Mild and stable composition with whitening and anti-aging effects as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biology. More specifically, the invention relates to a mild and stable composition with whitening and anti-aging effects, an emulsion and a preparation method and application thereof. The emulsion is prepared from the following components in percentage by mass: 0.1 to 1.5 percent of hydroxydecyl ubiquinone, 0.1 to 1 percent of leuconostoc / radish root fermentation product filtrate, 0.01 to 1 percent of 2 '-fucosyllactose, 1 to 5 percent of an emulsifier, 0.1 to 0.3 percent of a chelating agent, 0.1 to 0.5 percent of a thickening agent, 5 to 20 percent of an alcohol agent and the balance of ultrapure water. According to the emulsion disclosed by the invention, the leuconostoc / radish root fermentation product filtrate is matched with the 2 '-fucosyllactose, so that the irritation of the hydroxydecyl ubiquinone is greatly reduced, the stability of the hydroxydecyl ubiquinone is improved, and the prepared emulsion contains the hydroxydecyl ubiquinone with relatively high concentration and is stable in performance and low in irritation.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and more specifically, relates to a mild and stable composition with whitening and anti-aging effects, and a preparation method and application thereof. Background Art

[0002] Hydroxydecylubiquinone (HYDROXYDECYL UBIQUINONE), also known as idebenone (structural formula shown in Formula 1 below), is a highly effective antioxidant that has the effects of scavenging free radicals, inhibiting lipid peroxidation, inhibiting inflammation, inhibiting DNA damage, photoprotection, and reducing pigmentation. Hydroxydecylubiquinone is an antioxidant developed based on the structure of coenzyme Q10. It is a coenzyme Q10 analog with a low relative molecular mass. Its side chain is shorter, there is no isoprene unit, and an alcohol hydroxyl group is added to the end of the side chain, which makes it easier to pass through biological membranes and blood-brain barriers, and has stronger antioxidant activity than coenzyme Q10.

[0003]

[0004] Idebenone was first developed by Japan's Takeda Pharmaceutical Company and was launched on the market in 1986. It can improve brain glucose utilization and improve brain dysfunction by activating mitochondrial function. It also plays a strong antioxidant and free radical scavenging role. In 2005, the pharmaceutical company Allergan discovered that idebenone has antioxidant effects when used topically on the skin, so it reached a cooperation with the high-end American skincare brand Elizabeth Arden to jointly develop products containing 0.5% and 1.0% idebenone. Although idebenone has excellent anti-aging effects, many contact dermatitis patients use skincare products containing idebenone. [1] ; In addition, Amorim et al. [2] The mucosal irritation experiment using L929 cells found that 1% idebenone showed a similar degree of irritation as the positive control (1% SDS). The concentration of idebenone in Elizabeth Arden Prevage is also only 0.5%, and it has added soothing ingredients such as green tea and ceramide to prevent allergies. Some scholars have compared the local safety and effectiveness of 0.5% and 1.0% idebenone on photodamaged skin. The results showed that after using the 1.0% idebenone formula for six weeks, the degree of skin roughness / dryness, the reduction of fine lines / wrinkles, and the overall improvement of photodamaged skin were all higher than those of the 0.5% concentration. [3] Secondly, Amorim et al. [2] After being stored at 40°C and 75% humidity for 45 days, idebenone degraded by 60%, its antioxidant activity decreased by 30%, and its chemical stability was poor. Therefore, in the prior art, idebenone is usually prepared into liposomes to improve its stability, but its irritation is not greatly improved.

[0005] [1]IBOGDAN ALLEMANNMD,L.BAUMANNMD.Antioxidants Usedin Skin CareFormulations[J].Skin TherapyLett,2008,13(7):5-9.

[0006] [2]AMORIMCM, COUTOAG, NETzDJA, eta1. Antioxidant Idebenone-load edNanoparticles Based on Chitosan and N-carboxymethylchitosan[J]. Nano-medicine, 2010, (6): 745-752.

[0007] [3] Yu Xiao, https: / / www.zhihu.com / question / 38982132 / answer / 2030368181. Summary of the invention

[0008] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of existing idebenone products with a concentration greater than 0.5%, such as strong irritation to the skin and stability, and to provide a high-concentration and mild idebenone-containing product.

[0009] The purpose of the present invention is to provide a mild and stable emulsion with whitening and anti-aging effects.

[0010] Another object of the present invention is to provide a method for preparing the emulsion.

[0011] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0012] In one embodiment, a mild and stable emulsion with whitening and anti-aging effects is provided, wherein the emulsion is prepared from the following components in mass fractions: 0.1-1.5% hydroxydecylubiquinone, 0.1-1% filtrate of leuconostoc / radish root fermentation product, 0.01-1% 2'-fucosyllactose, 1-5% emulsifier, 0.1-0.3% chelating agent, 0.1-0.5% thickener, 5-20% alcohol agent and the remainder ultrapure water.

[0013] In one embodiment, the weight ratio of the Leuconostoc / radish root fermentation product filtrate to 2'-fucosyllactose is 8-15:1.

[0014] In one embodiment, the weight ratio of the Leuconostoc / radish root fermentation product filtrate to 2'-fucosyllactose is 10:1.

[0015] In one embodiment, the emulsifier is one or more of caprylic triglyceride, capric triglyceride, PPG-26 buteth-26, polyglyceryl 10 stearate and PEG-40 hydrogenated castor oil.

[0016] In one embodiment, the thickener is one or more of carbomer, hydroxyethyl cellulose, chitosan, hydroxypropyl methyl cellulose, sclerotium gum and yeast β-glucan.

[0017] In one embodiment, the alcohol agent is one or more of glycerol, 1,2-propylene glycol, 1,3-propylene glycol, 1,2-pentanediol and 1,2-hexanediol.

[0018] In one embodiment, the chelating agent is disodium ethylenediaminetetraacetate.

[0019] In one embodiment, the present invention provides a mild and stable emulsion with whitening and anti-aging effects, which is prepared from the following components in mass fractions: 0.1-1.5% hydroxydecylubiquinone, 0.1-1% filtrate of leuconostoc / radish root fermentation product, 0.01-1% 2'-fucosyllactose, 1-3% PPG-26 butanol polyether-26, 1-2% PEG-40 hydrogenated castor oil, 0.1-0.3% disodium ethylenediaminetetraacetic acid, 0.1-0.5% carbomer, 5-20% 1,3-propylene glycol and the remainder of ultrapure water.

[0020] In one embodiment, the present invention provides a method for preparing the emulsion, comprising the following steps:

[0021] S1. Add hydroxydecylubiquinone to 1,3-propylene glycol and perform ultrasonication at room temperature for 10 to 20 minutes to obtain phase A.

[0022] S2, mixing carbomer, disodium ethylenediaminetetraacetic acid, filtrate of Leuconostoc / radish root fermentation product, 2'-fucosyllactose and ultrapure water, stirring at 100-300 rpm for 10-30 min, and heating to 70-85° C. to obtain phase B;

[0023] S3, mix PPG-26 buteth-26 and PEG-40 hydrogenated castor oil, heat to 70-85° C., stir and dissolve, and obtain phase C;

[0024] S4: Slowly add phase A into phase B, stir at 40-55°C for 20-60 min, then gradually drop it into phase C under shearing conditions, homogenize and emulsify for 3-8 min, cool to room temperature, and obtain an emulsion.

[0025] In one embodiment, the shear speed is 6500-7500 r / min, the shear temperature is 45-55°C, and the shear time is 3-8 min; the homogenization conditions are: pressure 200-400 bar, temperature 55-80°C, and cycle 2-4 times.

[0026] Leuconostoc / Radish Root Ferment Filtrate, INCI name: Leuc onostoc / Radish Root Ferment Filtrate, is a natural preservative derived from Leuconostoc used for fermenting kimchi. The bacterium forms peptides (Bac teriocins / Antimicrobial Peptides) with antimicrobial activity during the fermentation process, which have strong killing and inhibitory effects on Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Aspergillus niger, Klebsiella pneumoniae, and Burkholderia cepacia. It is suitable for a wide range of pH values ​​and is generally used as a substitute for chemical preservatives in the art. Before the application date, it has not been found that it has the effect of reducing the irritation of idebenone.

[0027] 2'-fucosyllactose (2'-FL) is the most abundant functional oligosaccharide in breast milk. It has physiological functions such as regulating the intestinal environment of infants, enhancing intestinal immunity, and promoting the development of infant brain nerves. It has been widely used in infant formula and dietary supplements. Before the application date, it had not been found to reduce the irritation of idebenone.

[0028] However, the present invention found that a specific concentration of Leuconostoc / radish root fermentation product filtrate combined with 2'-fucosyllactose can greatly reduce the irritation of idebenone, and the stability and antioxidant properties of the prepared emulsion are also improved. A single test result shows that this effect is produced after the combination. DETAILED DESCRIPTION

[0029] The present invention can be further described by the following examples, however, the scope of the present invention is not limited to the following examples. It will be appreciated by those skilled in the art that various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention.

[0030] Example 1

[0031] A mild and stable emulsion with whitening and anti-aging effects is prepared from the following components in mass fractions: 1% hydroxydecylubiquinone, 1% leuconostoc / radish root fermentation product filtrate, 0.1% 2'-fucosyllactose, 2% PPG-26 buteth-26, 1% PEG-40 hydrogenated castor oil, 0.1% disodium edetate, 0.5% carbomer, 15% 1,3-propylene glycol and the balance ultrapure water.

[0032] Preparation method:

[0033] S1, adding hydroxydecylubiquinone to 1,3-propylene glycol, and ultrasonicating at room temperature for 15 min to obtain phase A;

[0034] S2, mixing carbomer, disodium ethylenediaminetetraacetic acid, filtrate of Leuconostoc / radish root fermentation product, 2'-fucosyllactose and ultrapure water, stirring at 200 rpm for 25 min, and heating to 80° C. to obtain phase B;

[0035] S3, mixing PPG-26 buteth-26 and PEG-40 hydrogenated castor oil, heating to 80° C. and stirring to dissolve, to obtain phase C;

[0036] S4. Slowly add phase A into phase B, stir at 50°C for 30 minutes, then gradually drop it into phase C under shearing conditions, homogenize and emulsify for 5 minutes, cool to room temperature, and obtain an emulsion; wherein the shear speed is 7000r / min, the shear temperature is 50°C, and the shear time is 6 minutes; the homogenization conditions are: pressure 300bar, temperature 65°C, and cycle 3 times.

[0037] Example 2

[0038] A mild and stable emulsion with whitening and anti-aging effects is prepared from the following components in mass fractions: 1% hydroxydecylubiquinone, 0.8% leuconostoc / radish root fermentation product filtrate, 0.1% 2'-fucosyllactose, 2% PPG-26 buteth-26, 1% PEG-40 hydrogenated castor oil, 0.1% disodium edetate, 0.5% carbomer, 15% 1,3-propylene glycol and the balance ultrapure water.

[0039] The preparation method is similar to that of Example 1.

[0040] Example 3

[0041] A mild and stable emulsion with whitening and anti-aging effects is prepared from the following components in mass fractions: 1% hydroxydecylubiquinone, 0.6% leuconostoc / radish root fermentation product filtrate, 0.04% 2'-fucosyllactose, 2% PPG-26 buteth-26, 1% PEG-40 hydrogenated castor oil, 0.1% disodium edetate, 0.5% carbomer, 15% 1,3-propylene glycol and the balance ultrapure water.

[0042] The preparation method is similar to that of Example 1.

[0043] Example 4

[0044] A mild and stable emulsion with whitening and anti-aging effects is prepared from the following components in mass fractions: 1% hydroxydecylubiquinone, 1% leuconostoc / radish root fermentation product filtrate, 0.1% 2'-fucosyllactose, 3% polyglycerol 10 stearate, 2% PEG-40 hydrogenated castor oil, 0.1% disodium edetate, 0.3% chitosan, 15% 1,2-hexanediol and the balance ultrapure water.

[0045] The preparation method is similar to that of Example 1.

[0046] Example 5

[0047] A mild and stable emulsion with whitening and anti-aging effects is prepared from the following components in mass fractions: 1% hydroxydecylubiquinone, 1% leuconostoc / radish root fermentation product filtrate, 0.1% 2'-fucosyllactose, 3% caprylic triglyceride, 2% PEG-40 hydrogenated castor oil, 0.1% disodium ethylenediaminetetraacetic acid, 0.3% chitosan, 15% 1,2-hexanediol and the balance ultrapure water.

[0048] The preparation method is similar to that of Example 1.

[0049] Effects of different compositions on transepidermal water loss (TEWL) of damaged skin

[0050] Test method: 10 people who are sensitive to hydroxydecylubiquinone were screened, aged between 20 and 35 years old, with an average age of 26.43±1.27 years old. Emulsion samples A1-A8 (Table 1 below) containing different proportions of leuconostoc / radish root fermentation product filtrate and 2'-fucosyllactose were prepared according to the formula of Example 1. Before the test, the inner side of the forearm of the test subject was rinsed with clean water, wiped clean with a paper towel, and waited for 30 minutes in a constant temperature and humidity environment (temperature 20-22°C, humidity 40%-60%). The inner side of the left forearm of each test subject was divided into 4 square areas, and the inner side of the right forearm was divided into 4 square areas, each area was 2×2cm in size, and the TEWL value before applying the sample in each area was tested using a skin epidermal water loss tester (Tewameter TM210, C+K Company, Germany), which was recorded as the skin TEWL value of the test subject before using the sample. The A1-A8 emulsion samples shown in Table 1 were respectively applied to 8 square areas on the subjects' arms (1 g of the emulsion sample was taken and smeared in circles until it was completely absorbed, once in the morning and evening, for 2 consecutive days). The changes in TEWL values ​​of each smeared area were tested on the 3rd day, and the TEWL difference (TEWL difference after use - TEWL difference before use) was calculated. The data were statistically analyzed to compare the differences among the groups. The results are shown in Table 2.

[0051] Table 1A1-A8 emulsion mass ratio

[0052]

[0053] Note: RS is the abbreviation for Leuconostoc / radish root fermentation filtrate.

[0054] Table 2 Comparison of TEWL differences before and after use of different compositions

[0055]

[0056]

[0057] As can be seen from Table 2, after using the lotion sample A1 containing only hydroxydecylubiquinone, the TEWL value of the subjects increased significantly, indicating that hydroxydecylubiquinone can irritate the skin and significantly damage the skin barrier; after using the lotion sample containing the filtrate of the fermentation product of Leuconostoc / radish root A2 or the hydroxydecylubiquinone containing 2'-fucosyllactose A3, the TEWL value of the skin of the subjects also increased, but the increase was less than that of sample A1; but after using the hydroxydecylubiquinone samples A4 to A8 containing the filtrate of the fermentation product of Leuconostoc / radish root and 2'-fucosyllactose, the TEWL value of the skin of the subjects became significantly smaller than before use, especially when the ratio of the two was in the range of 8 to 15:1. This shows that the combination of the two can reduce the irritation of hydroxydecylubiquinone to the skin, repair the damaged skin barrier caused by it, and the combination has a synergistic effect within a specific ratio range.

[0058] Stability test

[0059] Samples A1 to A8 were placed in a constant temperature environment of 4°C and 45°C for 90 days respectively, and then taken out to recover to room temperature for 30 minutes and then observed for the properties of the samples to determine whether oil-water stratification, agglomeration and precipitation occurred. The results are shown in Table 3 below.

[0060] Table 3 Stability test results

[0061]

[0062]

[0063] It can be seen from Table 3 above that the samples containing the filtrate of the fermentation product of Leuconostoc / radish root and 2'-fucosyllactose have higher stability. After being placed in a constant temperature environment of 4°C and 45°C for 90 days, there is no oil-water stratification, agglomeration or precipitation. However, the samples without the above two components or containing only the filtrate of the fermentation product of Leuconostoc / radish root have poor stability, with a small amount of precipitation and slight stratification.

[0064] Human patch test

[0065] According to the "Human Patch Test" in the "Technical Specifications for Safety of Cosmetics" (2015 edition), 30 subjects were selected according to the subject selection criteria, and qualified patch materials were selected. The samples were placed in the patch device (the dosage was 0.02g), and the control well was blank. The inner side of the left forearm of each subject was divided into 5 square areas, and the inner side of the right forearm was divided into 4 square areas. The samples and blank controls were respectively pasted in 9 square areas, and the palms were gently pressed to make them evenly applied to the skin for 24 hours. The skin reaction was observed repeatedly at 30 minutes, 24 hours and 48 hours after the patch device was removed. The skin reaction was judged with reference to the internationally accepted four-level scoring standard (Table 4), and the observation results were recorded. The average irritation score (the sum of the score values ​​of 30 subjects / 30) was calculated. The results are shown in Table 5 below.

[0066] Table 4 Skin reaction scoring criteria

[0067]

[0068]

[0069] Table 5 Average stimulus integral value (VS)

[0070] Sample No. Patch 24h Patch 48h A1 0.167 0.133 A2 0.1 0.1 A3 0.067 0.083 A4 0 0 A5 0 0 A6 0 0 A7 0.017 0 A8 0.05 0.033

[0071] As can be seen from Table 5 above, the use of hydroxydecylubiquinone A4-A6 containing Leuconostoc / radish root fermentation product filtrate and 2'-fucosyllactose in a ratio of 8-15:1 has no adverse reaction to the skin. And there is a correlation between the VS value and TEWL value of each group of samples. The more severe the clinical erythema reaction, the higher the TEWL value.

[0072] Human epidermal cell antioxidant test

[0073] HaCaT cells were cultured in DMEM medium containing 10% FBS at 37°C, 5% CO2, and 95% relative humidity. The medium was changed every other day. When the cell density reached 80-90%, the cells were digested with 0.25% trypsin containing EDTA and passaged at a ratio of 1:3. Human epidermal cells HaCaT in the logarithmic growth phase were selected and the cell concentration was adjusted to 1×10 5 / mL, inoculated into a 96-well plate at 100μL / well, incubated for 24h, added 20μL of sample solution and incubated for 24h (A1~A4 were prepared with distilled water to prepare a sample solution with a concentration of 5μg / mL, the model group added an equal amount of culture medium without sample, and each sample was made into 6 duplicate wells), the culture solution was removed, 300μL of hydrogen peroxide was added after washing with PBS, and an equal amount of culture medium was added to the blank control group, the culture medium was removed after 4h, PBS was washed once, 20μL of 5mg / mL MTT solution was added to each well, incubated for 4h, the spacer solution was discarded, and 150μL of DMSO was added to each well to dissolve the crystals. The 96-well plate was placed on a flat shaker and mixed at low speed for 10min, and the OD value of each well was measured at a wavelength of 490nm using an enzyme marker. The relative cell viability was calculated, and the results are shown in Table 6 below.

[0074] Table 6 Effects of different sample treatments on relative viability of HaCaT cells (X±s, n=6)

[0075] Sample No. Relative cell viability / % Blank control group 100.00 Model Group <![CDATA[59.63±5.24 ** ]]> A1 <![CDATA[80.41±3.59 ## ]]> A2 <![CDATA[83.05±1.62 ## ]]> A3 <![CDATA[85.63±5.07 ## ]]> A4 <![CDATA[95.18±2.36 ###^^ ]]>

[0076] Note: Compared with the blank control group, ** P<0.01; compared with the model group, ## P < 0.01; ### P<0.001, compared with group A1, ^^ P<0.01.

[0077] It can be seen from Table 6 above that after HaCaT cells were damaged by hydrogen peroxide, the cell viability decreased to 59.63%, and after adding the sample solutions containing hydroxydecylubiquinone, the relative cell viability was significantly improved, especially the hydroxydecylubiquinone sample containing the filtrate of the fermentation product of Leuconostoc / radish root and 2'-fucosyllactose, the relative cell viability was the highest, which indicates that the addition of the filtrate of the fermentation product of Leuconostoc / radish root and 2'-fucosyllactose enhances the antioxidant effect of hydroxydecylubiquinone.

[0078] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A mild and stable emulsion with whitening and anti-aging effects, characterized in that: The emulsion is prepared from the following components in mass fractions: 0.1-1.5% hydroxydecylubiquinone, 0.1-1% filtrate of leuconostoc / radish root fermentation product, 0.01-1% 2'-fucosyllactose, 1-5% emulsifier, 0.1-0.3% chelating agent, 0.1-0.5% thickener, 5-20% alcohol agent and the balance of ultrapure water.

2. The emulsion according to claim 1, characterized in that The weight ratio of the filtrate of the Leuconostoc / radish root fermentation product to 2'-fucosyllactose is 8-15:

1.

3. The emulsion according to claim 2, characterized in that The weight ratio of the filtrate of the Leuconostoc / radish root fermentation product to 2'-fucosyllactose is 10:

1.

4. The emulsion according to any one of claims 1 to 3, characterized in that The emulsifier is one or more of caprylic triglyceride, capric triglyceride, PPG-26 buteth-26, polyglyceryl 10 stearate and PEG-40 hydrogenated castor oil.

5. The emulsion according to any one of claims 1 to 3, characterized in that The thickener is one or more of carbomer, hydroxyethyl cellulose, chitosan, hydroxypropyl methyl cellulose, sclerotium gum and yeast β-glucan.

6. The emulsion according to any one of claims 1 to 3, characterized in that The alcohol agent is one or more of glycerol, 1,2-propylene glycol, 1,3-propylene glycol, 1,2-pentanediol and 1,2-hexanediol.

7. The emulsion according to any one of claims 1 to 3, characterized in that The chelating agent is disodium ethylenediaminetetraacetate.

8. A mild and stable emulsion with whitening and anti-aging effects, characterized in that: The emulsion is prepared from the following components in mass fractions: 0.1-1.5% hydroxydecylubiquinone, 0.1-1% filtrate of leuconostoc / radish root fermentation product, 0.01-1% 2'-fucosyllactose, 1-3% PPG-26 butanol polyether-26, 1-2% PEG-40 hydrogenated castor oil, 0.1-0.3% disodium ethylenediaminetetraacetate, 0.1-0.5% carbomer, 5-20% 1,3-propylene glycol and the balance of ultrapure water.

9. A method for preparing the emulsion according to claim 8, characterized in that: The following steps are involved: S1. Add hydroxydecylubiquinone to 1,3-propylene glycol and perform ultrasonication at room temperature for 10 to 20 minutes to obtain phase A. S2, mixing carbomer, disodium ethylenediaminetetraacetic acid, filtrate of Leuconostoc / radish root fermentation product, 2'-fucosyllactose and ultrapure water, stirring at 100-300 rpm for 10-30 min, and heating to 70-85° C. to obtain phase B; S3, mix PPG-26 buteth-26 and PEG-40 hydrogenated castor oil, heat to 70-85° C., stir and dissolve, and obtain phase C; S4: Slowly add phase A into phase B, stir at 40-55°C for 20-60 min, then gradually drop it into phase C under shearing conditions, homogenize and emulsify for 3-8 min, cool to room temperature, and obtain an emulsion.

10. The preparation method according to claim 9, characterized in that: The shearing speed is 6500-7500 r / min, the shearing temperature is 45-55° C., and the shearing time is 3-8 min. The homogenizing conditions are: pressure 200-400 bar, temperature 55-80° C., and circulation 2-4 times.

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