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Application and research method of atopaxar hydrobromide as novel JAK-STAT3 signal path inhibitor

A JAK-STAT3, signaling pathway technology, applied in the field of medical applications, can solve problems such as interference with platelet signaling

Active Publication Date: 2019-11-01
QINGDAO MARINE BIOPHARMACEUTICAL RES INST +1
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Problems solved by technology

E5555 is a reversible protease activated receptor (PAR)-1 thrombin receptor antagonist, which can interfere with platelet signal transduction. There is no clear report on the specific target of atopaxabromate

Method used

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  • Application and research method of atopaxar hydrobromide as novel JAK-STAT3 signal path inhibitor

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Embodiment Construction

[0061] For ease of understanding, the technical solutions of the present invention are further specifically described below in conjunction with the accompanying drawings and through embodiments:

[0062] A research method for atopaxa bromide as a novel JAK-STAT3 signaling pathway inhibitor, specifically comprising the following steps:

[0063] S1: Cell culture: cells such as hTERT-BJ, HELA, HepG2, A549, and SKA (the stable transfected strains of the high-throughput screening platform for anti-tumor drugs based on STAT3, which have been successfully constructed in our laboratory) were cultured in DMEM high-glucose culture Base, DU145 and MDA-MB-231 cells were cultured in RPMI-1640 medium at 37°C, 5% CO 2 cultivated under conditions. DMEM high-glucose medium and RPMI-1640 medium contain double anti-penicillin streptomycin and 10% fetal bovine serum FBS.

[0064] S2: Protein extraction: Culture the cells to 80% confluence in a 100mm cell culture dish, wash with 4°C pre-cooled P...

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Abstract

The invention relates to an application of atopaxar hydrobromide as a novel JAK-STAT3 signal channel inhibitor. The atopaxar hydrobromide can be used for inhibiting STAT3 activity and STAT3 downstreamgene expression to control cell proliferation; the atopaxar hydrobromide has an inhibition effect on STAT3 activation induced by cytokines and JAKs self-phosphorylation; in computer molecular simulation, the atopaxar hydrobromide can be combined with an SH2 structural domain of STAT3 and has very high affinity with a JAK kinase JH1 structural domain; and the atopaxar hydrobromide can horizontallyinhibit the tumor cell growth. A research method of the atopaxar hydrobromide as a novel JAK-STAT3 signal pathway inhibitor comprises the following steps: cell culturing; protein extraction; western-Blot; RNA extraction; RT-PCR (Reverse Transcription-Polymerase Chain Reaction) and fluorescent quantitative PCR; luciferase activity analysis; cell viability analysis; cell cycle detection by flow cytometry; cell apoptosis detection by flow cytometry; nude mouse transplantation tumor experiments; computer molecular dynamics simulation docking analysis; and data calculation and processing. The invention lays a foundation for further cell cycle detection and subsequent research and development of novel small molecule drugs.

Description

technical field [0001] The invention belongs to the field of medical application, and relates to a new medical application and research method of atropaxabromic acid, specifically, its application and research method as a JAK-STAT3 signaling pathway inhibitor. Background technique [0002] The JAK-STAT signaling pathway is an important intracellular signal transduction pathway in the process of cell growth, activation, differentiation, apoptosis and its function. Abnormal activation of STAT3 signaling can lead to tumorigenesis, such as breast cancer, lung cancer. Due to their important functions in tumorigenesis, JAKs (Janus kinases) and STATs (signal transducers and activators of transcription) have become popular targets for drug design. Although inhibitors against JAKs and STATs have been approved by FDA, EMA or clinical trials, certain drawbacks such as drug resistance and side effects have also been observed. In addition, as kinase inhibitors, these drugs can bind to ...

Claims

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Application Information

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IPC IPC(8): A61K31/5377A61P35/00C12Q1/66C12Q1/6851G01N15/14G01N21/31G01N33/50
CPCA61K31/5377A61P35/00C12Q1/6851C12Q1/66G01N33/5011G01N21/31G01N15/14C12Q2531/113C12Q2563/107
Inventor 杨金波赵晨阳张心心
Owner QINGDAO MARINE BIOPHARMACEUTICAL RES INST
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