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A nanosuspension for oral protein immunization and its preparation method

A nano-suspension and protein technology, applied in the field of medicine, can solve the problems of increased vaccine intake, poor curative effect, and easy inactivation, and achieve the effects of prolonging residence time, increasing intake, and improving stability

Inactive Publication Date: 2019-01-11
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Aiming at the problems that the protein vaccine is easily inactivated after oral administration, the curative effect is poor, and the intramuscular injection of the vaccine cannot induce mucosal immune response, the present invention designs a nanosuspension for oral protein immunity, and utilizes the transfer effect of FAE on nanoparticles , transporting nanoparticles loaded with protein antigens to the vicinity of APCs such as macrophages and dendritic cells, with the help of receptor-mediated active targeting of APCs, increasing the uptake of vaccines by APCs, thereby inducing an immune response

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Preparation of PLGA / BSA nanosuspension:

[0034] (1) Accurately weigh 40mg of PLGA, dissolve it in 1mL of dichloromethane, and use it as the oil phase.

[0035] (2) Accurately weigh 40 mg of BSA, dissolve it in 1 mL of deionized water, and use it as the inner water phase.

[0036] (3) Accurately weigh 1.25 g of poloxamer 188 and dissolve in 100 mL of deionized water to obtain a 1.25% poloxamer 188 solution.

[0037] (4) Accurately weigh 0.1 g of poloxamer 188, dissolve it in 100 mL of deionized water, and use it as a diluent.

[0038] (5) The pH of 5 mL of 0.5% acetic acid solution was adjusted to about 5.5 with sodium hydroxide solution, and the volume was adjusted to 10 mL with deionized water to obtain acetate buffer.

[0039] (6) Mix 1 mL of acetate buffer solution with 4 mL of 1.25% poloxamer 188 solution to obtain an external aqueous phase.

[0040] (7) Add 0.25 mL of the inner water phase to the oil phase, and in an ice bath, the probe is ultrasonicated for 9 ...

Embodiment 2

[0044] Preparation of CS / PLGA / BSA nanosuspension:

[0045] (1) Accurately weigh 40mg of PLGA, dissolve it in 1mL of dichloromethane, and use it as the oil phase.

[0046] (2) Accurately weigh 40 mg of BSA, dissolve it in 1 mL of deionized water, and use it as the inner water phase.

[0047] (3) Accurately weigh 1.25 g of poloxamer 188 and dissolve in 100 mL of deionized water to obtain a 1.25% poloxamer 188 solution.

[0048] (4) Accurately weigh 0.1 g of poloxamer 188, dissolve it in 100 mL of deionized water, and use it as a diluent.

[0049] (5) Accurately weigh 5 mg of chitosan, dissolve it in 5 mL of 0.5% acetic acid solution, and after it is completely dissolved, adjust its pH to about 5.5 with sodium hydroxide solution, add deionized water to make it to 10 mL, and obtain the concentration 0.5mg / mL chitosan solution.

[0050] (6) Mix 1 mL of chitosan solution with 4 mL of 1.25% poloxamer 188 solution to obtain an external aqueous phase.

[0051] (7) Add 0.25 mL of th...

Embodiment 3

[0055] Preparation of MCS / PLGA / BSA nanosuspension:

[0056] This example is basically the same as Example 2, except that MCS is used instead of CS, and the concentration of MCS solution is 2 mg / mL. The obtained MCS / PLGA / BSA nanosuspension has a particle size of 532.8nm, a polydispersity coefficient of 0.184, a zeta potential of 28.92mv, and an encapsulation efficiency of 89.34%.

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PUM

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Abstract

The invention relates to a nano suspension for oral protein immunization and a preparation method thereof, and belongs to the field of medicine. Bovine serum albumin (BSA) is taken as the model drug, PLGA is taken as the carrier, and a solvent volatilization method is adopted to prepare a BSA loaded MCS / PLGA / BSA nano suspension. The particle size of the suspension is 532.8 nm, the zeta potential is 28.92 mv, and the encapsulation rate is 89.34%. Mannosylated chitosan (MCS) is adsorbed on the surface of PLGA / BSA nano particles through a static effect, thus the stability of nano particles in gastrointestinal liquid becomes more stable, moreover, the stay time in intestinal cavity is prolonged, and the nano particles with positive charges can be absorbed by cells more easily. Nano particles are transferred by epithelium related with follicle to PP nodes; through the combination between mannose ligands and mannose acceptors on antigen-presenting cells (APCs), the intake of APCs is increased, and thus the immunization reactions are induced.

Description

technical field [0001] The invention relates to a nano-suspension for oral protein immunization and a preparation method thereof, belonging to the field of medicine. Background technique [0002] Vaccines are one of the most effective ways to prevent infectious diseases. The mucosa is exposed to a large number of commensal and pathogenic microorganisms, and is the primary area for infection, and in a healthy adult, the mucosa-associated lymphoid tissue (MALT) accounts for 80% of all immune cells and is the largest mammalian lymphatic system. . However, the vast majority of marketed vaccines are intramuscular injection vaccines. Although intramuscular immunization can produce a strong systemic immune response, it cannot induce a strong immune response at the mucosal site. Therefore, compared with injectable vaccines, mucosal vaccines have many advantages, including inducing effective mucosal and systemic immune responses, producing mucosal IgA and serum IgG antibodies, avoi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/113A61K9/10A61K47/34A61K47/61A61K39/13A61K39/145
CPCA61K9/10A61K9/113A61K39/12A61K47/34A61K2039/542A61K2039/6087A61K2039/6093A61K2039/62C12N2750/14034C12N2760/16134
Inventor 王柏迟环宗莉
Owner CHINA PHARM UNIV
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