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Simulation oligopeptide of vascular endothelial growth factor (VEGF) epitope and application of simulation oligopeptide

A growth factor and vascular endothelial technology, applied in the direction of peptides, anti-tumor drugs, DNA/RNA fragments, etc., can solve the problems that limit the wide application of peptide drugs, achieve obvious anti-angiogenic effect, obvious anti-tumor, easy to synthesize and construct Effect

Inactive Publication Date: 2014-08-27
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, macromolecular peptides may also trigger the body's immune response, which greatly limits the wide application of peptide drugs

Method used

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  • Simulation oligopeptide of vascular endothelial growth factor (VEGF) epitope and application of simulation oligopeptide
  • Simulation oligopeptide of vascular endothelial growth factor (VEGF) epitope and application of simulation oligopeptide
  • Simulation oligopeptide of vascular endothelial growth factor (VEGF) epitope and application of simulation oligopeptide

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Experimental program
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Effect test

Embodiment 1

[0023] Through the method of phage display, the analog short peptide CPU-510-02 of endothelial cell growth factor VEGF epitope was obtained:

[0024] (1) Obtain the nucleotide sequence of the analog short peptide CPU-510-02 of endothelial cell growth factor VEGF antigen epitope through the screening of phage random peptide library:

[0025] A. Biopanning of phage random 7-peptide library: Anti-VEGF monoclonal antibody was coated with coating solution (0.1M NaHCO 3 , pH8.6) was diluted to 100 μg / ml, and 150 μl of the solution was used to coat one well of the microtiter plate, overnight at 4°C. Aspirate the coating solution, add blocking solution (0.1M NaHCO 3, pH8.6, 5mg / ml BSA), at 4°C for at least one hour. Aspirate the blocking solution and wash 6 times with 0.1% TBST [TBS+0.1% (v / v) Tween-20]. Dilute 10 μl of the original phage polypeptide library with 100 μl TBST, then add it to the wells coated with anti-VEGF monoclonal antibody, react at room temperature for 1 hour, p...

Embodiment 2

[0033] Proliferation inhibitory test of peptides on human umbilical vein endothelial cells HUVEC, melanoma cells B16F10, gastric cancer cells SGC7901 and liver cancer cells HepG2:

[0034] (1) MTT method to measure the inhibitory effect of epitope peptides on HUVEC proliferation in the presence of VEGF: take human umbilical vein endothelial cells in logarithmic growth phase HUVEC, digest with an appropriate amount of 0.25% trypsin solution, and shake gently to make the digestive juice evenly act For the cells, remove the digestive juice when the sheets of cells are round and shrunk to a single cell state, quickly add an appropriate amount of medium, and blow gently with a pipette to form a single-cell suspension; take the single-cell suspension and count it as 5000 cells per well Inoculated in a 96-well plate (edge ​​wells were filled with sterile PBS), with DMEM containing 10% calf serum at 37 ° C, containing 5% CO 2 Cultured in a constant temperature incubator for 24 hours t...

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Abstract

The invention discloses simulation oligopeptide CPU-510-02 of the vascular endothelial growth factor (VEGF) epitope and an application of the simulation oligopeptide. The amino acid sequence of the simulation oligopeptide CPU-510-02 is PSCVPLM, and the nucleotide sequence of the simulation oligopeptide CPU-510-02 is CCGTCCTGCGTTCCGCTGATG. An in-vitro experiment shows that the simulation oligopeptide has an obvious inhibition effect in a vascular endothelial cell multiplication process and a tumor cell multiplication process; and therefore, the simulation oligopeptide can be used for preparing polypeptide vaccines, tumor vessel growth inhibitors or tumor guide medicines and has great reference signification for diagnosing generation and development of tumors by taking the VEGF acceptor as a target, discussing the development of tumor vessel growth inhibitors by micro-molecule polypeptides and developing and researching tumor guide medicines.

Description

technical field [0001] The invention belongs to a vascular endothelial cell growth factor VEGF antigen, in particular to a short analog peptide of the vascular endothelial cell growth factor VEGF antigen epitope and application thereof. Background technique [0002] VEGF is a homodimeric protein with a relative molecular weight of 34-42KD that was first purified by Ferrara in the in vitro culture medium of bovine pituitary follicular astrocytes in 1989. The human VEGF gene structure is located on chromosome 6p21.3, consisting of 8 exons and 7 introns alternately. According to the splicing mode of its mRNA, five different VEGF isoforms can be produced, including VEGF206, VEGF189, VEGF165, VEGF145 and VEGF121. These five isoforms of VEGF all have the activity of inducing endothelial cell proliferation and are effective angiogenesis and permeability inducing factors. and ability. [0003] VEGF acts by binding to its corresponding VEGF receptor. Currently cloned VEGF recepto...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C12N15/11A61K38/08A61P35/00
Inventor 刘煜王刚甘牡丹王登徐剑王旋宋梦丹
Owner CHINA PHARM UNIV
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