CN-122005778-A - Herpes zoster vaccine containing novel adjuvant and preparation method thereof

CN122005778ACN 122005778 ACN122005778 ACN 122005778ACN-122005778-A

Abstract

The invention discloses a herpes zoster vaccine containing a novel adjuvant and a preparation method thereof, and relates to the technical field of vaccines. The vaccine is prepared by using gE protein as an antigen, compounding liposome and PLGA nano particles, preparing liposome from phospholipid, cholesterol and cholesterol succinic acid monoester, mixing the liposome with an antigen solution containing a film forming agent, performing ultrasonic treatment, pre-freezing and freeze-drying to obtain a re-soluble liposome freeze-dried preparation, using PLGA as a carrier, preparing nanoparticle dispersion liquid for encapsulating immune enhancement factors through emulsification, dialysis and high-pressure homogenization, mixing the two, and performing freeze-drying again after stabilizing the mixture by using polyvinyl alcohol. The problems of aggregation, particle size fluctuation and antigen conformation damage caused by easy oxidative destabilization of liposome-antigen freeze-dried preparation in freeze-drying, re-dissolution and storage are solved, and the re-dissolution stability and consistency are improved through an internal and external synergistic antioxidant stabilizing system.

Inventors

WANG HAILONG
LI JUN
QI SHUYA
ZHAO BIN

Assignees

江苏华诺泰生物医药科技有限公司

Dates

Publication Date: 20260512
Application Date: 20260409

Claims (10)

1. A novel adjuvant-containing herpes zoster vaccine is characterized by being a freeze-dried preparation, comprising a liposome freeze-dried preparation containing a herpes zoster virus antigen and PLGA nanoparticle dispersion liquid loaded with an immune enhancement factor, wherein the liposome freeze-dried preparation is re-dissolved and then mixed with the PLGA nanoparticle dispersion liquid, and the novel adjuvant-containing herpes zoster vaccine freeze-dried preparation is obtained through freeze drying.
2. The novel adjuvanted herpes zoster vaccine of claim 1 wherein the lyophilized preparation of liposomes comprising the herpes zoster virus antigen is comprised of the herpes zoster virus antigen, phospholipids, cholesterol and cholesterol succinate monoester.
3. The novel adjuvant-containing herpesvaccine of claim 2, wherein the phospholipid is at least one of soybean lecithin, hydrogenated lecithin, egg yolk lecithin, phosphatidylcholine, phosphatidylserine, distearoyl phosphatidylcholine, dipalmitoyl phosphatidylcholine, or dimyristoyl phosphatidylcholine.
4. The herpes zoster vaccine with novel adjuvant according to claim 1 further comprising an auxiliary material in a liposome lyophilized preparation containing the herpes zoster virus antigen, wherein the auxiliary material is at least one of PEGylated lipid and tocopherol.
5. The herpesvaccine with novel adjuvant of claim 4, wherein the PEGylated lipid is at least one of DSPE-PEG2000, DSPE-PEG-MALEIMIDE, DSPE-PEG-Biotin, DSPE-PEG-Mannose or DSPE-PEG-glutathione, and the tocopherol is tocopheryl succinate.
6. The herpes zoster vaccine with novel adjuvant according to claim 1 characterized in that the immune enhancing factor in the PLGA nanoparticle lyophilized preparation loaded with the immune enhancing factor is at least one of a TLR agonist, a STING agonist, an oligodeoxynucleotide immunomodulator, a saponin immunopotentiator, a polysaccharide immunopotentiator or a cytokine immunomodulator.
7. A method of preparing a novel adjuvant-containing shingles vaccine according to any of claims 1-6, comprising the steps of: Step 1, adding phospholipid, cholesterol and cholesterol succinic acid monoester into a mixed solvent, stirring uniformly, transferring to a rotary evaporator to remove the solvent to obtain a film-shaped substance, adding the film-shaped substance into water, mixing uniformly, heating to obtain a liposome suspension, adding gE protein and a film-forming agent into water, mixing uniformly to obtain an antigen solution, adding the antigen solution into the liposome suspension, mixing uniformly, performing ultrasonic treatment, and finally placing the mixed system into pre-freezing, and freeze-drying to obtain a liposome freeze-dried preparation; Step 2, adding PLGA into dichloromethane, mixing uniformly to obtain an oil phase, adding immune enhancement factors and PVA into water, mixing uniformly to obtain a water phase, adding the oil phase into the water phase, stirring to obtain emulsion, transferring to a rotary evaporator to remove solvent, transferring to a dialysis bag for dialysis, and homogenizing by using a high-pressure homogenizer to obtain nanoparticle dispersion liquid; And step 3, adding the prepared liposome freeze-dried preparation into PBS buffer solution, uniformly mixing, adjusting the gE protein concentration in the liposome, adjusting the nanoparticle dispersion liquid according to the PLGA mass concentration, uniformly mixing the liposome solution and the nanoparticle dispersion liquid, then adding PVA, continuously stirring uniformly, pre-freezing, and freeze-drying to obtain the herpes zoster vaccine containing the novel adjuvant.
8. The method for preparing a novel adjuvant-containing herpesvaccine according to claim 7, wherein the mixed solvent is prepared by mixing chloroform and methanol in a volume ratio of (1-3): 1.
9. The method of preparing a novel adjuvant-containing shingles vaccine according to claim 7, wherein the film-forming agent is at least one of proline, hydroxypropyl-beta-cyclodextrin, sulfobutyl ether-beta-cyclodextrin, hydroxypropyl methylcellulose, dextran, sodium caseinate, or gelatin.
10. The method for preparing the herpes zoster vaccine with the novel adjuvant according to claim 7, wherein the ultrasonic treatment condition in the step (1) is that the ultrasonic power is 80-100W, the working time is 1-5s, the intermittent time is 1-5s, the total ultrasonic time is 10-20min, the prefreezing condition is-60 ℃ to-80 ℃ and the time is 1-3h, and the freeze drying condition is-20 ℃ to-40 ℃ and the time is 0.1-0.3mbar and the time is 12-24h.

Description

Herpes zoster vaccine containing novel adjuvant and preparation method thereof Technical Field The invention relates to the technical field of vaccines, in particular to a herpes zoster vaccine containing a novel adjuvant and a preparation method thereof. Background Herpes zoster (herps zoter) is caused by latent reactivation of varicella-zoster virus (VZV), and the risk of onset increases with age and with reduced immune function. The core of the existing prevention is that the specific immunity of the VZV is enhanced by a vaccine in the adult stage, and the complication risks such as the onset of the disease and the neuralgia after the herpes zoster are reduced. The existing technical route of the herpes zoster vaccine mainly comprises two types, namely an attenuated live vaccine, inducing immunity through a preparation containing the attenuated live virus, and a recombinant subunit vaccine, wherein glycoprotein E (gE) of VZV is used as a key antigen and is matched with a strong adjuvant system so as to improve the intensity and the persistence of immune response of the aged and the like. CN108992667A discloses a herpes zoster vaccine, a preparation method and application thereof, and aims at solving the problem that the existing herpes zoster vaccine has higher safety risk, wherein an adjuvant used by the herpes zoster vaccine is Poly (I: C). Poly (I: C) is abbreviated as Poly inosinic acid, double-stranded RNA, and the safety of the Poly (I: C) is proved in large-scale clinical use. Poly (I: C) is a potent interferon inducer that can produce an immune response in vivo resembling a viral infection, and can induce CD4 + and CD8 + T cells, enhancing cellular and humoral immune responses. However, they tend to be oxidatively unstable in storage and to cause aggregation and particle size fluctuations. Disclosure of Invention In view of the above-mentioned drawbacks of the prior art, the present invention aims to solve the technical problem of improving aggregation, particle size fluctuation and antigen conformation damage caused by easy oxidative destabilization of liposome-antigen freeze-dried preparation in freeze-drying, reconstitution and storage, and to improve reconstitution stability and consistency by an internal and external synergistic antioxidant stabilizing system. Liposomes and nanoparticle complexes can be used to deliver a variety of antigens, particularly in vaccines, where the liposomes act as antigen carriers, are effective to encapsulate the antigen and provide slow release. The nanoparticle can be used as adjuvant to promote antigen presenting cell uptake and enhance immune response. In order to achieve the above purpose, the invention provides a herpes zoster vaccine containing a novel adjuvant and a preparation method thereof. A method for preparing a herpes zoster vaccine containing a novel adjuvant, comprising the following steps: Step 1, adding phospholipid, cholesterol and cholesterol succinic acid monoester into a mixed solvent, stirring uniformly, transferring to a rotary evaporator to remove the solvent to obtain a film-shaped substance, adding the film-shaped substance into water, mixing uniformly, heating to obtain a liposome suspension, adding gE protein and a film-forming agent into water, mixing uniformly to obtain an antigen solution, adding the antigen solution into the liposome suspension, mixing uniformly, performing ultrasonic treatment, and finally placing the mixed system into pre-freezing, and freeze-drying to obtain a liposome freeze-dried preparation; Step 2, adding PLGA into dichloromethane, mixing uniformly to obtain an oil phase, adding immune enhancement factors and PVA into water, mixing uniformly to obtain a water phase, adding the oil phase into the water phase, stirring to obtain emulsion, transferring to a rotary evaporator to remove solvent, transferring to a dialysis bag for dialysis, and homogenizing by using a high-pressure homogenizer to obtain nanoparticle dispersion liquid; And 3, adding the prepared liposome freeze-dried preparation into PBS buffer solution, uniformly mixing, adjusting the gE protein concentration in the liposome, adjusting the nanoparticle dispersion liquid according to the PLGA mass concentration, uniformly mixing the liposome solution and the nanoparticle dispersion liquid according to the volume ratio of 1:1, adding PVA, continuously stirring uniformly, pre-freezing, and freeze-drying to obtain the herpes zoster vaccine containing the novel adjuvant. Wherein the phospholipid is at least one of soybean lecithin, hydrogenated lecithin, egg yolk lecithin, phosphatidylcholine, phosphatidylserine, distearoyl phosphatidylcholine (DSPC), dipalmitoyl phosphatidylcholine (DPPC), and dimyristoyl phosphatidylcholine (DMPC); wherein the mixed solvent is formed by mixing chloroform and methanol according to the volume ratio of (1-3): 1; wherein the film forming agent is at least one of proline, hydroxypropyl-beta-cyclodextrin, sulfobutyl et