External plaster for promoting blood circulation to remove blood stasis and strengthening tendons and bones and preparation process thereof

Abstract

The invention provides an external plaster for promoting blood circulation to remove blood stasis and strengthening tendons and bones and a preparation process thereof, and the plaster adopts a microneedle-plaster composite preparation and is composed of active pharmaceutical ingredients, a sustained release agent, a penetrant, an absorbent and a humectant. The medicine active ingredients comprise multiple Chinese herbal medicines such as dysosma versipellis, frankincense and myrrh, and the absorbent is a mixture of polyglutamic acid and snail polypeptide liquid. The preparation process comprises the steps of medicinal material crushing, ethanol extraction, concentration and paste collection, preparation of a mixed solution, preparation of a microneedle-plaster composite preparation, coating and the like. Wherein the snail polypeptide liquid is obtained by enzymolysis of snail homogenate, and the humectant can be selected from hesperidin, glycerol and the like. The dissoluble material is selected from polylactic acid-glycolic acid copolymer and the like, and the ultrasonic power, the ultrasonic frequency and the stirring speed of ultrasonic stirring are within a certain range. The plaster has the effects of promoting blood circulation to remove blood stasis and strengthening tendons 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 invention relates to the technical field of drug preparation, and in particular to an external plaster for promoting blood circulation, removing blood stasis, strengthening tendons and bones, and a preparation process thereof. Background Art

[0002] In the field of traditional Chinese medicine, topical plasters that promote blood circulation, remove blood stasis, strengthen tendons and strengthen bones are one of the important means of treating common diseases such as traumatic injuries and rheumatic bone diseases. With the increase of people's attention to health and the in-depth understanding of the advantages of traditional Chinese medicine, the market demand for such topical plasters continues to grow. However, the relevant products on the market currently have exposed many problems in actual application, which limits their further development and promotion.

[0003] In modern life, injuries from falls, rheumatism and bone pain are extremely common. These diseases not only cause physical pain to patients, but also may have a serious impact on their daily life and work. At present, for such diseases, external plasters that promote blood circulation and remove blood stasis, strengthen tendons and bones are commonly used in clinical treatment and daily health care.

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

[0005] Second, the drug release rate is difficult to control. Most traditional patches release drugs quickly in the initial stage of application, and more drugs may reach the skin surface in a short period of time, but then the drug release rate drops rapidly, making it difficult to maintain a stable blood drug concentration and unable to continue to effectively exert a therapeutic effect over a long period of time. This requires frequent replacement of patches, which brings inconvenience to patients and also increases treatment costs.

[0006] Third, the drug ingredients are limited. The formula of traditional plasters is relatively fixed, and the types and quantities of drugs contained are limited. It is difficult to integrate the synergistic effects of multiple drugs to achieve a more comprehensive and ideal therapeutic effect. For some complex diseases, a single or a few drug ingredients may not meet the treatment needs, affecting the pertinence and effectiveness of the treatment.

[0007] In order to overcome these problems, people have tried a variety of improvement methods. For example, adding some chemical penetration enhancers to improve the transdermal absorption of drugs, but these chemicals may irritate the skin and cause adverse reactions such as allergies. At the same time, some improvement methods are more complicated in operation and have higher costs, which is not conducive to large-scale promotion and application.

[0008] Therefore, it is of great practical significance to develop a safe and reliable external patch that can improve the efficiency of drug transdermal absorption, accurately control the drug release rate, and integrate the synergistic effects of multiple drugs. Summary of the invention

[0009] In view of this, the present invention provides an external plaster for promoting blood circulation, removing blood stasis, strengthening tendons and bones and a preparation process thereof to solve the above problems.

[0010] The technical solution of the present invention is achieved as follows: an external patch for promoting blood circulation, removing blood stasis, strengthening tendons and strengthening bones, wherein the external patch is a microneedle-patch composite preparation coated on a backing material, wherein the microneedle-patch composite preparation is a patch drug with microneedles, and the patch drug comprises the following raw materials in weight percentage: 40-60% of active pharmaceutical ingredients, 5-10% of sustained-release agent, 2-4% of penetrant, 1-3% of absorbent, and the remainder is a moisturizer; the active pharmaceutical ingredients comprise the following raw materials in weight percentage: Materials: 10-14 parts of Illustris odoratus, 10-20 parts of frankincense, 10-20 parts of myrrh, 5-15 parts of Sanguisorba officinalis, 15-30 parts of Drynaria fortunei, 15-30 parts of Eucommia ulmoides, 10-20 parts of Achyranthes bidentata, 10-20 parts of Chuanxiong, 5-15 parts of Carthamus tinctorius, 8-16 parts of Herba schizonepetae, 10-20 parts of Elderberry, 6-12 parts of Earthworm, 10-18 parts of Herba Lycopodiellae, and 3-8 parts of Borneolum. The absorbent is a polyglutamic acid and snail polypeptide solution in a mass ratio of (0.3-1.5):(6-10).

[0011] Furthermore, the patch medicine is prepared from the following raw materials in weight percentage: 40-60% of active pharmaceutical ingredient, 5-10% of sustained-release agent, 2-4% of permeation agent, 1-3% of absorbent, and the balance is moisturizer.

[0012] Furthermore, the active ingredients of the medicine include the following raw materials in parts by weight: 12 parts of Illustris odoratus, 15 parts of frankincense, 15 parts of myrrh, 10 parts of Sanguisorba officinalis, 22 parts of Drynaria fortunei, 28 parts of Eucommia ulmoides, 15 parts of Achyranthes bidentata, 15 parts of Chuanxiong, 10 parts of Carthamus tinctorius, 12 parts of Herba schizonepetae schizonepetae, 15 parts of Elderberry, 9 parts of Earthworm, 15 parts of Herba Lycopodii, and 5 parts of Borneol.

[0013] Furthermore, the sustained-release agent is cyclodextrin, tragacanth gum and locust bean gum in a mass ratio of (12-25):(4-8):(5-10).

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

[0015] Furthermore, 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, and 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 solution, heating the solution to 80-90°C and maintaining the reaction for 10-15 minutes to terminate the enzyme reaction, centrifuging and separating, collecting the supernatant, ultrafiltrating and dialyzing to obtain a snail polypeptide liquid with a molecular weight of less than 1kD. The mechanism is mainly to interfere with the lipid arrangement of the stratum corneum or to cause lipid extraction, thereby reducing the skin barrier function.

[0016] Furthermore, the moisturizing agent is selected from any one or a combination of hesperidin, glycerin, propylene glycol, sorbitol, and vaseline.

[0017] Furthermore, a preparation process of an external plaster for promoting blood circulation, removing blood stasis, strengthening tendons and strengthening bones comprises the following steps:

[0018] S1. Wash and dry the Radix Angelicae Pubescentis, Radix Elderberry, Radix Glehniae, Herba Lycopodii, Rhizoma Drynariae, Eucommia Bark, Rhizoma Cyathulae, Rhizoma Chuanxiong, and Flos Carthami, grind them into coarse powder, and sieve through 60-80 mesh for later use; grind frankincense, myrrh, Sanguisorba Officinalis, and borneol into fine powder, and sieve through 100-120 mesh for later use;

[0019] S2. Add 70-90% ethanol solution to the above medicinal materials, soak for 8-12 hours, heat to 50-80°C, extract 3-5 times, each time for 2-4 hours, and combine the extracts;

[0020] S3, concentrate the extract at 60-80℃ and reduced pressure -0.05~-0.1MPa to a relative density of 1.25-1.33g / cm 3 , collect the paste and obtain the extract;

[0021] S4, dissolving the soluble material in dichloromethane with a mass volume g / mL ratio of 1:5-10, stirring evenly to form a uniform solution, then adding the extract of S3, and then ultrasonically stirring for 2-5h to obtain a mixed solution, slowly injecting the mixed solution into the microneedle mold, the extract content accounts for 10-30% of the total mass of the microneedle, centrifuging at a rate of 3000-5000rpm for 5-10min, and then vacuum drying at a vacuum degree of -0.08~-0.1MPa and a temperature of 30-40°C, demolding, and obtaining a microneedle-patch composite preparation;

[0022] S5. Apply the above-mentioned microneedle-patch composite preparation on a backing material to obtain an external patch.

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

[0024] Furthermore, 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.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The active pharmaceutical ingredients of the topical plaster of the present invention are selected from a variety of precious medicinal materials such as ophiopogon erythrorhizon and frankincense, which are mutually compatible and synergistic to play the effects of promoting blood circulation and removing blood stasis, strengthening tendons and bones, and the proportions of the various ingredients in the plaster medicine are carefully formulated, and the proportions of the active pharmaceutical ingredients, sustained-release agents, penetrants, absorbents and moisturizers are scientific and reasonable. The microneedle-patch composite preparation is adopted, and the microneedles penetrate the stratum corneum of the skin to increase the permeability of the drug, overcoming the problem of difficult absorption of traditional plasters; the sustained-release agent composed of a specific proportion of cyclodextrin, tragacanth gum and locust bean gum accurately controls the drug release rate and maintains a 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 work synergistically to improve the drug absorption utilization rate. In terms of preparation technology, the optimized medicinal material extraction method fully retains the efficacy, and the microneedle preparation uses soluble materials. The microneedle quality and performance are ensured through ultrasonic stirring, centrifugation and other processes, and the extract content is accurately controlled. In addition, the main ingredients of the patch are natural medicinal materials, which are highly safe and reduce adverse reactions. The choice of a variety of moisturizers provides flexibility in formula design to meet different needs and has a wide range of applications. DETAILED DESCRIPTION

[0027] In order to better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention.

[0028] Unless otherwise specified, the experimental methods used in the embodiments of the present invention are all conventional methods.

[0029] Unless otherwise specified, the materials, reagents, etc. used in the embodiments of the present invention can be obtained from commercial sources.

[0030] The following introduces the basic information of the main raw materials based on the "Chinese Pharmacopoeia", "Chinese Materia Medica Dictionary", "Chinese Materia Medica", etc.

[0031] Herba pedunculatae: Nature and flavor: warm, pungent. Enters kidney meridian and liver meridian. Functions: dispel wind and dampness, relax muscles and activate blood circulation, disperse blood stasis and reduce swelling, detoxify and relieve pain.

[0032] Elderberry: Nature and flavor: sweet, bitter, neutral. Enters the liver meridian. Functions and indications: bone and tendon repair, blood circulation and pain relief, wind and dampness removal.

[0033] Earthworm: Nature and flavor: salty, cold; slightly toxic. Enters the liver meridian. Functions and indications: dissolves blood stasis, strengthens tendons and bones.

[0034] Stretching grass: Nature and flavor: slightly bitter, pungent, warm. Enters the liver, spleen, and kidney meridians. Functions and indications: dispel wind and dampness, relax muscles and activate collaterals. Used for joint pain and difficulty in flexion and extension.

[0035] Illicium verum: Nature and flavor: warm, bitter and pungent; toxic. The meridians it occupies are not clearly stated, but it is mostly used for liver meridian-related diseases. Functions and indications: clearing away heat and detoxifying, resolving phlegm and dispersing stagnation, removing phlegm and reducing swelling. Used for carbuncle, furuncle, scrofula, sore throat, traumatic injury, and snake bite.

[0036] Drynaria: Nature and flavor: bitter, warm. Enters the kidney and liver meridians. Functions and indications: promotes blood circulation, heals wounds and relieves pain, nourishes the kidney and strengthens bones; externally used to eliminate wind and remove spots.

[0037] Eucommia ulmoides: Nature and flavor: sweet, warm. Enters the liver and kidney meridians. Functions and indications: nourishes the liver and kidneys, strengthens the tendons and bones, and stabilizes pregnancy.

[0038] Achyranthes bidentata: Nature and flavor: bitter, sweet, sour, neutral. Enters the liver and kidney meridians. Functions and indications: nourishes the liver and kidneys, strengthens tendons and bones, removes blood stasis and promotes menstruation, and guides blood downward. Used for soreness of waist and knees, weakness of tendons and bones, amenorrhea, lumps, and dizziness caused by liver yang.

[0039] Chuanxiong: Nature and flavor: pungent, warm. Enters the liver, gallbladder, and pericardium meridians. Functions and indications: promotes blood circulation and qi, dispels wind and relieves pain.

[0040] Safflower: Nature and flavor Meridian: Pungent, warm. Enters the heart and liver meridians. Functions and indications: Activates blood circulation, promotes menstruation, dissipates blood stasis and relieves pain.

[0041] Frankincense: Nature and flavor: pungent, bitter, warm. Enters the heart, liver, and spleen meridians. Functions and indications: Activates blood circulation, promotes qi circulation, relieves pain, reduces swelling, and promotes tissue regeneration.

[0042] Myrrh: Nature and flavor: pungent, bitter, neutral. Enters the heart, liver, and spleen meridians. Functions and indications: dissipates blood stasis and relieves pain, reduces swelling and promotes tissue regeneration.

[0043] Xuejie: ​​Nature and flavor: sweet, salty, neutral. It enters the heart and liver meridians. Functions and indications: promoting blood circulation and relieving pain, removing blood stasis and stopping bleeding, promoting tissue regeneration and healing sores.

[0044] Borneol: Nature and flavor: pungent, bitter, slightly cold. Enters the heart, lung, and liver meridians. Functions and indications: Refreshing the mind, clearing away heat and relieving pain. Example 1

[0045] A topical patch for promoting blood circulation, removing blood stasis, strengthening tendons and strengthening bones, wherein the topical patch is prepared by coating a microneedle-patch composite preparation on a backing material, wherein the microneedle-patch composite preparation is a patch drug with microneedles, and the patch drug comprises the following raw materials in weight percentage: 40% active drug ingredient, 5% sustained-release agent, 2% permeabilizer, 1% absorbent, and the remainder is a moisturizer.

[0046] The active ingredients of the medicine include the following raw materials in parts by weight: 10 parts of Illustris odoratus, 10 parts of frankincense, 10 parts of myrrh, 5 parts of Sanguisorba officinalis, 15 parts of Drynaria fortunei, 15 parts of Eucommia ulmoides, 10 parts of Achyranthes bidentata, 10 parts of Chuanxiong, 5 parts of Carthamus tinctorius, 8 parts of Herba schizonepetae radixe, 10 parts of Elderberry, 6 parts of Earthworm, 10 parts of Herba Lycopodii, and 3 parts of Borneol.

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

[0048] The penetrants are limonene, gingerol and eucalyptol in a mass ratio of 5:4:5.

[0049] The absorbent is a liquid of polyglutamic acid and snail polypeptide in a mass ratio of 0.3:6.

[0050] The snail polypeptide liquid is prepared by placing the snail in a homogenizer, adding deionized water 3 times the mass thereof for homogenization to obtain a snail homogenate, adding a composite protease, performing an enzymatic hydrolysis reaction at a pH of 7.5 and a temperature of 35°C for 2 hours to obtain an enzymatic hydrolyzate, heating the solution to 80°C and maintaining the reaction for 10 minutes to terminate the enzyme reaction, centrifuging, collecting the supernatant, ultrafiltering, and dialyzing to obtain a snail polypeptide liquid with a molecular weight of less than 1kD.

[0051] The moisturizing agent is selected from hesperidin. Example 2

[0052] A topical patch for promoting blood circulation, removing blood stasis, strengthening tendons and strengthening bones, wherein the topical patch is prepared by coating a microneedle-patch composite preparation on a backing material, wherein the microneedle-patch composite preparation is a patch drug with microneedles, and the patch drug comprises the following raw materials in weight percentage: 60% of active pharmaceutical ingredients, 10% of sustained-release agent, 4% of permeabilizing agent, 3% of absorbent, and the remainder is a moisturizing agent.

[0053] The active ingredients of the medicine include the following raw materials in parts by weight: 14 parts of Illustris odoratus, 20 parts of frankincense, 20 parts of myrrh, 15 parts of Sanguisorba officinalis, 30 parts of Drynaria fortunei, 30 parts of Eucommia ulmoides, 20 parts of Achyranthes bidentata, 20 parts of Chuanxiong, 15 parts of Carthamus tinctorius, 16 parts of Herba schizonepetae radixe, 20 parts of Elderberry, 12 parts of Earthworm, 18 parts of Herba Lycopodii, and 8 parts of Borneol.

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

[0055] The penetrant is limonene, gingerol and eucalyptol in a mass ratio of 15:8:10.

[0056] The absorbent is a liquid of polyglutamic acid and snail polypeptide in a mass ratio of 1.5:10.

[0057] The snail polypeptide liquid is prepared by placing the snail in a homogenizer, adding deionized water of 5 times the mass for homogenization to obtain a snail homogenate, adding a composite protease, performing an enzymatic hydrolysis reaction at a pH of 8.5 and a temperature of 40°C for 6 hours to obtain an enzymatic hydrolyzate, heating to 90°C, maintaining for 15 minutes, terminating the enzyme reaction, centrifuging, collecting the supernatant, ultrafiltering, and dialyzing to obtain a snail polypeptide liquid with a molecular weight of less than 1kD.

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

[0059] A topical patch for promoting blood circulation, removing blood stasis, strengthening tendons and strengthening bones, wherein the topical patch is prepared by coating a microneedle-patch composite preparation on a backing material, wherein the microneedle-patch composite preparation is a patch drug with microneedles, and the patch drug comprises the following raw materials in weight percentage: 50% active drug ingredient, 8% sustained-release agent, 3% permeabilizer, 2% absorbent, and the remainder is a moisturizer.

[0060] The active ingredients of the medicine include the following raw materials in parts by weight: 12 parts of Illustris odoratus, 15 parts of frankincense, 15 parts of myrrh, 10 parts of Sanguisorba officinalis, 22 parts of Drynaria fortunei, 28 parts of Eucommia ulmoides, 15 parts of Achyranthes bidentata, 15 parts of Chuanxiong, 10 parts of Carthamus tinctorius, 12 parts of Herba schizonepetae radixe, 15 parts of Sambucus sambucus, 9 parts of Earthworm, 15 parts of Herba Lycopodii, and 5 parts of Borneol.

[0061] The sustained-release agent is cyclodextrin, tragacanth gum and locust bean gum in a mass ratio of 20:6:8.

[0062] The penetrant is limonene, gingerol and eucalyptol in a mass ratio of 10:6:8.

[0063] The absorbent is a liquid of polyglutamic acid and snail polypeptide in a mass ratio of 0.9:8.

[0064] The snail polypeptide liquid is prepared by placing the snail in a homogenizer, adding deionized water of 4 times the mass for homogenization to obtain a snail homogenate, adding a composite protease, performing an enzymatic hydrolysis reaction at a pH of 8 and a temperature of 38° C. for 4 hours to obtain an enzymatic hydrolyzate, heating to 85° C., maintaining for 12 minutes, terminating the enzyme reaction, centrifuging, collecting the supernatant, ultrafiltering, and dialyzing to obtain a snail polypeptide liquid with a molecular weight of less than 1kD.

[0065] The moisturizing agent is selected from vaseline.

[0066] The above examples 1-3 adopt the following preparation process:

[0067] S1. Wash and dry the Radix Angelicae Pubescentis, Radix Elderberry, Radix Glehniae, Herba Lycopodii, Rhizoma Drynariae, Eucommia Bark, Rhizoma Cyathulae, Rhizoma Chuanxiong, and Flos Carthami, grind them into coarse powder, and sieve through a 70-mesh sieve for later use; grind frankincense, myrrh, Sanguisorba Officinalis, and borneol into fine powder, and sieve through a 100-mesh sieve for later use;

[0068] S2. Add 80% ethanol solution to the above medicinal materials, let them stand and soak for 8-12 hours, heat to 70°C and extract 4 times, each time for 4 hours, and combine the extracts;

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

[0070] S4, dissolving polylactic acid-glycolic acid copolymer in dichloromethane with a mass volume g / mL ratio of 1:8, stirring evenly to form a uniform solution, then adding the extract of S3, and then ultrasonically stirring for 3 hours, with an ultrasonic power of 700 W, an ultrasonic frequency of 30 kHz, and a stirring rate of 600 rpm to obtain a mixed solution, and slowly injecting the mixed solution into the microneedle mold, with the extract content accounting for 20% of the total mass of the microneedle, centrifuging at a rate of 4000 rpm for 5-10 min, and then vacuum drying at a vacuum degree of -0.1 MPa and a temperature of 35°C, and demolding to obtain a microneedle-patch composite preparation;

[0071] S5. Apply the above-mentioned microneedle-patch composite preparation on a backing material to obtain an external patch.

[0072] Comparative Example 1

[0073] The difference between this comparative example and Example 3 is that the external plaster is a plaster drug directly coated on the backing material.

[0074] Comparative Example 2

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

[0076] Comparative Example 3

[0077] The difference between this comparative example and Example 3 is that the active pharmaceutical ingredient of the patch 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 pharmaceutical ingredient of the patch does not contain a penetrant.

[0080] Test Example 1 - In vitro transdermal test

[0081] 1. Skin preparation: Remove subcutaneous fat from fresh ex vivo pig skin, rinse with saline, and then dry 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 fits tightly and without leakage.

[0082] 2. Plaster placement: accurately cut the test patches of Examples 1-3 and Comparative Examples 1-4 into 2.5 cm, and evenly apply them on the skin surface to make them fully contact with the skin.

[0083] 3. Receiving solution replacement and sampling: Add 5 mL of receiving solution to the receiving pool, maintain the temperature at 37°C ± 0.5°C, and stir at a constant speed (500 rpm). Take out 0.5 mL of receiving solution from the receiving pool at 0.5, 1, 2, 4, 6, 8, 12, and 24 hours, and add an equal amount of fresh receiving solution to maintain a constant volume of receiving solution.

[0084] 4. Determination of drug content: After the taken receptor solution is centrifuged and filtered, the drug content therein is determined by high performance liquid chromatography. The cumulative amount of drug permeated through the skin at different time points is calculated according to the standard curve, and the drug permeation rate is calculated.

[0085] 5. Test results

[0086]

[0087] Test Example 2-Pharmacodynamics Experiment

[0088] 1. Experimental Animals

[0089] Select healthy adult rats weighing 180-220 g (rats), half male and half female.

[0090] 2. Disease model establishment

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

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

[0093] 3. Grouping and Dosing

[0094] The animals were randomly divided into an experimental group (patch administration group), a positive control group (commercially available 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 plasters of Examples 1-3 and Comparative Examples 1-4 were placed in a 3*3 cm 2 The area and 400 mg dose were applied to the fracture site of the animal and changed once a day.

[0096] Positive control group: The commercially available Yunnan Baiyao ointment was administered in the same manner and at the same dosage.

[0097] Negative control group: blank patch was applied, and the operation was the same as that of the experimental group.

[0098] 4. Observation indicators

[0099] (1) Fracture model observation indicators

[0100] Imaging examination: On the 7th, 14th and 21st days after administration, animals were subjected to X-rays to observe fracture healing and measure indicators such as the area of ​​callus formation and the degree of fracture line healing.

[0101] Biomechanical testing: 28 days after administration, biomechanical testing was performed on the animals' fractured limbs to measure parameters such as the maximum load and elastic modulus of the bones and to evaluate the recovery of the mechanical properties of the bones.

[0102] (2) Observation indicators of arthritis model

[0103] Joint swelling: Use a vernier caliper to measure the circumference of the animal's joints 2-3 times a week and calculate the joint swelling to assess the degree of joint inflammation.

[0104] Pain behavior test: Use the hot plate method or tenderness method to regularly measure the pain threshold of the animals, observe the changes in the animals' pain behavior, and evaluate the analgesic effect of the patch.

[0105] Histopathological examination: After the experiment, the joint tissues of the animals were taken for pathological sections to observe the lesions of tissues such as articular cartilage and synovium, and the degree of 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 shows that the example plasters can more effectively promote the early formation of callus, among which Example 3 has the best effect.

[0110] On the 28th day, the fracture line of Example 3 completely disappeared and basically healed. Examples 1 and 2 also healed significantly, with the fracture line close to disappearing or almost invisible. The fracture line of the comparative group was more blurred and healed better, but still not as good as the example group. The fracture line of the positive control group was still visible, and the degree of healing was average. The fracture line of the negative control group was clear and healed slowly.

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

[0112] Arthritis Model:

[0113]

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

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

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

[0117] Test Example 3 - Clinical Trial

[0118] 1. Experimental Design

[0119] A randomized, double-blind, parallel-controlled clinical trial design was adopted.

[0120] 2. Subject Selection

[0121] Patients who met the following criteria were recruited

[0122] (1) Applicants must be between 18 and 65 years old, regardless of gender.

[0123] (2) Patients suffering from fractures, traumatic injuries, osteoarthritis, rheumatoid arthritis and other diseases that fall within the scope of treatment for promoting blood circulation and removing blood stasis, strengthening tendons and bones, and whose condition is in a stable stage.

[0124] (3) Patients voluntarily participated in the trial and signed the informed consent form.

[0125] Exclusion criteria

[0126] (1) Those who are allergic to any ingredient in the patch.

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

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

[0129] 3. Grouping and Dosing

[0130] The subjects were randomly divided into an experimental group and a 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 to the patients. 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 mobility were scored. The visual analogue scale (VAS) was used to assess the pain severity, and the swelling and limited joint mobility 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] After analyzing the pain VAS score, the improvement rate of joint movement limitation and the improvement rate of imaging in this clinical trial, it was found that the pain VAS scores of each experimental group, comparative group, positive control group and negative control group before treatment were similar. After treatment, the pain VAS scores of all groups decreased, among which the embodiment group performed outstandingly in terms of pain relief, improvement of joint movement limitation and imaging improvement, especially embodiment 3 had the best effect, with a pain score of only 1.5±0.3 after 4 weeks of treatment, an improvement rate of joint movement limitation of 85.6%±8.3%, and an imaging improvement rate of 78.6%±7.5%.

[0140] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. An external plaster for promoting blood circulation, removing blood stasis, strengthening tendons and strengthening bones, characterized in that: 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 comprises the following raw materials in weight percentage: 40-60% active drug ingredients, 5-10% sustained-release agent, 2-4% permeabilizer, 1-3% absorbent, and the remainder is a moisturizer; the active drug ingredients comprise the following raw materials in weight percentage: 10-14 parts of octagonal lotus, 10 parts of frankincense, -20 parts, myrrh 10-20 parts, blood clotting 5-15 parts, drynariae rhizome 15-30 parts, eucommia ulmoides 15-30 parts, Achyranthes bidentata 10-20 parts, ligusticum chuanxiong 10-20 parts, safflower 5-15 parts, radix schizonepetae 8-16 parts, elderberry 10-20 parts, earthworm 6-12 parts, lycopodiella cuneata 10-18 parts, borneol 3-8 parts, the absorbent is a polyglutamic acid and snail polypeptide liquid in a mass ratio of (0.3-1.5):(6-10).

2. The external plaster for promoting blood circulation and removing blood stasis, strengthening tendons and strengthening bones as claimed in claim 1, characterized in that: The plaster medicine is prepared from the following raw materials in weight percentage: 40-60% of active pharmaceutical ingredients, 5-10% of sustained-release agent, 2-4% of permeation agent, 1-3% of absorbent, and the balance of moisturizing agent.

3. The external plaster for promoting blood circulation and removing blood stasis, strengthening tendons and strengthening bones as claimed in claim 1, characterized in that: The active ingredients of the medicine include the following raw materials in parts by weight: 12 parts of Illustris odoratus, 15 parts of frankincense, 15 parts of myrrh, 10 parts of Sanguisorba officinalis, 22 parts of Drynaria fortunei, 28 parts of Eucommia ulmoides, 15 parts of Achyranthes bidentata, 15 parts of Chuanxiong, 10 parts of Carthamus tinctorius, 12 parts of Herba schizonepetae radixe, 15 parts of Sambucus sambucus, 9 parts of Earthworm, 15 parts of Herba Lycopodii, and 5 parts of Borneol.

4. The external plaster for promoting blood circulation and removing blood stasis, strengthening tendons and strengthening bones as claimed in claim 1, characterized in that: The sustained-release agent is cyclodextrin, tragacanth gum and locust bean gum in a mass ratio of (12-25):(4-8):(5-10).

5. The external plaster for promoting blood circulation and removing blood stasis, strengthening tendons and strengthening bones as claimed in claim 1, characterized in that: The penetrants are limonene, gingerol and eucalyptol in a mass ratio of (5-15):(4-8):(5-10).

6. The external plaster for promoting blood circulation and removing blood stasis, strengthening tendons and strengthening bones as claimed in 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., maintaining the reaction for 10-15 minutes, terminating the enzyme reaction, centrifuging, collecting the supernatant, ultrafiltering, and dialyzing to obtain a snail polypeptide liquid with a molecular weight of less than 1kD.

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

8. The preparation process of the external plaster for promoting blood circulation and 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 Radix Angelicae Pubescentis, Radix Elderberry, Radix Glehniae, Herba Lycopodii, Rhizoma Drynariae, Eucommia Bark, Rhizoma Cyathulae, Rhizoma Chuanxiong, and Flos Carthami, grind them into coarse powder, and sieve through 60-80 mesh for later use; grind frankincense, myrrh, and dragon's blood into fine powder, and sieve through 100-120 mesh for later use; S2. Add 70-90% ethanol solution to the above medicinal materials, soak for 8-12 hours, heat to 50-80°C, extract 3-5 times, each time for 2-4 hours, and combine the extracts; S3, concentrate the extract at 60-80℃ and reduced pressure -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 with a mass volume g / mL ratio of 1:5-10, stirring evenly to form a uniform solution, then adding the extract of S3, and then ultrasonically stirring for 2-5h to obtain a mixed solution, slowly injecting the mixed solution into the microneedle mold, the extract content accounts for 10-30% of the total mass of the microneedle, centrifuging at a rate of 3000-5000rpm for 5-10min, and then vacuum drying at a vacuum degree of -0.08~-0.1MPa and a temperature of 30-40°C, demolding, and obtaining a microneedle-patch composite preparation; S5. Apply the above-mentioned microneedle-patch composite preparation on a backing material to obtain an external patch.

9. The preparation process of the external plaster for promoting blood circulation, removing blood stasis, strengthening tendons and strengthening bones as claimed in claim 8, characterized in that: The soluble material of S4 is selected from polylactic acid-glycolic acid copolymer, hyaluronic acid or chitosan.

10. The preparation process of the external plaster for promoting blood circulation, removing blood stasis, strengthening tendons and strengthening bones as claimed in claim 8, 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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