Composition and dietary supplement containing same

Abstract

The invention discloses a composition which is prepared from the following components in parts by weight: 5 to 50 parts of ergothioneine, 2000 to 10000 parts of collagen peptide, 10 to 150 parts of sodium hyaluronate and 30 to 200 parts of aloe extract. The invention further discloses an anti-oxidation, anti-aging and whitening dietary supplement containing the composition and food acceptable auxiliary materials. Various components in the composition and the dietary supplement containing the composition are green and environment-friendly in source, safe, non-irritant and easy to be directly absorbed by intestinal tracts, the synergistic effect can improve the skin and health state from the deep level, and various requirements of oxidation resistance, aging resistance, inflammation resistance, whitening, filling and the like are met.

Description

Technical Field

[0001] The present invention belongs to the technical field of health food and dietary supplements, and in particular relates to a composition and a dietary supplement containing the same. Background Art

[0002] Ergothioneine is a natural rare chiral amino acid that comes from plants and can be accumulated in animals. It is safe and non-toxic. It is a natural antioxidant with multiple physiological functions such as detoxification, maintaining DNA growth, anti-radiation, anti-inflammation, anti-aging, protecting the nervous system, inhibiting developmental defects, protecting the liver, etc. It is very important to human health.

[0003] Collagen peptides are hydrolyzates of collagen, which are polypeptides or small peptide mixtures. In the food health care, biomedicine and cosmetics industries, collagen peptides have a wide range of applications and huge market value. Especially in cosmetics, since collagen peptides are similar in structure to human skin collagen and have good compatibility, they can penetrate deep into the human skin and have a good nutritional effect on the skin, and are widely used in the cosmetics industry.

[0004] Sodium hyaluronate, also known as sodium hyaluronate, is an important component of cell matrix and various tissues, and has many important physiological functions. Sodium hyaluronate has unique viscoelasticity, biocompatibility and degradability. It can improve the microcirculation of blood in the skin and nutrient absorption, helping to maintain normal metabolism. It has super moisturizing properties, can reduce wrinkles and make the skin smoother and softer; it can penetrate into the dermis, promote blood microcirculation, facilitate the skin's absorption of nutrients, and play an anti-wrinkle role.

[0005] Aloe vera extract has strong hydrating properties and is often used as a facial mask, which is very popular among women. Aloe vera's anthraquinone compounds have the properties of astringent, softening, moisturizing, anti-inflammatory, and tenderizing the skin, and also have the effects of relieving hardening, keratinization, and improving scars.

[0006] Affected by age, diet, stress, ultraviolet rays, etc., human skin will age and degenerate to varying degrees as external physical factors and internal physiological factors change. With the development of the economy and the improvement of living standards, people are more and more eager to pursue beauty, and continue to use cosmetics and skin care products to beautify and nourish their skin. However, in recent years, more and more people have realized that cosmetics and skin care products alone cannot solve all the problems of beauty. Only by conditioning from the inside can beauty and health from the inside out be fundamentally achieved. At present, compositions with antioxidant, anti-aging, whitening and anti-inflammatory effects are relatively rare on the market, and there are no oral beauty products using such compositions. Summary of the invention

[0007] In one aspect, the present invention provides a composition, which is composed of the following components by weight: 5-50 parts of ergothioneine, 2000-10000 parts of collagen peptides, 10-150 parts of sodium hyaluronate, and 30-200 parts of aloe extract. In some embodiments, the composition is composed of the following components by weight: 20-25 parts of ergothioneine, 5000-7000 parts of collagen peptides, 30-60 parts of sodium hyaluronate, and 80-100 parts of aloe extract.

[0008] In some embodiments, the composition is composed of the following components by weight: 10-30 parts of ergothioneine, 4000-8000 parts of collagen peptides, 20-100 parts of sodium hyaluronate, 50-100 parts of aloe extract, 80-160 parts of blueberry extract, 50-200 parts of vitamin C, 3-30 parts of vitamin E, and 3-20 parts of niacinamide. In some embodiments, the composition is composed of the following components by weight: 15-25 parts of ergothioneine, 4000-8000 parts of collagen peptides, 20-50 parts of sodium hyaluronate, 50-100 parts of aloe extract, 80-120 parts of blueberry extract, 60-120 parts of Vitamin C, 5-15 parts of vitamin E, 5-10 parts of niacinamide.

[0009] In some embodiments, the composition further comprises one or both of natural rosehip essence and natural grape seed essence. In some embodiments, the composition further comprises 50-2000 parts of natural rosehip essence and natural grape seed essence. In some embodiments, the composition further comprises 50-600 parts of natural rosehip essence and / or 300-1500 parts of natural grape seed essence.

[0010] In some embodiments, the collagen peptide is a small molecule collagen peptide of less than 1000 Daltons, and the sodium hyaluronate is a small molecule sodium hyaluronate.

[0011] In another aspect, the present invention provides an antioxidant, anti-aging and whitening dietary supplement comprising the composition as described above and a food-acceptable auxiliary material.

[0012] In some embodiments, excipients include one or more of solvents, flavoring agents, colorants, excipients, and other food additives.

[0013] In some embodiments, the dietary supplement is prepared as an oral liquid, tablet, capsule, pill, granule, or powder.

[0014] In some embodiments, the dietary supplement is prepared as an oral liquid.

[0015] In another aspect, the present invention discloses a method for preparing the dietary supplement as described above, the method comprising: (1) mixing a formula amount of ergothioneine, collagen peptide, sodium hyaluronate, aloe extract, and optionally blueberry extract, vitamin C, vitamin E, and niacinamide to obtain a composition; (2) mixing the composition with auxiliary materials to obtain a dietary supplement.

[0016] In some embodiments, a composition is obtained by mixing a formulated amount of ergothioneine, collagen peptide, sodium hyaluronate, aloe vera extract, and optionally blueberry extract, vitamin C, vitamin E, niacinamide, and one or two of natural rosehip extract and natural grape seed extract.

[0017] The composition of the present invention and the dietary supplement containing the same have multiple components of green and environmentally friendly origin, safe and non-irritating ingredients, and are easily directly absorbed by the intestines. The multiple components work synergistically to improve the skin and health status from a deep level, and meet multiple needs such as anti-oxidation, anti-aging, anti-inflammatory and whitening. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The cytotoxicity of the composition at different doses is demonstrated.

[0019] Figure 2 The anti-inflammatory effect of the composition on UV-damaged cytokines was demonstrated as a result of the concentration of TNFa.

[0020] Figure 3 The anti-inflammatory effect of the composition on UV-damaged cytokines was demonstrated, as a result of the concentration of IL-8.

[0021] Figure 4 The anti-inflammatory effect of the composition on UV-damaged cytokines was demonstrated, as a result of the concentration of IL-6.

[0022] Figure 5 Glycosylation inhibition rates are shown.

[0023] Figure 6 The effects of the composition on UV-induced mtDNA co-deletion (Cdel) in fibroblasts are demonstrated.

[0024] Figure 7 A comparison of the skin conditions of the subjects after using the composition for 12 weeks is shown. DETAILED DESCRIPTION

[0025] Now will refer to preferred embodiments of the present invention in detail, further illustrate its embodiment.Although the present invention will be described in conjunction with preferred embodiments, it should be understood that they are not intended to limit the present invention to these embodiments.On the contrary, the present invention is intended to cover substitutions, modifications and equivalents, which may be included within the spirit and scope of the present invention as defined in the claims.

[0026] As used herein, the term "or" is intended to include "and" and "or." In other words, the term "or" may also be replaced with "and / or."

[0027] As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0028] As used herein, the term "comprises" or "includes" or variations thereof refer to the following instances where the term is used in its non-limiting sense, meaning that items following the term are included, but items not specifically mentioned are not excluded. It also includes the more restrictive verbs 'consisting essentially of' and 'consisting of'.

[0029] The technical features, implementation methods and beneficial effects of the present invention are further described in detail below in conjunction with specific implementation cases. The embodiments described below are only part of the embodiments of the present invention, not all of the embodiments. Unless otherwise specified, the materials and reagents described in the following embodiments are all common commercial products and can be purchased on the market. Example 1

[0030] The composition of Example 1 is prepared by mixing the following components: 25 parts of ergothioneine, 7000 parts of collagen peptides, 40 parts of sodium hyaluronate, 80 parts of aloe vera extract, and 500 parts of natural rosehip essence. Example 2

[0031] The composition of Example 2 is prepared by mixing the following components: 20 parts of ergothioneine, 5000 parts of collagen peptides, 50 parts of sodium hyaluronate, 100 parts of aloe vera extract, 100 parts of natural rosehip essence, and 800 parts of natural grape seed essence. Example 3

[0032] The composition of Example 3 is prepared by mixing the following components: 20 parts of ergothioneine, 4000 parts of collagen peptide, 20 parts of sodium hyaluronate, 120 parts of aloe vera extract, 100 parts of blueberry extract, 80 parts of vitamin C, 10 parts of vitamin E, 8 parts of niacinamide, 1200 parts of natural grape seed extract Example 4

[0033] The composition of Example 4 is prepared by mixing the following components: 15 parts of ergothioneine, 6000 parts of collagen peptides, 30 parts of sodium hyaluronate, 50 parts of aloe vera extract, 80 parts of blueberry extract, 100 parts of vitamin C, 5 parts of vitamin E, 5 parts of niacinamide, and 300 parts of natural rosehip essence. Example 5 Antioxidant Test

[0034] Group A is a solution with a concentration of 1 mg / mL made from the composition of Example 1, and Group B is 1 mg / mL of CoQ10, and the antioxidant capacity is tested.

[0035] Vitamin ETrolox water-soluble analogues were used as quantitative standards and analyzed by a fluorescence microplate analyzer. The presence of antioxidants can inhibit the fluorescence changes caused by free radicals, and the degree of inhibition reflects its antioxidant capacity against free radicals; the higher the oxygen radical absorption capacity (ORAC), the stronger the ability to inhibit free radicals. As shown in Table 1, the ORAC of the composition of Example 1 is much higher than that of other antioxidants such as CoQ10, indicating that Group A has strong antioxidant activity. Table 1 ORAC(umol Trolox / g) Group A 134.92 Group B 8.93 Example 6 Anti-inflammatory Test

[0036] The human normal human epidermal keratinocyte (NHEK) cell line HaCat was maintained at 95% air and 5% carbon dioxide at 37°C and cultured in DMEM supplemented with 10% heat-inactivated fetal bovine serum FBS, 2mM glutamine, and 1% penicillin / streptomycin. The culture was harvested and the cell morphology was examined using a phase contrast microscope; the cell suspension was counted using a hemocytometer to monitor the cell number.

[0037] The cells were seeded in a 96-well plate and treated with the composition of Example 3 at concentrations of 1 μM, 10 μM, 50 μM, and 100 μM for 72 h, and incubated with MTT solution (100 μg / well) for 4 h. The formazine precipitate was dissolved in dimethyl sulfoxide, and the absorbance at 570 nm was measured using an ELISA reader to quantitatively determine the MTT reduction degree. Figure 1 As shown, the cell activities at several dosages were all around 100% and not less than 85%; the composition of Example 3 at a concentration of 100 μM was non-toxic to the cells.

[0038] Cells were seeded in 12-well plates at a density of 50,000 cells / mL and cultured to approximately 80% confluency. Cells were washed once with phosphate-buffered saline (PBS) at room temperature, PBS was added to the cells, and then exposed to 50 mJ / cm 2 After irradiation, PBS was immediately replaced with culture medium containing the composition of Example 3 (the highest non-cytotoxic dose was 100 μM) and control (DMSO), and the cells were returned to 37° C. for 24 h.

[0039] Detect TNFa, IL-8, and IL-6 in the cell culture supernatant, and use the reference cytokine samples provided in the kit to generate a standard curve. The cell culture supernatant is pre-coated with the antibody at the bottom of the microplate, incubated and thoroughly washed. The substrate TMB is used for color development. TMB turns blue under the catalysis of peroxidase and turns into the final yellow under the action of acid; the color depth is positively correlated with the inflammatory factors in the sample. Use an enzyme reader to measure the absorbance (OD value) at a wavelength of 450nm, and calculate the sample concentration based on the standard curve. Figure 2-4 As shown, the composition of Example 3 can inhibit the production of inflammatory factors TNFa, IL-8, and IL-6, and there are significant differences compared with the control group, so it has a good anti-inflammatory effect on ultraviolet damage to cells. Example 7 AGEs inhibition test

[0040] A bovine serum albumin solution with a concentration of 1 mg / mL was dissolved in PBS and incubated with a concentration of 10 mM D-ribose and the composition of Example 2 at 37°C for 10 days, and the formation of glycosylation products was observed in the absence (Group A) and presence (Group B) of the composition of Example 2.

[0041] It is known that the final glycation end products emit light in the range of 420-600nm after excitation with light of wavelength 300-420nm. Glycation is assessed by measuring the fluorescence of AGEs at λem=450nm after the sample is excited at λex=380nm. The inhibition of glycation is determined by the reduction of fluorescence compared to samples treated with sugar alone. Figure 5 As shown, group B showed a higher glycosylation inhibition rate (about 120%). Example 8 Cell activity test

[0042] Normal human dermal fibroblasts (NHDF) were seeded in 6-well plates and cultured in DMEM basal medium supplemented with 10% FBS for 24 h. The medium was replaced with medium containing or without the composition of Example 4, tested at 20 μM, and the cells were pre-incubated for 24 h. After pre-incubation (D0), the medium was removed and the cells were placed in irradiated medium (without the composition of Example 4 or control) and UVA 4 J / cm 2Irradiation. After the irradiation, the culture medium was replaced (irradiation control group) and incubated for 3 hours. Repeat 3 times a day, and continue treatment after each irradiation. After 16 days of treatment, irradiation was performed on D1, D2, D6, D7, D8, D9, D13 and D15, irradiation was performed 3 times a day, and treatment was continued after each irradiation. On the 16th day, only 2 irradiation and treatment schemes were performed. At D5, the culture medium was replaced by culture medium containing or not containing Example 4 or the reference product (irradiation control). On the 12th day, the first treatment was performed in the morning and the second treatment was performed in the evening. All experimental conditions were performed in n=6 (at D16, n=3 for mtDNA depletion analysis and n=3 for activity analysis). At the end of incubation, the cells used for mtDNA depletion analysis were washed with PBS and frozen at -80°C.

[0043] Total genomic DNA (gDNA) was extracted from each experimental condition (duplicate samples were pooled before extraction) using a Nucleocapsid tissue kit, and gDNA content was measured using a spectrophotometer.

[0044] Amplification of expected fragment: A pair of primers was used to amplify the 262 bp of the retained region and relative quantification of total mtDNA. A pair of primers located on both sides of the deletion region (primers B and A) was used for relative quantification of Cdel, resulting in a 522 bp fragment amplification (the time of PCR extension was selected to avoid amplification of the non-deleted region; more than 5000 bp). The reaction mixture (10 μl of final product) was prepared as follows: - 2.5 μl of gDNA, - primers (forward and reverse), reagent mix ( Fast qPCR premix; Ozyme). PCR products were subjected to 1.5% agarose gel electrophoresis and detected with ethidium bromide. The gel was photographed under UV transillumination and the images were semi-quantitatively analyzed using Multigauge software.

[0045] At the end of the incubation, the cells for activity assessment were incubated with WST-8 (a highly water-soluble tetrazolium salt) which is reduced by succinate dehydrogenase (a mitochondrial enzyme) to a water-soluble orange product (formazan). This conversion is proportional to the number of living cells and their metabolic activity. The absorbance (OD) value of the extract at 450nm was recorded using a spectrometer.

[0046] The raw data were analyzed using the following formula: standard error of the mean: sem = Sd / n; activity percentage: activity (%) = (OD 样品 / OD 对照 )x100.

[0047] Under this experimental condition, at concentrations of 0.5 μM, 3 μM and 30 μM, the composition of Example 4 can significantly reduce the incidence of Cdel in UVA-irradiated NHDF (63%, 32% and 23%, respectively), indicating that it has a strong protective effect against UVA-mediated mtDNA deletion. Table 2. Effects on fibroblast activity after 72 hours of incubation Example 9 Skin color test

[0048] The formula contains the moisturizer of the composition of Example 4 and the comparative composition, and the moisturizer further contains one or more ingredients selected from the following: hyaluronic acid (0.1-3%), retinol / retinoic acid (0.1-2%), hydrolyzed collagen / collagen peptide (0.1-3%), vitamin C (0.1-6%), vitamin E (0.1-10%), EGCG / green tea (0.1-10%), polyhydroxy acid (0.1-12%) , glycolic acid (0.1-10%), aloe vera (0.1-10%), niacinamide (0.1-10%), resveratrol (0.1-5%), superoxide dismutase (0.1-5%), tea tree oil (0.1-10%), bakuchiol (0.1-5%), grape seed oil (0.1-10%), petrolatum (0.1-5%) or squalene (0.1-10%) (w / w) or other suitable conventional moisturizer ingredients.

[0049] Thirty healthy female subjects with mild to moderate photoaging and Fitzpatrick skin types I-II and aged 35-70 years were recruited for the clinical efficacy study. Inclusion and exclusion criteria were considered.

[0050] After enrollment, subjects will be randomly assigned to use the composition of the present invention. Subjects will be given a diary to record the daily use of the moisturizer, provide a return schedule, and are required to return to the research center in week 12. Subjects will take photos of their skin status every day for record. Control group: The same experimental group subjects will be used as the control group, and the skin status will be photographed and archived one week before the experiment as an experimental comparison. At week 12 (the end of the study), the subject's diary will be checked for compliance and collected.

[0051] Figure 7 The comparison of skin conditions after the subjects used the composition for 12 weeks is shown. Figure 7 As shown, after 12 weeks, the subjects' melanin decreased by 21%, spots decreased by 13%, and skin radiance improved by 79%.

[0052] The composition of the present invention and the dietary supplement containing the same have multiple components of green and environmentally friendly origin, safe and non-irritating ingredients, and are easily directly absorbed by the intestines; wherein, by rationally selecting the components and their contents, the multiple components work synergistically, can deeply improve the skin and health status, have excellent antioxidant properties, and can also meet multiple needs such as anti-aging, anti-inflammatory whitening, and brightening skin tone; and the oral product has a pleasant appearance and smell.

[0053] The above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Any technician familiar with the present invention may make various changes, modifications, substitutions and variations to these embodiments without departing from the principles and purpose of the present invention. The scope of the present invention is defined by the claims and their equivalents.

Claims

1. A composition, characterized in that The invention is composed of the following components by weight: 5-50 parts of ergothioneine, 2000-10000 parts of collagen peptides, 10-150 parts of sodium hyaluronate and 30-200 parts of aloe extract.

2. The composition according to claim 1, characterized in that The composition consists of the following components by weight: 10-30 parts of ergothioneine, 4000-8000 parts of collagen peptides, 20-100 parts of sodium hyaluronate, 50-100 parts of aloe extract, 80-160 parts of blueberry extract, 50-200 parts of vitamin C, 3-30 parts of vitamin E, and 3-20 parts of niacinamide.

3. The composition according to claim 1 or 2, characterized in that The composition further contains one or both of natural rosehip essence and natural grape seed essence.

4. The composition according to claim 1 or 2, characterized in that The collagen peptide is a small molecule collagen peptide below 1000 Daltons, and the sodium hyaluronate is a small molecule sodium hyaluronate.

5. An antioxidant, anti-aging and whitening dietary supplement, characterized in that: The dietary supplement comprises the composition according to claim 1 or 2 and a food-acceptable auxiliary material.

6. The dietary supplement according to claim 5, characterized in that The auxiliary materials include one or more of solvents, flavoring agents, colorants, excipients and other food additives.

7. The dietary supplement according to claim 5 or 6, characterized in that The dietary supplement is prepared in the form of oral liquid, tablet, capsule, pill, granule or powder.

8. The dietary supplement according to claim 5 or 6, characterized in that The dietary supplement is prepared as an oral liquid.

9. A method for preparing a dietary supplement according to claim 8, characterized in that: The method comprises: (1) mixing the formulated amounts of ergothioneine, collagen peptide, sodium hyaluronate, aloe extract, and optionally blueberry extract, vitamin C, vitamin E, and niacinamide to obtain a composition; (2) mixing the composition with auxiliary materials to obtain a dietary supplement.

10. The preparation method according to claim 9, characterized in that: The composition is obtained by mixing a formula amount of ergothioneine, collagen peptide, sodium hyaluronate, aloe extract and optionally blueberry extract, vitamin C, vitamin E, niacinamide and one or two of natural rosehip essence and natural grape seed essence.

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