Multifunctional fused polypeptide as well as preparation method and application thereof

A fusion polypeptide and multifunctional technology, which is applied in the field of medicine, can solve the problems of poor blood vessel specificity and selectivity, and unclear targets of multifunctional fusion polypeptides, so as to prevent or treat rheumatoid arthritis, prevent Or treat retinal neovascular eye disease and improve the effect of neovascularization

Active Publication Date: 2014-09-17
NANJING ANJI BIOLOGICAL TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] Aiming at the problems of the existing multifunctional fusion polypeptides with unclear targets and poor specificity and selectivity for blood vessels, the present invention provides a multifunctional fusion polypeptide and its preparation method and application, which can treat various entities Tumors, rheumatoid arthritis and neovascular eye diseases have good preventive and therapeutic effects, increasing the scope of its indications

Method used

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  • Multifunctional fused polypeptide as well as preparation method and application thereof
  • Multifunctional fused polypeptide as well as preparation method and application thereof
  • Multifunctional fused polypeptide as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1 Preparation and inspection of multifunctional fusion polypeptide polypeptide

[0058] The polypeptide was synthesized by solid-phase synthesis, separated and purified by preparative HPLC, and the purity of the polypeptide was determined by analytical RP-HPLC.

[0059] Polypeptide I-VI solid-phase synthesis method uses Fmoc-wang-resin or Fmoc-CTC-resin (Gill Biochemical Co., Ltd.) as the starting material, and then uses protected amino acids to connect dipeptides to tetrapeptides in sequence, and the peptide connection work is completed After that, fully wash, then cut the peptide, and post-process the crude product of the multifunctional fusion polypeptide. The crude product was dissolved, purified twice by preparative high-performance liquid phase, concentrated and freeze-dried to obtain the pure product, and finally purified for the third time to obtain the refined peptide product. The method can not only ensure the synthesis efficiency, but also improve...

Embodiment 2

[0092] Inhibitory effect of multifunctional fusion polypeptide on the proliferation of human retinal vascular endothelial cells (HRCEC)

[0093] MTT assay was used to detect the activity of the multifunctional fusion polypeptide in inhibiting 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. After the cells are completely adhered to the wall, the multifunctional fusion polypeptide polypeptide is used as the administration group, Avastin is used as the positive control group, and the culture solution without any drug is used as the blank control...

Embodiment 3

[0117] Effect of angiogenesis blocking agent polypeptide on corneal neovascularization in BALB / c mice

[0118] (1) Preparation of corneal neovascularization model induced by alkali burn in BALB / c mice:

[0119] Thirty-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 NaOH solution to make it saturated, excess liquid was removed, and the filter paper sheet was placed in BALB / c 40S in the center of the mouse cornea, 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 rota...

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Abstract

The invention discloses a multifunctional fused polypeptide as well as a preparation method and application thereof, belonging to the field of bioengineering pharmaceuticals. The multifunctional fused polypeptide is a combination of one or any multiple of six polypeptides and can be used for preventing and treating solid tumors, rheumatoid arthritis and neovascular eye disease. The multifunctional fused polypeptide is prepared by adopting an artificial synthesis method and is characterized in that angiogenesis is restrained by restraining vascular endothelial cell proliferation and migration, and subsequently the angiogenesis-related diseases, such as solid tumors, rheumatoid arthritis and neovascular eye disease are treated.

Description

technical field [0001] The invention relates to the field of medicine, in particular to a class of multifunctional fusion polypeptide and its preparation method and its application in the preparation of medicines for treating solid tumors, rheumatoid arthritis and neovascular eye diseases. Background technique [0002] Angiogenesis refers to the process of forming new blood vessels by sprouting on the basis of primitive vascular plexus or existing blood vessels. Angiogenesis plays an important role in wound healing, placenta formation and embryonic development. At the same time, however, angiogenesis also occurs in a variety of pathological conditions, including: tumor growth and metastasis; inflammatory disorders such as rheumatoid arthritis, psoriasis, osteoarthritis, inflammatory bowel disease Crohn's disease, ulcerative colon, and other inflammatory disorders; ocular neovascularization, such as retinopathy of prematurity, ischemic retinopathy, retinal vein or artery occl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C07K1/06A61K38/16A61P35/00A61P29/00A61P19/02A61P27/02
Inventor 康梦实李梦玮
Owner NANJING ANJI BIOLOGICAL TECH CO LTD
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