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Matrix metallo-proteinase inhibitor polypeptide and application thereof

A technology of polypeptide sequence and blood vessel inhibition, which is applied in the field of disease treatment and angiogenesis inhibitors, and can solve problems such as insufficient dosage, irregular medication, persistent and unrelieved symptoms, etc.

Active Publication Date: 2013-06-12
NANJING ANJI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patients often have hand or wrist pain and swelling (especially the swelling of the back of the wrist) as the first symptom, and the symptoms are not relieved. Although common symptomatic treatment can relieve symptoms, it often leads to repeated symptoms due to irregular or insufficient medication

Method used

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  • Matrix metallo-proteinase inhibitor polypeptide and application thereof
  • Matrix metallo-proteinase inhibitor polypeptide and application thereof
  • Matrix metallo-proteinase inhibitor polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Inhibition of several target enzymes by angiogenesis inhibitors.

[0044] Experimental method: recombinant human matrix metalloproteinase-9 [purchased from R&D Systems, USA]. The concentration of matrix metalloproteinase-9 was 92 ng / μL (1 μM). Enzyme activity is detected by cleaving the fluorescently generated peptide substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH 2 [Purchased from R&D Systems, USA] and detected the generated fluorescence values ​​(excitation wavelength = 328nm, detection wavelength = 392nm). All detections were carried out in 100μL reaction system at 37°C.

[0045] The detection of recombinant human matrix metalloproteinase-8 [purchased from Sino Biotechnology Inc., China] is similar to the detection of matrix metalloproteinase-9 and uses the same fluorescent substrate. The reaction was also carried out in a 100 μL reaction system at 37°C. Add 20 μL of recombinant human matrix metalloproteinase-8 (2 ng / μL) to the reaction system during detection. The ...

Embodiment 2

[0053] Migration Inhibition Test of Human Umbilical Vein Endothelial Cells (HUVEC) by Angiogenesis Inhibitor Peptides

[0054] 10 mg / ml Matrigel (BD Company, USA) was diluted 1:2 with HUVEC-specific medium, spread on the transwell chamber membrane, and air-dried at room temperature. The HUVEC cells cultured to the logarithmic growth phase were digested, collected, counted, and the cell concentration was adjusted to 1×10 5 individual / mL. The cells were inoculated into the transwell chamber, 100 μL per well, and each group of test liquid was added into the chamber. Add 0.6 mL of endothelial cell culture medium containing 5% fetal bovine serum and 1% ECGS to the 24-well plate to stimulate cell migration, and place in 5% CO 2 , and cultivated at 37°C for 24h. The culture medium in the well was discarded, the cells were fixed, stained, rinsed with clean water, observed under a microscope and selected four fields of view to take pictures and count. Calculate the migration inhibi...

Embodiment 3

[0062] Angiogenesis Inhibitor Polypeptides Inhibit Growth of Melanoma B16F10C57BL / 6 Transplanted Tumors in Black Mice

[0063] Mice were purchased from Shanghai Slack Experimental Animal Co., Ltd. Cultivate B16F10 cells in vitro, when the cells grow to 90% full, collect the cells with trypsin, and prepare 5×10 6 cells / mL of cell suspension. Inoculate 0.1 mL of cell suspension subcutaneously on the right side of the mouse, and the tumor volume reaches 100 mm in about 7 days 3 . The tumor tissues in the vigorous growth period were taken and ground under sterile conditions, and prepared into 1×10 7 Cell suspension per ml was inoculated subcutaneously in the right axillary of mice with 0.1 mL. Use a vernier caliper to measure the diameter of the transplanted tumor in mice, and wait until the tumor grows to 100-200mm 3 Animals were then randomly grouped. Using the method of measuring the diameter of the tumor, dynamically observe the inhibitory effect of the integrin blocking...

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Abstract

The invention relates to the field of medicines, and in particular relates to an inhibitor of angiogenesis capable of inhibiting matrix metallo-proteinase and migration of vascular endothelial cells. The inhibitor can be used for preventing and treating solid tumor and rheumatoid arthritis. The polypeptide specifically means polypeptide I, polypeptide II, polypeptide III and polypeptide IV (with reference to SeqNO.1, SeqNO.2, SeqNO.3 and SeqNO.4). The polypeptide can be used for treating solid tumor and rheumatoid arthritis. The four inhibitors of angiogenesis can be applied to preparing medicines for treating tumor. The polypeptide is characterized in that tumor is originated from essential or subsequent cancers, melanoma, hemangioma and sarcoma of heat and neck, brain, thyroid, esophagus, pancreas, lung, liver, stomach, breast, kidney, gall bladder, colon or rectum, ovary, cervix uterus, uterus, prostate, bladder and testis.

Description

technical field [0001] The invention relates to the field of medicine, in particular to an angiogenesis inhibitor capable of inhibiting matrix metalloproteinase and inhibiting the migration of vascular endothelial cells, which can be used for the treatment of blood vessel-related diseases including tumors and rheumatoids. Background technique [0002] In recent years, the morbidity and mortality of cancer in my country have been increasing. Unrestricted growth, invasion and metastasis are the hallmarks and characteristics of tumor malignancy and the main causes of treatment failure and death. Therefore, controlling tumor growth, invasion and metastasis is the main measure to improve prognosis and survival rate. In 1971, Folkman first proposed the theory that tumor growth depends on angiogenesis. Tumor angiogenesis is the morphological basis of tumor growth and metastasis. It not only provides nutrients to the tumor, but also exports a large number of tumor cells to the host...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K38/10A61K38/16A61P35/00A61P29/00A61P19/02
Inventor 胡加亮徐寒梅
Owner NANJING ANJI BIOLOGICAL TECH CO LTD
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