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Immunogenic composition

A technology of immunogenicity and composition, which is applied in the field of immunogenic composition, can solve the problems of inconsistency in performance, slow-release performance, inference of adjuvant function, and failure to realize it, and achieve a powerful effect

Active Publication Date: 2013-04-17
TORAY IND INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, regarding the mechanism by which the antigen-containing microparticles function as an adjuvant, in combination with the function of sustained release of the antigen molecule, the mechanism by which the antigen-containing microparticles together with the particles are taken up by immune cells and releases the antigen in the cells is considered to be important, And it is considered that the function of drug release from the particles is inconsistent with the performance as an adjuvant, so it is difficult to infer the adjuvant function from the sustained release performance of the particles, although it is expected to develop an effective adjuvant with performance far exceeding that of aluminum adjuvants , but this adjuvant has not been achieved so far in the technology using existing microparticles

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Example 1 Synthesis of dextran-poly(lactic acid-glycolic acid) (PLGA)

[0105] (1-1) Synthesis of TMS-dextran (compound (1))

[0106] Add dextran (Nacalai Tesque Co., Ltd., Nacalai standard super qualified product, number average molecular weight: 13,000, 5.0 g) into formamide (100 ml), and heat to 80°C. To this solution was added dropwise 1,1,1,3,3,3-hexamethyldisilazane (100 ml) over 20 minutes. After completion of the dropwise addition, the mixture was stirred at 80° C. for 2 hours. After completion of the reaction, the reaction solution was returned to room temperature, and the two layers were separated with a separatory funnel. After concentrating the upper layer under reduced pressure, methanol (300 ml) was added, and the obtained solid was filtered and dried to obtain TMS-dextran (compound (1)) (11.4 g) as a white solid.

[0107] (1-2) Synthesis of dextran-PLGA (compound (2))

[0108] Compound (1) (0.5 g) and potassium tert-butoxide (35 mg) were heated and dr...

Embodiment 2

[0109] Example 2 Preparation of Immunogenic Microparticles (Particles (1)-(10) and Comparative Particles (1), (2))

[0110] 5 mg of dextran-poly(lactic-glycolic acid) (PLGA) (compound (2)) of Example 1 was dissolved in 100 μl of dimethyl carbonate to prepare a 50 mg / ml amphiphilic polymer solution (B). After adding 20 μl of tert-butanol to the amphiphilic polymer solution (B), 50 μl of 0.025% (w / v) CEA (carcinoembryonic antigen) (COSMO BIO)) aqueous solution (A) was added dropwise, and stirred with a VORTEX mixer Make an inverse emulsion.

[0111] The inverse emulsion was pre-frozen with liquid nitrogen, and then freeze-dried for 24 hours with a freeze dryer (EYELA, FREEZEDRYERFD-1000) at a trap cooling temperature of -45°C and a vacuum of 20 Pa. The obtained solid content was dispersed in 200 µl of dimethyl carbonate to prepare a dispersion (C). Add the dispersion (C) dropwise to 2 ml of an aqueous solution containing 10% (w / v) of the surface modifier (polyvinyl alcohol or ...

Embodiment 3

[0117] Example 3 Determination of Antigen Embedding Rate in Immunogenic Microparticles

[0118]

[0119] The immunogenic microparticles (particles (3) to (7) and comparative particle (2)) produced by the method of Example 2 were suspended in 200 μl of phosphate-buffered saline to prepare a particle suspension. 100 µl of the particle suspension was transferred to an Eppendorf tube, 1 ml of distilled water was added, and the particles were precipitated by centrifugation at 13,000 rpm for 10 minutes. After removing the supernatant, the particles were resuspended in 1 ml of distilled water, and the particles were re-sedimented by centrifugation under the above conditions. After removing the supernatant, the particles were dissolved in 0.5 ml of a mixed solution obtained by mixing dichloromethane and acetone at a ratio of 1:3. The particle solution was centrifuged at 13,000 rpm for 10 minutes to precipitate CEA. After removing the supernatant, the precipitate was dissolved in 0...

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Abstract

The immunogenic composition comprises, as active ingredients, immunogenic particles obtained from antigen-adjuvant microparticle complexes wherein the antigen is enclosed in adjuvant microparticles obtained from an amphiphilic polymer having a poly(hydroxy acid) for the hydrophobic segments, and a surfactant. The immunogenic composition obtained by the surfactant being enclosed in the immunogenic microparticles has an excellent ability to activate immunity in vivo even with small amounts of antigen and few immunizations.

Description

technical field [0001] The present invention relates to an immunogenic composition. The immunogenic composition uses immunogenic microparticles as active ingredients. The immunogenic microparticles contain antigen-adjuvant microparticle complexes and surfactants. Embedded in adjuvant particles comprising an amphiphilic polymer. Background technique [0002] Adjuvants are used together with the antigen in order to increase the immune activation ability of the antigen. It is known that complete Freund's adjuvant (CFA) has a high effect as an adjuvant, but CFA formed from dead bacteria and oil emulsion has a strong side effect, and a strong inflammatory reaction and ulcerative swelling occur at the administration site (Granuloma), etc., so there are concerns about safety, and it is not allowed to be used for humans. Therefore, adjuvants that allow administration to humans are limited. [0003] As an adjuvant allowed to be administered to humans, there is an aluminum hydroxid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39
CPCA61K39/0011A61K2039/6093A61K39/39A61K47/50A61K9/16A61K9/1623A61K9/1641A61K9/1647A61K9/1652A61K9/1682A61K2039/55583A61K2039/6087
Inventor 古志洋一郎西尾玲士柿泽资训
Owner TORAY IND INC
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