1H-PYRAZOLE DERIVATIVE AND APPLICATION THEREOF AS DUAL TARGET INHIBITOR OF SYK AND VEGFR2

Abstract

A class of spleen tyrosine kinase (Syk) and vascular endothelial growth factor 2 (VEGFR2) dual inhibitors, and specifically disclosed are a compound represented by formula (I), a pharmaceutically acceptable salt thereof, and an application thereof in the preparation of a Syk and VEGFR2 dual inhibitor related drug.

##STR00001##

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, ##STR00068## wherein, R.sub.1 and are each independently selected from H and pyrazolyl, and R.sub.1 and R.sub.2 are not both pyrazolyl or H; R.sub.3 and R.sub.4 are each independently selected from H, F, Cl, Br, I, OH, NH.sub.2, CN, C.sub.1-3 alkyl and C.sub.1- 3 alkoxy, wherein C.sub.1-3 alkyl and C.sub.1-3 alkoxy are optionally substituted with 1, 2 or 3 halogens; T.sub.1 is selected from CH and N; Di is selected from -O-, -C(R.sub.5)(R.sub.6)-, -N(R.sub.7)- and ##STR00069## R.sub.5 and R.sub.6 are each independently selected from H, F, Cl, Br, I, OH and C.sub.1-3 alkyl, wherein C.sub.1-3 alkyl is optionally substituted with 1, 2 or 3 halogens; alternatively, R.sub.5 and R.sub.6 together with a carbon atom to which they are both connected form an oxetanyl group; R.sub.7 is selected from H, ##STR00070## and C.sub.1-3 alkyl, wherein C.sub.1-3 alkyl is optionally substituted with 1, 2 or 3 halogens; R.sub.8 is selected from H and -C(=O)-C.sub.1-3 alkyl; and n is selected from 1 and 2.

2. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.1 is selected from H, ##STR00071## and ##STR00072## .

3. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.2 is selected from H, ##STR00073## and ##STR00074## .

4. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.3 is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN, CH.sub.3 and CH.sub.3O.

5. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.4 is selected from H, F, Cl, Br, I, OH, NH.sub.2, CN, CH.sub.3 and CH.sub.3O.

6. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.5 is selected from H, F, Cl, Br, I, OH, NH.sub.2 and CH.sub.3.

7. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.6 is selected from H, F, Cl, Br, I, OH, NH.sub.2 and CH.sub.3.

8. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.5 and R.sub.6 together with the carbon atom to which they are both connected form ##STR00075## .

9. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.7 is selected from H, CH.sub.3 and ##STR00076## .

10. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.8 is selected from H and -C(=O)-CH.sub.3.

11. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein ##STR00077## is selected from ##STR00078## ##STR00079## ##STR00080## and ##STR00081## wherein R.sub.5, R.sub.6, R.sub.7 and R.sub.8 are defined according to claim 1 .

12. The compound or the pharmaceutically acceptable salt thereof according to claim 11, wherein ##STR00082## is selected from ##STR00083## ##STR00084## ##STR00085## ##STR00086## ##STR00087## and ##STR00088## .

13. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is selected from: ##STR00089## ##STR00090## ##STR00091## and ##STR00092## wherein, R.sub.3, R.sub.4, T.sub.1, D.sub.1 and n are defined according to claim 1.

14. A compound represented by any of the following formula or a pharmaceutically acceptable salt thereof, ##STR00093## ##STR00094## ##STR00095## ##STR00096## ##STR00097## ##STR00098## ##STR00099## ##STR00100## ##STR00101## ##STR00102## ##STR00103## and ##STR00104## .

15-16. (canceled)

17. A method for preparing a compound of formula 1, comprising: ##STR00105## , step 1 of synthesizing a compound of formula 1-2 by reacting a compound of formula 1-1 with imidazole; step 2 of synthesizing a compound of formula 1-3 from the compound of formula 1-2; step 3 of synthesizing a compound of formula 1-4 from the compound of formula 1-3; step 4 of synthesizing a compound of formula 1-5 from the compound of formula 1-4; step 5 of synthesizing a compound of formula 1-6 by reacting the compound of formula 1-5 with 4-morpholinoaniline; step 6 of synthesizing a compound of formula 1-8 by reacting the compound of formula 1-6 with a compound of formula 1-7; and step 7 of synthesizing the compound of formula 1 from the compound of formula 1-8.

18. The method according to claim 17, wherein: step 1 comprises adding imidazole and DIEA to a solution of the compound of formula 1-1 in acetonitrile, stirring the reaction solution at 90° C. for 16 hours, concentrating the reaction solution, adding water, and extracting the mixture twice with ethyl acetate, and then combining, drying, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 1-2; step 2 comprises adding stannous chloride dihydrate to a solution of the compound of formula 1-2 in ethanol, stirring the reaction solution at 90° C. for 3 hours, concentrating the reaction solution under reduced pressure, adding ethyl acetate, neutralizing the solution with saturated sodium bicarbonate solution to pH=9 to precipitate a white solid, filtering off the precipitated solid, adding to the filtrate with water, extracting the filtrate twice with ethyl acetate, and combining, washing with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 1-3; step 3 comprises adding CDI to the compound of formula 1-3 in dichlorobenzene under the protection of nitrogen, stirring the reaction solution at 190° C. for 2 hours, cooling the reaction solution to room temperature, filtering the reaction solution, and drying the filter cake from the filtering to obtain the compound of formula 1-4; step 4 comprises adding the compound of formula 1-4 and dimethylaniline to phosphorus oxychloride, stirring the reaction solution at 110° C. for 1.5 hours, concentrating the reaction solution, adjusting the pH to 9 with saturated aqueous sodium bicarbonate solution to precipitate a solid, filtering the precipitated solid, and drying the filter cake from the filtering to obtain the compound of formula 1-5; step 5 comprises adding 4-morpholinoaniline and DIEA to the compound of formula 1-5 in isopropanol, stirring the reaction solution at 100° C. for 32 hours, filtering the reaction solution, and drying the filter cake from the filtering to obtain the compound of formula 1-6; step 6 comprises adding the compound of formula 1-6, the compound of formula 1-7, potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4-dioxane and water, stirring the reaction solution at 80° C. for 4 hours under the protection of nitrogen, adding the reaction solution with water, extracting the reaction solution twice with ethyl acetate, and combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 1-8; and step 7 comprises adding trifluoroacetic acid to a solution of the compound of formula 1-8 in dichloromethane, stirring the reaction solution at 25° C. for 1 hour, adjusting the pH of the reaction solution to 8 with aqueous ammonia, adding water, and extracting the reaction solution twice with dichloromethane, then combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction, adding acetone to the crude product, and heating to 60° C., stirring for 0.5 hours, cooling to room temperature, filtering, and vacuum drying the crude product to obtain the compound of formula 1.

19. A method for preparing a compound of formula 2, comprising: ##STR00106## step 1 of synthesizing a compound of formula 2-2 by reacting a compound of formula 1-6 with a compound of formula 2-1; and step 2 of synthesizing a compound of formula 2 from the compound of formula 2-2.

20. The method according to claim 19, wherein: step 1 comprises adding the compound of formula 1-6, the compound of formula 2-1, potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4-dioxane and water, stirring the reaction solution at 80° C. for 4 hours under the protection of nitrogen, adding water to the reaction solution, and extracting the mixture twice with ethyl acetate, and then combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 2-2; and step 2 comprises adding trifluoroacetic acid to a solution of the compound of formula 2-2 in dichloromethane; stirring the reaction solution at 25° C. for 2 hours, adjusting the pH of the reaction solution to 8 with aqueous ammonia, adding water, extracting the solution twice with dichloromethane, then combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction, and separating and purifying the crude product by high performance liquid chromatography to obtain the compound of formula 2.

21. A method for preparing a compound of formula 3, comprising: ##STR00107## ,step 1 of synthesizing a compound of formula 3-2 by reacting a compound of formula 3-1 with imidazole; step 2 of synthesizing a compound of formula 3-3 from the compound of formula 3-2; step 3 of synthesizing a compound of formula 3-4 from the compound of formula 3-3; step 4 of synthesizing a compound of formula 3-5 from the compound of formula 3-4; step 5 of synthesizing a compound of formula 3-6 by reacting the compound of formula 3-5 with 4-morpholinoaniline; and step 6 of synthesizing the compound of formula 3 by reacting the compound of formula 3-6 with a compound of formula 1-7.

22. The method according to claim 21, wherein: step 1 comprises adding imidazole and DIEA to a solution of the compound of formula 3-1 in acetonitrile, stirring the reaction solution at 90° C. for 16 hours, concentrating the reaction solution, adding water, extracting the mixture twice with ethyl acetate, and then combining, washing once with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 3-2; step 2 comprises adding stannous chloride dihydrate to a solution of the compound of formula 3-2 in ethanol, stirring the reaction solution at 80° C. for 3 hours, concentrating the reaction solution under reduced pressure, adding ethyl acetate, neutralizing the solution with saturated sodium bicarbonate solution to pH=9 to precipitate a white solid, filtering off the precipitated solid, adding water to the filtrate, extracting the filtrate twice with ethyl acetate, and combining, washing with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 3-3; step 3 comprises adding CDI to the compound of formula 3-3 in dichlorobenzene under the protection of nitrogen, stirring the reaction solution at 190° C. for 2 hours, cooling the reaction solution to room temperature, filtering the reaction solution, and drying the filter cake from the filtering to obtain the compound of formula 3-4; step 4 comprises adding the compound of formula 3-4 and dimethylaniline to phosphorus oxychloride, stirring the reaction solution at 110° C. for 1.5 hours, concentrating the reaction solution, adjusting the pH to 9 with saturated aqueous sodium bicarbonate solution to precipitate a solid, filtering the precipitated solid, drying the filter cake from the filtering, and purifying the filter cake by column chromatography to obtain the compound of formula 3-5; step 5 comprises adding 4-morpholinoaniline and DIEA to the compound of formula 3-5 in isopropanol, stirring the reaction solution at 100° C. for 16 hours, filtering the reaction solution, and drying the filter cake from the filtering to obtain the compound of formula 3-6; and step 6 comprises adding the compound of formula 3-6, the compound of formula 1-7, potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4-dioxane and water, stirring the reaction solution at 80° C. under the protection of nitrogen, adding water to the reaction solution, extracting the reaction solution twice with ethyl acetate, then combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and separating and purifying the crude product by high performance liquid chromatography to obtain the compound of formula 3.

23. A method for preparing a compound of formula 4, comprising synthesizing the compound of formula 4 by reacting a compound of formula 3-6 with a compound of formula 2-1, ##STR00108## .

24. The method according to claim 23, comprising adding the compound of formula 3–6, the compound of formula 2–1, potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4–dioxane and water, stirring the reaction solution at 90° C. for 4 hours under the protection of nitrogen, adding water to the reaction solution, and extracting the mixture twice with ethyl acetate, then combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and seperating and purifying the crude product by high performance liquid chromatography to obtain the compound of formula 4.

25. Amethod for preparing a compound of formula 5, comprising: ##STR00109## , step 1 of synthesizing a compound of formula 5-3 by reacting a compound of formula 5-1 with a compound of formula 5-2; step 2 of synthesizing a compound of formula 5-4 from the compound of formula 5-3; step 3 of synthesizing a compound of formula 5-5 from the compound of formula 5-4; step 4 of synthesizing a compound of formula 5-6 from the compound of formula 5-5; step 5 of synthesizing a compound of formula 5-7 by reacting the compound of formula 5-5 with the compound of formula 5-6; step 6 of synthesizing a compound of formula 5-9 by reacting the compound of formula 5-7 with a compound of formula 5-8; and step 7 of synthesizing the compound of formula 5 from the compound of formula 5-9.

26. The method according to claim 25, wherein: step 1 comprises adding potassium carbonate and the compound of formula 5-2 to a solution of the compound of formula 5-1 in DMSO, stirring the reaction solution at 80° C. for 16 hours, concentrating the reaction solution, adding water, and extracting the mixture three times with ethyl acetate, and then combining, drying, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 5-3; step 2 comprises adding the compound of formula 5-3 in small batches to a dioxane hydrochloride solution, stirring the mixture at room temperature for 16 hours, and concentrating the reaction solution under reduced pressure to obtain the compound of formula 5-4; step 3 comprises adding sodium acetate to a solution of the compound of formula 5-4 in ethanol under the protection of nitrogen, stirring the mixture at room temperature for 1 hour before adding oxetanone and zinc chloride, then stirring the mixture at room temperature for 2 hours before adding sodium cyanoborohydride, stirring the mixture at 40° C. for 16 hours, then adding water to the reaction solution, extracting the mixture three times with ethyl acetate, and then combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 5-5; step 4 comprises adding palladium carbon to a solution of the compound of formula 5-5 in methanol under the protection of nitrogen, then replacing the nitrogen with hydrogen by a hydrogen balloon three times, stirring the reaction at room temperature for 16 hours under hydrogen, then filtering the reaction solution, and concentrating and drying the filtrate to obtain the compound of formula 5-6; step 5 comprises adding the compound of formula 5-6 and DIEA to a solution of the compound of formula 1-5 in isopropanol, stirring the reaction solution at 100° C. for 16 hours, cooling the reaction solution to room temperature, filtering the reaction solution, and drying the filter cake from the filtering to obtain the compound of formula 5-7; step 6 comprises adding the compound of formula 5-7, the compound of formula 5-8, potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4-dioxane and water, stirring the reaction solution at 100° C. for 16 hours under the protection of nitrogen, adding water to the reaction solution, extracting the mixture three times with ethyl acetate, and combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 5-9; and step 7 comprises adding tetrabutylammonium fluoride and ethylenediamine to a solution of the compound of formula 5-9 in tetrahydrofuran, stirring the reaction solution at 75° C. for 16 hours, adjusting the pH of the reaction solution to 8 with sodium hydroxide solution, adding water, extracting the solution twice with dichloromethane, then combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and separating the crude product by silica gel column chromatography to obtain the compound of formula 5.

27. A method for preparing a compound of formula 6, comprising: ##STR00110## step 1 of synthesizing a compound of formula 6-2 by reacting a compound of formula 6-1 with a compound of formula 1-5; step 2 of synthesizing a compound of formula 6-3 by reacting the compound of formula 6-2 with a compound of formula 5-8; and step 3 of synthesizing the compound of formula 6 from the compound of formula 6-3.

28. The method according to claim 27, wherein: step 1 comprises adding the compound of formula 6-1 and DIEA to a solution of compound of formula 1-5 in dimethyl sulfoxide, stirring the reaction solution at 120° C. for 16 hours, cooling the reaction solution to room temperature, then adding water to precipitate a solid, filtering the mixture, slurrying the filter cake from the filtering with isopropanol, and filtering and drying the slurry to obtain the compound of formula 6-2; step 2 comprises adding the compound of formula 6-2, the compound of formula 5-8, potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4-dioxane and water, then stirring the reaction solution at 100° C. for 16 hours under the protection of nitrogen, adding water to the reaction solution, extracting the solution three times with ethyl acetate, combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and separating the crude product by silica gel column chromatography to obtain the compound of formula 6-3; and step 3 comprises adding the compound of formula 6-3 to dioxane hydrochloride, stirring the reaction solution at 25° C. for 16 hours, adjusting the pH of the reaction solution to 8 with aqueous sodium hydroxide solution, adding water, extracting the solution twice with dichloromethane, then combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, adding acetone to the crude product, stirring the mixture for 0.5 hours, filtering the mixture, and drying the filter cake from the filtering to obtain the compound of formula 6.

29. A method for preparing a compound of formula 7, comprising: ##STR00111## step 1 of synthesizing a compound of formula 7-2 by reacting a compound of formula 7-1 with a compound of formula 1-5; step 2 of synthesizing a compound of formula 7-3 by reacting the compound of formula 7-2 with a compound of formula 5-8; and step 3 of synthesizing the compound of formula 7 from the compound of formula 7-3.

30. The method according to claim 29, wherein: step 1 comprises adding the compound of formula 7-1 and DIEA to a solution of compound of formula 1-5 in dimethyl sulfoxide, stirring the reaction solution at 120° C. for 16 hours, cooling the reaction solution to room temperature, then adding water to precipitate a solid, filtering the mixture, slurrying the filter cake from the filtering with isopropanol, and filtering and drying the slurry to obtain the compound of formula 7-2; step 2 comprises adding the compound of formula 7-2, the compound of formula 5-8, potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4-dioxane and water, then stirring the reaction solution at 100° C. for 16 hours under the protection of nitrogen, adding water to the reaction solution, extracting the solution three times with ethyl acetate, combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and separating the crude product by silica gel column chromatography to obtain the compound of formula 7-3; and step 3 comprises adding the compound of formula 7-3 to dioxane hydrochloride, stirring the reaction solution at 25° C. for 16 hours, adjusting the pH of the reaction solution to 8 with aqueous sodium hydroxide solution, adding water, extracting the solution twice with dichloromethane, then combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and separating the crude product by silica gel column chromatography to obtain the compound of formula 7.

31. A method for preparing a compound of formula 8, comprising: ##STR00112## step 1 of synthesizing a compound of formula 8-2 by reacting a compound of formula 8-1 with a compound of formula 5-1; step 2 of synthesizing a compound of formula 8-3 from the compound of formula 8-2; step 3 of synthesizing a compound of formula 8-4 by reacting the compound of formula 8-3 with a compound of formula 1-5; step 4 of synthesizing a compound of formula 8-5 by reacting the compound of formula 8-4 with a compound of formula 5-8; and step 5 of synthesizing the compound of formula 8 from the compound of formula 8-5.

32. The method according to claim 31, wherein: step 1 comprises adding potassium carbonate and the compound of formula 8-1 to a solution of the compound of formula 5-1 in DMSO, stirring the reaction solution at 100° C. for 16 hours, concentrating the reaction solution, adding water, and extracting the mixture three times with ethyl acetate, and combining, drying, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 8-2; step 2 comprises adding ammonium chloride and iron powder to a solution of the compound of formula 8-2 in ethanol and water under the protection of nitrogen, stirring the reaction solution at 100° C. for 16 hours, then filtering the reaction solution, adding water to the filtrate, extracting the mixture three times with ethyl acetate, and combining, drying, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 8-3; step 3 comprises adding the compound of formula 8-3 and DIEA to a solution of the compound of formula 1-5 in dimethyl sulfoxide, stirring the reaction solution at 120° C. for 16 hours, then cooling the reaction solution to room temperature, adding water to precipitate a solid, filtering the mixture, slurrying the filter cake from the filtering with isopropanol, and filtering and drying the slurry to obtain the compound of formula 8-4; step 4 comprises adding the compound of formula 8-4, the compound of formula 5-8, potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4-dioxane and water, then stirring the reaction solution at 100° C. for 16 hours under the protection of nitrogen, adding water to the reaction solution, extracting the solution three times with dichloromethane, combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and separating the crude product by silica gel column chromatography to obtain the compound of formula 8-5; and step 5 comprises adding the compound of formula 8-5 to dioxane hydrochloride, stirring the reaction solution at 25° C. for 16 hours, then adjusting the pH of the reaction solution to 8 with aqueous sodium hydroxide solution, adding water, extracting the solution twice with dichloromethane, then combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and separating the crude product by silica gel column chromatography to obtain the compound of formula 8.

33. A method for preparing a compound of formula 9, comprising: ##STR00113## step 1 of synthesizing a compound of formula 9-2 from a compound of formula 9-1; step 2 of synthesizing a compound of formula 9-3 from the compound of formula 9-2; step 3 of synthesizing a compound of formula 9-4 from the compound of formula 9-3; step 4 of synthesizing a compound of formula 9-5 by reacting the compound of formula 9-4 with a compound of formula 5-1; step 5 of synthesizing a compound of formula 9-6 from the compound of formula 9-5; step 6 of synthesizing a compound of formula 9-7 by reacting the compound of formula 9-6 with a compound of formula 1-5; and step 7 of synthesizing the compound of formula 9 by reacting the compound of formula 9-7 with a compound of formula 9-8.

34. The method according to claim 33, wherein: step 1 comprises cooling a solution of the compound of formula 9-1 in methanol and water to 0° C., adding sodium periodate to the solution, stirring the reaction solution at 20° C. for 16 hours, filtering the reaction solution, washing the filter cake from the filtering three times with ethyl acetate, and combining, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the washing to obtain the compound of formula 9-2; step 2 comprises adding trifluoroacetamide, rhodium acetate, magnesium oxide diacetoxyiodobenzene and potassium carbonate to a solution of the compound of formula 9-2 in dichloromethane under the protection of nitrogen, stirring the reaction solution at 20° C. for 16 hours, then filtering the reaction solution, washing the filter cake from the filtering three times with dichloromethane, combining, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the washing to obtain a crude product, and separating and purifying the crude product by silica gel column chromatography to obtain the compound of formula 9-3; step 3 comprises adding the compound of formula 9-3 to a solution of hydrobromic acid in acetic acid, stirring the reaction solution at 20° C. for 10 hours, then adjusting the pH of the reaction solution to 8 with aqueous sodium hydroxide solution, adding water, extracting the solution three times with dichloromethane, and combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 9-4; step 4 comprises adding potassium carbonate and the compound of formula 9-4 to a solution of the compound of formula 5-1 in DMSO, stirring the reaction solution at 100° C. for 16 hours, concentrating the reaction solution, adding water, and extracting the mixture three times with ethyl acetate, and combining, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 9-5, step 5 comprises adding ammonium chloride and iron powder to a solution of the compound of formula 9-5 in ethanol and water under the protection of nitrogen, stirring the reaction solution at 90° C.1 hour, then filtering the reaction solution, adding water to the filtrate, extracting the mixture three times with ethyl acetate, and combining, drying, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 9-6; step 6 comprises adding the compound of formula 9-6 and DIEA to a solution of the compound of formula 1-5 in dimethyl sulfoxide, stirring the reaction solution at 120° C. for 16 hours, then cooling the reaction solution to room temperature, adding water to precipitate a solid, filtering the mixture, slurrying the filter cake from the filtering with isopropanol, and filtering and drying the slurry to obtain the compound of formula 9-7; and step 7 comprises adding the compound of formula 9-7, the compound of formula 9-8 potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4-dioxane and water, then stirring the reaction solution at 100° C. for 16 hours under the protection of nitrogen, adding water to the reaction solution, extracting the solution three times with dichloromethane, combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and separating and purifying the crude product by silica gel column chromatography to obtain the compound of formula 9.

35. A method for preparing a compound of formula 10, comprising: ##STR00114## step 1 of synthesizing a compound of formula 10-1 from a compound of formula 9-5; step 2 of synthesizing a compound of formula 10-2 from the compound of formula 10-1; step 3 of synthesizing a compound of formula 10-3 by reacting the compound of formula 10-2 with a compound of formula 1-5; and step 4 of synthesizing the compound of formula 10 by reacting the compound of formula 10-3 with a compound of formula 9-8.

36. The method according to claim 35, wherein: step 1 comprises adding triethylamine and acetyl chloride to a solution of the compound of formula 9-5 in dichloromethane, stirring the reaction solution at 25° C. for 2 hours, adding water to the reaction solution, extracting the mixture three times with dichloromethane, and combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 10-1; step 2 comprises adding ammonium chloride and iron powder to a solution of the compound of formula 10-1 in ethanol and water under the protection of nitrogen, stirring the reaction solution at 90° C.1 hour, then filtering the reaction solution, adding water to the filtrate, extracting the mixture three times with ethyl acetate, and combining, drying, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 10-2; step 3 comprises adding the compound of formula 10-2 and DIEA to a solution of the compound of formula 1-5 in dimethyl sulfoxide, stirring the reaction solution at 120° C. for 16 hours, then cooling the reaction solution to room temperature, adding water to precipitate a solid, filtering the mixture, slurrying the filter cake from the filtering with isopropanol, and filtering and drying the slurry to obtain the compound of formula 10-3; and step 4 comprises adding the compound of formula 10-3, the compound of formula 9-8, potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4-dioxane and water, then stirring the reaction solution at 100° C. for 16 hours under the protection of nitrogen, adding water to the reaction solution, extracting the solution three times with dichloromethane, combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and separating and purifying the crude product by silica gel column chromatography to obtain the compound of formula 10.

37. A method for preparing a compound of formula 11, comprising: ##STR00115## step 1 of synthesizing a compound of formula 11-2 by reacting the compound of formula 11-1 with a compound of formula 1-5; and step 2 of synthesizing the compound of formula 11 by reacting the compound of formula 11-2 with a compound of formula 9-8.

38. The method according to claim 37, wherein: step 1 comprises mixing the compound of formula 1-5 and the compound of formula 11-1 and stirring the mixture at 120° C. for 16 hours under the protection of nitrogen, cooling the reaction solution to room temperature, adding dichloromethane and methanol and stirring the mixture for 15 minutes, filtering the mixture, collecting and drying the filter cake from the filtering under vacuum to obtain the compound of formula 11-2; and step 2 comprises adding the compound of formula 11-2, the compound of formula 9-8, potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4-dioxane and water, then stirring the reaction solution at 100° C. for 16 hours under the protection of nitrogen, adding water to the reaction solution, extracting the solution three times with dichloromethane, combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and separating and purifying the crude product by silica gel column chromatography to obtain the compound of formula 11.

39. A method for preparing a compound of formula 12, comprising: ##STR00116## step 1 of synthesizing a compound of formula 12-2 by reacting a compound of formula 12-1 with a compound of formula 5-1; step 2 of synthesizing a compound of formula 12-3 from the compound of formula 12-2; step 3 of synthesizing a compound of formula 12-4 by reacting the compound of formula 12-3 with a compound of formula 1-5; step 4 of synthesizing a compound of formula 12-5 by reacting the compound of formula 12-4 with a compound of formula 5-8; and step 5 of synthesizing the compound of formula 12 from the compound of formula 12-5.

40. The method according to claim 39, wherein: step 1 comprises adding potassium carbonate and the compound of formula 12-1 to a solution of the compound of formula 5-1 in DMSO, stirring the reaction solution at 100° C. for 16 hours, concentrating the reaction solution, adding water, and extracting the mixture three times with ethyl acetate, and combining, drying, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 12-2; step 2 comprises adding ammonium chloride and iron powder to a solution of the compound of formula 12-2 in ethanol and water under the protection of nitrogen, stirring the reaction solution at 100° C. for 16 hours, then filtering the reaction solution, adding water to the filtrate, extracting the mixture three times with ethyl acetate, and combining, drying, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 12-3; step 3 comprises adding the compound of formula 12-3 and DIEA to a solution of the compound of formula 1-5 in isopropanol, stirring the reaction solution at 120° C. for 16 hours, then cooling the reaction solution to room temperature, adding water, extracting the solution three times with ethyl acetate, and combining, drying, filtering and concentrating the organic phases from the extraction to obtain the compound of formula 12-4; step 4 comprises adding the compound of formula 12-4, the compound of formula 5-8, potassium carbonate and Pd(dppf)Cl.sub.2 to 1,4-dioxane and water, then stirring the reaction solution at 100° C. for 16 hours under the protection of nitrogen, adding water to the reaction solution, extracting the solution three times with dichloromethane, combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases from the extraction to obtain a crude product, and separating the crude product by silica gel column chromatography to obtain the compound of formula 12-5; and step 5 comprises adding the compound of formula 12-5 to dioxane hydrochloride, stirring the reaction solution at 25° C. for 16 hours, then adjusting the pH of the reaction solution to 8 with aqueous sodium hydroxide solution, adding water, extracting the solution twice with dichloromethane, and then combining, washing twice with saturated brine, drying over anhydrous sodium sulfate, filtering and concentrating the organic phases to obtain the compound of formula 12.

41. Use of the compound or pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a spleen tyrosine kinase (Syk) and vascular endothelial growth factor 2 dual inhibitor related pharmaceutical.

42. The use according to claim 41, wherein the spleen tyrosine kinase and vascular endothelial growth factor 2 dual inhibitor related pharmaceutical is used for the treatment of allergic, autoimmune and inflammatory diseases such as dry eye and allergic conjunctivitis, retinal inflammatory diseases, age-related macular degeneration, proliferative diabetic retinopathy and retinopathy of prematurity, cancer, rheumatoid arthritis, glomerulonephritis, multiple vasculitides, idiopathic thrombocytopenic purpura, myasthenia gravis, allergic rhinitis, chronic obstructive pulmonary disease, adult respiratory distress syndrome and asthma.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0074] FIG. 1 shows the amounts of tear secretion on the 7th and 12th days after modeling. Note: PRE (D-1) means the day before modeling, D7 means the 7th day, D12 means the 12th day; p means significant difference.

[0075] FIG. 2 shows the corneal fluorescence staining scores on the 7th and 12th days after modeling. Note: PRE (D-1) means the day before modeling, D7 means the 7th day, D12 means the 12th day; p means significant difference.

DETAILED DESCRIPTION

[0076] The invention will be described in detail by the following examples which do not adversely limit the invention in any way. The invention has been described in detail herein, and specific embodiments thereof have also been disclosed. Various changes and modifications, which are made to the specific embodiments of the invention without departing from the spirit and scope of the invention, are apparent to those skilled in the art.

Example 1: Preparation of Compound 1

[0077] ##STR00056##

Step 1: Synthesis of Compound 1-2

[0078] Imidazole (1.55 g, 22.73 mmol), and DIEA (2.94 g, 22.73 mmol) were added to a solution of 4-bromo-2-fluoro-1-nitrobenzene (5 g, 22.73 mmol) in acetonitrile (100 mL). The reaction solution was stirred at 90° C. for 16 hours. The reaction solution was concentrated, water (100 mL) was added, and the mixture was extracted twice with ethyl acetate (50 mL). The organic phases were combined, dried, filtered and concentrated to obtain compound 1-2.

[0079] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 7.79 (s, 1H), 7.34 (s, 1H), 7.30 (dd, J=2.4, 8.7 Hz, 1H), 7.24 (d, J=2.3 Hz, 1H), 7.11 (s, 1H), 6.83 (d, J=8.5 Hz, 1H), 5.18 (s, 2H).

Step 2: Synthesis of Compound 1-3

[0080] Stannous chloride dihydrate (20.2 g, 89.52 mmol) was added to a solution of compound 1-2 (4.00 g, 14.92 mmol) in ethanol (240 mL). The reaction solution was stirred at 90° C. for 3 hours. The reaction solution was concentrated under reduced pressure, ethyl acetate (300 mL) was added, the solution was neutralized with saturated sodium bicarbonate solution to pH=9, and a white solid was precipitated. The precipitated solid was filtered off. The filtrate was added with water (100 mL) and extracted twice with ethyl acetate (100 mL). The organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 1-3.

[0081] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 7.79 (s, 1H), 7.34 (s, 1H), 7.30 (dd, J=2.4, 8.7 Hz, 1H), 7.24 (d, J=2.3 Hz, 1H), 7.11 (s, 1H), 6.83 (d, J=8.5 Hz, 1H), 5.18 (s, 2H).

Step 3: Synthesis of Compound 1-4

[0082] Under the protection of nitrogen, CDI (749.18 mg, 4.62 mmol) was added to compound 1-3 (1.1 g, 4.62 mmol) in dichlorobenzene (20 mL). The reaction solution was stirred at 190° C. for 2 hours. The reaction solution was cooled to room temperature, filtered, and the filter cake was dried to obtain compound 1-4.

[0083] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 11.95 (s, 1H), 8.61 (d, J=1.0 Hz, 1H), 8.45 (d, J=2.0 Hz, 1H), 7.62 - 7.54 (m, 2H), 7.34 - 7.25 (m, 1H).

Step 4: Synthesis of Compound 1-5

[0084] Compound 1-4 (300 mg, 1.14 mmol) and dimethylaniline (621.65 mg, 5.13 mmol) were added to phosphorus oxychloride (5 mL). The reaction solution was stirred at 110° C. for 1.5 hours. The reaction solution was concentrated, and the pH was adjusted to 9 with saturated aqueous sodium bicarbonate solution, and a solid was precipitated, the precipitated solid was filtered, and the filter cake was dried to obtain compound 1-5.

[0085] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 9.01 (s, 1H), 8.80 (d, J=2.0 Hz, 1H), 7.95 (d, J=8.8 Hz, 1H), 7.91 (s, 1H), 7.84 (dd, J=2.0, 8.8 Hz, 1H).

Step 5: Synthesis of Compound 1-6

[0086] 4-morpholinoaniline (170.33 mg, 955.68 .Math.mol) and DIEA (148.21 mg, 1.15 mmol) were added to compound 1-5 (270.0 mg, 955.68 .Math.mol) in isopropanol (3 mL). The reaction solution was stirred at 100° C. for 32 hours. The reaction solution was filtered, and the filter cake was dried to obtain compound 1-6.

[0087] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 9.65 (s, 1H), 8.74 (d, J=1.0 Hz, 1H), 8.48 (s, 1H), 7.99 (d, J=9.0 Hz, 2H), 7.59 - 7.55 (m, 2H), 7.08 - 6.91 (m, 3H), 3.78 - 3.72 (m, 4H), 3.10-3.05 (m, 4H).

Step 6: Synthesis of Compound 1-8

[0088] Compound 1-6 (100 mg, 235.69 .Math.mol), compound 1-7 (83.19 mg, 282.83 .Math.mol), potassium carbonate (97.72 mg, 707.07 .Math.mol) and Pd(dppf)Cl.sub.2 (17.25 mg, 23.57 .Math.mol) were added to 1,4-dioxane (4 mL) and water (1 mL). Under the protection of nitrogen, the reaction solution was stirred at 80° C. for 4 hours. The reaction solution was added with water (15 mL), and extracted twice with ethyl acetate (15 mL). The organic phases were combined, washed twice with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 1-8.

Step 7: Synthesis of Compound 1

[0089] Trifluoroacetic acid (1.54 g, 13.51 mmol) was added to a solution of compound 1-8 (110.0 mg, 215.02 .Math.mol) in dichloromethane (1 mL). The reaction solution was stirred at 25° C. for 1 hour. The pH of the reaction solution was adjusted to 8 with aqueous ammonia, water (10 mL) was added, and the solution was extracted twice with dichloromethane (10 mL). The organic phases were combined and washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was added with 3 mL of acetone, heated to 60° C., stirred for 0.5 hours, then cooled to room temperature, filtered, and vacuum dried to obtain compound 1.

[0090] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ: 8.81 (s, 1H), 8.43 (s, 1H), 8.23 (s, 2H), 7.92 (d, J=8.8 Hz, 2H), 7.80 (s, 1H), 7.77 - 7.72 (m, 1H), 7.71 - 7.63 (m, 1H), 7.07 (d, J=8.8 Hz, 2H), 3.79 - 3.76 (m, 4H), 3.16 (br. s., 4H); MS (ESI) m/z: 412 [M+H].sup.+.

Example 2: Preparation of Compound 2

[0091] ##STR00057##

Step 1: Synthesis of Compound 2-2

[0092] Compound 1-6 (140 mg, 329.96 .Math.mol), compound 2-1 (116.47 mg, 395.95 .Math.mol), potassium carbonate (45.60 mg, 329.96 .Math.mol) and Pd(dppf)Cl.sub.2 (24.14 mg, 33 .Math.mol) were added to 1,4-dioxane (4 mL) and water (1 mL). Under the protection of nitrogen, the reaction solution was stirred at 80° C. for 4 hours. The reaction solution was added with water (15 mL), and extracted twice with ethyl acetate (15 mL). The organic phases were combined, washed twice with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 2-2. MS (ESI) m/z:512 [M+H].sup.+.

Step 2: Synthesis of Compound 2

[0093] Trifluoroacetic acid (23.10 g, 202.57 mmol) was added to a solution of compound 2-2 (150.0 mg, 293.22 .Math.mol) in dichloromethane (1 mL). The reaction solution was stirred at 25° C. for 2 hours. The pH of the reaction solution was adjusted to 8 with aqueous ammonia, water (10 mL) was added, and the solution was extracted twice with dichloromethane (20 mL). The organic phases were combined and washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated and purified by high performance liquid chromatography (column: 3-Phenomenex Luna C18 75x30 mm×3 .Math.m; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; B (acetonitrile)%: 45%- 75%, 7 min) to obtain compound 2 (trifluoroacetate).

[0094] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ: 9.76 (br. s., 1H), 8.81 (s, 1H), 8.54 (s, 1H), 8.06 - 7.90 (m, 3H), 7.81 - 7.63 (m, 3H), 7.01 (d, J=8.3 Hz, 2H), 6.92 (s, 1H), 3.77 (br. s., 4H), 3.12 (br. s., 2H); MS (ESI) m/z: 412 [M+H].sup.+.

Example 3: Preparation of Compound 3

[0095] ##STR00058##

Step 1: Synthesis of Compound 3-2

[0096] Imidazole (3.09 g, 45.45 mmol), and DIEA (5.87 g, 45.45 mmol)) were added to a solution of 4-bromo-1-fluoro-2-nitrobenzene (10 g, 45.45 mmol) in acetonitrile (200 mL). The reaction solution was stirred at 90° C. for 16 hours. The reaction solution was concentrated, water (100 mL) was added, and the mixture was extracted twice with ethyl acetate (50 mL). The organic phases were combined, washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 3-2.

[0097] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 8.19 - 8.09 (m, 1H), 7.86 (dd, J=2.3, 8.5 Hz, 1H), 7.67 - 7.56 (m, 1H), 7.39 - 7.34 (m, 1H), 7.23 (s, 1H), 7.05 (t, J=1.3 Hz, 1H).

Step 2: Synthesis of Compound 3-3

[0098] Stannous chloride dihydrate (36.87 g, 163.38 mmol) was added to a solution of compound 3-2 (7.30 g, 27.23 mmol) in ethanol (350 mL). The reaction solution was stirred at 80° C. for 3 hours. The reaction solution was concentrated under reduced pressure, ethyl acetate (300 mL) was added, the solution was neutralized with saturated sodium bicarbonate solution to pH=9, and a white solid was precipitated. The precipitated solid was filtered off. The filtrate was added with water (100 mL) and extracted twice with ethyl acetate (100 mL). The organic phases were combined, washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 3-3.

Step 3: Synthesis of Compound 3-4

[0099] Under the protection of nitrogen, CDI (2.23 g, 13.78 mmol) was added to compound 3-3 (1.64 g, 6.89 mmol) in dichlorobenzene (40 mL). The reaction solution was stirred at 190° C. for 2 hours. The reaction solution was cooled to room temperature, filtered, and the filter cake was dried to obtain compound 3-4.

[0100] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 8.54 (d, J=1.0 Hz, 1H), 8.08 (d, J=8.5 Hz, 1H), 7.60 (d, J=1.3 Hz, 1H), 7.52 (d, J=2.0 Hz, 1H), 7.47 (dd, J=2.1, 8.7 Hz, 1H), 7.06 (s, 1H).

Step 4: Synthesis of Compound 3-5

[0101] Compound 3-4 (800 mg, 3.03 mmol) and dimethylaniline (1.66 g, 13.70 mmol) were added to phosphorus oxychloride (10 mL). The reaction solution was stirred at 110° C. for 1.5 hours. The reaction solution was concentrated, and the pH was adjusted to 9 with saturated aqueous sodium bicarbonate solution, and a solid was precipitated, the precipitated solid was filtered, and the filter cake was dried and purified by column chromatography (silica gel, petroleum ether/ethyl acetate=100/1 to 2:1) to obtain compound 3-5.

[0102] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 8.98 (s, 1H), 8.41 (d, J=8.8 Hz, 1H), 8.25 (d, J=1.8 Hz, 1H), 7.99 (dd, J=2.0, 8.8 Hz, 1H), 7.92 (s, 1H).

Step 5: Synthesis of Compound 3-6

[0103] 4-morpholinoaniline (174.12 mg, 976.92 .Math.mol) and DIEA (126.26 mg, 976.92 .Math.mol) were added to compound 3-5 (230.0 mg, 814.10 .Math.mol) in isopropanol (5 mL). The reaction solution was stirred at 100° C. for 16 hours. The reaction solution was filtered, and the filter cake was dried to obtain compound 3-6.

[0104] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 9.72 (s, 1H), 8.68 (d, J=1.3 Hz, 1H), 8.13 (d, J=8.8 Hz, 1H), 8.00 (d, J=9.0 Hz, 2H), 7.80 (d, J=2.3 Hz, 1H), 7.71 (d, J=1.3 Hz, 1H), 7.52 (dd, J=2.1, 8.7 Hz, 1H), 6.95 (d, J=9.0 Hz, 2H), 3.79 - 3.70 (m, 4H), 3.12 - 3.00 (m, 4H).

Step 6: Synthesis of Compound 3

[0105] Compound 3-6 (90 mg, 212.12 .Math.mol), compound 1-7 (74.87 mg, 254.54 .Math.mol), potassium carbonate (87.95 mg, 636.36 .Math.mol) and Pd(dppf)Cl.sub.2 (15.52 mg, 21.21 .Math.mol) were added to 1,4-dioxane (4 mL) and water (1 mL). Under the protection of nitrogen, the reaction solution was stirred at 80° C. for 4 hours. The reaction solution was added with water (15 mL), and extracted twice with ethyl acetate (15 mL). The organic phases were combined and washed twice with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated and purified by high performance liquid chromatography (column: 3-Phenomenex Luna C18 75x30 mmx3 .Math.m ; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; B (acetonitrile)%: 45%- 75%, 7 min) to obtain compound 3 (trifluoroacetate).

[0106] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ: 8.74 (s, 1H), 8.25 - 8.12 (m, 3H), 8.01 - 7.87 (m, 3H), 7.77 (s, 1H), 7.70 (d, J=8.3 Hz, 1H), 7.08 (d, J=8.3 Hz, 2H), 3.79 (d, J=4.3 Hz, 4H), 3.17 (br. s., 4H); MS (ESI) m/z: 412 [M+H].sup.+.

Example 4: Preparation of Compound 4

[0107] ##STR00059##

[0108] Compound 3-6 (100 mg, 235.69 .Math.mol), compound 2-1 (83.19 mg, 282.83 .Math.mol), potassium carbonate (97.72 mg, 707.07 .Math.mol) and Pd(dppf)Cl.sub.2 (17.25 mg, 23.57 .Math.mol) were added to 1,4-dioxane (4 mL) and water (2 mL). Under the protection of nitrogen, the reaction solution was stirred at 90° C. for 4 hours. The reaction solution was added with water (20 mL), and extracted twice with ethyl acetate (20 mL). The organic phases were combined and washed twice with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated and purified by high performance liquid chromatography (column: 3-Phenomenex Luna C18 75×30 mm×3 .Math.m ; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; B (acetonitrile)%: 45%- 75%, 7 min) to obtain compound 4 (trifluoroacetate).

[0109] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ: 8.76 (s, 1H), 8.24 (d, J=8.5 Hz, 1H), 8.17 (s, 1H), 7.95 (d, J=8.0 Hz, 2H), 7.88 (dd, J=1.6, 8.4 Hz, 1H), 7.82 - 7.73 (m, 2H), 7.10 (d, J=8.8 Hz, 2H), 6.85 (d, J=2.3 Hz, 1H), 3.79 (br. s., 4H), 3.22 - 3.13 (m, 4H); MS (ESI) m/z: 412 [M+H].sup.+.

Example 5: Preparation of Compound 5

[0110] ##STR00060##

Step 1: Synthesis of Compound 5-3

[0111] Potassium carbonate (5.88 g, 42.52 mmol), and tert-butyl piperazine-1-carboxylate (3.96 g, 21.26 mmol) were added to a solution of p-fluoronitrobenzene (2 g, 14.17 mmol) in DMSO (20 mL). The reaction solution was stirred at 80° C. for 16 hours. The reaction solution was concentrated, water (100 mL) was added, and the mixture was extracted three times with ethyl acetate (50 mL). The organic phases were combined, dried, filtered and concentrated to obtain compound 5-3.

[0112] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=8.06 (d, J=8.8 Hz, 2H), 7.01 (d, J=9.6 Hz, 2H), 3.47 (s, 8H), 1.42 (s, 9H).

Step 2: Synthesis of Compound 5-4

[0113] Compound 5-3 (4.2 g, 13.67 mmol) was added in small batches to a dioxane hydrochloride solution (4 M, 59.81 mL), and the mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure to obtain compound 5-4 (crude hydrochloride), which was directly used in the next reaction.

[0114] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ = 9.45 (br, 1H), 8.11 (d, J=9.6 Hz, 2H), 7.10 (d, J=9.6 Hz, 2H), 3.67-3.69 (m, 4H), 3.20-3.22 (m, 4H)).

Step 3: Synthesis of Compound 5-5

[0115] Under the protection of nitrogen , sodium acetate (4.04 g, 49.22 mmol) was added to a solution of compound 5-4 (3.4 g, hydrochloride) in ethanol (40 mL), and the mixture was stirred at room temperature for 1 hour before oxetanone (1.77 g, 14.61 mmol) and zinc chloride (4.47 g, 32.81 mmol) were added, and the mixture was then stirred at room temperature for 2 hours before sodium cyanoborohydride (3.09 g, 49.22 mmol) was added. The reaction solution was stirred at 40° C. for 16 hours. The reaction solution was added with water (150 mL), and extracted three times with ethyl acetate (100 mL). The organic phases were combined, washed twice with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 5-5.

[0116] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ=8.05 (d, J=9.6 Hz, 2H), 7.03 (d, J=9.2 Hz, 2H), 4.45-4.57 (m, 5H), 3.47 (br, 4H), 2.38-2.22 (br, 4H).

Step 4: Synthesis of Compound 5-6

[0117] Under the protection of nitrogen, palladium carbon (0.5 g, 2.25 mmol) was added to a solution of compound 5-5 (4.2 g, 15.95 mmol) in methanol (150 mL) and the nitrogen was replaced with hydrogen by a hydrogen balloon three times. The reaction was stirred at room temperature for 16 hours under hydrogen. The reaction solution was filtered, and the filtrate was concentrated and dried to obtain compound 5-6.

[0118] MS (ESI) m/z:234 [M+H].sup.+.

Step 5: Synthesis of Compound 5-7

[0119] Compound 5-6 (991 mg, 4.25 mmol) and DIEA (686 mg, 5.31 mmol) were added to a solution of compound 1-5 (1.0 g, 3.54 mmol) in isopropanol (10 mL). The reaction solution was stirred at 100° C. for 16 hours. The reaction solution was cooled to room temperature, filtered, and the filter cake was dried to obtain compound 5-7.

Step 6: Synthesis of Compound 5-9

[0120] Compound 5-7 (200 mg, 417 .Math.mol), compound 5-8 (135.3 mg, 417 .Math.mol), potassium carbonate (173 mg, 1.25 mmol) and Pd(dppf)Cl.sub.2 (30.53 mg, 41.72 .Math.mol) were added to 1,4-dioxane (4 mL) and water (5 mL). Under the protection of nitrogen, the reaction solution was stirred at 100° C. for 16 hours. The reaction solution was added with water (30 mL), and extracted three times with ethyl acetate (15 mL). The organic phases were combined, washed twice with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 5-9.

[0121] MS (ESI) m/z:597 [M+H].sup.+.

Step 7: Synthesis of Compound 5

[0122] Tetrabutylammonium fluoride (153 mg, 586.5 .Math.mol) and ethylenediamine (70.5 mg, 1.17 mmol) were added to a solution of compound 5-9 (350 mg, 586.5 .Math.mol) in tetrahydrofuran (10 mL). The reaction solution was stirred at 75° C. for 16 hours. The pH of the reaction solution was adjusted to 8 with sodium hydroxide solution, water (10 mL) was added, and the solution was extracted twice with dichloromethane (10 mL). The organic phases were combined and washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate=10%~100%) to obtain compound 5.

[0123] .sup.1H NMR (400 MHz, DMSO- d.sub.6) δ = 9.44 (s, 1H), 8.74 (d, J=1.0 Hz, 1H), 8.39 (d, J=1.6 Hz, 1H), 8.22 (s, 2H), 8.00 (d, J=9.0 Hz, 3H), 7.75 - 7.68 (m, 2H), 7.63 (d, J=8.5 Hz, 1H), 6.96 (d, J=9.0 Hz, 2H), 4.61 - 4.52 (m, 2H), 4.48 (t, J=6.1 Hz, 3H), 3.20 - 3.03 (m, 4H), 2.42 (br t, J=4.7 Hz, 4H); MS (ESI) m/z: 467 [M+H].sup.+.

Example 6: Preparation of Compound 6

[0124] ##STR00061##

Step 1: Synthesis of Compound 6-2

[0125] Compound 6-1 (338.55 mg, 1.77 mmol) and DIEA (1.14 g, 8.85 mmol) were added to a solution of compound 1-5 (0.5 g, 1.77 mmol) in dimethyl sulfoxide (7 mL). The reaction solution was stirred at 120° C. for 16 hours. After the reaction solution was cooled to room temperature, 50 mL of water was added, a solid was precipitated, filtered, the filter cake was slurried with 10 mL of isopropanol, filtered and dried to obtain compound 6-2.

Step 2: Synthesis of Compound 6-3

[0126] Compound 6-2 (400 mg, 914.63 mmol), compound 5-8 (1.19 g, 3.66 mmol), potassium carbonate (379.23 mg, 2.74 mmol) and Pd(dppf)Cl.sub.2 (66.92 mg, 91.46 .Math.mol) were added to 1,4-dioxane (12 mL) and water (3 mL). Under the protection of nitrogen, the reaction solution was stirred at 100° C. for 16 hours. The reaction solution was added with water (100 mL), and extracted three times with ethyl acetate (150 mL). The organic phases were combined and washed twice with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated by silica gel column chromatography (dichloromethane/methanol, methanol: 0%~10%) to obtain compound 6-3.

[0127] MS (ESI) m/z:555 [M+H].sup.+.

Step 3: Synthesis of Compound 6

[0128] Compound 6-3 (280 mg, 504.72 .Math.mol) was added to dioxane hydrochloride (4 M, 5 mL). The reaction solution was stirred at 25° C. for 16 hours. The pH of the reaction solution was adjusted to 8 with 2 M aqueous sodium hydroxide solution, water (10 mL) was added, and the solution was extracted twice with dichloromethane (10 mL). The organic phases were combined and washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was added with 3 mL of acetone, and the mixture was stirred for 0.5 hours, filtered, and the filter cake was dried to obtain compound 6.

[0129] .sup.1H NMR (400 MHz, DMSO- d.sub.6) δ = 10.80 (br s, 1H), 8.99 (s, 1H), 8.56 (s, 1H), 8.29 (s, 2H), 8.03 (s, 1H), 7.94 (br s, 2H), 7.87 - 7.81 (m, 1H), 7.79 - 7.68 (m, 1H), 7.11 (br d, J=8.9 Hz, 2H), 5.52 (s, 1H), 3.84 (br d, J=11.3 Hz, 3H), 3.51 (br d, J=11.0 Hz, 4H), 2.83 (br d, J=4.4 Hz, 4H); MS (ESI) m/z: 425 [M+H].sup.+.

Example 7: Preparation of Compound 7

[0130] ##STR00062##

Step 1: Synthesis of Compound 7-2

[0131] Compound 7-1 (362.85 mg, 1.77 mmol) and DIEA (1.14 g, 8.85 mmol) were added to a solution of compound 1-5 (0.5 g, 1.77 mmol) in dimethyl sulfoxide (7 mL). The reaction solution was stirred at 120° C. for 16 hours. After the reaction solution was cooled to room temperature, 50 mL of water was added, a solid was precipitated, filtered, the filter cake was slurried with 10 mL of isopropanol, filtered and dried to obtain compound 7-2.

Step 2: Synthesis of Compound 7-3

[0132] Compound 7-2 (123 mg, 272.51 .Math.mol), compound 5-8 (441.87 mg, 1.36 mmol), potassium carbonate (112.99 mg, 817.53 .Math.mol) and Pd(dppf)Cl.sub.2 (19.94 mg, 27.25 .Math.mol) were added to 1 ,4-dioxane (4 mL) and water (1 mL). Under the protection of nitrogen, the reaction solution was stirred at 100° C. for 16 hours. The reaction solution was added with water (50 mL), and extracted three times with ethyl acetate (100 mL). The organic phases were combined and washed twice with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated by silica gel column chromatography (dichloromethane: methanol: 0%~10%) to obtain compound 7-3. MS (ESI) m/z:569 [M+H].sup.+.

Step 3: Synthesis of Compound 7

[0133] Compound 7-3 (127 mg, 223.28 .Math.mol) was added to dioxane hydrochloride (4 M, 5 mL). The reaction solution was stirred at 25° C. for 16 hours. The pH of the reaction solution was adjusted to 8 with 2 M aqueous sodium hydroxide solution, water (10 mL) was added, and the solution was extracted twice with dichloromethane (10 mL). The organic phases were combined and washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated by silica gel column chromatography (dichloromethane/methanol, methanol: 0%~20%) to obtain compound 7.

[0134] .sup.1H NMR (400 MHz, DMSO- d.sub.6) δ = 11.20 (brs, 1H), 9.17 (d, J=1.1 Hz, 1H), 8.68 (s, 1H), 8.37 (s, 2H), 8.19 (s, 1H), 7.96 - 7.68 (m, 2H), 7.46 (s, 1H), 7.33 (s, 1H), 7.20 (s, 1H), 6.96 (br d, J=8.8 Hz, 2H), 3.96 - 3.74 (m, 2H), 3.74 - 3.60 (m, 1H), 3.56 - 3.34 (m, 5H), 3.31 - 3.03 (m, 2H), 2.81 (d, J=4.8 Hz, 3H); MS (ESI) m/z: 439 [M+H].sup.+.

Example 8: Preparation of Compound 8

[0135] ##STR00063##

Step 1: Synthesis of Compound 8-2

[0136] Potassium carbonate (2.94 g, 21.26 mmol), compound 8-1 (1.06 g, 9.21 mmol) were added to a solution of compound 5-1 (1 g, 7.09 mmol) in DMSO (20 mL). The reaction solution was stirred at 100° C. for 16 hours. The reaction solution was concentrated, water (50 mL) was added, and the mixture was extracted three times with ethyl acetate (50 mL). The organic phases were combined, dried, filtered and concentrated to obtain compound 8-2. MS (ESI) m/z:237 [M+H].sup.+.

Step 2: Synthesis of Compound 8-3

[0137] Under the protection of nitrogen, ammonium chloride (1.19 g, 22.22 mmol) and iron powder (2.48 g, 44.44 mmol) were added to a solution of compound 8-2 (1.5 g, 15.95 mmol) in ethanol (8 mL) and water (2 mL). The reaction solution was stirred at 100° C. for 16 hours. The reaction solution filtered and the filtrate was added with water (50 mL), and extracted three times with ethyl acetate (50 mL). The organic phases were combined, dried, filtered, concentrated and dried to obtain compound 8-3. MS (ESI) m/z:207 [M+H].sup.+.

Step 3: Synthesis of Compound 8-4

[0138] Compound 8-3 (401.63 mg, 1.95 mmol) and DIEA (1.14 g, 8.85 mmol) were added to a solution of compound 1-5 (0.5 g, 1.77 mmol) in dimethyl sulfoxide (7 mL). The reaction solution was stirred at 120° C. for 16 hours. After the reaction solution was cooled to room temperature, 50 mL of water was added, a solid was precipitated, filtered, the filter cake was slurried with 10 mL of isopropanol, filtered and dried to obtain compound 8-4.

Step 4: Synthesis of Compound 8-5

[0139] Compound 8-4 (340 mg, 751.64 .Math.mol), compound 5-8 (731.26 mg, 2.25 mmol), potassium carbonate (311.65 mg, 2.25 mmol) and Pd(dppf)Cl.sub.2 (55 mg, 75.16 .Math.mol) were added to 1,4-dioxane (4 mL) and water (1 mL). Under the protection of nitrogen, the reaction solution was stirred at 100° C. for 16 hours. The reaction solution was added with water (50 mL), and extracted three times with dichloromethane (100 mL). The organic phases were combined and washed twice with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated by silica gel column chromatography (dichloromethane/tetrahydrofuran, tetrahydrofuran: 0%~100%) to obtain compound 8-5. MS (ESI) m/z:570 [M+H].sup.+.

Step 5: Synthesis of Compound 8

[0140] Compound 8-5 (350 mg, 614.28 .Math.mol) was added to dioxane hydrochloride (4 M, 5 mL). The reaction solution was stirred at 25° C. for 16 hours. The pH of the reaction solution was adjusted to 8 with 2 M aqueous sodium hydroxide solution, water (10 mL) was added, and the solution was extracted twice with dichloromethane (10 mL). The organic phases were combined and washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated by silica gel column chromatography (dichloromethane/methanol, methanol: 0%~20%) to obtain compound 8.

[0141] .sup.1H NMR (400 MHz, DMSO- d.sub.6) δ = 12.71 - 12.02 (m, 1H), 10.29 (br s, 1H), 8.91 (s, 1H), 8.53 (s, 1H), 8.37 (br d, J=7.5 Hz, 2H), 8.29 (s, 2H), 7.93 (s, 1H), 7.84 (br d, J=8.4 Hz, 3H), 7.80 (s, 1H), 7.77 (s, 1H), 3.24 - 2.97 (m, 1 H), 2.67 (s, 1 H), 1.80 (br s, 3H), 1.49 - 0.33 (m, 6H); MS (ESI) m/z: 440 [M+H].sup.+.

Example 9: Preparation of Compound 9

[0142] ##STR00064##

Step 1: Synthesis of Compound 9-2

[0143] A solution of compound 9-1 (45 g, 189.62 mmol) in methanol (500 mL) and water (200 mL) was cooled to 0° C., and added with sodium periodate (44.61 g, 208.58 mmol). The reaction solution was stirred at 20° C. for 16 hours. The reaction solution was filtered and the filter cake was washed three times with ethyl acetate (50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 9-2.

Step 2: Synthesis of Compound 9-3

[0144] Under the protection of nitrogen, trifluoroacetamide (17.85 g), rhodium acetate (523.43 mg), magnesium oxide (12.73 g), diacetoxyiodobenzene (38.15 g) and potassium carbonate (54.56 g) were added to a solution of compound 9-2 (20 g, 78.95 mmol) in dichloromethane (300 mL). The reaction solution was stirred at 20° C. for 16 hours. The reaction solution was filtered and the filter cake was washed three times with dichloromethane (100 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product which was separated by silica gel column chromatography (petroleum ether/ethyl acetate: ethyl acetate=0%~50%) to obtain compound 9-3.

Step 3: Synthesis of Compound 9-4

[0145] Compound 9-3 (16.0 g) was added to a solution of hydrobromic acid (45%, w/v) in acetic acid (60 mL). The reaction solution was stirred at 20° C. for 10 hours. The pH of the reaction solution was adjusted to 8 with 2 M aqueous sodium hydroxide solution, water (100 mL) was added, and the solution was extracted three times with dichloromethane (150 mL). The organic phases were combined, washed twice with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 9-4.

Step 4: Synthesis of Compound 9-5

[0146] Potassium carbonate (2.94 g, 21.26 mmol), compound 9-4 (1.14 g, 8.50 mmol) were added to a solution of compound 5-1 (1 g, 7.09 mmol) in DMSO (10 mL). The reaction solution was stirred at 100° C. for 16 hours. The reaction solution was concentrated, water (50 mL) was added, and the mixture was extracted three times with ethyl acetate (50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 9-5. MS (ESI) m/z:256 [M+H].sup.+.

Step 5: Synthesis of Compound 9-6

[0147] Under the protection of nitrogen, ammonium chloride (1.05 g, 19.59 mmol) and iron powder (1.09 g, 19.59 mmol) were added to a solution of compound 9-5 (1.0 g, 3.92 mmol) in ethanol (20 mL) and water (5 mL). The reaction solution was stirred at 90° C. for 1 hour. The reaction solution was filtered and the filtrate was added with water (50 mL), and extracted three times with ethyl acetate (50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 9-6. MS (ESI) m/z:226 [M+H].sup.+.

Step 6: Synthesis of Compound 9-7

[0148] Compound 9-6 (0.3 g, 1.33 mmol) and DIEA(516.25 mg, 3.99 mmol) were added to a solution of compound 1-5 (376.18 mg, 1.33 mmol) in dimethyl sulfoxide (10 mL). The reaction solution was stirred at 120° C. for 16 hours. After the reaction solution was cooled to room temperature, 50 mL of water was added, a solid was precipitated, filtered, the filter cake was slurried with 10 mL of isopropanol, filtered and dried to obtain compound 9-7.

Step 7: Synthesis of Compound 9

[0149] Compound 9-7 (200 mg, 424.29 .Math.mol), compound 9-8 (164.66 mg, 848.58 .Math.mol), potassium carbonate (175.92 mg, 1.27 mmol) and Pd(dppf)Cl.sub.2 (31.05 mg, 42.43 .Math.mol) were added to 1,4-dioxane (8 mL) and water (2 mL). Under the protection of nitrogen, the reaction solution was stirred at 100° C. for 16 hours. The reaction solution was added with water (50 mL), and extracted three times with dichloromethane (100 mL). The organic phases were combined and washed twice with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated and purified by high performance liquid chromatography (column: 3-Phenomenex Luna C18 75x30 mm*3 .Math.m; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; B (acetonitrile)%: 45%- 75%, 7 min) to obtain compound 9 (trifluoroacetate).

[0150] .sup.1H NMR (400 MHz, DMSO- d.sub.6): δ = 10.75 (br s, 1H), 9.08 (s, 1H), 8.62 (s, 1H), 8.32 (s, 2H), 8.14 (s, 1H), 7.97 (br d, J=7.5 Hz, 2H), 7.83-7.89 (m, 1H), 7.76-7.81 (m, 1H), 7.18 (br d, J=9.0 Hz, 2H), 4.28 (brd, J=14.6 Hz, 4H), 4.08 (brd, J=13.6 Hz, 2H), 3.76 (br t, J=10.8 Hz, 2H), 3.59-3.67 (m, 2H); MS (ESI) m/z: 459 [M+H].sup.+.

Example 10: Preparation of Compound 10

[0151] ##STR00065##

Step 1: Synthesis of Compound 10-1

[0152] Triethylamine (817.81 .Math.l) and acetyl chloride (209.65 .Math.l) were added to a solution of compound 9-5 (0.5 g, 1.96 mmol) in dichloromethane (10 mL). The reaction solution was stirred at 25° C. for 2 hours. The reaction solution was added with water (50 mL), and extracted three times with dichloromethane (100 mL). The organic phases were combined, washed twice with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 10-1.

Step 2: Synthesis of Compound 10-2

[0153] Under the protection of nitrogen, ammonium chloride (449.76 mg) and iron powder (469.55 mg) were added to a solution of compound 10-1 (0.5 g, 1.68 mmol) in ethanol (20 mL) and water (5 mL). The reaction solution was stirred at 90° C. for 1 hour. The reaction solution was filtered and the filtrate was added with water (50 mL), and extracted three times with ethyl acetate (50 mL). The organic phases were combined, dried, filtered, concentrated and dried to obtain compound 10-2. MS (ESI) m/z:268 [M+H].sup.+.

Step 3: Synthesis of Compound 10-3

[0154] Compound 10-2 (0.2 g) and DIEA (390.91 .Math.l) were added to a solution of compound 1-5 (211.35 mg) in dimethyl sulfoxide (10 mL). The reaction solution was stirred at 120° C. for 16 hours. After the reaction solution was cooled to room temperature, 50 mL of water was added, a solid was precipitated, filtered, the filter cake was slurried with 10 mL of isopropanol, filtered and dried to obtain compound 10-3.

Step 4: Synthesis of Compound 10

[0155] Compound 10-3 (200 mg), compound 9-8 (151.18 mg), potassium carbonate (161.52 mg) and Pd(dppf)Cl.sub.2 (28.50 mg) were added to 1,4-dioxane (8 mL) and water (2 mL). Under the protection of nitrogen, the reaction solution was stirred at 100° C. for 16 hours. The reaction solution was added with water (50 mL), and extracted three times with dichloromethane (100 mL). The organic phases were combined, washed twice with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product which was separated and purified by silica gel column chromatography (dichloromethane/methanol, methanol: 0%~10%) to obtain compound 10.

[0156] .sup.1H NMR (400 MHz, DMSO- d.sub.6): δ = 13.01 (br s, 1H), 9.55 (s, 1H), 8.76 (s, 1H), 8.42 (s, 1H), 8.33 (br s, 1H), 8.09 (br d, J=9.0 Hz, 2H), 7.72-7.78 (m, 2H), 7.64 (d, J=8.5 Hz, 1H), 7.07 (br d, J=9.0 Hz, 2H), 3.87-3.95 (m, 2H), 3.59-3.68 (m, 3H), 3.41 (br d, J=11.3 Hz, 2H), 3.31 (br s, 2H), 1.99 (s, 3H); MS (ESI) m/z: 501 [M+H].sup.+.

Example 11: Preparation of Compound 11

[0157] ##STR00066##

Step 1: Synthesis of Compound 11-2

[0158] Under the protection of nitrogen, compound 1-5 (300 mg, 1.06 mmol) and compound 11-1 (225.85 mg, 1.27 mmol) were mixed and stirred at 120° C. for 16 hours. After the reaction solution was cooled to room temperature, 10 mL of dichloromethane and 2 mL of methanol were added, the mixture was stirred for 15 minutes, filtered, and the filter cake was collected and dried under vacuum to obtain compound 11-2. MS (ESI) m/z:423, 425[M+H].sup.+.

Step 2: Synthesis of Compound 11

[0159] Compound 11-2 (100 mg), compound 9-8 (236 mg), potassium carbonate (97.75 mg) and Pd(dppf)Cl.sub.2 (17.29 mg) were added to 1,4-dioxane (4 mL) and water (1 mL). Under the protection of nitrogen, the reaction solution was stirred at 100° C. for 16 hours. The reaction solution was added with water (20 mL), and extracted three times with dichloromethane (30 mL). The organic phases were combined and washed twice with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated and purified by silica gel column chromatography (dichloromethane/methanol, methanol: 0%~10%) to obtain compound 11.

[0160] .sup.1H NMR (400 MHz, DMSO- d.sub.6): δ = 10.61 (br s, 1H), 9.05 (s, 1H), 8.60 (s, 1H), 8.32 (s, 2H), 8.12 (s, 1H), 8.00 (br d, J=8.1 Hz, 2H), 7.84-7.91 (m, 1H), 7.76-7.82 (m, 1H), 7.33 (br d, J=8.4 Hz, 2H), 5.75 (s, 1H), 3.96 (br d, J=10.5 Hz, 2H), 3.45 (td, J=10.7, 3.6 Hz, 2H), 2.73-2.84 (m, 1H), 1.64-1.78 (m, 4H); MS (ESI) m/z: 411 [M+H].sup.+.

Example 12: Preparation of Compound 12

[0161] ##STR00067##

Step 1: Synthesis of Compound 12-2

[0162] Potassium carbonate (1.47 g), compound 12-1 (585.89 mg) were added to a solution of compound 5-1 (0.5 g) in DMSO (20 mL). The reaction solution was stirred at 100° C. for 16 hours. The reaction solution was concentrated, water (50 mL) was added, and the mixture was extracted three times with ethyl acetate (50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 12-2. MS (ESI) m/z:249 [M+H].sup.+.

Step 2: Synthesis of Compound 12-3

[0163] Under the protection of nitrogen, ammonium chloride (603.26 mg) and iron powder (1.26 g) were added to a solution of compound 12-2 (0.8 g) in ethanol (8 mL) and water (2 mL). The reaction solution was stirred at 100° C. for 16 hours. The reaction solution was filtered and the filtrate was added with water (50 mL), and extracted three times with ethyl acetate (50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 12-3. MS (ESI) m/z:219 [M+H].sup.+.

Step 3: Synthesis of Compound 12-4

[0164] Compound 12-3 (154.53 mg) and DIEA (123.30 .Math.l) were added to a solution of compound 1-5 (200 mg) in isopropanol (2 mL). The reaction solution was stirred at 120° C. for 16 hours. The reaction solution was cooled to room temperature, added with water (50 mL), and extracted three times with ethyl acetate (50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated and purified by silica gel column chromatography (dichloromethane/tetrahydrofuran, tetrahydrofuran: 0%-20%) to obtain compound 12-4. MS (ESI) m/z:464, 466[M+H].sup.+.

Step 4: Synthesis of Compound 12-5

[0165] Compound 12-4 (126.54 mg), compound 5-8 (441.87 mg), potassium carbonate (112.99 mg) and Pd(dppf)Cl.sub.2 (19.94 mg) were added to 1,4-dioxane (8 mL) and water (2 mL). Under the protection of nitrogen, the reaction solution was stirred at 100° C. for 16 hours. The reaction solution was added with water (50 mL), and extracted three times with dichloromethane (100 mL). The organic phases were combined and washed twice with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was separated by silica gel column chromatography (dichloromethane/tetrahydrofuran, tetrahydrofuran: 0%-100%) to obtain compound 12-5. MS (ESI) m/z:582 [M+H].sup.+.

Step 5: Synthesis of Compound 12

[0166] Compound 12-5 (50 mg) was added to dioxane hydrochloride (4 M, 5 mL). The reaction solution was stirred at 25° C. for 16 hours. The pH of the reaction solution was adjusted to 8 with 2 M aqueous sodium hydroxide solution, water (10 mL) was added, and the solution was extracted twice with dichloromethane (10 mL). The organic phases were combined, washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 12.

[0167] .sup.1H NMR (400 MHz, DMSO- d.sub.6) δ = 10.14 (br, 1H), 8.84 - 8.86 (m, 1H), 8.51 (s, 1H), 8.42 (d, J=8.9 Hz, 2H), 8.27 (s, 1H), 7.87 - 7.62 (m, 3H), 7.33 - 7.10 (m, 2H), 6.67 (s, 1H), 5.75 (s, 1H), 3.98-3.91 (m, 4H), 3.71 (s, 4H), 0.98 - 0.68 (m, 4H); MS (ESI) m/z: 452 [M+H].sup.+.

Biological Test Data

Experimental Example 1: In Vitro Test of the Compound’s Inhibitory Effect on Syk Kinase (Enzymatic Experiment)

Experimental Purpose

[0168] The interaction between the substrate and the enzyme was detected by HTRF, using the IC50 value of the compound as an indicator to evaluate the inhibitory effect of the compound on Syk kinase.

Experimental Materials

[0169] Syk kinase (Invitrogen, PV3857) [0170] DTT (Sigma #43815) [0171] ATP (Sigma#A7699) [0172] MgCl.sub.2 (Sigma#63020) [0173] MnCl.sub.2 (Sigma#M1787) [0174] EDTA (Invitrogen#15575-020) [0175] HEPES Buffer (Invitrogen#15630-080) [0176] HTRF®KinEASE™ TK (Cisbio #62TKOPEC, 20000 tests) [0177] Low volume, 384-well, white polystyrene plate (Greiner#784075) [0178] 384 Well Microplates (Greiner #781946) [0179] Centrifuge (Eppendorf #5810R) [0180] Pipette (Eppendorf) [0181] Pipette (Greiner) [0182] Pipette gun (Eppendorf) [0183] Mutidorp Pipette [0184] POD 810 Plate Assembler Automatic Microplate Pretreatment System [0185] Envision Reader

Experimental Steps and Methods

[0186] a) Compound dilution and plate-making: [0187] 1) The compound powder was weighed, dissolved in a certain amount of DMSO at an initial concentration of 10 mM. [0188] 2) The compound was diluted to a concentration of 0.74 mM, and plated with POD18, 135 nL per well, the starting compound concentration was 10 .Math.M, and there were 11 concentration points, with 3-fold descending serial dilutions. [0189] b) Enzyme-substrate reaction stage: [0190] 1) Preparation and dilution of the experimental buffer: the 5×HTRF Buffer in the kit was diluted to 1×, and specified amounts of DTT and MgCl.sub.2 solutions were added for use. [0191] 2) SYK enzyme reaction solution was prepared with 1×HTRF Buffer so that the final reaction concentration of SYK kinase was 0.0156 ng/.Math.L. [0192] 3) TK-Substrate-biotin/ATP mixture was prepared so that the final substrate concentration was controlled at 0.2 .Math.M. ATP concentration was controlled at 2 .Math.M. [0193] 4) The solution and the mixture were added with a Mutidorp pipette, SYK enzyme solution and TK-Substrate-biotin/ATP mixture were each added 5 .Math.L per well, and incubated with 23 solution for 1 hour. [0194] c) Detection stage: [0195] 1) 13.33 mL of ethylenediaminetetraacetic acid solution was added to the detection buffer of the kit, and specified amounts of Eu-labeled antibody and XL-665 were added to prepare the detection solution. [0196] 2) The solution was added with a Mutidorp pipette, 10 .Math.L of detection solution per well, and incubated with 23 solution for 1 hour. The reaction of the enzyme and substrate mixture was stopped. [0197] 3) After centrifugation, the reading was obtained on Envision. [0198] d) Data analyzation: the data was analyzed using XL-Fit and the IC.sub.50 value of the compound was calculated.

Experimental Results

[0199] The experimental results are shown in Table 1.

Experimental Example 2: In Vitro Test of the Compound’s Inhibitory Effect on VEGFR2 Kinase (Enzymatic Experiment)

Experimental Purpose

[0200] The VEGFR2 kinase assay kit is intended for measuring VEGFR2 kinase activity using Kinase-Glo®MAX as a detection reagent, and compound screening and analysis were conducted using the IC.sub.50 value of the compound as an indicator.

Experimental Methods and Procedures

[0201] The ADP-GLO™ Kinase Assay is an analytical reaction in which a luminescent kinase is used to determine the adenosine diphosphate formed by the kinase; ADP is converted to ATP, i.e., an optical signal is generated by Ultra-Glo™ luciferase. The optical signal is positively correlated with ADP quantity and kinase activity. This assay is ideal for determining the activity of the compound, making it ideal for primary screening and kinase selectivity analysis. The ADP-Glo™ Kinase Assay can be used to monitor the activity of virtually any ADP-producing enzyme such as a kinase or ATPase: [0202] 1) The enzyme, substrate, ATP and inhibitor were diluted in a kinase buffer; [0203] 2) 384-well plate: 1 .Math.L of inhibitor or (5% dimethyl sulfoxide solution), 2 .Math.L of KDR enzyme, 2 .Math.L of matrix/ATP mixture; [0204] 3) the mixture was incubated at room temperature for 60 minutes; [0205] 4) 5 .Math.L of ADP-GLO™ reagent was added; [0206] 5) the mixture was incubated for 40 minutes at room temperature; [0207] 6) 10 .Math.L of kinase detection reagent was added; [0208] 7) the mixture was incubated at room temperature for 30 minutes; [0209] 8) the luminescence was recorded (integration time 0.5-1 sec). The data was shown in relative light units (rlu), which was directly related to the amount of ATP produced. There was correlation between the percent of the conversion from ATP to ADP for each kinase amount and the corresponding signal-to-background ratio. [0210] 9) Data analysis: (a) KDR enzyme was titrated with 50 .Math.M of ATP, and the luminescent signal produced by KDR enzyme was shown; (b) staurosporine dose response was generated with 1.5 ng of KDR to determine the IC.sub.50 value of the inhibitor.

[0211] Experimental results: the results are shown in Table 1:

TABLE-US-00001 Experimental data of the inhibition of Syk enzyme and VEFGR2 enzyme by the invention compounds Compound No. IC.sub.50 (nM) for inhibition of Syk enzyme IC.sub.50 (nM) for inhibition of VEFGR2 enzyme Compound 1 15 23 Compound 2 125 31 Compound 5 17 19 Compound 6 50 44 Compound 7 34 42 Compound 8 30 31 Compound 9 23 22 Compound 10 11 41 Compound 11 34 56 Compound 12 315 88

[0212] The results showed: the compound of the invention has significant inhibitory effect on spleen tyrosine kinase (Syk) and vascular endothelial growth factor 2 (VEGFR2).

Experimental Example 3: Activity Inhibition Test of the Compound on 67 Off-Target Kinases (Enzymatic Experiment)

Experimental Purpose

[0213] In order to investigate the specificity of the inhibition of the target by the compound, in this study, the activity and selectivity of test compound 1 on the off-target kinases in related pathways were tested on the Eurofins kinaseProfilerTM platform, and compound screening and analysis were conducted using IC.sub.50 value as the indicator.

Experimental Methods and Procedures

[0214] Compound 1 was detected for each selected kinase using the standard Eurofins Kinase Profiler™ protocol, which followed the relevant standard operating procedures. The IC.sub.50 values of the inhibitory activity of the test compound 1 on 67 kinases including Abl(h), B-Raf(h), BTK(h) and JAK(h) were analyzed.

[0215] Experimental results: the experimental results are shown in Table 2:

TABLE-US-00002 IC.sub.50 of the inhibition of sixty-seven off-target kinases by compound 1 Kinase Name IC.sub.50 (nM) Kinase Name IC.sub.50 (nM) Compound 1 Compound 1 Abl(h) 152 JNK1 α1 (h) 1172 ACTR2(h) >10,000 JNK2α2(h) >10,000 BRK(h) 775 JNK3(h) 279 B-Raf(h) 91 PDGFRα(h) 2702 CDK2/cyclinA(h) >10,000 MAPK1 (h) >10,000 CDK4/cyclinD3(h) >10,000 MAP4K4(h) 63 CDK6/cyclinD3(h) >10,000 MAPKAP-K2(h) >10,000 CSK(h) >10,000 MEK1(h) >10,000 CHK1(h) >10,000 MEK2(h) >10,000 CK1ε(h) 385 MKK4(m) >10,000 FGFR1 (h) 847 MKK6(h) >10,000 FGFR3(h) >10,000 mTOR(h) >10,000 Flt1 (h) 104 mTOR/FKBP12(h) >10,000 Fyn(h) 373 MSK1(h) >10,000 IKKα(h) 946 PKA(h) >10,000 IKKβ(h) >10,000 PKBα(h) >10,000 Flt3(h) 12 ROCK-II(h) 499 Aurora-B(h) 232 Ret(h) 756 BTK(h) 1039 Ros(h) >10,000 c-kit (h) 2843 Rse(h) >10,000 cSRC(h) 367 SAPK2a(h) >10,000 EGFR(h) >10,000 SAPK2b(h) >10,000 ErbB2(h) >10,000 SAPK3(h) >10,000 FAK(h) 469 SAPK4(h) >10,000 FGFR2(h) >10,000 Src(1-530)(h) 827 Hck(h)activated 418 Src(T341 M)(h) 1006 Itk(h) 1849 TGFBR1 (h) >10,000 JAK1 (h) 663 Tie2 (h) 2059 JAK2(h) 141 TrkA(h) 112 JAK3(h) 550 Txk(h) 227 Lck(h) activated 1311 TYK2(h) 117 Lyn(h) 934 Yes(h) 204 IR(h) >10,000

[0216] The results showed: the compound of the invention exhibited very good target specificity, and had no other off-target selectivity problems.

Experimental Example 4: Evaluation of the Compound’s Inhibition of Cytochrome P450 Enzymes In Vitro

Experimental Purpose

[0217] In order to investigate the activity of the compound on human liver microsomal CYP450 enzyme, Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS) method was used in this study to determine the inhibitory effect of compound 1 on five main subtypes (CYP1A2, 2C9, 2C19, 2D6, 3A4) of human hepatocyte CYP450 enzyme, and compound screening and analysis were conducted using IC.sub.50 value as the indicator.

Experimental Methods and Procedures

[0218] 1) A test compound and a standard inhibitor working solution (100x) were prepared; [0219] 2) microsomes were taken out of a -80° C. freezer, thawed on ice, labeled with the date, and put back in the freezer immediately after use; [0220] 3) 20 microliters of matrix solution was added to the corresponding wells; [0221] 4) 20 microliters of phosphate buffered saline was added to the blank wells; [0222] 5) 2 microliters of the test compound and positive control working solution were added to the corresponding wells; [0223] 6) 2 microliters of solvent was added to each of the inhibitor-free wells and the blank wells; [0224] 7) human liver microsome working solution was prepared; [0225] 8) 158 microliters of HLM working solution was added to all wells of the incubation plate; [0226] 9) the plate was heated in a 37° C. water bath for about 10 minutes; [0227] 10) NADPH cofactor solution was prepared; [0228] 11) 20 microliters of NADPH cofactor was added to all of the incubation wells; [0229] 12) all CYPs were subjected to mixed culture in a 37° C. water bath for 10 minutes; [0230] 13) at a certain point in time, the reaction was stopped by adding 400 microliters of cold solution (200 ng/mL of tolbutamide in acetonitrile and 200 ng/mL of Labetalol in acetonitrile); [0231] 14) the samples were centrifuged at 4000 rpm for 20 minutes to precipitate proteins; [0232] 15) 200 microliters of supernatant was transferred into 100 microliters of high performance liquid chromatography water, and shaked for 10 minutes; [0233] 16) LC/MS/MS analysis was started.

Experimental Results

[0234] The experimental results are shown in Table 3:

TABLE-US-00003 CYPs IC.sub.50 (.Math.M) Compound 1 CYP1A2 >50 CYP2C9 >50 CYP2C19 >50 CYP2D6 >50 CYP3A4-M >50

[0235] The results showed: the compound of the invention had no significant inhibitory effect on the five main subtypes (CYP1A2, 2C9, 2C19, 2D6, and 3A4) of CYP450 enzyme.

Experimental Example 5: Pharmacokinetic Evaluation of Ocular Administration to Mouse

Experimental Purpose

[0236] In order to investigate the eye-to-blood ratio of the in vivo exposure amounts of the compound after ocular administration thereof, LC-MS/MS method was used in this study to determine the exposure amounts of compound 1 in cornea and plasma after the ocular administration of the compound 1 to a mouse, where compound screening and analysis were conducted by using the drug concentration at the same time point as an indicator.

Experimental Methods and Procedures

[0237] C57BL/6 mice, weighing 18-20 g, were randomly divided into 4 groups after 3-5 days of acclimatization, with each group including three mice, and the mice were administered by eye drops at 5 .Math.L for each eye.

[0238] The test animals (C57BL/6 mice) were not fasted before administration, but drank water freely before and during the experiment.

[0239] About 0.05 mL of blood was collected from saphenous vein at 1 hour and 3 hours following the eye drop administration, anticoagulated with commercial EDTA-K2, and transferred to a centrifuge for centrifugation. After blood sampling from saphenous vein, the corneas were collected under isoflurane anesthesia. At each of the time points, the plasma and corneas of the mice in the same group were pooled for detection.

[0240] Treatment of plasma and cornea:

[0241] Plasma: within 30 minutes the plasma was transferred to be subject to centrifugation at 4° C., 3200 g for 10 minutes, and 20 .Math.L of plasma was taken from each mouse. At each time point, the plasma of the mice in the same group was pooled for detection.

[0242] Cornea: the cornea was removed, rinsed twice with pre-cooled normal saline, dried with a paper towel, and put into a 1.5 mL centrifuge tube, weighed with an analytical balance by a peeling method, and finally the body weight was weighed. At each time point, the corneas of the mice in the same group were pooled for detection.

[0243] Corneal homogenization method: 1 g of cornea was added to 9 mL of pre-cooled methanol: 15 mM phosphate buffer solution = 1:2, an ultrasonic cell disruptor (No. 2 amplitude rod, amplitude 50%, working for 5 s, stopping for 5 s, and ultrasonic treatment for 3 min) was used.

Experimental Results

[0244] The experimental results are shown in Table 4:

TABLE-US-00004 Drug concentrations in plasma and corneal and eye-to-blood ratio of compound 1 administered to mouse by eye drop Compound 1 Administration dosage 0.1 mg/animal Time point (hour) 1 hour 3 hours Drug concentration in corneal (nmol/kg) 6670 2910 Drug concentration in plasma (nM) 10.4 6.8 eye-to-blood ratio 641 427

[0245] The results showed: the compound of the invention had a high eye-to-blood ratio and was suitable for ocular administration.

Example 6: Pharmacodynamic Test of Mouse Dry Eye Model Induced by Subcutaneous Injection of Scopolamine

Experimental Purpose

[0246] Dry eye syndrome, also known as keratoconjunctivitis sicca, is a common ophthalmic disease, which is mostly caused by ocular surface damage or tear film instability due to abnormal tear quality or quantity or dynamics. Dry eye syndrome is often accompanied by symptoms such as itching, dryness and foreign body sensation in the eyes, which have a serious impact on the vision and quality of life of the patients. The purpose of this experiment is to use mouse dry eye model induced by subcutaneous injection of scopolamine, and administer vehicle and compound 1 to the mice through eye drops, to evaluate the improvement of compound 1 on the dry eye model, and to explore the effective dose of the sample for test.

Experimental Methods and Procedures

[0247] 12 animals were selected from 20 female C57BL/6J mice and divided into two groups according to the tear test results of each animal before the experiment. There were 6 animals per group.

[0248] From the day 1 to day 12 of the experiment, dry eye was induced in mice by subcutaneous injection of scopolamine hydrobromide solution, 4 times/day, 0.1 mL/mouse/time, and the interval between two injections was 3±0.5 h.

[0249] From the 1st to the 12th days of the experiment, the animals were given vehicle and 0.5% compound 1 (i.e., 5 mg/mL) by eye drops, 4 times/day, 3 .Math.L/eye/time, and the interval between two administrations was 3±0.5 h. Please be noted that the modeling agent must be administered before eye drop administration (except for inspections on day 7 and day 12).

[0250] Before modeling, on day 7 and day 12, all experimental animals were subjected to tear test and corneal fluorescent staining score. The tear test on day 7 and day 12 was performed 30 minutes after the second administration, and the corneal fluorescent staining score was performed 30 minutes after the third administration.

[0251] On day 13 of the experiment, the blood samples of the first three animals of the second group were collected at 1 hour after the administration, and then the animals were euthanized, and their eyeballs were collected for collecting the corneas; and the blood samples of the last three animals of the second group were collected 3 hours after the administration, and then the animals were euthanized, and their eyeballs were collected for collecting the corneas.

[0252] The day of modeling was designed as the first day of the experiment.

Drug for Delivery

[0253] 1) A desired amount of test sample was weighed in a suitable container. [0254] 2) A desired volume of 25% (w/v) 2-hydroxypropyl-β-cyclodextrin and 0.2% (v/v) castor oil sterile aqueous solution for injection were added to the above container, and the mixture was stirred until it was visually uniform, and the mixture can be heated and/or sonicated in a 37±2° C. water bath if necessary. The pH was measured and recorded, and the pH was adjusted to be between 7.0 and 7.5 with hydrochloric acid or sodium hydroxide at an appropriate concentration. [0255] 3) Under a laminar flow hood, the prepared test sample was filtered into a sterile container with a 0.22 .Math.m GV filter membrane and then packaged for use.

[0256] The formulated drug for delivery was stored in a refrigerator at 2° C. to 8° C., protected from light, and used within 2 days. At least 30 minutes before administration, the formulated drug for delivery was removed from the refrigerator to reach room temperature.

Molding Preparation

[0257] 1) A desired amount of molding agent was weighed in a suitable container. [0258] 2) A desired volume of normal saline was added to the above container and vortexed until the modeling agent was visually dissolved. [0259] 3) Under a laminar flow hood, the preparation was filtered with a 0.22 .Math.m GV filter membrane into a sterile container.

[0260] The formulated molding preparation was stored in a refrigerator at 2° C. to 8° C., protected from light, and used within 7 days. At least 30 minutes before administration, the formulated molding preparation was removed from the refrigerator to reach room temperature.

Experimental Results

[0261] Tear secretion test: [0262] 1) About 1 g of phenol red powder was weighed in a suitable container; [0263] 2) 20-30 mL of normal saline was added and blended continuously until the solution was saturated; [0264] 3) the saturated solution was filtered through a filter paper into another clean container; [0265] 4) a cotton thread was put into the filtered reagent and soaked for 5-10 min; and [0266] 5) the soaked cotton thread was taken out and dried for later use, and it was used up better within five days.

[0267] Before the formal modeling of the experimental animals, there was an adaptive tear test once a day for at least 2 days.

[0268] A tear secretion test was performed on both eyes of all the experimental animals at least 30 minutes after the second administration before the modeling on day 7 and day 12, respectively. The left eye of each animal was tested first, and after the tear secretion test on the left eyes of all the experimental animals was completed, the right eye of each animal was tested.

[0269] During the test, the phenol red cotton thread was inserted into the lower eyelid near the lateral canthus of the eye to be tested of the mouse using tweezers at one time, while timing for 30 seconds, then the cotton thread was taken out, and the length of the tear trace was measured with a vernier caliper. If the cotton thread fell off during the test, the second measurement was not taken until the animal stood still for 10 minutes or more. The experimental results of the tear secretion on day 7 and day 12 after modeling are shown in FIG. 1.

Corneal Fluorescein Staining

[0270] Corneal fluorescent staining was scored for all experimental animals at least 30 minutes after the third administration before modeling on day 7 and day 12, respectively. Each animal was scored with the left eye first and then the right eye.

[0271] First, about 1 .Math.L of 1% sodium fluorescein was instilled on the cornea of the eye to be tested through a pipette, the upper and lower eyelids of the eye to be tested were closed several times, the cornea was rinsed with normal saline several times, and the corneal fluorescence staining was scored for the eye to be tested using a hand-held slit lamp.

[0272] The scoring standard was: the animal cornea was divided into 5 regions, upper, lower, nasal, temporal and central regions, each region was scored from 0-3 points, and the monocular score was the sum of the scores of the five regions. 0 points, no staining; 1 point, light staining, punctate staining in less than 5 areas; 2 points, moderate staining, punctate staining but no plaque staining; 3 points, heavy staining, obvious fluorescent plaques. For comparison between groups, the mean value of the sum of eye scores in each group was used for comparison. The corneal fluorescent staining score results on day 7 and day 12 after modeling are shown in FIG. 2.

[0273] The results showed: the compound of the invention exhibited statistically significant efficacy in the scopolamine-induced mouse dry eye model.