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Oil-in-water type nanometer emulsion adjuvant and MRSA nanometer emulsion adjuvant vaccine and preparing method thereof

An oil-in-water type and nanoemulsion technology, which is applied in the field of biomedicine, can solve the problems of inconspicuous enhancement of vaccine immunogenicity, long protection period, and small side effects, and achieve enhanced bioavailability, low body irritation, and The effect of small side effects

Active Publication Date: 2016-01-20
ARMY MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to overcome the existing adverse reactions of existing vaccine adjuvants, especially aluminum adjuvants, MF59, AS series and other adjuvants have no obvious enhancement effect on vaccine immunogenicity and poor stability. The present invention provides an oil-in-water adjuvant The nanoemulsion adjuvant also provides a vaccine containing the nanoemulsion adjuvant and a preparation method, which can greatly enhance the immune effect of the vaccine and reduce adverse reactions. At the same time, the composition is simple, the dosage form is stable, and it is easy to use; the vaccine prepared by the adjuvant and the antigen, Good stability, small side effects, good immune effect, long protection period

Method used

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  • Oil-in-water type nanometer emulsion adjuvant and MRSA nanometer emulsion adjuvant vaccine and preparing method thereof
  • Oil-in-water type nanometer emulsion adjuvant and MRSA nanometer emulsion adjuvant vaccine and preparing method thereof
  • Oil-in-water type nanometer emulsion adjuvant and MRSA nanometer emulsion adjuvant vaccine and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Screening of nanoemulsion adjuvant prescription

[0038] For surfactants (SF), such as: Tween-80, Tween-85, Tween 60, Tween 20, polyoxyethylene castor oil (EL40, EL35), polyoxyethylene hydrogenated castor oil, etc., help surface activity (CoSF), such as Span 80, Span 85 and ethanol, glycerin, propylene glycol, absolute ethanol, 1,3-butanol, 1,3-propanediol, comprehensively design the experimental group, according to 9:1, 8:2 , 7:3, 6:4, 5:5, 4:6, 3:7, 2:8, 1:9 (the mass ratio of SF and CoSF), weigh SF and CoSF, add the selected oil phase, Such as: paraffin oil, ethyl acetate, isopropyl myristate, squalane, squalene, ethyl oleate, caprylic acid / decanoic acid triglyceride, etc., vortex and shake, add water dropwise, observe the changes in each group, Centrifugation at 13,000 rpm for 30 minutes, stability and emulsion droplet size were the main evaluation criteria to determine the SF and CoSF that form the nanoemulsion. Use surfactant / cosurfactant (mixed surfacta...

Embodiment 2

[0060] Example 2. Preparation of MRSA nanoemulsion adjuvant vaccine with oil-in-water nanoemulsion adjuvant

[0061] Due to the wide variety of vaccines, this example mainly uses the MRSA nanoemulsion adjuvant vaccine to prove the effect of the nanoemulsion adjuvant vaccine prepared by the present invention.

[0062] The preparation of MRSA protein antigens was carried out by reference (ZuoQF, YangLY, FengQ, LuDS, DongYD, CaiCZ, et al. Evaluation of the protective immunity ofanovel subunit fusionvaccineinamurine model of systemic MRSA infection. PloSone 2013; 8: e81212). The original concentration of the protein antigen is 1.5 mg / mL, the purity of the protein is 99.4%, and the prepared antigen is stored in a refrigerator at -70°C.

[0063] According to the oil-in-water nanoemulsion adjuvant screened in Example 1, a vaccine of the oil-in-water nanoemulsion adjuvant is prepared. The specific formula and preparation method are as follows:

[0064] Formula 1: 150μg / ml MRSA nanoemulsion ad...

Embodiment 3

[0086] Example 3 Evaluation of the basic quality characteristics of MRSA nanoemulsion adjuvant vaccine

[0087] A small amount of MRSA nanoemulsion adjuvant vaccine is appropriate, diluted 100 times with water, and dropped on a copper net covered with a supporting film. After standing for 10 minutes, it is blotted dry with a filter paper, and then 2% phosphotungstic acid (pH 7.4) ) The solution was negatively dyed on a copper mesh for 3 minutes, and then evaporated to dryness. Observed by a transmission electron microscope and photographed. A small amount of MRSA nanoemulsion adjuvant vaccine is appropriate, diluted 100 times with water, then air-dried naturally, and directly observed with the atomic mechanics microscope IPC-208B (Chongqing University). All images are automatically smoothed to eliminate low-frequency noise in the slow-scan direction. The measurement is done under the following conditions: tungsten probe (force constant, 0.06N·m); scanning range, 10.5×10.5nm; ima...

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Abstract

The invention discloses an oil-in-water type nanometer emulsion adjuvant and MRSA nanometer emulsion adjuvant vaccine and a preparing method thereof. The nanometer emulsion adjuvant is prepared from, by mass, 1-30% of surface active agent, 0.1-15% of cosurfactant, 0.1-15% of oil phase and 40-98.8% of water; the adjuvant is clear and transparent liquid from appearance, the particle size ranges from 1 nm to 100 nm, viscosity is low, the adjuvant belongs to a highly thermodynamics stabilizing system, and therefore high-speed centrifugation is stable without layering; when being compatible with the vaccine, the adjuvant can wrap the vaccine or be directly mixed with the vaccine physically for use, and immunity can be achieved by means of administration routes such as intramuscular injection and nasal drip. The adjuvant has the advantages of being low in cost, convenient to administrate, small in toxicity, good in liquidity and small in irritation to organism, and avoiding cross infection, the immune effect and stability of the vaccine are effectively improved, and the adjuvant has a wide application prospect.

Description

Technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to an oil-in-water nanoemulsion adjuvant, and also relates to a MRSA nanoemulsion adjuvant vaccine and a preparation method of the nanoemulsion adjuvant and vaccine. Background technique [0002] With the continuous promotion of vaccine products, the application limitations caused by their natural properties are constantly exposed. Most vaccine products have a relatively short half-life in a single state, especially under the catalysis of temperature, their half-life decreases linearly with increasing temperature. Therefore, most vaccines, such as MRSA vaccines, need a suitable storage condition to maintain their stability, and the liquid environment in which they are located plays a particularly critical role in maintaining stability. The pH and ionic strength of the liquid environment and the addition of other stable components (such as some sugar molecules, amino acid components...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K9/107A61K39/085A61P31/04
Inventor 孙红武杨柳杨张卫军魏超童亚楠赵莉群冯强
Owner ARMY MEDICAL UNIV
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