CN109303777B

CN109303777B - Application of imidazole [4,5-b ] pyridine mercaptoacetamide derivative in preparation of anti-human solid tumor drugs

Info

Publication number: CN109303777B
Application number: CN201810800211.1A
Authority: CN
Inventors: 展鹏; 刘新泳; 宋宇宁; 武高禅; 李潇
Original assignee: Shandong University
Current assignee: Shandong University
Priority date: 2018-07-20
Filing date: 2018-07-20
Publication date: 2020-04-17
Anticipated expiration: 2038-07-20
Also published as: CN109303777A

Abstract

The invention relates to an imidazopyridine mercaptoacetamide derivative shown as a general formula I or a medicinal salt thereof, a medicinal composition containing the compound and application of the medicinal composition in preparation of a medicament for resisting human solid tumors, wherein the human solid tumors comprise human large cell lung cancer, human liver cancer, human colon cancer and human breast cancer.

Description

Application of imidazole [4,5-b ] pyridine mercaptoacetamide derivative in preparation of anti-human solid tumor drugs

Technical Field

The invention relates to application of imidazole [4,5-b ] pyridine mercaptoacetamide derivatives or pharmaceutically acceptable salts, esters or prodrugs thereof in preparation of anti-human solid tumor drugs, and belongs to the technical field of medicines.

Background

Malignant tumors are common and frequently occurring diseases that currently pose a serious threat to human health and survival. According to the reports of the world health organization, the number of patients dying from cancer every year worldwide is tens of millions, and the incidence rate of tumors, especially malignant tumors, and the number of people dying from cancer every year are increasing year by year due to the aggravation of environmental pollution and poor living habits in recent years. In the treatment of malignant tumors, drug therapy plays an important role. Although there are dozens of common antitumor drugs in clinical use, there are still many problems, such as poor water solubility, severe toxic and side effects, and easy generation of acquired drug resistance, which cannot meet the needs of clinical treatment. Especially, the treatment of solid tumors which are the most serious and account for more than 90 percent of malignant tumors and endanger the life health of human beings, such as lung cancer, liver cancer, colon cancer, breast cancer and the like, does not achieve satisfactory effect. Therefore, the research on the antitumor drugs with good curative effect and small side effect is still one of the hot spots of the new drug research.

The discovery of the active lead of the medicine is the source and guarantee of the development of new medicines and is a key link of the whole new medicine research and development chain. The construction of compound libraries with various structures and high quality has important significance for the discovery and structure optimization of lead compounds. Combinatorial chemistry techniques can rapidly synthesize a large number of compounds, but the resulting compound libraries lack structural diversity features. Although the compound libraries obtained by the diversity synthesis strategy have molecular diversity, the drug-like properties are often poor. How to obtain a high-quality compound library with drug-like properties and diversity has become a bottleneck in drug research and development, and is also an important research task in the field of medicinal chemistry. Dominant structures refer to molecular backbones from which ligands with high affinity for multiple targets can be derived. The ideal advantageous structure is suitable for introducing different functional groups, and enriches the diversity of compound structures and activities so as to improve the activity, selectivity and druggability.

The literature research finds that the heterocyclic mercaptoacetamides compounds have wide biological activity, and can be used as a basic structure mother nucleus for forming a pharmacophore so as to meet the space requirement of a drug special action target; may also be part of an active substituent or ring system to produce a corresponding biological activity; in addition, the compound has better drug-like characteristics and biocompatibility. Therefore, they are widely used as the dominant heterocyclic structure in the construction and structure optimization of compound libraries.

Phenotypic drug screening is a drug screening method based on cell disease models and other organism phenotypes, and is an important way for discovering drug activity lead compounds with brand-new structural frameworks. Therefore, by taking the heterocyclic mercaptoacetamide dominant structure as a template, constructing an active focusing compound library for phenotype screening, the method has important significance for discovering a new generation of antitumor drugs with independent intellectual property rights.

Chinese patent document CN105175414A discloses imidazole [4,5-b ] pyridine mercaptoacetamide derivatives (general formula I) and a preparation method and application thereof, and only discloses the application of the imidazole [4,5-b ] pyridine mercaptoacetamide derivatives in the preparation of drugs for preventing Human Immunodeficiency Virus (HIV) infection and resisting leukemia, which is a tumor disease completely different from human solid tumors, and the application of the compounds in other aspects is not found in the prior art.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides an in-vitro anti-tumor phenotypic activity screening result and application of an imidazole [4,5-b ] pyridine mercaptoacetamide derivative, and particularly relates to the application of the imidazole [4,5-b ] pyridine mercaptoacetamide derivative in preparation of anti-human solid tumor drugs.

The technical scheme of the invention is as follows:

application of imidazole [4,5-b ] pyridine mercaptoacetamide derivatives in preparation of anti-human solid tumor drugs; the imidazole [4,5-b ] pyridine mercaptoacetamide derivative has a structure shown in the following general formula I:

wherein, in the general formula I,

ar directly attached to the pyrimidine ring₁Is a substituted benzene ring, a naphthalene ring or a nitrogen-containing aromatic heterocycle with diverse structures; preferably, Ar is₁Is a 2, 4-dichlorobenzene ring, a 2, 4-dibromobenzene ring, a 2-chloro-4-cyclopropylbenzene ring, a 2-bromo-4-cyclopropylbenzene ring, a 2-chloro-4-tert-butylbenzene ring, a 2-bromo-4-tert-butylbenzene ring, a 1-naphthalene ring, a 2-naphthalene ring, a 4-cyclopropyl-1-naphthalene ring or a 4-tert-butyl-1-naphthalene ring;

by chain-linked Ar₂Is a substituted benzene ring or a nitrogen-containing six-membered aromatic heterocycle; preferably, X is N or C; r₁Is H, F, Cl, Br or NO₂；R₂Is H, CN, OMe, COMe, SO₂NH₂、COOH、CONH₂、COOMe、COOEt、CONHOH、CONHOMe、CONHCH₂COOEt or CONHCH₂(CH₃)-COOEt；

R on the mercaptoacetamide chain₃、R₄Each independently is H, (C)_1-4) Alkyl or halogen; preferably, R₃、R₄Is H or F, R₃And R₄One is H and the other is selected from CH₃Or F.

According to the present invention, it is further preferred that the compound of formula I is one of the compounds of the following structure:

compound No. 13: 3-bromo-4- (2- ((1- (4-cyclopropylnaphthalene 1-yl) -1H-imidazo [4,5-b ] pyridin-2-yl) mercapto) acetamide) benzamide;

compound No. 18: ethyl (3-chloro-4- (2- ((1- (4-cyclopropylnaphthalene 1-yl) -1H-imidazo [4,5-b ] pyridin-2-yl) mercapto) acetamide) benzoyl) glycine;

compound No. 20: 3-chloro-4- (2- ((1- (4-cyclopropylnaphthalen-1-yl) -1H-imidazo [4,5-b ] pyridin-2-yl) mercapto) acetamide) benzamide.

According to the application of the imidazo [4,5-b ] pyridine mercaptoacetamide derivative in preparing the anti-human solid tumor medicine, the imidazo [4,5-b ] pyridine mercaptoacetamide derivative also comprises pharmaceutically acceptable salts, esters or prodrugs of the imidazo [4,5-b ] pyridine mercaptoacetamide derivative, and a pharmaceutical composition containing the compound.

The application of the imidazo [4,5-b ] pyridine mercaptoacetamide derivative in preparing the anti-human solid tumor medicament is disclosed, wherein the human solid tumor comprises human large cell lung cancer, human liver cancer, human colon cancer and human breast cancer.

According to the application of the imidazo [4,5-b ] pyridine mercaptoacetamide derivative in preparing the anti-human solid tumor medicament, the medicament form is any clinically or pharmaceutically acceptable form. Preferably, the dosage form of the medicament is tablets, capsules, solutions, suspensions, granules or injections.

Description of terms:

the term "pharmaceutically acceptable salt" as used herein means that a salt of a compound which is, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and lower animals without undue toxicity, irritation, allergic response and the like, commensurate with a reasonable benefit/risk ratio, generally water or oil soluble or dispersible, and effective for its intended use. Including pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts, which are useful herein and which are compatible with the chemical nature of the compounds of formula I. A list of suitable salts is found on pages 1-19 of s.m. berge et al, j.pharm.sci.,1977, 66.

The term "prodrug" as used in the present invention refers to pharmaceutically acceptable derivatives such that the resulting biotransformation product of these derivatives is the active drug as defined for the compound of formula I. Examples of such derivatives include, but are not limited to, esters and amides.

Has the advantages that:

the in vitro anti-tumor activity screening experiment shows that the imidazole [4,5-b ] pyridine mercaptoacetamide derivative of the compound shown in the formula I has the activity of inhibiting cell proliferation to different degrees on 4 cells such as a human large cell lung cancer cell strain NCI-H460, a human liver cancer cell Bel-7402, a human colon cancer cell SW620, a human breast cancer cell strain MDA-MB-231 and the like. Especially compound 13, compound 18, compound 20, achieved unexpected results. Therefore, the compound shown in the formula I can be used for preparing anti-human solid tumor medicaments, and has important research and development prospects.

Detailed Description

The following examples are given to aid in the understanding of the invention, but are not intended to limit the scope of the invention.

Example 1: antitumor Activity test (CCK-8 method)

Purpose of the experiment:

the in vitro cytotoxic activity and the in vitro cytotoxic strength of the compound on 4 cells such as a human large cell lung cancer cell strain NCI-H460, a human liver cancer cell Bel-7402, a human colon cancer cell SW620 and a human breast cancer cell strain MDA-MB-231 are detected.

Experimental materials:

96-well cell culture plates were purchased from Costar corporation;

RPMI 1640 medium was purchased from Gibco;

DMEM medium was purchased from Gibco;

fetal bovine serum was purchased from Gibco;

DMSO was purchased from Sigma;

EnoGeneCell^TMa Counting Kit-8(CCK-8) cell viability assay Kit (E1CK-000208-10) was purchased from Nanjing Enjing Biotech Ltd.

An experimental instrument:

a ChemBase CBS-CJ-1FD super clean bench,

MCO-15AC carbon dioxide incubator, Sanyo SANYO, Japan,

XD-202 fluorescence inverted biomicroscope, Yongxin optics Limited in south Jing Jiangnan,

thermo MK3 microplate reader, thermoelectricity, usa.

The tested drugs are:

the invention relates to compounds of formula I. Samples are prepared into mother liquor by DMSO, and are diluted by incomplete culture medium when in use, and the final concentration of DMSO does not exceed 0.01%.

Positive control: paclitaxel injection, tai chi group sichuan tai chi pharmaceutical co ltd, 5mL:30mg, batch No.: 14121201.

cell lines:

human large cell lung cancer cell strain NCI-H460 and human liver cancer cell Bel-7402, wherein the cells are purchased from Nanjing Enjinjing Biotech limited company and cultured in RPMI 1640 medium containing 10% fetal bovine serum;

SW620 of human colon cancer cells and MDA-MB-231 of human breast cancer cell strains, which are purchased from Nanjing Enjing Biotech limited and cultured in DMEM medium containing 10% fetal bovine serum.

The experimental scheme is as follows:

CCK-8 staining method for detecting cell viability

The experiment was carried out using cells with a viable cell fraction of more than 90%. Cell proliferation inhibition assay Using EnoGeneCell^TMCounting Kit-8(CCK-8) cell viability detection Kit. Digesting and counting cells to obtain a concentration of 1 × 10⁵Cell suspension/mL, 100. mu.L of cell suspension per well in 96-well plates (1X 10 per well)⁴Individual cells); the 96-well plate was placed at 37 ℃ in 5% CO₂Culturing in an incubator for 24 hours; adding 100 μ L of corresponding culture medium containing medicine into each well, and simultaneously setting up a negative control group, a solvent control group, a positive control group, and 5 multiple wells per group; the 96-well plate was placed at 37 ℃ in 5% CO₂Culturing for 72 hours in an incubator; adding 10 μ L CCK-8 solution into each well, incubating the culture plate in incubator for 4 hr, measuring OD value at 450nm with microplate reader, and calculating compounds for human large cell lung cancer cell strain NCI-H460 and human liver cancerInhibition rate and IC of 6 cells such as cell Bel-7402, human colon cancer cell SW620 and human breast cancer cell strain MDA-MB-231₅₀The value is obtained.

Dose setting:

dose 1 group: 100 mu M;

dose 2 groups: 50 mu M;

dose 3 groups: 25 mu M;

dose 4 groups: 12.5 mu M;

dose 5 groups: 6.25 mu M;

dose 6 groups: 3.125 μ M;

dose 7 groups: 1.5625 μ M;

dose 8 groups: 0.78125 μ M;

dose 9 groups: 0.390625 μ M;

dose 10 groups: 0.1953125 μ M;

positive control: 10. mu.g/mL.

The experimental results are as follows: the compound of the formula I has activity of inhibiting cell proliferation to different degrees on 4 cells such as a human large cell lung cancer cell strain NCI-H460, a human liver cancer cell Bel-7402, a human colon cancer cell SW620, a human breast cancer cell strain MDA-MB-231 and the like. The results for the 3 compounds with the best activity are detailed in tables 1-5 below.

TABLE 1 inhibition ratio of in vitro proliferation Activity of 3 Compounds on human Large cell Lung cancer cell line NCI-H460 and IC₅₀The value is obtained. (unit:%)

TABLE 2 inhibition ratio and IC of in vitro proliferation activity of 3 compounds on human hepatoma cell Bel-7402₅₀The value is obtained. (unit:%)

TABLE 3 inhibition of in vitro proliferative Activity of 3 Compounds on human Colon cancer cells SW620 and IC₅₀The value is obtained. (unit:%)

TABLE 4 inhibition ratio and IC of 3 compounds on in vitro proliferation activity of human breast cancer cell line MDA-MB-231₅₀The value is obtained. (unit:%)

TABLE 5 IC of 3 Compounds on individual cells₅₀Value summary (Unit: mu M)

The experimental results show that: the 3 compounds have significant cell proliferation inhibiting activity on 4 solid tumor cells such as human large cell lung cancer cell strain NCI-H460, human liver cancer cell Bel-7402, human colon cancer cell SW620, human breast cancer cell strain MDA-MB-231 and the like.

The 18 # compound and the 20 # compound have obvious cytotoxic activity and IC activity on human large cell lung cancer cell strain NCI-H460₅₀The values are respectively: 10.53 μ Μ and 10.6 μ Μ; the compound No. 13 shows certain cytotoxic activity on human large cell lung cancer cell strain NCI-H460, and IC thereof₅₀The values are: 23.73 μm.

The compound No. 18 has obvious cytotoxic activity to human hepatoma cell Bel-7402,IC thereof₅₀The values are: 13.41 μm; the cytotoxic activity of the compound No. 13 and the compound No. 20 on human hepatoma cell Bel-7402 is relatively low, and the IC50 values are respectively as follows: 54.6 μm and 79.58 μm.

Compound No. 13, compound No. 18 and compound No. 20 have obvious cytotoxic activity on human colon cancer cells SW620, and IC of the compound₅₀The values are respectively: 27.83 μm, 23.84 μm and 18.35 μm.

The 18 # compound and the 20 # compound have obvious cytotoxic activity and IC activity on human breast cancer cell strains MDA-MB-231₅₀The values are respectively: 11.95 μ Μ and 11.99 μ Μ; the compound No. 13 shows certain cytotoxic activity against human breast cancer cell strain MDA-MB-231, and IC thereof₅₀The values are: 35.72 μm.

The in vitro cytotoxic activity intensity sequence of the 3 compounds on the human large cell lung cancer cell strain NCI-H460 is as follows: compound No. 18 > compound No. 20 > compound No. 13. The in vitro cytotoxic activity intensity sequence of 3 compounds on human hepatoma cell Bel-7402 is as follows: compound No. 18 > compound No. 13 > compound No. 20. The in vitro cytotoxic activity of 3 compounds on human colon cancer cells SW620 has the following sequence: compound No. 20 > compound No. 18 > compound No. 13. The in vitro cytotoxic activity intensity sequence of the 3 compounds on the human breast cancer cell strain MDA-MB-231 is as follows: compound No. 18 > compound No. 20 > compound No. 13.

Therefore, the imidazole [4,5-b ] pyridine mercaptoacetamide derivative with the general formula I can be used for preparing anti-human solid tumor drugs.

While the preferred embodiments of the present invention have been illustrated and described, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. Application of imidazole [4,5-b ] pyridine mercaptoacetamide derivatives in preparation of anti-human solid tumor drugs;

the imidazole [4,5-b ] pyridine mercaptoacetamide derivative is one of the following compounds:

compound No. 20: 3-chloro-4- (2- ((1- (4-cyclopropylnaphthalen-1-yl) -1H-imidazo [4,5-b ] pyridin-2-yl) mercapto) acetamide) benzamide;

the human solid tumor is human large cell lung cancer, human liver cancer, human colon cancer and human breast cancer.

2. The use according to claim 1, wherein said imidazo [4,5-b ] pyridine mercaptoacetamide derivatives further comprise pharmaceutically acceptable salts, esters or prodrugs of imidazo [4,5-b ] pyridine mercaptoacetamide derivatives, as well as pharmaceutical compositions containing said compounds.

3. The use of claim 1, wherein the medicament is in any form that is clinically or pharmaceutically acceptable.

4. The use according to claim 3, wherein the medicament is in the form of a tablet, capsule, solution, suspension, granule or injection.