Application of lamprey crbgp in the preparation of anti-angiogenesis drugs

A CRBGP, lamprey technology, applied in drug combination, cardiovascular system diseases, pharmaceutical formulations, etc., can solve the problems of inhibiting angiogenesis function, and no cysteine-rich secreted protein has yet been found.

Inactive Publication Date: 2016-09-14
LIAONING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, there has been no report on the inhibition of angiogenesis by cysteine-rich secreted proteins

Method used

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  • Application of lamprey crbgp in the preparation of anti-angiogenesis drugs
  • Application of lamprey crbgp in the preparation of anti-angiogenesis drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0066] The present invention uses molecular sieve chromatography (Sephadex G75) to separate and obtain lamprey natural CRBGP with electrophoretic purity from Japanese lamprey oral glands.

[0067] The lamprey CRBGP gene was connected to the pEGX-4T-1 expression vector (or other forms of vector) by gene cloning methods to induce expression in E.coli Rosetta, and the recombinant lamprey CRBGP was purified by GST affinity chromatography, The target protein was detected by 12% polyacrylamide gel electrophoresis and mass spectrometry, and its purity and accuracy were consistent with the target protein.

[0068] experiment:

[0069] 1. The inhibitory effect of lamprey CRBGP on the proliferation of HUVEC cells was determined by MTT assay.

[0070] (1) Seed HUVEC cells in good condition in a 96-well plate and culture them in RPMI 1640 medium containing bFGF (final concentration: 3 ng / mL) for 24 hours;

[0071] (2) Lamprey natural CRBGP with different concentration gradients (5 μL, ...

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Abstract

The invention discloses application of lamprey CRBGP in preparation of anti-angiogenesis medicines. The application disclosed by the invention proves that natural and recombinant CRBGP of lamprey can inhibit proliferation of bFGF-induced human umbilical vein endothelial cells (HUVEC) and has dose dependence, and the IC50 value is 4.8mu mol / L. Meanwhile, the natural and recombinant CRBGP of lamprey has an effect of inhibiting angiogenesis of bFGF-induced six-day-age chick chorioallantoic membrane (CAM) and can be applied to preparing low-toxicity high-efficiency anti-angiogenesis medicines.

Description

technical field [0001] The invention relates to a use of lamprey CRBGP, in particular to an application of lamprey CRBGP in preparing anti-angiogenesis drugs. Background technique [0002] Angiogenesis, the development of new microvessels from pre-existing vessels, plays an important role in some normal physiological processes such as embryonic development, wound healing, and corpus luteum formation. However, unexpected angiogenesis can also lead to a variety of diseases, such as chronic inflammation, diabetic retinopathy, choroidal neovascularization, psoriasis, myelodysplastic syndrome, rheumatoid arthritis, and tumors, which seriously harm Human health and quality of life. Therefore, the development of low-toxicity and high-efficiency anti-angiogenesis drugs is currently a hot spot for the treatment of abnormal angiogenesis diseases related to inflammation. [0003] Cysteine ​​rich secretory proteins (CRISP) are a class of proteins that are highly conserved in evolution...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/17A61P9/00
Inventor 李庆伟肖蓉刘宇姜琪王红艳
Owner LIAONING NORMAL UNIVERSITY
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