Acid-free acne-removing composition and its application, daily chemical product

Abstract

The present application belongs to the technical field of cosmetic production, and discloses an acid-free acne-removing composition, comprising 10-20% of dipeptide-15, 1-5% of olive leaf extract, 5-10% of acetyl glucosamine, 0.1-1% of 10-hydroxydecanoic acid, 5-10% of yeast / zinc fermentation product, 1-5% of bisabolol, 5-10% of tetrahydromethylpyrimidine carboxylic acid, 1-10% of capryloyl glycine, 0.01-1% of polylysine, 28-32% of polyol and the balance of deionized water. Through the synergistic effect among the above multiple components, the antibacterial ability of the composition and the inhibitory effect on inflammatory factors 1L-1α and 5α-reductase are further improved. Moreover, the composition disclosed in the present application also has good anti-allergic ability. In addition, the present application also discloses the application of the acid-free acne-removing composition and a daily chemical product.

Description

Technical Field

[0001] The invention relates to the technical field of cosmetic production, and in particular to an acid-free acne-removing composition and application thereof, and daily chemical products. Background Art

[0002] The Guidelines for Primary Diagnosis and Treatment of Acne Vulgaris (2023) and the Guidelines for the Treatment of Acne in China (2019 Revised Edition) both list the definition, pathogenesis, classification and corresponding treatment methods of acne. Acne vulgaris, also known as "acne", is a chronic inflammatory skin disease involving the hair follicles and sebaceous glands. It is prone to occur on the face, chest, and back. Clinically, it mainly manifests as acne, papules, pustules, cysts or nodules, often accompanied by enlarged pores and sebum secretion. Acne vulgaris is common in adolescent boys and girls, with an incidence rate of up to 93% in adolescents. It has become the eighth largest chronic disease in the world, and is prone to post-inflammatory erythema and pigmentation. According to statistics, 31.8% of acne vulgaris can cause secondary sensitive skin, and 3% to 7% can leave scars, which seriously affect the patient's appearance and physical and mental health.

[0003] The pathogenesis of acne is still not fully elucidated. Under the genetic background, hormone-induced excessive secretion of lipids from sebaceous glands, abnormal keratinization of the sebaceous gland ducts, proliferation of hair follicle microorganisms such as Propionibacterium acnes, and inflammation and immune responses are related to it. Genetic factors play an important role in the occurrence of acne, especially severe acne; androgen is the main inducing factor for sebaceous gland hyperplasia and massive secretion of lipids. Other hormones such as insulin-like growth factor-1 (IGF-1) and insulin growth hormone may also be related to the occurrence of acne; large secretion of lipids by sebaceous glands is considered to be a prerequisite for the occurrence of acne, but changes in lipid composition such as increased content of squalene peroxide, wax esters, free fatty acids, increased proportion of unsaturated fatty acids, and decreased linoleic acid are also important factors leading to the occurrence of acne; hair follicle microorganisms such as Propionibacterium acnes participate in the occurrence and development of acne through innate immunity and acquired immunity. Abnormal keratinization of the sebaceous gland ducts, inflammation and immune response are the main pathological characteristics of acne, and the inflammatory response runs through the entire process of the disease. Hair follicle microorganisms and abnormal lipids activate Toll-like receptors (TLRs) to produce interleukin (IL)-1α and other related inflammatory mediators. IL-1α is currently considered to be the main factor in sebaceous gland duct keratinization and the formation of microacne and acne. As the disease progresses, a large amount of lipids accumulate, leading to further proliferation of lipophilic and anaerobic Propionibacterium acnes, and acquired immunity is activated. The increasing inflammatory response induces rupture of the hair follicle wall, and lipids, microorganisms, and hair enter the dermis, producing a foreign body-like reaction. After the acne lesions subside, erythema pigmentation and scar formation are often left behind, which is closely related to the severity of acne, individual differences or improper treatment.

[0004] For the treatment of acne in special populations (children, pregnant or lactating), the U.S. Food and Drug Administration (FDA) has approved a combination of 2.5% benzoyl peroxide & 1% adapalene gel for children aged ≥9 years, and 0.05% tretinoin gel for children aged ≥10 years. It is safe to use topical azelaic acid and clindamycin in pregnant or lactating populations (pregnancy classification B). It is also recommended to use some clinically proven anti-acne functional skin care products as an adjuvant. It is worth noting that the use of topical treatments with tretinoin (including adapalene gel), antibacterial drugs such as benzoyl peroxide, and antibiotic drugs such as clindamycin is often accompanied by varying degrees of skin irritation reactions, and long-term use during the treatment period can easily lead to skin barrier damage. At the same time, topical drugs alone usually only act on 1 to 2 of the four main pathogenic links of acne, and only combined use can increase its action links to 2 to 3. Based on the psychological and physiological safety needs of special populations, a non-drug, mild and non-irritating cosmetic product that simultaneously covers four action pathway targets is of great development significance.

[0005] Chinese patent application 201910528459.1 discloses a blue light protection cosmetic composition and a preparation method thereof and a cosmetic containing the composition. The composition provided by the scheme includes sunscreen, whitening agent, anti-aging agent and other ingredients. Further reading of the specification of the scheme shows that the whitening agent component in the scheme can be selected from a variety of plant extracts or organic acids, etc. At the same time, the scheme further optimizes the composition and also includes a moisturizer component, and also explains that the moisturizer can be selected from acetyl glucosamine;

[0006] It can be seen that the above scheme discloses some components in the present application. On the one hand, although the whitening agent contains a variety of plant extracts, it does not contain the olive leaf extract used in the present application. On the other hand, the plant extract appears more as a whitening agent in the scheme, while acetyl glucosamine appears as a moisturizer. From the embodiments and comparative examples of the scheme, we cannot infer that there is some synergistic effect between acetyl glucosamine and the plant extract.

[0007] Chinese patent application 202110648670.4 discloses an anti-aging moisturizing composition and its application. The anti-aging moisturizing composition includes: aminobutyric acid, ectoine, saccharide isomers and bisabolol; but it can be seen that the scheme only discloses some components of the present application, and from the experimental results of the scheme, it can be seen that ectoine and bisabolol need to be used together with aminobutyric acid and saccharide isomers in the scheme to achieve good anti-allergic experimental effects and inhibition of proteases.

[0008] Chinese patent application 201910752753.0 discloses a safe and gentle acne-removing essence and a preparation method. The essence contains water, polyacrylate cross-linked polymer-6, disodium EDTA, quaternary ammonium salt-73, cucumber fruit extract, 1,2-hexanediol, glycerin, butylene glycol, spiraea ulmaria extract, lentil seed extract, p-hydroxyacetophenone, hydrolyzed pansy extract, phenoxyethanol, ethylhexylglycerin, multiple amino acids, tremella extract, sodium hyaluronate and betaine. At the same time, it can be seen from the embodiments of the scheme that the scheme has a good acne-removing effect, but the scheme still only discloses part of the components of the scheme, and the components used in the scheme are of various types.

[0009] The problem that this scheme needs to solve is: how to develop an acid-free acne-removing composition to inhibit the secretion of the inflammatory factor IL-1α, thereby achieving the effect of acne removal. Summary of the invention

[0010] The purpose of the present invention is to provide an acid-free acne-removing composition, which does not contain inorganic acid, thereby reducing the irritation of inorganic acid to the skin. At the same time, the acid-free acne-removing composition can inhibit the secretion of inflammatory factor IL-1α, thereby achieving the effect of acne removal.

[0011] To achieve the above object, the present application discloses an acid-free acne-removing composition, which specifically comprises the following components by mass fraction:

[0012]

[0013]

[0014] Preferably, the polyol is selected from at least one of glycerol, butylene glycol, dipropylene glycol, 1,3-propanediol and methylpropylene glycol.

[0015] In addition, the present application also discloses the use of the acid-free acne-removing composition as described above as an acne-removing active ingredient in daily chemical products.

[0016] In addition, the present application also discloses a daily chemical product containing 8-12%wt of the above-mentioned acid-free acne-removing composition.

[0017] Preferably, the daily chemical product is lotion, essence or facial mask.

[0018] Preferably, the dosage form of the daily chemical product is a spray, lotion or cream.

[0019] The beneficial effects of this application are:

[0020] The dipeptide-15 contained in the acid-free acne-removing composition of the present application can regulate the intracellular chloride ion concentration and reduce the influx of calcium ions by inhibiting unsaturated fatty acid signals, thereby restoring the difference in calcium ion concentration inside and outside the keratinocytes to a normal level, thereby reducing the occurrence of abnormal keratinocyte membrane potential, imbalanced differentiation and proliferation, and the like, thereby achieving the effect of shrinking pores;

[0021] Olive leaf extract contains oleuropein, which has antifungal, anti-inflammatory, antiviral, antioxidant, anti-cancer and hypoglycemic effects. Oleuropein also has strong antioxidant activity and anti-inflammatory (1L-1α) in vitro, and inhibits Propionibacterium acnes. Its strong antioxidant activity comes from its dihydroxy-substituted structure, which can bind superoxide anions, hydrogen peroxide, nitric oxide free radicals and nitro free radicals to scavenge free radicals. At the same time, hydroxytyrosol, another major active ingredient in olive leaf extract, has a catechol structure and is a typical antioxidant stress substance and free radical scavenger.

[0022] Acetyl glucosamine is a monosaccharide derived from glucose. It is a precursor of hyaluronic acid and the basic component of many important polysaccharides in biological cells. It has excellent transdermal absorption and can promote skin hydration. Acetyl glucosamine can stimulate the differentiation of basal cells and change the way cells differentiate and adhere, thereby inhibiting the re-aggregation of keratinocytes that have been separated, accelerating the opening process of the skin "hinge" and promoting skin shedding, so as to achieve a mild exfoliating effect.

[0023] 10-Hydroxydecanoic acid has the advantages of antibacterial, anti-inflammatory, anti-tumor and immune regulation. In the oil control pathway, it has a significant effect on the inhibition of 5α-reductase; 5α-reductase can catalyze the conversion of testosterone into dihydrotestosterone, which binds to the androgen receptors of sebaceous gland cells to promote the development of sebaceous glands and produce a large amount of sebum. The inhibition rate of 5α-reductase reflects its ability to inhibit excessive sebum secretion;

[0024] On the one hand, yeast / zinc fermentation products have many beneficial effects on the skin, such as anti-inflammatory, acne removal, oil control, and reduction of dandruff. On the other hand, zinc specifically participates in and promotes the cytokine messenger that communicates between different cells (such as mast cells, platelets, macrophages, neutrophils, natural killer cells, and lymphocytes) during the inflammatory process. In addition, zinc ions can block the influx of calcium ions into cells through glutamate receptors, thereby preventing cells from abnormal differentiation and proliferation.

[0025] Bisabolol relieves inflammatory responses by inhibiting the release of 5-lipoxygenase and IL-1a, IL-6 and TNF-α factors, and has multiple effects such as soothing, anti-oxidation, anti-infection, increasing bioavailability, and whitening.

[0026] Ectohydromethylpyrimidine carboxylic acid significantly increased the expression of FLG protein (a protein related to skin barrier repair) in the SLS injury model, and had a certain effect on repairing skin barrier damage; Ectohydromethylpyrimidine carboxylic acid could also significantly reduce the release of IL-1β verification factor secreted by RAW264.7 cells induced by LPS;

[0027] Capryloyl glycine has good skin affinity and can significantly inhibit the activity of 5α-reductase. It can also inhibit the proliferation of anaerobic microorganisms related to unbalanced skin caused by acne. Capryloyl glycine can also create a slightly acidic environment to help acne-prone skin restore a healthy skin acid barrier.

[0028] At the same time, compared with the few "acid-free" acne-removing products on the market, the composition disclosed in the present application further avoids organic acid components such as lactic acid that may be contained in fermentation ingredients (such as lactobacillus fermentation extract and yeast fermentation extract), as well as residual azelaic acid components in the raw materials of azelaic acid derivatives (zelaic acid amide MEA, potassium azeloyl glycinate), and possible salicylic acid components in white willow bark fermentation extract. The present application provides an acid-free acne-removing composition with four complete action pathways, which is more in line with the acne removal needs of pregnant women, teenagers and people with highly sensitive skin. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a comparison diagram of the sample of Example 3 used on the right face of subject A, before and after D0 and D28;

[0030] Figure 2 This is the before and after comparison of D0 and D28 for the left face of subject A without using the sample;

[0031] Figure 3 This is a comparison diagram of the sample of Example 3 used on the right face of subject B, before and after D0 and D28;

[0032] Figure 4 This is the before and after comparison between D0 and D28 for the left face of subject B without using the sample. DETAILED DESCRIPTION

[0033] The present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. In the description of the present invention, it should be noted that, if no specific conditions are specified in the embodiments, the conditions are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0034] Before describing the examples, the following necessary descriptions are first given for the raw materials used in the examples:

[0035] Dipeptide-15: also known as N-glycylglycine, diglycine, diglycine, molecular formula C4H8N2O3, molecular weight 132Da, purchased from Shenzhen Weiqi Company;

[0036] Olive leaf extract: extracted from the leaves of olive trees, purchased from Hallstar Company, USA;

[0037] Acetyl glucosamine: also known as N-acetylglucosamine (NAG), the molecular formula of acetyl glucosamine is C8H 15 NO6, molecular weight 221Da, purchased from Shandong Huaxi Biotechnology Co., Ltd.;

[0038] 10-Hydroxydecanoic acid: also known as royal acid, molecular formula is C 10 H 20 O3, molecular weight 188Da, purchased from Guangzhou Miaosen Company;

[0039] Yeast / zinc fermentation product: obtained by culturing yeast using zinc-containing culture medium, for details, please refer to patent CN101575580A, purchased from Hubei Angel Yeast Company;

[0040] Bisabolol: purchased from Givaudan, Switzerland;

[0041] Tetrahydromethylpyrimidine carboxylic acid: molecular formula is C6H 10 N2O2, molecular weight 142Da, purchased from Shandong Yimingtai Company;

[0042] Capryloylglycine: purchased from SEPPIC, France

[0043] Polylysine: The structural unit is lysine, the degree of polymerization is 48, and it was purchased from IFF Company in the United States.

[0044] Preparation process of acid-free acne-removing composition:

[0045] Step 1: Weigh capryloyl glycine, heat it to 85°C with butanediol, stir and dissolve it completely, and cool it to room temperature to obtain the first phase;

[0046] Step 2: Weigh dipeptide-15, acetyl glucosamine, and tetrahydroxymethylpyrimidine carboxylic acid, heat to 45°C with deionized water, and stir to dissolve completely to obtain the second phase;

[0047] Step 3: Mix the first phase and the second phase, then add olive leaf extract, bisabolol, 10-hydroxydecanoic acid, yeast / zinc fermentation product, and polylysine one by one, and mix them evenly at a low speed to obtain an acid-free acne-removing composition.

[0048] Examples 1-3

[0049] An acid-free acne-removing composition, the formula of which is shown in Table 1:

[0050] Table 1

[0051]

[0052]

[0053] Comparative Examples 1-13

[0054] A composition, the formula is shown in Table 2:

[0055] Table 2

[0056]

[0057]

[0058] Application examples:

[0059] The compositions obtained in the examples and comparative examples are prepared into finished essences:

[0060] The formula of the essence is shown in Table 3:

[0061] Table 3

[0062]

[0063]

[0064] The preparation process of the essence is as follows:

[0065] Step 1: Weigh the raw materials of phase A and mix them evenly, then add deionized water, heat to 80°C and start high-speed homogenization and stirring until completely dissolved, then vacuum heat preservation and defoaming;

[0066] Step 2: Weigh the raw materials of phase B, mix and heat to 85°C, stir and dissolve, then add phase B to phase A and start high-speed homogenization and emulsification for 5 minutes, then cool to 50°C;

[0067] Step 3: Preheat to dissolve phase C, then add phase AB and continue homogenizing for 1 minute;

[0068] Step 4: Cool down to below 40℃, add Phase D ingredients into the main pot and stir at medium speed for 1 minute until evenly mixed, then filter out the material.

[0069] Performance Testing:

[0070] 1. In vitro antibacterial ability test

[0071] (Application example 1 corresponds to embodiment 3, application example 2 corresponds to corresponding example 1, application example 3 corresponds to corresponding example 4, application example 4 corresponds to corresponding example 6, application example 5 corresponds to corresponding example 8, application example 6 corresponds to corresponding example 9, application example 7 corresponds to corresponding example 13)

[0072] Reference Standard - "WS / T 650-2019 Evaluation Methods for Antibacterial and Antibacterial Effects" 5.1.1 Suspension Quantitative Antibacterial Test

[0073] Operation method: Take a 24-hour fresh slant culture of Propionibacterium acnes (ATCC11827) and wash it with PBS, dilute it with PBS to about 5.0×10CFU / mL~4.5×10CFU / mL bacterial suspension for use. Take a sterile test tube, add 5.0mL of sample first, place it in a 20℃±1℃ water bath for 5 minutes, then add 0.1mL of test bacterial suspension, mix quickly and start timing immediately. After the Propionibacterium acnes (ATCC11827) and sample interact for 8 hours, take 1.0mL of the mixture of Propionibacterium acnes (ATCC11827) and sample and inoculate 2 plates respectively, and pour the culture medium. When the amount of bacteria cannot be counted, make a 10-fold serial dilution with PBS, select the appropriate dilution and take 1.0mL to inoculate 2 plates respectively, and count the live bacteria culture. At the same time, use PBS instead of the sample to perform a parallel test as a positive control. The number of colonies recovered in the positive control was between 1.0×10 CFU / mL and 9.0×10 CFU / mL. PBS and culture medium from the same batch were used as negative controls. All test samples and control samples were cultured at 36℃±1℃, and the bacterial propagules were cultured for 48 hours to observe the final results; Candida albicans was cultured for 72 hours to observe the final results. The test was repeated 3 times, and the inhibition rate was calculated. Result judgment: If the inhibition rate is ≥50% to 90%, it is judged to have an antibacterial effect; if the inhibition rate is ≥90%, it is judged to have a strong antibacterial effect.

[0074] Calculation of antibacterial rate: X = (A0-A1) / A0*100%

[0075] Where:

[0076] X—inhibition rate, %;

[0077] A0—the amount of bacteria recovered in the positive control group, in CFU / mL;

[0078] A1—The amount of bacteria recovered in the test group, in CFU / mL.

[0079] The results are shown in Table 4:

[0080] Table 4

[0081] sample Antibacterial rate / % Inhibits the effect of Propionibacterium acnes Application Example 1 99.9 Strong antibacterial Application Example 2 89.6 Has antibacterial effect Application Example 3 88.2 Has antibacterial effect Application Example 4 90.5 Has antibacterial effect Application Example 5 52.5 Has antibacterial effect Application Example 6 51.8 Has antibacterial effect Application Example 7 45.6 No antibacterial effect

[0082] Result analysis: It can be seen from the observation of application examples 5 and 6 that when the composition lacks any of the antibacterial components, octanoyl glycine or polylysine, the antibacterial ability of the composition has declined significantly, and when both are missing, the antibacterial rate of application example 7 has further declined, and failed to show the inhibitory effect on Propionibacterium acnes;

[0083] At the same time, it can be seen from application examples 2-4 that when any one of dipeptide-15, 10-hydroxydecanoic acid and bisabolol is missing from the composition, the antibacterial rate of the composition also declines to a certain extent. Although dipeptide-15, 10-hydroxydecanoic acid and bisabolol have not been recorded to have obvious antibacterial effects in the traditional sense, it can be seen from the above results that dipeptide-15, 10-hydroxydecanoic acid and bisabolol can enhance the antibacterial effects of acylglycine and polylysine to a certain extent.

[0084] 2. In vitro oil control test (5α-reductase inhibition rate test)

[0085] (Application Example 8 corresponds to Example 3, Application Example 9 corresponds to Example 2, Application Example 10 corresponds to Example 4, Application Example 11 corresponds to Example 5, Application Example 12 corresponds to Example 7, Application Example 13 corresponds to Example 8, Application Example 14 corresponds to Example 11)

[0086] Operation method: Dissect the mouse liver, add Tris buffer at a mass ratio of 1:5 and homogenize at 0℃, centrifuge at 4℃, 10000g for 15 minutes, centrifuge the supernatant at 0℃, 55000g for 1 hour, take the supernatant, and place it at 4℃ for later use. Add reagents and samples according to the reaction system, the final concentration of the sample reaction is 1% dilution of the original sample (i.e. 0.01mL / mL), make 5 2-fold gradient dilutions of the positive control finasteride, and repeat each dilution 4 times. After incubation at 37℃ for 60 minutes, measure the absorbance decrease value ⊿A.

[0087] Inhibition rate (%) = (blank ⊿A-sample ⊿A) / blank ⊿A*100

[0088] The results are shown in Table 5:

[0089] Table 5

[0090]

[0091] Result analysis: It can be seen from the observation of application examples 8, 10, 11, and 14 that when the composition lacks both 10-hydroxydecanoic acid and yeast / zinc fermentation product, the 5α-reductase inhibition rate of application example 14 is reduced by about 32% relative to that of application example 8, and when the composition lacks any one of 10-hydroxydecanoic acid and yeast / zinc fermentation product, the 5α-reductase inhibition rate of the compositions of application examples 10 and 11 is increased by about 10% and 13% relative to that of application example 14, respectively. It can be seen that 10-hydroxydecanoic acid and yeast / zinc fermentation product produce a synergistic effect. The reason is that if application example 8 is just the effect of the simple superposition of 10-hydroxydecanoic acid and yeast / zinc fermentation product on the 5α-reductase inhibition rate, then the 5α-reductase inhibition rate of application example 8 should be around 30%, but the 5α-reductase inhibition rate of application example 8 reaches 38.21%, which is far more than the effect of simple superposition.

[0092] Secondly, it can be seen from Application Examples 9, 12 and 13 that when the composition lacks any one of olive leaf extract, tetrahydromethylpyrimidine carboxylic acid and capryloylglycine, the 5α-reductase inhibition rate of Application Examples 9, 12 and 13 also declines to a certain extent. Although they do not show good 5α-reductase inhibition ability in the traditional sense, it can be seen from the above results that when olive leaf extract, tetrahydromethylpyrimidine carboxylic acid and capryloylglycine are used together with 10-hydroxydecanoic acid and yeast / zinc fermentation product, olive leaf extract, tetrahydromethylpyrimidine carboxylic acid and capryloylglycine can enhance the 5α-reductase inhibition ability of the composition to a certain extent.

[0093] 3. In vitro 1L-1α inhibition test (anti-keratinization pathway)

[0094] (Application Example 15 corresponds to Example 3, Application Example 16 corresponds to Example 1, Application Example 17 corresponds to Example 2, Application Example 18 corresponds to Example 3, Application Example 19 corresponds to Example 10)

[0095] Method: Human keratinocytes were cultured in 96-well plates for 24 hours. The cells were then treated with 0.1 μM dexamethasone or 0.01% diluted samples and incubated at 37°C and 5% carbon dioxide for 24 hours. After incubation, the cells were re-treated with 0.01% diluted samples in a medium containing the pro-inflammatory drug PMA (final concentration of 0.1 μg / ml). A control group without PMA was also performed. All treatments were repeated three times. After 48 hours of culture, the culture medium was collected, and IL-1α analysis was performed after measuring IL-1α from the collected culture medium using an ELISA kit. The cell viability was evaluated using the MTT method on the cell layer.

[0096] The results are shown in Table 6:

[0097] Table 6

[0098]

[0099] Result analysis: It can be seen from the observation of application examples 16-18 that when any of dipeptide-15, olive leaf extract, and acetyl glucosamine is missing from the composition, the 1L-1α down-regulation rate of application examples 16-18 is significantly reduced, indicating that the inhibitory ability of the composition on 1L-1α is reduced. At the same time, it can be seen from the observation of application example 19 that it is reduced by about 25% relative to application example 15. When any of dipeptide-15, olive leaf extract, and acetyl glucosamine is missing from the composition, application examples 16-18 are relatively Example 19 increased by about 10%, 6%, and 5% respectively, while in fact, Application Example 15 increased by about 25% relative to Application Example 19. It can be seen that the improvement in the inhibitory ability of 1L-1α in Application Example 15 relative to Application Example 19 far exceeds the inhibitory ability improvement effect of 1L-1α produced by the superposition of dipeptide-15, olive leaf extract, and acetyl glucosamine. Therefore, we believe that the reason why Application Example 15 achieved such a high 1L-1α downregulation rate is the result of the synergistic effect of dipeptide-15, olive leaf extract, and acetyl glucosamine.

[0100] 4. In vitro anti-inflammatory test (hyaluronidase inhibition rate)

[0101] (Application Example 20 corresponds to Example 3, Application Example 21 corresponds to Example 6, Application Example 22 corresponds to Example 7, Application Example 23 corresponds to Example 12)

[0102] Test principle: Hyaluronidase is an endogenous hexosaminidase that causes the decomposition of macromolecular hyaluronic acid. Under the action of hyaluronidase, macromolecular hyaluronic acid decomposes to produce N-acetylglucosamine. Hyaluronidase can hydrolyze the 1,4-glycosidic bond between β-N-acetylglucosamine and D-glucuronic acid in hyaluronic acid to obtain β-N-acetylglucosamine. β-N-acetylglucosamine can condense with acetylacetone under alkaline conditions to generate a chromogen 2-methyl-3-acetylpyrrole derivative, and the chromogen and dimethylaminobenzaldehyde produce color in concentrated hydrochloric acid-ethanol. Therefore, measuring the content of N-acetylglucosamine in the reaction system can indirectly reflect the activity of hyaluronidase. Hyaluronidase has a strong correlation with inflammation, so the inhibition of hyaluronidase activity is used as an indicator to study anti-inflammatory effects.

[0103] Test steps: (1) Preserve CaCl solution and hyaluronidase solution at 37°C for 20 min, then add sample solution;

[0104] (2) Add sodium hyaluronate solution and keep at 37°C for 30 min;

[0105] (3) adding NaOH solution and acetylacetone solution, heating in a boiling water bath, and cooling with ice water;

[0106] (4) Add Ehrlich reagent and let stand for 20 min for color development;

[0107] (5) Measure the absorbance at 547 nm using a spectrophotometer;

[0108] Inhibition rate calculation: Hyaluronic acid inhibition rate = [(AB)-(CD)] / (AB) x 100%

[0109] Where, A——absorbance value of control solution;

[0110] B——Absorbance value of blank control solution;

[0111] C——absorbance value of sample solution;

[0112] D——Absorbance value of sample blank solution.

[0113] The test results are shown in Table 7:

[0114] Table 7

[0115] sample Hyaluronidase inhibition rate (%) Application Example 20 84.0 Application Example 21 59.3 Application Example 22 62.6 Application Example 23 46.5

[0116] Analysis of results: It can be seen from application examples 21 and 22 that when any of bisabolol and tetrahydromethylpyrimidine carboxylic acid is missing in the composition, the hyaluronidase inhibition rate of application examples 21 and 22 is significantly reduced, indicating that the hyaluronidase inhibition ability of the composition is reduced. At the same time, it can be seen from application example 23 that it is reduced by about 37.5% relative to application example 20. When any of bisabolol and tetrahydromethylpyrimidine carboxylic acid is missing in the composition, application examples 21 and 22 are increased by about 13% and 16% relative to application example 19, respectively. In fact, application example 20 is increased by about 37.5% relative to application example 23. It can be seen that the improvement in the hyaluronidase inhibition ability of application example 15 relative to application example 23 far exceeds the hyaluronidase inhibition ability improvement effect caused by the superposition of bisabolol and tetrahydromethylpyrimidine carboxylic acid. Therefore, we believe that the reason why application example 20 achieves such a high hyaluronidase inhibition rate is the result of the synergistic effect of bisabolol and tetrahydromethylpyrimidine carboxylic acid.

[0117] Safety test of application examples / comparative examples - skin occlusion patch test

[0118] Test method - refer to the "Cosmetic Safety Technical Specifications 2015" version - Skin occlusion patch test

[0119] Test steps: Select volunteers aged 18-60 who meet the test requirements as subjects, and exclude unqualified or unsuitable volunteers. Fill out the informed consent form before the test.

[0120] (1) Cleaning

[0121] Gently wipe the test area with a dry soft tissue.

[0122] (2) Patch tester

[0123] According to the preset sample position, stick the corresponding numbered patch tester on the corresponding test area. Quickly stick the patch tester from top to bottom to the corresponding position, gently press and smooth it, and apply it on the skin for 24 hours.

[0124] (3) Spot removal tester

[0125] After applying the patch tester for 24 hours, the guide will take the volunteer to the inspection area, where the tester will remove the patch tester and mark it with a skin marker.

[0126] (4) On-site evaluation and recording of results: 0.5 h after removing the patch (after the indentation disappears), record the results of visual observation in a paper form. After the test is completed, the test personnel will organize the data into an Excel spreadsheet for statistical analysis.

[0127] Skin reactions were observed according to the skin reaction grading standard of skin occlusion patch test in Table 1, and the results were recorded. The scoring standard is shown in Table 8:

[0128] Table 8

[0129]

[0130] During the test, a total of 31 volunteers were enrolled. The observation results 30 minutes after the removal of the spot tester are shown in Table 9:

[0131] Table 9

[0132] Sample name Level 1 reaction Second level reaction Grade 3 reaction Grade 4 reaction Application Example 1 0 0 0 0 Application Example 2 0 0 0 0 Application Example 3 0 0 0 0 Application Example 4 0 0 0 0 Application Example 5 0 0 0 0 Application Example 6 0 0 0 0 Application Example 7 0 0 0 0 Application Example 8 0 0 0 0 Application Example 9 0 0 0 0 Application Example 10 0 0 0 0 Application Example 11 0 0 0 0 Application Example 12 0 0 0 0 Application Example 13 0 0 0 0 Application Example 14 0 0 0 0

[0133] 5. Acne Removal Efficacy Test on Human Body - Visia CR Photo Evaluation

[0134] (1) Subject screening: healthy males or females aged between 18 and 35 years old; those who have not participated in other clinical trials in the past month; those who have not used acne-removing products or received acne-removing treatments (oral hormone drugs, injections, lasers, and other plastic surgeries) in the past month. Women who have acne during menstruation and those who occasionally have acne are excluded; only those with grade I or II facial acne, more than 6 pimples, accompanied by obvious redness and swelling, and a relatively even distribution of pimples on the left and right sides of the face, who have been troubled by repeated acne growth for a long time, are retained; a total of 16 people were screened;

[0135] (2) Trial steps: On the day of the visit, the subjects washed their faces without applying any skin care products. They sat quietly in a constant temperature and humidity room at a temperature of 21±1°C and a humidity of 50±10% for 20 minutes, and used Visia-CR to take photos of the left, right and middle faces of the face and used them as D0 data; then the subjects used the acne essence-application example 3 sample on the right face three times a day in the morning, noon and evening, and did not use it on the left face. Comparison was made on the 3rd, 7th, 14th, 21st and 28th days of use. The left, right and middle faces of the face were photographed with the Visia-CR device. The subjects' skin erythema index EI value, pore number, porphyrin, skin acne count, skin stinging score (subjective evaluation), use mildness (subjective evaluation), and use effect evaluation (subjective evaluation) were analyzed for improvement, and the results were described as a percentage decrease (%) or a Likert five-level scale for subjective evaluation.

[0136] The results are shown in Table 10:

[0137] Table 10

[0138]

Claims

1. An acid-free acne-removing composition, characterized in that: By mass fraction, it specifically includes the following components:

2. The acid-free anti-acne composition according to claim 1, characterized in that: By mass fraction, it specifically includes the following components:

3. The acid-free anti-acne composition according to claim 1, characterized in that: The polyol is selected from at least one of glycerol, butylene glycol, dipropylene glycol, 1,3-propanediol and methylpropylene glycol.

4. Use of the acid-free anti-acne composition according to any one of claims 1 to 3 as an anti-acne active ingredient in daily chemical products.

5. A daily chemical product, characterized in that: Contains 8-12%wt of the acid-free anti-acne composition according to any one of claims 1-3.

6. The daily chemical product according to claim 5, characterized in that: The daily chemical product is lotion, essence or facial mask.

7. The daily chemical product according to claim 5, characterized in that: The dosage form of the daily chemical product is spray, lotion or cream.

Images