A pharmaceutical composition and its use in external analgesic products

Abstract

The present application relates to the technical field of traditional Chinese medicine composition, and particularly relates to a pharmaceutical composition and application thereof in external analgesic products. The pharmaceutical composition takes Cortex Lycii Radicis and Sarcococca confusa as monarch drugs, is combined with Dragon's Blood, Olibanum, Borneol, and can contain turpentine oil. The present application replaces Herba Artemisiae Scopariae in Gout Pain Tincture prescription with Cortex Lycii Radicis and Sarcococca confusa in combination, and replaces dried ginger with turpentine oil, which has the activities of promoting penetration and anti-inflammatory analgesia. Animal experiments show that the present application has significant effects of detumescence, analgesia and down-regulation of IL-1β, IL-6 and TNF-α on sodium iodate-induced rat knee osteoarthritis model, and the overall efficacy is better than that of the original prescription, and the skin sensitization rate is significantly reduced, so the present application can be used for preparing external drugs for treating osteoarthritis, hyperosteogeny, scapulohumeral periarthritis and rheumatic arthritis.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine composition technology, and in particular to a pharmaceutical composition and its application in topical analgesic products. Background Technology

[0002] Osteoarticular diseases are a class of degenerative conditions caused by various factors, including degeneration and damage to articular cartilage or reactive hyperplasia of the joint margins and subchondral bone. The causes are usually related to genetic factors, genes, and articular cartilage degeneration. Clinically, they mainly manifest as pain, swelling, stiffness, and deformity around the joint; in severe cases, intense pain and significant impairment of joint function can occur. Osteoarticular diseases mainly include inflammatory diseases such as osteoarthritis, rheumatoid arthritis, and frozen shoulder, as well as physiological diseases such as osteoporosis and bone hyperplasia. They are characterized by high incidence, long course, and high recurrence rate, seriously affecting patients' quality of life.

[0003] Gutongling Tincture is a classic external Chinese patent medicine already on the market, listed in Part I of the Chinese Pharmacopoeia. Its prescription consists of: 80g of *Saussurea involucrata*, 110g of dried ginger, 1g of dragon's blood, 5g of frankincense, 5g of myrrh, and 1.5g of borneol, prepared into a 1000ml tincture. Its functions and indications are warming the meridians and dispelling cold, dispelling wind and promoting blood circulation, and relieving pain. It is used for lumbar and cervical spondylosis, osteoarthritis, frozen shoulder, and rheumatoid arthritis. In the existing technology, several studies have been conducted on improving the prescription and enhancing the safety of Gutongling Tincture. Mi Jun et al. (Study on transdermal properties and anti-inflammatory effects of short-term administration of Gutongling Tincture, West China Journal of Pharmaceutical Sciences, 2023, 38(6):665-670) studied the transdermal permeability and anti-inflammatory effects of Gutongling Tincture, and confirmed that it can achieve good transdermal absorption and anti-inflammatory effects with short-term administration (about 1 hour). Its anti-inflammatory mechanism is related to the downregulation of inflammatory factors such as IL-1β, 5-HT, iNOS, and IL-6. However, the study also pointed out that some patients reported adverse reactions such as redness and itching after applying Gutongling Tincture, suggesting that its skin safety still needs to be improved.

[0004] Guan Juanjuan (Master's Thesis, 2014, "Study on Reducing Skin Allergy of Gutongling Tincture with Various Traditional Chinese Medicine Extracts") systematically studied the allergens of Gutongling Tincture through skin allergy experiments. The results showed that *Saussurea involucrata* and *Dracaena cochinchinensis* were likely the main allergens, with an allergy rate as high as 30% in the packaged product. The study also found that dried ginger may have certain anti-allergic and anti-inflammatory effects. This study attempted to reduce the allergy rate by adding anti-allergic components such as baicalin and purslane extract, but this only provided partial relief and failed to fundamentally eliminate the allergens. In addition, Chinese invention patent CN118576685B discloses a preparation method for Gutongling Tincture, which improves the yield of effective components such as *Saussurea involucrata* through microwave extraction and other processes, but still uses the original formula and does not solve the toxicity and allergenicity problems of *Saussurea involucrata*. Chinese invention patent CN118001367A discloses the application of a combination of Gutongling tincture and a detoxifying traditional Chinese medicine composition in the preparation of drugs for treating osteoarthritis. The raw materials of the detoxifying traditional Chinese medicine composition include Gentiana macrophylla and Kangfuxin liquid. This combination can significantly reduce the toxicity of Gutongling tincture and significantly reduce allergic reactions to Gutongling tincture. However, the above schemes are all based on retaining the original prescription (containing Saussurea involucrata), and do not eliminate the toxic principal drug from the source of the prescription. Patients still face the risks of cardiotoxicity and skin allergies.

[0005] It is particularly important to note that the principal ingredient in the Bone Pain Relief Tincture, *Aconitum carmichaelii*, has a complex origin, being the dried tuberous roots of various plants in the genus *Aconitum* of the Ranunculaceae family, not a single species. Its main sources include *Aconitum brachypodum* Diels, *A. brachypodum var. laxiflorum* Fletcher et Lauener, *A. brachypodum var. crispulum* WTWang, *A. nagarum* Stapf var. lasiandrum WTWang, *A. nagarum* Stapf var. heterotrichum* Fletcher et Lauener, *A. pendulum* Busch, and *A. flavum* Hand.-Mazz., among others. Due to the diverse origins and significant differences in the content of active ingredients and the proportion of toxic components among medicinal materials from different regions and sources, it is difficult to uniformly control the quality of medicinal materials, leading to great uncertainty in efficacy evaluation and safety in clinical applications. The main active components of *Artemisia argyi* are diterpenoid alkaloids (such as artemisinin A and aconitine), which, while possessing strong effects in dispelling wind and dampness, relieving pain, also exhibit significant cardiotoxicity and neurotoxicity, with a narrow safety window; overdose can cause arrhythmia and even death. Furthermore, its alkaloid components have been proven to be one of the main skin allergens in bone pain relief tincture. Therefore, how to replace *Artemisia argyi* with low-toxicity, low-allergenic medicinal materials from the source of the prescription, while maintaining or even enhancing the anti-inflammatory and analgesic efficacy of the original prescription and significantly reducing toxic side effects, is a pressing technical problem to be solved in this field.

[0006] Therefore, how to replace Artemisia capillaris with low-toxicity, low-allergenic medicinal materials from the source of the prescription, while maintaining or even enhancing the anti-inflammatory and analgesic efficacy of the original prescription and significantly reducing toxic side effects, is a technical problem that urgently needs to be solved in this field. Summary of the Invention

[0007] To address the aforementioned technical problems, the present invention aims to provide a pharmaceutical composition and its application in topical analgesic products. Addressing the technical issues of existing bone pain relief tincture prescriptions containing the toxic principal ingredient, *Saussurea involucrata*, which causes cardiotoxicity, neurotoxicity, and a high rate of skin sensitization, the present invention provides a pharmaceutical composition and its application in topical analgesic products. This aims to significantly reduce toxic side effects and skin allergic reactions by replacing *Saussurea involucrata* with a low-toxicity, low-sensitization herbal combination from the prescription source, thereby providing a safer and more effective pharmaceutical composition for the topical treatment of osteoarthritis, bone hyperplasia, frozen shoulder, and rheumatoid arthritis.

[0008] To achieve the above-mentioned technical effects, the present invention adopts the following technical solution: In a first aspect, the present invention provides a pharmaceutical composition, wherein the raw material comprises the following components in parts by weight: 30-50 parts of scutellaria barbata, 30-50 parts of ajuga decumbens, 0.5-2 parts of dragon's blood, 2-8 parts of frankincense, 2-8 parts of myrrh, and 0.8-2 parts of borneol.

[0009] As one of the preferred technical solutions, the active pharmaceutical ingredient of the pharmaceutical composition further contains 1 to 4 parts of turpentine oil.

[0010] More preferably, by weight, the following components are present: 40 parts of senna bark, 40 parts of ajuga decumbens, 2 parts of turpentine oil, 1 part of dragon's blood, 2-5 parts of frankincense, 2-5 parts of myrrh, and 1-1.5 parts of borneol.

[0011] As one of the preferred technical solutions, the weight ratio of the senna bark to the ajuga decumbens is preferably 1:1.

[0012] As one of the preferred technical solutions, the dosage form of the pharmaceutical composition is a rubber plaster.

[0013] Secondly, the present invention further provides a method for preparing the above-mentioned pharmaceutical composition, comprising: (1) Grind the bark of Xiangjiapi, Jinjiacao, myrrh and frankincense into coarse powder, mix them well, and extract them by heating and reflux with ethanol of 45% to 55% by volume. Collect the extract, filter it, combine the filtrates and concentrate them into a clear paste. (2) Dissolve the dragon's blood and borneol in ethanol and combine them with the clear paste obtained in step (1); (3) Add turpentine to the mixture obtained in step (2), then add a matrix composed of rubber, zinc oxide, lanolin, petrolatum and gasoline, stir evenly, apply it to the backing material, dry, cover and slice to obtain rubber paste.

[0014] Compared with the prior art, the present invention has the following beneficial effects: Firstly, in terms of prescription composition, this invention uses a synergistic combination of Periploca sepium root bark and Ajuga decumbens as the principal ingredient, replacing Artemisia argyi. Periploca sepium root bark (dried root bark of Periploca sepium Bge., a plant in the Asclepiadaceae family) primarily contains cardiac glycosides (such as ajugol and ajugol glycoside) and C21 steroidal glycosides, as well as 4-methoxysalicylaldehyde and sitosterol. Pharmacological studies have shown that Periploca sepium root bark has anti-inflammatory and immunomodulatory effects; its anti-inflammatory effect may be related to inhibiting the release of inflammatory factors and regulating related signaling pathways; its analgesic effect may be related to the inhibition of the central nervous system. Ajuga decumbens (dried whole herb of Ajuga decumbens Thunb., a plant in the Lamiaceae family) primarily contains diterpenoids (neocrotran and rosinane diterpenoids), steroids, iridoid glycosides, and flavonoids. Modern pharmacological studies have shown that *Ajuga decumbens* possesses anti-inflammatory, antibacterial, and immunomodulatory pharmacological activities. Its anti-inflammatory effect is related to the inhibition of the secretion and release of pro-inflammatory factors such as TNF-α and IL-1β. The combination of these two herbs may synergistically exert anti-inflammatory and analgesic effects by acting on different inflammatory pathways, and neither contains aconitine diterpenoid alkaloids, theoretically eliminating the risk of cardiotoxicity and neurotoxicity caused by aconitine components. Skin allergy tests have shown that replacing the above-mentioned herbs can significantly reduce the sensitization potential of this drug composition.

[0015] Secondly, this invention uses turpentine oil instead of dried ginger. Turpentine oil significantly promotes the transdermal penetration of active ingredients such as Perilla frutescens and Ajuga decumbens by disrupting the lipid bilayer structure of the stratum corneum. Furthermore, it possesses significant anti-inflammatory and analgesic activity (by activating TRP channels to produce an anti-stimulant effect and inhibiting the downregulation of inflammatory factors through the NF-κB pathway), synergistically interacting with the principal drug. Experiments show that replacing turpentine oil with an equal amount of propylene glycol (a purely chemical penetration enhancer) significantly reduces the therapeutic effect, proving that turpentine oil is not only a penetration enhancer, but its pharmacological activity also makes a significant contribution to the overall therapeutic effect. In addition, the amount of turpentine oil used is far lower than the 110g of dried ginger, resulting in a substantial reduction in formulation volume and a significant increase in the concentration of active ingredients. Detailed Implementation

[0016] The following embodiments are only used to more clearly illustrate the technical solutions of the present invention. For those skilled in the art, any equivalent modifications and substitutions to the embodiments described below are also within the scope of the present invention. Therefore, all equivalent transformations and modifications made without departing from the spirit and scope of the present invention should be covered within the scope of the present invention.

[0017] To better illustrate the present invention, numerous specific details are set forth in the following detailed embodiments. Those skilled in the art will understand that the present invention can be practiced without certain specific details. In other embodiments, methods, means, apparatus, and steps well known to those skilled in the art are not described in detail in order to highlight the spirit of the invention.

[0018] Example 1

[0019] This embodiment provides a pharmaceutical composition rubber plaster, the prescription of which is: 40g of Xiangjiapi, 40g of Jinjiacao, 2g of turpentine oil, 1g of Longxuejie, 5g of frankincense, 5g of myrrh, and 1.5g of borneol.

[0020] Its preparation method is as follows: (1) Extraction and concentration: Grind 40g of Xiangjiapi, 40g of Jinjucao, 5g of myrrh, and 5g of frankincense into coarse powder (pass through a 10-mesh sieve) and mix well. Add 8 times the total weight of the medicinal materials in a 50% ethanol solution, heat and reflux twice for 1 hour each time, filter, and combine the filtrates. Concentrate the filtrate under reduced pressure at 60℃ and a vacuum of -0.08MPa to a clear extract with a relative density of 1.18 (measured at 60℃), and set aside for later use.

[0021] (2) Drug dissolution: Dissolve 1g of dragon's blood and 1.5g of borneol in 20ml of anhydrous ethanol, then add 2g of turpentine oil, stir and mix well to obtain a drug mixture.

[0022] (3) Matrix preparation and coating: Take the rubber plaster matrix, which consists of 100g rubber, 120g zinc oxide, 20g lanolin, 10g petrolatum, and 400ml No. 120 solvent gasoline. Cut the rubber into small pieces, put them in a sealed container, add gasoline and soak to swell for 24 hours until the rubber is completely dissolved and forms a gel. Add zinc oxide, lanolin and petrolatum to the gel, stir thoroughly to make a uniform paste. Add the drug mixture obtained in step (2) and the clear paste obtained in step (1) to the above paste, and continue stirring for 30 minutes until uniform. Coat the paste on a cotton backing material, control the thickness to 0.2mm, dry in a 70℃ drying oven, cover with an anti-sticking isolation layer, slice, and the rubber plaster is obtained.

[0023] It should be noted that the above preparation conditions are laboratory conditions. In industrial production, parameters such as solvent dosage, extraction time, and drying temperature can be appropriately adjusted according to the actual production scale. The *Aconitum brachypodum* used in the comparative examples of this invention is *Aconitum brachypodum* Diels.

[0024] Comparative Example 1 This comparative example provides a pharmaceutical composition rubber plaster, the formulation of which is: 80g of Saussurea involucrata, 110g of dried ginger, 1g of dragon's blood, 5g of frankincense, 5g of myrrh, and 1.5g of borneol. This comparative example is the original formulation of the prior art bone pain relief tincture (included in the 2025 edition of the Chinese Pharmacopoeia).

[0025] Preparation method: Referring to Example 1, *Saussurea involucrata*, dried ginger, frankincense, and myrrh were pulverized into coarse powder (passed through a 10-mesh sieve), and extracted by heating and reflux using the same process, then concentrated into a clear extract; separately, *Dracaena cochinchinensis* and borneol were dissolved in ethanol to obtain a solution. The above clear extract and solution were combined and further concentrated, and a rubber plaster matrix was added. The preparation process remained the same as in Example 1 to prepare a rubber plaster.

[0026] Comparative Example 2 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Comparative Example 1 in that: Xiangjiapi is used instead of Xue Shang Yi Zhi Hao, and its prescription composition is: Xiangjiapi 80g, dried ginger 110g, dragon's blood 1g, frankincense 5g, myrrh 5g, borneol 1.5g.

[0027] Preparation method: Referring to Example 1, the licorice bark, dried ginger, frankincense, and myrrh were pulverized into coarse powder, and extracted by heating and reflux according to the same process and concentrated into a clear paste; the subsequent dissolving, mixing, and coating steps were kept the same as in Example 1 to prepare a rubber plaster.

[0028] Comparative Example 3 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Comparative Example 1 in that: Ajuga decumbens is used instead of Artemisia argyi. Its prescription composition is: Ajuga decumbens 80g, dried ginger 110g, dragon's blood 1g, frankincense 5g, myrrh 5g, and borneol 1.5g.

[0029] Preparation method: Referring to Example 1, the ajuga decumbens, dried ginger, frankincense and myrrh were pulverized into coarse powder, and extracted by heating and reflux according to the same process and concentrated into a clear paste; the subsequent dissolving, mixing and coating steps were kept the same as in Example 1 to prepare a rubber plaster.

[0030] Comparative Example 4 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Comparative Example 1 in that: Xiangjiapi and Jingucao replace Xue Shang Yizhihao, and its prescription composition is: Xiangjiapi 40g, Jingucao 40g, dried ginger 110g, dragon's blood 1g, frankincense 5g, myrrh 5g, borneol 1.5g.

[0031] Preparation method: Referring to Example 1, the bark of *Cinnamomum camphora*, *Ajuga decumbens*, dried ginger, frankincense, and myrrh were pulverized into coarse powder, extracted by heating and reflux using the same process, and concentrated into a clear extract; separately, dragon's blood and borneol were dissolved in ethanol (without adding turpentine oil). The above extract and solution were combined and added to the rubber plaster matrix, and the preparation process remained the same as in Example 1 to prepare the rubber plaster.

[0032] Comparative Example 5 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Example 1 in that it lacks turpentine oil. Its prescription consists of: 40g of fragrant bark, 40g of ajuga decumbens, 1g of dragon's blood, 5g of frankincense, 5g of myrrh, and 1.5g of borneol.

[0033] Preparation method: Referring to Example 1, the bark of *Periploca sepium*, *Ajuga decumbens*, frankincense, and myrrh were pulverized into coarse powder, extracted by heating and reflux using the same process, and concentrated into a clear extract; separately, dragon's blood and borneol were dissolved in ethanol (without adding turpentine oil). The above extract and solution were combined and added to the rubber plaster matrix, and the preparation process remained the same as in Example 1 to prepare the rubber plaster.

[0034] Comparative Example 6 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Comparative Example 1 in that turpentine oil is used instead of dried ginger. Its prescription composition is: 80g of Saussurea involucrata, 2g of turpentine oil, 1g of dragon's blood, 5g of frankincense, 5g of myrrh, and 1.5g of borneol.

[0035] Preparation method: Referring to Example 1, *Saussurea involucrata*, frankincense, and myrrh were pulverized into coarse powder, extracted by heating and reflux using the same process, and concentrated into a clear extract; separately, *Dracaena cochinchinensis* and borneol were dissolved in ethanol, and turpentine oil was added and mixed well. The above extract and mixture were combined, and a rubber plaster matrix was added. The preparation process remained the same as in Example 1 to prepare a rubber plaster.

[0036] Comparative Example 7 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Example 1 in that it lacks Ajuga decumbens and increases the amount of Perilla frutescens bark. Its prescription composition is: Perilla frutescens bark 80g, turpentine oil 2g, dragon's blood 1g, frankincense 5g, myrrh 5g, borneol 1.5g.

[0037] Preparation method: Referring to Example 1, the styrax bark, frankincense, and myrrh were pulverized into coarse powder, extracted by heating and reflux according to the same process, and concentrated into a clear paste; the subsequent dissolution, addition of turpentine oil, and matrix coating steps were kept the same as in Example 1 to prepare a rubber plaster.

[0038] Comparative Example 8 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Example 1 in that it lacks Xiangjiapi and increases the amount of Jinjiacao. Its prescription composition is: Jinjiacao 80g, turpentine oil 2g, dragon's blood 1g, frankincense 5g, myrrh 5g, and borneol 1.5g.

[0039] Preparation method: Referring to Example 1, the ajuga decumbens, frankincense, and myrrh were pulverized into coarse powder, extracted by heating and reflux according to the same process, and concentrated into a clear paste; the subsequent dissolution, addition of turpentine oil, and matrix coating steps were kept the same as in Example 1 to prepare a rubber plaster.

[0040] Comparative Example 9 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Example 1 in that the amount of turpentine is different. Its prescription composition is: 40g of fragrant bark, 40g of ajuga decumbens, 1g of turpentine, 1g of dragon's blood, 5g of frankincense, 5g of myrrh, and 1.5g of borneol.

[0041] Preparation method: exactly the same as in Example 1, except that the amount of turpentine added in step (2) is adjusted to 1g to make a rubber paste.

[0042] Comparative Example 10 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Example 1 in that the amount of turpentine is different. Its prescription composition is: 40g of fragrant bark, 40g of ajuga decumbens, 4g of turpentine, 1g of dragon's blood, 5g of frankincense, 5g of myrrh, and 1.5g of borneol.

[0043] Preparation method: exactly the same as in Example 1, except that the amount of turpentine added in step (2) is adjusted to 4g to make a rubber paste.

[0044] Comparative Example 11 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Example 1 in that turpentine is replaced with propylene glycol, a chemical penetration enhancer. The prescription consists of: 40g of scutellaria barbata, 40g of ajuga decumbens, 2g of propylene glycol, 1g of dragon's blood, 5g of frankincense, 5g of myrrh, and 1.5g of borneol.

[0045] Preparation method: Referring to Example 1, extract and concentrate the following herbs: *Periploca sepium*, *Ajuga decumbens*, frankincense, and myrrh; separately, dissolve *Dracaena cochinchinensis* and borneol in ethanol, and add 2g of propylene glycol to replace turpentine. Subsequent preparation processes remain the same as in Example 1 to produce a rubber plaster.

[0046] Comparative Example 12 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Example 1 in that: *Shenjincao* replaces *Jin Gu Cao*, and its prescription composition is: 40g of *Xiangjiapi*, 40g of *Shenjincao*, 2g of turpentine oil, 1g of dragon's blood, 5g of frankincense, 5g of myrrh, and 1.5g of borneol.

[0047] Preparation method: Referring to Example 1, the herbs *Xiangjiapi*, *Shenjincao*, frankincense, and myrrh were pulverized into coarse powder, extracted by heating and reflux according to the same process, and concentrated into a clear paste; subsequent steps were kept the same as in Example 1 to prepare a rubber plaster.

[0048] Comparative Example 13 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Example 1 in that: Clematis chinensis replaces Ajuga decumbens, and its prescription composition is: 40g of Perilla frutescens, 40g of Clematis chinensis, 2g of turpentine oil, 1g of dragon's blood, 5g of frankincense, 5g of myrrh, and 1.5g of borneol.

[0049] Preparation method: Referring to Example 1, the herbs citronella bark, clematis root, frankincense, and myrrh were pulverized into coarse powder, extracted by heating and reflux according to the same process, and concentrated into a clear paste; subsequent steps were kept the same as in Example 1 to prepare a rubber plaster.

[0050] Comparative Example 14 This comparative example provides a pharmaceutical composition rubber plaster, which differs from Comparative Example 1 in that: dried ginger is removed, and its prescription composition is: 80g of Saussurea involucrata, 1g of Dragon's Blood, 5g of Frankincense, 5g of Myrrh, and 1.5g of Borneol.

[0051] Preparation method: Referring to Example 1, Saussurea involucrata, frankincense, and myrrh were pulverized into coarse powder, extracted by heating and reflux using the same process, and concentrated into a clear extract; separately, dragon's blood and borneol were dissolved in ethanol (without adding turpentine oil). The above extract and solution were combined and added to the rubber plaster matrix, and the preparation process remained the same as in Example 1 to prepare the rubber plaster.

[0052] Experimental Example 1: Pharmacodynamic Experiment of a Rat Knee Osteoarthritis Model Induced by Sodium Iodoacetate 1.1 Experimental Animals and Model Construction Experimental animals: SPF-grade male SD rats, weighing 180-220g, were housed in an SPF-grade animal room at a temperature of 22±2℃ and a relative humidity of 50%-60%, with 12 hours of alternating light and dark conditions, and free access to water and food.

[0053] Establishment of a knee osteoarthritis model: Rats were fasted for 12 hours prior to modeling and placed in a special plastic box for gas anesthesia. After anesthesia with 2%–3% isoflurane gas, the right knee of the rats was shaved. The rats were fixed in a supine position on an operating table and disinfected with 75% ethanol. The right knee was flexed to 90°. The midpoint of the patellar ligament was used as the needle insertion point, with the insertion angle parallel to the tibial plateau. Successful puncture was confirmed by a feeling of emptiness under the needle and no blood aspiration. Sodium iodoacetate (1 mg dissolved in 30 μL of physiological saline) was injected into the rat's knee joint cavity using a Hamilton microsyringe. After injection, the needle was slowly withdrawn, and hemostasis was achieved by applying pressure with a cotton ball for 2 minutes. The rats were then subjected to flexion and extension movements of the knee joint to ensure even drug infiltration throughout the joint cavity. In the normal control group, 30 μL of physiological saline was injected into the right knee joint cavity of rats.

[0054] Fourteen days after modeling, four rats from the model group and four rats from the normal control group were randomly selected. Under gas anesthesia, X-rays of the right knee were taken in anteroposterior and lateral views. If the medial joint space of the knee was narrowed and blurred, and the surface of the knee joint was not smooth, it proved that the knee osteoarthritis model was successful.

[0055] 1.2 Grouping and Dosing Rats that successfully developed the model were randomly divided into 17 groups of 10 rats each: normal control group, model control group, Example 1 group, and comparative groups 1-14. The normal control group and model control group did not receive any drugs, while the other groups had the corresponding rubber plaster applied to the hairless area (approximately 2cm × 2cm) on the right knee of the rats.

[0056] The amount of the main medicinal material remained consistent across all treatment groups, at 3.96 g of main medicinal material per kg of body weight. The main medicinal material refers to the principal ingredient in the prescription (i.e., *Periploca sepium*, *Ajuga decumbens*, and *Artemisia scoparia*; other medicinal materials are not counted as principal medicinal materials). Specifically, this includes the total amount of *Periploca sepium* and *Ajuga decumbens* in Example 1, the amount of *Artemisia scoparia* in Comparative Examples 1, 6, and 14, and the corresponding total amount of the principal ingredient in other comparative examples. The treatment was administered once daily for 6 hours each time, for 3 consecutive weeks.

[0057] 1.3 Detection Indicators and Detection Results (1) Measurement of knee joint swelling: The transverse diameter of the right knee joint of rats in each group was measured using an electronic digital vernier caliper in a natural flexion state before modeling, after modeling (before drug administration), and 3 weeks after drug administration. Each measurement was performed by 3 researchers, and the average of the two measurements was taken. Knee joint swelling (%) was calculated according to the following formula: Knee joint swelling (%) = [(knee joint diameter after drug administration - knee joint diameter before modeling) / knee joint diameter before modeling] × 100%.

[0058] The experimental results are shown in Table 1.

[0059] (2) Pain behavioral testing (mechanical withdrawal threshold PWT): The mechanical withdrawal threshold of rats in each group was tested one day before modeling and on the last day of drug administration. For each formal test, the SD rats were placed in a transparent pain testing cage for 10 minutes to allow them to fully adapt to the new environment, stop trembling, and remain completely quiet before the experiment began. Von Frey wire was used to press vertically on the skin between the toes of the SD rat's hind paw, avoiding the central area of ​​the paw, and upward force was applied to make the Von Frey wire bend slightly and hold for about 2 seconds. The stimulation intensity was gradually increased from a minimum of 4g until the rats showed an escape response such as withdrawal or licking their paws. The weight of the Von Frey wire at this time was recorded as the PWT value of that test. Each rat was tested 5 times, with a 1-minute interval between each stimulation. The PWT value of the rat was defined as the number of escape responses in 3 out of 5 tests. The experimental results are shown in Table 1.

[0060] Table 1. Comparison of knee joint swelling and mechanical withdrawal threshold (PWT) in rats after drug administration in each group ( (n=10)

[0061] Note: Compared with the model control group, **P<0.01; compared with Example 1 group, #P<0.05, ##P<0.01; compared with Comparative Example 1 group, △ P<0.05, △△ P<0.01.

[0062] As shown in Table 1, the knee joint swelling of rats in the model control group was significantly increased, and the mechanical withdrawal threshold (PWT) was significantly decreased, showing a highly significant difference compared with the normal control group (P<0.01), indicating that the knee osteoarthritis model was successfully established. After 3 weeks of drug administration, the knee joint swelling of rats in each drug administration group decreased to varying degrees, and the mechanical withdrawal threshold increased. Among them, the knee joint swelling of rats in Example 1 group was significantly lower than that in Comparative Example 1 group (original formula of Gutongling Tincture), and the mechanical withdrawal threshold was significantly higher than that in Comparative Example 1 group, with statistically significant differences (P<0.01), indicating that under the same dosage of the main medicinal materials, the drug composition provided by this invention is significantly better than the original formula of Gutongling Tincture in improving joint swelling and pain.

[0063] Regarding the dosage of turpentine oil, the swelling degree and PWT improvement in Comparative Example 10 (4g turpentine oil) were comparable to those in Example 1 (P>0.05), and both were significantly better than Comparative Example 1 (P<0.01), indicating that turpentine oil can exert excellent therapeutic effects within the dosage range of 2-4g. However, considering that Comparative Example 10 used a relatively large amount of turpentine oil, it would increase raw material costs and cause waste in the industry. Furthermore, high doses of volatile oil can easily lead to softening of the plaster during preparation, affecting the stability of the molding process and the excipient properties of the finished product. Therefore, Example 1 (2g turpentine oil) has greater advantages for industrial production while ensuring efficacy. Comparative Example 2 and Comparative Example 3 used either Persicae Pi or Ajuga decumbens as substitutes for the original principal drug, respectively. Their efficacy indicators were significantly lower than those of Comparative Example 1 (P<0.01), indicating that Persicae Pi and Ajuga decumbens need to be used in combination to achieve synergistic effects. Furthermore, comparative groups 4, 5, 9, and 14 showed significantly better results than comparative group 1 in terms of swelling degree or PWT index, suggesting that optimizing the principal drug combination, adjusting the dosage of turpentine oil, or removing dried ginger can improve the efficacy of the original formula to some extent. Comparative group 11 used propylene glycol instead of turpentine oil, and although its efficacy was better than comparative group 1, it was not as good as that of example group 1, further demonstrating that turpentine oil is not only a penetration enhancer but also possesses unique pharmacological activity.

[0064] (3) Determination of IL-1β, IL-6, and TNF-α content in joint lavage fluid: After drug administration, rats in each group were fully anesthetized by intraperitoneal injection of 1% sodium pentobarbital (50 mg / kg). The rats were fixed on the operating table, and the skin of both knee joints was routinely disinfected. A 1 mL sterile syringe connected to a 27G needle was inserted vertically into the joint cavity of both knees of the rats, and 0.5 mL of pre-cooled sterile saline was slowly injected into each knee joint cavity. The joint cavity was gently pressed 3-5 times to ensure that the lavage fluid came into full contact with the intra-articular tissues. Then, the lavage fluid was slowly aspirated under negative pressure to recover as much lavage fluid as possible. The recovered joint lavage fluid was stored on ice and centrifuged at 3000 r / min for 15 min at 4℃. The supernatant was separated, and the IL-1β, IL-6, and TNF-α content was detected by ELISA. The experimental results are shown in Table 2.

[0065] Table 2 Comparison of inflammatory factor levels in joint lavage fluid of rats in different groups ( (n=10, pg / mL)

[0066] Note: Compared with the model control group, **P<0.01; compared with Example 1 group, #P<0.05, ##P<0.01; compared with Comparative Example 1 group, △ P<0.05, △△ P<0.01.

[0067] As shown in Table 2, the levels of inflammatory factors IL-1β, IL-6, and TNF-α in the joint lavage fluid of rats in the model control group were significantly increased, indicating a severe local inflammatory response in the joints. After drug administration, the levels of inflammatory factors in each treatment group were reduced. Specifically, the levels of the above three inflammatory factors in Example 1 group were significantly lower than those in Comparative Example 1 group (the original formula of Gutongling Tincture), with statistically significant differences (P<0.01), indicating that the drug composition of the present invention has a significantly better effect on inhibiting joint inflammation than the original formula. The levels of each inflammatory factor in Comparative Example 10 group were similar to those in Example 1 group, with no statistical difference, further confirming the effectiveness of the optimal dosage range of turpentine oil. The anti-inflammatory effect of Comparative Example 14 group after removing dried ginger was better than the original formula, suggesting that removing dried ginger may help alleviate the local inflammatory pathological state. Comparative Example 11 group used propylene glycol instead of turpentine oil, and its anti-inflammatory effect was significantly weaker than that of Example 1 group, indicating that turpentine oil plays an important role in the anti-inflammatory efficacy of the formula.

[0068] In summary, based on the experimental results in Tables 1 and 2, Example 1 group showed superior therapeutic effects compared to Comparative Example 1 (original formula) in reducing knee joint swelling, increasing the pain threshold, and downregulating the levels of intra-articular inflammatory factors IL-1β, IL-6, and TNF-α, with statistically significant differences. Comparative analysis showed that the combined use of Xiangjiapi and Jinjiacao (Example 1) was more effective than single-drug use (Comparative Examples 7 and 8), indicating a synergistic effect (P<0.01). Furthermore, the addition of turpentine oil significantly enhanced the efficacy, and its effect was superior to that of the chemical penetration enhancer propylene glycol alone. Experimental data also showed that Comparative Example 10 (4g turpentine oil) had comparable efficacy to Example 1 group (P>0.05), and Comparative Examples 4, 5, 9, and 10 all showed varying degrees of improvement compared to the original formula (P<0.01), indicating that the present invention achieves a significant improvement in efficacy through the synergistic effect of multiple components.

[0069] Experiment Example 2: Comparative Experiment on Skin Allergies 2.1 Experimental Objective This experiment aims to observe the differences in skin allergy caused by the pharmaceutical composition of the present invention (Example 1) compared with the original prescription of Bone Pain Relief Tincture (Comparative Example 1) and the prescription in Comparative Example 14, and to observe the safety of the modified prescription.

[0070] 2.2 Laboratory Animals and Grouping Experimental animals: Hartley white guinea pigs, half male and half female, weighing 250-300g, were used for experiments after acclimatization for one week.

[0071] Grouping: The guinea pigs were randomly divided into 6 groups of 10 each, as follows: Blank control group (no drugs were given); Blank matrix group (given a blank rubber plaster matrix without drugs); Example 1 group (the rubber plaster of Example 1 of the present invention was applied); Comparative Example 1 (administered the original prescription of Bone Pain Relief Tincture rubber plaster, Comparative Example 1); Comparative Example 14 (administered the original formula with dried ginger rubber plaster, Comparative Example 14); Positive control group (administered 2,4-dinitrochlorobenzene DNCB acetone solution).

[0072] 2.3 Experimental Methods Induced sensitization: Hair was removed from the left side of the guinea pig's back in a 3cm x 3cm area 24 hours before the experiment. On days 0, 7, and 14, the test substance for each group was applied to the hair-removed area, fixed with non-irritating adhesive tape for 6 hours, and then removed. The residue was washed off with warm water. The blank control group was not treated with any substance, the blank matrix group was treated with blank matrix, and the positive control group was coated with 1% DNCB acetone solution.

[0073] Challenge exposure: On day 28 (14 days after the last sensitization), hair was removed from the right side of the guinea pig's back in a 2cm x 2cm area. The test substance for each group was applied to the hair-removed area on the right side, fixed for 6 hours, and then removed and washed off. Skin reactions were observed and recorded at 6h, 24h, 48h, and 72h after challenge.

[0074] The skin reaction scoring criteria are as follows: 0 points = no response; 1 point = erythema; 2 points = erythema and edema; 3 points = erythema, edema, and papules or vesicles; 4 points = significant edema or necrosis.

[0075] Sensitization positivity criteria: Animals with a skin reaction score ≥1 at 24 or 48 hours post-provocation are considered sensitized positively. Sensitization rate (%) = (Number of sensitized positive animals / Total number of animals in the group) × 100%.

[0076] 2.4 Dosage setting To scientifically compare the differences in sensitization among the different prescription groups, the following dosage control strategy was adopted in this experiment: (1) Example 1 and Comparative Example 1: The amount of main medicinal material in the control group was kept consistent, which was 3.96 g of main medicinal material / kg body weight; (2) Comparative Example 14: The content of the main sensitizing component, Artemisia argyi, was controlled to be consistent with that of Comparative Example 1 (3.96 g / kg) to investigate the effect of removing dried ginger on the sensitization of the original prescription.

[0077] 2.5 Experimental Results The results of the skin allergy test in each group of guinea pigs are shown in Table 3.

[0078] Table 3 Results of skin allergy test in each group of guinea pigs (n=10)

[0079] The above experimental results show that the guinea pigs in the positive control group exhibited extreme sensitization reactions, with a sensitization rate of 100%, indicating the reliability of the experimental method. No obvious allergic reactions were observed in the blank control group and the blank matrix group. The sensitization rate in Comparative Example 1 (original formula, Gu Tong Ling Tincture) was 40%, showing mild sensitization. This result indicates that the original formula has a high clinical risk of skin sensitization, with its main sensitizing component being *Saussurea involucrata*. The sensitization rate in Comparative Example 14 (original formula without dried ginger) increased to 60%, and the average reaction values ​​at each time point were significantly higher than those in Comparative Example 1, suggesting that under the same dosage of the sensitizing agent *Saussurea involucrata*, removing dried ginger leads to a significant increase in the sensitization rate. This result objectively shows that dried ginger does indeed play a role in inhibiting or antagonizing the sensitization of *Saussurea involucrata* in the original formula, and removing dried ginger exposes a stronger sensitization risk from *Saussurea involucrata*.

[0080] The sensitization rate in Example 1 was 10%, consistent with the blank matrix group, and significantly lower than that in Comparative Example 1 and Comparative Example 14. This data comparison indicates that although dried ginger has a certain inhibitory effect on sensitization in the original formula (the sensitization rate increases after removal), this invention effectively avoids the risks posed by the main allergen by replacing Artemisia capillaris with Persicae and Ajuga decumbens. This technical solution significantly reduces the incidence of skin allergic reactions at the same dosage, demonstrating the clear safety advantages of the new formulation.

[0081] In summary, the pharmaceutical composition provided by this invention, by replacing Artemisia capillaris and Ajuga decumbens with Artemisia scoparia, replacing dried ginger with turpentine oil, and optimizing the proportions of each component, not only achieves a significant improvement in anti-inflammatory and analgesic effects, making it more effective than the original formula Bone Pain Relief Tincture, but also fundamentally reduces skin allergic reactions caused by the toxic principal drug, significantly improving the safety of clinical use. This provides a new, safe, and reliable topical medication option for the treatment of osteoarthritis.

[0082] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the present invention, and all such modifications and substitutions should be covered within the scope of the claims of the present invention. Technical aspects, shapes, and structures not described in detail in this invention are all well-known technologies.

Claims

1. A pharmaceutical composition, characterized in that, The active pharmaceutical ingredient of the pharmaceutical composition, by weight, consists of the following components: 30-50 parts of licorice root bark, 30-50 parts of ajuga decumbens, 0.5-2 parts of dragon's blood, 2-8 parts of frankincense, 2-8 parts of myrrh, and 0.8-2 parts of borneol.

2. The pharmaceutical composition according to claim 1, characterized in that, The weight ratio of the herb bark to the herb is 1:

1.

3. The pharmaceutical composition according to claim 1, characterized in that, The active pharmaceutical ingredient in the pharmaceutical composition also contains 1 to 4 parts of turpentine oil.

4. The pharmaceutical composition according to claim 3, characterized in that, The amount of turpentine used is 2 parts.

5. The pharmaceutical composition according to claim 1, characterized in that, The active pharmaceutical ingredient of the pharmaceutical composition, by weight, consists of the following components: 40 parts of licorice bark, 40 parts of ajuga decumbens, 1 part of dragon's blood, 5 parts of frankincense, 5 parts of myrrh, 1.5 parts of borneol, and 2 parts of turpentine oil.

6. The pharmaceutical composition according to any one of claims 1 to 5, characterized in that, The pharmaceutical composition is prepared as a topical preparation, which is a rubber plaster.

7. The use of the pharmaceutical composition according to any one of claims 1 to 5 in the preparation of a topical analgesic product.

8. The application according to claim 7, characterized in that, The applications include their use in the preparation of medicaments for treating osteoarthritis, bone hyperplasia, frozen shoulder, or rheumatoid arthritis.

9. A method for preparing a pharmaceutical composition according to any one of claims 3 to 5, characterized in that, Includes the following steps: (1) Grind the bark of Xiangjiapi, Jinjiacao, myrrh and frankincense into coarse powder, mix them well, and extract them by heating and reflux with ethanol of 45% to 55% by volume. Collect the extract and concentrate it into a clear paste. (2) Dissolve the dragon's blood and borneol in ethanol and combine them with the clear paste obtained in step (1); (3) Add turpentine to the mixture obtained in step (2), then add the matrix required for preparing rubber paste, stir evenly, and apply it to the backing material to obtain the final product.

10. The preparation method according to claim 9, characterized in that, The matrix consists of rubber, zinc oxide, lanolin, petrolatum, and gasoline.

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