Integrin blocking agent polypeptide and applications of the same in preparation of drugs for treatment of neovascular eye diseases

An integrin blocker and neovascularization technology, which is applied in cardiovascular system diseases, drug combinations, peptides, etc., can solve the problems of vascular specificity and selectivity, large dosage, and unclear targets. To achieve the effect of preventing or treating neovascular eye disease

Active Publication Date: 2015-02-04
NANJING ANJI BIOLOGICAL TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] Aiming at the unclear targets of the existing angiogenesis inhibitors, their specificity and selectivity to blood vessels are not good enough, and the effect is limited, resulting in a large amount of drug used in the experiment. The present invention provides an integrin blocking Agent polypeptide and its application in the preparation of medicines for the treatment of neovascular eye diseases, which have the activity of efficiently inhibiting neovascularization, especially have a good effect on abnormal ocular vascular diseases

Method used

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  • Integrin blocking agent polypeptide and applications of the same in preparation of drugs for treatment of neovascular eye diseases
  • Integrin blocking agent polypeptide and applications of the same in preparation of drugs for treatment of neovascular eye diseases
  • Integrin blocking agent polypeptide and applications of the same in preparation of drugs for treatment of neovascular eye diseases

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Experimental program
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Effect test

Embodiment 1

[0040] Inhibitory Effect of Integrin Blocker Peptides on Human Retinal Vascular Endothelial Cells (HRCEC) Proliferation.

[0041] MTT assay was used to detect the inhibitory activity of integrin blocker polypeptide on the proliferation of human retinal vascular endothelial cells. HRCEC cells were incubated at 37°C, 5% CO 2 When cultured in an incubator with a density of more than 90%, it was digested with trypsin and collected, and the cells were resuspended in culture medium and counted under a microscope, and the cell concentration was adjusted to 3.0×10 4 cells / mL, seed the cell suspension into a 96-well plate, 100 μL per well, and inoculate at 37°C, 5% CO 2 Incubate overnight in the incubator. Dilute polypeptide I, polypeptide II, and Avastin with culture medium to each predetermined concentration. After the cells were completely attached to the wall, each dilution was added to a 96-well plate, 100 μL per well. The addition of integrin blocker polypeptide was used as t...

Embodiment 2

[0056] In vivo anti-angiogenic activity of peptides analyzed in chicken chorioallantoic membrane (CAM)

[0057]In this study, CAM assay was used to investigate the activity of integrin blocker polypeptides in inhibiting angiogenesis in vivo. Studies have shown that the biosynthesis rate of collagen reaches the maximum from the 8th day to the 11th day of chicken embryo development, which is the most vigorous stage of angiogenesis, and the body's immune system has not yet been fully established, so it is selected to develop to the 8th day The chick embryos began to be administered. Considering that the polypeptide on the drug-loaded paper sheet has a certain diffusion range limitation on the chicken embryo allantoic membrane, only the new blood vessels within a radius of 5 mm from the edge of the paper sheet and the filter membrane within an angle of less than 45 degrees were counted in the test. quantity. Take the following steps:

[0058] (1) Cultivate the Bailaihang chicke...

Embodiment 3

[0066] Effect of Integrin Blocker Peptides on Corneal Neovascularization in Mice.

[0067] (1) Preparation of corneal neovascularization model induced by alkali burn in BALB / c mice:

[0068] Twenty healthy BALB / c mice, male, weighing 20-25 g, were examined under a slit-lamp microscope for the anterior segment 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, clamp a single-layer filter paper sheet with a diameter of 2mm with tweezers, soak it in 1mol / L sodium hydroxide solution to make it saturated, remove excess liquid, and spread the filter paper sheet evenly Stick it on the central cornea of ​​BALB / c mice for 40 seconds, discard the filter paper, and immediately rinse the burn area and conjunctival sac with 15 mL of PBS for 1 min. Wipe off excess water with a cotton swab, and rotate the cor...

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Abstract

The present invention discloses an integrin blocking agent polypeptide and applications of the same in preparation of drugs for treatment of neovascular eye diseases, and belongs to the field of medicine. The integrin blocking agent polypeptide comprises a polypeptide I Arg-Gly-Asp-Gly-Gly-Gly-Gly-Phe- Gln-Pro-Val-Leu-His-Leu-Val-Ala-Leu-Asn-Ser-Pro-Leu-Ser-Gly-Gly-Met-Arg-Gly-Ile-Arg-Gly-Ala-Asp-Phe-Gln-Cys-Phe-Gln-Gln-Ala-Arg-Ala-Val-Gly-Leu-Ala-Gly-Thr-Phe-Arg-Ala and a polypeptide II Phe-Gln-Pro-Val-Leu-His-Leu-Val-Ala-Leu-Asn-Ser-Pro-Leu-Ser-Gly-Gly-Met-Arg-Gly-Ile-Arg-Gly-Ala-Asp-Phe-Gln-Cys-Phe-Gln-Gln-Ala-Arg-Al a-Val-Gly-Leu-Ala-Gly-Thr-Phe-Arg-Ala-Gly-Gly-Gly-Gly-Arg-Gly-Asp, wherein the polypeptide I is modified with the integrin ligand sequence (Arg-Gly-Asp-Gly-Gly-Gly-Gly). The integrin blocking agent polypeptide can be used for treatments of iridal neovascular eye diseases, choroidal neovascular eye diseases, retinal neovascular eye diseases or corneal neovascular eye diseases and other neovascular eye diseases.

Description

technical field [0001] The invention belongs to the field of medicines, and more specifically relates to an integrin blocker polypeptide and its application in the preparation of medicines for treating neovascular eye diseases. The integrin blocker polypeptide includes two kinds of polypeptides. Background technique [0002] Neovascularization, highly regulated under normal physiological conditions, is an essential process in reproduction, embryonic development, tissue repair and wound healing. [0003] Angiogenesis occurs in response to a wide variety of pro-angiogenic stimuli, such as growth factors, cytokines and other physiological molecules, as well as other factors such as hypoxia and low pH. The angiogenic cascade of neovascular development requires the cooperation of multiple molecules that regulate essential cellular processes such as extracellular matrix (ECM) remodeling, invasion, migration, proliferation, differentiation, and tubule formation, among others. Prov...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00A61K38/16A61P27/02A61P9/10A61P3/10
CPCA61K38/00C07K14/00Y02A50/30
Inventor 康梦实
Owner NANJING ANJI BIOLOGICAL TECH CO LTD
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