Aegicerocarpus compositus cream, its preparation method and application

Abstract

The present application relates to the technical field of medicine, in particular to a kind of Ai Huangcao cream and its preparation method and application.The Ai Huangcao cream formula is composed of Ai Naxiang oil, egg yolk oil, radix ligustici glauci oil, white vaseline, stearic acid, glycerin monostearate, liquid paraffin, propyl gallate, Tween-80, triethanolamine, sodium dodecyl sulfate, glycerol and water.The Ai Huangcao cream prepared by the preparation method of the present application has good appearance, ductility, centrifugal stability, cold resistance and heat resistance stability.Meanwhile, through skin irritation experiment and rat burn and scald experiment, it is confirmed that the Ai Huangcao cream is safe and has no skin irritation, and has good therapeutic effect on rat grade II burn and scald model.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to an Artemisia argyi cream, its preparation method, and its application. Background Technology

[0002] Burns are common traumatic injuries to the skin and subcutaneous tissues. The main causes include high temperature, heat, chemicals, and electric current. The main clinical manifestations are severe pain at the wound site, local redness and swelling, blister formation, and tissue ulceration. If not treated properly, secondary complications such as wound infection, delayed healing, and scar hyperplasia can easily occur, seriously affecting the patient's quality of life.

[0003] Currently, there are numerous topical preparations used clinically for the treatment of burns and scalds. Western medicine preparations (such as silver sulfadiazine and silver nitrate) primarily focus on antibacterial, anti-inflammatory, and local analgesic effects. While they can effectively control wound infection, their role in promoting tissue regeneration and repair and inhibiting scar formation is limited. Furthermore, some of these drugs have drawbacks such as strong local irritation and the potential for inducing drug resistance with long-term use. In contrast, traditional Chinese medicine topical preparations often follow the principles of clearing heat and detoxifying, promoting blood circulation and relieving pain, and promoting wound healing and tissue regeneration, demonstrating unique advantages in the comprehensive treatment of burns and scalds. However, existing Chinese medicine burn ointments generally suffer from complex prescriptions, cumbersome preparation processes, poor wound penetration, long healing periods, and a high risk of scarring, making it difficult to fully meet the clinical demand for highly effective, safe, and convenient treatment drugs.

[0004] Blumeabalsamifera oil is a natural volatile oil extracted from the plant Blumeabalsamifera, belonging to the Asteraceae family. Modern pharmacological studies have confirmed that Blumeabalsamifera oil possesses anti-inflammatory, analgesic, broad-spectrum antibacterial, wound-healing, and local edema-reducing biological activities. It can effectively inhibit the inflammatory response of burn wounds, accelerate epidermal cell regeneration, and shorten the time for scab formation and healing. Egg yolk oil is a fat-soluble component extracted from egg yolks through high-temperature refining. It is rich in nutrients such as vitamin A, vitamin D, and lecithin. It can nourish wounds, repair damaged skin tissue, relieve burning pain, and form a physical protective film on the wound surface, reducing external irritants and pathogenic microorganisms, thereby promoting the healing of burn wounds. Lithospermum erythrorhizon oil is a topical oil preparation made by extracting lithospermum erythrorhizon through oil extraction. Its core functions are cooling blood and detoxifying, promoting tissue regeneration, clearing heat and relieving pain. It can quickly relieve redness and pain in burn wounds, inhibit the proliferation of pathogenic bacteria in the wound, accelerate the repair process of ulcerated tissue, and reduce the formation of scar tissue.

[0005] Currently, there are no reports in existing technologies of scientifically combining senna oil, egg yolk oil, and comfrey oil to prepare a topical cream for burn treatment. Using any one of these oil components alone has limitations in its efficacy: senna oil excels in anti-inflammatory and analgesic effects but lacks nutritional repair and film-forming protection; egg yolk oil focuses on nourishing the wound and providing physical isolation, with relatively insufficient anti-inflammatory and antibacterial capabilities; comfrey oil excels at cooling the blood and promoting tissue regeneration, but its effects are relatively slow. A reasonable combination of these three components is expected to achieve complementary functions and synergistic effects: senna oil can play a pioneering role in anti-inflammatory and analgesic effects; egg yolk oil provides logistical support for nutritional repair and physical protection; and comfrey oil acts as a "cleaner" for cooling the blood, antibacterial properties, and promoting tissue regeneration, thus comprehensively covering the multi-stage and multi-target treatment needs in burn treatment, including anti-inflammatory, analgesic, antibacterial, healing-promoting, and scar-preventing effects. Furthermore, preparing the above-mentioned compound active ingredients into an ointment formulation can improve the physical and chemical stability of the preparation, improve wound adhesion and drug permeability, and facilitate clinical use and storage.

[0006] Based on this, the present invention provides an Artemisia annua cream, the active ingredients of which are composed of Artemisia argyi oil, egg yolk oil, and comfrey oil, and are prepared with a cream base. The present invention aims to solve the above-mentioned technical problems existing in current topical burn preparations, and to provide a burn treatment drug with definite efficacy, high safety, and convenient use for clinical use. Summary of the Invention

[0007] The purpose of this invention is to provide an Artemisia argyi cream and its preparation method.

[0008] Another object of the present invention is to provide the application of the above-mentioned Artemisia argyi cream in the preparation of a drug for promoting the repair of Grade II burn wounds.

[0009] To achieve the above objectives, the technical solution adopted by the present invention is as follows: The formulation of the Artemisia argyi cream of the present invention consists of the following components in weight percentage: 3%-20% of the drug, 5% white petrolatum, 5% stearic acid, 3%-7% glyceryl monostearate, 10% liquid paraffin, 0.2% ethylparaben, 2%-6% Tween-80, 0-1% triethanolamine, 0.5%-1% sodium lauryl sulfate, 8%-12% glycerin, and the balance being water; wherein the drug is composed of Artemisia argyi oil, egg yolk oil, and comfrey oil in a weight ratio of 1-5:1-4:1-5. The egg yolk oil is prepared by the following method: boil eggs, remove egg whites, grind egg yolks into powder, place them in a stainless steel pot, heat over low heat until at least a small amount of oil is produced, stir-fry over high heat until the egg yolks turn brown, filter, remove the residue, and the oil is obtained. The comfrey oil is prepared by the following method: weigh rapeseed oil with a mass of 6 times that of comfrey, heat it to boiling, add comfrey, extract for 5 minutes, filter, and the oil is obtained.

[0010] Preferably, the formulation of the Artemisia argyi cream of the present invention consists of the following components in weight percentage: 9% of the drug, 5% white petrolatum, 5% stearic acid, 5% glyceryl monostearate, 10% liquid paraffin, 0.2% ethylparaben, 4% Tween-80, 0.5% triethanolamine, 0.5% sodium lauryl sulfate, 10% glycerin, and the balance being water; wherein the drug is composed of Artemisia argyi oil, egg yolk oil and comfrey oil in a weight ratio of 1:1:1.

[0011] The preparation method of the Artemisia argyi cream of the present invention is as follows: accurately weigh each component according to the formula; add Artemisia argyi oil, egg yolk oil, comfrey oil, white petrolatum, stearic acid, glyceryl monostearate, liquid paraffin and ethylparaben to the oil phase beaker, and heat in an 80°C water bath until completely dissolved; add glycerin, Tween-80, triethanolamine, sodium lauryl sulfate and water to the aqueous phase beaker, and heat in an 80°C water bath until completely dissolved; slowly add the aqueous phase to the oil phase, emulsify at 300-700 rpm and 60°C-80°C for 5-15 minutes, continuously stir and cool to room temperature to obtain Artemisia argyi cream.

[0012] Preferably, in the preparation method of the Artemisia argyi cream of the present invention, the stirring speed of the emulsification is 500-700 rpm.

[0013] In a further preferred embodiment, in the preparation method of the Artemisia argyi cream of the present invention, the stirring speed of the emulsification is 500 rpm.

[0014] Preferably, in the preparation method of the Artemisia argyi cream of the present invention, the emulsification temperature is 70℃-80℃.

[0015] In a further preferred embodiment, the emulsification temperature in the preparation method of the Artemisia argyi cream of the present invention is 80°C.

[0016] Preferably, in the preparation method of the Artemisia argyi cream of the present invention, the emulsification time is 5-10 min.

[0017] In a further preferred embodiment, the emulsification time in the preparation method of the Artemisia argyi cream of the present invention is 5 minutes.

[0018] The application of the Artemisia argyi cream described in this invention in the preparation of drugs that promote the repair of Grade II burn wounds.

[0019] The beneficial effects of this invention are: 1. This invention provides an Artemisia annua cream with Artemisia argyi oil, egg yolk oil, and comfrey oil as pharmaceutical ingredients. To obtain the optimal formulation and preparation process of this cream, the inventors conducted a series of condition investigation experiments: Through single-factor experiments, the optimal oil loading (9%, Artemisia argyi oil: egg yolk oil: comfrey oil = 1:1:1), the optimal emulsifier and its dosage (4% Tween-80, 0.5% sodium lauryl sulfate, and 0.5% triethanolamine) were screened out. At the same time, the dosage of glyceryl monostearate was identified as the factor that has the greatest impact on the cream's molding. Through response surface methodology, the three main factors affecting the molding effect and their optimal dosages (5% glyceryl monostearate, 10% glycerin, and 4% Tween-80) were obtained. Through orthogonal experiment optimization, the optimal preparation process conditions of this cream were determined (stirring speed 500 rpm, emulsification temperature 80℃, emulsification time 5 min, and emulsification method of adding the oil phase to the aqueous phase). Under the above-mentioned optimal formulation and process conditions, the prepared cream formulation exhibits good appearance, spreadability, centrifugal stability, cold resistance, and heat resistance.

[0020] 2. This invention investigated the safety of the cream and its therapeutic effect on grade II burns through mouse skin irritation experiments and rat burn experiments. Results showed that the *Artemisia argyi* cream provided by this invention is safe and non-irritating to the skin; pathological section results showed that the *Artemisia argyi* cream effectively promoted the repair of grade II burn wounds and reduced scar formation; ELISA results showed that normal doses of *Artemisia argyi* cream significantly downregulated TNF-α and IL-6 levels and upregulated IL-10 levels, bringing them back to the levels in the control group, thus regulating the inflammatory response. These results indicate that the *Artemisia argyi* cream provided by this invention has a good therapeutic effect on a rat model of grade II burns, and can regulate immunity and accelerate burn recovery by controlling the levels of anti-inflammatory / pro-inflammatory markers TNF-α, IL-6, and IL-10. Attached Figure Description

[0021] Figure 1 The results of response surface methodology were used to optimize the formulation. Figure 2 These are the results of a skin irritation test. Figure 3 The results of wound healing in each group of rats were statistically analyzed. Figure 4 HE staining image of rat burn tissue (scale bar 200 μm); Figure 5 The changes in anti-inflammatory / pro-inflammatory marker levels in each group of animals. Detailed Implementation

[0022] The technical solution of the present invention will be described in detail below with reference to specific embodiments. The following embodiments are for explanation and illustration only, and do not constitute a limitation on the technical solution of the present invention.

[0023] Example 1

[0024] Artemisia argyi cream, formula (200 g): 6 g Artemisia argyi oil, 6 g egg yolk oil, 6 g comfrey oil, 10 g white petrolatum, 10 g stearic acid, 10 g glyceryl monostearate, 20 g liquid paraffin, 20 g glycerin, 8 g Tween-80, 1 g triethanolamine, 1 g sodium lauryl sulfate, 0.4 g ethylparaben, 101.6 g water.

[0025] Example 2

[0026] Artemisia argyi cream, formula (200 g): 2 g Artemisia argyi oil, 2 g egg yolk oil, 2 g comfrey oil, 10 g white petrolatum, 10 g stearic acid, 6 g glyceryl monostearate, 20 g liquid paraffin, 16 g glycerin, 4 g Tween-80, 1 g sodium lauryl sulfate, 0.4 g ethylparaben, 126.6 g water.

[0027] Example 3

[0028] Artemisia argyi cream, formula (300 g): 20 g Artemisia argyi oil, 20 g egg yolk oil, 20 g comfrey oil, 15 g white petrolatum, 15 g stearic acid, 21 g glyceryl monostearate, 30 g liquid paraffin, 36 g glycerin, 18 g Tween-80, 3 g triethanolamine, 3 g sodium lauryl sulfate, 0.6 g ethylparaben, and 98.4 g water.

[0029] Example 4

[0030] Artemisia argyi cream was prepared using the formulations of Examples 1-3, and the specific steps are as follows: Accurately weigh each component according to the formula; add senna oil, egg yolk oil, comfrey oil, white petrolatum, stearic acid, glyceryl monostearate, liquid paraffin, and ethylparaben to the oil phase beaker and heat in an 80°C water bath until completely dissolved; add glycerin, Tween-80, triethanolamine, sodium lauryl sulfate, and water to the aqueous phase beaker and heat in an 80°C water bath until completely dissolved; slowly add the aqueous phase to the oil phase and emulsify at 500 rpm and 80°C for 5 minutes, stirring continuously and cooling to room temperature to obtain Artemisia argyi cream.

[0031] Example 5

[0032] Artemisia argyi cream was prepared using the formulations of Examples 1-3, and the specific steps are as follows: Accurately weigh each component according to the formula; add senna oil, egg yolk oil, comfrey oil, white petrolatum, stearic acid, glyceryl monostearate, liquid paraffin, and ethylparaben to the oil phase beaker and heat in an 80°C water bath until completely dissolved; add glycerin, Tween-80, triethanolamine, sodium lauryl sulfate, and water to the aqueous phase beaker and heat in an 80°C water bath until completely dissolved; slowly add the aqueous phase to the oil phase and emulsify at 300 rpm and 60°C for 15 min, stirring continuously and cooling to room temperature to obtain senna cream.

[0033] Example 6

[0034] Artemisia argyi cream was prepared using the formulations of Examples 1-3, and the specific steps are as follows: Accurately weigh each component according to the formula; add senna oil, egg yolk oil, comfrey oil, white petrolatum, stearic acid, glyceryl monostearate, liquid paraffin, and ethylparaben to the oil phase beaker and heat in an 80°C water bath until completely dissolved; add glycerin, Tween-80, triethanolamine, sodium lauryl sulfate, and water to the aqueous phase beaker and heat in an 80°C water bath until completely dissolved; slowly add the aqueous phase to the oil phase and emulsify at 700 rpm and 80°C for 5 minutes, stirring continuously and cooling to room temperature to obtain senna cream.

[0035] To further verify the reliability of the present invention and select the optimal solution, the inventors conducted a series of experiments, as follows: I. Research on the Preparation Process of Artemisia argyi Cream 1. Instruments and reagents The experimental instruments are shown in Table 1, and the drug information is shown in Table 2.

[0036] ; .

[0037] Preparation of egg yolk oil: Boil eggs, remove egg whites, grind egg yolks into powder, place in a stainless steel pot, heat over low heat until at least a small amount of oil is produced, stir-fry over high heat until the egg yolks turn brown, filter, remove residue, and you have the oil.

[0038] Preparation of Lithospermum erythrorhizon oil: Weigh 6 times the weight of Lithospermum erythrorhizon into rapeseed oil, heat to boiling, add Lithospermum erythrorhizon, extract for 5 minutes, filter, and the oil is obtained.

[0039] 2. Preparation process research 2.1 Prescription composition and implementation standards .

[0040] 2.2 Preparation process and evaluation indicators of cream 2.2.1 Preparation process The drug was added to the oil phase beaker and heated in an 80°C water bath until completely dissolved. The emulsifier was added to the aqueous phase beaker and heated in an 80°C water bath until completely dissolved. After dissolution, the aqueous phase was slowly added to the oil phase, and the mixture was stirred at 800 rpm using a magnetic stirrer until it reached the desired consistency. This experiment used appearance, spreadability, centrifugal stability, cold resistance, and heat resistance as evaluation indicators to investigate the process of the Artemisia argyi cream.

[0041] 2.2.2 Evaluation Indicators (1) Appearance: Observe the state of the creams with different formulations, apply them to the skin, and then clean them. The creams should be uniform and delicate, without roughness, of moderate thickness, easy to apply, and easy to wash off. Analyze and score them.

[0042] (2) Spreadability: Observe the state of creams with different formulations, apply them to a glass plate, and then spread them. The cream should be uniform and smooth, without roughness, of moderate thickness, and easy to spread. Analyze and score the results.

[0043] (3) Centrifugal stability: Take 0.5 g of cream with different formulations and place them in 1.5 mL EP tubes. Then place them symmetrically in a high-speed centrifuge. Set the speed to 7000 r / min and the centrifugation time to 10 min. Then take out the samples for observation, analysis and scoring.

[0044] (4) Cold resistance stability: Take appropriate amounts of creams with different formulations, place them in 5 mL EP tubes, and divide them into two groups. One group is placed in a refrigerator and left to stand at -20℃ for 24 h.

[0045] (5) Heat resistance stability: Take appropriate amounts of creams with different formulations, place them in an oven and heat them to 40℃, 50℃ and 60℃ for 1 hour, then take them out for observation, analysis and scoring.

[0046] The specific scoring criteria are shown in Table 4.

[0047] .

[0048] 2.2.3 Prescription Selection The main components of the cream are the oil phase, aqueous phase, and emulsifier. Four oil phases (white petrolatum, liquid paraffin, stearic acid, and glyceryl monostearate) and emulsifiers (Tween 80 and sodium lauryl sulfate) were screened. The amount of emulsifier in the formulation was determined based on the hydrophilic-lipophilic balance (HLB) value, as shown in the following formula, where A% and B% represent the weight ratio (w / w) of emulsifier and co-emulsifier: ; The preliminary formulation composition obtained from HLB calculations is shown in Table 5. Based on relevant evaluation indicators for creams, the appearance, centrifugal stability, and cold and heat resistance of different formulations were scored to select the optimal formulation.

[0049] .

[0050] 2.3 Single-factor analysis 2.3.1 Selection of oil load A single-factor study was conducted on the oil loading capacity and its proportion. The optimal dosage range was determined by scoring the cream's quality. The dosage and results are shown in Table 6, with the base dosage as shown in Table 5.

[0051] ; The results show that the cream with the highest score has the best molding quality when the oil loading is 9%. Therefore, the oil loading in the formula should be 9%.

[0052] 2.3.2 Optimal ratio of sage oil, egg yolk oil, and comfrey oil Based on the 9% oil loading selected from the single-factor experiments above, the proportions of each component were screened and configured as shown in Table 7. The optimal ratio was determined by examining its properties, and the results are shown in Table 7.

[0053] ; The results showed that the cream formed better when the oil ratio was "Artemisia:Egg:Purple = 1:1:1", so the oil ratio was determined to be 1:1:1.

[0054] 2.3.3 Optimization of Artemisia argyi ointment formulation and preparation process Creams are mainly divided into two types: W / O and O / W. The composition of a cream mainly includes active ingredients (drugs), oil phase matrix, aqueous phase matrix, and a small amount of additives (such as preservatives). The selection and ratio of oil phase matrix and aqueous phase matrix are extremely important for the formation of the entire cream. Moreover, the matrix is ​​also the carrier of active ingredients (drugs). Different matrices will lead to different degrees of absorption of inflammatory exudate.

[0055] Based on the characteristics of creams and the differences in the properties of each base component, this study designed four base formulations in two categories: W / O type cream base and O / W type cream base. The dosage and scoring are shown in Table 8.

[0056] ; The results showed that prescription No. 4 had the highest overall score of 39. Therefore, the optimal emulsifier and its dosage were determined to be: Tween-80 (4%), sodium dodecyl sulfate (0.5%), and triethanolamine (0.5%).

[0057] 2.3.4 Cream formulation and screening of key factors Based on the aforementioned oil loading and its proportion, and the results of emulsifier screening, the formulation of the cream was optimized. Six formulations were designed, and the scores were obtained according to the scoring criteria, as shown in Table 9.

[0058] ; The results showed that prescription No. 1 had the highest overall score of 43. The above experiments revealed that the oil phase with the greatest influence on cream formation was glyceryl monostearate.

[0059] 2.4 Box-Behnken Response Surface Experimental Design and Results 2.4.1 Box-Behnken Response Surface Design Using Box-Behnken Design (BBD) response surface methodology, after determining the excipient types based on the aforementioned stability studies and single-factor experiments, key influencing factors—glyceryl monostearate (A), glycerin (B), and Tween-80 (C)—were selected using Design Expert software V13.0.1.0 to analyze their effects on the appearance and stability of the cream and determine their dosage. Box-Behnken designs were conducted at levels of -1, 0, and 1. Factor levels are shown in Table 10, and the results are shown in Table 11.

[0060] ; .

[0061] 2.4.2 Analysis of Response Surface Methodology Test Results Response surface methodology was performed on the ratings using Design Expert 13 software. The resulting bivariate regression equation for the ratings was Y = -65.3063 + 14.5A + 9.275B + 10.25C - 0.0625AB + 0.125AC + 0.25BC - 1.51875A 2 -1.39375B 2 -0.58125C 2 The analysis of variance (Table 12) shows that the p-value of the experimental model is <0.0001, indicating extremely strong significance. The p-value for the lack-of-fit term is 0.7510 > 0.05, indicating no significance. This suggests that the unknown factors have minimal interference with the model, and the equation has a good fit. The correlation coefficient R0 of the regression equation is... 2 =0.9739, correction factor R 2 =0.9404, indicating that the model's predictions are accurate and can be used for data analysis. Factors A, B, and C had no significant impact on the score (P>0.05); the interaction terms AB, AC, and BC also had no significant impact on the score (P>0.05); the quadratic term A...2 B 2 The impact on the score was extremely significant (P < 0.0001). By comparing the F-values, the influence of the three factors—glyceryl monostearate, Tween-80, and glycerin—on the score was determined, with F-values ​​of 2.31, 2.31, and 1.48, respectively. Therefore, the overall influence of each factor on the molding process of Artemisia argyi cream is in the order of glyceryl monostearate ≈ Tween-80 > glycerin, and none of the three factors reached a significant level (P > 0.05).

[0062] ; See response surface methodology diagram. Figure 1 Observe the shape of the contour plot and the steepness of the response surface. The more elliptical the shape of the contour lines and the steeper the change in the three-dimensional surface, the stronger the interaction between the two factors and the more significant their impact on the score. Figure 1 It can be seen that, although the contour lines of the three groups—glyceryl monostearate and Tween-80, glyceryl monostearate and glycerol, and Tween-80 and glycerol—are slightly elliptical, their principal axes are basically aligned, with low ellipticity and no crossover or twisting, indicating weak interaction between them (P>0.05). This is consistent with the statistical conclusion in Table 12 that the interaction terms AB, AC, and BC are all insignificant (P>0.05). The response surface slope is significantly steep, especially in the high-level region of glyceryl monostearate, where the score changes drastically. Even small fluctuations in dosage can cause large changes in the response value, meaning that the score mutations are mainly caused by the quadratic terms of each factor (A). 2 B 2 The results were highly significant (P < 0.0001), indicating that the factors were dominant rather than synergistic or antagonistic. These results are consistent with those from the analysis of variance.

[0063] 2.4.3 Response Surface Methodology Optimization and Validation Data analysis using Design-Expert revealed the optimal dosage as follows: glyceryl monostearate (5%), glycerol (10%), and Tween-80 (4%), with a predicted score of 43.4. Based on these conditions, a three-parallel validation experiment was conducted, yielding an average comprehensive score of 43, which is close to the predicted value. This indicates that the model's prediction is accurate, and the process is reasonable and feasible.

[0064] 2.5 Orthogonal Experiment to Optimize the Molding Process of Artemisia argyi Cream 2.5.1 Orthogonal Experimental Factor Level Design The dosage forms optimized in the preliminary experiments were screened for formulation using orthogonal experiments, following the L9(3) algorithm. 4An orthogonal array was used to design the experiment. Four factors were considered: stirring speed, emulsification temperature, emulsification time, and emulsification method. Each factor was examined at three levels, as shown in Table 13. An orthogonal experiment was conducted according to the design scheme. The appearance and stability of the selected ointments were examined and scored. The results are shown in Table 14.

[0065] ; ; Based on the above analysis, the optimal process parameters for this experiment are: stirring speed of 500 rpm, emulsification temperature of 80℃, emulsification time of 5 min, and emulsification method of adding the aqueous phase to the oil phase.

[0066] 2.5.2 Validation of Optimal Formulation Process The optimal formulation and preparation process obtained above were validated in three batches in parallel. The dosage of the formulation is shown in Table 15.

[0067]

[0068] The results showed that the average comprehensive score of the three batches was 44, indicating that the dosage and preparation process were optimal. The resulting cream had a fine and smooth texture, uniform color, good spreadability, cold and heat resistance, and excellent stability after centrifugation. This demonstrates that the process has good repeatability and that the process conditions are reliable and stable.

[0069] 3. Summary This chapter, based on single-factor experiments, determined that the oil loading in the Artemisia argyi cream should be 9% (Artemisia:Huang:Cao = 1:1:1), and the optimal emulsifiers and their amounts are: Tween-80 (4%), sodium dodecyl sulfate (0.5%), and triethanolamine (0.5%). Simultaneously, the most significant factor affecting cream formation was identified as glyceryl monostearate in the oil phase. Response surface methodology was then used, and Design-Expert data analysis was employed to determine the amounts of the three main factors influencing the forming effect: glyceryl monostearate (5%), glycerin (10%), and Tween-80 (4%). After determining the matrix amounts, orthogonal experimental design was used to optimize the process, determining the optimal parameters as follows: stirring speed of 500 rpm, emulsification temperature of 80℃, emulsification time of 5 min, and emulsification method of adding the aqueous phase to the oil phase. Through verification of the optimal formulation process, a cream formulation with good appearance, spreadability, cold and heat resistance, and good stability after centrifugation was finally obtained.

[0070] II. Safety and Pharmacodynamic Studies of Artemisia argyi Cream 1. Instruments and reagents 1.1 Instruments ; 1.2 Experimental Reagents and Chemicals .

[0071] 1.3 Preparation of Artemisia annua cream According to the optimal prescription selected in the previous chapter (6 g of senna oil, 6 g of egg yolk oil, 6 g of comfrey oil, 10 g of white petrolatum, 10 g of stearic acid, 10 g of glyceryl monostearate, 20 g of liquid paraffin, 20 g of glycerin, 8 g of Tween-80, 1 g of triethanolamine, 1 g of sodium lauryl sulfate, 0.4 g of ethylparaben, and 101.6 g of water), each component is accurately weighed. The oil phase components (senna oil, egg yolk oil, comfrey oil, white petrolatum, stearic acid, glyceryl monostearate, liquid paraffin, and ethylparaben) and the aqueous phase components (glycerin, Tween-80, triethanolamine, sodium lauryl sulfate, and water) are heated to 80°C respectively. Then, the aqueous phase is slowly added to the oil phase for emulsification. The mixture is continuously stirred and cooled to room temperature to obtain the Artemisia argyi cream.

[0072] 1.4 Laboratory Animals Sixteen SPF-grade KM mice (half male, half female, weighing 20±2 g) were purchased from the Animal Experiment Institute of Guizhou University of Traditional Chinese Medicine. Animal housing environment: temperature (23±2)℃, humidity (55±5)%, 12-hour light-dark cycle, acclimatization for one week, with free access to standard rodent feed and sterile water. All animal experimental protocols were reviewed and approved by the Experimental Animal Ethics Committee of Guizhou University of Traditional Chinese Medicine (Approval No.: 20250808001). Seventy-two SPF-grade SD rats (half male, half female, weighing 200±20 g) were also purchased from the Animal Experiment Institute of Guizhou University of Traditional Chinese Medicine. Animal housing environment: temperature (23±2)℃, humidity (55±5)%, 12-hour light-dark cycle, acclimatization for one week, with free access to standard rodent feed and sterile water. All animal experimental protocols were reviewed and approved by the Experimental Animal Ethics Committee of Guizhou University of Traditional Chinese Medicine (Approval No.: 20250724001).

[0073] 2. Methods and results of mouse skin irritation test 2.1 Experimental Grouping The experiment was divided into two groups, A and B, with 8 animals in each group. The samples were physiological saline (control) and Artemisia argyi cream, respectively. Each group also included an intact skin group and a damaged skin group.

[0074] 2.2 Experimental Methods Remove approximately 2 cm × 2 cm of hair from the back of the mice using a depilatory agent or scissors. Carefully examine the skin to ensure there is no damage. If damage is found, replace the mouse with another. Grouping and administration: Group A received 100 mg of physiological saline applied to the depilated area; Group B received 100 mg of Artemisia argyi oil cream applied to the depilated area.

[0075] Cleaning and observation: 24 hours after administration, gently wash the test site with warm water to ensure complete removal of all residual drug. Observe the erythema and edema at the application site at 4 h, 24 h, 48 h, and 72 h after removal of residual drug. Score according to the criteria in Table 18, and derive the results from Table 19.

[0076] ; .

[0077] 2.3 Experimental Results Skin irritation test results as follows Figure 2 As shown in the results, during the 72-hour experimental period, no skin reactions or signs of irritation (edema, erythema, crusting) were observed in either the test sites treated with Artemisia argyi oil cream or the control group. The final score was in the range of "0-0.5", and based on the scores in Tables 18 and 19, this category was determined to be "non-irritating" under all test conditions.

[0078] 3. Experimental study on the healing of deep second-degree burns treated with Artemisia argyi cream 3.1 Experimental Grouping Seventy-two healthy SD rats, half male and half female, were randomly divided into six groups: blank group (no treatment), model group (modeling only), positive drug group (Mebo moist burn ointment), blank matrix group (cream without drug), Artemisia argyi cream group, and high-dose Artemisia argyi cream group.

[0079] 3.2 Experimental Methods 3.2.1 Preparation of a rat skin burn induction model Rats were first anesthetized, and the hair on their backs was removed. The skin on their backs was then washed with clean water and dried. A rat burn model was created by applying a metal medium to the skin in contact with the spine. The medium was horizontally applied to the skin over the hairless area on the rat's back to prevent damage to underlying organs and to ensure a uniform, circular burn area. The temperature was set to 100℃, and the application time was 8 seconds to establish a second-degree burn model. After modeling, a prepared ointment was evenly applied to the burn wound area twice daily, morning and evening, at a dose of 0.5 g (1 g for the high-dose group). The model group and the control group received no treatment. The total treatment course was 2 weeks.

[0080] 3.2.2 Rat Experimental Sample Collection On days 7 and 14 after modeling, a group of rats were sacrificed, and venous blood was obtained from the rats via the abdominal aorta. The blood was then centrifuged at 5000 rpm for 15 min, and the supernatant was collected for later use. Subsequently, the skin tissue from the rat wounds was cut off and preserved in a fixative while maintaining a flat appearance.

[0081] 3.2.3 Recording and evaluation of burn wounds After modeling, the wound condition was photographed every three days, and the diameter and outline of the rat wound were measured with a ruler to facilitate subsequent analysis of wound area changes. The wound area was calculated using ImageJ software.

[0082] 3.2.4 Histopathological examination Tissue was fixed with paraffin embedding fixative, and then sectioned into 4 µm sections using a microtome. The sections were stained with hematoxylin and eosin (H&E) and observed under a microscope.

[0083] 3.2.5 Determination of pro-inflammatory / anti-inflammatory biomarkers The levels of pro-inflammatory cytokines IL-6, IL-10, and TNF-α in tissue serum were detected using an ELISA kit.

[0084] 3.3 Experimental Results 3.3.1 Observation of burn wound morphology in rats After modeling, the wound was photographed every three days to record changes in the wound's condition and area. The results are as follows: Figure 3 As shown in the results, after 3 days, necrosis and edema appeared in all groups except the control group, and scab formation and edema were also observed. At this time, the wound area of ​​all groups except the control group was significantly reduced compared with the model group (P < 0.05). After 6 days, all wounds had scabbed over and begun to shrink, and some wound edges had begun to fall off. Compared with the model group, the wound area of ​​the positive group, cream group, and high-dose group was significantly smaller (P < 0.05), while the area of ​​the matrix group was significantly larger (P < 0.05). At 9 days, large-area scab formation or scab fall-off was observed in all wounds. Compared with the model group, except for the control group, the wound area of ​​all groups was significantly smaller (P < 0.05). All external wounds showed significant shrinkage (P < 0.05); at 12 days, except for the blank control group, all scabs had fallen off, but a small amount of blood scabs remained. At this time, except for the blank control group, the wound area of ​​all groups was significantly reduced compared with the model group (P < 0.05); at 14 days, there was no statistically significant difference in wound area between the cream group and the high-dose group and the blank control group (P > 0.05). The wounds of the cream group and the high-dose cream group healed without leaving obvious scars, indicating that Artemisia argyi cream has significant wound healing activity. At this time, the wound area of ​​the matrix group and the positive group was significantly reduced compared with the model group (P < 0.05).

[0085] 3.3.2 Histopathological examination of burns in rats To further observe the growth of new tissue in burn wounds, HE staining was performed on wounds on days 7 and 14, followed by histopathological analysis. The results are shown below. Figure 4The results showed that on day 7, inflammatory cells were observed to accumulate at the wound site in all groups except the control group. In both control groups, the epidermis of the rats was relatively intact, with a smooth junction between the epidermis and dermis; collagen fibers in the dermis were loosely arranged, generally parallel to the epidermis; hair follicles were mostly in the growth phase, running obliquely downwards through the dermis; the subcutaneous structure was intact, muscle bundles were aligned, and no obvious hemorrhage, necrosis, or extensive inflammatory cell infiltration was observed; the skin tissue structure was basically normal. In the model group, epidermal necrosis occurred on day 7, the wound was covered with a thick scab, dermal blood vessels were dilated, fibroblasts and inflammatory cells were mixed, and hair follicles were broken and disappeared. On day 14, only a thin scab and a few layers of newly formed epithelium were observed; collagen began to arrange but remained disordered. In the control matrix group, the scab was thick on day 7, capillary hyperplasia was excessive, and inflammatory cells were numerous and disordered. On day 14, a bare area remained in the center, collagen thickening was limited, and short epithelial cords were observed in the hair follicle remnants. On day 7, the scab in the positive control group was significantly thinner, capillary density was controlled, inflammation was mild, and epithelial cords appeared around the hair follicles. On day 14, the epidermis was completely closed, collagen bundles were mostly parallel to the epidermis, and epithelial cords extended to form cystic swellings, i.e., hair follicle bud formation. On day 7, the normal dose of Artemisia argyi cream had the least exudation, capillaries were comparable to those in the positive control group, inflammatory cells were few and sparse, and epithelial cords appeared earlier than in the positive control group. On day 14, most of the collagen was parallel to the epidermal area, the epithelial cords further elongated, and multiple hair follicle buds were seen. The skin remodeling effect was better than that of the positive control group. On day 7, the high dose of Artemisia argyi cream had a thicker scab, more aggregated capillaries, more and more disordered inflammatory cells, and slower coverage of the micelles and epidermis. Although epidermal closure and partial collagen parallelism were achieved on day 14, hair follicle structures were not observed, suggesting that the increased cream concentration did not improve efficacy and may even have inhibited the regeneration of skin appendages.

[0086] 3.3.3 Levels of anti-inflammatory / pro-inflammatory markers in treated and untreated animals The serum levels of TNF-α, IL-6, and IL-10 in each group of rats were detected by ELISA, and statistical analysis was performed. The results are as follows: Figure 5 .

[0087] The results showed that, compared with the blank control group, the levels of TNF-α and IL-6 in the model group were significantly upregulated (p < 0.05), while the level of IL-10 was significantly downregulated (p < 0.05) at 7 days. After drug intervention, all groups showed some degree of regression compared with the model group.

[0088] IL-6 levels: At day 7, all groups showed a significant upregulation compared to the control group (P < 0.05). All treatment groups showed a significant downregulation compared to the model group (P < 0.05), but still did not return to the control group levels. At day 14, all groups showed a significant upregulation compared to the control group (P < 0.05), with the matrix group, positive control group, and cream group showing significant regression effects compared to the model group (P < 0.05). The high-dose group showed no significant difference between day 14 and the model group, or between day 7 and the high-dose group (P > 0.05), and the cream group showed significantly better efficacy than the high-dose group on day 14 (P < 0.05). IL-6 levels gradually decreased in most groups over time.

[0089] IL-10 levels: On day 7, all groups showed a significant decrease compared to the control group (P < 0.05), while the matrix group, positive control group, cream group, and high-dose group showed a significant increase compared to the model group (P < 0.05). On day 14, all groups except the positive control group showed a significant decrease compared to the control group (P < 0.05), while the matrix group, positive control group, and cream group showed a significant increase compared to the model group (P < 0.05). The high-dose group showed a significant decrease compared to the cream group (P < 0.05), but no significant difference compared to the high-dose group on day 7 (P > 0.05).

[0090] TNF-α levels: On day 7, all groups showed a significant increase compared to the control group (P < 0.05), and all groups except the control group showed a significant decrease compared to the model group (P < 0.05). On day 14, the model group, matrix group, and positive control group showed significant upregulation compared to the control group (P < 0.05), and all groups except the control group showed significant downregulation compared to the model group (P < 0.05). Compared to the 7-day high-dose group, the TNF-α level in the 14-day high-dose group was significantly lower (P < 0.05), and there was no statistically significant difference compared to the cream group on day 14 (P > 0.05).

[0091] 4. Discussion The irritation of Artemisia argyi cream was simulated and verified, and the results showed that it was safe and non-irritating to the skin. Using rats with second-degree burns as a model, a simple pharmacological analysis of Artemisia argyi cream was conducted to assess its macroscopic recovery and efficacy. The results showed that after treatment with Artemisia argyi cream, the macroscopic wound area in the normal dose group was significantly smaller than that in the model group (P < 0.05), and the wound healing speed was significantly faster. After HE staining, the number of newly formed skin appendages after treatment with Artemisia argyi cream was significantly greater than that in the model group (P < 0.05), indicating that the cream group was more effective in promoting wound recovery and reducing scar formation. The corresponding anti-inflammatory / pro-inflammatory factors TNF-α and IL-6 significantly reverted to the control group with prolonged treatment time (P < 0.05), while IL-10 was significantly upregulated (P < 0.05), showing good bidirectional immune regulation. The levels of IL-6 and TNF-α in the matrix group, positive group, and model group all showed a significant decreasing trend (P < 0.05), while the level of IL-10 significantly increased (P < 0.05). This indicates that the positive drug (moist burn ointment) has a significant therapeutic effect on controlling burn inflammation, while the blank matrix may alleviate inflammation due to the moisturizing effect of some matrix components. Notably, there was no significant difference in macroscopic wound area between the two doses of Artemisia argyi cream (normal dose and high dose) (P > 0.05). However, HE staining and ELISA tests revealed that the recovery of skin appendages in the high-dose cream group was slower than that in the normal dose group at the same time point. The levels of IL-6 and IL-10 in the high-dose group did not significantly revert to the levels in the blank group with the extension of treatment time (P > 0.05), and there was a significant difference compared with the cream group at the same time point (P < 0.05). At 14 days, there was no statistically significant difference compared with the model group (P > 0.05).

[0092] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A cream of Artemisia vulgaris, characterized in that, The formulation of the Artemisia argyi cream consists of the following components in weight percentage: 3%-20% of the drug, 5% white petrolatum, 5% stearic acid, 3%-7% glyceryl monostearate, 10% liquid paraffin, 0.2% ethylparaben, 2%-6% Tween-80, 0-1% triethanolamine, 0.5%-1% sodium lauryl sulfate, 8%-12% glycerin, and the balance being water; wherein the drug is composed of Artemisia argyi oil, egg yolk oil, and comfrey oil in a weight ratio of 1-5:1-4:1-5. The egg yolk oil is prepared by the following method: boil eggs, remove egg whites, grind egg yolks into powder, place them in a stainless steel pot, heat over low heat until at least a small amount of oil is produced, stir-fry over high heat until the egg yolks turn brown, filter, remove the residue, and the oil is obtained. The comfrey oil is prepared by the following method: weigh 6 times the weight of comfrey in rapeseed oil, heat it to boiling, add the comfrey, extract for 5 minutes, filter, and the oil is obtained.

2. The Vernonia amygdalina cream according to claim 1, wherein, The formulation of the Artemisia argyi cream consists of the following components by weight percentage: 9% medicine, 5% white petrolatum, 5% stearic acid, 5% glyceryl monostearate, 10% liquid paraffin, 0.2% ethylparaben, 4% Tween-80, 0.5% triethanolamine, 0.5% sodium lauryl sulfate, 10% glycerin, and the balance being water; wherein the medicine is composed of Artemisia argyi oil, egg yolk oil, and comfrey oil in a weight ratio of 1:1:

1.

3. The method for preparing Artemisia argyi cream as described in claim 1, characterized in that, The specific steps are as follows: accurately weigh each component according to the formula; add senna oil, egg yolk oil, comfrey oil, white petrolatum, stearic acid, glyceryl monostearate, liquid paraffin, and ethylparaben to the oil phase beaker and heat in an 80°C water bath until completely dissolved; add glycerin, Tween-80, triethanolamine, sodium lauryl sulfate, and water to the aqueous phase beaker and heat in an 80°C water bath until completely dissolved; slowly add the aqueous phase to the oil phase and emulsify at 300-700 rpm and 60°C-80°C for 5-15 minutes, stirring continuously and cooling to room temperature to obtain senna cream.

4. The method for preparing Artemisia argyi cream according to claim 3, characterized in that, The stirring speed for emulsification is 500-700 rpm.

5. The method for preparing Artemisia argyi cream according to claim 4, characterized in that, The stirring speed for emulsification is 500 rpm.

6. The method for preparing Artemisia argyi cream according to claim 3, characterized in that, The emulsification temperature is 70℃-80℃.

7. The method for preparing Artemisia argyi cream according to claim 6, characterized in that, The emulsification temperature is 80°C.

8. The method for preparing Artemisia argyi cream according to claim 3, characterized in that, The emulsification time is 5-10 minutes.

9. The method for preparing Artemisia argyi cream according to claim 8, characterized in that, The emulsification time is 5 minutes.

10. The use of the Artemisia argyi cream as described in claim 1 in the preparation of a drug for promoting the repair of Grade II burn wounds.

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