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VEGF slowly releasing injection microsphere support and its preparation and use

A microsphere, sustained-release technology, applied in the field of medicine, can solve the problems of short half-life, fast degradation, and limited clinical application.

Inactive Publication Date: 2009-02-04
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, similar to other growth factors, VEGF degrades too quickly in vivo due to its short half-life (Jeffrey L.Cleland Ph.D, Eillen T.Puenas et al.Cuthbertson Development of poly-(D,L-Lactide-co-glycolide) microspheres fermulation containing recombinant human vascular endothelial growth factor to promote local angiogenesis[J].controlled Release 2001,72:13), thus limiting its clinical application

Method used

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  • VEGF slowly releasing injection microsphere support and its preparation and use
  • VEGF slowly releasing injection microsphere support and its preparation and use
  • VEGF slowly releasing injection microsphere support and its preparation and use

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1: Preparation of VEGF slow-release microsphere scaffold by w / o / w solvent evaporation method

[0030] Dissolve 100 mg of PLGA (PLA:PGA=75:25, Mw=10,000) in 3.2 ml of dichloromethane to make an oil phase, dissolve 3 μg of VEGF and 100 mg of ammonium bicarbonate in 1 ml of redistilled water (containing 3% trehalose, 5 % mannitol) to form the inner water phase, which is added to the above-mentioned oil phase, homogenized to form w / o colostrum, 120ml of 0.1% PVA solution is placed in a stirring vessel, and the colostrum is stirred (450rpm) Quickly add it into the external water phase to fully homogenize. After five minutes, reduce the speed to 200rpm and add 30ml of distilled water to the external water phase. Stir at room temperature for 4 hours. After the microspheres are hardened, filter, wash, and freeze-dry. After sealing and dispensing, it can be irradiated and sterilized. The particle size is about 500 μm.

Embodiment 2

[0031] Embodiment 2: The w / o / w solvent volatilization method that medicine is added in the form of micropowder prepares VEGF slow-release microsphere stent

[0032] Disperse 1.2 mg of PEG (PEG6000), 300 μg of VEGF and protective agent (600 μg of zinc carbonate) in 1 ml of double-distilled water, vortex and mix for about 3 minutes, subpackage, freeze-dry, wash with dichloromethane, centrifuge, remove PEG, and obtain VEGF Micronized. Dissolve 100 mg of PLGA (PLA:PGA=75:25, Mw=10000) in 3.2 ml of dichloromethane to make an oil phase, and dissolve 100 mg of ammonium bicarbonate in 1 ml of redistilled water (containing 3% trehalose, 5% manna Alcohol) to form the inner water phase, add it to the above oil phase, homogenize for three minutes, add micronized medicine and homogenize at a low speed for one minute to form w / o colostrum, put 120ml of 0.1% PVA solution in In a stirring container, quickly add colostrum into the external water phase under stirring (450rpm) to fully homogeni...

Embodiment 3

[0033] Embodiment 3: VEGF sustained-release microspheres are tested through in vitro release

[0034] Instruments: 0508-2 desktop low-speed centrifuge (Shanghai Medical Instrument Co., Ltd.); XW-80 vortex mixer (Shanghai First Medical College Instrument Factory); CARY 100 UV spectrophotometer (Varian, USA); CS501 super constant temperature Water bath (Shanghai Pudong Rongfeng Scientific Instrument Co., Ltd.), FA1004 1 / 10,000 electronic balance (Shanghai Balance Instrument Factory);

[0035] Method and operation: Accurately weigh about 20 mg of drug-containing microspheres or blank microspheres and place them in a 7ml centrifuge tube, add 1.5ml of 10mM pH 7.2 phosphate buffer (containing 0.02% sodium azide as a bacteriostatic agent, 0.02% F -68 as a wetting agent), placed in a constant temperature water bath shaker at 37°C, with an oscillation speed of 100rpm. Take out the centrifuge tubes on day 1 and day 3 respectively, place them in a centrifuge at 2000rpm for 5min, carefully...

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Abstract

The VEGF slowly releasing injection microsphere support includes matrix, VEGF, protectant and ammonium bicarbonate. It features the matrix of polylactic acid-hydroxyl acetate copolymer of polylactic acid / hydroxyl acetate ratio 25:75 to 75:25; the protectant comprising zinc carbonate, serum albumin, mycose and mannitol; the weight ratio between VEGF and polylactic acid-hydroxyl acetate copolymer of 1:100000 to 1:10; and the microsphere size of 50-1000 microns. It is prepared through one W / O / W solvent volatilizing process or one fine medicine powder adding W / O / W solvent volatilizing process. The preparation process is simple, easy in operation and high in repeatability. In the microsphere support, protein medicine VEGF may be in vitro released for over 4 weeks in approached zero order kinetic mode. The present invention may be used in the repair and treatment of different kinds of tissue and blood vessel damage.

Description

technical field [0001] The invention relates to the technical field of medicine, and relates to a protein-loaded drug VEGF (vascular endothelial growth factor, VEGF) slow-release injection microsphere stent and a preparation method and application thereof. Background technique [0002] Vascular endothelial growth factor (vascular endothelial growth factor, VEGF) is a growth factor that specifically acts on vascular endothelial cells, can promote the proliferation, metastasis and differentiation of new blood vessels, and because VEGF can enhance vascular permeability , so it is also called vascular permeability factor. It was first purified from the conditioned medium of bovine pituitary follicular stellate cells by Ferrara et al. in 1989. Recombinant human vascular endothelial growth factor (rhVEGF) in the treatment of ischemic diseases has entered the clinical trial stage from extensive preclinical research and is expected to become a therapeutic drug for clinical applicat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/34A61K47/42A61K9/16A61K38/18A61P9/00A61P43/00
Inventor 钟延强齐琰鲁莹史国兵樊莉邓莉
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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