Application of EGCG structure modified derivatives in preparation of blood vessel regulating and controlling medicine

A technology of derivatives and blood vessels, which is applied in the application field of EGCG structurally modified derivatives in the preparation of blood vessel regulating drugs, can solve the problems that there are no literature reports on the application of EGCG structurally modified derivatives, and achieve the goal of inhibiting the formation of new blood vessels and tumor cells Effects of transfer, regulation of angiogenesis, and convenience of material collection

Inactive Publication Date: 2015-11-11
GUANGXI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] The application of the above-mentioned patents in the treatment of type 2 diabetes mellitus, the application of food and medicine in the prevention and treatment of myocardial energy metabolism disorder, the application of the medicine in the treatment or prevention of enterovirus 71 infection, the application of the medicine in the treatment or prevention of hepatitis C virus infection However, there is no literature report on the application of EGCG structurally modified derivatives in the preparation of vascular regulation drugs.

Method used

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  • Application of EGCG structure modified derivatives in preparation of blood vessel regulating and controlling medicine
  • Application of EGCG structure modified derivatives in preparation of blood vessel regulating and controlling medicine
  • Application of EGCG structure modified derivatives in preparation of blood vessel regulating and controlling medicine

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Experimental program
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Effect test

Embodiment 1

[0031] 1. Real-time fluorescent quantitative PCR (Real-timePCR) method to detect the effect of EGCG structural modification derivatives on the expression of HIF-1α and VEGF genes in liver cancer cells

[0032] 1.1 Culture of human liver cancer cells SMMC-7721

[0033] Well-growing SMMC-7721 liver cancer cells were taken and divided into two groups: normal oxygen control group

[0034] (drug-free medium) and hypoxia group. The hypoxic group was further divided into: the hypoxic control group (without drug-containing medium), the high-dose EGCG group (EGCG-H, 50-100 μmol / L), the medium-dose EGCG group (EGCG-M, 25-50 μmol / L) , EGCG low dose group (EGCG-L, 1~25μmol / L), EGCG ethylated derivative Y1~6 high dose group (25~50μmol / L), EGCG ethylated derivative Y1~6 medium dose group (10 ~25μmol / L), EGCG ethylated derivative Y1~6 low dose group (1~10μmol / L). Cells in each group were cultured in normoxic and hypoxic incubators for 16 hours after adding drugs. Normoxic culture conditi...

Embodiment 2

[0060] 2. Western blot method to detect the effect of EGCG structurally modified derivatives on the expression of HIF-1α and VEGF proteins in liver cancer cells

[0061] 2.1 Cell culture and grouping are the same as 1.1.

[0062] Hepatoma cells were cultured in normoxia and hypoxia for 16 hours after adding the drug, discarded the culture medium, rinsed twice with PBS, added 1ml of PBS to the culture bottle, collected the cells with a cell scraper, transferred to an Eppendorf tube, and centrifuged at 12,000g for 5 minutes. Discard the supernatant, add 500 μl of protein lysate, shake with a vortex shaker, and place on ice for 30 minutes. Centrifuge at 12,000 g for 10 min, collect the supernatant and measure the protein concentration with a BCA kit. SDS-PAGE protein buffer was mixed in proportion, boiled for 5 minutes, aliquoted, and stored at -20°C for later use.

[0063] gel

[0064] Use 8%-12% SDS-PAGE gel, the protein loading amount is 15ug, the stacking gel is 80V30min, ...

Embodiment 3

[0072] 3MTT method to detect the effect of EGCG structurally modified derivatives on the growth of human umbilical vein endothelial cells

[0073] 3.1 Experimental method

[0074] Take HUVEC cells in the logarithmic growth phase with normal morphology, digest, centrifuge, remove the supernatant, and collect the cell pellet. Add an appropriate amount of DMEM high-glucose culture medium and pipette evenly to form a single-cell suspension, and count under a microscope. Adjust the cell suspension density to 1×10 4 / mL, inoculated in 96-well culture plate, inoculated 100 μL per well, placed in 5% CO 2 Cultivate for about 24 hours in an incubator at 37° C. and saturated humidity. EGCG, AcEGCG, and EGCG ethylated derivative Y6 were diluted with DMEM high-glucose culture medium to 5 concentrations as drug groups, and the drug doses of EGCG were 200 μg / ml, 150 μg / ml, 100 μg / ml, 50 μg / ml, and 25 μg / ml, the drug doses of Y1~6 were 120μg / ml, 80μg / ml, 40μg / ml, 20μg / ml, 10μg / ml respect...

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Abstract

The invention relates to application of EGCG structure modified derivatives in preparation of blood vessel regulating and controlling medicine. The EGCG structure modified derivatives have stability and pharmacological activity higher than those of EGCG, and the blood vessel growth regulating and control functions can be achieved by regulating and controlling the expression of VEGF and CBR1 and restraining the growth of vascular endothelial cells.

Description

technical field [0001] The invention relates to pharmacology and medicine, and mainly relates to the application of a class of EGCG structure-modified derivatives in the preparation of blood vessel regulating drugs. Background technique [0002] Angiogenesis plays an important role in the growth and metastasis of tumor cells, the damage of ocular neovascularization to ocular structure and function, diabetic neovascularization, renal vascular proliferative disease, coronary heart disease and other neovascularization-related diseases. Inhibition of new angiogenesis can cut off the nutrient supply of tumors, promote the regression and shrinkage of tumors, reduce the chance of blindness in the eyes, reduce the complications of diabetes, protect the normal physiological functions of the kidneys, and prevent and treat the occurrence and development of coronary heart disease. confirmed by a large number of research results. The growth and metastasis of various tumors are accompani...

Claims

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Application Information

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IPC IPC(8): A61K31/353A61P35/00A61P35/04
Inventor 梁钢李丽陈润丽孙悦文唐安州刘布鸣周焕弟温燕付丽香陈燕燕
Owner GUANGXI MEDICAL UNIVERSITY
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