Micro-needle body, preparation method thereof, micro-needle application and application of micro-needle application

Abstract

The invention relates to the technical field of traditional Chinese medicine microneedle application, and discloses a microneedle body, a preparation method of the microneedle body, microneedle application and application of the microneedle application. The microneedle body comprises a needle tip, a separation layer and a substrate layer which are arranged in sequence, a material I for forming the needle tip comprises silver-loaded oxidized chitosan and a traditional Chinese medicine composition; a material II for forming the separation layer comprises a component A and magnesium particles; a material III for forming the substrate layer comprises silver-loaded oxidized chitosan; the traditional Chinese medicine composition contains an angelica sinensis extract, a prepared rehmannia root extract, a ligusticum wallichii extract, a radix paeoniae alba extract, peach kernel powder, a safflower extract, a rhizoma cyperi extract, a salvia miltiorrhiza extract and glycyrrhizic acid in a mass ratio of 1: (0.3-3): (0.3-3): (0.3-3): (0.2-3): (0.2-3): (0.2-3): (0.3-3): (0.3-2). The microneedle body has a good application prospect in preparation of an application for treating irregular menstruation caused by qi stagnation and blood stasis.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine microneedle application, and in particular to a microneedle body and a preparation method thereof, a microneedle application and an application thereof. Background Art

[0002] Menstrual irregularities refer to symptoms such as irregular menstrual cycles, periods, and menstrual blood volume in women of childbearing age. The causes are complex. For example, cold stimulation, emotional damage, improper diet, and many other factors can cause menstrual irregularities.

[0003] Qi stagnation and blood stasis syndrome is a common syndrome of irregular menstruation. Its cause is insufficient Qi and blood, and poor blood circulation, which leads to the formation of blood stasis. Therefore, Qi should be replenished, Qi and blood should be activated to make Qi and blood flow smoothly, which can naturally improve blood stasis. The key to curing the syndrome of irregular menstruation caused by Qi stagnation and blood stasis lies in strengthening the spleen and regulating Qi, replenishing Qi and blood, and activating blood circulation and removing blood stasis.

[0004] Currently, there are three main treatments for menstrual irregularities caused by Qi stagnation and blood stasis. The first is wearable heating patches, such as heat packs and uterine-warming belts. However, these apply heat physically, providing only temporary relief and not a cure. The second is the use of health-promoting foods, such as dysmenorrhea tea. However, these foods are slow to take effect, and some taste unpleasant. The third is the use of estrogen and progesterone to regulate the body's sex hormone balance and provide symptomatic treatment. While this method has good short-term efficacy, it is prone to recurrence after discontinuation, and long-term use of estrogen and progesterone can significantly impact the body's normal metabolism.

[0005] Studies have found that Fufang Siwu Decoction can significantly improve hormone levels, more effectively than conventional treatments. For example, Taohong Siwu Decoction can treat Qi stagnation and blood stasis syndrome by downregulating the expression of proteins associated with the Rho / ROCK pathway, providing a theoretical basis for the subsequent use of Taohong Siwu Decoction-related drugs in treating menstrual irregularities associated with Qi stagnation and blood stasis in clinical practice.

[0006] CN115252740A discloses a Chinese medicine composition for treating irregular menstruation. The composition, which fundamentally regulates the liver, kidneys, spleen, and stomach, and supplemented with blood-activating and tranquilizing drugs, restores the body to health while enhancing immunity and resistance, thereby achieving the purpose of regulating irregular menstruation. The composition has significant therapeutic effects on symptoms such as irregular menstrual cycles, scanty menstruation, blood stasis, dysmenorrhea, and amenorrhea. However, this prior art is less targeted at treating irregular menstruation due to qi stagnation and blood stasis.

[0007] Dissolvable microneedles are made of water-soluble materials and disperse drugs within the needle body, resulting in a high drug loading capacity, no needle tip waste after use, and lower production costs. Dissolvable, detachable microneedles, which remove the basal layer and allow the needle tip to remain in the skin with sustained drug release, have been widely studied by researchers.

[0008] CN110870846A discloses a rapidly implantable sustained-release microneedle, which has an intermediate layer between the needle tip and the base layer of a traditional microneedle, and the intermediate layer is composed of a matrix containing a hydrophilic material. After the microneedle is applied to the skin, the separation layer absorbs moisture, allowing the needle tip to quickly separate from the needle body and the base within 1 hour, so that the needle tip remains in the skin after the base is removed. Therefore, the user does not need to apply the microneedle patch for a long time to ensure the long-term release of the drug in the body. However, the needle tip separation time of the above-mentioned detachable microneedle patch is relatively long. When the microneedle patch is applied to a subject who is active and has limited endurance, the long separation time required may cause the microneedle patch to fall off or cause itching of the skin. Summary of the Invention

[0009] The purpose of the present invention is to overcome the problem of long needle tip separation time in the detachable microneedle patch in the prior art.

[0010] To achieve the above-mentioned object, the first aspect of the present invention provides a microneedle body, which comprises a needle tip, a separation layer and a base layer arranged in sequence; the length of the needle tip is 0.2-1.4 mm, the thickness of the separation layer is 0.1-0.6 mm, and the thickness of the base layer is 0.01-0.3 mm; The material I forming the needle tip includes silver-loaded oxidized chitosan and a traditional Chinese medicine composition; The material II forming the separation layer includes component A and magnesium particles; The material III forming the base layer includes silver-loaded oxidized chitosan; The traditional Chinese medicine composition contains angelica extract, rehmannia root extract, ligusticum chuanxiong extract, white peony root extract, peach kernel powder, safflower extract, cyperus rotundus extract, salvia miltiorrhiza extract and glycyrrhizic acid in a mass ratio of 1:0.3-3:0.3-3:0.3-3:0.2-3:0.2-3:0.2-3:0.3-3:0.3-2; Component A is a combination of sodium hyaluronate and polyvinyl pyrrolidone or a combination of sodium hyaluronate and sucrose; The average particle diameter of the magnesium particles is 10-100 μm.

[0011] The second aspect of the present invention provides a method for preparing the microneedle body according to the first aspect, the method comprising the following steps: (1) Pretreating material I containing silver-loaded oxidized chitosan and a traditional Chinese medicine composition in a microneedle concave mold to obtain a first intermediate product; (2) in the presence of alkaline solution I, subjecting the first intermediate product to a first gel treatment to obtain a needle tip; (3) performing a first drying treatment on the surface of the needle tip of the material II containing the component A and the magnesium particles to form a separation layer; (4) On the surface of the separation layer, the material III containing silver-loaded oxidized chitosan is subjected to a second drying treatment to form a base layer.

[0012] A third aspect of the present invention provides a microneedle patch, comprising a microneedle body and a dressing layer, each of which is independently stored; The dressing layer is a hydrogel formed by a material containing silver-loaded oxidized chitosan; The surface area of ​​the dressing layer is greater than or equal to the surface area of ​​the base layer of the microneedle body, so that the dressing layer completely covers the base layer of the microneedle body; The microneedle body is the microneedle body described in the first aspect.

[0013] The fourth aspect of the present invention provides use of the microneedle patch described in the third aspect in preparing a patch for treating menstrual irregularities caused by qi stagnation and blood stasis syndrome.

[0014] The present invention loads a traditional Chinese medicine composition into the needle tip, uses material II containing component A and magnesium particles of a specific size (with an average particle diameter of 10-100 μm) to form a separation layer, and uses material III containing silver-loaded oxidized chitosan to form a base layer. The obtained microneedle body has good application prospects in the preparation of a dressing for treating menstrual irregularities caused by qi stagnation and blood stasis.

[0015] The microneedle patch provided by the present invention has an extremely high transdermal drug delivery efficiency (85%-95%); and the microneedle patch overcomes the defect of a long needle tip separation time in traditional detachable microneedle patches. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 is a schematic diagram of the microneedle application F1 obtained in Application Example 1 of the present invention; Figure 2 This is a diagram showing the antibacterial effects of microneedle patches F1, F2, F3, and F5 obtained in Application Example 1 of the present invention. DETAILED DESCRIPTION

[0017] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0018] As described above, the first aspect of the present invention provides a microneedle body, which comprises a needle tip, a separation layer, and a base layer arranged in sequence; the length of the needle tip is 0.2-1.4 mm, the thickness of the separation layer is 0.1-0.6 mm, and the thickness of the base layer is 0.01-0.3 mm; The material I forming the needle tip includes silver-loaded oxidized chitosan and a traditional Chinese medicine composition; The material II forming the separation layer includes component A and magnesium particles; The material III forming the base layer includes silver-loaded oxidized chitosan; The traditional Chinese medicine composition contains angelica extract, rehmannia root extract, ligusticum chuanxiong extract, white peony root extract, peach kernel powder, safflower extract, cyperus rotundus extract, salvia miltiorrhiza extract and glycyrrhizic acid in a mass ratio of 1:0.3-3:0.3-3:0.3-3:0.2-3:0.2-3:0.2-3:0.3-3:0.3-2; Component A is a combination of sodium hyaluronate and polyvinyl pyrrolidone or a combination of sodium hyaluronate and sucrose; The average particle diameter of the magnesium particles is 10-100 μm.

[0019] Preferably, the needle tip is conical and / or polygonal-pyramid shaped.

[0020] Preferably, the polygonal pyramid is a quadrangular pyramid.

[0021] Preferably, the angle of the needle tip is 10-60°.

[0022] Preferably, relative to 1 cm 2 The base layer has a number of needle tips of 25-1000.

[0023] More preferably, relative to 1 cm 2 The base layer has 100-200 needle tips.

[0024] Preferably, the silver-loaded oxidized chitosan is prepared by a method comprising the following steps: S1: In the presence of a buffer solution I and oxygen, chitosan, tetramethylpiperidinyl oxide and laccase are subjected to a first heat treatment to obtain a mixture I; S2: subjecting the mixture I to a first crystallization treatment with anhydrous ethanol to obtain an intermediate I; S3: In the presence of buffer solution II, subjecting the intermediate I and silver nitrate to a second heating treatment to obtain intermediate II; S4: subjecting the intermediate II and acetone to a second crystallization treatment to obtain the silver-loaded oxidized chitosan.

[0025] Preferably, in step S1, the weight ratio of the chitosan to the tetramethylpiperidinyl oxide is 25-26:1.

[0026] Preferably, in step S1, the amount of the laccase used is 2500-2750 μL per 1 g of the tetramethylpiperidinyloxide.

[0027] Preferably, in step S1, the amount of the buffer solution I used is 2500-3000 mL for every 1 g of the tetramethylpiperidinyl oxide.

[0028] Preferably, in step S1, the conditions of the first heating treatment include: a temperature of 38-43°C and a time of 17-18 hours.

[0029] The present invention has no particular requirements for the first heat treatment method, and those skilled in the art can select a method as needed. For example, the first heat treatment is performed in a water bath.

[0030] The present invention has no particular requirements for the conditions of the first crystallization treatment and the second crystallization treatment, and those skilled in the art can select them as needed. For example, the conditions of the first crystallization treatment and the second crystallization treatment each independently include: a temperature of 15-25° C. and a time of 5-10 minutes.

[0031] According to a preferred embodiment, the method further comprises: in step S1, centrifuging and filtering the product after the first heat treatment in sequence to obtain the mixture I.

[0032] The present invention has no special requirements for the centrifugal conditions, and those skilled in the art can select them as needed.

[0033] The present invention has no particular requirements for the first filtration method, and those skilled in the art can select one as needed. For example, a 0.1 μm ultrafiltration membrane is used for the first filtration.

[0034] Preferably, the method further comprises: in step S2, subjecting the product after the first crystallization treatment to a second filtration and drying I in sequence to obtain the intermediate I.

[0035] Preferably, the amount of the anhydrous ethanol used is 50-85 mL for every 1 g of the tetramethylpiperidinyl oxide.

[0036] The present invention has no special requirements for the second filtering method, and those skilled in the art can select it as needed.

[0037] Preferably, in step S3, the conditions of the second heating treatment include: a temperature of 40-60° C. and a time of 1-2 hours.

[0038] Preferably, in step S3, the amount of the buffer solution II used is 125-150 mL for every 1 g of the intermediate I.

[0039] Preferably, in step S3, the amount of silver nitrate used is 1-1.2 mmol per 1 g of the intermediate I.

[0040] The present invention has no particular requirements for the state of the silver nitrate, and those skilled in the art can select it as needed. Preferably, a silver nitrate solution with a concentration of 5-15 mmol / L is used.

[0041] Preferably, the amount of acetone used is 2-10 mL per 1 g of the intermediate I.

[0042] According to a preferred embodiment, the method further comprises, in step S4, sequentially filtering, washing and drying II the product of the second crystallization treatment to obtain the silver-loaded oxidized chitosan.

[0043] The present invention has no particular requirements for the conditions of the filtration and washing, and those skilled in the art can select them as needed. For example, filtration is performed using a 0.01-0.1 μm filter membrane, and washing is performed 3-8 times with ethanol.

[0044] The present invention has no particular requirements for the conditions of drying I and drying II. It is only necessary to dry the product after the second filtration and the product after washing to constant weight. Those skilled in the art can select the conditions as needed. Exemplarily, vacuum drying is used.

[0045] Preferably, the buffer solution I and the buffer solution II are both provided by acetic acid-sodium acetate buffer solution with a pH value of 4.4-4.8.

[0046] Preferably, component A is a combination of sodium hyaluronate and polyvinyl pyrrolidone at a mass ratio of 1:40-60, or a combination of sodium hyaluronate and sucrose at a mass ratio of 1:30-40. The inventors of the present invention have discovered that in this preferred embodiment, the resulting microneedle body exhibits a shorter tip separation time for microneedle application.

[0047] Preferably, the polyvinyl pyrrolidone is selected from at least one of polyvinyl pyrrolidone K17PF, polyvinyl pyrrolidone K30, polyvinyl pyrrolidone K60 and polyvinyl pyrrolidone K90.

[0048] Preferably, the traditional Chinese medicine composition contains angelica extract, rehmannia root extract, ligusticum chuanxiong extract, white peony root extract, peach kernel powder, safflower extract, cyperus rotundus extract, salvia miltiorrhiza extract, and glycyrrhizic acid in a mass ratio of 1:0.5-1:0.5-1:1-2:0.3-1:0.3-1:0.3-1:1-2:0.5-1. The inventors of the present invention have discovered that, in this preferred embodiment, the resulting microneedle body exhibits a more effective effect in treating menstrual irregularities due to qi stagnation and blood stasis syndrome when used in microneedle application.

[0049] Preferably, the mass ratio of the silver-loaded oxidized chitosan to the traditional Chinese medicine composition in the material I is 1:2-4. The inventors of the present invention have found that in this preferred embodiment, the prepared microneedle body has better mechanical strength.

[0050] Preferably, the mass ratio of component A to magnesium particles in the material II is 1:0.05-0.4. The inventors of the present invention have found that in this preferred embodiment, the prepared microneedle body has a shorter needle tip separation time for microneedle application.

[0051] Preferably, the concentration of silver-loaded oxidized chitosan in the material III is 0.02-0.10 g / ml.

[0052] As mentioned above, the second aspect of the present invention provides a method for preparing the microneedle body of the first aspect, the method comprising the following steps: (1) Pretreating material I containing silver-loaded oxidized chitosan and a traditional Chinese medicine composition in a microneedle concave mold to obtain a first intermediate product; (2) in the presence of alkaline solution I, subjecting the first intermediate product to a first gel treatment to obtain a needle tip; (3) performing a first drying treatment on the surface of the needle tip of the material II containing the component A and the magnesium particles to form a separation layer; (4) On the surface of the separation layer, the material III containing silver-loaded oxidized chitosan is subjected to a second drying treatment to form a base layer.

[0053] According to a preferred embodiment, the pretreatment operation includes: firstly subjecting the material I containing silver-loaded oxidized chitosan and the traditional Chinese medicine composition to a first centrifugation in a microneedle concave mold to obtain the first intermediate product.

[0054] The present invention has no particular requirements for the conditions of the first centrifugation, and those skilled in the art can select them as needed. For example, the first centrifugation is performed at a temperature of 20-25° C. and a rotation speed of 1000-8000 rpm for 5-15 minutes.

[0055] Preferably, in step (2), the alkaline solution I is provided by a sodium hydroxide solution with a concentration of 8-15 g / mL.

[0056] Preferably, the amount of the alkaline solution I is controlled so that the first intermediate product can be completely gelled.

[0057] Preferably, in step (2), the conditions for the first gel treatment include: a temperature of 40-80° C. and a time of 2-4 hours.

[0058] Preferably, the method further comprises: in step (2), subjecting the product after the first gel treatment to a first washing and drying treatment I in sequence to obtain a needle tip.

[0059] The present invention has no particular requirements for the conditions of the first washing and drying treatments I, and those skilled in the art can select them as needed. For example, the first washing is performed with flowing distilled water for 1-3 minutes, followed by drying treatment I at 50-70°C for 3-8 minutes.

[0060] Preferably, in step (3), the conditions of the first drying treatment include: a temperature of 20-60°C and a time of 2-24 hours.

[0061] Preferably, the method further comprises: in step (3), first subjecting the material II containing component A and magnesium particles to a second centrifugation, and then subjecting the product of the second centrifugation to a first drying treatment to form a separation layer.

[0062] The present invention has no particular requirements for the conditions of the second centrifugation, and those skilled in the art can select them as needed. For example, the second centrifugation is performed at a temperature of 20-25° C. and a rotation speed of 500-1000 rpm for 5-15 minutes.

[0063] Preferably, in step (4), the material III containing silver-loaded oxidized chitosan is prepared by a method comprising the following steps: The organic solution I containing silver-loaded oxidized chitosan was dialyzed, and then the pH value was adjusted to 5-7, and rotary evaporated at 35-50° C. for 10-30 min to obtain material III containing silver-loaded oxidized chitosan.

[0064] Preferably, the dialysis treatment conditions include: a temperature of 18-28° C. and a time of 36-48 h.

[0065] Preferably, both the material I and the material II are used in the form of a solution and / or a dispersion.

[0066] Preferably, the material I is prepared by a method comprising the following steps: Dissolving each component of the traditional Chinese medicine composition in polyethylene glycol to obtain a traditional Chinese medicine mixture containing the traditional Chinese medicine composition; then mixing material III containing silver-loaded oxidized chitosan and the traditional Chinese medicine mixture containing the traditional Chinese medicine composition to obtain material I; The material III is the same as the material III in step (4); The Chinese medicine composition is the same as the Chinese medicine composition described in the first aspect.

[0067] Preferably, the pH value of the traditional Chinese medicine mixture is 5.8-6.2.

[0068] The present invention has no particular requirements for the mixing conditions, and those skilled in the art can select them as needed. For example, the mixing is performed at 20-35° C. for 5-10 minutes.

[0069] Preferably, the solvent in the material II is selected from at least one of water, ethanol and acetic acid.

[0070] The present invention has no particular requirements on the type of water, and those skilled in the art can select it as needed. For example, it can be deionized water and / or ultrapure water.

[0071] Preferably, the concentration of component A in the material II is 5-40 g / mL.

[0072] Preferably, the second drying step comprises: firstly performing a first stage drying at 20-25° C. for 5-15 minutes, and then performing a second stage drying at 50-70° C. for 4-6 hours to form a base layer.

[0073] As mentioned above, the third aspect of the present invention provides a microneedle patch, which includes a microneedle body and a dressing layer that are each stored independently; The dressing layer is a hydrogel formed by a material containing silver-loaded oxidized chitosan; The surface area of ​​the dressing layer is greater than or equal to the surface area of ​​the base layer of the microneedle body, so that the dressing layer completely covers the base layer of the microneedle body; The microneedle body is the microneedle body described in the first aspect.

[0074] Preferably, the dressing layer is prepared by a method comprising the following steps: In the presence of alkaline solution II, the organic solution II containing silver-loaded oxidized chitosan is subjected to a second gel treatment to obtain the dressing layer.

[0075] Preferably, the thickness of the dressing layer is 3-6 mm.

[0076] Preferably, the second gel treatment is performed in a dressing layer mold of 10 mm×10 mm×3 mm to 20 mm×20 mm×6 mm.

[0077] Preferably, the alkaline solution II is provided by a sodium hydroxide solution with a concentration of 5-20 g / mL.

[0078] Preferably, the conditions for the second gel treatment include: a temperature of 15-25° C. and a time of 20-30 min.

[0079] Preferably, the content of silver-loaded oxidized chitosan in the organic solution II is 0.01-0.2 g / ml.

[0080] Preferably, the amount of the alkaline solution II is controlled so that the silver-loaded oxidized chitosan in the organic solution II can be completely gelled.

[0081] In the present invention, the silver-loaded oxidized chitosan in the dressing layer is the same as the silver-loaded oxidized chitosan described in the first aspect.

[0082] In the present invention, the solvents in the organic solution I and the organic solution II are each independently a mixed solution of acetic acid and water in a volume ratio of 1:2-5.

[0083] As mentioned above, the fourth aspect of the present invention provides the use of the microneedle patch described in the third aspect in the preparation of a patch for treating menstrual irregularities caused by qi stagnation and blood stasis syndrome.

[0084] The present invention will be described in detail below through examples. In the following examples, unless otherwise specified, the instruments, reagents, materials, etc. involved are all conventional instruments, reagents, materials, etc., which can be obtained through regular commercial channels. In particular, unless otherwise specified, the reagents used are all commercially available analytical grade products.

[0085] In the following examples and preparation examples, the buffer solution I and the buffer solution II are both provided by an acetic acid-sodium acetate buffer solution with a pH value of 4.6.

[0086] In the following examples and preparation examples, alkaline solution I and alkaline solution II are both provided by sodium hydroxide solution with a concentration of 10 g / mL.

[0087] Both organic solution I and organic solution II are mixed solutions consisting of acetic acid and water in a volume ratio of 1:3; Chitosan: purchased from Shanghai MacLean Biochemical Technology Co., Ltd., brand C766421.

[0088] Tetramethylpiperidinyl oxide: purchased from Nanjing Huikang Biotechnology Co., Ltd., CAS number is 2564-83-2.

[0089] Laccase: purchased from Shanghai Yuanye Biotechnology Co., Ltd., brand S10189.

[0090] Magnesium particles: Magnesium particles I: The average particle diameter is 20 μm, purchased from Zhejiang Yamei Nano Technology Co., Ltd. and customized.

[0091] Magnesium particles II: The average particle diameter is 200 μm, purchased from Xinxiang County Jinghua Magnesium Industry Co., Ltd. and customized.

[0092] Polyvinylpyrrolidone: Polyvinylpyrrolidone K90: PVPK90 was purchased from Shanghai MacLean Biochemical Technology Co., Ltd. with the brand name P823856.

[0093] Polyvinylpyrrolidone K30: PVPK30 was purchased from Shanghai MacLean Biochemical Technology Co., Ltd. with the brand name P816205.

[0094] Sucrose: purchased from Changde Beekman Biotechnology Co., Ltd.

[0095] Sodium hyaluronate: purchased from Shanghai Yuanye Biotechnology Co., Ltd., brand number S12034.

[0096] Angelica extract: purchased from Shaanxi Tianxingjian Biochemical Technology Co., Ltd.

[0097] Rehmannia root extract: purchased from Shaanxi Tianxingjian Biochemical Technology Co., Ltd. Chuanxiong extract: purchased from Shaanxi Tianxingjian Biochemical Technology Co., Ltd.

[0098] White peony root extract: purchased from Shaanxi Tianxingjian Biochemical Technology Co., Ltd.

[0099] Peach kernel powder: purchased from Shaanxi Tianxingjian Biochemical Technology Co., Ltd.

[0100] Safflower extract: purchased from Shaanxi Tianxingjian Biochemical Technology Co., Ltd.

[0101] Cyperus rotundus extract: purchased from Shaanxi Tianxingjian Biochemical Technology Co., Ltd.

[0102] Salvia miltiorrhiza extract: purchased from Shaanxi Tianxingjian Biochemical Technology Co., Ltd.

[0103] Glycyrrhizic acid: purchased from Shaanxi Tianxingjian Biochemical Technology Co., Ltd., CAS No. 1405-86-3.

[0104] Microneedle mold: 144 needles / cm 2 , purchased from Anhui Zhongding Yuxuan New Materials Technology Co., Ltd., model MN-H400.

[0105] Dressing layer mold: 20mm×20mm×6mm.

[0106] Preparation Example A: Preparation of silver-loaded oxidized chitosan S1: In the presence of 300 mL of buffer solution I and oxygen, 3 g of chitosan, 0.12 g of tetramethylpiperidinyl oxide, and 300 μL of laccase were subjected to a first heat treatment in a 40°C water bath for 18 h, followed by centrifugation and a first filtration using a 0.1 μm ultrafiltration membrane to obtain a mixture I; S2: The mixture I and 10 mL of anhydrous ethanol were subjected to a first crystallization treatment at 20° C. for 5 min, followed by a second filtration and vacuum drying to obtain an intermediate I; S3: In the presence of 45 mL of buffer solution II, 0.36 g of the intermediate I and 36 mL of silver nitrate solution (concentration of 10 mmol / L) were subjected to a second heating treatment at 50° C. for 1.5 h to obtain intermediate II; S4: The intermediate II and 2 mL of acetone were subjected to a second crystallization treatment at 25° C. for 5 min. The product of the second crystallization treatment was filtered using a 0.01 μm filter membrane, washed with ethanol 5 times, and then vacuum-dried to obtain silver-loaded oxidized chitosan.

[0107] The following preparation examples of series B are used to illustrate the preparation of the dressing layer.

[0108] Preparation Example B1 In the dressing layer mold, in the presence of 30 mL of alkaline solution II, 10 mL of organic solution II containing 0.2 g of silver-loaded oxidized chitosan was subjected to a second gelation treatment at 20° C. for 30 min to obtain dressing layer N1.

[0109] Preparation Example B2 This preparation example was carried out in a similar manner to that of Preparation Example B1, except that an equal mass of chitosan was used to replace the silver-loaded oxidized chitosan in Preparation Example B1. The remaining steps are the same as those in Preparation Example B1 to obtain dressing layer N2.

[0110] Preparation Example C This preparation example is used to illustrate the preparation of Material III containing silver-loaded oxidized chitosan by a method comprising the following steps.

[0111] 4 g of silver-loaded oxidized chitosan was dissolved in 100 mL of organic solution I to obtain organic solution I containing silver-loaded oxidized chitosan; the organic solution I containing silver-loaded oxidized chitosan was dialyzed at 25° C. for 36 h, the pH value was adjusted to 6, and then rotary evaporated at 35° C. for 20 min to obtain material III; The concentration of silver-loaded oxidized chitosan in Material III was determined to be 0.047 g / mL by high performance liquid chromatography (HPLC-ELSD).

[0112] The following preparation examples of series D are used to illustrate the preparation of material I containing silver-loaded oxidized chitosan and a traditional Chinese medicine composition with reference to the formula in Table 1.

[0113] Preparation Example D1 The components of the traditional Chinese medicine composition were dissolved in polyethylene glycol to obtain a traditional Chinese medicine mixture I containing the traditional Chinese medicine composition; then, Material III containing silver-loaded oxidized chitosan (obtained in the above Preparation Example C) and the traditional Chinese medicine mixture I containing the traditional Chinese medicine composition were mixed at 25° C. for 10 minutes to obtain Material I, which was named Material I-1.

[0114] Unless otherwise specified, the preparation examples of the remaining D series were prepared using a method similar to that of Preparation Example D1, except that the formula of the Chinese medicine mixture was different, as shown in Table 1. The unlisted parts were the same as those of Preparation Example D1.

[0115] Table 1

[0116] Preparation Example E This preparation example is used to illustrate the preparation of Material II containing component A and magnesium particles according to the formulation in Table 2 by a method comprising the following steps: Component A and magnesium particles are dispersed in a solvent to obtain material II.

[0117] Table 2

[0118] Example 1 This example is used to illustrate the preparation of a microneedle body by referring to the formula and process parameters in Table 3 and by a method comprising the following steps.

[0119] (1) In a microneedle concave mold, material I containing silver-loaded oxidized chitosan and a traditional Chinese medicine composition is first centrifuged at 25° C. and 2000 rpm for 10 min to obtain a first intermediate product; (2) In the presence of 10 mL of alkaline solution I, the first intermediate product is subjected to a first gel treatment, and the product after the first gel treatment is washed with flowing distilled water for 2 minutes, and dried at 60° C. for 15 minutes to obtain a needle tip; (3) On the surface of the needle tip, firstly subjecting the material II containing the component A and the magnesium particles to a second centrifugation at 25° C. and a rotation speed of 500 rpm for 10 min, and then subjecting the product of the second centrifugation to a first drying treatment to form a separation layer; (4) On the surface of the separation layer, the material III containing silver-loaded oxidized chitosan was first dried at 25°C for 10 minutes and then dried at 60°C for 5 hours in the second stage to form a base layer.

[0120] Unless otherwise specified, the remaining examples were carried out using the same process as Example 1, except that the raw material formulations and process parameters were different, as shown in Table 3. The unlisted parts were the same as Example 1.

[0121] Table 3

[0122] Example 4 This example is carried out using a method similar to that of Example 1, except that an equal volume of material I-4 is used to replace material I-1 in Example 1. The remaining steps are the same as those of Example 1 to obtain a microneedle body named H4.

[0123] Comparative Example 1 This example is carried out using a method similar to that of Example 1, except that this example uses material II-D1 of equal mass to replace material II-1 in Example 1, and the remaining steps are the same as those of Example 1 to obtain a microneedle body named D-H1.

[0124] Comparative Example 2 This comparative example was carried out using a method similar to that of Example 1, except that an equal volume of material II-D2 was used to replace material II-1 in Example 1. The remaining steps were the same as those of Example 1 to obtain a microneedle body named D-H2.

[0125] Application Example 1 During the application process, the dressing layer prepared in the above example and the microneedle body were combined to obtain a microneedle patch. The obtained microneedle patch is shown in Table 4.

[0126] Table 4

[0127] Test example: 1. Analyze transdermal drug delivery efficiency Four pieces of fresh, clean, depilated pigskin tissue of equal area (20 mm × 30 mm) were obtained and numbered I, II, III, and IV. Topical medications were then stained. Pigskin No. I was smeared with 20 ml of the Chinese herbal mixture I (obtained in Preparation Example D1 above); pigskin No. II was injected with 20 ml of the Chinese herbal mixture I; pigskin No. III was injected with a transdermal patch; and pigskin No. IV was implanted with the microneedle applicator F1 described in the above example. After 20 minutes, the drug-stained areas were observed under a microscope for the different drug administration methods. The drug penetration and absorption rates were expressed using the stained areas, and the drug administration efficiency was calculated. The results are shown in Table 5.

[0128] Drug delivery efficiency = infected area / pigskin tissue area × 100%; Table 5

[0129] The preparation method of the transdermal patch in Table 5 is as follows: 40 g of polyisobutylene hot melt adhesive was melted, 3 g of azone, 5 g of glycerol, and 20 g of the Chinese medicine mixture I (obtained in the above Preparation Example D1) were mixed in a screw extruder, extruded, and coated onto a polyester film patch substrate, which was then rolled up to form a transdermal patch.

[0130] It can be seen from Table 5 that: when using the smear and transdermal patch delivery methods, only 10%-20% and 20-30% of the drug respectively penetrate the skin, which will result in most of the drug being wasted; when using the injection needle delivery method, 90%-100% of the drug penetrates the skin, but the injection needle penetrates into the dermis where pain receptors exist, which is very painful and leads to poor patient compliance; and when using the microneedle application delivery method, 85%-95% of the drug enters the skin, and this delivery method is painless. Combined with the principle analysis: the main reason why microneedle application has such a high drug delivery efficiency is that microneedle application can bypass the stratum corneum barrier and deliver the drug directly to the epidermis or upper dermis.

[0131] 2. The microneedle patches F1-F5, D-F1, and D-F2 obtained in the above examples were applied to fresh, cleaned pig skin tissues, and the peeling time between the needle tip and the basal layer was recorded. The results are shown in Table 6.

[0132] Table 6

[0133] It can be seen from the data in Table 6 that the glass time between the needle tip and the base layer in the microneedle patch provided by the present invention is only 4-5 minutes, which overcomes the defect of long needle tip separation time in traditional detachable microneedle patches.

[0134] 3. The antibacterial properties of the microneedle patches F1-F3 and F5 obtained in the above examples were studied using the following method. The specific method includes: (i) Microneedle patches F1-F3 and F5 were diluted with anhydrous methanol so that the concentration of chitosan or silver-loaded oxidized chitosan in the diluted solutions was 2 mg / mL; anhydrous methanol was used as a blank control group; (ii) On a clean bench, when the LB medium poured into the plate solidifies, add the prepared 10 6 100 μL of bacterial suspension of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and live Salmonella typhimurium at FU / mL was injected into a round plate and spread evenly; (iii) The diluted solution from step (i) was evenly applied onto a sterilized circular filter paper disc (0.3 cm in diameter), fixed onto a plate coated with the test bacteria, and cultured in a 37°C incubator for 24 h. (iv) Take the contact surface culture and make a serial dilution (10-fold dilution until the dilution is suitable for use), take 1 mL of the diluted bacterial suspension on the plate, pour agar into the plate, mix well, wait for it to solidify, invert and incubate at 37℃ for 24 hours, calculate the inhibition rate, and the results are as follows: Figure 2 shown.

[0135] Inhibition rate (%) = (total number of colonies in blank control - total number of colonies in gel) / total number of colonies in blank control × 100%.

[0136] pass Figure 2 It can be seen that the antibacterial effects of the microneedle application F1-F3 provided by the present invention on the four bacteria can reach more than 95%, and the antibacterial effects of the microneedle application F5 on various bacteria are all above 80%. Silver-loaded oxidized chitosan is added to the hydrogel dressing of the embodiment, which increases the loading capacity of the hydrogel dressing for the active ingredients, thereby achieving a higher antibacterial effect.

[0137] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, various simple variations of the technical solution of the present invention may be made, including combining the various technical features in any other appropriate manner. These simple variations and combinations should also be regarded as disclosed in the present invention and fall within the scope of protection of the present invention.

Claims

1. A microneedle body, characterized in that: The microneedle body comprises a needle tip, a separation layer and a base layer arranged in sequence; the length of the needle tip is 0.2-1.4 mm, the thickness of the separation layer is 0.1-0.6 mm, and the thickness of the base layer is 0.01-0.3 mm; The material I forming the needle tip includes silver-loaded oxidized chitosan and a traditional Chinese medicine composition; The material II forming the separation layer includes component A and magnesium particles; The material III forming the base layer includes silver-loaded oxidized chitosan; The traditional Chinese medicine composition contains angelica extract, rehmannia root extract, ligusticum chuanxiong extract, white peony root extract, peach kernel powder, safflower extract, cyperus rotundus extract, salvia miltiorrhiza extract and glycyrrhizic acid in a mass ratio of 1:0.3-3:0.3-3:0.3-3:0.2-3:0.2-3:0.2-3:0.3-3:0.3-2; Component A is a combination of sodium hyaluronate and polyvinyl pyrrolidone or a combination of sodium hyaluronate and sucrose; The average particle diameter of the magnesium particles is 10-100 μm.

2. The microneedle body according to claim 1, wherein: The angle of the needle tip is 10-60°; and / or, relative to 1 cm 2 The base layer has a number of needle tips of 25-1000.

3. The microneedle body according to claim 1, wherein: The silver-loaded oxidized chitosan is prepared by a method comprising the following steps: S1: In the presence of a buffer solution I and oxygen, chitosan, tetramethylpiperidinyl oxide and laccase are subjected to a first heat treatment to obtain a mixture I; S2: subjecting the mixture I to a first crystallization treatment with anhydrous ethanol to obtain an intermediate I; S3: In the presence of buffer solution II, subjecting the intermediate I and silver nitrate to a second heating treatment to obtain intermediate II; S4: subjecting the intermediate II and acetone to a second crystallization treatment to obtain the silver-loaded oxidized chitosan.

4. The microneedle body according to any one of claims 1 to 3, wherein: Component A is a combination of sodium hyaluronate and polyvinyl pyrrolidone in a mass ratio of 1:40-60, or component A is a combination of sodium hyaluronate and sucrose in a mass ratio of 1:30-40; And / or, the traditional Chinese medicine composition contains angelica extract, rehmannia root extract, ligusticum chuanxiong extract, white peony root extract, peach kernel powder, safflower extract, cyperus rotundus extract, salvia miltiorrhiza extract and glycyrrhizic acid in a mass ratio of 1:0.5-1:0.5-1:1-2:0.3-1:0.3-1:0.3-1:1-2:0.5-1.

5. The microneedle body according to any one of claims 1 to 4, wherein: The mass ratio of the silver-loaded oxidized chitosan and the traditional Chinese medicine composition in the material I is 1:2-4; And / or, the mass ratio of component A to magnesium particles in the material II is 1:0.05-0.4; And / or, the concentration of silver-loaded oxidized chitosan in the material III is 0.02-0.10 g / ml.

6. A method for preparing the microneedle body according to any one of claims 1 to 5, characterized in that: The method comprises the following steps: (1) Pretreating material I containing silver-loaded oxidized chitosan and a traditional Chinese medicine composition in a microneedle concave mold to obtain a first intermediate product; (2) in the presence of alkaline solution I, subjecting the first intermediate product to a first gel treatment to obtain a needle tip; (3) performing a first drying treatment on the surface of the needle tip of the material II containing the component A and the magnesium particles to form a separation layer; (4) On the surface of the separation layer, the material III containing silver-loaded oxidized chitosan is subjected to a second drying treatment to form a base layer.

7. The method according to claim 6, wherein: In step (4), the material III containing silver-loaded oxidized chitosan is prepared by a method comprising the following steps: The organic solution I containing silver-loaded oxidized chitosan was dialyzed, and then the pH value was adjusted to 5-7, and rotary evaporated at 35-50° C. for 10-30 min to obtain material III containing silver-loaded oxidized chitosan.

8. A microneedle patch, characterized in that: The microneedle patch includes a microneedle body and a dressing layer that are stored independently; The dressing layer is a hydrogel formed by a material containing silver-loaded oxidized chitosan; The surface area of ​​the dressing layer is greater than or equal to the surface area of ​​the base layer of the microneedle body, so that the dressing layer completely covers the base layer of the microneedle body; The microneedle body is the microneedle body according to any one of claims 1 to 5.

9. The microneedle patch according to claim 8, wherein The dressing layer is prepared by a method comprising the following steps: In the presence of alkaline solution II, the organic solution II containing silver-loaded oxidized chitosan is subjected to a second gel treatment to obtain the dressing layer.

10. Use of the microneedle patch according to claim 8 or 9 in the preparation of a patch for treating menstrual irregularities due to qi stagnation and blood stasis syndrome.

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