1, 3, 5-trisubstituted indole derivative and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and relates to a 1, 3, 5-trisubstituted indole derivative and application thereof as an ATX inhibitor and in treatment of tumor and fibrosis diseases and promotion of hair growth of mammals. The compounds are compounds as shown in a general formula I and enantiomers, pharmaceutically acceptable salts or prodrugs thereof, and in the general formula I, R1, R2, R3, R4, L, M, Q and n are defined in the claims and the specification. The compound provided by the invention can be used as an ATX inhibitor, is used for treating fibrosis, tumors and other related diseases, and has a potential hair growth promoting function. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine and relates to a 1,3,5-trisubstituted indole derivative and its application as an ATX inhibitor in the treatment of tumors and fibrotic diseases and in promoting hair growth in mammals. Background Art

[0002] Autotaxin (ATX) is a secretory glycoprotein that can hydrolyze LPC (Lysophosphatidylcholine) into choline and lipid signaling molecule LPA (Lysophosphatidate). The ATX-LPA functional axis plays an important role in fibrotic diseases and tumor diseases. Selective inhibition of ATX can effectively control the progression of tumors and fibrosis. In recent years, the research on ATX inhibitors has gradually entered a mature stage. The compound Zirixetastat (WO2014202458A1) entered Phase III clinical research in 2019 for the treatment of idiopathic pulmonary fibrosis (IPF). In addition, drugs such as BBT877 (patent number not disclosed), X-165 (WO2015154023) and PAT-409 (WO2016028686A1) are in Phase I or Phase II clinical research.

[0003] Domestic research on ATX inhibitors is mainly based on simple site modification based on atom replacement (Zirixetastat parent core imidazopyridine structure analogues), that is, the development of Me-too drugs, such as five-membered and six-membered fused ring parent core pyrazolopyridine compounds (WO2019158107, CN202210530649.9), imidazopyridazine compounds (CN202010658085.8), five-membered and five-membered fused ring parent core imidazothiazole compounds (CN202111348160.1), but generally speaking, there are few reports on innovative structural exploration. Previously, 5-thiazolylindole compounds (CN202010043108.4) were reported as ATX inhibitors with novel structures (optimal compound IC 50 The value was less than 1 nM), although it showed excellent anti-fibrotic activity, the activity needs to be further improved.

[0004] Hair loss is an important public health issue. Numerous studies and survey results show that the quality of life and psychological state of patients with hair loss are significantly affected. Hair loss has become a problem for a large number of young people today, and improving the problem of hair loss is urgent. The current clinical methods for improving hair loss mainly include medication and surgical treatment, but both have significant limitations and disadvantages. Although oral and topical medications (such as minoxidil, finasteride, etc.) are relatively inexpensive and can temporarily relieve the symptoms of hair loss, they have many problems such as large side effects, slow onset, unstable efficacy, and recurrence of the disease after discontinuation of medication. Surgical treatment is fast in effect, but the cost is high, and because the surgery uses autologous single hair follicles and hair follicle units for transplantation, the number is limited and the survival rate is unstable. Therefore, it is particularly important to explore new, safe and effective ways to treat hair loss. Summary of the invention

[0005] The present invention aims to provide a 1,3,5-trisubstituted indole derivative and its application as an ATX inhibitor in the treatment of tumors and fibrotic diseases and in promoting hair growth in mammals.

[0006] To achieve the above purpose, the present invention adopts the following technical solution:

[0007] A 1,3,5-trisubstituted indole compound, the compound being a compound represented by general formula I and its enantiomer, pharmaceutically acceptable salt or prodrug:

[0008]

[0009] in,

[0010] L is

[0011] M is

[0012] wherein X is O or N, Y is C or N, and Z is CH, O or N, such that X, Y, Z and the carbon and nitrogen atoms to which they are connected together form a five-membered aromatic heterocyclic ring system;

[0013] Q and W are the same or different and are selected from carbonyl, (C1-C6) straight chain or branched alkyl;

[0014] n is 0 or 1;

[0015] R1 and R2 may be the same or different and independently selected from hydrogen, cyano, hydroxyl, (C3-C8) cycloalkyl, (C1-C6) acyl, (C1-C6) alkoxycarbonyl, (C1-C6) alkyl-substituted sulfone, (C1-C6) alkyl-substituted sulfoxide, (C6-C 10)aryl, 4-10 membered heterocyclic group, 5-10 membered heteroaryl, aminoacyl substituted by 1-2 (C1-C6) alkyl groups, or (C1-C6) straight-chain or branched alkyl, (C2-C6) alkenyl, (C2-C6) alkynyl substituted by 1-3 identical or different substituents as follows; the substituents as follows are selected from hydrogen, halogen, hydroxyl, cyano, (C1-C3) alkoxy, carboxyl, and amino;

[0016] Or R1 and R2 together with the N atom to which they are connected form a 4-10 membered heterocyclic ring containing at least one heteroatom or a 5-10 membered aromatic heterocyclic ring containing at least one heteroatom, wherein the heterocyclic ring and the aromatic heterocyclic ring may be substituted by 1-3 identical or different following groups: halogen, cyano, nitro, hydroxyl, mercapto, carboxyl, (carbon)-oxo, (sulfur)-oxo, (sulfur)-dioxo, (C1-C6)alkyl, methoxy, hydroxy-substituted (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)acyl, (C2-C6)acyloxy, (C3-C8)cycloalkyl, (C1-C6)alkoxycarbonyl;

[0017] R3 is unsubstituted or substituted with at least one of the following substituents: (C1-C6) alkyl, (C2-C6) alkenyl, (C2-C6) alkynyl, (C3-C8) cycloalkyl, (C6-C 10 ) aryl, 5- to 10-membered heteroaryl or 4- to 10-membered heterocyclic group, the following substituents are hydrogen, halogen, hydroxyl, cyano, methoxy, carboxyl, amino;

[0018] R4 is C6~C 10 Aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclic group, unsubstituted or substituted by 1-3 substituents which may be the same or different from the following (C1-C6) alkyl, the following substituents are selected from hydrogen, halogen, C1-C3 alkoxy, C1-C3 alkylthio, cyano, hydroxyl; the aryl, heteroaryl or heterocyclic group may also be optionally substituted independently by 1-4 of the following groups which may be the same or different: halogen, cyano, nitro, hydroxyl, thiol, carboxyl, (C1-C6) alkoxy, (C1-C6) acyl, (C2-C6) acyloxy, (C1-C6) alkoxycarbonyl, (C1-C4) alkyl(oxy)sulfone.

[0019] Preferably, the compound is a compound represented by general formula I and its enantiomer, pharmaceutically acceptable salt or prodrug;

[0020] Where:

[0021] L is

[0022] M is

[0023] wherein X is O or N, Y is C or N, and Z is CH, O or N, so that X, Y, Z and the carbon and nitrogen atoms to which they are connected together form a five-membered aromatic heterocyclic ring system;

[0024] Q and W are the same or different and are selected from carbonyl, (C1-C3) straight chain or branched alkyl;

[0025] n is 0 or 1;

[0026] R1 and R2 may be the same or different and independently represent hydrogen, cyano, hydroxyl, (C3-C6) cycloalkyl, (C1-C4) acyl, (C1-C4) alkoxycarbonyl, (C1-C4) alkyl-substituted sulfone, (C6-C 10 ) aryl, aminoacyl substituted by 1 to 2 (C1 to C4) alkyl groups, (C1 to C4) straight or branched alkyl groups which are unsubstituted or substituted by 1 to 3 identical or different substituents as follows, wherein the substituents are independently selected from hydrogen, halogen, hydroxyl, cyano, methoxy, carboxyl, and amino;

[0027] Or R1 and R2 together with the N atom to which they are attached form a 4-10 membered heterocyclic ring containing at least one heteroatom, and the heterocyclic ring may be substituted by 1-2 identical or different following groups: halogen, cyano, hydroxyl, carboxyl, (carbon)-oxo, (sulfur)-dioxo, (C1-C4)alkyl, methoxy or hydroxy-substituted (C1-C4)alkyl, (C1-C4)alkoxy, (C2-C4)acyloxy, (C1-C4)alkoxycarbonyl;

[0028] R3 is unsubstituted or substituted by 1 to 2 groups independently selected from hydrogen, halogen, hydroxyl, cyano, methoxy, amino groups and the following structures: (C1-C4) straight or branched chain alkyl, (C3-C6) cycloalkyl, phenyl, 5-6 membered heteroaryl or 4-7 membered heterocyclic group;

[0029] R4 is phenyl, naphthyl, 5-6 membered heteroaryl or 4-7 membered heterocyclic group, and the phenyl, naphthyl, heteroaryl or heterocyclic group may be optionally substituted independently by 1 to 3 identical or different following groups: halogen, cyano, nitro, hydroxyl, carboxyl, (C1-C4) alkoxy, (C2-C4) acyl, (C2-C4) acyloxy, (C1-C4) alkoxycarbonyl, unsubstituted or substituted by 1 to 3 (C1-C4) alkyl groups independently selected from hydrogen, halogen, methoxy, methylthio, cyano, hydroxyl groups;

[0030] Further preferably, the compound is a compound represented by general formula I and its enantiomer, pharmaceutically acceptable salt or prodrug;

[0031] Where:

[0032] L is

[0033] M is

[0034] Q is (C1-C3) alkyl;

[0035] n is 0 or 1;

[0036] R1 and R2 may be the same or different and may be selected from hydrogen, (C1-C4) straight or branched alkyl, (C3-C6) cycloalkyl, cyano, hydroxy, hydroxyethyl, cyanoethyl, phenyl, methoxy-substituted (C1-C4) alkyl;

[0037] Or R1 and R2 together with the N atom to which they are attached form a 4-7 membered heterocyclic ring containing at least one heteroatom, and the heterocyclic ring may be substituted by 1-2 identical or different following groups: (C1-C4) straight or branched chain alkyl, (carbon)-oxo, (sulfur)-dioxo, hydroxyl, hydroxymethyl, hydroxyethyl, carboxyl, (C3-C6) cycloalkyl;

[0038] R3 is (C1-C4) alkyl or (C3-C6) cycloalkyl;

[0039] R4 is phenyl or pyridyl, and the phenyl or pyridyl may be optionally independently substituted by 1 to 3 identical or different following groups: halogen, cyano, hydroxyl, carboxyl, (C1-C4) alkyl, C1-C3 hydroxyalkyl, (C1-C4) haloalkyl, methoxy;

[0040] Still further preferably, the compound is a compound represented by general formula I and its enantiomer, pharmaceutically acceptable salt or prodrug;

[0041] Where:

[0042] L is

[0043] M is

[0044] Q is CH2 or n = 0 or 1;

[0045] R1 and R2 may be the same or different and may be selected from hydrogen, hydroxyethyl, methyl, ethyl, cyclopropyl, cyclohexyl, and phenyl;

[0046] Or R1 and R2 together with the nitrogen atom to which they are connected form a 4-6 membered heterocyclic ring containing at least one heteroatom; the heterocyclic ring may also be substituted by at least one C1-C3 alkyl, hydroxyl, C1-C3 hydroxyalkyl, carboxyl, (thio)-dioxo, oxo, etc., including

[0047] R3 is methyl or ethyl;

[0048] R4 is phenyl or pyridyl substituted as follows:

[0049] Still more preferably, the compound is a compound represented by general formula I and its enantiomers, pharmaceutically acceptable salts or prodrugs;

[0050] Where:

[0051] L is

[0052] M is

[0053] Q is CH2 or n = 0 or 1;

[0054] R1 and R2 may be the same or different and may be selected from hydrogen, hydroxyethyl, methyl, ethyl, cyclopropyl, cyclohexyl, and phenyl;

[0055] Or R1 and R2 together with the nitrogen atom to which they are attached form

[0056] R3 is methyl or ethyl;

[0057] R4 is

[0058] More preferably, the compound is the following compound and its enantiomer, pharmaceutically acceptable salt or prodrug;

[0059] 3,4,5-Trichlorobenzyl (S)-(1-ethyl-5-(4-((2-(hydroxymethyl)pyrrolidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate;

[0060] 3,4,5-Trichlorobenzyl (R)-(1-ethyl-5-(4-((2-(hydroxymethyl)pyrrolidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate;

[0061] 3,4,5-Trichlorobenzyl(1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate;

[0062] 3,4,5-Trichlorobenzyl(1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate;

[0063] 3,4-Difluorobenzyl(1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate;

[0064] 2,3-Dichlorobenzyl (1-ethyl-5-(4-(anilinomethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate;

[0065] 3,5-Dichlorobenzyl(1-ethyl-5-(4-((4-methylpiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate;

[0066] 1-(1-ethyl-5-(4-((4-methylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)-3-(4-fluorobenzyl)urea;

[0067] 1-(3-chloro-4-methoxybenzyl)-3-(1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)urea;

[0068] 3,5-Dichlorobenzyl(1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate;

[0069] 4-(Trifluoromethyl)benzyl(1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate;

[0070] 1-(4-chlorobenzyl)-3-(1-ethyl-5-(4-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)urea;

[0071] 3,5-Dichlorobenzyl(1-methyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate;

[0072] (R)-1-(4-methylphenyl)ethyl(5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1-methyl-1H-indol-3-yl)carbamate;

[0073] 1-(4-fluorobenzyl)-3-(1-methyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)urea;

[0074] N-(4-(tert-butyl)phenyl)-2-(1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-2-oxoacetamide;

[0075] N-(4-(tert-butyl)phenyl)-2-(1-ethyl-5-(5-((4-hydroxypiperidin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-2-oxoacetamide;

[0076] N-(4-(tert-butyl)phenyl)-2-(5-(5-((cyclohexyl)methyl)amino)methyl)-1,2,4-oxadiazol-3-yl)-1-ethyl-1H-indol-3-yl)-2-oxoacetamide;

[0077] 3,5-Dimethoxybenzyl(1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate;

[0078] 1-(1-ethyl-5-(5-(tetrahydropyrrol-1-ylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-3-(4-fluorobenzyl)urea;

[0079] 3,5-Dichlorobenzyl(1-ethyl-5-(5-((4-hydroxypiperidin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate;

[0080] 3,5-Dichlorobenzyl (S)-(1-ethyl-5-(5-((2-(hydroxymethyl)tetrahydropyrrol-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate;

[0081] 3,5-Dichlorobenzyl (R)-(1-ethyl-5-(5-((2-(hydroxymethyl)tetrahydropyrrol-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate;

[0082] 1-(1-ethyl-5-(5-((4-hydroxypiperidin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-3-(4-fluorobenzyl)urea;

[0083] 1-((2,4-dichlorobenzyl)oxy)-3-(1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)urea;

[0084] N-((1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamoyl)-4-methylbenzenesulfonamide;

[0085] 3,5-Dichlorobenzyl(1-ethyl-5-(4-(2-oxo-2-(piperidin-1-yl)ethyl)piperazin-1-yl)-1H-indol-3-yl)carbamate;

[0086] 2,3-Dichlorobenzyl(1-ethyl-5-(4-(2-(3-hydroxyazetidin-1-yl)-2-oxoethyl)piperazin-1-yl)-1H-indol-3-yl)carbamate;

[0087] 1-(4-chlorobenzyl)-3-(1-ethyl-5-(4-(2-morpholin-2-oxoethyl)piperazin-1-yl)-1H-indol-3-yl)urea.

[0088] The present invention includes processes for preparing compounds of formula I and intermediates thereof.

[0089] Moreover, according to some common methods in the field to which the present invention belongs, the indole derivatives of general formula I in the present invention can form pharmaceutically acceptable salts with acids. Pharmaceutically acceptable addition salts include inorganic acid and organic acid addition salts, and salts formed with the following acids are particularly preferred: hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, etc. Some derivatives can form pharmaceutically acceptable salts with bases, and salts formed with the following bases are preferred: triethylamine, sodium hydroxide, lithium hydroxide, calcium hydroxide and potassium hydroxide. For example, Example 8 (1-(1-ethyl-5-(4-((4-methylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)-3-(4-fluorobenzyl)urea dihydrochloride) is obtained by reacting the piperazine structure in the structure of Compound 8 with hydrochloric acid to form a salt.

[0090] In addition, the present invention also includes prodrugs of the derivatives of the present invention. The prodrugs of the derivatives of the present invention are indole derivatives of general formula I, which may have weak activity or even no activity themselves, but after administration, they are converted into corresponding biologically active forms under physiological conditions (e.g., by metabolism, solvent decomposition or other means). Among them, the acetyl-protected derivatives are the prodrugs of the corresponding hydrolysis products of the indole derivatives (the corresponding products of Examples 1-3, 10-14 and 21-24 are the acetyl-protected prodrugs thereof).

[0091] General synthetic steps

[0092] The examples and preparations provided below further illustrate and illustrate the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparations is not sufficient to limit the scope of the present invention in any way, but only provides methods for practicing the present invention.

[0093] Those skilled in the art will recognize that the chemical reactions described herein can be used to suitably prepare many other compounds of the invention, and that other methods for preparing the compounds of the invention are considered to be within the scope of the invention. For example, the synthesis of the non-exemplified compounds according to the invention can be successfully accomplished by those skilled in the art by modification methods, such as appropriate protection of interfering groups, by utilizing other known reagents in addition to those described herein, or by making some conventional modifications to the reaction conditions. In addition, the reactions disclosed herein or known reaction conditions are also recognized to be applicable to the preparation of other compounds of the invention.

[0094] The H NMR spectra of the compounds were measured using Bruker ARX-400 / 600, and the mass spectra were measured using Agilent 1100 LC / MS; all reagents used were analytically pure or chemically pure.

[0095] The compounds of the present invention can be synthesized according to the following schemes A to G:

[0096] General Method A:

[0097]

[0098] General Method B

[0099]

[0100] General Method C

[0101]

[0102] General Method D

[0103]

[0104] General Method E

[0105]

[0106] General Method F

[0107]

[0108] General MethodsG

[0109]

[0110] A pharmaceutical composition comprising a compound represented by general formula I and its enantiomers, pharmaceutically acceptable salts or prodrugs.

[0111] An application of the 1,3,5-trisubstituted indole compound, and an application of the compound represented by general formula I and its enantiomer, pharmaceutically acceptable salt or prodrug, or the pharmaceutical composition in the preparation of an ATX inhibitor.

[0112] An application of the 1,3,5-trisubstituted indole compound, the compound represented by the general formula I and its enantiomer, pharmaceutically acceptable salt or prodrug or the pharmaceutical composition in the preparation of a drug for treating or preventing tumors, fibrotic diseases and promoting hair growth in mammals.

[0113] The tumors are mainly breast cancer or glioma, and the fibrotic diseases are mainly pulmonary fibrosis, liver fibrosis or myocardial fibrosis.

[0114] The compound of the present invention has been found to have strong in vitro ATX inhibitory activity through in vitro enzyme activity test experiments. Therefore, it can be used to prepare drugs for treating and / or preventing cancer and fibrotic diseases, such as tumors mainly including breast, lung, liver, kidney, colon, rectum, stomach, prostate, bladder, uterus, pancreas, bone marrow, testis, ovary, lymph, soft tissue, head and neck, thyroid cancer and leukemia, neuroblastoma, etc.; fibrotic diseases mainly include pulmonary fibrosis, liver fibrosis, myocardial fibrosis, renal fibrosis, etc.

[0115] Through the test of inhibiting the proliferation of RAW264.7 cells in vitro, it was found that the compound of the present invention has a good proliferation inhibition effect on RAW264.7 cells and has potential anti-tumor application prospects.

[0116] In vivo experiments preliminarily show that the preferred compounds of the present invention have a good prognostic effect on bleomycin-mediated pulmonary fibrosis tissue, indicating that this series of compounds has great potential as a drug for the treatment of pulmonary fibrosis.

[0117] The active compound of the present invention or its pharmaceutically acceptable salt can be used alone as the only anti-tumor or anti-fibrosis drug, or can be used in combination with the currently marketed anti-tumor / anti-fibrosis drugs (such as pirfenidone, nintedanib, paclitaxel, cisplatin, etc.). Combination therapy is achieved by administering the individual therapeutic components simultaneously, sequentially or separately.

[0118] Pharmacological experiments preliminarily show that the preferred compounds in the present invention have a significant promoting effect on hair regeneration in mice, indicating that this series of compounds has great potential as a drug for the treatment of hair loss.

[0119] The present invention designs and synthesizes a series of 1,3,5-trisubstituted indole derivatives, all of which are new structural types with significant structural novelty. Computational studies have shown that the compounds of the present invention adopt an allosteric binding mode. In vivo and in vitro activity screening has shown that the compounds have significant ATX enzyme inhibitory activity and exhibit excellent anti-tumor activity and fibrosis inhibitory activity. The in vivo and in vitro activities are better than the positive control drugs Zirixetastat and PF-8380, and can provide alternative candidate molecules for the treatment of tumors and fibrotic diseases with obvious advantages, with significant creativity and good application prospects. At the same time, it was found during the experiment that the compounds of the present invention have a significant effect on promoting hair growth in mice, and have great prospects for exploration, development and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0120] Figure 1 This is a diagram showing the improvement effect of compound 3 in Example 3 of the present invention on pulmonary fibrosis tissue.

[0121] Figure 2 This is a schematic diagram of the promoting effect of compound 3 in Example 3 of the present invention on hair growth in mice. DETAILED DESCRIPTION

[0122] The compounds, pharmaceutical compositions and applications of the present invention are further described below in conjunction with the examples.

[0123] The substituents of the compounds prepared in Examples 1 to 29 of the present invention are as follows:

[0124]

[0125] In the general compound structure, M is Q is methylene, n=1, and R3 is ethyl. Specific compounds in Examples 1 to 12 are shown in Table 1.

[0126] Table 1 Examples 1 to 12

[0127]

[0128] In the general compound structure, M is n=1, R3 is methyl. The specific compounds in Examples 13 to 15 are shown in Table 2.

[0129] Table 2 Examples 13 to 15

[0130]

[0131] In the general formula, M is R3 is ethyl, n=1, and the compounds in Examples 16 to 26 are shown in Table 3.

[0132] Table 3 Examples 16 to 26

[0133]

[0134] In the general compound structure, M is Q is methylene, n=1, R3 is ethyl, and the compounds in Examples 27 to 29 are shown in Table 4.

[0135] Table 4 Examples 27 to 29

[0136]

[0137] Example 1: 3,4,5-trichlorobenzyl (1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl) carbamate

[0138] Step 1: Synthesis of 1-ethyl-5-nitro-1H-indole

[0139] At room temperature, add 12g (0.3mol) of 60% NaH to dry DMF (100mL), stir, add 1 (20g, 0.124mol) of DMF solution (80mL) dropwise at 0°C, wait for H2 to escape, add iodoethane (15mL, 0.185mol), and react at room temperature for 2h. After the reaction, add 600mL of saturated NH4Cl solution to a 2L beaker, stir, slowly pour the reaction solution into the beaker for quenching, stir for 20min, filter, and dry the filter cake to obtain 22.8g of yellow solid, with a yield of 97.0%.

[0140] Step 2: Synthesis of 1-(1-ethyl-5-nitro-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one

[0141] At room temperature, 1-ethyl-5-nitro-1H-indole (17.1 g, 0.09 mol) was dissolved in 60 mL of DMF, stirred, and (CF3CO)2O (20.79 g, 0.099 mol) was added dropwise at 0°C. After the addition, the mixture was reacted at room temperature for 2.5 h. After the reaction, 300 mL of water was added to the reaction bottle, stirred, filtered, and the filter cake was dried to obtain 22.13 g of the product with a yield of 86.0%.

[0142] Step 3: Synthesis of 1-ethyl 5-nitro-1H-indole-3-carboxylic acid

[0143] At room temperature, 1-(1-ethyl-5-nitro-1H-indol-3-yl)-2,2,2-trifluoroethane-1-one (16.5 g, 0.0577 mol) was dissolved in 5M NaOH and refluxed for 2 h. After the reaction, 120 mL of water was added to the reaction bottle, stirred, and the pH was adjusted to 3 with HCl. The solid was precipitated, filtered, and the filter cake was dried to obtain 11.5 g of the product with a yield of 85.0%.

[0144] Step 4: Synthesis of 3,5-dichlorobenzyl (1-ethyl-5-nitro-1H-indol-3-yl) carbamate

[0145] At room temperature, 1-ethyl 5-nitro-1H-indole-3-carboxylic acid (10.8 g, 0.046 mol) was added to dry toluene, and 6.5 mL TEA and 7.5 mL DPPA were added under stirring. After 30 min, 3,5-dichlorobenzyl alcohol (12.25 g, 0.069 mol) was added, and the reaction was refluxed for 2 h. After the reaction, the solvent was evaporated, water was added, and a solid was precipitated. After monitoring, the filtrate had no ultraviolet absorption and was discarded. The filter cake was transferred into a beaker and DCM was added and stirred. If there was insoluble matter, it was filtered by suction and the filter cake was dried to obtain 9.5 g of a yellow solid with a yield of 51.1%.

[0146] Step 5: Synthesis of 3,5-dichlorobenzyl (5-amino-1-ethyl-1H-indol-3-yl) carbamate

[0147] At room temperature, 3,5-dichlorobenzyl (1-ethyl-5-nitro-1H-indol-3-yl) carbamate (8.0 g, 0.02 mol) was dissolved in 30 mL MeOH, SnCl2·2H2O (5 eq) was added, and 10 mL concentrated hydrochloric acid was added, and the reaction was refluxed for 3 h. After the reaction, 60 mL of water was added to the reaction bottle, stirred, and the pH was adjusted to 8 with saturated Na2CO3 solution, extracted with EA (60 mL × 3), and the organic layer was washed with saturated brine (60 mL × 3) and dried under vacuum to obtain 3 g of solid, with a yield of 40.2%.

[0148] Step 6: Synthesis of 3,5-dichlorobenzyl (5-azido-1-ethyl-1H-indol-3-yl) carbamate

[0149] At 0°C, add 3,5-dichlorobenzyl (5-amino-1-ethyl-1H-indol-3-yl) carbamate (3g, 0.006mol) to 10mL H2O, stir, add 4.5mL concentrated HCl, and dropwise add NaNO2 (1M) aqueous solution. After 0.5h, dropwise add NaN3 aqueous solution (1M), and react at room temperature for 1.5h. After the reaction, add 30mL water, extract with DCM, evaporate the organic layer to dryness, add n-hexane, stir, filter, and dry the filter cake to obtain 1.60g of the product, with a yield of 66.1%.

[0150] Step 7: Synthesis of 3,5-dichlorobenzyl (1-ethyl-5-(4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl) carbamate

[0151] At 0°C, 3,5-dichlorobenzyl (5-azido-1-ethyl-1H-indol-3-yl) carbamate (1.5 g, 0.004 mol) was dissolved in DMF, stirred, and propargyl alcohol (0.25 g, 0.0045 mol), Vc Na (0.15 g, 0.0009 mol), CuI (0.35 g, 0.002 mol), DIPEA (0.75 g, 0.0074 mol) were added in sequence. After 10 minutes, the mixture was reacted at room temperature for 3 hours. After the reaction, 6 mL of ammonia water was added, stirred, ice water was added, and the mixture was filtered. The filter cake was washed with methanol, filtered, and dried to obtain 1.54 g of the product with a yield of 90.1%.

[0152] Step 8: Synthesis of 3,4,5-trichlorobenzyl(5-(4-(chloromethyl)-1H-1,2,3-triazol-1-yl)-1-ethyl-1H-indol-3-yl)carbamate

[0153] At 0°C, 3,5-dichlorobenzyl (1-ethyl-5-(4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl) carbamate (1.5 g, 0.003 mol) was dissolved in DCM, stirred, and SOCl2 (5 eq) was added dropwise. The mixture was reacted at room temperature for 2 h. The solvent was evaporated, water was added, and the mixture was extracted with DCM. The organic layer was evaporated to dryness to obtain 1 g of the product with a yield of 60.1%.

[0154] Step 9: Synthesis of 3,4,5-trichlorobenzyl (S)-(1-ethyl-5-(4-((2-(hydroxymethyl)pyrrolidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate (Example 1)

[0155] At room temperature, 3,4,5-trichlorobenzyl (5-(4-(chloromethyl)-1H-1,2,3-triazol-1-yl)-1-ethyl-1H-indol-3-yl) carbamate (0.1 g) was dissolved in 4 mL MeCN, stirred, L-prolinol (0.04 g) and potassium carbonate (0.8 g, 3 eq) were added, and the mixture was refluxed for 1.5 h. After the reaction, the solvent was evaporated, water was added and stirred, and the mixture was extracted with DCM. The organic phase was separated, washed with saturated brine, and the organic layer was evaporated to dryness to obtain a crude product. The target compound was obtained after purification by column chromatography with a yield of 20%. MS (ESI) m / z: 577.37 [M+H] + ; 1H NMR (400MHz, DMSO-d6) δ9.32(s,1H),8.67(s,1H),7.89(s,1H),7.78(d,J=8.1Hz,1H),7.74(d,J=9.0Hz,1H),7.54(s,2H),7.1 9(s,1H),5.19(s,2H),4.34(q,J=6.9Hz,2H),3.64(s,2H),2.45(s,4H),1.56-1.47(m,4H),1.38(s,2H),1.30(t,J=7.1Hz,3H).

[0156] According to the method of Example 1, intermediate a9 is used as a raw material to carry out N-alkylation reaction with a small molecular amine to obtain the compound of Example 2-4.

[0157] Example 2: 3,4,5-trichlorobenzyl (R)-(1-ethyl-5-(4-((2-(hydroxymethyl)pyrrolidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate. Analytical data: MS (ESI) m / z: 577.37 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ9.31(s,1H),8.63(s,1H),7.89(s,1H),7.78(d,J=8.1Hz,1H),7.74(d,J=9.0Hz,1H),7.53(s,2H),7.1 9(s,1H),5.19(s,2H),4.34(q,J=6.9Hz,2H),3.64(s,2H),2.45(s,4H),1.56-1.47(m,4H),1.38(s,2H),1.30(t,J=6.9Hz,3H).

[0158] Example 3: 3,4,5-trichlorobenzyl (1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate. Analytical data: Mp: 78.1-79.3°C; MS (ESI) m / z: 577.18 [M+H] + ,575.52[MH] - ; 1H NMR(400MHz,DMSO-d6)δ9.31(s,1H),8.70(s,1H),7.89(s,1H),7.77(s,1H),7.74(s,1H),7.60(s,1H),7.54(s,2H),5.19(s,2H),4.64(s,1H),4 .34(d,J=6.9Hz,2H),3.76(s,2H),3.51(s,1H),2.85(s,2H),2.30(s,2H ), 1.76 (d, J = 13.5Hz, 2H), 1.45 (d, J = 9.1Hz, 2H), 1.30 (t, J = 6.8Hz, 3H). 13 C NMR(100MHz,DMSO-d6)δ155.06,141.63,134.56(2C),132.59(2C),127.86,126.62(2C),123.99,123.22 ,122.91,115.57,111.73,111.07(2C),110.05,64.92(2C),52.60,50.72(2C),39.07,34.14(2C),15.40.

[0159] Example 4: 3,4,5-trichlorobenzyl (1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl) carbamate. Analytical data: Mp: 170.6-171.8°C; MS (ESI) m / z: 563.10 [M+H] + .

[0160] Example 5: 3,4-difluorobenzyl (1-ethyl-5- (4- (morpholinylmethyl) -1H-1,2,3-triazol-1-yl) -1H-indol-3-yl) carbamate

[0161] Step 1: Synthesis of 1-(5-amino-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one

[0162] At room temperature, 1-(1-ethyl-5-nitro-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one (1g, 0.0035mol) was dissolved in THF (8mL), stirred, zinc powder (1.36g) was added, saturated ammonium chloride solution (8mL) was added dropwise, and refluxed for 3h. After the reaction, diatomaceous earth was padded for suction filtration, and the filter cake was washed with EA (30mL) until the yellow color of the filter cake disappeared. The filtrate was transferred to a separatory funnel for separation, and the organic layer was washed with saturated brine and evaporated to dryness to obtain 0.75g of yellow solid, with a yield of 84.3%.

[0163] Step 2: Synthesis of 1-(5-azido-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one

[0164] At room temperature, 1-(5-amino-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one (0.72 g, 0.0028 mol) was added to H2O (8 mL), 37% HCl (2.2 mL) was added, 1M NaNO2 aqueous solution was added dropwise, stirred for 0.5 h, 1M NaN3 aqueous solution was added dropwise, and the reaction was carried out at room temperature for 2 h. After the reaction, 15 mL of water was added, stirred for 10 min, filtered, and the filter cake was dried to obtain 0.54 g of the product with a yield of 68.1%.

[0165] Step 3: Synthesis of 1-(1-ethyl-5-(4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)-2,2,2-trifluoroethane-1-one

[0166] At 0°C, 1-(5-azido-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroethane-1-one (0.52 g, 0.02 mol) was dissolved in DMF (5 mL), stirred, and propargyl alcohol (1.2 eq), Vc Na (0.2 eq), CuI (0.5 eq), and DIPEA (2 eq) were added in sequence. After 10 min, the mixture was reacted at room temperature for 3 h. After the reaction, 4 mL of 5% ammonia water was added, stirred, ice water was added, and the mixture was filtered with suction. The filter cake was dried to obtain 0.54 g of a yellow solid with a yield of 87.0%.

[0167] Step 4: Synthesis of 1-(5-(4-(chloromethyl)-1H-1,2,3-triazol-1-yl)-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one

[0168] At 0°C, 1-(1-ethyl-5-(4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one (0.54 g, 0.0016 mol) was dissolved in DCM (6 mL), thionyl chloride was added dropwise, 1 drop of DMF was added as a catalyst, and the reaction was carried out at room temperature for 2 h. After the reaction, the solvent was evaporated, water was added, and the mixture was extracted with DCM. The organic layer was washed with saturated brine and evaporated to dryness to obtain 0.29 g of solid with a yield of 51.1%.

[0169] Step 5: Synthesis of 1-(1-ethyl-5-(4-(morpholinomethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one

[0170] At room temperature, 1-(5-(4-(chloromethyl)-1H-1,2,3-triazol-1-yl)-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one (0.29 g, 0.0008 mol) was dissolved in acetonitrile (4 mL), stirred, potassium carbonate (5 eq) and morpholine (2 eq) were added, and the mixture was reacted at room temperature for 2 h. After the reaction, the solvent was evaporated, water was added, and the mixture was extracted with DCM. The organic layer was washed with saturated brine and evaporated to dryness to obtain 0.26 g of solid with a yield of 80.1%.

[0171] Step 6: Synthesis of 1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indole-3-carboxylic acid

[0172] At room temperature, 1-(1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one (0.2 g) was added to a reaction bottle, and a 6M sodium hydroxide solution was added, and the temperature was raised to reflux for reaction for 2 h. After the reaction, the pH was adjusted to 3, and a black solid was precipitated. The solid was filtered and monitored to be the product. The liquid still had product residues, and DCM was used for extraction. A white solid precipitated in the water layer, which was filtered and monitored to be the product. The filter cakes were combined and dried to obtain 0.1 g of solid, with a yield of 59.1%.

[0173] Step 7: Synthesis of 3,4-difluorobenzyl(1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate (Example 5)

[0174] At room temperature, 1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indole-3-carboxylic acid (0.1 g) was dissolved in dry toluene, stirred, TEA (2 eq) and DPPA (1.5 eq) were added, and 3,4-difluorobenzyl alcohol (1.5 eq) was added after 30 min, and the temperature was raised to reflux for reaction for 2 h. After the reaction, the solvent was evaporated, water and dichloromethane were added for separation, and the organic layer was washed with saturated brine and evaporated to dryness. The pure product was obtained by column chromatography with a yield of 24.0%. Mp: 151.9~152.7℃; MS (ESI) m / z: 497.04[M+H] + ,518.97[M+Na] + ,495.09[MH] - ; 1H NMR(400MHz,DMSO-d6)δ9.75(s,1H),8.52(s,1H),8.29(s,1H),7.65(s,1H),7.63(s,2H),7.58-7.51(m,1H),7.51-7.44(m,1 H),7.33(s,1H),5.17(s,2H),4.24(q,J=6.9Hz,2H),3.66(s,2H),3.59(t,J=4.1Hz,4H),2.47(s,4H),1.35(t,J=7.1Hz,3H).

[0175] According to the method of Example 5, intermediate b6 is used as a raw material to carry out Curtius rearrangement reaction with benzyl alcohol or benzylamine to obtain the compounds of Examples 6-9 and 15.

[0176] Example 6: 2,3-dichlorobenzyl (1-ethyl-5-(4-(anilinomethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl) carbamate. Analytical data: Mp: 134.6-135.2°C; MS (m / z): 535.21 [M+H] + .

[0177] Example 7: 3,5-dichlorobenzyl (1-ethyl-5-(4-((4-methylpiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate. Analytical data: Mp: 138.6-139.8°C; MS (m / z): 541.21 [M+H] + ,539.16[MH] - .

[0178] Example 8: 1-(1-ethyl-5-(4-((4-methylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)-3-(4-fluorobenzyl)urea dihydrochloride. Analytical data: MS (ESI) m / z: 564.04 [M+H] + .

[0179] Example 9: 1-(3-chloro-4-methoxybenzyl)-3-(1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)urea. Analytical data: MS (ESI) m / z: 524.31 [M+H] + ,546.26[M+Na] + ,522.39[MH] - .

[0180] Example 15: 1-(4-fluorobenzyl)-3-(1-methyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)urea. Analytical data: MS (ESI) m / z: 464.30.46 [M+H] + .

[0181] Example 10: 3,5-dichlorobenzyl (1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate:

[0182] Step 1: Synthesis of 1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indole-3-carboxylic acid

[0183] The intermediate c1 of the C series compounds was prepared by a method similar to that of the general method B for 1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indole-3-carboxylic acid (b6).

[0184] Step 2: Synthesis of 5-(4-((4-acetoxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1-ethyl-1H-indole-3-carboxylic acid

[0185] At room temperature, 1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indole-3-carboxylic acid (2g), acetic anhydride (2.2eq), TEA (2.2eq), and DMAP (0.1eq) were added to 15mL DCM in sequence and reacted at room temperature for 2h. After the reaction, water was added to separate the liquid from DCM, and the organic layer was evaporated to dryness to obtain 1.22g of yellow solid, with a yield of 55%.

[0186] Step 3: Synthesis of 1-((1-(3-((((3,5-dichlorobenzyl)oxy)carbonyl)amino)-1-ethyl-1H-indol-5-yl)-1H-1,2,3-triazol-4-yl)methyl)piperidin-4-yl acetate

[0187] At room temperature, 5-(4-((4-acetoxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1-ethyl-1H-indole-3-carboxylic acid (0.2 g) was dissolved in dry toluene, stirred, TEA (2 eq) and DPPA (1.5 eq) were added, and 3,4-difluorobenzyl alcohol (1.5 eq) was added after 30 min, and the temperature was raised to reflux for reaction for 2 h. After the reaction, the solvent was evaporated, water and dichloromethane were added for separation, the organic layer was washed with saturated brine, and evaporated to dryness to obtain 0.15 g of solid, with a yield of 54.0%.

[0188] Step 4: Synthesis of 3,5-dichlorobenzyl(1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate (Example 10)

[0189] At room temperature, 1-((1-(3-((((3,5-dichlorobenzyl)oxy)carbonyl)amino)-1-ethyl-1H-indol-5-yl)-1H-1,2,3-triazol-4-yl)methyl)piperidin-4-yl acetate (0.1 g) was dissolved in 4 mL of dioxane, and 1.5 M NaOH solution was added under stirring and reacted at 40°C. After the reaction, water was added, and the mixture was separated from DCM. The organic layer was washed with saturated brine and evaporated to dryness to obtain a crude product. The pure product was purified by column chromatography with a yield of 23%. Mp: 70.8-71.7°C; MS (m / z): 543.00 [M+H] + ,541.08[MH] - ; 1 H NMR(400MHz,DMSO-d6)δ9.80(s,1H),8.49(s,1H),8.30(s,1H),7.66(s,1 H),7.63(s,2H),7.60(s,1H),7.53(s,2H),5.20(s,2H),4.56(s,1H),4.28 -4.19(m,2H),3.62(s,2H),3.45(s,1H),2.76(d,J=10.9Hz,2H),2.63(s,2 H), 2.12 (t, J = 10.0Hz, 2H), 1.72 (d, J = 10.3Hz, 2H), 1.35 (t, J = 7.0Hz, 3H).

[0190] According to the method of Example 10, the compounds of Examples 11-14 were prepared.

[0191] Example 11: 4-(Trifluoromethyl)benzyl(1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate. Analytical data: MS (ESI) m / z: 543.06 [M+H] + ,541.08[MH] - .

[0192] Example 12: 1-(4-chlorobenzyl)-3-(1-ethyl-5-(4-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)urea. Analytical data: MS (m / z): 537.17 [M+H]+ .

[0193] Example 13: 3,5-dichlorobenzyl (1-methyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate. Analytical data: MS (ESI) m / z: 528.98 [M+H] + .

[0194] Example 14: (R)-1-(4-methylphenyl)ethyl (5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1-methyl-1H-indol-3-yl)carbamate. Analytical data: MS (ESI) m / z: 489.10 [M+H] + , 487.15[MH] - ,511.09[M+Na] + .

[0195] Example 16: N-(4-(tert-butyl)phenyl)-2-(1-ethyl-5-(5-(morpholinomethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-2-oxoacetamide

[0196] Step 1: Synthesis of 2-(5-cyano-1-ethyl-1H-indol-3-yl)-2-oxoacetyl chloride

[0197] Oxalyl chloride (2.4 eq) was dissolved in 40 mL of dry ether at -10°C, 1-ethyl-5-cyanoindole (4 g) was added thereto, and the mixture was stirred at room temperature for 2 h and monitored by TLC. After the reaction, the solvent was evaporated to obtain 6.12 g of a light yellow solid with a yield of 100%.

[0198] Step 2: Synthesis of N-(4-(tert-butyl)phenyl)-2-(5-cyano-1-ethyl-1H-indol-3-yl)-2-oxoacetamide

[0199] 2-(5-cyano-1-ethyl-1H-indol-3-yl)-2-oxoacetyl chloride (1.42 g) was pre-dissolved in 15 mL of dry DCM, DIPEA (2 eq) was added, and finally the mixture was slowly dripped into a solution of 4-(tert-butyl)aniline (1.5 eq) pre-dissolved in dry DCM, and stirred at room temperature for 2 h after the dripping, and monitored by TLC. After the reaction, the mixture was first washed with 0.5 M HCl, the organic phase was separated, and then washed with saturated NaHCO3, and the organic layer was evaporated to obtain 1.83 g of brown solid, with a yield of 89.7%.

[0200] Step 3: Synthesis of N-(4-(tert-butyl)phenyl)-2-(1-ethyl-5-(N'-hydroxycarbamoyl)-1H-indol-3-yl)-2-oxoacetamide

[0201] N-(4-(tert-butyl)phenyl)-2-(5-cyano-1-ethyl-1H-indol-3-yl)-2-oxoacetamide (1.8 g) was dissolved in EtOH, hydroxylamine hydrochloride (1.5 eq) and TEA (2 eq) were added in sequence, the temperature was raised to 70°C, the reaction was continued overnight, and TLC was monitored. After the reaction, part of the solvent was evaporated, and solid was precipitated by cooling in cold hydrazine. 1.02 g of white solid was obtained by suction filtration, and the yield was 52.3%.

[0202] Step 4: Synthesis of N-(4-(tert-butyl)phenyl)-2-(5-(5-(chloromethyl)-1,2,4-oxadiazol-3-yl)-1-ethyl-1H-indol-3-yl)-2-oxoacetamide

[0203] N-(4-(tert-butyl)phenyl)-2-(1-ethyl-5-(N'-hydroxycarbamoyl)-1H-indol-3-yl)-2-oxoacetamide (1 g) was dissolved in 10 mL of DMF, chloroacetyl chloride (1.2 eq) was added dropwise, the temperature was raised to 100°C, the reaction was continued for 4 h, and TLC was monitored. After the reaction was completed, water was added and stirred, and 0.95 g of a brown-yellow solid was obtained by suction filtration, with a yield of 83.0%.

[0204] Step 5: Synthesis of N-(4-(tert-butyl)phenyl)-2-(5-(5-((diethylamino)methyl)-1,2,4-oxadiazol-3-yl)-1-ethyl-1H-indol-3-yl)-2-oxoacetamide (Example 16)

[0205] N-(4-(tert-butyl)phenyl)-2-(5-(5-(chloromethyl)-1,2,4-oxadiazol-3-yl)-1-ethyl-1H-indol-3-yl)-2-oxoacetamide (0.15 g) was dissolved in 4 mL of isopropanol, diethylamine (1.2 eq) was added thereto, and the mixture was refluxed for 12 h and monitored by TLC. After the reaction, water was added and stirred, and DCM was extracted and evaporated to dryness to obtain a crude product, which was purified by column chromatography to obtain a pure product with a yield of 21.2%. Mp: 204.7-206.8 °C; MS (m / z): 515.60 [M+H] + ; 1H NMR (400MHz, DMSO-d6) δ10.64(s,1H),9.04(d,J=1.4Hz,1H),8.95(s,1H),8.01(dd,J=8.6,1.6Hz,1H),7.87(d,J=8.6Hz,1H),7.78(d,J=8.7 Hz,2H),7.40(d,J=8.7Hz,2H),4.44(q,J=7.2Hz,2H),4.02(s,2H),3.65-3.61(m,4H),2.63-2.58(m,4H),1.46(t,J=7.2Hz,3H),1.29(s,9H).

[0206] According to the method of Example 16, the compounds of Examples 17-18 were prepared.

[0207] Example 17: N-(4-(tert-butyl)phenyl)-2-(1-ethyl-5-(5-((4-hydroxypiperidin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-2-oxyacetamide. Analytical data: Mp: 85.7-87.8°C; MS (m / z): 530.28 [M+H] + ,552.41[M+Na] + .

[0208] Example 18: N-(4-(tert-butyl)phenyl)-2-(5-(5-((cyclohexyl)methyl)amino)methyl)-1,2,4-oxadiazol-3-yl)-1-ethyl-1H-indol-3-yl)-2-oxyacetamide. Analytical data: Mp: 161.7-162.8°C; MS (ESI) m / z: 542.27 [M+H] + .

[0209] Example 19: 3,5-dichlorobenzyl (1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate

[0210] Step 1: Synthesis of 1-ethyl-3-(2,2,2-trifluoroacetyl)-1H-indole-5-cyano

[0211] 5-Cyanoindole (5.22 g) and TEA (1.5 eq) were dissolved in 25 mL DMF, and trifluoroacetic anhydride (2 eq) was added dropwise under ice bath conditions. After the addition was complete, the mixture was reacted at room temperature for 3 h. After the reaction, solids were precipitated, which were filtered and dried to obtain 6.12 g of pure white solids, with a yield of 90.3%.

[0212] Step 2: Synthesis of 1-ethyl-N'-hydroxy-3-(2,2,2-trifluoroacetyl)-1H-indole-5-carboximide

[0213] 1-Ethyl-3-(2,2,2-trifluoroacetyl)-1H-indole-5-cyano (3.37 g), hydroxylamine hydrochloride (1.5 eq), TEA (2 eq) were dissolved in ethanol in turn and refluxed for 4 h. After the reaction, 20 mL of water was added and the ethanol was evaporated to remove a large amount of white solids. The solids were filtered and the pH of the filtrate was adjusted to 5.5-6. Solids were precipitated. The filter cakes were combined to obtain 3.2 g of solids with a yield of 85.1%.

[0214] Step 3: Synthesis of 1-ethyl-5-(N'-hydroxycarbamoyl)-1H-indole-3-carboxylic acid

[0215] 1-Ethyl-N'-hydroxy-3-(2,2,2-trifluoroacetyl)-1H-indole-5-carboximide (3 g) was added to 30% sodium hydroxide solution and refluxed for 3 h. After the reaction, the solution was cooled to room temperature and the pH was adjusted to 4.5-5 to precipitate an off-white solid, which was filtered and dried to obtain a filter cake of 2.05 g with a yield of 83.0%.

[0216] Step 4: Synthesis of 5-(5-(chloromethyl)-1,2,4-oxadiazol-3-yl)-1-ethyl-1H-indole-3-carboxylic acid

[0217] 5-(5-(Chloromethyl)-1,2,4-oxadiazol-3-yl)-1-ethyl-1H-indole-3-carboxylic acid (1.5 g) and chloroacetyl chloride (1.2 eq) were dissolved in 15 mL DMF and reacted at 90° C. for 2 h. After the reaction, the solution was cooled to room temperature and water was added to precipitate a light gray solid, which was filtered and dried to obtain 1.38 g of an off-white solid with a yield of 75%.

[0218] Step 5: Synthesis of 1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indole-3-carboxylic acid

[0219] 5-(5-(chloromethyl)-1,2,4-oxadiazol-3-yl)-1-ethyl-1H-indole-3-carboxylic acid (0.5 g), morpholine (1.2 eq) and potassium carbonate (2 eq) were added to acetonitrile and refluxed for 30 min. After the reaction, the solvent was evaporated, water was added, and the pH was adjusted to 4.5-5, and 0.42 g of white solid was precipitated, with a yield of 72.1%.

[0220] Step 6: Synthesis of 3,5-dimethoxybenzyl(1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate (Example 19)

[0221] 1-Ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indole-3-carboxylic acid (0.1 g) was added to a mixture of TEA (2 eq) and DPPA (2 eq) in dry toluene and reacted at room temperature for 30 min. 3,5-dimethoxybenzyl alcohol (1.2 eq) was then added and the temperature was raised to reflux for 1 h. After the reaction was completed, toluene was removed under reduced pressure, water was added, and DCM was used for extraction. The crude product was evaporated to dryness and purified by column chromatography to obtain a pure product with a yield of 18%. MS (ESI) m / z: 522.17 [M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ9.92(s,1H),8.60(s,1H),7.79(dd,J=8.7,1.3Hz,1H),7.63(s,1H),7.59(d,J=8.7Hz,1H),6.62(s,2H),6.4 7(s,1H),5.12(s,2H),4.22(q,J=7.1Hz,2H),3.96(s,2H),3.76(s,6H),3.64-3.59(m,4H),2.59-2.55(m,4H),1.35(t,J=7.2Hz,3H).

[0222] According to the method of Example 19, the compounds of Examples 20-24 were prepared.

[0223] Example 20: 1-(1-ethyl-5-(5-(tetrahydropyrrol-1-ylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-3-(4-fluorobenzyl)urea. Analytical data: MS (ESI) m / z: 463.12 [M+H] + .

[0224] Example 21: 3,5-dichlorobenzyl (1-ethyl-5-(5-((4-hydroxypiperidin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate. Analytical data: Mp: 168.7-169.8°C; MS (m / z): 544.20 [M+H] + .

[0225] Example 22: 3,5-dichlorobenzyl (S)-(1-ethyl-5-(5-((2-(hydroxymethyl)tetrahydropyrrol-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate. Analytical data: MS (ESI) m / z: 565.59 [M+Na] + ,542.04[MH] - .

[0226] Example 23: 3,5-dichlorobenzyl (R)-(1-ethyl-5-(5-((2-(hydroxymethyl)tetrahydropyrrol-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate. Analytical data: MS (m / z): 565.59 [M+Na] + .

[0227] Example 24: 1-(1-ethyl-5-(5-((4-hydroxypiperidin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-3-(4-fluorobenzyl). Analytical data: Mp: 176.8~177.8℃; MS (m / z): 493.01[M+H] + ,515.06[M+Na] + .

[0228] Example 25: 1-((2,4-dichlorobenzyl)oxy)-3-(1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)urea

[0229] 1-Ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indole-3-carboxylic acid (0.1 g) was added to a mixture of TEA (2 eq) and DPPA (2 eq) in dry toluene and reacted at room temperature for 30 min. 2,4-dichlorobenzylhydroxylamine (1.2 eq) was then added and the temperature was raised to reflux for 1 h. After the reaction was completed, toluene was evaporated under reduced pressure, water was added, and DCM was used for extraction. The crude product was purified by column chromatography to obtain a pure product with a yield of 33.2%. Analytical data: MS (ESI) m / z: 567.19 [M+Na] + ,542.87[MH] - .

[0230] Example 26: N-((1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamoyl)-4-methylbenzenesulfonamide

[0231] 1-Ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indole-3-carboxylic acid (0.1 g) was added to a mixture of TEA (2 eq) and DPPA (2 eq) in dry toluene and reacted at room temperature for 30 min. Then 4-methylbenzenesulfonamide (1.2 eq) was added and the temperature was raised to reflux for 1 h. After the reaction was completed, toluene was evaporated under reduced pressure, water was added, and DCM was used for extraction. The crude product was purified by column chromatography to obtain a pure product with a yield of 27.1%. Analytical data: MS (ESI) m / z: 525.27 [M+H] + ,547.16[M+Na]+ ,523.28[MH] - .

[0232] Example 27: 3,5-dichlorobenzyl (1-ethyl-5- (4- (2-oxo-2- (piperidin-1-yl) ethyl) piperazin-1-yl) -1H-indol-3-yl) carbamate

[0233] Step 1: 5-Bromo-1-ethyl-1H-indole (g2)

[0234] At room temperature, add 50.1g (2.5mol) of 60% NaH to 500mL of DMF, stir, slowly heat to 40°C, add 194.97g (1.0mol) of 5-bromoindole, wait for H2 to escape, cool to 25°C, add 234.0g (1.5mol) of iodoethane, and react at room temperature for 2h. After the reaction, add 800mL of water, stir for 30min, and filter to obtain 195.1g of solid, with a yield of 87.5%.

[0235] Step 2: 1-(5-Bromo-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroacetyl-1-one (g3)

[0236] At room temperature, 89.2g (0.4mol) of the intermediate 1-(5-bromo-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroacetyl-1-one (g2) was dissolved in 400mL DMF, stirred, and 83mL (0.6mol) of trifluoroacetic anhydride was added dropwise under ice bath conditions. After the addition, the mixture was reacted at room temperature for 1.5h. After the reaction, the mixture was poured into 2500mL of water, stirred for 30min, and then filtered to obtain 98.5g of solid, with a yield of 78.4%.

[0237] Step 3: 5-Bromo-1-ethyl-1H-indole-3-carboxylic acid (g4)

[0238] At room temperature, 80.1 g (0.25 mol) of the intermediate g3 was added to 4 M NaOH solution (600 mL) and refluxed for 2 h. After the reaction, the mixture was cooled to room temperature, the pH was adjusted to 5 with 6 M HCl, and the mixture was filtered and dried to obtain 59 g of solid with a yield of 79.1%.

[0239] Step 4: 5-(4-(tert-Butyloxycarbonyl)piperazin-1-yl)-1-ethyl-1H-indole-3-carboxylic acid (g5)

[0240] At room temperature, 67.6g (0.25mol) of intermediate g4 was dissolved in 300mL of dry toluene, and then 34.6g of N-Boc piperazine (0.37mol), 48.2g of sodium tert-butoxide (0.5mol), 7.5g of JohnPhos ligand (0.025mol) were added to the solution in sequence under stirring, and finally 10.98g of Pd2(dba)3 (0.012mol) was added. Under N2 protection, the mixture was heated to 115°C for 2h. After the reaction was completed by TLC monitoring, the mixture was filtered with diatomaceous earth pads, and the diatomaceous earth was rinsed with ethyl acetate. After the filtrate was evaporated to dryness, 300mL of n-hexane was added, and after stirring for 30min, the mixture was filtered with suction, and the filter cake was washed with n-hexane (3×60mL) and discarded. The n-hexane layer was evaporated to dryness to obtain 73.3g of solid, with a yield of 85.6%.

[0241] Step 5: tert-Butyl 4-(3-((((3,5-dichlorobenzyl)oxy)carbonyl)amino)-1-ethyl-1H-indol-5-yl)piperazine-1-carboxylate (g6)

[0242] In a 1000mL round-bottom flask, 56g (0.15mol) of intermediate g5 was dissolved in 300mL of dry toluene, and then 45.5g of triethylamine (0.45mol) and 49.5g (0.18mol) of DPPA were added to the solution. After stirring and reacting at 80°C for 30min, 31.9g (0.18mol) of 3,5-dichlorobenzyl alcohol was added, and the mixture was stirred and reacted for 2h under reflux. After the reaction was completed by TLC monitoring, toluene was evaporated, 500ml of water was added, and the mixture was filtered after stirring. The liquid layer was discarded, and the upper solid was added to 300ml of acetonitrile, stirred, and filtered. The filter cake was washed with acetonitrile (3×100mL) and dried to obtain 54.2g of solid, with a yield of 66.2%.

[0243] Step 6: 3,5-Dichlorobenzyl (1-ethyl-5-(piperazin-1-yl)-1H-indol-3-yl) carbamate (g7)

[0244] At room temperature, add 120mL of HCl in dioxane solution (4M) to a 500mL eggplant-shaped bottle, slowly add 10.9g (0.02mol) of intermediate g6 in batches under stirring, and stir for 2h at room temperature. After the reaction is completed by TLC monitoring, dioxane is evaporated under reduced pressure, 500ml of water is slowly added under stirring, and extracted with 200mL of dichloromethane, the organic layer is discarded, the aqueous layer is adjusted to pH=10 with 2N sodium hydroxide solution, extracted with dichloromethane (3×150mL), the organic layers are combined, washed with saturated brine and water in turn, dried over anhydrous sodium sulfate, and evaporated to dry dichloromethane to obtain 8.5g of solid, with a yield of 95.1%.

[0245] Step 7: 3,5-Dichlorobenzyl(1-ethyl-5-(4-(2-oxo-2-(piperidin-1-yl)ethyl)piperazin-1-yl)-1H-indol-3-yl)carbamate (Example 27)

[0246] In a 50mL eggplant-shaped bottle, 0.89g g7 (0.002mol) and 0.85g chloroacetylpiperidine (0.005mol) were dissolved in 8mL acetonitrile, and 0.69g potassium carbonate (0.005mol) was added under stirring, and the mixture was refluxed under stirring for 3h. After the reaction was completed by TLC monitoring, potassium carbonate was removed by suction filtration, acetonitrile was evaporated under reduced pressure, the residue was dissolved in 20mL dichloromethane, extracted with 4M hydrochloric acid (2*15mL), the organic layer was discarded, the aqueous layer was adjusted to pH=10 with 6N sodium hydroxide solution, extracted with dichloromethane (3*10mL), the organic layers were combined, washed with saturated brine and water in turn, dried over anhydrous sodium sulfate, and dichloromethane was evaporated to obtain 0.88g of a light yellow solid, with a yield of 83.2%. MS (ESI) m / z: 572.06 [M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ9.51(s,1H),7.60(s,1H),7.52(s,2H),7.41(s,1H),7.27(s,2H),6.93(s,1H),5.17(s,2H),4.10 (s,2H),3.46(s,2H),3.44(s,4H),3.08(s,4H),2.68(s,4H),1.56(d,J=16.5Hz,4H),1.43(s,2H),1.29(t,J=7.5Hz,3H).

[0247] According to the method of Example 27, the compounds of Examples 28-29 were prepared.

[0248] Example 28: 2,3-dichlorobenzyl (1-ethyl-5-(4-(2-(3-hydroxyazetidin-1-yl)-2-oxoethyl)piperazin-1-yl)-1H-indol-3-yl)carbamate. Analytical data: MS (ESI) m / z: 560.03 [M+H] + .

[0249] Example 29: 1-(4-chlorobenzyl)-3-(1-ethyl-5-(4-(2-morpholine-2-oxoethyl)piperazin-1-yl)-1H-indol-3-yl)urea. Analytical data: MS (ESI) m / z: 539.09 [M+H] + ,537.11[MH] - .

[0250] Although the pharmacological properties of the compounds of the present invention vary with structural changes, in general, the activity possessed by the compounds of the present invention can be confirmed in vivo. The pharmacological properties of the compounds of the present invention can be confirmed by several pharmacological in vitro tests. The pharmacological tests illustrated below are carried out with the compounds of the present invention.

[0251] Study on the ATX enzyme inhibitory activity of the product of the present invention

[0252] The inhibitory effect of the indole derivatives of the present invention on ATX was tested using an ATX detection kit, with FS-3 as a substrate and Zirixetastat as a positive control.

[0253] Starting with the highest concentration of 20 μM, 10 μL of serially diluted compounds were added to each well. Glycosylated human ATX protein was used at a final concentration of 0.4 or 0.64 μg / mL and was diluted in 50 mM Tris-HCl (2-amino-2-hydroxymethyl-1,3-propanediol hydrochloride), pH 8, with 250 mM NaCl, 5 mM KCl, 1 mM MgCl, 1 mM CaCl and 0.1% fatty acid-free BSA (bovine serum albumin) to maintain a total volume of 20 μL. Compounds were added to the enzyme mixture and the resulting mixture was shaken at room temperature and incubated for 30 minutes. Fluorescence reaction was stimulated by adding 20 μL of 0.75 μM FS-3 diluted in the same buffer. After incubation at room temperature for 30 minutes (excitation wavelength 485 nm, emission wavelength 520 nm), fluorescence values ​​were read from a fluorescence excitation instrument (PerkinElmer).

[0254] Study on the antiproliferative activity of the product of the present invention

[0255] Taking RAW264.7 cells (mouse mononuclear macrophage leukemia cells) as an example, the indole derivatives of the present invention were screened for in vitro antitumor activity. The other two test cell lines were MCF-7 cells (human breast cancer cell line) and brain glioma cells U87MG (human brain astroglioma cells).

[0256] (1) After the cells have recovered and been passaged 2-3 times to stabilize, use trypsin solution (0.25%) to digest them from the bottom of the culture bottle. Pour the cell digestion solution into a centrifuge tube, and then add culture medium to terminate the digestion. Centrifuge the centrifuge tube at 800r / min for 10min, discard the supernatant and add 5mL culture medium. Mix the cells by blowing, and take 10μL of the cell suspension and add it to the cell counting plate for counting. Adjust the cell concentration to 10 4 100 μL of cell suspension was added to all wells except well A1 which was a blank well without cells. The 96-well plate was placed in an incubator and cultured for 24 h.

[0257] (2) Dissolve the test sample in 50 μL of dimethyl sulfoxide, then add an appropriate amount of culture medium to dissolve the sample into a 2 mg / mL solution, and then dilute the sample to 20, 4, 0.8, 0.16, and 0.032 μg / mL in a 24-well plate. Add 3 wells for each concentration, of which the cell growth in the two rows and two columns around is greatly affected by the environment and is only used as a blank cell well. Place the 96-well plate in an incubator and culture for 72 hours.

[0258] (3) The drug-containing culture medium in the 96-well plate was discarded, and the cells were rinsed twice with phosphate buffer solution (PBS). 100 μL of MTT (0.5 mg / mL) was added to each well and placed in the incubator for 4 hours. The MTT solution was discarded and 100 μL of dimethyl sulfoxide was added. The surviving cells were shaken on a magnetic oscillator to fully dissolve the formazan reaction product of MTT, and the result was measured in an ELISA instrument. The IC value of the drug can be calculated by the Bliss method. 50 value.

[0259] The results of the ATX enzyme inhibitory activity of the compounds are shown in Table 5 (unit: nM).

[0260] Table 5 Results of the inhibitory activity of the compounds in the examples on ATX enzyme

[0261]

[0262] The antiproliferative activity results of the compounds on RAW264.7 cells, MCF-7 cells and U87MG cells are shown in Table 6 (in this table, "A" represents IC 50 Values ​​< 1 μM; "B" means 1 μM < IC 50 Values ​​<10 μM; "C" indicates IC 50 value >10 μM).

[0263] Table 6 Antiproliferative activity results of the compounds in the examples on RAW264.7 cells, MCF-7 cells and U87MG cells

[0264]

[0265]

[0266] Study on the anti-fibrotic activity of the compounds of the present invention in vivo

[0267] The Masson staining experimental method was used to investigate the inhibitory effect of compound 3 in Example 3 on pulmonary fibrosis.

[0268] (1) Modeling

[0269] The mice were anesthetized with 4% chloral hydrate and fixed in supine position. The operator held the lower lip with one hand and pulled the mouth open with forceps. The tongue of the mouse was pulled out with the other hand and pressed against the lower lip. The operator then held the mouse tracheal cannula and inserted it into the trachea through the mouth of the mouse. The drug (bleomycin hydrochloride, 4U / kg) in the syringe was slowly pushed out. The tracheal cannula was then carefully pulled out.

[0270] (2) Administration

[0271] Preparation of drug solution: First, add the drug to 1% DMSO and 50% PEG 400 and ultrasonicate for 5 minutes, then add 5% Tween 80 and 44% normal saline and ultrasonicate for 5 minutes. The drug-containing preparation finally becomes clear and transparent.

[0272] KM mice were randomly divided into 3 groups according to the random number table method: control group (Control), bleomycin group (Model), bleomycin + compound 3 (3) group, 12 mice in each group (the latter two groups were modeled with bleomycin according to the pre-experimental dose), and samples were collected on the 28th day. The mice were weighed once a day to adjust the dosage. The test group was gavaged once a day (compound 3, 40 mg / kg), and the control group and model group were gavaged with blank preparations. The drug was started from 24 hours after modeling (the first day) and stopped on the 28th day. Food but water were not allowed for 12 hours before sampling.

[0273] (3) Masson staining of lung tissue

[0274] After sampling, the lung tissue was fixed with 10% formalin, embedded in paraffin and sectioned. The collagen deposition in the lung tissue was observed by Masson staining. Figure 1 shown.

[0275] Masson staining showed that there was obvious collagen fiber deposition in the bleomycin group (Model), and compound 3 could reduce the aggregation of collagen fibers. The above results suggest that compound 3 has a good protective effect on mice with bleomycin-induced pulmonary fibrosis.

[0276] Study on the activity of the compound of the present invention in promoting hair regeneration in mice

[0277] Sixteen male Kunming mice aged 6-8 weeks were selected. After the mice were adaptively raised for 1 week, local hair on the back was removed to induce the mouse hair follicles to enter the growing period from the resting period. On the second day of depilation, mice with clean depilation and intact skin were selected and randomly divided into the Control group, Model group, 3 (30 mg / kg) and 3 (60 mg / kg) dose groups. Mice in the Model group, 3 (30 mg / kg) and 3 (60 mg / kg) dose groups were smeared with 0.2 mL of 0.1% testosterone (prepared with 50% ethanol) every day to establish a testosterone-induced pathological hair loss model; mice in the Control group were smeared with 50% ethanol. 30 minutes after the application of testosterone, drugs were given according to the groups (the experimental groups were gavagely administered compound 3 at doses of 30 mg / kg and 60 mg / kg, respectively), and the Control group and the Model group were given an equal amount of normal saline once a day for 21 consecutive days (see Figure 2 ).

[0278] Depend on Figure 2 It can be seen that the hair growth of mice on the 15th and 21st days of administration was as follows: Figure 2 The experimental results showed that the hair length of the Model group was significantly lower than that of the Control group; while the hair length of mice treated with compound 3 (30 mg / kg) and 3 (60 mg / kg) was significantly higher than that of the Model group.

[0279] From the above description, one skilled in the art can easily ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications to the invention to adapt to various usages and conditions.

[0280] No unacceptable toxic effects were observed when the compounds of the invention were administered in accordance with the present invention.

Claims

1. A 1,3,5-trisubstituted indole compound, characterized in that: The compound is a compound represented by general formula I and its enantiomer, pharmaceutically acceptable salt or prodrug: in, L is M is wherein X is O or N, Y is C or N, and Z is CH, O or N, such that X, Y, Z and the carbon and nitrogen atoms to which they are connected together form a five-membered aromatic heterocyclic ring system; Q and W are the same or different and are selected from carbonyl, (C1-C6) straight chain or branched alkyl; n is 0 or 1; R1 and R2 may be the same or different and independently selected from hydrogen, cyano, hydroxyl, (C3-C8) cycloalkyl, (C1-C6) acyl, (C1-C6) alkoxycarbonyl, (C1-C6) alkyl-substituted sulfone, (C1-C6) alkyl-substituted sulfoxide, (C6-C 10 )aryl, 4-10 membered heterocyclic group, 5-10 membered heteroaryl, aminoacyl substituted by 1-2 (C1-C6) alkyl groups, or (C1-C6) straight-chain or branched alkyl, (C2-C6) alkenyl, (C2-C6) alkynyl substituted by 1-3 identical or different substituents as follows; the substituents as follows are selected from hydrogen, halogen, hydroxyl, cyano, (C1-C3) alkoxy, carboxyl, and amino; Or R1 and R2 together with the N atom to which they are connected form a 4-10 membered heterocyclic ring containing at least one heteroatom or a 5-10 membered aromatic heterocyclic ring containing at least one heteroatom, wherein the heterocyclic ring and the aromatic heterocyclic ring may be substituted by 1-3 identical or different following groups: halogen, cyano, nitro, hydroxyl, mercapto, carboxyl, (carbon)-oxo, (sulfur)-oxo, (sulfur)-dioxo, (C1-C6)alkyl, methoxy, hydroxy-substituted (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)acyl, (C2-C6)acyloxy, (C3-C8)cycloalkyl, (C1-C6)alkoxycarbonyl; R3 is unsubstituted or substituted with at least one of the following substituents: (C1-C6) alkyl, (C2-C6) alkenyl, (C2-C6) alkynyl, (C3-C8) cycloalkyl, (C6-C 10 ) aryl, 5- to 10-membered heteroaryl or 4- to 10-membered heterocyclic group, the following substituents are hydrogen, halogen, hydroxyl, cyano, methoxy, carboxyl, amino; R4 is C6~C 10 Aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclic group, unsubstituted or substituted by 1-3 substituents which may be the same or different from the following (C1-C6) alkyl, the following substituents are selected from hydrogen, halogen, C1-C3 alkoxy, C1-C3 alkylthio, cyano, hydroxyl; the aryl, heteroaryl or heterocyclic group may also be optionally substituted independently by 1-4 of the following groups which may be the same or different: halogen, cyano, nitro, hydroxyl, thiol, carboxyl, (C1-C6) alkoxy, (C1-C6) acyl, (C2-C6) acyloxy, (C1-C6) alkoxycarbonyl, (C1-C4) alkyl(oxy)sulfone.

2. The 1,3,5-trisubstituted indole compound according to claim 1, characterized in that: The compound is a compound represented by general formula I and its enantiomer, pharmaceutically acceptable salt or prodrug; Where: L is M is X is O or N, Y is C or N, and Z is CH, O or N, so that X, Y, Z and the carbon and nitrogen atoms to which they are connected together form a five-membered aromatic heterocyclic ring system; Q and W are the same or different and are selected from carbonyl, (C1-C3) straight chain or branched alkyl; n is 0 or 1; R1 and R2 may be the same or different and independently represent hydrogen, cyano, hydroxyl, (C3-C6) cycloalkyl, (C1-C4) acyl, (C1-C4) alkoxycarbonyl, (C1-C4) alkyl-substituted sulfone, (C6-C 10 ) aryl, aminoacyl substituted by 1 to 2 (C1 to C4) alkyl groups, (C1 to C4) straight or branched alkyl groups which are unsubstituted or substituted by 1 to 3 identical or different substituents as follows, wherein the substituents are independently selected from hydrogen, halogen, hydroxyl, cyano, methoxy, carboxyl, and amino; Or R1 and R2 together with the N atom to which they are attached form a 4-10 membered heterocyclic ring containing at least one heteroatom, and the heterocyclic ring may be substituted by 1-2 identical or different following groups: halogen, cyano, hydroxyl, carboxyl, (carbon)-oxo, (sulfur)-dioxo, (C1-C4)alkyl, methoxy or hydroxy-substituted (C1-C4)alkyl, (C1-C4)alkoxy, (C2-C4)acyloxy, (C1-C4)alkoxycarbonyl; R3 is unsubstituted or substituted by 1 to 2 groups independently selected from hydrogen, halogen, hydroxyl, cyano, methoxy, amino groups and the following structures: (C1-C4) straight or branched chain alkyl, (C3-C6) cycloalkyl, phenyl, 5-6 membered heteroaryl or 4-7 membered heterocyclic group; R4 is phenyl, naphthyl, 5-6 membered heteroaryl or 4-7 membered heterocyclic group, and the phenyl, naphthyl, heteroaryl or heterocyclic group may be optionally substituted independently by 1 to 3 identical or different following groups: halogen, cyano, nitro, hydroxyl, carboxyl, (C1-C4) alkoxy, (C2-C4) acyl, (C2-C4) acyloxy, (C1-C4) alkoxycarbonyl, (C1-C4) alkyl which is unsubstituted or substituted by 1 to 3 groups independently selected from hydrogen, halogen, methoxy, methylthio, cyano, hydroxyl groups.

3. The 1,3,5-trisubstituted indole compound according to claim 2, characterized in that: The compound is a compound represented by general formula I and its enantiomer, pharmaceutically acceptable salt or prodrug; Where: L is M is Q is selected from (C1-C3) alkyl; n is 0 or 1; R1 and R2 may be the same or different and may be selected from hydrogen, (C1-C4) straight or branched alkyl, (C3-C6) cycloalkyl, cyano, hydroxy, hydroxyethyl, cyanoethyl, phenyl, methoxy-substituted (C1-C4) alkyl; Or R1 and R2 together with the N atom to which they are attached form a 4-7 membered heterocyclic ring containing at least one heteroatom, and the heterocyclic ring may be substituted by 1-2 identical or different following groups: (C1-C4) straight or branched chain alkyl, (carbon)-oxo, (sulfur)-dioxo, hydroxyl, hydroxymethyl, hydroxyethyl, carboxyl, (C3-C6) cycloalkyl; R3 is (C1-C4) alkyl or (C3-C6) cycloalkyl; R4 is phenyl or pyridyl, and the phenyl or pyridyl may be optionally independently substituted by 1 to 3 identical or different following groups: halogen, cyano, hydroxyl, carboxyl, (C1-C4) alkyl, C1-C3 hydroxyalkyl, (C1-C4) haloalkyl, methoxy.

4. The 1,3,5-trisubstituted indole compound according to claim 3, characterized in that: The compound is a compound represented by general formula I and its enantiomer, pharmaceutically acceptable salt or prodrug; Where: L is M is Q is CH2 or n = 0 or 1; R1 and R2 may be the same or different and may be selected from hydrogen, hydroxyethyl, methyl, ethyl, cyclopropyl, cyclohexyl, and phenyl; Or R1 and R2 together with the nitrogen atom to which they are connected form a 4-6 membered heterocyclic ring containing at least one heteroatom; the heterocyclic ring may also be substituted by at least one C1-C3 alkyl, hydroxyl, C1-C3 hydroxyalkyl, carboxyl, (thio)-dioxo, oxo, etc., including R3 is methyl or ethyl; R4 is phenyl or pyridyl substituted as follows:

5. The 1,3,5-trisubstituted indole compound according to claim 4, characterized in that: The compound is a compound represented by general formula I and its enantiomer, pharmaceutically acceptable salt or prodrug; Where: L is M is Q is CH2 or n = 0 or 1; R1 and R2 may be the same or different and may be selected from hydrogen, hydroxyethyl, methyl, ethyl, cyclopropyl, cyclohexyl, and phenyl; Or R1 and R2 together with the nitrogen atom to which they are attached form R3 is methyl or ethyl; R4 is 6. The 1,3,5-trisubstituted indole compound according to claims 1 to 5, characterized in that: The compound is the following compound and its enantiomer, pharmaceutically acceptable salt or prodrug; 3,4,5-Trichlorobenzyl (S)-(1-ethyl-5-(4-((2-(hydroxymethyl)pyrrolidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate; 3,4,5-Trichlorobenzyl (R)-(1-ethyl-5-(4-((2-(hydroxymethyl)pyrrolidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate; 3,4,5-Trichlorobenzyl(1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate; 3,4,5-Trichlorobenzyl(1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate; 3,4-Difluorobenzyl(1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate; 2,3-Dichlorobenzyl (1-ethyl-5-(4-(anilinomethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate; 3,5-Dichlorobenzyl(1-ethyl-5-(4-((4-methylpiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate; 1-(1-ethyl-5-(4-((4-methylpiperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)-3-(4-fluorobenzyl)urea; 1-(3-chloro-4-methoxybenzyl)-3-(1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)urea; 3,5-Dichlorobenzyl(1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate; 4-(Trifluoromethyl)benzyl(1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate; 1-(4-chlorobenzyl)-3-(1-ethyl-5-(4-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)urea; 3,5-Dichlorobenzyl(1-methyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)carbamate; (R)-1-(4-methylphenyl)ethyl(5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1-methyl-1H-indol-3-yl)carbamate; 1-(4-fluorobenzyl)-3-(1-methyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)urea; N-(4-(tert-butyl)phenyl)-2-(1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-2-oxoacetamide; N-(4-(tert-butyl)phenyl)-2-(1-ethyl-5-(5-((4-hydroxypiperidin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-2-oxoacetamide; N-(4-(tert-butyl)phenyl)-2-(5-(5-((cyclohexyl)methyl)amino)methyl)-1,2,4-oxadiazol-3-yl)-1-ethyl-1H-indol-3-yl)-2-oxoacetamide; 3,5-Dimethoxybenzyl(1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate; 1-(1-ethyl-5-(5-(tetrahydropyrrol-1-ylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-3-(4-fluorobenzyl)urea; 3,5-Dichlorobenzyl(1-ethyl-5-(5-((4-hydroxypiperidin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate; 3,5-Dichlorobenzyl (S)-(1-ethyl-5-(5-((2-(hydroxymethyl)tetrahydropyrrol-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate; 3,5-Dichlorobenzyl (R)-(1-ethyl-5-(5-((2-(hydroxymethyl)tetrahydropyrrol-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate; 1-(1-ethyl-5-(5-((4-hydroxypiperidin-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)-3-(4-fluorobenzyl)urea; 1-((2,4-dichlorobenzyl)oxy)-3-(1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)urea; N-((1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamoyl)-4-methylbenzenesulfonamide; 3,5-Dichlorobenzyl(1-ethyl-5-(4-(2-oxo-2-(piperidin-1-yl)ethyl)piperazin-1-yl)-1H-indol-3-yl)carbamate; 2,3-Dichlorobenzyl(1-ethyl-5-(4-(2-(3-hydroxyazetidin-1-yl)-2-oxoethyl)piperazin-1-yl)-1H-indol-3-yl)carbamate; 1-(4-chlorobenzyl)-3-(1-ethyl-5-(4-(2-morpholin-2-oxoethyl)piperazin-1-yl)-1H-indol-3-yl)urea.

7. A pharmaceutical composition, characterized in that: The composition contains the compound represented by the general formula I of claim 1 and its enantiomer, pharmaceutically acceptable salt or prodrug.

8. An application of the 1,3,5-trisubstituted indole compound according to claim 1, characterized in that: Use of the compound represented by general formula I of claim 1 and its enantiomer, pharmaceutically acceptable salt or prodrug or the pharmaceutical composition of claim 7 in the preparation of ATX inhibitors.

9. Use of the 1,3,5-trisubstituted indole compound according to claim 1, characterized in that: Use of the compound of general formula I of claim 1 and its enantiomer, pharmaceutically acceptable salt or prodrug or the pharmaceutical composition of claim 7 in the preparation of a drug for treating or preventing tumors, fibrotic diseases and promoting hair growth in mammals.

10. The use according to claim 9, characterized in that: The tumors are mainly breast cancer or glioma, and the fibrotic diseases are mainly pulmonary fibrosis, liver fibrosis or myocardial fibrosis.

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