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Application of integrin blocker polypeptide AP25 in preparation of medicines for treating tumor

An integrin blocker, AP25 technology, applied in the field of integrin blocker polypeptide AP25

Inactive Publication Date: 2012-06-13
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are also some integrin blockers that have entered phase II clinical trials, but there are no similar or similar products entering the market in my country

Method used

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  • Application of integrin blocker polypeptide AP25 in preparation of medicines for treating tumor
  • Application of integrin blocker polypeptide AP25 in preparation of medicines for treating tumor
  • Application of integrin blocker polypeptide AP25 in preparation of medicines for treating tumor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Integrin blocker peptide target analysis

[0043] The human umbilical vein endothelial cell (HUVEC) adhesion assay was used to determine the target of the integrin blocker polypeptide. That is, the 96-well plate was first spread with a certain concentration of polypeptide, and the HUVEC cells were incubated with different integrin subunit antibodies, and then the adhesion experiment was carried out. If cells incubated with an antibody show significantly less adhesion to the polypeptide than cells incubated without the antibody, it indicates that the polypeptide interacts with HUVEC through the integrin subunit. The specific operation is as follows: prepare the polypeptide into a 150 μg / ml solution with PBS, spread 100 μl per well on a 96-well plate, place at 4°C for 16 hours, and block with 1% BSA in a 22°C water bath for 4 to 5 hours. HUVEC cells (only use the 2nd to 5th passages) at 37°C, 5% CO 2 Incubate for 4 hours with serum-free endothelial cell culture medium w...

Embodiment 2

[0046] Proliferation Inhibitory Test of Integrin Blocker Peptides on Human Umbilical Vein Endothelial Cells (HUVEC)

[0047] The MTT method was used to detect the activity of the integrin blocker polypeptide in inhibiting the growth of endothelial cells, and the endothelial cells were only used in passages 2-6. HUVEC cells were incubated at 37°C, 5% CO 2 When the incubator was cultivated to a confluence of more than 90%, it was digested with trypsin and collected, and the cells were resuspended in the culture medium and counted under the microscope, and the cell concentration was adjusted to 3×10 4 cells / ml, inoculate the cell suspension into 96-well plate, 100 μl / well, and incubate at 37°C, 5% CO 2 Incubate overnight in the incubator. The integrin blocking agent polypeptide is diluted with culture medium to each predetermined concentration. Endostar was diluted with culture medium to the final concentration. After the cells were completely adhered to the wall, each diluti...

Embodiment 3

[0054] Proliferation Inhibition Test of Integrin Blocker Peptides on Human Gastric Cancer MGC-803 Cells

[0055] MTT method was used to detect the inhibitory activity of integrin blocker polypeptide on the growth of human gastric cancer MGC-803 cells. Tumor cells were incubated at 37°C, 5% CO 2 When the incubator was cultivated to a confluence of more than 90%, it was digested with trypsin and collected, and the cells were resuspended in the culture medium and counted under the microscope, and the cell concentration was adjusted to 2×10 4 cells / ml, inoculate the cell suspension into 96-well plate, 100 μl / well, and incubate at 37°C, 5% CO 2 Incubate overnight in the incubator. The integrin blocking agent polypeptide is diluted with culture medium to each predetermined concentration. Endostar was diluted with culture medium to the final concentration. After the cells were completely adhered to the wall, each dilution was added to a 96-well plate (100 μl / well). Adding the in...

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Abstract

The invention relates to the field of medicines, specifically to an integrin blocker which inhibits generation of tumor vasculature and has integrin endophilicity and binding capacity. The blocker is a polypeptide. The integrin blocker polypeptide can be used to treat solid tumor. The integrin blocker can be applied in the preparation of medicines for treating tumor. The sequence of the integrin blocker is Ala-Cys-Asp-Cys-Arg-Gly-Asp-Cys-Phe-Cys-Gly-Gly-Gly-Gly-Ile-Val-Arg-Arg-Ala-Asp-Arg-Ala-Ala-Val-Pro (the sequence contains two disulfide bonds and the pairing manner is 1-4 and 2-3). The invention is characterized in that the tumor origins from primary or secondary cancer and sarcoma of human head and neck, brain, thyroid, pancreas, lung, liver, oesophagus, stomach, mammary gland, kidney, gallbladder, colon or rectum, ovary, uterus, cervix, prostate, bladder and testis. According to the invention, treatment spectrum of the integrin blocker is greatly expanded, novel thinking and prospect are provided for drug development, and the integrin blocker polypeptide AP25 has substantial social value and market value.

Description

technical field [0001] The present invention relates to the field of medicine, in particular to an integrin blocker polypeptide AP25 capable of inhibiting tumor angiogenesis and having integrin affinity and binding ability. The blocker is a polypeptide, and the integrin blocker polypeptide It can be used in the treatment of solid tumors. Background technique [0002] Malignant tumors are the primary killer of human health. Although traditional tumor therapy has a certain curative effect, due to the non-specific cytotoxicity of chemical drugs, it has relatively large toxic and side effects on patients, and it is also prone to drug resistance, which brings great pain to tumor patients. Since Folkman put forward the theory of tumor angiogenesis in 1972, there have been a large number of reports of treating tumors by inhibiting tumor angiogenesis, and achieved great success. Tumor angiogenesis is the morphological basis of tumor growth and metastasis. The vast majority of mal...

Claims

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Application Information

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IPC IPC(8): A61K38/16A61P35/00
CPCA61K38/16A61P35/00
Inventor 徐寒梅杨永晶胡加亮
Owner CHINA PHARM UNIV
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