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A kind of intracellular pH-responsive polylactic acid nano-microspheres and preparation method thereof

A polylactic acid and nano-micro technology, applied in the field of pharmaceutical preparations of medical engineering, can solve the problems of unevenness, insufficient immune response, slow release of vaccine adjuvant antigens, etc., and achieve the effects of uniform particle size and simple preparation process.

Active Publication Date: 2020-10-02
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The present invention aims at the inhomogeneous preparation of the traditional antigen-embedded nano-microsphere vaccine adjuvant, the slow release of the antigen as a vaccine adjuvant, which is not enough to cause an effective immune response, etc., using a rapid membrane emulsification method combined with a solvent volatilization / extraction method to prepare uniform Controllable polylactic acid-based antigen-loaded nanospheres with a new structure and intracellular response release. By adjusting the internal structure of the nanospheres and co-embedding materials that help respond Enzyme releases antigen rapidly within the body, inducing higher levels of cellular and humoral immune responses

Method used

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  • A kind of intracellular pH-responsive polylactic acid nano-microspheres and preparation method thereof
  • A kind of intracellular pH-responsive polylactic acid nano-microspheres and preparation method thereof
  • A kind of intracellular pH-responsive polylactic acid nano-microspheres and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0060] Prepare 1.9% aqueous phase polyvinyl alcohol (PVA) as the outer water phase (W2), prepare 1mg / mL sodium bicarbonate aqueous solution and add a certain amount of OVA antigen (OVA final concentration is 5mg / mL) as the inner water phase (W1 ), the polylactic acid-polyglycolic acid copolymer (PLGA) that takes 0.264g is dissolved in the organic solvent methylene chloride of 8mL, as the oil phase (O), the inner aqueous phase that 4mL dissolves is added in the 8mL oil phase, Prepare water-in-oil primary emulsion (W1 / O) by homogenization or ultrasonic emulsification, then pour it into 60mL water phase containing polyvinyl alcohol (PVA), and after simple stirring, get water-in-oil-in-water type pre-emulsion (W1 / O / W2), transfer the pre-multiplex emulsion to the storage tank of the fast membrane emulsifier, select a 1.4 μm membrane, and pass the membrane 3 times under 1.5Mpa nitrogen pressure to prepare a uniform composite emulsion (W1 / O / W2), and then placed in a fume hood to st...

Embodiment 2

[0062] Prepare 1.5% aqueous poloxamer as the external aqueous phase (W2), prepare 5 mg / mL aqueous sodium bicarbonate solution and add a certain amount of HPV antigen (HPV final concentration is 10 μg / mL) as the internal aqueous phase (W1), A certain amount of polylactic acid-polyglycolic acid copolymer (PLGA) is dissolved in ethyl acetate as the oil phase (O), and 5 mL of the dissolved internal water phase is added to 10 mL of the oil phase, and homogenized or ultrasonically emulsified to prepare a water-in-oil type First emulsion (W1 / O), and then pour it into 75mL poloxamer aqueous phase, after simple stirring to obtain water-in-oil-in-water type pre-multiplex emulsion (W1 / O / W2), transfer the pre-multiplex emulsion to the fast In the storage tank of the membrane emulsifier, a 2.8μm membrane was selected and passed through the membrane three times under a nitrogen pressure of 1.2Mpa to prepare a double emulsion (W1 / O / W2) with uniform particle size, and then placed in a fume hoo...

Embodiment 3

[0064] Prepare 1.2% aqueous phase carboxymethylcellulose as the outer water phase (W2), prepare 2.5 mg / mL aqueous sodium bicarbonate solution and add a certain amount of HBsAg antigen (HBsAg final concentration is 10 mg / mL) as the inner water phase ( W1), a certain amount of polylactic acid copolymer (PLA) is dissolved in chloroform as the oil phase (O), and 6 mL of the dissolved internal water phase is added to 12 mL of the oil phase, and homogenized or ultrasonically emulsified to prepare a water-in-oil type primary Emulsion (W1 / O), and then pour it into 90mL carboxymethylcellulose water phase, after simple stirring to obtain water-in-oil-in-water type pre-complex emulsion (W1 / O / W2), transfer the pre-complex emulsion to the fast In the storage tank of the membrane emulsifier, a 30 μm membrane was selected and passed through the membrane three times under a nitrogen pressure of 1.0Mpa to prepare a double emulsion (W1 / O / W2) with uniform particle size, and then placed in a fume ...

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Abstract

The invention provides intracellular pH-response polylactic-acid nanometer microspheres and a preparing method thereof. The intracellular pH-response polylactic-acid nanometer microspheres are prepared with the rapid membrane emulsification method in cooperation with the solvent evaporation / extraction method; as the proportion between water phase materials and oil phase materials in the nanometer microspheres is adjusted, the polylactic-acid nanometer microspheres prepared with the method are of thin-skin large-cavity structures and are even in particle diameter, and have the behavior of releasing embedded substances in a responding mode in the acid environment of intracellular endosome and lysosome. The intracellular pH-response polylactic-acid nanometer microspheres can controllably and rapidly release antigens in antigen presenting cells, antigen presenting cells and T cells can be more effectively activated, the problems that the antigen releasing speed of antigen-embedding polylactic-acid nanometer microspheres prepared with the traditional method is slow, and the particle diameter of the prepared nanometer microspheres is uneven are solved, the antigen loading capacity is high, preparing is easy, and the nanometer microspheres are hopefully developed into a novel vaccine delivery and adjuvant system.

Description

technical field [0001] The invention relates to the field of pharmaceutical preparations in medical engineering, in particular to an antigen-loaded polylactic acid nano-microsphere with uniform particle size and intracellular pH-responsive release behavior and a preparation method thereof. Background technique [0002] Intracellular infection, including infectious diseases caused by viruses such as HIV and HBsAg, has always threatened human health. According to WHO data, there are still a large number of human beings infected around the world. Therefore, effective prevention and treatment of intracellular infection has important practical significance. Although the current clinically used anti-virus drugs for intracellular infection can inhibit virus replication to a certain extent, they cannot completely eliminate the infected virus in the cells, and they are prone to relapse after drug withdrawal. Long-term drug use is also likely to cause viral drug resistance. Vaccinati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61K47/34A61K9/51A61K39/12A61K39/29A61K48/00A61K39/145A61P31/20A61P31/16
Inventor 王连艳马光辉刘婍杨婷媛周炜清
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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