Sustained-release microsphere preparation containing recombinant hepatitis B surface antigen and preparation method thereof

A technology of hepatitis B surface antigen and slow-release microsphere preparations, which is applied in the direction of medical preparations containing active ingredients, antiviral agents, and medical preparations with non-active ingredients. It can solve problems such as affecting immune effects and antigen denaturation, and achieves Effects of inducing cellular immunity, high encapsulation efficiency, and high immune activity

Active Publication Date: 2018-08-28
华北制药金坦生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to provide a slow-release microsphere preparation of recombinant hepatitis B surface antigen for a single injection, to overcome the problem that the immunogenicity of hepatitis B surface antigen is destroyed during the preparation and release of microspheres, resulting in antigen denaturation and thus affecting the immune effect. question

Method used

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  • Sustained-release microsphere preparation containing recombinant hepatitis B surface antigen and preparation method thereof
  • Sustained-release microsphere preparation containing recombinant hepatitis B surface antigen and preparation method thereof
  • Sustained-release microsphere preparation containing recombinant hepatitis B surface antigen and preparation method thereof

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Experimental program
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Embodiment 1

[0033] Trehalose and human serum albumin were selected as stability protectors, multiple groups of 0.5 mg / ml HBsAg solutions were taken, trehalose was added to make the concentrations reach 5 mg / ml and 10 mg / ml respectively; human serum albumin ( HSA) and make the concentrations reach 0.5 mg / ml, 1.0 mg / ml, 2.5 mg / ml, 5.0 mg / ml respectively, then freeze-dry in vacuum for 12 hours, and measure the HBsAg antigen activity retention rate by double antibody sandwich ELISA method, the results are shown in the table 1. In Table 1, lyophilization represents vacuum freeze-drying for 12 hours.

Embodiment 2

[0035] Trehalose and human serum albumin were selected as stability protectors, multiple 0.2 ml HBsAg solutions with a concentration of 0.5 mg / ml were taken, and trehalose and human serum albumin were respectively added to obtain trehalose concentrations of 5.0 mg / ml, human Serum albumin concentration of 1.0 mg / ml, 2.5 mg / ml, 5.0 mg / ml, 10.0 mg / ml HBsAg solution, then mixed with dichloromethane, emulsified at 15000 rpm for 150 seconds, centrifuged at 5000 rpm and taken After the supernatant was volatilized with dichloromethane, the HBsAg antigen activity retention rate was measured by double-antibody sandwich ELISA. See Table 1 for test data. The aforementioned emulsification step is represented in Table 1 by emulsification.

[0036] Table 1 Effect of stability protector on the retention rate of antigenic activity of HBsAg solution after lyophilization and emulsification

[0037]

Embodiment 3

[0039] Human serum albumin (HSA) was used as a stability protector for the preparation of HBsAg sustained-release microspheres, HBsAg and human serum albumin were mixed at a mass ratio of 1:5, and the sodium phosphate solution with a pH value of 5.0 was used as a solvent. The sodium phosphate solution was mixed as the internal aqueous phase. In this mixed solution, the concentration of HBsAg is 1 mg / ml, and the concentration of human serum albumin is 5 mg / ml. Take 100 μl of this solution and add it to 2 ml of dichloromethane and acetone containing 150 mg of 23kDa polylactic acid-polyglycolic acid block copolymer (the mass ratio of polylactic acid to polyglycolic acid (PLA:PLG) is 50:50) (65:35) mixed solution, use a high-speed homogenizer to homogenize it for 300 seconds at a speed of 20,000 rpm to form a stable W / O emulsion, and slowly inject the emulsion into the In 20ml of sodium phosphate (pH5.0) buffer solution containing NaCl and 2.5% PVA, the high-speed homogenizer was...

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Abstract

The invention discloses a slow-release microsphere preparation containing recombinant hepatitis B surface antigen and a preparation method thereof. The slow-release microsphere preparation is composed of a stability protecting agent and recombinant hepatitis B surface antigen particles, and is made of polymer material polylactic acid- The polyglycolic acid block copolymer is combined as the outer envelope layer; the stabilizer is trehalose and human serum albumin. Its preparation method is as follows: (1) mixing the stabilizer and recombinant human hepatitis B surface antigen to obtain the inner water phase; (2) dissolving the polylactic acid-polyglycolic acid block copolymer in an organic solvent to form an oil phase, and taking the inner water phase (3) Add the W / O primary emulsion to the outer water phase, stir and homogenize to form a W / O / W complex emulsion; stir for 4-6 hours, wash by centrifugation, collect, and vacuum freeze-dry That's it. The invention can overcome the problem that the immunogenicity of the hepatitis B surface antigen is destroyed during the preparation and release of the microspheres, leading to the denaturation of the antigen and thus affecting the immune effect.

Description

technical field [0001] The invention relates to a biological preparation, in particular to a slow-release microsphere preparation containing recombinant hepatitis B surface antigen and a preparation method thereof. Background technique [0002] Hepatitis B is a worldwide infectious disease that seriously threatens human health. It is a disease caused by hepatitis B virus (HBV) infection. Currently, hepatitis B vaccination is one of the most effective measures to prevent and control HBV transmission. The most commonly used hepatitis B vaccine is an aluminum adjuvanted hepatitis B vaccine made by purifying, inactivating and adding aluminum adjuvant to the hepatitis B surface antigen (HBsAg) obtained by genetic recombination technology. However, in order to achieve a lasting and effective immune response, aluminum adjuvanted hepatitis B vaccine needs to be immunized three times at 0, 1, 2 months or 0, 1, 6 months. Due to the complexity of the immunization program, a considerab...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/29A61K47/34A61K47/42A61K47/36A61K9/16A61P1/16A61P31/20
Inventor 马东杰贺进田许文娟侯丽媛王玮高珊张红霞陈会珍刘海侠张彩乔白燕张丽丽李岩异李宏进王英张卫婷
Owner 华北制药金坦生物技术股份有限公司
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