A method for preparing an inflammation-responsive microneedle patch for atopic dermatitis

Abstract

The present invention discloses a method for preparing an inflammation-responsive microneedle patch for atopic dermatitis, which adopts a method of polydimethylsiloxane (PDMS) mold replication, adopts biocompatible and degradable raw materials 1 and raw materials 2, and forms a microneedle patch by ultraviolet light curing and drying in two steps. The inflammation-responsive microneedle patch for atopic dermatitis prepared by the present invention has a matrix metalloproteinase degradation site, which can be degraded by inflammatory mediators-matrix metalloproteinases, and its degradation rate increases with the increase of matrix metalloproteinase concentration. The drug release process of the microneedle patch is closely related to the degradation process, and the inflammation-responsive release of the drug can be achieved by the degradation of matrix metalloproteinases. The preparation method of the inflammation-responsive microneedle patch for atopic dermatitis is simple and easy, has low requirements for instruments and equipment, has high production efficiency, can be mass-produced by multiple batches of templates, and has good application prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical materials, and in particular to an inflammation-responsive microneedle patch for atopic dermatitis and a preparation method thereof. Background Art

[0002] Atopic dermatitis is a chronic, recurrent, inflammatory skin disease, with recurrent chronic eczematous rashes as the main clinical manifestations, accompanied by significant skin dryness and itching. It is also characterized by epidermal barrier defects and immune hyperresponsiveness to allergens, which brings a series of serious psychological and physiological burdens to patients and their families. In addition, although many methods have been proven to be effective, their effectiveness in relieving atopic dermatitis symptoms and reducing recurrence rates is still limited. Therefore, there is an urgent need to develop other methods to better meet the complex needs of atopic dermatitis treatment.

[0003] Microneedles are a new type of effective drug delivery tool proposed in recent years. They can penetrate the epidermis in a non-invasive and non-infectious way. Due to their microscopic size, microneedles can cross blood vessels and peripheral nerves to deliver drugs transdermally without causing serious consequences such as vascular rupture and pain. In addition, by changing the raw materials of microneedles, they can also achieve sustained release and controlled release, achieving long-term effective treatment effects.

[0004] CN107405301A relates to a microneedle patch composition comprising one or more microneedles, each of which comprises: (a) a tapered tip portion containing a therapeutically active ingredient dispersed in a matrix of a biodegradable polymer, which is capable of providing sustained release of the therapeutically active ingredient after the one or more microneedles are inserted into the skin, and (b) a dissolvable microneedle backing layer portion containing a water-soluble polymer covering the tip portion, the one or more microneedles being attached to a surface of a removable substrate and extending from the surface.

[0005] WO02 / 046193 relates to a microneedle device (10) and a method of manufacture, provided for transporting molecules or energy across or into a biological barrier, such as the skin. The device may include one or more microneedles formed from a first material and a second material, wherein the second material is dispersed throughout the first material or forms a portion of the microneedle. The first material is preferably a polymer. The second material may be a pore former, a structural component, a biosensor, or a molecule for release, such as a drug. The device may also include a substrate and a plurality of microneedles extending from the substrate, wherein the microneedles have a beveled or tapered tip portion, a longitudinally extending external channel, or both. The method of manufacturing these devices includes providing a mold having a plurality of microrecesses that define the surface of the microneedles; filling the microrecesses with a first molding material; and molding the material to form the microneedles.

[0006] Although some microneedle patches have been studied and developed in the above-mentioned prior art, there are still some problems in the prior art, such as the lack of microneedle patches specifically for atopic dermatitis, the insufficient degradation rate of the microneedle patches and their remaining in the body as foreign matter, and the inability of the microneedle patches to successfully penetrate the thickened skin of atopic dermatitis patients deep into the epidermis due to insufficient mechanical properties, resulting in poor treatment effects.

[0007] Therefore, the present invention adopts the method of polydimethylsiloxane (PDMS) mold replication, adopts biocompatible and degradable methacrylate gelatin and sodium hyaluronate, and forms microneedle patch by ultraviolet light curing and drying two steps. The inflammatory responsive microneedle patch for atopic dermatitis prepared by the present invention has matrix metalloproteinase degradation site, can be degraded by inflammatory mediator-matrix metalloproteinase, and its degradation rate increases with the increase of matrix metalloproteinase concentration. The drug release process of the microneedle patch is closely related to the degradation process, and the inflammatory responsive release of the drug can be achieved by the degradation of matrix metalloproteinase. The preparation method of the inflammatory responsive microneedle patch for atopic dermatitis is simple and easy, has low requirements for instruments and equipment, has high production efficiency, can realize mass production by multiple batches of templates, has good application prospects, and is of great significance for alleviating the symptoms of atopic dermatitis. Summary of the invention

[0008] The technical problem to be solved by the present invention is to provide an inflammation-responsive microneedle patch for atopic dermatitis and a preparation method thereof in view of the deficiencies of the above-mentioned prior art.

[0009] In order to achieve the above technical objectives, the technical solution adopted by the present invention is:

[0010] The present invention provides a method for preparing an inflammation-responsive microneedle patch for atopic dermatitis, comprising the following steps:

[0011] 1) Preparation of microneedle raw material solution:

[0012] The raw material 1, the therapeutic drug and 1% 2-hydroxy-2-methylpropiophenone were uniformly mixed to obtain a microneedle raw material solution 1; the raw material 2 was dissolved in pure water, and the microneedle raw material solution 2 was obtained after centrifugation to remove bubbles, and stored at 4°C for later use;

[0013] 2) Preparation of inflammation-responsive microneedle patches:

[0014] Add microneedle raw material solution 1 to the PDMS microneedle small mold, and evenly fill the needle body of the small mold with the solution by centrifugation or vacuuming, scrape off the excess solution and solidify it by ultraviolet light. Add microneedle raw material solution 2 as the inflammation-responsive microneedle patch substrate, and demold after drying to obtain a microneedle array. Then add microneedle raw material solution 2 to the PDMS microneedle large mold, and evenly fill the needle body of the large mold with the solution by centrifugation or vacuuming, scrape off the excess solution, position the above-mentioned microneedle array and press it into the large mold, and then dry and demold to obtain an inflammation-responsive microneedle patch.

[0015] Furthermore, in step (1), the microneedle raw material 1 is selected from methacrylate gelatin. The therapeutic drug is selected from one or more of triamcinolone acetonide, mometasone furoate and dexamethasone, with a concentration of 20-25 mg / mL.

[0016] Furthermore, in step (2), the microneedle raw material 2 is selected from one or more of sodium hyaluronate, polyvinyl alcohol, and polyvinyl pyrrolidone.

[0017] Furthermore, in step (2), in the two microneedle molds of different sizes, the needle tip length of the large microneedle mold is 0.4-1.5 mm, and the needle tip length of the small microneedle mold is 0.2-1 mm.

[0018] Furthermore, in step (2), of the two microneedle molds of different sizes, the bottom radius of the large microneedle mold is 100-600 μm, and the bottom radius of the small microneedle mold is 100-300 μm.

[0019] Furthermore, in step (2), 1% 2-hydroxy-2-methylpropiophenone is added to the microneedle raw material solution 1, and then cured by ultraviolet light.

[0020] Furthermore, the microneedle raw material solution 1 needs to be kept away from light after adding 1% 2-hydroxy-2-methylpropiophenone.

[0021] Furthermore, the drying temperature is lower than 37°C.

[0022] The present invention also provides an inflammation-responsive microneedle for atopic dermatitis prepared by the above-mentioned preparation method, which has a matrix metalloproteinase degradation site and can be degraded by the inflammatory mediator-matrix metalloproteinase, and its degradation rate increases with the increase of the concentration of matrix metalloproteinase. The drug release process of the microneedle patch is closely related to the degradation process, and the inflammation-responsive release of the drug can be achieved through the degradation of matrix metalloproteinase. The preparation method of the inflammation-responsive microneedle patch for atopic dermatitis is simple and easy, has low requirements for instruments and equipment, has high production efficiency, can be mass-produced through multiple batches of templates, has good application prospects, and is of great significance for alleviating the symptoms of atopic dermatitis.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The method for preparing an inflammation-responsive microneedle patch for atopic dermatitis provided by the present invention comprises adding a microneedle raw material solution 1 to a small PDMS microneedle mold, and after the solution is uniformly filled into the needle body of the small mold by centrifugation or vacuuming, the excess solution is scraped off and cured by ultraviolet light. A microneedle raw material solution 2 is added to serve as the inflammation-responsive microneedle patch substrate, and the microneedle array is obtained by demolding after drying. The microneedle raw material solution 2 is then added to a large PDMS microneedle mold, and after the solution is uniformly filled into the needle body of the large mold by centrifugation or vacuuming, the excess solution is scraped off, the above-mentioned microneedle array is positioned and pressed into the large mold, and then the microneedle array is dried and demolded to obtain an inflammation-responsive microneedle patch. The preparation process is simple and easy, and the reaction conditions are low.

[0025] The inflammatory responsive microneedle patch for atopic dermatitis prepared by the present invention has a matrix metalloproteinase degradation site, which can be degraded by the inflammatory mediator-matrix metalloproteinase, and its degradation rate increases with the increase of the concentration of matrix metalloproteinase. The drug release process of the microneedle patch is closely related to the degradation process, and the inflammatory responsive release of the drug can be achieved through the degradation of matrix metalloproteinase, which is of great significance for alleviating the symptoms of atopic dermatitis.

[0026] The preparation method of the double-layer microneedle patch provided by the present invention utilizes two different raw material solutions to fill and demold large and small molds to form a multi-layer microneedle structure, thereby achieving the encapsulation of anti-inflammatory drugs. When the microneedles act on the skin, the outer needle tip components are dissolved under the action of tissue fluid, while the inner needle tip components are released, thereby achieving the effect of inflammation-responsive release. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a physical picture of an inflammation-responsive microneedle patch for atopic dermatitis of the present invention;

[0028] Figure 2 This is a degradation curve of an inflammation-responsive microneedle patch for atopic dermatitis of the present invention;

[0029] Figure 3 This is a drug release curve of an inflammation-responsive microneedle patch for atopic dermatitis of the present invention. DETAILED DESCRIPTION

[0030] In order to enable those skilled in the art to better understand the technical solution of the present invention, the embodiments of the present invention are further described in detail below with reference to the accompanying drawings.

[0031] The experimental methods used in the following examples are conventional methods unless otherwise specified, and the reagents, methods and equipment used are conventional reagents, methods and equipment in the art unless otherwise specified.

[0032] In the description of this specification, the description with reference to the terms "one embodiment", "another embodiment", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment are included in at least one embodiment of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and the features of different embodiments or examples without contradiction.

[0033] The present invention provides a method for preparing an inflammation-responsive microneedle patch for atopic dermatitis, comprising the following steps:

[0034] Step 1: Preparation of microneedle raw material solution:

[0035] The raw material 1, the therapeutic drug and 1% 2-hydroxy-2-methylpropiophenone are uniformly mixed to obtain a microneedle raw material solution 1;

[0036] Raw material 2 was dissolved in pure water, and the bubbles were removed by centrifugation to obtain microneedle raw material solution 2, which was stored at 4°C for future use;

[0037] Step 2: Preparation of inflammation-responsive microneedle patches:

[0038] Add microneedle raw material solution 1 to the PDMS microneedle small mold, and evenly fill the needle body of the small mold with the solution by centrifugation or vacuuming, scrape off the excess solution and solidify it by ultraviolet light. Add microneedle raw material solution 2 as the inflammation-responsive microneedle patch substrate, and demold after drying to obtain a microneedle array. Then add microneedle raw material solution 2 to the PDMS microneedle large mold, and evenly fill the needle body of the large mold with the solution by centrifugation or vacuuming, scrape off the excess solution, position the above-mentioned microneedle array and press it into the large mold, and then dry and demold to obtain an inflammation-responsive microneedle patch.

[0039] In the first step, the microneedle raw material 1 is selected from methacrylate gelatin. The therapeutic drug is selected from one or more of triamcinolone acetonide, mometasone furoate and dexamethasone.

[0040] In the first step, the microneedle raw material 2 is selected from one or more of sodium hyaluronate, polyvinyl alcohol, and polyvinyl pyrrolidone.

[0041] In the second step, in the two microneedle molds of different sizes, the needle tip length of the large microneedle mold is 0.4-1.5 mm, and the needle tip length of the small microneedle mold is 0.2-1 mm.

[0042] In the second step, of the two microneedle molds of different sizes, the bottom radius of the large microneedle mold is 100-600 μm, and the bottom radius of the small microneedle mold is 100-300 μm.

[0043] In the second step, 1% 2-hydroxy-2-methylpropiophenone is added to the microneedle raw material solution 1, and then cured by ultraviolet light.

[0044] After adding 1% 2-hydroxy-2-methylpropiophenone, the microneedle raw material solution 1 should be placed away from light.

[0045] The drying temperature is lower than 37°C.

[0046] The method for preparing an inflammation-responsive microneedle patch for atopic dermatitis provided by the present invention comprises adding a microneedle raw material solution 1 to a small PDMS microneedle mold, and after the solution is uniformly filled into the needle body of the small mold by centrifugation or vacuuming, the excess solution is scraped off and cured by ultraviolet light. A microneedle raw material solution 2 is added to serve as the inflammation-responsive microneedle patch substrate, and the microneedle array is obtained by demolding after drying. The microneedle raw material solution 2 is then added to a large PDMS microneedle mold, and after the solution is uniformly filled into the needle body of the large mold by centrifugation or vacuuming, the excess solution is scraped off, the above-mentioned microneedle array is positioned and pressed into the large mold, and then the microneedle array is dried and demolded to obtain an inflammation-responsive microneedle patch. The preparation process is simple and easy, and the reaction conditions are low.

[0047] The inflammatory responsive microneedle patch for atopic dermatitis prepared by the present invention has a matrix metalloproteinase degradation site, which can be degraded by the inflammatory mediator-matrix metalloproteinase, and its degradation rate increases with the increase of the concentration of matrix metalloproteinase. The drug release process of the microneedle patch is closely related to the degradation process, and the inflammatory responsive release of the drug can be achieved through the degradation of matrix metalloproteinase, which is of great significance for alleviating the symptoms of atopic dermatitis.

[0048] The following are examples:

[0049] Example 1: Inflammation-responsive microneedle patch for atopic dermatitis and preparation method thereof

[0050] An inflammatory responsive microneedle patch for atopic dermatitis based on methacrylate gelatin and sodium hyaluronate, the preparation process comprising the following steps:

[0051] Step 1: Prepare microneedle raw material solution:

[0052] The methacrylate gelatin containing the therapeutic drug (triamcinolone acetonide, 20 mg / mL) with a mass fraction of 30% was uniformly mixed with 1% 2-hydroxy-2-methylpropiophenone to obtain a microneedle raw material solution 1, which was stored in the dark for later use; sodium hyaluronate was dissolved in pure water to prepare a 22.5% sodium hyaluronate solution, and the microneedle raw material solution 2 was obtained after centrifugation to remove bubbles, and stored at 4° C. for later use;

[0053] Step 2: Preparation of inflammation-responsive microneedle patches:

[0054] The microneedle raw material solution 1 is added to the PDMS microneedle small mold in a slightly hot state, and centrifuged at 5000rpm for 1min to allow the solution to evenly fill the needle body of the small mold. The excess solution is then scraped off with a coverslip, and the solution is solidified by ultraviolet light for 30s. Microneedle raw material solution 2 is added to serve as the inflammation-responsive microneedle patch substrate, and the microneedle array is obtained after being demolded at 37°C for 12h. The microneedle raw material solution 2 is then added to the PDMS microneedle large mold, and centrifuged at 4°C, 5000rpm for 1min to allow the solution to evenly fill the needle body of the large mold. The excess solution is then scraped off with a coverslip, and the above-mentioned microneedle array is positioned and pressed into the large mold, and then dried at 37°C for 6h to obtain an inflammation-responsive microneedle patch. Figure 1 shown.

[0055] Example 2: Inflammation-responsive microneedle patch for atopic dermatitis and preparation method thereof

[0056] An inflammatory responsive microneedle patch for atopic dermatitis based on methacrylate gelatin and polyvinyl pyrrolidone, the preparation process comprising the following steps:

[0057] Step 1: Prepare microneedle raw material solution:

[0058] The methacrylate gelatin containing the therapeutic drug (dexamethasone, 20 mg / mL) with a mass fraction of 20% was uniformly mixed with 1% 2-hydroxy-2-methylpropiophenone to obtain a microneedle raw material solution 1, which was stored in the dark for later use; polyvinyl pyrrolidone was dissolved in pure water to prepare a 22.5% polyvinyl pyrrolidone solution, and the microneedle raw material solution 2 was obtained after centrifugation to remove bubbles, and stored at 4° C. for later use;

[0059] Step 2: Preparation of inflammation-responsive microneedle patches:

[0060] The microneedle raw material solution 1 was added to the PDMS microneedle small mold in a slightly hot state, and the solution was evenly filled into the needle body of the small mold by centrifugation at 5000rpm for 1min, and the excess solution was scraped off with a cover glass, and it was solidified by ultraviolet light for 30s. Microneedle raw material solution 2 was added to serve as the inflammation-responsive microneedle patch substrate, and the microneedle array was obtained after being demolded at 37℃ for 12h. Then, the microneedle raw material solution 2 was added to the PDMS microneedle large mold, and the solution was evenly filled into the needle body of the large mold by centrifugation at 4℃, 5000rpm, and 1min, and the excess solution was scraped off with a cover glass, and the above-mentioned microneedle array was positioned and pressed into the large mold, and then it was dried at 37℃ for 6h and demolded to obtain the inflammation-responsive microneedle patch.

[0061] Example 3: Degradation experiment of inflammation-responsive microneedle patch for atopic dermatitis

[0062] The inflammatory responsive microneedle patch prepared in Example 1 was placed in 0.5 mL phosphate buffer (PBS) or 0.1 mg / mL collagenase solution. The surface moisture of the inflammatory responsive microneedle patch tip was dried at 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 48 h, 60 h, and 72 h. After weighing, 0.5 mL of fresh PBS or collagenase solution was replaced. Finally, the degradation rate was calculated by the weight change of each sample to determine the degradation curve of the microneedle patch, such as Figure 2 shown.

[0063] Example 4: Inflammation-responsive microneedle patch release experiment for atopic dermatitis

[0064] The inflammatory responsive microneedle patch prepared in Example 1 (containing 1 mg / mL rhodamine B, used to replace the therapeutic drug) was placed in 1 mL phosphate buffered saline (PBS) or 0.1 mg / mL collagenase solution. The release solution was aspirated at 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 36 h, 48 h, 60 h, and 72 h, and replaced with 1 mL of fresh PBS or collagenase solution. Finally, the drug release amount was calculated by the release solution of each sample at different time points to determine the release curve of the microneedle patch, such as Figure 3 As shown. This indicates that the release process of the inflammatory responsive microneedle patch for atopic dermatitis is related to the degradation process, and the inflammatory responsive release of the drug can be achieved through the degradation of matrix metalloproteinases, which is of great significance for alleviating the symptoms of atopic dermatitis.

[0065] The above are only preferred embodiments of the present invention, and the protection scope of the present invention is not limited to the above embodiments. All technical solutions under the concept of the present invention belong to the protection scope of the present invention. It should be pointed out that for ordinary technicians in this technical field, some improvements and modifications without departing from the principle of the present invention should be regarded as the protection scope of the present invention.

Claims

1. A method for preparing an inflammatory responsive microneedle patch for atopic dermatitis, characterized in that: The steps include: 1) Preparation of microneedle raw material solution: The raw material 1, the therapeutic drug and 1% 2-hydroxy-2-methylpropiophenone were uniformly mixed to obtain a microneedle raw material solution 1; the raw material 2 was dissolved in pure water, and the microneedle raw material solution 2 was obtained after centrifugation to remove bubbles, and stored at 4°C for later use; 2) Preparation of inflammation-responsive microneedle patches: The microneedle raw material solution 1 is added to the PDMS microneedle small mold, and the solution is uniformly filled into the needle body of the small mold by centrifugation or vacuuming, and then the excess solution is scraped off and cured by ultraviolet light, and the microneedle raw material solution 2 is added to serve as the inflammation-responsive microneedle patch substrate, and the microneedle array is obtained after drying and demolding; the microneedle raw material solution 2 is then added to the PDMS microneedle large mold, and the solution is uniformly filled into the needle body of the large mold by centrifugation or vacuuming, and then the excess solution is scraped off, and the above-mentioned microneedle array is positioned and pressed into the large mold, and then the microneedle array is dried and demolded to obtain the inflammation-responsive microneedle patch; In step (1), the microneedle raw material 1 is selected from methacrylate gelatin; the therapeutic drug is selected from triamcinolone acetonide, and the concentration is 20-25 mg / mL; In step (1), the microneedle raw material 2 is selected from sodium hyaluronate.

2. The preparation method according to claim 1, characterized in that: In step (2), in the two microneedle molds of different sizes, the needle tip length of the large microneedle mold is 0.4-1.5 mm, and the needle tip length of the small microneedle mold is 0.2-1 mm.

3. The preparation method according to claim 1, characterized in that: In step (2), in the two microneedle molds of different sizes, the bottom radius of the large microneedle mold is 100-600 μm, and the bottom radius of the small microneedle mold is 100-300 μm.

4. The preparation method according to claim 3, characterized in that: After adding 1% 2-hydroxy-2-methylpropiophenone to the microneedle raw material solution 1, the solution should be kept away from light.

5. The preparation method according to claim 3, characterized in that: The drying temperature is lower than 37°C.

6. An inflammatory responsive microneedle patch for atopic dermatitis, characterized in that: The preparation method is described in any one of claims 1 to 5.

Images