Search

CN-121987775-A - Preparation method and application of novel hydrogel immunopotentiator

CN121987775ACN 121987775 ACN121987775 ACN 121987775ACN-121987775-A

Abstract

The invention discloses a preparation method and application of a novel hydrogel immunopotentiator, and belongs to the technical field of veterinary medicine. The preparation method comprises the steps of adding ferric trichloride solution into hyaluronic acid solution, reacting under a stirring state, then adjusting pH to 5.5-7.5, stirring to obtain hyaluronic acid hydrogel, and adding 2',3' -cGAMP with the final concentration of 0.1-800 mug/mL into the hyaluronic acid hydrogel to obtain the hydrogel immunopotentiator. The invention also discloses application of the hydrogel immunopotentiator in aluminum gel inactivated vaccine. The immunopotentiator has a 3D network structure, can efficiently and targetedly deliver an agonist to drainage lymph nodes, avoids systemic inflammatory response and better plays a role in immunopotentiation, is used for aluminum gel inactivated vaccine for porcine epidemic diarrhea, can remarkably up-regulate serum neutralizing antibody level, can effectively induce Th1 type cell immune response, and provides stronger immunoprotection response.

Inventors

  • LI LAN
  • WANG YIWEI
  • ZHENG QISHENG
  • CHEN Jin
  • QIAO XUWEN
  • QIN ZHU
  • HOU LITING
  • ZHANG YUANPENG
  • DU LUPING
  • YU XIAOMING

Assignees

  • 江苏省农业科学院

Dates

Publication Date
20260508
Application Date
20260213

Claims (10)

  1. 1. A preparation method of the novel hydrogel immunopotentiator is characterized by comprising the steps of adding ferric trichloride solution into hyaluronic acid solution, reacting in a stirring state, then adjusting pH to 5.5-7.5, stirring to obtain hyaluronic acid hydrogel, and adding 2',3' -cGAMP with the final concentration of 0.1-800 mug/mL into the hyaluronic acid hydrogel to obtain the novel hydrogel immunopotentiator.
  2. 2. The preparation method according to claim 1, wherein the concentration of the ferric trichloride solution is 8-60mg/mL, the concentration of the hyaluronic acid solution is 8-12mg/mL, and the concentration of 2',3' -cGAMP in the novel hydrogel immunopotentiator is 50-200 μg/mL.
  3. 3. The preparation method according to claim 1 or 2, wherein the reaction time of the hyaluronic acid solution and the ferric trichloride solution is 30min to 6h, preferably 1 to 3h, and the stirring time after the pH is adjusted to 5.5 to 7.5 is 1.5 to 4h.
  4. 4. The preparation method of the sodium hyaluronate, as set forth in claim 3, characterized in that the ferric trichloride solution is obtained by dissolving ferric trichloride hexahydrate in a hydrochloric acid solution, the hyaluronic acid solution is obtained by dissolving sodium hyaluronate with a molecular weight of 400-1000 kDa in physiological saline, and the pH is adjusted by adopting an alkaline solution, wherein the alkaline solution is an aqueous NaOH solution with a molar concentration of 0.01-2 mM.
  5. 5. The preparation method according to claim 4, wherein 2',3' -cGAMP is dissolved in purified water to prepare a 1-3mg/mL solution, and then added to hyaluronic acid hydrogel.
  6. 6. The novel hydrogel immunopotentiator prepared by the method of any one of claims 1 to 5.
  7. 7. An aluminum gel inactivated vaccine for porcine epidemic diarrhea comprising the novel hydrogel immunopotentiator of claim 6.
  8. 8. Vaccine according to claim 7, characterized in that the aluminium gel is aluminium hydroxide or aluminium phosphate, preferably aluminium hydroxide.
  9. 9. The preparation method of the vaccine of claim 7, which is characterized by comprising the steps of adding aluminum gel into inactivated porcine epidemic diarrhea virus liquid, stirring to obtain an inactivated vaccine, dripping the novel hydrogel immunopotentiator into the inactivated vaccine, and stirring to obtain the vaccine.
  10. 10. Vaccine according to claim 9, characterized in that the volume ratio of the novel hydrogel immunopotentiator to the inactivated vaccine is 1:1-99, preferably 1:1-19.

Description

Preparation method and application of novel hydrogel immunopotentiator Technical Field The invention belongs to the technical field of biotechnology and vaccine, and particularly relates to a preparation method and application of a novel hydrogel immunopotentiator. Background Cyclic dinucleotides (cyclic dinucleotides, CDNs) are agonists of the cyclic guanosine-monophosphate synthase-interferon gene stimulatory factor (cGAS-STING) natural immune signaling pathway, which has been attracting attention in recent years due to its key role in innate immune activation, recognizes abnormal DNA through cGAS, catalyzes the synthesis of cGAMP, binds to the V-shaped dimer pocket formed by the cytoplasmic domain of STING proteins as a second messenger, and after undergoing conformational changes, depolymerizes STING proteins from the endoplasmic reticulum to golgi and lysosomes, recruits TBK1 and IRF3, activates the type I interferon pathway, enhances the immune response of the organism, providing a new direction for vaccine development. CDNs is taken as a small molecular compound, is very easy to enter blood circulation, cannot be effectively transported to lymph nodes to play an immunopotentiating role, even induces systemic inflammatory response, causes immune side effects, has a phosphodiester bond at CDNs, is very easy to degrade by external nucleotide pyrophosphatase/phosphodiesterase 1 (ENPP 1), has a short half-life period, and CDNs molecules are negatively charged, have water solubility, are very easy to diffuse in vivo and are difficult to penetrate cell membranes, so that the practical application of CDNs is severely restricted. In view of this, a great deal of research is focused on developing new delivery systems suitable for CDNs, such as nanoparticles, cationic liposomes, engineered exosomes, smart hydrogels, etc., but there are drawbacks of cumbersome preparation, low drug loading rate, high cost, etc., resulting in CDNs not being widely used clinically. Disclosure of Invention The invention aims to provide a preparation method of a novel hydrogel immunopotentiator, which is simple and has high drug loading rate, and the obtained hydrogel immunopotentiator can quickly excite immune response, can permanently maintain antibody level and provides long-term and stable protection for organisms. The invention adopts the following technical scheme: A preparation method of the novel hydrogel immunopotentiator comprises the following steps of adding ferric trichloride solution into hyaluronic acid solution, reacting under a stirring state, then adjusting pH to 5.5-7.5, stirring to obtain hyaluronic acid hydrogel, and adding 2',3' -cGAMP with the final concentration of 0.1-800 mug/mL into the hyaluronic acid hydrogel to obtain the novel hydrogel immunopotentiator. In the invention, the concentration of the ferric trichloride solution is 8-60mg/mL, the concentration of the hyaluronic acid solution is 8-12mg/mL, and the concentration of the 2',3' -cGAMP in the novel hydrogel immunopotentiator is 50-200 mug/mL. In the invention, the reaction time of the hyaluronic acid solution and the ferric trichloride solution is 30 min-6 h, preferably 1-3 h, and the stirring time after the pH is adjusted to 5.5-7.5 is 1.5-4h. The ferric trichloride solution is prepared by dissolving ferric trichloride hexahydrate in hydrochloric acid solution, the hyaluronic acid solution is prepared by dissolving sodium hyaluronate with molecular weight of 400-1000 kDa in physiological saline, and the pH is regulated by adopting alkaline solution, wherein the alkaline solution is NaOH aqueous solution with molar concentration of 0.01-2 mM. In the present invention, 2',3' -cGAMP was dissolved in purified water to prepare a 1-3mg/mL solution, which was then added to hyaluronic acid hydrogel. The invention also provides a novel hydrogel immunopotentiator prepared by the method. The invention also provides an aluminum gel inactivated vaccine for porcine epidemic diarrhea containing the novel hydrogel immunopotentiator. In the present invention, the aluminum paste is aluminum hydroxide or aluminum phosphate, preferably aluminum hydroxide. The invention also provides a preparation method of the vaccine, which comprises the following steps of adding aluminum gel into an inactivated porcine epidemic diarrhea virus liquid, stirring to obtain an inactivated vaccine, dripping the novel hydrogel immunopotentiator into the inactivated vaccine, and stirring to obtain the vaccine. In the preferred technical scheme, the volume ratio of the novel hydrogel immunopotentiator to the inactivated vaccine is 1:1-99, preferably 1:1-19. Compared with the prior art, the method has the beneficial effects that the hydrogel slow release platform with the 3D network structure is adopted to wrap the natural immune agonist, the method is simple, the drug loading rate is high, the slow release effect of the hydrogel is cooperated with the effect of the glycan targeting lymph n