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Bombesin-oriented anti-tumor polypeptide and preparation method and application thereof

An anti-tumor, bombesin technology, applied in the application field of medicine

Inactive Publication Date: 2011-05-18
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is also no report on the use of bombesin and its homologues and macromolecular proteins / peptides to construct fusion peptides for the treatment of hematological tumors

Method used

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  • Bombesin-oriented anti-tumor polypeptide and preparation method and application thereof
  • Bombesin-oriented anti-tumor polypeptide and preparation method and application thereof
  • Bombesin-oriented anti-tumor polypeptide and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1 Preparation of antitumor peptide of the present invention

[0049] Unless otherwise specified, all cell lines of the present invention were purchased from the American ATCC cell bank. The cell lines used in the experiment include: human umbilical vein endothelial cells (HUVECs), vascular endothelial cells (ECV304), human embryonic lung fibroblasts (MRC-5), human prostate cancer cells (Du145), human breast cancer cells (MCF -7), human chronic myeloid progenitor cells (K562), human Burkitt's B lymphoma cells (Raji), human acute T lymphoma cells (CEM), human acute T lymphoblastic leukemia cells (Molt4), human acute leukemia early childhood granulocytes (NB4); and freshly isolated human decidual stromal cells (HDSCs). The peripheral blood lymphocytes of normal people were obtained from laboratory volunteers, and the peripheral blood lymphocytes of patients with acute myeloid leukemia were separated by gradient centrifugation using low-concentration Ficoll mono...

Embodiment 2 B28

[0057] In vitro anti-tumor experiment of embodiment 2 BB28

[0058] The fusion peptide obtained in Example 1 was used to prepare peptide solutions of 2.5, 5, 7.5, 10, 12.5, and 15 μM, and the following experiments were carried out:

[0059] 1) Cytotoxicity test method

[0060] Adherent cells were seeded in 96-well plates at 10,000 cells / well and cultured overnight to allow them to adhere to the wall. On the next day, the supernatant medium was discarded and replaced with 100 μl medium containing 2% bovine serum albumin (BSA), and peptides of different concentrations were added for reaction. Non-adherent cells were suspended in 2% BSA medium, and after 10,000 cells / well were inserted into a 96-well plate, different concentrations of peptides were directly added. After the peptide acts on the cells, the CCK-8 kit can also be used to quantitatively determine the number of surviving cells by detecting the activity of mitochondrial enzymes in the cells. The specific operation is...

Embodiment 3 B28

[0079] Example 3 BB28 anti-tumor experiment in vivo

[0080] K562 cells (1×10 7 ) suspended in 100 μl of normal saline, and subcutaneously injected into 6-8 week old nude mice to establish a tumor model.

[0081] 1) Anti-tumor effect of intraperitoneal injection of BB28

[0082] When the tumor grows to 0.03-0.05mm 3 8 experimental animals were randomly divided into 2 groups. Animals in the experimental group were intraperitoneally injected with BB28 at a concentration of 20 mg / kg for 5 consecutive days. The control group was given the same volume of phosphate buffer. The diameter of the tumor was measured with a vernier caliper every day, and the volume of the tumor was calculated according to the formula (length x width x width x 0.5). After the experiment, the experimental animals were sacrificed, and the tumor tissues were taken out, photographed and weighed.

[0083] The result is as Figure 10 As shown in A, intraperitoneal injection of BB28 can significantly inhib...

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Abstract

The invention discloses a bombesin-oriented anti-tumor polypeptide, which is a fusion peptide formed by connecting bombesin and cationic antibacterial peptide from bovine bone marrow cells, wherein the cationic antibacterial peptide has an active area of damaged mitochondria. The anti-tumor polypeptide has obvious targeted inhibiting effect on solid tumors and blood tumors, improves the safety of medicaments, provides a new path for clinical medicament application, and has good application prospect.

Description

technical field [0001] The invention relates to a bombesin-directed anti-tumor polypeptide, especially an anti-tumor polypeptide in which bombesin is linked with an antibacterial peptide from bovine bone marrow cells, and its application in the preparation of drugs against various solid tumors and hematological tumors . Background technique [0002] Cancer is one of the major diseases that threaten human health. The biggest limitation of existing cancer treatment drugs is the lack of selectivity, which kills normal cells while eliminating cancer cells. Using tumor cell-specific recognition molecules as carriers to carry cytotoxic drugs (radiation and chemotherapy drugs, protein / peptide toxins, etc.) is one of the important ways to achieve targeted tumor therapy. Studies have proved that antibodies and their functional fragments that can specifically recognize tumor cell surface antigens, and cytokines that can specifically recognize tumor cell surface receptors are good dru...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00A61K38/17A61K38/10A61P35/00A61P35/02
Inventor 卢晓风蔡华伟杨浩万琳刘珊程惊秋李幼平李胜富张杰
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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