Micro-needle patch based on hierarchical targeting mechanism as well as preparation method and application of micro-needle patch

Abstract

The invention relates to the technical field of androgen-derived alopecia, and discloses a micro-needle patch based on a hierarchical targeting mechanism as well as a preparation method and application of the micro-needle patch. The finasteride is arranged on the needle tip section of the microneedle, the quercetin is arranged on the needle root section, different cells are targeted through the finasteride and the quercetin in a graded mode, and the microneedle patch with a graded targeting mechanism is formed. After the micro needle pierces into the skin, the water-soluble polymer at the needle tip section is dissolved, finasteride targeted hair papilla cells are released, and then in the process that finasteride acts, the quercetin slowly released at the needle root section gradually acts on endothelial cells in a targeted mode, the proliferation and migration activity of the endothelial cells is promoted, and the effect of the finasteride on the skin is achieved. Endothelial cells are enhanced to form a highly branched capillary network to act on hair papilla cells, the metabolic demand of the hair papilla cells is maintained, and finally the effect of enhancing the finasteride targeting effect is achieved. Therefore, the microneedle patch with the hierarchical targeting mechanism has an excellent AGA curative effect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of androgenetic alopecia, and particularly relates to a microneedle patch based on a hierarchical targeting mechanism for treating androgenetic alopecia and a preparation method thereof. BACKGROUND

[0002] Androgenetic alopecia (AGA) is the most common type of progressive hair loss, accounting for about 95% of all hair loss types. In this disease, the hair loss rate exceeds the hair growth rate, and the hairline gradually moves up, and the number of hairs decreases.

[0003] In the prior art, there are many drugs and treatment methods for AGA. For example, a Chinese patent with the application publication number CN119587557A discloses the application of D-mannose in preventing and treating androgenetic alopecia, a Chinese patent with the application publication number CN113056269A discloses the use of plasminogen activator inhibitor 1 (PAI-1) inhibitor, and a Chinese patent with the application publication number CN119955665A discloses plant lactobacillus ZJUIDS24 with improved androgenetic alopecia and the application thereof. However, these drugs for AGA are all aimed at finding new substances for treating AGA, without considering how to improve the effect of existing drugs for treating AGA.

[0004] The microneedle for drug loading is generally composed of a microneedle body and a substrate. The needle body is mainly used for drug loading. After penetrating the skin, the microneedle body material is dissolved or biodegraded, so as to achieve accurate drug release. The substrate is connected with the patch. When used, the patch is attached to the skin surface, and the microneedle can penetrate the stratum corneum, the outermost layer of the skin, so as to achieve the dissolution of the microneedle body material and the release of the drug.

[0005] For androgenetic alopecia, finasteride and minoxidil are the existing drugs with certain curative effect. However, the action mechanism of finasteride and minoxidil has significant limitations, and there is an urgent need to break through the existing curative effect bottleneck in clinical treatment and explore safer and more efficient intervention means or treatment schemes. SUMMARY

[0006] In order to solve the technical problem of improving the AGA treatment effect of finasteride, the present application provides a microneedle patch based on a hierarchical targeting mechanism and a preparation method and application thereof.

[0007] The specific technical scheme of the present application is as follows: In a first aspect, the present application provides a microneedle patch based on a hierarchical targeting mechanism, which comprises a microneedle and a patch. The material forming the microneedle body comprises a water-soluble polymer. The microneedle body comprises a needle tip segment and a needle root segment. The needle tip segment is loaded with finasteride, and the needle root segment is loaded with quercetin.

[0008] For AGA, finasteride is the existing drug with certain curative effect, which belongs to 5 alpha reductase inhibitor, mainly acts on hair papilla cells, inhibits the conversion of testosterone (HT) to dihydrotestosterone (DHT), improves the symptoms of hair loss, promotes the growth of hair, and achieves the effect of treating AGA. But finasteride is limited by its mechanism, and its existing curative effect is highly bottlenecked, so it is necessary to explore safer and more efficient intervention means to improve the therapeutic effect of finasteride.

[0009] Therefore, the present application provides a microneedle patch with a hierarchical targeting mechanism, which sets finasteride at the needle tip segment of the microneedle and sets quercetin at the needle root segment. The quercetin set at the needle root segment promotes the targeting of finasteride to hair papilla cells and inhibits the conversion of testosterone to DHT in hair papilla cells, thereby improving the therapeutic effect of finasteride on AGA.

[0010] The present application sets finasteride at the needle tip segment of the microneedle and quercetin at the needle root segment to form a microneedle patch with a hierarchical targeting mechanism. The principle of the hierarchical targeting mechanism to promote curative effect is that after the microneedle is inserted into the skin, the water-soluble polymer at the needle tip segment dissolves to release finasteride targeting hair papilla cells, thereby inhibiting the conversion of testosterone to DHT in hair papilla cells. In the process of finasteride targeting hair papilla cells, quercetin released from the needle root segment gradually targets endothelial cells, promotes the proliferation and migration activity of endothelial cells, thereby strengthening the formation of a highly branched capillary network in endothelial cells to maintain the metabolic demand of hair papilla cells, and ultimately enhances the targeting effect of finasteride. The needle root segment is relatively thicker than the needle tip segment, and it penetrates the skin more shallowly and contacts less liquid, so the drug release rate of the needle root segment is relatively slower than that of the needle tip segment. The relatively slow release of quercetin gradually affects endothelial cells, thereby improving the therapeutic effect of finasteride on AGA.

[0011] As a preferred embodiment of the above-mentioned microneedle patch, the quercetin is loaded by metal organic framework and then loaded by the water-soluble polymer to form the needle root segment.

[0012] By loading the metal organic framework, the quercetin is released more slowly, which can continuously improve the metabolism of hair papilla cells during the process of finasteride targeting hair papilla cells and inhibiting the conversion of testosterone to DHT in hair papilla cells.

[0013] After the microneedle pierces the skin, the tip segment dissolves faster than the root segment because of its sharp tip, so the microneedle releases the quercetin in the root segment relatively slowly, and after being loaded by the metal organic framework, more quercetin can be absorbed by the endothelial cells in the middle and late stages of the action of finasteride. In the middle and late stages of the action of finasteride on the hair papilla cells, the process of converting endogenous testosterone to DHT in the metabolism of hair papilla cells has been inhibited, and promoting the proliferation of endothelial cells and the metabolism of hair papilla cells is more conducive to the inhibition of the conversion of endogenous testosterone to DHT. The enhanced metabolism of hair papilla cells is conducive to the proliferation and differentiation of hair follicle stem cells by secreting Wnt and other signal molecules, thereby improving the symptoms of hair loss and promoting hair growth.

[0014] Further preferably, the metal organic framework is ZIF-8.

[0015] As a preferred microneedle patch, the water-soluble polymer is selected from one or more of hyaluronic acid, sodium alginate, and dextran.

[0016] As a preferred microneedle patch, the finasteride loaded on the tip segment is a liposome.

[0017] In a second aspect, the present application provides a preparation method of a microneedle patch based on a hierarchical targeting mechanism, which comprises the following steps: Step S1, a step of preparing finasteride liposomes to form finasteride liposomes; Step S2, a step of preparing a metal organic framework and loading quercetin to form a quercetin-loaded metal organic framework Qu@MOF; Step S3, mixing the finasteride liposomes with the hyaluronic acid solution to obtain the tip segment material of the microneedle, and mixing the Qu@MOF with the hyaluronic acid solution to obtain the root segment material of the microneedle; Step S4, sequentially pouring the tip segment material and the root segment material into a microneedle mold, drying, and forming a microneedle.

[0018] As a preferred preparation method, the step of preparing finasteride liposomes comprises: Mix 10-15 mg glycerin monostearate and 5-10 mg finasteride, heat to 70-90 DEG C until glycerin monostearate melts, and dissolve 10-30 muL oleic acid in the lipid base to obtain an organic phase; add soybean lecithin and emulsifier to 10-25 mL distilled water, heat to 70-90 DEG C to obtain an aqueous phase, wherein the concentration of the soybean lecithin is 0.5-2 g / mL, and the concentration of the emulsifier is 2-4 g / mL; mix the organic phase and the aqueous phase, stir to form a primary emulsion, and ultrasonically treat to form a fine emulsion; add the fine emulsion to 20-35 mL water containing 2-4 g / mL emulsifier, cool, and obtain a finasteride liposome dispersion.

[0019] Further, the emulsifier is preferably Tween-80.

[0020] As a preferred embodiment of the preparation method, the step of preparing the metal organic framework and loading quercetin comprises: Disperse 2-methylimidazole and quercetin in methanol to obtain a 2-methylimidazole mixture; mix the 2-methylimidazole mixture with a zinc nitrate solution, stir, and collect white solids to obtain the metal organic framework loaded with quercetin.

[0021] As a preferred embodiment of the preparation method, the drying temperature is 30-40 DEG C, and the drying time is 4-8 hours.

[0022] In a third aspect, the application provides a use of a microneedle patch in treatment of androgenetic alopecia.

[0023] Compared with the prior art, the application has the following technical effects: (1) The application provides a microneedle patch with a hierarchical targeting mechanism, which sets finasteride at the needle tip segment of the microneedle and sets quercetin at the needle root segment, and targets different cells through finasteride and quercetin to form a microneedle patch with a hierarchical targeting mechanism. After the microneedle of the application pierces the skin, the water-soluble polymer at the needle tip segment dissolves to release finasteride to target hair papilla cells, and then inhibit the conversion of testosterone into DHT in the hair papilla cells. In the process of targeting the hair papilla cells by finasteride, quercetin released from the needle root segment gradually targets endothelial cells to promote the proliferation and migration activity of the endothelial cells, so that the endothelial cells form a highly branched capillary network to act on the hair papilla cells to maintain the metabolic demand of the hair papilla cells, and finally enhance the targeting effect of finasteride. Therefore, the microneedle patch with a hierarchical targeting mechanism of the application has higher AGA curative effect than a microneedle patch with a single finasteride drug, and also has higher AGA curative effect than a finasteride and quercetin combined application of a smearing type drug.

[0024] (2) The drug delivery system of the drug-loaded microneedle has micron-level size, has many advantages, can directly and non-invasively penetrate the skin by puncture, realizes precise targeting of the drug while avoiding pain, improves the bioavailability of the drug, reduces the frequency of drug administration, and thus improves the compliance of patients. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 is a structural schematic diagram of a microneedle patch based on a hierarchical targeting mechanism; Figure 2 is Figure 1 is a structural schematic diagram of a microneedle in the microneedle patch; Figure 3 is a record photo of the hair growth condition of the depilated area on the back of the mice in each experimental group; Figure 4 is a statistical diagram of the hair growth condition of the depilated area on the back of the mice in each experimental group; Figure 5 is the cytotoxicity result of different microneedle materials on HDPCs; Figure 6 is the cytotoxicity result of different microneedle materials on HFSCs.

[0026] The reference signs are: microneedle 1, needle tip segment 11, needle root segment 12, patch 2. DETAILED DESCRIPTION

[0027] The present application will be further described below in conjunction with examples. Those skilled in the art will be able to implement the present application based on these descriptions. In addition, the examples of the present application involved in the following descriptions are generally only examples of a part of the present application, rather than all examples. Therefore, all other examples obtained by those skilled in the art based on the examples in the present application without making creative efforts shall belong to the protection scope of the present application.

[0028] For AGA, finasteride is a drug with certain curative effect, which belongs to 5alpha-reductase inhibitor, mainly acts on hair papilla cells, inhibits the conversion of testosterone to dihydrotestosterone (DHT), improves the symptoms of hair loss, promotes the growth of hair, and thus achieves the effect of treating AGA. However, finasteride is limited by its mechanism of action, and its existing curative effect is highly bottlenecked, so it is necessary to explore safer and more efficient intervention means to improve the therapeutic effect of finasteride.

[0029] Therefore, the present application provides a microneedle patch based on a hierarchical targeting mechanism, which combines Figure 1 and Figure 2As shown, the microneedle patch includes microneedles 1 and a patch 2, the material forming the microneedle body includes a water-soluble polymer, the microneedle body includes a needle tip section 11 and a needle root section 12, the needle tip section 11 is loaded with finasteride, and the needle root section 12 is loaded with quercetin.

[0030] The present application promotes the targeting of finasteride to hair papilla cells and inhibits the conversion of testosterone to DHT in hair papilla cells by the quercetin provided in the needle root section, thereby improving the therapeutic effect of finasteride on AGA.

[0031] The present application forms a microneedle patch with a hierarchical targeting mechanism by the hierarchical targeting of different cells by finasteride provided in the needle tip section and quercetin provided in the needle root section. The principle of the hierarchical targeting mechanism promoting therapeutic effect is that after the microneedle pierces the skin, the water-soluble polymer in the needle tip section dissolves, releasing finasteride to target hair papilla cells and inhibit the conversion of testosterone to DHT in hair papilla cells. During the targeting of hair papilla cells by finasteride, quercetin released from the needle root section gradually targets endothelial cells, promoting the proliferation and migration of endothelial cells, thereby strengthening the formation of a highly branched capillary network by endothelial cells acting on hair papilla cells to meet their metabolic needs, ultimately enhancing the targeting effect of finasteride. The needle root section is relatively thicker than the needle tip section and pierces the skin to a shallower depth, contacting less liquid, so the drug release rate of the needle root section is relatively slower than that of the needle tip section. The present application gradually acts on endothelial cells by relatively slow-release quercetin, thereby improving the therapeutic effect of finasteride on AGA.

[0032] As a preferred embodiment of the above-mentioned microneedle patch, the quercetin is loaded by a metal-organic framework and then loaded by the water-soluble polymer to form the needle root section.

[0033] By loading the metal-organic framework, the quercetin is released more slowly, which can more continuously improve the metabolism of hair papilla cells during the targeting of finasteride to hair papilla cells and the inhibition of the conversion of testosterone to DHT in hair papilla cells.

[0034] After the microneedle pierces the skin, the tip segment dissolves faster than the root segment because of its sharp tip, so the microneedle releases quercetin in the root segment relatively slowly, and after being loaded by the metal organic framework, more quercetin can be absorbed by endothelial cells in the middle and late stages of finasteride action. In the middle and late stages of finasteride acting on the hair papilla cells, the process of converting endogenous testosterone to DHT in the metabolism of hair papilla cells has been inhibited, and promoting the proliferation of endothelial cells and the metabolism of hair papilla cells is more conducive to the inhibition of the conversion of endogenous testosterone to DHT. The enhanced metabolism of hair papilla cells is conducive to the proliferation and differentiation of hair follicle stem cells by secreting Wnt and other signal molecules, thereby improving the symptoms of hair loss and promoting hair growth.

[0035] Further preferably, the metal organic framework is ZIF-8.

[0036] By loading quercetin with ZIF-8 and then setting it in the root segment of the microneedle, the slow-release effect of the root segment on quercetin is achieved.

[0037] As a preferred microneedle patch, the water-soluble polymer is selected from one or more of hyaluronic acid, sodium alginate, and dextran.

[0038] As a preferred microneedle patch, the finasteride loaded in the tip segment is a liposome.

[0039] It can be understood that the patch structure in the above-mentioned microneedle patch of the present application can be obtained by existing technology, which is a conventional structure of a drug-loaded microneedle system.

[0040] In addition, the present application provides a preparation method of a microneedle patch based on a hierarchical targeting mechanism, which comprises the following steps: Step S1, a step of preparing finasteride liposomes to form finasteride liposomes; Step S2, a step of preparing a metal organic framework and loading quercetin to form a quercetin-loaded metal organic framework Qu@MOF; Step S3, mixing the finasteride liposomes with the hyaluronic acid solution to obtain the tip segment material of the microneedle, and mixing the Qu@MOF with the hyaluronic acid solution to obtain the root segment material of the microneedle; Step S4, sequentially pouring the tip segment material and the root segment material into a microneedle mold, drying, and forming a microneedle.

[0041] As a preferred preparation method, the step of preparing finasteride liposomes comprises: Mix 10-15 mg glycerin monostearate and 5-10 mg finasteride, heat to 70-90℃ until glycerin monostearate melts, and dissolve 10-30 μL oleic acid in the lipid matrix to obtain an organic phase; mix soybean lecithin and emulsifier in 10-25 mL distilled water, heat to 70-90℃ to obtain an aqueous phase, wherein the concentration of the soybean lecithin is 0.5-2 g / mL, and the concentration of the emulsifier is 2-4 g / mL; mix the organic phase and the aqueous phase, stir to form a primary emulsion, and ultrasonically treat to form a fine emulsion; add the fine emulsion to 20-35 mL water containing 2-4 g / mL emulsifier, cool, and obtain a finasteride liposome dispersion.

[0042] Further, the emulsifier is preferably Tween-80.

[0043] As a preferred embodiment of the preparation method, the step of preparing the metal organic framework and loading quercetin comprises: Disperse 2-methylimidazole and quercetin in methanol to obtain a 2-methylimidazole mixture; mix the 2-methylimidazole mixture with a zinc nitrate solution, stir, and collect white solids to obtain a metal organic framework loaded with quercetin.

[0044] As a preferred embodiment of the preparation method, the drying temperature is 30-40℃, and the drying time is 4-8 hours.

[0045] Example 1: Synthesis of finasteride nanoliposomes (FIN-NLC) Mix 10-15 mg glycerin monostearate and 10 mg finasteride, heat to 80℃ until glycerin monostearate melts, and dissolve 20 μL oleic acid in the lipid matrix to obtain an organic phase. Mix soybean lecithin and Tween-80 in 25 mL distilled water to obtain an aqueous phase, wherein the concentration of the soybean lecithin is 1 g / mL, and the concentration of the Tween-80 is 3 g / mL. Heat to 80℃ to obtain the aqueous phase. Mix the organic phase and the aqueous phase under magnetic stirring at 800 rpm, and stir for 60 min to form a primary emulsion. Then ultrasonically treat the primary emulsion at a power of 500 w for 20 times, 3 s working and 2 s stopping each time, to obtain a fine emulsion. Precool water containing 3 g / mL emulsifier to 4℃, and then add the obtained fine emulsion to 35 mL water containing 3 g / mL emulsifier, and cool at 4℃ to obtain a dispersion of finasteride nanoliposomes FIN-NLC. Dilute the dispersion with deionized water to obtain a FIN-NLC liquid with a finasteride concentration of 50 μg / mL, for standby use.

[0046] Example 2: Synthesis of quercetin-loaded ZIF-8 (Qu@ZIF-8) Zn(NO3)2*6H2O (150 mg, 0.5 mmol) was dissolved in H2O (5 mL). And 2-mIM (2-methylimidazole, 330 mg, 4 mmol) was dispersed in MeOH (10 mL). The two solutions were quickly poured together under magnetic stirring at 500 rpm. After 10 min of magnetic stirring, the white solid product was collected by centrifugation at 8000 rpm for 20 min, washed with MeOH several times, and the residual solvent was removed under vacuum at 60 °C to obtain Qu@ZIF-8, i.e. quercetin-loaded ZIF-8. Qu@ZIF-8 was added to water to prepare a Qu@ZIF-8 solution with a quercetin concentration of 500 pg / mL, ready for use.

[0047] Example 3 Preparation of a microneedle patch A microneedle patch based on a hierarchical targeting mechanism was prepared, including the following steps: Step S1, take 100 mL of FIN-NLC solution obtained in Example 1 (the concentration of finasteride is 50 pg / mL), add 5 g of hyaluronic acid, stir and mix to obtain the needle tip segment material of the microneedle.

[0048] Step S2, take 100 mL of Qu@ZIF-8 solution obtained in Example 2 (the concentration of quercetin is 500 pg / mL), add 5 g of hyaluronic acid, stir and mix to obtain the needle root segment material of the microneedle.

[0049] Step S3, pour the needle tip segment material into the microneedle mold (polydimethylsiloxane (PDMS) mold), dry in an oven at 37 °C for 3 hours to semi-dry, then continue to pour the needle root segment material into the microneedle mold, and dry the sample again in an oven at 37 °C for 8 h to form the microneedle solid structure, the height ratio of the needle tip segment h1 to the needle root segment h2 is 1:2 (300 pm:600 pm). After drying, demold to obtain the microneedle patch FZQ-MN.

[0050] Example 4 Preparation of a microneedle patch A microneedle patch based on a hierarchical targeting mechanism was prepared, including the following steps: Step S1, take 100 mL of FIN-NLC solution obtained in Example 1 (the concentration of finasteride is 50 pg / mL), add 5 g of hyaluronic acid, stir and mix to obtain the needle tip segment material of the microneedle.

[0051] Step S2, dissolve quercetin in deionized water to obtain a quercetin solution with a concentration of 500 pg / mL, take 100 mL of the quercetin solution, add 5 g of hyaluronic acid to it, stir and mix to obtain the needle root segment material of the microneedle.

[0052] Step S3, pour the tip segment material into the microneedle mold (polydimethylsiloxane (PDMS) mold), dry in the oven at 37°C for 3 hours to semi-dry, then continue to pour the root segment material into the microneedle mold, and dry the sample again in the oven at 37°C for 8 h to form a microneedle solid structure, the height ratio h1: h2 of the tip segment of the microneedle to the root segment is 1:2 (300 μm: 600 μm). After drying, demold to obtain microneedle patch B-MN.

[0053] Example 5 Preparation of microneedle patch A microneedle patch based on a hierarchical targeting mechanism was prepared, including the following steps: Step S1, take 100 mL of FIN-NLC liquid obtained in Example 1 (the concentration of finasteride is 50 μg / mL), add 100 mL of hyaluronic acid solution (the concentration of hyaluronic acid is 5 g / mL), stir and mix to obtain the tip segment material of the microneedle.

[0054] Step S2, take 100 mL of Qu@ZIF-8 liquid obtained in Example 2 (the concentration of quercetin is 500 μg / mL), add 100 mL of hyaluronic acid solution (the concentration of hyaluronic acid is 5 g / mL), stir and mix to obtain the root segment material of the microneedle.

[0055] Step S3, pour the tip segment material into the microneedle mold (polydimethylsiloxane (PDMS) mold), dry in the oven at 37°C for 3 hours to semi-dry, then continue to pour the root segment material into the microneedle mold, and dry the sample again in the oven at 37°C for 8 h to form a microneedle solid structure, the height ratio h1: h2 of the tip segment of the microneedle to the root segment is 1:2 (300 μm: 600 μm). After drying, demold to obtain microneedle patch B-MN.

[0056] Comparative Example 1 A smearing ointment containing finasteride and quercetin was prepared, which was different from Example 3 in that the drug delivery system was different. The preparation steps of this comparative example are as follows: Step S1, take 100 mL of FIN-NLC liquid obtained in Example 1 (the concentration of finasteride is 50 μg / mL), add 5 g of hyaluronic acid, stir and mix to obtain the tip segment material.

[0057] Step S2, take 100 mL of Qu@ZIF-8 liquid obtained in Example 2 (the concentration of quercetin is 500 μg / mL), add 5 g of hyaluronic acid, stir and mix to obtain the root segment material.

[0058] Step S3, mix the tip segment material and the root segment material according to the volume ratio of 1:26, then dry in the oven at 37°C for 6 h to obtain an ointment, which is the smearing ointment FZQ.

[0059] Comparative Example 2 A single-targeting microneedle patch containing only finasteride was prepared, which was different from Example 3 in that the FIN-NLC liquid in the material of the needle tip segment was replaced with an equal volume of Qu@ZIF-8 liquid. The following steps were included: Step S1, take 100 mL of FIN-NLC liquid obtained in Example 1 (the concentration of finasteride is 50 μg / mL), add 5 g of hyaluronic acid, stir and mix evenly to obtain the material of the needle tip segment of the microneedle.

[0060] Step S2, pour the needle tip segment material into the microneedle mold (polydimethylsiloxane (PDMS) mold) until full, then dry the sample in an oven at 37°C for 14 h to form the microneedle solid structure, and the total height of the microneedle is 900 μm. After drying, demold to obtain the microneedle patch F-MN.

[0061] Comparative Example 3 A single-targeting microneedle patch containing only quercetin was prepared, which was different from Example 3 in that the FIN-NLC liquid in the material of the needle tip segment was replaced with an equal volume of Qu@ZIF-8 liquid. The following steps were included: Step S1, take 100 mL of Qu@ZIF-8 liquid obtained in Example 2 (the concentration of quercetin is 500 μg / mL), add 5 g of hyaluronic acid, stir and mix evenly to obtain the material of the needle tip segment of the microneedle.

[0062] Step S3, pour the needle tip segment material into the microneedle mold until full, then dry the sample in an oven at 37°C for 14 h to form the microneedle solid structure, and the total height of the microneedle is 900 μm. After drying, demold to obtain the microneedle patch ZQ-MN.

[0063] Performance characterization (1) Male C57BL / 6 mice were adaptively fed for 7 days. Subsequently, these mice were randomly divided into 8 experimental groups, each with 5 mice. After the back of the 8 experimental groups was shaved, except for the healthy group (Healthy group), the other mice were smeared with HT on the back of the depilated area to induce the formation of an AGA model. After the AGA model was completed, the 8 experimental groups were treated for a total of 21 days as follows: Healthy group: no treatment; A-MN group, referred to as FZQ-MN group: smearing HT on the back of the depilated area every day, and using the FZQ-MN patch prepared in Example 3 for treatment; B-MN group: smearing HT on the back of the depilated area every day, and using the B-MN patch prepared in Example 4 for treatment; FZQ group: smearing HT on the back of the depilated area every day, and using the smearing ointment FZQ prepared in Comparative Example 1 for treatment, and the amount of smearing ointment used is 2 times the weight of the microneedle used on the depilated area; F-MN group: HT was applied to the depilation area of the back every day, and F-MN patches prepared using Comparative Example 2 were attached for treatment; ZQ-MN group: HT was applied to the depilation area of the back every day, and ZQ-MN patches prepared using Comparative Example 3 were attached for treatment; Minoxidil group: HT was applied to the depilation area of the back every day, and 2% minoxidil was used for treatment, 2 mL per time; AGA group: HT was applied to the depilation area of the back every day, and blank microneedle patches without drug were attached for treatment, and the blank microneedles were molded using a 5% hyaluronic acid solution according to the method of Example 3.

[0064] The hair growth conditions of the depilation area of the back of the 8 experimental groups were photographed on the 1st day, 4th day, 7th day, 11th day, 14th day, 17th day and 21st day of the experiment, and the results are shown in Figure 3 and Figure 4 , Figure 3 is a photograph of the hair growth condition record of the depilation area of the back, Figure 4 is a corresponding hair growth condition statistical chart.

[0065] From Figure 3 and Figure 4 It can be seen that the finasteride is arranged on the needle tip segment of the microneedle, and the quercetin is arranged on the needle root segment, different cells are targeted by the finasteride and the quercetin, the microneedle patch with a hierarchical targeting mechanism is formed, and the hair growth effect is better than that of the F-MN group and the ZQ-MN group of a single drug, and the effect is also better than that of the Minoxidil group of the traditional minoxidil drug.

[0066] The principle of the analysis lies in that after the microneedle pierces the skin, the water-soluble polymer of the needle tip segment dissolves to release the finasteride to target the hair papilla cells, and then inhibit the conversion of testosterone into DHT in the hair papilla cells. In the process of targeting the hair papilla cells by the finasteride, the quercetin released by the needle root segment gradually targets the endothelial cells to promote the proliferation and migration activity of the endothelial cells, so as to strengthen the effect of the endothelial cells on the hair papilla cells to form a highly branched capillary network, maintain the metabolic demand of the hair papilla cells, and finally play a role in strengthening the targeting effect of the finasteride. Therefore, the microneedle patch with a hierarchical targeting mechanism of the present application is higher than the AGA effect of the microneedle patch of a single finasteride drug, and is also higher than the AGA effect of the finasteride and quercetin combined application of the smearing type drug.

[0067] From the comparison between the FZQ-MN group and the B-MN group, it can be seen that after the metal organic framework is loaded with quercetin, the metal organic framework releases the quercetin (FZQ-MN group), which is better than the relatively slow release of the needle root segment (B-MN group), and the hair growth promoting effect is better (FZQ-MN group).

[0068] The principle of the analysis can be that after the micro-needles pierce the skin, the tip segment dissolves faster than the root segment because it is a sharp tip, so the micro-needle releases the quercetin of the root segment relatively slowly, and after being loaded by the metal organic framework, more quercetin can be absorbed by the endothelial cells in the middle and late stages of the action of finasteride. In the middle and late stages of the action of finasteride on the hair papilla cells, the process of converting endogenous testosterone to DHT in the metabolism of the hair papilla cells has been inhibited, and promoting the proliferation of endothelial cells and the metabolism of hair papilla cells is more conducive to the inhibition of the conversion of endogenous testosterone to DHT.

[0069] The cytotoxicity of the micro-needles provided by the application to HDPCs and HFSCs was detected by CCK-8 method. HDPCs and HFSCs were seeded in a 96-well culture plate at a density of 5×10 3 μg / mL of FIN, ZIF-8, Qu@ZIF-8 (n=4) were added, and after 24h of incubation, 10μL of CCK-8 solution was added to each well, and the plate was incubated in a CO2 incubator. The absorbance of each well at 450nm was measured by an enzyme-labeled instrument. The cytotoxicity results of HDPCs and HFSCs are shown in Figure 5 and Figure 6 .

[0070] As can be seen from Figure 5 and Figure 6 , when the FIN-NLC is 5μg / mL and the Qu@ZIF-8 is 50μg / mL, HDPCs and HFSCs can have better cell viability.

[0071] The raw materials and equipment used in the application are conventional raw materials and equipment in the art unless otherwise specified; the methods used in the application are conventional methods in the art unless otherwise specified.

[0072] The above is only a preferred embodiment of the application, and does not limit the application in any way. Any simple modification, change and equivalent transformation of the above embodiment based on the technical essence of the application still falls within the protection scope of the technical solution of the application.

Claims

1. A microneedle patch based on a hierarchical targeting mechanism, comprising microneedles and a patch, characterized in that: The material forming the microneedle body includes a water-soluble polymer, and the microneedle body includes a tip segment and a root segment. The tip segment is loaded with finasteride, and the root segment is loaded with quercetin.

2. The microneedle patch based on a hierarchical targeting mechanism as described in claim 1, characterized in that: The quercetin, after being loaded onto a metal-organic framework, is then loaded onto a water-soluble polymer to form the needle root segment.

3. A microneedle patch based on a hierarchical targeting mechanism as described in claim 1, characterized in that: The metal-organic framework is ZIF-8.

4. A microneedle patch based on a hierarchical targeting mechanism as described in claim 1, characterized in that: The water-soluble polymer is selected from one or more of hyaluronic acid, sodium alginate, and dextran.

5. A microneedle patch based on a hierarchical targeting mechanism as described in claim 1, characterized in that: The finasteride loaded on the needle tip is a liposome.

6. The method for preparing a microneedle patch based on a hierarchical targeting mechanism as described in claim 1, characterized in that: Includes the following steps: Step S1: The step of preparing finasteride liposomes, forming finasteride liposomes; Step S2, the step of preparing metal-organic framework and loading quercetin, to form a quercetin-loaded metal-organic framework Qu@MOF; Step S3: Mix finasteride liposomes with hyaluronic acid solution to obtain the tip segment material of microneedles; mix Qu@MOF with hyaluronic acid solution to obtain the root segment material of microneedles; Step S4: Pour the needle tip material and needle root material into the microneedle mold in sequence, dry them, and form microneedles.

7. The preparation method according to claim 6, characterized in that: The step of preparing finasteride liposomes includes: Mix 10-15 mg of glyceryl monostearate and 5-10 mg of finasteride, heat to 70-90°C until the glyceryl monostearate melts, and dissolve 10-30 μL of oleic acid in the lipid matrix to obtain the organic phase; add soybean lecithin and emulsifier to 10-25 mL of distilled water, heat to 70-90°C to obtain the aqueous phase, wherein the concentration of soybean lecithin is 0.5-2 g / mL and the concentration of emulsifier is 2-4 g / mL; mix the organic phase and the aqueous phase, stir to form a primary emulsion, and sonicate to form a fine emulsion; add the fine emulsion to 20-35 mL of water containing 2-4 g / mL of emulsifier, cool, and obtain a finasteride liposome dispersion.

8. The preparation method according to claim 6, characterized in that: The steps for preparing the metal-organic framework and loading quercetin include: 2-Methylimidazole and quercetin were dispersed in methanol to obtain a 2-methylimidazole mixture; the 2-methylimidazole mixture was mixed with zinc nitrate solution, stirred, and the white solid was collected to obtain the metal-organic framework loaded with quercetin.

9. The preparation method according to claim 6, characterized in that: The drying temperature is 30~40℃, and the drying time is 4~8 hours.

10. The application of the microneedle patch as described in any one of claims 1 to 5, or the microneedle patch prepared by the preparation method as described in any one of claims 6 to 9, in the treatment of androgenetic alopecia.

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