Whitening nanocomposition, preparation method therefor, and use thereof

Abstract

A whitening nanocomposition, a preparation method therefor, and use thereof. The whitening nanocomposition comprises an active ingredient and a nanocarrier. The active ingredient is coated and / or adsorbed by the nanocarrier. The active ingredient comprises the following components: 1-15 parts by weight of α-arbutin, 1-15 parts by weight of nicotinamide, 1-10 parts by weight of 3-o-ethyl ascorbic acid, 0.1-1 part by weight of a Centalla asiatica extract, and 1-5 parts by weight of retinyl palmitate. The nanocarrier comprises the following components: 5-10 parts by weight of an emulsifier, 5-20 parts by weight of a polyol, 0.1-5 parts by weight of a phospholipid, and 40-60 parts by weight of water. The whitening nanocomposition has good skin permeability and retention performance and enables high-concentration enrichment, prolonged retention, and sustained release or controlled release within a target tissue, so as to be effectively taken in by target cells, thereby improving bioavailability and enhancing the whitening effect.

Description

A whitening nano composition, a preparation method and application thereof TECHNICAL FIELD

[0001] The present application relates to the field of cosmetic or similar toiletry formulation technology, in particular to a whitening nano composition, a preparation method and application thereof. BACKGROUND

[0002] The pigments in human skin mainly include black melanin, red hemoglobin and yellow carotene, among which melanin is the key factor affecting skin color. External factors such as ultraviolet rays and environmental pollution and internal factors such as hormones and inflammation levels jointly act on the skin, affecting the secretion of melanin by melanocytes through different mechanisms and pathways. The current whitening ingredients mainly reduce the amount of melanin in the skin through different pathways to achieve the effect of whitening, and the specific pathways mainly include: inhibiting the activity of tyrosinase, accelerating the metabolism of keratinocytes with pigmentation, reducing the stimulation of ultraviolet rays or free radicals on melanocytes, etc.

[0003] However, there are various whitening products on the market, and most of them are difficult to achieve satisfactory whitening effect for consumers, the main reasons of which include: (1) the existing whitening products have a single mechanism of action and certain irritability, for example, some products have strong inhibitory effect on tyrosinase activity and keratolysis, but lack of active ingredients for repairing the skin, resulting in damage to the skin barrier; (2) the stability of whitening functional ingredients is poor, which is easy to be oxidized and inactivated when directly applied to cosmetic products, thereby making it difficult to play a whitening role; (3) due to the barrier function of the skin itself, it is difficult for whitening functional ingredients to penetrate the stratum corneum into the dermis, and it is even more difficult to accumulate in the target site of the skin to reach an effective concentration range, thereby resulting in poor actual whitening effect. SUMMARY

[0004] The present application aims to overcome the shortcomings of the prior art and provide a whitening nano composition, a preparation method and application thereof.

[0005] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:

[0006] In a first aspect, the present application provides a whitening nano composition, which comprises active ingredients and a nano carrier, wherein the active ingredients are coated and / or adsorbed by the nano carrier.

[0007] The active ingredients comprise the following components by weight fraction: 1-15 parts of alpha-arbutin, 1-15 parts of nicotinamide, 1-10 parts of 3-o-ethyl ascorbic acid, 0.1-1 parts of asiaticoside extract, and 1-5 parts of retinol palmitate.

[0008] The nanocarrier comprises the following components in parts by weight: 5-10 parts of emulsifier, 5-20 parts of polyol, 0.1-5 parts of phospholipid, and 40-60 parts of water.

[0009] The whitening nanocomposition of the present application takes alpha-arbutin, nicotinamide, 3-o-ethyl ascorbic acid, gynura extract and retinol palmitate as the main whitening active ingredients, and combines the above-mentioned whitening active ingredients with the nanocarrier of specific components. This not only improves the water dispersibility of the whitening nanocomposition to effectively improve the solubility of the whitening active ingredients, but also enhances the stability of the whitening nanocomposition to avoid unnecessary degradation or inactivation of the whitening active ingredients before storage or use, thereby improving the concentration of the whitening active ingredients in the whitening product. At the same time, the combination of the above-mentioned whitening active ingredients with the nanocarrier of specific components can also reduce the irritation of the whitening active ingredients, so that they can reach a sufficient concentration in the product to exert the corresponding functional effects. Moreover, under the action of the nanocarrier, the whitening active ingredients in the whitening nanocomposition can efficiently penetrate the skin barrier, rapidly reach the basal layer where melanocytes are distributed and the dermis layer located deep in the skin, and be concentrated in the target tissue for a long time, thus being effectively taken up by target cells, improving the bioavailability of the whitening active ingredients, prolonging the action time, and thereby enhancing the whitening effect.

[0010] The above-mentioned alpha-arbutin, nicotinamide, 3-o-ethyl ascorbic acid, gynura extract and retinol palmitate have a synergistic effect on multiple effects and multiple targets. Among them, alpha-arbutin contains a strong hydrophilic glucose residue, which can effectively retain water in the skin and easily penetrate the outer layer of the skin. At the same time, the phenolic group contained in alpha-arbutin can effectively inhibit the biological activity of tyrosinase to reduce the formation of cellular melanin. Nicotinamide not only can inhibit the formation of melanin particles, but also can effectively inhibit the transfer of melanin to keratinocytes. When part of the melanin inevitably reaches the surface of the skin, nicotinamide can promote the shedding of cells containing melanin by accelerating the renewal rate of skin cells. At the same time, nicotinamide can also effectively inhibit the activity of tyrosinase, prevent the transfer of melanin to the stratum corneum, and promote the metabolism of cells containing melanin. In addition, nicotinamide also has antioxidant properties, which can reduce the content of reactive oxygen species (ROS) in the skin, thereby significantly reducing pigmentation, brightening the skin, and improving the skin whitening degree.

[0011] 3-o-ethyl ascorbic acid can protect the skin from damage by free radicals to slow down the aging process of the skin, and can also inhibit the activity of tyrosinase to reduce the formation of melanin, and can inhibit the formation of DHI (dihydroxyindole)-melanin and DHICA (dihydroxyindole acetic acid)-melanin from tyrosine.

[0012] The extract of Centella asiatica contains rich polysaccharides, vitamins, flavonoids and other substances. These substances can help the human body to remove free radicals, reduce skin pigmentation, and have antibacterial, anti-inflammatory, and promote fibroblast regeneration and other pharmacological effects. In addition, these substances not only have a certain promoting effect on human fibroblast proliferation and collagen synthesis to improve scars, but also can inhibit oxidative stress, thereby playing an anti-inflammatory repair role.

[0013] The main components of the above-mentioned Centella asiatica extract are pentacyclic triterpenoids, including asiaticoside, hydroxyl asiaticoside and its aglycone, asiatic acid and hydroxyl asiatic acid, which can be purchased or prepared by existing preparation methods.

[0014] Retinol palmitate has antioxidant function, can accelerate the renewal speed of skin deep layer cells, also can promote the metabolism of growth factors, thereby improving skin metabolism, promoting skin to produce more epidermal proteins, making skin more elastic. At the same time, retinol palmitate not only can make the keratinocytes that have formed fall off with melanin as soon as possible, but also has the ability to inhibit the formation of melanin, thereby having good whitening effect.

[0015] Optionally, in the above-mentioned active ingredients, the α-arbutin can be 2 parts, 4 parts, 6 parts, 8 parts, 10 parts, 12 parts, 14 parts; the nicotinamide can be 2 parts, 4 parts, 6 parts, 8 parts, 10 parts, 12 parts, 14 parts; the 3-o-ethyl ascorbic acid can be 3 parts, 5 parts, 7 parts, 9 parts; the Centella asiatica extract can be 0.3 parts, 0.5 parts, 0.7 parts, 0.9 parts; the retinol palmitate can be 1.5 parts, 2.5 parts, 3.5 parts, 4.5 parts.

[0016] Optionally, in the above-mentioned nano carrier, the emulsifier can be 5.5 parts, 6.5 parts, 7.5 parts, 8.5 parts, 9.5 parts; the polyol can be 7 parts, 9 parts, 11 parts, 13 parts, 15 parts, 17 parts, 19 parts; the phospholipid can be 0.5 parts, 1 parts, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts; the water can be 43 parts, 45 parts, 47 parts, 49 parts, 51 parts, 53 parts, 55 parts, 57 parts, 59 parts.

[0017] The average particle size of the above-mentioned whitening nano composition is 20-40 nm, which can be measured by a particle size instrument.

[0018] As a preferred embodiment of the whitening nano composition of the present application, the α-arbutin is 8-10 parts, the nicotinamide is 5-10 parts, the 3-o-ethyl ascorbic acid is 3-5 parts, the Centella asiatica extract is 0.1-1 parts, and the retinol palmitate is 1-3 parts by weight.

[0019] As a preferred embodiment of the whitening nano-composition of the present application, the emulsifier is 5-10 parts, the polyol is 9-18 parts, the phospholipid is 0.5-1 part, and the water is 46-55 parts by weight.

[0020] As a preferred embodiment of the whitening nano-composition of the present application, the mass ratio of the active ingredient to the nano-carrier is (15-30):(65-80), preferably (19.1-28.5):(69-78.5).

[0021] Alternatively, the active ingredient can be 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts by weight; and the nano-carrier can be 70 parts, 72 parts, 74 parts, 76 parts, 78 parts.

[0022] As a preferred embodiment of the whitening nano-composition of the present application, the emulsifier includes at least one of polyoxyethylene castor oil emulsifier, polyoxyethylene hydrogenated castor oil emulsifier, polyglycerol emulsifier, poloxamer, and coco-glucoside.

[0023] Alternatively, the polyoxyethylene castor oil emulsifier can be at least one of polysorbate-20, polysorbate-60, and polysorbate-80, the polyoxyethylene hydrogenated castor oil emulsifier can be at least one of PEG-40 hydrogenated castor oil and PEG-60 hydrogenated castor oil, and the polyglycerol emulsifier can be at least one of polyglyceryl fatty acid ester and polyglyceryl-10 stearate.

[0024] As a preferred embodiment of the whitening nano-composition of the present application, the polyol includes at least one of glycerol, propylene glycol, butylene glycol, 1,3-propanediol, 1,2-pentanediol, 1,2-hexanediol, dipropylene glycol, isopropyl alcohol, polyethylene glycol-200, PPG-10 sorbitol, and octyldodecanol.

[0025] As a preferred embodiment of the whitening nano-composition of the present application, the phospholipid includes at least one of hydrogenated lecithin, lecithin, and soybean lecithin.

[0026] In a second aspect, the present application provides a preparation method of the above-mentioned whitening nano-composition, which includes the following steps:

[0027] S1, uniformly mixing α-arbutin, nicotinamide, 3-o-ethyl ascorbic acid, and asiaticoside extract with phospholipid, water, and a part of polyol to obtain a mixed solution I;

[0028] S2, uniformly mixing retinol palmitate with emulsifier and the remaining polyol to obtain a mixed solution II;

[0029] S3, homogenizing the mixture I in S1 and the mixture II in S2, to obtain the whitening nano-composition.

[0030] The homogenization in step S3 can be performed by a high-pressure homogenizer, and the pressure of the high-pressure homogenizer can be 800-1000 bar.

[0031] As a preferred embodiment of the preparation method of the whitening nano-composition, step S1 is specifically: mixing a- arbutin, nicotinamide, 3-o-ethyl ascorbic acid and asiaticoside extract, phospholipid, water and part of polyol, stirring at 40-65℃ and 150-200rpm for 10-30min, to obtain the mixture I.

[0032] As a preferred embodiment of the preparation method of the whitening nano-composition, step S2 is specifically: mixing retinol palmitate, emulsifier and the remaining polyol, stirring at 40-65℃ and 150-200rpm for 10-30min, to obtain the mixture II.

[0033] As a preferred embodiment of the preparation method of the whitening nano-composition, step S3 is specifically: mixing the mixture I in S1 and the mixture II in S2, stirring at 40-65℃ and 200-270rpm for 10-30min, homogenizing, to obtain the whitening nano-composition.

[0034] In a third aspect, the present application provides the use of the above-mentioned whitening nano-composition in the preparation of skin care products or cosmetics.

[0035] In a fourth aspect, the present application provides a whitening skin care product or whitening cosmetic comprising the above-mentioned whitening nano-composition.

[0036] As a preferred embodiment of the whitening skin care product or whitening cosmetic, the mass fraction of the whitening nano-composition in the whitening skin care product or whitening cosmetic is 0.1%-30%.

[0037] Optionally, the mass fraction of the whitening nano-composition in the whitening skin care product or whitening cosmetic can be 0.5%, 5%, 10%, 15%, 20%, 25%; and the form of the whitening skin care product or whitening cosmetic includes but is not limited to toner, cream, emulsion, essence and gel.

[0038] Compared with the prior art, the present application has the following beneficial effects:

[0039] The whitening nano composition of the present application takes alpha-arbutin, nicotinamide, 3-o-ethyl ascorbic acid, asiaticoside extract and retinol palmitate as the main whitening active ingredients, and combines the above whitening active ingredients with the nano carrier of specific components, which can not only effectively improve the solubility of the whitening active ingredients, but also enhance the stability of the whitening nano composition, and at the same time, can reduce the irritation of the whitening active ingredients so that they can reach a sufficient concentration in the product to play the corresponding functional effect. Moreover, the whitening active ingredients in the whitening nano composition can efficiently penetrate the skin barrier, quickly reach the basal layer where melanocytes are distributed and the dermis layer located in the deep part of the skin, and be concentrated in the target tissue for a long time, slow release, effectively taken up by target cells, improve the bioavailability of the whitening active ingredients, prolong the action time, and thus enhance the whitening effect. BRIEF DESCRIPTION OF DRAWINGS

[0040] Figure 1 is a chart of the chicken embryo allantoic membrane irritation test of the whitening nano composition in Examples 1-5 respectively diluted 10 times with water;

[0041] Figure 2 is a chart of the skin cumulative permeation amount of the whitening nano composition in Example 1 and the whitening free composition in Comparative Example 8;

[0042] Figure 3 is a chart of the skin storage amount of the whitening nano composition in Example 1 and the whitening free composition in Comparative Example 8 (compared with the free composition, “**” indicates P<0.01);

[0043] Figure 4 is a chart of the fluorescence penetration depth of free RhoB and RhoB nano carrier corresponding to the whitening free composition in Comparative Example 8 and the whitening nano composition in Example 1 at different times;

[0044] Figure 5 is a chart of the relative fluorescence intensity of free RhoB and RhoB nano carrier corresponding to the whitening free composition in Comparative Example 8 and the whitening nano composition in Example 1 at different times;

[0045] Figure 6 is a chart of the relative tyrosinase activity of B16F10 cells corresponding to the samples of the blank control group, the model group and the experimental group of the present application (compared with the model group, “*” indicates P<0.05, “**” indicates P<0.01; compared with Comparative Example 6, “&&” indicates P<0.01; compared with Comparative Example 8, “##” indicates P<0.01);

[0046] Figure 7 is a chart of the intracellular relative melanin content of B16F10 cells corresponding to the samples of the blank control group, the model group and the experimental group of the present application (compared with the model group, “*” indicates P<0.05, “**” indicates P<0.01; compared with Comparative Example 6, “&&” indicates P<0.01; compared with Comparative Example 8, “##” indicates P<0.01);

[0047] Figure 8 is a face skin color contrast chart and a face pigment and spot contrast chart of volunteer 1 using the sample of the experimental group on the 0th day, the 14th day and the 28th day;

[0048] Figure 9 is a face skin color contrast chart and a face pigment and spot contrast chart of volunteer 2 using the sample of the experimental group on the 0th day, the 14th day and the 28th day. DETAILED DESCRIPTION

[0049] For better illustrating the purpose, technical scheme and advantages of the present application, the present application will be further described in combination with specific examples.

[0050] Other materials, reagents and the like used in the examples can be obtained from commercial channels if not specifically stated.

[0051] Example 1

[0052] In one embodiment of the whitening nano-composition of the present application, the whitening nano-composition described in the present embodiment comprises active ingredients and nano-carriers, and the active ingredients are coated and / or adsorbed by the nano-carriers.

[0053] The active ingredients comprise the following components by weight fraction:

[0054] 10 parts of α-arbutin, 10 parts of nicotinamide, 5 parts of 3-o-ethyl ascorbic acid, 0.5 parts of centella asiatica extract, and 3 parts of retinol palmitate;

[0055] The nano-carriers comprise the following components by weight fraction:

[0056] 10 parts of emulsifier (PEG-40 hydrogenated castor oil), 15 parts of polyol (10 parts of glycerol + 5 parts of octyldodecanol), 0.5 parts of phospholipid (lecithin), and 46 parts of water.

[0057] The preparation method of the whitening nano-composition described in the present embodiment is as follows:

[0058] S1, 10 parts of α-arbutin, 10 parts of nicotinamide, 5 parts of 3-o-ethyl ascorbic acid, 0.5 parts of centella asiatica extract, 0.5 parts of lecithin, 10 parts of glycerol and 46 parts of water are stirred at 45℃ and 150-200rpm for 15min to form a uniform and clear liquid A;

[0059] S2, 10 parts of PEG-40 hydrogenated castor oil, 5 parts of octyldodecanol and 3 parts of retinol palmitate are stirred at 45℃ and 150-200rpm for 15min to form a uniform and clear liquid B;

[0060] S3, liquid A is added to liquid B, and stirred at 200-270 rpm for 15 min to form a uniform liquid C, and then liquid C is homogenized twice at 800 bar in a high-pressure homogenizer to obtain the whitening nano composition.

[0061] Example 2

[0062] In one embodiment of the whitening nano composition, the whitening nano composition comprises active ingredients and a nano carrier, and the active ingredients are coated and / or adsorbed by the nano carrier.

[0063] The active ingredients, by weight fraction, comprise the following components:

[0064] Alpha-arbutin 8 parts, nicotinamide 6 parts, 3-o-ethyl ascorbic acid 4 parts, ginseng extract 0.1 part, retinol palmitate 1 part;

[0065] The nano carrier, by weight fraction, comprises the following components:

[0066] Emulsifier (PEG-40 hydrogenated castor oil) 5 parts, polyol 18 parts (15 parts glycerol + 3 parts octyldodecanol), phospholipid (lecithin) 0.5 parts, water 55 parts.

[0067] The preparation method of the whitening nano composition in this embodiment is the same as that in Example 1, only the amount of each component is different.

[0068] Example 3

[0069] In one embodiment of the whitening nano composition, the whitening nano composition comprises active ingredients and a nano carrier, and the active ingredients are coated and / or adsorbed by the nano carrier.

[0070] The active ingredients, by weight fraction, comprise the following components:

[0071] Alpha-arbutin 8 parts, nicotinamide 8 parts, 3-o-ethyl ascorbic acid 3 parts, ginseng extract 1 part, retinol palmitate 1 part;

[0072] The nano carrier, by weight fraction, comprises the following components:

[0073] Emulsifier (PEG-40 hydrogenated castor oil) 6 parts, polyol 9 parts (8 parts glycerol + 1 part octyldodecanol), phospholipid (lecithin) 1 part, water 53 parts.

[0074] The preparation method of the whitening nano composition in this embodiment is the same as that in Example 1, only the amount of each component is different.

[0075] Example 4

[0076] An embodiment of the whitening nano-composition of the present application, the whitening nano-composition of the present embodiment comprises active ingredients and nano-carriers, the active ingredients are coated and / or adsorbed by the nano-carriers;

[0077] The active ingredients, by weight fraction, comprise the following components:

[0078] Alpha-arbutin 8 parts, nicotinamide 5 parts, 3-o-ethyl ascorbic acid 3 parts, ginkgo biloba extract 0.5 parts, retinol palmitate 3 parts;

[0079] The nano-carriers, by weight fraction, comprise the following components:

[0080] Emulsifier (PEG-40 hydrogenated castor oil) 8 parts, polyol 13 parts (8 parts of glycerol + 5 parts of octyldodecanol), phospholipid (lecithin) 0.5 parts, water 55 parts.

[0081] The preparation method of the whitening nano-composition of the present embodiment is the same as that of embodiment 1, only the amount of each component is different.

[0082] Embodiment 5

[0083] An embodiment of the whitening nano-composition of the present application, the whitening nano-composition of the present embodiment comprises active ingredients and nano-carriers, the active ingredients are coated and / or adsorbed by the nano-carriers;

[0084] The active ingredients, by weight fraction, comprise the following components:

[0085] Alpha-arbutin 10 parts, nicotinamide 8 parts, 3-o-ethyl ascorbic acid 5 parts, ginkgo biloba extract 0.5 parts, retinol palmitate 3 parts;

[0086] The nano-carriers, by weight fraction, comprise the following components:

[0087] Emulsifier (PEG-40 hydrogenated castor oil) 10 parts, polyol 20 parts (15 parts of glycerol + 5 parts of octyldodecanol), phospholipid (lecithin) 0.5 parts, water 50 parts.

[0088] The preparation method of the whitening nano-composition of the present embodiment is the same as that of embodiment 1, only the amount of each component is different.

[0089] Embodiment 6

[0090] An embodiment of the whitening nano-composition of the present application, the whitening nano-composition of the present embodiment is basically the same as that of embodiment 1, the difference lies in that the emulsifier in the nano-carrier is polysorbate-80, the polyol is glycerol, and the phospholipid is soy lecithin.

[0091] The preparation method of the whitening nano-composition of the present example is the same as that of Example 1, only the nano-carrier is different.

[0092] Comparative Example 1

[0093] The whitening nano-composition of the present example is a comparative example of the whitening nano-composition of the present application. The whitening nano-composition of the present comparative example is basically the same as that of Example 1, only the active ingredient is different. The active ingredient is α-arbutin (28.5 parts).

[0094] The preparation method of the whitening nano-composition of the present comparative example is the same as that of Example 1, only the active ingredient is different.

[0095] Comparative Example 2

[0096] The whitening nano-composition of the present example is a comparative example of the whitening nano-composition of the present application. The whitening nano-composition of the present comparative example is basically the same as that of Example 1, only the active ingredient is different. The active ingredient is nicotinamide (28.5 parts).

[0097] The preparation method of the whitening nano-composition of the present comparative example is the same as that of Example 1, only the active ingredient is different.

[0098] Comparative Example 3

[0099] The whitening nano-composition of the present example is a comparative example of the whitening nano-composition of the present application. The whitening nano-composition of the present comparative example is basically the same as that of Example 1, only the active ingredient is different. The active ingredient is 3-o-ethyl ascorbic acid (28.5 parts).

[0100] The preparation method of the whitening nano-composition of the present comparative example is the same as that of Example 1, only the active ingredient is different.

[0101] Comparative Example 4

[0102] The whitening nano-composition of the present example is a comparative example of the whitening nano-composition of the present application. The whitening nano-composition of the present comparative example is basically the same as that of Example 1, only the active ingredient is different. The active ingredient is Centella asiatica extract (28.5 parts).

[0103] The preparation method of the whitening nano-composition of the present comparative example is the same as that of Example 1, only the active ingredient is different.

[0104] Comparative Example 5

[0105] The whitening nano-composition of the present example is a comparative example of the whitening nano-composition of the present application. The whitening nano-composition of the present comparative example is basically the same as that of Example 1, only the active ingredient is different. The active ingredient is retinol palmitate (28.5 parts).

[0106] The preparation method of the whitening nano-composition of the present comparative example is the same as that of Example 1, only the active ingredient is different.

[0107] Comparative Example 6

[0108] A comparative example of the whitening nano-composition of the present application, the whitening nano-composition of the comparative example is basically the same as that of Example 1, except that the active ingredients include the following components by weight fraction:

[0109] 10 parts of α-arbutin, 10 parts of nicotinamide, 5 parts of 3-o-ethyl ascorbic acid, and 0.5 parts of Gynostemma pentaphyllum extract;

[0110] The preparation method of the whitening nano-composition of the comparative example is the same as that of Example 1, except that the active ingredients are different.

[0111] Comparative Example 7

[0112] A comparative example of the whitening nano-composition of the present application, the whitening nano-composition of the comparative example is basically the same as that of Example 1, except that the active ingredients include the following components by weight fraction:

[0113] The preparation method of the whitening nano-composition of the comparative example is the same as that of Example 1, except that the active ingredients are different.

[0114] Comparative Example 8

[0115] A comparative example of the whitening free composition of the present application, the whitening free composition of the comparative example includes the following components by weight fraction:

[0116] 10 parts of α-arbutin, 10 parts of nicotinamide, 5 parts of 3-o-ethyl ascorbic acid, 0.5 parts of Gynostemma pentaphyllum extract, 3 parts of retinol palmitate, and 71.5 parts of solvent (consisting of dimethyl sulfoxide and water in a mass ratio of 1:9).

[0117] The preparation method of the whitening free composition of the comparative example is:

[0118] Stir α-arbutin, nicotinamide, 3-o-ethyl ascorbic acid, Gynostemma pentaphyllum extract, retinol palmitate, and solvent at 45°C and 150-200 rpm for 15 min to obtain the whitening free composition.

[0119] Table 1 Weight fraction of each component in the whitening nano-composition of Examples 1-6

[0120] Note: The average particle size in the above Table 1 refers to the average value obtained after testing the sample 3 times using a particle size instrument.

[0121] Performance test

[0122] 1. Stability test

[0123] The whitening nano-compositions in Examples 1-6 were randomly divided into groups A, B, C and D, respectively, and group A was placed at -20℃ for 3 months, group B was placed at room temperature for 3 months, group C was placed at 4℃ for 3 months, and group D was placed at 45℃ for 3 months, and then whether delamination or chromatography occurred was observed. The experimental results showed that the whitening nano-compositions in each example did not appear delamination or chromatography under different temperature conditions. At the same time, by comparing the particle sizes of the whitening nano-compositions in each example before and after testing, it can be found that the average particle sizes of the whitening nano-compositions in each example after being placed at different temperature conditions for 3 months did not change obviously compared with before testing, which fully proved that the whitening nano-composition of the present application had good storage stability.

[0124] 2. Chicken embryo chorioallantoic membrane irritation test

[0125] After the whitening nano-compositions in Examples 1-5 were diluted 10 times with water, 200 μL was added to the surface of the chicken embryo chorioallantoic membrane, and after waiting for 300 s, the blood vessel changes were observed and the data were recorded, and the irritation score IS was calculated. The test results are shown in Figure 1. After the whitening nano-compositions diluted 10 times with water were contacted with the chicken embryo chorioallantoic membrane for 300 s, there was no bleeding, no blood vessel fusion, and no blood clotting phenomenon, and the irritation scores of Examples 1-5 were 0.07, which showed that the whitening nano-composition of the present application had good safety and no irritation.

[0126] 3. Patch test

[0127] 30 subjects were selected and randomly divided into 6 groups (5 people in each group), with a blank essence (2.0wt% glycerol, 0.3wt% carbomer, 0.1wt% xanthan gum, 0.5wt% phenoxyethanol 0.5% and the balance of purified water) as a blank control group, and the mixtures of the whitening nano-compositions in Examples 1-5 and the blank essence as experimental groups (the mass fraction of the whitening nano-composition relative to the blank essence was 10%); then the experimental groups and the blank control group were attached to the flexor of the forearm of the subjects (for 24 h), and after the indentation disappeared, the skin reaction was observed. The test results showed that none of the 30 subjects had pale erythema, erythema, edematous erythema, significant redness, infiltration or papules, and papules or blisters, etc., which showed that the whitening nano-composition of the present application had no irritation to human skin.

[0128] 4. Cumulative permeation and skin storage test

[0129] A blank serum was prepared with 2.0 wt% of glycerin, 0.3 wt% of carbomer, 0.1 wt% of xanthan gum, 0.5 wt% of phenoxyethanol and the rest of purified water. A whitening nano-composition serum (blank serum + 5% of the whitening nano-composition of Example 1 relative to the mass of the blank serum) and a whitening free-composition serum (blank serum + 5% of the whitening free-composition of Comparative Example 8 relative to the mass of the blank serum) were prepared. The Franz diffusion cell method was used to perform a transdermal experiment on isolated pig skin, with phosphate buffer PBS (pH = 7.4) as the receiving liquid, and stirring diffusion at 32°C. At 4, 8, 12 and 24 h, 0.5 mL of the receiving liquid was taken and an equal amount of fresh receiving liquid at constant temperature was immediately supplemented. High performance liquid chromatography (HPLC) was used for analysis, and the cumulative permeation amount per unit area of a- arbutin in the above-mentioned whitening nano-composition serum and whitening free-composition serum in the receiving liquid at different times was calculated. After 24 h, the skin was removed, washed, cut into pieces, ground into a homogenate, centrifuged with an appropriate amount of solvent, and the supernatant was analyzed by HPLC to calculate the skin storage amount per unit area of a- arbutin.

[0130] (1) Skin cumulative permeation amount

[0131] The skin cumulative permeation amount Q of a- arbutin at different sampling times was calculated according to the following formula.

[0132] Q = Q n / S, where S is the area of the diffusion cell 2.27 cm 2 , Q n is calculated according to the following formula:

[0133] In the above formula, Q n is the cumulative permeation amount of a- arbutin, C n is the drug concentration measured at the nth time, C i is the drug concentration measured at the ith point, V0 is the volume of the diffusion cell, i.e. the amount of receiving liquid added, and V i is the sampling amount each time.

[0134] (2) Skin storage amount

[0135] The skin storage amount Q m of a- arbutin at different sampling times was calculated according to the following formula.

[0136] Q m = C m × V m / S, where S is the area of the diffusion cell 2.27 cm 2 , C m is the drug concentration measured in the skin homogenate, and V m is the volume of the skin homogenate.

[0137] The test results of the skin cumulative permeation amount are shown in Figure 2. As shown in Figure 2, the 12h unit area skin cumulative permeation amount of a-arbutin in the whitening free composition serum and the whitening nano composition serum is 8.16 μg / cm 2 , 29.47 μg / cm 2 , respectively, and the 24h unit area skin cumulative permeation amount is 24.68 μg / cm 2 , 98.42 μg / cm 2 , respectively; that is, compared with the whitening free composition serum, the 12h unit area skin cumulative permeation amount of a-arbutin in the whitening nano composition serum is increased by 261.2%, and the 24h unit area skin cumulative permeation amount is increased by 298.8%. The test results of the skin storage amount are shown in Figure 3. As shown in Figure 3, the skin storage amount of a-arbutin in the whitening free composition serum and the whitening nano composition serum is 43.57 μg / cm 2 and 287.35 μg / cm 2 , respectively; that is, compared with the whitening free composition serum, the skin storage amount of a-arbutin in the whitening nano composition serum is increased by 261.2%. It can be seen that the whitening nano composition of the present application can effectively promote the skin penetration and storage of a-arbutin in the skin, thereby effectively improving the skin bioavailability thereof.

[0138] 5. Laser confocal microscope observation of skin penetration behavior

[0139] The sample after the test of the above-mentioned "4. Cumulative permeation amount and skin storage amount test" is labeled with RhoB and frozen section, and the skin tissue penetration behavior of free RhoB (the sample corresponding to the whitening free composition serum) and RhoB nano carrier (the sample corresponding to the whitening nano composition serum) at different time points is observed by laser confocal microscope, and the test results are shown in Figures 4 and 5.

[0140] As shown in Figures 4 and 5, the fluorescence penetration depth of the skin increases with the extension of time, the depth of free RhoB into the skin is 63.9 μm at 4h, and the depth of RhoB nano carrier into the skin is 305.6 μm, indicating that the whitening nano composition serum can penetrate into the deep tissue of the skin. In the same time, the relative fluorescence intensity of free RhoB is 3.8 at 4h, and the relative fluorescence intensity of RhoB nano carrier reaches 47.6, which is 11.53 times higher than that of free RhoB. That is, the fluorescence intensity of RhoB nano carrier in the skin is significantly stronger than that of free RhoB, further proving that the active nano carrier can effectively promote the transdermal absorption of the active ingredient to the target site of the skin, thereby improving the skin bioavailability of the whitening nano composition.

[0141] 6. Intracellular tyrosinase activity

[0142] The intracellular tyrosinase activity was determined by L-Dopa oxidation method, and the specific steps were as follows:

[0143] The B16F10 cells in logarithmic growth phase were inoculated in 24-well plates at a density of 5.0 x 10 4 After 24 h of culture, the cells were divided into a blank control group (only DMEM complete medium was added), a model group (only DMEM complete medium containing 100 nmol / L α-melanocyte stimulating hormone (α-MSH) was added), and an experimental group (DMEM complete medium containing 100 nmol / L α-MSH and 2000-fold diluted whitening nano-compositions in Comparative Examples 1 to 7, whitening free compositions in Comparative Example 8, and whitening nano-compositions in Example 1 was added), and three replicate wells were set in each group.

[0144] After 48 h of continuous culture, 200 μL of cell lysis buffer containing 1% (v / v) polyethylene glycol octylphenyl ether (Triton X-100) was added to each well, and the cells were frozen and lysed at -80°C for 30 min. The cell lysis mixture was collected by centrifugation, 100 μL of supernatant was taken to a 96-well plate, 100 μL of 0.1% (w / v) L-dopa solution was added, and the mixture was incubated at 37°C for 2 h. The absorbance (A) of each sample was measured at a wavelength of 495 nm using an enzyme marker, and the test results are shown in FIG. 6.

[0145] As can be seen from FIG. 6, compared with the model group, the whitening nano-compositions in Comparative Examples 1, 2, 3, 4, and 6, the whitening free compositions in Comparative Example 8, and the whitening nano-compositions in Example 1 can significantly reduce the tyrosinase activity (P < 0.05 or P < 0.01), and the whitening nano-compositions in Comparative Examples 5 and 7 have no significant effect on reducing the tyrosinase activity (P > 0.05), indicating that retinol palmitate is difficult to effectively inhibit the tyrosinase activity.

[0146] Compared with Comparative Example 6, the whitening nano-composition in Example 1 has a more significant effect on reducing the tyrosinase activity of B16F10 cells (P<0.01). Specifically, the whitening nano-composition consisting of a- arbutin, nicotinamide, 3-o-ethyl ascorbic acid and asiaticoside extract (Comparative Example 6) has an inhibition rate of 32.3% on the tyrosinase activity of B16F10 cells; the whitening nano-composition consisting of a- arbutin, nicotinamide, 3-o-ethyl ascorbic acid, asiaticoside extract and retinol palmitate (Example 1) has an inhibition rate of 64.7% on the tyrosinase activity of B16F10 cells, i.e. the inhibition rate of tyrosinase activity is increased by 100.3%. It can be seen that retinol palmitate has a synergistic effect with a- arbutin, nicotinamide, 3-o-ethyl ascorbic acid and asiaticoside extract, and can significantly inhibit the tyrosinase activity in cells.

[0147] Compared with Comparative Example 8 (inhibition rate of tyrosinase activity is 34.4%), the whitening nano-composition in Example 1 has a more significant effect on reducing the tyrosinase activity of B16F10 cells (P<0.01), and the inhibition rate of tyrosinase activity is increased by 88.1%, which fully demonstrates that the nano-carrier can enhance the inhibition rate of tyrosinase activity of active ingredients, and thus significantly improve the whitening effect of the whitening nano-composition.

[0148] 7. Determination of cell melanin content and observation of generation

[0149] The content of melanin in cells was determined by NaOH lysis method, and the specific steps are as follows:

[0150] B16F10 cells in the logarithmic growth phase were inoculated in a 24-well plate at a density of 5.0x10 4 The cells were divided into a control group (only DMEM complete medium was added), a model group (only DMEM complete medium containing 100 nmol / L a-MSH was added) and an experimental group (DMEM complete medium containing 100 nmol / L a-MSH and 2000-fold diluted whitening nano-compositions in Comparative Examples 1-7, whitening free composition in Comparative Example 8 and whitening nano-composition in Example 1 was added), and three replicate wells were set in each group.

[0151] After 48h of continuous culture, the cells in each well were collected in a centrifuge tube, 200μL of 1 mol / L NaOH solution containing 10% DMSO was added to each centrifuge tube, and then the EP (Eppendorf) tube was placed in a 80℃ constant temperature water bath for 1h of heating, to lyse the cells and dissolve the melanin, and then centrifuged, and the supernatant was taken to a 96-well plate, and the absorbance (A) of each well sample was measured at a wavelength of 405nm by an enzyme-labeled instrument, and the test results are shown in FIG. 7.

[0152] It can be found from Figure 7 that the whitening nano-composition in Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4 and Comparative Example 6, the whitening free composition in Comparative Example 8 and the whitening nano-composition in Example 1 can all significantly reduce the intracellular melanin content (P < 0.05 or P < 0.01) compared with the model group, and the whitening nano-compositions in Comparative Example 5 and Comparative Example 7 have no significant effect on reducing the intracellular melanin content (P > 0.05), indicating that retinol palmitate is difficult to effectively inhibit the intracellular melanin production.

[0153] Compared with Comparative Example 6, the whitening nano-composition in Example 1 has a more significant effect on reducing the intracellular melanin content of B16F10 cells (P < 0.01). Specifically, the whitening nano-composition consisting of α-arbutin, nicotinamide, 3-o-ethyl ascorbic acid and asiaticoside extract (Comparative Example 6) has an intracellular melanin content inhibition rate of 24.6% for B16F10 cells; the whitening nano-composition consisting of α-arbutin, nicotinamide, 3-o-ethyl ascorbic acid, asiaticoside extract and retinol palmitate (Example 1) has an intracellular melanin content inhibition rate of 56.8% for B16F10 cells, i.e. the intracellular melanin content inhibition rate is increased by 130.9%. It can be seen that retinol palmitate has a synergistic effect with α-arbutin, nicotinamide, 3-o-ethyl ascorbic acid and asiaticoside extract, and can significantly reduce the intracellular melanin content. At the same time, compared with Comparative Example 8, the intracellular melanin content inhibition rate of Example 1 is increased by 111.2%, i.e. it is fully demonstrated that retinol palmitate has a synergistic effect with α-arbutin, nicotinamide, 3-o-ethyl ascorbic acid and asiaticoside extract, can significantly promote the effect of the whitening active ingredients, and the nano-carrier can further enhance the effect of the whitening active ingredients.

[0154] 8. Human efficacy evaluation test

[0155] The blank essence is composed of 2.0 wt% of glycerol, 0.3 wt% of carbomer, 0.1 wt% of xanthan gum, 0.5 wt% of phenoxyethanol 0.5% and the balance of purified water.

[0156] Control group 1: blank essence;

[0157] Control group 2: blank essence + 5% of the whitening nano-composition in Comparative Example 1 relative to the mass fraction of the blank essence;

[0158] Control group 3: blank essence + 5% of the whitening nano-composition in Comparative Example 2 relative to the mass fraction of the blank essence;

[0159] Control group 4: blank essence + 5% of the whitening nano-composition in Comparative Example 3 relative to the mass fraction of the blank essence;

[0160] Control group 5: blank serum + 5% of the whitening nano-composition in Comparative Example 4 relative to the mass fraction of the blank serum;

[0161] Control group 6: blank serum + 5% of the whitening nano-composition in Comparative Example 5 relative to the mass fraction of the blank serum;

[0162] Control group 7: blank serum + 5% of the whitening nano-composition in Comparative Example 6 relative to the mass fraction of the blank serum;

[0163] Control group 8: blank serum + 5% of the whitening nano-composition in Comparative Example 7 relative to the mass fraction of the blank serum;

[0164] Control group 9: blank serum + 5% of the whitening free composition in Comparative Example 8 relative to the mass fraction of the blank serum;

[0165] Experimental group: blank serum + 5% of the whitening nano-composition in Example 1 relative to the mass fraction of the blank serum.

[0166] Thirty experimental volunteers were recruited, aged between 20-60 years old, healthy, without skin diseases, non-pregnant or lactating, and no allergy to product ingredients. After being informed of the evaluation process, possible effects, risks, precautions, and signing the efficacy evaluation informed consent form, they were randomly divided into 10 groups (3 people in each group) for efficacy evaluation test of the above control groups and experimental group. Specifically, the samples were used on the face twice a day at regular intervals, and no other products or drugs that could affect the skin condition were used during the test. The skin condition and feelings after use were recorded and fed back accurately. After 4 weeks (28 days) of sample application, the volunteers' efficacy indicators were observed and compared. The skin color and uniformity (ITA° value), skin gloss (L* value of CIELab* color space (brightness)), total area of melanin and pigmented spots, melanin index (MI value) of the face were determined by Shanghai Fuhuan Intelligent Skin Analysis System and German CK Skin Melanin Tester MX18. The changes in each index parameter after using the sample were calculated and the mean value was taken.

[0167] Table 2 Change rate of each skin index of volunteers before and after using the sample

[0168] According to the data in Table 2, after the volunteers used the sample of control group 1 (blank serum) for 4 weeks, the ITA° value increased by 0.31%, the skin L* value increased by 0.22%, the MI value decreased by 0.54%, and the melanin pigmented spot area decreased by 0.15%, i.e. there was no significant change in the above indicators, indicating that the blank serum had no improvement effect on skin color and uniformity, melanin and pigmented spots.​

[0169] Compared with the control group 1, the volunteers corresponding to the control groups 2-7, the control group 9 and the experimental group had obvious improvements in the ITA° value and the L* value of the skin, and significant reductions in the MI value and the total area of the melanin spots after using the samples for 4 weeks; the control group 8 had a limited improvement in the whitening effect due to the low content of retinol palmitate. Among them, the sample of the control group 7 (the whitening nano composition composed of α-arbutin, niacinamide, 3-o-ethyl ascorbic acid and the extract of G. japonica) had a significantly improved effect, the ITA° value of the skin of the corresponding volunteer increased by 6.58%, the L* value increased by 5.16%, the MI value decreased by 8.47%, and the total area of the melanin spots decreased by 10.28%; and the sample of the experimental group (the whitening nano composition composed of α-arbutin, niacinamide, 3-o-ethyl ascorbic acid and the extract of G. japonica and retinol palmitate + the blank serum) had a better effect, after the corresponding volunteer used the sample, the ITA° value of the skin increased by 11.08%, the L* value increased by 11.43%, the MI value decreased by 18.67%, and the total area of the melanin spots decreased by 21.42%, which fully illustrated that α-arbutin, niacinamide, 3-o-ethyl ascorbic acid, the extract of G. japonica and retinol palmitate had a synergistic whitening effect.

[0170] In addition, after using the sample of the control group 9 (the whitening free composition in Comparative Example 8 + the blank serum), the ITA° value of the skin of the corresponding volunteer increased by 6.86%, the L* value increased by 5.96%, the MI value decreased by 9.15%, and the total area of the melanin spots decreased by 10.76%, i.e. the whitening effect was weaker than that of the experimental group, and it could be seen that the combination of the whitening active ingredients and the nano carrier could effectively improve the skin color and uniformity, the melanin and the spots.

[0171] FIGS. 8 and 9 are the face color contrast diagrams (FIG. 8A, FIG. 9A) and the face pigment and spot contrast diagrams (FIG. 8B, FIG. 9B) of the different volunteers (volunteers 1 and 2) using the sample of the experimental group on the 0th day, the 14th day and the 28th day, which were collected by the Vplus device. As can be seen from FIGS. 8 and 9, after using the serum containing 5% of the whitening nano composition in Example 1 for 28 days, the number and the average area of the melanin and the spots on the face of the volunteer were reduced, the uniformity and the gloss of the skin color were obviously improved, and the sample had an excellent whitening effect.

[0172] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present application, but not to limit the protection scope of the present application. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can be modified or replaced equivalently without departing from the essence and scope of the technical solutions of the present application.

Claims

1. A whitening nano-composition, characterized in that, The active ingredient is coated and / or adsorbed by the nanocarrier; The active ingredient comprises, by weight fraction, the following components: 1-15 parts of alpha-arbutin, 1-15 parts of nicotinamide, 1-10 parts of 3-o-ethyl ascorbic acid, 0.1-1 part of centella asiatica extract, and 1-5 parts of retinol palmitate; The nanocarrier comprises, by weight fraction, the following components: 5-10 parts of emulsifier, 5-20 parts of polyol, 0.1-5 parts of phospholipid, and 40-60 parts of water.

2. The whitening nano-composition of claim 1, wherein, The mass ratio of the active ingredient to the nanocarrier is (15-30):(65-80).

3. The whitening nano-composition of claim 1 or 2, wherein, The emulsifier comprises at least one of polyoxyethylene castor oil emulsifier, polyoxyethylene hydrogenated castor oil emulsifier, polyglycerol emulsifier, poloxamer, and cocoglycoside.

4. The whitening nano-composition of claim 1 or 2, wherein, The polyol comprises at least one of glycerol, propylene glycol, butylene glycol, 1,3-propanediol, 1,2-pentanediol, 1,2-hexanediol, dipropylene glycol, isopropyl alcohol, polyethylene glycol-200, PPG-10 sorbitol, and octyldodecanol.

5. The whitening nanocomposition as described in claim 1 or 2, characterized in that, The phospholipid comprises at least one of hydrogenated lecithin, lecithin, and soybean lecithin.

6. A process for the preparation of the whitening nano-composition according to any one of claims 1 to 5, characterized in that, The method comprises the following steps: S1, uniformly mixing alpha-arbutin, nicotinamide, 3-o-ethyl ascorbic acid, and centella asiatica extract with phospholipid, water, and a part of polyol to obtain a mixed solution I; S2, uniformly mixing retinol palmitate with emulsifier and the remaining polyol to obtain a mixed solution II; S3, uniformly mixing the mixed solution I in S1 with the mixed solution II in S2, homogenizing, and obtaining the whitening nanocomposition.

7. The method for preparing the whitening nanocomposition as described in claim 6, characterized in that, At least one of the following (I)-(III): (I) The step S1 specifically comprises stirring alpha-arbutin, nicotinamide, 3-o-ethyl ascorbic acid, and centella asiatica extract with phospholipid, water, and a part of polyol at 40-65°C and 150-200 rpm for 10-30 min to obtain the mixed solution I; (II) The step S2 specifically comprises stirring retinol palmitate with emulsifier and the remaining polyol at 40-65°C and 150-200 rpm for 10-30 min to obtain the mixed solution II; (III) The step S3 specifically comprises stirring the mixed solution I in S1 with the mixed solution II in S2 at 40-65°C and 200-270 rpm for 10-30 min, homogenizing, and obtaining the whitening nanocomposition.

8. Use of the whitening nanocomposition of any one of claims 1-5 in the preparation of skin care or cosmetic products.

9. A whitening skin care or whitening cosmetic product comprising the whitening nanocomposition of any one of claims 1-5.

10. The whitening skin care or whitening cosmetic product according to claim 9, wherein The mass fraction of the whitening nanocomposition in the whitening skin care or whitening cosmetic product is 0.1%-30%.

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