1,3,5-trisubstituted indole derivatives and their use

Abstract

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

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology 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 Technology

[0002] Autotaxin (ATX), a secretory glycoprotein, hydrolyzes LPC (lysophosphatidylcholine) into choline and the lipid signaling molecule LPA (lysophosphatidate). The ATX-LPA functional axis plays an important role in fibrotic and neoplastic diseases; selective inhibition of ATX can effectively control the progression of tumors and fibrosis. In recent years, research on ATX inhibitors has gradually matured. The compound Zirixetastat (WO2014202458A1) entered Phase III clinical trials in 2019 for the treatment of idiopathic pulmonary fibrosis (IPF). In addition, drugs such as BBT877 (patent number undisclosed), X-165 (WO2015154023), and PAT-409 (WO2016028686A1) are in Phase I or Phase II clinical trials.

[0003] Domestic research on ATX inhibitors mainly focuses on simple site modifications based on atomic substitution (Zirixetastat imidazopyridine structural analogs), i.e., the development of "me-too" drugs, such as five-membered and six-membered fused-ring pyrazolopyridine compounds (WO2019158107, CN202210530649.9), imidazopyridazine compounds (CN202010658085.8), and five-membered and five-membered fused-ring imidazothiazole compounds (CN202111348160.1). However, overall, there are relatively few reports on innovative structural exploration. Previously, 5-thiazolyl indole compounds (CN202010043108.4) were reported as novel ATX inhibitors (optimal compound IC50). 50 Although the value is less than 1 nM, it exhibits superior anti-fibrotic activity, but the activity needs to be further improved.

[0004] Hair loss is a significant public health issue. Numerous studies and surveys have shown that hair loss significantly impacts the quality of life and mental well-being of sufferers. Hair loss has become a major concern for many young people today, making its improvement urgent. Currently, clinical methods for improving hair loss mainly include medication and surgery, but both have significant limitations and drawbacks. Oral and topical medications (such as minoxidil and finasteride) are relatively inexpensive and can temporarily relieve hair loss symptoms, but they suffer from significant side effects, slow onset of action, unstable efficacy, and recurrence after discontinuation. Surgical treatment offers rapid results, but is expensive, and because it involves transplanting individual hair follicles and follicular units from the patient's own body, the number of follicles is limited, and the survival rate is unstable. Therefore, exploring new, safe, and effective treatments for hair loss is of paramount importance. Summary of the Invention

[0005] The purpose of this invention is 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 objectives, the present invention adopts the following technical solution:

[0007] A 1,3,5-trisubstituted indole compound, wherein the compound is represented by general formula I and its enantiomers, pharmaceutically acceptable salts, or prodrugs:

[0008]

[0009] in,

[0010] L is

[0011] M is

[0012] Where X is O or N, Y is C or N, and Z is CH, O or N, so that X, Y, Z together with the carbon and nitrogen atoms attached to them form a five-membered aromatic heterocyclic system;

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

[0014] n is 0 or 1;

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

[0016] Alternatively, R1 and R2 together with the N atom to which they are attached form a 4- to 10-membered heterocycle containing at least one heteroatom or a 5- to 10-membered aromatic heterocycle containing at least one heteroatom, wherein the heterocycle and aromatic heterocycle may be substituted by 1 to 3 identical or different of the following groups: halogen, cyano, nitro, hydroxyl, mercapto, carboxyl, (carbon)-oxo, (sulfo)-oxo, (sulfo)-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 an unsubstituted (C1-C6) alkyl, (C2-C6) alkenyl, (C2-C6) ynyl, (C3-C8) cycloalkyl, (C6-C6) cycloalkyl, or (C3-C8) alkyl group, substituted with at least one of the following substituents. 10 ) aryl, 5-10 membered heteroaryl or 4-10 membered heterocyclic group, wherein the following substituents are hydrogen, halogen, hydroxyl, cyano, methoxy, carboxyl or amino;

[0018] R4 is C6~C 10 The aryl, 5-10-membered heteroaryl, 4-10-membered heterocyclic, unsubstituted or (C1-C6) alkyl group substituted with 1-3 identical or different substituents selected from hydrogen, halogen, C1-C3 alkoxy, C1-C3 alkylthio, cyano, and hydroxyl groups may also optionally be independently substituted with 1-4 identical or different substituents: halogen, cyano, nitro, hydroxyl, mercapto, carboxyl, (C1-C6)alkoxy, (C1-C6) acyl, (C2-C6) acyloxy, (C1-C6) alkoxycarbonyl, and (C1-C4) alkyl(sulfone) group.

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

[0020] In the formula:

[0021] L is

[0022] M is

[0023] Where X is O or N, Y is C or N, and Z is CH, O or N, so that X, Y, Z together with the carbon and nitrogen atoms attached to them form a five-membered aromatic heterocyclic system;

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

[0025] n is 0 or 1;

[0026] R1 and R2 may be the same or different and independently composed of hydrogen, cyano, hydroxyl, (C3-C6)cycloalkyl, (C1-C4) acyl, (C1-C4) alkoxycarbonyl, (C1-C4) alkyl-substituted sulfone, (C6-C4) alkyl-substituted sulfone, (C6-C4) cycloalkyl, ... 10 ) aryl, aminoacyl group substituted with 1 to 2 (C1 to C4) alkyl groups, unsubstituted or (C1 to C4) straight-chain or branched alkyl groups substituted with 1 to 3 identical or different substituents selected independently from hydrogen, halogen, hydroxyl, cyano, methoxy, carboxyl, and amino.

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

[0028] R3 is an unsubstituted or substituted structure consisting of one or two independently selected groups of hydrogen, halogen, hydroxyl, cyano, methoxy, or amino groups: (C1-C4) straight-chain or branched 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, wherein the phenyl, naphthyl, heteroaryl, or heterocyclic group may optionally be independently substituted by 1-3 identical or different of the following groups: halogen, cyano, nitro, hydroxyl, carboxyl, (C1-C4)alkoxy, (C2-C4) acyl, (C2-C4) acyloxy, (C1-C4) alkoxycarbonyl, unsubstituted or (C1-C4) alkyl groups independently selected from hydrogen, halogen, methoxy, methylthio, cyano, or hydroxyl groups;

[0030] More preferably, the compound is a compound of general formula I and its enantiomers, a pharmaceutically acceptable salt or a prodrug;

[0031] In the formula:

[0032] L is

[0033] M is

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

[0035] n is 0 or 1;

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

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

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

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

[0040] Further preferably, the compound is a compound of general formula I and its enantiomers, a pharmaceutically acceptable salt or a prodrug;

[0041] In the formula:

[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 are selected from hydrogen, hydroxyethyl, methyl, ethyl, cyclopropyl, cyclohexyl, and phenyl.

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

[0047] R3 represents methyl or ethyl;

[0048] R4 is a phenyl or pyridyl group substituted with the following:

[0049] More preferably, the compound is a compound of general formula I and its enantiomers, a pharmaceutically acceptable salt or a prodrug;

[0050] In the formula:

[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 are selected from hydrogen, hydroxyethyl, methyl, ethyl, cyclopropyl, cyclohexyl, and phenyl.

[0055] Alternatively, R1 and R2 together with the nitrogen atoms they are attached to form...

[0056] R3 represents methyl or ethyl;

[0057] R4 is

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

[0059] 3,4,5-Trichlorobenzyl(S)-(1-ethyl-5-(4-((2-(hydroxymethyl)pyrrolidone-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)pyrrolidone-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-indole-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-(tetrahydropyrrolo-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)tetrahydropyrrolo-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)tetrahydropyrrolo-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-hydroxyazacyclobutane-1-yl)-2-oxoethyl)piperazin-1-yl)-1H-indole-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] This invention includes a method for preparing compounds of general formula I and intermediates thereof.

[0089] Furthermore, according to some common methods in the field to which this invention pertains, the indole derivatives of general formula I in this invention can react with acids to form pharmaceutically acceptable salts. Pharmaceutically acceptable addition salts include addition salts of inorganic and organic acids, with salts formed with the following acids being 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 react with bases to form pharmaceutically acceptable salts, with salts formed with the following bases being 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 salting the piperazine structure in Compound 8 with hydrochloric acid.

[0090] Furthermore, this invention also includes prodrugs derived from the present invention. These prodrugs are indole derivatives of general formula I, which may possess weak or even no activity on their own, but are converted into their corresponding biologically active forms under physiological conditions (e.g., through metabolism, solvent degradation, or other means) after administration. The acetyl-protected derivatives are the prodrugs of their corresponding hydrolysis products, the indole derivatives (the products corresponding to Examples 1-3, 10-14, and 21-24 are their acetyl-protected prodrugs).

[0091] General synthesis steps

[0092] The examples and preparation methods provided below further illustrate and demonstrate the general formula compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparation methods is not intended to limit the scope of the invention in any way, but merely to provide methods for practicing the invention.

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

[0094] The proton NMR spectra of the compounds were determined using a Bruker ARX-400 / 600, and the mass spectra were determined using an Agilent 1100 LC / MS. All reagents used were of analytical or chemical purity.

[0095] The compounds of this 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 Method G

[0109]

[0110] A pharmaceutical composition comprising a compound of formula I and its enantiomers, a pharmaceutically acceptable salt, or a prodrug.

[0111] The use of the 1,3,5-trisubstituted indole compound, the use of the compound of general formula I and its enantiomers, pharmaceutically acceptable salts or prodrugs or the pharmaceutical composition thereof in the preparation of an ATX inhibitor.

[0112] The use of the 1,3,5-trisubstituted indole compound of general formula I, its enantiomers, pharmaceutically acceptable salts or prodrugs, or the pharmaceutical composition thereof, in the preparation of a medicament for the treatment or prevention of tumors, fibrotic diseases, and the promotion of hair growth in mammals.

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

[0114] In vitro enzyme activity tests have shown that the compounds of this invention possess strong in vitro ATX inhibitory activity. Therefore, they can be used to prepare drugs for the treatment and / or prevention of 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 nodes, soft tissue, head and neck, thyroid cancer, leukemia, neuroblastoma, etc.; and fibrotic diseases mainly including pulmonary fibrosis, liver fibrosis, myocardial fibrosis, kidney fibrosis, etc.

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

[0116] Preliminary in vivo experiments have shown that the preferred compounds in this invention have a good prognostic effect on bleomycin-mediated pulmonary fibrosis tissue, demonstrating the great potential of this series of compounds as therapeutic drugs for pulmonary fibrosis.

[0117] The active compounds of the present invention, or pharmaceutically acceptable salts thereof, can be used alone as the sole antitumor or antifibrotic agent, or in combination with existing antitumor / antifibrotic agents (such as pirfenidone, nintedanib, paclitaxel, cisplatin, etc.). Combination therapy is achieved by administering the various therapeutic components simultaneously, sequentially, or separately.

[0118] Preliminary pharmacological experiments have shown that the preferred compounds in this invention have a significant promoting effect on hair regeneration in mice, demonstrating the great potential of this series of compounds as a treatment for hair loss.

[0119] This invention designs and synthesizes a series of 1,3,5-trisubstituted indole derivatives, all of which are novel structural types, exhibiting significant structural novelty. Computational studies show that the compounds of this invention employ an allosteric binding mode. In vitro and in vivo activity screening indicates that these compounds possess significant ATX enzyme inhibitory activity, and demonstrate excellent antitumor and fibrosis inhibitory activities. Their in vitro and in vivo activities are superior to the positive control drugs Zirixetastat and PF-8380, providing a promising alternative candidate molecule for the treatment of tumors and fibrotic diseases, demonstrating significant inventiveness and good application prospects. Furthermore, the experiments revealed that the compounds of this invention significantly promote hair growth in mice, indicating significant potential for further exploration, development, and application. Attached Figure Description

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

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

[0122] The following examples further illustrate the compounds, pharmaceutical compositions, and their applications provided by the present invention.

[0123] The substituents of the compounds prepared in Examples 1 to 29 of this invention are shown below:

[0124]

[0125] In the general formula compound structure, M is Q is methylene, n=1, and R3 is ethyl. The specific compounds in Examples 1-12 are described in Table 1.

[0126] Table 1 Examples 1-12

[0127]

[0128] In the general formula compound structure, M is n = 1, R3 is a methyl group. The specific compounds in Examples 13-15 are described in Table 2.

[0129] Table 2 Examples 13-15

[0130]

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

[0132] Table 3 Examples 16-26

[0133]

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

[0135] Table 4 Examples 27-29

[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, 12 g (0.3 mol) of 60% NaH was added to 100 mL of dry DMF and stirred. At 0 °C, 80 mL of 20 g (0.124 mol) DMF solution was added dropwise. After H2 escaped, 15 mL (0.185 mol) of iodoethane was added, and the reaction was allowed to proceed at room temperature for 2 h. After the reaction was complete, 600 mL of saturated NH4Cl solution was added to a 2 L beaker and stirred. The reaction solution was then slowly poured into the beaker to quench the reaction. After stirring for 20 min, the mixture was filtered, and the filter cake was dried to obtain 22.8 g of a 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 and stirred. Then, (CF3CO)2O (20.79 g, 0.099 mol) was added dropwise at 0 °C. After the addition was complete, the reaction was allowed to proceed at room temperature for 2.5 h. After the reaction was complete, 300 mL of water was added to the reaction flask, the mixture was stirred, filtered, and the filter cake was dried to give 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 5 M NaOH and refluxed for 2 h. After the reaction was complete, 120 mL of water was added to the reaction flask, stirred, and the pH was adjusted to 3 with HCl. A solid precipitated, was filtered, and the filter cake was dried to give 11.5 g of product, with a yield of 85.0%.

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

[0145] At room temperature, 10.8 g (0.046 mol) of 1-ethyl-5-nitro-1H-indole-3-carboxylic acid was added to dry toluene. 6.5 mL of TEA and 7.5 mL of DPPA were added with stirring. After 30 min, 12.25 g (0.069 mol) of 3,5-dichlorobenzyl alcohol was added, and the mixture was refluxed for 2 h. After the reaction was complete, the solvent was evaporated, water was added, and a solid precipitated. The filtrate was discarded after monitoring for UV absorption. The filter cake was transferred to a beaker, DCM was added, and stirred. Insoluble matter was present. The mixture was filtered, and the filter cake was dried to give 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-indole-3-yl)carbamate

[0147] At room temperature, 8.0 g (0.02 mol) of 3,5-dichlorobenzyl (1-ethyl-5-nitro-1H-indol-3-yl) carbamate was dissolved in 30 mL of MeOH, SnCl₂·2H₂O (5 eq) was added, followed by 10 mL of concentrated hydrochloric acid, and the mixture was refluxed for 3 h. After the reaction was complete, 60 mL of water was added to the reaction flask, the mixture was stirred, and the pH was adjusted to 8 with saturated Na₂CO₃ solution. The mixture was 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-indole-3-yl)carbamate

[0149] At 0°C, 3,5-dichlorobenzyl (5-amino-1-ethyl-1H-indol-3-yl)carbamate (3 g, 0.006 mol) was added to 10 mL of H₂O and stirred. 4.5 mL of concentrated HCl was added, followed by dropwise addition of 1 M NaNO₂ aqueous solution. After 0.5 h, 1 M NaN₃ aqueous solution was added dropwise, and the reaction was allowed to proceed at room temperature for 1.5 h. After the reaction was complete, 30 mL of water was added, and the mixture was extracted with DCM. The organic layer was evaporated to dryness, and hexane was added and stirred. The mixture was filtered, and the filter cake was dried to obtain 1.60 g 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-indole-3-yl)carbamate

[0151] At 0°C, 1.5 g (0.004 mol) of 3,5-dichlorobenzyl (5-azido-1-ethyl-1H-indol-3-yl) carbamate was dissolved in DMF and stirred. Propylene alcohol (0.25 g, 0.0045 mol), VcNa (0.15 g, 0.0009 mol), CuI (0.35 g, 0.002 mol), and DIPEA (0.75 g, 0.0074 mol) were added sequentially. After 10 min, the reaction was continued at room temperature for 3 h. After the reaction was complete, 6 mL of ammonia was added, and the mixture was stirred. Ice water was added, and the mixture was filtered. The filter cake was washed with methanol, filtered again, 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-indole-3-yl)carbamate

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

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

[0155] At room temperature, 0.1 g of 3,4,5-trichlorobenzyl(5-(4-(chloromethyl)-1H-1,2,3-triazol-1-yl)-1-ethyl-1H-indol-3-yl)carbamate was dissolved in 4 mL of MeCN and stirred. L-prolyl (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 was complete, 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 the crude product. The target compound was purified by column chromatography in 20% yield. 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] Following the method of Example 1, the compounds of Examples 2-4 were obtained by N-alkylation reaction of intermediate a9 with a small molecule amine.

[0157] Example 2: 3,4,5-Trichlorobenzyl(R)-(1-ethyl-5-(4-((2-(hydroxymethyl)pyrrolidone-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 (1 g, 0.0035 mol) was dissolved in THF (8 mL), stirred, and zinc powder (1.36 g) was added. Saturated ammonium chloride solution (8 mL) was then added dropwise, and the mixture was refluxed for 3 h. After the reaction was complete, the mixture was filtered through a diatomaceous earth filter. The filter cake was washed with EA (30 mL) until the yellow color disappeared. The filtrate was transferred to a separatory funnel for separation. The organic layer was washed with saturated brine and evaporated to dryness to obtain 0.75 g of a 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, 0.72 g (0.0028 mol) of 1-(5-amino-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one was added to 8 mL of H₂O, followed by 2.2 mL of 37% HCl. Then, 1 M NaNO₂ aqueous solution was added dropwise. After the addition was complete, the mixture was stirred for 0.5 h, followed by the addition of 1 M NaN₃ aqueous solution. The reaction was then allowed to proceed at room temperature for 2 h. After the reaction was complete, 15 mL of water was added, and the mixture was stirred for 10 min. The mixture was then 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, 0.52 g (0.02 mol) of 1-(5-azido-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroethane-1-one was dissolved in 5 mL of DMF and stirred. Propynol (1.2 eq), VcNa (0.2 eq), CuI (0.5 eq), and DIPEA (2 eq) were added sequentially. After 10 min, the reaction was allowed to proceed at room temperature for 3 h. Upon completion of the reaction, 4 mL of 5% ammonia solution was added, stirred, and then ice water was added. The mixture was filtered, and 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, 0.54 g (0.0016 mol) of 1-(1-ethyl-5-(4-(hydroxymethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one was dissolved in 6 mL of DCM, thionyl chloride was added dropwise, and 1 drop of DMF was added as a catalyst. The reaction was carried out at room temperature for 2 h. After the reaction was complete, 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 give 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, 0.29 g (0.0008 mol) of 1-(5-(4-(chloromethyl)-1H-1,2,3-triazol-1-yl)-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one was dissolved in acetonitrile (4 mL), stirred, and then potassium carbonate (5 eq) and morpholine (2 eq) were added. The reaction was carried out at room temperature for 2 h. After the reaction was complete, 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 give 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, 0.2 g of 1-(1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)-2,2,2-trifluoroethyl-1-one was added to a reaction flask, followed by 6M sodium hydroxide solution. The mixture was heated to reflux for 2 h. After the reaction was complete, the pH was adjusted to 3, and a black solid precipitated. The solid was filtered, and the solid was identified as the product. Product residue remained in the liquid. Extraction was performed using DCM, and a white solid precipitated in the aqueous layer. The solid was filtered, and the solid was identified as 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, 0.1 g of 1-ethyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indole-3-carboxylic acid was dissolved in dry toluene and stirred. TEA (2 eq) and DPPA (1.5 eq) were added, and after 30 min, 1.5 eq of 3,4-difluorobenzyl alcohol was added. The mixture was heated to reflux for 2 h. After the reaction was complete, the solvent was evaporated, and the mixture was separated by adding water and dichlorophenate. The organic layer was washed with saturated brine and evaporated to dryness. The product was purified by column chromatography in 24.0% yield. 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] Following the method of Example 5, intermediate b6 was used as a starting material to undergo a 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-indole-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] Intermediate c1 of the C-series compounds was prepared using a method similar to that used in 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, 2 g of 1-ethyl-5-(4-((4-hydroxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1H-indole-3-carboxylic acid, 2.2 eq of acetic anhydride, 2.2 eq of TEA, and 0.1 eq of DMAP were added sequentially to 15 mL of DCM, and the mixture was reacted at room temperature for 2 h. After the reaction was complete, water was added and the mixture was separated from the DCM. The organic layer was evaporated to dryness to give 1.22 g of a 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, 0.2 g of 5-(4-((4-acetoxypiperidin-1-yl)methyl)-1H-1,2,3-triazol-1-yl)-1-ethyl-1H-indole-3-carboxylic acid was dissolved in dry toluene and stirred. TEA (2 eq) and DPPA (1.5 eq) were added, and after 30 min, 1.5 eq of 3,4-difluorobenzyl alcohol was added. The mixture was then heated to reflux for 2 h. After the reaction was complete, the solvent was evaporated, and the mixture was separated from the solid by adding water and dichlorophenoxyacetic acid. The organic layer was washed with saturated brine and evaporated to dryness to give 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, 0.1 g 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 was dissolved in 4 mL of dioxane, and 1.5 M NaOH solution was added with stirring. The reaction was carried out at 40 °C. After the reaction was complete, water was added, and the mixture was separated from DCM. The organic layer was washed with saturated brine and evaporated to dryness to obtain the crude product. The crude product was purified by column chromatography in 23% yield. 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] The compounds of Examples 11-14 were prepared according to the method of Example 10.

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

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

[0199] 1.42 g of 2-(5-cyano-1-ethyl-1H-indol-3-yl)-2-oxoacetyl chloride was dissolved in 15 mL of dry DCM. 2 eq of DIPEA was added, followed by slow dropwise addition of 1.5 eq of 4-(tert-butyl)aniline dissolved in dry DCM. After the addition was complete, the mixture was stirred at room temperature for 2 h, and the reaction was monitored by TLC. After the reaction was complete, the organic phase was first washed with 0.5 M HCl to separate it, then washed with saturated NaHCO3, and the organic layer was evaporated to dryness to give 1.83 g of a 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, and hydroxylamine hydrochloride (1.5 eq) and TEA (2 eq) were added sequentially. The mixture was heated to 70 °C and reacted overnight, monitored by TLC. After the reaction was complete, part of the solvent was evaporated, and the mixture was cooled in cold hydrazine, where a solid precipitated. Filtering yielded 1.02 g of a white solid, with a yield of 52.3%.

[0202] Step 4: Synthesis of N-(4-(tert-butyl)phenyl)-2-(5-(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, and chloroacetyl chloride (1.2 eq) was added dropwise. The mixture was heated to 100 °C and reacted for 4 h, monitored by TLC. After the reaction was complete, water was added and stirred, and the mixture was filtered to obtain 0.95 g of a brownish-yellow solid, 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-(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, and diethylamine (1.2 eq) was added. The mixture was refluxed for 12 h, and the reaction was monitored by TLC. After the reaction was completed, water was added and stirred. The mixture was extracted with DCM, evaporated to dryness to obtain the crude product, and purified by column chromatography to obtain the 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] The compounds of Examples 17-18 were prepared according to the method of Example 16.

[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-oxoacetamide. 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-oxoacetamide. 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 of DMF. Trifluoroacetic anhydride (2 eq) was added dropwise under ice bath conditions. After the addition was complete, the reaction was allowed to proceed at room temperature for 3 h. Upon completion of the reaction, a solid precipitated out. After filtration and drying, 6.12 g of a pure white solid was obtained, 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), and TEA (2 eq) were successively dissolved in ethanol and refluxed for 4 h. After the reaction was completed, 20 mL of water was added, and the ethanol was evaporated to remove it, resulting in the precipitation of a large amount of white solid. The solid was filtered, and the pH of the filtrate was adjusted to 5.5–6, resulting in the precipitation of more solid. The filter cakes were combined to obtain 3.2 g of solid, with a yield of 85.1%.

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

[0215] 3 g of 1-ethyl-N'-hydroxy-3-(2,2,2-trifluoroacetyl)-1H-indole-5-carboximide was added to a 30% sodium hydroxide solution and refluxed for 3 h. After the reaction was complete, the solution was cooled to room temperature, and the pH was adjusted to 4.5–5, resulting in the precipitation of a grayish-white solid. After filtration and drying, 2.05 g of filter cake was obtained, 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] 1.5 g of 5-(5-(chloromethyl)-1,2,4-oxadiazol-3-yl)-1-ethyl-1H-indole-3-carboxylic acid and 1.2 eq of chloroacetyl chloride were dissolved in 15 mL of DMF and reacted at 90 °C for 2 h. After the reaction was complete, the solution was cooled to room temperature, water was added, and a light gray solid precipitated. After filtration and drying, 1.38 g of a grayish-white solid was obtained, 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] 0.5 g of 5-(5-(chloromethyl)-1,2,4-oxadiazol-3-yl)-1-ethyl-1H-indole-3-carboxylic acid, 1.2 eq of morpholine, and 2 eq of potassium carbonate were added to acetonitrile and refluxed for 30 min. After the reaction was complete, the solvent was evaporated, water was added, and the pH was adjusted to 4.5-5. 0.42 g of a white solid 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] 0.1 g of 1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indole-3-carboxylic acid was added to a mixture of TEA (2 eq) and DPPA (2 eq) in dried toluene, and the reaction was carried out at room temperature for 30 min. Then, 1.2 eq of 3,5-dimethoxybenzyl alcohol was added, and the mixture was refluxed for 1 h. After the reaction was complete, toluene was removed by vacuum distillation, water was added, and the mixture was extracted with DCM. The crude product was evaporated to dryness and purified by column chromatography to obtain a pure product in 18% yield. 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] The compounds of Examples 20-24 were prepared according to the method of Example 19.

[0223] Example 20: 1-(1-ethyl-5-(5-(tetrahydropyrrolo-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)tetrahydropyrrolo-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)tetrahydropyrrolo-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 °C; 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] 0.1 g of 1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indole-3-carboxylic acid was added to a mixture of TEA (2 eq) and DPPA (2 eq) in dried toluene, and the reaction was carried out at room temperature for 30 min. Then, 1.2 eq of 2,4-dichlorobenzylhydroxylamine was added, and the mixture was refluxed for 1 h. After the reaction was complete, toluene was removed by vacuum distillation, water was added, and the mixture was extracted with DCM. The crude product was purified by column chromatography to obtain a pure product in 33.2% yield. 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] 0.1 g of 1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indole-3-carboxylic acid was added to a mixture of TEA (2 eq) and DPPA (2 eq) in dried toluene, and the reaction was carried out at room temperature for 30 min. Then, 1.2 eq of 4-methylbenzenesulfonamide was added, and the mixture was refluxed for 1 h. After the reaction was complete, toluene was removed by vacuum distillation, water was added, and the mixture was extracted with DCM. The crude product was purified by column chromatography to obtain a pure product in 27.1% yield. 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, 50.1 g (2.5 mol) of 60% NaH was added to 500 mL of DMF, stirred, and slowly heated to 40 °C. 194.97 g (1.0 mol) of 5-bromoindole was added. After H₂ escaped, the mixture was cooled to 25 °C, and 234.0 g (1.5 mol) of iodoethane was added. The reaction was allowed to proceed at room temperature for 2 h. After the reaction was complete, 800 mL of water was added, and the mixture was stirred for 30 min. The mixture was then filtered to obtain 195.1 g 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, 89.2 g (0.4 mol) of intermediate 1-(5-bromo-1-ethyl-1H-indol-3-yl)-2,2,2-trifluoroacetyl-1-one (g2) was dissolved in 400 mL of DMF. The mixture was stirred, and 83 mL (0.6 mol) of trifluoroacetic anhydride was added dropwise under ice bath conditions. After the addition was complete, the reaction was allowed to proceed at room temperature for 1.5 h. After the reaction was complete, the solution was poured into 2500 mL of water, stirred for 30 min, and then filtered to obtain 98.5 g of solid, with a yield of 78.4%.

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

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

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

[0240] At room temperature, 67.6 g (0.25 mol) of intermediate G4 was dissolved in 300 mL of dry toluene. While stirring, 34.6 g of N-Boc piperazine (0.37 mol), 48.2 g of sodium tert-butoxide (0.5 mol), and 7.5 g of JohnPhos ligand (0.025 mol) were added sequentially to the solution. Finally, 10.98 g of Pd2(dba)3 (0.012 mol) was added. Under N2 protection, the mixture was heated to 115 °C for 2 h. After the reaction was completed by TLC monitoring, the mixture was filtered through a diatomaceous earth filter. The diatomaceous earth was washed with ethyl acetate, and the filtrate was evaporated to dryness. 300 mL of n-hexane was added, and the mixture was stirred for 30 min. The mixture was then filtered, and the filter cake was washed with n-hexane (3 × 60 mL) and discarded. The n-hexane layer was evaporated to dryness to obtain 73.3 g of solid, with a yield of 85.6%.

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

[0242] In a 1000 mL round-bottom flask, 56 g (0.15 mol) of intermediate g5 was dissolved in 300 mL of dry toluene. Then, 45.5 g (0.45 mol) of triethylamine and 49.5 g (0.18 mol) of DPPA were added to the solution. The mixture was stirred at 80 °C for 30 min, followed by the addition of 31.9 g (0.18 mol) of 3,5-dichlorobenzyl alcohol. The mixture was refluxed and stirred for 2 h. After the reaction was completed by TLC monitoring, the toluene was evaporated to dryness, 500 mL of water was added, and the mixture was stirred and filtered. The liquid layer was discarded, and the upper solid layer was added to 300 mL of acetonitrile. The mixture was stirred, filtered, and the filter cake was washed with acetonitrile (3 × 100 mL). After drying, 54.2 g of solid was obtained, 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, 120 mL of HCl solution (4 M) containing dioxane was added to a 500 mL round-bottom flask. 10.9 g (0.02 mol) of intermediate g6 was slowly added in portions with stirring, and the reaction was stirred for 2 h at room temperature. After the reaction was completed by TLC monitoring, dioxane was removed under reduced pressure. 500 mL of water was slowly added with stirring, and the mixture was extracted with 200 mL of dichloromethane. The organic layer was discarded, and the aqueous layer was adjusted to pH 10 with 2N sodium hydroxide solution and extracted with dichloromethane (3 × 150 mL). The organic layers were combined, washed successively with saturated brine and water, dried over anhydrous sodium sulfate, and the dichloromethane was evaporated to dryness to obtain 8.5 g 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 50 mL pear-shaped flask, 0.89 g g7 (0.002 mol) and 0.85 g chloroacetylpiperidine (0.005 mol) were dissolved in 8 mL acetonitrile. 0.69 g potassium carbonate (0.005 mol) was added with stirring, and the mixture was refluxed for 3 h with stirring. After the reaction was completed by TLC monitoring, the potassium carbonate was removed by filtration, and the acetonitrile was removed by vacuum distillation. The residue was dissolved in 20 mL of dichloromethane and extracted with 4M hydrochloric acid (2 x 15 mL). The organic layer was discarded, and the aqueous layer was adjusted to pH 10 with 6N sodium hydroxide solution and extracted with dichloromethane (3 x 10 mL). The organic layers were combined, washed successively with saturated brine and water, dried over anhydrous sodium sulfate, and the dichloromethane was evaporated to dryness to give 0.88 g of a pale 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] The compounds of Examples 28-29 were prepared according to the method of Example 27.

[0248] Example 28: 2,3-Dichlorobenzyl (1-ethyl-5-(4-(2-(3-hydroxyazacyclobutane-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-morpholin-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 this invention vary with structural variations, generally speaking, the activities of the compounds of this invention can be demonstrated in vivo. The pharmacological characteristics of the compounds of this invention can be confirmed through several in vitro pharmacological tests. The pharmacological tests illustrated below were performed using the compounds of this invention.

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

[0252] Using FS-3 as a substrate and Zirixetastat as a positive control, the inhibitory effect of the indole derivative of the general formula of this invention on ATX was tested using an ATX detection kit.

[0253] Starting with a maximum concentration of 20 μmol, add 10 μL of serially diluted compound to each well. The final concentration used for glycosylated human ATX protein was 0.4 or 0.64 μg / mL, diluted in 50 mM Tris-HCl (2-amino-2-hydroxymethyl-1,3-propanediol hydrochloride) at pH 8, with 250 mmol of sodium chloride, 5 mmol of potassium chloride, 1 mmol of magnesium chloride, 1 mmol of calcium chloride, and 0.1% fatty acid-free BSA (bovine serum albumin) to a total volume of 20 μL. Add the compound to the enzyme mixture, shake the resulting mixture at room temperature, and incubate for 30 min. Add 20 μL of 0.75 μM FS-3 diluted in the same buffer to excite the fluorescence reaction. After incubation at room temperature for 30 min (excitation wavelength 485 nm, emission wavelength 520 nm), read the fluorescence value from a PerkinElmer fluorescence excitation device.

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

[0255] Using RAW264.7 cells (mouse mononuclear macrophage leukemia cells) as an example, the in vitro antitumor activity of the indole derivative of the general formula of this invention was screened. The other two test cell lines were MCF-7 cells (human breast cancer cell line) and U87MG glioma cells (human glioblastoma cells).

[0256] (1) After cell resuscitation and stabilization through 2-3 passages, digest the cells from the bottom of the culture flask using trypsin solution (0.25%). Pour the cell digestion solution into a centrifuge tube, followed by the addition of culture medium to terminate the digestion. Centrifuge the tube at 800 rpm for 10 min, discard the supernatant, add 5 mL of culture medium, mix the cells by pipetting, and add 10 μL of the cell suspension to a cell counting chamber for counting. Adjust the cell concentration to 102. 4 Cells / well. Except for well A1, which was a blank well with no cells, 100 μL of cell suspension was added to all other wells of the 96-well plate. The 96-well plate was then incubated in an incubator for 24 hours.

[0257] (2) Dissolve the test sample in 50 μL of dimethyl sulfoxide, then add an appropriate amount of culture medium to dissolve the sample to a concentration of 2 mg / mL. Dilute the sample to 20, 4, 0.8, 0.16, and 0.032 μg / mL in a 24-well plate. Add each concentration to three wells. The two outer rows and two columns of cells are more susceptible to environmental influences and are used only as blank wells. Incubate the 96-well plate in an incubator for 72 h.

[0258] (3) Discard the drug-containing culture medium in the 96-well plate, wash the cells twice with phosphate-buffered saline (PBS), add 100 μL of MTT (0.5 mg / mL) to each well, incubate for 4 hours, then discard the MTT solution and add 100 μL of dimethyl sulfoxide. Shake on a magnetic oscillator to fully dissolve the surviving cells and the MTT reaction product formazan, then measure the results using a microplate reader. The IC50 of the drug can be determined using the Bliss method. 50 value.

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

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

[0261]

[0262] The antiproliferative activity of the compounds against RAW264.7, MCF-7, and U87MG cells is shown in Table 6 (in this table, "A" indicates IC50). 50 Value < 1 μM; "B" indicates 1 μM < IC 50 Value <10μM; "C" indicates IC 50 Value > 10 μM).

[0263] Table 6 shows the antiproliferative activity of the compounds in the examples against RAW264.7, MCF-7, and U87MG cells.

[0264]

[0265]

[0266] In vivo antifibrotic activity study of the compound of this invention

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

[0268] (1) Modeling

[0269] Mice were anesthetized with 4% chloral hydrate, fixed in a supine position, and the operator used one hand to hold the lower lip with forceps and pull the mouth open, while the other hand used forceps to pull out the mouse's tongue and place it close to the lower lip. Then the operator held the endotracheal tube and inserted it into the trachea through the mouse's mouth, slowly pushing the drug (bleomycin hydrochloride, 4 U / kg) into the syringe, and then carefully removing the endotracheal tube.

[0270] (2) Administration

[0271] Drug solution preparation: First, add the drug to 1% DMSO and 50% PEG 400 and sonicate for 5 minutes. Then add 5% Tween 80 and 44% physiological saline and sonicate for 5 minutes. The final drug-containing preparation is clear and transparent.

[0272] KM mice were randomly divided into three groups using a random number table: control group, bleomycin drip group, and bleomycin drip + compound 3 (3) group, with 12 mice in each group (the latter two groups were modeled using bleomycin based on the pre-experimental dose). Samples were collected on day 28. The weight of the mice was measured once a day to adjust the dosage. The test group was administered compound 3 by gavage once a day (40 mg / kg), while the control group and model group were administered a blank preparation by gavage. Drug administration began 24 hours after modeling (day 1) and continued until day 28. The mice were fasted for 12 hours before sample collection, but water was allowed.

[0273] (3) Masson staining of lung tissue

[0274] After sampling, lung tissue was fixed in 10% formalin, paraffin-embedded, and sectioned. Masson staining was used to observe collagen deposition in the lung tissue. The results are as follows: Figure 1 As shown.

[0275] Masson staining revealed significant collagen fiber deposition in the bleomycin group (Model), and compound 3 reduced collagen fiber aggregation. These results suggest that compound 3 has a good protective effect against bleomycin-induced pulmonary fibrosis in mice.

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

[0277] Sixteen male Kunming mice aged 6-8 weeks were selected. After one week of acclimatization, the hair on the back of the mice was removed to induce the hair follicles to enter the anagen phase from the quiescent phase. On the second day after hair removal, mice with clean hair removal and no skin damage were randomly divided into Control group, Model group, and three (30 mg / kg) and three (60 mg / kg) dose groups. Mice in the Model group, three (30 mg / kg) and three (60 mg / kg) dose groups were treated with 0.2 mL of 0.1% testosterone (prepared with 50% ethanol) daily to establish a testosterone-induced pathological hair loss model; mice in the Control group were treated with 50% ethanol. Thirty minutes after testosterone application, the mice were administered the drug according to their groups (the experimental groups were given compound 3 by gavage at doses of 30 mg / kg and 60 mg / kg, respectively), while the Control group and Model group were given an equal volume of physiological saline, once daily for 21 consecutive days (see [link to relevant documentation]). Figure 2 ).

[0278] Depend on Figure 2 As can be seen, observations were made on days 15 and 21 after drug administration. Specifically, hair growth in mice on day 21 was as follows: Figure 2 As shown in the figure. 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) doses was significantly higher than that of the Model group.

[0279] Based on the above description, those skilled in the art can easily determine the basic features of the present invention, and various changes and modifications can be made to the present invention to suit various uses and conditions without departing from its spirit and scope.

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

Claims

1. A 1,3,5-trisubstituted indole compound, characterized in that: The compounds are those represented by general formula I and their enantiomers, and pharmaceutically acceptable salts: in, L is , , , ; M is or ; Where X is O or N, Y is C or N, and Z is CH, O or N, so that X, Y, Z together with the carbon and nitrogen atoms attached to them form a five-membered aromatic heterocyclic system; Q is CH2 or ; W stands for methylene; n is 0 or 1; R1 and R2 can be independently selected from hydrogen, (C3~C8) cycloalkyl, (C6~C8) cycloalkyl, or (C6~C8) cycloalkyl. 10 ) aryl, 4-10 heterocyclic, 5-10 heteroaryl; Alternatively, R1 and R2 together with the N atom to which they are attached form a 4- to 10-membered heterocycle containing at least one heteroatom, wherein the heterocycle is optionally substituted with 1 to 3 identical or different of the following groups: halogen, cyano, nitro, hydroxyl, mercapto, carboxyl, (carbon)-oxo, (sulfo)-oxo, (sulfo)-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 an unsubstituted (C1~C6) alkyl (C3~C8) cycloalkyl group or substituted with at least one of the following substituents: hydrogen, halogen, hydroxyl, cyano, methoxy, carboxyl, or amino. R4 is C6~C 10 Aryl, 5-10 heteroaryl; wherein the aryl, heteroaryl may be optionally and independently substituted by 1-4 identical or different of the following groups: halogen, cyano, nitro, hydroxyl, mercapto, carboxyl, (C1-C6)alkoxy, (C1-C6) acyl, (C2-C6) acyloxy, (C1-C6) alkoxycarbonyl, (C1-C4) alkylsulfonyl, (C1-C4) alkylsulfonyl.

2. The 1,3,5-trisubstituted indole compound according to claim 1, characterized in that: The compounds are those represented by general formula I and their enantiomers, or pharmaceutically acceptable salts; In the formula: L is , , , ; M is or ; X is O or N, Y is C or N, and Z is CH, O or N, so that X, Y, Z together with the carbon and nitrogen atoms attached to them form a five-membered aromatic heterocyclic system; Q is CH2 or ; W stands for methylene; n is 0 or 1; R1 and R2 can be the same or different and can be hydrogen, (C3~C6)cycloalkyl, (C6~C6)cycloalkyl, or other independent compounds. 10 aryl; Alternatively, R1 and R2 together with the N atom to which they are attached form a 4- to 10-membered heterocycle containing at least one heteroatom, wherein the heterocycle is optionally substituted with 1 to 2 identical or different of the following groups: halogen, cyano, hydroxyl, carboxyl, (carbon)-oxo, (sulfo)-dioxo, (C1-C4) alkyl, methoxy or hydroxy-substituted (C1-C4) alkyl, (C1-C4) alkoxy, (C2-C4) acyloxy, (C1-C4) alkoxycarbonyl; R3 is an unsubstituted or substituted structure consisting of one or two independently selected groups of hydrogen, halogen, hydroxyl, cyano, methoxy, or amino groups: (C1-C4) straight-chain or branched alkyl, (C3-C6) cycloalkyl; R4 is phenyl, naphthyl, or a 5-6 membered heteroaryl group, wherein the phenyl, naphthyl, or heteroaryl group is optionally and independently substituted by 1-3 identical or different groups of the following: halogen, cyano, nitro, hydroxyl, carboxyl, (C1-C4)alkoxy, (C2-C4)acyl, (C2-C4)acyloxy, or (C1-C4)alkoxycarbonyl.

3. The 1,3,5-trisubstituted indole compound according to claim 2, characterized in that: The compounds are those represented by general formula I and their enantiomers, or pharmaceutically acceptable salts; In the formula: L is , , , ; M is , or ; Q is CH2 or ; n is 0 or 1; R1 and R2 may be the same or different and are selected from hydrogen, (C3~C6) cycloalkyl, and phenyl. Alternatively, R1 and R2 together with the N atom to which they are attached form a 4- to 7-membered heterocycle containing at least one heteroatom, wherein the heterocycle is optionally substituted by 1 to 2 identical or different of the following groups: (C1 to C4) straight-chain or branched alkyl, (carbon)-oxo, (sulfo)-dioxo, hydroxy, hydroxymethyl, hydroxyethyl, carboxyl. R3 is a (C1~C4) alkyl or (C3~C6) cycloalkyl; R4 is a phenyl or pyridyl group, which may be optionally and independently substituted by 1 to 3 identical or different groups of the following: halogen, cyano, hydroxyl, carboxyl, methoxy.

4. The 1,3,5-trisubstituted indole compound according to claim 3, characterized in that: The compounds are those represented by general formula I and their enantiomers, or pharmaceutically acceptable salts; In the formula: L is , , , ; M is , or ; Q is CH2 or n = 0 or 1; R1 and R2 can be the same or different and are selected from hydrogen, methyl, ethyl, cyclopropyl, cyclohexyl, and phenyl. Alternatively, R1 and R2 together with the nitrogen atom to which they are attached form a 4- to 6-membered heterocycle containing at least one heteroatom; said heterocycle may optionally be substituted with at least one C1- to C3 alkyl, hydroxyl, carboxyl, or (thio)-dioxo group; R3 represents methyl or ethyl; R4 is a phenyl or pyridyl group substituted with the following: , , , , , , , , , , , or .

5. The 1,3,5-trisubstituted indole compound according to claim 4, characterized in that: The compounds are those represented by general formula I and their enantiomers, or pharmaceutically acceptable salts; In the formula: L is , , , ; M is , or ; Q is CH2 or n = 0 or 1; R1 and R2 can be the same or different and are selected from hydrogen, methyl, ethyl, cyclopropyl, cyclohexyl, and phenyl. Alternatively, R1 and R2 together with the nitrogen atoms they are attached to form... , , , , , , , , , , , or ; R3 represents methyl or ethyl; R4 is , , , , , , , , , , , or .

6. The 1,3,5-trisubstituted indole compound according to any one of claims 1 to 5, characterized in that: The compound is one of the following compounds and their enantiomers, or pharmaceutically acceptable salts; 3,4,5-Trichlorobenzyl(S)-(1-ethyl-5-(4-((2-(hydroxymethyl)pyrrolidone-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)pyrrolidone-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-indole-3-yl)carbamate; 1-(4-Fluorobenzyl)-3-(1-Methyl-5-(4-(morpholinylmethyl)-1H-1,2,3-triazol-1-yl)-1H-indol-3-yl)urea; 3,5-Dimethoxybenzyl (1-ethyl-5-(5-(morpholinylmethyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate; 1-(1-Ethyl-5-(5-(tetrahydropyrrolo-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)tetrahydropyrrolo-1-yl)methyl)-1,2,4-oxadiazol-3-yl)-1H-indol-3-yl)carbamate; 3,5-Dichlorobenzyl(R)-(1-ethyl-5-(5-((2-(hydroxymethyl)tetrahydropyrrolo-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-hydroxyazacyclobutane-1-yl)-2-oxoethyl)piperazin-1-yl)-1H-indole-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 of formula I as claimed in claim 1, its enantiomers, and pharmaceutically acceptable salts.

8. Use of a 1,3,5-trisubstituted indole compound of claim 1 or a pharmaceutical composition of claim 7 in the preparation of an ATX inhibitor.

9. Use of a 1,3,5-trisubstituted indole compound of claim 1 or a pharmaceutical composition of claim 7 in the preparation of a medicament having therapeutic or preventative effects on tumors, fibrotic diseases, and promoting hair growth in mammals.

10. The application according to claim 9, characterized in that, The tumor is breast cancer or glioma, and the fibrotic disease is pulmonary fibrosis, liver fibrosis, or myocardial fibrosis.

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