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Synthesized polypeptide and application

A technique for synthesizing peptides, -his-, applied to medical preparations containing active ingredients, peptides, desipeptides, etc., can solve the problems of low bioavailability, tiny TIMPs, difficult to extract, etc., and achieve broad application prospects and therapeutic effects Good results

Inactive Publication Date: 2007-02-21
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the content of TIMPs in tissues is extremely small and difficult to extract; artificially recombined TIMPs are macromolecular proteins, and there are problems such as low bioavailability

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Synthesis, separation, purification and disulfide bond construction of BSC-1, BSC-2 and BSC-3:

[0051] Synthesized by Fmoc / tBu solid-phase peptide synthesis method (SPPS). The synthesized solid phase support is 2-chlororityl resin, each step of the coupling reaction is one, and the Kaiser test is used to detect the reaction process. After synthesizing the desired peptide chain, use Reagent B (TFA 88%, phenol 5%, H 2 O 5%, TIS 2%) cut off the polypeptide from 2-chlororityl resin, and precipitated with cold ether to obtain the crude polypeptide. The disulfide bonds of the crude product were reduced with mercaptoethanol, then dissolved in DMSO to form a dilute solution, and the intramolecular disulfide bonds were formed by agitating with air overnight.

[0052] RT-HPLC purified polypeptide, C18 column (10mm×200mm, 300 Ȧ particle size), linear gradient elution H 2 O / acetonitrile 0.1% TFA. Freeze-dried to obtain 12mg of SN-1, 13mg of SN-2, and 10mg of SN-3. The...

Embodiment 2

[0053] Example 2 Affinity experiment between substrate and receptor CXCR4

[0054] Cultivate SN-1, 2, 3 and CXCR4+ U373-MAGI-CXCR4 CEM cells, add fluorescein-labeled CXCR4 monoclonal antibody 12G5, incubate for a certain period of time, and measure the fluorescence intensity of the cells. It shows that the polypeptide substrate has a strong affinity with the receptor CXCR4. The IC50 of the sequences BSC-1, BSC-2 and BSC-3 are shown in Table (1):

[0055]

Embodiment 3

[0056] Example 3 The substrate inhibits the interaction between SDF-1α and CXCR4

[0057] Culture fluorescently labeled SDF-1α and CXCR4+ U373-MAGI-CXCR4 CEM cells, add the substrate to be tested, incubate for a certain period of time, and measure the fluorescence intensity of the cells. It shows that the strong affinity between the polypeptide substrate and the receptor CXCR4 inhibits the interaction between the natural ligand SDF-1α and CXCR4, and the IC50 of the sequences BSC-1, BSC-2 and BSC-3 are shown in Table (1).

[0058] The results indicated that the peptide substrate may target the interaction of SDF-1α / CXCR4.

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PUM

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Abstract

A synthetic polypeptide and its amino acid sequence are disclosed. It can be used to prepare the medicines for preventing and treating tumor transfer.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a synthetic polypeptide. Background technique [0002] Tumor invasion and metastasis are the most essential differences between malignant and benign tumors, and they are also one of the important reasons for the failure of treatment in patients with malignant tumors. Tumor invasion refers to the process in which tumor cells in the primary lesion destroy the natural barrier formed by the extracellular matrix and basement membrane, invade adjacent normal tissues, and cause necrosis and disease. The process of forming metastases in organs distant from the primary site. Clinically, about 30% of newly diagnosed solid tumor patients have visible metastases, especially some highly metastatic tumors such as breast cancer, prostate cancer, ovarian cancer and melanoma. Although the current comprehensive treatment based on surgery can achieve good curative effect on early tumors, the inv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/00A61K38/16A61P35/00
Inventor 卜宪章邵伟艳古练权曹颖男于志文
Owner SUN YAT SEN UNIV