A blood-activating and stasis-resolving, sinew-strengthening and bone-strengthening external-use plaster and a preparation process thereof

Abstract

The application provides a blood-activating and stasis-resolving external use plaster and a preparation process thereof, and the plaster is a microneedle-plaster composite preparation which is composed of a drug active ingredient, a sustained-release agent, a penetrating agent, an absorbent and a humectant. The drug active ingredient includes various Chinese herbal medicines such as Dysosma Versipellis, frankincense and myrrh, and the absorbent is a mixture of polyglutamic acid and snail polypeptide solution. The preparation process includes the steps of crushing medicinal materials, extracting with ethanol, concentrating and collecting cream, preparing a mixed solution, preparing a microneedle-plaster composite preparation and coating and the like. The snail polypeptide solution is obtained by enzymolysis of snail homogenate, and the humectant can be selected from hesperidin, glycerol and the like. The dissolvable material is selected from polylactic acid-hydroxyacetic acid copolymer and the like, and the ultrasonic power and frequency and the stirring speed of ultrasonic stirring all have certain ranges. The plaster has the effects of activating blood and resolving stasis and strengthening muscles and bones, and is suitable for external use. The preparation process is simple and feasible, and has high practical value and application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine preparation, in particular to a paste for external use with the functions of promoting blood circulation for removing blood stasis and strengthening muscles and bones and a preparation process thereof. BACKGROUND

[0002] In the field of traditional Chinese medicine, the paste for external use with the functions of promoting blood circulation for removing blood stasis and strengthening muscles and bones is one of the important means for treating common diseases such as traumatic injury and rheumatic bone disease. With the increasing attention to health and the deepening understanding of the advantages of traditional Chinese medicine, the market demand for such pastes for external use continues to grow. However, the related products on the current market have exposed many problems in actual application, which limits their further development and promotion.

[0003] In modern life, diseases such as traumatic injury and rheumatic bone pain are very common. These diseases not only bring physical pain to patients, but also can have a serious impact on their daily life and work. At present, for such diseases, the paste for external use with the functions of promoting blood circulation for removing blood stasis and strengthening muscles and bones is a common means of clinical treatment and daily health care.

[0004] Although traditional pastes for external use are widely used, they have many shortcomings. First, the transdermal absorption efficiency of drugs is low. Because the stratum corneum of the skin is a natural barrier, most of the drugs in traditional pastes are difficult to effectively penetrate this barrier and enter the deep tissues to play a role, resulting in low drug utilization rate. Many drug ingredients are wasted on the surface of the skin and cannot fully exert their therapeutic effect, which greatly reduces the therapeutic effect.

[0005] Second, the drug release rate is difficult to control. Most traditional pastes have a faster drug release rate at the initial stage of application. A large amount of drugs may reach the skin surface in a short time, but the drug release rate then rapidly decreases, making it difficult to maintain a stable blood drug concentration and continuously and effectively exert a therapeutic effect for a long time. This requires frequent replacement of the paste, which is inconvenient for patients and increases the cost of treatment.

[0006] Third, the drug ingredients are limited. The formula of traditional pastes is relatively fixed, and the types and quantities of drugs contained are limited, making it difficult to achieve a more comprehensive and ideal therapeutic effect by combining the synergistic effects of multiple drugs. For some complex diseases, a single or a small number of drug ingredients may not meet the treatment needs, affecting the targeting and effectiveness of treatment.

[0007] In order to overcome these problems, people have tried various improvement methods. For example, some chemical penetration enhancers are added to improve the transdermal absorption of drugs, but these chemicals may irritate the skin and cause allergic reactions and other adverse reactions. At the same time, some improvement methods are complex and costly in the operation process, which is not conducive to large-scale popularization and application.

[0008] Therefore, it has important practical significance to develop an external paste capable of improving the transdermal absorption efficiency of drugs, precisely controlling the drug release speed, comprehensively synergizing multiple drugs and being safe and reliable. SUMMARY

[0009] In view of this, the present application provides a blood-activating and stasis-removing external paste for strengthening muscles and bones and a preparation process thereof, which solves the above problems.

[0010] The technical scheme of the present application is as follows: an external paste for activating blood and removing stasis and strengthening muscles and bones, which is a microneedle-paste composite preparation coated on a backing material, wherein the microneedle-paste composite preparation is a paste medicine with microneedles, and the paste medicine is prepared from the following raw materials in percentage by weight: 40-60% of a drug active ingredient, 5-10% of a sustained-release agent, 2-4% of a penetration agent, 1-3% of an absorption agent, and the balance of a humectant; the drug active ingredient comprises the following raw materials in parts by weight: 10-14 parts of Dysosma Versipellis, 10-20 parts of Olibanum, 10-20 parts of Myrrh, 5-15 parts of Dragon's Blood, 15-30 parts of Drynaria, 15-30 parts of Eucommia Ulmoides, 10-20 parts of Radix Hedysari, 10-20 parts of Chuanxiong, 5-15 parts of Safflower, 8-16 parts of Radix Lamiophlomis, 10-20 parts of Sambucus, 6-12 parts of Radix Lamiophlomis, 10-18 parts of Rhizoma Pyrrosiae, and 3-8 parts of Borneol; and the absorption agent is a polyl-glutamic acid and snail polypeptide liquid in a mass ratio of (0.3-1.5):(6-10).

[0011] Further, the paste medicine is prepared from the following raw materials in percentage by weight: 40-60% of a drug active ingredient, 5-10% of a sustained-release agent, 2-4% of a penetration agent, 1-3% of an absorption agent, and the balance of a humectant.

[0012] Further, the drug active ingredient comprises the following raw materials in parts by weight: 12 parts of Dysosma Versipellis, 15 parts of Olibanum, 15 parts of Myrrh, 10 parts of Dragon's Blood, 22 parts of Drynaria, 28 parts of Eucommia Ulmoides, 15 parts of Radix Hedysari, 15 parts of Chuanxiong, 10 parts of Safflower, 12 parts of Radix Lamiophlomis, 15 parts of Sambucus, 9 parts of Radix Lamiophlomis, 15 parts of Rhizoma Pyrrosiae, and 5 parts of Borneol.

[0013] Further, the sustained-release agent is cyclodextrin, radix astragali gum and sophora japonica gum in a mass ratio of (12-25):(4-8):(5-10).

[0014] Further, the penetration agent is limonene, gingerol and eucalyptol in a mass ratio of (5-15):(4-8):(5-10).

[0015] Further, the snail polypeptide solution is that the snail is placed in a homogenizer, deionized water of 3-5 times of mass is added for homogenizing treatment, snail homogenate solution is obtained, complex protease is added, enzyme hydrolysis reaction is carried out at pH of 7.5-8.5 and temperature of 35-40 DEG C for 2-6 h, enzyme hydrolysis solution is obtained, heating is carried out to 80-90 DEG C, is maintained for 10-15 min, enzyme reaction is terminated, centrifugal separation is carried out, supernatant is collected, ultrafiltration and dialysis are carried out to obtain snail polypeptide solution with molecular weight <1 kD, and the mechanism is mainly to interfere with the lipid arrangement of cuticle or cause lipid extraction, thereby reducing the skin barrier function.

[0016] Further, the humectant is selected from any one or several combinations of hesperidin, glycerol, propylene glycol, sorbitol and vaseline.

[0017] Further, a preparation process of an external use plaster for activating blood and removing stasis and strengthening sinews and bones comprises the following steps:

[0018] S1, the medicinal materials of Gromwell, St. John's wort, earth Yuan, Stretches the sinew grass, Anemone altaica, Drynaria fortunei, Eucommia ulmoides, Radix Achyranthis Bidentatae, Chuanxiong, safflower are washed, dried and then crushed into coarse powder, which is sieved through a 60-80 mesh sieve for use; Frankincense, Myrrh, Dragon's Blood and Borneol are crushed into fine powder, which is sieved through a 100-120 mesh sieve for use;

[0019] S2, an ethanol solution with a mass concentration of 70-90% is added to the above medicinal materials, and the mixture is soaked for 8-12 h, heated to 50-80 DEG C and extracted for 3-5 times, each time for 2-4 h, and the extract is combined;

[0020] S3, the extract is concentrated to a relative density of 1.25-1.33 g / cm 3 at 60-80 DEG C under reduced pressure-0.05~-0.1 MPa, and the extract is collected to obtain an extract;

[0021] S4, the soluble material is dissolved in dichloromethane with a mass-volume ratio of 1:5-10, stirred uniformly to form a uniform solution, and then the extract of S3 is added, and ultrasonic stirring is carried out for 2-5 h to obtain a mixed solution, the mixed solution is slowly injected into a microneedle mold, the content of the extract in the microneedle accounts for 10-30% of the total mass of the microneedle, centrifugation is carried out at a speed of 3000-5000 rpm for 5-10 min, and then vacuum drying is carried out at a vacuum degree of-0.08~-0.1 MPa and a temperature of 30-40 DEG C, demolding is carried out, and a microneedle-plaster composite preparation is prepared;

[0022] S5, the microneedle-plaster composite preparation is coated on the backing material to obtain an external use plaster.

[0023] Further, the soluble material of S4 is selected from polylactic acid-glycolic acid copolymer (PLGA), hyaluronic acid or chitosan.

[0024] Further, the ultrasonic stirring ultrasonic power is 500-800W, the ultrasonic frequency is 20-50kHz, and the stirring rate is 500-800rpm.

[0025] Compared with the prior art, the present application has the following advantages:

[0026] The active pharmaceutical ingredients of the external use plaster of the present application are selected from various precious medicinal materials such as Dysosma Versipellis and Frankincense, which play the effects of activating blood and resolving stasis, strengthening muscles and bones in cooperation with each other. The proportion of each component in the plaster medicine is carefully adjusted, and the proportion of the active pharmaceutical ingredients, the sustained-release agent, the penetrant, the absorbent and the humectant is scientific and reasonable. The micro-needle-plaster composite preparation is adopted to increase the drug permeability by penetrating the stratum corneum of the skin, and to overcome the problem of difficult absorption of traditional plasters. The sustained-release agent composed of specific proportions of cyclodextrin, radix astragali and sophora japonica gum precisely controls the drug release rate and maintains long-term effective blood drug concentration. The penetrant composed of limonene, gingerol and eucalyptol and the absorbent composed of polyglutamic acid and snail polypeptide liquid synergistically act to improve the drug absorption and utilization rate. In the preparation process, the optimized extraction method of medicinal materials fully retains the efficacy, the soluble material is selected for the preparation of micro-needles, and the ultrasonic stirring, centrifugation and other processes are adopted to ensure the quality and performance of the micro-needles and precisely control the extract content. In addition, the plaster mainly contains natural medicinal materials, has high safety, reduces adverse reactions, the selection of various humectants provides flexibility for the formula design, meets different needs, and has a wide range of applications. DETAILED DESCRIPTION

[0027] In order to better understand the technical content of the present application, the following specific examples are provided to further illustrate the present application.

[0028] The experimental methods used in the embodiments of the present application are conventional methods unless otherwise specified.

[0029] The materials, reagents and the like used in the embodiments of the present application can be obtained from commercial channels unless otherwise specified.

[0030] The basic conditions of the main raw materials are introduced according to Chinese Pharmacopoeia, Chinese Herbal Medicine Dictionary, Chinese Herbal Medicine and the like.

[0031] Glehnia Root: Nature and flavor meridian: warm in nature, pungent in taste. Belongs to kidney meridian and liver meridian. Function and treatment: dispelling wind and dampness, relaxing muscles and promoting blood circulation, removing blood stasis and swelling, detoxifying and relieving pain.

[0032] Sambucus: Nature and flavor meridian: sweet and bitter in taste, neutral in nature. Belongs to liver meridian. Function and treatment: bone setting and tendon continuation, promoting blood circulation and relieving pain, dispelling wind and dampness.

[0033] Earth Yuan: Nature and flavor meridian: salty in taste, cold in nature; slightly toxic. Belongs to liver meridian. Function and treatment: breaking blood stasis, continuing bones and tendons.

[0034] Rhizoma Drynariae: Pungent, bitter, warm. Liver, spleen, kidney meridians. Expelling wind and dampness, relaxing the sinews. Used for joint pain, flexion and extension of the limbs.

[0035] Rhizoma Drynariae: Pungent, bitter, warm. Liver, spleen, kidney meridians. Expelling wind and dampness, relaxing the sinews. Used for joint pain, flexion and extension of the limbs.

[0036] Rhizoma Drynariae: Pungent, bitter, warm. Liver, spleen, kidney meridians. Expelling wind and dampness, relaxing the sinews. Used for joint pain, flexion and extension of the limbs.

[0037] Rhizoma Drynariae: Pungent, bitter, warm. Liver, spleen, kidney meridians. Expelling wind and dampness, relaxing the sinews. Used for joint pain, flexion and extension of the limbs.

[0038] Rhizoma Drynariae: Pungent, bitter, warm. Liver, spleen, kidney meridians. Expelling wind and dampness, relaxing the sinews. Used for joint pain, flexion and extension of the limbs.

[0039] Rhizoma Drynariae: Pungent, bitter, warm. Liver, spleen, kidney meridians. Expelling wind and dampness, relaxing the sinews. Used for joint pain, flexion and extension of the limbs.

[0040] Rhizoma Drynariae: Pungent, bitter, warm. Liver, spleen, kidney meridians. Expelling wind and dampness, relaxing the sinews. Used for joint pain, flexion and extension of the limbs.

[0041] Rhizoma Drynariae: Pungent, bitter, warm. Liver, spleen, kidney meridians. Expelling wind and dampness, relaxing the sinews. Used for joint pain, flexion and extension of the limbs.

[0042] Rhizoma Drynariae: Pungent, bitter, warm. Liver, spleen, kidney meridians. Expelling wind and dampness, relaxing the sinews. Used for joint pain, flexion and extension of the limbs.

[0043] Rhizoma Drynariae: Pungent, bitter, warm. Liver, spleen, kidney meridians. Expelling wind and dampness, relaxing the sinews. Used for joint pain, flexion and extension of the limbs.

[0044] Rhizoma Drynariae: Pungent, bitter, warm. Liver, spleen, kidney meridians. Expelling wind and dampness, relaxing the sinews. Used for joint pain, flexion and extension of the limbs. Example 1

[0045] A kind of external use paste of blood-activating and stasis-removing, strong sinew and bone, the external use paste is that micro-needle-paste compound preparation is coated on backing material, the micro-needle-paste compound preparation is paste medicine with micro-needle, the paste medicine includes the following weight percentage raw materials and is prepared: medicinal active ingredient 40%, slow-release agent 5%, penetrant 2%, absorbent 1%, and the balance is humectant.

[0046] The pharmaceutically active ingredients include the following raw materials by weight: Typhonium giganteum 10 parts, Frankincense 10 parts, Myrrh 10 parts, Dragon's Blood 5 parts, Drynaria 15 parts, Eucommia ulmoides 15 parts, Radix Achyranthis Bidentatae 10 parts, Chuanxiong 10 parts, Safflower 5 parts, Radix Lamiophlomis rotata 8 parts, Sambucus williamsii 10 parts, Radix Rubi 6 parts, Rhizoma Pyrrosiae 10 parts, Borneol 3 parts.

[0047] The sustained-release agent is cyclodextrin, tragacanth gum and locust bean gum at a mass ratio of 12:4:5.

[0048] The penetrating agent is limonene, gingerol and eucalyptol at a mass ratio of 5:4:5.

[0049] The absorption agent is polyglutamic acid and snail polypeptide liquid at a mass ratio of 0.3:6.

[0050] The snail polypeptide liquid is obtained by homogenizing snails in a homogenizer with 3 times the mass of deionized water, adding complex protease, performing enzymatic reaction at pH 7.5 and 35°C for 2 hours, heating to 80°C for 10 minutes to terminate the enzyme reaction, centrifugal separation, and collecting the supernatant, ultrafiltration and dialysis to obtain a snail polypeptide liquid with a molecular weight of <1 kD.

[0051] The humectant is selected from hesperidin. Example 2

[0052] An external use plaster for promoting blood circulation and removing blood stasis and strengthening tendons and bones, the external use plaster is a microneedle-plaster composite preparation coated on a backing material, the microneedle-plaster composite preparation is a plaster medicine with microneedles, and the plaster medicine is prepared from the following raw materials by weight percentage: pharmaceutically active ingredients 60%, sustained-release agent 10%, penetrating agent 4%, absorption agent 3%, and the balance is a humectant.

[0053] The pharmaceutically active ingredients include the following raw materials by weight: Typhonium giganteum 14 parts, Frankincense 20 parts, Myrrh 20 parts, Dragon's Blood 15 parts, Drynaria 30 parts, Eucommia ulmoides 30 parts, Radix Achyranthis Bidentatae 20 parts, Chuanxiong 20 parts, Safflower 15 parts, Radix Lamiophlomis rotata 16 parts, Sambucus williamsii 20 parts, Radix Rubi 12 parts, Rhizoma Pyrrosiae 18 parts, Borneol 8 parts.

[0054] The sustained-release agent is cyclodextrin, tragacanth gum and locust bean gum at a mass ratio of 25:8:10.

[0055] The penetrating agent is limonene, gingerol and eucalyptol at a mass ratio of 15:8:10.

[0056] The absorption agent is polyglutamic acid and snail polypeptide liquid at a mass ratio of 1.5:10.

[0057] The snail polypeptide solution is obtained by placing snails in a homogenizer, adding 5 times the mass of deionized water to homogenize, obtaining a snail homogenate, adding complex protease, performing enzymatic reaction at pH 8.5 and 40℃ for 6h, obtaining an enzymatic hydrolysate, heating to 90℃ for 15min to terminate the enzyme reaction, centrifugal separation, collecting the supernatant, ultrafiltration and dialysis to obtain a snail polypeptide solution with a molecular weight of <1kD.

[0058] The humectant is selected from glycerol. Example 3

[0059] A blood-activating and stasis-removing external use plaster for strengthening muscles and bones, the external use plaster is a microneedle-plaster composite preparation coated on a backing material, the microneedle-plaster composite preparation is a plaster medicine with microneedles, and the plaster medicine is prepared from the following raw materials in percentage by weight: 50% of a pharmaceutical active ingredient, 8% of a sustained-release agent, 3% of a penetration agent, 2% of an absorption agent, and the balance of a humectant.

[0060] The pharmaceutical active ingredient includes the following raw materials in parts by weight: 12 parts of Dysosma Versipellis, 15 parts of Olibanum, 15 parts of Myrrh, 10 parts of Dragon's Blood, 22 parts of Drynaria, 28 parts of Eucommia Ulmoides, 15 parts of Radix Cyathulae, 15 parts of Chuanxiong Rhizome, 10 parts of Safflower, 12 parts of Radix Trifolii, 15 parts of Sambucus, 9 parts of Radix Rubi, 15 parts of Radix Pyrrosiae, and 5 parts of Borneol.

[0061] The sustained-release agent is cyclodextrin, radix astragali gum and sophora japonica gum at a mass ratio of 20:6:8.

[0062] The penetration agent is limonene, gingerol and eucalyptol at a mass ratio of 10:6:8.

[0063] The absorption agent is polyglutamic acid and snail polypeptide solution at a mass ratio of 0.9:8.

[0064] The snail polypeptide solution is obtained by placing snails in a homogenizer, adding 5 times the mass of deionized water to homogenize, obtaining a snail homogenate, adding complex protease, performing enzymatic reaction at pH 8.5 and 40℃ for 6h, obtaining an enzymatic hydrolysate, heating to 90℃ for 15min to terminate the enzyme reaction, centrifugal separation, collecting the supernatant, ultrafiltration and dialysis to obtain a snail polypeptide solution with a molecular weight of <1kD.

[0065] The humectant is selected from glycerol.

[0066] The above examples 1-3 are prepared by the following preparation process:

[0067] S1, the Radix Trifolii, Sambucus, Radix Rubi, Radix Pyrrosiae, Dysosma Versipellis, Drynaria, Eucommia Ulmoides, Radix Cyathulae, Chuanxiong Rhizome, Safflower medicinal materials are washed, dried and crushed into coarse powder, and then sieved through a 70-mesh sieve for use; Olibanum, Myrrh, Dragon's Blood and Borneol are crushed into fine powder and sieved through a 100-mesh sieve for use;

[0068] S2, add an ethanol solution with a mass concentration of 80% to the above medicinal material, soak for 8-12 h, and extract 4 times at 70°C, each time for 4 h, and combine the extract;

[0069] S3, concentrate the extract at 70°C and under reduced pressure of-0.08 MPa to a relative density of 1.30 g / cm 3 , collect the extract, and obtain an extract;

[0070] S4, dissolve polylactic acid-glycolic acid copolymer in dichloromethane at a mass-volume g / mL ratio of 1:8, stir uniformly to form a uniform solution, then add the extract of S3, ultrasonically stir for 3 h at an ultrasonic power of 700 W, an ultrasonic frequency of 30 kHz, and a stirring rate of 600 rpm, obtain a mixed solution, slowly inject the mixed solution into a microneedle mold, the extract content accounts for 20% of the total mass of the microneedle, centrifuge at a speed of 4000 rpm for 5-10 min, then vacuum dry at a vacuum degree of-0.1 MPa and a temperature of 35°C, demold, and obtain a microneedle-paste composite preparation;

[0071] S5, coat the above microneedle-paste composite preparation on a backing material to obtain an external-use paste.

[0072] Comparative Example 1

[0073] The difference between this comparative example and Example 3 is that the external-use paste is directly coated on the backing material.

[0074] Comparative Example 2

[0075] The difference between this comparative example and Example 3 is that the active ingredient of the paste does not contain an absorbent.

[0076] Comparative Example 3

[0077] The difference between this comparative example and Example 3 is that the active ingredient of the paste does not contain a sustained-release agent.

[0078] Comparative Example 4

[0079] The difference between this comparative example and Example 3 is that the active ingredient of the paste does not contain a permeation agent.

[0080] Test Example 1-In Vitro Transdermal Experiment

[0081] 1. Skin preparation: remove the subcutaneous fat of fresh isolated pig skin, rinse with physiological saline, and then absorb the surface moisture with filter paper. Fix the treated pig skin between the supply pool and the receiving pool of the Franz diffusion cell to ensure that the skin is tightly attached and there is no leakage.

[0082] 2. Patch placement: The test patches of Examples 1-3 and Comparative Examples 1-4 were accurately cut to 2.5 cm and uniformly attached to the surface of the skin, so as to be in full contact with the skin.

[0083] 3. Replacement of receiving solution and sampling: 5 mL of receiving solution was added to the receiving pool, which was kept at a temperature of 37°C ± 0.5°C and stirred at a constant speed (500 rpm). At 0.5, 1, 2, 4, 6, 8, 12 and 24 hours, 0.5 mL of receiving solution was taken from the receiving pool, while an equal amount of fresh receiving solution was added to maintain the constant volume of the receiving solution.

[0084] 4. Drug content determination: The removed receiving solution was centrifuged and filtered, and the drug content therein was determined by high performance liquid chromatography. The cumulative transdermal amount of the drug at different time points was calculated according to the standard curve, and the transdermal rate of the drug was calculated.

[0085] 5. Test results

[0086]

[0087] Test Example 2 - Pharmacodynamic experiment

[0088] 1. Experimental animals

[0089] Healthy adult rats weighing 180-220 g (rats) were selected, with half male and half female.

[0090] 2. Establishment of disease model

[0091] Fracture model: A rat fracture model was used. Under anesthesia, a fracture was made in the middle of the femur of the animal using surgical instruments, and then fixed with a splint to simulate a fracture injury.

[0092] Arthritis model: A collagen-induced arthritis (CIA) model was used. An emulsion of bovine type II collagen and complete Freund's adjuvant was injected into the footpads of the animals to induce arthritis. After modeling, the animal's joint swelling, pain and other symptoms were observed to confirm the success of the model.

[0093] 3. Grouping and administration

[0094] The animals were randomly divided into an experimental group (patch administration group), a positive control group (commercial Yunnan Baiyao ointment) and a negative control group (blank patch group), with no less than 10 animals in each group.

[0095] Experimental group: The test patches of Examples 1-3 and Comparative Examples 1-4 were applied to the fracture site of the animals according to an area of 3*3 cm 2 and a dose of 400 mg, and were replaced once a day.

[0096] Positive control group: The same method and dosage of commercially available Yunnan Baiyao plaster was given.

[0097] Negative control group: The same method and dosage of commercially available Yunnan Baiyao plaster was given.

[0098] 4. Observation index

[0099] (1) Observation index of fracture model

[0100] Imaging examination: On the 7th, 14th, and 21st days after administration, X-ray examination was performed on the animals to observe the healing of the fracture, measure the area of callus formation, and the degree of fracture line healing.

[0101] Biomechanical test: After 28 days of administration, biomechanical test was performed on the fractured limbs of the animals to determine the maximum load, elastic modulus, and other parameters of the bone, and to evaluate the recovery of the mechanical properties of the bone.

[0102] (2) Observation index of arthritis model

[0103] Joint swelling degree: The circumference of the animal's joint was measured using a vernier caliper, 2-3 times per week, and the joint swelling degree was calculated to evaluate the degree of joint inflammation.

[0104] Pain behavior test: The hot plate method or pressure pain method was used to regularly determine the pain threshold of the animals, observe the changes in the animals' pain behavior, and evaluate the analgesic effect of the plaster.

[0105] Histopathological examination: After the experiment, the joint tissue of the animals was taken for pathological section, and the pathological changes of the articular cartilage, synovial membrane, and other tissues were observed, and the improvement of arthritis was evaluated by histological scoring.

[0106] 4. Result data

[0107] Fracture model:

[0108]

[0109] On the 14th and 21st days, the callus formation area of Example 3 was the largest, followed by Example 2 and Example 1, which were significantly larger than the comparative example group, the positive control group, and the negative control group. This indicates that the plaster of the examples can more effectively promote the early formation of callus, with Example 3 being the best.

[0110] On the 28th day, the fracture line of Example 3 completely disappeared, reaching basic healing, and Examples 1 and 2 also had obvious healing, with the fracture line nearly disappearing or almost invisible. The fracture line of the comparative example group was blurred, and the healing was better, but still not as good as the example group. The fracture line of the positive control group was still visible, and the healing degree was general, and the fracture line of the negative control group was clear, and the healing was slow.

[0111] The bone maximum load of Example 3 was the highest, 42.0±3.0N, indicating that it had the best effect on the recovery of bone strength after fracture. Examples 1 and 2 also had higher load values, followed by the comparative group, and the positive and negative control groups were lower.

[0112] Arthritis model:

[0113]

[0114] At weeks 2 and 4, the joint swelling of Example 3 decreased most significantly, to only 5.0±1.0% at week 4, and Examples 1 and 2 also had good detumescence effects. The joint swelling of the comparative group was higher than that of the example groups, but lower than that of the positive and negative control groups.

[0115] The pain threshold of Example 3 was the highest, 25.0±2.5s, indicating that it could most effectively relieve arthritis pain. Examples 1 and 2 also had higher pain thresholds, followed by the comparative group, and the positive and negative control groups were lower.

[0116] The histological score of Example 3 was the lowest, 1.5±0.2, indicating that the degree of inflammation and damage to the joint tissue was the lightest. Examples 1 and 2 also had lower scores, and the score of the comparative group was higher than that of the example groups but lower than that of the positive and negative control groups.

[0117] Test Example 3 - Clinical Test

[0118] 1. Test Design

[0119] A randomized, double-blind, parallel control clinical test design was used.

[0120] 2. Selection of Subjects

[0121] Patients meeting the following criteria were recruited

[0122] (1) Aged 18-65 years old, male or female.

[0123] (2) Suffering from diseases such as fractures, sprains, osteoarthritis, rheumatoid arthritis, etc. that meet the treatment categories of promoting blood circulation and removing blood stasis and strengthening tendons and bones, and the disease condition is in the stable period.

[0124] (3) Patients voluntarily participate in the test and sign the informed consent form.

[0125] Exclusion Criteria

[0126] (1) Allergic to any ingredient in the plaster.

[0127] (2) Combined with serious diseases of important organs such as heart, liver, kidney, etc.

[0128] (3) Those who are currently using other drugs or treatments that may affect the efficacy evaluation of this patch.

[0129] 3. Grouping and Dosing

[0130] The subjects were randomly divided into experimental group and control group, with no less than 50 subjects in each group.

[0131] (1) Experimental group: The test patch was given and applied according to the prescribed application site and method. The patch was changed once a day and used continuously for 4 weeks as a course of treatment.

[0132] (2) Control group: A placebo patch that looks the same as the test patch but does not contain any drug ingredients was given. The method of use and course of treatment were the same as those of the experimental group.

[0133] 4. Observation indicators

[0134] (1) Clinical symptom scoring: Before treatment, and at the second and fourth weeks after treatment, the patients' clinical symptoms such as pain, swelling, and limited joint movement were scored. The visual analogue scale (VAS) was used to assess the degree of pain, and the swelling and limited joint movement were assessed according to relevant standards.

[0135] (2) Imaging examination: Before treatment and 4 weeks after treatment, patients will undergo X-ray, CT or MRI examinations to observe fracture healing, changes in joint structure, etc., and will be evaluated by professional imaging physicians.

[0136] (3) Safety indicators: During the entire trial, the patient's adverse reactions, including skin allergies, local irritation and other symptoms, were closely observed. At the same time, the patient's vital signs (body temperature, blood pressure, heart rate, etc.) and laboratory test indicators (such as blood routine, liver and kidney function, etc.) were recorded to evaluate the safety of the patch.

[0137] 5. Clinical trial results

[0138]

[0139] This clinical trial analyzed indicators such as pain VAS scores, improvement rates of joint motion limitation, and radiographic improvement rates and found that the pain VAS scores of the experimental, comparative, positive, and negative control groups were similar before treatment. After treatment, pain VAS scores decreased in all groups. The Example group showed outstanding results in terms of pain relief, improvement of joint motion limitation, and radiographic improvement. Example 3 showed the best results, with a pain score of only 1.5±0.3 after 4 weeks of treatment, an improvement rate of 85.6%±8.3% in joint motion limitation, and an improvement rate of 78.6%±7.5% in radiographic improvement.

[0140] The above merely provides the preferred embodiment of the present application, and is not used to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.

Claims

1. An external-use plaster for promoting blood circulation, removing blood stasis, strengthening tendons and bones, characterized by: The topical patch is prepared by coating a microneedle-patch composite preparation on a backing material. The microneedle-patch composite preparation is a patch drug with microneedles. The patch drug is prepared from the following raw materials in weight percentage: 40-60% active pharmaceutical ingredient, 5-10% sustained-release agent, 2-4% penetrant, 1-3% absorbent, and the balance is a moisturizer; the active pharmaceutical ingredient is prepared from the following raw materials in weight parts: 10-14 parts of ophiopogon japonicus, 10 parts of frankincense, 10 parts of hyacinthium, 10 parts of iris root ... -20 parts, myrrh 10-20 parts, blood 5-15 parts, drynaria 15-30 parts, eucommia ulmoides 15-30 parts, Achyranthes bidentata 10-20 parts, Chuanxiong 10-20 parts, safflower 5-15 parts, 8-16 parts of safflower, elderberry 10-20 parts, earthworm 6-12 parts, 10-18 parts of smilax glabra, 3-8 parts of borneol, the absorbent is a polyglutamic acid and snail polypeptide solution in a mass ratio of (0.3-1.5): (6-10); The sustained-release agent is cyclodextrin, tragacanth gum and locust bean gum in a mass ratio of (12-25):(4-8):(5-10); The penetrants are limonene, gingerol and eucalyptol in a mass ratio of (5-15):(4-8):(5-10).

2. The external-use plaster for promoting blood circulation, removing blood stasis, strengthening tendons and bones according to claim 1, characterized in that: The patch medicine is prepared from the following raw materials in percentage by weight: 40-60% of active pharmaceutical ingredient, 5-10% of sustained-release agent, 2-4% of penetrant, 1-3% of absorbent, and the balance of moisturizer.

3. The external-use plaster for promoting blood circulation, removing blood stasis, strengthening tendons and bones according to claim 1, characterized in that: The active ingredient of the medicine is prepared from the following raw materials in parts by weight: 12 parts of ophiopogon japonicus, 15 parts of frankincense, 15 parts of myrrh, 10 parts of dragon's blood, 22 parts of drynaria rhizome, 28 parts of eucommia bark, 15 parts of cyperus rotundus, 15 parts of ligusticum chuanxiong, 10 parts of safflower, 12 parts of radix polygoni lobatae, 15 parts of elderberry, 9 parts of earthworm, 15 parts of herba striatae, and 5 parts of borneol.

4. The external-use plaster for promoting blood circulation, removing blood stasis, strengthening tendons and bones according to claim 1, characterized in that: The snail polypeptide liquid is prepared by placing the snail in a homogenizer, adding 3-5 times the mass of deionized water for homogenization to obtain a snail homogenate, adding a composite protease, performing an enzymatic hydrolysis reaction at a pH of 7.5-8.5 and a temperature of 35-40°C for 2-6 hours to obtain an enzymatic hydrolyzate, heating the solution to 80-90°C and maintaining the reaction for 10-15 minutes, terminating the enzyme reaction, centrifuging, collecting the supernatant, ultrafiltration, and dialyzing to obtain a snail polypeptide liquid with a molecular weight of less than 1kD.

5. The external-use plaster for promoting blood circulation, removing blood stasis, strengthening tendons and bones according to claim 1, characterized in that: The moisturizing agent is selected from any one or a combination of hesperidin, glycerin, propylene glycol, sorbitol, and vaseline.

6. The preparation process of the external-use plaster for promoting blood circulation, removing blood stasis, strengthening tendons and strengthening bones as claimed in claim 1, characterized in that: The following steps are involved: S1. Wash and dry the herbs of Radix Angelicae Dahuricae, Radix Elderberry, Radix Echinops, Herba Lycopodii, Rhizoma Drynariae, Rhizoma Eucommiae, Rhizoma Cyathulae, Rhizoma Chuanxiong, and Flos Carthami, grind them into coarse powder, and pass through a 60-80 mesh sieve for later use; grind frankincense, myrrh, and dragon's blood into fine powder, and pass through a 100-120 mesh sieve for later use; S2. Add 70-90% ethanol solution to the above medicinal materials, let them soak for 8-12 hours, heat to 50-80°C and extract 3-5 times, each time for 2-4 hours, and combine the extracts; S3. Concentrate the extract at 60-80°C and reduced pressure of -0.05~-0.1MPa to a relative density of 1.25-1.33g / cm 3 , collect the paste and obtain the extract; S4, dissolving the soluble material in dichloromethane at a mass-to-volume ratio of g / mL of 1:5-10, stirring evenly to form a uniform solution, then adding the extract of S3, and ultrasonically stirring for 2-5 hours to obtain a mixed solution, slowly injecting the mixed solution into the microneedle mold, wherein the extract content accounts for 10-30% of the total mass of the microneedles, centrifuging at a rate of 3000-5000 rpm for 5-10 minutes, and then vacuum drying at a vacuum degree of -0.08 to -0.1 MPa and a temperature of 30-40°C, and demolding to obtain a microneedle-patch composite preparation; the soluble material is selected from polylactic acid-glycolic acid copolymer, hyaluronic acid, or chitosan; S5. Apply the above-mentioned microneedle-patch composite preparation on a backing material to obtain an external-use patch.

7. The preparation process of the external-use plaster for promoting blood circulation, removing blood stasis, strengthening tendons and strengthening bones as claimed in claim 6, characterized in that: The ultrasonic stirring has an ultrasonic power of 500-800 W, an ultrasonic frequency of 20-50 kHz, and a stirring rate of 500-800 rpm.

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