Liver targeted peptide-recombinant human endostatin fusion protein, and preparation method and application thereof
一种内皮抑素、融合蛋白的技术,应用在医药生物领域,能够解决毒副作用、血管形成不良影响等问题,达到提高特异性、减少全身用量、提高量效比的效果
Active Publication Date: 2013-09-25
GUANGDONG PHARMA UNIV
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Problems solved by technology
However, during treatment, Endostar is easy to diffuse and degrade in the body, resulting in a relatively uniform tissue distribution and failing to form an effective anti-tumor concentration at the lesion site. In order to achieve clinical effects, the only way to increase the dose of the drug is; however, increasing the dose will cause more severe Toxic and side effects, such as impaired cardiac function, diarrhea, etc., especially potential adverse effects on angiogenesis in physiological states such as embryogenesis and wound healing
Method used
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Experimental program
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Embodiment 1
[0038] S1. Cloning of the ES gene
[0039] Cell culture and total RNA extraction: Embryonic liver cells L-02 were cultured in 1640 cell culture medium containing double antibodies (100U / ml penicillin, 100ug / ml streptomycin) and 10% fetal bovine serum until the cells were confluent. TIANGEN TRNzol Total RNA Extraction Kit Instructions Operation, every 10cm 2 Add 1ml TRNzol to the area to extract total cellular RNA.
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Abstract
The invention discloses a liver targeted peptide-recombinant human endostatin fusion protein, and a preparation method and application thereof. According to the invention, 19 amino acids in the N terminal conserved I region of plasmodium circumsporozoite protein (CSP) capable of being specifically combined with hepatocyte surface receptor--heparan sulfate proteoglycan (HSPG) are fused to the carboxyl terminal of a novel human endostatin (nhES), the amino terminal of which is added with 9 amino acid sequences (MGGSHHHHH), by a gene engineering method, thereby obtaining the liver targeted peptide-recombinant human endostatin fusion protein (ES-CPS). The fusion protein is easy to purify, can be specifically used in livers for inhibiting neovascularization in a targeted mode, enhances the local concentration of the medicine in the foci, reduces the consumption to the whole body, and lowers the toxic or side effect. The invention provides an idea and scientific reference for clinically developing medicines for treating liver cancers by targeted inhibition of neovascularization, and has huge scientific meanings and practical value for further enhancing the liver cancer treatment level in China.
Description
technical field [0001] The invention belongs to the field of medical biotechnology, and more specifically relates to a fusion protein of liver-targeting peptide and recombinant human endostatin, its preparation method and application. Background technique [0002] Primary liver cancer (HCC) is one of the main malignant tumors in my country, with an annual incidence rate of 10-150 / 100,000 people, second only to lung cancer and gastric cancer, and is the third malignant tumor in my country, with an annual mortality rate of about 34.6 per 100,000 people, second only to lung cancer. Primary liver cancer is also the leading cause of death in patients with cirrhosis. It is also a common disease in the world. Primary liver cancer ranks fifth in the world in terms of incidence of malignant tumors, and ranks third in mortality. At present, there are about 350,000 new cases of primary liver cancer in my country every year, accounting for 55% of the new cases in the world. About 320,...
Claims
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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/70C12N1/21A61K38/17A61K47/48A61P35/00C12R1/19
CPCY02A50/30
Inventor 朱家勇马艳金小宝卢雪梅
Owner GUANGDONG PHARMA UNIV
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