Licochalcone A gel paste with anti-inflammatory and analgesic functions and preparation method thereof

Abstract

The invention discloses a licochalcone A gel paste with anti-inflammatory and analgesic functions and a preparation method thereof, and relates to the technical field of medicines, the licochalcone A gel paste comprises the following components by mass percentage: 0.5%-1.5% of carbomer 940, 5%-10% of glycerin, 1%-2% of a pH regulator, 87%-92% of licochalcone A, and the balance of water. The licochalcone A gel paste is safe and effective, has good antibacterial and anti-inflammatory performance, belongs to a natural medicine, can effectively promote healing of second-degree burn wounds, can further deeply analyze a molecular mechanism of wound repair, and has important clinical significance and academic value for improving the current situation of burn treatment.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions and its preparation method. Background Technology

[0002] Burns are a common traumatic disease. The treatment of second-degree burns is complex and lengthy, involving multiple stages such as inflammation, proliferation, and remodeling, and requires delicate cellular and molecular interactions.

[0003] Currently, burn treatment methods mainly include topical medications, antibiotic therapy, surgical repair, and wound care products. However, these traditional methods have many shortcomings, such as: 1. Significant issues with drug toxicity: Many topical medications have a certain degree of toxicity, and long-term use may cause serious side effects to the patient's body. In addition, some medications pose significant safety risks.

[0004] 2. Limited anti-inflammatory and healing effects: Existing drugs have extremely limited effects in controlling inflammatory responses and promoting wound healing, which cannot meet the urgent clinical needs, resulting in slow treatment progress and seriously affecting patients' quality of life.

[0005] 3. Poor antibacterial properties: Some topical medications have weak inhibitory effects on common pathogens, which can easily lead to wound infection and increase treatment risks.

[0006] 4. Lack of comprehensive treatment options: Existing treatment methods fail to simultaneously address multiple functions such as antibacterial, anti-inflammatory, and collagen deposition promotion, and the molecular mechanisms of wound healing are not well understood, making it difficult to specifically regulate the healing process.

[0007] Given the aforementioned limitations, there is an urgent need to develop a safe, effective, and effective natural drug with good antibacterial and anti-inflammatory properties. Summary of the Invention

[0008] Given the problems of significant toxicity, limited anti-inflammatory and antibacterial effects, and poor healing efficacy of drugs used in current burn treatment methods, the purpose of this invention is to provide a glycyrrhizin chalcone A gel ointment with anti-inflammatory and analgesic functions, its preparation method, and its application. This glycyrrhizin chalcone A gel ointment is safe and effective, and has good antibacterial and anti-inflammatory properties. As a natural drug, it can not only effectively promote the healing of second-degree burn wounds, but also deeply elucidate the molecular mechanism of wound repair, which has important clinical significance and academic value for improving the current status of burn treatment.

[0009] This invention is achieved through the following technical solution: In one aspect, this application provides a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions, comprising, by weight percentage, 0.5% to 1.5% carbomer 940, 5% to 10% glycerin, 1% to 2% pH adjuster, 87% to 92% glycyrrhizin chalcone A, and the balance being water.

[0010] Furthermore, the pH value of the glycyrrhizin chalcone A gel is 6.5-7.5. Preferably, the pH value of the glycyrrhizin chalcone A gel is 7.

[0011] Furthermore, the pH adjuster includes triethanolamine.

[0012] Secondly, this application provides a method for preparing glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions, comprising the following steps: Dissolve Carbomer 940 in water; Add glycerin and a pH adjuster to adjust viscosity and pH value; After dissolving in glycyrrhizin A, dilute to volume with water.

[0013] Furthermore, the viscosity is adjusted to 130 mPa·s to 190 mPa·s. Preferably, the viscosity is adjusted to 140 mPa·s.

[0014] Thirdly, this application provides a medicament for treating burns, including the above-mentioned glycyrrhizin chalcone A gel ointment or glycyrrhizin chalcone A gel ointment prepared by the above-mentioned preparation method.

[0015] Fourthly, this application provides a gel plaster, including the above-mentioned glycyrrhizin A gel plaster or glycyrrhizin A gel plaster prepared by the above-mentioned preparation method.

[0016] Fifthly, this application provides a traditional Chinese medicine patch, including the aforementioned gel plaster.

[0017] Compared with the prior art, the present invention has the following advantages and beneficial effects: (1) The glycyrrhizin chalcone A gel ointment of the present invention is safe and effective, and has good antibacterial and anti-inflammatory properties. It is a natural drug that can not only effectively promote the healing of second-degree burn wounds, but also deeply analyze the molecular mechanism of wound repair. It has important clinical significance and academic value for improving the current status of burn treatment.

[0018] (2) The glycyrrhizin A gel paste of the present invention does not exhibit stratification, flocculation or liquefaction in high temperature and low temperature environments. It has a smooth, uniform and delicate surface, good spreadability, and can be spread quickly.

[0019] (3) The glycyrrhizin chalcone A gel ointment of the present invention has good moisturizing and anti-inflammatory effects, and can effectively shorten the skin healing time, accelerate the healing speed, promote collagen repair of burn tissue, reduce inflammatory infiltration after burns, and protect tissue from excessive infiltration of inflammatory factors that lead to further development of tissue wounds.

[0020] (4) The licorice chalcone A gel ointment of the present invention has strong antibacterial activity against Staphylococcus epidermidis and Escherichia coli, and can effectively inhibit their growth.

[0021] (5) The glycyrrhizin chalcone A gel ointment of the present invention has no damaging effect on the tissue structure of important organs during continuous use, indicating that it has good safety.

[0022] (6) The glycyrrhizin chalcone A gel ointment of the present invention has a significant protective effect against cellular oxidative stress, can protect fibroblasts from apoptosis, can protect the cell migration ability, and can play an effective protective role in the burn healing process. Attached Figure Description

[0023] To more clearly illustrate the technical solutions of the exemplary embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly described below. It should be understood that the following drawings only show some embodiments of the present invention and should not be considered as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort. In the drawings: Figure 1 This is a graph showing the high-temperature stability test results of the glycyrrhizin chalcone A gel prepared in the embodiments of the present invention; Figure 2 This is a graph showing the low-temperature stability test results of the glycyrrhizin chalcone A gel prepared in the embodiments of the present invention; Figure 3 The image shows the results of the moisturizing test of the glycyrrhizin chalcone A gel prepared in the embodiments of the present invention; Figure 4 The image shows the results of the antibacterial experiment using the paper disc method on the glycyrrhizin chalcone A gel prepared in this embodiment of the invention, where A represents Staphylococcus epidermidis; B represents Escherichia coli; C represents Staphylococcus aureus; and D represents Pseudomonas aeruginosa. Figure 5 This is a diagram showing the effect of a mouse ear swelling test in an embodiment of the present invention; Figure 6 HE staining images of tissue sections from important organs of mice in this embodiment of the invention; Figure 7 This is a diagram illustrating the burn healing process in mice, as described in an embodiment of the present invention. Figure 8This is a statistical result diagram of scalded wounds in mice in an embodiment of the present invention; Figure 9 This is a graph showing the relative collagen content of each treatment group in Sirius red staining in an embodiment of the present invention; Figure 10 This is a diagram showing the effect of LCA on collagen in 6-day and 9-day scalded skin samples from mice in this embodiment of the invention. Figure 11 This is a statistical chart showing the relative collagen content of each treatment group in the masson staining process in this embodiment of the invention. Figure 12 This is a graph showing the effect of LCA on inflammatory factors IL-6, TNF-α, and IL-1β in mouse skin samples 6 days after burns, as described in this embodiment of the invention. Figure 13 This is a statistical graph showing the effect of LCA on the inflammatory factors IL-6, TNF-α, and IL-1β in mouse skin samples 6 days after burns, as described in this embodiment of the invention. Figure 14 This is a graph showing the effect of LCA on Ly6G and F4 / 80 in mouse skin samples 6 days after burns, as described in this embodiment of the invention. Figure 15 This is a statistical graph showing the effect of LCA on Ly6G and F4 / 80 in mouse skin samples 6 days after burns, as described in this embodiment of the invention. Figure 16 This is a graph showing the effect of LCA on Ki67 and VEGF-C in mouse skin samples 6 days after burns, as described in this embodiment of the invention. Figure 17 This is a statistical graph showing the effect of LCA on Ki67 and VEGF-C in mouse skin samples 6 days after burns, as described in this embodiment of the invention. Figure 18 The graphs show the cytotoxicity of different concentrations of glycyrrhizin A on cells in this embodiment of the invention; where A is a bar chart of cytotoxicity in L929 cells; and B is a bar chart of cytotoxicity in A9 cells. Figure 19 The graphs show the cytotoxicity of different concentrations of hydrogen peroxide to cells in this embodiment of the invention; where A is a bar chart of cytotoxicity of L929 cells and B is a bar chart of cytotoxicity of A9 cells. Figure 20 This is a graph showing the protective effect of LCA against cellular oxidative stress in an embodiment of the present invention; where A represents L929 cells and B represents A9 cells; Figure 21 This is a graph showing the effect of LCA on ROS in L929 and A9 cells in an embodiment of the present invention; where A represents L929 cells and B represents A9 cells. Figure 22This is a statistical graph showing the relative fluorescence intensity of ROS in L929 and A9 cells in an embodiment of the present invention; where A represents L929 cells and B represents A9 cells. Figure 23 This is a diagram illustrating the effect of LCA on apoptosis in L929 cells in an embodiment of the present invention. Figure 24 This is a diagram illustrating the effect of LCA on apoptosis in A9 cells in an embodiment of the present invention. Figure 25 This is a diagram illustrating the effect of LCA on cell migration ability in L929 cells in an embodiment of the present invention. Figure 26 This is a diagram illustrating the effect of LCA on cell migration ability in A9 cells in an embodiment of the present invention. Figure 27 This is a statistical analysis diagram of Transwell in L929 and A9 cells in an embodiment of the present invention; where A represents L929 cells and B represents A9 cells; Figure 28 This is a graph showing the effect of LCA on the oxidative index levels of L929 cells in an embodiment of the present invention; where A represents the cell GSH-PX level, B represents the cell CAT level, and C represents the cell LDH level. Figure 29 This is a diagram showing the effect of LCA on the oxidative index levels of A9 cells in an embodiment of the present invention; where A represents the cell LDH level, B represents the cell GSH-PX level, and C represents the cell CAT level. Figure 30 This is a diagram showing the effect of LCA on the levels of TNF-α, IL-1β and IL-6 in L929 cells in an embodiment of the present invention, where A represents the level of TNF-α in cells, B represents the level of IL-1β in cells, and C represents the level of IL-6 in cells; Figure 31 This diagram illustrates the effect of LCA on the levels of TNF-α, IL-1β, and IL-6 in A9 cells in an embodiment of the present invention. In this diagram, A represents the level of TNF-α in the cells, B represents the level of IL-1β in the cells, and C represents the level of IL-6 in the cells.

[0024] Figure 32 These are electron micrographs of L929 cells under different treatment groups in this embodiment of the invention; Figure 33 The graph shows the test results of the high temperature stability, low temperature stability, and room temperature moisturizing properties of the gel paste in Comparative Example 1 of the present invention. Figure 34 The graph shows the test results of the high temperature stability, low temperature stability, and room temperature moisturizing properties of the gel paste in Comparative Example 2 of the present invention. Figure 35The graph shows the test results of the high temperature stability, low temperature stability, and room temperature moisturizing properties of the gel paste in Comparative Example 3 of this invention. Detailed Implementation

[0025] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the embodiments. The illustrative embodiments and descriptions of this invention are only used to explain this invention and are not intended to limit this invention.

[0026] In the following description, numerous specific details are set forth in order to provide a thorough understanding of the invention. However, it will be apparent to those skilled in the art that these specific details are not necessary to practice the invention. In other embodiments, well-known materials or methods have not been specifically described in order to avoid obscuring the invention.

[0027] Throughout this specification, references to “an embodiment,” “an example,” or “an example” mean that a particular feature, structure, or characteristic described in connection with that embodiment or example is included in at least one embodiment of the invention. Therefore, the phrases “an embodiment,” “an example,” “an example,” or “an example” appearing in various places throughout the specification do not necessarily refer to the same embodiment or example. Furthermore, specific features, structures, or characteristics can be combined in one or more embodiments or examples in any suitable combination and / or sub-combination. The term “and / or” as used herein includes any and all combinations of one or more of the associated listed items.

[0028] The "range" disclosed in this application is defined by a lower limit and an upper limit. A given range is defined by selecting a lower limit and an upper limit, which define the boundaries of a particular range. Ranges defined in this way can include or exclude endpoints and can be arbitrarily combined; that is, any lower limit can be combined with any upper limit to form a range. For example, if ranges of 60–120 and 80–110 are listed for a specific parameter, it is understood that ranges of 60–110 and 80–120 are also expected. Furthermore, if minimum range values ​​of 1 and 2 are listed, and if maximum range values ​​of 3, 4, and 5 are listed, then the following ranges are all expected: 1–3, 1–4, 1–5, 2–3, 2–4, and 2–5. In this application, unless otherwise stated, the numerical range "a–b" represents a shortened representation of any combination of real numbers between a and b, where a and b are real numbers. For example, the numerical range "0~5" indicates that all real numbers between "0~5" have been listed in this article; "0~5" is simply a shortened representation of these numerical combinations. Furthermore, when a parameter is stated as an integer ≥2, it is equivalent to disclosing that the parameter is, for example, an integer such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, etc.

[0029] Unless otherwise specified, all steps in this application may be performed sequentially or randomly, preferably sequentially. For example, the method includes steps (a) and (b), indicating that the method may include steps (a) and (b) performed sequentially, or it may include steps (b) and (a) performed sequentially. For example, the method may also include step (c), indicating that step (c) may be added to the method in any order. For example, the method may include steps (a), (b), and (c), or it may include steps (a), (c), and (b), or it may include steps (c), (a), and (b), etc.

[0030] Example 1 This embodiment provides a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions and its preparation method. The formula of the glycyrrhizin chalcone A gel is: 1.245g of carbomer 940, 8.713g of glycerin, 1.5g of triethanolamine, 200umol of glycyrrhizin chalcone A, and the balance is water.

[0031] The preparation method is as follows: S1. Dissolve Carbomer 940 in water; S2. Add glycerol and triethanolamine to adjust the viscosity to 140 mPa·s and the pH to 7; S3. After dissolving in glycyrrhizin A, dilute to 100ml with water.

[0032] Example 2 This embodiment provides a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions and its preparation method. Unlike Example 1, the formulation of the glycyrrhizin chalcone A gel in this embodiment is: 0.5g of carbomer 940, 5g of glycerin, 1g of triethanolamine, 100umol of glycyrrhizin chalcone A, and the balance is water.

[0033] The preparation method is the same as in Example 1.

[0034] Example 3 This embodiment provides a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions and its preparation method. Unlike embodiment 1, the formulation of the glycyrrhizin chalcone A gel in this embodiment is: 1.5g of carbomer 940, 10g of glycerin, 2g of triethanolamine, 300umol of glycyrrhizin chalcone A, and the balance is water.

[0035] The preparation method is the same as in Example 1.

[0036] Example 4 This embodiment provides a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions and its preparation method. Unlike Example 1, the pH value of the glycyrrhizin chalcone A gel in this embodiment is 6.5. Its formulation is the same as in Example 1.

[0037] The preparation method is as follows: S1. Dissolve Carbomer 940 in water; S2, add glycerol and triethanolamine to adjust the viscosity to 140 mPa·s and the pH to 6.5; S3. After dissolving in glycyrrhizin A, dilute to 100ml with water.

[0038] Example 5 This embodiment provides a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions and its preparation method. Unlike Example 1, the pH value of the glycyrrhizin chalcone A gel in this embodiment is 7.5. Its formulation is the same as in Example 1.

[0039] The preparation method is as follows: S1. Dissolve Carbomer 940 in water; S2, add glycerol and triethanolamine to adjust the viscosity to 140 mPa·s and the pH to 6.5; S3. After dissolving in glycyrrhizin A, dilute to 100ml with water.

[0040] Example 6 This embodiment provides a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions and its preparation method. The difference from Example 1 is that the viscosity in this embodiment is adjusted to 130 mPa·s. Its formulation is the same as in Example 1.

[0041] The preparation method is as follows: S1. Dissolve Carbomer 940 in water; S2, add glycerol and triethanolamine to adjust the viscosity to 130 mPa·s and the pH to 6.5; S3. After dissolving in glycyrrhizin A, dilute to 100ml with water.

[0042] Example 7 This embodiment provides a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions and its preparation method. The difference from Example 1 is that the viscosity in this embodiment is adjusted to 190 mPa·s. Its formulation is the same as in Example 1.

[0043] The preparation method is as follows: S1. Dissolve Carbomer 940 in water; S2, add glycerol and triethanolamine to adjust the viscosity to 190 mPa·s and the pH to 6.5; S3. After dissolving in glycyrrhizin A, dilute to 100ml with water.

[0044] Comparative Example 1 This comparative example provides a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions and its preparation method. Unlike Example 1, the formulation of this comparative example does not contain carbomer 940. The formulation of its glycyrrhizin chalcone A gel is: 8.713g of glycerin, 1.5g of triethanolamine, 200umol of glycyrrhizin chalcone A, and the balance is water.

[0045] The preparation method is as follows: S1. Dissolve glycyrrhizin A in water; S2. Add glycerol and triethanolamine to adjust the viscosity to 140 mPa.s and the pH to 7, then dilute to 100 ml with water.

[0046] Comparative Example 2 This embodiment provides a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions and its preparation method. Unlike Example 1, the formulation of this comparative example does not contain glycerin. The formulation of the glycyrrhizin chalcone A gel is: 1.245g of carbomer 940, 1.5g of triethanolamine, 200umol of glycyrrhizin chalcone A, and the balance is water.

[0047] The preparation method is as follows: S1. Dissolve Carbomer 940 in water; S2. Add triethanolamine to adjust the viscosity to 140 mPa·s and the pH to 7; S3. After dissolving in glycyrrhizin A, dilute to 100ml with water.

[0048] Comparative Example 3 This comparative example provides a glycyrrhizin chalcone A gel with anti-inflammatory and analgesic functions and its preparation method. Unlike Example 1, the formulation of this comparative example does not contain carbomer 940 and glycerin. The formulation of its glycyrrhizin chalcone A gel is: 1.5g of triethanolamine, 200umol of glycyrrhizin chalcone A, and the balance is water.

[0049] The preparation method is as follows: S1. Dissolve glycyrrhizin A in water; S2. Add triethanolamine to adjust the viscosity to 140 mPa.s and the pH to 7, then dilute with water to 100 ml.

[0050] The properties of the glycyrrhizin chalcone A gel prepared using the methods of Examples 1-7 and Comparative Examples 1-3 were tested as follows.

[0051] Performance Test 1: The hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 were subjected to high-temperature stability tests.

[0052] Approximately 5.0 g of LCA hydrogel was weighed and placed in a transparent container, sealed, and placed in a 60°C incubator for 12 hours. After cooling to room temperature, the state of the gel was observed. The spreadability of the gel was tested from its surface and sides. The test results for Example 1 are as follows: Figure 1 As shown, the detection results of other embodiments are shown in the figure. Figure 1 The results were essentially the same, with no stratification, flocculation, or liquefaction observed. The surfaces were smooth, uniform, and fine, exhibiting good spreadability and rapid dispersibility. This indicates that the LCA hydrogel is relatively stable in high-temperature environments within 12 hours.

[0053] from Figure 33 As can be seen, Comparative Example 1, lacking Carbomer 940 as a gel matrix, is in a liquid state and exhibits obvious stratification after 12 hours at 60℃. The upper layer is a clear liquid, while the lower layer contains precipitates, making it unable to form a stable gel system. It has poor spreadability and excessive fluidity.

[0054] from Figure 34 It can be seen that the gel in Comparative Example 2, due to the lack of glycerin and excessive viscosity, exhibited cracking and delamination after 12 hours at a high temperature of 60℃, which seriously affected the spreadability of the gel.

[0055] from Figure 35 As can be seen, Comparative Example 3, lacking both carbomer 940 and glycerin, consisted only of an aqueous solution containing glycyrrhizin chalcone A and triethanolamine. After 12 hours at 60°C, the system remained in a liquid state, exhibiting extremely poor stability and failing to maintain its shape, thus lacking the basic properties required for a gel dressing. Drug precipitation significantly impacted the sustained-release effect of the gel.

[0056] Performance Test 2: The hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 were subjected to low-temperature stability tests.

[0057] Approximately 5.0 g of LCA hydrogel was weighed and placed in a transparent container, sealed, and stored in a -20 ℃ refrigerator for 12 hours. After cooling to room temperature, the state of the gel was observed. The test results of Example 1 are as follows: Figure 2 As shown, the detection results of other embodiments are shown in the figure. Figure 2 The results were essentially the same. Observation and spreadability tests of the surface and sides of the gel showed no delamination, flocculation, or liquefaction. The surface was smooth, uniform, and fine, with good spreadability and rapid spreading. This indicates that the LCA hydrogel is relatively stable in a low-temperature environment within 12 hours.

[0058] from Figure 33It can be seen that after 12 hours at a low temperature of -20℃, Comparative Example 1 showed stratification and partial crystallization due to the lack of a gel matrix. After returning to room temperature, the liquid was still turbid and the system was not uniform.

[0059] from Figure 34 It can be seen that, after 12 hours at a low temperature of -20℃, the yellow color of Comparative Example 2 faded due to the lack of antifreeze effect of glycerin, and flocculation and stratification occurred. It also had poor spreadability and could not be spread quickly.

[0060] from Figure 35 It can be seen that after 12 hours at -20℃, the liquid in Comparative Example 3 froze, and after returning to room temperature, obvious phase separation occurred, with glycyrrhizin A precipitating and crystallizing, and the system completely losing its homogeneity.

[0061] Performance Test 3: The hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 were subjected to moisturizing tests.

[0062] Weigh 5g of LCA hydrogel and place it in a 60mm petri dish. Incubate at 37°C for 12 and 24 hours under ventilated conditions. The test results of Example 1 are as follows: Figure 3 As shown, the detection results of other embodiments are shown in the figure. Figure 3 Basically the same, from Figure 3 As can be seen above, the measured weight loss was 3.99% and 4.19%, indicating that glycyrrhizin chalcone A gel has good moisturizing properties.

[0063] from Figure 33 It can be seen that the gel of Comparative Example 1, after being placed at a constant temperature of 37℃ for 12 hours, partially lost moisture after 24 hours, with a weight loss of 6.59% and 11.56%, indicating poor moisturizing properties.

[0064] from Figure 34 It can be seen that although Comparative Example 2 can form a gel, it lacks glycerin as a moisturizer. After being placed in a constant temperature incubator at 37℃ for 12h and 24h, the weight loss rates were 7.93% and 15.41%, respectively, and the moisturizing performance was significantly worse than that of the example containing glycerin.

[0065] from Figure 35 It can be seen that Comparative Example 3, after being placed at a constant temperature of 37℃ for 12 hours, showed a significant weight loss after 24 hours. As a liquid system, the weight loss rates were 6.31% and 12.54%, respectively, which did not have the moisturizing properties required for gel dressings.

[0066] Performance Test 4: The hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 were subjected to antibacterial tests.

[0067] The hydrogel was spread onto a culture plate, different bacterial strains were inoculated, and the results were observed after incubation. The detection results of Example 1 are as follows: Figure 4 As shown, the detection results of other embodiments are shown in the figure. Figure 4 Basically the same, such as Figure 4 The minimum bactericidal concentration (MCC) of glycyrrhizin A against Staphylococcus epidermidis and Escherichia coli was 250 μM, while the MCC against Staphylococcus aureus and Pseudomonas aeruginosa was 500 μM. This result indicates that glycyrrhizin A possesses strong antibacterial activity against Staphylococcus epidermidis and Escherichia coli, effectively inhibiting their growth. In contrast, Staphylococcus aureus and Pseudomonas aeruginosa showed lower sensitivity to glycyrrhizin A, demonstrating the difference in its antibacterial efficacy.

[0068] Performance Test 5: Mouse Ear Swelling Test to Verify the Anti-inflammatory Properties of the Hydrogel Dressings Prepared in Examples 1-7 and Comparative Examples 1-3 C57BL6J mice were selected as experimental animals. Mice in the glycyrrhizin-chalcone A group had 100 μM glycyrrhizin-chalcone A gel applied to their ears daily for two days prior to treatment, while the control group received physiological saline. In the experiment, 50 μL of xylene was evenly applied to both sides of the left auricle of the mice, while the right ear was left untreated as a control, thus establishing an acute ear swelling model. Forty-five minutes after treatment, the mice were euthanized by cervical dislocation, and both ears were immediately removed. Ear flaps were taken from the same location using a 6 mm punch and weighed to analyze the degree of swelling. The swelling degree was calculated as the weight of the left ear flap minus the weight of the right ear flap, and the swelling rate was calculated as (left ear weight - right ear weight) / right ear weight. The experimental results showed that the degree and rate of ear swelling in the model group were higher than those in the treatment group. The results of Example 1 are as follows: Figure 5 As shown, the detection results of other embodiments are shown in the figure. Figure 5 Basically the same, in Figure 5 In the study, the ear swelling rate in the treated group was significantly lower than that in the model group (P<0.05). These results indicate that glycyrrhizin A can effectively inhibit ear swelling in mice, demonstrating its anti-inflammatory effect.

[0069] Performance Test 6: Safety Assessment of the Hydrogel Dressings Prepared in Examples 1-7 and Comparative Examples 1-3 Safety assessment of LCA hydrogel was conducted in C57 mice. Glycyrrhizin chalcone A gel (n=10) was applied topically to the backs of mice once daily for 15 consecutive days. Behavior, diet, and health status of the mice were observed. After administration, blood samples were collected for liver function tests. Samples were stored at 4 °C and tested within 30 minutes if possible; otherwise, they were centrifuged and stored at -20 °C in a light-protected, sealed container. Mice were then euthanized by cervical dislocation, and samples from vital organs such as the heart, liver, spleen, lungs, kidneys, stomach, and intestines were extracted, embedded, and cut into 8 μm sections for HE staining and pathological observation. The results of Example 1 are as follows: Figure 6 As shown, the detection results of other embodiments are shown in the figure. Figure 6Basically the same, Figure 6 The results showed that glycyrrhizin A did not damage the tissue structure of any important organs in mice during continuous administration, indicating that it has good safety.

[0070] Performance Test 7: Effect of the hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 on the skin healing time of burned mice. This study selected 45 C57 mice aged 6-8 weeks and randomly divided them into a blank control group, a glycyrrhizin-chalcone A group, and a model group, with 15 mice in each group. After hair removal using a 6% sodium sulfide dilution, a self-made burn instrument was used to apply a 100°C heat source to the hairless area on the back of the mice for 5 seconds to establish a second-degree burn model. Before wound creation, the mice were anesthetized with 5% chloral hydrate. Subsequently, different medications were applied to the burn wounds daily, and photographs were taken on days 1, 3, 6, 9, and 13 to analyze the burn area and healing rate. The healing rate was calculated as (original area - unhealed area) / original area. Samples were taken on days 3, 5, and 7. Mice were euthanized by cervical dislocation, strictly adhering to animal ethics protection laws. After disinfection, skin grafts were separated from the wounds and histologically processed. The results of Example 1 are as follows. Figure 7 and Figure 8 As shown, the detection results of other embodiments are shown in the figure. Figure 7 and Figure 8 Basically the same, Figure 7 and Figure 8 The results showed that the burned mice were in good general condition, with roughly the same wound area, localized dryness, and circular white borders around the wound edges. On day 6, the wound area in the LCA group mice was significantly reduced, with the appearance of light red new epithelial tissue, an improvement compared to day 3. On day 9, all wounds continued to shrink, scabs partially fell off, exposing pink new skin, and most wounds had become epithelialized. The LCA group showed a significant healing advantage compared to other groups. This indicates that glycyrrhizin chalcone A gel ointment can effectively shorten the skin healing time in mice.

[0071] Performance Test 8: The hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 promoted collagen repair in burn tissues. Sirius red staining is primarily used to observe and evaluate collagen fibers in tissues. By observing the color, arrangement, and distribution of collagen fibers, the degree of fibrosis and collagen metabolism in the tissue can be assessed. The detection results of Example 1 are as follows... Figure 9 As shown, the detection results of other embodiments are shown in the figure. Figure 9 Basically the same, from Figure 9As can be seen, at 6 days, fewer yellow / orange-red collagen fibers were observed in the model group, indicating a lower content of type I collagen fibers. The overall staining was darker. In the treatment group, the larger red area indicated a higher collagen content. In the control group, the collagen fibers were stained relatively evenly and the color was lighter, suggesting that the collagen content and arrangement may be close to normal.

[0072] At day 9, the yellow / orange-red area appeared to increase in the model group compared to day 6, possibly indicating increased deposition of type I collagen fibers. In the treatment group, staining was deeper, but the collagen-stained area was larger and collagen deposition more pronounced compared to the model group. In the control group, collagen fiber staining was relatively uniform and the color was lighter, suggesting that collagen content and arrangement may be close to normal. Statistical analysis demonstrated that LCA gel can promote collagen proliferation and accelerate healing.

[0073] Performance Test 9: The hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 promoted collagen repair in burn tissue. Masson's trichrome stain is a commonly used histological staining method to distinguish different components in tissues. It primarily uses a combination of three dyes to make collagen fibers appear blue, muscle, erythrocytes, and keratin appear red, and cell nuclei appear dark blue or black. Masson's stain is frequently used to observe and assess the degree of fibrosis in tissues, as well as to differentiate components such as muscle and collagen. The detection results in Example 1 are as follows... Figure 10 As shown, the detection results of other embodiments are shown in the figure. Figure 10 Basically the same, from Figure 10 As can be seen, at 6 days, collagen fibers in the model group were dispersed and broken, while collagen in the treatment group was more dense, and there was a significant difference between the model group and the control group. At 9 days, collagen fragmentation was still observed in the dermis of the model group, while collagen in the treatment group was more tightly arranged and had a larger collagen distribution area. Figure 8 The bar chart also shows that the collagen content in the treatment group was higher than that in the model group, and the difference was significant, confirming that glycyrrhizin A can promote collagen synthesis.

[0074] Performance Test 10: The hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 inhibited excessive inflammatory response. To investigate the effects of LCA on inflammatory factors and their proliferation during skin healing in mice, we performed immunohistochemical staining on skin tissue sections from each treatment group on day 6. The results of Example 1 are as follows: Figures 11-16 As shown, the detection results of other embodiments are shown in the figure. Figures 11-16 The distribution of inflammatory factors (IL-6, TNF-α, IL-1β) is basically the same as that of other factors. Figure 11-12As shown, the distribution of inflammatory factors in the LCA group was significantly lower than that in the model group, which can also be seen from the statistical bar chart. The distribution of neutrophils and macrophages was also observed. Figure 13-14 It was observed that the LCA group had less infiltration of neutrophils and macrophages, while the model group had extensive infiltration of neutrophils and macrophages, significantly more than the control group and the LCA group. Mouse skin proliferation was determined by... Figure 15-16 As can be seen, the number of Ki67 and VEGF-positive cells was significantly increased in the LCA group, higher than in the model group and the control group. This indicates that glycyrrhizin A can reduce inflammatory infiltration after burns and protect tissues from excessive infiltration of inflammatory factors that could lead to further wound development. Simultaneously, glycyrrhizin A can promote tissue proliferation and accelerate wound healing.

[0075] Performance Test 11: Cytotoxicity Test of Hydrogel Dressings Prepared in Examples 1-7 and Comparative Examples 1-3 Cells were stimulated for 24 h with glycyrrhizin A at concentrations of 0, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM, 35 μM, and 40 μM, respectively, and cell viability was then detected using a CCK-8 assay kit. The results of Example 1 are shown below. Figure 17 As shown, the detection results of other embodiments are shown in the figure. Figure 17 Basically the same, the results are as follows Figure 17 As shown in Figure A, the cell viability of L929 cells after 24 hours of LCA stimulation is approximately 66% at a concentration of 20 μM, but decreases sharply at 30 μM. Therefore, 20 μM should be used as the experimental concentration. Similarly, the cell viability of A9 cells drops below 50% in a concentration gradient of 30-35 μM, so 30 μM should be used as the treatment concentration for subsequent experiments.

[0076] Performance Test 12: Construction of Cell Oxidative Stress Models for Hydrogel Dressings Prepared in Examples 1-7 and Comparative Examples 1-3 Cells were stimulated for 12 h with hydrogen peroxide solutions of 0, 400 μM, 600 μM, 800 μM, 1000 μM, 1200 μM, and 1400 μM, respectively. The detection results in Example 1 are as follows: Figure 18 As shown, the detection results of other embodiments are shown in the figure. Figure 18 Basically the same, the results are as follows Figure 18 Figure A shows the cell viability of L929 cells after 12 h of hydrogen peroxide stimulation. At a concentration of 1000 μM, the viability was approximately 60%, but it decreased sharply at 1200 μM. Therefore, 950 μM was chosen as the experimental concentration. Similarly, the viability of A9 cells dropped below 50% in a concentration gradient of 1200-1400 μM; therefore, 1200 μM should be selected as the treatment concentration for subsequent experiments.

[0077] Performance Test 13: Protective Effect of Hydrogel Dressings Prepared in Examples 1-7 and Comparative Examples 1-3 on Cellular Oxidative Stress L929 and A9 cells were preprotected with 20 μM and 30 μM glycyrrhizin A, respectively, for 24 h. Then, hydrogen peroxide solutions of the corresponding concentrations described in section 3.2.7 were added. After 12 h, cell viability of both groups was detected using a CCK-8 assay kit. The results of Example 1 are shown below. Figure 19 As shown, the detection results of other embodiments are shown in the figure. Figure 19 Basically the same, from Figure 19 As can be seen above, after pre-protection, the cell viability of both drug-treated cell lines was significantly higher than that of the model group (P<0.05). The results indicate that glycyrrhizin A has a significant protective effect against oxidative stress induced by hydrogen peroxide.

[0078] Performance Test 14: Effects of the hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 on cellular ROS ROS are a class of highly reactive oxygen-containing molecules that play a role in cell signaling and homeostasis. However, excessive ROS can lead to oxidative stress, damaging cellular structures, including DNA, proteins, and lipids, and consequently causing various diseases. The detection results in Example 1 are as follows... Figures 21-22 As shown, the detection results of other embodiments are shown in the figure. Figures 21-22 Basically the same, the effect of glycyrrhizin A on ROS is as follows: Figure 21 As shown in the figure, green fluorescence represents ROS. The fluorescence area and intensity of the LCA-treated group were significantly lower than those of the model group, indicating that LCA effectively reduced the production of intracellular ROS induced by hydrogen peroxide, protecting cells from reactive oxygen species damage. The bar chart also confirms this conclusion. Figure 22 As can be seen, there was a significant difference in the relative fluorescence intensity between the LCA group and the model group. LCA has a significant protective effect against hydrogen peroxide-induced oxidative stress.

[0079] Performance Test 15: The hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 inhibited cell apoptosis. Apoptosis is one of the most direct indicators reflecting the protective effect of drugs. The above experiments have demonstrated that LCA has a protective effect on cells and can protect cells from damage caused by hydrogen peroxide. After pre-protection and hydrogen peroxide stimulation, cells were stained using a live / dead cell double staining kit and examined under a fluorescence microscope. The results are as follows. The detection results of Example 1 are as follows: Figures 23-24 As shown, the detection results of other embodiments are shown in the figure. Figures 23-24 Basically the same, according to Figure 23As can be seen, the model group exhibited strong red fluorescence and had a predominance of red cells, indicating that most cells in the model group underwent apoptosis under hydrogen peroxide stimulation. In contrast, the LCA group showed fewer red cells, and the cells remained morphologically intact and in good condition, suggesting that LCA has a protective effect. Figure 24 Similarly, the number of apoptotic cells in the LCA group was significantly lower than that in the model group. Cells are damaged and undergo apoptosis when stimulated by hydrogen peroxide. Glycyrrhizin chalcone A can protect fibroblasts from apoptosis, playing an effective protective role in the burn healing process and promoting faster wound healing.

[0080] Performance Test 16: The ability of hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 to inhibit cell migration The migration ability of fibroblasts significantly affects the healing speed of burn wounds. This experiment investigated the migration ability of cells after LCA preprotection. The results of Example 1 are as follows: Figures 25-27 As shown, the detection results of other embodiments are shown in the figure. Figures 25-27 Basically the same, such as Figure 25 , 26 As shown, the control group had the highest number of cells that passed through the Transwell chamber, followed by the LCA group. Compared to the model group, the LCA group had significantly more cells that passed through and better cell morphology, while the model group, due to hydrogen peroxide stimulation and the lack of drug protection, had a very limited number of cells that passed through. The chamber was placed in ethanol to dissolve the stained crystal violet, and its absorbance was measured at 595 nm; this value represents the number of cells that passed through the chamber. Figure 27 It is evident that the LCA group in both cell lines exhibited stronger migration ability than the model group, indicating that glycyrrhizin A can protect cell migration ability, play an important role in burns, and promote wound healing.

[0081] Performance Test 17: Effects of hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 on cellular oxidative stress indices LDH, GSH-PX, and CAT are all important indicators reflecting the level of oxidative stress. This experiment evaluated the protective effect of LCA by detecting various oxidative indicators in two cell lines. The detection results of Example 1 are as follows: Figures 28-29 As shown, the detection results of other embodiments are shown in the figure. Figures 28-29 Basically the same, such as Figure 28 As shown, in L929 cells, the LCA group had significantly higher levels of GSH-PX and CAT compared to the model group, while the model group had significantly higher levels of LDH than both the LCA group and the control group. In A9 cells, the experimental results were consistent with those in L929 cells. Figure 29As shown, the levels of GSH-PX and CAT in the LCA group were significantly higher than those in the model group (P<0.05), while the LDH level was higher in the LCA group and the control group. This indicates that LCA can increase the levels of GSH-PX and CAT, enabling cells to retain their antioxidant capacity under hydrogen peroxide stimulation.

[0082] Performance Test 18: The hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 inhibited inflammatory factors in cells. IL-6, IL-1β, and TNF-α are all pro-inflammatory cytokines and major inflammatory cytokines mediating the progression of burns. They each play different roles during inflammation in the body. Therefore, this study used enzyme-linked immunosorbent assay (ELISA) to determine the effect of glycyrrhizin chalcone A on the levels of three inflammatory factors (IL-6, IL-1β, and TNF-α) in H2O2-induced fibroblasts. The detection results in Example 1 are as follows: Figures 30-31 As shown, the detection results of other embodiments are shown in the figure. Figures 30-31 Basically the same, the results are as follows Figures 30-31 As shown, the levels of the three inflammatory factors in the blank group cells were significantly lower than those in the model group (P<0.001). Compared with the model group, the LCA group also significantly reduced the levels of IL-6, IL-1β, and TNF-α in cells after H2O2 induction (P<0.01, P<0.05, P<0.05), indicating that LCA has an antagonistic effect on the increase of inflammatory factor levels in vascular endothelial cells after H2O2 induction.

[0083] Performance Test 18: Effects of the hydrogel dressings prepared in Examples 1-7 and Comparative Examples 1-3 on organelles in cells. Transmission electron microscopy (TEM) is a powerful tool capable of observing the ultrastructure of cells and tissues at extremely high resolution. Compared to optical microscopy, TEM uses electron beams instead of light for imaging, thus revealing details of nanoscale structures such as organelles, protein complexes, and cell membranes. This experiment aims to use TEM technology to observe the ultrastructure of cells and gain insights into morphological changes in cells under drug preprotection and hydrogen peroxide stimulation conditions. The detection results of Example 1 are as follows: Figure 32 As shown, the detection results of other embodiments are shown in the figure. Figure 32 Basically the same, such as Figure 32 As shown, the mitochondrial structure of the blank group cells was intact, with clearly visible cristae. The mitochondria in the LCA group were also relatively intact, with clearly visible cristae. However, the mitochondrial structure of the model group was incomplete, and the mitochondria were fragmented. This demonstrates that glycyrrhizin A has a protective effect against hydrogen peroxide-induced cell damage.

[0084] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be covered within the scope of the claims and specification of the present invention.

Claims

1. A glycyrrhizin chalcone A gel cream with anti-inflammatory and analgesic functions, characterized in that, By weight percentage, it includes 0.5%–1.5% carbomer 940, 5%–10% glycerol, 1%–2% pH adjuster, 87%–92% glycyrrhizin A, and the balance being water.

2. The glycyrrhizin chalcone A gel ointment with anti-inflammatory and analgesic functions according to claim 1, characterized in that, The pH value of the licorice chalcone A gel is 6.5~7.

5.

3. The glycyrrhizin chalcone A gel ointment with anti-inflammatory and analgesic functions according to claim 2, characterized in that, The pH value of the glycyrrhizin chalcone A gel is 7.

4. The glycyrrhizin chalcone A gel ointment with anti-inflammatory and analgesic functions according to claim 1, characterized in that, The pH adjuster includes triethanolamine.

5. A method for preparing a glycyrrhizin chalcone A gel ointment with anti-inflammatory and analgesic functions, characterized in that, Includes the following steps: Dissolve Carbomer 940 in water; Add glycerin and a pH adjuster to adjust viscosity and pH value; After dissolving in glycyrrhizin A, dilute to volume with water.

6. The method for preparing a glycyrrhizin chalcone A gel ointment with anti-inflammatory and analgesic functions according to claim 5, characterized in that, Adjust the viscosity to 130 mPa.s~190 mPa.s.

7. The method for preparing a glycyrrhizin chalcone A gel ointment with anti-inflammatory and analgesic functions according to claim 6, characterized in that, The viscosity was adjusted to 140 mPa·s.

8. A medicine for treating burns, characterized in that, This includes the glycyrrhizin chalcone A gel as described in any one of claims 1 to 4, or the glycyrrhizin chalcone A gel prepared by the preparation method described in any one of claims 5 to 7.

9. A gel plaster, characterized in that, This includes the glycyrrhizin chalcone A gel as described in any one of claims 1 to 4, or the glycyrrhizin chalcone A gel prepared by the preparation method described in any one of claims 5 to 7.

10. A traditional Chinese medicine patch, characterized in that, Includes the gel plaster as described in claim 9.

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