Integrin blocker polypeptide as well as preparation method and application thereof

An integrin blocker and technology for use, applied in the direction of peptide preparation methods, chemical instruments and methods, peptides, etc., can solve the problems of short half-life, easy degradation in the body, etc., achieve small side effects, reduce the number of amino acids, and reduce synthesis cost effect

Inactive Publication Date: 2014-09-24
NANJING ANJI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Aiming at the problem that the existing HM-3 has a short half-life and is easy to degrade in vivo, the present invention provides an integrin blocker polypeptide and a preparation method thereof, an integrin blocker polypeptide c

Method used

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  • Integrin blocker polypeptide as well as preparation method and application thereof
  • Integrin blocker polypeptide as well as preparation method and application thereof
  • Integrin blocker polypeptide as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Preparation and Test of Integrin Blocker Polypeptide Synthetic Peptide Ⅰ-X

[0057] The peptide solid-phase synthesis method is adopted, which uses wang resin (Wang resin) as the starting material, and then uses the protected amino acid Fmoc-arginine-OH to sequentially connect one peptide to eight, nine, ten to seventeen peptides, and the peptide connection work is completed After that, it is fully washed, and then the peptide is cut and post-treated to obtain the crude peptide. Purify the crude product by first dissolving it, then using preparative high-performance liquid phase for two purifications, and finally concentrating and freeze-drying to obtain the pure product. Specific steps are as follows:

[0058] 1. Synthesis:

[0059] Weigh 13g of Wang resin, pour it into a 1L reaction column with glass sand core, and add 130ml of CH2Cl2 to fully expand the resin.

[0060] Uncapping: Add 25mL of hexahydropyridine / DMF decapping solution, seal it and place it in a shake...

Embodiment 2

[0092] Protective effect of integrin blocker polypeptide on in vivo immune protection of adjuvanted rat arthritis animal model

[0093] An animal model of adjuvant arthritis in rats was constructed to study the therapeutic effect of integrin blocker polypeptide on adjuvant arthritis (AA) rats. Rats were used as the test animals. 238 clean-grade wistar rats, male, with a body weight of 150g-180g, were randomly divided into 34 groups, which were blank control group, model control group, and three dosage groups of polypeptide Ⅰ-X low, middle and high ( 3, 6, 9mg / kg) and the positive drug control group (methotrexate 1mg / kg), plus a HM-3 treatment dose group for comparison. Except for the blank group, rat AA models were established in each experimental group on the 0th day by injecting complete Freund's adjuvant containing inactivated Mycobacterium tuberculosis (H37RA, 10mg / ml) in the left hindquarters of the rats 0.08ml caused rat adjuvant arthritis model. Secondary inflammation...

Embodiment 3

[0112] Effects of Integrin Blocker Peptide on Corneal Neovascularization in Mice

[0113] 1) Preparation of corneal neovascularization model induced by alkali burn in BALB / c mice:

[0114] Fifty-five healthy BALB / c mice, male, weighing 20-25 g, were examined under a slit-lamp microscope for the anterior segments and appendages of both eyes to exclude eye lesions. 0.3% ofloxacin eye drops were instilled 1 day before the preparation of the alkali burn model, twice a day. After mice were anesthetized by intraperitoneal injection of 1.8% Avertin, a single-layer filter paper sheet with a diameter of 2 mm was clamped with tweezers, immersed in 1mol / L sodium hydroxide solution to make it saturated, excess liquid was removed, and the filter paper sheet was placed in BALB / c mouse corneal central 40S, discard the filter paper, immediately rinse the burn area and conjunctival sac with 15mL of PBS for 1min. Wipe off excess water with a cotton swab, and rotate the corneal spatula paralle...

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Abstract

The invention discloses an integrin blocker polypeptide as well as a preparation method and application thereof, and belongs to the field of biopharmacy. The integrin blocker polypeptide only has 9 to 17 amino acids from the polypeptide I to the polypeptide X, and is synthesized by using a solid-phase synthesis method, so that the integrin blocker polypeptide also has an in-vivo immune protection function and can be used for treating arthritis even the number of the amino acids in the integrin blocker polypeptide is reduced. The integrin blocker polypeptide has the function of inhibiting neovascularization and has the activity of inhibiting endothelial cell migration so as to prevent and treat diseases relevant to blood vessels and inflammations comprising oculopathy relevant to neovascularization. The integrin blocker polypeptide has the high anti-tumor activity to various types of tumors, so that the cost is greatly lowered. The integrin blocker polypeptide has the good function for treating chronic infectious arthritis, so that the adaptability and the applicable diseases of the integrin blocker polypeptide are increased. Thus, the social and economic values of the integrin blocker polypeptide are enlarged.

Description

technical field [0001] The invention relates to the field of biotechnology medicine, in particular to the application of the integrin blocker polypeptide and its preparation method and preparation treatment for the prevention and treatment of arthritis, tumor treatment and ophthalmic diseases. Background technique [0002] Arthritis, inflammation caused by bacteria, tumors, eye diseases such as AMD are all called vascular-related diseases. Rheumatoid arthritis (RA) is the most common clinical inflammatory joint disease and one of the main disabling factors. In the pathological process of RA, angiogenesis is a landmark histological change. The formation of new blood vessels is accompanied by synovial hyperplasia and inflammatory cell infiltration, which is the basis of pannus formation and joint destruction in RA. There should be no blood vessels in the articular cartilage due to some abnormal changes, and new blood vessels are formed, which makes the cartilage erode, causin...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K7/06C07K1/16C07K1/06C07K1/04A61P19/02A61P9/10A61P27/02A61P35/00
Inventor 康梦实张力君
Owner NANJING ANJI BIOLOGICAL TECH CO LTD
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