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Polypeptide for preventing and treating angiogenesis and application thereof

An angiogenesis and purpose technology, applied in the field of biomedicine, can solve the problems of cumbersome purification process, endotoxin residue, complex spatial conformation, etc.

Inactive Publication Date: 2011-01-12
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Third, based on the above main reasons, the bioavailability of ophthalmic drugs is very low; to improve it, the concentration of administration can be increased
Compounds used to treat neovascularization of tumors have obvious side effects, and cannot be administered in high doses both systemically and locally
[0009] Fourth, although a series of relatively safe endogenous angiogenesis inhibitors have been confirmed successively, such as angiostatin (angiostatin), which consists of plasminogen Kringle domains 1-4 (plasminogenKringle 1-4), Can significantly inhibit the growth of blood vessel-dependent tumors, but due to its large molecular weight and complex spatial conformation, there are deficiencies in the preparation process such as cumbersome recombinant expression and purification processes and endotoxin residues

Method used

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  • Polypeptide for preventing and treating angiogenesis and application thereof
  • Polypeptide for preventing and treating angiogenesis and application thereof
  • Polypeptide for preventing and treating angiogenesis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0122] Synthesis of Peptides

[0123] Three QY polypeptides whose sequences are shown in SEQ ID NO: 1-3 were synthesized using a commercially available SYMPHONY polypeptide synthesizer.

[0124] QY-1: CQHWSAQTPHTHNRTPENFPCKNLDENYC (SEQ ID NO: 2); the cysteine ​​residues at both ends form a disulfide bond from the beginning to the end, and the small peptide is in a circular conformation.

[0125] QY-2: CQHWSAQTPHTHNRTPENFPCKNLDENYC (SEQ ID NO: 3); in a linear conformation.

[0126] QY-3: QHWSAQTPHTHNRTPENFPCKNLDENY (SEQ ID NO: 1); in a linear conformation.

[0127] Proceed as follows:

[0128] 1. Prepare the required protected amino acid solution, condensation reagent, and cutting reagent according to the software calculation, and add enough DMF and DCM to the corresponding bottle of the instrument

[0129] 2. Add 100μmol FMOC-Ala-Wang-Resin to the reactor,

[0130] 3. Put a centrifuge tube on the pipe that collects the cutting fluid.

[0131]4. Edit the program. Generally...

Embodiment 2

[0136] Effects of QY on Angiogenesis of Chicken Embryo Allantoic Membrane

[0137] 1. Materials and methods:

[0138] 1.1 Materials: Whatman filter paper was purchased from SIGMA, USA.

[0139] 1.2 Model making and intervention test: Breeding eggs 1-2 days after birth were randomly divided into 3 groups, followed by the control group and the 50 μg QY intervention group (respectively QY1, QY2 and QY3 polypeptides), with 9-13 eggs in each group. Wash the surface of the eggs with tap water, soak them in 0.2% bromogeramine solution for 5-10 minutes, with the blunt end facing up, and cultivate them in an incubator with a temperature of 37°C and a humidity of 60%-70%, and turn them over at least twice a day to 24h is one day, hatching for 5 days. Under sterile conditions, use tweezers to open a hole of about 1cm at its blunt end (air cell end). 2 -2cm 2 Open the eggshell membrane and the air chamber in turn, revealing the allantoic membrane with blood vessels. Whatman filter pa...

Embodiment 3

[0145] Inhibitory effect of QY on VEGF-induced proliferation of RF / 6A cell line

[0146] 1. Materials and Methods

[0147] Monkey choroid-retinal (endothelial) cell line RF / 6A was purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

[0148] Cell proliferation test: Cell proliferation was detected using CellTiter 96 Aqueous One Solution Cell Proliferation Assay (MTS) kit (Promega, Madison, USA), and the operation was performed according to the manufacturer's instructions. Specifically, after the overgrown cells were digested with trypsin, they were resuspended in a 96-well plate at a density of 100,000 / mL. When the cells grew to 90% of the bottom of the well, they were replaced with fresh low-serum medium and added Different concentrations of small peptides. After 30 minutes, VEGF was added at a final concentration of 10 ng / mL. After 24 hours of incubation, 20uL of MTS was added to each well, and fresh medium w...

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Abstract

The invention relates to polypeptide for preventing and treating angiogenesis and application thereof. The invention also relates to the preparation and the application of the polypeptide and a medicinal composition containing the polypeptide. The polypeptide has a plurality of types of advantages, such as small molecular weight for permeating various eye tissue barriers, high water solubility for keeping high concentration in neutral tear, aqueous humor and vitreous humor, and the like.

Description

technical field [0001] The present invention relates to the field of biomedicine, and more specifically, relates to a novel small molecule polypeptide for preventing and treating angiogenesis, called QY. The present invention also relates to the preparation method and application of the polypeptide and the pharmaceutical composition containing the polypeptide. Background technique [0002] Angiogenesis is the process of growing new capillaries in the existing vascular system, which is a physiological change necessary for embryonic development, wound healing, tissue regeneration and repair, but it is also the pathological basis of various systemic and local diseases, including neonatal Vascular eye diseases, neovascularization of tumors, rheumatoid arthritis, psoriasis, etc. [0003] Angiogenesis involves a complex multi-step process, including proliferation, migration, erosion and lumen formation of vascular endothelial cells. Angiogenesis is tightly regulated by a balance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00C12N15/11A61K38/16A61P35/00A61P9/10A61P9/00A61P7/02A61P29/00A61P1/00A61P1/04A61P17/02A61P19/04A61P17/00A61P17/06A61P15/08A61P27/02
Inventor 许迅赵卉
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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