A soothing composition, a firming anti-wrinkle composition and uses thereof, firming anti-wrinkle skin care

Abstract

This invention belongs to the field of cosmetic technology and discloses a soothing composition, a firming and anti-wrinkle composition and their uses, and a firming and anti-wrinkle skin care product. The soothing composition comprises the following components by weight percentage: milk thistle fruit extract: 10wt%-20wt%; rosewood bark extract: 5wt%-10wt%; swertia bud extract: 1wt%-2wt%; squalane: 68wt%-84wt%. The soothing composition combines the three extracts simultaneously, which has a significant synergistic effect in increasing the soothing effect of the composition.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology and discloses a soothing composition and its uses, as well as skin care products. Background Technology

[0002] With the fast pace of life and increased consumer attention to skincare, sensitive skin problems are becoming increasingly prominent. Sensitive skin often manifests as subjective symptoms such as burning, stinging, itching, and tightness when stimulated by physical, chemical, or psychological factors, with or without objective signs such as erythema, scaling, and telangiectasia. Damaged skin barrier function not only leads to immediate symptoms such as dryness, redness, and itching, but also triggers chronic inflammatory responses, accelerating the skin aging process.

[0003] While numerous cosmetic products on the market claim to soothe the skin, several technological bottlenecks remain. On one hand, most products employ a single active ingredient system, resulting in limited efficacy. For example, collagen products primarily rely on the physical film-forming effect of large-molecule collagen; however, their transdermal absorption rate is low, generally below 8%, and they struggle to activate extracellular matrix regeneration. Although Centella asiatica extract possesses anti-inflammatory properties, its triterpenoid components (such as asiaticoside) are easily affected by polyol solvent systems in conventional formulations, leading to decreased bioavailability. Hydrolyzed gels, due to their small molecular weight, while capable of penetrating the stratum corneum when used alone, lack long-lasting moisturizing capabilities. On the other hand, the preservative systems of these products also present problems. Many products on the market rely on phenoxyethanol and parabens as preservatives, which can disrupt the skin's microbiome balance and exacerbate the immune stress response of sensitive skin. In addition, in some products with special functional ingredients, such as cosmetics that add retinoids to achieve anti-aging effects, even if the stability of retinoids is improved and the irritation is reduced through encapsulation and sustained-release technologies, the formula still poses a significant risk of skin irritation when the amount of retinoids added is increased and the product is used continuously.

[0004] Meanwhile, some products claiming cooling and soothing effects also have shortcomings. For example, menthol, a commonly used cooling ingredient, while providing a strong cooling sensation, is highly volatile and its cooling effect is short-lived. Its effectiveness diminishes rapidly with prolonged use and storage. Increasing the dosage to prolong the cooling effect not only yields insignificant results but can also irritate the skin, causing localized pain or even a burning sensation. Furthermore, menthol and its derivatives have poor solubility, often requiring high-concentration ethanol solutions for dissolution. In high-temperature or low-temperature environments with high water content, this can easily lead to crystallization, water-oil separation, and other issues, resulting in unstable product quality.

[0005] Therefore, the technical problem that this invention needs to solve is: how to provide a new, stable and efficient soothing composition. Summary of the Invention

[0006] The purpose of this invention is to provide a soothing composition containing milk thistle fruit extract, rosewood bark extract, and swertia spp. extract. The combination of these three components has a significant synergistic effect in enhancing the soothing effect of the composition.

[0007] In addition, the present invention also provides the uses of the soothing composition and skin care products.

[0008] To achieve the above objectives, the present invention provides the following technical solution: A soothing composition comprising, by weight percentage: Milk thistle fruit extract: 10wt%-20wt%; Dalbergia odorifera bark extract: 5wt%-10wt%; Indian swert sprout extract: 1wt%-2wt%; Squalane: 68wt%-84wt%.

[0009] Furthermore, the present invention discloses a firming and anti-wrinkle composition comprising: 68wt%-84wt% of squalane and 16wt%-32wt% of a functional ingredient, wherein the functional ingredient is at least one selected from milk thistle fruit extract, palm bark extract, and swertia spp. extract.

[0010] Preferably, the functional ingredient is swert extract, palm bark extract, a combination of palm bark extract and swert extract, or a combination of palm bark extract and milk thistle fruit extract.

[0011] Preferably, when the functional component is a combination of Dalbergia odorifera bark extract and Swertia gigantea extract, the mass ratio of Dalbergia odorifera bark extract to Swertia gigantea extract is 18:6.

[0012] Preferably, when the functional component is a combination of palm bark extract and milk thistle fruit extract, the mass ratio of palm bark extract to milk thistle fruit extract is 16.5:7.5.

[0013] Furthermore, this invention discloses the use of the soothing composition described above in the preparation of skin care products.

[0014] Furthermore, this invention discloses the use of the firming and anti-wrinkle composition described above in the preparation of skin care products.

[0015] In addition, the present invention discloses a skin care product containing the soothing composition described above.

[0016] Finally, the present invention discloses a skin care product containing the firming and anti-wrinkle composition as described above.

[0017] Preferably, the content of the composition in the skin care product is 0.5wt%-5wt%.

[0018] More preferably, the dosage form of the skin care product is one of lotion, cream, spray, serum, essential oil, or toner.

[0019] Compared with the prior art, the beneficial effects of the present invention are: The soothing composition of the present invention contains milk thistle fruit extract, palm bark extract and swertia bud extract. The simultaneous use of these three components has a significant synergistic effect in increasing the soothing effect of the composition. Attached Figure Description

[0020] Figure 1 The image shows the cytotoxicity test results of a face cream used in an application example. Figure 2 The graph shows the average type I collagen concentration of the face cream used in the application example. Detailed Implementation

[0021] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0022] Product Information: Milk thistle fruit extract: Milk thistle fruit extract (100%) Guangzhou Qianjinghui Biotechnology Co., Ltd.; Dalbergia odorifera bark extract: Dalbergia odorifera bark extract (100%) Guangzhou Maisihui Biotechnology Co., Ltd.; Indian swert extract: maltodextrin (98%), Indian swert extract (2%), Guangzhou Yihe Trading Co., Ltd.; Squalane: Squalane (100%) Guangzhou Jinhang Biotechnology Co., Ltd.

[0023] It should be noted that the technical solution of this application can be achieved by purchasing other commercially available raw materials, and is not limited to the aforementioned raw material suppliers.

[0024] The formulations of the compositions in each embodiment and comparative example are shown in Table 1; the preparation methods are as follows: Mix squalane with milk thistle fruit extract and palm bark extract according to the specified weight parts. Then slowly add the mixture to 4 parts of emulsifier sucrose laurate to form a gel-like mixture. Finally, add swerts of swertia extract to the gel mixture and stir until well combined.

[0025] It should be noted that the emulsifier sucrose laurate only serves to mix the composition and is not an active ingredient. Those skilled in the art can choose other commonly used emulsifiers as needed, which will not have a significant impact on the efficacy of the composition.

[0026] Table 1. Combination Formula (parts by weight) Example 1 15 7 2 76 Example 2 10 5 1 84 Example 3 20 10 2 68 Comparative Example 1 0 18 6 76 Comparative Example 2 21 0 3 76 Comparative Example 3 16.5 7.5 0 76 Comparative Example 4 24 0 0 76 Comparative Example 5 0 24 0 76 Comparative Example 6 0 0 24 76

[0027] Performance testing Hyaluronidase inhibition rate I. Experimental Principle Hyaluronic acid, as an important natural moisturizing substance, plays a vital role in maintaining the volume of the extracellular matrix and normal skin physiological functions. However, hyaluronic acid can be specifically broken down by hyaluronidase, and its products promote skin inflammation, leading to increased histamine production in the body. Furthermore, hyaluronidase expression is also associated with most IgE-mediated type I and T-cell-mediated type IV hypersensitivity reactions. Inhibiting hyaluronidase activity can, to some extent, help suppress the development of skin inflammation and allergic reactions, and improve the irritated state of the skin caused by inflammation and allergic reactions. Therefore, inhibition of hyaluronidase activity is often used as an important indicator for evaluating the soothing and moisturizing effects of products.

[0028] Hyaluronidase breaks the glycosidic bond between N-acetylglucosamine and D-glucuronic acid in the hyaluronic acid (HA) sugar chain. The resulting reducing sugar can reduce DNS to an amino compound under alkaline conditions. After boiling in a water bath to allow for sufficient color development, its absorbance can be measured at 540 nm, thereby calculating the hyaluronidase activity.

[0029] II. Materials and Methods 1. Instruments and equipment: microbalance, water bath, microplate reader.

[0030] 2. Reagents and materials: Hyaluronidase, hyaluronic acid; Positive control: Dexamethasone, DNS reagent.

[0031] 3. Operation process 3.1 Test Operation Sample groups: Examples 1-3 and Comparative Examples 1-6, without dilution.

[0032] Referring to the sample addition grouping list in Table 2, the experiment was conducted using 2mL centrifuge tubes, with 3 parallel tubes in each group.

[0033] Table 2 Sample Addition Grouping List

[0034] After the test tube has cooled, shake well and add 150 μL of the reaction solution to a 96-well plate. Measure the absorbance at OD540 nm using an ELISA reader.

[0035] 3.2 Calculation of Results The inhibition rate of hyaluronidase activity was calculated according to Formula I: Formula I: ; In the formula: T: Absorbance of the sample group, i.e., the absorbance of the solution after the sample inhibits the reaction between hyaluronidase and hyaluronic acid; T0: Background absorbance of the sample; C: Absorbance value of the blank control group, that is, the absorbance value of the reaction between hyaluronidase and hyaluronic acid when no sample is added; C0: Absorbance of the solvent background group.

[0036] The results of the hyaluronidase activity inhibition rate are shown in Table 3.

[0037] Table 3. Data on hyaluronidase inhibition rate Example 1 77.52 Example 2 74.48 Example 3 79.28 Comparative Example 1 66.92 Comparative Example 2 55.73 Comparative Example 3 60.46 Comparative Example 4 51.61 Comparative Example 5 70.32 Comparative Example 6 64.57 Positive control 51.32

[0038] As shown in Table 3, Examples 1-3 have a high hyaluronidase inhibition rate, indicating that the compositions within the dosage range of the present invention all have a significant hyaluronidase inhibition effect and a good soothing effect.

[0039] Data from Examples 1-6 show that when the three components are used alone with squalane, the most effective component is the rosewood bark extract, which has a hyaluronidase inhibition rate of 70.32%. When the three components are combined in any two pairs with squalane, the effect is a simple summation of the effects of each component used alone with squalane. Comparing the data from Examples 1 and 1-6, it can be seen that when the three components are used in combination with squalane, they have a synergistic effect in improving the hyaluronidase inhibition rate of the composition. The content of the Dalbergia odorifera bark extract in Example 1 is much lower than that in Comparative Example 5, but when it is used in combination with milk thistle fruit extract and swertia bud extract, its hyaluronidase inhibition rate is as high as 77.52%, which is about 7% higher than that in Comparative Example 5. The amount of milk thistle fruit extract and Dalbergia odorifera bark extract in Comparative Example 3 is similar to that in Example 1, but it lacks swertia bud extract, and its hyaluronidase inhibition rate is only 60.46%, which is about 17% lower than that in Example 1.

[0040] The above analysis leads to the conclusion that combining milk thistle fruit extract, rosewood bark extract, and swertia bud extract within the dosage range of this invention with squalane has a significant synergistic effect in enhancing the hyaluronidase inhibition rate of the composition. Furthermore, all three components are indispensable for the composition to exert its highly effective soothing effect.

[0041] DPPH free radical scavenging rate I. Principle 1,1-Diphenyl-2-trinitrophenylhydrazine (DPPH) is a stable, long-lived free radical. Its ethanol solution is deep purple and exhibits strong absorption around 517 nm. In the presence of a free radical scavenger, the light absorption of the DPPH ethanol solution decreases due to single electron pairing. The degree of fading of the DPPH ethanol solution is linearly related to the number of electrons it accepts, which can be used to evaluate the test sample's ability to scavenge free radicals.

[0042] II. Instruments and Equipment: Microbalance, Microplate reader.

[0043] III. Reagents and materials: 95% ethanol, 1,1-diphenyl-2-trinitrophenylhydrazine, positive control: water-soluble vitamin E.

[0044] IV. Operating Procedures 4.1 Sample preparation: Weigh a quantitative amount of sample. Water-soluble test substances are diluted with water to prepare multi-concentration samples. Oil-soluble test substances are diluted with 95% ethanol to prepare multi-concentration samples. Insoluble samples are prepared in units of mass and volume. During the reaction, shake and stir thoroughly to mix well and then let stand.

[0045] 4.2 Test Operation DPPH working solution: Dissolve DPPH reagent in 95% ethanol to prepare a solution of 0.12 mg / mL. Prepare in the dark immediately before use. Positive control: Water-soluble vitamin E was prepared into a stock solution of 0.08 mg / mL using 95% ethanol, and then diluted 1 / 2 times to create a series of concentration gradients to verify the test system. The concentration settings should include 0.01, 0.02, 0.04, and 0.08 mg / mL.

[0046] Referring to the sample addition grouping list in Table 4, experiments were conducted using 96-well microplates. Sample groups (T) for Examples 1-3 and Comparative Examples 1-6, a sample background group (T0), a negative control group (C), and a solvent background group (C0) were established. The positive control group was treated the same as the sample groups. Three replicates were set up for each group. Reagents were added sequentially using a multi-channel pipette to ensure consistent reaction times.

[0047] Table 4 Sample Addition Grouping List Sample solution (μL) 50 50 - - Water or 95% ethanol solvent (μL) 100 100 150 150 DPPH ethanol solution (μL) 50 - 50 - 95% ethanol (μL) - 50 - 50

[0048] After reacting in the dark for 5 minutes, the absorbance was immediately measured using an ELISA reader with the following parameters: slow shaking for 10 seconds, and OD517mm absorbance measured.

[0049] 4.3 Result Calculation The DPPH free radical scavenging rate is calculated according to Formula II: Formula II: ; In the formula: T: Absorbance of the sample group, i.e., the absorbance of the solution after the sample reacts with DPPH; T0: Background absorbance of the sample; C: Absorbance value of the blank control group, i.e., the absorbance value of the DPPH solution without the addition of sample; C0: Absorbance of the solvent background group.

[0050] The DPPH free radical scavenging results are shown in Table 5.

[0051] Table 5 DPPH Free Radical Scavenging Rate Data Example 1 42.94 Example 2 49.63 Example 3 48.60 Comparative Example 1 57.73 Comparative Example 2 20.65 Comparative Example 3 63.60 Comparative Example 4 30.67 Comparative Example 5 40.30 Comparative Example 6 51.34 Positive control 73.80

[0052] The data from Comparative Examples 4-6 show that when the three components are used alone with squalane, the best DPPH free radical scavenging rate is achieved by *Swertia javanica* extract, followed by *Palmaria rubra* bark extract, and the worst is by *Silybum marianum* fruit extract.

[0053] Data from Comparative Examples 1-3 show that the simultaneous use of *Dalbergia odorifera* bark extract and *Swertia argyrophylla* extract, or the simultaneous use of *Silybum marianum* fruit extract and *Dalbergia odorifera* bark extract, has a synergistic effect in enhancing the DPPH free radical scavenging rate of the composition. However, the combined use of *Silybum marianum* fruit extract and *Swertia argyrophylla* extract exhibits an antagonistic effect, resulting in a decrease in the DPPH free radical scavenging rate of the composition. Further data from Examples 1-3 indicate that when the three components are used together, *Dalbergia odorifera* bark extract can effectively reduce the antagonistic effect of *Silybum marianum* fruit extract and *Swertia argyrophylla* extract.

[0054] Application examples This invention discloses a face cream containing the composition of Example 1, the formulation of which is shown in Table 6, and the preparation method is as follows; 1. Aqueous phase: Add phase A to the emulsification pot and stir to 85℃ (stirring frequency 30Hz). Homogenize for 2 minutes until there are no clumps (homogenization frequency 30Hz). Set aside.

[0055] 2. Oil phase: Add phase B to the oil phase pot, heat to 85℃, and stir until completely dissolved (stirring frequency 30HZ), then set aside.

[0056] 3. When phases A and B reach the specified temperature of 84℃, phase B is drawn into phase A and homogenized for 3 minutes (homogenization frequency 30Hz). Phase C is added and homogenized for 2 minutes (homogenization frequency 30Hz). The mixture is kept warm for 30 minutes to remove bubbles. Then, phase D is added (no further heat preservation is required after adding phase D). The mixture is stirred and cooled (stirring frequency 30Hz).

[0057] 4. After cooling to 60-65℃, add E1 and E2 phases and homogenize for 2 minutes (homogenization frequency 30Hz), then stir evenly (stirring frequency 30Hz).

[0058] 5. Cool down to below 45℃, add F and pre-dissolved G phase in sequence and stir evenly (stirring frequency 30Hz). After passing the inspection, filter the material through a 150-mesh filter cloth.

[0059] Table 6 Face Cream Recipe Table

[0060] The obtained face cream was subjected to the following tests. Multiple skin irritation tests I. Materials and Methods 1. Test substance: The face cream prepared by the application example was used directly without any treatment.

[0061] 2. Laboratory animals and their housing: Laboratory animals: 4 ordinary-grade New Zealand rabbits, female, non-pregnant, never given birth, weighing 2.12-2.22 kg. The skin was intact and without abnormalities upon examination. The laboratory animals were purchased from Huadong Xinhua Laboratory Animal Breeding Farm, Huadu District, Guangzhou City. Laboratory animal production license number: SCXK (Guangdong) 2024-0023, quality certificate number: 44007600013444.

[0062] Breeding environment: temperature 18-26℃, relative humidity 30-70%, laboratory animal use license number: SYXK (Guangdong) 2022-0198.

[0063] Feed: Purchased from Qingdao Kangda Aibo Biotechnology Co., Ltd., Certificate No.: 3708702500056994.

[0064] 3. Test method: Chapter 6 (4) of the "Cosmetic Safety Technical Specifications" (2015 edition).

[0065] 3.1 Application Method: Before the experiment, the hair on both sides of the spine on the back of the experimental animal was shaved, with a shaved area of ​​approximately 3cm × 3cm on each side, and an application area of ​​approximately 2.5cm × 2.5cm. 0.5mL of the test substance was applied to one side of the skin, with the other side serving as a blank control. Application was repeated once daily for 14 consecutive days. Starting from the second day, the hair was shaved before each application, and any residual test substance was removed with water. Results were observed one hour later. The control and test areas were treated in the same way.

[0066] 3.2 Animal observation: Skin reaction was scored according to the skin irritation response scoring criteria. The average score per animal per day was calculated using the following formula. The intensity of skin irritation was determined according to the skin irritation intensity grading standard: .

[0067] II. Test results, see Table 7.

[0068] Table 7 Results of multiple skin irritation tests

[0069] As can be seen from the data in Table 7, the face cream containing the composition of Example 1 prepared by the application example showed no skin irritation to rabbits after repeated use.

[0070] Type I collagen content assay (human fibroblasts) I. Principle Normal mammalian cells cultured in vitro continuously divide and proliferate. Toxic substances, regardless of their site of action or mechanism of action, will interfere with the cell division and proliferation process, leading to a decrease in cell growth rate and number.

[0071] During their proliferation and division, cultured dermal fibroblasts secrete substances such as collagen. The efficacy of a test substance in firming and anti-wrinkle cell function can be evaluated by measuring the expression of related proteins after administration. This document uses human fibroblasts (HDF) and a human enzyme-linked immunosorbent assay (ELISA) kit to detect type I collagen (Collagen I). The analyte specifically binds to the antibody coated on the ELISA plate, and then to the substrate-labeled antibody. The substrate is catalyzed by the enzyme to generate a colored product. The concentration of the analyte is positively correlated with the intensity of the colored product. The concentration of the analyte is calculated by measuring the optical density (OD value) at 450 nm using an ELISA reader.

[0072] II. Instruments and Equipment: Microbalance, CO2 incubator, microplate reader, water bath, biosafety cabinet.

[0073] III. Reagents and Materials CCK-8 reagent; DMEM, 4.5 g / L, purchased from Glucose; FBS; 0.25% Trypsin-EDTA digestive enzyme; Col I ELISA kit; human fibroblasts.

[0074] IV. Operating Procedures 4.1 Test substance pretreatment: The original sample solution was sterilized by filtration using a 0.22 μm filter, and then diluted to the required concentration using culture medium.

[0075] 4.2 Cytotoxicity / Proliferation Assay: After cell counting, the cells were diluted to the required concentration and added to 96-well microplates. After 24 hours of plating, when the cell confluence rate was 40-60%, the test substance was added using a multipipeline to achieve the set concentration. A solvent control was included, and the test wells were set up with parallel runs (n ​​≥ 4). 48 hours after sample addition, the waste liquid was removed, and DMEM containing 10% CCK-8 was added. After incubation for 1-1.5 hours, the cells were read using a microplate reader with an OD of 450 nm.

[0076] 4.3 Type I Collagen Concentration Assay: After cell counting, the cells were diluted to the required concentration and added to a 96-well microplate. After 24 hours of plating, when the cell confluence rate was 40-60%, the test concentration of the reagent was added and incubated for another 24 hours. 200 μL of cell culture supernatant was collected from each well into a 1.5 mL sterile centrifuge tube, and the assay was performed according to the instructions of the Col I ELISA kit.

[0077] 4.4 Result Calculation 4.4.1 Calculation of cell viability in the toxicity test of the test substance: ; In the formula: Cn - contains the test substance and has cells; C0: Solvent control, no test substance, cells present; VCb: Solvent blank control, no test substance, no cells.

[0078] The cell viability test results in the toxicity test of the test substance are as follows: Figure 1 As shown in Table 8.

[0079] Table 8. Results of Cytotoxicity Test 0.00 100.00 3.27 0.01 117.80 2.65 0.02 112.53 2.54 0.04 113.21 4.00 0.08 113.32 2.77 0.16 116.47 4.01 0.31 120.20 3.49 0.63 127.73 2.35 1.25 133.44 2.28 2.50 132.62 3.52

[0080] Based on the results in Table 8, a concentration of 2.50 mg / mL was selected for the type I collagen concentration secretion test.

[0081] 4.4.2 Calculation of Average Type I Collagen Concentration: A curve was plotted with absorbance (OD) as the ordinate (Y) and the corresponding concentration of the analyte standard as the abscissa (X). The concentration of the analyte in the sample could be calculated from the standard curve based on its OD value. The final result of the analyte concentration was obtained by taking the average value of the three replicates for each group.

[0082] Reference Table 9 shows the results of the Type I collagen secretion test. Figure 2 .

[0083] Table 9. Results of Average Type I Collagen Concentration BC 53.96 1.14 PC: 300 ng / mL TGF 75.26 0.50 2.5mg / mL face cream 93.79 5.09

[0084] From Table 9 and Figure 2 It can be seen that, compared with the control group, adding 2.5 mg / mL of face cream can significantly increase the average concentration of type I collagen.

[0085] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, it is intended that all variations falling within the meaning and scope of equivalents of the claims be included within the present invention.

Claims

1. A soothing composition, characterized in that, It consists of the following components by mass percentage: Milk thistle fruit extract: 10wt%-20wt%; Dalbergia odorifera bark extract: 5wt%-10wt%; Indian swert sprout extract: 1wt%-2wt%; Squalane: 68wt%-84wt%.

2. Use of the soothing composition as described in claim 1 in the preparation of skin care products.

3. A skincare product, characterized in that, The skincare product contains the soothing composition as described in claim 1.

4. The skincare product according to claim 3, characterized in that, The skincare product is in one of the following forms: lotion, cream, spray, serum, essential oil, or toner.

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