CN115025001B - Peptide composition with multiple whitening and freckle removing effects and application thereof - Google Patents
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- CN115025001B CN115025001B CN202210610039.XA CN202210610039A CN115025001B CN 115025001 B CN115025001 B CN 115025001B CN 202210610039 A CN202210610039 A CN 202210610039A CN 115025001 B CN115025001 B CN 115025001B
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/67—Vitamins
- A61K8/676—Ascorbic acid, i.e. vitamin C
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/36—Carboxylic acids; Salts or anhydrides thereof
- A61K8/365—Hydroxycarboxylic acids; Ketocarboxylic acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/494—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/494—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with more than one nitrogen as the only hetero atom
- A61K8/4946—Imidazoles or their condensed derivatives, e.g. benzimidazoles
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/67—Vitamins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/67—Vitamins
- A61K8/673—Vitamin B group
- A61K8/675—Vitamin B3 or vitamin B3 active, e.g. nicotinamide, nicotinic acid, nicotinyl aldehyde
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
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- Veterinary Medicine (AREA)
- Birds (AREA)
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- Emergency Medicine (AREA)
- Dermatology (AREA)
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a peptide composition with multiple whitening and freckle removing effects and application thereof, wherein the composition comprises 5-15 parts of snow carnosine, 2-7 parts of nonapeptide-1, 1-10 parts of carnosine, 3-8 parts of vitamin C, 1-5 parts of copper gluconate, 1-7 parts of tripeptide-1 and 2-8 parts of glutathione. The composition achieves the effects of whitening and removing spots from the cell level; the cosmetic can be applied to cosmetics, can solve the problems of damaged skin barrier, rough texture, uneven skin color, dark color, color spots and the like, is particularly suitable for dark skin, and can be used as an additive of cosmetics to be added into cosmetics such as toning lotion, cleansing cream, emulsion, essence, face cream, gel, facial mask and the like.
Description
Technical Field
The invention belongs to the technical field of cosmetics, and discloses a peptide composition with multiple whitening and freckle removing effects and application thereof.
Background
Is good for everyone. The beautiful appearance is a dream of every female, and the female has confidence. However, skin darkness, tanning, sunburn, freckle and chloasma seriously affect the mood of each love heart. The skin of a person is related to the race of the person, the basic skin tone being a result of natural and gradual synergy from the acquired. The color of the skin is related to the aggregation of melanin. The reasons for this are also various. In general, genetic factors, endocrine factors, ultraviolet and solar irradiation, radical action, air and environmental pollution, and the like are related.
The mechanism of melanin formation is as follows: the skin melanin formation process comprises a series of complex physiological and biochemical processes such as migration of melanocytes, division and maturation of melanocytes, formation of melanosomes, transportation of melanin particles, degradation of melanin and the like. From the biochemical reaction point of view, there are three basic substances necessary for melanin formation: tyrosine is the main raw material for preparing melanin; tyrosinase is the main rate limiting enzyme for converting tyrosine into melanin, and is a composition of copper and protein; tyrosine produces melanin under the action of tyrosinase, an oxidation process, which must be combined with oxygen to convert it to melanin. Any factors with the function of interrupting melanin formation have the functions of whitening and removing freckle. Second, skin aging is a significant cause of dark yellow complexion and pigmentation.
General whitening and freckle removing are divided into five dimensions: 1) Mainly from directly controlling and inhibiting various enzymes in the process of melanin cell generation; 2) Selectively destroying melanocytes, inhibiting the formation of melanosomes, and altering their structure; 3) Reducing dopamine; 4) Extracellular inhibition of melanocytes; 5) Exogenous influencing factors, such as accelerating cell renewal, etc.; 6) The cells are kept in a young state, and the cell viability is increased and the situation is vigorous.
From the whitening mechanism, almost all whitening products can change the self structure of the skin more or less, destroy melanocytes, inhibit melanosomes, change the environment of melanocytes, accelerate cell renewal and the like, destroy the protective barrier of the skin, and the trans-dermal water loss TEWL value is several times, even more than ten times that of the original skin. The skin is easy to get off, dry, even red and itchy, and the like, the skin barrier is damaged, and the influence of the environment, especially ultraviolet rays and sunlight, on the skin is increased, so that the problems of darker pigmentation and darkening skin color are possibly caused. In order to reduce the damage of the whitening and freckle removing agent to the self structure of the skin, the composition is generally used for increasing functional components such as repairing, relieving, moisturizing and the like so as to achieve the effect of protecting the skin. In fact, these methods are only exogenous methods, are repair methods, and are methods of repairing walls while removing the walls. If this balance is slightly disturbed, it is enhanced to whiten and remove spots, which may cause physiological disorders such as the Japanese "azalea" event, causing irreversible damage to the skin of hundreds of thousands of people. Otherwise, if the whitening and freckle removing effects are weakened, the expected effect cannot be achieved.
Therefore, a whitening and freckle-removing composition without changing skin functions is sought, and is an important point and difficulty in research and development in the fields of cosmetics and medicines.
Disclosure of Invention
The invention aims to provide a peptide composition with multiple whitening and freckle removing effects, which achieves the effects of whitening and freckle removing from the cellular level; the skin care product can be applied to cosmetics, can solve the problems of skin barrier repair, fine texture, uneven skin color, dark color, color spots and the like, and is particularly suitable for dark skin.
The peptide composition with multiple whitening and freckle removing effects provided by the invention comprises the following components in parts by weight: 5-15 parts of snow carnosine, 2-7 parts of nonapeptide-1, 1-10 parts of carnosine and 3-8 parts of vitamin C.
Further, the composition also comprises 1 to 5 parts of copper gluconate and 1 to 7 parts of tripeptide-1.
Further, the composition further comprises 2 to 8 parts of glutathione.
Preferably, the composition comprises the following components in parts by weight: 7-12 parts of snow carnosine, 3-6 parts of nonapeptide-1, 4-8 parts of carnosine and 4-7 parts of vitamin C.
More preferably, the composition comprises the following components in parts by weight: 8-11 parts of snow carnosine, 4-5 parts of nonapeptide-1, 5-7 parts of carnosine, 5-6 parts of vitamin C, 2-4 parts of copper gluconate and 3-6 parts of tripeptide-1.
More preferably, the composition comprises the following components in parts by weight: 8-11 parts of snow carnosine, 4-5 parts of nonapeptide-1, 5-7 parts of carnosine, 5-6 parts of vitamin C, 2-4 parts of copper gluconate, 3-6 parts of tripeptide-1 and 4-7 parts of glutathione.
The peptide composition with multiple whitening and freckle removing effects can be used as an additive of cosmetics to be added into cosmetics such as toning lotion, cleansing cream, emulsion, essence, face cream, gel, facial mask and the like. In production, the selection and preparation process of other components in the cosmetic can be performed according to conventional operations in the art.
Compared with the prior art, the invention has the following beneficial effects:
(1) The carnosine in the composition provided by the invention is a natural dipeptide, has the functions of resisting oxidation and free radicals, and is a main factor for resisting skin aging; the method can keep the cells in a young state, increase the activity of the cells and present vigorous situations; has good anti-glycation effect, can react with sugar, reduces the glycation reaction of gray yellow collagen, and avoids the brown yellow skin. Vitamin C is a naturally occurring antioxidant, and supplementation with vitamin C is an important route to avoid oxidative stress and skin aging, playing a vital role in collagen synthesis. Vitamin C is directly involved in melanin formation, and has melanin generation inhibiting effect. The chelation and antioxidation of carnosine can be mutually stable with the antioxidation of vitamin C, so that the oxidative degradation of vitamin C is reduced. The snow carnosine is composed of hydroxyethyl piperazine ethane sulfonic acid, AA2G and nicotinamide. The hydroxyethyl piperazine ethane sulfonic acid has the functions of updating epidermis, increasing transdermal absorption capacity, keeping the normal pH value of skin, absorbing ultraviolet rays, reducing the stimulation of the ultraviolet rays to melanocytes, reducing the formation of melanin in the skin, and having certain sun-screening and whitening effects. AA2G is a stable water-soluble vitamin C modifier. Nicotinamide is vitamin PP, which is itself able to penetrate the stratum corneum, increasing the transdermal absorption function of the active substance. The snow carnosine directly participates in physiological actions among cells, inhibits the transfer of melanosomes from melanocytes to cutin, and stops or delays the production of melanin by controlling the transfer path, so that the effects of whitening and removing freckles are achieved, and the skin yellowing is inhibited, and the snow carnosine has the synergistic effect with carnosine and vitamin C. The nonapeptide-1 can act on the first step of melanin formation, namely can compete with melanocyte-promoting hormone for melanocortin receptor, reduce the binding rate, block and inhibit the formation of melanin in the whole course, and has the functions of directly inhibiting the activity of melanocytes and reducing the synthesis of melanin by melanocytes. The snow carnosine can quickly penetrate into the skin, inhibit the generation of histamine, eliminate histamine, shrink vascular smooth muscle, increase the resting tension of aortic smooth muscle, eliminate the loss of sunlight and ultraviolet rays to the skin, inhibit the activity of the neuraminidase, accelerate the update of the epidermis of the skin and the like. The vitamin C is added to give sufficient nutrition to the skin, so that the skin metabolism is quickened while the repair is quickened, the lasting repair and moisturizing effects are achieved, moderate repair and moisturizing of the skin can be provided by cooperation with other components, the skin is conditioned to a balanced state, and therefore the skin is effectively whitened, freckle is removed and the barrier is restored.
(2) The copper gluconate in the composition provided by the invention is a copper salt with an alpha-hydroxy acid structure, has the effects of resisting oxidation and inflammation, can promote skin regeneration, reduce wrinkles and is beneficial to skin aging. Copper gluconate reacts with carnosine to generate carnosine copper which is a signal peptide with new efficacy, can be used as an activator and an antioxidant for tissue remodeling, and can promote degradation of a large amount of collagen aggregates outside scars, synthesis of normal collagen of skin, generation of elastin, proteoglycan and glycosaminoglycan, growth speed and migration of different cell types, anti-inflammatory and antioxidant reactions. Tripeptide-1 is a dermal growth factor, is a signal peptide, has excellent activation, activates a receptor, heals damaged or injured skin, can ensure a sufficient number of living keratin cells to participate in the reformation and migration of wound epidermis, and plays a very important physiological role in skin healing. Tripeptide-1 reacts with copper gluconate to form tripeptide-1 copper, which has repair and relief effects similar to carnosine copper. The snow carnosine, the nine peptide-1, the carnosine and the vitamin C are compounded, so that the effects of whitening skin, removing color spots, repairing barriers, nourishing, moisturizing and the like are achieved on the cellular level.
(3) The glutathione in the composition provided by the invention is composed of two forms of reduction type and oxidation type, and is tripeptide composed of glutamic acid, cysteine and glycine, and the excellent effects of the tripeptide are antioxidation, free radical removal, cell membrane lipid peroxidation inhibition, tyrosinase activity inhibition and the like. The glutathione and the vitamin C are in a stable and reinforced relation, and the combined use effect of the glutathione and the vitamin C is stronger than that of the single use in the aspects of inhibiting tyrosinase and resisting oxidization.
(4) The composition provided by the invention intervenes melanin synthesis from a molecular layer, such as inhibition of melanin by nonapeptide-1 and glutathione, intervention of melanin body delivery by nicotinamide, intervention of cell activity by carnosine, tripeptide-1 and copper gluconate and the like, are all completed under the condition of cell physiological functions, but the formula disclosed by the invention achieves the effects of whitening and freckle removing in an intangible way under the condition of not interfering with the cell physiological functions.
Detailed Description
In the following, it is apparent that the embodiments described are only some, but not all embodiments of the present invention by clearly and specifically describing the technical solutions of the present invention. For process parameters or conditions not specifically noted, reference may be made to conventional techniques.
The raw materials used in the invention are purchased through commercial sources. Wherein, vitamin C and copper gluconate are purchased from Guangzhou gamma energy biological limited company, carnosine and glutathione are purchased from Guangzhou Xeno biological technology limited company, and the snow carnosine, the nonapeptide-1 and the tripeptide-1 are purchased from Shanghai Rui trade limited company.
Example 1
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 0.5g of snow carnosine, 0.2g of nonapeptide-1, 0.1g of carnosine and 0.3g of vitamin C.
Example 2
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 0.7g of snow carnosine, 0.3g of nonapeptide-1, 0.4g of carnosine and 0.4g of vitamin C.
Example 3
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 0.9g of snow carnosine, 0.5g of nonapeptide-1, 0.6g of carnosine and 0.6g of vitamin C.
Example 4
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 1.2g of snow carnosine, 0.6g of nonapeptide-1, 0.8g of carnosine and 0.7g of vitamin C.
Example 5
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 1.5g of snow carnosine, 0.7g of nonapeptide-1, 1g of carnosine and 0.8g of vitamin C.
Example 6
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 0.5g of snow carnosine, 0.2g of nonapeptide-1, 0.1g of carnosine, 0.3g of vitamin C, 0.1g of copper gluconate 1 part and 0.1g of tripeptide-1.
Example 7
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 0.8g of snow carnosine, 0.4g of nonapeptide-1, 0.5g of carnosine, 0.5g of vitamin C, 0.2g of copper gluconate 1 part and 0.3g of tripeptide-1.
Example 8
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 1.1g of snow carnosine, 0.5g of nonapeptide-1, 0.7g of carnosine, 0.6g of vitamin C, 0.4g of copper gluconate 1 part and 0.6g of tripeptide-1.
Example 9
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 1.5g of snow carnosine, 0.7g of nonapeptide-1, 1g of carnosine, 0.8g of vitamin C, 0.5g of copper gluconate 1 part and 0.7g of tripeptide-1.
Example 10
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 0.5g of snow carnosine, 0.2g of nonapeptide-1, 0.1g of carnosine, 0.3g of vitamin C and 0.2g of glutathione.
Example 11
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 1.1g of snow carnosine, 0.5g of nonapeptide-1, 0.7g of carnosine, 0.8g of vitamin C and 0.7g of glutathione.
Example 12
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 1.5g of snow carnosine, 0.7g of nonapeptide-1, 1g of carnosine, 0.8g of vitamin C and 0.8g of glutathione.
Example 13
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 0.5g of snow carnosine, 0.2g of nonapeptide-1, 0.1g of carnosine, 0.3g of vitamin C, 0.1g of copper gluconate, 0.1g of tripeptide-1 and 0.2g of glutathione.
Example 14
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 0.8g of snow carnosine, 0.4g of nonapeptide-1, 0.5g of carnosine, 0.5g of vitamin C, 0.2g of copper gluconate, 0.3g of tripeptide-1 and 0.4g of glutathione.
Example 15
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 1.1g of snow carnosine, 0.5g of nonapeptide-1, 0.5g of carnosine, 0.6g of vitamin C, 0.4g of copper gluconate, 0.6g of tripeptide-1 and 0.7g of glutathione.
Example 16
The peptide composition with multiple whitening and freckle removing effects comprises the following components: 1.5g of snow carnosine, 0.7g of nonapeptide-1, 1g of carnosine, 0.8g of vitamin C, 0.5g of copper gluconate, 0.7g of tripeptide-1 and 0.8g of glutathione.
The preparation method of the compositions of the above examples 1 to 16 comprises: weighing the components, and stirring and mixing uniformly at normal temperature.
The use method of any one of the compositions in examples 1 to 16 is to add any one of the compositions to cosmetics, wherein the composition accounts for 0.5 to 8 percent of the total mass of the cosmetics, and skin care cosmetics such as toning lotion, cleansing cream, emulsion, essence, face cream, gel, mask and the like can be prepared according to the conventional process and auxiliary materials for preparing the cosmetics. In production, the selection and preparation process of other components in the cosmetic can be performed according to conventional operations in the art.
In order to demonstrate the effect of the present invention, comparative examples of the present invention are given below:
comparative example 1
The peptide composition with multiple whitening and freckle removing effects in the comparative example comprises the following components: 0.8g of carnosine and 0.5g of carnosine.
Comparative example 2
The peptide composition with multiple whitening and freckle removing effects in the comparative example comprises the following components: 0.4g of nonapeptide-1 and 0.5g of vitamin C.
Comparative example 3
The peptide composition with multiple whitening and freckle removing effects in the comparative example comprises the following components: 0.8g of carnosine, 0.5g of carnosine and 0.2g of copper gluconate.
Comparative example 4
The peptide composition with multiple whitening and freckle removing effects in the comparative example comprises the following components: 0.4g of nonapeptide-1, 0.5g of vitamin C and 0.3g of tripeptide-1.
Comparative example 5
The peptide composition with multiple whitening and freckle removing effects in the comparative example comprises the following components: 0.8g of snow carnosine, 0.5g of carnosine, 0.2g of copper gluconate and 0.4g of glutathione.
Comparative example 6
The peptide composition with multiple whitening and freckle removing effects in the comparative example comprises the following components: 0.4g of nonapeptide-1, 0.5g of vitamin C, 0.3g of tripeptide-1 and 0.4g of glutathione.
In order to better illustrate the advantages of the present invention, the following provides experimental effects of the technical solution provided by the present invention:
1. formula stability
Since vitamin C and glutathione are unstable and oxidized very easily, the stability of the compositions of examples 1 to 16 and comparative examples 1 to 6 was studied. The specific method comprises the following steps:
(1) Sample treatment: the compositions of examples 1 to 16 and comparative examples 1 to 6, which were just prepared, were designated as A 0 The method comprises the steps of carrying out a first treatment on the surface of the The compositions of examples 1 to 16 and comparative examples 1 to 6 were placed in an oven at 50℃for 30 days in the absence of light, and the data obtained were designated A 50 . The compositions of examples 1 to 16 and comparative examples 1 to 6 were each prepared as an aqueous solution having a mass concentration of 5%.
(2) The testing method comprises the following steps: vitamin C was tested by GB14754 method; glutathione is tested by glutathione test standards in Chinese pharmacopoeia.
(3) And (3) data processing: rate of change = 1-a 0 /A 50 The results are shown in Table 1.
TABLE 1 sample rate of vitamin C and glutathione change
As can be seen from Table 1, the stability of vitamin C was divided into four equal fractions, examples 1-5, examples 6-9, examples 10-12 and examples 13-16, respectively, from low to high. The result is due to the different ingredients of the formulation. The ingredients in each equivalent time are compared separately, and it can be seen that the proportion of the raw material components in the formula has little influence on the stability of the vitamin C, and the stability of the vitamin C is basically only related to the types of the raw materials. The same is true for the glutathione stability of examples 10 to 16. In comparative examples 2, 4 and 6, a large proportion of vitamin C was oxidized, decomposed and denatured due to the raw material components. The same is true for the glutathione in comparative example 5 and comparative example 6. In particular, comparative example 6 contains vitamin C and glutathione at the same time, but the decrease ratio is very large. The results of examples 1-16 are combined to demonstrate that the components of the compositions of the present invention are synergistic.
2. Whitening and freckle removing efficacy test
(1) Test sample: the compositions of examples 1 to 16 and comparative examples 1 to 6 were added to a general emulsion external matrix, respectively, to prepare test samples, the compositions accounting for 5% of the total mass of the emulsion; the control was 2% hydroquinone ointment (prescription drug, manufactured by Guangdong Kangdong pharmaceutical Co., ltd.) and the common emulsion topical matrix was used as a blank.
(2) The testing method comprises the following steps: b-16 melanoma cells were incubated with 0.1% glucosamine until complete albinism, and then 2mmol/L theophylline was added to restore melanin synthesis. And simultaneously adding a test sample, and microscopic examination of the color tone of the cell particles to judge the inhibition/promotion effect of the sample on the new melanin. And finally, centrifugally separating the cell particles to release the intracellular particles, measuring the absorption luminosity at 420nm, and measuring the total melanin amount to judge the influence of the test sample on the melanin amount.
Inhibition rate 100% = 100% of sample absorbance with intervention/absorbance without intervention of sample 100%
The greater the inhibition, the better the test sample inhibited melanin production. The results are shown in Table 2.
TABLE 2 test of sample inhibition of melanin
From the melanin test results of table 2, it can be seen that the expected effect was achieved in both examples and comparative examples, and the inhibition rate of melanocytes exceeded 40%. According to the different formula components, the melanin inhibition rate is superior to that of a control group, and the examples and the comparative examples have the effects of whitening and removing freckles. The melanin inhibiting effect was divided into four levels, examples 1 to 5, examples 6 to 9, examples 10 to 12 and examples 13 to 16, respectively, and the melanin inhibiting rate was from low to high, which was caused by the difference in the components of the test samples. The melanin inhibiting rate of comparative examples 1 to 6 was also superior to that of hydroquinone, and comparative examples 2, 4, 5 and 6 were inferior in stability test, and could not be used in conventional cosmetic formulations, and had feasibility in a secondary polishing product.
3. Human body safety inspection method
Human body safety test method the skin closed patch test is tested by referring to the human body skin patch test method of chapter 2 of cosmetic safety technical Specification (2015 edition).
(1) Test sample: the compositions of examples 1 to 16 and comparative examples 1 to 6 were each added to a general emulsion external matrix to prepare test samples, wherein the compositions account for 5% of the total mass of the emulsion. Meanwhile, 2% hydroquinone ointment is used as a control group, and a common emulsion external matrix is used as a blank group.
(2) The tester chooses: volunteers 18 to 60 years old, which meet the test requirements, were selected as subjects. The population with the following conditions could not be treated as a subject: antihistamines used in the last week or immunosuppressants used in the last month; any anti-inflammatory agent is applied to the tested part in the last two months; a subject suffering from a clinically unhealed inflammatory skin condition; insulin dependent diabetes mellitus patients; asthma or other chronic respiratory disease patients undergoing treatment; receiving an anticancer chemotherapeutic within approximately 6 months; patients with immunodeficiency or autoimmune disease; women in lactation or gestation; bilateral mastectomy and bilateral axillary lymphadenectomy; judging that the skin to be tested is affected by scars, pigments, atrophy, moles, or other flaws; participate in other clinical trial researchers; highly sensitive body constitution; non-volunteer participants or those who were unable to complete the prescribed content as required by the trial.
(3) The testing method comprises the following steps: 50 subjects were selected, 30 women, 20 men. The selected area is not more than 50mm 2 A plaque test apparatus of about 1mm depth. The test sample was placed in a plaque laboratory at a level of about 0.020 to 0.025g. The plaque test with the test sample applied to the forearm of the subject on the curved side with hypoallergenic tape and gently pressed with the palm to apply it uniformly to the skin for 24 hours. Skin reactions were observed according to the criteria of table 3 at 30 minutes (after the disappearance of the indentation), 24 hours and 48 hours after removal of the plaque tester of the test sample, respectively, and the observation results were recorded, and the results are shown in table 4.
TABLE 3 skin response grading Standard for skin seal Patch test
Table 4 patch test score
As can be seen from the patch results in Table 4, the compositions of examples 1 to 16 achieved the expected effects, all of which were negative reactions. However, comparative examples 1 to 4 have a great potential safety hazard, and comparative example 2 and comparative example 4 cause too strong irritation, and the test is stopped in advance, which is caused by the different components and proportions of comparative examples 1 to 6. As can be seen from the combination of the stability test, the efficacy test and the safety test results, although the melanin inhibition rates of comparative examples 1 to 4 are good, the safety of human bodies is hidden, and if the whitening agent is used, the whitening agent needs to be further perfected. Comparative examples 5 to 6 are viable for secondary polishing products.
4. Repair efficacy test
Reference T/ZHCA 003-2018 cosmetics were tested for their effect on the transepidermal water loss test method.
(1) Test sample: the compositions of examples 1 to 16 and comparative examples 1 to 6 were prepared as 5% aqueous solutions, respectively. Meanwhile, water is used as a control group.
(2) Subject selection
Number of subjects: the effective subjects were 220 persons, 10 persons per group.
Subject conditions: volunteers who met the trial at 18-60 years of age. The following cannot be selected as the subject: antihistamines used in the last week or immunosuppressants used in the last month; any anti-inflammatory agent is applied to the tested part in the last two months; a subject suffering from a clinically incurable inflammatory dermatological condition; insulin dependent diabetes mellitus patients; asthma or other chronic respiratory disease patients undergoing treatment; patients who received anti-cancer treatments in nearly six months; patients with immunodeficiency or autoimmune diseases; women in lactation or gestation; bilateral mastectomy and bilateral axillary lymphadenectomy; judging that the skin to be tested is affected by scars, pigments, atrophy, moles, or other blemishes; participate in other clinical trial researchers; highly sensitive body constitution; non-volunteer participants or those who were unable to complete the prescribed content as required by the trial.
(3) Measurement procedure
The measuring method comprises the following steps: the left and right forearm sides of the subject were randomly divided into a sample application side and a control side, ensuring that the sample application side and the control side were statistically balanced. The test parts were cleaned with clear water and blotted dry with a chipless blotter dry paper towel prior to each measurement. Sitting still for at least 20 minutes in a standard-compliant test environment, cannot drink water and drink, with the forearm exposed, remaining relaxed, avoiding touching the test area. The test was performed according to the instructions of the trans-epidermal moisture loss tester Tewameter. When measuring TEWL, the instrument probe must be placed in the center of the test area. At the time of measurement, the average value of 10 continuous points of the TEWL value curve (the standard deviation should be less than 0.2 g/(h.m) 2 ) As a measurement value); initial values for each zone were measured (prior to sample use) and TEWL values were measured for the test and control zones after a set time. The set time was 7 days and 14 days. Testing of the same subject must be accomplished by the same tester using the same instrument. The test should be stopped immediately during the use of the sample, such as when adverse effects occur on the skin of the subject, and the subject should be properly treated and the adverse effects recorded.
(4) Calculation result
Descriptive statistics: descriptive statistics are performed on the measured values of each test area, including number, mean, standard deviation, minimum, median, maximum.
Differential analysis: the rates of change of the TEWL averages on the sample application side and control side were calculated separately, and then the differences between the spot application area and control area at different measurement times were statistically analyzed using the rates of change. If the test data are normally distributed, adopting a t-test method to carry out statistical analysis; if the test data are in non-normal distribution, a rank sum test method is adopted for statistical analysis. The statistical methods all use a two-tailed test with a test level α=0.05.
(5) Result determination
Positive results: compared with the control side, the change rate of the TEWL value of the sample smearing side is significantly different, and P is smaller than 0.05, so that the tested sample has an effect of promoting the recovery of the transepidermal water loss of the barrier damaged skin.
Negative results: compared with the control side, the change rate of the TEWL value of the sample smearing side is not significantly different by more than 0.05, which indicates that the tested sample has no promoting effect on the recovery of the transepidermal water loss of the barrier damaged skin.
The test results are shown in Table 5.
TABLE 5 TEWL value for the percutaneous Water loss of test samples at various times
Statistical analysis of the experimental results in table 5 shows that the transdermal water loss TEWL values of examples 1 to 16 are significantly reduced compared with 0 day for 7 days and 14 days, and the change rate is significantly different from that of the control group, and P <0.001. Description the compositions of examples 1 to 16 have a promoting effect on the recovery of the percutaneous moisture loss of the barrier damaged skin. And are classified into four grades, examples 1 to 5, examples 6 to 9, examples 10 to 12 and examples 13 to 16, respectively, and recovery of percutaneous moisture loss is from low to high. Whereas the rate of change of the TEWL values of the percutaneous moisture loss of comparative examples 1 to 6 was much higher than that of the control group, indicating that the skin barrier was damaged. The results of this repair efficacy test also further demonstrate the synergy between the ingredients and amounts of the formulation.
5. Human body whitening and freckle removing efficacy test
The method is tested according to the specific requirements of a method for testing the freckle removing and whitening efficacy of the cosmetic according to chapter 5 of the eighth edition 2015 of the cosmetic safety technical Specification (a method for testing the freckle removing and whitening efficacy of a ultraviolet-induced human skin blackening model).
(1) Test sample: the compositions of examples 1 to 16 were each added to a common emulsion external matrix to prepare test samples, wherein the compositions account for 5% of the total mass of the emulsion.
Negative control: blackened area blank.
(2) Subject selection: and selecting qualified subjects according to the inclusion and exclusion criteria, and ensuring that the number of the effective cases finally completed in each test area is not less than 30.
Inclusion criteria: 18-60 years old, healthy men or women; the ITA degree value of the skin color of the test part is 20-41 degrees; no allergic diseases, no allergic history of cosmetics or other external preparations; no medicine affecting the light sensitivity is used in the recent history of no light-sensitive disease; skin at the tested part should be free of pigmentation, inflammation, scar, pigmented nevus, and hair; a person capable of receiving the skin of the test area for tanning using an artificial light source; the test procedure can be understood, voluntarily taking part in the test and signing a written informed consent.
Exclusion criteria: pregnant or lactating women, or those who have a pregnancy preparation plan recently; skin history patients with psoriasis, eczema, atopic dermatitis, severe acne, etc.; for the last 1 month, anti-inflammatory drugs such as corticosteroids, etc. are orally taken or externally used; for about 2 months, orally or topically applying any product or drug that affects skin color (e.g., hydroquinone-based formulations); the skin blackening marks of the test parts do not completely fade after the similar test is carried out in 3 months or before the similar test is carried out in 3 months; other clinical trials were enrolled in approximately 2 months; other clinical evaluations were considered unsuitable for the participants.
(3) Measurement procedure
Test instrument: adopting a xenon arc lamp sunlight simulator which has continuous spectrum radiation and can generate ultraviolet rays with UVA and UVB wavelengths, removing the wavelength below 290nm by using a proper filtering system, and performing metering verification or calibration on the output spectrum; a skin colorimeter having color space data defined by the international commission on illumination (CIE) that can be measured; a skin melanin detector having a principle of spectral absorption for detecting MI values of skin.
Environmental conditions: the instrument test links in the test process are all carried out under the environment that the temperature is 21+/-1 ℃ and the relative humidity is 50+/-10%RH.
Test procedure: and the qualified subjects enter a stage of establishing a human skin blackening model. The MED of the test site of each subject should be determined first, then each test zone should be selected at the test site, and irradiated with a solar simulator 1 time per day at the same irradiation point at a dose of 0.75 times the MED for 4 days continuously. The 4 days after the end of irradiation are skin darkening periods without any treatment. On day 5 after the irradiation, skin color instrument detection is carried out on the skin color of each test area, and the test area with poor consistency (the area with ITA degree value differing from the average value of all the test areas by more than 5) is removed. The corresponding test subjects were smeared according to a random table at each blackening test zone beginning the day. The test subjects were continuously applied for at least 4 weeks, and skin color was measured with the instrument 1 week, 2 weeks, 3 weeks, and 4 weeks after application and recorded. The results are shown in Table 6.
Data analysis: statistical analysis of the data was performed using statistical analysis software. The metering data are expressed as: the mean value is +/-standard deviation, normal distribution inspection is carried out, normal distribution requirements are met, paired t inspection is adopted for comparison before and after the mean value is self, and otherwise, two related sample rank sum inspection is adopted; comparing the grade data before and after use, and adopting two related sample ranks and tests; the comparison between the test area and the control area adopts independent sample t test or rank sum test; meanwhile, the regression coefficient (slope k value) of each parameter with time was calculated, and the significance level was P <0.05.
Conclusion of the test: the difference of the skin color visual scores or the ITA degree difference of the test sample at any time point before and after the application is obviously improved compared with the negative control (P < 0.05), or when the regression coefficient analysis integrally judges that the skin blackening of the test sample is obviously improved compared with the negative control (P < 0.05), the test sample is identified to have the efficacy of removing spots and whitening, otherwise, the test sample is identified to have no efficacy of removing spots and whitening.
TABLE 6 ITA values and MI values at various times after sample use
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Note that: the larger the ITA degree value is, the lighter the skin color is, otherwise, the darker the skin color is; the smaller the MI value, the lower the skin melanin content, and conversely the higher the skin melanin content.
Statistical analysis of the experimental results of table 6, the ITA ° values of examples 1-16 were significantly improved compared to 0 weeks using 1 week, 2 weeks, 3 weeks, 4 weeks, with significant differences, P <0.01, compared to the negative control; the MI values of examples 1-16 were significantly reduced compared to 0 weeks using 1 week, 2 weeks, 3 weeks, 4 weeks, with differences significantly different from the negative control, P <0.01. The compositions of examples 1 to 16 are shown to have freckle-removing and whitening effects, and are classified into four classes, examples 1 to 5, examples 6 to 9, examples 10 to 12 and examples 13 to 16, respectively, and the effects of skin whitening are compared from low to high, which are caused by the different components of the compositions.
6. Chick embryo chorioallantoic membrane (CAMVA) experiment
Test sample: the compositions of examples 1 to 16 and comparative examples 1 to 6 were each added to a general emulsion external matrix to prepare test samples, wherein the compositions account for 5% of the total mass of the emulsion.
The testing method comprises the following steps: the chick embryo is placed in a microcomputer full-automatic incubator for incubation, a small hole is drilled at the narrow end of the chick embryo on the 4 th day, and about 4mL egg white is extracted. A window of about 2cm is opened above the chicken ring position, and the chicken ring is covered by a preservative film. Then placing the mixture into a constant temperature and humidity box for continuous incubation. On day 10, chick embryos are taken out, eggshells around a window are stripped, 226 chick embryos with good growth are selected and randomly divided into 24 groups, namely 3 negative control groups (blank), 3 positive control groups (common emulsion matrix for external use), test samples (10 pieces each) prepared by the compositions of examples 1-16 and comparative examples 1-6 are exposed to chorionic blood vessels, teflon rings are placed at the positions where blood vessels are dense, one Teflon ring is placed on each chick embryo, 40 mu L of each group of samples are added into each ring, then the chick embryos are placed into a constant temperature and humidity box for incubation for 30 minutes, and the Teflon rings are taken off and washed by PBS until no obvious residues exist in the test samples. The vascular injury in the teflon ring was observed under a stereoscopic microscope using the area outside the teflon ring as a control, and the vascular stimulation effect was scored according to table 7. The results are shown in Table 8.
TABLE 7 CAMVA test scoring criteria
TABLE 8 CAMVA test scoring table
| Sample of | Score of |
| Negative control group | Are all 0 level |
| Positive control group | Are all 0 level |
| Example 1 | Are all 0 level |
| Example 2 | Are all 0 level |
| Example 3 | Are all 0 level |
| Example 4 | Are all 0 level |
| Example 5 | Are all 0 level |
| Example 6 | Are all 0 level |
| Example 7 | Are all 0 level |
| Example 8 | Are all 0 level |
| Example 9 | Are all 0 level |
| Example 10 | Are all 0 level |
| Example 11 | Are all 0 level |
| Example 12 | Are all 0 level |
| Example 13 | Are all 0 level |
| Example 14 | Are all 0 level |
| Example 15 | Are all 0 level |
| Example 16 | Are all 0 level |
| Comparative example 1 | 2 stages 0, 4 stages 1, 3 stages 2, 1 stage 3 |
| Comparative example 2 | 3 stages 0, 4 stages 1, 3 stages 2 |
| Comparative example 3 | 1 level 0, 3 level 1, 3 level 2, 3 level 3 |
| Comparative example 4 | 4 stages 1, 3 stages 2, 3 stages 3 |
| Comparative example 5 | Are all 0 level |
| Comparative example 6 | Are all 0 level |
As can be seen from the CAMVA test results of Table 8, the compositions of examples 1 to 16 achieved the expected effect, all of which were grade 0. The compositions of comparative examples 1-4, however, present significant irritation potential. This is caused by the difference in the components and proportions of the compositions of comparative examples 1 to 6.
In summary, according to the invention, through the synergistic effect among the active substances of the snow carnosine, the nine peptide-1, the carnosine, the vitamin C, the copper gluconate, the tripeptide-1 and the glutathione, the results of the combination stability test, the efficacy test and the safety test can be seen that the examples have better performance in the aspect of whitening efficacy and are classified into 4-grade whitening grades, and the melanin inhibition rate of the examples and the comparative examples is more than 40%, but the stability of the comparative examples is poor; in the human body safety test, the results of the examples are negative, the safety is qualified, and the comparative examples have great potential safety hazards; in the repair efficacy test, the examples have an effect of promoting recovery of the percutaneous moisture loss of the barrier damaged skin, whereas the comparative examples may cause damage to the skin; the results of the chick embryo chorioallantoic membrane experiments also show that the comparative examples have different degrees of irritation hidden dangers to the skin, and the examples are all 0 grade and have no irritation to the skin. Therefore, the invention has the effects of whitening, removing freckles and protecting skin barriers on the basis of ensuring that the safe skin of a human body is not damaged.
Claims (3)
1. The peptide composition with multiple whitening and freckle removing effects is characterized by comprising the following components in parts by weight: 5 to 15 parts of snow carnosine, 2 to 7 parts of nonapeptide-1, 1 to 10 parts of carnosine, 3 to 8 parts of vitamin C, 1 to 5 parts of copper gluconate, 1 to 7 parts of tripeptide-1 and 2 to 8 parts of glutathione.
2. The peptide composition with multiple whitening and freckle removing effects according to claim 1, which is characterized by comprising the following components in parts by weight: 8-11 parts of snow carnosine, 4-5 parts of nonapeptide-1, 5-7 parts of carnosine, 5-6 parts of vitamin C, 2-4 parts of copper gluconate, 3-6 parts of tripeptide-1 and 4-7 parts of glutathione.
3. The use of the peptide composition with multiple whitening and freckle removing effects of claim 1 in the preparation of skin care product additives.
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| WO2005048972A1 (en) * | 2003-11-19 | 2005-06-02 | Amorepacific Corporation | Skin whitening cosmetic composition |
| KR20100009444A (en) * | 2008-07-18 | 2010-01-27 | (주)바이오에프디엔씨 | Cosmetic composition for skin-whitening and antibacterial activity and method for preparing the same |
| CN107375005A (en) * | 2017-08-07 | 2017-11-24 | 深圳市维琪医药研发有限公司 | A kind of antioxidation and peptide composition and its application for suppressing B16 cell |
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