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Method for quickly preparing dasyatis akajei cartilage polypeptide angiogenesis inhibitor

An angiogenesis-inhibiting, polypeptide-based technology, applied in the field of preparation of marine biologically active polypeptides, can solve the problems of low content, large molecular weight of active substances, difficult drug development, etc., and achieve the effect of advanced purification process, significant activity and high accuracy

Active Publication Date: 2015-09-23
ZHEJIANG OCEAN UNIV
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Problems solved by technology

Although it has been reported to prepare angiogenesis inhibitors from cartilage such as red ray and hole ray, the obtained active substances are difficult to develop drugs because of their large molecular weight and low content.

Method used

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  • Method for quickly preparing dasyatis akajei cartilage polypeptide angiogenesis inhibitor
  • Method for quickly preparing dasyatis akajei cartilage polypeptide angiogenesis inhibitor
  • Method for quickly preparing dasyatis akajei cartilage polypeptide angiogenesis inhibitor

Examples

Experimental program
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Embodiment

[0020] Example: A preparation method of red ray cartilage polypeptide angiogenesis inhibitory factor, the preparation process is as follows: red ray cartilage → tissue crushing → guanidine hydrochloride extraction → ultrafiltration → cell membrane stationary phase adsorption → comparative analysis of HPLC fingerprints → angiogenesis inhibitory factor.

[0021] 1) Preparation of cell membrane suspension: Hepatocellular carcinoma cells Bel-7402 in logarithmic growth phase with high expression of vascular endothelial growth factor receptor (VEGFR) were taken, and MEM medium (containing 10% fetal bovine serum) was used to incubate at 37°C, 5% CO 2 Culture in the incubator, when the cell count reaches 10 7 At the same time, the cells were digested with trypsin, centrifuged at 4°C and 1000 r / min for 10 min, the precipitate was washed twice with normal saline, and then added to 50 mM Tris-HCl solution for 30 min sonicated to disrupt the cells. The suspension was centrifuged at 1000...

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Abstract

The invention discloses a method for quickly preparing a dasyatis akajei cartilage polypeptide angiogenesis inhibitor. The method comprises the steps of preparing a cell membrane suspension, preparing a cell membrane stationary phase, preparing and performing enzymolysis on a dasyatis akajei cartilage protein, preparing the dasyatis akajei cartilage polypeptide angiogenesis inhibitor, and the like. The angiogenesis inhibitor can effectively inhibit generation of a blood vessel of chick chorioallantoic membrane (CAM) and has an obvious inhibiting effect to the expression of three pro-angiogenic factors, i.e. vascular endothelial cell growth factor (VEGF), basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF), of a lung cancer tissue of a mouse with Lewis lung cancer.

Description

technical field [0001] The invention relates to a preparation method of marine biologically active polypeptides, in particular to a rapid preparation method of red ray cartilage polypeptide angiogenesis inhibitory factors. Background technique [0002] Both tumor growth and metastasis depend on the formation of new blood vessels. Anti-angiotherapy therapy has the advantages of high efficiency, low toxicity, and less drug resistance. Therefore, searching for angiogenesis inhibitors with significant activity has become a research hotspot of antitumor drugs. Tumor angiogenesis is affected by a variety of factors, and vascular endothelial growth factor (VEGF) is the most important angiogenic factor and the main executor of VEGF signal transduction in endothelial cells, directly participating in the process of neovascularization. Studies have shown that VEGF must play a role by binding to the specific receptor VEGFR-2 located on the cell membrane, so drug screening targeting VEG...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K7/06C07K1/34A61P35/00
Inventor 王斌潘欣迟长凤陈荫赵玉勤孙坤来
Owner ZHEJIANG OCEAN UNIV
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