Film-forming gel capable of dispelling wind and eliminating arthralgia as well as preparation method and application of film-forming gel

Abstract

The embodiment of the invention discloses a wind-dispelling and arthralgia-removing film-forming gel as well as a preparation method and application thereof. The wind-dispelling and arthralgia-removing film-forming gel is prepared from, by mass, 0.1-0.9 part of medicine, 0.1-0.5 part of MC-400, 0.1-0.5 part of PVA1788, 0.1-0.9 part of glycerol, 1-3 parts of absolute ethyl alcohol and the balance distilled water, and the medicine is prepared from stems and leaves of aconitum carmichaeli, herba ephedrae, astragalus membranaceus, radix paeoniae alba and honey-fried licorice roots. The traditional discarded aconite stems and leaves are used as raw materials, monkshood as a monarch drug in ancient classical famous formula aconite decoction with remarkable curative effect on clinical treatment of rheumatoid arthritis is replaced by the aconite stems and leaves to relieve pungent heat and fierce property and reduce toxic and side effects, the film-forming gel with remarkable anti-inflammatory and analgesic effects is developed, and the film-forming gel is good in safety, convenient to use and free of toxic and side effects. The waste of the aconite stem and leaf resources is turned into wealth, and the method is of great significance to development and utilization of the aconite stem and leaf resources.

Description

Technical Field

[0001] The embodiments of the present invention relate to the technical field of gel preparations, and specifically to a film-forming gel for dispelling wind and numbness, and a preparation method and application thereof. Background Art

[0002] Rheumatoid arthritis (RA) is an autoimmune disease characterized by polyarticular, symmetrical, and aggressive arthritis of the hands and feet. This can cause joint deformities and loss of function, significantly impacting patients' daily lives. Currently, Western medications used to treat RA primarily include glucocorticoids, nonsteroidal anti-inflammatory drugs (NSAIDs), and anti-rheumatic drugs (DMARDs). However, long-term use can cause numerous adverse reactions, including nausea, infections, and liver and kidney damage. Therefore, there is an urgent need to identify alternative medications with improved efficacy and fewer side effects. Traditional Chinese medicine (TCM) has been used to treat RA and related rheumatic diseases for thousands of years, with remarkable efficacy. Wutou decoction, originating from the Golden Chamber, consists of five herbs: Chuanwu (Aconite), Ephedra, Paeonia lactiflora, Astragalus, and Zhigancao (Glycyrrhiza uralensis). It states that "Wutou decoction is the main treatment for pain and inability to bend or stretch joints." It is now commonly used to treat rheumatic diseases with good results. However, the potent and toxic nature of the main ingredient, Aconitum, limits its use.

[0003] Aconitum carmichaelii (Aconitum carmichaelii Debx.), a renowned Sichuan native medicinal herb, is used primarily for its mother root (Aconiti Radix) and processed daughter root (Aconiti Radix). While its stems and leaves are abundant in biomass, their application is limited, resulting in significant resource waste. Studies have reported that Aconitum carmichaelii stems and leaves are significantly effective in treating rheumatoid arthritis and exhibit less acute toxicity than Aconiti Radix and Aconiti Radix. The total alkaloid content in Aconitum carmichaelii stems and leaves is approximately half that of its underground parts. Our previous research found that Aconitum carmichaelii stems and leaves are rich in aporphine alkaloids and alcohol-amine-type diterpenoid alkaloids, while the content of the more toxic diester-type diterpenoid alkaloids is significantly lower than in other parts (Aconiti Radix and Aconiti Radix), demonstrating the significant potential for the development of Aconitum carmichaelii stems and leaves as a medicinal resource. In recent years, drug delivery systems have garnered significant attention, with transdermal drug delivery rapidly gaining popularity. Compared to traditional oral drug delivery, transdermal drug delivery offers advantages such as avoiding first-pass effects and enhancing patient compliance. Traditional transdermal drug delivery formulations, such as patches, are susceptible to allergic reactions and have poor breathability, while creams can leave a greasy feeling upon application. Film-forming formulations, primarily composed of drugs, polymers, plasticizers, solvents, and excipients, are a highly effective form of drug delivery, serving as reservoirs for sustained release. Summary of the Invention

[0004] Earlier, this research team replaced the processed Chuanwu in the Aconitum decoction with Aconitum stems and leaves to create a homemade wind-clearing and anti-bi wash, demonstrating the enormous potential for development of Aconitum stems and leaves as a medicinal resource. This study further improved the dosage form by mixing the total alkaloids from Aconitum stems and leaves with extracts from the other four herbs in the Aconitum decoction to create a film-forming gel formulation. Compared to the wash, this formulation is easier to use and has good compliance. The anti-inflammatory and analgesic effects of the gel were studied using the acetic acid writhing model and the AIA rheumatoid arthritis model, and its safety was evaluated using a New Zealand rabbit skin irritation test. This study will provide a reference for the exploration of medicinal resources and product development of Aconitum stems and leaves, and contribute to the high-value utilization of the "non-medicinal parts" of traditional Chinese medicine.

[0005] In order to achieve the above objectives, the embodiments of the present invention provide the following technical solutions:

[0006] According to the first aspect of the embodiments of the present invention, the present invention provides a film-forming gel for dispelling wind and relieving arthritis, which comprises 10 parts by mass of the film-forming gel for dispelling wind and relieving arthritis, including: 0.1-0.9 parts of the drug, 0.1-0.5 parts of MC-400, 0.1-0.5 parts of PVA1788, 0.1-0.9 parts of glycerol, 1-3 parts of anhydrous ethanol, and the remainder being distilled water, wherein the drug is prepared using the stems and leaves of Aconitum, Ephedra, Astragalus, White Peony Root and Radix Glycyrrhizae as raw materials.

[0007] Furthermore, it comprises: 0.5 parts of drug, 0.3 parts of MC-400, 0.1 parts of PVA1788, 0.5 parts of glycerol, 1 part of anhydrous ethanol, and the balance is distilled water.

[0008] Furthermore, the preparation method of the drug is as follows:

[0009] The crude powder of Aconitum stem and leaf was added into the liquid at a ratio of 20 times the amount of 0.1 mol·L -1 After soaking in hydrochloric acid solution for 1 hour, the mixture was extracted at 70°C for 3 hours, twice, and the extract was concentrated and adjusted to pH 3 using hydrochloric acid. The extract was then loaded onto a 001×12 type cation exchange resin for enrichment and purification, thereby obtaining the total alkaloids from the stems and leaves of Aconitum carmichaelii.

[0010] Separately, four herbs, namely, ephedra, astragalus, white peony root, and glycyrrhiza uralensis, were mixed in a mass ratio of 1:1:1:1. Six times the amount of the mixed herbs was added to 50% ethanol and soaked for 30 minutes. The mixture was then condensed and refluxed for three times. The extract was concentrated under vacuum at 60°C to a concentration of 0.2 g of the raw herbs per mL.

[0011] The total alkaloids from the stems and leaves of Aconitum carmichaelii were mixed with the extracts of the other four medicinal herbs at a ratio of 1:3 and concentrated to a crude drug amount of 15 g·mL. -1 The obtained medicine is stored in a -4°C refrigerator.

[0012] According to a second aspect of the embodiments of the present invention, the present invention provides a method for preparing the film-forming gel for dispelling wind and relieving numbness as described in any of the above items, the method comprising: weighing MC-400, PVA1788 and propylene glycol according to the prescribed amount, adding part of the distilled water to swell and stir overnight, adding the prescribed amount of anhydrous ethanol and the drug to the gel solution in sequence while stirring, stirring to disperse them evenly, and finally adding the remaining distilled water and mixing them evenly.

[0013] Furthermore, the mass of the partial distilled water accounts for 50-60% of the total mass of the distilled water.

[0014] According to a third aspect of the embodiments of the present invention, the present invention provides use of the Qufengchubi film-forming gel as described above in the preparation of a medicament for rheumatoid arthritis.

[0015] The embodiments of the present invention have the following advantages:

[0016] This study used traditionally discarded Aconitum stems and leaves as raw material, replacing the main ingredient, Chuanwu (Aconiti Radix), in the classic ancient prescription for the treatment of rheumatoid arthritis, Aconitum decoction, with Aconitum stems and leaves to mitigate its pungent and hot properties and reduce its toxic side effects, effectively transforming waste Aconitum stem and leaf resources. This study successfully developed a film-forming gel with significant anti-inflammatory and analgesic properties, which is safe and easy to use, and is of great significance for the development and utilization of Aconitum stem and leaf resources. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] To more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for the embodiments or the description of the prior art. Obviously, the drawings described below are merely exemplary, and those skilled in the art can derive other implementation drawings based on the provided drawings without inventive effort.

[0018] Figure 1 The measurement results of rat toe swelling provided by the present invention;

[0019] Figure 2 The arthritis index score after adjuvant rheumatoid arthritis is induced in SD rats provided by the present invention;

[0020] Figure 3 The measurement results of inflammatory factors provided by the present invention;

[0021] Figure 4 This is the H&E staining picture provided by the present invention;

[0022] Figure 5 The present invention provides the skin irritation results of New Zealand rabbits after repeated administration. DETAILED DESCRIPTION

[0023] The following describes the implementation of the present invention using specific embodiments. Those skilled in the art will readily understand the other advantages and benefits of the present invention from the disclosure herein. Obviously, the embodiments described are only a portion of the present invention, not all of it. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without inventive effort are intended to fall within the scope of protection of the present invention.

[0024] 1. Materials and main instruments

[0025] 1 Animal

[0026] SPF Kunming mice, weighing 18–22 g, half male and half female; standard New Zealand rabbits, weighing 2.0–2.5 kg, half male and half female. Both animals were provided by Chengdu Dashuo Laboratory Animal Co., Ltd., license number: SCXK (Sichuan) 2020-0030. All animals were acclimated for one week prior to the formal experiments. The required environment was: temperature 22 ± 2°C, relative humidity 60 ± 5°C, and a standard 12-h light-dark cycle. All animal experiments were approved by the Ethics Committee of Chengdu University of Traditional Chinese Medicine (No. 2024044).

[0027] 2. Aconitum stem and leaf raw materials

[0028] Stems and leaves of Aconitum carmichaelii were collected from Jiangyou City, Sichuan Province, and identified as Aconitum carmichaelii Dex. by Associate Professor Gao Jihai of Chengdu University of Traditional Chinese Medicine. The specimen (CW-20200901) is currently housed in the "1002 Laboratory for Drug Resource Discovery and Recycling of Traditional Chinese Medicine Resources," State Key Laboratory of Southwest Specialty Traditional Chinese Medicine Resources, Chengdu University of Traditional Chinese Medicine.

[0029] 3 Drugs and reagents

[0030] Sodium carboxymethylcellulose (batch number: 9004-32-4) was purchased from Chengdu Kelong Chemical Co., Ltd.; carbomer (batch number: 013958826) was purchased from Shanghai Titan Technology Co., Ltd.; methylcellulose-400 (batch number: 2017052201) was purchased from Chengdu Kelong Chemical Co., Ltd.; gelatin (batch number: 9000-70-8) was purchased from Zhenghong Biotechnology Co., Ltd.; polyvinyl alcohol 1788 (batch number: 2023042701) was purchased from Chengdu Kelong Chemical Co., Ltd.; hydroxypropyl methylcellulose (batch number: K2115138) was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; polyvinylpyrrolidone (batch number: 9003-39-8) was purchased from Chengdu Kelong Chemical Co., Ltd. Co., Ltd.; hydroxypropyl cellulose (batch number: RH492137) was purchased from Shanghai Yien Chemical Technology Co., Ltd.; anhydrous ethanol (batch number: 2024011601) was purchased from Chengdu Kelong Chemical Co., Ltd.; glycerol (batch number: 20160906) was purchased from Chengdu Jinshan Chemical Reagent Co., Ltd.; triethyl citrate (batch number: 202107209) was purchased from Beijing Mairida Technology Co., Ltd.; polyethylene glycol 400 (batch number: RH519770) was purchased from Shanghai Yien Chemical Technology Co., Ltd.; propylene glycol (batch number: 2021120101) was purchased from Chengdu Kelong Chemical Co., Ltd.; and complete Freund's adjuvant was purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd. ELISA kits (IL-1β: Wuhan Yilairui Biotechnology Co., Ltd., TNF-α: Shanghai ELISA Biotechnology Co., Ltd., IL-6: Shanghai ELISA Biotechnology Co., Ltd.) were used.

[0031] 4 Main instruments

[0032] Rapid TA+ texture analyzer (Shanghai Tengba Instrument Technology Co., Ltd.); BTF-1200C-S viscometer (Shanghai Fangrui Instrument Co., Ltd.); Model 101 electric blast drying oven (Shanghai Yiheng Scientific Instrument Co., Ltd.); BCE2224-1CCN electronic balance (Sartorius Scientific Instruments (Beijing) Co., Ltd.); PHS-2F pH meter (Shanghai Yidian Scientific Instrument Co., Ltd.); ReadMax1200 full-wavelength absorbance microplate reader (Shanghai Flash Biotechnology Co., Ltd.); PV-200 toe swelling meter (Chengdu Taimeng Software Co., Ltd.).

[0033] 2. Data Processing

[0034] Data are expressed as mean ± standard deviation SPSS 25.0 software was used to perform one-way analysis of variance for comparisons among the groups, and P < 0.05 indicated that the differences were statistically significant.

[0035] Example 1 Preparation of a film-forming gel for dispelling wind and removing numbness

[0036] 1 Extract (drug) preparation

[0037] The crude powder of Aconitum stem and leaf was added into the liquid at a ratio of 20 times the amount of 0.1 mol·L -1 Soak in hydrochloric acid solution for 1 hour, extract at 70℃, extract for 3 hours, extract twice, concentrate the extract and load onto 001×12 type cation exchange resin for enrichment and purification. Adjust the sample solution to pH=3 with hydrochloric acid, and the concentration is 0.3g·mL -1 The sample volume was 0.5 BV, 3 BV ultrapure water was used for impurity removal, and then 4 BV of 3 mol·L -1 Elution was performed with ammonia-95% ethanol to enrich and purify the total alkaloids, thus obtaining the total alkaloids from the stems and leaves of Aconitum. 80 g each of the four medicinal materials of Ephedra, Astragalus, White Peony Root, and Radix Glycyrrhizae were placed in the same container, and 50% ethanol, 6 times the amount of the mixed medicinal materials, was added. After soaking for 30 minutes, the extraction was performed under condensation reflux for 3 times, the first time for 1 hour, the second and third times for 0.5 hours each, and the three extracts were combined. The extracts were concentrated under vacuum conditions at 60°C to a crude drug amount of 0.2 g per 1 mL. Finally, the total alkaloids from the stems and leaves of Aconitum were mixed with the extracts of the other four medicinal materials in a crude drug amount of 1:3 and concentrated to a crude drug amount of 15 g·mL -1 The obtained extract (drug) is stored in a -4°C refrigerator.

[0038] 2. Preparation method of film-forming gel

[0039] Weigh the gel matrix, film-forming agent and plasticizer according to the prescribed amount, add water to swell and stir overnight, add the prescribed amount of ethanol and drug to the gel in sequence after stirring evenly, stir to make it evenly dispersed, and finally add water to a sufficient amount to obtain a film-forming gel, which is then packaged in transparent, sealed glass bottles.

[0040] 3 Formulation screening method

[0041] In order to develop a film-forming gel that is easy to use, has good therapeutic effects and good appearance, a single-factor investigation method was adopted. Taking the appearance properties, film-forming time, viscosity, pH, tensile strength and elongation at break of the Qufengchubi film-forming gel as indicators, the single-factor investigation method combined with orthogonal experiments were used to investigate the type and dosage of the Qufengchubi film-forming gel matrix, the type and dosage of the film-forming agent, the type and dosage of the plasticizer, the ethanol concentration and the drug loading amount to determine the optimized formula.

[0042] Table 1 Film-forming gel materials

[0043]

[0044] 3.1 Drying time

[0045] 0.2 g of film-forming gel was placed on a plastic culture dish at 35°C, and the drying time was evaluated by placing a cover glass on the film. If the cover glass was clean and free of foreign matter after removal, the film was considered dry, and the drying time was recorded. Repeat this test three times.

[0046] 3.2 pH measurement

[0047] According to the 2020 edition of the "Chinese Pharmacopoeia" General Chapter 0631, take about 1g of Qufengchubi film-forming gel, add 20mL of water, stir for 30min, use a pH meter to measure, measure each sample 3 times, and record the data.

[0048] 3.3 Viscosity

[0049] The viscosity of the preparation was measured using a viscometer. Each sample was measured three times, and the average and standard deviation were recorded.

[0050] 3.4 Appearance

[0051] The film-forming gel was evaluated by scoring the appearance or properties of the film. 0.5 mL of the film-forming gel was accurately measured and evenly applied to a 4 cm × 4 cm glass plate. After natural drying, the gel and the film formed were evaluated according to Table 2.

[0052] Table 2 Film-forming gel appearance scoring table

[0053]

[0054] 3.5 Mechanical properties (tensile strength, elongation at break)

[0055] 0.5 g of the film-forming gel was placed on a plastic medium at 35°C and allowed to dry to form a film. A 1×5 cm sample was cut from each film-forming gel film for measuring its mechanical properties.

[0056] 4 Screening results of preparation process of Qufengchubi film-forming gel

[0057] 4.1 Investigation of matrix types

[0058] Four matrices, MC-400, gelatin, CMC-Na, and carbomer, were investigated. The experiments revealed that MC-400 and CMC-Na mixed well with the drug and formed a film. However, gelatin swelled unevenly with the drug solution, resulting in a jelly-like appearance and making it unsuitable as a gel matrix. Carbomer, which requires pH adjustment with triethanolamine for dissolution, was too viscous and essentially non-flowing when mixed with the drug. The resulting matrix also became turbid, making it unsuitable as a gel matrix. Compared to CMC-Na, MC-400 exhibited moderate fluidity after mixing. CMC-Na was too viscous, exhibited poor fluidity, and was susceptible to mold. Therefore, MC-400 was selected as the gel matrix, and further investigation was conducted into its optimal dosage.

[0059] 4.2 Investigation of substrate dosage

[0060] The dosage of MC-400 was investigated. Different amounts of MC-400 (0.1g, 0.2g, 0.3g, 0.4g, and 0.5g) were added to a formula (0.2g PVA1788, 0.5g PEG400, 1.5g drug, 1g anhydrous ethanol). The viscosity of the gel increased with increasing matrix dosage. When the MC-400 dosage exceeded 0.3g, the excessive viscosity resulted in an uneven film-forming gel with lumps and difficulty forming a well-formed film, affecting the appearance and spreadability of the film-forming gel. Therefore, the matrix dosage was set at 0.3g (see Table 3).

[0061] Table 3 Investigation of the dosage of film-forming gel matrix

[0062]

[0063] 4.3 Investigation of film-forming agent types

[0064] The effects of four film-forming agents, HPC, PVP, PVA1788, and HPMC-400, on film-forming gel were investigated. It was found that the gel matrix became turbid after the addition of HPC and HPMC-400, while the PVA and PVP gel matrices were transparent and colloid-like. However, the PVP matrix had excessive fluidity and easily flowed away when applied topically, resulting in a poor user experience. PVA1788 had a good appearance and moderate fluidity, so PVA1788 was selected as the subsequent film-forming agent for investigation.

[0065] 4.4 Investigation of film-forming agent dosage

[0066] The dosage of PVA1788 was investigated. Different amounts of PVA1788 (0.1g, 0.2g, 0.3g, 0.4g, and 0.5g) were added to a formulation (0.3g MC-400, 0.5g PEG400, 1.5g drug, and 1g anhydrous ethanol). As the amount of film-forming agent increased, the viscosity increased and the film formed became uneven, with lumpy particles, when the amount of PVA1788 exceeded 0.2g. Therefore, the dosage of film-forming agent for the gel was determined to be 0.2g (see Table 4).

[0067] Table 4 Investigation on the dosage of film-forming gel film-forming agent

[0068]

[0069] 4.5 Plasticizer Type Screening

[0070] The effects of four film-forming agents—glycerol, propylene glycol, PEG400, and TEC—on the properties and film-forming time of the gels were investigated. The results are shown in Table 5. Compared with the group without plasticizers, the addition of propylene glycol and TEC significantly affected the properties of the film-forming gels, resulting in uneven films with cracks. The addition of glycerol and PEG400 resulted in a uniform film, but the tensile strength decreased, while the elongation at break increased. This indicates that the addition of plasticizers improves film toughness. The addition of glycerol plasticizer resulted in better film performance than PEG400, so glycerol was selected as the plasticizer for the film-forming gels.

[0071] Table 5 Investigation of types of film-forming gel plasticizers

[0072]

[0073] 4.6 Plasticizer dosage screening

[0074] The dosage of glycerol was investigated. Different amounts of glycerol (0.1g, 0.3g, 0.5g, 0.7g, and 0.9g) were added to a formulation (0.2g PVA1788, 0.3g MC-400, 1.5g drug, and 1g anhydrous ethanol). As the amount of plasticizer increased, exceeding 0.5g resulted in increased viscosity and uneven film formation, with the presence of lumpy particles. The mechanical properties of the plasticizer were best improved at a glycerol dosage of 0.5g. Therefore, the dosage of 0.5g of glycerol was used (see Table 6).

[0075] Table 6 Investigation of the dosage of film-forming gel plasticizer

[0076]

[0077]

[0078] 4.7 Anhydrous ethanol dosage screening

[0079] The dosage of anhydrous ethanol was investigated. Different amounts of anhydrous ethanol (1 g, 1.5 g, 2 g, 2.5 g, and 3 g) were added to the formula (0.2 g PVA1788, 0.3 g MC-400, 1.5 g drug, and 0.5 g glycerol). The film-forming gel maintained relatively good appearance. When the amount of anhydrous ethanol added was 1.5 g, its appearance and mechanical properties were even better. Therefore, the amount of anhydrous ethanol added was 1.5 g, as shown in Table 7.

[0080] Table 7 Screening of the amount of anhydrous ethanol used for film-forming gel

[0081]

[0082] 4.8 Drug dosage screening

[0083] The amount of drug added was investigated, and different amounts of drug, 0.1g, 0.3g, 0.5g, 0.7g, and 0.9g, were added to the formula (0.2g PVA, 0.3g MC-400, 0.5g propylene glycol, and 1.5g anhydrous ethanol). When the amount of drug added exceeded 0.7g, cracks appeared in the film after the film-forming gel was formed. When the amount of drug added was 0.5g, the appearance and corresponding mechanical properties of the film-forming gel were relatively good. Therefore, 0.5g was selected as the amount of drug added, as shown in Table 8.

[0084] Table 8 Screening of film-forming gel solution dosage

[0085]

[0086]

[0087] 4.9 Orthogonal experiment

[0088] Based on the results of the single-factor experiments, three factors were screened for their significant influence on the formation of the Qufengchubi film-forming gel for subsequent orthogonal experiments. These factors were film-forming agent dosage (A), plasticizer dosage (B), and ethanol dosage (C), as shown in Table 9. A comprehensive score of the Qufengchubi film-forming gel's performance was used as an evaluation metric. The film-forming time (t) and appearance of the film-forming gel affect the user experience. Excessive film-forming time or uneven film formation can lead to poor user compliance. An ideal film-forming gel should have an appropriate film-forming time, good film-forming properties, and form a uniform film at the point of use with good flexibility and ease of removal. Therefore, a comprehensive score was determined using film-forming time and appearance as indicators. The comprehensive score = (15 - t) + (film-forming properties + flexibility + removability). The Qufengchubi film-forming gel formulation was optimized and analyzed, and the final formulation was determined based on the comprehensive score. The results of the orthogonal experimental design indicate that the influence of different factors on the experimental results follows the order A > C > B. This indicates that the film-forming agent (PVA1788) has the greatest impact, followed by anhydrous ethanol, and the plasticizer (glycerol) has the least. Combining the orthogonal experimental design and the results of analysis of variance, the optimal preparation process for the Qufengchubi film-forming gel was determined to be A1B2C1 (see Tables 10 and 11).

[0089] Table 9 Orthogonal experiment investigation table

[0090]

[0091] Table 10 Orthogonal experiment screening of the formula of the film-forming gel for dispelling wind and removing numbness

[0092]

[0093]

[0094] Table 11 Results of variance analysis of orthogonal experiment

[0095]

[0096] 4.10 Verification test

[0097] To verify the feasibility of the preferred solution, three batches of samples were selected for testing based on the optimal preparation process conditions determined by orthogonal experiments, and the scores for each batch were calculated. The scores for the three batches were 33.24, 34.26, and 33.57, respectively, with an average of 34.03 and an RSD of 1.17%. Therefore, this preparation process is relatively reliable and can be used as the optimal preparation process. The formula is shown in Table 12.

[0098] Table 12 Prescription of Qufengchubi Film-forming Gel

[0099]

[0100] The preparation method is as follows: After weighing 0.3g MC-400 and 0.1g PVA1788, add 0.5g glycerol to moisten the matrix, add 4g deionized water, let it swell at room temperature for 12h, add 0.5g drug, add 1g anhydrous ethanol, stir evenly, and add the remaining water to prepare a total of 10g of film-forming gel.

[0101] The efficacy and safety of the wind-removing and arthralgia-removing film-forming gel obtained under the optimal preparation process were verified.

[0102] Example 2 Verification of the efficacy of the film-forming gel for dispelling wind and removing numbness

[0103] 1 Analgesia experiment

[0104] Forty KM mice were randomly divided into five groups, half male and half female, with eight mice in each group. The groups were: a control group, a Voltaren-positive drug group, and high-, medium-, and low-dose groups of Qufengchubi film-forming gel. The high-, medium-, and low-dose Qufengchubi film-forming gel doses were 0.125g / kg, 0.075g / kg, and 0.025g / kg, respectively. The Voltaren-positive drug group received 0.05g / kg. Drug administration began after seven days of adaptive feeding. One day before administration, the mice's backs were depilated using an animal depilator, covering an area of ​​2cm×3cm. Each drug group was then applied to the depilated area on the backs of the mice. A blank gel was applied to the backs of the control group. Drug administration continued for five consecutive days. One hour after the last dose, each mouse was intraperitoneally injected with 0.9% glacial acetic acid at a dose of 10mL / kg. The number of writhing times within 15 minutes was observed, and the inhibition rate was calculated using the formula. The standard for writhing is that the mouse's abdomen is sunken, the body is elongated, and the buttocks are raised. If all three occur, it is considered a writhing.

[0105] Writhing inhibition rate = (average number of writhing times in the model group - average number of writhing times in the drug-treated group) / average number of writhing times in the model group × 100%

[0106] The experimental results showed that compared with the model group, the number of writhing reactions in the high- and medium-dose Qufengchubi film-forming gels were significantly different, and the differences were statistically significant (P<0.001). Among them, the pain inhibition rate in the high-dose Qufengchubi group was greater than 50%, and it had a certain analgesic effect, as shown in Table 13.

[0107] Table 13 Analgesia experiment (x±s, n=8)

[0108]

[0109] Note: Compared with the blank group, "***" indicates P < 0.001.

[0110] 2 Rheumatoid arthritis experiments

[0111] 2.1 Modeling and drug administration

[0112] Thirty-six male SD rats were randomly divided into six groups, with six rats in each group. After one week of adaptive feeding, a rheumatoid arthritis model was established. 0.1 mL of complete Freund's adjuvant was injected into the right hind paw of the rats, except for the blank group, which received an equal volume of normal saline.

[0113] Rats with successfully established models were randomly divided into three groups: a blank group, a control group, a Voltaren-positive group, and high-, medium-, and low-dose Qufengchubi film-forming gel groups. Following grouping, the corresponding medication was administered daily. The high, medium, and low doses of Qufengchubi film-forming gel were 0.25 g / kg, 0.15 g / kg, and 0.05 g / kg, respectively. Daily dosing began on day 14 after model establishment. The Voltaren-positive group received 0.25 g / kg. Rats in the blank and model groups received a drug-free blank matrix.

[0114] 2.2 Observation indicators

[0115] (1) Swelling of rat toes

[0116] From the day before modeling to the end of drug administration, the swelling degree of the rat's right hind paw was measured every four days using a tissue swelling meter. The swelling degree of the rat's paw was calculated.

[0117] (2) Rheumatoid arthritis score in rats

[0118] From the day before modeling to the end of dosing, the rat arthritis score was recorded every four days using a scoring system, with the maximum score being 16. A score of 0 indicates no redness or swelling; a score of 1 indicates swelling or redness in one to two interdigital joints; a score of 2 indicates swelling or redness in three to four interdigital joints or one larger joint; a score of 3 indicates swelling or redness in more than four joints; and a score of 4 indicates severe arthritis in the entire paw.

[0119] (3) Determination of the content of rat inflammatory factors TNF-α, IL-6 and IL-1β

[0120] After the administration, the rats were fasted and anesthetized with intraperitoneal injection of sodium pentobarbital. Blood was collected from the abdominal aorta, kept at room temperature for 2 h, centrifuged at 3000 r / min for 10 min, and the supernatant was stored in a -80°C refrigerator and tested according to the instructions of the ELISA kit.

[0121] (4) Observation of pathological sections of rat swollen toes

[0122] The pathological changes of ankle joints of AIA rats were observed under a microscope by hematoxylin-eosin (HE) staining.

[0123] 3 Rheumatoid Arthritis Experiment Results

[0124] 3.1 Measurement of rat toe thickness

[0125] After modeling, the paw swelling of the model group and each drug-treated group was significantly increased compared with the blank group (P<0.05), and there was no difference between the model group and each drug-treated group, indicating that the modeling was successful. After treatment, on the 32nd day, the paw swelling of the positive drug group and each drug-treated group was significantly reduced compared with the model group (P<0.05), indicating that the Qufengchubi film-forming gel has a significant inhibitory effect on the paw swelling of rats. Figure 1 .

[0126] 3.2 Rheumatoid arthritis score in rats

[0127] After modeling, the model group showed significant differences compared with the blank group (P<0.001). After drug administration, the arthritis index of rats in each drug administration group decreased compared with the model group. On the 32nd day, compared with the model group, the foot swelling of the positive drug group and each drug administration group was significantly reduced (P<0.05). Figure 2 .

[0128] 3.3 Determination of TNF-α, IL-6, and IL-1β Contents in Rats

[0129] Compared with the blank group, the levels of TNF-α, IL-6 and IL-1β in the serum of the model group rats were significantly increased (P<0.001); compared with the model group, the levels of TNF-α, IL-6 and IL-1β in the high and medium dose groups were significantly decreased (P<0.01), indicating that the Qufengchubi film-forming gel has a certain anti-inflammatory effect. Figure 3 .

[0130] 3.4 Observation of histopathological sections of swollen toes of rats

[0131] Figure 4The results showed that the ankle cartilage surface of rats in the normal group was smooth, with no cartilage damage or inflammatory cell infiltration observed. However, the ankle joints of rats in the model group showed extensive inflammatory cell infiltration. Compared with the model group, the medium- and high-dose Qufengchubi film-forming gel groups showed less pronounced pathological effects and proliferation. Therefore, the medium- and high-dose Qufengchubi film-forming gel groups showed significant anti-inflammatory effects.

[0132] Example 3 Safety Verification of the Film-forming Gel for Expelling Wind and Removing Bi

[0133] Skin irritation test

[0134] Eight healthy rabbits weighing 2.0-2.5 kg (half male and half female) were selected and randomly divided into groups after a three-day pre-feeding period. Twenty-four hours before the experiment, the skin on both sides of the rabbits' backs was shaved. An area free of erythema, lesions, or scabs was selected, and a 2.5 cm × 2.5 cm area was marked with a marker as the administration site. Preparation of damaged skin: Before administration of the test substance, a sterile needle was used to score a "well" on the shaved area of ​​the damaged skin group. The cuticle was marked when bleeding occurred. Using the same left-right self-control method, a dose of 0.5 g / rabbit was applied to the left side of the damaged skin and the right side of the intact skin groups. The patch was applied for at least 4 hours after administration. After application, the test substance was removed and the administration site was cleaned with warm water. Dosing was done once daily for 7 consecutive days.

[0135] Skin reactions were observed visually under natural light. Observe and record any pigmentation, hemorrhages, rough skin, or thin skin at the application site, along with their onset and resolution times, 1 hour after each medication removal and before reapplication, as well as 1 hour, 24 hours, 48 ​​hours, and 72 hours after discontinuation. Erythema and edema were evaluated.

[0136] The results showed that during the experimental period, no erythema or edema, or significant bleeding spots, were observed at the site of application of the Qufengchubi film-forming gel or blank matrix in either the intact or damaged skin groups. No abnormalities, such as erythema or edema, were observed during the withdrawal observation period 1, 24, 48, or 72 hours after the final dose. Based on evaluation criteria such as the skin irritation response score and skin irritation intensity standard, the cumulative skin irritation index for both the intact and damaged skin groups was 0, indicating a non-irritating skin irritation intensity (see Table 14).

[0137] Table 14 New Zealand rabbit multiple dose skin irritation results

[0138]

[0139] References

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[0164] Although the present invention has been described in detail above using general descriptions and specific embodiments, it will be apparent to those skilled in the art that modifications and improvements may be made thereto. Therefore, such modifications and improvements, without departing from the spirit of the present invention, are intended to be within the scope of protection claimed herein.

Claims

1. A film-forming gel for dispelling wind and removing numbness, characterized in that: The anti-wind and anti-bi film-forming gel is 10 parts by weight, comprising: 0.1-0.9 parts of drugs, 0.1-0.5 parts of MC-400, 0.1-0.5 parts of PVA1788, 0.1-0.9 parts of glycerol, 1-3 parts of anhydrous ethanol, and the balance being distilled water. The drugs are prepared using the stems and leaves of Aconitum, Ephedra, Astragalus, White Peony Root and Radix Glycyrrhizae as raw materials.

2. The film-forming gel for dispelling wind and removing numbness according to claim 1, characterized in that include: 0.5 parts of drug, 0.3 parts of MC-400, 0.1 parts of PVA1788, 0.5 parts of glycerol, 1 part of anhydrous ethanol, and the balance is distilled water.

3. The film-forming gel for dispelling wind and removing numbness according to claim 1, characterized in that The preparation method of the medicine is as follows: The crude powder of Aconitum stem and leaf was added into the liquid at a ratio of 20 times the amount of 0.1 mol·L -1 After soaking in hydrochloric acid solution for 1 hour, the mixture was extracted at 70°C for 3 hours, twice, and the extract was concentrated and adjusted to pH 3 using hydrochloric acid. The extract was then loaded onto a 001×12 type cation exchange resin for enrichment and purification, thereby obtaining the total alkaloids from the stems and leaves of Aconitum carmichaelii. Separately, four herbs, namely, ephedra, astragalus, white peony root, and glycyrrhiza uralensis, were mixed in a mass ratio of 1:1:1:

1. Six times the amount of the mixed herbs was added to 50% ethanol and soaked for 30 minutes. The mixture was then condensed and refluxed for three times. The extract was concentrated under vacuum at 60°C to a concentration of 0.2 g of the raw herbs per mL. The total alkaloids from the stems and leaves of Aconitum carmichaelii were mixed with the extracts of the other four medicinal herbs at a ratio of 1:3 and concentrated to a crude drug amount of 15 g·mL. -1 The obtained medicine is stored in a -4°C refrigerator.

4. The method for preparing the film-forming gel for dispelling wind and eliminating numbness according to any one of claims 1 to 3, characterized in that: The method comprises: weighing MC-400, PVA1788 and glycerol according to the prescribed amount, adding part of distilled water to swell and stir overnight, adding the prescribed amount of anhydrous ethanol and the drug to the gel solution in sequence while stirring, stirring to disperse them evenly, and finally adding the remaining distilled water and mixing them evenly.

5. The method for preparing the film-forming gel for dispelling wind and removing numbness according to claim 4, wherein: The mass of the partial distilled water accounts for 50-60% of the total mass of the distilled water.

6. Use of the Qufengchubi film-forming gel according to claim 1 in the preparation of medicines for rheumatoid arthritis.

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