Search

CN-121971597-A - AS04 adjuvant composition for vaccine, and preparation method and application thereof

CN121971597ACN 121971597 ACN121971597 ACN 121971597ACN-121971597-A

Abstract

The invention relates to the technical field of biological and pharmaceutical preparations, and discloses an AS04 adjuvant composition for vaccine, a preparation method and application thereof, wherein the composition comprises the following components of aluminum hydroxide; the preparation method comprises the steps of S1, mixing and adsorbing 3-O-deacylated-4' -monophosphoryl lipid A and aluminum hydroxide to prepare an adjuvant base core, S2, mixing and dissolving squalene and alpha-tocopherol to prepare an oil phase premix, S3, dropwise adding the oil phase premix into the adjuvant base core suspension under low shearing force, S4, adding a polymer stabilizer solution into the mixture to carry out macromolecule coating, and S5, freeze-drying the final mixture. The AS04 adjuvant is endowed with excellent freeze-drying stability, quick redissolution performance and antigen activity protection effect by sequentially constructing a multi-layer protection structure.

Inventors

  • LIU XIAOMAN
  • ZHANG QINGSHUANG
  • LIU HUAN
  • LI QI

Assignees

  • 华诺泰生物医药科技(成都)有限公司

Dates

Publication Date
20260505
Application Date
20260401

Claims (10)

  1. 1. An AS04 adjuvant composition for a vaccine, the composition comprising the following components in parts by weight: 90-110 parts of aluminum hydroxide; 10-50 parts of 3-O-deacylated-4' -monophosphoryl lipid A; 200-2000 parts of polymer stabilizer; 1-20 parts of squalene; 0.05-0.4 part of alpha-tocopherol.
  2. 2. An AS04 adjuvant composition for a vaccine according to claim 1, wherein the polymeric stabilizer is selected from poly L-histidine, poly lactic-co-glycolic acid or a combination of both.
  3. 3. An AS04 adjuvant composition for a vaccine according to claim 1, wherein the poly-L-histidine has a weight average molecular weight of 10,000 to 25,000da; In the polylactic acid-glycolic acid copolymer, the molar ratio of lactic acid to glycolic acid is 1:1, and the weight average molecular weight is 10,000-25,000 Da.
  4. 4. A process for the preparation of an AS04 adjuvant for a vaccine, characterized in that it is used for the preparation of an AS04 adjuvant composition for a vaccine according to any one of claims 1-3, comprising the steps of: S1, preparing an adjuvant-based core, namely mixing 3-O-deacylated-4' -monophosphoryl lipid A with an aluminum hydroxide gel suspension to enable the mixture to be adsorbed to obtain an adjuvant-based core suspension; s2, preparing an oil phase functional premix, namely mixing and dissolving squalene and alpha-tocopherol to obtain the oil phase functional premix; S3, sequential interface functionalization, namely dropwise adding the oil phase functional premix in the step S2 into the adjuvant-based core suspension in the step S1 under low shear force stirring; s4, polymer coating, namely dissolving the polymer stabilizer in phosphate buffer solution, and then adding the polymer stabilizer into the mixture in the step S3; And S5, freeze drying, namely freeze drying the mixture obtained in the step S4.
  5. 5. The method for preparing an AS04 adjuvant for a vaccine according to claim 4, wherein the specific operations of preparing the step S1 adjuvant-based core are AS follows: Mixing the 3-O-deacylated-4 '-monophosphoryl lipid A with an aluminum hydroxide gel suspension at a temperature of 20-28 ℃ and a stirring speed of 100-300rpm for 60-150 minutes so that the 3-O-deacylated-4' -monophosphoryl lipid A is fully adsorbed on the surface of the aluminum hydroxide particles to form a stable and uniform adjuvant-based core suspension.
  6. 6. The method of preparing AS04 adjuvant for vaccine according to claim 4, wherein in step S1, a predetermined concentration of vaccine antigen and 3-O-deacylated-4' -monophosphoryl lipid a are further added to the aluminum hydroxide gel suspension to be co-adsorbed to form an antigen-loaded adjuvant base core.
  7. 7. The method for preparing AS04 adjuvant for vaccine according to claim 4, wherein the specific operations of preparing the step S2 oil phase functional premix are: mixing the squalene with the alpha-tocopherol and continuing to stir at a stirring speed of 50-500rpm for 5-30 minutes until the alpha-tocopherol is completely dissolved, forming a uniformly clear oil phase functional premix for the subsequent steps.
  8. 8. The method for preparing AS04 adjuvant for vaccine according to claim 4, wherein the specific operations of sequential interface functionalization in step S3 are: The oil phase functional premix is dropwise added into the continuously stirred adjuvant-based core suspension at a flow rate of 0.1-1.0mL/min, the total volume of the premix accounts for 0.05% -0.3% v/v of the volume of the adjuvant-based core suspension, so that the instantaneous adsorption and spreading of the premix on the surface of the adjuvant-based core are realized, and after the dropwise addition is finished, the stirring with low shear force is continuously maintained for 15-45 minutes so as to stabilize the formed interface layer.
  9. 9. The method for preparing AS04 adjuvant for vaccine according to claim 4, wherein the specific process parameters of the step S5 of freeze-drying are: A prefreezing stage of freezing the mixture at a slab temperature of-40 ℃ to-50 ℃ for 2-4 hours, ensuring complete solidification thereof; A main drying stage, namely raising the temperature of the plate layer to-25 ℃ to-10 ℃ under the vacuum degree of 5-20Pa, and maintaining for 24-48 hours to sublimate and remove free water in the sample; and a secondary drying stage, wherein the temperature of the plate layer is further raised to 20-30 ℃ and maintained for 12-24 hours, so as to analyze and remove the bound water in the sample, and finally obtain the freeze-dried adjuvant composition.
  10. 10. Use of an AS04 adjuvant composition for a vaccine according to any one of claims 1-5 in the manufacture of a vaccine for the prevention of infectious diseases.

Description

AS04 adjuvant composition for vaccine, and preparation method and application thereof Technical Field The invention relates to the technical field of biological and pharmaceutical preparations, in particular to an AS04 adjuvant composition for a vaccine, a preparation method and application thereof. Background AS04 adjuvant is a widely used compound adjuvant composed of 3-O-deacylated-4' -monophosphoryl lipid A (MPL) adsorbed on aluminum salts such AS aluminum hydroxide. The adjuvant can synergistically activate an innate immune signal pathway and effectively induce an organism to generate a strong Th1 type cellular immune response. In order to improve the stability of vaccine products, extend their shelf life and reduce the dependency on the cold chain system, the development of liquid vaccine formulations into lyophilized dosage forms is an important technical strategy. Therefore, stable freeze-drying of vaccine compositions comprising AS04 adjuvants is achieved, with important application value. Currently, AS04 adjuvants are usually present in the form of aqueous suspensions, in which MPL molecules are adsorbed to the surface of aluminium hydroxide particles by hydrophobic and electrostatic interactions. For freeze-drying of formulations comprising biological macromolecules such as protein antigens, viral particles, etc., the prior art generally employs the addition of saccharides or polyols (e.g., trehalose, sucrose) as cryoprotectants. These protectants are capable of forming a glassy matrix of high viscosity during freezing, embedding biomolecules therein, thereby reducing to some extent the damage to ice crystal growth and dehydration processes. While prior art lyophilization with conventional cryoprotectants has been successful in certain biologicals, there are several disadvantages to applying this strategy directly to solid phase particle suspension systems such AS04 adjuvants: The method is difficult to effectively maintain the dispersibility of the adjuvant particles after freeze-drying, and often causes serious particle aggregation and re-dissolution difficulties. The reason for this is that AS04 adjuvant is a heterogeneous system with aluminium hydroxide particles AS core. During freezing, the growth of ice crystals can create a large displacement effect and mechanical stress on these micron-sized adjuvant particles, forcing intimate contact between the particles. Conventional saccharide protectants, while capable of forming a macroscopic glassy state, do not form an effective physical barrier on the surface of each individual adjuvant particle. When moisture is removed by sublimation, solid bridges, which are difficult to break by simple re-dissolution, are formed between particle surfaces lacking hydration layers due to van der Waals forces, electrostatic attraction, and the like, resulting in irreversible aggregation. Furthermore, this method has limited protective effect on the biological activity of the antigen adsorbed on the adjuvant surface. The protein antigen is immobilized on the surface of aluminum hydroxide through physical adsorption, and the solid-liquid interface is a high-energy unstable area. During lyophilization and dehydration, antigen molecules undergo interfacial water molecule structure changes and dehydration stress, which are very likely to cause changes in their spatial conformation or even denaturation. Conventional protectants are distributed throughout the solution system and cannot provide targeted protection in the local microenvironment where the adjuvant particles are bound to the antigen, thus it is difficult to effectively prevent interface-induced antigen inactivation, resulting in reduced immunogenicity of the vaccine product after reconstitution. Finally, the lyophilized product thus obtained has poor long-term storage stability. The problem of aggregation among particles cannot be fundamentally solved, and a large number of defects and stresses generated by aggregation of particles exist in the formed freeze-dried matrix. Such unstable structures are prone to further physical changes (such as matrix collapse) during long term storage, particularly under temperature fluctuations, accelerating solid state aggregation of the particles and slow degradation of the antigen, thus shortening the effective shelf life of the product. Disclosure of Invention The invention aims to provide an AS04 adjuvant composition for vaccine, a preparation method and application thereof, and solves the problems that AS04 adjuvant is easy to generate irreversible aggregation in the freeze drying process, and is difficult to redissolve and the activity of loaded antigen is reduced. In order to achieve the above purpose, the invention is realized by the following technical scheme: the invention provides an AS04 adjuvant composition for vaccine, which adopts the following technical scheme: an AS04 adjuvant composition for a vaccine, the composition comprising the following component