Oral composition with whitening effect as well as product and application thereof

Abstract

The invention provides an oral composition with a whitening effect as well as a product and application thereof, and relates to the technical field of compositions. The composition is composed of glabridin and glutathione, and the mass ratio of glabridin to glutathione in the composition is 1: (1-200). The two components in the whitening composition have a synergistic effect, so that the generation of melanin can be reduced, the pigment content can be reduced, the melanin can be strongly inhibited on the premise of not influencing the normal cell activity of the skin, and a new effective scheme is provided for research and development of oral whitening products.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of compositions, and particularly relates to an oral whitening composition, a product thereof and application. BACKGROUND

[0002] The core cause of skin pigmentation (such as chloasma, sunburn, dullness) is abnormal activity of melanocytes. The synthesis of melanin starts from the conversion of tyrosine into dopa under the catalysis of tyrosinase (TYR), and then generates eumelanin and pheomelanin through the regulation of tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2). Oxidative stress and release of inflammatory factors (such as IL-6 and TNF-α) caused by ultraviolet irradiation can further activate the small eye malformation-related transcription factor signaling pathway of melanocytes, aggravate the synthesis and transport of melanin, and ultimately lead to skin pigmentation.

[0003] Current oral whitening ingredients have limitations in practical application. Vitamin C and its derivatives can play a whitening role by reducing dopaquinone and inhibiting tyrosinase activity, but they are easily degraded by gastrointestinal enzymes when taken orally, have low bioavailability, and need to be taken in large doses for a long time to show effects. Excessive intake may cause diarrhea, increased uric acid, and other problems. Whitening is achieved by blocking the activation of plasmin on melanocytes, but oral administration can cause gastrointestinal discomfort, and long-term use may affect blood clotting function, so the dose needs to be strictly controlled. Vitamin E focuses on indirectly reducing melanin production by antioxidant ROS scavenging, but the mechanism is single, the inhibition effect on activated melanocytes is weak, the whitening period is long, and the consumer compliance is low.

[0004] Chinese patent CN115337235A discloses a multiple whitening pathway composition containing safflower camellia extract. The composition includes the following components in mass fraction: dihydric alcohol 18-50 parts, glycerol 13-35 parts, polyoxyethylene sorbitan fatty acid ester 0.003-25 parts, hexylresorcinol 0.5-7 parts, glabriden 0.05-2 parts, paeonol 0.05-4 parts, xyloestryth extract 0.05-2 parts, VE acetate 0.05-2 parts, camellia flower and leaf extract 0.44-21 parts, water 9-32 parts, arbutin 0.5-7 parts, and nicotinamide 0.5-18 parts. The composition has good anti-glycation effect and achieves whitening effect through multiple pathways, but its whitening effect still needs to be improved.

[0005] In existing technologies, oral whitening compositions are mostly combinations of single active ingredients and absorption enhancers, aiming to improve the bioavailability of individual ingredients. Furthermore, the application of glycyrrhizin is limited to topical use, and no research has explored its potential oral combination with glutathione. Therefore, developing an oral whitening composition of glycyrrhizin-glutathione that can significantly improve whitening effects has become a key direction for overcoming the current bottlenecks in oral whitening technology. Summary of the Invention

[0006] To address the aforementioned problems, this invention provides a composition for skin whitening, its preparation method, and its application.

[0007] On one hand, the present invention provides an oral whitening composition, the composition being composed of glycyrrhizin and glutathione.

[0008] Specifically, the mass ratio of glycyrrhizin to glutathione in the composition is 1:1-200.

[0009] According to some embodiments of the present invention, the mass ratio of glutathione to glycyrrhizin in the composition is 1:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1, 85:1, 90:1, 95:1, or 100:1. The following ratios can be selected: 105:1, 110:1, 115:1, 120:1, 125:1, 130:1, 135:1, 140:1, 145:1, 150:1, 155:1, 160:1, 165:1, 170:1, 175:1, 180:1, 185:1, 190:1, 195:1, 200:1, and any value in between.

[0010] Furthermore, the mass ratio of glutathione to glycyrrhizin in the composition is 10-125:1.

[0011] According to some embodiments of the present invention, the mass ratio of glutathione to glycyrrhizin in the composition is 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1, 100:1, 110:1, 120:1, 125:1, and any intermediate value may be selected.

[0012] Furthermore, the mass ratio of glycyrrhizin to glutathione in the composition is 1:50-80.

[0013] Furthermore, the mass ratio of glycyrrhizin to glutathione in the composition is 1:63.

[0014] According to some embodiments of the present invention, the composition comprises 1-10 parts of glycyrrhizin and 1-200 parts of glutathione.

[0015] According to some embodiments of the present invention, the composition comprises, by weight, 1 part, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts, 5 parts, 5.5 parts, 6 parts, 6.5 parts, 7 parts, 7.5 parts, 8 parts, 8.5 parts, 9 parts, 9.5 parts, 10 parts, and any intermediate value may be selected.

[0016] According to some embodiments of the present invention, the composition comprises, by weight, 1 part, 10 parts, 20 parts, 30 parts, 40 parts, 50 parts, 60 parts, 70 parts, 80 parts, 90 parts, 100 parts, 110 parts, 120 parts, 130 parts, 140 parts, 150 parts, 160 parts, 170 parts, 180 parts, 190 parts, 200 parts of glutathione, and any intermediate value may be selected.

[0017] Secondly, the present invention provides a method for preparing the above-mentioned composition, which involves mixing glycyrrhizin and glutathione.

[0018] In this invention, the method of obtaining glycyrrhizin is not limited. It can be obtained directly through commercial purchase, or it can be prepared from Glycyrrhiza plants (Glycyrrhiza glabra, Glycyrrhiza uralensis, Glycyrrhiza inflata, Glycyrrhiza spinosa) through separation and extraction processes. It can also be prepared by chemical synthesis, biosynthesis (microbial fermentation synthesis, plant cell culture synthesis), etc. Regardless of the preparation process used, as long as the final active ingredient is glycyrrhizin, it falls within the scope of raw materials that can be used in this invention.

[0019] In this invention, the method of obtaining glutathione is not limited. It can be obtained directly through commercial purchase, or it can be prepared from animal tissues, microorganisms, and plants as extraction raw materials through extraction processes. It can also be prepared by chemical synthesis methods (liquid-phase synthesis, solid-phase synthesis), biosynthesis methods (microbial fermentation, enzymatic synthesis), etc. Regardless of the preparation process used, as long as the final active ingredient obtained is glutathione, it falls within the scope of raw materials that can be used in this invention.

[0020] Thirdly, the present invention provides the use of the above-described composition in the preparation of products for skin whitening or inhibiting melanin formation.

[0021] Specifically, the products include topical and oral formulations.

[0022] Furthermore, the topical preparations include serums, lotions, creams, masks, and lotions.

[0023] Furthermore, when the product is a topical preparation, it may include matrix raw materials and conventional excipients.

[0024] More specifically, the matrix raw materials include, but are not limited to, gelatinous raw materials, powdery raw materials, oily raw materials, waxy raw materials, coagulants, and surfactants.

[0025] More specifically, the aforementioned conventional excipients include, but are not limited to, antioxidants, adhesives, lubricants, humectants, preservatives, whitening agents, film-forming agents, emulsifiers, and cosmetic nutritional additives.

[0026] Specifically, the product is an oral formulation.

[0027] Specifically, the product may also include food-grade additives.

[0028] Furthermore, the acceptable excipients in the food are selected from one or more of acidity regulators, sweeteners, preservatives, antioxidants, stabilizers, emulsifiers, and leavening agents.

[0029] Furthermore, the oral formulations include tablets, capsules, powders, gel candies, oral liquids, or granules.

[0030] Fourthly, the present invention provides an oral whitening product comprising the above-described composition.

[0031] Specifically, the product may also include food-grade additives.

[0032] Furthermore, the acceptable excipients in the food are selected from one or more of acidity regulators, sweeteners, preservatives, antioxidants, stabilizers, emulsifiers, and leavening agents.

[0033] Furthermore, the acidity regulator is selected from one or more of citric acid, malic acid, lactic acid, tartaric acid, and acetic acid;

[0034] Furthermore, the sweetener is selected from one or more of xylitol, erythritol, sorbitol, maltitol, steviol glycosides, honey, and sucralose; Furthermore, the preservative is selected from one or more of potassium sorbate, sodium benzoate, sodium dehydroacetate, calcium propionate, and nitrite; Furthermore, the antioxidant is selected from one or more of vitamin E, ascorbic acid, and tea polyphenols; Furthermore, the stabilizer is selected from one or more of pectin, agar, gelatin, guar gum, xanthan gum, starch, hydroxypropyl starch, gum arabic, and hydroxypropyl methylcellulose; Furthermore, the emulsifier is selected from one or more of soybean lecithin, egg yolk lecithin, mono- and diglycerides of fatty acids, sucrose fatty acid esters, Span, polysorbate, and sodium caseinate; Furthermore, the leavening agent is selected from one or more of sodium bicarbonate, ammonium bicarbonate, ammonium carbonate, potassium aluminum sulfate, potassium hydrogen tartrate, and calcium hydrogen phosphate.

[0035] Furthermore, the oral formulations include tablets, capsules, powders, gel candies, oral liquids, or granules.

[0036] The beneficial effects of this invention are as follows: The two components in the whitening composition of this invention work synergistically to reduce melanin production and pigment content. Without affecting the normal vitality of skin cells, it achieves a powerful inhibition of melanin, providing a new and effective solution for the development of oral whitening products. Detailed Implementation

[0037] The present invention will be further described in detail below with reference to specific embodiments. The following embodiments are not intended to limit the present invention, but only to illustrate the present invention. Unless otherwise specified, the experimental methods used in the following embodiments are generally performed under conventional conditions. Unless otherwise specified, the materials and reagents used in the following embodiments are commercially available.

[0038] Source of experimental materials: The licorice root was ordered from MCE Company, product number HY-N0393.

[0039] Glutathione was purchased from MCE, catalog number HY-D0187.

[0040] Example 1 Preparation of the composition The composition consists of 1 part glycyrrhizin and 63 parts glutathione, with a mass ratio of glycyrrhizin to glutathione of 1:63.

[0041] The composition is prepared by mixing glycyrrhizin and glutathione.

[0042] Example 2 Preparation of the composition The composition consists of 1 part glycyrrhizin and 10 parts glutathione, with a mass ratio of glycyrrhizin to glutathione of 1:10.

[0043] The composition was prepared using the same method as in Example 1.

[0044] Example 3 Preparation of the composition The composition consists of 1 part glycyrrhizin and 200 parts glutathione, with a mass ratio of glycyrrhizin to glutathione of 1:200.

[0045] The composition was prepared using the same method as in Example 1.

[0046] Example 4 Preparation of the composition The composition consists of 1 part glycyrrhizin and 125 parts glutathione, with a mass ratio of glycyrrhizin to glutathione of 1:125.

[0047] The composition was prepared using the same method as in Example 1.

[0048] Example 5 Preparation of the composition The composition consists of 1 part glycyrrhizin and 1 part glutathione, with a mass ratio of 1:1 between glycyrrhizin and glutathione.

[0049] The composition was prepared using the same method as in Example 1.

[0050] Comparative Example 1 Compared with Example 1, the only difference is that Comparative Example 1 contains only 1 part of glycyrrhizin.

[0051] Comparative Example 2 The only difference from Example 1 is that Comparative Example 2 contains only 63 portions of glutathione.

[0052] Comparative Example 3 Compared with Example 1, the only difference is that the mass ratio of glycyrrhizin to glutathione is different, specifically 1:220. The composition in Comparative Example 3 includes 1 part glycyrrhizin and 220 parts glutathione.

[0053] Comparative Example 4 Compared with Example 1, the only difference is that the mass ratio of glycyrrhizin to glutathione is different, specifically 1:0.5. The composition in Comparative Example 4 includes 1 part glycyrrhizin and 0.5 parts glutathione.

[0054] Experiment Example 1: Cell Experiment Modeling method: Mouse B16-F10 melanoma cells, stimulation: α-MSH (100nM) to promote melanin production.

[0055] Experimental drug administration: The compositions obtained in Examples 1-5 and Comparative Examples 1-4 were dissolved in physiological saline: In Example 1, the concentration of glycyrrhizin was 3 ug / ml, and the concentration of glutathione was 189 ug / ml; In Example 2, the concentration of glycyrrhizin was 3 ug / ml, and the concentration of glutathione was 30 ug / ml; In Example 3, the concentration of glycyrrhizin was 3 ug / ml, and the concentration of glutathione was 600 ug / ml; In Example 4, the concentration of glycyrrhizin was 3 ug / ml, and the concentration of glutathione was 375 ug / ml; In Example 5, the concentration of glycyrrhizin was 3 μg / ml, and the concentration of glutathione was 3 μg / ml; In Comparative Example 1, the concentration of glycyrrhizin was 3 μg / ml; The concentration of glutathione in Comparative Example 2 was 189 ug / ml; In Comparative Example 3, the concentration of glycyrrhizin was 3 ug / ml, and the concentration of glutathione was 660 ug / ml. In Comparative Example 4, the concentration of glycyrrhizin was 3 ug / ml, and the concentration of glutathione was 1.5 ug / ml.

[0056] Testing indicators: 1. CCK8 cell viability (IC50, ELISA reader assay) Experimental methods: Cells were seeded at 1000 cells / well in 96-well plates and treated with 10 μM of the enzymes from Examples 1-5 and Comparative Examples 1-4 for 48 h. After incubation in CCK8 for 2 h, the cells were detected using a microplate reader. Changes in cell viability under different concentrations of different raw materials were calculated using OD values.

[0057] The cell viability of the control group was 100%, and the cell viability value was calculated as: experimental group / control group.

[0058] Experimental results: The results are shown in Table 1: Table 1

[0059] 2. Melanin content (statistics after photographing with Masson-Fontana melanin staining) Experimental method: Cells were packed at 2 x 10⁻⁶ cm⁻¹. 5 The samples were laid in 6-well plates. The components of Examples 1-5 and Comparative Examples 1-4 were treated for 48 hours. Each group had 3 replicate wells. The samples were stained with Masson-Fontana melanin (Solepro G2032, operated according to the instructions). After taking pictures under a microscope, the proportion of melanin area to the total area was counted.

[0060] Experimental results: Table 2

[0061] 3. Tyrosinase activity (detected using kit and microplate reader) Experimental method: Cells were packed at 2 x 10⁻⁶ cm⁻¹. 5The samples were seeded in 6-well plates. The components of Examples 1-5 and Comparative Examples 1-4 were treated for 48 hours. Each group had 3 replicate wells. The effects of different combinations on cellular tyrosinase activity were detected according to the steps of the tyrosinase activity kit (Solepro, BC4055).

[0062] Table 3 Tyrosinase Inhibition Rate

[0063] Experiment Example 2: Zebrafish Experiment I. Experimental Method for Detecting Melanin Production Inhibition Rate: 1 Sample Preparation Negative control group: Holt-Buffer culture medium was used as the negative control group; Example 1 group: The composition of Example 1 was diluted with Holt-Buffer culture medium to make the concentration of glycyrrhizin 0.36 mg / L and the concentration of glutathione 22.68 mg / L; Example 2 group: The composition of Example 2 was diluted with Holt-Buffer culture medium so that the concentration of glycyrrhizin was 0.36 mg / L and the concentration of glutathione was 3.6 mg / L. Example 3 group: The composition of Example 3 was diluted with Holt-Buffer culture medium so that the concentration of glycyrrhizin was 0.36 mg / L and the concentration of glutathione was 72 mg / L. Example 4 group: The composition of Example 4 was diluted with Holt-Buffer culture medium so that the concentration of glycyrrhizin was 0.36 mg / L and the concentration of glutathione was 45 mg / L. Example 5 group: The composition of Example 5 was diluted with Holt-Buffer culture medium to make the concentration of glycyrrhizin 0.36 mg / L and the concentration of glutathione 0.36 mg / L; Comparative Example 1: The glycyrrhizin in Comparative Example 1 was diluted with Holt-Buffer culture medium to make the concentration of glycyrrhizin 0.36 mg / L. Comparative Example 2: The glutathione in Comparative Example 2 was diluted with Holt-Buffer culture medium to make the concentration of glutathione 22.68 mg / L; Comparative Example 3: The composition of Comparative Example 3 was diluted with Holt-Buffer culture medium so that the concentration of glycyrrhizin was 0.36 mg / L and the concentration of glutathione was 79.2 mg / L. Comparative Example 4: The compositions of Comparative Example 4 were diluted with Holt-Buffer culture medium so that the concentration of glycyrrhizin was 0.36 mg / L and the concentration of glutathione was 0.18 mg / L.

[0064] 2. Model construction and drug administration: Healthy zebrafish embryos (6-8 hpf) were selected and placed in 96-well plates, six embryos per well. 200 μL of the above-mentioned sample was added to each well, with three replicates per group. The plates were incubated at 28.5 ± 0.5 °C. After 48 h of incubation, the pigment deposition of the embryos was observed under a microscope and photographed. The images were analyzed for grayscale values ​​using ImageJ software to calculate the relative melanin content.

[0065] Relative melanin content = (Melanin content of control group / Melanin content of experimental group) × 100%; Melanin content refers to the total area containing melanin in grayscale analysis.

[0066] Formula for calculating the inhibition rate of melanin production in zebrafish embryos: Z% = (Z C -Z X ) / Z C ×100%; In the formula, Z represents the inhibition rate of melanin production in zebrafish embryos. X Z represents the relative melanin content of the sample group after the zebrafish experiment. C This indicates the relative melanin content in the negative group after zebrafish testing.

[0067] 3. Experimental Results As shown in the table below: Table 4. Inhibition rate of melanin production in zebrafish embryos

[0068] II. Tyrosinase Activity Detection 1. Experimental Methods: Healthy zebrafish embryos at 9 hpf post-fertilization were randomly selected and placed in 6-well cell culture plates, 30 embryos per well, with 200 μL of the above sample solution added to each well. Each group was divided into 3 replicates and incubated in a biochemical incubator.

[0069] After 72 hours of incubation, lysis buffer was added, and the mixture was homogenized by sonication. The homogenate was centrifuged, and the supernatant was transferred to a 96-well plate. Levodopa solution was added simultaneously, and the plate was incubated for 1 hour. The absorbance (OD475nm) of each experimental group was measured using a microplate reader. The tyrosinase activity inhibition rate was calculated using the following formula: Inhibition rate = 1 -

[0070] 2. Experimental Results Table 5. Tyrosinase activity inhibition rate

[0071] Experiment Example 3: Animal Experiment 1 Experimental Methods Seventy-two healthy male brown-yellow guinea pigs, 8-10 weeks old and weighing 240-280g, were acclimatized for one week and then divided into groups of six. The experimental groups are as follows: Model control group (oral administration): Administered an equal volume of distilled water by gavage; Example 1 Oral group: The composition prepared in Example 1 was dissolved in distilled water. When administering the drug, the dosage of glycyrrhizin was 0.11 mg / kg / day and the dosage of glutathione was 6.93 mg / kg / day; the oral gavage dose was 5 ml / kg. Comparative Example 1 Oral Group: The glycyrrhizin of Comparative Example 1 was dissolved in distilled water. The dosage of glycyrrhizin was 0.11 mg / kg / day; the oral gavage dose was 5 ml / kg. Comparative Example 2 Oral Group: Glutathione from Comparative Example 2 was dissolved in distilled water. The dosage of glutathione was 6.93 mg / kg / day; the gavage dose was 5 ml / kg. Comparative Example 3 Oral Group: The composition prepared in Comparative Example 3 was dissolved in distilled water. During administration, the dosage of glycyrrhizin was 0.11 mg / kg / day, and the dosage of glutathione was 24.2 mg / kg / day; the gavage dose was 5 ml / kg. Comparative Example 4 Oral Group: The composition prepared in Comparative Example 4 was dissolved in distilled water. When administering the drug, the dosage of glycyrrhizin was 0.11 mg / kg / day and the dosage of glutathione was 0.055 mg / kg / day; the oral gavage dose was 5 ml / kg.

[0072] Take the oral whitening extract for 30 consecutive days. First, use scissors to trim the long hair on the guinea pig's back, covering an area of ​​4cm. 2 ×4cm 2 After shaving the hair short, the guinea pigs were irradiated with UVB lamps on the hairless area of ​​their backs at 3, 5, 7, and 9 days after administration of the test substance. The lamps were 15 cm away from the backs, with a UV wavelength of 280-320 nm and a total irradiation dose of 200 mJ / cm² per session. 2 A total of 4 times were performed. The L value was measured using a colorimeter before irradiation (L1) and 30 days after medication (L2).

[0073] L = L2 - L1.

[0074] Model control group: propylene glycol: ethanol: water = 5:3:2 (v / v) solvent; applied twice daily; application amount the same as in Example 1 group; Example 1 Topical application group: A solution was prepared using propylene glycol, ethanol, and water. The daily dosage of glycyrrhizin was 0.11 mg / kg / day, and the daily dosage of glutathione was 6.93 mg / kg / day; the solution was applied twice daily. Comparative Example 1 Topical Application Group: A solution was prepared using propylene glycol, ethanol, and water. The daily dose of glycyrrhizin was 0.11 mg / kg / day; applied twice daily. Comparative Example 2: Topical application group: A solution was prepared using propylene glycol, ethanol, and water as solvents. The daily dose of glutathione was 6.93 mg / kg / day; applied twice daily. Comparative Example 3: Topical application group: A solution was prepared using propylene glycol, ethanol, and water. The daily dosage of glycyrrhizin was 0.11 mg / kg / day, and the daily dosage of glutathione was 24.2 mg / kg / day; the solution was applied twice daily. Comparative Example 4 (Topical Application Group): A solution was prepared using propylene glycol, ethanol, and water. The daily dosage of glycyrrhizin was 0.11 mg / kg / day, and the daily dosage of glutathione was 0.055 mg / kg / day; the solution was applied twice daily. Administration continued for 30 days, and L values ​​were measured using a colorimeter before and after administration.

[0075] L = L after administration - L before administration.

[0076] 2. Experimental Results Table 6 L value

[0077] Note: Compared with the model control group (oral administration) p <0.01.

[0078] Table 7 L value

[0079] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An oral whitening efficacy composition, characterized in that, The composition consists of glabridin and glutathione.

2. The composition of claim 1, wherein, The mass ratio of glabridin and glutathione in the composition is 1:1-200.

3. The composition of claim 2, wherein, The mass ratio of glabridin and glutathione in the composition is 1:10-125.

4. The composition of claim 3, wherein, The mass ratio of glabridin and glutathione in the composition is 1:50-80.

5. Process for the preparation of a composition according to any one of claims 1 to 4, characterized in that, Glabridin and glutathione are mixed.

6. Use of the composition of any one of claims 1-4 in the preparation of a product for skin whitening or inhibiting melanin formation.

7. Use according to claim 6, characterized in that, The product further comprises a food acceptable adjuvant.

8. Use according to claim 7, characterized in that, The food acceptable adjuvant is selected from one or more of acidity regulator, sweetener, preservative, antioxidant, stabilizer, emulsifier, leavening agent.

9. Use according to claim 6, characterized in that, The product is in the form of oral preparation.

10. An oral whitening product, characterized in that, The oral whitening product comprises the composition of any one of claims 1-4.

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