Evodia rutaecarpa soluble microneedle for acupoint administration as well as preparation method and application thereof

Abstract

The invention belongs to the field of microneedle transdermal drug delivery, and particularly relates to an acupoint drug delivery soluble evodia rutaecarpa microneedle as well as a preparation method and application thereof in treatment of oral ulcer. The preparation method comprises the following steps: modifying effective biological components in fructus evodiae to obtain a solid dispersion, and then mixing the solid dispersion with a needle tip matrix to prepare the drug-loaded microneedle. Wherein the effective biological component in the fructus evodiae is evodiamine (EVO) or rutaecarpin (RUT). The medicinal evodia soluble drug-loaded microneedle is prepared, and the medicinal evodia soluble drug-loaded microneedle is attached to the pelma Yongquan acupoint to treat oral ulcer through matching of the medicinal evodia and the acupoint.

Description

Technical Field

[0001] The invention belongs to the field of microneedle transdermal drug delivery, and particularly relates to an evodia rutaecarpa soluble microneedle for acupoint drug administration, a preparation method thereof, and an application thereof in treating oral ulcers. Background Art

[0002] Oral ulcers are common oral diseases with unknown etiology, and it is difficult to develop countermeasures for their specific pathogenesis. When Western and Chinese medicines are administered systemically to treat oral ulcers, the intake of drugs will inevitably have other side effects on the body, and the efficacy of oral drugs is generally slow; and when Chinese and Western medicines are used for local treatment of symptoms, they have a certain degree of irritation to the wound, causing pain and foreign body sensation, and patient compliance is low. Therefore, it is hoped to seek other more effective ways of administration.

[0003] Applying Evodia rutaecarpa powder mixed with vinegar to the Yongquan acupoint under the foot is a drug administration method based entirely on the theoretical system of traditional Chinese medicine, and has been proven to be effective in clinical trials. However, its disadvantages are that the amount of Evodia rutaecarpa transdermal administration is extremely low, and its permeable components are greatly reduced compared to itself, resulting in high drug loss, and a large amount of drug residues remaining during use; applying powder to acupoints has low patient compliance, difficult drug administration, and inconvenience in daily life. At present, the components that can be specifically and clearly measured to have a certain amount of permeability are evodiamine and evodiamine, so the acupoint application drugs are improved based on the theoretical system of traditional Chinese medicine.

[0004] As the largest organ of the human body, the skin is a powerful barrier that protects us from physical, chemical, biological and other harms from the outside world. As a protective barrier between the body and the outside world, the skin protects the body from allergens, pathogens and other organisms from the outside to the inside, and also ensures that the body's water will not be lost from the inside to the outside. But at the same time, the skin does have a certain ability to absorb various substances. The skin can be divided into the stratum corneum, epidermis, dermis and subcutaneous tissue, and the barrier function of the stratum corneum of the skin limits the vast majority of drugs, generally through small molecule drugs and lipophilic drugs, and macromolecular drugs are difficult to be administered through the skin, which is the main obstacle to transdermal administration. Therefore, transdermal administration generally increases the amount of drug permeation, similar to adding a permeation enhancer to achieve the effect of permeation enhancement, or changing the drug preparation to prepare liposomes, ethosomes, vesicles, etc. to increase the amount of permeation, or using physical permeation enhancement, ultrasound, electroporation, etc. to increase the amount of drug permeation.

[0005] Microneedles are composed of dozens or hundreds of micron-sized needles that are neatly arranged to form an array. The length of the microneedle tip ranges from 25 to 1500 μm. This length range has the ability to penetrate the stratum corneum to release drugs without reaching the subcutaneous muscle tissue and other parts. Microneedles can bypass the first-pass effect and intestinal metabolism. In the case of transdermal administration, they avoid the above three common methods and open up a channel to penetrate the stratum corneum through the microneedle tip to avoid the stratum corneum barrier. The drug passes through the stratum corneum of the skin and reaches the designated part, where it is absorbed by the capillaries to achieve the drug administration effect. During the use of microneedles, hard needles can cause pain and inflammation, so soluble microneedles are developed, but there are certain technical difficulties in mass production of soluble microneedles. Summary of the invention

[0006] The present invention aims to provide an improved dosage form of acupoint application medicine based on the theoretical system of traditional Chinese medicine, and to provide a soluble microneedle of Evodia rutaecarpa for acupoint administration, a preparation method thereof, and an application thereof in treating oral ulcers.

[0007] To achieve the above object, the object of the present invention is achieved through the following technical solutions:

[0008] A method for preparing soluble microneedles of Evodia rutaecarpa for acupoint drug administration, comprising modifying effective biological components in Evodia rutaecarpa to obtain a solid dispersion, and then mixing the solid dispersion with a blank needle tip matrix to prepare drug-loaded microneedles; wherein the effective biological components in Evodia rutaecarpa are evodiamine (EVO) or rutaecarpine (RUT), and the structural formulas are respectively

[0009]

[0010] The solid dispersion is prepared by adding evodiamine (EVO) or rutaecarpine (RUT) into a solvent and stirring until completely dissolved. After dissolution, polyvinyl pyrrolidone (PVP K30) is added to the system under stirring conditions, and then rotary evaporation is performed to obtain a viscous product, which is then dried and dispersed to obtain a solid dispersion.

[0011] The solid dispersion is prepared by adding evodiamine (EVO) or rutaecarpine (RUT) into a solvent, respectively, and magnetically stirring for 60 minutes in a 60°C water bath until the drug is completely dissolved in the solvent. PVP K30 is then added and magnetically stirred for 2 hours. The solution is rotary evaporated, and ethanol is recovered under reduced pressure until it becomes viscous. The solution is then transferred to a 60°C vacuum drying oven and dried for 12 hours. The solid is taken out, ground into powder using a mortar, and passed through a 100-mesh sieve to obtain a solid dispersion.

[0012] The evodiamine (EVO) and the solvent are mixed at a ratio of 1:5 (w / v); the rutaecarpine (RUT) and the solvent are mixed at a ratio of 2:5 (w / v); the evodiamine (EVO) and polyvinyl pyrrolidone (PVP K30) are mixed at a ratio of (1:1-8; w / w); and the rutaecarpine (RUT) and polyvinyl pyrrolidone (PVP K30) are mixed at a ratio of (1:1-8; w / w).

[0013] The solvent for preparing the solid dispersion is anhydrous ethanol.

[0014] The solid dispersion, needle tip matrix and water are mixed to form a needle tip solution, wherein the solid dispersion and the blank needle tip matrix solution are mixed in a mass to volume ratio (20-40:1; w / v); the blank needle tip matrix is ​​selected from a combination of two or more of polyvinyl pyrrolidone (PVP K30), enzymatically cleaved oligomeric sodium hyaluronate (HA), and chondroitin sulfate (CS).

[0015] Preferably, the needle tip matrix is ​​one of PVP / CS (PVP accounts for 20% of the system concentration / CS accounts for 5-10% of the system concentration, w / w), PVP / HA (PVP accounts for 20% of the system concentration / HA accounts for 5-10% of the system concentration, w / w) or PVP / HA (PVP accounts for 5-15% of the system concentration / HA accounts for 15-5% of the system concentration, w / w).

[0016] More preferably, it is PVP / HA (PVP accounts for 5-15% of the system concentration / HA accounts for 15-5% of the system concentration, w / w).

[0017] The microneedles were prepared by centrifugation. A needle tip solution was added to a microneedle mold, and the tube was centrifuged at 3000 rpm for 5 minutes and rotated 90 degrees for centrifugation. This process was repeated 4 times to allow the centrifuge tube to rotate a full circle. The tube was dried at 25°C for 30 minutes, a backing matrix was added, and the tube was centrifuged for 5 minutes. The tube was rotated 180 degrees and centrifuged for another 5 minutes. The backing matrix aqueous solution was then added, and the tube was placed in a vacuum drying oven, dried at 25°C for 24 hours, and taken out after solidification.

[0018] The volume ratio of the needle tip solution to the backing matrix solution is 1:5 (V / V).

[0019] When preparing microneedles by the centrifugal method, the microneedles are placed in a centrifuge and injection molded by centrifugation. However, when the centrifuge speed is increased or decreased, it is difficult to place the centrifuge tube in a horizontal state. When the centrifuge tube is tilted, the mold also has a certain tilt. Due to the centrifugal effect and gravity, it is difficult for the matrix in the mold to be horizontally laid on the microneedle body. Therefore, in the preparation process, the centrifugal process is optimized, and the mold is rotated and centrifuged in a direction of 90° until a full rotation.

[0020] The microneedle curing method is as follows: curing is performed in a vacuum drying oven at a curing temperature of 25°C.

[0021] A kind of Evodia rutaecarpa soluble microneedle for acupoint administration prepared by the method, the Evodia rutaecarpa soluble microneedle for acupoint administration prepared by the method is non-irritating to human body, can be absorbed by human body, has high administration efficiency and high compliance.

[0022] An application of the Evodia rutaecarpa soluble microneedle for acupoint administration, and an application of the Evodia rutaecarpa soluble microneedle for acupoint administration in the preparation of a patch medicine for treating oral ulcers.

[0023] The administration method of the application is acupoint administration, and the acupoint for administration is Yongquan acupoint.

[0024] Compared with the prior art, the Evodia rutaecarpa soluble microneedle and its preparation method and application for treating oral ulcers of the present invention have the following beneficial effects:

[0025] 1. Based on the improved dosage form of acupuncture point patch drugs under the theoretical system of traditional Chinese medicine, the soluble drug-loaded microneedles have a higher efficacy in transdermal penetration than the patch. Compared with the patch, microneedle drug delivery can penetrate the stratum corneum. The drug can be delivered as continuously as possible, and the drug is prepared into a solid dispersion, and its intradermal retention is increased. The part of the drug retained in the skin can still be continuously delivered for a certain period of time in the future, taking into account the intradermal retention of microneedle acupuncture points and continuous delivery.

[0026] 2. The Evodia rutaecarpa soluble microneedles prepared by the preferred formula of the present invention include a main drug, an excipient and a matrix. The main drug (evodiamine, evodiamine alkaloids), excipients (polyvinyl pyrrolidone PVP K30) and a matrix (polyvinyl pyrrolidone PVP K30, polyvinyl alcohol PVA, enzyme-cut oligomeric sodium hyaluronate HA) are non-irritating to the human body, can be absorbed by the human body, and can be administered through microneedles through the skin. The skin can heal itself in a short time, and it is easy to stick and remove without residue.

[0027] 3. The present invention uses simple and effective materials, uses the effective ingredient of Evodia rutaecarpa, and enhances the transdermal absorption of the effective ingredient through degree stimulation and microneedle administration. The medicine acupoints are matched to achieve the effects of guiding fire downward, warming the meridians and dispersing cold, regulating qi and relieving pain, and have a therapeutic effect on oral ulcers.

[0028] 4. The matrix material used in the Evodia rutaecarpa soluble microneedles of the present invention is a material that can be absorbed by the human body, and the drug and the matrix have good miscibility. No organic solvent is required in the production process of the Evodia rutaecarpa soluble microneedles, and the production process is safe, energy-saving, and environmentally friendly. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is the cumulative dissolution curve of evodiamine (n=3).

[0030] Figure 2 This is the cumulative dissolution curve of Evodia rutaecarpine (n=3).

[0031] Figure 3 Microneedles were prepared in one step with different matrix ratios.

[0032] Figure 4 Morphological images of microneedles fabricated for single matrices at different scales.

[0033] Figure 5 The morphology of microneedle tips prepared with different matrix ratios.

[0034] Figure 6 For microneedle penetration M membrane picture.

[0035] Figure 7 Morphological images of microneedle backings prepared with different matrix ratios.

[0036] Figure 8 In vitro release of drug-loaded microneedles in low and high dose groups, including EVO cumulative release rate curve (A), RUT cumulative release rate curve (B), EVO cumulative release amount curve (C) and RUT cumulative release amount curve (D) (n=3).

[0037] Fig. 9 Transdermal release of drug-loaded microneedles in low-dose and high-dose groups Transdermal release of EVO low-dose and high-dose groups (A), transdermal release of RUT low-dose and high-dose groups (B) (n=3).

[0038] Fig.10 Transdermal release of drug-loaded microneedles in low-dose group and peeling group, including transdermal release of EVO low-dose group and peeling group (A), transdermal release of RUT low-dose group and peeling group (B) (n=3). DETAILED DESCRIPTION

[0039] The present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments. Unless otherwise specified, the experimental methods used below are all conventional methods in the art, and the ingredients or materials used are all ingredients or materials available through commercial channels unless otherwise specified. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments.

[0040] The present invention is based on the improved dosage form of acupoint application medicine under the theoretical system of traditional Chinese medicine, applies Evodia rutaecarpa to Yongquan acupoint for treating oral ulcers, and improves the preparation into Evodia rutaecarpa soluble microneedle acupoint administration for administration through acupoint stimulation. By improving the dissolution of biological effective components in Evodia rutaecarpa, screening the preparation method, preparation process, preparation matrix type, ratio, and concentration of soluble microneedles, optimizing the preparation process, combining the effective components with improved dissolution with the screened microneedle matrix, and preparing Evodia rutaecarpa soluble drug-loaded microneedles.

[0041] Example 1

[0042] The preparation method of the solid dispersion of the effective ingredient of Evodia rutaecarpa in this example is:

[0043] (1) Accurately weigh 200 mg of EVO API and dissolve it in 1000 mL of anhydrous ethanol. Stir magnetically in a 60°C water bath for 60 min until the drug is completely dissolved in the solvent. Add an appropriate amount of PVP K30 (200 mg, 400 mg, 800 mg, 1200 mg, 1600 mg) and continue magnetic stirring for 2 h. Then, perform rotary evaporation on the solution, recover ethanol under reduced pressure until it becomes viscous, transfer it to a 60°C vacuum drying oven, dry it for 12 h, take out the solid, grind it in a mortar, and pass it through a 100-mesh sieve to obtain EVO-PVP SD (1:1, 1:2, 1:4, 1:6, 1:8) of various ratios.

[0044] (2) Accurately weigh 400 mg of RUT raw material and dissolve it in 1000 mL of anhydrous ethanol. Stir magnetically in a 60°C water bath for 60 min until the drug is completely dissolved in the solvent. Add an appropriate amount of PVP K30 (400 mg, 800 mg, 1600 mg, 2400 mg, 3200 mg) and continue magnetic stirring for 2 h. Then, perform rotary evaporation on the solution, recover ethanol under reduced pressure until it becomes viscous, transfer it to a 60°C vacuum drying oven, dry it for 12 h, take out the solid, grind it in a mortar, and pass it through a 100-mesh sieve to obtain RUT-PVP SD (1:1, 1:2, 1:4, 1:6, 1:8) of various ratios.

[0045] Based on this example, the effects of different raw material and auxiliary material ratios on the cumulative dissolution of evodiamine and evodiamine were investigated. Figure 1 and Figure 2 .

[0046] Depend on Figure 1 and Figure 2It can be seen that among the dissolution of EVO, RUT raw materials, EVO, RUT physical mixtures, and various proportions of EVO-PVP SD and RUT-PVP SD, the dissolution rate of EVO-PVP SD (EVO: PVP K30, 1:2, w / w) is faster than that of other proportions, and the solubility is the best, with the dissolution rate reaching more than 90% within 6 hours; the solid dispersions of various proportions of RUT (except RUT: PVP K30, 1:1, w / w), the other dissolution rates can reach 80% within 24 hours. Considering the subsequent preparation of microneedle preparations, the preparation ratio with less excipients is selected to increase its drug loading capacity, so EVO-PVP SD (EVO: PVP K30, 1:2, w / w); RUT-PVP SD (RUT: PVP K30, 1:2, w / w) are preferred.

[0047] Example 2

[0048] The injection molding method of the soluble microneedle generally adopts a casting method. There are three common casting methods. After the drug and the matrix are mixed, they are mixed with water and stirred to swell, and then cast on the microneedle mold by high-speed centrifugation, vacuum method and ultrasonic vibration method. Different injection molding methods have different effects on the shaping of the microneedle. The injection molding methods for preparing microneedles are screened.

[0049] The preparation method adopted by the injection molding method for preparing microneedles in this example is:

[0050] (1) A one-step microneedle method was used, that is, the microneedle matrix was directly prepared into microneedles through different injection molding methods. PVP K30 was selected as the matrix, and the mixing ratio was PVP K30: water (3:10, w / v). It was used as a blank matrix solution for screening the injection molding method.

[0051] (2) One-step preparation of microneedles using high-speed centrifugation: 100 μL of blank matrix solution was injected into the mold, centrifuged at 3000 rpm for 5 min, the centrifuge tube was rotated 180° and centrifuged again, and the above method was repeated twice. Then 200 μL of matrix was added and solidified.

[0052] (3) Vacuum method for preparing microneedles in one step: 300 μL of blank matrix solution was injected into the mold. The mold was placed in a vacuum drying oven at 25°C and vacuumed under negative pressure. After 24 hours of curing, the mold was taken out.

[0053] (4) Ultrasonic method for preparing microneedles in one step: 300 μL of blank matrix solution was injected into the mold. The mold was placed in the instrument for ultrasonic treatment for 20 min and then taken out to wait for the microneedles to be formed. The test results are shown in Table 1.

[0054] Table 1 Evaluation of microneedles prepared by different methods

[0055]

[0056] The high-speed centrifugation method has a high needle formation rate and good shaping. In the vacuum method, the viscosity of the matrix is ​​too high, and it is difficult to completely inject the matrix into the mold by vacuuming once, and repeated vacuuming takes a long time, and there are still bubbles remaining. In the ultrasonic injection molding method, the ultrasonic power is low, the matrix is ​​sticky and the matrix adheres to the surface of the mold, the matrix is ​​not injected into the mold, and the matrix does not enter the needle tip of the mold body, which is mostly bubbles, and ultrasonic bubbles are difficult to remove. The embodiment preferably uses the high-speed centrifugation method to prepare soluble microneedles.

[0057] Example 3

[0058] The curing method of the soluble microneedles, the different curing conditions, temperatures and environments of the microneedles have a direct impact on the shaping of the microneedles. The preparation and curing methods of the microneedles (ie, blank matrices) prepared by different methods are screened.

[0059] The microneedles were prepared under the temperature conditions of 25°C, 35°C, and 60°C, and under the environmental conditions of oven drying, room temperature drying, and vacuum drying oven drying. The results are shown in Table 2.

[0060] Table 2 Evaluation of different curing methods on microneedle forming

[0061]

[0062] When the curing temperature for microneedle preparation is selected between 35 and 60°C, the temperature is too high, the moisture on the surface of the patch is lost first, and the loss is too fast, so that the needle body and the backing part cannot be cured evenly, the backing part loses water severely and shrinks, or the patch warps up around it, and the microneedle tip is easy to break.

[0063] At a curing temperature of 25°C, the backing surface is slightly uneven when dried in a forced air oven. The humidity of the room temperature drying environment has a certain influence, and the curing time is slightly longer.

[0064] The vacuum drying oven is used for drying and curing, and the needle rate is good, the backing is flat and easy to demould, and it is preferred to use a vacuum drying oven for curing.

[0065] Example 4

[0066] The matrix was selected and fused with water, and after swelling, the microneedles were prepared by high-speed centrifugation in one step. A single matrix material for microneedle preparation was selected for preparation, including hyaluronic acid (HA), polyvinyl pyrrolidone (PVP), polyvinyl alcohol (PVA) and chondroitin sulfate (CS). Different proportions of matrix PVP (10%, 20%, 30%), HA (10%, 20%, 30%), CS (10%, 20%, 30%), PVA (5%, 10%, 15%) were selected to prepare blank microneedles.

[0067] The preparation method of the blank soluble microneedle is:

[0068] (1) The high-speed centrifugation method in the one-step preparation of microneedles: 100 μL of blank single matrix solution was injected into the mold, centrifuged at 3000 rpm for 5 min, the centrifuge tube was rotated 180° and centrifuged again, and the above method was repeated twice. After adding 200 μL of matrix, it was placed in a vacuum drying oven and dried at 25°C for 24 h. After solidification, it was taken out.

[0069] (2) The blank single matrix solution, water and needle tip matrix are mixed. Matrix hyaluronic acid (HA), polyvinyl pyrrolidone (PVP), polyvinyl alcohol (PVA) and chondroitin sulfate (CS) are selected. Different proportions of matrix PVP (PVP accounts for 10-30% of the system concentration), HA (HA accounts for 10-30% of the system concentration), CS (CS accounts for 10-30% of the system concentration), PVA (PVA accounts for 5-15% of the system concentration) are selected to prepare blank microneedles.

[0070] The microneedles were screened based on their overall appearance and the condition of the needle tips observed under a microscope. Figure 3 , 4 .

[0071] From the overall morphology, in a single matrix, regardless of the proportion of CS, its material properties are not suitable for microneedle shaping. PVP and HA with a 10% ratio have a large area of ​​missing needles after demolding due to the low matrix content. It can be seen that this phenomenon will occur at low matrix concentrations. After curing, PVA has shrinkage at different proportions, and due to its high viscosity, it is difficult to exhaust all bubbles.

[0072] Except for the situation where a large area of ​​needles are missing due to a slightly lower matrix content, all matrices can basically form microneedle tips. In terms of shaping, it can be seen that the PVA needle tip is slightly bent. Because of its soft texture, it is difficult to keep the needle tip sharp during demolding. In addition, it should be considered whether other situations will be encountered during use.

[0073] As shown in the figure, the appearance and needle formation rate of a single matrix in the process of preparing microneedles are different. The overall appearance inspection is listed in the figure. In addition to the phenomenon of missing needles, the overall appearance such as shrinkage and mechanical strength should also be inspected. As shown in Table 3

[0074] Table 3 Appearance observation of blank microneedles prepared by single matrix one-step method

[0075]

[0076] In the process of preparing microneedles using a single matrix, there are different defects in the appearance morphology and needle formation rate.

[0077] First, in terms of appearance, PVP 10% has less matrix, so most of the needle tip part breaks during demoulding. During the curing process, the water loss is extremely large, making it difficult to form a patch, and its backing part is hollowed out over a large area. PVP 20% also has less matrix and can form a complete patch, but its backing part is thinner, and when the microneedle is pressed slightly, the backing breaks. PVP 30% has no obvious defects in appearance and morphology, and performs well in needle formation rate, but its backing still lacks toughness, and it will break when pressed hard. In subsequent experiments, it was found that the material of the single PVP matrix for preparing microneedles has a certain hygroscopic dissolution phenomenon. The longer it is placed at room temperature, the more obvious the phenomenon is, until the needle tip disappears completely and the backing is not formed. PVP does not have the conditions for preparing microneedles with a single matrix.

[0078] HA 10% matrix is ​​less after solidification, accompanied by water loss, and it is difficult to form a complete patch. Because there is no backing force, the needle body breaks in the mold in a large area during demoulding, and it does not form a patch. HA 20% matrix increases slightly, but hyaluronic acid is very brittle and has no toughness after being used as a microneedle. The patch will break with a little force. HA 30% can form a complete patch with the increase of matrix, but due to the nature of the material, the patch is severely brittle and cannot be pressed. HA does not have the conditions for preparing microneedles with a single matrix.

[0079] All proportions of CS are difficult to form into patches. After CS is mixed with water, molded and cured, it has no plastic properties. After drying, it forms blocky particles, which are spread flat on the bottom of the mold like gravel. It falls apart when touched and is difficult to form. CS does not have the conditions for preparing microneedles with a single matrix.

[0080] PVA (5%, 10%, 15%) matrices in various proportions can be molded into microneedle patches, but due to the properties of the material, PVA solidifies after swelling, and there are no defects in the appearance morphology. However, the matrix itself is soft, and the needle tip becomes soft when pressed with force, and has no puncture force. Its mechanical strength is far from enough to penetrate the skin. PVA does not have the conditions for preparing microneedles as a single matrix.

[0081] A single matrix does not have the conditions for preparing microneedles. During the preparation process, the characteristics of each material matrix are gradually understood to ensure that excellent conditions are screened out. It is preferred to mix the matrix and prepare microneedles in two steps.

[0082] Example 5

[0083] According to the different properties required of the microneedle tip and the backing, the tip needs to have the mechanical strength to penetrate the skin, and the backing needs to withstand a certain pressure and have a little toughness. The microneedles are prepared by a two-step method.

[0084] The microneedles are prepared by a two-step method, in which the needle tip and the backing part are separated and prepared using different matrices, so that on the same patch, the needle tip is sharp, has strong puncture force, and the backing can withstand pressure and has toughness.

[0085] The preparation method of the blank soluble microneedle is:

[0086] (1) Preparation of microneedles by two steps using high-speed centrifugation method: 50 μL of needle tip matrix solution was added to the microneedle mold, centrifuged at 3000 rpm for 5 min, centrifuged three times, dried at 25°C for 30 min, 100 μL of backing matrix aqueous solution was added, centrifuged for 5 min, and then 150 μL of backing matrix was added, placed in a vacuum drying oven, and dried at 25°C for 24 h. After solidification, it was taken out.

[0087] (2) The backing concentration was fixed at PVP / PVA (20% / 5%, w / w) and microneedles were prepared by a two-step centrifugation method.

[0088] The above-mentioned needle tip solution is a mixture of water and needle tip matrix, and the matrix PVP / CS is selected (PVP accounts for 20% of the concentration in the system / CS accounts for 5% of the concentration in the system, w / w), PVP / CS (PVP accounts for 20% of the concentration in the system / CS accounts for 10% of the concentration in the system, w / w), PVP / HA (PVP accounts for 20% of the concentration in the system / HA accounts for 5% of the concentration in the system, w / w), PVP / HA (PVP accounts for 5% of the concentration in the system / HA accounts for 10% of the concentration in the system, w / w), PVP / HA (PVP accounts for 5% of the concentration in the system / HA accounts for 15% of the concentration in the system, w / w), PVP / HA (PVP accounts for 10% of the concentration in the system / HA accounts for 10% of the concentration in the system, w / w), PVP / HA (PVP accounts for 10% of the concentration in the system / HA accounts for 10% of the concentration in the system, w / w), PVP / HA (PVP accounts for 15% of the concentration in the system / HA accounts for 5% of the concentration in the system, w / w).

[0089] The appearance of the microneedles was observed under a microscope. M (sealing film) is used as the evaluation standard for examining mechanical strength. The microneedle is pressed vertically on the 5 layers M, press for 2 minutes and then remove it to observe the penetration of microneedles M's situation and puncture rate to verify its mechanical strength. Use a microscope to observe the morphology of the microneedle: see Figure 5

[0090] For the prepared soluble microneedles, after the needle tip solution containing chondroitin sulfate (CS) is prepared into a microneedle patch, the chondroitin sulfate (CS) component therein is dried due to the curing process, and the gritty feeling after drying is restored. A part of it floats on the surface of the backing, making it difficult to demold, and a part of it remains in the needle tip, causing the needle tip to break; PVP / HA (20% / 5%, w / w) and PVP / HA (20% / 10%, w / w) as needle tip matrices are basically intact in appearance, but because the content of PVP is relatively large, when the content of PVP is high, bubbles are easily retained in the patch during the molding process; under the premise of a certain total amount, the volume of HA is small. The overall needle tip matrix of PVP / HA (5% / 15%) is smaller in weight, and the solid content of the matrix is ​​less. Its mechanical strength is slightly weaker than the other two groups. In addition, during the preparation process, the needle tip part solidifies first, but the solid content is low, and the insufficient needle tip part will be used to fill the backing part. During this process, cavitation is easily formed in the backing part. PVP / HA (15% / 5%) and PVP / HA (10% / 10%) are similar. Considering the influence of PVP dissolution, PVP / HA (10% / 10%) is preferred.

[0091] Example 5 In the mechanical strength test, the penetration M evaluates mechanical strength. Generally, after penetrating the third layer of membrane, the microneedle will leave a mark of pressure on the fourth layer of membrane, but will not penetrate the fourth layer of membrane. See Table 4. Figure 6 .

[0092] Table 4 Penetration rate of microneedle tips prepared with different matrix ratios

[0093]

[0094] According to the appearance and mechanical strength, the preferred microneedle tip matrix is ​​PVP / HA (10% / 10%).

[0095] Example 6

[0096] A kind of Evodia rutaecarpa soluble drug-loaded microneedle, the microneedle backing needs to have a certain hardness to enable the needle tip to puncture when a certain pressure is applied, and has a little toughness so that the backing will not be broken due to force during the process of applying pressure.

[0097] The preparation method of the blank soluble microneedle is:

[0098] (1) Preparation of microneedles by two steps using high-speed centrifugation: 50 μL of blank needle tip matrix solution was added to the microneedle mold, centrifuged at 3000 rpm for 5 min, centrifuged three times, dried at 25°C for 30 min, 100 μL of backing matrix aqueous solution was added, centrifuged for 5 min, and then 150 μL of backing matrix was added, placed in a vacuum drying oven, and dried at 25°C for 24 h. After solidification, the mold was taken out.

[0099] (2) The preparation method uses PVP / HA (10% / 10%) as the needle tip matrix.

[0100] (3) The backing matrix selected PVP / PVA (PVP accounts for 10% of the system concentration, PVA accounts for 5% of the system concentration), PVP / PVA (PVP accounts for 10% of the system concentration / PVA accounts for 10% of the system concentration), PVP / PVA (PVP accounts for 20% of the system concentration / PVA accounts for 5% of the system concentration), PVP / PVA (PVP accounts for 20% of the system concentration / PVA accounts for 10% of the system concentration) to investigate the morphology and usability of different proportions. Figure 7 .

[0101] For the prepared soluble microneedles, PVP / PVA (10% / 5%) (10% / 10%) (20% / 5%) (20% / 10%) all have a complete appearance and form a base, PVP / PVA (10% / 5%), PVP / PVA (10% / 10%) contain less solid matter, the backing is thinner, and liquid needs to be added during preparation. The backing is softer, and the backing is difficult to withstand the pressure when the microneedle is pressed hard, making it difficult for the needle body to puncture the skin.

[0102] When preparing the backing with PVP / PVA (20% / 10%), the PVP content is relatively large and the backing matrix is ​​relatively sticky. When pressed, the needle tip is subjected to a large mechanical strength, the needle tip shrinks into the backing, and the penetrating force is weakened.

[0103] The backing is preferably PVP / PVA (20% / 5%), so that the backing can withstand pressure and has a certain toughness and will not break under stress.

[0104] Example 7

[0105] A method for preparing soluble drug-loaded microneedles of Evodia rutaecarpa, an improved dosage form of acupoint application medicine based on the theoretical system of traditional Chinese medicine.

[0106] Optimization of microneedle centrifugation preparation process:

[0107] During the speed increase and decrease process of the centrifuge, the centrifuge tube turns from horizontal to tilted. During this process, the liquid in the microneedle mold tilts due to the centrifugal force, making it difficult for the needle tip solution to be completely spread in the mold. The process is optimized to rotate the mold 90° and continue centrifugation for 5 minutes after each centrifugation for 5 minutes during the needle tip injection molding process. This is repeated 4 times to make the mold rotate a full circle. After the needle tip is solidified, the backing part will also be affected by the centrifugal tilting force. Therefore, the centrifugation of the backing part is optimized from the original 5 minutes of centrifugation to 5 minutes of centrifugation, then rotate 180° and centrifuge for another 5 minutes.

[0108] The optimized process is as follows: add 50 μL of needle tip matrix solution to the microneedle mold, centrifuge at 3000 rpm for 5 minutes, rotate the centrifuge tube 90 degrees and continue centrifugation, repeat this process 4 times, and make the mold rotate a full circle. Dry at 25°C for 30 minutes, add 100 μL of backing matrix solution, centrifuge for 5 minutes, rotate the centrifuge tube 180 degrees and centrifuge for another 5 minutes. Then add 150 μL of backing matrix solution, place in a vacuum drying oven, and dry at 25°C for 24 hours. Take out after curing.

[0109] Example 8

[0110] A method for preparing an improved dosage form of Evodia rutaecarpa soluble drug-loaded microneedle for acupoint application based on the theoretical system of traditional Chinese medicine. According to Examples 1 to 7, the Evodia rutaecarpa soluble drug-loaded microneedle is prepared.

[0111] The preparation method used in this example is:

[0112] (1) Accurately weigh 200 mg of EVO API and dissolve it in 1000 mL of anhydrous ethanol. Stir magnetically in a 60°C water bath for 60 min until the drug is completely dissolved in the solvent. Add 400 mg of PVP K30 and continue to stir magnetically for 2 h. Then, perform rotary evaporation on the solution, recover ethanol under reduced pressure until it becomes viscous, transfer it to a 60°C vacuum drying oven, dry it for 12 h, take out the solid, grind it in a mortar, and pass it through a 100-mesh sieve to obtain EVO-PVP SD in a 1:2 ratio.

[0113] (2) Accurately weigh 400 mg of RUT raw material and dissolve it in 1000 mL of anhydrous ethanol. Stir magnetically in a 60°C water bath for 60 min until the drug is completely dissolved in the solvent. Then add 800 mg of PVP K30 and continue to stir magnetically for 2 h. Then, perform rotary evaporation on the solution, recover ethanol under reduced pressure until it becomes viscous, transfer it to a 60°C vacuum drying oven, dry it for 12 h, take out the solid, grind it in a mortar, and pass it through a 100-mesh sieve to obtain RUT-PVP SD in a 1:2 ratio.

[0114] (3) Selecting and mixing the prepared solid dispersions of evodiamine and evodiamine according to a mass ratio of evodiamine: evodiamine (7:3; w / w) to form a solid dispersion mixture of evodiamine and evodiamine.

[0115] (4) The prepared solid dispersion mixture of evodiamine and evodiamine was mixed with the needle tip matrix solution (5:1, w / v), and after vortexing for 3 minutes, a drug-loaded needle tip solution was prepared, and microneedles were immediately prepared.

[0116] (5) The needle tip matrix solution is prepared by mixing polyvinyl pyrrolidone (PVP K30) and enzymatic oligomeric hyaluronate (HA) with water, and the two are mixed in a mass ratio of PVP / HA (PVP accounts for 10% of the system concentration / HA accounts for 10% of the system concentration, w / w). The backing matrix solution is prepared by mixing polyvinyl pyrrolidone (PVP K30) and polyvinyl alcohol (PVA) with water, and the two are mixed in a mass ratio of PVP / PVA (PVP accounts for 20% of the system concentration / PVA accounts for 5% of the system concentration).

[0117] (6) Preparation by centrifugation, optimization of microneedle centrifugation preparation: add 50 μL of drug-loaded needle tip solution to the microneedle mold, centrifuge at 3000 rpm for 5 min, rotate 90° for centrifugation, repeat this 4 times, and make the mold rotate a full circle. Dry at 25°C for 30 min, add 100 μL of backing matrix solution, centrifuge for 5 min, rotate 180° and centrifuge for another 5 min, then add 150 μL of backing matrix solution, place the mold in a vacuum drying oven, and dry at 25°C for 24 hours. After curing, take out the microneedle patch to obtain the soluble drug-loaded microneedles of Evodia rutaecarpa.

[0118] Example 9

[0119] An improved dosage form of Evodia rutaecarpa soluble drug-loaded microneedles for acupoint application based on the theoretical system of traditional Chinese medicine, and an in vitro release test.

[0120] According to Example 8, two groups of low-dose and high-dose microneedles were prepared: EVO-PVP SD: RUT-PVP SD (low-dose group: 700μg: 300μg; high-dose group: 1400μg: 600μg), and the microneedles were placed in a conical flask containing 50mL of ethanol-PEG400-normal saline (4:3:3), and a water bath was selected at 37°C with magnetic stirring. Samples were taken at time points (5min, 10min, 15min, 30min, 45min, 60min), and 1mL was taken at each pre-set time point, and passed through a 0.22μm filter membrane to enter the liquid phase, and the liquid was replenished at the same time. The cumulative release rate and cumulative release amount of the two drugs were calculated. See Figure 8 .

[0121] The cumulative release rate of EVO in the low-dose drug-loaded microneedle group was 97.8±3.4% at 30 minutes, and the cumulative release rate of RUT at 30 minutes was 98.0±3.3%; the cumulative release rate of EVO in the high-dose drug-loaded microneedle group was 96.2±2.4% at 30 minutes, and the cumulative release rate of RUT at 30 minutes was 99.4±1.5%. After the soluble microneedle preparation of Evodia rutaecarpa was formed, its active ingredients could still achieve a high release amount, and the dosage form of the preparation did not affect the drug release.

[0122] Example 10

[0123] An improved dosage form of acupoint application drug based on the theoretical system of traditional Chinese medicine, Evodia rutaecarpa soluble drug-loaded microneedles, in vitro transdermal test.

[0124] The ex vivo mouse skin of SD rats was taken and the excess hair was removed. The microneedles prepared according to Example 7 were divided into two groups: EVO-PVP SD and RUT-PVP SD (low-dose group: 700 μg: 300 μg; high-dose group: 1400 μg: 600 μg). The drug-loaded microneedles were pressed on the ex vivo mouse skin for 5 min each, and the mouse skin with microneedles inserted was fixed between the drug administration pool and the receiving pool of the vertical transdermal instrument (the effective receiving area in the receiving pool is 0.68 cm 2 ). Inject 4 mL of ethanol-PEG400-normal saline (4:3:3) into the receiving pool. Keep the temperature close to the skin temperature at 32°C and the magnetic stirring at 600 r·min-1. The sampling time points are (2h, 4h, 6h, 8h, 10h, 12h, 18h, 24h, 48h), take 1.5 mL of solution at each sampling point, pass through a 0.22 filter membrane and wait for the liquid phase to enter, and replenish the liquid. When the vertical transdermal instrument is in operation, the part where the microneedle contacts the skin is prone to carry tiny bubbles. After injecting the liquid into the receiving pool, pay attention to exhausting the bubbles from the part in contact with the skin, and after the receiving pool is magnetically stirred for 10 minutes, observe the receiving pool again for tiny bubbles and exhaust them to prevent gas from interfering with the test data.

[0125] The efficacy of microneedles in transdermal penetration is higher than that of patches. In the transdermal penetration part, the low-dose group EVO had a penetration of 20.65±4.35μg / cm2 within 48h; the high-dose group EVO had a penetration of 35.65±11.33μg / cm2 within 48h; the low-dose group RUT had a penetration of 20.69±3.94μg / cm2 within 48h; the high-dose group RUT had a penetration of 35.06±8.10μg / cm2 within 48h. During the transdermal process, the drug penetration is several to dozens of times that of ordinary patches, which can more effectively release drugs through the skin. Fig. 9 .

[0126] Embodiment 11

[0127] An improved dosage form of acupoint patch drug based on the theoretical system of traditional Chinese medicine, Evodia rutaecarpa soluble drug-loaded microneedles, and an in vitro transdermal intradermal retention and sustained drug release experiment.

[0128] In the transdermal experiment, the intradermal retention of Evodia rutaecarpa soluble microneedles acts as a drug reservoir to a certain extent. After the microneedle patch is peeled off, the intradermal retention amount is similar to a small drug reservoir, which can continue to release drugs after the microneedles are out of the body.

[0129] Based on Example 8, the permeation amount of the soluble microneedles of Evodia rutaecarpa retained in the skin under transdermal conditions was investigated. The permeation amount retained in the skin after microneedles were peeled off was compared with that of normal microneedles. The microneedles of the low-dose group and the peeling group were pressed on the ex vivo mouse skin for 5 minutes each, and then the mouse skin with microneedles inserted was fixed between the dosing pool and the receiving pool of the vertical transdermal instrument (the effective receiving area in the receiving pool was 0.68 cm 2 ). The peeling group peeled off the patch after 1 hour of permeation. Inject 4 mL of ethanol-PEG400-normal saline (4:3:3) into the receiving pool. Keep the temperature close to the skin temperature at 32°C and stir magnetically. Sampling time points are (2h, 4h, 6h, 8h, 10h, 12h, 18h, 24h, 48h). Take 1.5 mL of solution at each sampling point, pass through a 0.22μm filter membrane, wait for the liquid phase, and replenish the liquid. See Fig.10 .

[0130] The results show that during the transdermal administration of Evodia rutaecarpa soluble microneedles, after the microneedles are peeled off, the drugs remaining in the skin act as a drug reservoir to a certain extent in the subsequent period of time, and the drugs retained in the skin can still undergo a certain transdermal release.

Claims

1. A method for preparing soluble microneedles of Evodia rutaecarpa for acupoint administration, characterized in that: The effective biological components in Evodia rutaecarpa are modified to obtain a solid dispersion, which is then mixed with a needle tip matrix to prepare a drug-loaded microneedle; wherein the effective biological components in Evodia rutaecarpa are evodiamine (EVO) or rutaecarpine (RUT), and the structural formulas are 2. The method for preparing the soluble microneedle of Evodia rutaecarpa for acupoint administration according to claim 1, characterized in that: The solid dispersion is prepared by adding evodiamine (EVO) or rutaecarpine (RUT) into a solvent and stirring until completely dissolved. After dissolution, polyvinyl pyrrolidone (PVP K30) is added to the system under stirring conditions, and then rotary evaporation is performed to obtain a viscous product, which is then dried, ground, and sieved to obtain a solid dispersion.

3. The method for preparing the soluble microneedle of Evodia rutaecarpa for acupoint administration according to claim 2, characterized in that: The solid dispersion is prepared by adding evodiamine (EVO) or rutaecarpine (RUT) into a solvent, respectively, and magnetically stirring for 60 minutes in a 60°C water bath until the drug is completely dissolved in the solvent. PVP K30 is then added and magnetically stirred for 2 hours. The solution is rotary evaporated, and ethanol is recovered under reduced pressure until it becomes viscous. The solution is then transferred to a 60°C vacuum drying oven and dried for 12 hours. The solid is taken out, ground into powder using a mortar, and passed through a 100-mesh sieve to obtain a solid dispersion.

4. The method for preparing the soluble microneedles of Evodia rutaecarpa for acupoint administration according to claim 2 or 3, characterized in that: The evodiamine (EVO) and the solvent are mixed at a ratio of (1:5; w / v); the rutaecarpine (RUT) and the solvent are mixed at a ratio of (2:5; w / v); the evodiamine (EVO) and polyvinyl pyrrolidone (PVP K30) are mixed at a mass ratio of (1:1-8; w / w); and the rutaecarpine (RUT) and polyvinyl pyrrolidone PVP K30 are mixed at a mass ratio of (1:1-8; w / w).

5. The method for preparing the soluble microneedles of Evodia rutaecarpa for acupoint administration according to claim 2 or 3, characterized in that: The solvent for preparing the solid dispersion is anhydrous ethanol.

6. The method for preparing the soluble microneedles of Evodia rutaecarpa for acupoint administration according to claim 1, characterized in that: The solid dispersion, needle tip matrix and water are mixed to form a drug-loaded needle tip solution, wherein the solid dispersion and the needle tip solution are mixed in a mass to volume ratio (20-40:1; w / v); the needle tip matrix is ​​selected from a combination of two or more of polyvinyl pyrrolidone (PVP K30), enzymatically cleaved oligomeric sodium hyaluronate (HA), and chondroitin sulfate (CS).

7. The method for preparing the soluble microneedles of Evodia rutaecarpa for acupoint administration according to claim 1, characterized in that: The microneedles were prepared by centrifugation. The needle tip solution was added to the microneedle mold, centrifuged at 3000 rpm for 5 minutes, rotated 90 degrees for centrifugation, and repeated 4 times to make the mold rotate a full circle. The mold was placed at 25°C for drying for 30 minutes, and the backing matrix solution was added. After centrifugation for 5 minutes, the solution was rotated 180 degrees and centrifuged for another 5 minutes. The backing matrix was then added and placed in a vacuum drying oven. It was dried at 25°C for 24 hours and taken out after solidification.

8. The method for preparing the soluble microneedles of Evodia rutaecarpa for acupoint administration according to claim 7, characterized in that: The volume ratio of the needle tip solution to the backing matrix solution is 1:

5.

9. A soluble microneedle of Evodia rutaecarpa for acupoint administration prepared by the method of claim 1, characterized in that: The Evodia rutaecarpa soluble microneedles for acupoint administration prepared by the method of claim 1 have sharp needle bodies, good mechanical strength, and neatly arranged morphological arrays.

10. An application of the soluble microneedle of Evodia rutaecarpa for acupoint administration according to claim 9, characterized in that: The application of the acupoint-administered Evodia rutaecarpa soluble microneedles in the preparation of a patch medicine for treating oral ulcers.

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