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Application of angiogenesis inhibitor polypeptide to preparation of medicine for treating tumor and rheumatoid arthritis

An angiogenesis inhibition and vascular inhibition technology, applied in the field of angiogenesis inhibitors and disease treatment, can solve the problems of insufficient dose, irregular medication, persistent and unrelieved symptoms, etc.

Inactive Publication Date: 2012-10-24
CHINA PHARM UNIV
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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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  • Application of angiogenesis inhibitor polypeptide to preparation of medicine for treating tumor and rheumatoid arthritis
  • Application of angiogenesis inhibitor polypeptide to preparation of medicine for treating tumor and rheumatoid arthritis
  • Application of angiogenesis inhibitor polypeptide to preparation of medicine for treating tumor and rheumatoid arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Preparation and Test of Angiogenesis Inhibitor Polypeptide

[0038] The polypeptide is synthesized by solid-phase synthesis, purified by high-performance liquid chromatography, and the molecular weight of the polypeptide is determined by MS, and the purity of the polypeptide is determined by RP-HPLC.

[0039] Polypeptide I, Peptide II, Peptide III and Peptide IV solid-phase synthesis methods use Fmoc-Rink-resin as the starting material, and then use protected amino acids to inoculate each amino acid in turn. After the peptide incorporation is completed, it is fully washed, and then the peptide is cut and post-treated. The crude product of the angiogenesis inhibitor was obtained. The crude product was dissolved, purified twice by preparative high performance liquid phase, and finally concentrated and freeze-dried to obtain the pure product. The method not only ensures the synthesis efficiency, but also improves the purity.

[0040] 1. The steps of receiving peptide are...

Embodiment 2

[0058] Inhibition of several target enzymes by angiogenesis inhibitors.

[0059] Experimental method: Recombinant human matrix metalloproteinase-9 was expressed by Sf9 insect cells. The enzyme was activated with 0.01 μM matrix metalloproteinase-3 active site, and the buffer used was 100 mM Tris / HCl, pH 7.4, 100 mM NaCl, 10 mM CaCl 2 and 0.01% Tween-20. The concentration of matrix metalloproteinase-9 during activation was 92ng / μl (1μM). Enzyme activity is detected by cleaving the fluorescently generated peptide substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH 2 And detect the generated fluorescence value (excitation wavelength = 328nm, detection wavelength = 392nm). All the detection is carried out in 100μl reaction system at 37°C.

[0060] Recombinant human matrix metalloproteinase-8 is detected similarly to -9 and uses the same fluorogenic substrate. The reaction was also carried out in a 100 μl reaction system at 37°C. Add 20 μl of recombinant human matrix metalloproteinas...

Embodiment 3

[0067] Migration Inhibition Test of Human Umbilical Vein Endothelial Cells (HUVEC) by Angiogenesis Inhibitor Peptides

[0068] 10 mg / ml Matrigel (BD Company, USA) was diluted 1:2 with HUVEC-specific medium, spread on the transwell 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 pieces / 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 0.6ml of endothelial cell culture medium containing 5% fetal bovine serum and 1% ECGS to the 24-well plate to stimulate cell migration, 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. Migration inhibition (MI) was calculated according to...

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Abstract

The invention relates to the field of medicines, in particular to an angiogenesis inhibitor for inhibiting several matrix metal protein enzyme and inhibiting the transfer of the vascular endothelial cell, which can be applied to preparation and treatment of entity tumor and rheumatoid arthritis. The polypeptide comprises a polypeptide I, polypeptide II, polypeptide III and polypeptide IV (Seq NO.1, Seq NO.2, Seq NO.3, Seq NO.4); The polypeptide can be used for treating entity tumor and rheumatoid arthritis. The four angiogenesis inhibitors can be applied to preparation of medicine for treating tumor, and is characterized in that: the tumor is derived from primary of neck, brain, thyroid gland, esophagus, pancreas, lung, liver, stomach, mammary gland, kidney, colon or rectum, ovary, cervix, uterus, prostate, bladder and testis or secondary cancer, melanoma, hemangioma and sarcoma.

Description

technical field [0001] The invention relates to the field of medicine, in particular to an angiogenesis inhibitor capable of inhibiting matrix metalloproteinase and vascular endothelial cell migration, which can be used for the treatment of vascular-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, leading to tumor grow...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K14/00A61K38/10A61K38/16A61K47/48A61P19/02A61P29/00A61P35/00
Inventor 胡加亮邱郑
Owner CHINA PHARM UNIV
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