Efficient-absorption whitening and spot-fading skin care product and preparation process thereof

Abstract

The invention discloses an efficient-absorption whitening and spot-fading skin care product and a preparation process thereof, and belongs to the technical field of cosmetics, the efficient-absorption whitening and spot-fading skin care product comprises the following components in parts by weight: 0.1-5 parts of a whitening compound (I), 5-10 parts of coco-glucoside, 0.5-1 part of menthol, 5-10 parts of phospholipid, 0.5-1 part of EDTA (Ethylene Diamine Tetraacetic Acid) and 0.1-1 part of hyaluronic acid. The skin-whitening and spot-fading skin-care product provided by the invention has good effects of resisting oxidation and inhibiting tyrosinase activity, and can realize the effects of whitening, fading spots and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and specifically to a skin whitening and freckle-reducing skin care product with high absorption efficiency and its preparation process. Background Art

[0002] The human skin is divided into the epidermis and the 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, and exist in the form of multiple thin lamellar bilayers. Most of the proteins in the stratum corneum are keratin fibers condensed from keratinized cells. Substances with a relative molecular weight greater than 500 are not easily permeable through the stratum corneum.

[0003] Melanocytes are mainly distributed in the epidermis. The raw material required for melanocytes to synthesize melanin is tyrosine. Tyrosine is synthesized into melanin in membrane-bound vesicles under the catalysis of tyrosinase, and melanocytes will transfer it to adjacent keratinocytes. The currently recognized melanin formation pathway is: tyrosine - dopa - dopaquinone - dopachrome - dihydroxyindole - ketole indole melanin. The main current whitening methods are: 1. Directly inhibit the production of melanin. Tyrosinase is the rate-limiting enzyme in the melanin synthesis process, and its activity can be inhibited to achieve a whitening effect. For example, phenethyl resorcinol is commonly used in skin care products of major brands. 2. Antioxidation, by reducing the intermediates in the melanin production process, thereby blocking the production of melanin. 3. Inhibit the transfer of melanin to keratinocytes. 4. Stratum corneum exfoliation to accelerate melanin metabolism. 5. Reduce ultraviolet radiation.

[0004] Although the prior art has clarified the pathway of melanin production and specific whitening methods, the whitening ingredients in current whitening skin care products on the market are still ingredients such as phenethyl resorcinol, arbutin, vitamin C, niacinamide, fruit acids, plant extracts, polypeptides, etc. There has always been a lack of more efficient whitening active ingredients, and no technological breakthroughs in new products have been achieved. Therefore, to meet the market's whitening skin care needs, there is an urgent need to provide a skin whitening and freckle-reducing skin care product that can be highly absorbed. Summary of the Invention

[0005] In order to overcome the deficiencies of the above prior art, the purpose of the present invention is to provide a skin whitening and freckle-reducing skin care product with high absorption efficiency and its preparation process, which has the technical effects of whitening and freckle-reducing, low irritation, skin toning, and moisturizing.

[0006] The present invention is achieved through the following technical scheme: a whitening and spot-lightening skin care product, characterized in that it comprises the following components by weight: 0.1-5 parts of whitening compound (I), 5-10 parts of coconut 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:

[0007]

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

[0009] S1: Under nitrogen atmosphere, 3,4-dihydroxymandelic acid is dissolved in anhydrous DMF (N,N-dimethylformamide) solvent, EDC and NHS are added in sequence, and the mixture is stirred until activation is complete to form a 3,4-dihydroxymandelic acid activated solution for later use.

[0010] S2: In a nitrogen atmosphere, slowly add the DMF solution of 4-aminoresorcinol to the activated solution, stir at room temperature until the reaction is complete, and purify by column chromatography to obtain the purified whitening compound (I).

[0011] The molar ratio of the 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 the 3,4-dihydroxymandelic acid to 4-aminoresorcinol is 1:(1-1.1), preferably 1:1.

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

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

[0015] Beneficial Effects

[0016] The present invention provides a skin care product for whitening and lightening freckles. A new whitening compound (I) is synthesized from 4-aminoresorcinol and 3,4-dihydroxymandelic acid, which has a chemical structure similar to that of phenethyl resorcinol, with a similar double benzene ring and polyphenol hydroxyl structure. Phenethyl resorcinol is a highly effective whitening ingredient and is used in skin care and cosmetics of many brands such as L'Oréal. It has the effects of antioxidation and inhibition of tyrosinase activity. The whitening compound (I) of the present invention has stronger antioxidation activity and tyrosinase inhibition activity performance compared with phenethyl resorcinol, and its water solubility is improved, which is more conducive to its whitening effect in skin cell tissues.

[0017] When cocoyl glucoside is used in combination with phospholipids, stable micelles can be formed to encapsulate the whitening compound (I), and the micelles increase the lipophilicity; cocoyl glucoside and menthol can disrupt the stratum corneum structure and enhance the fluidity of the phospholipid membrane of stratum corneum cells; the synergistic effect of cocoyl glucoside, phospholipids and menthol can greatly improve the transdermal absorption of the whitening ingredient and also has a skin-toning effect. The addition of EDTA can improve the stability of the product and effectively prevent the oxidation and inactivation of the whitening compound (I), which may be related to the ability of EDTA to chelate metal ions.

[0018] The whitening skin care product of the present invention has the effects of highly efficient whitening and lightening freckles, antioxidation and easy transdermal absorption; it uses an aqueous base, has no greasy feeling, and can enhance the moisturizing effect on the skin. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Synthesis route diagram of the whitening compound (I).

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

[0021] Figure 3 Physical picture of the powder of the whitening compound (I).

[0022] Figure 4 Tyrosinase activity inhibition rate diagram. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0023] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention.

[0024] In the embodiments, the test methods used are all conventional methods unless otherwise specified, and the materials, reagents, etc. used can be obtained from commercial channels unless otherwise specified.

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

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

[0027] 3,4-Dihydroxymandelic acid: Purchased from Shanghai Yuanye Bio-Technology Co., Ltd.;

[0028] EDC (1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride): Purchased from Gil 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] Phenylethyl resorcinol: Zhuhai Bairui Pharmaceutical Technology Co., Ltd.

[0032] Whitening compound (I): Self-made, and the preparation method is 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 the solvent anhydrous DMF (N,N-Dimethylformamide), and sequentially add 0.012 mol of EDC and 0.012 mol of NHS, and stir for 0.5 h until the activation is completed to form a 3,4-dihydroxymandelic acid activation solution; set aside.

[0035] S2: Under a nitrogen atmosphere, dissolve 0.01 mol of 4-aminoresorcinol in DMF, and slowly drop it into the activation solution, and stir at room temperature for 2 h until the reaction is complete, and purify to obtain the whitening compound (I).

[0036] The purification is carried out by high performance liquid chromatography; the mobile phase of the high performance liquid chromatography is water and acetonitrile containing 0.1% trifluoroacetic acid; the chromatographic column is XBridge BEH C18 OBDTM Prep column, 5 μm, 10×250 mm, and purified at a flow rate of 4.5 mL / min. After purification, it is freeze-dried to obtain the whitening compound (I), and stored at -20°C in the dark. The near-infrared spectrum of the whitening compound (I) is as shown in the accompanying drawings of the specification Figure 2 as shown.

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

[0038] Example 1

[0039] Preparation method of a skin care product for whitening and freckle reduction:

[0040] Add a whitening compound (I), cocoyl glucoside, phospholipid (HSPE), and EDTA to deionized water. The amount of deionized water used is twice the amount of the above substances (whitening compound (I), cocoyl glucoside, phospholipid, EDTA). Under a nitrogen atmosphere, water bath at 50 °C and stir for 5 min to prepare a mixed solution; under a nitrogen atmosphere, shear and crush with a high-speed shearing machine at 10,000 rpm for 1 min; quickly homogenize with a M110-EH type homogenizer (Microfluidics, USA) under the condition of 8000 bar for 3 cycles to obtain a micelle solution. Dissolve hyaluronic acid in the remaining amount of deionized water in the prescription to obtain a hyaluronic acid solution. Add the above micelle solution to the hyaluronic acid solution and stir evenly to obtain a skin care product for whitening and freckle reduction.

[0041] The preparation methods of Examples 2 - 6 are the same as that of Example 1, except that the amounts of each component are adjusted.

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

[0043]

[0044]

[0045] Comparative Examples 1 - 6

[0046] The preparation methods are the same as those of Examples 1 - 6, except that the specific components and their parts by mass formulations are different, as shown in Table 2 specifically.

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

[0048]

[0049] Conduct the following performance tests on the whitening skin care products or components in the products prepared in Examples 1 - 6 and Comparative Examples 1 - 6.

[0050] (1) Skin irritation test:

[0051] Conduct a skin irritation test on the whitening skin care products of the examples and comparative examples. Use the nude mouse BALB / c-nu test. For each sample, 6 mice are in parallel. Apply the sample (30 μL) to the back of the mouse (1×1 cm), once in the morning and once in the evening (9:00 / 18:00); irradiate with ultraviolet light of 0.5 J / cm 2 for 10 min after application, continuously for 4 weeks, and observe the behavior and skin condition of the mice every day (compared with the skin of the non-applied area of the same mouse); set up a blank group and a negative control group (the reagent is: deionized water). Score according to the following table standard:

[0052] Mouse skin condition Score / Points Skin is rosy and smooth 5 No irritation, normal skin color 4 Skin is dull 3 Red rash and edema, slight irritation (mouse scratching) 2 Ulcer, blister, poor activity, death 0

[0053] Table 3 Results after 4 - week skin irritation test

[0054]

[0055]

[0056] The above - mentioned mouse test results showed that no adverse reactions occurred in the test - group mice, indicating that the whitening skin - care product provided by the present invention has good biological safety, and the whitening compound (I) is non - toxic and non - irritating. For the mice in Examples 1 - 6, the parts of the mice smeared with the whitening skin - care product were all shown to be ruddy and smooth, demonstrating that the whitening skin - care product has good effects of moisturizing the skin, retaining moisture and whitening. In Comparative Example 1, due to the lack of cocoyl glucoside, Comparative Example 3 lacked menthol, Comparative Example 4 lacked phospholipids, Comparative Example 5 lacked phospholipids and cocoyl glucoside, and Comparative Example 6 lacked phospholipids, menthol and cocoyl glucoside, and different degrees of differences in transdermal whitening effects occurred. This is related to the penetration - promoting effects of menthol and cocoyl glucoside, and the fact that phospholipids and cocoyl glucoside can form cell - membrane - friendly absorption - promoting micelles. Menthol, cocoyl glucoside and phospholipids can synergistically promote transdermal absorption.

[0057] Human occlusive patch test: The human skin patch test was used for testing. The specific test method refers to the "Technical Specifications for Cosmetics Safety" (2015 Edition), as shown in the following table.

[0058]

[0059] A total of 120 subjects, with an average age of 21 years old, were randomly divided into 12 groups. Samples of Examples 1 - 6 and Comparative Examples 1 - 6, each with 25 μL, were used for the test. The test results recorded (see Table 4) showed that the samples of Examples 1 - 6 and Comparative Examples 1 - 6 all showed negative reactions and did not cause skin erythema reactions.

[0060] Table 4 Patch test results

[0061]

[0062] (2) Transdermal cell absorption test

[0063] Mouse melanoma B16F10 cells were cultured to the logarithmic growth phase. A 6 - well culture plate was used, and 10 5 cells were inoculated in each well, with 2 ml of culture medium. They were cultured in a carbon dioxide incubator for 24 h, the culture medium was changed, and 0.5 ml of the samples of Examples 1 - 6 and Comparative Examples 1 - 6 were added to each well. Six replicates were set for each sample, and they were cultured for 24 h.

[0064] Cell treatment: After trypsin digestion, the cells were pipetted with 2 ml of 50% ethanol aqueous solution, and the cells were collected into a 5 ml EP tube. Ultrasonic fragmentation was performed for 3 min under ice bath conditions, centrifuged at 8000 g, 0.5 ml of the supernatant was taken, 0.5 ml of methanol was added, and the supernatant was obtained by centrifugation at 10000 g, which was the sample to be detected. The test results are shown in Table 5.

[0065] Detection conditions: High performance liquid chromatograph: Agilent 1200 HPLC, DAD detector; chromatographic column: Diol-HILIC column (4.6 mm × 250 mm, 5 μm); temperature: 30 °C; mobile phase; mobile phase A: 10 mmol / L ammonium acetate aqueous solution; mobile phase B: acetonitrile. Mobile phase A: Mobile phase B was 7:3. The flow rate was 1 ml / min, the injection volume was 5 μl, and the injection concentration was recorded, as shown in 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) Tyrosinase activity inhibition

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

[0073] Reaction solution (μL) PBS Substrate solution 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 a 96-well plate according to the above table, incubate in a water bath at 37 °C for 5 min; add 40 μL of tyrosinase to each well at room temperature, after incubating in a constant temperature water bath for 15 min, the absorbance values (A1, A2, A3, A4) at 475 nm were recorded with a multi-functional microplate reader, and each group was parallel 6 times.

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

[0076] Whitening compound (Ⅰ) μmol / L Phenylethyl 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] Dissolve whitening compound (I) and phenethyl resorcinol in anhydrous ethanol to prepare test solutions of different concentrations. Prepare an anhydrous ethanol solution of 0.2mmol / L DPPH. Prepare 3mL of the activity test system, mix well in a cuvette, let stand for 30min, measure the absorbance at 517nm, measure 3 times for each group, take the average value, and calculate: Clearance = [1-(A1-A2) / A3] × 100%, calculate IC 50 (Table 7).

[0079]

[0080] Table 7 DPPH free radical scavenging effect results

[0081]

[0082] (5) Storage stability study

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

[0084] Table 8 Stability results

[0085]

[0086]

[0087] It can be seen from Table 8 that after 90 days, the particle size and the content of whitening compound (I) of the high-efficiency whitening and freckle-removing skin care products prepared in Examples 1-6 remain relatively stable; after 90 days, the particle size of the high-efficiency whitening and freckle-removing skin care products prepared in Comparative Examples 1 and 4 tend to become larger, and the dispersion degree PDI also increases significantly; the content of whitening compound (I) in Comparative Example 2 is reduced.

[0088] The preferred embodiments of the present invention disclosed above are only used to help illustrate the present invention. The preferred embodiments do not describe all the details in detail, nor do they limit the invention to the specific implementation methods described. Obviously, many modifications and changes can be made according to the content of this specification. This specification selects and specifically describes these embodiments in order to better explain the principles and practical applications of the present invention, so that those skilled in the art can understand and use the present invention well. The present invention is limited only by the claims and their full scope and equivalents.

Claims

1. A skin care product for whitening and fading spots, characterized in that, By weight, it includes 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 phospholipid, 0.5 - 1 part of EDTA, 0.1 - 1 part of hyaluronic acid, 150 - 200 parts of deionized water. The molecular structure of the whitening compound is as follows formula (Ⅰ):

2. The whitening and spot-lightening skin care product according to claim 1, characterized in that: The preparation method of the whitening compound (Ⅰ) includes the following steps: S1: Under a nitrogen atmosphere, dissolve 3,4 - dihydroxy mandelic acid in the solvent anhydrous DMF (N,N - dimethylformamide), and successively add EDC and NHS, stir for activation to obtain a 3,4 - dihydroxy mandelic acid activation solution; S2: Slowly drop the DMF solution of 4 - aminoresorcinol into the activation solution under a nitrogen atmosphere, stir at room temperature until the reaction is complete, and purify by column chromatography to obtain the purified whitening compound (Ⅰ).

3. The skin-whitening and freckle-reducing skin care product according to claim 2, characterized in that: The molar dosage ratio of 3,4 - dihydroxy mandelic acid, EDC and NHS is 1:(1 - 1.2):(1 - 1.2).

4. The whitening and spot-lightening skin care product according to claim 2, characterized in that: The molar dosage ratio of 3,4 - dihydroxy mandelic acid to 4 - aminoresorcinol is 1:(1 - 1.1).

5. The skin whitening and freckle lightening skin care product according to claim 1, characterized in that: 0.1 - 3 parts of whitening compound (Ⅰ).

6. The skin-whitening and freckle-reducing skin care product according to claim 1, wherein The phospholipid is selected from at least one of 1,2 - distearoyl - sn - glycero - 3 - phosphoethanolamine, sodium dioleoyl phosphatidylserine, egg yolk lecithin, soybean lecithin, distearoyl phosphatidylcholine, dioleoyl L - α - phosphatidylethanolamine, 1,2 - dioleoyl lecithin, dipalmitoyl phosphatidylcholine.

7. The preparation method of the skin whitening and freckle lightening skin care product according to any one of claims 1-6, characterized in that: Add the whitening compound (Ⅰ), cocoyl glucoside, phospholipid, and EDTA into deionized water. The dosage of pure water is twice the total dosage of the whitening compound (Ⅰ), cocoyl glucoside, phospholipid, and EDTA. Under a nitrogen atmosphere, perform water bath, high - speed shearing, and homogenize with a high - pressure homogenizer to prepare a micellar solution; Dissolve hyaluronic acid in the remaining deionized water to prepare a hyaluronic acid solution; Add the above - mentioned micellar solution into the hyaluronic acid solution and stir evenly.

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