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Sustained-release micro-spheres preparation containing recombined erythropoietin and preparation method and use thereof

A technology for erythropoietin and sustained-release microsphere preparation, which is applied in the field of lactic acid-glycolic acid block copolymer sustained-release microsphere preparation and its preparation, and can solve the problems of immune response, pure red aplastic anemia, drug loss of efficacy, etc. , to achieve good biocompatibility, high drug loading and encapsulation efficiency, and good protein stability

Inactive Publication Date: 2009-08-19
HEBEI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Denaturation of proteins, especially pharmaceutical proteins, not only causes the drug to lose its efficacy, but also may cause immune reactions, such as the denaturation and aggregation of recombinant erythropoietin may cause pure red aplastic anemia

Method used

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  • Sustained-release micro-spheres preparation containing recombined erythropoietin and preparation method and use thereof
  • Sustained-release micro-spheres preparation containing recombined erythropoietin and preparation method and use thereof
  • Sustained-release micro-spheres preparation containing recombined erythropoietin and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Pre-freeze 10ml of a mixed solution of 0.5mg / ml recombinant human erythropoietin, 10mg / ml human serum albumin (HSA), and 45mg / ml polyethylene glycol at -80°C overnight, and routinely freeze-dry to obtain The obtained solid is continuously washed three times with dichloromethane to remove polyethylene glycol, and the mixed particles of recombinant human erythropoietin and human serum albumin are prepared. Emulsify the mixed particles of 40mg recombinant human erythropoietin and human serum albumin in 2ml of lactic acid-glycolic acid block copolymer dichloromethane solution with a concentration of 60mg / ml, under the condition that the stirring rate is 20000 rpm Emulsified for 60s to obtain S / O emulsion. Inject the emulsion into 250ml of 0.02M phosphate buffer (pH7.4) containing 2% polyvinyl alcohol with a micro sampler, stir at a speed of 600rpm for 1min to prepare the S / O / W emulsion, and add the emulsion to the emulsion Add 250ml 0.02M phosphate buffer solution to the s...

Embodiment 2

[0041] Pre-freeze 10ml of the mixed solution of 0.5mg / ml recombinant erythropoietin, 10mg / ml human serum albumin (HSA), and 30mg / ml polyethylene glycol at -80°C overnight, and routinely freeze-dry.

[0042] The obtained solid was washed continuously with dichloromethane for three times to remove polyethylene glycol, and mixed particles of recombinant erythropoietin and human serum albumin were prepared. The mixed particles of 40mg recombinant erythropoietin and human serum albumin were emulsified in 2ml of 120mg / ml lactic acid-glycolic acid block copolymer dichloromethane solution, and emulsified at a stirring rate of 20000 rpm 60s, get S / O emulsion. Inject the emulsion into 250ml of 0.02M phosphate buffer (pH is 7.4) containing 2% polyvinyl alcohol with a micro sampler, and stir at a speed of 600rpm for 1min to prepare the S / O / W emulsion. Add 250ml of 0.02M phosphate buffer solution to the solution, stir at 300rpm at room temperature for 6h, volatilize the organic solvent, c...

Embodiment 3

[0044] Pre-freeze 10ml of a mixed solution of 0.5mg / ml recombinant erythropoietin, 10mg / ml human serum albumin (HSA), and 45mg / ml polyethylene glycol at -80°C overnight, and routinely freeze-dry to obtain The solid matter was continuously washed with dichloromethane three times to remove polyethylene glycol, and the mixed microparticles of recombinant erythropoietin and human serum albumin were prepared. 40 mg of recombinant human erythropoietin and human serum albumin mixed particles were emulsified in 2 ml of 150 mg / ml lactic acid-glycolic acid block copolymer dichloromethane solution at a stirring rate of 20,000 rpm Emulsified for 60s to obtain S / O emulsion. Inject the emulsion into 250ml of 0.02M phosphate buffer (pH7.4) containing 2% PVA with a micro sampler, stir at a speed of 400rpm for 1min to prepare the S / O / W emulsion, and add Add 250ml of 0.02M phosphate buffer, stir at 300rpm at room temperature for 6h, volatilize the organic solvent, collect the microspheres by c...

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Abstract

The invention discloses a slow-release microsphere preparation containing a recombined human haemopoietin. The slow-release microsphere preparation is prepared by the S / O / W compound emulsion solvent volatilization method. The preparation method comprises that: firstly, by the freeze-drying method, a micro particle containing the human serum albumin and the recombined human haemopoietin is prepared; secondly, by using a biodegradable high molecular material of lactic acid-glycollic acid block copolymer as a carrier material, the micro particle containing the human serum albumin and the recombined human haemopoietin is encapsulated; and thirdly, the lactic acid-glycollic acid block copolymer slow-release microsphere preparation containing the recombined human haemopoietin is prepared. The microsphere of the invention has the advantages of smooth surface, uniform appearance, regular size and no adhesion, the average particle size is between 70 and 105mu m; moreover, the microsphere is high in drug-carrying quantity and encapsulating rate, and the in vitro slow release period is more than 30 days. The obtained slow-release microsphere preparation is good in biocompatibility and can be used for non-intravenous drug administration such as hypodermic drug administration and intramuscular drug administration, and when used as a drug preparation for treating renal anemia, the slow-release microsphere preparation can improve the hematocrit of a patient.

Description

technical field [0001] The invention relates to a biological preparation, in particular to a lactic acid-glycolic acid block copolymer slow-release microsphere preparation containing recombinant human erythropoietin and a preparation method thereof. Background technique [0002] Erythropoietin (EPO) is an active glycoprotein synthesized by the renal proximal tubule, which can stimulate the proliferation and differentiation of bone marrow erythroid hematopoietic blasts, and play a major role in regulating the development of red blood cells. At present, recombinant human erythropoietin produced by genetic engineering technology has become the first choice drug for clinical treatment of renal anemia, and has good effects on anemia caused by tumor chemotherapy and other anemia diseases. However, the currently clinically used dosage form is powder injection or water injection of erythropoietin for intramuscular or intravenous injection. Current treatment for all approved indicat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/16A61K38/18A61K47/48A61K47/42A61K47/34A61P7/06A61K47/60
Inventor 贺进田冯美彦姜杨梁倩
Owner HEBEI NORMAL UNIV
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