External anti-inflammatory traditional Chinese medicine composition and its application, anti-inflammatory gel patch and its preparation method

Abstract

This invention discloses a topical anti-inflammatory traditional Chinese medicine composition and its application, as well as an anti-inflammatory gel patch and its preparation method, belonging to the field of traditional Chinese medicine technology. The topical anti-inflammatory traditional Chinese medicine composition, by weight, comprises the following components: 30-120 parts of *Sinomenium acutum* and 0.35-1.4 parts of *Ilex cornuta* oil. The combination of the active components *Sinomenium acutum* and *Ilex cornuta* oil achieves a synergistic effect, ensuring the efficacy of the active ingredients. Pharmacological studies have shown that the topical anti-inflammatory traditional Chinese medicine composition of this invention exhibits significant therapeutic effects in alleviating biomarkers related to osteoarthritis, frozen shoulder, soft tissue injury, and muscle pain, which is of great significance for improving patients' quality of life.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and particularly relates to an external anti-inflammatory traditional Chinese medicine composition and its application, an anti-inflammatory gel plaster and its preparation method. Background Art

[0002] Sinomenium acutum has been used as a traditional Chinese medicine for treating rheumatism and arthralgia in China for more than 1000 years. It has obvious effects when used alone and is often combined with Stephania tetrandra. It is decocted with wine for drinking (in "Prescriptions for Universal Relief"); or used in combination with Sargentodoxa cuneata, Saposhnikovia divaricata, Cinnamomum cassia, etc.; for shoulder and arm pain, it can be combined with Curcuma longa and Notopterygium incisum; for waist and knee pain, it can be combined with Angelica pubescens and Achyranthes bidentata. "Compendium of Materia Medica": "Treat rheumatism and arthralgia, pain in joints and knees, paralysis, itching, injuries and sores. Used in medicinal wine." "Synopsis of Materia Medica": "Sinomenium acutum is a medicine for dispersing wind-cold-damp arthralgia. It can soothe muscles and activate blood circulation, reduce bone fracture and benefit marrow. Therefore, for the syndrome of weakness and paralysis caused by wind diseases, long-term use can achieve great effects."

[0003] Modern research has found that the main components of Sinomenium acutum are sinomenine, sinomenine, michelalbine, stepholidine, magnoflorine, tetrahydroepiberberine, isosinomenine, sutchuenine and other alkaloids. The isoquinoline alkaloid monomer extracted from Sinomenium acutum, namely sinomenine, has pharmacological effects such as analgesia, sedation and local anesthesia, and can also reduce hypertension, anti-inflammatory, anti-rheumatic, rheumatoid arthritis, etc. The clinically used sinomenine preparation mainly uses its hydrochloride, namely sinomenine hydrochloride, for treating rheumatoid arthritis, and its therapeutic effect is relatively significant. The currently marketed sinomenine hydrochloride preparations are mainly Zhengqingfengtongning tablets, Zhengqingfengtongning capsules, Zhengqingfengtongning sustained-release tablets and Zhengqingfengtongning injections. When administered orally, due to the short biological half-life of sinomenine hydrochloride, the oral dose is very large. For Zhengqingfengtongning tablets, 12 tablets need to be taken daily, for Zhengqingfengtongning capsules, 9 capsules need to be taken daily, and for Zhengqingfengtongning sustained-release tablets, 4 tablets need to be taken daily; when administered by injection, due to the instability of sinomenine to light and heat, it is easy to decompose, resulting in a decrease in efficacy and difficulty in preservation. In addition, when sinomenine hydrochloride preparations are used to treat rheumatoid arthritis, the onset time of its analgesic effect is relatively slow. Research shows that patients generally start to take effect 2 weeks after taking the medicine and significantly improve after 1 month. At the same time, sinomenine itself is a strong histamine releaser, and a large amount of histamine will be released after oral administration, causing gastrointestinal adverse reactions such as stomachache, nausea, diarrhea, etc. In severe cases, gastric ulcer and bleeding may occur; skin adverse reactions such as skin flushing, itching, burning, rash, etc.; other adverse reactions such as dizziness, headache, sweating, etc. A few patients will also experience leukopenia and thrombocytopenia. When administered by injection, many patients will experience itching, flushing, sweating, and aggravated swelling and pain 1-10 minutes after injection, and it takes 0.5-1 hour to relieve. If the reaction is very severe, dexamethasone or promethazine needs to be injected for counteraction; in addition, there are also case reports of anaphylactic shock.

[0004] Wintergreen oil, also known as wintergreen oil, is primarily composed of methyl salicylate. Wintergreen oil has a wide range of sources; it can be extracted from various plants such as *Ilex yunnanensis*, *Santalum album*, and *Ilex cornuta*. Methyl salicylate is a nonsteroidal anti-inflammatory drug (NSAID). It primarily exerts its anti-inflammatory and analgesic activity by inhibiting cyclooxygenase, thereby reducing prostaglandin synthesis. It can relieve local swelling and pain caused by sprains, contusions, strains, overuse injuries, muscle and fasciitis, traumatic synovitis, and ligament injuries.

[0005] Traditional Chinese medicine (TCM) external dosage forms suffer from rudimentary preparation processes, inadequate matrix selection, and insufficient understanding of transdermal absorption mechanisms. Furthermore, TCM compound preparations, due to their complex composition (often involving multiple ingredients working together for therapeutic effect) and generally requiring larger dosages, have historically experienced slow development in novel external TCM dosage forms. A TCM compound composed of *Sinomenium acutum* (Qingfengteng) and *Ilex chinensis* oil, when developed into external dosage forms using modern pharmaceutical technology, such as gels, gel patches, ointments, liniments, and films, is suitable for wind-cold-dampness syndrome, characterized by muscle soreness, joint swelling, pain, limited flexion and extension, numbness and stiffness, as well as rheumatoid arthritis patients exhibiting the aforementioned symptoms. Given that these diseases primarily manifest in local joints, developing a transdermal topical formulation of *Sinomenium acutum*, *Ilex chinensis* oil, and menthol is highly appropriate.

[0006] Currently, there are no studies or reports on the preparation of topical dosage forms using Sinomenium acutum and wintergreen oil as the main medicinal ingredients. Summary of the Invention

[0007] To address the aforementioned technical problems, this invention provides a topical anti-inflammatory traditional Chinese medicine composition, its preparation method, and its application. The topical anti-inflammatory traditional Chinese medicine composition of this invention utilizes the synergistic effect of Sinomenium acutum and Ilex cornuta oil to enhance the anti-inflammatory effect and ensure the efficacy of the active ingredients.

[0008] To achieve the above objectives, the present invention adopts the following technical solution:

[0009] A topical anti-inflammatory traditional Chinese medicine composition comprising the following components: Chinese pine and wintergreen oil.

[0010] In this invention, *Sinomenium acutum* is the principal ingredient, bitter, pungent, and neutral in nature, entering the liver and spleen meridians. It effectively dispels wind and dampness, and unblocks the meridians, especially commonly used for rheumatic pain and joint swelling. Wintergreen oil is primarily an extract of *Phyllostachys yunnanensis*, which is pungent and warm, dispelling wind and dampness, promoting qi and blood circulation, unblocking the meridians and relieving pain, and is used to treat rheumatic pain; it is the assistant ingredient. Menthol is an extract of peppermint. Peppermint is pungent, and its pungent nature allows it to disperse. For external use, its aroma and ability to open the orifices are utilized, allowing it to effectively penetrate the skin surface and guide other external medicines into the nutritive and defensive qi to relieve stagnation; it is the guiding ingredient. The combined effects of these ingredients dispel wind and dampness, promote blood circulation, unblock the meridians, promote qi circulation, and relieve pain.

[0011] In some preferred embodiments, the amount of *Sinomenium acutum* in the topical anti-inflammatory traditional Chinese medicine composition of the present invention is 30 to 120 parts by weight, preferably 30, 60, 90, or 120 parts or any value between the above. More preferably, it is 60 to 90 parts.

[0012] In some preferred embodiments, the amount of wintergreen oil in the topical anti-inflammatory traditional Chinese medicine composition of the present invention is 0.35 to 1.4 parts by weight, preferably 0.35 parts, 0.7 parts, 1.05 parts, 1.4 parts, or any value between the above. More preferably, it is 0.7 to 1.05 parts.

[0013] In some other preferred embodiments, the topical anti-inflammatory traditional Chinese medicine composition comprises, by weight, the following components: 30-120 parts of Sinomenium acutum and 0.35-1.4 parts of wintergreen oil.

[0014] In some other preferred embodiments, the topical anti-inflammatory traditional Chinese medicine composition comprises, by weight, the following components: 60-90 parts of Sinomenium acutum and 0.7-1.05 parts of wintergreen oil.

[0015] In some preferred embodiments, the topical anti-inflammatory traditional Chinese medicine composition of the present invention, by weight, comprises the following components: Chinese sedum, wintergreen oil and menthol.

[0016] In some preferred embodiments, the amount of menthol in the topical anti-inflammatory traditional Chinese medicine composition of the present invention, by weight percentage, is 0.3 to 1.2 parts, preferably 0.3 parts, 0.6 parts, 0.9 parts, 1.2 parts or any value between the above values.

[0017] In some other preferred embodiments, the topical anti-inflammatory traditional Chinese medicine composition comprises, by weight, the following components: 30-120 parts of Sinomenium acutum, 0.35-1.4 parts of wintergreen oil, and 0.3-1.2 parts of menthol.

[0018] In some other preferred embodiments, the topical anti-inflammatory traditional Chinese medicine composition comprises, by weight, the following components: 60-90 parts of Sinomenium acutum, 0.7-1.05 parts of wintergreen oil, and 0.6-1.2 parts of menthol.

[0019] In a more preferred embodiment, the topical anti-inflammatory traditional Chinese medicine composition comprises, by weight, the following components: 120 parts of Sinomenium acutum, 1.4 parts of wintergreen oil, and 1.2 parts of menthol.

[0020] In some other preferred embodiments, the wintergreen oil includes synthetic sources and extraction sources; wherein the synthetic sources include synthetic methyl salicylate; and the extraction sources include extracts obtained from plants, the extracts including any one or more of the following: *Gynostemma pentaphyllum* extract, *Gynostemma pentaphyllum* extract, *Gynostemma pentaphyllum* extract, *Santalum album* extract, wintergreen extract, and birch extract.

[0021] In some other preferred embodiments, the wintergreen oil contains more than 90% methyl salicylate.

[0022] In some other preferred embodiments, the topical anti-inflammatory traditional Chinese medicine composition contains *Sinomenium acutum* extracted with an aqueous ethanol solution, concentrated, and dried to obtain *Sinomenium acutum* extract.

[0023] Preferably, the concentration of the ethanol aqueous solution is 20-80%.

[0024] The present invention also provides the application of the above-mentioned topical anti-inflammatory traditional Chinese medicine composition in the preparation of products for treating osteoarthritis, frozen shoulder, soft tissue injury and muscle pain.

[0025] Preferably, the dosage form of the product for treating osteoarthritis, frozen shoulder, soft tissue injury, and muscle pain is a gel, gel patch, ointment, liniment, or film-forming agent. Those skilled in the art will understand that the dosage form is prepared by adding some functional excipients to the above-mentioned topical anti-inflammatory traditional Chinese medicine composition to the corresponding dosage form formulation.

[0026] More preferably, the dosage form of the product for treating osteoarthritis, frozen shoulder, soft tissue injury, and muscle pain is a gel patch.

[0027] The present invention also provides an anti-inflammatory gel patch comprising the above-mentioned topical anti-inflammatory traditional Chinese medicine composition.

[0028] Preferably, the anti-inflammatory gel patch further includes a gel matrix, which comprises a polymer backbone material, a thickener, a moisturizer, a pH adjuster, a crosslinking agent, a crosslinking regulator, a surfactant, a filler, and water.

[0029] The polymeric framework material used in this invention is a bioadhesive material with low skin irritation commonly used in the art, including one or more of sodium polyacrylate, partially neutralized sodium polyacrylate, carbomer, and polyvinyl alcohol. A combination of partially neutralized sodium polyacrylate and polyvinyl alcohol is preferred. The tackifier used in this invention is one or more of gelatin, sodium carboxymethyl cellulose, methyl acrylate-2-ethylhexyl acrylate copolymer resin emulsion, polyacrylic acid solution, povidone solution, acrylic acid grafted starch, and polyvinylpyrrolidone. Preferably, the tackifier is one or more of gelatin, sodium carboxymethyl cellulose, polyacrylic acid solution, and methyl acrylate-2-ethylhexyl acrylate copolymer resin emulsion. The humectant used in this invention is one or more of glycerin, propylene glycol, 1,3-butanediol, urea, sorbitol, and polyethylene glycol, preferably glycerin. The pH adjuster used in this invention is tartaric acid, lactic acid, malic acid, or L-tartaric acid. The present invention uses one or more of the following: citric acid, phosphoric acid, and preferably tartaric acid; the crosslinking agent used in the present invention is one or more of the following: aluminum hydroxide, aluminum hydroxide, aluminum oxide, aluminum hydroxide gel, and aluminum trichloride, preferably aluminum hydroxide and aluminum hydroxide; the crosslinking regulator used in the present invention is one or more of the following: disodium edetate and tetrasodium edetate, preferably disodium edetate; the surfactant used in the present invention is one or more of the following: Span 20, Span 40, Span 60, polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80, preferably polysorbate 80; the filler used in the present invention is one or more of the following: kaolin, talc, titanium dioxide, micronized silica, silica, and diatomaceous earth, preferably kaolin.

[0030] The present invention also provides a method for preparing the above-mentioned anti-inflammatory gel patch, comprising the following steps:

[0031] (1) Preparation of Sinomenium acutum extract: Sinomenium acutum was pulverized, extracted with ethanol aqueous solution, concentrated and dried to obtain Sinomenium acutum extract;

[0032] (2) Preparation of aqueous mixture: Add crosslinking regulator, humectant, emulsifier and part of thickener to water, then add Sinomenium acutum extract, mix well to obtain aqueous mixture;

[0033] (3) Preparation of oil phase mixture: Add the skeleton material, crosslinking agent and filler to glycerol, mix well to obtain oil phase mixture;

[0034] (4) Preparation of the main drug mixture: Mix the surfactant, wintergreen oil and menthol evenly to obtain the main drug mixture;

[0035] (5) Prepare pH adjustment solution: Dissolve the pH adjustment agent in water;

[0036] (6) Mixing and preparing the ointment: Mix the oil phase mixture, the aqueous phase mixture and the main drug mixture evenly, then add the pH adjuster solution and the thickener, mix evenly to obtain the total ointment;

[0037] (7) Coating and cutting: Coat and cut the total mixture evenly to obtain the gel patch.

[0038] Preferably, no acid or alkali is added during the extraction process in step (1).

[0039] Preferably, the concentration of the ethanol aqueous solution in step (1) is 20-80%.

[0040] This invention also relates to the use of the gel patch in the preparation of drugs for treating osteoarthritis, frozen shoulder, soft tissue injury, and muscle pain.

[0041] This invention also relates to the use of wintergreen oil in the preparation of anti-inflammatory gel patches. Wintergreen oil can simultaneously act as an active ingredient and a penetration enhancer in anti-inflammatory gel patches, and is therefore used as both an active ingredient and a penetration enhancer. In some embodiments, this invention relates to the use of wintergreen oil as both an active ingredient and a penetration enhancer in the preparation of anti-inflammatory gel patches.

[0042] By adopting the above technical solution, the beneficial effects of the present invention are as follows:

[0043] (1) The main ingredients of the topical anti-inflammatory Chinese medicine composition of the present invention are Sinomenium acutum and wintergreen oil (which also acts as a penetration enhancer), with optional addition of menthol (which also acts as a penetration enhancer). Wintergreen oil, by utilizing its own lipid-soluble properties, to a certain extent destroys the lipid structure of the stratum corneum, changes the dense arrangement of the surface stratum corneum, reduces the skin barrier function, and thus promotes the absorption of the drug.

[0044] (2) The anti-inflammatory gel patch of the present invention has high permeability by adding wintergreen oil. According to the long-term stability study of 12 months and the in vitro release test, the anti-inflammatory gel patch of the present invention has stable in vitro release and stable quality, and meets the pharmaceutical standards.

[0045] (3) The product prepared using the anti-inflammatory Chinese herbal extract of the present invention has comparable efficacy to similar products already on the market (Zhengqing Fengtongning Injection), and has fewer side effects and is safe and reliable.

[0046] Terminology Definition

[0047] In this invention, the term "Qingfengteng extract" refers to the Qingfengteng extract obtained by pulverizing the dried stems of the Menispermaceae plant Qingfengteng or Maoqingteng, extracting them with an ethanol solution (including but not limited to one or more of reflux extraction, soaking extraction or percolation extraction), concentrating the extract, and drying it.

[0048] In this invention, the term "wintergreen oil," also known as wintergreen oil, mainly consists of methyl salicylate, with a content of 90% or more. Preferably, it is 90% to 100%. In some embodiments, the content of methyl salicylate in wintergreen oil is 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, or any value within the range of any two of these values.

[0049] Wintergreen oil has a wide range of sources, mainly divided into synthetic and extracted sources. Synthetic sources include, but are not limited to, synthetic methyl salicylate, such as commercially available wintergreen oil (also known as wintergreen oil) or commercially available synthetic methyl salicylate; or the preparation method of said wintergreen oil includes chopping plants, such as *Gynura divaricata*, *Gynura pseudochina*, *Santalum album*, *Gynura pseudochina*, and / or wintergreen, extracting by steam distillation, collecting the lower volatile oil to obtain said wintergreen oil. Extracted sources include, but are not limited to: extracting extracts from plants to obtain extracts, said extracts including any one or more of *Gynura divaricata* extract, *Gynura pseudochina* extract, *Santalum album* extract, wintergreen extract, and birch extract. Attached Figure Description

[0050] Figure 1 The cumulative release curve of sinomenine in the anti-inflammatory gel patch over 12 hours.

[0051] Figure 2 This is the HPLC chromatogram of the extract of *Sinomenium acutum*.

[0052] Figure 3 This is the GC chromatogram of wintergreen oil. Detailed Implementation

[0053] The present application will be further described in detail below through specific embodiments. The following embodiments are only for further illustration of the present application and should not be construed as limiting the present application.

[0054] The reagents and raw materials used in this invention are all commercially available.

[0055] Examples 1-4 and Comparative Examples 1-5

[0056] The formulations of the gels in Examples 1-4 and Comparative Examples 1-5 are shown in Table 1.

[0057] Table 1

[0058]

[0059] Manufacturing method

[0060] (1) Preparation of Sinomenium acutum extract:

[0061] S1 Extraction: Crush *Sinomenium acutum*, moisten with 40% ethanol for 2 hours, then add an appropriate amount of 40% ethanol and soak for 24 hours. Percolate to obtain percolate for later use.

[0062] S2 Concentration: The percolate is transferred to a concentrator and concentrated under reduced pressure to 1.0~1.3 g / cm³. 3 .

[0063] S3 ointment preparation: The concentrated liquid is dried under reduced pressure until the moisture content is below 10% to obtain the extract of Sinomenium acutum.

[0064] (2) Preparation of the main drug mixture: Add wintergreen oil, menthol, 4 parts of polysorbate 80 to 40 parts of glycerin and mix well.

[0065] (3) Preparation of gel: Add 3 parts of sodium carboxymethyl cellulose to 50 parts of water and let it swell; then add the extract of Sinomenium acutum, mix evenly, then add the main drug mixture and mix evenly to obtain the gel.

[0066] Examples 5-8 and Comparative Examples 6-10

[0067] The formulations of the anti-inflammatory gel patches in Examples 5-8 and Comparative Examples 6-10 are shown in Table 2.

[0068] Table 2

[0069]

[0070] Manufacturing method

[0071] (1) Preparation of Sinomenium acutum extract: Same as in Examples 1-4.

[0072] (2) Preparation of aqueous mixture: Add gelatin, disodium edetate, sodium carboxymethyl cellulose and polyvinyl alcohol to water, then add sinomenium extract, mix well to obtain aqueous mixture.

[0073] (3) Preparation of oil phase mixture: Add part of the neutralized sodium polyacrylate, aluminum hydroxyl and kaolin to part of the glycerol, mix well to obtain oil phase mixture.

[0074] (4) Preparation of the main drug mixture: Add polysorbate 80, wintergreen oil and menthol to part of the glycerin and stir well.

[0075] (5) Prepare tartaric acid solution: Dissolve tartaric acid in water.

[0076] (6) Mixing and preparing the ointment: Mix the oil phase mixture, the water phase mixture and the main drug mixture evenly, then add tartaric acid solution and polyacrylic acid solution, mix evenly to obtain the total ointment.

[0077] (7) Coating and cutting: Coat and cut the total mixture evenly to obtain the gel patch.

[0078] Application examples

[0079] A rat osteoarthritis model was constructed using biochemical methods to investigate the therapeutic effect of an anti-inflammatory gel on the rat osteoarthritis model. A positive control group was set up as a control to evaluate and compare the effects of the anti-inflammatory gel on the pain threshold and the swelling circumference of the joint in rat osteoarthritis, and to explore the pharmacodynamic effects of different administration methods and dosages on osteoarthritis.

[0080] Effects of anti-inflammatory gel on a rat model of osteoarthritis

[0081] 132 qualified SPF-grade male SD rats were selected and divided into a normal group (n=12) and a model group (n=120). The model group rats received a single injection of 0.1 mL of 4% papain solution per rat into their right posterior knee joint cavity on days 1, 4, and 7. The normal group rats received an equal volume of 0.9% sodium chloride injection, for a total of 3 injections. After modeling, the rats continued feeding for one week, and then two animals from each group were randomly selected for euthanasia to collect cartilage tissue for histopathological examination. 110 rats with significant swelling at the modeling site were randomly divided into 11 groups according to body weight: the model control group, the Zhengqing Fengtongning injection group, Examples 1-4, and Comparative Examples 1-5, with 10 animals in each group. The injection group received Zhengqing Fengtongning injection intramuscularly at 5 mg / kg twice daily. The other groups received the corresponding test substance directly applied to the front of the right knee joint of the rats, covering an area of ​​2.5 cm × 2.5 cm, and fixed with cellophane for 12 hours to prevent drug detachment. The normal control group and the model control group were given the same amount of blank matrix, and the right joint was fixed for 12 hours every day. The drugs were administered for 7 consecutive days. The swelling circumference of the rat joint was measured the day after the last administration. The pain threshold of the joint of each group of rats was measured using a pain pressure meter. Joint fluid was collected and biomarkers such as PGE, COX-2, IL-1β, and TNF-α were detected.

[0082] Experimental results

[0083] (1) Effect on bone and joint swelling in rats

[0084] As shown in Table 3, compared with the normal control group, the swelling circumference of the bone and joint in the model control group was significantly increased ( P ≤0.01); Compared with the model control group, except for the 5 control groups, the circumference of joint swelling in rats in the other treatment groups was significantly reduced ( P (≤ 0.01); compared with the control group 5, the swelling circumference of the joints of rats in the other treatment groups was significantly reduced; compared with the control groups 1-4, the swelling circumference of the joints of rats in the Example 1-4 groups was significantly reduced; compared with the Zhengqing Fengtongning injection group, there was no significant difference in the swelling circumference of the joints of rats in the Example 1-4 groups, and they were at the same level.

[0085] Table 3. Effects of anti-inflammatory gel on swelling perimeter in a rat model of osteoarthritis ( (n=10)

[0086]

[0087] Note: Compared with the normal control group, + P ≤ 0.01; compared with the model control group, ** P ≤ 0.01.

[0088] (2) Effect on pain threshold in rat osteoarthritis model

[0089] As shown in Table 4, compared with the normal control group, the bone and joint pain threshold of rats in the model control group was significantly reduced before drug administration and on the day after the last drug administration. P ≤ 0.01); Compared with the model control group, except for the control group 5, the bone and joint pain threshold of all other treatment groups was significantly increased on the day after the last administration (P<0.01); Compared with the control group 5, the bone and joint pain threshold of other treatment groups was significantly increased on the day after the last administration; Compared with the Zhengqing Fengtongning injection group, the bone and joint pain threshold of the last administration of the first and second examples was slightly lower, while there was no significant difference in the bone and joint pain threshold of the last administration of the third and fourth examples; Compared with the first and fourth examples, the bone and joint pain threshold of the control group was significantly lower on the day after the last administration.

[0090] Table 4. Effects of anti-inflammatory gel on the bone and joint pain threshold in rats ( (n=10)

[0091]

[0092] Note: Compared with the normal control group + P ≤0.01; compared with the model control group ** P ≤0.01.

[0093] (3) Inflammatory factors in the joint fluid

[0094] As shown in Table 5, compared with the normal control group, the levels of IL-1β and TNF-α in the model control group were significantly increased ( P ≤0.05); Compared with the model control group, there were no significant changes in IL-1β and TNF-α levels in the five comparative groups, while the levels of IL-1β and TNF-α in the Zhengqing Fengtongning injection group, Examples 1-4 groups, and Comparative Examples 1-4 groups were significantly reduced ( P ≤ 0.05 or P ≤ 0.01).

[0095] Table 5. Effects of anti-inflammatory gel on inflammatory factors in a rat model of osteoarthritis ( (n=10)

[0096]

[0097] Note: Compared with the normal control group, + P ≤ 0.05, ++ P ≤ 0.01; compared with the model control group, * P ≤0.05, ** P ≤ 0.01.

[0098] (4) Changes in PGE and COX-2 in the joint fluid

[0099] As shown in Table 6, compared with the normal control group, the levels of PGE and COX-2 in the model control group were significantly increased ( P ≤0.01); Compared with the model control group, there were no significant changes in PGE and COX-2 levels in the 5 comparative groups, while the PGE and COX-2 levels in the Zhengqing Fengtongning injection group, Examples 1-4 groups, and Comparative Examples 1-4 groups were significantly reduced ( P ≤ 0.05 or P (≤ 0.01); Compared with the Zhengqing Fengtongning injection group, the PGE and COX-2 levels in Example 1 group and Comparative Examples 1-4 were significantly increased, while the PGE and COX-2 levels in Example 2-4 were slightly decreased.

[0100] Table 6. Effects of anti-inflammatory gel on PGE and COX-2 in a rat model of osteoarthritis ( (n=10)

[0101]

[0102] Note: Compared with the normal control group, + P ≤ 0.01; compared with the model control group, * P ≤ 0.05, ** P ≤0.01.

[0103] This study induced an osteoarthritis model in rats using biochemical methods, while setting up a positive control drug group as a control, in order to evaluate and compare the efficacy of traditional Chinese medicine combinations on osteoarthritis.

[0104] The observation results are as follows

[0105] (1) Effect on bone and joint pain threshold: The effect of raising the pain threshold in the control group 5 was significantly lower than that in the other administration groups, indicating that the analgesic effect of single wintergreen oil was significantly weaker than that of single sinomenium and sinomenium wintergreen oil compound; the effect of raising the bone and joint pain threshold in rats in Examples 1-4 on the day after the last administration was significantly better than that in the control group 1-4, indicating that adding wintergreen oil helps to improve the analgesic effect of anti-inflammatory gel. Compared with Zhengqing Fengtongning injection, the effect of raising the bone and joint pain threshold in Example 1 was slightly worse, while the effect of raising the bone and joint pain threshold in Examples 2-4 was basically the same.

[0106] (2) Effect on the circumference of joint swelling: The effect of reducing the circumference of joint swelling in the control group 5 was significantly lower than that in the other treatment groups, indicating that the swelling-reducing effect of single wintergreen oil was significantly weaker than that of single sinomenium and sinomenium wintergreen oil compound; the effect of reducing the circumference of joint swelling in the rats in Examples 1-4 on the day after the last administration was significantly better than that in the control group 1-4, indicating that adding wintergreen oil helps to improve the swelling-reducing effect of anti-inflammatory gel. Compared with Zhengqing Fengtongning injection, the effect of reducing the circumference of joint swelling in Examples 1-4 was basically the same.

[0107] (3) Effects on osteoarthritis inflammatory factors in rats: There were no significant changes in joint inflammatory factors in the five comparative groups; the Zhengqing Fengtongning injection group, the groups of Examples 1-4, and the groups of Comparative Examples 1-4 could all effectively reduce the level of osteoarthritis inflammatory factors in rats, and there was no significant difference in the reduction effect.

[0108] (4) Effects on PGE and COX-2 in rat joint fluid: There were no significant changes in PGE and COX-2 in the joint fluid of the five comparative groups; the Zhengqing Fengtongning injection group, the groups of Examples 1-4 and the comparative groups 1-4 could effectively reduce the levels of PGE and COX-2 in the joint fluid of rats. The effects of the Zhengqing Fengtongning injection group and the groups of Examples 2-4 were basically the same, while the effects of the groups of Examples 1 and the comparative groups 1-4 were slightly worse.

[0109] Initial tack test of anti-inflammatory gel patch

[0110] Referring to Method 1 (Determination of Initial Tack) of General Chapter 0952, Part IV of the Chinese Pharmacopoeia (2020 Edition), the initial tack was determined using the rolling ball ramp stop method. Three test samples were taken, the protective layer removed, and placed in the center of an inclined plate with a 30° angle. The upper end of the test sample should be positioned below the horizontal line of the inclined plate, with the viscous surface facing upwards and the experimental length along the inclined plane not exceeding 5 cm. Steel balls of different diameters were placed at the horizontal line and rolled freely down the inclined plane. The initial tack was evaluated based on the number of the largest steel ball that the viscous surface could adhere to. The results showed that in Examples 5, 6, 7, 8 and Comparative Examples 6, 7, 8, 9, the Sinomenium acutum gel patches adhered to steel balls numbered 25, 26, 24, 26, 22, 22, 21, and 23, respectively, indicating good initial tack performance.

[0111] Excipient properties test of anti-inflammatory gel patch

[0112] Take one patch of each of the Qingfengteng gel plaster prepared in Examples 5-8 and Comparative Examples 6-9, place them in a constant temperature and humidity chamber at 37°C and 64% relative humidity for 30 minutes, take them out, and use clips to fix the test sample on a flat steel plate with the steel plate tilted at an angle of 60° to the horizontal plane. After 24 hours, there should be no dripping on the plaster surface.

[0113] The results showed that none of the batches of samples exhibited any flow, meeting the excipient testing requirements of the 2020 edition of the Chinese Pharmacopoeia.

[0114] Stability study of anti-inflammatory gel patch

[0115] According to the stability plan, the stability of four batches of self-made samples (samples from Examples 5, 6, 7, and 8) were investigated for 12 months. The results of the sample quality stability are shown in Table 7.

[0116] Table 7. Stability Study over 12 Months

[0117]

[0118] The above experimental results show that the anti-inflammatory gel patch of the present invention has excellent long-term stability, maintains stable quality after 12 months of storage, has a long shelf life, and meets pharmaceutical standards.

[0119] Investigation of in vitro release assay of anti-inflammatory gel patch

[0120] The anti-inflammatory gel patches of Examples 5-8 and Comparative Examples 6-9 were selected for in vitro release experiments. The experimental instrument was a TK-24BL transdermal ...

[0121] Experimental Procedure: Using a polyethersulfone membrane (0.45 μm pore size) as the separator and pH 7.4 PBS at 32 ± 0.5 °C as the receiving medium, the in vitro release characteristics of each sample through a vertical diffusion cell were determined. All receiving liquid was collected at 1, 2, 4, 6, 8, and 12 hours, with an equal volume of receiving liquid added simultaneously. The collected receiving liquid was filtered through a 0.45 μm filter membrane, and the filtrate was used as the test solution. The content of the main component in the solution was determined, and the result was calculated.

[0122] Figure 1 This is a schematic diagram illustrating the in vitro release results of the anti-inflammatory gel patch. Figure 1 It can be seen that the in vitro release rate of the anti-inflammatory gel patch in the embodiment is significantly better than that in the comparative example, indicating that the onset rate of the anti-inflammatory gel patch of the present invention is better than that of the single Sinomenium acutum extract gel patch.

[0123] Pharmacological study of anti-inflammatory gel patches

[0124] This invention uses Freund's complete adjuvant to induce a rat model of frozen shoulder, while setting up a positive control group as a control, to evaluate and compare the effects of anti-inflammatory gel patch application on the pain threshold of rat shoulder joint and the swelling circumference of rat shoulder joint, and to explore the pharmacological effects of different administration methods and dosages on osteoarthritis, frozen shoulder, soft tissue injury and muscle pain.

[0125] Effects of anti-inflammatory gel patch on a rat model of shoulder arthritis

[0126] 132 qualified SPF-grade male SD rats were selected and divided into a normal group (n=12) and a model group (n=120). In each group, the lateral aspect of the right shoulder of the rats was treated with hair removal on a 5cm × 5cm area. The rats in the model group received a single injection of 0.1 mL of Freund's complete adjuvant into the right anterior shoulder joint cavity on days 1, 5, 9, and 13. The normal control group received an equal volume of 0.9% sodium chloride injection, for a total of 4 injections. After modeling, two animals from each group were randomly selected and euthanized to collect the synovium of the shoulder joint and surrounding ligaments and tendons for histopathological examination. 110 rats with significant swelling at the modeling site were randomly divided into 11 groups according to body weight: the model control group, the Zhengqing Fengtongning injection group, the Examples 5-8 groups, and the Comparative Examples 6-10 groups, with 10 animals in each group. The injection group received 5 mg / kg of Zhengqing Fengtongning injection intramuscularly twice daily; the other groups applied the corresponding test substance directly to the right anterior shoulder joint, using medical breathable adhesive tape to fix the plaster to prevent it from falling off, twice daily, with the second dose removed before the next day's dose, and the two doses were 12 hours apart; the normal control group and the model control group received blank plasters of equal area applied using the same administration method, for 7 consecutive days.

[0127] The shoulder pain threshold of rats in each group was measured using an acupuncture device before administration and on the day after the last administration.

[0128] The joint circumference was measured using the suture method before administration and on the day after the last administration to observe the swelling of the animal's shoulder joint.

[0129] After the last withdrawal of the drug, rats in each group were euthanized by exsanguination of the abdominal aorta after isoflurane inhalation anesthesia. The skin of the rat model site was cut open, and the right biceps femoris muscle tissue was dissected and harvested. The tissue homogenate was prepared with 0.9% sodium chloride injection at a ratio of 1:9. The contents of TNF-α, IL-6, PGE-2 and TGF-β1 in the biceps femoris muscle tissue were determined by ELISA.

[0130] Experimental results

[0131] (1) Effect on the circumference of shoulder joint swelling

[0132] As shown in Table 8, compared with the normal control group, the swelling circumference of the shoulder joint in the model control group rats was significantly increased before drug administration and on the day after the last drug administration.P ≤ 0.01); Compared with the model control group, except for the control group 10, the circumference of shoulder joint swelling in the other drug-treated groups was significantly reduced on the day after the last administration ( P ≤ 0.05 or P (≤ 0.01), the effects of Zhengqing Fengtongning injection group and Examples 6-8 in reducing the swelling circumference were basically the same, while the effects of Example 5 and Comparative Examples 6-9 were slightly worse, and the effect of Comparative Example 10 in reducing the swelling circumference was the worst.

[0133] Table 8. Effects of anti-inflammatory gel patch on the circumference of shoulder joint swelling in rats ( (n=10)

[0134]

[0135] Note: Compared with the normal control group + P ≤0.05; compared with the model control group * P ≤0.05, ** P ≤0.01.

[0136] (2) Effect on shoulder pain threshold

[0137] As shown in Table 9, compared with the normal control group, the shoulder pain threshold of rats in the model control group was significantly reduced before drug administration and on the day after the last drug administration. P ≤ 0.01); Compared with the model control group, except for the control group 10, the shoulder pain threshold of rats in the other drug administration groups was significantly increased on the day after the last administration ( P ≤ 0.05 or P (≤ 0.01), the pain threshold increase effect of Zhengqing Fengtongning Injection Group and Examples 6-8 is basically the same, the pain threshold increase effect of Example 5 and Comparative Examples 6-9 is slightly worse, and the pain threshold increase effect of Comparative Example 10 is the worst.

[0138] Table 9. Effects of anti-inflammatory gel patch on shoulder joint pain threshold in rats ( (n=10)

[0139]

[0140] Note: Compared with the normal control group ++ P ≤0.01; compared with the model control group * P ≤0.05, ** P ≤0.01.

[0141] (3) Effects on cytokines in the biceps femoris muscle tissue of the shoulder joint

[0142] As shown in Table 10, compared with the normal control group, the levels of TNF-α, IL-6, PGE-2, and TGF-β1 in the tissues of rats in the model control group were significantly increased. P ≤ 0.01); compared with the model control group, the levels of TNF-α, IL-6, PGE-2, and TGF-β1 in the tissues of rats in the drug-treated group were significantly reduced ( P ≤ 0.05 or P ≤ 0.01).

[0143] Table 10 Effects of anti-inflammatory gel patch on cytokines in rat biceps femoris muscle tissue ( (n=10)

[0144]

[0145] Note: Compared with the normal control group ++ P ≤0.01; compared with the model control group * P ≤0.05, ** P ≤0.01.

[0146] This study induced a rat model of frozen shoulder using the continuous complete Freund's adjuvant method, while setting up a positive control group as a control, in order to evaluate and compare the efficacy of Sinomenium acutum extract gel patch on frozen shoulder.

[0147] The observation results are as follows

[0148] (1) Effect on shoulder pain threshold: The shoulder pain threshold of rats in groups 5-8 was significantly higher than that of groups 6-9 in comparison after the last administration, indicating that the addition of wintergreen oil helps to improve the analgesic effect of Sinomenium acutum extract gel patch; the shoulder pain threshold of rats in comparison 10 was the worst, indicating that the analgesic effect of the single wintergreen oil prescription was weaker than that of Sinomenium acutum extract alone or in combination; the shoulder pain threshold of rats in groups 6-8 was basically the same as that of the Zhengqing Fengtongning injection group after the last administration, indicating that the analgesic effect of the corresponding prescription Sinomenium acutum extract gel patch was excellent.

[0149] (2) Effect on shoulder joint swelling circumference: The shoulder joint swelling circumference of rats in groups 5-8 was significantly lower than that of control groups 6-9 the day after the last administration, indicating that the addition of wintergreen oil helps to improve the swelling reduction effect of Sinomenium acutum extract gel patch; control group 10 rats had the worst effect in reducing shoulder joint swelling, indicating that the swelling reduction effect of wintergreen oil alone is weaker than that of Sinomenium acutum extract alone or in combination; the shoulder joint swelling circumference of rats in groups 6-8 was basically the same as that of Zhengqing Fengtongning injection group the day after the last administration, indicating that the swelling reduction effect of the corresponding Sinomenium acutum extract gel patch is excellent.

[0150] (3) Effect of shoulder joint biceps femoris tissue cytokines: The shoulder joint muscle tissue cytokines of rats in groups 5-8 were significantly lower than those in control groups 6-9 the day after the last administration, indicating that the addition of wintergreen oil helps to improve the anti-inflammatory effect of Sinomenium acutum extract gel patch; control group 10 showed the worst reduction in shoulder joint muscle tissue cytokines, indicating that the anti-inflammatory effect of the single wintergreen oil prescription was weaker than that of Sinomenium acutum extract alone or in combination; the shoulder joint muscle tissue cytokines of rats in groups 6-8 were basically the same as those in the Zhengqing Fengtongning injection group the day after the last administration, indicating that the anti-inflammatory effect of the corresponding Sinomenium acutum extract gel patch was excellent.

[0151] Effects of anti-inflammatory gel patch on a rat soft tissue injury model

[0152] A rat soft tissue injury model was established by inducing leg muscle injury in rats using the mechanical impact method. The effects of different administration times of anti-inflammatory gel patches on the rat soft tissue injury model were investigated by detecting the pain threshold and degree of soft tissue injury in rats at different times.

[0153] Seventy-six SPF-grade SD rats, half male and half female, with intact skin and no signs of fracture, were selected after quarantine and divided into a normal control group (n=10) and a model group (n=66). The normal control group received no treatment, while the model group rats were anesthetized with isoflurane inhalation (2%–4% for induction and 0.25%–2% for maintenance) and a soft tissue injury model was induced using the impact method. The day after modeling, 60 rats with significant swelling at the modeling site were randomly divided into 6 groups: the model control group, and groups receiving treatment 1, 2, 3, 4, and 5 as described in Example 6, with 10 animals in each group. Before daily administration, both the Example 6 and blank patches were cut to an area of ​​3.0cm × 3.0cm. The leg muscles of each group were prepared periodically. In the treatment groups, the patches were directly applied to the lateral muscles of the left hind limb, and medical breathable adhesive tape was used to fix the patches to prevent them from falling off. For Example 6, groups 1, 3, and 5 received the patch for 8 hours, 12 hours, and 24 hours respectively, with the patch being removed after treatment. The treatment was administered once daily. For Example 6, groups 2 and 4 received the patch for 8 hours and 12 hours respectively, with the patch being removed after treatment. The treatment was administered twice daily for 7 consecutive days. The normal control group and the model control group received the same area of ​​the blank patch from Example 6 using the same administration method. The degree of soft tissue damage in each group was observed on day 4 of administration and the day after the last administration, and scored according to the standards in the table below. The pain threshold was measured before administration, on day 4 of administration, and the day after the last administration.

[0154] Table 11 Observation Indicators and Scoring Standards for Soft Tissue Injury Degree

[0155]

[0156] Experimental results

[0157] (1) Effects on the degree of soft tissue injury in rats

[0158] As shown in Table 12, compared with the normal control group, the soft tissue injury scores of the model control group were significantly increased before drug administration, on day 4 after drug administration, and on day 7 after drug administration (P ≤ 0.01); compared with the model control group, the soft tissue injury scores of rats in groups 2-5 of Example 6 were significantly reduced on the day after the last drug administration (P ≤ 0.05 or P ≤ 0.01).

[0159] Table 12 Effects of different administration durations on the degree of soft tissue injury in rats in Example 6 ( (n=10)

[0160]

[0161] Note: Compared with the normal control group, ++P ≤ 0.01; compared with the model control group, *P ≤ 0.05, **P ≤ 0.01.

[0162] (2) Effect on the pain threshold of soft tissue in rats

[0163] As shown in Table 13, compared with the normal control group, the soft tissue pain threshold of the model control group was significantly reduced before drug administration, on day 4 after drug administration, and on day 7 after drug administration. P ≤ 0.01); Compared with the model control group, the soft tissue pain threshold of rats in groups 2-5 of Example 6 was significantly increased the day after the last administration ( P ≤ 0.05 or P ≤ 0.01).

[0164] Table 13 Effect of different administration durations on pain threshold in rats in Example 6 ( (n=10)

[0165]

[0166] Note: Compared with the normal control group, ++P ≤ 0.01; compared with the model control group, *P ≤ 0.05, **P ≤ 0.01.

[0167] The above description, in conjunction with specific embodiments, provides a further detailed explanation of this application and should not be construed as limiting the specific implementation of this application to these descriptions. For those skilled in the art, various simple deductions or substitutions can be made without departing from the concept of this application, and all such modifications or substitutions should be considered within the scope of protection of this application.

Claims

1. An external use anti-inflammatory traditional Chinese medicine composition, characterized in that, The composition is prepared from the following components in parts by weight: Sinomenium 30-120 parts, wintergreen oil 0.35-1.4 parts, and menthol 0.3-1.2 parts.

2. The external use anti-inflammatory traditional Chinese medicine composition according to claim 1, characterized in that, The composition is prepared from the following components in parts by weight: Sinomenium 30-120 parts, wintergreen oil 0.35-1.4 parts, and menthol 0.3-1.2 parts.

3. The external use anti-inflammatory traditional Chinese medicine composition according to claim 1 or 2, characterized in that, The wintergreen oil includes a synthetic source and an extracted source; the synthetic source includes synthetic methyl salicylate; the extracted source includes an extract obtained by extracting a plant, and the extract includes any one or more of an extract of Sinopodophyllum emodi, an extract of Sinopodophyllum emodi var. chinense, an extract of Sinopodophyllum emodi var. glaucum, an extract of Amomum villosum, an extract of Ilex chinensis, and an extract of Betula alba.

4. The external use anti-inflammatory traditional Chinese medicine composition according to claim 1 or 2, characterized in that, The content of methyl salicylate in the wintergreen oil is more than 90%.

5. The external use anti-inflammatory traditional Chinese medicine composition according to claim 1 or 2, characterized in that, The Sinomenium is extracted with an aqueous ethanol solution, concentrated, and dried to obtain a Sinomenium extract.

6. Use of the external anti-inflammatory traditional Chinese medicine composition according to any one of claims 1-5 in the preparation of a product for treating osteoarthritis, periarthritis of the shoulder, soft tissue injury, and muscle pain.

7. Use according to claim 6, characterized in that, The product for treating osteoarthritis, periarthritis of the shoulder, soft tissue injury, and muscle pain is in the form of a gel, an ointment, a liniment, or a film.

8. Use according to claim 6, characterized in that, The product for treating osteoarthritis, periarthritis of the shoulder, soft tissue injury, and muscle pain is in the form of a gel.

9. An anti-inflammatory gel patch characterized in that, The product for treating osteoarthritis, periarthritis of the shoulder, soft tissue injury, and muscle pain comprises the external anti-inflammatory traditional Chinese medicine composition according to any one of claims 1-5.

10. The anti-inflammatory gel patch of claim 9, wherein, The gel base comprises a high-molecular backbone material, a tackifier, a humectant, a pH regulator, a crosslinking agent, a crosslinking regulator, a surfactant, a filler, and water.

11. A method of preparing an anti-inflammatory gel patch according to claim 10, wherein The method comprises the following steps: (1) preparing a Sinomenium extract: the Sinomenium is pulverized, extracted with an aqueous ethanol solution, concentrated, and dried to obtain a Sinomenium extract; (2) preparing a water-phase mixture: the crosslinking regulator, the humectant, the emulsifier, and part of the tackifier are added to water, and the Sinomenium extract is added and mixed to obtain a water-phase mixture; (3) preparing an oil-phase mixture: the backbone material, the crosslinking agent, and the filler are added to glycerol and mixed to obtain an oil-phase mixture; (4) preparing a main drug solution: the emulsifier, the wintergreen oil, and the menthol are added to glycerol and stirred to obtain a main drug solution; (5) preparing a pH regulator solution: the pH regulator is dissolved in water; (6) mixing the paste: the oil-phase mixture, the water-phase mixture, and the main drug solution are mixed, and the pH regulator solution and the tackifier are added and mixed to obtain a total paste mixture; (7) coating and cutting: the total paste mixture is uniformly coated and cut to obtain the gel paste.

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