Highly absorbable whitening and freckle-removing skin care product and preparation process thereof

Abstract

The application discloses a high-efficiency absorption whitening and freckle-removing skin care product and a preparation process thereof, and belongs to the technical field of cosmetics. The high-efficiency absorption whitening and freckle-removing skin care product contains the following components in terms of weight parts: 0.1-5 parts of a whitening compound (I), 5-10 parts of cocoyl glucoside, 0.5-1 part of menthol, 5-10 parts of phospholipid, 0.5-1 part of EDTA, 0.1-1 part of hyaluronic acid and 150-200 parts of deionized water. The whitening and freckle-removing skin care product has good antioxidant and tyrosinase enzyme activity inhibiting effects, and can realize the effects of whitening and freckle removal.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, specifically to a highly absorbent whitening and spot-fading skincare product and its preparation process. Background Technology

[0002] Human skin is divided into the epidermis and dermis. The stratum corneum is the outermost layer of the epidermis and is the main barrier for the transdermal transport of most substances. The intercellular lipids in the stratum corneum are mainly composed of ceramides, cholesterol, and fatty acids, existing in a multilayered, sheet-like bimolecular form. Most of the proteins in the stratum corneum are keratin fibers condensed from keratinized cells. Substances with a relative molecular weight greater than 500 do not easily penetrate the stratum corneum.

[0003] Melanocytes are mainly distributed in the epidermis. The raw material required for melanin synthesis is tyrosine. Tyrosine is synthesized into melanin in membrane-bound vesicles under the catalysis of tyrosinase. Melanocytes then transfer melanin to adjacent keratinocytes. The currently accepted melanin formation pathway is: tyrosine-DOPA-DOPAquinone-DOPAchrome-dihydroxyindole-ketoindole melanin. The main whitening methods currently include: 1. Directly inhibiting melanin production. Tyrosinase is the rate-limiting enzyme in melanin synthesis; inhibiting its activity can achieve a whitening effect. For example, phenylethyl resorcinol is commonly used in skincare products from major brands. 2. Antioxidant effects, blocking melanin production by reducing intermediates in the melanin formation process. 3. Inhibiting the transfer of melanin to keratinocytes. 4. Exfoliation of the stratum corneum, accelerating melanin metabolism. 5. Reducing ultraviolet radiation.

[0004] Although existing technologies have clarified the pathways of melanin production and specific whitening methods, the whitening ingredients in whitening skincare products on the market are still mainly phenylethyl resorcinol, arbutin, vitamin C, niacinamide, fruit acids, plant extracts, and peptides. There has been a lack of more effective whitening ingredients, and technological breakthroughs in new products have not been achieved. Therefore, in order to meet the market's demand for whitening skincare, there is an urgent need to provide a whitening and spot-fading skincare product that can be absorbed efficiently. Summary of the Invention

[0005] In order to overcome the shortcomings of the existing technology, the purpose of this invention is to provide a highly absorbent whitening and spot-fading skin care product and its preparation process, which has the technical effects of whitening and spot fading, low irritation, skin toning and moisturizing.

[0006] This invention is achieved through the following technical solution: A skin whitening and spot-fading skin care product, characterized in that, by weight, it comprises the following components: 0.1-5 parts of whitening compound (Ⅰ), 5-10 parts of cocoyl glucoside, 0.5-1 parts of menthol, 5-10 parts of phospholipids, 0.5-1 parts of EDTA, 0.1-1 parts of hyaluronic acid, and 150-200 parts of deionized water, wherein the molecular structure of the whitening compound is as follows: Formula (Ⅰ):

[0007]

[0008] The preparation method of the whitening compound (Ⅰ) includes the following steps:

[0009] S1: Under a nitrogen atmosphere, dissolve 3,4-dihydroxymandelic acid in anhydrous DMF (N,N-dimethylformamide), then add EDC and NHS sequentially, and stir until activation is complete to form an activated 3,4-dihydroxymandelic acid solution for later use.

[0010] S2: Under a nitrogen atmosphere, a DMF solution of 4-aminoresorcinol was slowly added dropwise to the activation solution. The mixture was stirred at room temperature until the reaction was complete. The solution was then purified by column chromatography to obtain the purified whitening compound (Ⅰ).

[0011] The molar ratio of 3,4-dihydroxymandelic acid, EDC and NHS is 1:(1-1.2):(1-1.2), preferably 1:1.2:1.2.

[0012] The molar ratio of 3,4-dihydroxymandelic acid to 4-aminoresorcinol is 1:(1-1.1), preferably 1:1.

[0013] The preferred dosage of the whitening compound (Ⅰ) is 0.1-3 parts.

[0014] The phospholipid is selected from at least one of 1,2-distearyl-sn-glycerol-3-phosphoethanolamine (DSPE), sodium dioleoylphosphatidylserine (DOPS), egg yolk lecithin, soybean lecithin, dioleoyl L-α-phosphatidylethanolamine (DOPE), 1,2-dioleoyl lecithin (DOPC), dipalmitoylphosphatidylcholine (DPPC), and distearylphosphatidylcholine (DSPC).

[0015] Beneficial effects

[0016] This invention provides a skin-whitening and spot-fading skincare product. A novel skin-whitening compound (Ⅰ) is synthesized from 4-aminoresorcinol and 3,4-dihydroxymandelic acid. This compound has a similar chemical structure to phenylethylresorcinol, exhibiting similar biphenyl rings and polyphenolic hydroxyl structures. Phenethylresorcinol is a highly effective skin-whitening ingredient used in skincare and cosmetic products from brands such as L'Oréal, possessing antioxidant and tyrosinase-inhibiting properties. Compared to phenylethylresorcinol, the skin-whitening compound (Ⅰ) of this invention exhibits stronger antioxidant activity and tyrosinase-inhibiting properties, and its water solubility is improved, making it more effective in exerting its skin-whitening effect within skin cells.

[0017] When cocoyl glucoside is used in combination with phospholipids, it can form stable micelles that encapsulate the whitening compound (I), increasing its lipophilicity. Cocoyl glucoside and menthol can disrupt the stratum corneum structure and improve the fluidity of the phospholipid membrane in stratum corneum cells. The synergistic effect of cocoyl glucoside, phospholipids, and menthol significantly enhances the transdermal absorption of whitening ingredients and also provides a toning effect. The addition of EDTA improves product stability and effectively prevents the oxidative degradation of the whitening compound (I), which may be related to EDTA's ability to chelate metal ions.

[0018] The whitening skincare product described in this invention has highly effective whitening and spot-fading, antioxidant, and transdermal absorption effects; it is water-based, non-greasy, and enhances the skin's moisturizing effect. Attached Figure Description

[0019] Figure 1 Synthetic pathway diagram of skin whitening compound (Ⅰ).

[0020] Figure 2 Near-infrared spectrum of whitening compound (I).

[0021] Figure 3 Image of the powder containing the skin whitening compound (Ⅰ).

[0022] Figure 4 Tyrosinase activity inhibition rate graph. Detailed Implementation

[0023] The technical solutions 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.

[0024] In the embodiments, unless otherwise specified, the experimental methods used are conventional methods, and the materials and reagents used are commercially available unless otherwise specified.

[0025] The raw materials used in the examples and comparative examples are described below:

[0026] 4-Aminoresorcinol: purchased from Nanjing Qianyida New Material Technology Co., Ltd.;

[0027] 3,4-Dihydroxymandelic acid: purchased from Shanghai Yuanye Biotechnology Co., Ltd.;

[0028] EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) was purchased from Jier Biochemical (Shanghai) Co., Ltd.

[0029] NHS (N-hydroxysuccinimide): Purchased from Hubei Xinghengkang Chemical Technology Co., Ltd.

[0030] Phospholipid HSPE: purchased from Shenzhen Zhenqiang Biotechnology Co., Ltd.;

[0031] Phenethyl resorcinol: Zhuhai Bairui Pharmaceutical Technology Co., Ltd.

[0032] Whitening compound (I): Self-made, preparation method as follows:

[0033] The preparation method of the whitening compound (Ⅰ) includes the following steps:

[0034] S1: Under a nitrogen atmosphere, dissolve 0.01 mol of 3,4-dihydroxymandelic acid in anhydrous DMF (N,N-dimethylformamide), then add 0.012 mol of EDC and 0.012 mol of NHS sequentially, and stir for 0.5 h until activation is complete to form an activated 3,4-dihydroxymandelic acid solution; set aside for later use.

[0035] S2: Under a nitrogen atmosphere, 0.01 mol of 4-aminoresorcinol was dissolved in DMF and slowly added dropwise to the activation solution. The mixture was stirred at room temperature for 2 hours until the reaction was complete. After purification, the whitening compound (Ⅰ) was obtained.

[0036] The purification was performed using high-performance liquid chromatography (HPLC); the mobile phase of the HPLC was water and acetonitrile containing 0.1% trifluoroacetic acid; the chromatographic column was an XBridge BEH C18 OBD™ Prep column. Purification was performed at a flow rate of 4.5 mL / min using a 5 μm, 10 × 250 mm filter. The purified compound was then freeze-dried to obtain whitening compound (Ⅰ) and stored at -20 °C protected from light. The near-infrared spectrum of whitening compound (Ⅰ) is shown in the attached figure in the product manual. Figure 2 As shown.

[0037] Unless otherwise specified, all raw materials used in the embodiments and comparative examples of this invention are commercially available, and the raw materials used in each parallel test are the same.

[0038] Example 1

[0039] A method for preparing a skin whitening and spot-fading skincare product:

[0040] Whitening compound (I), cocoyl glucoside, phospholipids (HSPE), and EDTA were added to deionized water, with the amount of deionized water being twice the amount of the above substances (whitening compound (I), cocoyl glucoside, phospholipids, and EDTA). The mixture was stirred for 5 minutes in a 50°C water bath under nitrogen atmosphere to prepare a mixture. The mixture was then sheared and pulverized at 10,000 rpm for 1 minute under nitrogen atmosphere using a high-speed shear mill. The mixture was then homogenized using a rapid M110-EH homogenizer (Microfluidics, USA) at 8000 bar, repeated 3 times, to obtain a micelle solution. Hyaluronic acid was dissolved in the remaining amount of deionized water to prepare a hyaluronic acid solution. The micelle solution was added to the hyaluronic acid solution and stirred until homogeneous to obtain a whitening and spot-fading skincare product.

[0041] Examples 2-6 were prepared using the same method as Example 1, except that the amounts of each component were adjusted.

[0042] Table 1. Components and parts by mass of Examples 1-6

[0043]

[0044]

[0045] Comparative Examples 1-6

[0046] The preparation method is the same as in Examples 1-6, except that the specific components and their mass fractions are different, as shown in Table 2.

[0047] Table 2. Components and parts by mass of Comparative Examples 1-6

[0048]

[0049] The whitening skincare products or components prepared in Examples 1-6 and Comparative Examples 1-6 were subjected to the following performance tests.

[0050] (1) Skin irritation test:

[0051] Skin irritation tests were conducted on the whitening skincare products used in the examples and comparative examples. The BALB / c-nu test was performed on nude mice, with six mice participating in each sample application. 30 μL of the sample was applied to the back of each mouse (1 × 1 cm) twice daily (9:00 AM and 6:00 PM). The irritation level after application was 0.5 J / cm. 2 Mice were irradiated with ultraviolet light for 10 minutes for 4 consecutive weeks. Their behavior and skin condition were observed daily (compared to the untreated skin area of ​​the same mouse). A blank control group and a negative control group were included (reagent: deionized water). Scoring was performed according to the criteria in the table below:

[0052] Mouse skin condition Rating / points rosy and smooth skin 5 No irritation, normal skin tone 4 dull skin 3 Redness and swelling, mild irritation (scratching by mice) 2 Ulcers, blisters, poor mobility, death 0

[0053] Table 3 Results of skin irritation test 4 weeks later

[0054]

[0055]

[0056] The above mouse experiment results showed that none of the mice in the experimental group experienced adverse reactions, indicating that the whitening skin care product provided by this invention has good biocompatibility, and the whitening compound (I) is non-toxic and non-irritating. In Examples 1-6, the areas where the whitening skin care product was applied to the mice all showed a rosy and smooth appearance, demonstrating that the whitening skin care product has good moisturizing, hydrating, and whitening effects. Comparative Example 1 lacked cocoyl glucoside, Comparative Example 3 lacked menthol, Comparative Example 4 lacked phospholipids, Comparative Example 5 lacked both phospholipids and cocoyl glucoside, and Comparative Example 6 lacked phospholipids, menthol, and cocoyl glucoside, all showing varying degrees of difference in transdermal whitening effects. This is related to the permeation-enhancing effect of menthol and cocoyl glucoside, and the ability of phospholipids and cocoyl glucoside to form cell-membrane-friendly absorption-enhancing micelles. Menthol, cocoyl glucoside, and phospholipids can synergistically promote transdermal absorption.

[0057] Human closed patch test: The test was conducted using human skin patch testing. The specific test method is in accordance with the "Cosmetic Safety Technical Specifications" (2015 edition), as detailed in the table below.

[0058]

[0059] A total of 120 subjects, with an average age of 21 years, were randomly divided into 12 groups. 25 μL of samples from Examples 1-6 and Comparative Examples 1-6 were used in the experiment. The experimental results (see Table 4) showed that samples from Examples 1-6 and Comparative Examples 1-6 all showed negative reactions and did not induce erythema on the skin.

[0060] Table 4. Results of patch test

[0061]

[0062] (2) Transdermal cell absorption test

[0063] Mouse melanoma B16F10 cells were cultured to the logarithmic growth phase using 6-well plates, with 10 cells per well. 5 Cells were seeded, 2 ml of culture medium was added, and the cells were incubated in a CO2 incubator for 24 h. The culture medium was then replaced, and 0.5 ml of each of the samples from Examples 1-6 and Comparative Examples 1-6 was added to each well. Each sample was set up with 6 replicates and incubated for 24 h.

[0064] Cell processing: After trypsin digestion, the cells were pipetted with 2 ml of 50% ethanol aqueous solution and collected into 5 ml EP tubes. The cells were sonicated for 3 min under ice bath conditions, centrifuged at 8000 g, and 0.5 ml of the supernatant was taken. 0.5 ml of methanol was added, and the cells were centrifuged at 10000 g to obtain the supernatant, which is the sample to be tested. The test results are shown in Table 5.

[0065] Detection conditions: High-performance liquid chromatograph: Agilent 1200HPLC, DAD detector; Column: Diol-HILIC column (4.6 mm × 250 mm, 5 μm); Temperature: 30℃; Mobile phase: Mobile phase A: 10 mmol / L ammonium acetate aqueous solution; Mobile phase B: acetonitrile. The ratio of mobile phase A to mobile phase B is 7:3. Flow rate: 1 mL / min, injection volume: 5 μL, and the injection concentration is recorded (see Table 6). The standard curve is shown in Table 5.

[0066] Table 5 Standard Curve

[0067]

[0068] Table 6 Results of Transdermal Cell Absorption Test

[0069]

[0070]

[0071] (3) Inhibition of tyrosinase activity

[0072] The reaction system was prepared as follows: PBS pH 6.8; substrate solutions: 2 mmol / L L-tyrosine solution (PBS pH 6.8) and 4 mmol / L L-dopamine solution (PBS pH 6.8); tyrosinase: 150 U / mL; whitening compound (Ⅰ) was prepared into solutions of different concentrations ranging from 0.01 to 10 μmol / L; phenylethyl resorcinol was prepared into solutions of different concentrations ranging from 0.1 to 50 μmol / L.

[0073] Reaction solution (μL) PBS Substrate liquid Sample to be tested Tyrosinase solution Absorbance value (A) 1 120 40 0 40 <![CDATA[A1]]> 2 160 0 0 40 <![CDATA[A2]]> 3 80 40 40 40 <![CDATA[A3]]> 4 120 0 40 40 <![CDATA[A4]]>

[0074] Add samples to 96-well plates according to the above table, and incubate at 37°C for 5 min. Add 40 μL of tyrosinase to each well at room temperature, and incubate at a constant temperature for 15 min. Record the absorbance values ​​(A1, A2, A3, A4) at 475 nm using a multi-functional microplate reader. Each group is repeated 6 times.

[0075] Inhibition rate = [(A1-A2)-(A3-A4)] / (A1-A2), statistical data (see attached diagram in the instruction manual). Figure 4 (Tyrosinase activity inhibition rate graph), calculate IC50. 50 .

[0076] Whitening compound (Ⅰ) μmol / L Phenethyl resorcinol μmol / L <![CDATA[Tyrosinase monophenolase (IC 50 )]]> 0.12 0.32 <![CDATA[Tyrosinase diphenolase (IC 50 )]]> 9.8 29.7

[0077] (4) DPPH free radical scavenging activity assay

[0078] Whitening compound (I) and phenylethyl resorcinol were dissolved in anhydrous ethanol to prepare test solutions of different concentrations. A DPPH 0.2 mmol / L anhydrous ethanol solution was prepared. A 3 mL assay system was prepared, mixed thoroughly in a cuvette, allowed to stand for 30 min, and the absorbance at 517 nm was measured. Each group was measured three times, and the average value was taken. The clearance rate was calculated as: [1-(A1-A2) / A3]×100%. The IC50 was then calculated. 50 (Table 7)

[0079]

[0080] Table 7 Results of DPPH free radical scavenging effect

[0081]

[0082] (5) Storage stability study

[0083] Examples 1-6 and Comparative Examples 1-6: skin care products were stored in a 25°C constant temperature cabinet away from light. The particle size changes of the samples were measured at 0, 30, and 90 days using a Nano-ZS90 particle size analyzer, and the concentration changes of whitening compound (Ⅰ) in the skin care products were also measured (see the determination of the standard curve above).

[0084] Table 8 Stability Results

[0085]

[0086]

[0087] As shown in Table 8, the particle size and content of whitening compound (Ⅰ) of the highly effective whitening and spot-fading skin care products prepared in Examples 1-6 remained relatively stable after 90 days; the particle size of the highly effective whitening and spot-fading skin care products prepared in Comparative Examples 1 and 4 tended to increase after 90 days, and the dispersion PDI also increased significantly; the content of whitening compound (Ⅰ) decreased in Comparative Example 2.

[0088] The preferred embodiments of the present invention disclosed above are merely illustrative of the invention. These preferred embodiments do not exhaustively describe all details, nor do they limit the invention to the specific implementations described. Clearly, many modifications and variations can be made based on the content of this specification. This specification selects and specifically describes these embodiments to better explain the principles and practical applications of the invention, thereby enabling those skilled in the art to better understand and utilize the invention. The invention is limited only by the claims and their full scope and equivalents.

Claims

1. A skin whitening and spot-fading skincare product, characterized in that, The product comprises, by weight, the following components: 0.1-5 parts of whitening compound (I), 5-10 parts of cocoyl glucoside, 0.5-1 part of menthol, 5-10 parts of phospholipids, 0.5-1 part of EDTA, 0.1-1 part of hyaluronic acid, and 150-200 parts of deionized water. The molecular structure of the whitening compound is as follows: (I): （Ⅰ）。 2. The whitening and spot-fading skincare product as described in claim 1, characterized in that: The preparation method of the whitening compound (Ⅰ) includes the following steps: S1: Under a nitrogen atmosphere, 3,4-dihydroxymandelic acid is dissolved in anhydrous DMF, EDC and NHS are added sequentially, and the mixture is stirred to activate the solution and obtain an activated solution of 3,4-dihydroxymandelic acid. S2: Under a nitrogen atmosphere, the DMF solution of 4-aminoresorcinol was slowly added dropwise to the activation solution, and the reaction was stirred at room temperature until complete. The mixture was then purified by column chromatography to obtain the purified whitening compound (Ⅰ).

3. The whitening and spot-fading skincare product as described in claim 2, characterized in that: The molar ratio of 3,4-dihydroxymandelic acid, EDC and NHS is 1:(1-1.2):(1-1.2).

4. The whitening and spot-fading skincare product as described in claim 2, characterized in that: The molar ratio of 3,4-dihydroxymandelic acid to 4-aminoresorcinol is 1:(1-1.1).

5. The whitening and spot-fading skincare product as described in claim 1, characterized in that: Includes 0.1-3 parts of whitening compound (Ⅰ).

6. The whitening and spot-fading skincare product as described in claim 1, characterized in that, The phospholipid is selected from at least one of 1,2-distearyl-sn-glycerol-3-phosphate ethanolamine, sodium dioleoylphosphatidylserine, egg yolk lecithin, soybean lecithin, distearylphosphatidylcholine, dioleoyl L-α-phosphatidylethanolamine, 1,2-dioleoyl lecithin, and dipalmitoylphosphatidylcholine.

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