Anti-carbonylation compositions and uses thereof

Abstract

The present invention relates to anti-carbonylation compositions comprising at least one anti-carbonylation active, to methods of reducing and / or inhibiting carbonylation of proteins, in particular to methods of protecting tissues (e.g., keratin materials, in particular keratin fibres, such as skin, etc.) from carbonylation stimulation by using said anti-carbonylation compositions, and to methods of reducing and / or inhibiting carbonylation of proteins. And the use of the selected hyperbranched polymer as an anti-carbonylation active.

Description

Technical Field

[0001] The present invention relates to an anti-carbonylation composition comprising at least one anti-carbonylation active substance, a method for reducing and / or inhibiting protein carbonylation, in particular a method for protecting tissues (e.g., keratin materials, especially keratin fibers, such as skin, etc.) from carbonylation stimulation by using the anti-carbonylation composition, and the use of selected hyperbranched polymers as anti-carbonylation active substances. Background Art

[0002] The skin is the body's first barrier against external environmental stimuli. An important mechanism by which environmental pollutants (such as particulate matter, ultraviolet rays, ozone, etc.) invade the skin is to attack the unsaturated lipids in the surface layer of the skin through oxidative damage, destroy the outermost protective layer of the skin, and produce active carbonyl substances, such as malondialdehyde, 4-hydroxy nonenal, acrolein, etc. When the content of these types of substances exceeds the body's clearance capacity, carbonyl stimulation will occur, which induces carbonyl modification of biological macromolecules (such as proteins), resulting in structural changes and loss of function.

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

[0004] Protein carbonylation is generally considered to be the process in which the amino or imino groups in the side chains of amino acid residues are attacked by oxygen free radicals and eventually converted into aldehyde groups, releasing NH4 + or R-NH3 + The process in which the peptide bond breaks and carbonyl groups are generated at the break. After the protein side chain amino acids are attacked by hydroxyl free radicals and oxidatively modified, the carbonyl content increases greatly.

[0005] Specifically, protein carbonylation occurs due to direct metal-catalyzed oxidation of amino acid side chains (primary protein carbonylation) or addition of reactive aldehydes to amino acid side chains (secondary protein carbonylation). As such, it is well known that exogenous reactive aldehydes (such as acrolein) are derived from cigarette smoke.

[0006] It has been previously reported that primary protein carbonylation plays a role in the reactive oxygen species (ROS) signaling mechanism. ROS in the skin can alter proteins in the skin cell structure. If ROS are not fully controlled by the skin's antioxidant defense system, proteins in the skin will become carbonylated, leading to skin damage.

[0007] It is well documented that the production of carbonyl proteins is a hallmark of carbonylation stress, e.g. due to exogenous factors, such as induced by UV radiation or topical oxidative chemicals, or due to endogenous factors, such as chemical attack by reactive carbonyls derived from lipid peroxidation degradation.

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

[0009] In addition, carbonyl stress leads to skin damage (e.g., accelerated skin aging), or various pathological reactions of the skin. Protein carbonylation impairs the skin's ability to retain water, affects the skin's light transmittance, changes the skin's optical properties, etc. Specifically, the presence of carbonylated proteins is associated with changes in skin properties: such as mechanical properties; water retention capacity, including decreased water content and increased epidermal water loss; dull skin; and decreased skin transparency. At the same time, increased levels of carbonylated proteins may lead to inflammatory skin diseases such as psoriasis and dermatitis.

[0010] Therefore, there is a great need for anti-carbonylating agents that can inhibit or prevent protein carbonylation, as well as skin care compositions and methods comprising anti-carbonylating agents, to improve the appearance of the skin or improve at least one sign of skin aging (e.g., reduced water retention capacity, dull skin, yellowing of the skin, reduced skin clarity, elasticity, contrast, etc.), and to reduce the occurrence of inflammatory skin diseases (e.g., psoriasis and dermatitis). Summary of the invention

[0011] The present inventors have now discovered that the above-mentioned object can be achieved by the following embodiments.

[0012] One subject of the present invention is to provide an anti-carbonylation composition comprising at least one anti-carbonylation active substance, wherein the anti-carbonylation active substance comprises in its structural formula at least four units, and wherein the surface primary amino groups constitute greater than about 8 weight percent of the molecular weight of the anti-carbonylation active.

[0013] In one embodiment, the anti-carbonylation active is multi-branched.

[0014] In a preferred embodiment, the anti-carbonylation active agent has a molecular weight of about 500 or more, preferably about 800 or more, more preferably about 1000 or more, and most preferably about 1200 or more. In other preferred embodiments, the anti-carbonylation active has a molecular weight of about 120,000 or less, preferably about 60,000 or less, more preferably about 10,000 or less, and most preferably about 5,000 or less. In yet other embodiments, the anti-carbonylation active has a molecular weight of 500 to 120,000, preferably 800 to 60,000, more preferably 1,000 to 10,000, and most preferably 1,200 to 5,000.

[0015] In a specific embodiment, the anti-carbonylation active has a molecular weight of about 500 or more, preferably about 800 or more, more preferably about 1000 or more, most preferably about 1200 or more, and about 120,000 or less, preferably about 60,000 or less, more preferably about 10,000 or less, most preferably about 5,000 or less; preferably, the anti-carbonylation active has a molecular weight of 500 to 120,000, preferably 800 to 60,000, more preferably 1000 to 10,000, most preferably 1200 to 5,000.

[0016] In one embodiment, the anti-carbonylation active has the structure of Formula I,

[0017]

[0018] Where R 1 -R 4 Each independently

[0019] R 5 and R 6 Each independently selected from H and

[0020] R 7 and R 8 Each independently selected from H and

[0021] R 9 and R 10 Each independently selected from H and

[0022] R 11 and R 12 Each independently selected from H and

[0023] R 13 and R14 Each independently selected from H and

[0024] n is selected from 0, 1, 2, 3 and 4;

[0025] R a and R b Each is independently selected from H, a substituted or unsubstituted C1-C3 alkyl group, wherein the substituted C1-C3 alkyl group contains one or more C1-C3 alkyl substituents, one or more ether bonds, one or more amine groups, or a combination thereof.

[0026] In a preferred embodiment, R 1 -R 4 Each independently

[0027] R 5 and R 6 Each independently selected from H and

[0028] R 7 and R 8 Each independently selected from H and

[0029] R 9 and R 10 Each independently selected from H and

[0030] n is selected from 0, 1 and 2;

[0031] R a and R b It's H.

[0032] In a further preferred embodiment, the anti-carbonylation active substance may be selected from

[0033] and combinations thereof.

[0034] In one embodiment, the anti-carbonylation active is present in the composition in an amount of about 0.0001 wt % to about 10 wt %, preferably about 0.001 wt % to about 5 wt %, preferably about 0.005 wt % to about 2 wt %, relative to the total weight of the anti-carbonylation composition.

[0035] In another embodiment, the composition of the present invention further comprises a functional agent, which may be, for example, at least one active agent, such as a skin care agent, selected from moisturizers (e.g., protein hydrolyzates), and polyols (e.g., glycerol), glycols, polyethylene glycols, 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 of these vitamins (especially esters), 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; sunscreens; peppermint essential oil, aloe essential oil, or ginseng essential oil, and mixtures thereof.

[0036] Another subject of the present invention is to provide a method for preparing the anti-carbonylation composition, which comprises the following steps:

[0037] - (1) optionally, providing a formula base, which is then cooled, for example to room temperature,

[0038] - (2) dispersing at least one anti-carbonylation active in a cosmetically acceptable vehicle such as water,

[0039] as well as

[0040] - (3) 1) the formulation base and 2) the anti-carbonylation active solution are mixed at room temperature.

[0041] Another subject of the present invention is a cosmetic product comprising the anti-carbonylation composition of the present invention.

[0042] Another subject of the present invention is to provide a non-therapeutic method for combating the degeneration of keratin materials, preferably the skin, more preferably the facial skin, by topically applying to said keratin materials an effective amount of an anti-carbonylation composition according to the invention or a cosmetic according to the invention.

[0043] Another subject of the present invention is to provide a non-therapeutic method for reducing and / or inhibiting protein carbonylation on keratin materials, preferably the skin, more preferably the facial skin, by topically applying an effective amount of the anti-carbonylation composition of the present invention or the cosmetic product of the present invention to said keratin materials.

[0044] Another subject of the present invention is to provide a non-therapeutic method for protecting keratin materials from carbonylation irritation by topically applying an effective amount of the anti-carbonylation composition of the present invention or the cosmetic product of the present invention to keratin materials, preferably the skin, more preferably the facial skin.

[0045] Another subject of the present invention relates to the non-therapeutic use of an anti-carbonylation composition according to the invention or a cosmetic product according to the invention for combating the degeneration of keratin materials, preferably the skin, more preferably the skin of the face.

[0046] Another subject of the present invention relates to the non-therapeutic use of an anti-carbonylation composition according to the invention or a cosmetic product according to the invention for reducing and / or inhibiting protein carbonylation on keratin materials, preferably on the skin, more preferably on the facial skin.

[0047] Another subject of the present invention relates to the non-therapeutic use of an anticarbonylation composition according to the invention or a cosmetic product according to the invention for protecting keratin materials, preferably the skin, more preferably the facial skin, against carbonylation irritation.

[0048] Another subject of the present invention is to provide an anti-carbonylation composition according to the invention or a cosmetic product according to the invention for combating degeneration of keratin materials, preferably the skin, more preferably the skin of the face, caused by carbonylation irritation.

[0049] Another subject of the present invention is to provide an anti-carbonylation composition according to the invention or a cosmetic product according to the invention for reducing and / or inhibiting protein carbonylation on keratin materials, preferably the skin, more preferably the facial skin.

[0050] Another subject of the present invention is to provide an anti-carbonylation composition according to the invention or a cosmetic according to the invention for protecting keratin materials, preferably the skin, more preferably the facial skin, from carbonylation irritation. Other advantages of the present invention will emerge more clearly on reading the following description and examples. DETAILED DESCRIPTION

[0051] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the invention belongs. When the definition of a term in this specification conflicts with the meaning commonly understood by those skilled in the art to which the invention belongs, the definition described herein shall apply.

[0052] Unless otherwise indicated, all numbers used in the specification and claims to represent the amount of ingredients, properties such as molecular weight, reaction conditions, etc. should be understood to be modified by the term "about" in all cases. When the term "about" is used in conjunction with a numerical value in this article, it is intended to include not only the numerical value, but also the value within a tolerance or range of ± 5% of the exact numerical value associated with the word "about". Although the numerical ranges and parameters setting forth the wide range of the present invention are approximate, the numerical values ​​set forth in the specific examples are intended to be accurately reported in view of the measurement methods. However, any numerical value inherently contains certain errors that are necessarily caused by the standard deviation present in its respective test measurements.

[0053] In addition, the ranges described in the present invention and claims are intended to specifically include the entire range, not just the endpoints. For example, the range of 0-10 described is intended to disclose all integers between 0-10, such as 1, 2, 3, 4, and any subranges formed by any two values ​​thereof, all fractions between 0-10, such as 1.5, 2.3, 4.57, 6.1113, and any subranges formed by any two values ​​thereof, as well as the endpoints 0 and 10.

[0054] Throughout the specification, including the claims, the term "comprising a" should be understood as being synonymous with "comprising atleast one", unless otherwise indicated. As used herein, the term "comprising" should be interpreted as including all specifically mentioned features as well as optional, additional, unspecified features. As used herein, the use of the term "comprising" also discloses embodiments in which features other than the specifically mentioned features are absent (i.e., "consisting of..."). Furthermore, the expression "at least one" as used in this specification is equivalent to the expression "one or more".

[0055] "Topical application" means applying or spreading the composition onto the surface of a keratin material, such as at least one area of ​​the skin.

[0056] As used herein, "anti-carbonylation" refers to reducing and / or inhibiting protein carbonylation, especially protecting tissues (eg, keratin materials, especially keratin fibers, such as skin, etc.) from carbonylation stimulation.

[0057] As used herein, the term "and / or," when used in a list of two or more items, means that any one of the listed items may be used alone, or any combination of two or more of the listed items may be used. For example, if a composition is described as containing components A, B, and / or C, the composition may contain A alone; B alone; C alone; a combination of A and B; a combination of A and C; a combination of B and C; or a combination of A, B, and C.

[0058] Unless otherwise indicated, all parts or percentages in the present invention refer to parts or percentages by weight and are based on the total weight of the composition.

[0059] Polyamidoamine (PAMAM)

[0060] Polyamidoamine (PAMAM) is a class of dendrimers that are highly branched three-dimensional macromolecules with a well-defined structure and multifunctional terminal surfaces. The nanoscale polymers consist of an ethylenediamine core, a repetitively branched amidoamine interior structure, and a primary amine terminal surface.

[0061]

[0062] PAMAM Structure

[0063]

[0064] It has been unexpectedly found that the anti-carbonylation composition (or anti-carbonylation agent) of the present invention comprising at least one PAMAM polymer as an anti-carbonylation active can achieve improved anti-carbonylation efficacy. PAMAM dendrimers having many amino groups can be used as sacrificial agents to protect the skin from carbonylation damage. In addition, PAMAM has been found to have an anti-glycation effect. Therefore, it can be used as an anti-glycation active to reduce or slow the generation of advanced glycation end products (AGEs), which are a group of complex compounds generated by non-enzymatic interactions between proteins and reducing sugars or lipids.

[0065] Without wishing to be bound by any particular theory, it is proposed herein that, when applied topically, PAMAM dendrimers can scavenge and neutralize reactive carbonyl species, preventing them from reacting with biomolecules in the skin. The amino groups in PAMAM dendrimers are able to form stable adducts with carbonyl compounds, effectively trapping them and removing them from the skin environment. Compared to small molecules with amino groups such as lysine, they exhibit much stronger anti-carbonylation efficacy due to the "multivalent effect".

[0066] Based on the above findings, the present invention relates to an anti-carbonylation composition comprising at least one polyamide-amine (PAMAM) polymer as an anti-carbonylation active. Preferably, the anti-carbonylation active is selected from G0 PAMAM, G1 PAMAM, G2 PAMAM, G3 PAMAM, G4 PAMAM, G5 PAMAM and mixtures thereof. More preferably, the at least one anti-carbonylation active is selected from G0 PAMAM, G1 PAMAM, G2 PAMAM, G3 PAMAM and mixtures thereof. The at least one anti-carbonylation active is most preferably G0 PAMAM, G1 PAMAM or mixtures thereof.

[0067] Typically, the PAMAM polymer is present in the anti-carbonylation composition at a concentration of about 0.0001 wt % to about 10 wt %, preferably about 0.001 wt % to about 5 wt %, more preferably about 0.005 wt % to about 2 wt %, relative to the total weight of the composition; for example, about 0.005 wt %, about 0.01 wt %, about 0.02 wt %, about 0.05 wt %, about 0.1 wt %, about 0.2 wt %, about 0.5 wt %, about 1 wt %, relative to the total weight of the composition.

[0068] In another embodiment, the present invention relates to a method for preparing the carbonylation resistant composition of the present invention, comprising the steps of:

[0069] - (1) optionally, providing a formulation base and then cooling it, for example to room temperature,

[0070] - (2) dispersing at least one PAMAM polymer in a cosmetically acceptable vehicle such as water,

[0071] - (3) Mix 1) formulation base and 2) PAMAM polymer solution at room temperature.

[0072] The vehicle that can be used in the present invention is pharmaceutically acceptable or cosmetically acceptable. Such vehicles include, but are not limited to, one or more lower aliphatic alcohols, aliphatic ethers, aliphatic esters, polyols, glycols, water, or any combination thereof.

[0073] In a specific embodiment, the vehicle used in the present invention includes purified water, polyacrylate crosspolymer-6, caprylyl glycol, phenoxyethanol, hexylene glycol, caprylyl glycol / phenoxyethanol / hexylene glycol or any combination thereof.

[0074] In another embodiment, the present invention relates to the use of a PAMAM polymer as an anti-carbonylation active or as an active ingredient in an anti-carbonylation reaction.

[0075] In another embodiment, the present invention relates to the use of PAMAM polymers as anti-carbonylation actives or as active ingredients to combat degeneration of keratin materials, preferably the skin, more preferably the facial skin.

[0076] In another embodiment, the present invention relates to the use of a PAMAM polymer as an anti-carbonylation active or as an effective ingredient for reducing and / or inhibiting protein carbonylation on keratinous materials, preferably the skin, more preferably the facial skin.

[0077] In another embodiment, the present invention relates to the use of a PAMAM polymer as an anti-carbonylation active or as an effective ingredient for protecting keratinous materials, preferably the skin, more preferably the facial skin, from carbonylation irritation.

[0078] In another embodiment, the present invention relates to the use of PAMAM polymers as anti-carbonylation actives or as active ingredients for the preparation of cosmetics that combat degeneration of keratin materials, preferably the skin, more preferably the skin of the face. In another embodiment, the present invention relates to the use of PAMAM polymers as anti-carbonylation actives for the preparation of cosmetics that reduce and / or inhibit protein carbonylation on keratin materials, preferably the skin, more preferably the skin of the face.

[0079] In another embodiment, the present invention relates to the use of PAMAM polymers as anti-carbonylation actives for the preparation of cosmetics for protecting keratin materials, preferably the skin, more preferably the facial skin, from carbonylation irritation.

[0080] In another embodiment, the present invention is directed to PAMAM polymers for use as anti-carbonylation actives or as active ingredients in anti-carbonylation reactions.

[0081] In another embodiment, the present invention relates to PAMAM polymers for use as anti-carbonylation actives or as active ingredients against degeneration of keratinous materials, preferably the skin, more preferably the facial skin.

[0082] In another embodiment, the present invention is directed to PAMAM polymers for use as anti-carbonylation actives that reduce and / or inhibit protein carbonylation on keratinous materials, preferably the skin, more preferably the facial skin.

[0083] In another embodiment, the present invention is directed to PAMAM polymers for use as anti-carbonylation actives for protecting keratin materials, preferably skin, more preferably facial skin, from carbonylation irritation.

[0084] In another embodiment, the present invention relates to the use of a PAMAM polymer as an anti-glycation active or as an effective ingredient in an anti-glycation reaction.

[0085] In another embodiment, the present invention relates to the use of a PAMAM polymer as an anti-glycation active substance or as an effective ingredient for reducing or slowing down the generation of AGEs.

[0086] In another embodiment, the present invention relates to the use of PAMAM polymers as anti-glycation actives or as effective ingredients for protecting keratin materials, preferably the skin, more preferably the facial skin, from AGEs.

[0087] In another embodiment, the present invention relates to a PAMAM polymer for use as an anti-glycation active or as an effective ingredient in an anti-glycation reaction.

[0088] In another embodiment, the present invention relates to a PAMAM polymer for use as an anti-glycation active or as an effective ingredient for reducing or slowing down the generation of AGEs.

[0089] In another embodiment, the present invention relates to PAMAM polymers for use as anti-glycation actives or as effective ingredients for protecting keratin materials, preferably the skin, more preferably the facial skin, from AGEs.

[0090] Active Agent

[0091] According to one embodiment, the anti-carbonylation composition of the present invention may contain at least one functional agent, such as at least one active agent, for example selected from: desquamating agent or moisturizing agent; depigmenting agent or anti-depigmenting agent; anti-glycation agent; anti-NO agent; agents for stimulating the synthesis of dermal or epidermal macromolecules and / or for preventing their degradation; agents for stimulating fibroblast or keratinocyte proliferation and / or keratinocyte differentiation; muscle relaxants or skin relaxants (dermo-decontracting agent); free radical scavengers or anti-pollution agents; tensioning agents; agents acting on capillary circulation (capillary circulation); of course, in particular, the following active agents, and mixtures thereof.

[0092] Among all the active agents that can be used according to the invention, mention may be made in particular of: alpha- or beta-hydroxy acids, such as lactic acid, glycolic acid, citric acid, 5-octanoylsalicylic acid, alpha-hydroxydecanoic acid, alpha-hydroxylauric acid, tartaric acid, glucuronic acid, galacturonic acid, acrylic acid, alpha-hydroxybutyric acid, alpha-hydroxyisobutyric acid, malic acid, mandelic acid, phosphoric acid, pyruvic acid, lactobionic acid and salicylic acid.

[0093] Anti-acne agents may also be used, such as salicylic acid or benzoyl peroxide, octopirox, dextrorotatory and levorotatory sulfur-containing amino acids, their salts, and their N-acetyl derivatives (e.g. N-acetylcysteine), or agents that seek to prevent skin aging and / or improve its condition (e.g. the above-mentioned α- and β-hydroxy acids), retinoids (e.g. retinoic acid, retinol, and their esters, such as retinyl propionate and acetate, or retinyl palmitate), niacinamide, allantoin, aloe vera extract, azelaic acid, bisabolol, phytic acid, collagen, or agents that stimulate collagen formation, vitamins (e.g. vitamin C or its derivatives, such as ascorbyl glucoside, glucoside), vitamin E or its derivatives, vitamin A or its derivatives, vitamin F or its derivatives), right-handed and left-handed sulfur-containing amino acids and their derivatives as described above, elastin, N-acetyl D-glucosamine, luteolin, or antioxidants (e.g., green tea or its active fraction), glycerin, laponite, caffeine, essential aromatic oils, colorants, free radical scavengers, moisturizers, depigmenting agents, agents for improving skin color (e.g., artificial tanning agents of the dihydroxyacetone or tyrosine ester type), sebum regulators, softeners, anti-wrinkle agents, keratolytic agents, fresheners, deodorants, anesthetics, nutrients, and mixtures thereof. Bleaching agents such as kojic acid, ascorbyl phosphate, ascorbyl glucoside, ascorbic acid, and mixtures thereof may also be used.

[0094] In the case of a mask, active agents for improving skin conditions may also be used, such as moisturizers or agents for improving the natural lipid barrier, such as ceramides, cholesterol sulfates, and / or fatty acids, and mixtures thereof. Enzymes active on the skin may also be used, such as proteases, lipases, cerebrosidases and / or melanases, and mixtures thereof.

[0095] As further examples of active agents that may be suitable for practicing the present invention, there are also the following agents: drugs, peptides, proteins, detectable markers, contrast agents, analgesics, anesthetics, antibacterial agents, antiyeast agents, antifungal agents, antiviral agents, antidermatitis agents, antipruritics, antiemetics, vascular protectants, agents against 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 agents and bronchodilators, sunscreens, antihistamines, healing agents, corticosteroids, tanning agents, and mixtures thereof.

[0096] The content of the at least one active agent in the composition can be adjusted depending on the intended purpose of the composition.

[0097] Adjuvant

[0098] In a known manner, the compositions of the invention may also contain adjuvants customary in cosmetics and / or dermatology, such as preservatives, antioxidants, pH regulators (acidic or basic), fragrances, fillers, bactericides, odor absorbers, colorants (pigments and dyes), emulsifiers, and lipid vesicles.

[0099] Of course, the skilled person will take care to select the optional additional compound(s) and / or the amounts thereof such that the envisaged addition does not or does not substantially adversely affect the benefits of the composition according to the invention.

[0100] Although the numerical ranges and parameters describing the wide range of the present invention are approximate, the numerical values ​​described in the specific examples are reported as accurately as possible. However, any numerical value inherently contains certain errors that are inevitably caused by the standard deviation present in its respective measurements. The following examples are intended to illustrate the present invention without limiting its scope.

[0101] Example

[0102] The amounts / concentrations of the ingredients in the following compositions / formulations are expressed in parts by weight.

[0103] I. Evaluation of the Compositions of the Invention

[0104] 1. In vitro anti-carbonylation efficacy test

[0105] This test provides an in vitro method for determining how anti-carbonylation agents / compositions affect the degree of protein carbonylation. Use deionized water to make a 1% solution of the anti-carbonylation agent / composition to be tested, a 10 mg / mL solution of bovine serum albumin (BSA), 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) are added to the centrifuge tube. The sample containing only BSA and PBS is used as the baseline, and the sample containing BSA, acrolein and PBS is used as the blank. The final liquid volume in each centrifuge tube is 1.0 mL. Each group contains three replicates, and all samples are placed at 37°C overnight.

[0106] Table 1

[0107]

[0108]

[0109] 1.PAMAM comes from Weihai CY Dendrimer Technology Co., Ltd.

[0110] 2. L-Lysine from Macklin.

[0111] 3.Alistin comes from Exsymol.

[0112] The protein carbonyl content in the above samples was detected by using the protein carbonyl detection kit of Nanjing Jiancheng Bioengineering Institute. The detection method is as follows: take 0.1mL of sample and add it to the detection tube, then add 0.4mL of reagent 3 in the kit, vortex mix for 1 minute, and react at 37°C in the dark for 30 minutes. Add 0.5mL of reagent 5 in the kit, vortex mix for 1 minute, centrifuge at 12,000r / min for 10 minutes at 4°C, discard the supernatant, and leave the precipitate. Add 1.0mL of a mixed solution of anhydrous ethanol and ethyl acetate (1:1), vortex mix for 1 minute, centrifuge at 12,000r / min for 10 minutes at 4°C, discard the supernatant, leave the precipitate, and repeat the above steps four times. Add 1.25mL of reagent VI in the kit, mix thoroughly, place it at 37°C for 15 minutes, vortex mix to dissolve all precipitates, and centrifuge at 12,000r / min for 15 minutes. Take the supernatant and add it to a quartz cuvette with a light path of 0.5 cm, adjust it to zero with reagent 6, and measure the absorbance value of each tube at 370nm (ultraviolet). According to the formula, the protein carbonyl content can be obtained.

[0113]

[0114] Micro BCA using Thermos TM Protein detection kit is used to determine the protein content of the sample. First, a diluted albumin (BSA) standard (working range = 20-750 μg / mL) is prepared. BCA reagent A is mixed with reagent B (reagent A: reagent B = 50: 1) to prepare a working solution. 25 μL of each protein standard and protein sample of different dilution concentrations are taken and added to a microplate. 200 μL of working solution is added to each well and fully mixed by shaking on an oscillator for 30 seconds. The microplate is sealed and incubated at 37 ° C for 30 minutes. The microplate is cooled to room temperature and the absorbance of the sample at a wavelength of 562 nm is measured. The corrected absorbance value is the absorbance value of the blank sample subtracted from the reading of each standard and sample. The protein concentration is read against a standard curve prepared with a known amount of BSA. With reference to the standard curve, the protein concentration of each sample is read within the linear range of the standard curve according to the corrected absorbance value of each protein sample. The amount of protein in the original sample is calculated based on the sample volume and dilution.

[0115] 2. Anti-carbonylation effect of PAMAM

[0116] The anti-carbonylation ability of each anti-carbonylation agent / composition can be obtained by dividing the carbonylation degree of the sample by the BSA protein content. The results are shown in Tables 2 and 3 below.

[0117] Table 2

[0118]

[0119] Table 3

[0120]

[0121] 3. Cytotoxicity of PAMAM

[0122] Normal human epidermal keratinocytes (NHEK, FC-0007) purchased from Lifeline Cell Technology were cultured in keratinocyte culture medium (DermaLife K Keratinocyte Medium Complete Kit, LL-0007, Lifeline cell technology) supplemented with growth factors and 1% penicillin / streptomycin (Thermo Fischer, Waltham, MA, USA) at 37°C, 95% humidity and 5% CO2. On the first day, NHEKs were inoculated with culture medium (DermaLife K Keratinocyte Medium Complete Kit) at 3,000 cells / well in 96-well plates. After 24 hours of adherent growth, cells were treated with different types and concentrations of PAMAM for 48 hours. The cells were then washed twice with Dulbecco's phosphate buffered saline (DPBS) and subsequently cultured with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Sigma) for 4 hours. DMSO was then added to dissolve the crystals, followed by measuring the absorbance at 570 nm using a Perkin Elmer plate reader (VICTOR Nivo). The results are shown in Table 4 below.

[0123] Table 4

[0124] Anti-carbonylating agents Safe concentration CYD-100H(PAMAM-OH-G0) ≤1 mg / mL CYD-100A(PAMAM-G0) ≤0.0370mg / mL CYD-110A(PAMAM-G1) ≤0.1111mg / mL CYD-120A(PAMAM-G2) ≤0.0123mg / mL CYD-130A(PAMAM-G3) ≤0.0123mg / mL CYD-140A(PAMAM-G4) ≤0.0041mg / mL Alistin (decarboxylated carnosine) ≤0.0137mg / mL

[0125] II. Formulation Examples

[0126] The PAMAM-containing composition of the present invention is prepared by thoroughly mixing the following ingredients.

[0127] Table 5

[0128]

[0129] III. Sensory Evaluation of Anti-Carbonylation Compositions

[0130] After application of compositions 1 to 4 on the skin, their sensory properties were evaluated and are summarized in the following table:

[0131] Table 6

[0132] Composition Sensory evaluation 1 Good spreadability, good residual softness, medium stickiness 2 Good spreadability, good residual softness, low stickiness 3 Good spreadability, good residual softness, low stickiness 4 Good spreadability, good residual softness, low stickiness

[0133] The cosmetic properties of compositions 1 to 4 were good, especially with respect to spreadability, softness and non-sticky feel.

[0134] Unless expressly excluded or otherwise limited, each document cited herein, including any cross-referenced or related patent or application, is hereby incorporated by reference in its entirety. The citation of any document is not an admission that it is prior art to any invention disclosed or claimed herein, or that it alone or in any combination with any other reference or references teaches, suggests or discloses any such invention. In addition, in the event that any meaning or definition of a term in this document conflicts with any meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to the term in this document shall prevail.

Claims

1. An anti-carbonylation composition comprising at least one anti-carbonylation active substance, wherein the anti-carbonylation active substance comprises at least four and wherein the surface primary amino groups constitute greater than about 8 weight percent of the molecular weight of the anti-carbonylation active.

2. The anti-carbonylation composition according to claim 1, wherein the anti-carbonylation active substance is multi-branched.

3. The anti-carbonylation composition according to claim 1 or 2, wherein the anti-carbonylation active has a molecular weight of about 500 or more, preferably about 800 or more, more preferably about 1000 or more, most preferably about 1200 or more; and / or The anti-carbonylation active has a molecular weight of about 120,000 or less, preferably about 60,000 or less, more preferably about 10,000 or less, most preferably about 5,000 or less; and / or The anti-carbonylation active has a molecular weight of 500 to 120,000, preferably 800 to 60,000, more preferably 1,000 to 10,000, most preferably 1,200 to 5,000.

4. The anti-carbonylation composition according to any one of the preceding claims, wherein the anti-carbonylation active has a structure of formula I, Where R 1 -R 4 Each independently R 5 and R 6 Each independently selected from H and R 7 and R 8 Each independently selected from H and R 9 and R 10 Each independently selected from H and R 11 and R 12 Each independently selected from H and R 13 and R 14 Each independently selected from H and n is selected from 0, 1, 2, 3 and 4; R a and R b Each is independently selected from H, a substituted or unsubstituted C1-C3 alkyl group, wherein the substituted C1-C3 alkyl group contains one or more C1-C3 alkyl substituents, one or more ether bonds, one or more amine groups, or a combination thereof.

5. The anti-carbonylation composition according to claim 4, wherein R 1 -R 4 Each independently R 5 and R 6 Each independently selected from H and R 7 and R 8 Each independently selected from H and R 9 and R 10 Each independently selected from H and n is selected from 0, 1 and 2; R a and R b It's H.

6. The anti-carbonylation composition according to claim 5, wherein the anti-carbonylation active substance is selected from and combinations thereof.

7. The anti-carbonylation composition according to any one of the preceding claims, wherein the anti-carbonylation active is present in the composition in an amount of about 0.0001 wt % to about 10 wt %, preferably about 0.001 wt % to about 5 wt %, preferably about 0.005 wt % to about 2 wt %, relative to the total weight of the anti-carbonylation composition.

8. The anti-carbonylation composition according to any one of the preceding claims, wherein the composition further comprises at least one of a functional agent, a sugar derivative, a natural extract, a vitamin, urea, caffeine, salicylic acid and its derivatives, α-hydroxy acids and their derivatives, retinoids, sunscreens, peppermint essential oil, aloe essential oil or ginseng essential oil, and mixtures thereof.

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

10. A method for preparing the anti-carbonylation composition according to any one of the preceding claims 1 to 8 or the cosmetic according to claim 9, comprising the following steps: - (1) optionally, providing a formulation base, which is then cooled, - (2) dispersing at least one anti-carbonylation active in a cosmetically acceptable vehicle, - (3) mixing 1) the formulation base and 2) the anti-carbonylation active solution at room temperature; Preferably, the formulation base is cooled to room temperature; and / or Preferably, the cosmetically acceptable vehicle is water.

11. A non-therapeutic method for reducing and / or inhibiting protein carbonylation on keratin materials by topically applying the anti-carbonylation composition according to any one of the preceding claims 1 to 8 or the cosmetic according to claim 9 to the keratin materials; Preferably, the keratin material is skin.

12. A non-therapeutic method for protecting keratin materials from carbonylation stimulation, which is carried out by topically applying the anti-carbonylation composition according to any one of the preceding claims 1 to 8 or the cosmetic according to claim 9 to the keratin materials; Preferably, the keratin material is skin.

13. Non-therapeutic use of an anti-carbonylation composition according to any one of the preceding claims 1 to 8 or a cosmetic according to claim 9 for reducing and / or inhibiting protein carbonylation on keratin materials; Preferably, the keratin material is skin.

14. Non-therapeutic use of an anti-carbonylation composition according to any one of the preceding claims 1 to 8 or a cosmetic according to claim 9 for protecting keratin materials from carbonylation irritation; Preferably, the keratin material is skin.

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