OCF.sub.3, CHF.sub.2, CH.sub.2F, or NH.sub.2; and said C.sub.1-4 heteroalkyl, C.sub.1-5 heteroalkyl, 5- to 6-membered heterocycloalkyl and 5- to 6-membered heteroaryl each contain 1, 2 or 3 heteroatoms or heteroatom radicals independently selected from the group consisting of —NH—, —O—, —S—, N, —C(═O)O—, —C(═O)—, —C(═O)NH—, —C(═S)—, —S(═O)—, —S(═O)2—, —C(═NH)—, —S(═O).sub.2NH—, —S(═O)NH—, and —NHC(═O)NH—.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is a compound of formula (I′): ##STR00092## wherein X.sub.1, X.sub.2, L, R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.7 are as defined in claim 1.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein X.sub.2 is C(R.sub.6) or N.

4. (canceled)

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R.sub.1 is —C(═O)NH.sub.2, CN, CH.sub.3, CH.sub.3CH.sub.2, C.sub.1-5 alkyl-C(═O)—, C.sub.1-4 alkyl-C(═O)—, C.sub.1-5 alkyl-S(═O).sub.2—, C.sub.1-5 alkyl-N(H)C(═O)—, C.sub.1-4 alkyl-N(H)C(═O)—, (C.sub.1-2 alkyl)2—N—C(═O)—, phenyl, ##STR00093## said CH.sub.3, CH.sub.3CH.sub.2, C.sub.1-5 alkyl-C(═O)—, C.sub.1-4 alkyl-C(═O)—, C.sub.1-5 alkyl-S(═O).sub.2—, C.sub.1-5 alkyl-N(H)C(═O)—, C.sub.1-4 alkyl-N(H)C(═O)—, (C.sub.1-2 alkyl).sub.2-N—C(═O)—, phenyl, ##STR00094## is optionally substituted with 1, 2 or 3 R.

6. The compound according to claim 5 or a pharmaceutically acceptable salt thereof, wherein R.sub.1 is ##STR00095##

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R.sub.2 is H, halogen, C.sub.1-4 alkyl, or C.sub.1-4 alkyl-O—; said C.sub.1-4 alkyl or C.sub.1-4 alkyl-O— is optionally substituted with 1, 2 or 3 halogens.

8. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein R.sub.2 is H, F, Cl, Br, CF.sub.3, or OCF.sub.3.

9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R.sub.3 and R.sub.7 are each independently H, F, or Cl.

10. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R.sub.4 is H or ##STR00096##

11. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R.sub.5 is H or Cl.

12. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R.sub.6 is H, Cl, or CH.sub.3.

13. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein a structural unit ##STR00097##

14. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein a structural unit ##STR00098## is ##STR00099## ##STR00100## ##STR00101##

15. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is a compound of formula (II) or (III): ##STR00102## wherein R.sub.2, R.sub.3 and R.sub.7 are as defined in claim 1.

16. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, which is a compound of formula (II′) or (III′): ##STR00103## wherein R.sub.2, R.sub.3 and R.sub.7 are as defined in claim 2.

17. A compound or a pharmaceutically acceptable salt thereof, which is ##STR00104## ##STR00105## ##STR00106## ##STR00107## ##STR00108## ##STR00109## ##STR00110##

18. The compound according to claim 17 or a pharmaceutically acceptable salt thereof, which is ##STR00111## ##STR00112## ##STR00113## ##STR00114## ##STR00115## ##STR00116## ##STR00117##

19. A pharmaceutical composition, comprising a therapeutically effective amount of the compound according to claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient, and a pharmaceutically acceptable carrier.

20. A method for treating cancer, comprising administering an effective amount of the compound according to claim 1 or a pharmaceutically acceptable salt thereof to a subject in need thereof.

21. (canceled)

22. The method according to claim 20, wherein the cancer is breast cancer.

Description

DETAILED DESCRIPTION

[0119] The present disclosure is described in detail below by Examples, but it does not imply any disadvantageous limitation on the present disclosure. The present disclosure has been described in detail herein, and its specific embodiments are also disclosed therein. Various is changes and improvements made to the specific embodiments of the present disclosure without departing from the spirit and scope of the present disclosure will be apparent to a person skilled in the art.

##STR00044##

[0120] In the reactions shown in Reaction Scheme 1, Compound (A) is deprotected under an acidic condition (such as a hydrogen chloride/ethyl acetate solution) to obtain Compound (B); Compound (B) and Compound (C) undergo an acid-amine condensation reaction to obtain Compound (D), and this reaction requires a suitable condensing agent (such as HOBt), a suitable dehydrating agent (such as EDCI) and a suitable base (such as DIPEA) according to Reaction Scheme 1; and Compound (D) is then deprotected under an acidic condition (such as a hydrogen chloride/ethyl acetate solution) to obtain the compound represented by formula (I).

##STR00045##

[0121] In the reactions shown in Reaction Scheme 2, Compound (G) may be prepared via a substitution reaction between Compound (E) and Compound (F), this reaction requires a suitable base (such as potassium carbonate) and is preferably carried out at a high temperature according to Reaction Scheme 2; Compound (G) is deprotected under an acidic condition (such as a hydrogen chloride/ethyl acetate solution) to obtain Compound (H); and Compound (A) may be prepared via an acid-amine condensation reaction between Compound (H) and Compound (I), and this reaction requires a suitable condensing agent (such as HOBt), a suitable dehydrating agent (such as EDCI) and a suitable base (such as DIPEA).

##STR00046##

[0122] In the reactions shown in Reaction Scheme 3, Compound (K) may be prepared via an acid-amine condensation reaction between Compound (J) and Compound (I), and this reaction is requires a suitable condensing agent (such as HATU) and a suitable base (such as DIPEA);

[0123] Compound (K) is reacted with p-toluenesulfonyl chloride under a basic condition (such as TEA) to obtain Compound (L); and Compound (A) may be prepared via a substitution reaction between Compound (L) and Compound (E) under a basic condition (such as potassium carbonate), and according to Reaction Scheme 2, this reaction is preferably carried out at a high temperature.

##STR00047##

[0124] In the reactions shown in Reaction Scheme 4, when R.sub.1 is CH.sub.3CO— or CH.sub.3CH(CH.sub.3)CO—, Compound (E) may be prepared via an F-C acylation reaction between Compound (V) and the corresponding acyl halide or anhydride, and this reaction requires a suitable catalyst (such as aluminum trichloride).

[0125] When R.sub.1 is —CN, Compound (E) may be prepared by reacting Compound (V) with a cyanating reagent, this reaction requires a suitable cyanating reagent (such as chlorosulfonyl isocyanate), and according to Reaction Scheme 4, this reaction is preferably carried out at a low temperature.

[0126] When R.sub.1 is —CONH.sub.2, Compound (V) is reacted with a cyanating reagent to give Compound (W), this reaction requires a suitable cyanating reagent (such as chlorosulfonyl isocyanate), and according to Reaction Scheme 4, this reaction is preferably carried out at a low temperature; Compound (E) may be prepared via a hydrolysis reaction of Compound (W) under a basic condition, and this reaction requires a suitable base (such as potassium carbonate) and a suitable solvent (such as an ethanol/hydrogen peroxide mixed solvent).

[0127] When R.sub.1 is CH.sub.3SO.sub.2—, Compound (E) may be prepared via a sulfonylation reaction between Compound (V) and methanesulfonyl chloride under a basic condition, this reaction requires a suitable base (such as potassium tert-butoxide) and a suitable catalyst (such as a solution of triethylborane in tetrahydrofuran), and according to Reaction Scheme 4, this reaction is preferably carried out at a low temperature.

[0128] When R.sub.1 is CH.sub.3N(CH.sub.3)CO—, a Vilsmeier-Haack reaction of Compound (V) with POCl.sub.3 and DMF gives Compound (X). Compound (E) may be prepared by reacting Compound (X) with dimethylamine, and this reaction requires a suitable catalyst (such as sodium cyanide) and a suitable oxidant (such as manganese dioxide).

[0129] When R.sub.1 is

##STR00048##

a halogenation reaction between Compound (V) and a brominating reagent gives Compound (Y), and this reaction requires a suitable brominating reagent (such as NBS). Compound (E) may be prepared via a Suzuki coupling reaction of Compound (Y) with the corresponding boric acid or boric acid ester, this reaction requires a suitable catalyst (such as Pd(dppf)Cl.sub.2) and a suitable base (such as potassium phosphate), and according to Reaction Scheme 4, this reaction is preferably carried out at a high temperature.

[0130] When R.sub.1 is

##STR00049##

a halogenation reaction between Compound (V) and an iodinating reagent gives an intermediate compound and this reaction requires a suitable iodinating reagent (such as elemental iodine), and then the intermediate compound is reacted with Boc.sub.2O to obtain Compound (Z) and this reaction requires a suitable catalyst (such as DMAP) and a suitable base (such as TEA). Compound (E) may be prepared via a Ullmann coupling reaction between Compound (Z) and the corresponding compound with a saturated five-membered aza-ring, this reaction requires a suitable catalyst (such as cuprous iodide), a suitable ligand (such as N,N-dimethylethylenediamine) and a suitable base (such as cesium carbonate), and according to Reaction Scheme 4, this reaction is preferably carried out at a high temperature.

EXAMPLE 1

[0131] ##STR00050## ##STR00051##

Step 1:

[0132] Acetic anhydride (7.55 g, 74 mmol, 6.93 mL, 2.0 equiv.) was added dropwise into a suspension of ammonium chloride (7.92 g, 148 mmol, 5.17 mL, 4.0 equiv.) in 1,2-dichloroethane (100 mL) at 15° C. The mixture was stirred at 15° C. for 30 minutes, and a solution of Compound 1-1 (5.0 g, 37 mmol, 1.0 equiv.) in 1,2-dichloroethane (50 mL) was added into the mixture. The resulting mixture was stirred at 15° C. for 2 hours. Aluminum trichloride (9.87 g, 74 mmol, 2.0 equiv.) was added into the reaction solution, and the reaction solution was changed from heterogeneous to homogeneous. Acetic anhydride (3.78 g, 37 mmol, 3.47 mL, 1.0 equiv.) was further added into the reaction solution, and the reaction solution was stirred at 15° C. for 30 minutes. LCMS showed that the starting materials were reacted completely. The reaction solution was slowly poured into ice water (200 mL), and the resulting mixture was extracted with ethyl acetate (100 mL×2). The combined organic phases were washed with saline (100 mL), then the resulting mixture was subjected to liquid-liquid separation, and the resultant was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1 to 1:1) to obtain Compound 1-2. LCMS (ESI) m/z: 178.1 (M+1).

Step 2:

[0133] Compound 1-3 (7.11 g, 20.01 mmol, 3.0 equiv.) and potassium carbonate (4.61 g, 33.35 mmol, 5.0 equiv.) were added into a solution of Compound 1-2 (1.36 g, 6.67 mmol, 1.0 equiv.) in DMF (20 mL). Under the protection of nitrogen, the resulting mixture was heated to 100° C. and reacted for 15 hours. LCMS showed that the starting materials were not reacted completely. The reaction solution was heated to 120° C. and reacted for 2 hours. LCMS showed that the reaction was complete. Water (30 mL) was added into the reaction solution, and the resulting mixture was extracted with ethyl acetate (30 mL×3). The combined organic phases were washed with saline (30 mL) and then concentrated, and the resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1 to 3:1) to obtain Compound 1-4. LCMS (ESI) m/z: 361.1 (M+1).

Step 3:

[0134] A hydrogen chloride/ethyl acetate solution (4.0 mol/L, 20 mL, 21.57 equiv.) was added into a solution of Compound 1-4 (2.0 g, 3.71 mmol, 1.0 equiv.) in ethyl acetate (20 mL) at 0° C. After the resulting reaction solution was stirred at 15° C. for 1 hour, a large amount of solid precipitated, and LCMS showed that the reaction was complete. The reaction solution was is filtered, and the filter cake was washed with ethyl acetate (10 mL) and then dried to obtain Compound 1-5. The crude product was directly used in the next step. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.75 (s, 1H), 7.90-7.79 (m, 2H), 7.17 (dt, J=9.2, 2.6 Hz, 1H), 4.84-4.58 (m, 2H), 3.36-3.20 (m, 1H), 3.19-3.04 (m, 1H), 2.44 (d, J=2.8 Hz, 4H), 2.18-2.06 (m, 1H), 2.02 (br dd, J=7.9, 5.5 Hz, 1H), 1.88 (td, J=12.6, 7.9 Hz, 1H), 1.80-1.67 (m, 1H); LCMS (ESI) m/z: 261.1 (M+1).

Step 4:

[0135] Compound 1-6 (1.30 g, 5.05 mmol, 1.5 equiv.), HOBt (500.85 mg, 3.71 mmol, 1.1 equiv.), EDCI (710.56 mg, 3.71 mmol, 1.1 equiv.) and DIPEA (1.31 g, 10.11 mmol, 1.76 mL, 3 equiv.) were added into a solution of Compound 1-5 (1.0 g, 3.37 mmol, 1.0 equiv.) in dichloromethane (30 mL), and the resulting mixture was reacted at 15° C. for 16 hours. LCMS showed that the reaction was complete. The reaction solution was poured into water (50 mL), and the resulting mixture was extracted with dichloromethane (50 mL×3). The combined organic phases were concentrated, and the resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1 to 1:1) to obtain Compound 1-7. LCMS (ESI) m/z: 500.2 (M+1).

Step 5:

[0136] Hydrogen chloride/ethyl acetate (4.0 mol/L, 20 mL, 23.61 equiv.) was added into a solution of Compound 1-7 (1.75 g, 3.39 mmol, 1.0 equiv.) in ethyl acetate (20 mL) at 0° C. The mixture was reacted at 15° C. for 1 hour, and LCMS showed that the reaction was complete. The reaction solution was concentrated to obtain Compound 1-8, and the crude product was directly used in the next step. LCMS (ESI) m/z: 400.1 (M+1).

Step 6:

[0137] Compound 1-9 (1.05 g, 5.16 mmol, 1.5 equiv.), HOBt (511.41 mg, 3.78 mmol, 1.1 equiv.), EDCI (725.54 mg, 3.78 mmol, 1.1 equiv.) and DIPEA (1.33 g, 10.32 mmol, 1.8 mL, 3 equiv.) were added into a solution of Compound 1-8 (1.5 g, 3.44 mmol, 1.0 equiv.) in dichloromethane (30 mL), and the resulting reaction solution was reacted at 15° C. for 14 hours. LCMS showed that the reaction was complete. The reaction solution was poured into water (50 mL), the resulting mixture was extracted with dichloromethane (50 mL×3), and the combined organic phases were concentrated. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=2:1 to 1:4) to obtain Compound 1-10. LCMS (ESI) m/z: 585.3 (M+1).

Step 7:

[0138] Hydrogen chloride/ethyl acetate (4.0 mol/L, 18.57 mL, 33.41 equiv.) was added into a solution of Compound 1-10 (1.30 g, 2.22 mmol, 1.0 equiv.) in ethyl acetate (20 mL) at 0° C. The resulting mixture was reacted at 15° C. for 1 hour. LCMS showed that the reaction was complete. The reaction solution was concentrated, and the resulting residue was purified by preparative HPLC (hydrochloric acid system, mobile phase: water (0.05% hydrochloric acid)—acetonitrile, gradient: acetonitrile: 15% to 25%) to obtain the hydrochloride of Example 1. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 8.28 (s, 1H), 7.87 (dd, J=9.7, 2.4 Hz, 1H), 7.78 (dd, J=8.9, 4.2 Hz, 1H), 7.04 (dt, J=9.0, 2.4 Hz, 1H), 4.56-4.42 (m, 3H), 4.14-4.02 (m, 1H), 3.95 (q, J=6.8 Hz, 1H), 3.83 (q, J=8.4 Hz, 1H), 3.77-3.66 (m, 1H), 2.67 (s, 3H), 2.50 (s, 3H), 2.25-2.09 (m, 1H), 2.03-1.93 (m, 1H), 1.85-1.64 (m, 9H), 1.51 (d, J=7.0 Hz, 3H), 1.33-1.01 (m, 6H); LCMS (ESI) m/z: 485.2 (M+1).

EXAMPLE 2

[0139] ##STR00052## ##STR00053##

Step 1:

[0140] Acetic anhydride (6.73 g, 65.97 mmol, 6.18 mL, 2.0 equiv.) was added into a suspension of ammonium chloride (3.53 g, 65.97 mmol, 2.31 mL, 2.0 equiv.) in 1,2-dichloroethane (10 mL) at 15° C., and the mixture was stirred at 15° C. for 15 minutes. Compound 2-1 (5.0 g, 32.98 mmol, 1.0 equiv.) was added into the mixture, and the resulting mixture was stirred at 15° C. for 2 hours. Aluminum trichloride (8.80 g, 65.97 mmol, 2.0 equiv.) was added into the reaction solution, and the reaction solution was stirred at 15° C. for 30 minutes. Acetic anhydride (3.37 g, 32.98 mmol, 3.09 mL, 1.0 equiv.) was further added into the reaction solution, and the reaction solution was stirred at 15° C. for 15 minutes. LCMS showed that the starting materials were reacted completely. The reaction solution was slowly poured into ice water, and the resulting mixture was extracted with ethyl acetate (100 mL×3). The extract liquor was dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=1:1) to obtain Compound 2-2. LCMS (ESI) m/z: 194.1 (M+1).

Step 2:

[0141] Compound 2-2 (2.00 g, 10.33 mmol, 1.0 equiv.) and potassium carbonate (7.14 g, 51.65 mmol, 5.0 equiv.) were added into a solution of Compound 1-3 (7.54 g, 20.66 mmol, 2.0 equiv.) in DMF (70 mL), and the mixture was heated and stirred for 12 hours at 100° C. Water (300 mL) and ethyl acetate (300 mL) were added into the reaction solution, and the organic phase was washed with saline (100 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure to obtain Compound 2-3. LCMS (ESI) m/z: 377.0 (M+1).

Step 3:

[0142] A hydrogen chloride/dioxane solution (4.0 mol/L, 20 mL, 26.04 equiv.) was added into a solution of Compound 2-3 (2.4 g, 3.07 mmol, 1.0 equiv.) in dioxane (20 mL), and the resulting reaction solution was stirred at 15° C. for 10 hours. LCMS showed that the reaction was complete. The reaction solution was filtered, and the filter cake was washed with ethyl acetate (10 mL×3) and then dried to obtain Compound 2-4. The crude product was directly used in the next step. LCMS (ESI) m/z: 277.1 (M+1).

Step 4:

[0143] DIPEA (857.20 mg, 6.63 mmol, 1.16 mL, 3 equiv.) and HATU (1.01 g, 2.65 mmol, 1.2 equiv.) were added into a solution of Compound 1-6 (625.81 mg, 2.43 mmol, 1.1 equiv.) in DMF (5 mL), and the mixture was stirred at 15° C. for 30 minutes. Compound 2-4 (700 mg, is 2.21 mmol, 1.0 equiv., hydrochloride) was added into the reaction solution, and the reaction mixture was stirred at 15° C. for 1.5 hours. Water (30 mL) and ethyl acetate (40 mL) were added into the reaction solution. The organic phase was washed with citric acid (20 mL, 10% aqueous solution) and saline (20 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure to obtain Compound 2-5. LCMS (ESI) m/z: 516.2 (M+1).

Step 5:

[0144] Hydrogen chloride/dioxane (4.0 mol/L, 18.33 mL, 34.40 equiv.) was added into a solution of Compound 2-5 (1.10 g, 2.13 mmol, 1.0 equiv.) in dioxane (10 mL), and the mixture was reacted at 15° C. for 1.5 hours. The reaction solution was filtered, and the filter cake was washed with ethyl acetate (20 mL) and then dried to obtain Compound 2-6. LCMS (ESI) m/z: 416.2 (M+1).

Step 6:

[0145] DIPEA (326.91 mg, 2.53 mmol, 440.58 μL, 3 equiv.), HATU (384.71 mg, 1.01 mmol, 1.2 equiv.) and Compound 2-6 (500 mg, 843.16 μmol, 1.0 equiv., hydrochloride) were added into a solution of Compound 1-9 (188.50 mg, 927.48 μmol, 1.1 equiv.) in DMF (5 mL), and the reaction mixture was stirred at 15° C. for 1 hour. Water (30 mL) and ethyl acetate (20 mL) were added into the reaction solution. The organic phase was washed with citric acid (20 mL, 10% aqueous solution) and saline (20 mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure to obtain Compound 2-7. The crude product was directly used in the next step. LCMS (ESI) m/z: 601.1 (M+1).

Step 7:

[0146] Trifluoroacetic acid (3 mL) was added into a solution of Compound 2-7 (500 mg, 787.28 μmol, 1.0 equiv.) in dichloromethane (10 mL) at 0° C., and the resulting mixture was reacted at 0° C. for 1 hour. LCMS showed that the reaction was complete. The reaction solution was concentrated, and the resulting residue was purified by preparative HPLC (hydrochloric acid) to obtain the hydrochloride of Example 2. LCMS (ESI) m/z: 501.4 (M+1). .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.50 (br s, 1H), 8.88 (br d, J=5.3 Hz, 1H), 8.78 (d, J=8.2 Hz, 1H), 8.47 (s, 1H), 8.15 (d, J=2.1 Hz, 1H), 7.89 (d, J=8.8 Hz, 1H), 7.32 (dd, J=2.1, 8.7 Hz, 1H), 4.48-4.33 (m, 3H), 4.10 (br dd, J=14.8, 9.9 Hz, 1H), 3.90-3.90 (m, 1H), 3.73-3.54 (m, 2H), 2.46-2.45 (m, 1H), 2.44 (s, 3H), 2.16-2.01 (m, 1H), 1.97-1.81 (m, 1H), 1.79-1.52 (m, 9H), 1.34 (d, J=6.8 Hz, 3H), 1.27-0.87 (m, 6H).

EXAMPLE 3

[0147] ##STR00054##

[0148] Please refer to Example 1 for the preparation method of Example 3. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 8.57 (s, 1H), 8.41 (s, 1H), 7.97 (d, J=8.7 Hz, 1H), 7.55 (dd, J=8.7, 1.6 Hz, 1H), 4.64-4.54 (m, 2H), 4.48 (d, J=7.7 Hz, 1H), 4.25-4.12 (m, 1H), 3.95-3.81 (m, 2H), 3.79-3.69 (m, 1H), 2.67 (s, 3H), 2.55 (s, 3H), 2.26-2.12 (m, 1H), 2.09-1.97 (m, 1H), 1.91-1.59 (m, 9H), 1.50 (d, J=7.0 Hz, 3H), 1.36-0.97 (m, 6H); LCMS (ESI) m/z: 535.2 (M+1).

EXAMPLE 4

[0149] ##STR00055##

[0150] Please refer to Example 1 for the preparation method of Example 4. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.32 (br s, 1H), 8.85 (br d, J=5.9 Hz, 1H), 8.78 (br d, J=8.2 Hz, 1H), 8.45 (s, 1H), 7.88 (d, J=7.1 Hz, 1H), 7.31-7.20 (m, 2H), 4.48-4.36 (m, 3H), 4.14-4.04 (m, 1H), 3.92-3.81 (m, 1H), 3.73-3.64 (m, 1H), 3.63-3.56 (m, 1H), 2.49-2.45 (m, 4H), 2.17-2.02 (m, 1H), 1.96-1.84 (m, 1H), 1.81-1.56 (m, 9H), 1.34 (d, J=6.8 Hz, 3H), 1.27-0.95 (m, 6H); LCMS (ESI) m/z: 501.4 (M+1).

EXAMPLE 5

[0151] ##STR00056##

[0152] Please refer to Example 1 for the preparation method of Example 5. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.50 (br s, 1H), 8.96-8.85 (m, 1H), 8.82 (d, J=8.3 Hz, 1H), 8.45 (s, 1H), 8.15 (d, J=8.5 Hz, 1H), 8.00 (d, J=1.6 Hz, 1H), 7.24 (dd, J=8.5, 1.8 Hz, 1H), 4.47-4.35 (m, 3H), 4.08 (dd, J=14.5, 9.9 Hz, 1H), 3.89-3.80 (m, 1H), 3.74-3.66 (m, 1H), 3.64-3.56 (m, 1H), 2.46 (br s, 1H), 2.44 (s, 3H), 2.18-2.05 (m, 1H), 1.96-1.84 (m, 1H), 1.79-1.57 (m, 9H), 1.35 (d, J=6.8 Hz, 3H), 1.27-0.95 (m, 6H); LCMS (ESI) m/z: 501.4 (M+1).

EXAMPLE 6

[0153] ##STR00057##

[0154] Please refer to Example 1 for the preparation method of Example 6. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.30 (br s, 1H), 8.80 (br s, 1H), 8.59 (br d, J=7.8 Hz, 1H), 8.39 (br s, 1H), 8.19 (br d, J=7.8 Hz, 1H), 7.37-7.07 (m, 2H), 4.87-4.59 (m, 2H), 4.57-4.29 (m, 2H), 3.79 (br s, 1H), 3.65 (br s, 2H), 2.43 (br s, 7H), 2.16-1.23 (m, 13H), 1.21-0.73 (m, 5H); LCMS (ESI) m/z: is 501.3 (M+1).

EXAMPLE 7

[0155] ##STR00058##

[0156] Please refer to Example 1 for the preparation method of Example 7. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.60-9.44 (m, 1H), 8.97-8.84 (m, 1H), 8.79 (br d, J=8.2 Hz, 1H), 8.46 (s, 1H), 8.31 (d, J=1.8 Hz, 1H), 7.84 (d, J=8.8 Hz, 1H), 7.43 (dd, J=8.8, 2.0 Hz, 1H), 4.47-4.35 (m, 3H), 4.18-3.98 (m, 1H), 3.85 (br d, J=4.8 Hz, 1H), 3.73-3.64 (m, 1H), 2.47-2.42 (m, 6H), 2.17-2.02 (m, 1H), 1.98-1.85 (m, 1H), 1.81-1.52 (m, 9H), 1.34 (d, J=6.8 Hz, 3H), 1.21-0.98 (m, 5H); LCMS (ESI) m/z: 547.2 (M+1).

EXAMPLE 8

[0157] ##STR00059##

[0158] Please refer to Example 1 for the preparation method of Example 8. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.60-9.48 (m, 1H), 8.96-8.83 (m, 1H), 8.79 (d, J=8.3 Hz, 1H), 8.53 (s, 1H), 8.28 (s, 1H), 8.20 (s, 1H), 4.48-4.32 (m, 3H), 4.11 (br dd, J=13.3, 8.2 Hz, 1H), 3.83-3.77 (m, 1H), 3.73-3.65 (m, 1H), 3.64-3.52 (m, 1H), 2.47-2.43 (m, 6H), 2.21-2.03 (m, 1H), 1.96-1.84 (m, 1H), 1.77-1.72 (m, 2H), 1.67-1.55 (m, 5H), 1.34 (d, J=6.8 Hz, 3H), 1.26-0.86 (m, 6H); LCMS (ESI) m/z: 535.3 (M+1).

EXAMPLE 9

[0159] ##STR00060##

[0160] Please refer to Example 1 for the preparation method of Example 9. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 0.89-1.27 (m, 6 H) 1.34 (d, J=6.85 Hz, 3H), 1.58-1.70 (m, 5H), 1.73-1.80 (m, 2H), 1.86-1.97 (m, 1 H), 2.05-2.19 (m, 1H), 2.42-2.47 (m, 6H), 3.50-3.74 (m, 2H), 3.79-3.91 (m, 1H), 4.11 (dd, J=13.75, 8.50 Hz, 1H), 4.26-4.46 (m, 3H), 7.98 (d, J=10.15 Hz, 1H), 8.19-8.24 (m, 1H), 8.49 (s, 1H), 8.75 (d, J=8.19 Hz, 1H), 8.86 (br d, J=5.62 Hz, 1H), 9.43 (br s, 1H); LCMS (ESI) m/z: 519.3 (M+1).

EXAMPLE 10

[0161] ##STR00061##

[0162] Please refer to Example 1 for the preparation method of Example 10. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.63-9.35 (m, 1H), 8.88 (br d, J=5.3 Hz, 1H), 8.76 (br d, J=8.1 Hz, 1H), 8.55-8.43 (m, 1H), 7.73 (d, J=8.8 Hz, 1H), 7.41 (dd, J=8.4, 6.7 Hz, 1H), 4.47-4.29 (m, 3H), 4.19-4.05 (m, 1H), 3.95-3.78 (m, 1H), 3.16 (s, 3H), 2.47-2.43 (m, 5H), 2.21-2.02 (m, 1H), 1.90 (br d, J=3.3 Hz, 1H), 1.84-1.46 (m, 9H), 1.33 (br d, J=6.7 Hz, 3H), 1.22-0.91 (m, 5H); LCMS (ESI) m/z: 519.3 (M+1).

EXAMPLE 11

[0163] ##STR00062##

[0164] Please refer to Example 1 for the preparation method of Example 11. .sup.IIINNIR (400 MHz, DMSO-d.sub.6) δ 0.96-1.19 (m, 5H), 1.30-1.42 (m, 3H), 1.53-1.80 (m, 1H), 1.53-1.82 (m, 7H), 1.90 (br s, 1H), 2.02-2.17 (m, 1H), 2.41-2.48 (m, 6H), 3.57-3.63 (m, 1H), 3.83-3.91 (m, 1 H), 4.07-4.19 (m, 1H), 4.37-4.46 (m, 2H), 7.28 (br d, J=8.19 Hz, 1H), 7.97 (d, J=8.93 Hz, 1H), 8.03-8.13 (m, 1H), 8.55 (s, 1H), 8.79 (br d, J=7.95 Hz, 1H), 8.90 (br s, 1H), 9.60 (br d, J=5.01 Hz, 1H); LCMS (ESI) m/z: 551.3 (M+1).

EXAMPLE 12

[0165] ##STR00063## ##STR00064##

Step 1:

[0166] Compound 1-6 (2.54 g, 9.89 mmol, 1.0 equiv.), HATU (3.76 g, 9.89 mmol, 1.0 equiv.) and DIPEA (3.83 g, 29.66 mmol, 5.17 mL, 3.0 equiv.) were added into a solution of Compound 12-1 (1.0 g, 9.89 mmol, 0.96 mL, 1.0 equiv.) in dichloromethane (20 mL). The mixture was reacted at 30° C. for 2.0 hours. TLC (petroleum ether:ethyl acetate=1:1) detection showed that the starting materials were reacted completely. The reaction solution was poured into water (100 mL), the mixture was extracted with ethyl acetate (50 mL×2), and the combined organic phases were concentrated. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1 to 1:1) to obtain Compound 12-2. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.21 (br d, J=9.0 Hz, 1H), 4.75 (br d, J=6.4 Hz, 1H), 4.31-4.18 (m, 2H), 3.95-3.78 (m, 1H), 3.69-3.61 (m, 1H), 3.60-3.52 (m, 1H), 3.47 (td, J=10.2, 7.3 Hz, 1H), 2.12-2.03 (m, 2H), 1.96-1.80 (m, 3H), 1.69-1.51 (m, 5H), 1.41 (s, 9H), 1.22-0.97 (m, 6H); LCMS (ESI) m/z: 341.2 (M+1).

Step 2:

[0167] TEA (1.60 g, 15.86 mmol, 2.21 mL, 3.0 equiv.) and p-toluenesulfonyl chloride (1.21 g, 6.34 mmol, 1.20 equiv.) were added into a solution of Compound 12-2 (1.80 g, 5.29 mmol, 1.0 equiv.) in dichloromethane (40 mL). The resulting mixture was reacted at 30° C. for 2.0 hours. LCMS detection showed that the starting materials were not reacted completely. Water (100 mL) was added to the reaction solution, and the resulting mixture was subjected to liquid-liquid separation. The obtained organic phase was washed with saturated saline (100 mL) and concentrated to obtain Compound 12-3. The crude product was used directly in the next step. LCMS (ESI) m/z: 495.3 (M+1).

Step 3:

[0168] Please refer to the preparation method of Compound 1-4 for the preparation method of Compound 12-5. LCMS (ESI) m/z: 550.2 (M+1).

Step 4:

[0169] Please refer to the preparation method of Compound 1-8 for the preparation method of Compound 12-6. LCMS (ESI) m/z: 450.2 (M+1).

Step 5:

[0170] Please refer to the preparation method of Compound 1-10 for the preparation method of Compound 12-7. LCMS (ESI) m/z: 635.3 (M+1).

Step 6:

[0171] Please refer to Example 1 for the preparation method of the hydrochloride of Example 12. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.31 (br s, 1H), 8.87 (br d, J=8.2 Hz, 2H), 8.21 (d, J=1.6 Hz, 1H), 7.93 (d, J=8.8 Hz, 1H), 7.40 (dd, J=8.8, 1.7 Hz, 1H), 4.57-4.29 (m, 3H), 4.28-4.11 (m, 1H), 3.82 (br s, 2H), 2.60 (s, 3H), 2.11-1.93 (m, 2H), 1.81-1.51 (m, 8H), 1.44-0.90 (m, 12H); LCMS (ESI) m/z: 535.2 (M+1).

EXAMPLE 13

[0172] ##STR00065##

[0173] Please refer to Example 1 for the preparation method of the hydrochloride of Example 13. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 0.92-1.16 (m, 5H), 1.33 (br d, J=6.72 Hz, 3H), 1.47-1.68 (m, 8H), 1.91 (br d, J=6.48 Hz, 1H), 2.16-2.32 (m, 1H), 2.44 (br t, J=5.01 Hz, 3H), 2.53 (s, 3H), 2.80 (s, 3H), 3.59-3.63 (m, 1H), 3.65-3.72 (m, 1H), 3.85 (br dd, J=11.55, 6.79 Hz, 1H), 4.17-4.45 (m, 4H), 7.23 (dd, J=8.68, 1.59 Hz, 1H), 7.80 (d, J=8.80 Hz, 1H), 8.02 (d, J=1.71 Hz, 1H), 8.73 (br d, J=8.19 Hz, 1H), 8.86 (br s, 1H), 9.45 (br s, 1H); LCMS (ESI) m/z: 515.2 (M+1).

EXAMPLE 14

[0174] ##STR00066## ##STR00067##

Step 1:

[0175] Iodine (13.31 g, 52.43 mmol, 10.56 mL, 2.0 equiv.) and potassium hydroxide (5.88 g, 104.86 mmol, 4 equiv.) were added into a solution of Compound 14-1 (4 g, 26.22 mmol, 1.0 equiv.) in DMA (100 mL) at 0° C. The resulting mixture was reacted at 20° C. for 12 hours. LCMS showed that the reaction was complete. A saturated aqueous sodium sulfite solution (200 mL) was added into the reaction solution, and the resulting mixture was extracted with ethyl acetate (200 mL×2). The combined organic phases were washed with saturated saline (200 mL) and then concentrated. The resulting residue was slurried with petroleum ether (5 mL) lo to obtain Compound 14-2. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.71 (br s, 1H), 7.60 (d, J=8.8 Hz, 1H), 7.47-7.38 (m, 2H).

Step 2:

[0176] Please refer to the preparation method of Compound 1-4 for the preparation method of Compound 14-3. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.66 (d, J=8.9 Hz, 1H), 7.54-7.41 (m, 2H), 4.56-4.41 (m, 2H), 4.23-4.05 (m, 1H), 3.29-3.01 (m, 2H), 1.93-1.57 (m, 4H), 1.37 (br s, 5H), 1.11 (br s, 4H); LCMS (ESI) m/z: 484.1 (M+23).

Step 3:

[0177] Under the protection of nitrogen, zinc cyanide (0.72 g, 6.12 mmol, 0.388 mL, 0.6 equiv.), Pd.sub.2(dba).sub.3 (0.93 g, 1.02 mmol, 0.1 equiv.), zinc powder (1.33 g, 20.42 mmol, 2.0 equiv.) and DPPF (1.13 g, 2.04 mmol, 0.2 equiv.) were added into a solution of Compound 14-3 (4.80 g, 10.21 mmol, 1.0 equiv.) in DMF (100 mL). The resulting mixture was heated to 100° C. and reacted for 2 hours. LCMS showed that the reaction was complete. The reaction solution was cooled and then filtered. The filter cake was washed with ethyl acetate (50 mL) and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1 to 3:1) to obtain Compound 14-4. LCMS (ESI) m/z: 383.2 (M+23).

Step 4:

[0178] Methylmagnesium bromide (3 mol/L, 1.48 mL, 2 equiv.) was added into a solution of Compound 14-4 (0.8 g, 2.22 mmol, 1 equiv.) in tetrahydrofuran (20 mL) at 0° C., and the resulting mixture was reacted at 20° C. for 2 hours. LCMS showed that the reaction was complete. The reaction solution was slowly poured into water (100 mL), and the mixture was extracted with ethyl acetate (50 mL×2). The combined organic phases were washed with saturated saline (100 mL), then the resulting mixture was subjected to liquid-liquid separation, and the organic phase was concentrated to obtain Compound 14-5. The crude product was used directly in the next step. LCMS (ESI) m/z: 378.1 (M+1).

Step 5:

[0179] Please refer to the preparation method of Compound 1-5 for the preparation method of Compound 14-6. LCMS (ESI) m/z: 278.1 (M+1).

Step 6:

[0180] Please refer to the preparation method of Compound 1-7 for the preparation method of Compound 14-7. LCMS (ESI) m/z: 539.4 (M+23).

Step 7:

[0181] Please refer to the preparation method of Compound 1-8 for the preparation method of Compound 14-8. LCMS (ESI) m/z: 417.1 (M+1). Step 8:

[0182] Please refer to the preparation method of Compound 1-10 for the preparation method of Compound 14-9. LCMS (ESI) m/z: 624.3 (M+23).

Step 9:

[0183] Please refer to Example 1 for the preparation method of Example 14. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.06-8.79 (m, 2H), 8.70 (br d, J=8.2 Hz, 1H), 8.17-8.10 (m, 1H), 7.97 (br d, J=8.9 Hz, 1H), 7.56 (br d, J=8.7 Hz, 1H), 4.71 (br dd, J=13.2, 3.8 Hz, 1H), 4.58-4.44 (m, 2H), 4.38 (br t, J=7.6 Hz, 1H), 3.85 (br d, J=5.1 Hz, 2H), 2.62 (s, 3H), 1.92-1.74 (m, 5H), 1.72-1.49 (m, 8H), 1.31 (br d, J=6.7 Hz, 3H), 1.13 (br d, J=13.7 Hz, 4H), 1.04-0.91 (m, 2H); LCMS (ESI) m/z: 502.1 (M+1).

EXAMPLE 15

[0184] ##STR00068##

[0185] Please refer to Example 14 for the preparation method of Example 15. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.56 (br s, 1H), 8.87 (br s, 1H), 8.80-8.60 (m, 1H), 8.14-7.94 (m, 1H), 7.85-7.68 (m, 1H), 7.58-7.40 (m, 1H), 4.78-4.30 (m, 4H), 3.96-3.74 (m, 1H), 3.64-3.58 (m, 2H), 2.61 (s, 1H), 2.47-2.38 (m, 3H), 1.94-1.52 (m, 11H), 1.39-1.26 (m, 3H), 1.24-0.87 (m, 6H); LCMS (ESI) m/z: 486.3 (M+1).

EXAMPLE 16

[0186] ##STR00069## ##STR00070##

Step 1:

[0187] Please refer to the preparation method of Compound 1-4 for the preparation method of Compound 16-2. LCMS (ESI) m/z: 336.2 (M+1).

Step 2:

[0188] Compound 16-3 (997.86 mg, 7.05 mmol, 612.18 μL, 1.5 equiv.) was added into a solution of Compound 16-2 in DMF (20 mL) at −20° C., and the resulting mixture was stirred at is 0° C. for 2 hours. TLC showed that the starting materials were reacted completely. 1.5 mL of Compound 16-3 (2.44 g, 17.28 mmol, 1.5 mL, 3.68 equiv.) was additionally added into the above reaction solution at −20° C., and the resulting mixture was stirred at 0° C. for 0.5 hour. LCMS showed that the starting materials were reacted completely. Water (50 mL) was added into the reaction mixture, and the resulting mixture was filtered. The filter cake was washed with water (20 mL×2) and then dried in vacuum. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=50:1 to 10:1) to obtain Compound 16-3. .sup.1 H NMR (400 MHz, DMSO-d.sub.6) δ 0.98 (br s, 5H), 1.14-1.31 (m, 4H), 1.80 (br d, J=17.85 Hz, 4H), 3.17-3.31 (m, 2H), 4.12-4.30 (m, 2H), 4.35-4.44 (m, 1H), 8.24-8.39 (m, 1H), 8.42-8.57 (m, 2H); LCMS (ESI) m/z: 361.1 (M+1).

Step 3:

[0189] Please refer to the preparation method of Compound 14-5 for the preparation method of Compound 16-5. LCMS (ESI) m/z: 378.2 (M+1). Step 4:

[0190] Please refer to the preparation method of Compound 1-5 for the preparation method of Compound 16-6. LCMS (ESI) m/z: 278.0 (M+1).

Step 5:

[0191] Please refer to the preparation method of Compound 1-7 for the preparation method of Compound 16-7. LCMS (ESI) m/z: 539.4 (M+23). Step 6:

[0192] Please refer to the preparation method of Compound 1-8 for the preparation method of Compound 16-8. LCMS (ESI) m/z: 417.3 (M+1).

Step 7:

[0193] Please refer to the preparation method of Compound 1-10 for the preparation method of Compound 16-9. LCMS (ESI) m/z: 624.1 (M+23).

Step 8:

[0194] Please refer to Example 1 for the preparation method of Example 16. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 1.02-1.14 (m, 6H), 1.35-1.49 (m, 2H), 1.58 (br s, 4 H), 1.66-1.80 (m, 4H), 1.87 (br d, J=6.72 Hz, 2H), 2.09-2.20 (m, 3H), 2.46 (s, 3H), 2.93 (dt, J=13.57, 6.79 Hz, 1H), 3.54 (br d, J=9.90 Hz, 2H), 3.62-3.70 (m, 1H), 4.21-4.41 (m, 2H), 4.48 (br dd, J=13.51, 7.03 Hz, 1H), 4.68 (br d, J=3.18 Hz, 1H), 7.80 (br d, J=8.80 Hz, 1H), 8.32-8.40 (m, 1H), 8.40-8.47 (m, 1H), 8.55-8.66 (m, 1H); LCMS (ESI) m/z: 502.1 (M+1).

EXAMPLE 17

[0195] ##STR00071##

[0196] Please refer to Example 16 and Example 1 for the preparation method of Example 17. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 8.53 (br d, J=7.6 Hz, 1H), 8.06 (s, 1H), 7.88 (dd, J=8.9, 4.2 Hz, 1H), 7.32 (dd, J=8.8, 2.3 Hz, 1H), 7.14 (dt, J=9.1, 2.3 Hz, 1H), 4.57-4.45 (m, 3H), 4.21-4.08 (m, 1H), 3.92 (q, J=6.8 Hz, 1H), 3.86-3.78 (m, 1H), 3.76-3.65 (m, 1H), 2.67 (s, 3H), 2.22-2.07 (m, 1H), 2.04-1.96 (m, 1H), 1.83-1.63 (m, 8H), 1.50 (d, J=6.8 Hz, 3H), 1.3-1.02 (m, 6H); LCMS (ESI) m/z: 468.2 (M+1).

EXAMPLE 18

[0197] ##STR00072##

[0198] Please refer to Example 16 and Example 1 for the preparation method of Example 18. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.60-9.41 (m, 1H), 8.86 (br d, J=4.6 Hz, 1H), 8.69 (d, J=8.3 Hz, 1H), 8.37 (s, 1H), 7.94 (d, J=8.8 Hz, 1H), 7.66 (d, J=1.8 Hz, 1H), 7.39 (dd, J=8.8, 2.0 Hz, 1H), 4.41 (br d, J=2.8 Hz, 1H), 4.38 (br d, J=6.5 Hz, 1H), 4.23-4.12 (m, 1H), 3.88-3.79 (m, 1H), 3.70-3.53 (m, 2H), 2.44 (br t, J=5.1 Hz, 3H), 2.15-2.01 (m, 1H), 1.90 (br d, J=4.3 Hz, 1H), 1.75-1.52 (m, 8H), 1.32 (d, J=6.8 Hz, 3H), 1.25-0.89 (m, 6H); LCMS (ESI) m/z: 484..4 (M+1).

EXAMPLE 19

[0199] ##STR00073##

[0200] Please refer to Example 14 for the preparation method of Example 19. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.93 (d, J=9.0 Hz, 1H), 7.83 (d, J=1.1 Hz, 1H), 7.54 (dd, J=8.9, 1.5 Hz, 1H), 4.77-4.70 (m, 1H), 4.68-4.61 (m, 1H), 4.57 (br s, 1H), 4.48 (br d, J=7.0 Hz, 1H), 3.88 (q, J=7.0 Hz, 1H), 3.84-3.74 (m, 1H), 3.65-3.54 (m, 1H), 2.68-2.62 (m, 3H), 1.97-1.85 (m, 3H), 1.83-1.66 (m, 7H), 1.47 (d, J=7.0 Hz, 3H), 1.37-1.00 (m, 5H); LCMS (ESI) m/z: 485.1 (M+1).

EXAMPLE 20

[0201] ##STR00074## ##STR00075##

Step 1:

[0202] Under the protection of nitrogen, potassium tert-butoxide (0.91 g, 8.14 mmol, 1.1 equiv.) was added into a solution of Compound 1-1 (1.0 g, 7.40 mmol, 1.0 equiv.) in tetrahydrofuran (20 mL). After the mixture was reacted at 15° C. for 0.5 hour, a solution of triethylborane in tetrahydrofuran (1 mol/L, 8.14 mL, 1.1 equiv.) was slowly added into the reaction solution. The reaction solution was further reacted at 15° C. for 0.5 hour, and methanesulfonyl chloride (0.93 g, 8.14 mmol, 0.63 mL, 1.1 equiv.) was then added into the reaction solution. The reaction solution was reacted at −15° C. for 10 hours. LCMS showed that the reaction was complete. A saturated aqueous ammonium chloride solution (100 mL) was added into the reaction system, is the resulting mixture was extracted with ethyl acetate (100 mL×2), and the combined organic phases were concentrated to obtain Compound 20-1. LCMS (ESI) m/z: 214.0 (M+1).

Step 2:

[0203] Please refer to the preparation method of Compound 1-4 for the preparation method of Compound 20-2. LCMS (ESI) m/z: 397.1 (M+1).

Step 3:

[0204] Please refer to the preparation method of Compound 1-5 for the preparation method of Compound 20-3. LCMS (ESI) m/z: 297.1 (M+1).

Step 4:

[0205] Please refer to the preparation method of Compound 1-7 for the preparation method of Compound 20-4. LCMS (ESI) m/z: 558.1 (M+23).

Step 5:

[0206] Please refer to the preparation method of Compound 1-8 for the preparation method of Compound 20-5.

Step 6:

[0207] Please refer to the preparation method of Compound 1-10 for the preparation method of Compound 20-6. LCMS (ESI) m/z: 643.4 (M+23).

Step 7:

[0208] Please refer to Example 1 for the preparation method of Example 20. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.61-9.37 (m, 1H), 8.96-8.83 (m, 1H), 8.79 (d, J=8.2 Hz, 1H), 8.23-8.19 (m, 1H), 7.98 (dd, J=9.2, 4.4 Hz, 1H), 7.52 (dd, J=9.4, 2.5 Hz, 1H), 7.26 (dt, J=9.2, 2.5 Hz, 1H), 4.51-4.32 (m, 3H), 4.15 (dd, J=13.2, 8.6 Hz, 1H), 3.98-3.79 (m, 1H), 3.20 (s, 3H), 2.46 (t, J=5.3 Hz, 3H), 2.08 (td, J=12.2, 8.7 Hz, 1H), 1.95-1.82 (m, 1H), 1.77-1.49 (m, 9H), 1.37-1.30 (m, 3H), 1.26-0.91 (m, 6H); LCMS (ESI) m/z: 521.3 (M+1).

EXAMPLE 21

[0209] ##STR00076##

[0210] Please refer to Example 20 for the preparation method of Example 21. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 8.03 (s, 1H), 7.91-7.80 (m, 2H), 7.32 (dd, J=8.7, 1.9 Hz, 1H), 4.59-4.43 (m, 2H), 4.21-4.10 (m, 1H), 3.94 (q, J=6.8 Hz, 1H), 3.82 (q, J=8.5 Hz, 1H), 3.74-3.63 (m, 1H), 3.18 (s, 3H), 2.67 (s, 3H), 2.16-2.02 (m, 1H), 2.01-1.94 (m, 1H), 1.85-1.72 (m, 6H), 1.71-1.59 (m, 2H), 1.50 (d, J=7.0 Hz, 3H), 1.34-1.01 (m, 6H); LCMS (ESI) m/z: 537.1 (M+1).

EXAMPLE 22

[0211] ##STR00077##

Step 1:

[0212] Potassium carbonate (85.21 mg, 616.53 μmol, 2.0 equiv.) and hydrogen peroxide (9.44 g, 83.26 mmol, 8 mL, concentration: 30%, 270.09 equiv.) were added into a solution of Compound 17-8 (175 mg, 308.27 μmol, 1.0 equiv.) in ethanol (8 mL), and the resulting reaction solution was reacted at 50° C. for 1 hour. LCMS showed that the reaction was complete. Water (40 mL) was added into the reaction solution, and the resulting mixture was extracted with ethyl acetate (30 mL). The combined organic phases were washed with saturated saline (20 mL), dried over anhydrous sodium sulfate, and then filtered. The filtrate was lo concentrated under reduced pressure to obtain Compound 22-2. LCMS (ESI) m/z: 586.6 (M+1).

Step 2:

[0213] Please refer to Example 1 for the preparation method of Example 22. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.48 (br s, 1H), 8.88 (br s, 1H), 8.81 (br d, J=7.9 Hz, 1H), 8.16 (s, 1H), 7.87-7.78 (m, 2H), 7.13-7.06 (m, 1H), 4.44-4.36 (m, 2H), 4.32 (br d, J=4.1 Hz, 1H), 4.03 (br dd, J=13.5, 9.6 Hz, 1H), 3.86 (br d, J=4.9 Hz, 2H), 3.63-3.53 (m, 1H), 2.44 (br s, 1H), 2.15-2.02 (m, 1H), 1.91 (br s, 1H), 1.77-1.58 (m, 9H), 1.35 (br d, J=6.8 Hz, 3H), 1.21-0.99 (m, 6H); LCMS (ESI) m/z: 486.5 (M+1).

EXAMPLE 23

[0214] ##STR00078## ##STR00079##

Step 1:

[0215] POCl.sub.3 (3.96 g, 25.83 mmol, 2.40 mL, 1.31 equiv.) was slowly added dropwise into a solution of DMF (8.55 g, 116.97 mmol, 9 mL, 5.91 equiv.) at 0° C. within 30 minutes. A solution of Compound 23-1 (3.0 g, 19.79 mmol, 1.0 equiv.) in DMF (3 mL) was added dropwise into the mixed solution, and the resulting mixture was stirred at 25° C. for 1 hour. TLC (petroleum ether:ethyl acetate=3:1) and LCMS showed that the starting materials were reacted completely. The reaction solution was slowly poured into water (150 mL), the pH of the mixed solution was adjusted to 9 with a 10% NaOH solution, and then the resulting mixture was extracted with ethyl acetate (200 mL×2). The combined organic phases were washed with saturated saline (100 mL×2), dried over anhydrous sodium sulfate and then concentrated to obtain Compound 23-2. .sup.1 H NMR (400 MHz, DMSO-d.sub.6) δ 7.28 (dd, J=8.66, 2.13 Hz, 1H), 7.54 (d, J=8.66 Hz, 1H), 8.06 (d, J=2.01 Hz, 1H), 8.36 (s, 1H), 9.93 (s, 1H), 12.29 (br s, 1H); LCMS (ESI) m/z: 180.1 (M+1).

Step 2:

[0216] Compound 23-3 (2 mol/L, 33.41 mL, 4.0 equiv.) was added into a solution of Compound 23-2 (3 g, 16.70 mmol, 1.0 equiv.) and sodium cyanide (163.72 mg, 3.34 mmol, 0.2 equiv.) in DMF (30 mL), and the resulting mixture was stirred at 30° C. for 10 minutes. Manganese dioxide (36.30 g, 417.59 mmol, 25.0 equiv.) was added in portions into the mixture, and the resulting mixture was further stirred at 30° C. for 14 hours. LCMS showed that the starting materials were reacted completely. The reaction mixture was filtered, and the filter cake was washed with ethyl acetate (100 mL×2). The combined organic phases were successively washed with a saturated ferrous sulfate solution (50 mL×2) and saline (100 mL), dried over anhydrous sodium sulfate and then concentrated to obtain Compound 23-4. The crude product was used directly in the next step. LCMS (ESI) m/z: 223.2 (M+1).

Step 3:

[0217] Please refer to the preparation method of Compound 1-4 for the preparation method of Compound 23-5. LCMS (ESI) m/z: 406.0 (M+1).

Step 4:

[0218] Please refer to the preparation method of Compound 1-5 for the preparation method of Compound 23-6. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 1.62-1.77 (m, 1H), 1.80-1.92 (m, 1H), 1.94-2.04 (m, 1H), 2.06-2.17 (m, 1H), 3.04-3.15 (m, 6H) 3.20-3.31 (m, 1H), 3.86 (br s, 1H), 4.52-4.56 (m, 1H), 4.61 (br dd, J=14.87, 5.08 Hz, 2H), 4.69-4.77 (m, 1H), 4.69-4.77 (m, 1H), 7.26 (dd, J=8.72, 2.07 Hz, 1H), 7.79 (d, J=8.78 Hz, 1H), 7.90 (d, J=2.01 Hz, 1H), 8.26 (s, 1H), 9.35 (br s, 1H), 10.04 (br s, 1H); LCMS (ESI) m/z: 306.1(M+1).

Step 5:

[0219] Please refer to the preparation method of Compound 1-7 for the preparation method of Compound 23-7. LCMS (ESI) m/z: 545.4 (M+1)

Step 6:

[0220] Please refer to the preparation method of Compound 1-8 for the preparation method of Compound 23-8. LCMS (ESI) m/z: 445.0 (M+1).

Step 7:

[0221] Please refer to the preparation method of Compound 1-10 for the preparation method of Compound 23-9. LCMS (ESI) m/z: 630.3 (M+1).

Step 8:

[0222] Please refer to Example 1 for the preparation method of Example 23. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 1.00-1.30 (m, 5H), 1.47-1.54 (m, 3H), 1.61 (br d, J=12.23 Hz, 1H), 1.65-1.87 (m, 7H), 1.99 (br s, 1H), 2.06-2.23 (m, 1H), 2.64-2.71 (m, 3H), 3.1-3.30 (m, 6H), 3.72 (br d, J=6.48 Hz, 1H), 3.83 (q, J=8.48 Hz, 1H), 3.90-4.02 (m, 1H), 4.09-4.20 (m, 1H), 4.46 (br d, J=7.46 Hz, 1H), 4.51-4.62 (m, 2H), 7.24-7.30 (m, 1H), 7.74 (s, 1H), 7.80 (d, J=9.05 Hz, 1H), 7.92 (br s, 1H); LCMS (ESI) m/z: 530.3(M+1).

EXAMPLE 24

[0223] ##STR00080##

[0224] Please refer to Example 1 for the preparation method of Example 24. .sup.1H NMR (400MIlz, CD.sub.3OD) δ 8.36-8.23 (m, 2H), 7.79 (d, J=8.8 Hz, 1H), 7.27 (dd, J=8.8, 2.0 Hz, 1H), 4.60-4.44 (m, 3H), 4.21-4.05 (m, 1H), 3.97-3.77 (m, 2H), 3.71 (qd, J=10.0, 4.0 Hz, 1H), 3.44 (spt, J=6.8 Hz, 1H), 2.67 (s, 3H), 2.23-2.07 (m, 1H), 2.06-1.96 (m, 1H), 1.85-1.64 (m, 8H), 1.50 (d, J=7.1 Hz, 3H), 1.27-1.05 (m, 10H); LCMS (ESI) m/z: 551.3 (M+23).

EXAMPLE 25

[0225] ##STR00081## ##STR00082##

Step 1:

[0226] NBS (276.58 mg, 1.55 mmol, 1.05 equiv.) was added in portions into a solution of Compound 1-1 (0.2 g, 1.48 mmol, 1.0 equiv.) in DMF (2 mL), and the resulting mixture was stirred at 15° C. for 1 hour. TLC (petroleum ether:ethyl acetate=3:1) showed that the starting materials were reacted completely, and LCMS showed that a product was formed. A saturated sodium sulfite solution (2 mL) was added into the reaction solution, and the mixture was extracted with ethyl acetate (2 mL×3). The combined organic phases were concentrated to obtain Compound 25-1, and the crude product was directly used in the next step. LCMS (ESI) m/z: 211.9 (M-1).

Step 2:

[0227] Under the protection of nitrogen, Compound 25-2 (1.91 g, 14.02 mmol, 2.0 equiv.), potassium phosphate (2.98 g, 14.02 mmol, 2.0 equiv.) and Pd(dppf)Cl.sub.2 (512.80 mg, 700.82 μmol, 0.1 equiv.) were added into a mixed solution of Compound 25-1 (1.5 g, 7.01 mmol, 1.0 equiv.) in tetrahydrofuran (18 mL) and water (3.0 mL), and the resulting mixture was heated to 80° C. and stirred for 16 hours. LCMS showed that the starting materials were reacted completely. The reaction solution was dried over anhydrous sodium sulfate and then filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=1:0 to 5:1) to obtain Compound 25-3. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.07-7.86 (m, 1H), 7.52 (d, J=2.4 Hz, 1H), 7.46-7.40 (m, 1H), 7.37-7.30 (m, 1H), 7.26-7.21 (m, 2H), 7.13-7.05 (m, 2H), 6.99 (dt, J=9.1, 2.3, 1H), 2.29 (s, 3H); LCMS (ESI) m/z: 224.0 (M-1).

Step 3:

[0228] Please refer to the preparation method of Compound 1-4 for the preparation method of Compound 25-4. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.43-7.24 (m, 5H), 7.22-7.13 (m, 1H), 7.13-7.07 (m, 1H), 7.05-6.97 (m, 1H), 4.49-4.36 (m, 1H), 4.31-4.24 (m, 1H), 3.50-3.33 (m, 1H), 3.26-3.11 (m, 1H), 2.35 (s, 3H), 1.95-1.85 (m, 1H), 1.83-1.71 (m, 2H), 1.59-1.46 (m, 11H); LCMS (ESI) m/z: 431.3 (M+23).

Step 4:

[0229] Please refer to the preparation method of Compound 1-5 for the preparation method of Compound 25-5. LCMS (ESI) m/z: 309.2 (M+1).

Step 5:

[0230] Please refer to the preparation method of Compound 1-7 for the preparation method of

[0231] Compound 25-6. LCMS (ESI) m/z: 548.1 (M+1).

Step 6:

[0232] Please refer to the preparation method of Compound 1-8 for the preparation method of Compound 25-7. LCMS (ESI) m/z: 448.2 (M+1).

Step 7:

[0233] Please refer to the preparation method of Compound 1-10 for the preparation method of Compound 25-8. LCMS (ESI) m/z: 633 (M+1).

Step 8:

[0234] Please refer to Example 1 for the preparation method of Example 25. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.70 (dd, J=8.9, 4.1 Hz, 1H), 7.31 (dd, J=4.7, 3.2 Hz, 2H), 7.27 (s, 1H), 7.25-7.19 (m, 2H), 7.03-6.94 (m, 2H), 4.62-4.47 (m, 3H), 4.21-4.07 (m, 1H), 3.97-3.88 (m, 1H), 3.86-3.74 (m, 1H), 3.70-3.59 (m, 1H), 2.68 (s, 3H), 2.28 (s, 3H), 2.09-1.57 (m, 11H), 1.52 (d, J=6.9 Hz, 3H), 1.39-1.02 (m, 6H); LCMS (ESI) m/z: 533.2 (M+1).

EXAMPLE 26

[0235] ##STR00083##

[0236] Please refer to Example 25 for the preparation method of Example 26. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 8.23-8.14 (m, 1H), 8.03-7.93 (m, 1H), 7.88 (dd, J=8.9, 4.3 Hz, 1H), 7.39 (br d, J=9.4 Hz, 1H), 7.12 (dt, J=9.1, 2.4 Hz, 1H), 6.94-6.85 (m, 1H), 4.65-4.56 (m, 2H), 4.48 (d, J=7.5 Hz, 1H), 4.25-4.08 (m, 4H), 4.00-3.91 (m, 1H), 3.90-3.80 (m, 1H), 3.79-3.68 (m, 1H), 2.67 (s, 3H), 2.30-2.13 (m, 1H), 2.05-1.95 (m, 1H), 1.90-1.60 (m, 8H), 1.51 (d, J=7.0 Hz, 3H), 1.38-1.01 (m, 6H); LCMS (ESI) m/z: 523.3 (M+1).

EXAMPLE 27

[0237] ##STR00084##

[0238] Please refer to Example 25 for the preparation method of Example 27. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.83-7.95 (m, 2H), 7.82 (s, 1H), 7.53-7.59 (m, 1H), 7.47-7.53 (m, 1H), 7.29 (dd, J=8.78, 2.01 Hz, 1H), 6.48 (d, J=1.76 Hz, 1H), 4.34-4.53 (m, 3H), 4.10 (dd, J=13.05, 8.03 Hz, 1H), 3.86-3.89 (m, 3H), 3.61 (br d, J=8.03 Hz, 2H), 2.97 (q, J=6.69 Hz, 1H), 2.17 (s, 3H), 1.98-2.07 (m, 1H), 1.78-1.92 (m, 2H), 1.51-1.77 (m, 9H), 1.14 (br s, 1H), 1.09 (d, J=6.78 Hz, 4H), 0.89-1.04 (m, 2H); LCMS (ESI) m/z: 539.3 (M+1).

EXAMPLE 28

[0239] ##STR00085## ##STR00086##

Step 1:

[0240] A solution of iodine (1.67 g, 6.60 mmol, 1.0 equiv.) in DMF (20 mL) and potassium hydroxide (0.92 g, 16.49 mmol, 2.5 equiv.) were added into a solution of Compound 23-1 (1.0 g, 6.60 mmol, 1.0 equiv.) in DMF (20 mL) at 20° C., and the resulting mixture was stirred and reacted at 20° C. for 1 hour. TLC (petroleum ether:ethyl acetate=3:1) detection showed that the starting materials were reacted completely. The reaction solution was slowly poured into a saturated aqueous sodium sulfite solution (100 mL), and then the mixture was extracted with ethyl acetate (100 mL). The combined organic phases were washed with saturated saline (100 mL) and then concentrated to obtain Compound 28-1. The crude product was used directly in the next step. LCMS (ESI) m/z: 277.9 (M+1).

Step 2:

[0241] Boc.sub.2O (1.42 g, 6.49 mmol, 1.49 mL, 1.2 equiv.), TEA (1.64 g, 16.22 mmol, 1 equiv.) and DMAP (66 mg, 0.54 mmol, 0.1 equiv.) were added into a solution of Compound 28-1 (1.5 g, 5.41 mmol, 1.0 equiv.) in dichloromethane (50 mL). The resulting mixture was stirred and reacted at 20° C. for 10 hours. TLC (petroleum ether:ethyl acetate=10:1) detection showed that the starting materials were reacted completely. The reaction mixture was concentrated, and the resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=20:1 to 10:1) to obtain Compound 28-2. .sup.1 H NMR (400MHz, DMSO-d.sub.6) δ 8.01 (d, J=8.8 Hz, 1H), 7.91 (s, 1H), 7.39 (dd, J=8.8, 2.0 Hz, 1H), 7.30 (d, J=1.8 Hz, 1H), 1.61 (s, 9H); LCMS (ESI) m/z: 378.0(M+1).

Step 3:

[0242] Under the protection of nitrogen, Compound 28-3 (1.01 g, 11.92 mmol, 0.91 mL, 3.0 equiv.), cuprous iodide (0.37 mg, 1.99 mmol, 0.5 equiv.), cesium carbonate (3.88 g, 11.92 mmol, 3.0 equiv.), and N,N-dimethylethylenediamine (0.35 g, 3.97 mmol, 1.0 equiv.) were added into a solution of Compound 28-2 (1.5 g, 3.97 mmol, 1.0 equiv.) in dioxane (40 mL). The reaction solution was heated to 80° C. and reacted for 2.0 hours. LCMS showed that the starting materials were reacted completely. The reaction solution was cooled to 20° C. and filtered. The filter cake was washed with ethyl acetate (50 mL), and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1 to 1:1) to obtain Compound 28-4. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.97 (br s, 1H), 7.53 (d, J=1.8 Hz, 1H), 7.19-7.01 (m, 3H), 3.92 (t, J=7.0 Hz, 2H), 2.64 (t, J=8.2 Hz, 2H), 2.32-2.24 (m, 2H); LCMS (ESI) m/z: 234.1 (M+1).

Step 4:

[0243] Please refer to the preparation method of Compound 1-4 for the preparation method of Compound 28-5. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.61 (br d, J=6.1 Hz, 1H), 7.48 (br d, J=8.7 Hz, 1H), 7.44-7.38 (m, 1H), 7.32 (br d, J=8.7 Hz, 1H), 7.15 (dd, J=8.8, 1.9 Hz, 1H), 4.22-4.13 (m, 1H), 3.98 (br t, J=7.0 Hz, 2H), 3.45-3.15 (m, 2H), 2.95 (s, 1H), 2.87 (s, 1H), 2.59 (t, J=8.1 Hz, 2H), 2.30-2.16 (m, 2H), 1.93-1.65 (m, 4H), 1.50 (s, 10H); LCMS (ESI) m/z: 418.2 (M+1).

Step 5:

[0244] Please refer to the preparation method of Compound 1-5 for the preparation method of Compound 28-6. LCMS (ESI) m/z: 318.1 (M+1).

Step 6:

[0245] Please refer to the preparation method of Compound 1-7 for the preparation method of Compound 28-7. LCMS (ESI) m/z: 557.3 (M+1).

Step 7:

[0246] Please refer to the preparation method of Compound 1-8 for the preparation method of Compound 28-8. LCMS (ESI) m/z: 457.2 (M+1).

Step 8:

[0247] Please refer to the preparation method of Compound 1-10 for the preparation method of

[0248] Compound 28-9. LCMS (ESI) m/z: 642.3 (M+1).

Step 9:

[0249] Please refer to Example 1 for the preparation method of Example 28. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.41 (br s, 1H), 9.08-8.72 (m, 2H), 7.84-7.49 (m, 3H), 7.26-7.09 (m, 1H), 4.51-4.23 (m, 3H), 4.10-3.79 (m, 4H), 3.72-3.51 (m, 1H), 2.46 (br s, 6H), 2.14 (br d, J=6.0 Hz, 2H), 1.95 (br s, 1H), 1.90-1.54 (m, 9H), 1.35 (br d, J=6.2 Hz, 3H), 1.29-0.94 (m, 6H); LCMS (ESI) m/z: 542.3 (M+1).

EXAMPLE 29

[0250] ##STR00087##

[0251] Please refer to Example 28 for the preparation method of Example 29. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.67 (d, J=8.8 Hz, 1H), 7.57 (d, J=1.8 Hz, 1H), 7.31 (s, 1H), 7.15 (dd, J=8.7, 1.9 Hz, 1H), 4.59-4.38 (m, 4H), 4.08-3.98 (m, 1H), 3.96-3.90 (m, 1H), 3.88-3.81 (m, 2H), 3.81-3.74 (m, 1H), 3.72-3.64 (m, 1H), 3.61-3.53 (m, 2H), 2.89 (s, 4H), 2.67 (s, 3H), 1.93 (br d, J=9.7 Hz, 2H), 1.88-1.66 (m, 9H), 1.52-1.49 (m, 3H), 1.36-1.08 (m, 6H); LCMS (ESI) m/z: 557.3 (M+1).

EXAMPLE 30

[0252] ##STR00088##

[0253] Please refer to Example 28 for the preparation method of Example 30. .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.70 (d, J=8.8 Hz, 1H), 7.63 (d, J=2.0 Hz, 1H), 7.45 (s, 1H), 7.21-7.14 (m, 1H), 4.60-4.55 (m, 2H), 4.54-4.43 (m, 3H), 4.12 (dt, J=8.1, 3.5 Hz, 2H), 4.07-3.98 (m, 1H), 3.97-3.89 (m, 1H), 3.84-3.75 (m, 1H), 3.73-3.64 (m, 1H), 2.67 (s, 3H), 2.00-1.89 (m, 2H), 1.87-1.76 (m, 6H), 1.71 (br d, J=10.8 Hz, 2H), 1.51 (d, J=7.0 Hz, 3H), 1.36-1.06 (m, 6H); LCMS (ESI) m/z: 544.3 (M+1).

EXAMPLE 31

[0254] ##STR00089## ##STR00090##

Step 1:

[0255] Potassium carbonate (17.93 g, 129.73 mmol, 3.0 equiv.) and methyl iodide (9.21 g, 64.87 mmol, 4.04 mL, 1.5 equiv.) were added into a solution of Compound 31-1 (10 g, 43.24 mmol, 1.0 equiv.) in DMF (100 mL), and the resulting mixture was reacted at 15° C. for 5 hours. TLC (petroleum ether:ethyl acetate=1:1) showed that the reaction was complete. Water (100 mL) was added into the reaction solution, and the resulting mixture was extracted with ethyl acetate (100 mL×2). The combined organic phases were washed with saturated saline (100 mL×3) and then concentrated to obtain Compound 31-2. The crude product was used directly in the next step.

Step 2:

[0256] Under the protection of nitrogen, phenol (3.8 g, 40.36 mmol, 3.55 mL, 1.1 equiv.), triphenylphosphine (10.59 g, 40.36 mmol, 1.1 equiv.) and DIAD (8.16 g, 40.36 mmol, 7.85 mL, 1.1 equiv.) were added into a solution of Compound 31-2 (9.0 g, 36.69 mmol, 1.0 equiv.) in tetrahydrofuran (150 mL). The resulting mixture was stirred at 15° C. for 12 hours. LCMS showed that the reaction was complete. The reaction solution was concentrated, water (100 mL) and ethyl acetate (200 mL) were added into the resulting residue, and the resulting mixture was subjected to liquid-liquid separation. The organic phase was washed with saturated saline (100 mL×2) and then concentrated to obtain Compound 31-3. The crude product was used directly in the next step. LCMS (ESI) m/z: 322.2 (M+1).

Step 3:

[0257] Lithium aluminum hydride (1.59 g, 42.01 mmol, 1.5 equiv.) was added into a solution of Compound 31-3 (9 g, 28.01 mmol, 1.0 equiv.) in tetrahydrofuran (100 mL) at 0° C., and the resulting mixture was reacted at 15° C. for 2 hours. LCMS showed that the reaction was complete. To the reaction solution, water (3 mL), a 30% sodium hydroxide solution (6 mL) and water (3 mL) were successively added dropwise to quench the reaction. The resulting mixture was filtered, the filter cake was washed with ethyl acetate (100 mL), and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:to 2:1) to obtain Compound 31-4. LCMS (ESI) m/z: 316.2 (M+23).

Step 4:

[0258] Pyridine (4.04 g, 51.13 mmol, 4.13 mL, 3.0 equiv.) and p-toluenesulfonyl chloride (6.50 g, 34.09 mmol, 2.0 equiv.) were added into a solution of Compound 31-4 (5 g, 17.04 mmol, 1.0 equiv.) in dichloromethane (150 mL) at 0° C., and the resulting mixture was stirred at 15° C. for 10 hours. LCMS showed that the reaction was complete. The reaction solution was concentrated, and the resulting residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=20:1 to 3:1) to obtain Compound 31-5. LCMS (ESI) m/z: 470.2 (M+23).

Step 5:

[0259] Please refer to the preparation method of Compound 1-4 for the preparation method of Compound 31-7. LCMS (ESI) m/z: 453.1 (M+1).

Step 6:

[0260] Please refer to the preparation method of Compound 1-5 for the preparation method of Compound 31-8.

Step 7:

[0261] Please refer to the preparation method of Compound 1-7 for the preparation method of Compound 31-9. LCMS (ESI) m/z: 592.1 (M+1).

Step 8:

[0262] Please refer to the preparation method of Compound 1-8 for the preparation method of Compound 31-10.

Step 9:

[0263] Please refer to the preparation method of Compound 1-10 for the preparation method of Compound 31-11. LCMS (ESI) m/z: 677.2 (M+1).

Step 10:

[0264] Please refer to Example 1 for the preparation method of Example 31. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 9.60-9.34 (m, 1H), 8.98-8.83 (m, 1H), 8.80 (d, J=7.8 Hz, 1H), 7.98 (s, 1H), 7.83 (dd, J=9.8, 2.6 Hz, 1H), 7.73 (dd, J=9.0, 4.5 Hz, 1H), 7.44-7.34 (m, 2H), 7.15-7.08 (m, 3H), 7.07-7.01 (m, 1H), 5.22 (br s, 1H), 4.73-4.56 (m, 2H), 4.37 (t, J=7.6 Hz, 1H), 4.33-4.22 (m, 1H), 4.10 (dd, J=11.8, 4.6 Hz, 1H), 3.93-3.83 (m, 2H), 2.45 (br t, J=5.2 Hz, 4H), 2.31 (s, 3H), 2.21-2.09 (m, 1H), 2.00 (br d, J=14.2 Hz, 1H), 1.71-1.55 (m, 6H), 1.35 (d, J=6.8 Hz, 3H), 1.22-0.92 (m, 6H); LCMS (ESI) m/z: 577.2 (M+1).

[0265] The compounds involved in the present disclosure are IAP inhibitors. The following experimental results confirm that the compounds listed in the present patent application are IAP inhibitors and may be used as potential anticancer drugs. IC.sub.50 used herein refers to a concentration of a certain reagent, and 50% of the maximum inhibition may be achieved when the reagent is used in said concentration.

Experimental Example I

Binding Experiment of cIAP1 BIR3 and XIAP BIR3

Experimental Materials:

[0266] Buffer system for the test (a buffer for cIAP1 BIR3 or XIAP BIR3): 100 mM potassium phosphate, pH 7.5; 0.1% BSA; 0.005% Triton X-100; and 1% dimethyl sulfoxide.

[0267] Probe: ARPFAQ-K(5-FAM)-NH.sub.2.

[0268] Targets:

[0269] cIAP1-BIR3-his: RBC Cat# APT-11-370, the BIR3 domain of human cIAP1 (covering amino acids 258 to 363; cIAP1 BIR3), as a GST-fusion protein expressed and purified from E. coli.

[0270] XIAP-BIR3-his: RBC Cat# APT-11-374, the BIR3 domain of XIAP (covering amino acids 255 to 356; XIAP BIR3), as a GST-fusion protein expressed and purified from E. coli.

[0271] Reaction conditions: 5 nM ARPFAQ-K(5-FAM)-NH.sub.2, 20 nM cIAP1 BIR3 and 30 nM XIAP BIR3.

Steps of Experiment:

[0272] First, a fresh buffer for cIAP1 BIR3 or XIAP BIR3 was prepared, a cIAP1 BIR3 or XIAP BIR3 solution (2-fold diluted) was added thereto, and then a compound to be tested which had been dissolved in 100% DMSO was added into the buffer solution containing cIAP1 BIR3 or XIAP BIR3 by an acoustic technique. Thereafter, the probe (2-fold diluted) was added, and the resultant was mixed and incubated in the dark at room temperature for 60 minutes. The fluorescence polarization was measured and the mP value was calculated. Finally, the IC.sub.50 value was obtained.

Results of Experiment:

[0273] As shown in Table 1.

Conclusion of Experiment:

[0274] The compounds of the present disclosure exhibited cIAP1 BIR3-binding activity and were selective for cIAP1 and XIAP.

Experimental Example II

In-Vitro Cell Viability Test

Experimental Materials:

[0275] RPMI 1640 medium (Invitrogen-22400089); fetal bovine serum (Invitrogen-10099141); Trypsin, 0.05% (1×) with EDTA 4Na (Invitrogen-25300062); luminescent cell viability assay kit (Promega-G7573); Dulbecco's phosphate buffered saline (HyClone-SH30028.01B); and is 384-well plate (Corning-6007680). Envision Multi-label Analyzer.

Experimental Method:

[0276] 1. 30 μL of MDA-MB-231 cell suspension, which contained 250 MDA-MB-231 cells, was added into the wells of a 384-microwell plate.

[0277] 2. 20 μL of a test compound (the test compounds were formulated at a high concentration of 10 μM, the test compounds were subjected to a 5-fold gradient dilution, and each compound was diluted to 10 concentration gradients) was added, and then the cell plate was put back into a carbon dioxide incubator to be incubated for 7 days.

[0278] 3. The cell plate was kept flat at room temperature for 30 minutes.

[0279] 4. 20 μL of a Promega CellTiter-Glo reagent was added into each well of the cell plate.

[0280] 5. After 10 minutes, the Envision Multi-label Analyzer was used for data reading.

Results of Experiment: See Table 1.

Conclusion of Experiment:

[0281] The compounds of the present disclosure had anti-proliferative activity on MDA-MB-231 cells.

TABLE-US-00001 TABLE 1 Test cIAP1 BIR3 XIAP BIR3 MDA-MB-231 Compounds IC.sub.50 (nM) IC.sub.50 (nM) Cell IC.sub.50 (nM) Example 1 3.7 74.7 70.0 Example 2 5.0 29.9 54.8 Example 3 1.0 18.1 16.0 Example 4 2.6 97.0 220.0 Example 5 2.6 95.9 75.0 Example 6 58.3 9.3 497.0 Example 7 4.9 45.0 16.6 Example 8 5.6 40.3 75.0 Example 9 5.7 20.0 26.0 Example 10 4.5 27.2 32.4 Example 11 5.2 139.0 74.8 Example 13 5.0 30.6 44.3 Example 14 6.2 49.9 57.7 Example 15 4.0 258.0 93.0 Example 16 8.4 346.0 109.3 Example 17 1.9 80.7 79.0 Example 18 5.1 42.5 22.5 Example 19 4.6 129.0 43.1 Example 20 1.1 53.9 75.0 Example 21 4.2 37.4 16.4 Example 22 3.0 42.0 85.0 Example 23 4.6 21.3 45.2 Example 24 4.1 19.2 73.0 Example 25 6.8 208.0 1153.0 Example 26 1.1 60.0 181.0 Example 27 4.4 46.4 282.1 Example 28 5.2 27.3 36.7 Example 29 6.1 31.4 134.0 Example 30 4.4 21.6 47.6 Example 31 3.7 49.1 1461.0

Experimental Example III

In-Vivo Drug Efficacy Study 1

[0282] The in-vivo drug efficacy experiments were carried out in BALB/c nude mice implanted subcutaneously with human tumor cell line-derived xenograft (CDX) derived from patients suffering from MDA-MB-231 triple-negative breast cancer.

Experimental Operation:

[0283] BALB/c nude mice, female, 6 to 8 weeks old, weighing about 18 to 22 g, were kept in a special pathogen-free environment and in separated ventilated cages (3 mice per cage). All cages, bedding and water were disinfected before use. All animals had free access to standard certified commercial laboratory diets. A total of 48 mice purchased from Shanghai BK Laboratory Animal Co., LTD were used for the study. Each mouse was inoculated subcutaneously in the right flank with tumor cells (10×10.sup.6 cells in 0.2 mL of a phosphate buffer) for tumor growth. The administration was initiated when the average tumor volume reached about 147 cubic millimeters. The test compounds were orally administered daily at a dose of 30 mg/kg. Tumor volumes were measured with a two-dimensional caliper every 3 days. The volume was measured in cubic millimeters and calculated by the following formula: V=0.5 a×b.sup.2, where a and b were the long and short diameters of a tumor, respectively. Anti-tumor efficacy was determined by dividing the average increase in tumor volume of animals treated with a compound by the average increase in tumor volume of untreated animals.

Results of Experiment: See Table 2.

Conclusion of Experiment:

[0284] In the MDA-MB-231 triple-negative breast cancer CDX model used for in-vivo drug efficacy study, the compounds of the present disclosure exhibited drug efficacy.

TABLE-US-00002 TABLE 2 Dose Tumor Volume (mm.sup.3) Examples (mg/kg) Day 0 Day 6 Day 13 Day 20 Blank Control 0 148 475 1225 1750 Example 2 30 147 214 282 810

Experimental Example IV

In-Vivo Drug Efficacy Study 2

[0285] The in-vivo drug efficacy experiments were carried out in BALB/c nude mice implanted subcutaneously with human tumor cell line-derived xenograft (CDX) derived from patients suffering from MDA-MB-231 triple-negative breast cancer.

Experimental Operation:

[0286] BALB/c nude mice, female, 6 to 8 weeks old, weighing about 18 to 22 g, were kept in a special pathogen-free environment and in separated ventilated cages (3 mice per cage). All cages, bedding and water were disinfected before use. All animals had free access to standard certified commercial laboratory diets. A total of 48 mice purchased from Shanghai BK Laboratory Animal Co., LTD were used for the study. Each mouse was inoculated subcutaneously in the right flank with tumor cells (10×10.sup.6 cells in 0.2 mL of a phosphate buffer) for tumor growth. The administration was initiated when the average tumor volume reached about 110 cubic millimeters. The test compounds were orally administered daily at a dose of 30 mg/kg. Tumor volumes were measured with a two-dimensional caliper every 3 days. The volume was measured in cubic millimeters and calculated by the following formula: V=0.5 a×b.sup.2, where a and b were the long and short diameters of a tumor, respectively. Anti-tumor efficacy was determined by dividing the average increase in tumor volume of animals treated with a compound by the average increase in tumor volume of untreated animals.

Conclusion of Experiment:

[0287] In the MDA-MB-231 triple-negative breast cancer CDX model used for in-vivo drug efficacy study, the compounds of the present disclosure exhibited relatively good drug efficacy.

TABLE-US-00003 TABLE 3 Dose Tumor volume (mm.sup.3) Examples (mg/kg) Day 0 Day 5 Day 12 Day 20 Blank Control 0 110 261 526 772 Example 2 30 110 111 86 177