阅读说明：本技术 一种热稳定的甲肝疫苗可溶性微针贴及其制备方法 (Thermostable hepatitis A vaccine soluble microneedle patch and preparation method thereof ) 是由 张庶民 马凤森 周荔葆 廖辉 陈中秋 刘苗苗 吴铮 辛小韵 于 2018-07-04 设计创作，主要内容包括：本发明提供了一种热稳定的甲肝疫苗可溶性微针贴,包括针体和背衬,所述的针体由甲肝疫苗、基质材料和疫苗稳定剂组成,其中,所述的疫苗稳定剂为聚乙烯吡咯烷酮和右旋糖酐的混合,所述的疫苗稳定剂与甲肝疫苗的含量比为(2：1)～(5：1)。本发明制备的甲肝疫苗可溶性微针贴,针体药物热稳定性好,机械强度适中,能有效刺入皮肤。(The invention provides a thermostable hepatitis A vaccine soluble microneedle patch, which comprises a needle body and a back lining, wherein the needle body consists of a hepatitis A vaccine, a matrix material and a vaccine stabilizer, the vaccine stabilizer is a mixture of polyvinylpyrrolidone and dextran, and the content ratio of the vaccine stabilizer to the hepatitis A vaccine is (2:1) - (5: 1). The soluble microneedle patch for the hepatitis A vaccine prepared by the invention has good heat stability of the drug on the needle body and moderate mechanical strength, and can effectively penetrate into the skin.)

1. A heat-stable hepatitis A vaccine soluble microneedle patch comprises a needle body and a back lining, and is characterized in that the needle body consists of a hepatitis A vaccine, a matrix material and a vaccine stabilizer, wherein the vaccine stabilizer is a mixture of polyvinylpyrrolidone and dextran, and the content ratio of the vaccine stabilizer to the hepatitis A vaccine is (2:1) - (5: 1).

2. The heat-stable hepatitis A vaccine soluble microneedle patch according to claim 1, wherein the content ratio of polyvinylpyrrolidone to dextran in said vaccine stabilizer is (3:1) - (1: 3).

3. The heat-stable hepatitis A vaccine dissolvable microneedle patch according to claim 1, wherein said matrix material is polyvinyl alcohol.

4. The vaccine stabilizer according to claim 1, wherein the content ratio of polyvinylpyrrolidone to dextran is 2: 1.

5. the heat-stable hepatitis A vaccine soluble microneedle patch according to claims 1-4, wherein the needle body comprises the following components in percentage by weight: 30 wt% of polyvinyl alcohol, 37.3 wt% of polyvinylpyrrolidone, 18.7 wt% of dextran and 14 wt% of hepatitis A vaccine.

6. The method for preparing a thermostable hepatitis a vaccine dissolvable microneedle patch according to claim 1, comprising the steps of:

(1) negative mold preparation of microneedles

Firstly, preparing male mold microneedles by adopting photoetching and etching processes, then mixing polysiloxane and a curing agent according to the mass ratio of 6:1, and pouring the mixture into a cuboid container in which the monocrystalline silicon male mold microneedles are placed; placing the container in a vacuum drying box, and vacuumizing to remove bubbles in the mixed solution; then putting the molded polysiloxane into an oven, drying for 3 hours, and taking out to obtain a molded polysiloxane mold;

(2) first vacuum mold

Proportionally mixing hepatitis A vaccine, matrix material and vaccine stabilizer in a prescription amount, adding a small amount of eosin aqueous solution, and dissolving with a proper amount of solvent to obtain uniform needle fluid; coating a proper amount of needle body liquid on the female die prepared in the step 1), putting the female die into a vacuum drying box, and vacuumizing to enable the needle body liquid to enter holes of the female die; taking out the female die, and scraping the solution outside the holes; and drying in a drying oven for 2 h;

(3) second vacuum mold

Dissolving a backing material into uniform backing liquid by using a proper amount of solvent, coating the backing liquid on the female die prepared in the step (2), and putting the female die into a vacuum drying oven for vacuumizing; and drying in an oven for 4h, and demolding to obtain the product.

Technical Field

The invention relates to the technical field of microneedle administration of vaccines, and provides a thermostable hepatitis A vaccine soluble microneedle patch and a preparation method thereof.

Background

Viral hepatitis type A, referred to as hepatitis A for short, is an infectious disease caused by hepatitis A virus, mainly caused by liver inflammation, mainly transmitted through a feces-oral route, and can be suffered at any age, but mainly used for children and teenagers, and the current measure for effectively preventing the disease is to use hepatitis A vaccine. The existing hepatitis A vaccine is generally administrated by needle injection, and the compliance of patients is poor, so a novel vaccine administration route is urgently needed. Compared with the injection route, oral administration still has the limitation, especially for the biomacromolecule drugs, the bioavailability is reduced due to the influence of the protease of the gastrointestinal tract and the first pass effect of the liver. The field of transdermal administration is currently the most popular field, in addition to the injection and oral routes, and the skin acts as the largest immune organ in the human body, which makes the combination of a vaccine with the field of transdermal administration highly attractive.

A key factor that hinders transdermal delivery of biomacromolecule drugs is the barrier of the stratum corneum, the outermost layer of the skin. The micro-needle is a new physical penetration-promoting technology which develops rapidly in nearly ten years, can break the stratum corneum barrier of the skin to enter the epidermis layer of the skin without touching the nerve endings of the dermis layer, achieves painless minimally invasive administration, and breaks through the administration effect of the traditional transdermal administration preparation. According to different structural and functional characteristics, the existing microneedles can be divided into the following four types, namely solid microneedles, hollow microneedles, coated microneedles and soluble microneedles. Solid microneedles and hollow microneedles are simpler to prepare but can cause harmful residues in the skin if the solid microneedles and the hollow microneedles are accidentally broken; the drug loading of the coated microneedle is small and the dosage is difficult to control; in contrast, soluble microneedles have the advantages of high drug loading, self-dissolution when inserted into the skin, no harmful residue, and the like, and are widely studied.

Korean patent publication No. KR20170032810A provides a technical solution for loading hepatitis a vaccine into soluble microneedles, which, although some water-soluble polymers are used to prepare microneedles and characterization of drug release behavior is performed, simply lists examples of which materials can be used to prepare soluble microneedles, and does not screen specific prescriptions and ratios for solving some technical problems of soluble microneedle administration of hepatitis a vaccine; moreover, the preparation method of the microneedle is a more conventional integrated injection molding method.

In addition, hepatitis b vaccines, similar to hepatitis a vaccines, have been reported in a number of documents loaded with dissolvable microneedles. Such as: the literature "nanoparticles immobilized in dispersed microzyme immobilized and thermostabile" (poison D, Renaud F, Dewar V, et al. biomaterials,2017,145:256-265), the literature "DNA-based catalysis against nanoparticles immobilized in microscopic microparticles attached by cationic ligands and CpG ODN" (Qia Y, Guo L, Zhang S, drug delivery,2016,23(7):2391-2398), and the like. It is known that when the active ingredients of the hepatitis B vaccine are changed compared with the hepatitis A vaccine, different matrix materials are combined with the vaccine drugs to cause the mechanical properties and the effect difference of the soluble microneedles of the vaccine, which is consistent with the conclusion of the literature "matrix materials and composite materials for constructing the soluble microneedles" (octogen, Mafengsen, Haohui et al, material guidance, 2017,31(19): 129-134.). Therefore, if the soluble microneedle prescription of the hepatitis B vaccine is used for developing the soluble microneedle of the hepatitis A vaccine, a plurality of uncertainties still exist.

The hepatitis A vaccine is protein in nature, and is temperature sensitive like a common vaccine. In order to protect the active ingredients of a vaccine against denaturation, it is often necessary to add suitable stabilizers to maintain the activity of the vaccine during storage or drying. According to the literature (Sun Dongpo, Huyi bridge. protein freeze-drying protective agent and its protective mechanism [ J ] pharmaceutical progress, 2003,27(4): 201-; any single protectant may not possess all of the protective properties, and thus the effect of a combination of protectants is often greater than the effect of a single protectant; the protective agent is usually used at an optimal concentration for the protective effect of a vaccine and has an effect on the penetration performance of the microneedles, so that reasonable amounts should be considered when screening for protective agents.

In conclusion, how to develop a soluble microneedle patch for hepatitis a vaccine with appropriate prescription, good thermal stability and moderate mechanical strength is a technical problem which needs to be solved urgently by the technical personnel in the field.

Disclosure of Invention

The invention aims to provide a thermostable hepatitis A vaccine soluble microneedle patch, which solves the bottleneck existing in the prior art.

The technical scheme adopted by the invention is as follows:

a heat-stable hepatitis A vaccine soluble micro-needle patch comprises a needle body and a back lining, wherein the needle body is composed of a hepatitis A vaccine, a matrix material and a vaccine stabilizer, the vaccine stabilizer is a mixture of polyvinylpyrrolidone and dextran, and the content ratio of the vaccine stabilizer to the hepatitis A vaccine is (2:1) - (5: 1).

Preferably, the content ratio of the polyvinylpyrrolidone to the dextran in the vaccine stabilizer is (3:1) - (1: 3).

Preferably, the matrix material is polyvinyl alcohol.

Preferably, the content ratio of the polyvinylpyrrolidone to the dextran in the vaccine stabilizer is 2: 1.

preferably, the needle body comprises the following components in percentage by weight: 30 wt% of polyvinyl alcohol, 37.3 wt% of polyvinylpyrrolidone, 18.7 wt% of dextran and 14 wt% of hepatitis A vaccine.

The invention also provides a preparation method of the heat-stable hepatitis A vaccine soluble microneedle patch, which comprises the following steps:

(1) negative mold preparation of microneedles

Firstly, preparing male mold microneedles by adopting photoetching and etching processes, then mixing polysiloxane and a curing agent according to the mass ratio of 6:1, and pouring the mixture into a cuboid container in which the monocrystalline silicon male mold microneedles are placed; placing the container in a vacuum drying box, and vacuumizing to remove bubbles in the mixed solution; then putting the molded polysiloxane into an oven, drying for 3 hours, and taking out to obtain a molded polysiloxane mold;

(2) first vacuum mold

Mixing the hepatitis A vaccine, the matrix material and the vaccine stabilizer according to the prescription amount in proportion, adding an eosin aqueous solution, and dissolving the eosin aqueous solution into a uniform needle fluid by using a proper amount of solvent; coating a proper amount of needle body liquid on the female die prepared in the step 1), putting the female die into a vacuum drying box, and vacuumizing to enable the needle body liquid to enter holes of the female die; taking out the female die, and scraping the solution outside the holes; and drying in a drying oven for 2 h;

(3) second vacuum mold

Dissolving a backing material into uniform backing liquid by using a proper amount of solvent, coating the backing liquid on the female die prepared in the step (2), and putting the female die into a vacuum drying oven for vacuumizing; and drying in an oven for 4h, and demolding to obtain the product.

Compared with the prior art, the heat-stable hepatitis A vaccine soluble microneedle patch and the preparation method thereof provided by the invention have the following beneficial effects:

① the present invention selects a specific component to content ratio of vaccine stabilizer (PVP and dextran) to be combined with hepatitis A vaccine so that the vaccine in the soluble microneedles can remain stable at high temperatures.

② the matrix material (polyvinyl alcohol) with a specific content ratio is selected to be combined with the hepatitis A vaccine and the vaccine stabilizer, so that the soluble micro-needle has moderate mechanical strength and is effective to penetrate into the skin.

In a word, the soluble microneedle patch for the hepatitis A vaccine prepared by the invention has good heat stability of the needle body medicament and moderate mechanical strength, and can effectively penetrate into the skin.

Drawings

FIG. 1 is a schematic flow chart of a reverse molding method for preparing a microneedle negative mold;

fig. 2 is a schematic flow chart of the preparation of hepatitis a vaccine microneedles according to the present invention.

Detailed Description

In order to make the technical solution of the present invention better understood by those skilled in the art, the technical solution in the present embodiment will be specifically described below with reference to the accompanying drawings in the present application. It should be noted that the following examples are only for illustrating the present invention and are not to be construed as limiting the present invention, and any modifications and changes made to the present invention within the spirit and scope of the claims are included in the scope of the present invention.