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Controlled-release recombinant human bone morphogenic protein-2 poly-L-lactic acid microspheres and preparation method thereof

A morphogenetic protein and lactic acid technology, which is applied to peptide/protein components, bone diseases, drug combinations, etc., can solve the problems of short release period, protein inactivation and denaturation, and difficult removal of organic solvents, and achieve the effect of mild preparation conditions

Inactive Publication Date: 2010-09-08
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are still many problems in the encapsulation of rhBMP-2 by these materials, such as the biological safety of materials, the sustained release period is too short, and it is difficult to preserve the physiological activity of rhBMP-2, so the bioavailability of local or systemic application is low
In addition, the traditional process is complicated, the cycle is long, the organic solvent is difficult to remove, the particles are large and the distribution range is wide. The main problem with the methods and processes of compounding growth factors into polymers is that these methods and processes may cause the loss of protein activity.
Usual high temperature, ultrasound, and organic solvents may cause protein inactivation and denaturation

Method used

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  • Controlled-release recombinant human bone morphogenic protein-2 poly-L-lactic acid microspheres and preparation method thereof
  • Controlled-release recombinant human bone morphogenic protein-2 poly-L-lactic acid microspheres and preparation method thereof
  • Controlled-release recombinant human bone morphogenic protein-2 poly-L-lactic acid microspheres and preparation method thereof

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Embodiment approach

[0019] As an embodiment of the present invention, a sustained-release recombinant human bone morphogenetic protein-2 poly-L-lactic acid microsphere, the microsphere matrix component is poly-L-lactic acid, and the encapsulated drug is recombinant human bone morphogenetic protein-2, poly-L-lactic acid The mixed organic solution with recombinant human bone morphogenetic protein-2 is subjected to supercritical fluid carbon dioxide extraction solvent to separate out the microsphere crystals, the particle diameter of the microspheres ranges from 825nm to 1280nm, and the encapsulation efficiency and drug loading capacity are respectively (98.7 ±1.2)% and [(24.0±0.1)×10 -3 ]%, the average drug release concentration in vitro was (3.1±0.54) μg / mL, 4.97%±1.8% during the burst release period, and the release rate reached 92.99±7.01% after 26 days, and the released recombinant human bone morphogenetic protein-2 The active protein has a complete structure and exists in the form of homodimer...

Embodiment 1

[0025] Embodiment 1 (as figure 1 shown):

[0026] 1) CO from the cylinder 2 After being liquefied by the refrigeration system, it is pressurized by a high-pressure plunger pump, heated by a constant temperature water bath in the pipeline, and pumped into a 500ml autoclave through the outer channel of the coaxial two-flow nozzle on the top of the autoclave. When the required pressure is reached in the kettle, maintain the CO 2 Pump in at a rate of 300ml / min, open the vent valve to vent at a certain rate, and keep the pressure inside the kettle constant at 12MPa. Adjust the temperature of the external drying oven and pipeline water bath of the autoclave to keep the temperature inside the autoclave constant at 33°C. When the temperature required for the experiment was 33°C, 12 mg of recombinant human BMP-2 (rhBMP-2) was dissolved in 1.2 ml of dimethyl sulfoxide, and 480 mg of poly-L-lactic acid (PLLA) was dissolved in 58.8 ml In dichloromethane (DCM), when the two solutions are...

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Abstract

The invention discloses controlled-release recombinant human bone morphogenic protein-2 poly-L-lactic acid microspheres and a preparation method thereof, which relate to a medicament carrier and preparation thereof. The component of a microsphere substrate is poly-L-lactic acid (PLLA); the wrapped medicament is recombinant human bone morphogenic protein-2 (rhBMP-2); and the released rhBMP-2 active protein is integral in structure and exists in a form of a homologous dipolymer. The preparation method comprises the following steps: mixing rhBMP-2 organic solution and PLLA organic solution to obtain mixed solution; precipitating microsphere crystals from the mixed solution after the mixed solution passes through a supercritical fluid CO2 solvent extraction; and after obtaining the microspheres, stopping pumping the mixed solution, keeping a pressure and a temperature in a high pressure autoclave unchanged, and continuously introducing CO2 to elute the microsphere crystals to remove residual organic solution. The method of the invention is simple in process, short in period, easy in organic solution removal, high in biological safety, short in burst releasing period and long in obtained microsphere particle controlled-release period, and easier in the storage of the physiological activity of the rhBMP-2.

Description

technical field [0001] The invention relates to a drug carrier and its preparation, in particular to a slow-release recombinant human bone morphogenetic protein-2 poly-L-lactic acid microsphere and its preparation method. Background technique [0002] The development and application of exogenous growth factors has always been one of the most active topics in the fields of regenerative medicine and tumor biotherapy. The half-life of active growth factors in the body is short, systemic or local application will be diluted and metabolized quickly, and direct application cannot meet the needs of tissue repair, regeneration and treatment. When using suitable carrier materials to carry growth factors into the body, it can sustainably maintain its biological activity and prolong the action time, which is attracting more and more attention and attention. With the development of genetic engineering and biotechnology, more and more polypeptide drugs are used to treat and prevent dise...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/16A61K38/18A61K47/34A61P19/08
Inventor 吴江郭应强何学令康云清尹光福陈槐卿
Owner SICHUAN UNIV
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