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Decapeptide inhibiting angiogenesis and use thereof

A technology of angiogenesis and derivatives, applied in the field of biomedicine

Inactive Publication Date: 2009-12-09
EAST CHINA NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Kisspeptin-10 (Kp-10) is a decapeptide derived from the translation product of human KISS1 gene, which performs biological functions by binding to G protein-coupled receptor-GPCR54 (G protein-coupled receptor-54) At present, there are no reports on the effects of Kp-10 on angiogenesis and tumor growth at home and abroad.

Method used

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  • Decapeptide inhibiting angiogenesis and use thereof
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  • Decapeptide inhibiting angiogenesis and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Kp-10 inhibits the proliferation of human umbilical vein endothelial cells (HUVEC)

[0033] Purpose and principle: MTS Assay was used in the proliferation experiment of human umbilical vein endothelial cells. The MTS Assay is an assay that uses a colorimetric method to indirectly measure the number of viable cells. CellTiter AQueous single-solution reagents consist of a tetrazole compound (inner salt; MTS) and an electron coupling reagent (thiophenazine, PES). MTS can be produced by dehydrogenase in metabolically active cells to form soluble particles in tissue culture medium. The absorbance value of the product measured at 490nm is directly proportional to the number of living cells in the culture.

[0034]Methods: Human umbilical vein endothelial cells (HUVEC) were added to 96-well plate, and the control group and sample addition group were established. The solvent was added to the control group, and different concentrations of Kp-10 polypeptide were ad...

Embodiment 2

[0036] Example 2: Kp-10 inhibits the migration of human umbilical vein endothelial cells in the scratch test

[0037] Purpose and principle: Under the action of vascular endothelial growth factor (VEGF), human umbilical vein endothelial cells can migrate to the scratch area without cells. By observing the amount of endothelial cells migrating into the scratch area, it was evaluated whether KP-10 has the ability to inhibit the migration of human umbilical vein endothelial cells.

[0038] Method: Human umbilical vein endothelial cells were added to the 6-well plate, and when it was about full, a cross-shaped scratch was made with a Tip, the scratched cells were washed with PBS, and culture media containing different drug concentrations and VEGF were added. 5%CO 2 Cultivate in the incubator at 37°C until the cells on both sides of the streak migrate to the streak area.

[0039] Evaluation of the results: Observing the statistics of photographs under an inverted microscope, the ...

experiment example 3

[0040] Experimental Example 3: Kp-10 Peptide Inhibits Boyden Chamber Migration of Human Umbilical Vein Endothelial Cells

[0041] Purpose and principle: The Boyden migration assay is based on the separation of a Boyden chamber into two parts by a microwell. The cells are placed on the upper part, and the lower part contains chemokines. Under the induction of chemokines, the upper layer of cells can migrate through the microporous membrane to the bottom of the membrane and attach to the back of the membrane. This experiment can detect drugs Effects on the ability of endothelial cells to cross microporous membranes.

[0042] Method: Coat the chamber with gelatin, and discard the gelatin after incubating at 37°C in an incubator. Add cell culture medium containing VEGF to the six-well plate outside the small chamber, insert cells into the small chamber, incubate in the incubator for four hours, suck up all the culture medium, fix the small chamber in paraformaldehyde, and wash th...

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Abstract

The invention relates to a decapeptide inhibiting angiogenesis and use thereof, belonging to the technical field of biological medicine. The decapeptide inhibiting angiogenesis provided by the invention is deprived from human Homo sapien with an amino acid sequence of YNWNSFGLRF and the molecular weight of 1303.4, and is named as Kp-10. The invention further provides use of the decapeptide inhibiting angiogenesis for preparing medicines for inhibition of angiogenesis, in particular relevant use of Kp-10 for preparing medicines for inhibition of angiogenesis dependent diseases such as cancer, rheumatoid arthritis, psoriasis, diabetes syndrome, and hemangioma. A series of experimental results in VEGF (Vascular Endothelial Growth Factor) induction shows that Kp-10 has obvious inhibition function in the proliferation experiment of human umbilical vein endothelial cells (HUVEC), transfer experiment, micro-tube forming experiment, angiogenesis experiment of chick embryo allantois and mouse cornea experiment, and the decapetide can be used for preparing medicines for inhibiting angiogenesis.

Description

technical field [0001] The invention relates to a new anti-angiogenesis protein polypeptide Kp-10 and the application of the protein polypeptide, belonging to the technical field of biomedicine. Background technique [0002] Cancer, cardiovascular disease, and diabetes are the three major diseases that threaten human health. Malignant tumors are currently the number one cause of disease death in most countries. At the same time, with changes in the environment and people's living habits, the number of cancer cases is on the rise. [0003] In the 1970s, Judah Folkman of Harvard Medical School first proposed to inhibit tumor growth and metastasis by inhibiting angiogenesis. Tumors can only grow to 2-3 cubic millimeters without the supply of nutrients. If there are new blood vessels connected to the tumor, the tumor can grow rapidly in a geometric progression by obtaining nutrients and oxygen; at the same time, tumor cells can also transfer to other organs through the new blo...

Claims

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Application Information

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IPC IPC(8): C07K7/06A61K38/08A61P35/00A61P19/02A61P27/02A61P27/06A61P27/12A61P9/00A61P17/06A61P19/04A61P9/10A61P35/02
Inventor 易正芳刘明耀
Owner EAST CHINA NORMAL UNIV
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