阅读说明：本技术 一种乙脑疫苗可溶性微针贴及其制备方法 (Japanese encephalitis 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：本发明提供了一种乙脑疫苗可溶性微针贴,包括针体和背衬,所述的针体由乙脑疫苗、基质材料、佐剂组成,其中,佐剂为MDP与GM-CSF的混合,所述的佐剂与乙脑疫苗的含量比为(1：1)～(1：3)。本发明制备的乙脑疫苗可溶性微针,免疫效果好,针体机械强度高,有望取代乙脑疫苗注射剂,实现无痛微创给药。(The invention provides a Japanese encephalitis vaccine soluble microneedle patch which comprises a needle body and a back lining, wherein the needle body consists of a Japanese encephalitis vaccine, a matrix material and an adjuvant, the adjuvant is a mixture of MDP and GM-CSF, and the content ratio of the adjuvant to the Japanese encephalitis vaccine is (1:1) - (1: 3). The Japanese encephalitis vaccine soluble microneedle prepared by the invention has good immune effect and high mechanical strength of the needle body, is expected to replace Japanese encephalitis vaccine injection, and realizes painless minimally invasive administration.)

1. The soluble micro-needle patch for the Japanese encephalitis vaccine comprises a needle body and a back lining, and is characterized in that the needle body consists of the Japanese encephalitis vaccine, a matrix material and an adjuvant, wherein the adjuvant is a mixture of MDP and GM-CSF, and the content ratio of the adjuvant to the Japanese encephalitis vaccine is (1:1) - (1: 3).

2. The Japanese encephalitis vaccine soluble microneedle patch according to claim 1, wherein the matrix material is hyaluronic acid.

3. The Japanese encephalitis vaccine soluble microneedle patch according to claim 1, wherein the content ratio of GM-CSF to MDP in the adjuvant is (3:1) - (1: 3).

4. The Japanese encephalitis vaccine soluble microneedle patch according to claim 3, wherein the content ratio of GM-CSF to MDP in the adjuvant is 1: 2.

5. the Japanese encephalitis vaccine soluble microneedle patch according to claim 1, wherein the content ratio of adjuvant to Japanese encephalitis vaccine is 1: 2.

6. the Japanese encephalitis vaccine soluble microneedle patch according to claim 1, wherein the matrix material accounts for 30-50% of the needle body by mass.

7. The Japanese encephalitis vaccine soluble microneedle patch according to any one of claims 1 to 6, wherein the needle body contains the following components: the weight percentage of the Japanese encephalitis vaccine is 40 percent, the weight percentage of GM-CSF is 6.7 percent, the weight percentage of MDP is 13.3 percent, and the weight percentage of hyaluronic acid is 40 percent.

8. The method for preparing a Japanese encephalitis vaccine soluble microneedle patch according to claim 1, comprising the steps of:

(1) negative mold preparation of microneedles

Preparing a polydimethoxysiloxane female die required by the Japanese encephalitis vaccine soluble microneedle by adopting a reverse die method;

(2) first-time centrifugal mold-entering

Mixing and dissolving the components according to the prescription amount into uniform needle body liquid, coating a proper amount of the needle body liquid on a female die manufactured by an inverse die method, putting the female die into a 96-hole plate centrifuge, and centrifuging to enable the needle body liquid to enter holes of the female die; then, taking out the female die, and scraping the solution outside the holes;

(3) second time centrifugal mold

Dissolving the backing into uniform backing liquid, coating a proper amount of the backing liquid on the female die obtained in the step (1), putting the female die into a 96-hole plate centrifuge, and centrifuging to enable the backing liquid to enter the holes of the female die; and then putting the mixture into an oven, drying the mixture at 37 ℃, taking out the dried mixture, and demolding to obtain the product.

Technical Field

The invention relates to the technical field of microneedle administration of vaccines, and provides a soluble microneedle patch for a Japanese encephalitis vaccine and a preparation method thereof.

Background

Epidemic encephalitis B is an acute infectious disease affecting the central nervous system caused by encephalitis B virus, often causes death of patients or leaves nervous system sequelae, and encephalitis B vaccine is an effective measure for preventing epidemic encephalitis B. The existing Japanese encephalitis vaccine is generally administered through needle injection, and the compliance of patients is poor, so a novel vaccine administration route is urgently needed. In addition, the protein drugs are easily decomposed by the proteases in the gastrointestinal tract or the first-pass effect of the liver by oral administration, resulting in reduced activity, thereby affecting the bioavailability. Transdermal Drug Delivery System (TDDS) is a new technology of drug delivery which is popular in recent years, which can avoid the problems of poor patient compliance caused by the above-mentioned needle injection and the problems of oral administration of the first pass effect of the digestive tract and liver, and skin is favored as the largest immune organ of human body. However, transdermal delivery of water-soluble small molecule as well as macromolecular drugs has been problematic due to the presence of stratum corneum barriers.

Various physical permeation-promoting techniques are currently used to promote transdermal delivery of drugs, such as microneedles, iontophoresis, electrothermal porogenesis, ultrasound permeation promotion, and magnetic field introduction. Compared with the prior art, other physical permeation-promoting technologies can hardly control the administration position and dosage, and the microneedle can control the administration position by changing the length of the microneedle, and for a soluble microneedle, the microneedle can accurately control the administration dosage by controlling the drug content of the tip of the cast microneedle, so that the microneedle administration has unique advantages in both aspects, and the research in the field of microneedles raises the temperature year by year. The micro needles (microneedles) are an array combination with a plurality of micro needles, can penetrate into the stratum corneum of the skin to enter the epidermis layer without touching the nerve endings of the dermis layer, achieve painless minimally invasive administration, and the administration effect breaks through the traditional transdermal administration preparation. Compared with other types of microneedles, the soluble microneedle has the advantages of large drug loading, capability of being automatically degraded after penetrating into the skin without causing harmful residues, capability of achieving sustained and controlled release by screening different matrix material proportions and the like, and becomes the most studied microneedle type at present.

There is no example of loading Japanese encephalitis vaccine into soluble microneedle in the current domestic and foreign patents and literature. Similarly, patent publication No. IN2008CN02444A discloses that the administration of encephalitis vaccine into coated microneedles is well known to be very different from soluble microneedles IN terms of manufacturing process and drug loading, so it is difficult to use (Indermun S, Luttge R, Choonara Y E, et al. current enhancements IN the manufacture of the fabrics for transdermal delivery [ J ]. Journal of Controlled Release 2014,185(185): 130-.

The adjuvant is a non-specific immunopotentiator which can enhance the immune response of the organism to antigens or change the type of immune response when injected with antigens or injected into the organism in advance, the action mechanism of the immunologic adjuvant can be divided into ① antigen sustained release (antigen repository effect) ② upregulating various cytokines and chemokines, ③ recruiting immune cells to the injection site, ④ enhancing the uptake and presentation of antigens, ⑤ activating Antigen Presenting Cells (APCs), promoting the mature antigen presentation to be transported to draining lymph nodes, ⑥ activating inflammatory bodies, and the like, the immune enhancement effect of different adjuvants or adjuvants is different, therefore, aiming at different vaccines, the influence of different adjuvants and immune pathways on the immune protection effect of Eimeria tenella SO7 antigen can be achieved (J. Proc., 2007, 6929 (9):697, Pengjinpai Bighua, etc.: veterinary No. Shisanjie et al J. Immunity adjuvant, No. Shisanjie et al J. Shisanjie et al., Shisanjie et al.: No. Shi. Shisanjie et al.: 1, No. Shi. Shisanjie et al).

In addition to the advantages mentioned above, dissolvable microneedles suffer from certain limitations, such as possibly insufficient mechanical strength relative to other types of microneedles. According to the reference (Park J H, Allen M G, Prausnitz MR. Polymer microelectronics for controlled-release drug delivery [ J ]. pharmaceutical research,2006,23(5): 1008-. Therefore, screening of suitable microneedle drug loading ratio has a certain meaning for improving the mechanical strength of soluble microneedles.

In summary, how to develop a soluble microneedle with a suitable prescription, a good immune effect and a good mechanical strength for a Japanese encephalitis vaccine is a technical problem which needs to be solved urgently by those skilled in the art.

Disclosure of Invention

The invention aims to provide a Japanese encephalitis vaccine soluble microneedle patch and a preparation method thereof, and solves the bottleneck existing in the prior art.

The technical scheme adopted by the invention is as follows:

the needle body consists of a Japanese encephalitis vaccine, a matrix material and an adjuvant, wherein the adjuvant is a mixture of Muramyl Dipeptide (MDP) and human granulocyte-macrophage colony stimulating factor (GM-CSF), and the content ratio of the adjuvant to the Japanese encephalitis vaccine is (1:1) - (1: 3).

Preferably, the matrix material is Hyaluronic Acid (HA).

Preferably, the content ratio of GM-CSF to MDP in the adjuvant is (3:1) - (1: 3).

More preferably, the content ratio of GM-CSF to MDP in the adjuvant is 1: 2.

preferably, the content ratio of the adjuvant to the Japanese encephalitis vaccine is 1: 2.

preferably, the matrix material accounts for 30-50% of the needle body by mass.

Preferably, the needle body comprises the following components in percentage by weight: the weight percentage of the Japanese encephalitis vaccine is 40%, the weight percentage of GM-CSF is 6.7%, the weight percentage of MDP is 13.3%, and the weight percentage of HA is 40%.

The preparation method of the Japanese encephalitis vaccine soluble microneedle patch comprises the following steps:

(1) negative mold preparation of microneedles

Preparing a polydimethoxysiloxane female die required by the Japanese encephalitis vaccine soluble microneedle by adopting a reverse die method;

(2) first-time centrifugal mold-entering

Mixing and dissolving the components according to the prescription amount into uniform needle body liquid, coating a proper amount of the needle body liquid on a female die manufactured by an inverse die method, putting the female die into a 96-hole plate centrifuge, and centrifuging to enable the needle body liquid to enter holes of the female die; then, taking out the female die and scraping off the solution outside the hole;

(3) second time centrifugal mold

Dissolving the backing into uniform backing liquid, coating a proper amount of the backing liquid on the female die obtained in the step (1), putting the female die into a 96-hole plate centrifuge, and centrifuging to enable the backing liquid to enter the holes of the female die; and then putting the mixture into an oven, drying the mixture at 37 ℃, taking out the dried mixture, and demolding to obtain the product.

Compared with the prior art, the soluble micro-needle patch for the Japanese encephalitis vaccine and the preparation method thereof provided by the invention have the following beneficial effects:

① the invention selects the compound immunologic adjuvant with specific content ratio to combine with the Japanese encephalitis vaccine, so that the Japanese encephalitis vaccine soluble microneedle has the best immune enhancement effect.

② the invention screens proper matrix material mass ratio, so that the Japanese encephalitis vaccine soluble microneedle has moderate mechanical property, and the drug delivery success rate of the microneedle is improved.

③ the soluble microneedle prepared by the invention is a layered microneedle, which can make the dosage of the microneedle easier to control and can not cause drug waste.

In a word, the Japanese encephalitis vaccine soluble microneedle prepared by the invention has good immune effect and high mechanical strength of the needle body, is expected to replace Japanese encephalitis vaccine injection, and realizes painless minimally invasive administration.

Drawings

Fig. 1 is a schematic flow chart of preparing a microneedle negative mold;

FIG. 2 is a schematic flow chart of the process for preparing the Japanese encephalitis 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.