Application of compound in inhibition of Aurora and JAK kinase family activity and treatment of several tumors

Abstract

The invention relates to the technical field of chemical medicines, in particular to application of a compound 2-(4-methanoyl) anilino-4-(1H-indole-4-oxy)-5-trifluoromethylpyrimidine in inhibition of the activity of Aurora and JAK kinase families and in treatment of several tumors. On the basis of the research of 2-(4-methanoyl) anilino-4-amino-5-trifluoromethylpyrimidine derivative and application (patent application number: 202110571344.8) in the early stage, the compound is found to be capable of inhibiting the activity of FAK kinase and effectively inhibiting the family activity of Aurora and JAK kinase, and the inhibition effects are respectively 100 times and 5 times or more superior to those of the FAK kinase, so that the compound can be used for preparing a novel compound for inhibiting the activity of the FAK kinase. The compound can be used as a dual-effect anti-tumor inhibitor and can reduce proliferation of breast cancer, lung cancer, leukemia, thyroid cancer, prostate cancer, glioma, ovarian cancer and lymphoma cells, the activity is not observed in other FAK kinase inhibitors (such as Defactinib), and the compound can be used as a dual-effect anti-tumor inhibitor. The compound is applied to preparation of medicines for treating breast cancer, lung cancer, leukemia, thyroid cancer, prostate cancer, glioma, ovarian cancer, lymphoma and idiopathic pulmonary fibrosis.

Description

Technical Field

[0001] The present invention relates to the technical field of chemical medicine, and in particular to the application of a compound in inhibiting the activity of Aurora and JAK kinase family and in treating several tumors. Background Art

[0002] The Aurora kinase family (A, B, and C) is a family of serine / threonine kinase isoforms that regulate mitosis and meiosis. They participate in chromosome segregation and cytokinesis, ensuring the correct attachment of chromosomes. Their dysfunction can lead to genetic instability and cell carcinogenesis. Studies have shown that Aurora kinases are often overexpressed in tumor cells and are potential targets for the development of anti-tumor drugs. In addition, Aurora kinases are also involved in cell cycle regulation and immune cell activation, regulating immune cell proliferation and function to treat autoimmune diseases, and are also associated with neurodegenerative diseases.

[0003] The Janus kinase (JAK) family is a group of small protein tyrosine kinases that are the core of intracellular signal transduction downstream of cytokine receptors. The dysregulated JAK3 / STAT signaling pathway is a hallmark of the development of human malignant tumors. High expression of JAK3 can activate the STAT1, 3, 5, and 6 pathways, while inhibition of the JAK3 / STAT pathway can inhibit tumor cell proliferation or induce apoptosis, exerting anti-tumor efficacy. JAK is not only related to tumor diseases, but also to autoimmune diseases, inflammatory diseases, and hematological diseases. JAK is particularly critical for immune cell-related cytokine receptor signal transduction, and has been regarded as a potential target for the treatment of inflammatory and autoimmune diseases with limited side effects. JAK inhibitors can regulate immune responses, reduce the invasion of inflammatory cells, reduce the release of inflammatory factors, and improve the bone marrow hematopoietic microenvironment and abnormal cell signals. The JAK-STAT pathway is associated with the occurrence of fibrosis. In systemic sclerosis, the STAT3 signaling pathway is overactivated, which promotes the further development of fibrosis and is closely related to idiopathic pulmonary fibrosis (IPF). In IPF patients, phosphorylated STAT3 has been detected in the nuclei of myofibroblasts, alveolar macrophages and alveolar epithelial cells, and inhibiting JAK3 activity can regulate the JAK / STAT pathway to improve pulmonary fibrosis. Summary of the invention

[0004] In order to solve the above problems, the present invention provides a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine for use in inhibiting the activity of Aurora and JAK kinase family and in treating breast cancer, lung cancer, leukemia, thyroid cancer, prostate cancer, glioma, ovarian cancer and lymphoma. The structural formula of the compound is shown in Formula I below:

[0005]

[0006] In addition to inhibiting the activity of FAK kinase, the compound can very effectively inhibit the activity of Aurora and JAK kinase families, and can be fully used to inhibit the activity of Aurora and JAK kinase families.

[0007] Furthermore, the compound can be used to prepare activity inhibitors of Aurora kinases AuroraA, Aurora B and Aurora C.

[0008] Furthermore, the compound can be used to prepare activity inhibitors of protein tyrosine kinases JAK 1, JAK 2 and JAK 3.

[0009] Furthermore, the compound can be used to prepare activity inhibitors of adenylate-activated protein kinase AMPKα1, tropomyosin receptor kinase TRKA, and mitogen-activated protein kinase (MAPK1, MAPK2, MAPK).

[0010] The compound can effectively weaken the survival rate of breast cancer cells, lung cancer cells, leukemia cells, thyroid cancer cells, prostate cancer cells, glioma cells, ovarian cancer cells and lymphoma cells, and its activity is significantly stronger than that of the FAK inhibitor Defactinib. Among them, the activity of compound 1 in inhibiting MDA-MB-231 breast cancer cells is more than 10 times that of Defactinib. In addition, no toxicity to normal embryonic kidney cells was observed at 20 μM, while Defactinib's IC 50 It is 3.4 μM, so the compound can be used to treat breast cancer, lung cancer, leukemia, thyroid cancer, prostate cancer, glioma, ovarian cancer, lymphoma, etc.

[0011] Furthermore, the compound can be used to prepare breast cancer drugs.

[0012] Furthermore, the compound can be used to prepare lung cancer drugs.

[0013] Furthermore, the compound can be used to prepare leukemia drugs, especially chronic myeloid leukemia.

[0014] Furthermore, the compound can be used to prepare thyroid cancer drugs.

[0015] Furthermore, the compound can be used for preparing prostate cancer drugs.

[0016] Furthermore, the compound can be used to prepare glioma drugs.

[0017] Furthermore, the compound can be used for preparing ovarian tumor drugs.

[0018] Furthermore, the compound can be used for preparing lymphoma drugs.

[0019] The JAK-STAT pathway is associated with the occurrence of fibrosis. In systemic sclerosis, the STAT3 signaling pathway is overactivated, which promotes the further development of fibrosis and is closely related to idiopathic pulmonary fibrosis (IPF). In IPF patients, phosphorylated STAT3 has been detected in the nuclei of myofibroblasts, alveolar macrophages, and alveolar epithelial cells, and inhibiting JAK3 activity can regulate the JAK / STAT pathway to improve pulmonary fibrosis. This compound can effectively inhibit JAK activity to further improve pulmonary fibrosis and can be fully used in the treatment of idiopathic pulmonary fibrosis.

[0020] Furthermore, the compound can be used for preparing drugs for idiopathic pulmonary fibrosis.

[0021] Compared with the prior art, the technical effects created by the present invention are embodied in:

[0022] (1) Based on the previous research on "2-(4-carboxamido)anilino-4-amino-5-trifluoromethylpyrimidine derivatives and applications" (patent application number 202110571344.8), the inventors of the present application found that in addition to inhibiting the activity of FAK kinase, the compound can also effectively inhibit the activity of Aurora and JAK kinase families, and the inhibitory effect is 100 times and 5 times better than FAK kinase, respectively, and can reduce the proliferation of breast cancer, lung cancer, leukemia, thyroid cancer, prostate cancer, glioma, ovarian cancer and lymphoma cells. This activity has not been observed in other FAK kinase inhibitors (such as Defactinib). The compound can be used as a dual-acting anti-tumor inhibitor and used in the preparation of drugs for the treatment of breast cancer, lung cancer, leukemia, thyroid cancer, prostate cancer, glioma, ovarian cancer, lymphoma and idiopathic pulmonary fibrosis.

[0023] (2) Compound 1 of the present invention is simple to prepare and has a high yield. The raw materials are cheap and easily available. It can target JAK3 and Aurora B and has a good inhibitory effect on both. The targeting of the finished drug is improved. The product is highly safe and has little toxic side effects. It can be used in the preparation of drugs related to breast cancer, lung cancer, leukemia, thyroid cancer, prostate cancer, glioma, ovarian cancer, lymphoma, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a structural diagram of the compound 2-(4-carbamoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine. DETAILED DESCRIPTION

[0025] The technical solution of the present invention is further explained below in conjunction with specific implementation methods, but the scope of protection required is not limited to the description.

[0026] Embodiment 1:

[0027] The kinase inhibitory activity was detected by TR-FRET method. Kinase buffer (50 mM HEPES, pH = 7.5, 10 mM MgCl 2 , 2mM DTT, 0.01% Tween-20 and 0.01% BSA) to dilute each kinase protein to a final concentration of 0.1nM. The compound was initially prepared as a 10mM stock solution in DMSO and diluted to 100 times its final concentration with a PBS solution. 200nL of solution 1 and the positive drug Defactinib were added to a 384-well plate, and 10μL of kinase solution was added to each well and incubated at room temperature for 10min. Then, 10μL of a substrate solution containing fluorescein-Poly GT (0.01μM) and ATP (10μM) was added to each well, and the mixture was incubated at 25°C for 30min. Finally, 20μL of a detection solution containing Tb-PY20 antibody (1nM) and EDTA (20mM) was added to the kinase buffer to terminate the reaction. The raw data were collected by a multifunctional microplate reader (excitation wavelength: 340 nm; emission wavelength: 520 nm and 495 nm) and the inhibition rate or IC was calculated. 50 value.

[0028] The inhibitory activity of the compound of the present application, 2-(4-carboxamido)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine, on human breast cancer cells (MDA-MB-231, MCF-7), human chronic myeloid leukemia cells (K562), human thyroid papillary carcinoma cells (TPC-1), human prostate cancer cells (PC-3), human glioma cells (U251), ovarian cancer cells (A2780), human diffuse large B-cell lymphoma (OCI-LY19), and human normal embryonic kidney cells (HEK293) was detected by MTT method, as follows: tumor cells with good growth status and in logarithmic growth phase were taken, and 5×10 3 The cells were seeded in a 96-well plate at a cell density of 100 μL / well and incubated in 5% CO 2After culturing in a cell culture incubator at 37°C for 24 hours, the cells were divided into a blank control group, different concentration groups of the test compound, and a positive drug (Defactinib) group, with 3 replicates in each group. 100 μL of complete medium was added to the blank control group, 100 μL of complete medium containing the corresponding drug concentration was added to the different concentration groups of the test compound, and complete medium containing the corresponding solvent concentration was added to the different concentration DMSO groups. After culturing for 24 hours, 20 μL of MTT was added and stained for 4 hours. The absorbance value was measured under a microplate reader with a wavelength of 570 nm, and the cell survival rate was calculated based on the absorbance value, as follows:

[0029]

[0030] The experimental results are shown in the following table:

[0031] Table 1 Activity of 2-(4-carboxamido)anilino-4-(1H-indol-4-yl)-5-trifluoromethylpyrimidine (1 μmol / L) against different kinases

[0032] Kinase name Inhibition rate (%) Kinase name Inhibition rate JAK1 65 Aurora 91 JAK2 70 AuroraB 97 JAK3 91 Aurora C 82 MAPK1 68 TRKA(G667C) 81 MAPK2 80 AMPKα1 82 MAPK3 68

[0033] Table 2 IC of 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-yl)-5-trifluoromethylpyrimidine inhibiting different kinases and tumor cells 50 value

[0034] Kinase name <![CDATA[IC 50 Value (nmol / L)]]> Tumor cell name <![CDATA[IC 50 Value (μmol / L)]]> JAK3 10.0 MCF-7 3.3 AuroraB 0.5 MDA-MB-231 0.6 FAK 51.0 A549 3.5 K562 0.8 TPC-1 2.1 PC-3 0.9 U251 5.2 A2780 3.6 OCI-LY19 7.2 HEK293 >20

[0035] Table 3 IC of Defactinib inhibiting different kinases and tumor cells 50 value

[0036]

[0037]

[0038] The kinase activity analysis of compound 1 in Table 1 shows that, except for FAK, compound 1 has a very significant inhibitory effect on multiple kinases, and the inhibition rate of 100nM can exceed 60%. Further testing can find (Table 2) that compound 1 has strong selectivity for JAK3 and Aurora B, IC 50 The values ​​are 5 and 100 times higher than those of FAK, respectively, indicating that compound 1 can be used as a good selective inhibitor of JAK 3 and Aurora B, and this activity has not been found in other FAK inhibitors.

[0039] Further testing of anti-tumor activity revealed that compound 1 can effectively inhibit breast cancer cells, lung cancer cells, leukemia cells, thyroid cancer cells, prostate cancer cells, glioma cells, ovarian cancer cells and lymphoma cells, and its activity is significantly stronger than that of the FAK inhibitor Defactinib. The activity of compound 1 in inhibiting MDA-MB-231 breast cancer cells is more than 10 times that of Defactinib. In addition, no toxicity to normal embryonic kidney cells was observed at 20 μM, while Defactinib's IC 50 It is 3.4μM.

[0040] The above data all indicate that compound 1 has great potential in the treatment of breast cancer, lung cancer, leukemia, thyroid cancer, prostate cancer, glioma, ovarian cancer, and lymphoma.

[0041] Finally, it should be pointed out that the above embodiments are only representative examples of the present invention. Obviously, the technical solution of the present invention is not limited to the above embodiments, and there are many variations. All variations that can be directly derived or associated with the content disclosed by ordinary technicians in this field should be considered as the protection scope of the present invention.

Claims

1. Use of a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of an activity inhibitor of Aurora kinase AuroraA, Aurora B and Aurora C, characterized in that: The structural formula of the compound is shown in Formula I below:

2. Use of a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of an activity inhibitor of protein tyrosine kinases JAK 1, JAK 2 and JAK 3, characterized in that: The structural formula of the compound is shown in Formula I below:

3. Use of a compound 2-(4-carboxamido)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of an activity inhibitor of adenylate-activated protein kinase AMPKα1, tropomyosin receptor kinase TRKA, and mitogen-activated protein kinase (MAPK1, MAPK2, MAPK3), characterized in that: The structural formula of the compound is shown in Formula I below: Formula I.

4. Use of a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of breast cancer drugs, characterized in that: The structural formula of the compound is shown in Formula I below:

5. Use of a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of lung cancer drugs, characterized in that: The structural formula of the compound is shown in Formula I below:

6. Use of a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of leukemia drugs, characterized in that: The structural formula of the compound is shown in Formula I below:

7. The use according to claim 6, characterized in that The leukemia is chronic myeloid leukemia.

8. Use of a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of thyroid cancer drugs, characterized in that: The structural formula of the compound is shown in Formula I below:

9. Use of a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of a prostate cancer drug, characterized in that: The structural formula of the compound is shown in Formula I below:

10. Use of a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of a glioma drug, characterized in that: The structural formula of the compound is shown in Formula I below:

11. Use of a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of ovarian cancer drugs, characterized in that: The structural formula of the compound is shown in Formula I below:

12. Use of a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of lymphoma drugs, characterized in that: The structural formula of the compound is shown in Formula I below:

13. Use of a compound 2-(4-carboxamidoyl)anilino-4-(1H-indol-4-oxyl)-5-trifluoromethylpyrimidine in the preparation of a drug for idiopathic pulmonary fibrosis, characterized in that: The structural formula of the compound is shown in Formula I below:

Images