CN103784450B - Small molecular compound for inhibiting Src non-receptor tyrosine kinase and its pharmaceutical use - Google Patents

Abstract

The invention belongs to field of biological pharmacy, relate to the new framework types micromolecular compound and medicinal usage thereof that suppress Src nonreceptor tyrosine kinase.Is the present invention by using Accelrys? Discovery? Studio? the Pharmacophore Model of 3.5 structure Src inhibitors of kinases, and with this Pharmacophore Model for queries searches known chemicals storehouse, obtain the compound of formula I-V; Further by biological activity test, result confirms, the micromolecular compound obtained has comparatively high inhibition effect to Src nonreceptor tyrosine kinase; Compound of the present invention and medicine salt thereof or hydrate can be used for preparing antitumor drug, the application particularly in the tumor disease that treatment is relevant to Src.Described compound can be used as lead compound, carries out structure of modification, prepares and synthesize new Src inhibitors of kinases.

Description

Small molecular compound for inhibiting Src non-receptor tyrosine kinase and its pharmaceutical use

technical field

The invention belongs to the field of biopharmaceuticals, and relates to a new skeleton-type small molecular compound for inhibiting Src non-receptor tyrosine kinase and its medicinal application. The small molecular compound has strong inhibitory effect on Src non-receptor tyrosine kinase and can be used to prepare anti-tumor drugs, especially in the preparation of drugs for treating tumor diseases related to Src.

Background technique

The prior art discloses that the Src gene is the first oncogene discovered, and its expression product, the Src protein, is a tyrosine kinase whose main function is to catalyze the phosphorylation of tyrosine residues in downstream signaling proteins, thereby activating signal transduction. conduction path. Although Src was originally found to exist in Rous sarcoma retrovirus (retrovirusRoussarcomavirus), subsequent studies have found that there is a highly homologous c-Src protein kinase in cells. Studies in recent years have shown that Src kinase overexpression is related to the occurrence and metastasis of various human tumors, such as chronic myeloid leukemia, lung cancer, colon cancer, breast cancer and pancreatic cancer (RosalynB.IrbyandTimothyJ.Yeatman, Oncogene (2000) 19, 5636-5642). Therefore, inhibiting the excessive tyrosine kinase activity of Src to block the tumor signal transduction pathway mediated by Src is a potential anti-tumor method.

At present, the Src small molecule inhibitor developed by Bristol-Myers Squibb, whose general name is dasatinib, was approved by the US FDA in 2006 for the treatment of chronic myeloid leukemia (CML). It is also reported that the small molecule inhibitors of Src, namely bosutinib and saracatinib, have entered the stage of clinical research. Although a large number of Src kinase inhibitors have been reported, the structural diversity of these compounds is still not high, so it is difficult to overcome the problem of drug resistance of tumors. The present invention aims at discovering the Src kinase inhibitor with new skeleton type by using the method of computer-aided drug design.

Contents of the invention

The purpose of the present invention is to provide a new skeleton type small molecule compound with strong inhibitory effect on Src non-receptor tyrosine kinase, specifically related to 2-(quinazoline-4-amino)-5-thiazolecarboxamide derivatives and its medicinal uses. In particular, it relates to the application of the new skeleton type small molecular compound in the preparation of anti-tumor drugs, especially in the preparation of drugs for the treatment of tumor diseases related to Src.

The pharmacophore-based virtual screening of the present invention is an efficient structure-based drug design method. This method is based on the basic principles of computational chemistry, using high-speed computer software and hardware as tools, combined with biological activity tests to quickly discover lead compounds, and has the advantages of low cost and high efficiency. The present invention constructs the pharmacophore model of Src kinase inhibitor by using AccelrysDiscoveryStudio3.5, and uses the pharmacophore model as the query structure to search the known chemical library, and obtains the new skeleton type small molecule compound, and it is confirmed by experiments that the The small molecular compound of the new skeleton type has a strong inhibitory effect on Src non-receptor tyrosine kinase, and can be further used to prepare antitumor drugs, especially for the preparation of tumor drugs caused by the regulation disorder of the Src kinase signal transduction system.

Specifically, the 2-(quinazoline-4-amino)-5-thiazolecarboxamide derivative compound of the present invention has the structure of formula I, formula II, formula III, formula IV or formula V.

More specifically, the compounds of the formula I structure in the present invention include their pharmaceutically acceptable salts and hydrates:

In the formula

R ₁ : hydrogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ substituted alkyl, C ₁ -C ₄ alkoxy, aryl, substituted aryl, acyl containing 1-4 carbons;

The compound of the formula II structure of the present invention, including its pharmaceutically acceptable salt and hydrate:

In the formula

R ₁ : hydrogen, halogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ substituted alkyl,

R ₂ : hydrogen, halogen, aryl, substituted aryl,

R ₃ : hydrogen, halogen, hydroxyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, cyano, acyl with 1-4 carbons,

R ₃ : hydrogen, halogen, hydroxyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, cyano, acyl containing 1-4 carbons;

The compound of the formula III structure of the present invention, including its pharmaceutically acceptable salts and hydrates:

III

In the formula

R ₁ : hydrogen, halogen, hydroxyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, cyano, acyl with 1-4 carbons, aryl, hetero Aryl,

R ₂ : hydrogen, halogen, hydroxyl, cyano,

R ₃ : hydrogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy;

Compounds of the formula IV structure of the present invention, including their pharmaceutically acceptable salts and hydrates:

In the formula

R ₁ : hydrogen, halogen, hydroxyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, cyano, acyl with 1-4 carbons, aryl, hetero Aryl,

R ₂ : hydrogen, halogen, hydroxyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, cyano, acyl with 1-4 carbons, aryl, hetero Aryl;

Compounds of the formula V structure of the present invention, including their pharmaceutically acceptable salts and hydrates:

In the formula

R ₁ : hydrogen, hydroxyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, acyl with 1-4 carbons, aryl, substituted aryl, heteroaryl base, substituted heteroaryl,

R ₂ : hydrogen, hydroxyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, acyl with 1-4 carbons, aryl, substituted aryl, heteroaryl base, substituted heteroaryl.

The present invention uses AccelrysDiscoveryStudio3.5 to construct the pharmacophore model of the Src kinase inhibitor, and uses the pharmacophore model as a query structure to search the known chemical library to obtain the compound of formula I-V; further through the biological activity test, the results confirm that, The obtained small molecular compound of the new skeleton type of formula I-V has a strong inhibitory effect on Src non-receptor tyrosine kinase; further, the compound of formula I-V described in the present invention and its medicinal salt or hydrate can be used to prepare anti-tumor Medicine, especially the application in the treatment of tumor diseases related to Src. Furthermore, the compounds of formulas I-V described in the present invention can be used as lead compounds for structural modification to prepare and synthesize new Src kinase inhibitors.

For ease of understanding, the present invention will be described in detail below through specific drawings and embodiments. It should be pointed out that the specific examples and accompanying drawings are only for illustration. Obviously, those skilled in the art can make various amendments and changes within the scope of the present invention according to the description herein. These amendments and Modifications are also included within the scope of the present invention.

Description of drawings

Figure 1 shows the crystal structure of Src kinase and its binding pocket (PDBCode: 2H8H).

Figure 2 is the pharmacophore model of Src kinase inhibitors.

detailed description

Embodiment 1 is based on the virtual screening of DOCK program and GOLD program

The crystal structure of Src kinase was obtained from the ProteinDataBank protein database (PDBCode: 2H8H), and the crystal structure of the complex was preprocessed with the Chimera program, that is, removing water molecules and adding hydrogen atoms. Through the AccelrysDiscoveryStudio program, the pharmacophore model was constructed based on the crystal complex and the exclusion volume was added;

The 2-(quinazoline-4-amino)-5-thiazole carboxamide derivative compound having the structure of formula I, formula II, formula III, formula IV or formula V of the present invention is obtained.

Wherein, the compound of the formula I structure includes its pharmaceutical salt and hydrate:

In the formula

R ₁ : hydrogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ substituted alkyl, C ₁ -C ₄ alkoxy, aryl, substituted aryl, acyl containing 1-4 carbons;

The compound of the formula II structure, including its available pharmaceutical salts and hydrates:

In the formula

R ₁ : hydrogen, halogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ substituted alkyl,

R ₂ : hydrogen, halogen, aryl, substituted aryl,

R ₃ : hydrogen, halogen, hydroxyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, cyano, acyl with 1-4 carbons,

R ₃ : hydrogen, halogen, hydroxyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, cyano, acyl containing 1-4 carbons;

The compound of formula III structure, including its pharmaceutical salt and hydrate:

In the formula

R ₁ : hydrogen, halogen, hydroxyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, cyano, acyl with 1-4 carbons, aryl, hetero Aryl,

R ₂ : hydrogen, halogen, hydroxyl, cyano,

R ₃ : hydrogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy;

The compound of formula IV structure, including its available salt and hydrate:

In the formula

R ₁ : hydrogen, halogen, hydroxyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, cyano, acyl with 1-4 carbons, aryl, hetero Aryl,

R ₂ : hydrogen, halogen, hydroxyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, cyano, acyl with 1-4 carbons, aryl, hetero Aryl;

Compounds of the formula V structure, including their pharmaceutically acceptable salts and hydrates:

In the formula

R ₁ : hydrogen, hydroxyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, acyl with 1-4 carbons, aryl, substituted aryl, heteroaryl base, substituted heteroaryl,

R ₂ : hydrogen, hydroxyl, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ sulfonyl, acyl with 1-4 carbons, aryl, substituted aryl, heteroaryl base, substituted heteroaryl.

Embodiment 2 biological activity test

Seven candidate compounds were selected from the Enamine small molecule database (http://www.asinex.com), purchased and tested for activity.

All compounds were tested for their inhibition of Src kinase activity by Electrophoretic Mobility Shift Assay (EMSA), and the experiments were carried out on 384-well plates.

In the test, the positive control substance of the Src kinase inhibitor was Staurosporine, and both the test drug and the positive drug were dissolved in DMSO and mixed with the kinase buffer (20mM HEPES, pH7.5, 0.01%TritonX-100, 5mMMnCl2, 2mMDTT), and shaken Mix on the mixer for 10 minutes, then add Src kinase to the wells and incubate at room temperature for 10 minutes; add phosphoacceptorpeptide FAM-P4 and ATP containing tyrosine to the wells to finally obtain an enzyme reaction system of 25uL/well, and incubate at 28°C 60min. At the end of the incubation, add 25uL of stop buffer (100mMHEPES, pH7.5, 0.015%Brij-35, 0.2%CoatingReagent#3) to the empty wells; use the Caliper detection system to read the data, measure two tubes for each concentration, and test each compound Two independent experiments. Compounds 1-5 among the candidate compounds showed high inhibitory activity on Src kinase, and their chemical structures and IC ₅₀ values are shown in Table 1.

Table 1 Chemical structures and IC ₅₀ values of compounds 1-5

The results show that the compounds 1-5 have obvious inhibitory effect on Src kinase. Based on the fact that Src kinase is closely related to various tumor diseases, the compounds involved in the present invention can further prepare anti-tumor drugs, and block the Src-mediated tumor signal transduction pathway by inhibiting the excessive tyrosine kinase activity of Src, To achieve the purpose of treating tumors; the tumors described in the present invention include chronic myeloid leukemia, lung cancer, colon cancer, breast cancer and pancreatic cancer.

Claims (6)

1. the purposes of the 2-(quinazoline-4-amino)-5-thiazolecarboxamide derivative compound of formula I in the preparation of antineoplastic drugs; Wherein, the compound of the formula I structure includes its available pharmaceutical salts and hydrates: In the formula R ₁ : hydrogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ substituted alkyl, C ₁ -C ₄ alkoxy, aryl, substituted aryl, acyl having 1-4 carbons. 2. Use of the compound of formula I described in claim 1 in the preparation of Src kinase inhibitors. 3. Use of the compound of formula I described in claim 1 in the preparation or synthesis of a lead compound of an Src kinase inhibitor. 4. The use according to claim 1, characterized in that said compound inhibits the activity of Src kinase. 5. according to the described purposes of claim 1, it is characterized in that, described compound is: 6. The use according to claim 1, wherein the tumor is chronic myeloid leukemia, lung cancer, colon cancer, breast cancer or pancreatic cancer.