Guanxin storax pill volatile oil nano-liposome / PVA swelling type microneedle patch

Abstract

The invention belongs to the technical field of acupuncture point patch preparation, and relates to a coronary heart storax pill volatile oil nano-liposome / PVA swelling type microneedle patch. Crushing storax, frankincense, sandalwood and elecampane, adding water for soaking, slightly boiling, condensing and refluxing; and drying to obtain the GXSHW-EO. Soybean lecithin, cholesterol, sodium cholate and GXSHW-EO are added into methyl alcohol and trichloromethane, a phosphoric acid buffer solution is added after rotary evaporation, ultrasonic treatment and filtering are conducted after rotary evaporation, and the microneedle patch is obtained after a PVA solution, a PVP solution and a citric acid solution are evenly mixed, subjected to centrifugal vacuum pouring, vacuumized, dried and demoulded. By integrating the targeting delivery advantage of the NLCs, the intelligent swelling characteristic of the PVA microneedle patch and the meridian regulation effect of acupoint stimulation, the triple synergistic effect of physical infiltration promotion, chemical drug release and biological response is achieved. The drug loading capacity and the stability of active ingredients of the traditional Chinese medicine are enhanced.

Description

Technical Field

[0001] The present invention belongs to the technical field of acupoint patch preparation, and relates to a nanoliposome of volatile oil of Guanxin Suhe Pills / PVA swelling microneedle patch. Background Art

[0002] Acupoint application therapy exerts local or overall regulatory effects through the meridian system. According to factors such as the patient's constitution, skin sensitivity, and the magnitude of drug stimulation, the appropriate application time is selected, and the bidirectional regulatory effect of acupoints, the continuous stimulation of acupoints by drugs, and the combined effect of both are utilized. It achieves the purpose of preventing and treating diseases by regulating aspects such as the body's skin immune system, skin microbiome, and circulatory system. The outermost layer of the human meridian system is the skin area, which is an important link for the action force of acupoint application drugs and the reaction of zang-fu functions, directly determining the penetration rate, direction, and effectiveness of the drug. The main barrier for drug percutaneous penetration and absorption in acupoint application therapy is the stratum corneum of the skin.

[0003] In traditional powder acupoint application therapy, the transdermal absorption rate of drugs is relatively low, the adhesion stability is poor, and there is also a problem of difficult dose control, which affects the stability of the curative effect. Summary of the Invention

[0004] The present invention proposes a novel nanoliposome of volatile oil of Guanxin Suhe Pills / PVA swelling microneedle patch for the problems existing in traditional powder acupoint application therapy.

[0005] In order to achieve the above purpose, the present invention is realized by adopting the following technical solutions: A preparation method of a nanoliposome of volatile oil of Guanxin Suhe Pills / PVA swelling microneedle patch, the steps are as follows: (1) Weigh the following raw materials by weight parts: 45 - 55 parts of styrax, 100 - 110 parts of frankincense, 205 - 215 parts of sandalwood, 205 - 215 parts of costus root. After being crushed respectively and passing through a 100 - mesh sieve and mixing evenly, a mixture A is obtained; add deionized water, seal and soak, then place it in a device with heating and condensing reflux, and keep it in a slightly boiling and condensing reflux state for at least 12 h; after standing, absorb the oil layer and add a desiccant to dry, to obtain the volatile oil GXSHW - EO of Guanxin Suhe Pills.

[0006] (2) Add soybean lecithin, cholesterol, sodium cholate, GXSHW - EO to a mixed solvent of methanol and chloroform, mix evenly and then perform rotary evaporation; after the rotary evaporation is completed, add a phosphate buffer solution, perform rotary evaporation again and then ultrasonic treatment and filtration to obtain the nanoliposome solution GXSHW - EO - NLCs of the volatile oil of Guanxin Suhe Pills.

[0007] (3) Mix the PVA solution, PVP solution, citric acid solution, and GXSHW-EO-NLCs solution, and centrifuge to obtain the microneedle solution; pour the microneedle solution into a PDMS mold by vacuum casting, evacuate, dry, and demold to obtain the swelling microneedle patch.

[0008] Preferably, in step (1), the mass of deionized water added is 5.8 - 6.2 times the mass of mixture A, the sealing immersion time is 11 - 13 h, and the desiccant is anhydrous sodium sulfate.

[0009] Preferably, in step (2), the mass ratio of soybean lecithin to GXSHW-EO is (3.9 - 4.1):1, the mass ratio of soybean lecithin to cholesterol is (5.9 - 6.1):1, and the mass of sodium cholate is 2.0 - 3.0 mg; the volume ratio of methanol to chloroform in the mixed solvent is 1:1; the rotary evaporation speed is 150 - 170 rpm, the rotary evaporation temperature is 45 - 55 °C, and the pH of the phosphate buffer solution is 6.7 - 6.9.

[0010] Preferably, in step (2), the filtration is carried out using a 0.45 μm microporous membrane.

[0011] Preferably, in step (2), the mass fraction of the PVA solution is 3.5 - 4%, the mass fraction of the PVP solution is 5 - 5.5%, the mass fraction of the citric acid solution is 0.3 - 0.5%, and the mass fraction of the GXSHW-EO-NLCs solution is 15 - 20%; the volume ratio of the PVA solution:PVP solution:citric acid solution:GXSHW-EO-NLCs solution is (29 - 31):(33 - 36):(4 - 6):(29 - 31).

[0012] Preferably, in step (3), the bottom radius of the swelling microneedle patch is 120 - 123 μm, the height of the needle body is 500 - 502 μm; the angle of the microneedle tip is 29 - 32 μm, the tip spacing is 560 - 580 µm, and the number of needle bodies in each microneedle patch is 140 - 150.

[0013] Principle: The traditional preparation method of acupoint application is to grind traditional Chinese medicine into fine powder, sieve it through 80 - 100 meshes, and mix it with adjuvants (such as ginger juice, honey) in proportion to make a thick paste, with the degree that it does not drip when picked up. During treatment, take the ointment and knead it into the size of soybeans to broad beans, place it in the center of the gauze, with a thickness of about 2 - 3 millimeters, directly apply it to the acupoint, and fix it with adhesive tape. The application time depends on the specific situation. Because some drugs (such as white mustard seed, asarum, and mylabris) are likely to cause skin redness, blisters, and even allergic reactions. In the present invention, first, the traditional Chinese medicine is crushed and sieved through 100 meshes, the volatile oil contained in the traditional Chinese medicine is extracted by steam distillation method, then the extracted traditional Chinese medicine volatile oil is loaded into nanoliposomes, and finally, the nanoliposomes loaded with traditional Chinese medicine volatile oil are mixed with materials such as PVA to prepare the volatile oil nanoliposome / PVA swelling type microneedle patch of Guanxin Suhe Pills. During treatment, directly apply the microneedle patch to the acupoint, fix it with adhesive tape, and apply it for 12 hours. During the application period, the irritation to the skin is extremely small.

[0014] Compared with the prior art, the advantages and positive effects of the present invention are as follows: The present invention constructs the GXSHW-EO-NLCs / PVA swelling type microneedle patch, and realizes the triple synergistic effect of "physical penetration enhancement - chemical drug release - biological response" by integrating the targeted delivery advantage of NLCs, the intelligent swelling property of PVA microneedle patch, and the meridian regulation effect of acupoint stimulation. The prepared microneedle patch can achieve accurate quantitative drug delivery and sustained and controlled release of drugs, improve the transdermal permeability of drugs, enhance the drug loading capacity and the stability of active ingredients of traditional Chinese medicine, and at the same time can significantly improve the stability and biocompatibility of the patch, and improve the disadvantages of the traditional acupoint application dosage form such as unclear transdermal absorption rate and easy pollution of clothes, so as to achieve the curative effect of "combination of acupuncture and medicine" in the treatment of MI. Brief Description of the Drawings

[0015] Figure 1 It is the puncture experimental diagram of the swelling type microneedle patch prepared in Example 1. Detailed Embodiments

[0016] In order to more clearly understand the above objects, features and advantages of the present invention, the following further describes the present invention with specific embodiments. It should be noted that, without conflict, the embodiments of the present application and the features in the embodiments can be combined with each other.

[0017] Many specific details are set forth in the following description to fully understand the present invention. However, the present invention can also be implemented in other ways different from those described herein. Therefore, the present invention is not limited by the specific embodiments disclosed in the following specification.

[0018] Example 1 Weigh 50 g of storax, 105 g of frankincense, 210 g of sandalwood, and 210 g of costus root. Use a pulverizer to crush them into 100-mesh fine powder multiple times. After sieving, mix them evenly and place them in a round-bottom flask. Add 6 times the mass of deionized water, stir evenly with a glass rod, seal and soak for 12 h. After the soaking is completed, put the flask into an electric heating mantle, stir evenly with a glass rod again, then connect a glass condenser to the upper mouth of the flask, and connect flowing tap water as the condensation medium. Turn on the power of the condensation reflux device and the electric heating mantle. When the liquid in the flask is slightly boiling, adjust the temperature and maintain it at the boiling state for 12 h. Turn off the power and wait for the volatile oil determination device to cool naturally to room temperature. Then disassemble the reflux device, read the scale of EO, suck out EO, add 5 g of anhydrous sodium sulfate for drying to obtain GXSHW-EO.

[0019] Precisely weigh 12 g of soybean phospholipid, 2 g of cholesterol, 2.5 mg of sodium cholate, and 3 g of GXSHW-EO, and dissolve them in a mixed solvent containing 10 mL of methanol and 10 mL of chloroform. Seal it with a sealing film and stir magnetically until completely dissolved, then place it in a 500-mL round-bottom flask. Install the round-bottom flask on a rotary evaporator, set the rotation speed to 160 rpm, and perform vacuum distillation in a 50°C constant temperature water bath for 2 h to volatilize the organic solvent. At the same time, a uniform and transparent lipid film appears on the inner wall of the round-bottom flask. After turning off the rotary evaporator, add 20 mL of phosphate buffer solution (pH 6.8) to the round-bottom flask, and continue to rotate the elution device to wash the lipid film material on the wall to make the film fall off to obtain primary liposomes. After 80 min, remove the round-bottom flask, seal and ultrasonicate for 40 min to obtain a liposome suspension. Let it stand at room temperature for 10 min and filter it with a 0.45-μm microporous filter membrane to obtain a GXSHW-EO-NLCs solution with a white opalescence.

[0020] Polyvinyl alcohol (PVA) with a molecular weight of 89 - 98 kDa and polyvinylpyrrolidone (PVP) with a molecular weight of 1300 kDa were selected as the matrix materials for the microneedles. 0.4 g of PVA was added to a sterile and enzyme - free centrifuge tube, 10 mL of deionized water was added, stirred with a glass rod and then the lid was tightened. It was placed in an 80 °C electro - thermal blast drying oven and stirred with a glass rod every 30 min. After 2 h, the PVA was completely dissolved to form a homogeneous PVA solution. After natural cooling, it was reserved for use. Preparation method of the PVP solution: 0.56 g of PVP was added to a sterilized beaker, 10 mL of deionized water was added, stirred with a glass rod, then a rotor was put in, the beaker was sealed with a sealing film, and stirred thoroughly in a magnetic stirrer at room temperature for 2 h to obtain a homogeneous PVP solution. Preparation method of the citric acid solution: 0.04 g of citric acid was added to a sterilized beaker, 10 mL of deionized water was added, a rotor was put in, the beaker was sealed with a sealing film, and stirred thoroughly in a magnetic stirrer at room temperature for 2 h to obtain a homogeneous citric acid solution.

[0021] 3 mL of the PVA solution, 3.5 mL of the PVP solution, 0.5 mL of the citric acid solution and 3 mL of the GXSHW - EO - NLCs solution were mixed evenly, and centrifuged at 4000 rpm for 15 min to remove air bubbles. The GXSHW - EO - NLCs / PVA swelling microneedle patch was prepared by the vacuum casting method. The obtained mixed liquid was cast into a PDMS mold (which can be commercially customized according to the specifications of the microneedle patch and the requirements of vacuum pumping), vacuum - pumped at 0.90 ± 0.05 MPa for 15 min, dried at 30 °C for 2 h, and the GXSHW - EO - NLCs / PVA swelling microneedle patch was obtained after demolding.

[0022] After testing, the bottom radius of the swelling microneedle patch obtained in this example was 121.39 µm, the needle body height was 501.22 µm, the microneedle tip angle was 29.93 µm, the tip spacing was 570 µm, and the number of needle bodies of the microneedle patch was 145. After further testing, the cinnamic acid content of each swelling microneedle patch obtained in this example was 10.98 µg, and the costunolide content was 0.21 mg.

[0023] 1. Skin penetration performance test SPF-grade SD rats provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. (Animal License No.: SCXK (Beijing) 2016-0006), 6 weeks old, with a body weight of 200 ± 20 g. After the rats were completely anesthetized, their four limbs were fixed with medical tape. The hair on the back was removed with a hair clipper and then depilatory cream was applied. After standing for 3 min, it was wiped with a gauze soaked in pure water and then dried with a cotton ball. The swelling-type microneedle patch prepared in Example 1 was pressed into the back skin of the rats with a microneedle assistor (20 N / cm 2 ), and after 1 min, the microneedle patch was removed. The rats were sacrificed, and the skin of the rats at the microneedle patch application site was quickly removed. It was unfolded and flattened on a dissection board, and the skin edge was fixed with medical tape. Immediately, a 0.1% trypan blue solution was evenly dropped onto the skin area pricked by the microneedle patch, and the staining was maintained for 20 min. After the staining was completed, the excessive trypan blue solution was blotted with a dry cotton ball, and then wiped with a cotton ball dipped in physiological saline. The results are as Figure 1 shown. As can be seen from Figure 1 , clear pores caused by the tiny needles indenting into the skin appeared on the rat skin, presenting regularly arranged blue dots, proving that the microneedle patch prepared in Example 1 could effectively penetrate the skin.

[0024] 2. Skin recovery test Randomly select 5 healthy SPF-grade SD rats. After anesthesia, the hair on their backs was removed with a hair clipper and depilatory cream was evenly applied. After standing for 2 min, it was wiped clean. The swelling-type microneedle patch of Example 1 was pressed into the back skin of the rats (20 N / cm 2 ), and fixed with medical tape for 3 min. Then the tape and the microneedle patch were removed, and the recovery condition of the rat skin was observed. After detection, tiny cavity structures were basically generated at the microneedle patch application site during normal use, which were the same as the microneedle array, and slightly red appeared locally. During the 30 - 120 min of microneedle patch application, it was obvious that the depth and width of the cavity structure on the skin surface gradually decreased, and the skin color at the application site gradually tended to the normal skin color around. At 120 min, the skin healing was basically completed, it was difficult to observe the existence of the cavity, and the skin color completely returned to the normal state. Thus, it can be known that the microneedle patch prepared in Example 1 has good biocompatibility, causes less damage to the skin, and does not affect skin recovery.

[0025] 3. Investigation of the solubility of the needle body Randomly select 5 healthy SPF-grade SD rats. After anesthetizing the rats, use a hair clipper to remove the hair on their backs, apply depilatory cream and let it stand for 3 min, then wipe it clean with a wet gauze and a dry cotton ball. Apply 4 swelling micro-needle patches prepared in Example 1 to the back skin of each rat respectively, and fix them with medical adhesive tape on the upper part. Remove one patch at 0.5 h, 1 h, 1.5 h, and 2 h respectively, and observe the dissolution of the needles in the micro-needle patch with a microscope and record it. After detection, after removing the patches at 0.5 h, 1 h, 1.5 h, and 2 h, the needles of the micro-needle patches are slightly bent, and with the passage of time, swelling occurs gradually due to absorbing the surrounding tissue fluid. The micro-needle bodies are still not dissolved at 2 h, indicating that the micro-needles prepared in this study have good swelling ability and certain sustained transdermal drug release ability.

[0026] 4. In vitro transdermal test After the rats are completely anesthetized, use a hair clipper to shave the abdominal hair of the rats, then apply depilatory cream to the depilated abdominal area, let it stand for 5 min, wipe the local skin clean with a wet gauze and a dry cotton ball. After sacrificing the rats, carefully remove the skin, carefully strip and remove the adipose tissue and fascia of the skin. After washing it several times with physiological saline, dry the skin surface with a dry filter paper, flatten the skin and wrap it with tin foil, and put it in a -20 °C refrigerator for standby. Take out the rat skin wrapped with tin foil from the -20 °C refrigerator, put the skin into physiological saline, soak it at room temperature for 30 min, then rinse it carefully with physiological saline and set it aside. It is necessary to carefully check the integrity of the rat skin, and do not use damaged skin. Dry the water on the skin surface with a filter paper, cut an appropriate area to prepare an excised abdominal rat skin with the same area as the diffusion cell. Inject 30% ethanol physiological saline into the receiving chamber of the Franz diffusion cell as the receiving solution, and there should be no air bubbles in the receiving chamber. Vertically press the GXSHW-EO-NLCs / PVA swelling micro-needle patch at the appropriate position on the rat skin, and fix it on the stratum corneum of the excised abdominal rat skin with medical adhesive tape after 1 min. During the treatment process, it is necessary to ensure the integrity of the skin. After the treatment, place the rat skin on the 7 mL receiving chamber, ensure that the stratum corneum of the skin faces the supply chamber, and place it between the supply chamber and the receiving chamber of the diffusion cell, and make sure that the skin is in full contact with the liquid level of the receiving solution. At the same time, set up another group. The skin of the rats in this group is not treated, and only the same amount of GXSHW-EO-NLCs solution is given. The effective area of the diffusion cell is 3.14 cm 2, the diffusion cell temperature was maintained at a constant (37 ± 0.5) °C, the speed of magnetic stirring was 300 rpm to simulate the internal circulation of blood and tissue fluid under the human skin, the sampling volume was 2 mL, and after sampling, the same volume of 30% ethanol normal saline at 37 °C treated by ultrasonic was supplemented. The sampling times were set at 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, 24 h, 48 h, and 72 h. After detection, the cumulative permeation amount per unit area of cinnamic acid at 24 h was 3.39 μg, and the cumulative transdermal rate was 40.67%; the cumulative permeation amount per unit area of costunolide at 24 h was 80.73 μg, and the cumulative transdermal rate was 57.19%, which proved that the GXSHW-EO-NLCs / PVA swelling microneedle patch could penetrate the skin and continuously and slowly release drugs in the skin tissue to exert the drug effect.

[0027] 5. Pharmacodynamic verification This experiment was carried out for the feeding and observation of experimental rats in the Experimental Animal Center of Shandong University of Traditional Chinese Medicine, and was approved by the Experimental Animal Ethics Committee of Shandong University of Traditional Chinese Medicine (approval number: SDUTCM20230511001). Sixty 6-week-old SPF-grade SD male rats were selected and provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. (license number: SCXK (Jing) 2021-0006). After numbering each rat, they were divided into a sham operation group, a model group, a GXSHW-EO-NLCs / PVA swelling microneedle application group, a blank microneedle matrix application group, a traditional acupoint application group, and a positive drug group by the random number table method, with 10 rats in each group. All rats had free access to water and food, the light-dark cycle was 12 / 12 h, the environmental temperature was maintained at 23 - 26 °C, and the humidity was maintained at (60 ± 10)%.

[0028] The treatment methods for each group were as follows.

[0029] Sham operation group: Thoracotomy was performed, and a suture was passed under the left anterior descending coronary artery, but no ligation was done. After successful modeling, the back of the rat was depilated and wiped clean, and the blank patch was applied to the bilateral Xinshu (BL15) and Jueyinshu (BL14) acupoints, with each application for 12 h and continuous intervention for 3 days.

[0030] Model group: Thoracotomy was performed, and the left anterior descending coronary artery was ligated. After successful modeling, the back of the rat was depilated and wiped clean, and the blank patch was applied to the bilateral Xinshu (BL15) and Jueyinshu (BL14) acupoints, with each application for 12 h and continuous intervention for 3 days.

[0031] Blank microneedle matrix application group: Thoracotomy was performed, and the left anterior descending coronary artery was ligated. After successful modeling, the back of the rat was depilated and wiped clean, and the blank microneedle matrix patch was applied to the bilateral Xinshu (BL15) and Jueyinshu (BL14) acupoints, with each application for 12 h and continuous intervention for 3 days.

[0032] GXSHW-EO-NLCs / PVA Swelling Microneedle Patch Group: Open the chest, ligate the left anterior descending coronary artery. After successful modeling, depilate and clean the back of the rats. Apply the GXSHW-EO-NLCs / PVA swelling microneedle patch on bilateral Xinshu (BL15) and Jueyinshu (BL14), apply for 12 hours each time, and intervene continuously for 3 days.

[0033] Traditional Acupoint Application Group: Open the chest, ligate the left anterior descending coronary artery. After successful modeling, depilate and clean the back of the rats. Grind Guanxin Suhe Pills and mix them with clear water into a paste, then apply the paste on bilateral Xinshu (BL15) and Jueyinshu (BL14), apply for 12 hours each time, and intervene continuously for 3 days.

[0034] Positive Drug Group: Open the chest, ligate the left anterior descending coronary artery. After successful modeling, intragastrically administer trimetazidine hydrochloride at a dose of 10 mg / (kg·d), and intervene continuously for 3 days.

[0035] After the intervention, when the rats are fully anesthetized, a specific person is responsible for weighing the rats and recording. Take blood from the abdominal aorta, take about 5 mL of blood. After completion, let it stand at room temperature for 30 minutes, centrifuge at 3000 rpm for 15 minutes at 4°C, carefully aspirate the upper serum and place it in a cryopreservation tube, quickly put it into liquid nitrogen, and then place it in a -80°C refrigerator for standby, which is used for the detection of myocardial enzymes and oxidative stress levels. When taking the hearts of the rats, quickly remove the rat hearts with scissors after opening the chest, wash them 3 times with pre-cooled 1X PBS solution. Randomly select 4 rats from each group, cut the left ventricles of the rats, place them in 4% paraformaldehyde, and fix them in a 4°C refrigerator overnight for tissue sectioning; cut the left ventricles of the remaining rat hearts in each group, place them in a cryopreservation tube, quickly put them into liquid nitrogen, and then transfer them to a -80°C refrigerator for freezing for subsequent experiments.

[0036] Determination of Heart Index: Take the rat hearts, cut off the large blood vessels and connective tissues, wash them 3 times with 1X PBS solution, dry them as much as possible with filter paper, and weigh them in a petri dish. Calculate the heart index of each group. The calculation formula is heart index = [(rat heart mass / rat body mass)] × 100%.

[0037] The increase in the heart index of rats is related to edema, primary necrosis of myocardial fibers, and increased intermuscular space caused by inflammation. The experimental results are shown in Table 1 below.

[0038] Table 1 Comparison of Heart Indexes of Experimental Rats in Each Group Note: Compared with the sham operation group, * P < 0.01, ** P < 0.05; compared with the model group, △ P < 0.01; compared with the GXSHW-EO-NLCs / PVA Swelling Microneedle Patch Group, ▽▽P <0.05; compared with the blank microneedle matrix application group, ○ P <0.01; compared with the traditional acupoint application group, □□ P <0.05.

[0039] It can be seen from the results analysis of Table 1 above that compared with the sham operation group, the cardiac index of the rats in the model group was significantly increased ( P <0.01), indicating that the myocardial interstitial space of the rats in the model group increased, which could reflect the successful establishment of the acute myocardial ischemia model. Compared with the model group, the cardiac indices of the rats in the GXSHW-EO-NLCs / PVA swelling microneedle application group, the blank microneedle matrix application group, the traditional acupoint application group, and the positive drug group were all decreased ( P <0.01). Compared with the GXSHW-EO-NLCs / PVA swelling microneedle application group, the cardiac index of the rats in the blank microneedle matrix application group increased ( P <0.05), and there was no significant difference in the cardiac indices of the rats in the traditional acupoint application group and the positive drug group ( P >0.05). Compared with the blank microneedle matrix application group, the cardiac index level of the rats in the positive drug group decreased ( P <0.01), and there was no statistical significance in the cardiac index level of the rats in the traditional acupoint application group ( P >0.05). Compared with the traditional acupoint application group, the cardiac index level of the rats in the positive drug group decreased ( P <0.05). In summary, after the intervention of each experimental protocol, the degree of cardiac injury of the rats in each group was improved to a certain extent. Acupoint application with the GXSHW-EO-NLCs / PVA swelling microneedle patch could significantly reduce the cardiac index of the rats.

[0040] The above description is only a preferred embodiment of the present invention, and it is not a limitation of the present invention in other forms. Any person skilled in the art may use the disclosed technical content to make changes or modifications into equivalent embodiments with equivalent changes and apply them to other fields. However, as long as it does not depart from the technical content of the present invention, any simple modification, equivalent change, and modification made to the above embodiments based on the technical essence of the present invention still belong to the protection scope of the technical solution of the present invention.

Claims

1. A preparation method of a volatile oil nano-liposome / PVA swelling type microneedle patch for Guanxin Suhe Pills, characterized in that, The steps are as follows: (1)Weigh the materials according to the following parts by weight: 45 - 55 parts of styrax, 100 - 110 parts of frankincense, 205 - 215 parts of sandalwood, and 205 - 215 parts of costus root. Crush them respectively, pass through a 100 - mesh sieve, and mix evenly to obtain mixture A. Add deionized water, seal and soak, then place it in a device with heating and condensation reflux, and keep it in a slightly boiling and condensation reflux state for at least 12 h. After standing, absorb the oil layer and add a desiccant to dry, to obtain the volatile oil of Guanxin Suhe Pills GXSHW - EO; (2)Add soybean lecithin, cholesterol, sodium cholate, and GXSHW - EO to a mixed solvent of methanol and chloroform, mix evenly and then perform rotary evaporation. After the rotary evaporation is completed, add a phosphate buffer solution, perform rotary evaporation again, then ultrasonicate and filter to obtain the volatile oil nano - lipidosomes of Guanxin Suhe Pills GXSHW - EO - NLCs solution; (3)Mix the PVA solution, PVP solution, citric acid solution, and GXSHW - EO - NLCs solution evenly, and centrifuge to obtain the microneedle solution. Pour the microneedle solution into a PDMS mold by vacuum casting method, evacuate, dry and demold to obtain the swelling microneedle patch.

2. The preparation method of the volatile oil nano-liposome / PVA swelling type microneedle patch of Guanxin Suhe Pills according to claim 1, characterized in that, In step (1), the mass of deionized water added is 5.8 - 6.2 times the mass of mixture A, the sealing and soaking time is 11 - 13 h, and the desiccant is anhydrous sodium sulfate.

3. The preparation method of the volatile oil nano-liposome / PVA swelling micro-needle patch of Guanxin Suhe Pills according to claim 1, characterized in that, In step (2), the mass ratio of soybean lecithin to GXSHW - EO is (3.9 - 4.1):1, the mass ratio of soybean lecithin to cholesterol is (5.9 - 6.1):1, the mass of sodium cholate is 2.0 - 3.0 mg; the volume ratio of methanol to chloroform in the mixed solvent is 1:1; the rotary evaporation speed is 150 - 170 rpm, the rotary evaporation temperature is 45 - 55 °C, and the pH of the phosphate buffer solution is 6.7 - 6.

9.

4. The preparation method of the volatile oil nano-liposome / PVA swelling type microneedle patch of Guanxin Suhe Pills according to claim 1, characterized in that, In step (2), the filtration is carried out using a 0.45 - μm microporous membrane.

5. The preparation method of the volatile oil nano-liposome / PVA swelling microneedle patch of Guixin Suhe Pills according to claim 1, characterized in that, In step (2), the mass fraction of the PVA solution is 3.5 - 4%, the mass fraction of the PVP solution is 5 - 5.5%, the mass fraction of the citric acid solution is 0.3 - 0.5%, and the mass fraction of the GXSHW - EO - NLCs solution is 15 - 20%; the volume ratio of the PVA solution:PVP solution:citric acid solution:GXSHW - EO - NLCs solution is (29 - 31):(33 - 36):(4 - 6):(29 - 31).

6. The preparation method of the volatile oil nano-liposome / PVA swelling type microneedle patch of Guixin Suhe Pills according to claim 1, characterized in that, In step (3), the bottom radius of the swelling microneedle patch is 120 - 123 μm, the height of the needle body is 500 - 502 μm; the angle of the microneedle tip is 29 - 32 µm, the tip spacing is 560 - 580 µm, and the number of needle bodies in each microneedle patch is 140 - 150.

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