Method for protecting keratinous substances from carbonylation stimuli by using an anti-carbonylation composition and a composition containing polylysine

Abstract

Compositions are provided herein that reduce and / or inhibit protein carbonylation and contain polylysine. By using a composition containing polylysine, a method of protecting the skin from carbonylation stimuli is provided, as well as a non-therapeutic cosmetic method for caring for keratinous substances, particularly the skin, by applying a composition that reduces and / or inhibits protein carbonylation and contains polylysine.

Description

Technical Field

[0001] The present application relates to an anti-carbonylation composition containing polylysine, and to a method for reducing and / or inhibiting protein carbonylation by using a composition containing polylysine, and more particularly to a method for protecting tissues (such as keratinous substances, particularly keratin fibers, such as skin, etc.) from carbonylation stimuli.

Background Art

[0002] The skin is the body's first barrier against external environmental stimuli. An important mechanism for environmental pollutants that penetrate the skin, such as particulate matter, ultraviolet rays, ozone, and the like, is to attack unsaturated lipids in the surface layer of the skin by oxidative damage, destroy the outermost protective layer of the skin, and produce reactive carbonyl substances, such as malondialdehyde, 4-hydroxynonenal, acrolein, etc. When the content of these types of substances exceeds the body's removal ability, carbonylation stimuli occur, which induce carbonylation modification of biopolymers, such as proteins, resulting in structural changes and loss of function.

[0003] During protein carbonylation, reactive aldehydes or ketones are introduced into the protein by oxidation. Protein carbonyls have been shown to be a major product of protein oxidation and may be formed by oxidative cleavage of proteins, direct oxidation of amino acid residues, or covalent bond reactions with aldehydes derived from lipid peroxidation.

[0004] Protein carbonylation refers to the process in which amino or imino groups in the side chains of amino acid residues are attacked by oxygen free radicals and finally converted into aldehyde groups, releasing NH3+, where the amino acids in the protein side chains are attacked by hydroxyl free radicals and modified by oxidation, and then the peptide bond is broken, a carbonyl group is generated at the cleavage point, and it is generally understood that the content of the carbonyl group increases significantly.

[0005] In particular, protein carbonylation occurs by direct metal-catalyzed oxidation of amino acid side chains (primary protein carbonylation) or by addition of reactive aldehydes to amino acid side chains (secondary protein carbonylation). As such, reactive aldehydes of external origin, such as acrolein, are well known to be derived from tobacco smoke.

[0006] It has previously been reported that primary protein carbonylation plays a role in the mechanism of reactive oxygen species (ROS) signaling. Reactive oxygen species (ROS) in the skin may change proteins in the cellular structure of the skin. If reactive oxygen species (ROS) are not completely controlled by the skin antioxidant defense system, proteins in the skin will be carbonylated, resulting in skin damage.

[0007] It has been well documented that the production of carbonyl proteins is characteristic of carbonylation stress, for example, induced by ultraviolet radiation or the external application of oxidative chemicals, either as an external factor or as a chemical attack by reactive carbonyls derived from internal factors, such as the decomposition of lipid peroxides.

[0008] Carbonylated proteins in the skin are mainly present in the epidermis and dermis, and the content of carbonylated proteins in the epidermis is greater than that in the dermis. This is because the epidermis of the skin is continuously exposed to an oxidative environment and an unsaturated lipid environment secreted by sebaceous glands, which create favorable conditions for the continuous generation of reactive aldehyde compounds.

[0009] Furthermore, carbonyl stress causes skin damage, such as accelerating skin aging, or various pathological reactions of the skin. Protein carbonylation impairs the water retention capacity of the skin, affects the light transmittance of the skin, and changes the optical properties of the skin, etc. In particular, the presence of carbonylated proteins is associated with changes in skin properties, including mechanical properties, a decrease in water content and an increase in epidermal water loss, dull skin, and a decrease in skin transparency. At the same time, an increase in the level of carbonylated proteins may lead to inflammatory skin diseases, such as psoriasis and dermatitis.

Summary of the Invention

Problems to be Solved by the Invention

[0010] Therefore, there is a great need for anti-carbonylation agents that can inhibit or prevent protein carbonylation, as well as skin care compositions and methods containing anti-carbonylation agents for improving the appearance of the skin or at least one symptom of skin aging (such as a decrease in water retention capacity, dull skin, skin yellowness, a decrease in skin transparency, elasticity, light and shade, etc.) and reducing the occurrence of inflammatory skin diseases, such as psoriasis and dermatitis.

Means for Solving the Problems

[0011] The present application aims at a cosmetic composition containing at least one polylysine. The present application also aims at a method for reducing or inhibiting protein carbonylation against keratinous substances, particularly against the skin.

[0012] Throughout this specification, including the claims, the term "comprising a" should be understood to be synonymous with "comprising at least one" unless otherwise stated. Furthermore, the expression "at least one" used in this specification is equivalent to the expression "one or more".

[0013] By "local application", it is intended that the composition be applied or spread on the surface of a keratinous substance, for example at least one area of the skin.

[0014] As used herein, "anti-carbonylation" means reducing and / or inhibiting protein carbonylation, and in particular protecting tissues (such as keratinous substances, especially keratin fibers, such as skin, etc.) from carbonylation stimuli.

[0015] As used herein, "polylysine" means a lysine-based polymer.

[0016] It has unexpectedly been found that an anti-carbonylation composition (or agent) of the invention comprising (or consisting of) at least one polylysine can achieve improved anti-carbonylation effectiveness. The present application aims at an anti-carbonylation composition (or agent) of the invention comprising (or consisting of) at least one polylysine as a cosmetic composition. The present application also aims at a method for reducing and / or inhibiting protein carbonylation on keratinous substances, especially keratin fibers, such as skin, by applying a composition comprising an anti-carbonylation composition (or agent) of the invention comprising (or consisting of) at least one polylysine.

[0017] In one embodiment, the polylysine is preferably an L-lysine-based polymer.

[0018] In another embodiment, the polylysine may be selected from the group consisting of linear α-polylysine, linear ε-polylysine, highly branched polylysine or dendritic polylysine.

[0019] In one embodiment, the polylysine is preferably ε-polylysine and / or dendritic polylysine.

[0020] In other embodiments, the polylysine has a molecular weight of more than 200, more preferably more than 300, or more preferably more than 400, preferably more than 500, or more preferably more than 800. In particular, the polylysine has a molecular weight of less than 200,000, more preferably less than 150,000, or more preferably less than 100,000, preferably less than 50,000, or more preferably less than 20,000. For example, the polylysine has a molecular weight of 200 to 200,000, more preferably 400 to 100,000, or 600 to 50,000, or 800 to 20,000, or 1,000 to 15,000, preferably 1,000 to 10,000. When the polylysine is α-polylysine, the preferred molecular weight of α-polylysine is less than 30,000.

[0021] In other embodiments, the polylysine is present in the cosmetic composition in an amount of 0.0001% to 10% by weight, preferably 0.001% to 5% by weight, preferably 0.001% to 2% by weight, based on the total weight of the composition.

[0022] In other embodiments, the composition of the present invention further comprises a functional agent, such as at least one active agent, such as a humectant, such as a protein hydrolyzate, and a polyol, such as glycerol, glycol, polyethylene glycol, and sugar derivatives; natural extracts; vitamins, such as vitamin A (retinol), vitamin E (tocopherol), vitamin C (ascorbic acid), vitamin B5 (panthenol), vitamin B3 (niacinamide), derivatives (especially esters) of these vitamins and mixtures thereof; urea; caffeine; salicylic acid and its derivatives; alpha-hydroxy acids, such as lactic acid and glycolic acid, and their derivatives; retinoids, such as carotenoids and vitamin A derivatives; sunscreen agents; essential oils of mint, aloe vera or carrot and mixtures thereof, and may be a skin care agent selected from the group consisting of, and further comprises a functional agent.

[0023] Another subject of the present application is a method for preparing an anti-carbonylation composition, comprising the following steps: - (1) Optionally preparing a formula base and then cooling it, for example, cooling it to room temperature. - (2) For example, in water, a step of dispersing the obtained polylysine and - (3) 1) A step of mixing a formula base and 2) a polylysine (s) solution at room temperature is to provide a method including the above steps.

[0024] Another subject of this application is to provide a non-therapeutic cosmetic method for combating the denaturation of keratinous substances, particularly the skin, by applying the composition of the present invention.

[0025] Another subject of this application is to provide a non-therapeutic cosmetic method for caring for keratinous substances, particularly the skin, by applying the anti-carbonylation composition of the present invention.

[0026] Another subject of this application relates to the use of the composition of the present invention for preparing a product for caring for keratinous substances, particularly the skin.

[0027] Another subject of this application is to provide a cosmetic containing the composition of the present invention.

[0028] Other characteristics and advantages of this application should become clearer upon reading this specification and the following examples.

[0029] In this application, unless otherwise specifically mentioned, contents, parts and percentages are expressed on a weight basis.

Mode for Carrying Out the Invention

[0030] Polylysine Polylysine refers to several types of lysine-based homopolymers, which may differ from each other in terms of stereochemistry and bonding position. Lysine (an essential amino acid for humans) is the primary building block of polylysine and is available in two chiral forms: L-lysine and D-lysine. The polymerization of specific chiral lysine monomers results in L-polylysine and D-polylysine, respectively. The precursor amino acid lysine contains two amino groups, one on the α-carbon and one on the ε-carbon. Both can serve as the site of polymerization, resulting in α-polylysine or ε-polylysine. Polylysine can be further classified into linear, highly branched, and dendritic polylysine according to the topology of the polymer chain. The molecular weight of polylysine can range from 100 to 1,000,000.

[0031] In this application, polylysine refers to lysine-based polymers such as linear ε-polylysine (ε-PL), linear α-polylysine (α-PL), highly branched polylysine, and dendritic polylysine.

[0032] Typically, polylysine is present in topical compositions at the following concentrations: 0.0001 wt% to 10 wt%, preferably 0.001 wt% to 5 wt%, preferably 0.001 wt% to 2 wt% based on the total weight of the composition.

[0033]

Chemical formula

[0034] Surprisingly, without wishing to be bound by theory, the Applicant has found that polylysine, at its concentration in the range of, for example, 0.005% to 0.05% in some cases, exceeds monomeric lysine in anti-carbonylation effectiveness, and this may contribute to the "multivalent effect" of the polymer chain.

[0035] Surprisingly, without wishing to be bound by theory, polylysine is preferably selected from ε-polylysine and / or dendritic polylysine.

[0036] Active agent According to one embodiment of the present application, the product of the present application comprises at least one functional agent, for example, a peeling or moisturizing agent, a depigmenting or anti-depigmenting agent, an anti-glycation agent, an anti-NO agent, an agent for stimulating the synthesis of skin or epidermal polymers and / or for preventing their degradation, an agent for stimulating fibroblast or keratinocyte proliferation and / or keratinocyte differentiation, a muscle relaxant or a dermo-decontracting agent, a free radical scavenger or an anti-staining agent, a tensioning agent, an agent acting on capillary circulation, and in particular the following particularly active agents, and may include at least one active agent selected from the group consisting of mixtures thereof.

[0037] Among all the active agents that can be used in the present application, particular mention may be made of α- or β-hydroxy acids, such as lactic acid, glycolic acid, citric acid, 5-octanoylsalicylic acid, α-hydroxydecanoic acid, α-hydroxylauric acid, tartaric acid, glucuronic acid, galacturonic acid, acrylic acid, α-hydroxybutyric acid, α-hydroxyisobutyric acid, malic acid, mandelic acid, phosphoric acid, pyruvic acid, lactobionic acid, and salicylic acid.

[0038] Anti-acne agents, such as salicylic acid or benzoyl peroxide, octopirox, d- and l-sulfur-containing amino acids, their salts, and their N-acetyl derivatives, such as N-acetylcysteine, or agents aimed at preventing and / or improving skin aging, such as the aforementioned α- and β-hydroxy acids, retinoids such as retinoic acid, retinol, and its esters, such as retinol propionate, and retinol acetate, or retinol palmitate, niacinamide, allantoin, aloe extract, azelaic acid, bisabolol, phytic acid, collagen, or agents that stimulate collagen formation, vitamins such as vitamin C or its derivatives, such as ascorbyl glucoside, vitamin E or its derivatives, vitamin A or its derivatives, vitamin F or its derivatives, the aforementioned d- and l-sulfur-containing amino acids and their derivatives, elastin, N-acetyl-D-glucosamine, luteolin, or antioxidants, such as green tea or its active fraction, glycerin, laponite, caffeine, aromatic essential oils, colorants, free radical scavengers, moisturizers, depigmenting agents, agents for improving skin color, such as dihydroxyacetone or tyrosine ester type self-tanning agents, lipid regulators, emollients, anti-wrinkle agents, keratolytic agents, fresheners, deodorants, anesthetics, nutrients, and mixtures thereof can also be used. Bleaching agents, such as kojic acid, ascorbyl phosphate, ascorbyl glucoside, ascorbic acid, and mixtures thereof can also be used.

[0039] In the case of masks, active agents for improving skin conditions, such as moisturizers or agents useful for improving the natural lipid barrier, such as ceramides, cholesterol sulfate, and / or fatty acids, and mixtures thereof can also be used. Enzymes having activity on the skin, such as proteases, lipases, cerebrosidases, and / or melanase, and mixtures thereof can also be used.

[0040] As other examples of active agents that may be suitable for practicing the present application, there are agents for drugs, peptides, proteins, detectable labels, contrast reagents, analgesics, anesthetics, antibacterial agents, anti-yeast agents, antifungal agents, antiviral agents, anti-inflammatory agents for the skin, antipruritics, antiemetics, vasoprotective agents, agents for motion sickness, anti-irritants, anti-inflammatory agents, immunomodulators, anti-hyperkeratolytic agents, agents for treating dry skin, antiperspirants, anti-psoriatic agents, anti-dandruff agents, anti-aging agents, anti-asthmatic and bronchodilator agents, sunscreens, antihistamines, healing agents, corticosteroids, tanning agents, and mixtures thereof.

[0041] The content of at least one active agent in the composition may be adjusted according to the intended purpose of the composition.

[0042] Auxiliary agents In a known manner, the compositions of the present application may also contain auxiliary agents common in cosmetics and / or dermatology, such as preservatives, antioxidants, pH adjusters (acidic or basic), fragrances, fillers, bactericides, odor absorbers, colorants (pigments and dyes), emulsifiers, and lipid vesicles.

[0043] Needless to say, those skilled in the art will take care to select this or these optional additional compound(s) and / or their amounts such that the benefits of the compositions according to the present application are not or are substantially not adversely affected by the intended addition.

[0044] The numerical ranges and parameters indicating the broad scope of the present application are approximations, but the numerical values reported in specific examples are reported as accurately as possible. However, any numerical values inherently contain certain errors resulting necessarily from the standard deviation found in their respective measurements. The following examples are intended to illustrate the present application without limiting the scope as a result.

Examples

[0045] The amounts / concentrations of the components in the compositions / formulas described below are expressed as parts by weight.

[0046] I. Evaluation of the Composition of the Present Invention 1. In vitro anti-carbonylation efficacy test This assay provides an in vitro method for determining how much an anti-carbonylation agent / composition affects the degree of protein carbonylation. Using deionized water, the anti-carbonylation agent / composition to be tested was made into a 1% solution, bovine serum albumin (BSA) into a 10 mg / mL solution, and acrolein into a 10 mM solution. Then, in a 1.5 mL centrifuge tube, according to the contents in Table 1, BSA (10 mg / mL), anti-carbonylation agent / composition (1%), acrolein (10 mM), and phosphate buffer solution (PBS) were added to the centrifuge tube. Samples containing only BSA and PBS functioned as the baseline, and samples containing BSA, acrolein, and PBS functioned as the blank. The final liquid volume in each centrifuge tube was 1.0 mL. Each group contained three replicate samples, and all samples were left standing at 37 °C overnight.

[0047] [Table 1]

[0048] The protein carbonyl content in the above sample was detected by using the Protein Carbonyl Assay Kit from Nanjing Jiancheng Bioengineering Institute. The detection method is as follows: Take 0.1 mL of the sample into an assay tube, then add 0.4 mL of Reagent 3 in the kit, vortex for 1 minute to mix, and react at 37 °C in the dark for 30 minutes. Add 0.5 mL of Reagent 5 in the kit, vortex for 1 minute to mix, centrifuge at 12,000 r / min for 10 minutes at 4 °C, discard the supernatant, and leave the pellet. Add 1.0 mL of a mixed solution of absolute ethanol and ethyl acetate (1:1), vortex for 1 minute to mix, centrifuge at 12,000 r / min for 10 minutes at 4 °C, discard the supernatant, leave the precipitate, and repeat the above steps 4 times. Add 1.25 mL of Reagent VI in the kit, mix well, place this at 37 °C for 15 minutes, vortex to mix, dissolve all the precipitate, and centrifuge at 12,000 r / min for 15 minutes. Take the supernatant into a quartz cuvette with a light path of 0.5 cm, zero with Reagent 6, and measure the absorbance value of each tube at 370 nm (ultraviolet). According to the formula, the applicants can obtain the protein carbonyl content.

[0049]

Number

[0050] The protein content of the sample was determined using the Thermos Micro BCA (trademark) protein assay kit. First, a diluted albumin (BSA) standard was prepared (working range = 20 - 750 μg / mL). BCA reagent A was mixed with reagent B (reagent A: reagent B = 50:1) to prepare the working solution. 25 μL of each of the protein standards at various dilution concentrations and the protein sample were taken and added to the microplate. 200 μL of the working solution was added to each well and mixed thoroughly by shaking on a shaker for 30 seconds. The microplate was sealed and incubated at 37 °C for 30 minutes. The microplate was cooled to room temperature and the absorbance of the sample at a wavelength of 562 nm was measured. The corrected absorbance value was obtained by subtracting the absorbance value of the blank sample from the reading values of each standard and sample. The protein concentration was read against the standard curve prepared with a known amount of BSA. Referring to the standard curve, the protein concentration of each sample within the linear range of the standard curve was read according to the corrected absorbance value of each protein sample. Based on the sample volume and dilution, the amount of protein in the original sample was calculated.

[0051] 2. Anti - carbonylation effectiveness of polylysine The anti - carbonylation ability of each anti - carbonylation agent / composition can be obtained by dividing the degree of carbonylation of the sample by the BSA protein content. The results are shown in Tables 2 and 3 below.

[0052]

Table 2

[0053]

Table 3

[0054] Polylysine reduces and / or inhibits protein carbonylation as shown above.

[0055] II. Formulation examples The composition of the present invention containing polylysine is prepared by sufficiently mixing these components as follows.

[0056] [Table 4]

[0057] III. Sensory evaluation of the anti-carbonylation composition The sensory properties of Compositions 1 to 4 were evaluated after applying them to the skin and are summarized in the following table:

[0058] [Table 5]

[0059] The cosmetic properties of Compositions 1 to 4 were good, especially with regard to spreadability, softness, and non-stickiness.

[0060] Although the present invention has been described in connection with preferred embodiments, it is not intended to limit the scope of the present invention to the specific forms shown. On the contrary, it is intended to cover alternatives, modifications, and equivalents that may fall within the spirit and scope of the present invention as defined by the appended claims.

Claims

Claim 1 An anti-carbonylation composition containing at least one kind of polylysine. Claim 2 The anti-carbonylation composition according to claim 1, wherein the polylysine is selected from the group consisting of at least one kind of lysine-based polymer. Claim 3 The anti-carbonylation composition according to any one of claims 1 and 2, wherein at least one kind of polylysine is an L-lysine-based polymer. Claim 4 The anti-carbonylation composition according to any one of claims 1 to 3, wherein at least one kind of polylysine is selected from the group consisting of α-polylysine, ε-polylysine, highly branched polylysine and dendritic polylysine, preferably selected from the group consisting of ε-polylysine and / or dendritic polylysine. Claim 5 The anti-carbonylation composition according to any one of claims 1 to 4, wherein at least one kind of polylysine has a molecular weight of more than 200, further more than 300, or more than 400, preferably more than 500, or more than 800, and in particular, the polylysine has a molecular weight of less than 200,000, further less than 150,000, or less than 100,000, preferably less than 50,000, or less than 20,000. For example, the polylysine has a molecular weight of 200 to 200,000, further 400 to 100,000, or 600 to 50,000, or 800 to 20,000, or 1,000 to 15,000, preferably 1,000 to 10,000. Claim 6 The anti-carbonylation composition according to any one of claims 1 to 5, wherein when the polylysine is α-polylysine, the molecular weight of α-polylysine is less than 30,000. Claim 7 The anti-carbonylation composition according to any one of claims 1 to 6, wherein at least one kind of polylysine is present in the composition in an amount of 0.0001% by weight to 10% by weight, preferably 0.001% by weight to 5% by weight, preferably 0.001% by weight to 2% by weight based on the total weight of the composition. Claim 8 At least one functional agent, such as at least one active agent, preferably a humectant, such as a protein hydrolyzate, and a polyol, such as glycerol, glycol, such as polyethylene glycol, and a sugar derivative; a natural extract; vitamins, such as vitamin A (retinol), vitamin E (tocopherol), vitamin C (ascorbic acid), vitamin B5 (panthenol), vitamin B3 (niacinamide), derivatives (especially esters) of these vitamins and mixtures thereof; urea; caffeine; salicylic acid and its derivatives; alpha-hydroxy acids, such as lactic acid and glycolic acid, and their derivatives; retinoids, such as carotenoids and vitamin A derivatives; sunscreen agents; mint, aloe vera or lavender essential oils and mixtures thereof, the anti-carbonylation composition according to any one of claims 1 to 7, further comprising a skin care agent selected from the group consisting of.

9. A cosmetic comprising the anti-carbonylation composition according to any one of claims 1 to 8.

10. A method for preparing the anti-carbonylation composition according to any one of claims 1 to 8 or the cosmetic according to claim 9, comprising the following steps: - (1) Optionally preparing a formula base and then cooling it, for example cooling it to room temperature. - (2) Dispersing polylysine (plural available) in, for example, water. And - (3) Mixing 1) the formula base and 2) the polylysine (plural available) solution at room temperature. A method comprising.

11. A non-therapeutic cosmetic method for reducing and / or inhibiting protein carbonylation of keratinous substances, particularly the skin, by applying the anti-carbonylation composition according to any one of claims 1 to 8 or the cosmetic according to claim 9.

12. A non-therapeutic cosmetic method for caring for keratinous substances, particularly the skin, by applying the anti-carbonylation composition according to any one of claims 1 to 8 or the cosmetic according to claim 9.

13. Use of the composition according to any one of claims 1 to 8 or the cosmetic according to claim 9 for preparing a product for caring for keratinous substances, particularly the skin.

14. Use of the composition according to any one of claims 1 to 8 or the cosmetic according to claim 9 for preparing a product that reduces or prevents protein carbonylation, particularly with respect to keratinous substances, on the skin.

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