Tanshinone IIA double-layer polymer soluble acupoint microneedle capable of reducing fat and reducing weight as well as preparation method and application thereof

Abstract

The invention discloses a lipid-lowering weight-losing tanshinone IIA double-layer polymer soluble acupoint microneedle and a preparation method and application thereof.The microneedle is of a layered structure design, and a needle body of the microneedle is prepared from a tanshinone IIA ethanol-water solution, polyvinylpyrrolidone K30 and chondroitin sulfate; the substrate of the backing layer is prepared from a tanshinone IIA ethanol-water solution and polyvinyl alcohol. The insoluble drug tanshinone IIA is successfully loaded in the soluble microneedle through a specific ethanol-water co-solvent system and a micro-molding forming process, the microneedle array is complete in form, sharp in needle shape and excellent in mechanical strength, and can effectively puncture the skin cuticle, after puncturing, the needle body can be rapidly dissolved in the skin interstitial fluid, and therefore the skin cuticle can be effectively punctured. The efficient release of the medicine is realized; the irritation to the skin is avoided. The core innovation of the invention lies in that the modern microneedle transdermal drug delivery technology and the traditional Chinese medicine acupoint therapy are combined for application, so that the dyslipidemia condition caused by hyperlipidemia diet can be obviously regulated, and the effects of regulating body weight and reducing blood fat can be achieved.

Description

Technical Field

[0001] This invention relates to the field of biomedical materials technology, specifically to a tanshinone IIA bilayer polymer soluble acupoint microneedle that can reduce lipids and weight, its preparation method, and its application. Background Technology

[0002] Tanshinone IIA (TSA) is an important active ingredient in the treatment of cardiovascular and cerebrovascular diseases, but its extremely poor water solubility severely limits its oral bioavailability, while injection administration has problems such as being invasive and having poor compliance.

[0003] While dissolving microneedles (DMNs) offer a novel approach to non-invasive transdermal drug delivery, their hydrophilic matrix presents a fundamental contradiction with the strong hydrophobicity of tanshinone IIA. Under current manufacturing processes, applying existing microneedle technology to tanshinone IIA leads to difficulties in drug loading and crystallization due to the incompatibility between the strong hydrophobicity of tanshinone IIA and the hydrophilic matrix of the microneedles. This not only reduces drug loading and release performance but also severely weakens the mechanical strength and puncture resistance of the microneedles, making effective skin penetration difficult. Secondly, drug crystallization and the potential use of irritating excipients pose risks of erythema and edema after microneedle application, making skin irritation a safety concern, and the in vivo solubility is also uncontrollable. Furthermore, most current research focuses on conventional transdermal drug delivery and fails to integrate microneedle technology with traditional Chinese medicine acupoint theory, thus failing to verify and leverage the superior efficacy of acupoint-based drug delivery. Therefore, developing a tanshinone IIA soluble microneedle (TSA-DMNs) that can systematically solve the above problems, ensure non-irritation, have sufficient drug loading and excellent mechanical strength, achieve good in vivo dissolution and drug release performance, and ultimately verify its superior efficacy for acupoint drug delivery has become a technical bottleneck that urgently needs to be overcome in this field. Summary of the Invention

[0004] To address the above shortcomings, this invention provides a soluble microneedle made of tanshinone IIA bilayer polymer that can reduce lipids and weight, along with its preparation method and application. This solves the safety hazards caused by the poor water solubility of tanshinone IIA, low drug loading and release capacity of the microneedle, weak mechanical strength and puncture performance, and strong skin irritation.

[0005] To achieve the above objectives, the technical solution of the present invention is as follows: A method for preparing a tanshinone IIA bilayer polymer soluble acupoint microneedle that can reduce lipids and promote weight loss, comprising a needle matrix and a backing matrix. The needle matrix is ​​prepared from 1-10 parts of tanshinone IIA, 1000-5000 parts of polyvinylpyrrolidone K30 and 1000-5000 parts of chondroitin sulfate. The backing matrix is ​​prepared from 20-75 parts of tanshinone IIA and 5000-15000 parts of polyvinyl alcohol.

[0006] The aforementioned needle matrix is ​​prepared from 5-10 parts of tanshinone IIA, 1000-3000 parts of polyvinylpyrrolidone K30 and 1000-3000 parts of chondroitin sulfate, and the backing matrix is ​​prepared from 50-75 parts of tanshinone IIA and 8000-15000 parts of polyvinyl alcohol.

[0007] Specifically, the aforementioned needle matrix is ​​prepared from 10 parts of tanshinone IIA, 1000 parts of polyvinylpyrrolidone K30, and 1000 parts of chondroitin sulfate, and the backing matrix is ​​prepared from 75 parts of tanshinone IIA and 15000 parts of polyvinyl alcohol.

[0008] The aforementioned tanshinone IIA is first dissolved in a 20-70% ethanol-water solution before the preparation of the needle matrix and backing matrix. The concentration of tanshinone IIA in the ethanol-water solution is 1-7 mg / mL.

[0009] Specifically, the aforementioned tanshinone IIA is first dissolved in a 60% ethanol-water solution before the preparation of the needle matrix and backing matrix. The concentration of tanshinone IIA in the ethanol-water solution is 5 mg / mL.

[0010] The aforementioned method for preparing tanshinone IIA bilayer polymer-soluble acupoint microneedles includes a needle body preparation method and a backing layer preparation method, comprising the following steps: (1) Needle preparation method: Weigh 10mg of tanshinone IIA and dissolve it fully in 2mL of 20-70% ethanol-water solution to prepare 1-10mg / mL tanshinone IIA ethanol-water solution. Weigh 1g of polyvinylpyrrolidone K30 and 1g of chondroitin sulfate and dissolve them in tanshinone IIA ethanol-water solution to prepare the needle matrix. Pour the matrix into a soluble acupoint microneedle mold, centrifuge at 3000-5000r / min for 5-15min, and dry at room temperature for 5-15min to obtain the needle body. The needle length is 350µm-650µm, the needle array is 137, the thickness is 8mm, the outer diameter is 26mm, and the groove depth is 2.5mm. (2) Preparation method of backing layer: Weigh 75mg tanshinone IIA and dissolve it fully in 15mL of 20-70% ethanol-water solution to prepare 1-10mg / mL tanshinone IIA ethanol-water solution. Weigh 1.5g polyvinyl alcohol and dissolve it in tanshinone IIA ethanol-water solution to prepare backing matrix. Inject it into a mold containing the needle body in step (1). Centrifuge at 3000-5000r / min for 5-15min. Dry at room temperature for 20-30h. After demolding, tanshinone IIA soluble acupoint microneedles are obtained.

[0011] Specifically, the preparation method of the aforementioned tanshinone IIA bilayer polymer soluble acupoint microneedles includes a needle body preparation method and a backing layer preparation method, comprising the following steps: (1) Needle preparation method: Weigh 10mg of tanshinone IIA and dissolve it fully in 2mL of 60% ethanol-water solution to prepare 5mg / mL tanshinone IIA ethanol-water solution. Weigh 1g of polyvinylpyrrolidone K30 and 1g of chondroitin sulfate and dissolve them in tanshinone IIA ethanol-water solution to prepare needle matrix. Pour the matrix into a soluble acupoint microneedle mold, centrifuge at 4000r / min for 10min and dry at room temperature for 10min to obtain the needle body. The needle body has a needle length of 550µm, a needle array of 137 needles, a thickness of 8mm, an outer diameter of 26mm, and a groove depth of 2.5mm. (2) Preparation method of backing layer: Weigh 75mg tanshinone IIA and dissolve it fully in 15mL 60% ethanol-water solution to prepare 5mg / mL tanshinone IIA ethanol-water solution. Weigh 1.5g polyvinyl alcohol and dissolve it in tanshinone IIA ethanol-water solution to prepare backing matrix. Inject it into a mold containing needles, centrifuge at 4000r / min for 10min, dry at room temperature for 24h, and after demolding, tanshinone IIA soluble acupoint microneedles are obtained.

[0012] The aforementioned application of tanshinone IIA bilayer polymer soluble acupoint microneedles in drug formulations for improving hyperlipidemia.

[0013] The beneficial effects of this invention are: (1) The tanshinone IIA soluble acupoint microneedles prepared according to the preparation method of the present invention have good morphological characterization and good mechanical strength and puncture performance.

[0014] (2) The 550µm tanshinone IIA soluble acupoint microneedles screened according to the preparation method of the present invention have higher drug release performance.

[0015] (3) The relative equilibrium time of drug release of the tanshinone IIA soluble acupoint microneedles prepared according to the preparation method of the present invention is about 48 hours in both the Tanzhong acupoint and non-meridian non-acupoint skin. Among them, the permeability of the skin in the Tanzhong acupoint area is better than that in non-meridian non-acupoint areas.

[0016] (4) The tanshinone IIA soluble acupoint microneedles prepared according to the preparation method of the present invention can be quickly and completely dissolved after being inserted into the skin and coming into contact with the interstitial fluid of the skin, and have no irritation to mouse skin.

[0017] (5) The tanshinone IIA soluble acupoint microneedles of the present invention can significantly improve blood lipid abnormalities caused by high-fat diet, and can significantly reduce weight and regulate blood lipids. Attached Figure Description

[0018] Figure 1 Technology roadmap; Figure 2 Schematic diagram of a 550µm DMNs mold; Figure 3 : A diagram of acupoints on a mouse; Figure 4 Morphological examination of TSA-DMNs; Figure 5 Mechanical strength of TSA-DMNs at different needle lengths (A: mechanical strength of TSA-DMNs with a needle length of 350µm; B: mechanical strength of TSA-DMNs with a needle length of 450µm; C: mechanical strength of TSA-DMNs with a needle length of 550µm; D: mechanical strength of TSA-DMNs with a needle length of 650µm). Figure 6 : Puncture performance of TSA-DMNs with different needle lengths (A is the puncture performance of TSA-DMNs with a needle length of 350µm; B is the puncture performance of TSA-DMNs with a needle length of 450µm; C is the puncture performance of TSA-DMNs with a needle length of 550µm). Figure 7 Chromatograms of TSA reference standard, TSA-DMNs sample, and blank-DMNs sample (A is the chromatogram of TSA reference standard; B is the chromatogram of TSA-DMNs sample; C is the chromatogram of blank-DMNs sample). Figure 8 : Standard curve diagram of TSA reference solution; Figure 9 Comparison of drug release performance of TSA-DMNs with different needle lengths; Figure 10 : The morphology and solubility of 550µm TSA-DMNs at 0, 1, 10, 15, 20, and 25 min (A = 0 min; B = 1 min; C = 10 min; D = 15 min; E = 20 min; F = 25 min). Figure 11 : In vitro drug release performance of 550µm TSA-DMNs in acupoint and non-acupoint areas; Figure 12: Schematic diagram of DMNs intervention (A is the acupoint microneedle group; B is the blank microneedle group). Detailed Implementation

[0019] To enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention will be further described below with reference to specific embodiments, but the embodiments are not intended to limit the present invention.

[0020] Example 1: Preparation of soluble acupoint microneedles made from tanshinone IIA bilayer polymer that can lower lipids and reduce weight. The tanshinone IIA bilayer polymer soluble acupoint microneedle, which can reduce lipids and promote weight loss, consists of a needle matrix and a backing matrix. The needle matrix is ​​prepared from 10 parts of tanshinone IIA, 1000 parts of polyvinylpyrrolidone K30 and 1000 parts of chondroitin sulfate, while the backing matrix is ​​prepared from 75 parts of tanshinone IIA and 15000 parts of polyvinyl alcohol.

[0021] The mass ratio of the needle matrix and backing matrix described above was used in the preparation of Examples 1-6.

[0022] The tanshinone IIA bilayer polymer soluble acupoint microneedles, which can lower lipids and reduce weight, consist of a needle matrix and a backing matrix. The method and steps are as follows: (1) Needle preparation method: Weigh 10mg of tanshinone IIA and dissolve it fully in 2mL of 60% ethanol-water solution to prepare 5mg / mL tanshinone IIA ethanol-water solution. Weigh 1g of polyvinylpyrrolidone K30 and 1g of chondroitin sulfate and dissolve them in tanshinone IIA ethanol-water solution to prepare needle matrix. Pour the matrix into a soluble acupoint microneedle mold, centrifuge at 4000r / min for 10min and dry at room temperature for 10min to obtain the needle body. The needle body has a needle length of 550µm, a needle array of 137 needles, a thickness of 8mm, an outer diameter of 26mm, and a groove depth of 2.5mm. (2) Preparation method of backing layer: Weigh 75mg tanshinone IIA and dissolve it fully in 15mL 60% ethanol-water solution to prepare 5mg / mL tanshinone IIA ethanol-water solution. Weigh 1.5g polyvinyl alcohol and dissolve it in tanshinone IIA ethanol-water solution to prepare backing matrix. Inject it into a mold containing needles, centrifuge at 4000r / min for 10min, dry at room temperature for 24h, and after demolding, tanshinone IIA soluble acupoint microneedles are obtained.

[0023] Example 2: Preparation of soluble acupoint microneedles made from tanshinone IIA bilayer polymer that can lower lipids and reduce weight. The tanshinone IIA bilayer polymer soluble acupoint microneedles, which can lower lipids and reduce weight, consist of a needle matrix and a backing matrix. The method and steps are as follows: (1) Needle preparation method: Weigh 10mg of tanshinone IIA and dissolve it fully in 2mL of 20% ethanol-water solution to prepare 1mg / mL tanshinone IIA ethanol-water solution. Weigh 1g of polyvinylpyrrolidone K30 and 1g of chondroitin sulfate and dissolve them in tanshinone IIA ethanol-water solution to prepare the needle matrix. Pour the matrix into a soluble acupoint microneedle mold, centrifuge at 3000r / min for 5min and dry at room temperature for 5min to obtain the needle body. The needle body has a needle length of 350µm, a needle array of 137 needles, a thickness of 8mm, an outer diameter of 26mm, and a groove depth of 2.5mm. (2) Preparation method of backing layer: Weigh 75mg tanshinone IIA and dissolve it fully in 15mL 20% ethanol-water solution to prepare 1mg / mL tanshinone IIA ethanol-water solution. Weigh 1.5g polyvinyl alcohol and dissolve it in tanshinone IIA ethanol-water solution to prepare backing matrix. Inject it into a mold containing needles, centrifuge at 3000r / min for 5min, dry at room temperature for 20h, and after demolding, tanshinone IIA soluble acupoint microneedles are obtained.

[0024] Example 3: Preparation of soluble acupoint microneedles made from tanshinone IIA bilayer polymer that can lower lipids and reduce weight. The tanshinone IIA bilayer polymer soluble acupoint microneedles, which can lower lipids and reduce weight, consist of a needle matrix and a backing matrix. The method and steps are as follows: (1) Needle preparation method: Weigh 10mg of tanshinone IIA and dissolve it fully in 2mL of 40% ethanol-water solution to prepare 3mg / mL tanshinone IIA ethanol-water solution. Weigh 1g of polyvinylpyrrolidone K30 and 1g of chondroitin sulfate and dissolve them in tanshinone IIA ethanol-water solution to prepare needle matrix. Pour the matrix into a soluble acupoint microneedle mold, centrifuge at 3500r / min for 8min and dry at room temperature for 8min to obtain the needle body. The needle body has a needle length of 400µm, a needle array of 137 needles, a thickness of 8mm, an outer diameter of 26mm, and a groove depth of 2.5mm. (2) Preparation method of backing layer: Weigh 75mg tanshinone IIA and dissolve it fully in 15mL 40% ethanol-water solution to prepare 3mg / mL tanshinone IIA ethanol-water solution. Weigh 1.5g polyvinyl alcohol and dissolve it in tanshinone IIA ethanol-water solution to prepare backing matrix. Inject it into a mold containing needles, centrifuge at 3500r / min for 8min, dry at room temperature for 24h, and after demolding, tanshinone IIA soluble acupoint microneedles are obtained.

[0025] Example 4: Preparation of soluble acupoint microneedles made from tanshinone IIA bilayer polymer that can lower lipids and reduce weight. The tanshinone IIA bilayer polymer soluble acupoint microneedles, which can lower lipids and reduce weight, consist of a needle matrix and a backing matrix. The method and steps are as follows: (1) Needle preparation method: Weigh 10mg of tanshinone IIA and dissolve it fully in 2mL of 50% ethanol-water solution to prepare 5mg / mL tanshinone IIA ethanol-water solution. Weigh 1g of polyvinylpyrrolidone K30 and 1g of chondroitin sulfate and dissolve them in tanshinone IIA ethanol-water solution to prepare needle matrix. Pour the matrix into a soluble acupoint microneedle mold, centrifuge at 4000r / min for 10min and dry at room temperature for 10min to obtain the needle body. The needle body has a needle length of 450µm, a needle array of 137 needles, a thickness of 8mm, an outer diameter of 26mm, and a groove depth of 2.5mm. (2) Preparation method of backing layer: Weigh 75mg tanshinone IIA and dissolve it fully in 15mL 50% ethanol-water solution to prepare 5mg / mL tanshinone IIA ethanol-water solution. Weigh 1.5g polyvinyl alcohol and dissolve it in tanshinone IIA ethanol-water solution to prepare backing matrix. Inject it into a mold containing needles, centrifuge at 4000r / min for 10min, dry at room temperature for 26h, and after demolding, tanshinone IIA soluble acupoint microneedles are obtained.

[0026] Example 5: Preparation of soluble acupoint microneedles made from tanshinone IIA bilayer polymer that can lower lipids and reduce weight. The tanshinone IIA bilayer polymer soluble acupoint microneedles, which can lower lipids and reduce weight, consist of a needle matrix and a backing matrix. The method and steps are as follows: (1) Needle preparation method: Weigh 10mg of tanshinone IIA and dissolve it fully in 2mL of 60% ethanol-water solution to prepare 8mg / mL tanshinone IIA ethanol-water solution. Weigh 1g of polyvinylpyrrolidone K30 and 1g of chondroitin sulfate and dissolve them in tanshinone IIA ethanol-water solution to prepare needle matrix. Pour the matrix into a soluble acupoint microneedle mold, centrifuge at 4500r / min for 12min and dry at room temperature for 12min to obtain the needle body. The needle body has a needle length of 550µm, a needle array of 137 needles, a thickness of 8mm, an outer diameter of 26mm, and a groove depth of 2.5mm. (2) Preparation method of backing layer: Weigh 75mg tanshinone IIA and dissolve it fully in 15mL 60% ethanol-water solution to prepare 8mg / mL tanshinone IIA ethanol-water solution. Weigh 1.5g polyvinyl alcohol and dissolve it in tanshinone IIA ethanol-water solution to prepare backing matrix. Inject it into a mold containing needles, centrifuge at 4500r / min for 12min, dry at room temperature for 26h, and after demolding, tanshinone IIA soluble acupoint microneedles are obtained.

[0027] Example 6: Preparation of soluble acupoint microneedles made from tanshinone IIA bilayer polymer that can lower lipids and reduce weight. The tanshinone IIA bilayer polymer soluble acupoint microneedles, which can lower lipids and reduce weight, consist of a needle matrix and a backing matrix. The method and steps are as follows: (1) Preparation method of the needle body: Weigh 10 mg of tanshinone IIA and fully dissolve it in 2 mL of 70% ethanol-aqueous solution to prepare a 10 mg / mL ethanol-aqueous solution of tanshinone IIA. Weigh 1 g of polyvinylpyrrolidone K30 and 1 g of chondroitin sulfate and dissolve them in the ethanol-aqueous solution of tanshinone IIA to prepare a needle body matrix, and pour it into a soluble acupoint micro-needle mold. Centrifuge at 5000 r / min for 15 min and then dry at room temperature for 15 min to obtain the needle body. The needle length of the needle body is 650 µm, the needle body array has 137 needles, the thickness is 8 mm, the outer diameter is 26 mm, and the groove depth is 2.5 mm. Set aside for use. (2) Preparation method of the backing layer: Weigh 75 mg of tanshinone IIA and fully dissolve it in 15 mL of 70% ethanol-aqueous solution to prepare a 10 mg / mL ethanol-aqueous solution of tanshinone IIA. Weigh 1.5 g of polyvinyl alcohol and dissolve it in the ethanol-aqueous solution of tanshinone IIA to prepare a backing matrix, and inject it into the mold containing the needle body. Centrifuge at 5000 r / min for 15 min and dry at room temperature for 30 h. After demolding, the soluble acupoint micro-needle of tanshinone IIA is obtained.

[0028] To obtain the solution of the present invention and verify the technical effects of the present invention, the inventors conducted a large number of experimental studies, and some experimental records are as follows: 1. Preparation and evaluation of bilayer polymer soluble acupoint micro-needles loaded with tanshinone IIA 1.1 Experimental animals and materials 1.1.1 Experimental animals SPF-grade C57BL / 6J mice, male, 8 weeks old, body weight (22 ± 2) g, provided by Beijing Speywood Biotechnology Co., Ltd. (license number: SCXK (Beijing) 2019-0010). Six mice were housed in one cage in the Experimental Animal Research Institute of Guizhou University of Traditional Chinese Medicine. They had free access to water and were fed with ordinary feed. The feeding temperature was 22 ± 2 °C, and the indoor lighting was artificially controlled, with 12 h of light (08:00-20:00) and 12 h of darkness (20:00 - 08:00 the next day) alternating in a cycle.

[0029] 1.1.2 Experimental instruments and equipment Table 1 Instruments and equipment 1.1.3 Experimental reagents and drugs Table 2 Reagents and drugs 1.1.4 Technical route Adopt the two-step centrifugation method to prepare soluble acupoint micro-needles of tanshinone IIA, screen out the optimal needle length, determine the drug loading amount, investigate the solubility and safety, and verify acupoint drug administration (see Figure 1 )

[0030] 1.2 Experimental Methods 1.2.1 Preparation of TSA-DMNs The method for preparing tanshinone IIA soluble acupoint microneedles according to claim 1 is characterized in that: the preparation method includes a needle body preparation method and a backing layer preparation method, specifically including the following steps: Needle preparation method: Weigh 10 mg of tanshinone IIA and dissolve it thoroughly in 2 mL of 60% ethanol-water solution to prepare a 5 mg / mL tanshinone IIA ethanol-water solution (see Table 3). Weigh 1 g of polyvinylpyrrolidone K30 and 1 g of chondroitin sulfate and dissolve them in the tanshinone IIA ethanol-water solution to prepare the needle matrix. Pour the matrix into a soluble acupoint microneedle mold, centrifuge at 4000 r / min for 10 min, and then dry at room temperature for 10 min to obtain the needle body for later use.

[0031] Backing layer preparation method: Weigh 75 mg of tanshinone IIA and dissolve it completely in 15 mL of 60% ethanol-water solution to prepare a 5 mg / mL tanshinone IIA ethanol-water solution. Weigh 1.5 g of polyvinyl alcohol and dissolve it in the tanshinone IIA ethanol-water solution to prepare the backing matrix, and inject it into a mold containing needles (see...). Figure 2 Centrifuge at 4000 rpm for 10 min, dry at room temperature for 24 h, and after demolding, obtain tanshinone IIA soluble acupoint microneedles. (Note: Blank DMNs were prepared using the same method without a drug matrix.) Table 3. Screening results for solute concentration and solvent concentration 1.2.2 TSA-DMNs Needle Length Screening 1.2.2.1 Evaluation of the mechanical strength of TSA-DMNs with different needle lengths Cover glass slides were placed on the backing layers of TSA-DMNs with lengths of 350µm, 450µm, 550µm, and 650µm to ensure uniform stress. A 500g weight was placed on the cover plate for about 90 seconds, and the degree of deformation of the needle was observed to examine the mechanical strength of TSA-DMNs with different needle lengths and to eliminate needle lengths with poor mechanical strength.

[0032] 1.2.2.2 Evaluation of the puncture performance of TSA-DMNs with different needle lengths The puncture performance of TSA-DMNs of various needle lengths with good mechanical strength was examined by using an aluminum foil puncture test. The TSA-DMNs were vertically inserted into the aluminum foil for 2 minutes and the puncture results were observed.

[0033] 1.2.2.3 Evaluation of in vitro drug release performance of TSA-DMNs with different needle lengths (1) Establish an HPLC method for detecting TSA content. ①Chromatographic conditions: Agilent Zprbax SB-C18 (250mm×4.68mm, 5μm) column, mobile phase methanol-water (80:20), isocratic elution (1mL / min), wavelength 270nm, column temperature 30℃, injection volume 10µL.

[0034] ②Preparation of TSA reference solution: Accurately weigh 5 mg of TSA reference standard, place it in a 25 mL volumetric flask, dissolve it in methanol and dilute to volume to obtain a TSA reference solution with a mass concentration of 200 μg / mL.

[0035] ③Preparation of TSA-DMNs test solution: Take the prepared TSA-DMNs into a 10mL volumetric flask, add a certain amount of methanol, sonicate for 1h, and then make up to volume. After filtration through a 0.45μm microporous membrane, the solution is obtained.

[0036] ④ Preparation of blank-DMNs test solution: Take the prepared blank-DMNs into a 10mL volumetric flask, add a certain amount of methanol, sonicate for 1h, and then make up to volume. After filtering through a 0.45μm microporous membrane, the solution is obtained.

[0037] ⑤ Specificity test: Take 10 μL each of TSA reference solution, TSA-DMNs test solution and blank-DMNs test solution, and analyze them according to the chromatographic conditions in Part ①. Then compare the chromatograms.

[0038] ⑥ Preparation of standard curve: Take TSA reference solution, dilute with methanol to prepare TSA reference solutions of 10, 20, 40, 80, 100, and 160 μg / mL, and inject and analyze according to the chromatographic conditions in Part ①. Plot the standard curve with TSA mass concentration as the abscissa and peak area as the ordinate.

[0039] ⑦ Precision test: Take the TSA reference solution, dilute it with methanol to prepare an 80 μg / mL TSA reference solution, perform 6 parallel determinations according to the chromatographic conditions in Part ①, record the peak area, and calculate the RSD value.

[0040] ⑧ Stability test: The test solution was injected at 0, 2, 4, 8, 12 and 24 h according to the chromatographic conditions in section 3.1, the peak area was recorded and the RSD value at each time point was calculated.

[0041] ⑨ Repeatability test: Take 6 samples of the same batch of TSA-DMNs, prepare the test solution in parallel according to the method in Part ③, inject continuously according to the chromatographic conditions in Part ①, record the peak area, and calculate the RSD value.

[0042] ⑩ Recovery experiment: Take 9 blank-DMNs and place them in 25mL volumetric flasks. Accurately measure 80, 100, and 160 μg / mL TSA reference solutions respectively and dilute to volume. Inject according to the chromatographic conditions in Part ① to determine the TSA content. Recovery rate = (content of TSA reference solution after adding blank microneedles / content of original TSA reference solution) × 100%.

[0043] (2) Preparation of ex vivo skin Healthy C57BL / 6J mice were euthanized by intraperitoneal injection of sodium pentobarbital 100 mg / kg. Hair was removed from the midline of the abdomen with electric clippers, and the remaining hair was treated with depilatory cream. After washing the skin with warm water, skin of appropriate size was cut off, and subcutaneous fat tissue and fascia were removed. The skin was repeatedly washed with physiological saline, drained, wrapped in aluminum foil, and stored in a refrigerator at -20°C.

[0044] (3) In vitro transdermal test An ethanol / polyethylene glycol-400 / physiological saline solution (volume ratio 5:2:3) was injected into a 7 mL vertical transdermal diffusion cell as the receiving solution. TSA-DMNs were placed on prepared isolated mouse skin (stratum corneum facing upwards), a certain pressure was applied, and the skin was fixed with breathable medical tape. The mouse skin was then placed between the supply cell and the receiving chamber (stratum corneum facing the supply cell). The diffusion cell was then placed on a transdermal diffusion testing instrument, and transdermal tests were conducted at a magnetic stirring speed of 300 r / min and a water bath temperature of 37±1℃. Three parallel tests were performed for each needle length microneedle. 1 mL samples were taken at 0.5, 1, 2, 4, 6, 8, 12, 24, 48, 72, and 96 h (with blank receiving solution replenished). The TSA content was determined by high-performance liquid chromatography (HPLC). The cumulative permeation per unit area Q at each time point was calculated using the following formula: In the formula, Q (µg / cm) 2 ) represents the cumulative permeation per unit area, Cn represents the concentration of each index component measured at the nth sampling point, V represents the volume of the receiving pool (7 mL), Ci represents the concentration of each component measured at all sampling points before the nth sampling point, Vi represents the volume of receiving liquid taken out each time, and A represents the effective contact area in the receiving pool (calculated based on the circular area of ​​the DMNs backing layer, A=πr). 2 =0.785cm 2 After calculation, the cumulative permeation curve of the drug per unit area is plotted with Q as the vertical axis and time t as the horizontal axis.

[0045] 1.2.3 Determination of drug loading in TSA-DMNs Six tablets of the same batch of TSA-DMNs were taken, and the drug loading of TSA-DMNs was determined by HPLC in accordance with the methodology in section 2.2.3. The concentration of the test sample = (sample peak area × reference concentration) / reference peak area, and the drug loading of TSA-DMNs = the concentration of the test sample × the volume of the test sample flask.

[0046] 1.2.4 Investigation of the in vivo solubility of TSA-DMNs Five C57BL / 6J mice were anesthetized and fixed. TSA-DMNs were applied to the midline of the abdomen with a certain force (hair was removed 24 hours before the operation). The needle morphology and dissolution were observed by scanning electron microscopy after 1 min, 10 min, 15 min, 20 min and 25 min respectively.

[0047] 1.2.5 Skin irritation test of TSA-DMNs Twelve C57BL / 6J mice were randomly divided into a blank-DMNs group and a TSA-DMNs group. After routine hair removal, the mice were prepared for 24 hours. Self-control was used: the left abdomen of the mice was treated with either blank-DMNs or TSA-DMNs, while the right abdomen, which was not treated with DMNs, served as a blank control. Treatment was administered every other day for two weeks. The presence of erythema and edema on the skin of the C57BL / 6J mice was recorded after each DMN removal and at 0h, 1h, 24h, 48h, and 72h after the last intervention. Irritation was evaluated according to the Skin Irritation Rating Scale (see Table 4).

[0048] Table 4 Scoring criteria for skin irritation reaction test Note: Skin irritation intensity evaluation criteria: 0-2 mild irritation; 3-5 moderate irritation; 6-8 severe irritation.

[0049] 1.2.6 Comparison of in vitro transdermal performance between acupoint areas and non-acupoint areas 1.2.6.1 Preparation of ex vivo skin Three C57BL / 6J mice were selected to prepare ex vivo skin samples from acupoint areas and non-acupoint areas, respectively. For example... Figure 3 The acupoint shown is Tanzhong (CV17), located according to the Chinese Society of Acupuncture and Moxibustion T / CAAM0002-2020 "Names and Locations of Commonly Used Acupoints in Experimental Animals, Part 3: Mice": on the midline of the abdomen, level with the 4th intercostal space. For non-meridian and non-acupoint points, select 1 cm above the lowest point of the left costal arch, and perform the remaining operations as in section 2.2.3.

[0050] 1.2.6.2 In vitro transdermal assay Follow the procedure for in vitro transdermal experiments in section (3) of “2.2.3”.

[0051] 1.3 Experimental Results 1.3.1 Morphological examination of TSA-DMNs The morphological characterization of TSA-DMNs was observed under both an optical microscope and a scanning electron microscope. For example... Figure 4 As shown (taking 550μm TSA-DMNs as an example), the TSA-DMNs backing is flat, and the 137 needle arrays are arranged in an orderly conical shape on the backing layer. The surface is smooth, the distribution and thickness are uniform, and the needles are sharp.

[0052] 1.3.2 TSA-DMNs Needle Length Screening 1.3.2.1 Evaluation of the mechanical strength of TSA-DMNs with different needle lengths like Figure 5 As shown, 650µm TSA-DMNs are prone to needle breakage and bending under continuous pressure of 500g, while 350µm, 450µm, and 550µm TSA-DMNs remain intact and sharp under continuous pressure of 500g.

[0053] 1.3.2.2 Evaluation of the puncture performance of TSA-DMNs with different needle lengths Due to the poor mechanical strength of 650µm TSA-DMNs, we investigated the puncture performance of TSA-DMNs with needle lengths of 350µm, 450µm, and 550µm. Figure 6 As shown, clear pinhole arrays are visible on the aluminum foils of TSA-DMNs with needle lengths of 350µm, 450µm, and 550µm, and the pinhole arrangement is consistent with the TSA-DMNs needle body.

[0054] 1.3.2.3 Comparison of in vitro drug release performance of TSA-DMNs with different needle lengths (1) Investigation of HPLC methodology ① Specificity examination like Figure 7 As shown, the matrix material does not interfere with the content determination, and the sample and the reference standard have the same chromatographic peaks at corresponding positions in the chromatogram, indicating that the method has good specificity.

[0055] ② Preparation of standard curve like Figure 8 As shown, the linear regression equation is y = 57476x + 8027.3, R0 2 = 0.9999, indicating that TSA has a good relationship within the linear range of 10-160 μg / mL.

[0056] ③Precision test As shown in Table 5, the RSD value calculated based on the peak area results of the 6 peaks is 0.167%, which is less than 3%, indicating that the method has good precision.

[0057] Table 5 Precision test results ④ Stability test As shown in Table 6, the peak area RSD of the TSA-DMNs test solution was measured to be 0.121%, which is less than 3%, indicating that the method has good stability.

[0058] Table 6 Stability test results ⑤ Repeatability experiment As shown in Table 7, the peak area RSD of the TSA-DMNs test solution was 0.061%, which is less than 3%, indicating that the method has good repeatability.

[0059] Table 7 Results of Repeatability Experiments ⑥ Recovery rate experiment As shown in Table 8, the measured recoveries were 101.251, 99.333, and 101.225, respectively, with an RSD of 1.095%, which is less than 3%, indicating that the recovery rate of this method meets the requirements.

[0060] Table 8 Results of the spiking recovery experiment (2) In vitro transdermal test Because 650µm TSA-DMNs have poor mechanical strength, while 350µm, 450µm, and 550µm TSA-DMNs have good mechanical strength and puncture performance, we investigated the drug release performance of 350µm, 450µm, and 550µm TSA-DMNs. The results are as follows: Figure 9 As shown: The drug release rates of TSA-DMNs at 350µm, 450µm, and 550µm were relatively high in the 0-24h timeframe. 0.5h The values ​​were 150.67 ± 75.45 μg / cm³. 2 179.65±81.63μg / cm 2 192.99±69.93μg / cm 2 Q 6h The values ​​were 348.93 ± 82.93 μg / cm³. 2 379.63±83.42μg / cm 2 407.79±92.48μg / cm 2 Q 12h The values ​​were 475.53 ± 86.78 μg / cm³. 2 519.75±95.38μg / cm2 537.30±93.81μg / cm 2 .

[0061] The drug release rate of TSA-DMNs at 350µm, 450µm, and 550µm in the 24-48h time frame is relatively slow. 24h They were 801.40±107.38 μg / cm³, respectively. 2 848.74±107.51μg / cm 2 866.76±102.65μg / cm 2 Q 48h They were 1097.84±101.94 μg / cm³, respectively. 2 1167.26±89.63μg / cm 2 1211.70±78.34μg / cm 2 .

[0062] The relative equilibrium time for drug release of 350µm, 450µm, and 550µm TSA-DMNs was around 48 hours, but the cumulative permeation per unit area of ​​550µm TSA-DMNs was consistently higher than that of 350µm and 450µm TSA-DMNs.

[0063] 1.3.3 Determination of drug loading in TSA-DMNs As shown in Table 9, the average drug loading of TSA-DMNs is 1.32 ± 0.02 mg / tablet.

[0064] Table 9. Determination of drug loading in TSA-DMNs 1.3.4 Investigation of the in vivo solubility of TSA-DMNs like Figure 10 As shown, when 550µm TSA-DMNs are inserted into the skin and come into contact with the interstitial fluid, the needle tip begins to dissolve. After 10 minutes of insertion, it begins to dissolve in most parts. After 15 minutes, two-thirds of the needle body is completely dissolved. After 25 minutes, except for the part of the needle body near the backing layer, the needle body is almost completely dissolved. 1.3.5 Skin irritation test of TSA-DMNs No obvious symptoms such as erythema, edema, or skin elevation were observed in C57BL / 6J mice in the blank-DMNs group and the TSA-DMNs group after each removal of DMNs and at 0h, 1h, 24h, 48h, and 72h after the last intervention. The skin irritation response scores were all 0, indicating that the TSA and DMNs matrix materials were non-irritating to the mouse skin.

[0065] 1.3.6 Comparison of in vitro transdermal performance between acupoint areas and non-acupoint areas like Figure 11 As shown: In the 0-24h interval, TSA-DMNs showed relatively high drug release rates at the Tanzhong acupoint and non-meridian, non-acupoint points. 0.5h They were 242.16 ± 68.25 μg / cm³, respectively. 2 202.32±66.81μg / cm 2 Q 6h The values ​​were 449.94 ± 87.55 μg / cm³. 2 390.87±82.68μg / cm 2 Q 12h The values ​​were 578.31 ± 84.44 μg / cm³. 2 526.77±96.19μg / cm 2 .

[0066] The release rate of TSA-DMNs at the Tanzhong acupoint and non-meridian / non-acupoint points was relatively slow during the 24-48h interval. 24h The values ​​were 913.51 ± 107.70 μg / cm³. 2 862.00±99.61μg / cm 2 Q 48 They were 1259.06±64.50 μg / cm³, respectively. 2 1178.20±87.37μg / cm 2 .

[0067] The relative equilibrium time for TSA-DMNs release at both the Tanzhong acupoint and non-meridian / non-acupoint points is around 48 hours, but the cumulative permeation per unit area of ​​the skin region at the Tanzhong acupoint is consistently higher than that at non-meridian / non-acupoint points.

[0068] 1.4 Summary (1) In this study, TSA-DMNs prepared by using 5 mg / L TSA ethanol-water solution as raw material, PVP K30 and CS as needle matrix materials and PVA as backing matrix material showed good morphological characterization.

[0069] (2) Under the above preparation conditions, the mechanical strength of 650µm TSA-DMNs is poor, while the mechanical strength and puncture performance of 350µm, 450µm and 550µm TSA-DMNs are good.

[0070] (3) The relative equilibrium time of drug release of 350µm, 450µm and 550µm TSA-DMNs is about 48h. Among them, the drug release performance of 550µm TSA-DMNs is better than that of 350µm and 450µm TSA-DMNs. (4)The relative drug release equilibrium time of 550 µm TSA-DMNs on the skin of the Tanzhong acupoint and non-acupoint and non-meridian points was about 48 h, and the skin permeability in the Tanzhong acupoint area was better than that in non-acupoint and non-meridian points.

[0071] The results showed that 550 µm TSA-DMNs had good mechanical strength and puncture performance, and had the best drug release performance, reaching drug release equilibrium in about 48 h. This experiment also confirmed the superiority of TSA-DMNs in acupoint drug delivery, providing in vitro experimental evidence for the establishment of the composite drug delivery system DAMNs.

[0072] 2. Improvement of blood lipid and body weight levels in mice by tanshinone IIA double-layer polymer soluble acupoint micro needles 2.1 Experimental animals and materials 2.1. Experiment animals SPF-grade C57BL / 6J mice and C57BL / 6J background ApoE - / - mice, male, 8 weeks old, body weight (22 ± 2) g, provided by Beijing Speyford Biotechnology Co., Ltd. (license number: SCXK (Beijing) 2019-0010), housed in cages of 6 each in the Experimental Animal Research Institute of Guizhou University of Traditional Chinese Medicine, with free access to water and fed with ordinary feed, the feeding temperature was 22 ± 2 °C, the indoor lighting was artificially controlled, and the 12 h light (08:00-20:00) and 12 h darkness (20:00 - 08:00 the next day) were alternated in cycles. The study was reviewed by the Experimental Animal Ethics Committee of Guizhou University of Traditional Chinese Medicine (No: 20230009).

[0073] 2.1.2 Experimental instruments and equipment Table 10 Instruments and equipment 2.1.3 Experimental reagents and drugs Table 11 Reagents and drugs 2.2 Experimental methods 2.2.1 Modeling method and evaluation After 1 week of adaptive feeding with ordinary feed, 6 C57BL / 6J mice were fed with normal diet, and 18 C57BL / 6J background ApoE - / - mice were fed continuously with high-fat feed (formulation: 38.4% carbohydrate, 22.5% fat, 21.3% protein, 5.7% crude fiber, 1.8% calcium, 1.2% phosphorus, prepared by Beijing Botai Hongda Biotechnology Co., Ltd.) for 8 weeks to prepare a hyperlipidemia model.

[0074] 2.2.2 Grouping Six C57BL / 6J mice were set as the blank control group, and 18 ApoE mice that successfully developed the model were used. - / - Mice were randomly divided into a model group, an acupoint microneedling group, and a blank microneedling group using a random number table method, with 6 mice in each group.

[0075] 2.2.3 Intervention Methods 2.2.3.1 Blank Group Feed with regular feed, hold and fix the food for 20 minutes, once a day, for 8 consecutive weeks.

[0076] 2.2.3.2 Model Group Feed with regular feed, hold and fix the food for 20 minutes, once a day, for 8 consecutive weeks.

[0077] 2.2.3.3 Acupoint Microneedle Group Mice were fed a standard diet, held and immobilized for 20 minutes, once daily for 8 weeks. Additionally, each mouse underwent local hair removal and routine disinfection, followed by intervention at the Tanzhong acupoint, with appropriate pressure applied. One tablet was administered each time, with the location following the guidelines of the China Association of Acupuncture and Moxibustion T / CAAM 0002-2020 "Common Acupoint Names and Locations in Laboratory Animals Part 3: Mice" (see [link to relevant documentation]). Figure 12 Once every other day for 8 consecutive weeks.

[0078] 2.2.3.4 Blank microneedle group Feed the animals with regular feed, hold and fix them for 20 minutes, once daily for 8 consecutive weeks. During the fixation process, perform blank-DMNs intervention at the Tanzhong acupoint, following the procedure and treatment course of "item 2.2.3.3 acupoint microneedling group".

[0079] 2.2.4 Material Sampling Method After the final intervention, the mice were fasted for 12 hours but allowed free access to water, and blood was collected. Using enucleation, approximately 1-1.5 mL of blood was collected from the orbital venous plexus of the mice and placed in 2 mL EP tubes. After standing at room temperature for 1 hour, the tubes were centrifuged at 3000 rpm for 10 minutes at 4°C. The serum was then stored at -80°C for later use in lipid analysis.

[0080] 2.2.5 Observation Indicators and Detection Methods The serum levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) in each group of mice were measured. Serum samples were pretreated, centrifuged at 3000 rpm for 10 min, and the supernatant was collected for analysis. Results were exported from a fully automated biochemical analyzer.

[0081] 2.2.6 Data Statistics Data were analyzed using SPSS 26.0 software. Normally distributed continuous data were expressed as mean ± standard deviation (SD). Data presented as follows: If variances are homogeneous, a one-way ANOVA test is used to examine differences between groups; multiple comparisons are performed using the LSD method. If variances are unequal, a corrected Welch ANOVA test is used to examine differences between groups; multiple comparisons are performed using the Games-Howell method. Non-normally distributed continuous data are presented as median (interquartile range) M (IQR), and the Kruskal-Wallis H test is used. P < 0.05 is considered statistically significant.

[0082] 2.3 Experimental Results 2.3.1 General Observation The mice in the blank control group were in good spirits, active normally, with shiny and smooth fur, and normal diet and water intake. The mice in the model group and blank microneedle group were in poor spirits, with reduced activity, greasy fur, and some mice had localized hair loss. Their diet was slightly poor, but their water intake was normal. The mice in the acupoint microneedle group were in good spirits, with slightly greasy fur. Some mice had localized hair loss, but their diet and water intake were normal.

[0083] 2.3.2 Weight Comparison As shown in Table 12: Before treatment (after modeling): Compared with the blank group, the body weight of mice in all other groups increased (P<0.001), and there was no statistically significant difference in body weight among the model group, acupoint microneedling group and blank microneedling group (P>0.05).

[0084] After treatment: Compared with the model group, the weight of mice in the acupoint microneedling group was reduced (P<0.01), while there was no statistically significant difference in weight of mice in the blank microneedling group (P>0.05).

[0085] Compared with the acupoint microneedling group, the mice in the blank microneedling group had increased body weight (P<0.05).

[0086] Table 12 Comparison of mouse body weight among groups Note: Compared with the blank group, *** P < 0.001; compared with the model group, △△ P<0.01; compared with the acupoint microneedling group, # P < 0.05.

[0087] 2.3.3 Comparison of serum TC, TG, HDL-C, and LDL-C levels As shown in Tables 13 and 14: Compared with the control group, the model group mice showed increased levels of TC, TG, and LDL-C (P<0.001), and decreased levels of HDL-C (P<0.001).

[0088] Compared with the model group, the levels of TC, TG, and LDL-C in the acupoint microneedle group decreased (P<0.001), while the level of HDL-C increased (P<0.001). In contrast, there was no statistically significant difference in the levels of TC, TG, LDL-C, and HDL-C in the blank microneedle group (P>0.05). Compared with the acupoint microneedling group, mice in the blank microneedling group showed increased levels of TC, TG, and LDL-C (P<0.001), and decreased levels of HDL-C (P<0.001).

[0089] Table 13 Comparison of TC and TG levels in mice of different groups Note: Compared with the blank group, *** P < 0.001; compared with the model group, △△△ P<0.001; compared with the acupoint microneedling group, ### P < 0.001, ## P < 0.01.

[0090] Table 14 Comparison of HDL-C and LDL-C levels in mice of different groups Note: Compared with the blank group, *** P < 0.001; compared with the model group, △△△ P<0.001, △ P<0.05; compared with the acupoint microneedling group, ### P < 0.001, # P < 0.05.

[0091] 2.4 Summary (1) ApoE- / - mice, a hyperlipidemia model established by a high-fat diet, exhibited poor mental state and dietary condition. Tanshinone IIA bilayer polymer-soluble acupuncture points can improve the hyperlipidemia model ApoE- / -. - / - The study observed changes in the mice's mental state and diet, but it did not significantly improve hair loss caused by a high-fat diet.

[0092] (2) ApoE, a hyperlipidemia model established by a high-fat diet - / - Mice maintained a balanced body weight. Tanshinone IIA bilayer polymer-soluble acupoint microneedles were able to reduce ApoE in a hyperlipidemic model. - / - Mouse body weight, while blank bilayer polymer soluble acupoint micro-targets in the hyperlipidemia model ApoE - / - There was no significant effect on mouse body weight.

[0093] (3) ApoE, a hyperlipidemia model established by a high-fat diet - / - Mice exhibited abnormal lipid metabolism. Tanshinone IIA bilayer polymer-soluble acupoint microneedles were able to improve ApoE in a hyperlipidemic model. - / - The levels of four lipid parameters in mice, and the use of blank bilayer polymer-soluble acupoint micro-targets in a hyperlipidemia model ApoE - / - It did not significantly improve abnormal lipid metabolism in mice.

[0094] The results of this study indicate that the tanshinone IIA bilayer polymer soluble acupoint microneedles have a definite effect on regulating body weight and reducing blood lipids, while the blank bilayer polymer soluble acupoint microneedles do not have such effects.

Claims

1. A tanshinone ⅡA double-layer polymer soluble acupoint microneedle for reducing fat and weight loss, which is composed of a needle body matrix and a backing layer matrix, characterized in that: The needle body matrix is prepared from 1-10 parts of tanshinone IIA, 1000-5000 parts of polyvinyl pyrrolidone K30 and 1000-5000 parts of chondroitin sulfate, and the backing layer matrix is prepared from 20-75 parts of tanshinone IIA and 5000-15000 parts of polyvinyl alcohol. ​ 2. The tanshinone IIA double-layer polymer soluble acupoint microneedle according to claim 1, characterized in that: The needle body matrix is prepared from 5-10 parts of tanshinone IIA, 1000-3000 parts of polyvinyl pyrrolidone K30 and 1000-3000 parts of chondroitin sulfate, and the backing layer matrix is prepared from 50-75 parts of tanshinone IIA and 8000-15000 parts of polyvinyl alcohol.

3. The tanshinone IIA double-layer polymer soluble acupoint microneedle according to claim 2, characterized in that: The needle body matrix is prepared from 10 parts of tanshinone IIA, 1000 parts of polyvinyl pyrrolidone K30 and 1000 parts of chondroitin sulfate, and the backing layer matrix is prepared from 75 parts of tanshinone IIA and 15000 parts of polyvinyl alcohol.

4. The tanshinone IIA double-layer polymer soluble acupoint microneedle according to any one of claims 1-3, characterized in that: The tanshinone IIA is first dissolved in a 20-70% ethanol-water solution, and then the needle body matrix and the backing layer matrix are prepared, and the concentration of tanshinone IIA in the tanshinone IIA ethanol-water solution is 1-7 mg / mL.

5. The tanshinone IIA double-layer polymer soluble acupoint microneedle according to claim 4, characterized in that: The tanshinone IIA is first dissolved in a 60% ethanol-water solution, and then the needle body matrix and the backing layer matrix are prepared, and the concentration of tanshinone IIA in the tanshinone IIA ethanol-water solution is 5 mg / mL.

6. The preparation method of tanshinone ⅡA double-layer polymer soluble acupoint microneedle according to any one of claims 1-6, characterized in that: The preparation method comprises a needle body preparation method and a backing layer preparation method, and specifically comprises the following steps: (1) Needle body preparation method: 10 mg of tanshinone IIA is weighed and dissolved in 2 mL of a 20-70% ethanol-water solution to prepare a tanshinone IIA ethanol-water solution with a concentration of 1-10 mg / mL, 1 g of polyvinyl pyrrolidone K30 and 1 g of chondroitin sulfate are weighed and dissolved in the tanshinone IIA ethanol-water solution to prepare a needle body matrix, and then the needle body matrix is poured into a soluble acupoint microneedle mold, centrifuged at 3000-5000 r / min for 5-15 min, and dried at room temperature for 5-15 min to obtain the needle body, wherein the needle length of the needle body is 350 µm-650 µm, the needle body array has 137 needles, the thickness is 8 mm, the outer diameter is 26 mm, the groove depth is 2.5 mm, and the needle body is ready for use; (2) Backing layer preparation method: 75 mg of tanshinone IIA is weighed and dissolved in 15 mL of a 20-70% ethanol-water solution to prepare a tanshinone IIA ethanol-water solution with a concentration of 1-10 mg / mL, 1.5 g of polyvinyl alcohol is weighed and dissolved in the tanshinone IIA ethanol-water solution to prepare a backing matrix, and then the backing matrix is injected into the mold containing the needle body of step (1), centrifuged at 3000-5000 r / min for 5-15 min, and dried at room temperature for 20-30 h, and then demolded to obtain the tanshinone IIA soluble acupoint microneedle.

7. The preparation method of tanshinone IIA double-layer polymer soluble acupoint microneedle according to claim 8, characterized in that: The preparation method comprises a needle body preparation method and a backing layer preparation method, and specifically comprises the following steps: (1) The preparation method of the needle body: 10 mg of tanshinone IIA is fully dissolved in 2 mL of a 60% ethanol-water solution to prepare a tanshinone IIA ethanol-water solution of 5 mg / mL. 1 g of polyvinyl pyrrolidone K30 and 1 g of chondroitin sulfate are dissolved in the tanshinone IIA ethanol-water solution to prepare a needle body matrix, and then poured into a soluble acupoint microneedle mold. After centrifugation at 4000 r / min for 10 min, the needle body is dried at room temperature for 10 min. The needle length of the needle body is 550 µm, the needle body array is 137, the thickness is 8 mm, the outer diameter is 26 mm, the groove depth is 2.5 mm, and the needle body is ready for use; (2) The preparation method of the backing layer: 75 mg of tanshinone IIA is fully dissolved in 15 mL of a 60% ethanol-water solution to prepare a tanshinone IIA ethanol-water solution of 5 mg / mL. 1.5 g of polyvinyl alcohol is dissolved in the tanshinone IIA ethanol-water solution to prepare a backing matrix, and then injected into the mold containing the needle body. After centrifugation at 4000 r / min for 10 min, the tanshinone IIA soluble acupoint microneedle is dried at room temperature for 24 h, and then demolded.

8. The application of the tanshinone IIA double-layer polymer soluble acupoint microneedle in improving the preparation of hyperlipidemia.

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