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CN-122005502-A - Antigen double-burst self-boosting immunity vaccine and application thereof

CN122005502ACN 122005502 ACN122005502 ACN 122005502ACN-122005502-A

Abstract

The invention relates to the technical field of medicine, in particular to an antigen double-burst self-boosting immune vaccine and application thereof, wherein antigen is loaded in the interior and the exterior of a complex coacervation microcapsule in the vaccine, the vaccine comprises nanoparticle prepared by a nano precipitation method, functionalized nanoparticle after the nanoparticle and cell membrane are coated with antigen together, the micro-fluidic technology is used for coating the functionalized nano-particles by a complex coacervation method to prepare the micro-capsules, the self-polymerization reaction of dopamine hydrochloride is utilized to coat the surface of the micro-capsules as coating micro-capsules, and finally the micro-capsules are blended with the functionalized nano-particles, so that antigens are separated into an inner part and an outer part. The prepared vaccine can release antigen firstly, then effectively recruit and activate antigen presenting cells, be absorbed by the antigen presenting cells to cause immune response, and can effectively induce the generation of specific antibodies by realizing the release of the antigen twice through a time-lapse coating so as to simulate the traditional natural immune procedure.

Inventors

  • GAO JING
  • ZHU LIN
  • ZHANG WENPENG
  • LI ZHIPING
  • RAN YUHUA
  • ZHAI YANAN

Assignees

  • 中国人民解放军军事科学院军事医学研究院

Dates

Publication Date
20260512
Application Date
20260203

Claims (10)

  1. 1. A double burst release pulsatile release vaccine, characterized in that the vaccine is a poly-dopamine coated complex coacervated microcapsule, both inside and outside of which carry antigens.
  2. 2. The double burst release pulsed vaccine of claim 1, wherein the antigen is selected from the group consisting of a spotag-OVA antigen, a chicken ovalbumin antigen, a recombinant antigen, a novel coronavirus antigen, a respiratory syncytial virus antigen, an influenza virus antigen, and a hepatitis b virus antigen, and wherein the nucleotide sequence of the spotag-OVA antigen is shown in SEQ ID No. 1.
  3. 3. The dual burst release pulsatile vaccine of claim 1, further comprising an adjuvant selected from the group consisting of an aluminum adjuvant selected from the group consisting of aluminum hydroxide gel, aluminum phosphate, aluminum sulfate, ammonium alum, or potassium alum, or a small molecule adjuvant selected from the group consisting of CpG, toll-like receptor agonists, or squalene.
  4. 4. The dual burst release pulsed vaccine of claim 1, wherein the particle size of the complex coacervation microcapsule is 10-150 μm and the particle size of the nanoparticle is 20-500nm.
  5. 5. A method of preparing a double burst pulsed release vaccine according to any one of claims 1 to 4, comprising the steps of: (1) Preparing nanoparticles by adopting a nano precipitation method; (2) The nanoparticle and a cell membrane are coated with fusion protein antigen together to obtain a functionalized nanoparticle; (3) Coating the functionalized nanoparticles and the adjuvant by a complex coacervation method by utilizing a microfluidic technology to prepare a complex coacervation microcapsule; (4) And (3) performing polydopamine coating on the surface of the complex coacervation microcapsule, and blending with the functionalized nanoparticles to obtain the double-burst release pulse release vaccine.
  6. 6. The method according to claim 5, wherein in the step (1), the nano-precipitation method uses an organic solvent to dissolve the high molecular polymer to prepare an oil phase, and uses a surfactant to prepare a solution as an aqueous phase; The high polymer is selected from polylactic acid, polyanhydride, lactide-glycolide copolymer or polycaprolactone, the organic solvent is selected from dichloromethane, ethyl acetate, chloroform, acetone, diethyl ether or ethanol, and the surfactant is selected from polyethylene glycol octyl phenyl ether, tween, polyoxyethylene, polyvinyl alcohol, span, poloxamer, polyethylene glycol or sodium dodecyl sulfate.
  7. 7. The preparation method of claim 5, wherein in the step (3), the three-phase air pressure of the microfluidic technology is 10-2000mpa, the internal phase of the complex coacervation microcapsule is an antigen and adjuvant solution, the intermediate phase is a capsule material solution, the capsule material is selected from gelatin, acacia or sodium carboxymethyl cellulose, and the external phase is fluorinated oil added with a surfactant.
  8. 8. The method according to claim 5, wherein in the step (4), the polydopamine coating is performed in a neutral buffer having a pH of 6 to 8 for a period of 1 to 240 minutes, and the neutral buffer is selected from PBS, sodium citrate aqueous solution or sodium dihydrogen phosphate aqueous solution.
  9. 9. Use of a double burst pulsed release vaccine according to any one of claims 1-4 in the preparation of a prophylactic or therapeutic vaccine.
  10. 10. The use according to claim 9, wherein the prophylactic or therapeutic vaccine is for the prevention or treatment of novel coronavirus infection, respiratory syncytial virus infection, influenza or hepatitis b, and wherein the vaccine is administered by intramuscular injection, subcutaneous injection or pulmonary administration.

Description

Antigen double-burst self-boosting immunity vaccine and application thereof Technical Field The invention relates to the technical field of medicines, in particular to an antigen double-burst self-boosting immune vaccine and application thereof. Background The infection is one of the main factors threatening global public health and human life safety for a long time, the outbreak of new epidemic situation further highlights the importance of an infection prevention system, and the vaccination is the most effective means for coping with biological safety threat and blocking the transmission of the infection, and the technical innovation and optimization are always the research focus of the medicine field. According to the classification of world health organization, the vaccines clinically applied at present mainly comprise six major types of attenuated live vaccines, inactivated vaccines, antitoxin, subunit vaccines, vector vaccines, nucleic acid vaccines and the like. The subunit vaccine only contains specific antigen components of pathogens, does not contain nucleic acid substances, has the obvious advantages of no infectious risk, high safety, strong immune pertinence and the like, and has wide application prospect in infectious disease prevention. However, the core short plate of subunit vaccines is poorly immunogenic, and it is difficult to elicit a long-lasting and effective immune protective response in the body alone, and it is often necessary to boost the immune effect by booster vaccination after priming (i.e., a multi-needle immunization program) to achieve establishment of long-term immune memory. Although the multi-needle immunization program is a main mode for making up the insufficient immunogenicity of subunit vaccines currently, the method has a plurality of limitations which are difficult to overcome in practical application, wherein ① is easy to cause immunization loss, the multi-needle immunization program is long in time consumption and high in inoculation cost, a perfect inoculation management system is needed to support, partial people are difficult to complete whole-course inoculation on time under the condition of underdeveloped economic areas or limited inoculation resources, so that the coverage rate of immune protection is influenced, ② special people are poor in inoculation compliance, the mobility of people with standing personnel in the border, field operators and the like or the special groups in the working environment are difficult to strictly follow a preset schedule to inoculate, so that the immunization program is interrupted, ③ is insufficient in adaptability in sudden major epidemic situations, a large amount of social resources are needed to be mobilized for vaccine production, transportation and inoculation service, the cost is high, and the group immune barrier can not be constructed quickly due to vaccine supply shortage, and the availability of the vaccine is particularly limited in developing countries due to excessive inoculation times. Therefore, it is a key to solve the above problems to develop a vaccine formulation capable of achieving a single-needle vaccination, i.e., achieving a multi-needle-like immune effect. The single-needle vaccine not only needs to have long-acting immunity protection capability, but also needs to be compatible with vaccination convenience so as to improve vaccination coverage rate and reduce vaccination cost, and simultaneously meets the requirement of quick prevention and control under sudden epidemic situation. The method comprehensively considers the necessity of enhancing immunity and placing immune loss, develops a single-needle double-burst release vaccine which can realize antigen pulse release and simulate natural immunization program in vivo, and has important practical significance and application value for breaking through the application bottleneck of the existing subunit vaccine and improving the infectious disease prevention and control efficiency. Disclosure of Invention The invention aims to provide a nanoparticle vaccine for realizing in vivo self-boosting immunity through antigen pulse release, antigen proteins are loaded into microcapsules, time-lag intervals can be realized, the core pain point of the traditional subunit vaccine which needs multi-needle immunity is effectively solved, and the nanoparticle vaccine can be used for immunizing organisms through various administration modes such as intramuscular injection, subcutaneous injection, pulmonary inhalation and the like, and inducing in vivo immune response. The invention aims at realizing the following technical scheme: The invention provides a double-burst release pulse release vaccine, which is a complex coacervation microcapsule with a polydopamine coating, wherein the inside and the outside of the complex coacervation microcapsule are loaded with antigens. Further, the antigen is selected from a Spytag-OVA antigen, a chicken ovalbumin antigen, a recomb