Heteroindole derivative as well as synthesis method and application thereof

Abstract

The invention relates to the field of pharmacy, in particular to a heteroindole derivative and a synthesis method and application thereof. The heteroindole compound derivative has a structure as shown in a general formula (I), and in the general formula (I), R1 is selected from any one of N-Boc-trans-1, 4-cyclohexanediamine, N-Boc-piperazinyl, trans-1, 4-cyclohexanediamine, piperazinyl and N-methyl piperazinyl; r2 is selected from H or halogen atoms; x, Y and Z are selected from CH or N. Tests prove that the heteroindole derivative has good CDK9 inhibitory activity and selectivity, provides a theoretical basis for treatment of diseases related to the activity or expression quantity of cell cycle-dependent kinase CDK9, and has good medicinal potential; # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of pharmacy, and in particular to a heteroindole derivative and a synthesis method and application thereof. Background Art

[0002] The human genome encodes 20 CDKs, most of which have been identified in their evolutionary relationships. CDKs are divided into two subfamilies: cell cycle-related CDKs (CDK1-7 and 14-18) and transcription-related CDKs (CDK7-13, 19, and 20). Different CDKs interact with different cell cycle proteins to regulate cell cycle stages or perform other functions. Among them, CDK9 is closely related to the occurrence and development of a variety of tumors, and intervention in CDK9 can affect the biological functions of tumor cells. Mechanistically, inhibition of CDK9 can block the phosphorylation of RNAPIICTD, induce downregulation of MYC and Mcl-1 protein levels, and promote apoptosis of tumor cells. Currently, due to the toxicity, lack of specificity, or drug resistance of early-developed CDK9 inhibitors, there are no drugs targeting CDK9 on the market. Therefore, the development of new small molecule inhibitors targeting CDK9 can provide candidate compounds for CDK9-related disease treatment drugs. Summary of the invention

[0003] In order to solve the defects in the prior art, the present invention provides a heteroindole derivative and a synthesis method and application thereof.

[0004] To achieve the above objectives, the specific technical solutions of the present invention are as follows.

[0005] The present invention provides a heteroindole derivative, wherein the heteroindole derivative has a structure as shown in the general formula (I):

[0006]

[0007] Wherein, R1 is selected from any one of N-Boc-trans-1,4-cyclohexanediamino, N-Boc-piperazinyl, trans-1,4-cyclohexanediamino, piperazinyl and N-methylpiperazinyl;

[0008] R2 is selected from H or a halogen atom;

[0009] X, Y, and Z are all selected from CH or N.

[0010] In some embodiments of the present invention, the R1 is preferably trans-1,4-cyclohexanediamino.

[0011] In some embodiments of the present invention, the heteroindole derivative is any one of the following compounds:

[0012]

[0013] The present invention also provides a method for synthesizing the heteroindole derivative, comprising the following steps:

[0014] The heteroindole derivative is obtained by Suzuki coupling reaction of an indole or heteroindole substituted with R3 as shown in formula (1) and a boronic acid or boronic ester intermediate as shown in formula (2), wherein R1 of formula (I) is selected from any one of N-Boc-trans-1,4-cyclohexanediamino, N-Boc-piperazinyl, trans-1,4-cyclohexanediamino, piperazinyl and N-methylpiperazinyl;

[0015] or a Suzuki coupling reaction is performed by the indole or heteroindole substituted by R3 as shown in formula (1) and the boronic acid or boric acid ester intermediate as shown in formula (2) to obtain a Suzuki coupling reaction product as shown in formula (3), and the Suzuki coupling reaction product is further subjected to an acid hydrolysis reaction to remove the protecting group Boc to obtain the heteroindole derivative, wherein R1 of formula (I) is selected from trans-1,4-cyclohexanediamino or piperazinyl;

[0016]

[0017] In the above-mentioned steps of synthesizing heteroindole derivatives,

[0018] The molar ratio of the R3-substituted indole or heteroindole to the boronic acid or boronic acid ester intermediate is 1:1 to 1.5;

[0019] R2 is selected from H or a halogen atom;

[0020] R3 is selected from any one of Cl, Br and I;

[0021] R4 is selected from N-Boc-trans-1,4-cyclohexanediamino or N-Boc-piperazinyl;

[0022] In some embodiments of the present invention, the position of R3 substitution in formula (1) is as shown in formula (4) or formula (5):

[0023]

[0024] In some embodiments of the present invention, the R3 substituted indole is selected from any one of 4-azaindole, 5-azaindole, 7-azaindole and 1,3,7-triazaindole.

[0025] In some embodiments of the present invention, the boronic acid or boronic acid ester intermediate is selected from any one of tert-butyl ((1r,4r)-4-((5-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxin-2-yl)pyridin-2-yl)amino)cyclohexyl)carbamate, (4-(4-(tert-butyloxycarbonyl)piperazin-1-yl)phenyl)boric acid and (4-(4-methylpiperazin-1-yl)phenyl)boric acid.

[0026] In some embodiments of the present invention, the Suzuki coupling reaction comprises: weighing R3-substituted indole or heteroindole and boric acid or boric acid ester intermediate as raw materials, weighing 0.01 to 0.02 times equivalent of [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride of the raw materials as a catalyst, using ethylene glycol dimethyl ether solution as a solvent, dissolving the raw materials and the catalyst in the solvent, adjusting the pH to 8 to 9, and heating at 90 to 120° C. for 6 hours, monitoring the reaction and purifying after completion to obtain the heteroindole derivative;

[0027] The acidolysis reaction comprises: removing the protecting group of the Suzuki coupling reaction product in an organic solvent containing a medium-strong acid to obtain the heteroindole derivative;

[0028] The medium-strong acid is selected from trifluoroacetic acid or hydrogen chloride; the organic solvent is any one of methanol, 1,4-dioxane and dichloromethane; the volume ratio of the medium-strong acid to the organic solvent is 1:2.

[0029] The present invention also provides the use of the heteroindole derivatives or structural analogs of the heteroindole derivatives in the preparation of CDK9 kinase inhibitors, wherein the structural analogs of the heteroindole derivatives include one or more of their pharmaceutically acceptable salts, cocrystals, solvates, esters, acids, metabolites, hydrates, prodrugs and intermediates.

[0030] The present invention also provides the use of the heteroindole derivatives or structural analogs of the heteroindole derivatives in the preparation of drugs for diseases related to the activity of cell cycle-dependent kinase CDK9 or the expression level of cell cycle-dependent kinase CDK9. The structural analogs of the heteroindole derivatives include one or more of their pharmaceutically acceptable salts, cocrystals, solvates, esters, acids, metabolites, hydrates, prodrugs and intermediates.

[0031] In some embodiments of the present invention, the diseases associated with the expression of cell cycle-dependent kinase CDK9 include one or more of breast cancer, liver cancer, colorectal cancer, lung cancer, cervical cancer and brain glioma.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] The present invention designs and synthesizes a new class of heteroindole derivatives, which have good CDK9 inhibitory activity and selectivity, provide a theoretical basis for the treatment of diseases related to the activity or expression of cell cycle-dependent kinase CDK9, and have good medicinal potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is a graph showing the results of a study on the inhibition of CDK9 function by the azaindole compound C6 in Experimental Example 3 of the present invention.

[0035] Figure 2 This is a graph showing the inhibitory experimental results of the azaindole compound C6 on the HCT116 cell subcutaneous transplant tumor model in mice in Experimental Example 4 of the present invention; wherein Figure A is a graph showing the tumor results after the end of drug administration, the upper row is the control group, the lower row is the drug administration group, Figure B is a curve showing the changes in the tumor volume of mice during the drug administration period, Figure C is the average weight of the tumors in each group of mice after the end of drug administration, and Figure D is a curve showing the changes in the body weight of mice during the drug administration period; in Figures B to D, Con represents the control group, and C625 mg / kg represents the drug administration group. DETAILED DESCRIPTION

[0036] The present invention is described in detail below in conjunction with specific examples, but it should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0037] Example 1

[0038] Synthesis of tert-butyl (4-((5-chloro-4-(1H-pyrrolo[2,3-b]pyridin-4-yl)pyridin-2-yl)amino)cyclohexyl)carbamate (C2) and (1r,4r)-N1-(5-chloro-4-(1H-pyrrolo[2,3-b]pyridin-4-yl)pyridin-2-yl)cyclohexane-1,4-diamine (C6)

[0039] (1) Synthesis of the intermediate tert-butyl ((1r,4r)-4-((5-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxin-2-yl)pyridin-2-yl)amino)cyclohexyl)carbamate:

[0040]

[0041] 2-Fluoro-4-iodo-5-chloropyridine (5.14 g, 20 mmol) and N-Boc-1,4-cyclohexanediamine (4.28 g, 20 mmol) were dissolved in 60 mL of N,N-dimethylformamide (DMF), and anhydrous potassium carbonate (K2CO3, 8.28 g, 60 mmol) was added. After nitrogen was replaced in the reaction system, the temperature was raised to 120°C and the reaction was carried out for 4 hours. After TLC detected that the raw materials reacted completely, the heating was stopped, and the reaction solution was poured into 200 mL of ice water with stirring. Solids precipitated, and the filter cake was taken and dried to obtain tert-butyl ((1r, 4r)-4-((5-chloro-4-iodopyridin-2-yl)amino)cyclohexyl)carbamate, a white solid, 8.11 g, and a yield of 90%.

[0042] The white solid (3.61 g, 8 mmol) obtained in the previous step, pinacol borate (2.57 g, 10 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (0.1 mmol), and anhydrous potassium acetate (2.35 g, 24 mmol) were added to 25 mL of dry N,N-dimethylformamide, and the nitrogen was replaced. The temperature was raised to 90°C under nitrogen protection for 4 h. After the reaction of tert-butyl ((1r,4r)-4-((5-chloro-4-iodopyridin-2-yl)amino)cyclohexyl)carbamate was completed, the reaction was stirred for 2 h. The mixture was poured into 100 mL of ice water with stirring, extracted with (3×25 mL) ethyl acetate, the organic phases were combined, and after back-extraction with saturated sodium chloride, the organic phase was added with anhydrous sodium sulfate to dry, and the desiccant was filtered out. After the organic phase was concentrated, silica gel was added to mix the sample, and the mixture was separated and purified by column chromatography on silica gel with petroleum ether: ethyl acetate = 4:1 as the developing solvent to obtain 2.53 g of white powder intermediate tert-butyl ((1r,4r)-4-((5-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxin-2-yl)pyridin-2-yl)amino)cyclohexyl)carbamate, with a yield of 70%.

[0043] (2) Synthesis of tert-butyl (4-((5-chloro-4-(1H-pyrrolo[2,3-b]pyridin-4-yl)pyridin-2-yl)amino)cyclohexyl)carbamate (C2) and (1r,4r)-N1-(5-chloro-4-(1H-pyrrolo[2,3-b]pyridin-4-yl)pyridin-2-yl)cyclohexane-1,4-diamine (C6):

[0044]

[0045] Take a thick-walled pressure bottle, weigh the raw material 4-chloro-7-azaindole (153 mg, 1 mmol) and the intermediate tert-butyl ((1r,4r)-4-((5-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxin-2-yl)pyridin-2-yl)amino)cyclohexyl)carbamate (497 mg, 1.1 mmol) and potassium carbonate (414 mg, 3 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium (30 mg, 0.04 mmol) and dissolve them in 5 mL of solvent (ethylene glycol dimethyl ether: water = 4:1), and add nitrogen. The mixture was replaced with gas for 3 times, and the temperature was raised to 120°C for overnight reaction (6h). After the reaction was completed by TLC detection, the reaction solution was cooled to room temperature and filtered, and the filter cake was washed with a small amount of ethylene glycol dimethyl ether and dried to obtain a crude product. The crude product was dissolved in a dichloromethane:methanol system of 10:1, and silica gel was added to mix the sample. The crude product was separated and purified by column chromatography, and the developing solvent was petroleum ether:ethyl acetate = 1:1 to obtain the compound tert-butyl (4-((5-chloro-4-(1H-pyrrolo[2,3-b]pyridin-4-yl)pyridin-2-yl)amino)cyclohexyl)carbamate (C2), white powder, 225 mg, yield 51%, 1 HNMR(600MHz,DMSO-d6)δ11.85(br s,1H),8.29(d,J=4.95Hz,1H),8.12(s,1H),7.50-7.55(m,1H),7.03(d,J=4.95Hz, 1H),6.76(brt,J=8.53Hz,2H),6.55(s,1H),6.25(dd,J=1.83,3.48Hz,1H),3.61(br s,1H),3.18-3.29(m,1H),1.94-2.03(m,2H),1.80(br d,J=11.19Hz,2H),1.38(s,9H),1.25-1.28(m,2H),1.19-1.24(m,2H).

[0046] The obtained C2 was dissolved in a system with a volume ratio of dichloromethane: trifluoroacetic acid = 2:1 and stirred at room temperature for 2 hours. The dichloromethane and trifluoroacetic acid were evaporated under reduced pressure, and the residual solid was dispersed with dichloromethane. The mixture was treated with triethylamine under stirring until it was alkaline (pH = 8). After the organic phase was concentrated, silica gel was added to mix the sample, and the mixture was separated and purified by column chromatography on silica gel. The developing solvent was dichloromethane: methanol: ammonia = 10:1:0 to dichloromethane: methanol: ammonia = 10:1:0.1 to separate and purify, and (1r,4r)-N1-(5-chloro-4-(1H-pyrrolo[2,3-b]pyridin-4-yl)pyridin-2-yl)cyclohexane-1,4-diamine (C6) was obtained as a white solid. 1H NMR (600MHz, DMSO-d6) δ11.88(br s,1H),8.29(d,J=4.77Hz,1H),8.12(s,1H),7.53(d,J=3.30Hz,1H),7.04(d,J=4.77Hz,1H),6.75(br d,J=7.52Hz,1H),6.55(s,1H),6.25(d,J=3.48Hz,1H),3.62(brs,2H),2.62(br s,1H),1.97(br d,J=9.54Hz,2H),1.81(br d,J=8.44Hz,2H),1.22(brs,2H),1.14-1.19(m,2H).

[0047] Example 2

[0048] Synthesis of 6-(4-(4-methylpiperazin-1-yl)phenyl)-3H-imidazo[4,5-b]pyridine(C10)

[0049]

[0050] Take a thick-walled pressure bottle, weigh 6-bromo-3H-imidazo[4,5-b]pyridine (197 mg, 1 mmol), (4-(4-methylpiperazin-1-yl)phenyl)boric acid (242 mg, 1.1 mmol), potassium carbonate (414 mg, 3 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (30 mg, 0.04 mmol) and dissolve them in 5 mL of solvent (ethylene glycol dimethyl ether: water = 4:1), replace with nitrogen three times, and heat to 90 °C for reaction. Overnight (6 h), after TLC detection, the reaction was completed, and the reaction solution was cooled to room temperature and filtered. The filter cake was washed with a small amount of ethylene glycol dimethyl ether and dried to obtain a crude product. The crude product was dissolved in a dichloromethane:methanol system of 10:1, and silica gel was added to mix the sample. Column chromatography was used for separation and purification. The developing solvent was dichloromethane:methanol = 20:1 to obtain compound 6-(4-(4-methylpiperazin-1-yl)phenyl)-3H-imidazo[4,5-b]pyridine (C10), white powder, 140 mg, yield 48%, 1H NMR(600MHz,DMSO-d6)δ12.94-13.37(m,1H),8.59(br s,1H),8.44(br s,1H),8.22(br s,1H),7.61(brd,J=8.44Hz,2H),7.05(br d,J=8.25Hz,2H),3.20(br s,4H),2.48(br d,J=4.03Hz,4H),2.24(s,3H).

[0051] Example 3

[0052] Synthesis of (1r, 4r)-N1-(5-chloro-4-(1H-indol-4-yl)pyridin-2-yl)cyclohexane-1,4-diamine (C12)

[0053]

[0054] Take a thick-walled pressure bottle, weigh 4-bromo-1H-indole (195 mg, 1 mmol), the intermediate tert-butyl ((1r, 4r)-4-((5-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxin-2-yl)pyridin-2-yl)amino)cyclohexyl)carbamate (497 mg, 1.1 mmol), potassium carbonate (414 mg, 3 mmol) and [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium (30 mg, 0.04 mmol) and dissolve in 5 mL of solvent (ethylene glycol dimethyl ether: water = 4:1) The mixture was replaced with nitrogen three times, and the temperature was raised to 90°C for overnight reaction (6h). After the reaction was completed by TLC detection, the reaction solution was cooled to room temperature and filtered, and the filter cake was washed with a small amount of ethylene glycol dimethyl ether and dried to obtain a crude product; the crude product was dissolved in a dichloromethane:methanol system of 10:1, silica gel was added to mix the sample, and the product was separated and purified by column chromatography with a developing solvent of dichloromethane:methanol = 20:1 to obtain 205 mg of the intermediate tert-butyl ((1r, 4r)-4-((5-chloro-4-(1H-indol-4-yl)pyridin-2-yl)amino)cyclohexyl)carbamate as a white solid with a yield of 47%.

[0055] The intermediate tert-butyl ((1r, 4r)-4-((5-chloro-4-(1H-indol-4-yl)pyridin-2-yl)amino)cyclohexyl)carbamate obtained above was dissolved in a system of dichloromethane:trifluoroacetic acid with a volume ratio of 2:1 and stirred at room temperature for 2 hours. The protecting group was removed to obtain the final product (1r, 4r)-N1-(5-chloro-4-(1H-indol-4-yl)pyridin-2-yl)cyclohexane-1,4-diamine (C12). The purification method and post-treatment are shown in Example 1. 1 H NMR (600MHz, DMSO-d6) δ10.99(s,1H),8.30(d,J=1.28Hz,1H),8.08(s,1H),8.03(d,J=1.83Hz,1H),6.69(m,1H),6 .47-6.60(m,2H),3.49-3.56(m,1H),3.18-3.25(m,1H),1.87-1.94(m,2H),1.76-1.81(m,2H),1.16-1.28(m,4H).

[0056] Example 4

[0057] Synthesis of tert-butyl ((1r,4r)-4-((5-chloro-4-(1H-pyrrolo[2,3-b]pyridin-5-yl)pyridin-2-yl)amino)cyclohexyl)carbamate (C1)

[0058] The synthesis of C1 is similar to the synthesis method of C2 in Example 1, the only difference being that 4-chloro-7-azaindole is replaced by 5-chloro-7-azaindole, and the other raw materials, reaction conditions and post-treatment methods are the same.

[0059] Example 5

[0060] Synthesis of tert-butyl 4-(4-(1H-pyrrolo[2,3-b]pyridin-4-yl)phenyl)piperazine-1-carboxylate (C3)

[0061] The synthesis method of C3 is similar to the synthesis method of C2 in Example 1, the only difference being that the raw materials are replaced by 5-chloro-7-azaindole, and tert-butyl ((1r,4r)-4-((5-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxin-2-yl)pyridin-2-yl)amino)cyclohexyl)carbamate is replaced by (4-(4-(tert-butyloxycarbonyl)piperazin-1-yl)phenyl)boric acid, and the other reaction conditions and post-treatment methods are the same.

[0062] Example 6

[0063] Synthesis of tert-butyl 4-(4-(3H-imidazo[4,5-b]pyridin-6-yl)phenyl)piperazine-1-carboxylate (C4)

[0064] The synthesis of C4 is similar to that of C3 in Example 5, except that 4-chloro-7-azaindole is replaced by 6-bromo-3H-imidazo[4,5-b]pyridine, and other raw materials, reaction conditions and post-treatment methods are the same.

[0065] Example 7

[0066] Synthesis of (1r,4r)-N1-(5-chloro-4-(1H-pyrrolo[2,3-b]pyridin-5-yl)pyridin-2-yl)cyclohexane-1,4-diamine (C5)

[0067] The synthesis method of C5 is similar to the synthesis method of C6 in Example 1, the only difference being that C5 uses C1 as the synthesis raw material, while C6 uses C2 as the synthesis raw material.

[0068] Example 8

[0069] Synthesis of 4-(4-(piperazin-1-yl)phenyl)-1H-pyrrolo[2,3-b]pyridine(C7)

[0070] The synthesis conditions of C7 are similar to the synthesis method of C6 in Example 1, the only difference is that C7 uses C3 as the synthesis raw material, while C6 uses C2 as the synthesis raw material.

[0071] Example 9

[0072] Synthesis of 6-(4-(piperazin-1-yl)phenyl)-3H-imidazo[4,5-b]pyridine(C8)

[0073] The synthesis conditions of C8 are similar to the synthesis method of C6 in Example 1, the only difference is that C8 uses C4 as the synthesis raw material, while C6 uses C2 as the synthesis raw material.

[0074] Example 10

[0075] Synthesis of 4-(4-(4-methylpiperazin-1-yl)phenyl)-1H-pyrrolo[2,3-b]pyridine(C9)

[0076] The synthesis of C9 is similar to the synthesis method of C3 in Example 5, the only difference being that (4-(4-(tert-butyloxycarbonyl)piperazine-1-yl)phenyl)boric acid is replaced by (4-(4-methylpiperazine-1-yl)phenyl)boric acid, and the other raw materials, reaction conditions and post-treatment methods are the same.

[0077] Embodiment 11

[0078] Synthesis of (1r,4r)-N1-(5-chloro-4-(1H-indol-5-yl)pyridin-2-yl)cyclohexane-1,4-diamine (C11)

[0079] The synthesis of C11 is similar to the synthesis method of C12 in Example 3, the only difference being that 4-bromo-1H-indole is replaced by 5-bromo-1H-indole, and the other raw materials, reaction conditions and post-treatment methods are the same.

[0080] The mass spectra and NMR data of compounds C1-C12 are shown in Table 1.

[0081] Table 1 Structures of indole or heteroindole compounds of the present invention and their high-resolution mass spectrometry and nuclear magnetic resonance characterization

[0082]

[0083]

[0084]

[0085] Experimental Example 1

[0086] Cytotoxicity test results of indole or heteroindole compounds of the present invention

[0087] The series of compounds of the present invention were subjected to anti-proliferation experiments on six tumor cells, including human liver cancer cell HepG2, human highly metastatic liver cancer cell MHCC-97H, human liver cancer cell SMMC-7221, human colon cancer cell HCT116, human alveolar adenocarcinoma basal epithelial cell A549 and human liver cancer cell BEL7402, at concentrations of 1 μM and 20 μM. The cell mortality results are shown in Table 2.

[0088] Table 2 Cytotoxicity test results of some indole or heteroindole compounds of the present invention

[0089]

[0090] It can be seen from Table 2 that compounds C1 to C12 have certain killing effects on different tumor cells.

[0091] Experimental Example 2

[0092] The results of inhibition of CDK9 kinase activity by some indole or heteroindole compounds of the present invention

[0093] According to the results of the cell experiment, some compounds with the best activity at the cell level (compounds C5 and C6) were selected to perform an inhibition experiment on CDK9 kinase activity at a concentration of 1 μM / 50 nM, and the experimental results are shown in Table 3. As can be seen from Table 3, it can be seen that compounds C5 and C6 both inhibited CDK9 kinase activity, and C6 had a better inhibitory effect.

[0094] Table 3 Inhibition results of CDK9 kinase activity by compounds C5 and C6

[0095] Compound CDK9 kinase activity (%) @ 1 μM C5 4 C6 1

[0096] Experimental Example 3

[0097] Study on the function of azaindole compound C6 inhibiting CDK9

[0098] CDK9 / cyclinT1 and transcription elongation factor P-TEFb complex form, and CDK9 inhibitors can block the phosphorylation of CDK9 / cyclinT1 on serine 2 in the carboxyl-terminal domain of RNA polymerase II. Therefore, this experimental example tested the phosphorylation level of serine 2 in the carboxyl-terminal domain of RNA polymerase II and the level of parp protein in MDA-MB231 cells treated with compound C6 with the best kinase activity. It was found that compound C6 could inhibit the phosphorylation level of serine 2 and induce parp cleavage. The results are shown in Figure 1 .

[0099] Experimental Example 4

[0100] Inhibitory experiment of the azaindole compound C6 of the present invention on the subcutaneous transplanted tumor model of HCT116 colon cancer cells in mice

[0101] Experimental animals: 10 female BALB / c nude mice aged 4-6 weeks, weighing 16-18 g.

[0102] Construction of HCT116 cell mouse tumor model: HCT116 cell suspension (100 μL, about 10 6 A control group (5% glucose aqueous solution) and a treatment group (25 mg / kg) were set up, and the drug was administered for 14 consecutive days. The weight of mice in each group was recorded every day. After the drug administration, the nude mice were killed, the tumors were removed, photographed and weighed.

[0103] After constructing a common HCT116 cell mouse tumor model, when the tumor initially grows to 50 mm 3 After that, the drug was started. The results of the pharmacodynamic experiment were as follows Figure 2 As shown in the figure, it can be seen that after 14 days of intraperitoneal injection of 25 mg / kg, the tumor volume of mice increased slowly compared with the control group, and the final peeled tumor weight also had a statistical difference. At the same time, the weight changes of mice after administration were also within the normal range.

[0104] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0105] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A heteroindole derivative, characterized in that The heteroindole derivative has a structure as shown in the general formula (I): Wherein, R1 is selected from any one of N-Boc-trans-1,4-cyclohexanediamino, N-Boc-piperazinyl, trans-1,4-cyclohexanediamino, piperazinyl and N-methylpiperazinyl; R2 is selected from H or a halogen atom; X, Y, and Z are all selected from CH or N.

2. The heteroindole derivative according to claim 1, characterized in that The heteroindole derivative is any one of the following compounds:

3. A method for synthesizing the heteroindole derivative according to claim 1, characterized in that: The following steps are involved: The heteroindole derivative is obtained by Suzuki coupling reaction of an indole or heteroindole substituted with R3 as shown in formula (1) and a boronic acid or boronic ester intermediate as shown in formula (2), wherein R1 of formula (I) is selected from any one of N-Boc-trans-1,4-cyclohexanediamino, N-Boc-piperazinyl, trans-1,4-cyclohexanediamino, piperazinyl and N-methylpiperazinyl; or a Suzuki coupling reaction is performed by the indole or heteroindole substituted by R3 as shown in formula (1) and the boronic acid or boric acid ester intermediate as shown in formula (2) to obtain a Suzuki coupling reaction product as shown in formula (3), and the Suzuki coupling reaction product is further subjected to an acid hydrolysis reaction to remove the protecting group Boc to obtain the heteroindole derivative, wherein R1 of formula (I) is selected from trans-1,4-cyclohexanediamino or piperazinyl; In the above-mentioned steps of synthesizing heteroindole derivatives, The molar ratio of the R3-substituted indole or heteroindole to the boronic acid or boronic acid ester intermediate is 1:1 to 1.5; R2 is selected from H or a halogen atom; R3 is selected from any one of Cl, Br and I; R4 is selected from N-Boc-trans-1,4-cyclohexanediamino or N-Boc-piperazinyl; X, Y, and Z are all selected from CH or N.

4. The method for synthesizing the heteroindole derivative according to claim 3, characterized in that: In formula (1), the position of R3 substitution is as shown in formula (4) or formula (5):

5. The method for synthesizing the heteroindole derivative according to claim 4, characterized in that: The R3 substituted indole is selected from any one of 4-azaindole, 5-azaindole, 7-azaindole and 1,3,7-triazaindole.

6. The method for synthesizing heteroindole derivatives according to claim 3, characterized in that: The boronic acid or boronic acid ester intermediate is selected from any one of tert-butyl ((1r, 4r)-4-((5-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxin-2-yl)pyridin-2-yl)amino)cyclohexyl)carbamate, (4-(4-(tert-butyloxycarbonyl)piperazin-1-yl)phenyl)boric acid and (4-(4-methylpiperazin-1-yl)phenyl)boric acid.

7. The method for synthesizing heteroindole derivatives according to claim 3, characterized in that: The Suzuki coupling reaction is specifically as follows: Weighing R3-substituted indole or heteroindole and boric acid or boric acid ester intermediate as raw materials, weighing 0.01 to 0.02 times equivalent of [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride as a catalyst, using ethylene glycol dimethyl ether solution as a solvent, dissolving the raw material and the catalyst in the solvent, adjusting the pH to 8 to 9, heating at 90 to 120° C. for reaction for 6 hours, monitoring the reaction and purifying after completion to obtain the heteroindole derivative; The acidolysis reaction is specifically as follows: The Suzuki coupling reaction product is subjected to removal of the protecting group Boc in an organic solvent containing a medium-strong acid to obtain the heteroindole derivative; The medium-strong acid is trifluoroacetic acid or hydrogen chloride; the organic solvent is any one of methanol, 1,4-dioxane and dichloromethane; the volume ratio of the medium-strong acid to the organic solvent is 1:

2.

8. Use of the heteroindole derivative or the structural analogue of the heteroindole derivative according to claim 1 in the preparation of a CDK9 kinase inhibitor, characterized in that: The structural analogs of the heteroindole derivatives include one or more of their pharmaceutically acceptable salts, cocrystals, solvates, esters, acids, metabolites, hydrates, prodrugs and intermediates.

9. Use of the heteroindole derivative or the structural analogue of the heteroindole derivative according to claim 1 in the preparation of a drug for a disease associated with the activity of cell cycle-dependent kinase CDK9 or the expression level of cell cycle-dependent kinase CDK9, characterized in that: The structural analogs of the heteroindole derivatives include one or more of their pharmaceutically acceptable salts, cocrystals, solvates, esters, acids, metabolites, hydrates, prodrugs and intermediates.

10. The use according to claim 9, characterized in that Diseases associated with the expression of cell cycle-dependent kinase CDK9 include one or more of breast cancer, liver cancer, colorectal cancer and lung cancer.

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