Preparation and application method of a composition for treating neck, shoulder, waist and leg pain

Abstract

The present invention provides a preparation method and application method of a composition for treating neck, shoulder, waist and leg pain, belonging to the field of pharmaceutical technology; the preparation method comprises the following steps: preparing a mixed Chinese medicine extract; preparing a nanosilver composite solution; preparing a composite matrix solution; and preparing a Chinese medicine composition microneedle. The present invention prepares a composite matrix solution by mixing degradable and highly biocompatible hyaluronic acid and chitosan, and then uses the composite matrix solution and the mixed Chinese medicine extract as raw materials to prepare the Chinese medicine composition microneedle. Since the Chinese medicine composition microneedle is composed of a swellable polymer cross-linked hydrogel, after the needle tip pierces the skin, a microporous channel is left, so that the mixed Chinese medicine extract loaded by it absorbs interstitial fluid through the microporous channel with the carrier material, thereby expanding, so that the drug is continuously released from the carrier material to the site of action, thereby achieving the effect of improving the utilization efficiency of the drug.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to a preparation method of a composition for treating neck, shoulder, waist and leg pain and an application method thereof. Background Art

[0002] Neck, shoulder, waist and leg pain refers to a series of pain symptoms occurring in the neck, shoulders, waist and legs, which are characterized by recurring attacks. There are many causes of neck, shoulder, waist and leg pain, which are related to soft tissue injury, chronic strain of soft tissue, wind, cold and dampness, allergies or immune factors. Clinical symptoms include low back pain, intermittent claudication, radiating pain in one or both lower limbs, numbness of the lower limbs and abnormal sensation.

[0003] At present, there are a variety of traditional Chinese medicine preparations for the treatment of neck, shoulder, waist and leg pain, which are mainly divided into oral traditional Chinese medicine preparations and topical preparations. Oral preparations are inconvenient to take, carry and make, and usually have problems such as poor patient compliance and first-pass effect of drugs. Although topical preparations have good compliance, strong targeting, can avoid first-pass effect and reduce systemic side effects, they usually have low penetration rate due to drug accumulation in the stratum corneum, resulting in low drug utilization rate and unable to quickly and effectively exert therapeutic effects.

[0004] Therefore, it is necessary to propose a composition for treating neck, shoulder, waist and leg pain and an application method thereof which can improve the utilization rate of the drug, so as to improve the therapeutic effect. Summary of the invention

[0005] In view of the shortcomings of the prior art, the object of the present invention is to provide a preparation method of a composition for treating neck, shoulder, waist and leg pain and an application method thereof.

[0006] A method for preparing a composition for treating neck, shoulder, waist and leg pain comprises the following steps:

[0007] S1: Preparation of mixed Chinese herbal medicine extracts

[0008] Mixing and grinding the herb and gentiana macrophylla, and fully mixing them with distilled water, then adding cellulase for enzymolysis to prepare an enzymolysis solution, and then mixing and grinding chuanxiong, angelica dahurica, cassia twig, clematis root, safflower, achyranthes bidentata, millettia spatholobi and asarum, and mixing them with the ethanol solution and the enzymolysis solution, and performing microwave-assisted heating reflux extraction to obtain a mixed Chinese medicine extract;

[0009] S2: Preparation of nanosilver composite solution

[0010] Dissolving chitosan and tannic acid in acetic acid solution and distilled water respectively to prepare chitosan-acetic acid solution and tannic acid solution, then stirring and mixing the tannic acid solution and the chitosan-acetic acid solution, adding silver nitrate aqueous solution, stirring and reacting to obtain a nano-silver composite solution;

[0011] S3: Preparation of composite matrix solution

[0012] Add hyaluronic acid into distilled water at a solid-liquid ratio of 1 g: (20-30) mL, stir until completely dissolved, then add the above-mentioned nanosilver composite solution, stir and mix evenly to obtain a composite matrix solution;

[0013] S4: Preparation of microneedles of Chinese medicine composition

[0014] The above-mentioned composite matrix solution is poured into a microneedle mold, and after centrifugation, freezing and thawing, a microneedle needle head mold is made. Then, the above-mentioned mixed Chinese medicine extract is added to the microneedle needle head mold, flattened and the composite matrix solution is added again. After centrifugation, freezing and thawing, it is dried and demolded to obtain a Chinese medicine composition microneedle.

[0015] Furthermore, S1 specifically includes the following steps:

[0016] S1.1: Mix and grind the herb Loranthus parasiticus and Gentiana macrophylla, add them into distilled water at a solid-liquid ratio of 1 g: (30-40) mL, heat and stir at 40-50°C for 1-2 h to obtain a binary mixed suspension;

[0017] S1.2: Add 0.3-0.5 wt% cellulase to the above binary mixed suspension, keep warm for 40-50 min, then heat to 90-100°C to inactivate the enzyme for 3-5 min to obtain an enzymatic solution;

[0018] S1.3: Mix and grind Chuanxiong, Angelica dahurica, Cinnamomum cassia, Clematis chinensis, Carthamus tinctorius, Achyranthes bidentata, Millettia spatholobi and Asarum, add them into 60% ethanol solution at a solid-liquid ratio of 1g:(40-50)mL, then add the above enzymatic solution, stir and mix evenly to obtain a composite suspension;

[0019] S1.4: placing the composite suspension in a microwave extractor, performing microwave-assisted heating reflux extraction for 2 to 3 hours at a temperature of 80 to 90°C and a power of 80 to 100 W, and filtering to obtain an extract;

[0020] S1.5: Place the above extract in a concentrator, heat and concentrate it, remove the solvent, and obtain a mixed Chinese medicine extract.

[0021] Furthermore, S2 specifically includes the following steps:

[0022] S2.1: Add chitosan to acetic acid solution at a solid-liquid ratio of 1 g: (20-30) mL, stir thoroughly to dissolve, and prepare chitosan-acetic acid solution;

[0023] S2.2: Add tannic acid to distilled water, heat and stir at 70-90°C until fully dissolved, to obtain a 200-300 μg / mL tannic acid solution;

[0024] S2.3: Add the above tannic acid solution to the above chitosan-acetic acid solution in a volume ratio of 1:(3-4), stir and mix evenly, then add the silver nitrate aqueous solution, continue stirring for 2-3 hours, react and obtain a nanosilver composite solution.

[0025] Furthermore, S4 specifically includes the following steps:

[0026] S4.1: Pour the composite matrix solution prepared in step S3 into the microneedle mold, place it in a centrifuge, centrifuge it horizontally at a rate of 3000-4000 r / min for 4-6 minutes, then rotate it 180 degrees, continue centrifugation for 4-6 minutes, repeat 2-3 times, take it out, place it in a freezer, freeze it at -50--40°C for 1-2 hours, and finally thaw it in a medium-heat microwave for 20-30 seconds to obtain a microneedle needle mold;

[0027] S4.2: Add the mixed Chinese medicine extract obtained in step S1.5 to the above-mentioned microneedle needle mold, flatten it, add the composite matrix solution, centrifuge at a rate of 3000-4000 r / min for 4-6 minutes, then rotate 180 degrees, continue centrifugation for 4-6 minutes, repeat 3-5 times, and obtain a microneedle backing mold;

[0028] S4.3: Place the above-mentioned microneedle backing mold in a freezer and freeze it at -50 to -40°C for 1 to 2 hours. After taking it out, thaw it in a microwave at medium heat for 20 to 30 seconds. Repeat the freeze-thaw process 3 to 4 times, dry it at room temperature for 20 to 24 hours and demold it to obtain the traditional Chinese medicine composition microneedles.

[0029] Furthermore, the mixed Chinese medicine extract raw material components include, by weight: 20 to 30 parts of Chuanxiong, 12 to 18 parts of Angelica dahurica, 12 to 16 parts of cinnamon twig, 8 to 14 parts of Viscum album, 6 to 10 parts of Clematis chinensis, 7 to 9 parts of Carthamus tinctorius, 3 to 5 parts of Achyranthes bidentata, 3 to 5 parts of Millettia reticulata, 3 to 5 parts of Asarum and 1 to 3 parts of Gentiana macrophylla.

[0030] Furthermore, the volume fraction of the acetic acid solution is 2-4%.

[0031] Furthermore, the volume ratio of the silver nitrate aqueous solution to the tannic acid solution is 1:(2-3), and the concentration of the silver nitrate aqueous solution is 10-20 μg / mL.

[0032] Furthermore, the mass ratio of hyaluronic acid to nano-silver composite solution chitosan is (3-5):1.

[0033] Furthermore, the mass ratio of the mixed Chinese herbal medicine extract to the composite matrix solution is 1:(400-500).

[0034] A method for preparing a composition for treating cervical, shoulder, waist and leg pains, and application of the prepared composition in preparing a medicine for treating cervical, shoulder, waist and leg pains.

[0035] Compared with the prior art, the present invention has the following beneficial effects:

[0036] 1. The present invention prepares a composite matrix solution by mixing degradable and highly biocompatible hyaluronic acid and chitosan, and then uses the composite matrix solution and a mixed Chinese medicine extract as raw materials to prepare a Chinese medicine composition microneedle. Since the Chinese medicine composition microneedle is composed of a swellable polymer cross-linked hydrogel, the needle tip pierces the skin, leaving a microporous channel, so that the mixed Chinese medicine extract loaded by it absorbs interstitial fluid through the microporous channel along with the carrier material, thereby expanding, so that the drug is continuously released from the carrier material to the site of action, thereby achieving the effect of improving the utilization efficiency of the drug.

[0037] 2. The present invention dissolves chitosan in an acetic acid solution and then stirs and mixes the solution with an aqueous solution of tannic acid. Due to the hydrogen bonding between the amino groups of chitosan and the hydroxyl groups of tannic acid, the tannic acid is fixed in the chitosan support to form a gel network. Then, an aqueous silver nitrate solution is added to reduce anions using the phenolic hydroxyl groups of the tannic acid molecules to generate nano-silver with antibacterial effect. Since the amino groups of chitosan are positively charged, they can adsorb negatively charged bacteria. Therefore, when the chitosan and nano-silver are compounded, the antibacterial effect can be further improved.

[0038] 3. The present invention comprises the following steps: firstly mixing and crushing Herba Mori and Radix Gentianae, mixing them with distilled water, adding cellulase for enzymolysis to prepare an enzymolysis solution, then stirring and mixing the enzymolysis solution and ethanol solution with mixed powder of crushed and ground Rhizoma Chuanxiong, Rhizoma Angelicae Pubescentis, Rhizoma Cinnamomi, Radix Clematis, Carthamus tinctorius, Rhizoma Achyranthis Bidentatae, Caulis Spatholobi and Asarum, and performing microwave-assisted heating reflux extraction to prepare a mixed Chinese medicine extract with the effects of promoting blood circulation and removing blood stasis, dispelling wind and dredge meridians, and having anti-inflammatory and analgesic effects, which can effectively treat pain in the neck, shoulder, waist and legs, wherein Rhizoma Cinnamomi and Asarum have a synergistic effect, and adding the two simultaneously can further improve the efficacy of the mixed Chinese medicine composition. DETAILED DESCRIPTION

[0039] The following is a detailed description of the preparation of a composition for treating neck, shoulder, waist and leg pain and its application method provided by the present invention in conjunction with specific embodiments. At the same time, it is explained here that in order to make the embodiments more detailed, the following embodiments are the best and preferred embodiments, and those skilled in the art may also adopt other alternatives to implement some known technologies.

[0040] Example 1

[0041] A method for preparing a composition for treating neck, shoulder, waist and leg pain comprises the following steps:

[0042] S1: Preparation of mixed Chinese herbal medicine extracts

[0043] S1.1: Mix and grind the herb Loranthus parasiticus and Gentiana macrophylla, add them into distilled water at a solid-liquid ratio of 1 g:30 mL, heat and stir at 40 °C for 1 h to obtain a binary mixed suspension;

[0044] S1.2: adding 0.3 wt% cellulase to the binary mixed suspension, preserving the mixture for enzymolysis for 40 min, then heating the mixture to 90°C for 3 min to inactivate the enzyme, and obtaining an enzymolysis solution;

[0045] S1.3: Mix and grind Rhizoma Chuanxiong, Angelica dahurica, Cinnamomum cassia, Clemati radix, Carthamus tinctorius, Achyranthes bidentata, Millettia spatholobi and Asarum, add them into 60% ethanol solution at a solid-liquid ratio of 1g:40mL, then add the above enzymatic solution, stir and mix evenly to obtain a composite suspension;

[0046] S1.4: placing the composite suspension in a microwave extractor, performing microwave-assisted heating reflux extraction for 2 h at a temperature of 80° C. and a power of 80 W, and filtering to obtain an extract;

[0047] S1.5: placing the extract in a concentrator, heating and concentrating, removing the solvent, and obtaining a mixed Chinese medicine extract;

[0048] The raw material components of the mixed Chinese medicine extract include, by weight: 20 parts of Chuanxiong, 12 parts of Angelica dahurica, 12 parts of Cinnamomum cassia, 8 parts of Morus alba, 6 parts of Clematis, 7 parts of Carthamus tinctorius, 3 parts of Achyranthes bidentata, 3 parts of Millettia reticulata, 3 parts of Asarum and 1 part of Gentiana macrophylla;

[0049] S2: Preparation of nanosilver composite solution

[0050] S2.1: chitosan is added to acetic acid solution at a solid-liquid ratio of 1 g:20 mL, and the mixture is stirred and dissolved to prepare chitosan-acetic acid solution, wherein the volume fraction of the acetic acid solution is 2%;

[0051] S2.2: Add tannic acid to distilled water, heat and stir at 70°C until fully dissolved, to obtain a 200 μg / mL tannic acid solution;

[0052] S2.3: adding the tannic acid solution to the chitosan-acetic acid solution at a volume ratio of 1:3, stirring and mixing evenly, then adding a silver nitrate aqueous solution, continuing stirring for 2 hours, reacting to obtain a nanosilver composite solution, wherein the volume ratio of the silver nitrate aqueous solution to the tannic acid solution is 1:2, and the concentration of the silver nitrate aqueous solution is 10 μg / mL;

[0053] S3: Preparation of composite matrix solution

[0054] Add hyaluronic acid into distilled water at a solid-liquid ratio of 1 g: 20 mL, stir until completely dissolved, then add the above nano-silver composite solution, stir and mix evenly to obtain a composite matrix solution, wherein the mass ratio of hyaluronic acid to chitosan in the nano-silver composite solution is 3:1;

[0055] S4: Preparation of microneedles of Chinese medicine composition

[0056] S4.1: Pour the composite matrix solution prepared in step S3 into the microneedle mold, place it in a centrifuge, centrifuge it horizontally at a rate of 3000r / min for 4min, then rotate it 180°, continue centrifugation for 4min, repeat twice, take it out, place it in a freezer, freeze it at -40℃ for 1h, and finally thaw it in a microwave at medium heat for 20s to obtain a microneedle needle mold;

[0057] S4.2: Add the mixed Chinese herbal extract obtained in step S1.5 to the above-mentioned microneedle needle mold, flatten it, add the composite matrix solution, centrifuge at a rate of 3000r / min for 4min, then rotate 180°, continue centrifugation for 4min, repeat 3 times to obtain a microneedle backing mold, wherein the mass ratio of the mixed Chinese herbal extract to the composite matrix solution is 1:400;

[0058] S4.3: Place the above-mentioned microneedle backing mold in a freezer and freeze it at -40°C for 1 hour. After taking it out, thaw it in a microwave at medium heat for 20 seconds. Repeat the freeze-thaw process three times, dry it at room temperature for 20 hours and demold it to obtain the traditional Chinese medicine composition microneedle.

[0059] Example 2

[0060] A method for preparing a composition for treating neck, shoulder, waist and leg pain comprises the following steps:

[0061] S1: Preparation of mixed Chinese herbal medicine extracts

[0062] S1.1: Mix and grind the Herba Mori and Radix Gentianae, add them into distilled water at a solid-liquid ratio of 1 g:35 mL, heat and stir at 45 °C for 1.5 h to obtain a binary mixed suspension;

[0063] S1.2: adding 0.4 wt% cellulase to the binary mixed suspension, preserving the mixture for 45 min, and then heating the mixture to 95°C to inactivate the enzyme for 4 min to obtain an enzymatic solution;

[0064] S1.3: Mix and grind Rhizoma Chuanxiong, Angelica dahurica, Cinnamomum cassia, Clemati radix, Carthamus tinctorius, Achyranthes bidentata, Millettia spatholobi and Asarum, add them into 60% ethanol solution at a solid-liquid ratio of 1g:45mL, then add the above enzymatic solution, stir and mix evenly to obtain a composite suspension;

[0065] S1.4: placing the composite suspension in a microwave extractor, performing microwave-assisted heating reflux extraction for 2.5 h at a temperature of 85° C. and a power of 90 W, and filtering to obtain an extract;

[0066] S1.5: placing the extract in a concentrator, heating and concentrating, removing the solvent, and obtaining a mixed Chinese medicine extract;

[0067] The raw material components of the mixed Chinese medicine extract include, by weight: 25 parts of Chuanxiong, 15 parts of Angelica dahurica, 14 parts of Cinnamomum cassia, 11 parts of Morus alba, 8 parts of Clematis, 8 parts of Carthamus tinctorius, 4 parts of Achyranthes bidentata, 4 parts of Millettia reticulata, 4 parts of Asarum and 2 parts of Gentiana macrophylla;

[0068] S2: Preparation of nanosilver composite solution

[0069] S2.1: chitosan is added to acetic acid solution at a solid-liquid ratio of 1 g:25 mL, and the mixture is stirred and dissolved to prepare chitosan-acetic acid solution, wherein the volume fraction of the acetic acid solution is 3%;

[0070] S2.2: Add tannic acid to distilled water, heat and stir at 80°C until fully dissolved, to obtain a 250 μg / mL tannic acid solution;

[0071] S2.3: adding the tannic acid solution to the chitosan-acetic acid solution at a volume ratio of 1:3.5, stirring and mixing evenly, then adding a silver nitrate aqueous solution, continuing stirring for 2.5 hours, reacting to obtain a nanosilver composite solution, wherein the volume ratio of the silver nitrate aqueous solution to the tannic acid solution is 1:2.5, and the concentration of the silver nitrate aqueous solution is 15 μg / mL;

[0072] S3: Preparation of composite matrix solution

[0073] Add hyaluronic acid into distilled water at a solid-liquid ratio of 1 g: 25 mL, stir until completely dissolved, then add the above-mentioned nano-silver composite solution, stir and mix evenly to obtain a composite matrix solution, wherein the mass ratio of hyaluronic acid to chitosan in the nano-silver composite solution is 4:1;

[0074] S4: Preparation of microneedles of Chinese medicine composition

[0075] S4.1: Pour the composite matrix solution prepared in step S3 into the microneedle mold, place it in a centrifuge, centrifuge it horizontally at a rate of 3500r / min for 5min, then rotate it 180°, continue centrifugation for 5min, repeat twice, take it out, place it in a freezer, freeze it at -45℃ for 1.5h, and finally thaw it in a microwave at medium heat for 25s to obtain a microneedle needle mold;

[0076] S4.2: Add the mixed Chinese herbal extract obtained in step S1.5 to the above-mentioned microneedle needle mold, flatten it, add the composite matrix solution, centrifuge at a rate of 3500r / min for 5min, then rotate 180°, continue centrifugation for 5min, repeat 4 times to obtain a microneedle backing mold, wherein the mass ratio of the mixed Chinese herbal extract to the composite matrix solution is 1:450;

[0077] S4.3: Place the above-mentioned microneedle backing mold in a freezer and freeze it at -45°C for 1.5 hours. After taking it out, thaw it in a microwave at medium heat for 25 seconds. Repeat the freeze-thaw process three times, dry it at room temperature for 22 hours and demold it to obtain the traditional Chinese medicine composition microneedles.

[0078] Example 3

[0079] A method for preparing a composition for treating neck, shoulder, waist and leg pain comprises the following steps:

[0080] S1: Preparation of mixed Chinese herbal medicine extracts

[0081] S1.1: Mix and grind the herb Loranthus parasiticus and Gentiana macrophylla, add them into distilled water at a solid-liquid ratio of 1 g:40 mL, heat and stir at 50 °C for 2 h to obtain a binary mixed suspension;

[0082] S1.2: Add 0.5 wt% cellulase to the above binary mixed suspension, keep warm for 50 min for enzymatic hydrolysis, then heat to 100°C for 5 min to inactivate the enzyme, and obtain an enzymatic hydrolyzate;

[0083] S1.3: Mix and grind Rhizoma Chuanxiong, Angelica dahurica, Cinnamomum cassia, Clemati radix, Carthamus tinctorius, Achyranthes bidentata, Millettia spatholobi and Asarum, add them into 60% ethanol solution at a solid-liquid ratio of 1g:50mL, then add the above enzymatic solution, stir and mix evenly to obtain a composite suspension;

[0084] S1.4: placing the composite suspension in a microwave extractor, performing microwave-assisted heating reflux extraction for 3 h at a temperature of 90° C. and a power of 100 W, and filtering to obtain an extract;

[0085] S1.5: placing the extract in a concentrator, heating and concentrating, removing the solvent, and obtaining a mixed Chinese medicine extract;

[0086] The raw material components of the mixed Chinese medicine extract include, by weight: 30 parts of Chuanxiong, 18 parts of Angelica dahurica, 16 parts of Cinnamomum cassia, 14 parts of Morus alba, 10 parts of Clematis, 9 parts of Carthamus tinctorius, 5 parts of Achyranthes bidentata, 5 parts of Millettia reticulata, 5 parts of Asarum and 3 parts of Gentiana macrophylla;

[0087] S2: Preparation of nanosilver composite solution

[0088] S2.1: chitosan is added to acetic acid solution at a solid-liquid ratio of 1 g:30 mL, and the mixture is stirred and dissolved to prepare chitosan-acetic acid solution, wherein the volume fraction of the acetic acid solution is 4%;

[0089] S2.2: Add tannic acid to distilled water, heat and stir at 90°C until fully dissolved, to obtain a 300 μg / mL tannic acid solution;

[0090] S2.3: adding the tannic acid solution to the chitosan-acetic acid solution at a volume ratio of 1:4, stirring and mixing evenly, then adding a silver nitrate aqueous solution, continuing stirring for 3 hours, reacting to obtain a nanosilver composite solution, wherein the volume ratio of the silver nitrate aqueous solution to the tannic acid solution is 1:3, and the concentration of the silver nitrate aqueous solution is 20 μg / mL;

[0091] S3: Preparation of composite matrix solution

[0092] Add hyaluronic acid into distilled water at a solid-liquid ratio of 1 g:30 mL, stir until completely dissolved, then add the above-mentioned nano-silver composite solution, stir and mix evenly to obtain a composite matrix solution, wherein the mass ratio of hyaluronic acid to chitosan in the nano-silver composite solution is 5:1;

[0093] S4: Preparation of microneedles of Chinese medicine composition

[0094] S4.1: Pour the composite matrix solution prepared in step S3 into the microneedle mold, place it in a centrifuge, centrifuge it horizontally at a rate of 4000r / min for 6min, then rotate it 180°, continue centrifugation for 6min, repeat 3 times, take it out, place it in a freezer, freeze it at -50℃ for 2h, and finally thaw it in a medium-heat microwave for 30s to obtain a microneedle needle mold;

[0095] S4.2: Add the mixed Chinese herbal extract obtained in step S1.5 to the above-mentioned microneedle needle mold, flatten it, add the composite matrix solution, centrifuge at a rate of 4000r / min for 6min, then rotate 180°, continue centrifugation for 6min, repeat 5 times, and obtain a microneedle backing mold, wherein the mass ratio of the mixed Chinese herbal extract to the composite matrix solution is 1:500;

[0096] S4.3: Place the above-mentioned microneedle backing mold in a freezer and freeze it at -50°C for 2 hours. After taking it out, thaw it in a microwave at medium heat for 30 seconds. Repeat the freeze-thaw process 4 times, dry it at room temperature for 24 hours and demold it to obtain the traditional Chinese medicine composition microneedle.

[0097] Comparative Example 1

[0098] The difference between this comparative example and Example 1 is that steps S2-S4 are removed, and the mixed Chinese herbal extract obtained in S1 is made into a gel paste according to the method of patent CN109432179A, wherein the amount of the mixed Chinese herbal extract added is equal to the amount of the mixed Chinese herbal extract added in step S4 of Example 1.

[0099] Comparative Example 2

[0100] This comparative example is substantially the same as Example 1, except that step S2 is removed, and the nanosilver composite solution in step S3 is replaced by an equal amount of chitosan-acetic acid solution.

[0101] Comparative Example 3

[0102] This comparative example is basically the same as Example 1, except that the silver nitrate aqueous solution in step S2 is replaced by an equal amount of chitosan-acetic acid solution.

[0103] Comparative Example 4

[0104] This comparative example is basically the same as Example 1, except that the cinnamon twig in step S1 is replaced with an equal amount of asarum.

[0105] Comparative Example 5

[0106] This comparative example is basically the same as Example 1, except that the asarum in step S1 is replaced with an equal amount of cassia twig.

[0107] Test example

[0108] Experiment 1:

[0109] After SD rats were killed by cervical dislocation, their abdominal hair was removed, and their abdominal skin was separated to ensure that the stratum corneum of the skin was intact, and the excess subcutaneous fat tissue and connective tissue were removed, and the skin was rinsed with physiological saline for later use. Then the mouse skin was randomly divided into 4 groups, namely, the microneedle group of the traditional Chinese medicine composition prepared in Examples 1-3 and the gel ointment prepared in Comparative Example 1, with 3 parts in each group.

[0110] Treatment method of the microneedle group of the Chinese medicine composition: take out the spare mouse skin, gently wipe off the excess moisture on the surface with clean filter paper, spread it flat on a clean foam board with the stratum corneum facing up, press the microneedles with a force of 20N and retain them on the skin, and stick the microneedles to the skin with medical tape to prevent them from falling off.

[0111] Treatment method for the gel ointment group: wipe off excess moisture from the spare mouse skin, spread it on the diffusion tank with the stratum corneum layer facing upward, and apply ordinary gel ointment on the skin.

[0112] Then the mouse skin was directly fixed between the diffusion cell and the receiving tank of the drug transdermal diffusion tester, with the keratin layer of the skin facing the diffusion cell and the dermis layer facing the receiving tank. 20 mL of 37°C physiological saline was added to the receiving tank as the receiving medium, and the bubbles were exhausted so that the receiving liquid surface was in full contact with the skin. The magnetic stirring was carried out at a speed of 300 r / min, and the 37°C constant temperature water bath was circulated. The timing was started after the administration, and 1 mL of the receiving solution was taken out at 0.5, 1, 2, 4, 6, 8, 10, 12, 24 and 48 hours, and the blank receiving medium of equal volume was added at the same temperature. The obtained receiving solution was passed through a microporous filter membrane with a pore size of 0.22 μm, and was measured by high performance liquid chromatography to calculate the cumulative permeability. The results are shown in Table 1.

[0113] Table 1: Comparison of cumulative permeability of microneedles and gel ointments of traditional Chinese medicine compositions

[0114] Group Example 1 Example 2 Example 3 Comparative Example 1 Cumulative permeability (%) 94.2 93.8 93.5 18.7

[0115] As can be seen from Table 1, the cumulative permeability of the microneedles of the traditional Chinese medicine compositions prepared in Examples 1-3 is higher than 93%, while the cumulative permeability of the gel paste prepared in Comparative Example 1 is only 18.7%, which is much lower than that of the microneedles of the traditional Chinese medicine compositions. It can be seen that by mixing degradable and highly biocompatible hyaluronic acid and chitosan to form a composite matrix solution, and then using the composite matrix solution and the mixed traditional Chinese medicine extract as raw materials to prepare the microneedles of the traditional Chinese medicine composition, since the microneedles of the traditional Chinese medicine composition are composed of swellable polymer cross-linked hydrogel, the needle tip pierces the skin, leaving a microporous channel, so that the mixed traditional Chinese medicine extract loaded by it absorbs interstitial fluid through the microporous channel with the carrier material, thereby expanding, so that the drug is continuously released from the carrier material to the site of action, thereby achieving the effect of improving the utilization efficiency of the drug.

[0116] Experiment 2:

[0117] The bacteria S. aureus, Escherichia coli, and MRSA were revived and inoculated onto solid culture medium and cultured in a 37°C incubator for 18 h. One colony was selected and added to 4 mL of liquid culture medium and incubated in a 37°C shaker for 4 h. The bacterial count was then assessed using UV-visible spectrophotometry (600 nm). Each bacterium was then diluted to 10 using sterile culture medium. 6CFU / mL, the bacterial solution was randomly divided into 5 groups, namely, Example 1-3 groups and Comparative Example 2-3 groups, each group had 3 portions (respectively, bacterial solution of S. aureus, Escherichia coli, and MRSA), each portion was 200 μL, and the microneedles of the Chinese medicine composition prepared in Example 1-3 were added to the Example 1-3 groups, and the microneedles of the Chinese medicine composition prepared in Comparative Example 2-3 were added to the Comparative Example 2-3 groups, and then incubated in a 37°C constant temperature incubator for 4 hours, and then 800 μL of sterile liquid culture medium was added to continue incubation overnight, and then each group was diluted to 10 with sterile liquid culture medium 2 CFU / mL, inoculate 100 μL onto the surface of the solid culture medium using the dilution spreading plate method, place it in a 37°C incubator and culture it for 18 h, record the number of colonies on each culture medium, and calculate the antibacterial rate. The results are shown in Table 2.

[0118] Table 2: Comparison of antibacterial rates between Example 1-3 and Comparative Example 2-3

[0119] Group Antibacterial rate (%) Example 1 Group 98.7 Example 2 Group 99.2 Example 3 Group 98.6 Comparative Example 2 35.3 Comparative Example 3 45.5

[0120] As can be seen from Table 2, the antibacterial rates of Examples 1-3 are all higher than 98%, while the antibacterial rates of Comparative Examples 2-3 are approximately 35.3% and 45.5%, respectively, which are much lower than those of Examples 1-3. It can be seen that by dissolving chitosan in acetic acid solution and then stirring and mixing with an aqueous solution of tannic acid, the amino groups of chitosan and the hydroxyl groups of tannic acid will produce hydrogen bonds, so that the tannic acid is fixed in the chitosan scaffold to form a gel network, and then the silver nitrate aqueous solution is added to reduce the anions by using the phenolic hydroxyl groups of the tannic acid molecules to generate nanosilver with antibacterial effects. Since the amino groups of chitosan are positively charged, they can adsorb negatively charged bacteria. Therefore, when chitosan is compounded with nanosilver, the antibacterial effect can be further improved.

[0121] Experiment 3 (clinical trial):

[0122] 180 volunteers with neck, shoulder, waist and leg pain were randomly divided into three groups, namely Example 1 group, Comparative Example 4 group and Comparative Example 5 group, with 60 people in each group.

[0123] Drug options:

[0124] The Example 1 group was treated with the microneedles of the Chinese medicine composition prepared in Example 1, the Comparative Example 4 group was treated with the microneedles of the Chinese medicine composition prepared in Comparative Example 4, and the Comparative Example 5 group was treated with the microneedles of the Chinese medicine composition prepared in Comparative Example 5. The microneedles were replaced every 2 days for 21 consecutive days.

[0125] Efficacy criteria:

[0126] Clinical recovery: TCM clinical symptoms and signs disappear or basically disappear;

[0127] Markedly effective: clinical symptoms and signs of TCM are significantly changed;

[0128] Effective: TCM clinical symptoms and signs have improved;

[0129] Ineffective: There was no significant improvement in clinical symptoms and signs of TCM.

[0130] The treatment results are shown in Table 3.

[0131] Table 3: Comparison of treatment results between Example 1 and Comparative Examples 4-5

[0132] Group Number of people treated Number of recovered people Number of people with significant effect Effective number of people Invalid number of people Total effective rate (%) Example 1 Group 60 50 8 2 0 100 Comparative Example 4 60 39 12 9 0 100 Comparative Example 5 60 42 10 8 0 100

[0133] As can be seen from Table 3, although the total effective rates of Example 1, Comparative Example 4 and Comparative Example 5 are all 100%, the number of cured people in Comparative Example 4 and Comparative Example 5 is less than that in Example 1. It can be seen that by first mixing and grinding Herba Mori and Radix Gentianae, mixing with distilled water, and adding cellulase for enzymolysis to prepare an enzymolysis solution, and then stirring and mixing the enzymolysis solution and ethanol solution with the mixed powder of Rhizoma Chuanxiong, Rhizoma Angelicae Pubescentis, Cinnamomum cassiae, Clematidis, Carthamus tinctorius, Achyranthes bidentata, Caulis Spatholobi and Asarum after grinding, and performing microwave-assisted heating reflux extraction, a mixed Chinese medicine extract with the effects of promoting blood circulation and removing blood stasis, dispelling wind and unblocking collaterals, anti-inflammatory and analgesic is prepared, which can effectively treat neck, shoulder, waist and leg pain, wherein Cinnamon twig and Asarum have a synergistic effect, and the simultaneous addition of the two can further improve the efficacy of the mixed Chinese medicine composition.

[0134] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A method for preparing a composition for treating neck, shoulder, waist and leg pain, characterized in that: The steps include: S1: Preparation of mixed Chinese herbal medicine extracts Mixing and grinding the herb and gentiana macrophylla, and fully mixing them with distilled water, then adding cellulase for enzymolysis to prepare an enzymolysis solution, and then mixing and grinding chuanxiong, angelica dahurica, cassia twig, clematis root, safflower, achyranthes bidentata, millettia spatholobi and asarum, and mixing them with the ethanol solution and the enzymolysis solution, and performing microwave-assisted heating reflux extraction to obtain a mixed Chinese medicine extract; The raw material components of the mixed Chinese medicine extract are as follows: 20-30 parts of Chuanxiong, 12-18 parts of Angelica dahurica, 12-16 parts of Cinnamomum cassia, 8-14 parts of Morus alba, 6-10 parts of Clematis, 7-9 parts of Carthamus tinctorius, 3-5 parts of Achyranthes bidentata, 3-5 parts of Millettia reticulata, 3-5 parts of Asarum and 1-3 parts of Gentiana macrophylla; S2: Preparation of nanosilver composite solution Dissolving chitosan and tannic acid in acetic acid solution and distilled water respectively to prepare chitosan-acetic acid solution and tannic acid solution, then stirring and mixing the tannic acid solution and the chitosan-acetic acid solution, adding silver nitrate aqueous solution, stirring and reacting to obtain a nano-silver composite solution; S3: Preparation of composite matrix solution Add hyaluronic acid into distilled water at a solid-liquid ratio of 1 g: (20-30) mL, stir until completely dissolved, then add the above-mentioned nanosilver composite solution, stir and mix evenly to obtain a composite matrix solution; S4: Preparation of microneedles of Chinese medicine composition The above-mentioned composite matrix solution is poured into a microneedle mold, and after centrifugation, freezing and thawing, a microneedle needle head mold is made. Then, the above-mentioned mixed Chinese medicine extract is added to the microneedle needle head mold, flattened and the composite matrix solution is added again. After centrifugation, freezing and thawing, it is dried and demolded to obtain a Chinese medicine composition microneedle.

2. The method for preparing a composition for treating neck, shoulder, waist and leg pain according to claim 1, characterized in that: S1 specifically includes the following steps: S1.1: Mix and grind the herb and Radix Gentianae Macrophyllae, add them into distilled water at a solid-liquid ratio of 1 g: (30-40) mL, heat and stir at 40-50 °C for 1-2 h to obtain a binary mixed suspension; S1.2: Add 0.3-0.5 wt% cellulase to the above binary mixed suspension, keep warm for 40-50 min, then heat to 90-100°C to inactivate the enzyme for 3-5 min to obtain an enzymatic solution; S1.3: Mix and grind Chuanxiong, Angelica dahurica, Cinnamomum cassia, Clematis chinensis, Carthamus tinctorius, Achyranthes bidentata, Millettia spatholobi and Asarum, add them into 60% ethanol solution at a solid-liquid ratio of 1 g: (40-50) mL, then add the above enzymatic hydrolysate, stir and mix evenly to obtain a composite suspension; S1.4: placing the composite suspension in a microwave extractor, performing microwave-assisted heating reflux extraction for 2 to 3 hours at a temperature of 80 to 90°C and a power of 80 to 100 W, and filtering to obtain an extract; S1.5: Place the above extract in a concentrator, heat and concentrate it, remove the solvent, and obtain a mixed Chinese medicine extract.

3. The method for preparing a composition for treating neck, shoulder, waist and leg pain according to claim 2, characterized in that: S2 specifically includes the following steps: S2.1: Add chitosan to acetic acid solution at a solid-liquid ratio of 1 g: (20-30) mL, stir thoroughly to dissolve, and prepare chitosan-acetic acid solution; S2.2: Add tannic acid to distilled water, heat and stir at 70-90°C until fully dissolved, to obtain a 200-300 μg / mL tannic acid solution; S2.3: Add the above tannic acid solution to the above chitosan-acetic acid solution in a volume ratio of 1: (3-4), stir and mix evenly, then add the silver nitrate aqueous solution, continue stirring for 2-3 hours, react and obtain a nanosilver composite solution.

4. The method for preparing a composition for treating neck, shoulder, waist and leg pain according to claim 3, characterized in that: S4 specifically includes the following steps: S4.1: Pour the composite matrix solution prepared in step S3 into the microneedle mold, place it in a centrifuge, centrifuge it horizontally at a rate of 3000-4000 r / min for 4-6 minutes, then rotate it 180 degrees, continue centrifugation for 4-6 minutes, repeat 2-3 times, take it out, place it in a freezer, freeze it at -50--40°C for 1-2 hours, and finally thaw it in a medium-heat microwave for 20-30 seconds to obtain a microneedle needle mold; S4.2: Add the mixed Chinese medicine extract obtained in step S1.5 to the above-mentioned microneedle needle mold, flatten it, add the composite matrix solution, centrifuge at a rate of 3000-4000 r / min for 4-6 minutes, then rotate 180 degrees, continue centrifugation for 4-6 minutes, repeat 3-5 times, and obtain a microneedle backing mold; S4.3: Place the above-mentioned microneedle backing mold in a freezer and freeze it at -50 to -40°C for 1 to 2 hours. After taking it out, thaw it in a microwave at medium heat for 20 to 30 seconds. Repeat the freeze-thaw process 3 to 4 times, dry it at room temperature for 20 to 24 hours and demold it to obtain the traditional Chinese medicine composition microneedles.

5. The method for preparing a composition for treating neck, shoulder, waist and leg pain according to claim 3, characterized in that: The volume fraction of the acetic acid solution is 2 to 4%.

6. The method for preparing a composition for treating neck, shoulder, waist and leg pain according to claim 3, characterized in that: The volume ratio of the silver nitrate aqueous solution to the tannic acid solution is 1:(2-3), and the concentration of the silver nitrate aqueous solution is 10-20 μg / mL.

7. The method for preparing a composition for treating neck, shoulder, waist and leg pain according to claim 1, characterized in that: The mass ratio of hyaluronic acid to chitosan in the nanosilver composite solution is (3-5):

1.

8. The method for preparing a composition for treating neck, shoulder, waist and leg pain according to claim 4, characterized in that: The mass ratio of the mixed Chinese herbal medicine extract to the composite matrix solution is 1:(400-500).

9. Use of a composition prepared by the method for preparing a composition for treating cervical, shoulder, waist and leg pain according to any one of claims 1 to 8 in preparing a medicament for treating cervical, shoulder, waist and leg pain.