An external composition for improving acne and use thereof

Abstract

This invention relates to the field of biomedical technology, specifically to a topical composition for improving acne and its application. The topical composition is made from pharmaceutical raw materials including a first component, a second component, and a third component. The first component comprises 25-35 parts of *Ampelopsis japonica*, 25-35 parts of hawthorn, 25-35 parts of *Salvia miltiorrhiza*, 13-17 parts of gardenia, and 8-12 parts of *Bletilla striata*. The second component comprises 25-35 parts of mother-of-pearl, 13-17 parts of tribulus terrestris, and 8-12 parts of safflower. The third component comprises 13-17 parts of raw rhubarb, 8-12 parts of gallnut, 8-12 parts of turmeric, 3-7 parts of *Ganoderma lucidum*, and 3-7 parts of asparagus. This invention also provides a topical composition consisting of the first, second, or third component alone. The topical composition of this invention has dual effects of inhibiting *Propionibacterium acnes* and anti-inflammatory properties, and can be further prepared into cosmetics or pharmaceuticals for the prevention, relief, and treatment of various types of acne and the improvement of acne scars. Clinical trials have shown significant effects, high safety, and promising application prospects.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a topical composition for improving acne and its application. Background Technology

[0002] Acne is a chronic inflammatory skin disease of the pilosebaceous unit, commonly occurring on the face, back, and chest—areas rich in sebaceous glands. Its clinical manifestations are diverse, including comedones (blackheads, whiteheads), inflammatory papules, pustules, nodules, and cysts. Severe cases can leave atrophic or hypertrophic scars and pigmentation, significantly impacting the patient's appearance and mental health. Statistics show that approximately 80% of adolescents experience acne to varying degrees, and some middle-aged men and women are also troubled by it.

[0003] Modern medicine believes that the pathogenesis of acne is related to multiple factors, mainly including: elevated androgen levels leading to excessive sebum secretion, abnormal keratinization of the pilosebaceous duct, microbial infection such as Propionibacterium acnes, and the resulting inflammatory response and immune dysregulation. Currently, commonly used clinical treatments include topical retinoids, benzoyl peroxide, and antibiotics, as well as oral antibiotics, isotretinoin, and anti-androgens. However, these therapies generally suffer from problems such as skin irritation, drug resistance, high relapse rates after discontinuation, and poor patient compliance (see CN114832065A).

[0004] Traditional Chinese medicine (TCM) has a long history of treating acne, emphasizing syndrome differentiation and treatment, and addressing both internal and external factors. It possesses unique advantages such as multi-target therapy, fewer side effects, and lower risk of drug resistance. Chinese patent CN114832065A discloses a TCM composition for treating acne, consisting of Salvia miltiorrhiza, Coptis chinensis, Angelica dahurica, Cynanchum paniculatum, Bletilla striata, Astragalus membranaceus, Trichosanthes kirilowii, Taraxacum mongolicum, and Lonicera japonica, formulated as a topical gel with a clinical overall effective rate of 93.3%. Chinese patent CN110201093A discloses an acne gel composed of Scutellaria baicalensis, Coptis chinensis, Phellodendron chinense, Sophora flavescens, Angelica dahurica, Bletilla striata, Poria cocos, and Salvia miltiorrhiza, used to treat various types of acne and surgical surface infections, exhibiting anti-inflammatory and swelling-reducing effects. Chinese patent CN110201128B discloses a traditional Chinese medicine preparation for treating acne caused by phlegm and blood stasis, composed of seventeen herbs including turmeric, coix seed, bupleurum, cyperus, and tangerine peel, utilizing their effects of soothing the liver and regulating qi, strengthening the spleen and eliminating dampness, promoting blood circulation and removing blood stasis, and softening and dispersing nodules. Chinese patent CN108543044B provides a combined internal and external treatment drug; the internal formula warms yang and resolves dampness, while the external formula dispels wind, clears heat, detoxifies, and disperses nodules. Clinical trials show that it can significantly improve acne symptoms. These patents provide valuable insights for the treatment of acne with traditional Chinese medicine from different perspectives.

[0005] However, existing traditional Chinese medicine compound formulas often have complex components, and the scientific implications of their compatibility and the interaction mechanisms between components are still unclear, lacking support from modern pharmacological data. Therefore, developing a topical composition for improving acne with clear components, scientific compatibility, and a well-defined mechanism of action has significant clinical and market value. Summary of the Invention

[0006] To address the aforementioned technical problems, the present invention aims to provide a topical composition for improving acne and its application. The topical composition has dual effects of inhibiting Propionibacterium acnes and anti-inflammation. In particular, the whole formula exhibits a significant synergistic effect in anti-inflammatory activity in vivo. It can be prepared into cosmetics or topical medicines for the prevention, relief and treatment of various types of acne and the improvement of acne marks and scars.

[0007] To achieve the above-mentioned technical effects, the present invention adopts the following technical solution: In a first aspect, the present invention provides a topical composition for improving acne, said topical composition being made from the following pharmaceutical ingredients: Group 1: 25-35 parts of Ampelopsis japonica, 25-35 parts of Crataegus pinnatifida, 25-35 parts of Salvia miltiorrhiza, 13-17 parts of Gardenia jasminoides, and 8-12 parts of Bletilla striata; Second component: 25-35 parts mother-of-pearl, 13-17 parts tribulus terrestris, 8-12 parts safflower; The third component consists of 13-17 parts of raw rhubarb, 8-12 parts of gallnut, 8-12 parts of turmeric, 3-7 parts of Ganoderma lucidum, and 3-7 parts of asparagus.

[0008] As a further preferred technical solution, the first component is made from the following medicinal raw materials: 30 parts of Ampelopsis japonica, 30 parts of Crataegus pinnatifida, 30 parts of Salvia miltiorrhiza, 15 parts of Gardenia jasminoides, and 10 parts of Bletilla striata; the second component is made from the following medicinal raw materials: 30 parts of pearl powder, 15 parts of Tribulus terrestris, and 10 parts of Carthamus tinctorius; the third component is made from the following medicinal raw materials: 15 parts of raw rhubarb, 10 parts of Galla chinensis, 10 parts of Curcuma longa, 5 parts of Ganoderma lucidum, and 5 parts of Asparagus cochinchinensis.

[0009] In a second aspect, the present invention provides a topical composition for improving acne, the topical composition being made from the following pharmaceutical raw materials: 25-35 parts of Ampelopsis japonica, 25-35 parts of Crataegus pinnatifida, 25-35 parts of Salvia miltiorrhiza, 13-17 parts of Gardenia jasminoides, and 8-12 parts of Bletilla striata.

[0010] As one of the preferred technical solutions, the external composition is made from the following pharmaceutical raw materials: 30 parts of Ampelopsis japonica, 30 parts of Crataegus pinnatifida, 30 parts of Salvia miltiorrhiza, 15 parts of Gardenia jasminoides, and 10 parts of Bletilla striata.

[0011] Thirdly, the present invention provides a topical composition for improving acne, the topical composition being made from the following pharmaceutical raw materials: 25-35 parts of mother-of-pearl, 13-17 parts of tribulus terrestris, and 8-12 parts of safflower.

[0012] As one of the preferred technical solutions, the external composition is made from the following pharmaceutical raw materials: 30 parts of mother-of-pearl, 15 parts of tribulus terrestris, and 10 parts of safflower.

[0013] Fourthly, the present invention provides a topical composition for improving acne, the topical composition being made from the following pharmaceutical raw materials: 13-17 parts of raw rhubarb, 8-12 parts of gallnut, 8-12 parts of turmeric, 3-7 parts of Ganoderma lucidum, and 3-7 parts of asparagus.

[0014] As one of the preferred technical solutions, the external composition is made from the following pharmaceutical raw materials: 15 parts of raw rhubarb, 10 parts of gallnut, 10 parts of turmeric, 5 parts of Ganoderma lucidum, and 5 parts of asparagus.

[0015] Fifthly, the present invention provides the use of the topical composition of any one of the first to fourth aspects in the preparation of cosmetics or pharmaceuticals for the prevention, relief and / or treatment of acne.

[0016] As one of the preferred technical solutions, the acne includes one or more of comedones, inflammatory papules, pustules, nodules, and cysts; and / or, the application also includes improving post-acne erythema, post-acne pigmentation, and post-acne scars.

[0017] As one of the preferred technical solutions, the topical composition has the effect of inhibiting the activity of Propionibacterium acnes and / or inhibiting the inflammatory response induced by Propionibacterium acnes.

[0018] In a sixth aspect, the present invention provides a method for preparing the topical composition according to any one of the first to fourth aspects, the method comprising: mixing the pharmaceutical raw materials, extracting with water, and concentrating to obtain an extract of the topical composition.

[0019] As one of the preferred technical solutions, the extraction method includes: mixing the medicinal raw materials, extracting with water three times; adding 5-12 times the amount of water to the medicinal materials for the first extraction and extracting for 0.5-3 hours; adding 5-12 times the amount of water to the medicinal materials for the second extraction and extracting for 0.5-3 hours; adding 5-10 times the amount of water to the medicinal materials for the third extraction and extracting for 0.5-3 hours; separating the extract and residue after each extraction, and adding the residue to the next extraction; combining the three extracts, filtering, and concentrating the filtrate under reduced pressure to a relative density of 1.05-1.10 (measured at 45-55℃) to obtain the extract (extract paste) of the external composition.

[0020] As a further preferred technical solution, the method further includes the step of spray drying or freeze drying the extract of the topical composition to obtain a dry powder of the extract of the topical composition. The dry powder of the extract is easier to store, quantify, and add to various cosmetic or pharmaceutical bases.

[0021] In a seventh aspect, the present invention provides a cosmetic for improving acne, the cosmetic comprising an extract (extract paste or dried extract powder thereof) of the topical composition as described in any one of the first to fourth aspects as an active ingredient, the cosmetic being prepared as a serum, gel, lotion, cream, mask, lotion, spray or gel.

[0022] As one of the preferred technical solutions, the extract, calculated as extract powder, accounts for 0.5%-30% of the total weight of the cosmetic. For example, for daily care products, 0.5%-5% is preferred; for therapeutic products, 5%-30% is preferred. When using extract paste, its addition amount is calculated based on the dry matter content therein to the above range.

[0023] Eighthly, the present invention provides a medicament for improving acne, the medicament comprising an extract (extract paste or dried extract powder thereof) of a topical composition as described in any one of the first to fourth aspects as an active ingredient, and pharmaceutically acceptable excipients.

[0024] As one of the preferred technical solutions, the extract, calculated as dry powder, accounts for 1%-30% of the total weight of the drug, depending on the dosage form and therapeutic intensity. When using an extract paste, its addition amount is calculated based on the dry matter content to the above range.

[0025] The medicine can be formulated into topical dosage forms such as ointments, creams, gels, lotions, liniments, pastes, patches, films, or sprays. These dosage forms are convenient for direct application to acne-affected areas, exerting the effects of inhibiting Propionibacterium acnes, reducing inflammation, and improving acne scars.

[0026] Compared with the prior art, the present invention has the following beneficial effects: First, the topical composition of this invention is scientifically formulated, with significant synergistic effects among its components. In in vivo anti-inflammatory experiments on zebrafish at the same total drug concentration, the acne-reducing efficacy of the complete formula was not only significantly higher than that of any single component used alone, but also far higher than that of any two components used in combination. This result fully demonstrates that the three-component combination is not a simple additive effect, but rather produces an unexpected synergistic effect in inhibiting the inflammatory response induced by Propionibacterium acnes, reflecting the scientific nature and innovation of the formulation of this invention.

[0027] Secondly, the topical composition of this invention possesses both inhibitory and anti-inflammatory effects on Propionibacterium acnes, offering comprehensive efficacy and high safety. In vitro antibacterial experiments showed that, at the same drug concentration, the overall formulation exhibited a 99.89% inhibition rate against Propionibacterium acnes, with a surviving count of only 40 CFU / mL, significantly superior to the single-use groups and the combined two-component groups. In vivo anti-inflammatory experiments in zebrafish showed that the overall formulation significantly reduced neutrophil infiltration into the yolk sac inflamed areas, rapidly alleviating inflammation. Furthermore, human trials demonstrated that no adverse reactions such as skin irritation or allergies occurred after 10 days of continuous use, indicating extremely high safety. Comparisons of typical cases before and after treatment showed rapid onset of action and good patient compliance.

[0028] Finally, this invention has wide applications and can improve various acne-related symptoms. Human efficacy evaluation results show that the skin lesion efficacy index of the full-treatment group reached 71.2%, with a significant improvement rate as high as 89.5%. This topical composition not only effectively eliminates acne lesions such as pimples, inflammatory papules, pustules, nodules, and cysts, but also simultaneously improves residual red acne marks, pigmentation, and early scars, comprehensively enhancing skin health. Furthermore, this composition can be prepared into various dosage forms such as serums, gels, lotions, creams, masks, toners, sprays, or gels to meet the usage habits and needs of different patients, demonstrating promising market application prospects. Attached Figure Description

[0029] Figure 1 This is a typical fluorescence micrograph of neutrophil aggregation in a zebrafish local inflammation model; Figure 2 These are before-and-after photos of typical case 1 (facial acne) in Embodiment 3 of the present invention; Figure 3 These are before-and-after photos of typical case 2 (mandibular acne) in Embodiment 3 of the present invention. Detailed Implementation

[0030] The embodiments of the technical solution of the present invention will now be described in detail with reference to the accompanying drawings. The following embodiments are only used to more clearly illustrate the technical solution of the present invention, and are therefore merely examples and should not be used to limit the scope of protection of the present invention. Those skilled in the art should understand that the present invention can be implemented even without certain specific details. In some other embodiments, methods, means, equipment, and steps well known to those skilled in the art are not described in detail in order to highlight the main points of the present invention. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Unless otherwise specified, all units used in this specification are International Standard Units (SI), and the numerical values ​​and ranges appearing in this invention should be understood to include systematic errors unavoidable in industrial production.

[0031] All Chinese medicinal materials used in this embodiment were purchased from commercially available products that meet the standards of the Chinese Pharmacopoeia, and their pharmacopoeia basis is as follows: White peony root: the dried tuberous root of Ampelopsis japonica (Thunb.) Makino, a plant in the Vitaceae family.

[0032] Hawthorn: The dried, ripe fruit of Crataegus pinnatifida Bge. var. major NEBr. or Crataegus pinnatifida Bge., both belonging to the Rosaceae family.

[0033] Danshen: The dried root and rhizome of Salvia miltiorrhiza Bge., a plant of the Lamiaceae family.

[0034] Gardenia: The dried, ripe fruit of Gardenia jasminoides Ellis, a plant in the Rubiaceae family.

[0035] Bletilla striata: the dried tuber of Bletilla striata (Thunb.) Reichb.f., an orchid.

[0036] Mother-of-pearl: A processed product made by calcining the shells of the three-sailed mussels Hyriopsis cumingii (Lea), Cristariaplicata (Leach), or Pteria martensii (Dunker).

[0037] Tribulus terrestris L., a plant in the family Zygophyllaceae, is the dried, mature fruit of the plant.

[0038] Safflower: The dried tubular flowers of Carthamus tinctorius L., a plant in the Asteraceae family.

[0039] Raw rhubarb: The dried root and rhizome of Rheum palmatum L., Rheum tanguticum Maxim. ex Balf. or Rheum officinale Baill., belonging to the Polygonaceae family, without processing.

[0040] Gallnuts: Galls on the leaves of plants in the Anacardiaceae family, such as Rhus chinensis Mill., Rhus potaninii Maxim., or Rhus punjabensis Stew. var. sinica (Diels) Rehd. et Wils., are mainly formed by the gall aphid Melapis chinensis (Bell) Baker.

[0041] Turmeric: The dried rhizome of Curcuma longa L., a plant in the ginger family.

[0042] Lingzhi: The dried fruiting body of Ganoderma lucidum (Leyss. ex Fr.) Karst. or Ganoderma sinense Zhao, Xu et Zhang, belonging to the Polyporaceae family.

[0043] Asparagus: The dried tuberous root of Asparagus cochinchinensis (Lour.) Merr., a plant in the Liliaceae family. Example 1

[0044] The purpose of this embodiment is to provide a topical composition for improving acne and its preparation method, as detailed below: This topical composition for improving acne is made from the following pharmaceutical ingredients in parts by weight: First group: 30 parts of white peony root, 30 parts of hawthorn, 30 parts of salvia miltiorrhiza, 15 parts of gardenia, and 10 parts of bletilla striata; Second component: 30 parts mother-of-pearl, 15 parts tribulus terrestris, and 10 parts safflower; And the third component: 15 parts raw rhubarb, 10 parts gallnut, 10 parts turmeric, 5 parts Ganoderma lucidum, and 5 parts asparagus.

[0045] The preparation method of this topical composition is as follows: Accurately weigh each medicinal herb slice according to the above formula, mix them evenly, and put them into the extraction tank.

[0046] First extraction: Add 8 times the amount of purified water to the extraction tank, heat to a gentle boil, extract for 1 hour, filter, collect the first extract, and the separated residue enters the second extraction.

[0047] Second extraction: Add 7 times the amount of purified water to the dregs, heat to a gentle boil and extract for 1 hour, filter, collect the second extract, and the separated residue enters the third extraction.

[0048] Third extraction: Add 7 times the amount of purified water to the dregs, heat to a gentle boil and extract for 1 hour, filter, and collect the third extract.

[0049] The three extracts were combined and filtered through a 200-mesh filter cloth. The filtrate was transferred to a vacuum concentration tank and concentrated under reduced pressure at a temperature of 45-55℃ and a vacuum degree of -0.06 to -0.08 MPa until the relative density of the concentrate reached 1.05 to 1.10 (measured at 45-55℃). After spray drying, the extract powder of the external composition of the present invention was obtained and stored in a cool, dark place in a sealed container for later use. Example 2

[0050] The purpose of this embodiment is to provide a topical composition for improving acne and its preparation method, as detailed below: This topical composition for improving acne is made from the following pharmaceutical ingredients in parts by weight: Group 1: 25 parts of Ampelopsis japonica, 35 parts of Crataegus pinnatifida, 25 parts of Salvia miltiorrhiza, 13 parts of Gardenia jasminoides, and 8 parts of Bletilla striata; Second component: 25 parts mother-of-pearl, 13 parts tribulus terrestris, and 8 parts safflower; The third component consists of 13 parts raw rhubarb, 8 parts gallnut, 8 parts turmeric, 3 parts Ganoderma lucidum, and 3 parts asparagus.

[0051] The origins of the above-mentioned medicinal materials are exactly the same as those described in Example 1, and all comply with the provisions of the Chinese Pharmacopoeia and related standards.

[0052] The preparation method of the topical composition provided in this embodiment is as follows: Accurately weigh each medicinal herb slice according to the above formula, mix them evenly, and put them into the extraction tank.

[0053] First extraction: Add 8 times the amount of purified water to the extraction tank, heat to a gentle boil, extract for 1 hour, filter, collect the first extract, and the separated residue enters the second extraction.

[0054] Second extraction: Add 6 times the amount of purified water to the dregs, heat to a gentle boil and extract for 1 hour, filter, collect the second extract, and the separated residue enters the third extraction.

[0055] Third extraction: Add 6 times the amount of purified water to the dregs, heat to a gentle boil and extract for 1 hour, filter, and collect the third extract.

[0056] The three extracts were combined and filtered through a 200-mesh filter cloth. The filtrate was transferred to a vacuum concentration tank and concentrated under reduced pressure at a temperature of 45-55℃ and a vacuum degree of -0.06 to -0.08 MPa until the relative density of the concentrate reached 1.05 to 1.10 (measured at 45-55℃). After spray drying, the extract powder of the external composition of the present invention was obtained and stored in a cool, dark place in a sealed container for later use.

[0057] Experimental Example 1 Propionibacterium acnes is a Gram-positive anaerobic bacterium. Numerous studies have shown that Propionibacterium acnes is closely related to excessive sebum secretion, hyperkeratosis, inflammation, and tissue damage. This experimental method uses an in vitro method to evaluate the acne-reducing efficacy of the pharmaceutical composition provided by this invention.

[0058] This test example aims to evaluate the antibacterial efficacy of the compositions claimed in this invention using an in vitro method, as follows: 1.1 Experimental Materials and Methods Experimental strain: Propionibacterium acnes; Experimental reagents: phosphate buffer, reinforced Clostridium difficile culture medium (RCM); Equipment: Anaerobic incubator, vertical autoclave; 1.2 Grouping and Sample Preparation: The following groups were set up in this experiment: Sample 1 (Complete Formula): The complete formula of Example 1, namely the first component (30 parts of Ampelopsis japonica, 30 parts of Crataegus pinnatifida, 30 parts of Salvia miltiorrhiza, 15 parts of Gardenia jasminoides, 10 parts of Bletilla striata), the second component (30 parts of pearl powder, 15 parts of Tribulus terrestris, 10 parts of Carthamus tinctorius), and the third component (15 parts of raw rhubarb, 10 parts of Galla chinensis, 10 parts of Curcuma longa, 5 parts of Ganoderma lucidum, 5 parts of Asparagus cochinchinensis), was extracted into powder according to the method of Example 1.

[0059] Sample 2 (first component single use group): only the first component medicinal materials (30 parts of white peony root, 30 parts of hawthorn, 30 parts of salvia miltiorrhiza, 15 parts of gardenia, and 10 parts of bletilla striata) were taken and extracted into powder according to the method of Example 1.

[0060] Sample 3 (second component single use group): only the second component medicinal materials (30 parts of mother-of-pearl, 15 parts of tribulus terrestris, and 10 parts of safflower) were taken and extracted into powder according to the method of Example 1.

[0061] Sample 4 (Third Component Single Use Group): Only the third component medicinal materials (15 parts of raw rhubarb, 10 parts of gallnut, 10 parts of turmeric, 5 parts of Ganoderma lucidum, and 5 parts of asparagus) were taken and extracted into powder according to the method in Example 1.

[0062] Sample 5 (Combined use of first and second components): Take the first and second component medicinal materials (the mass ratio of the two components in the whole formula is the same as that of the first component, i.e., the total amount of the first component is 115 and the total amount of the second component is 55), and extract them into powder according to the method of Example 1.

[0063] Sample 6 (Combined use of first and third components): Take the first and third components of medicinal materials (the mass ratio of the first and third components is the same as the ratio of the two components in the whole formula, that is, the total part of the first component is 115 and the total part of the third component is 45), and extract them into powder according to the method of Example 1.

[0064] Sample 7 (Combined use of second and third components): Take the second and third components of medicinal materials (the mass ratio of the two components in the whole formula is the same as that of the whole formula, that is, the total part of the second component is 55 and the total part of the third component is 45), and extract them into powder according to the method of Example 1.

[0065] All the above sample extract powders were prepared according to the method described in Example 1 (water decoction extraction, concentration, and drying). All samples were dissolved and diluted with phosphate-buffered saline (PBS) to the same dosage concentration before testing. The dosage concentration was calculated based on the raw medicinal material and was equivalent to a raw medicinal material concentration of 150 mg / mL.

[0066] 1.3 Experimental Methods: Propionibacterium acnes in the logarithmic growth phase was diluted with PBS to prepare approximately 10 6 CFU / mL bacterial suspension. The extract powders of the above samples were dissolved in PBS and diluted to the same concentration (crude drug concentration of 150 mg / mL) to obtain sample solutions.

[0067] Mix 0.5 mL of bacterial suspension and 0.5 mL of sample solution in a sterile centrifuge tube and incubate at room temperature for 15 minutes. Use PBS as a blank control (control group) instead of the sample. After incubation, perform a 10-fold serial dilution of 100 μL of the mixture, and plate the appropriate dilution onto RCM plates, with three replicates for each dilution. Incubate the plates in an anaerobic incubator at 37°C for 48 hours, and then count the viable bacteria. Repeat the experiment three times. The inhibition rate is calculated using the following formula: Antibacterial rate (%) = (average colony count of control sample group - average colony count of test sample group) / average colony count of control sample group × 100%.

[0068] The antibacterial rate of the test sample is >50%, which indicates that the sample has a certain acne-removing effect. The higher the antibacterial rate of Propionibacterium acnes, the stronger the acne-removing effect of the sample.

[0069] 1.4 Experimental Results and Analysis The experimental results are shown in Table 1.

[0070] Table 1. Inhibition rate of each sample against Propionibacterium acnes (n=3) As shown in Table 1, under the same total drug concentration, the total formulation of this invention (Sample 1) exhibited a high inhibition rate of 99.89% against Propionibacterium acnes, with a surviving bacterial count of only 40 CFU / mL, demonstrating extremely strong in vitro antibacterial activity. Among the single-component formulations, the third component (Sample 4) showed the strongest antibacterial activity, with an inhibition rate of 98.97%; the first component (Sample 2) and the second component (Sample 3) also exhibited good antibacterial effects, with inhibition rates of 85.29% and 79.71%, respectively.

[0071] It is noteworthy that the antibacterial rate of the combination of the first and second components (Sample 5) (71.43%) was lower than that of either component used alone, suggesting that at this total concentration, the combined use of the first and second components may lead to a decrease in antibacterial effect due to interactions between the components. However, when the third component was added to the formulation, the situation changed significantly: the antibacterial rate of the combination of the first and third components (Sample 6) rebounded to 99.49%, and the antibacterial rate of the combination of the second and third components (Sample 7) reached 99.20%, both approaching the level of the complete formulation. The complete formulation (Sample 1) achieved the best antibacterial effect (99.89%), with the lowest number of surviving bacteria.

[0072] Experimental Example 2 This experiment used a transgenic neutrophil-based green fluorescent zebrafish model to evaluate the inhibitory effect of each sample on the inflammatory response induced by Propionibacterium acnes in vivo. The preparation of all samples was the same as in Experiment 1.

[0073] 2.1 Experimental Materials and Methods Experimental system: transgenic neutrophil-containing green fluorescent zebrafish (MPX).

[0074] Zebrafish age: 3 days post-fertilization (3 dpf).

[0075] Sample size per group: 15 tails.

[0076] Adult fish rearing and breeding methods: The rearing and breeding methods shall be carried out in accordance with laboratory standards and meet the requirements of international AAALAC certification (certification number: 001458).

[0077] 2.2 Grouping and Sample Preparation The following samples were prepared according to the formulation and extraction process of Example 1 and then freeze-dried into powder: Sample 1 (Full Formula): Extracted by mixing and extracting the first, second and third components according to the optimal prescription in Example 1.

[0078] Sample 2 (Single use of the first component): Extraction was performed using only the first component of the drug (30g of Ampelopsis japonica, 30g of Crataegus pinnatifida, 30g of Salvia miltiorrhiza, 15g of Gardenia jasminoides, and 10g of Bletilla striata).

[0079] Sample 3 (second component single use group): only the second component drugs (30g of mother-of-pearl, 15g of tribulus terrestris, and 10g of safflower) were extracted.

[0080] Sample 4 (Third Component Single Use Group): Only the third component drugs (raw rhubarb 15g, gallnut 10g, turmeric 10g, Ganoderma lucidum 5g, asparagus 5g) were extracted.

[0081] Sample 5 (Combined use of first and second components): The first and second components of the drug were mixed and extracted.

[0082] Sample 6 (Combined use of first and third components): The first and third components of the drug were mixed and extracted.

[0083] Sample 7 (Combined use of second and third components): The second and third components of the drug were mixed and extracted.

[0084] 2.3 Experimental Methods Healthy zebrafish were randomly selected, and except for the normal control group, the other groups were administered 10 mg / L via microinjection. 9 A local inflammation model was established in zebrafish by injecting 10 nL of Propionibacterium acnes bacterial suspension at CFU / mL, while the normal control group was injected with an equal volume of PBS. After modeling, each sample group was given the corresponding sample test solution, and the dosage was kept consistent across groups based on the amount of raw drug.

[0085] 2.3.1 Dosage exploration To determine the appropriate drug concentration for this experimental system, a series of raw drug concentration gradients were first set up for sample 1 (the whole formula group) for preliminary experiments. According to the raw drug amount, the drug concentration range covered 0.083 mg / mL to 6.6 mg / mL, and 0.005% salicylic acid solution was used as a positive control.

[0086] 2.3.2 Acne-removing efficacy experiment Specific drug administration procedure: Add 100 μL of the corresponding concentration of sample test solution to each well, and then make up to a final volume of 3 mL with physiological saline; add an equal volume of physiological saline to the model control group and the normal control group. Incubate at 28°C in the dark for 3 hours.

[0087] After incubation, 10 zebrafish were randomly selected from each group and photographed under a fluorescence microscope. Some typical images of acne removal are shown below. Figure 1 As shown, in this Figure 1 In the image, the area within the yellow dashed box represents the neutrophil counting region. The number of neutrophils (N) in the zebrafish yolk sac was analyzed using NIS-Elements image analysis software. The formula for calculating the acne-reducing efficacy is as follows: Acne-removing efficacy (%) = [N(model control group) - N(sample group)] / [N(model control group) - N(normal control group)] × 100%.

[0088] Statistical analysis was performed using GraphPad Prism software with t-tests, and p < 0.05 was considered statistically significant.

[0089] 2.4 Experimental Results and Analysis 2.4.1 Study on dose-response relationship of the full-formula group First, the inhibitory effect of sample 1 on the inflammatory response induced by Propionibacterium acnes was investigated at multiple safe drug concentrations. The experimental results are shown in Table 2.

[0090] Table 2. Effects of different concentrations of crude drug on the number of neutrophils in zebrafish and its acne-reducing efficacy in Sample 1 (n=10) As shown in Table 2, Sample 1 significantly inhibited neutrophil aggregation induced by Propionibacterium acnes, exhibiting a favorable dose-response relationship. The normal control group showed very few neutrophils, while the model control group showed a significantly increased number, indicating successful propionibacterium acnes modeling. The positive control group (salicylic acid) significantly reduced the number of neutrophils.

[0091] At a crude drug concentration of 5.0 mg / mL, Sample 1 showed an acne-reducing efficacy of 84%, with neutrophil counts decreasing to near-normal levels, demonstrating highly effective anti-inflammatory activity. When the concentration increased to 6.6 mg / mL, the efficacy was 62%, slightly lower than the 5.0 mg / mL group, but still significantly better than the model control group. When the concentration decreased to 3.3 mg / mL, the efficacy was 59%, maintaining a high inhibition rate. As the concentration further decreased, the efficacies of the 1.7 mg / mL, 0.83 mg / mL, and 0.33 mg / mL groups were 47%, 43%, and 41%, respectively, with the inhibitory effect gradually weakening; while at 0.17 mg / mL and below, the efficacy decreased to 39% and 23%, respectively, with neutrophil counts approaching the level of the model control group, and the inhibitory effect essentially disappeared.

[0092] The above results indicate that sample 1 has a concentration-dependent inhibitory effect on the inflammatory response induced by Propionibacterium acnes, with 5.0 mg / mL being the optimal effective concentration. Considering both efficacy and safety, a crude drug concentration of 5.0 mg / mL was selected as the uniform test concentration for all samples in subsequent studies.

[0093] 2.4.2 Comparison of anti-inflammatory activities among different groups in the fractional formulation study To further observe the compatibility effects of each medicinal ingredient in this invention, we conducted a decomposition study on the optimal prescription described in Example 1. Based on the concentration exploration and dose-effect relationship study results in section 2.4.1 above, Sample 1 (whole extract) exhibited the best safety and anti-inflammatory activity at a crude drug concentration of 5.0 mg / mL. Therefore, in this part of the experiment, 5.0 mg / mL was selected as the uniform test concentration for all samples to compare the in vivo anti-inflammatory activity of different combinations under the same conditions. The experimental results are shown in Table 3, and the following groups were tested under the same conditions.

[0094] Table 3. Acne-reducing efficacy of each sample group in the zebrafish model (n=10) As shown in Table 3, at the same total drug concentration (5.0 mg / mL), compared with the model control group, the number of neutrophils in the zebrafish yolk sac of all sample groups was significantly reduced, indicating that each sample had in vivo anti-inflammatory activity and could effectively inhibit the inflammatory response induced by Propionibacterium acnes. Among them, sample one of the present invention had the highest acne-removing efficacy, reaching 85% (slightly different from 84% in section 2.4.1, due to rounding in data calculation and normal experimental fluctuations), which was not only much higher than the first, second, and third components used alone, but also significantly higher than the group using any two components in combination. This result clearly shows that, under the complex physiological environment in vivo, the anti-inflammatory effect obtained by the combined use of the first, second, and third components is far superior to any single component or combination of two components.

[0095] It is particularly noteworthy that while the combined efficacy of the first and second components was higher than that of either component alone, it was lower than that of the complete formula. However, the efficacy of the complete formula jumped significantly to 85% after the introduction of the third component. This suggests that the third component not only has its own anti-inflammatory activity, but more importantly, it can produce a synergistic effect with the first and second components, thereby maximizing the overall anti-inflammatory capacity. Example 3

[0096] The purpose of this embodiment is to provide an acne-reducing gel formulation comprising the topical composition of the present invention, and to observe its effect on improving acne through human trials.

[0097] 3.1 Preparation of Acne-Removing Gel According to the optimal prescription described in Example 1 (first component: 30 g of Ampelopsis japonica, 30 g of Crataegus pinnatifida, 30 g of Salvia miltiorrhiza, 15 g of Gardenia jasminoides, 10 g of Bletilla striata; second component: 30 g of pearl powder, 15 g of Tribulus terrestris, 10 g of Carthamus tinctorius; third component: 15 g of Rheum palmatum, 10 g of Galla chinensis, 10 g of Curcuma longa, 5 g of Ganoderma lucidum, 5 g of Asparagus cochinchinensis), the raw medicinal materials were weighed, with a total amount of 215 g. The extract powder of the external composition of this invention was obtained according to the extraction method described in Example 1 (three decoctions, combined filtrates, concentrated under reduced pressure to a relative density of 1.05-1.10 (measured at 45-55℃), and then spray-dried). An appropriate amount of the extract powder was dissolved in water and brought to a final volume so that each 30 g solution contained the extract equivalent to 50 g of raw medicinal materials, thus obtaining the standard stock solution for gel preparation.

[0098] The formula for the acne gel, by weight parts, is as follows: Phase A (gel matrix): Carbomer 940 0.5 parts, glycerol 5.0 parts, purified water to make up to 100 parts.

[0099] Phase B (Efficacy components and stabilizers): 20.0 parts of the standard mother liquor of the Chinese herbal extract prepared according to the above method (each 30g corresponds to 50g of raw medicinal material), and 0.03 parts of disodium ethylenediaminetetraacetate.

[0100] Phase C (pH adjuster): Arginine, appropriate amount; Phase D (preservative): 0.2 parts of SymDiol® 68 (a mixture of 1,2-hexanediol, octyl glycol, and ethylhexylglycerin).

[0101] The preparation steps are as follows: (1) Slowly sprinkle Carbomer 940 from phase A into purified water, add glycerol, stir evenly, and let it stand to swell overnight to obtain a transparent gel matrix.

[0102] (2) Add disodium ethylenediaminetetraacetate from phase B to the standard mother liquor of traditional Chinese medicine extract and stir until completely dissolved.

[0103] (3) Add the mixture obtained in (2) slowly to the gel matrix in (1) while stirring, and stir until homogeneous.

[0104] (4) Add C-phase arginine and adjust the pH of the system to 5.5-6.5 to form a uniform and transparent gel.

[0105] (5) Finally, add the D-phase preservative and stir evenly to obtain the acne-removing gel of the present invention (full formula sample).

[0106] 3.2 Preparation of control samples In addition, this embodiment also provides several control samples. The gel matrix formulation (phases A, C, and D) and preparation steps of the control samples are exactly the same as those in 3.1, except that the source of the standard mother liquor of the Chinese herbal extract in phase B is different. The mother liquor used for each control sample was prepared independently according to the principle of "50 g of total crude drug, extracted, concentrated, and diluted with water to 30 g". The specific composition and formulation are as follows: Five sample groups (combined first and second components): Component 1 (Ampelopsis japonica, Crataegus pinnatifida, Salvia miltiorrhiza, Gardenia jasminoides, Bletilla striata, mass ratio 30:30:30:15:10) and Component 2 (Mother-of-pearl, Tribulus terrestris, Carthamus tinctorius, mass ratio 30:15:10), with the same mass ratio as the first component:second component in the whole formula = 115:55. Raw medicinal materials were weighed according to this ratio to make a total raw medicinal material weight of 50 g. Extraction and concentration were performed according to the method in Example 1, and water was added to a final volume of 30 g to obtain the first + second component mother liquor.

[0107] The sample consisted of six groups (a combined group of the first and third components): the first component (Ampelopsis japonica, Crataegus pinnatifida, Salvia miltiorrhiza, Gardenia jasminoides, Bletilla striata, in a mass ratio of 30:30:30:15:10) and the third component (Rheum palmatum, Galla chinensis, Curcuma longa, Ganoderma lucidum, Asparagus cochinchinensis, in a mass ratio of 15:10:10:5:5), with the same mass ratio as the first component:third component in the whole formula (115:45). The raw medicinal materials were weighed according to this ratio to make a total of 50 g of raw medicinal materials. Extraction and concentration were performed according to the method in Example 1, and water was added to bring the volume to 30 g to obtain the mother liquor of the first and third components.

[0108] Seven sample groups (combined group of second and third components): the second component (mother-of-pearl, tribulus terrestris, safflower, mass ratio 30:15:10) and the third component (raw rhubarb, gallnut, turmeric, ganoderma lucidum, asparagus, mass ratio 15:10:10:5:5), with the same mass ratio of the second component to the third component in the whole formula as 55:45. Raw medicinal materials were weighed according to this ratio to make a total raw medicinal material weight of 50 g. Extraction and concentration were performed according to the method in Example 1, and water was added to a final volume of 30 g to obtain the mother liquor of the second and third components.

[0109] 3.2.1 Selection and grouping of subjects: This study recruited 80 participants aged 18-35 years with mild to moderate acne on the face or back (meeting Pillsbury classification I-III), regardless of gender. Exclusion criteria included: systemic or topical use of antibiotics, retinoids, and corticosteroids within the past month; pregnant and lactating women; and known allergies to cosmetic ingredients. Participants were randomly assigned to four groups of 20 each using a random number table, corresponding to the following treatments: Sample 1 (full treatment group), Sample 5 (combined group of components 1 and 2), Sample 6 (combined group of components 1 and 3), and Sample 7 (combined group of components 2 and 3).

[0110] During the study, a total of 8 participants dropped out for personal reasons (not completing the entire trial), resulting in 72 valid cases. The completion rates for each group were as follows: Group 1: 19 participants, Group 5: 17 participants, Group 6: 18 participants, and Group 7: 18 participants. The experimental results are based on the statistical results of those who completed the study.

[0111] 3.2.2 Usage Instructions: After cleansing their face / back every morning and evening, participants applied an appropriate amount of the corresponding acne-reducing gel evenly to the acne lesions and gently massaged until absorbed. They used the gel continuously for 10 days, during which time they were instructed to maintain a normal sleep schedule and avoid spicy and irritating foods.

[0112] 3.2.3 Evaluation of efficacy and safety Follow-up photos were taken before use (day 0) and on day 10 after use to record changes in skin lesions and adverse reactions. A dermatologist evaluated the skin lesions based on a score. The weighting of each skin lesion is as follows: Acne (whiteheads, blackheads): 0.5 points each; Inflammatory papules: 1 point each; Pustules: 2 points each; Nodules / cysts: 3 points each; The efficacy index was calculated based on the changes in the subjects' scores before and after treatment, using the following formula: Therapeutic efficacy index (%) = (Total score before treatment - Total score after treatment) / Total score before treatment × 100%. Based on the therapeutic efficacy index, the degree of skin lesion improvement is divided into three levels: Significant improvement: Therapeutic effect index ≥50%; Slight improvement: 20% ≤ efficacy index < 50%; Essentially ineffective: efficacy index <20%.

[0113] 3.2.4 Experimental Results (1) Safety evaluation results During the 10-day study, all 72 subjects who completed the full course of treatment showed good skin tolerance.

[0114] ① Clinical observation record: Based on clinical examination by dermatologists and subjective feedback from subjects, no objective irritation symptoms such as redness, swelling, erythema, or papules appeared on any of the test sites, nor were there any subjective discomforts such as itching, burning, stinging, or tightness.

[0115] ②Incidence of adverse reactions: During the trial, the incidence of skin irritation and allergic adverse reactions was 0%.

[0116] The results of the study indicate that the acne-reducing gel composition described in this application is mild and non-irritating, with excellent skin clinical safety and compatibility, and is suitable for long-term or continuous application to the face.

[0117] (2) Results of efficacy evaluation The skin lesion scores and improvement status before and after treatment in each group are shown in Tables 4 and 5.

[0118] Table 4 Comparison of skin lesion scores before and after treatment in each group (x±s) Table 4 shows that the skin lesion scores of all groups decreased significantly after treatment compared to before treatment (paired t-test, p<0.05), indicating that the preparations in each group showed an effect of improving acne within a short period of 10 days. In terms of efficacy index, Sample 1 (the complete formula group) had the highest rate at 71.2%; Sample 6 was second at 43.9%; Sample 5 and Sample 7 were 30.2% and 27.1%, respectively. The data suggest that the complete formula group showed the most significant improvement.

[0119] Table 5. Distribution of Improvement Levels in Each Group As shown in Table 5, the significant improvement rate in Sample 1 (the full-strength group) was as high as 89.5% (17 / 19), the slight improvement rate was 10.5% (2 / 19), and the essentially ineffective rate was 0; that is, almost all subjects achieved significant improvement. In contrast, the significant improvement rates in Sample 5 and Sample 7 were only 17.6% and 11.1%, respectively, with essentially ineffective rates as high as 29.4% and 38.9%; the improvement in Sample 6 was in the middle, with a significant improvement rate of 38.9%.

[0120] The above experimental results show that the complete formula provided by the present invention has the best effect on improving acne in the human body, and it takes effect quickly. Significant therapeutic effects can be observed within 10 days, which shows the scientific nature and superiority of the formulation of the topical composition of the present invention.

[0121] Some typical cases can be found Figures 2-3 ,in: Typical Case 1: Ms. Wang, female, 26 years old, office worker (sample group). Before use, multiple obvious inflammatory red papules were visible between her eyebrows and around her lips, accompanied by mild tenderness, and her skin was oily (see...). Figure 2 (Left). The acne-removing gel of this invention was used twice daily. After 8 days of continuous use, the red papules on the face had largely subsided, and no obvious pigmentation was observed. The oiliness of the skin in the test area improved, and the skin texture became more stable (see...). Figure 2 (Right). No adverse reactions were observed during treatment.

[0122] Typical Case 2: Mr. Zhao, male, 24 years old, office worker (sample group). He had recurrent acne on his jawline for over a year, which had recently worsened. Upon examination, his jawline showed densely distributed inflammatory papules, significant cystic infiltration, and redness, accompanied by significant itching and tenderness (see...). Figure 3 (Left). The acne-removing gel of this invention was used twice daily. A follow-up examination on the 5th day of continuous use showed a significant reduction in redness and swelling of the jawline, and the cysts began to soften and shrink. While eliminating active skin lesions, it also significantly improved early acne scars and post-inflammatory hyperpigmentation, with no new hyperpigmentation observed. Furthermore, the subjects reported that itching and pain had essentially disappeared (see...). Figure 3 right).

[0123] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the present invention, and all such modifications and substitutions should be covered within the scope of the claims of the present invention. Technical aspects, shapes, and structures not described in detail in this invention are all well-known technologies.

Claims

1. An external use composition for improving acne, characterized by comprising, The topical composition is made from the following pharmaceutical ingredients: Group 1: 25-35 parts of Ampelopsis japonica, 25-35 parts of Crataegus pinnatifida, 25-35 parts of Salvia miltiorrhiza, 13-17 parts of Gardenia jasminoides, and 8-12 parts of Bletilla striata; Second component: 25-35 parts mother-of-pearl, 13-17 parts tribulus terrestris, 8-12 parts safflower; The third component consists of 13-17 parts of raw rhubarb, 8-12 parts of gallnut, 8-12 parts of turmeric, 3-7 parts of Ganoderma lucidum, and 3-7 parts of asparagus.

2. An external use composition for improving acne, characterized by, The topical composition is made from the following pharmaceutical ingredients: 25-35 parts of Ampelopsis japonica, 25-35 parts of Crataegus pinnatifida, 25-35 parts of Salvia miltiorrhiza, 13-17 parts of Gardenia jasminoides, and 8-12 parts of Bletilla striata.

3. An external use composition for improving acne, characterized by, The topical composition is made from the following pharmaceutical ingredients: 25-35 parts of mother-of-pearl, 13-17 parts of tribulus terrestris, and 8-12 parts of safflower.

4. A topical composition for improving acne, characterized in that, The topical composition is made from the following pharmaceutical raw materials: 13-17 parts of raw rhubarb, 8-12 parts of gallnut, 8-12 parts of turmeric, 3-7 parts of Ganoderma lucidum, and 3-7 parts of asparagus.

5. A method for preparing the topical composition according to any one of claims 1-4, characterized in that, The method includes: mixing the pharmaceutical raw materials, extracting with water, concentrating to obtain an extract paste of the topical composition, or further spray-drying or freeze-drying the extract paste to obtain an extract powder.

6. The use of the topical composition according to any one of claims 1-5 in the preparation of cosmetics or pharmaceuticals for the prevention, relief and / or treatment of acne.

7. The application according to claim 6, characterized in that, The acne includes one or more of comedones, inflammatory papules, pustules, nodules, and cysts; and / or the application also includes improving post-acne erythema, post-acne hyperpigmentation, and post-acne scars.

8. The application according to claim 6, characterized in that, The topical composition has the effect of inhibiting the activity of Propionibacterium acnes and / or inhibiting the inflammatory response induced by Propionibacterium acnes.

9. A cosmetic product for improving acne, characterized in that, The cosmetic is prepared as a serum, gel, lotion, cream, mask, lotion, spray, or jelly, comprising an extract extract or dried extract powder of any one of the topical compositions as described in claims 1-4 as an active ingredient.

10. A medicine for improving acne, characterized in that, The active ingredient comprises an extract extract or dried extract powder thereof as described in any one of claims 1-4, and pharmaceutically acceptable excipients.

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