An antibacterial anti-inflammatory acne-removing and acne and folliculitis improving composition and use thereof

Abstract

The application discloses an antibacterial, anti-inflammatory, acne-removing, acne and folliculitis improving composition and application thereof. The composition provided by the application contains the following components in a mass ratio: nano-biological sulfur:salicylic acid:gentian root extract:trans-4, 5- dihydroxy-4-(2-hydroxyethyl)-2-methyl-2H-furan-3-one:succinic acid=(1-5):(5000-50000):(5000-50000):(10000-50000):(2000-10000). The composition has significant antibacterial and anti-inflammatory effects, and the components have synergistic effects. The composition can destroy pathogenic bacterial cell membranes, inhibit staphylococcus aureus, propionibacterium acnes and malassezia, reduce the level of inflammatory factors, remove acne, improve acne and folliculitis, relieve skin discomfort, make the skin look fresh, and be used for preparing more skin disease treatment drugs and acne-removing products.

Description

Technical Field

[0001] This invention relates to the fields of pharmaceutical and cosmetic technology, and more specifically, to a composition for antibacterial, anti-inflammatory, acne-reducing, and folliculitis-improving purposes, and its application. Background Technology

[0002] In recent years, due to factors such as increased life stress and changes in dietary habits, the incidence of inflammatory skin diseases such as acne and folliculitis has been rising year by year. These diseases not only affect the appearance of patients but may also lead to mental health problems and reduce their quality of life. Therefore, in-depth research into the pathogenesis of acne and folliculitis and the search for safer and more effective treatments have become a hot topic and focus of medical and cosmetic research, aiming to provide patients with better treatment options and improve their skin condition and quality of life.

[0003] Existing acne and folliculitis treatment products commonly suffer from significant side effects and limited functionality (only inhibiting bacteria or only repairing skin). For example, topical retinoids such as isotretinoin gel and adapalene are highly irritating (causing erythema and desquamation), require sun protection, are contraindicated in pregnant women, and have a slow onset of action (4-8 weeks). Topical antibacterial medications such as benzoyl peroxide can cause dry skin or allergies. Oral antibiotics such as doxycycline and minocycline can disrupt the gut microbiota, leading to drug resistance (e.g., Propionibacterium acnes shows a resistance rate of over 50% to erythromycin), thus affecting efficacy.

[0004] Sulfur, a yellow non-metallic element widely distributed in the Earth's crust, is a long-established and effective medicine for treating various skin diseases. Topical application of sulfur has insecticidal, antibacterial, antipruritic, moisturizing, anti-inflammatory, keratinocyte metabolism-promoting, and sebum-regulating effects. Colloidal sulfur is a preparation in which fine sulfur particles are dispersed in water or oil phases. Listed by the FDA as an OTC anti-acne ingredient, it has long been used to treat acne, seborrheic dermatitis, rosacea, and other skin inflammations due to its antibacterial, anti-inflammatory, and oil-controlling properties. However, traditional colloidal sulfur particles are typically micron-sized, making it difficult to penetrate deep into hair follicles, thus limiting its antibacterial effect. During storage, sulfur particles tend to agglomerate, reducing dispersibility and bioavailability. Furthermore, coarse sulfur particles can easily cause dry skin and peeling; sensitive skin may experience erythema or burning sensations after use, leading to skin sensitivity and allergic reactions. Long-term use can also irritate the skin, causing itching, redness, and other discomfort. In severe cases, it can lead to skin erosion, infection, and damage to the skin barrier, resulting in pigmentation. Therefore, existing dermatological products containing sulfur for the treatment of acne or folliculitis still suffer from problems such as low bioavailability, limited antibacterial effect, significant side effects, and limited functionality (only inhibiting bacteria, unable to repair). Summary of the Invention

[0005] The technical problem to be solved by the present invention is to overcome the problems of large side effects, single function (only antibacterial) and limited antibacterial effect that are common in existing sulfur-containing acne and folliculitis treatment products. The present invention provides a composition that is antibacterial, anti-inflammatory and acne-removing and improves acne and folliculitis and its application.

[0006] The first objective of this invention is to provide an antibacterial, anti-inflammatory, and acne-reducing composition.

[0007] A second object of the present invention is to provide a method for preparing the composition.

[0008] A third object of the present invention is to provide applications of the composition.

[0009] The fourth objective of this invention is to provide a product.

[0010] A fifth object of the present invention is to provide applications of the above-described compositions or products.

[0011] The above-mentioned objective of this invention is achieved through the following technical solution:

[0012] This invention provides an antibacterial, anti-inflammatory, and acne-removing composition, wherein the composition contains the following components in the following mass ratio: nano-biological sulfur: salicylic acid: gentian root extract: tranexamic acid: succinic acid = (1-5): (5000-50000): (5000-50000): (10000-50000): (2000-10000).

[0013] This invention provides a multi-target synergistic, mild, and highly effective antibacterial, anti-inflammatory, and acne-reducing composition. Through the synergistic combination of nano-biosulfur and salicylic acid, along with the multiple mechanisms of action of natural extracts, tranexamic acid, and succinic acid, it exhibits a synergistic effect, achieving targeted inhibition of acne and folliculitis pathogens. Simultaneously, it regulates sebum secretion, reduces inflammation, and repairs the skin barrier, achieving root-cause acne treatment and skin rejuvenation. The composition provided by this invention overcomes the limitations of existing sulfur-containing acne and folliculitis treatment products, such as limited antibacterial effects, significant side effects, and single-function limitations. It effectively acts on the cell membrane of pathogenic bacteria, significantly enhancing bactericidal and anti-inflammatory effects. It is gentle and non-irritating, has a repairing effect on the skin, and can improve acne and folliculitis. It can be used to prepare more drugs for skin diseases and acne-reducing products.

[0014] Preferably, the composition contains the following components in the following mass ratio: nano-biosulfur: salicylic acid: gentian root extract: tranexamic acid: succinic acid = (1-3): (5000-10000): (5000-10000): (10000-20000): (2000-10000).

[0015] The nano-active sulfur used in this invention is a nano-active substance with a particle size of less than 100 nm, developed using a nano-delivery system (NDS). Its extremely small particle size and large specific area result in better efficacy. Targeting inflammation caused by Propionibacterium acnes infection and hair follicle blockage, nano-active sulfur can directly act on the bacteria and blocked hair follicles, increasing the local effective concentration and enhancing its bactericidal and anti-inflammatory effects. This invention, through compounding with other components to form a stable formulation, maintains its efficacy while reducing the dosage or concentration of nano-active sulfur, improving its effectiveness, and avoiding the side effects associated with sulfur-containing products.

[0016] Preferably, the particle size of the nano-biosulfur is 20-80 nm.

[0017] Preferably, the gentian root extract in the composition is prepared by extracting the gentian rhizome (Gentiana scabra) using conventional methods in the art through soaking in an aqueous ethanol solution.

[0018] More preferably, the preparation method of the gentian root extract is as follows: after crushing and sieving the gentian rhizome, it is mixed with 50-80% ethanol aqueous solution at a material-to-liquid ratio of 1:8-12 mg / L, and then extracted at a temperature of 40-55℃ for 2-3 hours; after extraction, the mixture is filtered, the filtrate is collected, and the gentian root extract is obtained.

[0019] The present invention provides a method for preparing the above composition, comprising the following steps:

[0020] S1. Weigh out nano-biological sulfur, salicylic acid, gentian root extract, tranexamic acid, and succinic acid according to the specified proportions.

[0021] S2. Add the above raw materials to the solvent, mix at 25-30°C, and stir at 200-300 r / min for 20-30 minutes until completely homogenized to obtain the composition.

[0022] Preferably, the solvent is selected from hydrophilic solvents such as pure water and phosphate buffered solution (PBS).

[0023] This invention provides the application of the above composition in the preparation of antibacterial, anti-inflammatory, and acne-removing products.

[0024] The present invention provides the use of the above composition in the preparation of dermatological drugs for treating acne or folliculitis.

[0025] This invention provides a product containing an antibacterial, anti-inflammatory, and acne-removing composition.

[0026] Preferably, the product contains the following components in weight percentage: 0.0000125-0.1% nano-biosulfur, 0.1-2% salicylic acid, 0.1-1% gentian root extract, 0.1-1% tranexamic acid, and 0.05-0.2% succinic acid; the product also contains a solvent.

[0027] More preferably, the product contains the following components in weight percentage: 0.0000125-0.01% nano-biosulfur, 0.1-0.5% salicylic acid, 0.1-0.5% gentian root extract, 0.1-1% tranexamic acid, and 0.05-0.2% succinic acid.

[0028] Specifically, this invention provides a product containing an antibacterial, anti-inflammatory, and acne-reducing composition. The product, after formulation and use, significantly reduces the concentration of nano-biosulfur (0.0000125-0.1%), which is extremely low compared to the 1-3% concentration used in commercially available sulfur-containing products. Furthermore, this invention maintains a high level of antibacterial and anti-inflammatory effects while reducing the concentration of nano-biosulfur, and works synergistically with other components to regulate sebum secretion and repair the skin barrier. The composition is more effective than commercially available sulfur-containing products, is gentler, more efficient, and non-irritating, and also has a repairing effect.

[0029] Preferably, the product is a gel, cream, serum, mask, or spray.

[0030] The present invention also provides the use of the above-described composition or product in acne treatment.

[0031] The present invention has the following beneficial effects:

[0032] This invention provides a multi-target synergistic, mild, and highly effective antibacterial, anti-inflammatory, and acne-removing composition. The composition utilizes a scientifically and rationally formulated blend of nano-biosulfur, salicylic acid, gentian root extract, tranexamic acid, and succinic acid to regulate the synergistic multiple mechanisms of action of each component. It simultaneously possesses antibacterial, sebum-regulating, inflammatory-reducing, and skin barrier-repairing effects, achieving targeted inhibition of pathogens such as acne and folliculitis. This composition exhibits high inhibitory effects against three skin pathogens: Propionibacterium acnes, Staphylococcus aureus, and Malassezia, demonstrating significant synergistic effects. The prepared product maintains high antibacterial and anti-inflammatory effects while reducing the concentration and dosage of nano-biosulfur, improves hair follicle permeability, and has a repairing effect on the skin. It is mild and highly effective, achieving root-cause acne removal and gentle, highly effective skin rejuvenation, with superior results compared to existing sulfur-containing products. This invention overcomes the problems of existing sulfur-containing acne and folliculitis treatment products, such as large side effects, limited antibacterial effects, and single functions. The provided composition and product can act well on the cell membrane of pathogens, significantly improve antibacterial and anti-inflammatory effects, reduce the concentration and dosage of sulfur, have a repairing effect on the skin, and can improve acne and folliculitis. It can be used to prepare more skin disease drugs and acne removal products. Attached Figure Description

[0033] Figure 1 This is a diagram showing the antibacterial effect of the composition on Propionibacterium acnes.

[0034] Figure 2 This is a diagram showing the antibacterial effect of the composition against Staphylococcus aureus.

[0035] Figure 3 This is a diagram showing the antibacterial effect of the composition against Malassezia.

[0036] Figure 4 This is a scanning electron microscope image of the composition against Propionibacterium acnes.

[0037] Figure 5 This is a scanning electron microscope image of the composition against Staphylococcus aureus.

[0038] Figure 6 This is a scanning electron microscope image of the composition against Malassezia.

[0039] Figure 7 The graph shows the results of membrane potential measurements for different bacterial cell membranes.

[0040] Figure 8 Mg in the supernatant of different bacteria 2+ Concentration statistics results graph.

[0041] Figure 9 Skin phenotypes of mice in different drug-treated models. Detailed Implementation

[0042] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0043] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0044] The gentian root extract used in the examples was prepared by soaking and extracting the gentian rhizome (Gentiana scabra) in an ethanol aqueous solution using conventional methods in the art; commercially available nano-active sulfur with a particle size of 20-80 nm was used.

[0045] Example 1

[0046] A composition comprising the following raw materials in the following mass ratio: salicylic acid: nano-biosulfur: gentian root extract: tranexamic acid: succinic acid = 5000:1:5000:10000:2000.

[0047] The composition is prepared according to the following steps:

[0048] (1) Weigh out salicylic acid, nano-biological sulfur, gentian root extract, tranexamic acid and succinic acid according to the mass ratio;

[0049] (2) The components were diluted with PBS (or pure water or culture medium) to obtain the following concentrations: salicylic acid 0.5%, nano-biological sulfur 0.0001%, gentian root extract 0.5%, tranexamic acid 1%, and succinic acid 0.2%;

[0050] (3) Mix the above components at 25-30℃ and stir at 200-300 r / min for 20-30 minutes until completely homogenized to obtain the final product.

[0051] Example 2

[0052] A composition comprising the following raw materials in the following mass ratio: salicylic acid: nano-biosulfur: gentian root extract: tranexamic acid: succinic acid = 10000:1:10000:20000:4000.

[0053] The preparation method is the same as in Example 1. The concentrations of each component in the prepared composition are as follows: salicylic acid 0.5%, nano-biosulfur 0.00005%, gentian root extract 0.5%, tranexamic acid 1%, succinic acid 0.2%, and the remainder is solvent.

[0054] Example 3

[0055] A composition comprising the following raw materials in the following mass ratio: salicylic acid: nano-biosulfur: gentian root extract: tranexamic acid: succinic acid = 10000:3:10000:20000:4000.

[0056] The preparation method is the same as in Example 1. The concentrations of each component in the prepared composition are as follows: salicylic acid 0.5%, nano-biosulfur 0.00015%, gentian root extract 0.5%, tranexamic acid 1%, succinic acid 0.2%, and the remainder is solvent.

[0057] Comparative Example 1

[0058] A composition comprising the following raw materials in the following mass ratio: salicylic acid: nano-biosulfur = 5000: 1.

[0059] The preparation method is the same as in Example 1. The concentrations of each component in the prepared composition are as follows: salicylic acid 0.5%, nano-biosulfur 0.0001%, and the remainder is solvent.

[0060] Comparative Example 2

[0061] A composition comprising the following raw materials in the following mass ratio: salicylic acid: nano-biosulfur = 10000:1.

[0062] The preparation method is the same as in Example 1. The concentrations of each component in the prepared composition are as follows: salicylic acid 0.5%, nano-biosulfur 0.00005%, and the remainder is solvent.

[0063] Comparative Example 3

[0064] A composition comprising the following raw materials in the following mass ratio: salicylic acid: nano-biosulfur = 10000:3.

[0065] The preparation method is the same as in Example 1. The concentrations of each component in the prepared composition are as follows: salicylic acid 0.5%, nano-biosulfur 0.00015%, and the remainder is solvent.

[0066] Comparative Example 4

[0067] A composition comprising the following raw materials in the following mass ratio: salicylic acid: colloidal sulfur = 5000:1.

[0068] The preparation method is the same as in Example 1. The concentrations of each component in the prepared composition are as follows: salicylic acid 0.5%, colloidal sulfur 0.0001%, and the balance is solvent.

[0069] Comparative Example 5

[0070] A commercially available composition is made from the following raw materials in weight percentages: 2% salicylic acid and 3% sublimed sulfur.

[0071] Comparative Example 6

[0072] A composition comprising the following raw materials in the following mass ratio: salicylic acid: nano-biosulfur: tranexamic acid: succinic acid = 5000:1:10000:1000.

[0073] The preparation method is the same as in Example 1. The concentrations of each component in the prepared composition are as follows: salicylic acid 0.5%, nano sulfur 0.0001%, tranexamic acid 1%, succinic acid 0.1%, and the balance is solvent.

[0074] Comparative Example 7

[0075] A composition comprising the following raw materials in weight percentages: salicylic acid: nano-biosulfur: gentian root extract = 5000: 1: 5000.

[0076] The preparation method is the same as in Example 1. The concentrations of each component in the prepared composition are as follows: salicylic acid 0.5%, nano sulfur 0.0001%, gentian root extract 0.5%, and the remainder is solvent.

[0077] Comparative Example 8

[0078] A composition comprising raw materials in the following mass ratio: nano-biosulfur: succinic acid: tranexamic acid = 1:1000:10000.

[0079] The preparation method is the same as in Example 1. The concentrations of each component in the prepared composition are as follows: nano sulfur 0.0001%, succinic acid 0.1%, tranexamic acid 1%, and the remainder is solvent.

[0080] Test Example 1: Antibacterial Effects of Different Compositions

[0081] The antibacterial effects of different compositions were determined. Compositions of the same concentration and dosage (Examples 1-3, Comparative Examples 1-8) were mixed in liquid agar medium, and their optical density (OD) values ​​were measured using a microplate reader to observe their growth at different concentrations. A commercially available ointment for treating acne folliculitis (containing 1% fusidic acid) was used as a positive control. The microorganism tested was *Propionibacterium acnes* (…). Propionibacterium acnes Staphylococcus aureus ( Staphylococcus aureus Malassezia ( Malassezia spp.).

[0082] The formula for calculating the inhibition rate is: Inhibition rate = Experimental group - Blank control / Negative control - Blank control;

[0083] The results are as follows Figures 1-3As shown in Table 1, the positive control group could not effectively target all three pathogens simultaneously, exhibiting poor inhibitory effects against Malassezia. In Comparative Examples 1-3, the antibacterial effect gradually decreased with decreasing nano-sulfur concentration. In Comparative Example 4, the use of colloidal sulfur instead of nano-biosulfur improved the antibacterial effect, but the improvement was limited, and the use of colloidal sulfur also had significant side effects. The compositions of Examples 1-3, however, showed good inhibitory effects against all three pathogens, maintaining high antibacterial efficacy even after reducing the nano-sulfur concentration. Compared to Comparative Examples 6-8, they demonstrated a significant synergistic effect, showing good inhibitory effects against Staphylococcus aureus, Propionibacterium acnes, and Malassezia. Furthermore, the compositions of the Examples showed better antibacterial effects compared to the commercially available product of Comparative Example 5, which contained a high concentration of sulfur. The bio-nano-sulfur used in the compositions not only had a lower concentration but was also more gentle and efficient, maintaining a high level of antibacterial activity even when combined with other components.

[0084] Table 1. Antibacterial effects of different compositions

[0085]

[0086] Test Example 2: Minimum Inhibitory Concentration (MIC) of Different Compositions

[0087] The compositions based on the examples were evaluated for minimum inhibitory concentration (MIC) using the broth dilution method, in which the compositions were serially diluted in liquid culture medium, and the growth of microorganisms at different concentrations was detected. The antibacterial effect was determined using the same method as in Test Example 1, and the inhibition rate results were statistically analyzed.

[0088] Simultaneously, a negative control group (without the composition) was set up to assess the accuracy and reliability of the experimental results. Multiple concentration gradients of drug or product solutions were prepared to evaluate the dose-dependent antibacterial effects against Staphylococcus aureus, Propionibacterium acnes, and Malassezia.

[0089] (1) Effects on Staphylococcus aureus

[0090] The antibacterial test results of the compositions obtained by serial dilution of different component concentrations are shown in Table 2. The results show that there are significant differences in the antibacterial effect of adjusting the concentration of each component in the composition. The MICs against Staphylococcus aureus are: salicylic acid 0.125%, nano-biological sulfur 0.000025%, gentian root extract 0.125%, tranexamic acid 0.25%, and succinic acid 0.05%.

[0091] Table 2. Inhibitory effects of compositions with different concentrations of components on pathogens.

[0092]

[0093] (2) Effects on Propionibacterium acnes

[0094] The test results are shown in Table 3. The MICs of the composition against Propionibacterium acnes are: salicylic acid 0.125%, nano-biological sulfur 0.000025%, gentian root extract 0.125%, tranexamic acid 0.25%, and succinic acid 0.05%.

[0095] Table 3. Inhibitory effects of compositions with different concentrations of components on pathogens.

[0096]

[0097] (3) Effects on Malassezia

[0098] The test results are shown in Table 4. The MICs of the composition against Propionibacterium acnes are: salicylic acid 0.125%, nano-biological sulfur 0.000025%, gentian root extract 0.125%, tranexamic acid 0.25%, and succinic acid 0.05%.

[0099] Table 4. Inhibitory effects of compositions with different concentrations of components on pathogens.

[0100]

[0101] In summary, the MIC determination of the composition against three pathogens showed significant differences in antibacterial effects after different component concentrations were combined. By using specific concentration ratios, the synergistic effect of each component can be maximized, significantly improving the antibacterial effect. The combination of nano-biosulfur and salicylic acid, along with the multiple mechanisms of action of natural extracts, tranexamic acid, and succinic acid, achieves targeted inhibition of acne and folliculitis pathogens. It can achieve multi-target synergistic, gentle, and highly effective antibacterial and acne-removing effects. It also works synergistically with each component to regulate sebum secretion and repair the skin barrier, achieving the effect of root-cause acne removal and skin rejuvenation.

[0102] Test Example 3: Mechanism of Action of the Composition on the Cell Membrane of Pathogenic Bacteria

[0103] 1. Scanning electron microscopy observation of skin flora

[0104] The colonies of Malassezia, Staphylococcus aureus, and Propionibacterium acnes treated with the compositions of Examples 1-2 and Comparative Example 2 in Test Example 1 were observed by scanning electron microscopy. The colonies of bacteria before treatment were used as a negative control group. The parameters of the electron microscope (such as accelerating voltage, working distance, etc.) were adjusted to observe the morphology of the sample surface.

[0105] Scanning electron microscopy results as follows Figures 4-6As shown, the bacteria in the negative control group (untreated with the composition) exhibited relatively intact and regular morphology. Staphylococcus aureus showed a complete spherical structure, Propionibacterium acnes showed a rod-shaped structure, and Malassezia showed an elliptical structure. However, the bacteria treated with different compositions (Examples 1-2, Comparative Example 2) showed varying degrees of morphological damage. The three bacteria in the negative control group had smooth, plump surfaces without obvious pits or cracks. When treated with the compositions of Examples 1-2, the bacterial morphology was significantly damaged, and some bacteria even completely adhered together. The results demonstrate the irreversible damage to bacterial morphology caused by the compositions. As shown in the figure, Examples 1 and 2 caused greater damage to the three microbial communities than Comparative Example 2.

[0106] 2. Measurement of bacterial cell membrane potential

[0107] The cell membrane potential of each pathogen after the above treatment was measured to understand its physiological state and metabolic activity. Different bacterial cells were labeled with fluorescent probes (such as DiBAC4(3)) and the changes in cell membrane potential were measured by a fluorescence spectrophotometer. The DiBAC4(3) molecular probe can bind to proteins in the cytoplasm inside the cell. By changing its luminescence intensity, it can be determined whether the cell membrane is damaged. When the cell membrane potential becomes more positive (depolarization), the intracellular potential is relatively more positive, and DiBAC4(3) will enter the cell more, resulting in an increase in fluorescence intensity. Conversely, when the cell membrane potential becomes more negative (depolarization), DiBAC4(3) will be more distributed outside the cell, resulting in a decrease in fluorescence intensity.

[0108] The measurement results are as follows Figure 7 As shown in Table 5, the average fluorescence intensity after treatment with the compositions of the examples and comparative examples was lower than that of the normal control group, indicating that the compositions can cause depolarization of the cell membranes of the three microorganisms. This can inhibit bacterial cell growth and reproduction by inhibiting energy metabolism, affecting ion balance, and disrupting cell membrane integrity. The fluorescence intensity of Examples 1 and 2 was even lower, which caused hyperpolarization of Staphylococcus aureus and inhibited bacterial cell growth and reproduction.

[0109] Table 5 Results of bacterial cell membrane potential measurement

[0110]

[0111] 3. Bacterial cell membrane permeability assay

[0112] When bacteria are damaged, the permeability of the bacterial cell membrane changes, causing a large number of important substances, such as magnesium, to leak out. 2+ K + Proteins and other substances leak out of the bacteria, inhibiting their growth and reproduction. Further, the Mg in the supernatant of the treated bacteria is then analyzed.2+ The concentration was measured.

[0113] The measurement results are as follows Figure 8 As shown in Table 6, the Mg content in the supernatant of the tested Staphylococcus aureus and Propionibacterium acnes is... 2+ The concentration was relatively high, while the supernatant containing Mg in the test bacteria Malassezia was relatively high. 2+ The concentration was relatively low. The Mg content in the supernatant of Staphylococcus aureus treated with the three compositions of Comparative Example 2, Example 1, and Example 2 was low. 2+ The concentrations were 0.1220, 0.2292, and 0.1509 mmol / L, respectively, all showing significant increases. Furthermore, statistically significant differences were observed in the Mg concentrations of Examples 1 and 2 compared to the negative control group (untreated). 2+ The concentration was significantly higher than that of Comparative Example 2, and it exerted an antibacterial effect by disrupting the cell membrane of Staphylococcus aureus. Similarly, after co-culturing with Propionibacterium acnes overnight, the Mg content in the supernatant of Comparative Example 2, Example 1, and Example 2 was significantly higher. 2+ The concentrations were 0.1430, 0.1635, and 0.1548 mmol / L, respectively, all showing significant increases compared to the negative control group, demonstrating statistically significant differences. Furthermore, the Mg concentrations in the supernatants of Examples 1 and 2 were significantly higher. 2+ The concentrations were all greater than those of Comparative Example 2.

[0114] Mg in the supernatant after co-culturing three compositions with Malassezia overnight 2+ The concentrations were 0.01038, 0.01028, and 0.01314 mmol / L, respectively, all of which showed an increase compared to the negative control group, although no significant difference was observed between the groups. p >0.05), but the compositions in the examples and comparative examples all increased Mg. 2+ The concentration ultimately inhibits bacterial growth and reproduction.

[0115] Table 6. Mg in supernatants of different bacteria 2+ Concentration results

[0116]

[0117] Test Example 4: Determination of Anti-inflammatory Effect and Cell Safety

[0118] 1. Cytotoxicity test MTT assay

[0119] The seeded Hacat cells were placed in 96-well plates at a density of 8 × 10⁶ cells per well. 3Cells were divided into a control group and six treatment groups. The control group was cultured in DMEM. After incubating the 96-well plates in a constant temperature incubator for 24 hours, the culture medium was discarded. The prepared compositions of Example 1 (high concentration) and Example 2 (low concentration) were diluted halfway with DMEM to six concentration gradients and added to the six treatment groups. The control group was added with DMEM. The plates were then incubated in an incubator for 24 hours. Subsequently, the cell supernatant was removed, and the cells were washed 1-2 times with PBS. In the dark, 100 μL of 0.5 mg / mL MTT in DMEM solution was added to each well. After incubating the cell culture plates at 37°C for 4 hours, the MTT solution was removed. The cell supernatant was removed, and 150 μL of DMSO was added to each well. After shaking evenly, the absorbance of each well was estimated at 570 nm using a microplate analyzer.

[0120] The results are shown in Table 7. The experimental results indicate that the low concentration diluted to 1 / 8 and the high concentration diluted to 1 / 16 of the composition were not toxic to cells and had good safety.

[0121] Table 7 Cytotoxicity Results

[0122]

[0123] 2. Keratinocyte inflammatory factor release experiment

[0124] Keratinocytes were seeded at a density of 200,000 cells / mL, with 2 mL per well. In the blank control group, a solution containing only culture medium was added. LPS was added to both the model group and the test substance group to induce the release of inflammatory factors from keratinocytes. The test substance groups were each added with the same amount of the test substance (high concentration group - Example 1; low concentration group - Example 2), at a concentration equal to 1 / 16 of the original solution from the examples. Following the ELISA kit instructions, the concentrations of inflammatory factors (e.g., IL-1β, IL-6, IL-8) in the supernatant of each well were measured.

[0125] The results are shown in Table 8. It can be seen that the content of inflammatory factors in the model group was significantly increased. The treatment with the composition of the example can inhibit the expression of LPS-induced inflammatory factors IL-1β, IL-6 and IL-8, and has a good anti-inflammatory effect. It can reduce skin inflammation and be better applied to the preparation of drugs for inflammatory skin diseases.

[0126] Table 8. Expression results of inflammatory factors IL-1β, IL-6, and IL-8

[0127]

[0128] Test Example 5: Therapeutic effect of the composition on acne and folliculitis

[0129] Animal models of Malassezia and Propionibacterium acnes were established. In each group, acne (left side of mouse back) / folliculitis (right side of mouse back) models were established by topical application of 100% oleic acid (0.05 mL / mouse) to the back skin combined with subcutaneous injection of Propionibacterium acnes / Malassezia bacterial solution (0.1 mL / mouse). Typical acne-like skin lesions such as desquamation, erythema, and dilated pores were induced. The drug treatment groups included a high-concentration group (Example 1) and a low-concentration group (Example 2). A positive control group was also established, treating Malassezia with terbinafine spray (antifungal) and Propionibacterium acnes with fusidic acid (antibacterial). Specific treatment methods are shown in Table 9.

[0130] Table 9 Different experimental treatment methods

[0131]

[0132] Based on the above treatment, the skin of mice in each group was observed, and the skin condition of each group of mice was scored according to the binary evaluation criteria summarized by SCORAD and Kong Yue et al., as shown in Table 10.

[0133] Table 10 Binary Evaluation Criteria

[0134]

[0135] The results are as follows Figure 9 As shown in Table 11, the model group exhibited significantly more redness, swelling, and damage, with noticeable crusting, compared to the control group. After treatment with the composition, the redness and damage were significantly improved, resulting in smoother, more delicate skin with reduced crusting. The high-concentration group showed even better results, demonstrating a significant skin-repairing effect. This is attributed to the synergistic antibacterial, anti-inflammatory, and repairing effects of the components in the composition. Gentian root extract possesses anti-inflammatory and soothing properties, effectively alleviating skin discomfort; tranexamic acid stabilizes the skin condition; and succinic acid has antioxidant and anti-inflammatory effects, further promoting skin repair. The composition provided by this invention reduces the potential irritation risks associated with salicylic acid alone and salicylic acid + sulfur preparations, while effectively repairing skin damage caused by pathogens. It maintains healthy and stable skin while exerting antibacterial and anti-inflammatory effects, helping the skin to rejuvenate.

[0136] Table 11 Evaluation Results (aq2)

[0137]

[0138] The above embodiments of the present invention are merely examples for clearly illustrating the present invention and are not intended to limit the implementation of the present invention. Those skilled in the art can make other variations or modifications based on the above description. It is neither necessary nor possible to exhaustively describe all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. An antibacterial, anti-inflammatory, acne-removing composition, characterized in that, The composition is composed of components in the following mass ratio: nanobiological sulfur: salicylic acid: gentian root extract: tranexamic acid: succinic acid = (1-5): (5000-50000): (5000-50000): (10000-50000): (2000-10000).

2. The composition of claim 1, wherein, The composition is composed of components in the following mass ratio: nanobiological sulfur: salicylic acid: gentian root extract: tranexamic acid: succinic acid = (1-3): (5000-10000): (5000-10000): (10000-20000): (2000-10000).

3. A method for preparing the composition of claim 1 or 2, comprising the following steps: S1. Nanobiological sulfur, salicylic acid, gentian root extract, tranexamic acid, and succinic acid are weighed in proportion respectively; S2. The above raw materials are added to a solvent, mixed at 25-30°C, and stirred at 200-300 r / min for 20-30 minutes until completely homogenized to obtain the composition.

4. The preparation method according to claim 3, characterized in that, The solvent is pure water or a phosphate buffer solution.

5. Use of the composition of claim 1 or 2 in the preparation of antibacterial and anti-inflammatory drugs.

6. Use of the composition of claim 1 or 2 in the preparation of acne-removing products.

7. Use of the composition of claim 1 or 2 in the preparation of dermatological drugs for treating acne or folliculitis.

8. A cosmetic product, characterized by, The composition of claim 1 or 2 is contained.

9. The cosmetic product of claim 8, wherein, The cosmetic also contains a solvent.

10. The cosmetic product of claim 9, wherein The cosmetic is a gel, cream, serum, mask, or spray.

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