Novel retinoic acid nanoemulsion adjuvant for efficiently enhancing humoral immune response and mucosal immune response and preparation method and application of adjuvant

A technology of retinoic acid and nanoemulsion, which is applied in the field of biomedicine to achieve the effects of complete structure, enhanced immune response, and good uniformity

Pending Publication Date: 2021-11-30
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nanoemulsions can greatly improve the solubility of insoluble drugs, promote drug absorption, enhance drug stability, and improve bioavailability, but there are no reports of retinoic acid nanoemulsion adjuvants at home and abroad

Method used

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  • Novel retinoic acid nanoemulsion adjuvant for efficiently enhancing humoral immune response and mucosal immune response and preparation method and application of adjuvant
  • Novel retinoic acid nanoemulsion adjuvant for efficiently enhancing humoral immune response and mucosal immune response and preparation method and application of adjuvant
  • Novel retinoic acid nanoemulsion adjuvant for efficiently enhancing humoral immune response and mucosal immune response and preparation method and application of adjuvant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1, formulation screening of retinoic acid nanoemulsion adjuvant (RA-NE)

[0046] Formula 1: Retinoic acid nanoemulsion adjuvant (100 g): 1 g of retinoic acid, 2 g of Tween-80, 2 g of propylene glycol, and 95 g of distilled water.

[0047] Recipe 2: Retinoic acid nanoemulsion adjuvant (100 g): 0.1 g of retinoic acid, 30 g of Tween-80, 10 g of glycerin, 10 g of glyceryl caprylate (GTCC), and 49.9 g of distilled water.

[0048] Formula 3: Retinoic acid nanoemulsion adjuvant (100 g): 2 g of retinoic acid, 5 g of Tween-20, 0.1 g of propylene glycol, 0.1 g of ethyl acetate, and 92.8 g of distilled water.

[0049] Formula 4: retinoic acid nanoemulsion adjuvant (100g): retinoic acid 5g, Tween-60 20g, propylene glycol 4g, medicinal paraffin oil 6g, distilled water 65g.

[0050] Formula 5: retinoic acid nanoemulsion adjuvant (100g): 2g retinoic acid, 30g Tween-85, 10g 1,3-butanediol, 15g GTCC, 43g distilled water.

[0051] Formula 6: retinoic acid nanoemulsion adjuva...

Embodiment 2

[0072] Example 2, Preparation and Characterization of Preferred atRA-NE

[0073] 1. Preparation of atRA-NE

[0074] Accurately weigh 10 mg of retinoic acid (atRA) in a 50 ml clean and dry beaker, drop the oil phase, surfactant and co-surfactant and stir until fully dissolved, then adopt the low-energy emulsification method, that is, slowly add sterilized pure Water until the volume of the system is 10mL, prepare retinoic acid nanoemulsion adjuvant (atRA-NE), and store at room temperature in the dark. Blank nanoemulsion adjuvant (BNE), there is no need to add retinoic acid in the prescription.

[0075] 2. Particle size and Zeta potential detection

[0076] The prepared nanoemulsion adjuvant was diluted 100 times with ultrapure water, and the particle size and Zeta potential were detected in a Nano ZA dynamic light scattering particle size potentiometer.

[0077] 3. Transmission electron microscope (TEM) detection

[0078] Dilute the prepared nanoemulsion adjuvant 100 times ...

Embodiment 3

[0088] Example 3, atRA-NE stimulates mouse lymphocyte maturation ability

[0089] 1. The ability of atRA-NE to promote the maturation of BMDC cells

[0090]Kill the mouse and take its femur, use a 1mL syringe to draw 1640 medium containing 10% FBS to wash the bone marrow 5 times, collect the bone marrow cell washing solution, centrifuge at 1500rpm for 15 minutes, and plant the collected cells in the cell culture plate, every other day Half the amount of medium was changed once, and the cells were collected by centrifugation on the 6th day to obtain mouse bone marrow-derived dendritic cells (BMDCs), and the cell concentration was adjusted to 5×10 5 pc / mL, add 2ml / well in a 6-well plate. Afterwards, 20 μL medium, 20 μL 1:100 diluted Blank-NE, 20 μg atRA or 20 μL 1:100 diluted atRA-NE were added to each well, mixed by blowing and placed in a cell culture incubator for 24 hours. Centrifuge and wash cells with PBS, dilute APC-labeled mouse CD11c antibody, PE-labeled mouse CD40 an...

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Abstract

The invention discloses a novel retinoic acid nanoemulsion adjuvant for efficiently enhancing humoral immune response and mucosal immune response and a preparation method and application of the adjuvant. The retinoic acid nanoemulsion adjuvant is prepared from retinoic acid, an oil phase, a surfactant, a cosurfactant and a water phase, wherein the retinoic acid is wrapped by a formed oil-in-water type nanoemulsion adjuvant system. The retinoic acid nanoemulsion adjuvant can be delivered together with multiple antigens (a model antigen OVA, a staphylococcus aureus recombinant protein antigen and a neocoronavirus recombinant protein antigen) respectively, the immune response level of a vaccine antigen can be enhanced after injection immunization, the adjuvant has a good immune protection effect, can induce humoral immune response of a system and efficiently activate immune response of mucosal parts (intestinal mucosa, vaginal mucosa, lung mucosa, gastric mucosa and nasal mucosa) and has great market application value and broad application prospect.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a novel retinoic acid nano-emulsion adjuvant for efficiently enhancing humoral immune response and mucosal immune response, a preparation method and application thereof. Background technique [0002] As the first line of defense to block the invasion of pathogens, mucous membranes are widely distributed in the respiratory, digestive, urinary, and reproductive systems of the human body, and play an irreplaceable role in the process of protecting human health. If the mucous membrane cannot effectively block pathogens, its moist and fragile structure will become an ideal breeding ground for pathogens, and many important organs will be directly exposed to the threat of pathogens. Pathogens that infect the human body from mucous membranes, such as Vibrio cholerae, Salmonella, and rotavirus, have always seriously threatened human health. Vaccination is an important means to preven...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61P37/04
CPCA61K39/39A61P37/04A61K2039/55566A61K2039/55511A61K2039/575A61K2039/57
Inventor 李海波孙红武金喆邹全明冒灵刘畅刘婧怡曾浩薛若怡冯壤李郭成金滋力
Owner ARMY MEDICAL UNIV
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