阅读说明：本技术 一种载螨变应原微球的缓释可溶性微针递送系统 (Slow-release soluble microneedle delivery system of mite-loaded allergen microsphere ) 是由 不公告发明人 于 2020-04-30 设计创作，主要内容包括：本发明公开了一种载螨变应原微球的缓释可溶性微针递送系统,包括针体和背衬,针体包括螨变应原微球和基质材料,其中,基质材料为甲基乙烯基醚-顺丁烯酸酐共聚物和聚乙二醇的混合,甲基乙烯基醚-顺丁烯酸酐共聚物和聚乙二醇的含量比为(2:1)～(1:1)。本发明通过选择特定的螨变应原微球的组分种类及含量比,使制得的螨变应原缓释微球性能卓越,能较好的负载螨变应原；并通过选择特定的基质材料组分种类及含量比,及特定的药载比,使制得的螨变应原缓释可溶性微针成型性、机械强度等性能俱佳,且大大延长了药物的释放时间,治疗效果优于非缓释型的螨变应原可溶性微针,有利于降低螨变应原给药频次,增加患者的依从性。(The invention discloses a slow-release soluble microneedle delivery system carrying mite allergen microspheres, which comprises a needle body and a back lining, wherein the needle body comprises mite allergen microspheres and a matrix material, the matrix material is a mixture of methyl vinyl ether-maleic anhydride copolymer and polyethylene glycol, and the content ratio of the methyl vinyl ether-maleic anhydride copolymer to the polyethylene glycol is (2:1) - (1: 1). According to the invention, by selecting the component types and content ratios of the specific mite allergen microspheres, the prepared mite allergen slow-release microspheres have excellent performance and can better load mite allergens; and by selecting the specific types and content ratios of the components of the matrix material and the specific drug loading ratio, the prepared mite allergen sustained-release soluble microneedle has good moldability, mechanical strength and other properties, greatly prolongs the release time of the drug, has better treatment effect than non-sustained-release mite allergen soluble microneedles, is beneficial to reducing the administration frequency of the mite allergen and increases the compliance of patients.)

1. A slow-release soluble microneedle delivery system carrying mite allergen microspheres comprises a needle body and a back lining, and is characterized in that the needle body comprises mite allergen microspheres and a matrix material, wherein the matrix material is a mixture of a methyl vinyl ether-maleic anhydride copolymer and polyethylene glycol, and the content ratio of the methyl vinyl ether-maleic anhydride copolymer to the polyethylene glycol is (2:1) - (1: 1).

2. The mite allergen microsphere-loaded sustained-release soluble microneedle delivery system according to claim 1, wherein the content ratio of the methyl vinyl ether-maleic anhydride copolymer to the polyethylene glycol is 3: 2.

3. The mite allergen microsphere-loaded sustained-release soluble microneedle delivery system according to claim 1, wherein the content ratio of the mite allergen microspheres to the matrix material is (2:3) - (1: 1).

4. The mite allergen microsphere-loaded sustained-release soluble microneedle delivery system according to claim 3, wherein the content ratio of the mite allergen microspheres to the matrix material is 1: 1.

5. The mite allergen microsphere-loaded slow-release soluble microneedle delivery system according to claim 1, wherein the mite allergen microsphere is composed of an oil phase, an inner water phase and an outer water phase, wherein the oil phase is a blended solution formed by dissolving PEG-PLGA and micro span 60 in dichloromethane, the inner water phase is a mite allergen aqueous solution, the outer water phase is a 0.5% PVA aqueous solution, and the volume ratio of the oil phase to the inner water phase is 14: 1.

6. The mite allergen microsphere-loaded slow-release soluble microneedle delivery system according to any one of claims 1 to 5, wherein the needle body comprises the following components in percentage by weight: the mass ratio of the mite allergen microsphere is 50%, the mass ratio of the methyl vinyl ether-maleic anhydride copolymer is 30%, and the mass ratio of the polyethylene glycol is 20%.

Technical Field

The invention relates to the technical field of administration of mite allergen, in particular to a mite allergen microsphere-loaded slow-release soluble microneedle delivery system.

Background

Mites are important allergen sources for inducing allergic diseases such as asthma, allergic rhinitis and eczema. In recent years, research and research studies in a plurality of countries such as the United states, New Zealand and Australia successively prove that the close relationship between mite-derived allergens and allergic diseases is the cause of the allergic diseases such as allergic asthma; desensitization therapy is becoming the most effective treatment for this type of disease. However, the mite allergen is mostly administered in the form of injection, and long-term injection causes poor patient compliance, so a new dosage form for delivery of the mite allergen is urgently needed.

The skin is the largest organ of the human body and has a large number of antigen presenting cells, e.g., langerhans cells in the epidermis, accounting for 3% to 5% of the total number of all cells. However, the stratum corneum barrier of the skin makes the delivery of macromolecular drugs such as mite allergens problematic. As a new formulation proposed in 1976, a transdermal drug delivery technology of microneedles has been rapidly developed in the last decade and is favored by researchers. The transdermal drug delivery system can puncture stratum corneum of human skin to enter the epidermis layer without touching nerve endings of the dermis layer, thereby realizing painless minimally invasive drug delivery, and the drug delivery effect breaks through the traditional transdermal drug delivery preparation. The soluble microneedle belongs to one type of microneedles, and has the advantages of large drug loading capacity, controllable drug administration dose and the like compared with other types of microneedles, so that the soluble microneedle is the most widely researched microneedle type at present.

Microspheres are a dispersion of microparticles of a drug dispersed or adsorbed in a polymeric, polymeric matrix. The sustained and controlled release of the medicine can be realized through the micro-encapsulation, and dozens of medicines prepared by the micro-encapsulation technology in recent years are the mature new preparation technology at present. Allergic disease treatments typically employ long-term desensitization therapy, i.e., the body is made tolerant to a particular allergen by frequent administration over a long period of time. Therefore, the development of sustained release formulations of mite allergens can reduce the number of administrations of allergens and increase patient compliance.

By consulting domestic and foreign documents and patents, the mite allergen is directly loaded into the soluble microneedle in the documents and the patents: preparing soluble microneedle delivery mite allergens with hyaluronic acid as in Kim et al (Kim J H, Shin J U, Kim S H, et al. succinsful transmural allergen delivery and allergen-specific immunological use biochemical patches [ J ] Biomaterials,2018,150: 38-48); japanese patent publication No. JP2014156433A and Chinese patent publication No. CN109715075A teach the selection of materials when soluble microneedles are directly loaded with allergens, but do not relate to mite allergen-sustained release microneedles.

In addition, there is no delivery of mite allergen microspheres, and although there is delivery of pollen allergen and bee venom allergen microspheres (adaptive Allergy Vaccine Based on CpG-and Protamine-conjugation PLGA Microparticles [ J ]. Pharmaceutical Research,2007,24(10):1927 and 1935.), there is uncertainty about how to have a good loading effect on mite allergen because of the component difference between the protein component of mite allergen Vaccine and pollen allergen and bee venom allergen.

In addition, although researchers have prepared microsphere-loaded soluble microneedles, none have achieved superior mechanical properties by corresponding prescription screening (Ke C J, Lin Y J, Hu Y C, et al. multidrug release base on microneedles array filters with pH-responsive PLGA hold microspheres [ J ] Biomaterials,2012,33(20):5156-, 2017,31(19): 129-134)) that the combination of different materials and the combination of the materials and the drug can cause the mechanical property of the soluble microneedle to have difference; according to the literature (Park J H, Allen M G, Prausnitz M R. Polymer semiconductors for controlled-release drug delivery [ J ]. Pharmaceutical Research,2006,23(5):1008-19), as the amount of drug in the microneedle formulation increases, the mechanical properties of the microneedles will decrease. Therefore, what kind of material is selected to develop the slow-release soluble microneedle delivery system carrying the mite allergen microspheres and what mass ratio of each component in the soluble microneedle is, so that the soluble microneedle delivery system has a good and effective delivery effect and is still full of uncertainty.

In summary, how to develop a sustained-release soluble microneedle delivery system with excellent mechanical properties and good and effective sustained-release delivery effect for mite allergen vaccine is a technical problem that needs to be solved urgently by those skilled in the art.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a mite allergen-carrying sustained-release soluble microneedle delivery system (short for mite allergen sustained-release soluble microneedle, the same below), and the defects of the prior art are overcome by selecting the proper types and content of the matrix materials and the component types and content ratio of the mite allergen microspheres.

Therefore, the invention adopts the following technical scheme:

a slow-release soluble microneedle delivery system carrying mite allergen microspheres comprises a needle body and a back lining, wherein the needle body comprises the mite allergen microspheres and a matrix material, the matrix material is a mixture of methyl vinyl ether-maleic anhydride copolymer and polyethylene glycol, and the content ratio of the methyl vinyl ether-maleic anhydride copolymer to the polyethylene glycol is (2:1) - (1: 1).

Preferably, the content ratio of the methyl vinyl ether-maleic anhydride copolymer to the polyethylene glycol is 3: 2.

Preferably, the content ratio of the mite allergen microspheres to the matrix material is (2:3) - (1: 1).

More preferably, the content ratio of the mite allergen microspheres to the matrix material is 1: 1.

Preferably, the mite allergen microsphere consists of an oil phase, an inner water phase and an outer water phase, wherein the oil phase is a blended solution formed by dissolving PEG-PLGA and trace span 60 in dichloromethane, the inner water phase is a mite allergen aqueous solution, the outer water phase is a PVA aqueous solution with the concentration of 0.5%, and the volume ratio of the oil phase to the inner water phase is 14: 1.

Preferably, the needle body comprises the following components in percentage by weight: the mass ratio of the mite allergen microsphere is 50%, the mass ratio of the methyl vinyl ether-maleic anhydride copolymer is 30%, and the mass ratio of the polyethylene glycol is 20%.

In the present invention, the type of the methyl vinyl ether-maleic anhydride copolymer (Gantrez) is not limited as long as the mechanical properties of the soluble microneedle prepared therefrom are satisfactory, including but not limited to Gantrez S-97.

Compared with the prior art, the slow-release soluble microneedle delivery system of the mite-loaded allergen microsphere has the following beneficial effects:

according to the invention, by selecting the component types and content ratios of the specific mite allergen microspheres, the prepared mite allergen slow-release microspheres have excellent performance and can better load mite allergens; and by selecting the specific types and content ratios of the components of the matrix material and the specific drug loading ratio, the prepared mite allergen sustained-release soluble microneedle has good moldability, mechanical strength and other properties, greatly prolongs the release time of the drug, has better treatment effect than non-sustained-release mite allergen soluble microneedles, is beneficial to reducing the administration frequency of the mite allergen and increases the compliance of patients.

Drawings

Fig. 1 is a release curve diagram of mite allergen sustained-release soluble microneedles.

Figure 2 is a graph of the release profile of mite allergen soluble microneedles.

Detailed Description

The following are specific examples of the present invention and further describe the technical solutions of the present invention, but the scope of the present invention is not limited to these examples. All changes, modifications and equivalents that do not depart from the spirit of the invention are intended to be included within the scope thereof.

Example 1 preparation of mite allergen sustained-release soluble microneedle

The mite allergen slow-release soluble microneedle to be prepared by the invention consists of mite allergen microspheres and a microneedle substrate material.

Among the preparation methods of the microspheres, the multiple emulsion solvent evaporation method is commonly used for encapsulating the water-soluble core material due to the advantages of simple operation, convenient control of process parameters, no need of pH adjustment and large temperature change, greatly improved drug-loading rate and encapsulation rate of the microspheres compared with a single emulsion system, controlled release and the like. Therefore, the mite allergen microsphere is prepared by adopting a multiple emulsion solvent evaporation method, and the specific method comprises the following steps:

dissolving the capsule wall material and trace span 60 in dichloromethane to serve as an oil phase, taking a mite allergen aqueous solution as an internal water phase, mixing the internal water phase and the oil phase according to a certain proportion, and performing ultrasonic emulsification to obtain the W/O colostrum. And then, injecting the primary emulsion into an external water phase of a PVA (polyvinyl alcohol) aqueous solution with the concentration of 0.5%, and stirring for 20-30 s by a homogenizer to obtain the multiple emulsion. Stirring to volatilize for 3h, removing organic solvent, centrifuging for 15min, discarding supernatant, washing with distilled water, and repeating for 3 times. Freeze-drying to obtain dry powder, and storing at-4 deg.C for use.

The invention adopts a repeated filling mold method to prepare the mite allergen sustained-release soluble microneedle, and the steps are as follows:

1) mixing the mite allergen microsphere and the microneedle matrix material in the formula in proportion, dissolving the mixture into a uniform pouring solution by using a proper amount of solvent (deionized water), filling the pouring solution into a centrifuge tube, placing the centrifuge tube into a centrifugal precipitator, centrifuging to remove bubbles in the pouring solution, and standing for later use.

2) Pouring the pouring solution obtained after centrifugation in the step 1) on a Polydimethylsiloxane (PDMS) female mold, and repeatedly extruding the pouring solution into a PDMS female mold hole by using a metal male mold microneedle;

3) and (3) observing the PDMS female mold, filling the holes of the PDMS female mold with the pouring solution, taking out the mold, putting the mold into an oven, drying the mold at 37 ℃ for 6 hours, taking out the mold, and demolding to obtain the finished product.

Example 2 related assay of mite allergen sustained Release soluble microneedles

1. Particle size of mite allergen microsphere

Precisely weighing the prepared dry microspheres, uniformly spreading the dry microspheres on a glass slide, observing the shape by using an optical microscope with a micrometer, counting the particle sizes (mum) of 300 microspheres, dividing the particle size range, and calculating the average particle size by statistical treatment.

When the average particle size of the microspheres is less than 30 micrometers, the microspheres are more easily distributed on the tips of the microneedles, are considered to meet the requirements, and can be used for subsequent preparation.

2. Determination of encapsulation efficiency of mite allergen microsphere

The total protein content of 10ml of the mite allergen vaccine solution was determined by the Coomassie Brilliant Blue (CBB) method and was designated W1. And then redissolving the prepared mite allergen microspheres, carrying out vortex oscillation for 5min, then ultracentrifuging for 16000rpm and 10min, taking supernate, measuring the total protein content by using a CBB method, marking as W2, and calculating the encapsulation efficiency EE. The encapsulation efficiency EE is (W1-W2)/W1 is 100%.

According to the requirements of 'Chinese pharmacopoeia' (general rule 9014) of 2015 edition, when the encapsulation rate of the microspheres is more than 80%, the microspheres meet the requirements.

3. Mechanical property determination of mite allergen sustained-release soluble microneedle

1) The moldability of the microneedles was judged by observing the needle content (soluble microneedle number/positive mold number 100%, parallel 6 groups) of the microneedles under a scanning electron microscope SEM, and the higher the needle content, the better the moldability of the microneedles.

2) Placing a weight of 20N on the microneedle with the upward needle point, taking off the weight after 3min, observing the residual height percentage of the microneedle (the residual height percentage is the height of the soluble microneedle/the height of the male mold microneedle 100%, and paralleling 6 groups) by using a microscope to judge the mechanical strength of the microneedle, wherein the larger the residual height percentage is, the better the mechanical strength of the microneedle is.

When the needle content of the micro-needle is more than 80 percent and the residual height percentage is more than 70 percent, the micro-needle is considered to meet the test requirement.

4. Drug release performance investigation of mite allergen

PBS buffer solution with pH 7.4 is prepared, and a Franz diffusion cell method is adopted: taking 6 mite allergen sustained-release soluble microneedle patches prepared by confirming a prescription, and loading the microneedle on a Franz diffusion cell after penetrating a paraffin parafilm sealing membrane in advance. Samples were taken at 1d, 2d, 3d, 4d, 5d, 6d, 7d, 8d, and 9d, respectively, at a sample volume of 0.9 mL. The receiving solution was PBS buffer with pH 7.4.

The Franz diffusion method was also used: taking 6 mite allergen soluble microneedle patches with known drug loading capacity, and loading the microneedles on a Franz diffusion cell after puncturing a parafilm sealing film in advance. Sampling is carried out for 30s, 1min, 5min, 10min, 30min, 1h, 3h, 6h and 10h respectively, and the sampling amount is 0.9 mL. The receiving solution was PBS buffer with pH 7.4.

And finally, detecting the content of the mite allergen in the sampled solution in different periods by an ELISA kit, and drawing a drug release curve.

5. Determination of therapeutic Effect of mite allergen

The prepared mite allergen soluble microneedle and 9 pieces of mite allergen slow-release soluble microneedle are respectively taken, 18 sensitized rat back skins after depilation are respectively punctured by using an administration device, and the microneedle parts are taken out after being completely dissolved. Serum was taken from rat veins at intervals of 0 day, 7 days, and 14 days after administration, respectively, and the level of specific IgE in the serum was quantified by ELISA. The initial serum-specific IgE levels of the allergic rats were measured on day 0, and the serum-specific IgE levels measured on days 7 and 14 are indicative of the therapeutic effect on the allergic rats after administration, and the lower the serum-specific IgE level, the better the therapeutic effect.

Example 3 influence of component types and content of mite allergen microsphere on mite allergen Loading

In this embodiment, the mite allergen microsphere comprises a three-phase system composed of an oil phase, an inner water phase and an outer water phase, the oil phase is a blended solution formed by dissolving a capsule wall material and micro span 60 in dichloromethane, the inner water phase is a mite allergen aqueous solution, the outer water phase is a polyvinyl alcohol (PVA) aqueous solution with a concentration of 0.5%, and the influence of the mite allergen microsphere constructed by different materials on the encapsulation efficiency and the particle size shown in table 1 is explored by changing the type of the capsule wall material and the volume ratio of the oil phase to the inner water phase.

In this embodiment, the capsule wall material is selected from one of poly (lactic-co-glycolic acid) (PLGA), poly (ethylene glycol) -poly (lactic-co-acrylic acid) (PEG-PLGA), poly (ethylene glycol) -poly (lactic acid) (PEG-PLA), and poly (3-hydroxybutyrate) (PHB); the content ratio of the oil phase to the inner water phase is preliminarily determined according to a pre-experiment, and is selected to be different from 20:1 to 2:1 (table 1.).

In this example, see example 1 for a process for preparing mite allergen microspheres. The encapsulation efficiency and the average particle size of the mite allergen microsphere are determined in example 2, and the test results are shown in table 1.

TABLE 1 influence of mite allergen microspheres constructed from different materials on encapsulation efficiency and particle size (mean + -SD, n ═ 6)

As can be seen from Table 1, the encapsulation efficiency and the particle size of the mite allergen microsphere are comprehensively considered, and the test requirements are met only when the capsule wall material is PEG-PLGA and the content ratio of the oil phase to the internal water phase is 14: 1.

Therefore, in a three-phase system forming the mite allergen microsphere, a blended liquid formed by dissolving PEG-PLGA and trace span 60 in dichloromethane is selected as an oil phase, a mite allergen aqueous solution is selected as an inner water phase, a PVA aqueous solution with the concentration of 0.5% is selected as an outer water phase, and the content ratio of the oil phase to the inner water phase is 14: 1.

Example 4 Effect of the types of Components of the matrix Material on the Properties of mite allergen-sustained Release soluble microneedles

In example 3, the ratio of the matrix material to the needle body was set to a constant value (60% of the matrix material and 40% of the mite allergen microsphere particles), and the influence of different combinations of matrix materials on the mechanical properties of microneedles was examined by changing only the types of the components of the matrix material as shown in table 2.

In this embodiment, the matrix material is selected from one or a combination of two of methyl vinyl ether-maleic anhydride copolymer (Gantrez), polyethylene glycol (PEG), Hyaluronic Acid (HA) and maltose.

In this example, the process for preparing mite allergen sustained-release soluble microneedles is described in example 1. The method for measuring the formability and mechanical strength of the microneedle is shown in example 2, and the test results are shown in Table 2.

TABLE 2 Effect of different matrix Material compositions on microneedle Performance (mean SD)

As can be seen from Table 2, the pin ratio and the remaining height percentage were considered in combination, and the pin ratio was 95.6. + -. 1.4% and the remaining height percentage was 72.5. + -. 2.5% only when the matrix material was a mixture of Gantrez and PEG (i.e., group No. 7), which all met the test requirements.

Therefore, a mixture of Gantrez and PEG was chosen as the matrix material.

Example 5 influence of content ratio of each component between matrix materials on mite allergen sustained-release soluble microneedle performance

On the basis of example 4, the mass ratio of the matrix material in the needle body was set to a constant value (60% of the matrix material and 40% of the mite allergen microsphere particles), and the components of the matrix material were fixed as a mixture of Gantrez and PEG, and the influence of the matrix material components with different content ratios on the microneedle performance was examined by changing only the content ratio of Gantrez to PEG as shown in table 3.

In this example, the process for preparing mite allergen sustained-release soluble microneedles is described in example 1. The method for measuring the formability and mechanical strength of the microneedle is shown in example 2, and the test results are shown in Table 3.

TABLE 3 influence of different content ratios of matrix material components on microneedle Performance (mean + -SD)

No.	Content ratio of Gantrez to PEG	Needle content (%)	Percentage of remaining height (%)
				1	Gantrez:PEG＝3:1	94.2±1.4	69.5±1.7
2	Gantrez:PEG＝2:1	95.6±1.7	71.1±2.1
				3	Gantrez:PEG＝3:2	98.3±1.6	83.2±2.3
4	Gantrez:PEG＝1:1	95.8±1.4	72.5±2.5
				5	Gantrez:PEG＝2:3	93.1±1.2	64.1±1.4

As can be seen from Table 3, the test requirements were satisfied when the content ratio of Gantrez to PEG was (2:1) to (1:1) in consideration of the needle content and the percentage of remaining height. Among them, when the content ratio of Gantrez to PEG was 3:2 (i.e., No.3 group), the remaining height percentage and the needle-containing rate were the highest, indicating that the mechanical properties and moldability of the microneedles were the best.

Therefore, 3:2 was chosen as the optimal content ratio of Gantrez to PEG.

Example 6 Effect of drug Loading ratio on mite allergen sustained Release soluble microneedle Performance

According to the literature (Park J H, Allen M G, Prausnitz M r. polymer semiconductors for controlled-release drug delivery [ J ]. Pharm Res,2006,23(5):1008-19.), variations in the amount of drug loaded in the microneedles affect the mechanical properties of the microneedles and are uncertain.

Based on example 5, the composition and ratio of the matrix material were set to a constant value, and the effect of different drug loading ratios on the microneedle performance as shown in table 4 was examined by changing only the drug loading ratio (i.e., the content ratio of the mite allergen microspheres to the matrix material).

In this example, the process for preparing mite allergen sustained-release soluble microneedles is described in example 1. The method for measuring the formability and mechanical strength of the microneedle is shown in example 2, and the test results are shown in Table 4.

TABLE 4 microneedle formulations and effects on microneedle Performance (mean + -SD) with different drug loading ratios

No.	Drug loading ratio	Needle content (%)	Percentage of remaining height (%)
				1	2:3	98.3±1.6	83.2±2.3
2	3:4	97.8±1.4	77.2±2.1
				3	1:1	97.6±1.6	75.3±2.4
4	4:3	83.6±1.2	67.2±1.8
				5	3:2	75.6±2.2	54.1±3.1

As can be seen from Table 4:

1) the needle content and the residual height percentage are comprehensively considered, and the test requirements are met when the drug loading ratio is (2:3) - (1: 1).

2) Considering the influence of drug loading on the treatment effect of the mite allergen, the optimal prescription is selected when the drug loading ratio is 1: 1.

In summary, in the mite allergen sustained-release soluble microneedle provided by the invention, the optimal prescription of the needle body is as follows: the mass ratio of the mite allergen microspheres is 50%, the mass ratio of the Gantrez is 30%, and the mass ratio of the PEG is 20%; wherein, the three-phase system of the mite allergen microsphere comprises: the oil phase is a blended solution formed by dissolving PEG-PLGA and trace span 60 in dichloromethane, the inner water phase is a mite allergen aqueous solution, the outer water phase is a PVA aqueous solution with the concentration of 0.5%, and the content ratio of the oil phase to the inner water phase is 14: 1.

In the mite allergen sustained-release soluble microneedle provided by the invention, a proper prescription of the needle body is as follows: the content ratio of the mite allergen microspheres to the matrix material is (2:3) - (1:1), the matrix material consists of Gantrez and PEG, and the content ratio of Gantrez to PEG is (2:1) - (1: 1).

Example 7 examination of Release Curve of mite allergen-releasing soluble microneedles

In the embodiment, the release curves of the mite allergen slow-release soluble microneedle and the mite allergen soluble microneedle are compared, only the mite allergen microsphere and other doses are replaced by the mite allergen soluble microneedle, the mite allergen soluble microneedle is directly loaded into the microneedle, the matrix material is unchanged, and the rest of preparation methods refer to the embodiment 1, so that the mite allergen soluble microneedle can be prepared.

In this example, the process for preparing mite allergen sustained-release soluble microneedles is described in example 1. The drug release performance of mite allergen was examined and shown in example 2, and the release curve is shown in fig. 1 and fig. 2.

From the results of fig. 1 and 2, it can be seen that: the release time of the mite allergen sustained-release soluble microneedle provided by the invention is 8 days, and the accumulated drug release amount exceeds 90%; while the release time of the common mite allergen soluble micro-needle (not slow release) is 180min, the cumulative drug release amount exceeds 90%.

Example 8 evaluation of therapeutic Effect of mite allergen-releasing soluble microneedle

In this example, the therapeutic effects of the mite allergen slow-release soluble microneedle and the mite allergen soluble microneedle are compared, only the mite allergen microspheres with the same dosage are replaced by the mite allergen, the mite allergen is directly loaded into the microneedle, the matrix material is not changed, and the rest of the preparation methods refer to example 1, so that the mite allergen soluble microneedle can be prepared.

In this example, the process for preparing mite allergen sustained-release soluble microneedles is described in example 1. The therapeutic effect of mite allergen was measured in example 2, and the measurement results are shown in Table 5.

TABLE 5 comparison of therapeutic effects (mean + -SD, n ═ 9)

As can be seen from table 5, the serum IgE level of the mite allergen-soluble microneedle group was lower than that of the mite allergen sustained-release soluble microneedle group when blood was taken on day 7; however, the mite allergen sustained-release soluble microneedle group had a lower serum IgE level than the mite allergen soluble microneedle group at the time of blood drawing on day 14.

The reason for analysis by comparing the release curves of example 7 shows that the drug of the mite allergen slow-release soluble microneedle group is not completely released on day 7, but the drug of the mite allergen slow-release soluble microneedle group is completely released already, so the therapeutic effect of the mite allergen slow-release soluble microneedle group is not as good as that of the mite allergen soluble microneedle group. However, the mite allergen slow-release soluble microneedle group continuously releases the mite allergen with the lapse of time, so that the mite allergen slow-release soluble microneedle group has a better treatment effect on allergic rats at day 14.