Dasyatis akajei cartilage polypeptide angiogenesis inhibitor

An angiogenesis inhibition and peptide technology, applied in the direction of peptides, can solve the problems of low content, difficult drug development, and large molecular weight of active substances, and achieve the effect of advanced purification process, high accuracy, and significant activity

Active Publication Date: 2015-09-02
ZHEJIANG OCEAN UNIV
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  • Claims
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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

Method used

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  • Dasyatis akajei cartilage polypeptide angiogenesis inhibitor
  • Dasyatis akajei cartilage polypeptide angiogenesis inhibitor
  • Dasyatis akajei cartilage polypeptide angiogenesis inhibitor

Examples

Experimental program
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Embodiment

[0020] 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, HPLC fingerprint comparison analysis, 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 r / min, 4°C for 5 mi...

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Abstract

The invention relates to a dasyatis akajei cartilage polypeptide angiogenesis inhibitor, and a rapid preparation method and an application thereof. An amino acid sequence of the angiogenesis inhibitor is Gly-Tyr-Ile-Cys-Pro-Gln-Trp; and the molecular weight of ESI-MS (electrospray ionization mass spectrometry) detection is 865.98Da. The angiogenesis inhibitor is capable of effectively inhibiting angiogenesis of chick chorioallantoic membrane (CAM), and has an obvious inhibiting effect on expression of three pro-angiogenic factors, namely a Lewis lung cancer mice lung cancer tissue-loaded vascular endothelial cell growth factor (VEGF), a basic fibroblast growth factor (bFGF) and a platelet-derived growth factor (PDGF).

Description

technical field [0001] The invention relates to a marine biological active polypeptide, in particular to a red ray cartilage polypeptide angiogenesis inhibitory factor. 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 VEGFR-2 has become an important strategy for anti-tumor a...

Claims

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Application Information

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IPC IPC(8): C07K7/06
Inventor 王斌潘欣迟长凤陈荫赵玉勤孙坤来
Owner ZHEJIANG OCEAN UNIV
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