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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: 2012-01-25
TORAY IND INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Furthermore, regarding the mechanism by which the microparticles containing the antigen function as an adjuvant, it is considered important to combine the function of the sustained release of the antigen molecule, and the mechanism by which the microparticles containing the antigen together with the particles are taken up by the immune cells to release the antigen in the cells, 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 such adjuvants have not been achieved so far using existing microparticle technology

Method used

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Examples

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Embodiment 1

[0097] The synthesis of embodiment 1 dextran-poly(lactic acid-glycolic acid) (PLGA)

[0098] (1-1) Synthesis of TMS-dextran

[0099] 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 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.

[0100] In the same manner, compounds (2) and (3) were prepared using dextran (manufactured by Sigma, average molecular weight 1,500 or less), and compounds (4) a...

Embodiment 2

[0108] The synthesis of embodiment 2PEG-PLGA (compound (10), (11))

[0109]Polyethylene glycol monomethyl ether (SUNBRIGHTMEH-20H manufactured by NOF Co., Ltd., number average molecular weight 5,128, Mw / Mn=1.02), (DL)-lactide and glycolide were mixed in the amount shown in Table 2, Heat to 140°C. After stirring for 20 minutes, tin (II) octoate (0.05% by weight based on polyethylene glycol monomethyl ether) was added, and stirred at 180° C. for 3 hours. After the reaction solution was returned to room temperature, it was dissolved in chloroform (to form a concentration of about 100 mg / ml), and purified by reprecipitation with diethyl ether cooled to 0°C, and the obtained solid was filtered off and dried under reduced pressure to obtain a white or pale The tan solid yielded PEG-PLGA polymer. The number average molecular weight of this polymer is given by 1 H-NMR determined (Table 2).

[0110] 【Table 2】

[0111] Table 2: Evaluation results of the prepared PEG-PLGA polymers ...

Embodiment 3

[0113] Example 3 Preparation of Antigen-Adjuvant Microparticle Complex Using Dex-g-PLGA Polymer (Dex-g-PLGA Particles (1)-(28))

[0114] 5 mg of dextran-poly(lactic-glycolic acid) (PLGA) (compounds (12) to (23)) of Example 1 was dissolved in 100 μl of dimethyl carbonate to prepare a 50 mg / ml polymer solution. After adding 20 μl of tert-butanol to the polymer solution, 50 μl of the embedded antigen (OVA (ovalbumin) (Sigma) or CEA (carcinoembryonic antigen) (COSMO) was added dropwise at the concentration recorded in Table 3. BIO company)) and immunoactive substances (CpG) were stirred with a VORTEX mixer to prepare an inverse emulsion. As CpG, 5'ggggggggCGACGATCGTCAgG-3' (small letters in the sequence indicate phosphorothioate modified bases) (commissioned by Sigma- Synthesized by Genosys Corporation).

[0115] The inverse emulsion was pre-frozen with liquid nitrogen, and then freeze-dried for 24 hours with a freeze dryer (EYELA, FREEZEDRYER FD-1000) at a trap cooling temperatu...

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Abstract

Disclosed is an antigen-adjuvant microparticle complex which comprises adjuvant microparticles and an antigen encapsulated in the adjuvant microparticles, wherein each of the adjuvant microparticles comprises an amphipathic polymer which has a poly(hydroxy acid) as a hydrophobic segment. Also disclosed is an immunogenic composition which contains particles conjugated with the complex as an active ingredient. The complex or the immunogenic composition can induce a high immune response to a small quantity of an antigen or can induce a high immune response to an antigen by a few frequencies of administration, and is therefore useful as a vaccine that is effective for the treatment and prevention of infectious diseases, cancer and others.

Description

technical field [0001] The present invention relates to an immunogenic composition comprising an antigen-adjuvant microparticle complex, the antigen embedded in the adjuvant microparticles comprising an amphiphilic polymer, as an active ingredient. Background technique [0002] In order to increase the immune activation ability of an antigen, an adjuvant is used together with the antigen. As an adjuvant, the adjuvant known to be highly effective is Freund's complete adjuvant (CFA), but CFA formed from dead bacteria and oil emulsion has a strong side effect, and a strong inflammatory reaction and ulceration occur at the administration site Sexual swelling (granuloma), etc., so there are concerns about safety, and it is not allowed to be used for humans. Adjuvants permitted for human administration are limited. An adjuvant allowed to be administered to humans is an aluminum hydroxide adjuvant, but it is difficult to say that the immune activation ability of this adjuvant is ...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61K47/34A61K47/36A61K47/48
CPCA61K47/48215A61K2039/6025A61K2039/55555A61K2039/545A61K47/4823A61K39/39A61K47/60A61K47/61A61P37/04
Inventor 西尾玲士井田伸夫
Owner TORAY IND INC
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