Method for producing antiangiogenesis agent RT-X in colibacillus and application of antiangiogenesis agent RT-X

Abstract

The invention relates to the technical field of bio-engineering and pharmaceuticals, in particular to a process for producing targeting anti-tumor antiangiogenesis fusion protein RT-X in colibacillus as well as a separation and purification method for the fusion protein RT-X. The fusion protein RT-X is soluble, has biological activity, and comprises RGD and Maspin protein. Through the adoption of the expressed and purified antiangiogenesis agent RT-X with the biological activity, endotheliocyte can be inhibited from migration according to specificity and dose dependence, and an effect for inhibiting angiogenesis is realized in a chicken embryo chorioallantoic membrane animal experiment. The process and the separation and purification method are applicable to research in bio-engineering pharmacy and gene engineering, biochemistry, molecular biology and other fields.

Description

Technical field: [0001] The invention relates to the technical fields of bioengineering and medicine, in particular to a process and a separation and purification method for producing a soluble and biologically active fusion protein RT-X with targeting anti-tumor blood vessel inhibition in Escherichia coli. Background technique: [0002] In recent years, with the in-depth study of tumor anatomy and molecular biology, people have gradually realized that angiogenesis plays a key role in the growth and metastasis of solid tumors. Since Algureza proposed the concept of tumor-induced angiogenesis in 1947, until 1971, folkman clearly proposed the theory that tumor growth and metastasis depend on new angiogenesis. New blood vessels not only provide the necessary nutrients and oxygen for tumor growth, but also help eliminate metabolites , or the way of its distant metastasis (Folkman, J. What is the evidence that tumors are angiogenesis dependent J. Natl. Cancer Inst. 1990: 82: 4-6)...

AI Technical Summary

Problems solved by technology

The αvβ3 antibody currently being researched and developed includes Vitaxin (LM609), which is clinically used for anti-angiogenic therapy (Gutheil, JC., et al.Targeted antiangiogenic therapy for cancer using Vitaxin: a humanized monoclonal antibody to the integrin alphavbeta3.Clin Cancer Res .2000.6, 3056–3061), but the reported effect is not satisfactory (Posey, JA., et al. A pilot trial of vitaxin, a humanized anti-vitronectin receptor (anti alphavbeta3) antibody in patients with metastatic cancer. Cancer Biother Radiopharmac.2001.16,125–132)

Most of the more than 20 integrins recognize extracellular matrix ligands of the RGD (arginine-glycine-aspartic acid) sequence (Dennis MS., et al. Platelet glycoprotein IIb-IIIa protein antagonists from snake venoms:evidence for a family of platelet-aggregation inhibitors. Proc Natl Acad Sci.1990.87, 2471-2475), RGD has the function of antagonizing integrins, reducing the expression of cell surface adhesion molecules and inducing cell apoptosis, but its half-life is short, and it will be destroyed in vivo. Rapid clearance, but the RGD sequence can be used as a carrier, coupled with other active molecules to achieve the effect of targeting tumor vascular endothelium, and has broad application prospects in tumor therapy

[0007] At present, there is no literature report on modifying the sequence of Maspin protein to improve its targeting and anti-tumor activity in vivo, and there is no literature report on the synergistic inhibition of RGD and Maspin protein Angiogenesis

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