A kind of indolinone compound and its preparation method and application

Abstract

The present invention relates to an indolinone compound, which is 1-(4-chlorobenzyl)-3-(hydroxyimino)-4-(4-(trifluoromethyl)phenyl)indoline-2 ‑ketone, which can effectively inhibit the activity of hTERT gene promoter, semi-quantitative PCR experiments and Western blot experiments show that this compound can effectively inhibit the transcription and expression of hTERT, and more importantly, this compound can effectively reduce the length of telomeres in HeLa cells. MTT assay and plate clone assay showed that the compound could inhibit the viability and proliferation of HeLa cells. DAPI staining experiment and flow cytometry analysis showed that the compound could promote cell apoptosis. These results all indicate that the compound can target and inhibit the expression of hTERT and can be used as a new antitumor drug.

Description

technical field

[0001] The invention relates to the field of compounds, in particular to an indolinone compound and its preparation method and application. Background technique

[0002] Telomere (Telomeres) is a small piece of DNA-protein complex that exists at the end of eukaryotic cell linear chromosomes, and it forms a special "cap" structure together with telomere binding proteins. Telomeres exist at the ends of chromosomes. In 1988, the human telomere DNA sequence was confirmed to be a special structure composed of very short and precise tandem TTAGGG repeats and ribonucleoproteins. Telomere ends and telomere-binding proteins form a ring structure instead of a simple linear structure in the cell. The structure and function of telomeric DNA are conserved from single-celled organisms to higher animals and plants. Telomeres provide a protective cap at the ends of chromosomes, and compared to telomeres, the ends of broken chromosomes are more prone to fusion and dissoluti...

Examples

Embodiment

[0038] (1) Screening of hTERT expression inhibitors

[0039]①Cell transfection: HeLa cells were inoculated in two 3cm sterile cell dishes, and the cells were spread to 60-70% and transfected with plasmid pGL2 or pGL2-hTERT promoter respectively, and inoculated into 96-well plates after 6 hours, 8000 per well cells.

[0040] ② Cells treated with small molecule compounds: five concentrations (1, 2, 4, 8, 16 μM) of each compound were used to treat the cells, 3 secondary wells for each concentration, and cultured in an incubator for 36 hours.

[0041] ③Cell lysis: Add 55 μL Extraction buffer [according to the final concentration of 1% TritonX-100, 15mM MgCl2, 4mM EGTA, 1mM DTT, 25mM glycyglycine, make up 10mL with deionized water, store in the dark at 4°C] in the well, and mix well. Lyse on ice for 30 min.

[0042] ④Take 5μL Assay cocktail buffer [add deionized water in order, the final concentration is 30mM ATP (pH 5.0), 0.1M KH2PO4 (pH 7.8), 0.1M MgCl2, mix well, prepare and u...

technical field

[0001] The invention relates to the field of compounds, in particular to an indolinone compound and its preparation method and application. Background technique

[0002] Telomere (Telomeres) is a small piece of DNA-protein complex that exists at the end of eukaryotic cell linear chromosomes, and it forms a special "cap" structure together with telomere binding proteins. Telomeres exist at the ends of chromosomes. In 1988, the human telomere DNA sequence was confirmed to be a special structure composed of very short and precise tandem TTAGGG repeats and ribonucleoproteins. Telomere ends and telomere-binding proteins form a ring structure instead of a simple linear structure in the cell. The structure and function of telomeric DNA are conserved from single-celled organisms to higher animals and plants. Telomeres provide a protective cap at the ends of chromosomes, and compared to telomeres, the ends of broken chromosomes are more prone to fusion and dissoluti...

Examples

Embodiment

[0038] (1) Screening of hTERT expression inhibitors

[0039]①Cell transfection: HeLa cells were inoculated in two 3cm sterile cell dishes, and the cells were spread to 60-70% and transfected with plasmid pGL2 or pGL2-hTERT promoter respectively, and inoculated into 96-well plates after 6 hours, 8000 per well cells.

[0040] ② Cells treated with small molecule compounds: five concentrations (1, 2, 4, 8, 16 μM) of each compound were used to treat the cells, 3 secondary wells for each concentration, and cultured in an incubator for 36 hours.

[0041] ③Cell lysis: Add 55 μL Extraction buffer [according to the final concentration of 1% TritonX-100, 15mM MgCl2, 4mM EGTA, 1mM DTT, 25mM glycyglycine, make up 10mL with deionized water, store in the dark at 4°C] in the well, and mix well. Lyse on ice for 30 min.

[0042] ④Take 5μL Assay cocktail buffer [add deionized water in order, the final concentration is 30mM ATP (pH 5.0), 0.1M KH2PO4 (pH 7.8), 0.1M MgCl2, mix well, prepare and u...

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