Azuki bean protein hydrolysates and their cosmetic uses

Abstract

The present invention relates to a protein hydrolysate obtained from adzuki beans and its uses, in particular in the cosmetic treatment and / or care of the skin and / or hair. The protein hydrolysate is selected from the group consisting of (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2), (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv) Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4). Also disclosed is a method for preparing the protein hydrolysate.

Description

[Technical field]

[0001] The present invention relates to protein hydrolysates obtained from adzuki beans and their uses, in particular their use in cosmetic treatment and / or care of skin and / or hair. The protein hydrolysates are useful in protecting skin and / or hair from the harmful effects of exposomes, including those resulting from air pollution and / or sunlight. Furthermore, the protein hydrolysates are useful in protecting hair from chemical and thermal stress, such as those resulting from personal care regimes. Also disclosed is a method for preparing the protein hydrolysates. [Background technology]

[0002] The totality of extrinsic factors to which an individual is exposed throughout his or her life is referred to as the exposome. Skin appearance, hair appearance, and aging are influenced by the combined action of intrinsic and extrinsic factors. Intrinsic factors that negatively affect skin and hair include, for example, chronological aging, the genetic makeup of humans, and other biological changes that occur from within the skin. For decades, the damaging extrinsic factors to skin and hair have been ignored. However, there is growing evidence that exposomes may have adverse effects on skin and hair appearance [Vierkotter A1, et al. Airborne particle exposure and extrinsic skin aging. J Invest Dermatol. 2010, and Paarado C et al "Environmental Stressors on Skin Aging. Mechanistic insights", Front Pharmacol. 2019;10:759, published July 9, 2019, doi:10.3389 / fphar.2019.00759].

[0003] Examples of exogenous factors associated with the exposome include ultraviolet (UV) radiation emitted by the sun, high-energy visible light (blue-violet) emitted by devices such as TV / computer / smartphone screens, and exposure to harmful chemical agents found in air pollution, such as smog and cigarette smoke.

[0004] Ultraviolet (UV) radiation is one of the main extrinsic factors that have a deleterious effect on the appearance of skin and hair. Extrinsic aging is believed to be primarily the result of exposure to solar radiation and is also known as photoaging.

[0005] After thorough studies of the effects of pollution on health, there is an increasing awareness of the harmful effects of pollution on the appearance of skin and hair [Vierkotter A, et al. Airborne particle exposure and extrinsic skin aging. J Invest Dermatol. 2010]. In addition, it is known that the harmful effects of various pollutants on skin and hair can be amplified due to their interaction with UV radiation [Drakaki E, et al. Air pollution and the skin. Front. Environ. Sci., 2014]. For example, it is well known that the main role of the skin is to form a barrier between the host and the environment. The skin must protect the organism from chemicals, ultraviolet light, mechanical damage, and pathogenic microorganisms. Also, the skin must provide an efficient permeability barrier that prevents the loss of water and electrolytes. [Feingold K, Elias P. The important role of lipids in the epidermis and their role in the formation and maintenance of the cutaneous barrier. Biochim Biophys Acta. 2014]. Air pollutants may have a major effect on skin function. Changes that disrupt the skin barrier function, either in the stratum corneum lipid metabolism or in the protein components of the keratinocytes, adversely affect skin appearance. Air pollutants may induce severe interference with the normal function of lipids, DNA, and / or proteins in human skin through oxidative damage [Eleni Drakaki, Clio Dessinioti and Christina V. Antoniou. Air pollution and the skin. Front. Environ. Sci., 2014].

[0006] Moreover, continuous anthropogenic emissions of greenhouse gases (mainly carbon dioxide) have led to an increase in the Earth's temperature of more than 1°C since 1880, with most of the warming occurring in the past 45 years [National Aeronautics and Space Administration. NASA ozone watch. 2020b [cited 4 April 2020]. Available at: https: / / ozonewatch.gsfc.nasa.gov / facts / history_SH.html]. Thus, solar radiation and pollutants are two important components of the exposome, but also other environmental components, such as greenhouse gases and temperature (which affect all biochemical reactions), are highly relevant.

[0007] The above exogenous factors can result in high production of reactive oxidative species (ROS). In skin, production of ROS can affect skin defenses by rapidly depleting enzymatic (glutathione peroxidase, glutathione reductase, superoxide dismutase, catalase) and nonenzymatic (vitamin E, vitamin C, and glutathione) antioxidant capacity [Valacchi G, et al.Cutaneous responses to environmental stressors.Ann NY Acad Sci.2012]. In hair, oxidative stress can also have significant negative effects. Proteins are the main components (65%-95%) of the hair shaft, and protein carbonylation is an irreversible oxidative protein modification that can significantly damage hair. Protection of hair proteins from carbonylation caused by pollutants is desirable for hair and scalp protection from urban stress. [Baraibar, Martin. Urban pollution induces irreversible carbonylation of hair proteins. Conference: IFSCC, 2018].

[0008] In addition, the skin, and especially the hair, are also subjected to other stressors from daily personal care routines. The various causes of extrinsic damage to the hair shaft can be broadly divided into physical and chemical causes. The human hair shaft is composed of a cortex with a central shaft medulla and an outer stratum corneum. Chemical causes of hair damage include bleaching and hair dyeing, while physical causes include heat treatment (e.g., treatment with a straightening iron), friction from hair accessories, and washing [Lee Y, et al. Comparison of hair shaft damage after chemical treatment in Asian, White European, and African hair. Int J Dermatol. 2014].

[0009] The effects of extrinsic factors on skin and hair can be differentiated from those associated with intrinsic factors. Skin adversely affected by the exposome can exhibit more symptoms of aging, such as coarse wrinkles. Hair adversely affected by the exposome can have a damaged appearance and can be more difficult to manage.

[0010] From a preventative standpoint, it is of particular interest to counter exposome factors (ie, exogenous factors) since exogenous factors can be more easily altered than endogenous factors.

[0011] It is desirable for skin and / or hair to have an aesthetically pleasing appearance.There is a great need to find active ingredients that can protect skin, scalp and hair from exposomal attacks, such as environmental attacks that they may be exposed to daily, including stressors that result from daily personal care routines.In particular, there is a great need to find such active ingredients that can provide such protection for the aesthetic benefit of skin and / or hair.

[0012] US Patent Publication No. 20210177733 discloses compositions and methods for enhancing the ability of the skin to defend itself against free radical attack caused by exposome-induced oxidative stress. The compositions include an Aristotelia chilensis leaf extract, a Buddleja globosa leaf extract, and an Ugni molinae leaf extract.

[0013] Zoryalys® from Greentech is a natural active ingredient "derived from several molecules: shogaol and gingerol (which grow in sunny areas), magnolol and honokiol (which live in polluted areas), and prakashi (a biomimetic lipid)" (https: / / www.greentech.fr / en / zoryalys-2 / ). ZORYALYS® is said to protect hair from protein oxidation and is active from the surface to the core of the hair fiber, allowing complete protection of hair from exposome attacks. The active is said to maintain hair mechanical properties and improve hair shine by maintaining keratin integrity for healthy, beautiful hair.

[0014] Peptides released from extruded adzuki bean protein through simulated gastrointestinal digestion exhibit anti-flammatory activity are disclosed in Zhensing Shi et al, "Peptides released from extruded adzuki bean protein through simulated gastrointestinal digestion exhibit anti-flammatory activity," J. Agric. Food Chem. 2021, 69, 7028-7036. There remains a need for alternative plant-based active compositions that can protect skin and / or hair from exposome-induced oxidative stress. There remains a need for alternative plant-based active compositions that can protect / maintain / improve the aesthetic properties of skin and / or hair when exposed to exposomes. The present invention aims to meet some or all of these needs and to solve some or all of the problems identified above. [Prior art documents] [Patent documents]

[0015] [Patent Document 1] US Patent Publication No. 20210177733 [Non-patent literature]

[0016] [Non-Patent Document 1] Vierkotter A1,et al.Airborne particle exposure and extrinsic skin aging.J Invest Dermatol.2010 [Non-Patent Document 2] Paarado C et al "Environmental Stressors on Skin Aging. Mechanistic insights", Front Pharmacol.2019;10:759, published July 9, 2019, doi:10.3389 / fphar.2019.00759 [Non-Patent Document 3] Drakaki E,et al.Air pollution and the skin.Front.Environ.Sci.,2014 [Non-Patent Document 4] Feingold K,Elias P.The important role of lipids in the epidermis and their role in the formation and maintenance of the cutaneous barrier.Biochim Biophys Acta.2014 [Non-Patent Document 5] Eleni Drakaki,Clio Dessinioti and Christina V.Antoniou.Air pollution and the skin.Front.Environ.Sci.,2014 [Non-Patent Document 6] National Aeronautics and Space Administration.NASA ozone watch.2020b [Non-Patent Document 7] Valacchi G,et al.Cutaneous responses to environmental stressors.Ann NY Acad Sci.2012 [Non-Patent Document 8] Baraibar, Martin.Urban pollution induces irreversible carbonylation of hair proteins.Conference:IFSCC,2018 [Non-Patent Document 9] Lee Y,et al.Comparison of hair shaft damage after chemical treatment in Asian,White European,and African hair.Int J Dermatol.2014 [Non-Patent Document 10] https: / / www.greentech.fr / en / zoryalys-2 / [Non-Patent Document 11] Zhensing Shi et al, "Peptides released from extruded adzuki bean protein through simulated gastrointestinal digestion exhibit anti-flammatory activity", J.Agric.Food Chem.2021,69,7028-7036 Summary of the Invention

[0017] In a first aspect, the present invention provides a method for producing a composition comprising the steps of: (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4). The protein hydrolysate additionally contains the following compounds: (v) Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (sequence number 5).

[0018] The protein hydrolysates of the present invention have been found to be useful in reducing or preventing the adverse effects of exposomes on skin and / or hair. Adverse effects on skin and / or hair include those resulting from air pollution and / or sunlight. Adverse effects on hair also include those resulting from chemical treatments, such as dyeing, and physical treatments, such as styling aids, such as hair dryers or straighteners. Adverse effects on hair may manifest in the form of damaged hair, more unmanageable hair, hair with more split ends, hair with reduced fiber alignment, hair with increased frizz, and / or hair with less hydration. Adverse effects on skin may manifest in the form of a reduction in barrier function, a dull appearance of the skin, loss of skin brightness, luminosity, and / or luster, loss of homogeneity in skin texture, an increase in skin roughness, the appearance of dark spots, and / or erythema (skin redness).

[0019] In another aspect, the present invention provides a method for producing a pharmaceutical composition comprising: (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and The present invention relates to its stereoisomers and cosmetically acceptable salts.Surprisingly, these compounds are found to exhibit excellent antioxidant activity.Without wishing to be bound by theory, it is believed that this excellent antioxidant activity of these compounds contributes to the ability of protein hydrolysate to have the performance described herein.

[0020] In one aspect, the present invention relates to a combination of a protein hydrolysate according to the first aspect of the invention and a filtrate obtained by fermentation of Raphanus sativus roots with the microorganism Leuconostoc. This combination has been found to produce a synergistic effect in terms of antioxidant activity. The present invention also relates to (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and It extends to a combination of its stereoisomers and cosmetically acceptable salts with a filtrate obtained by fermentation of the roots of Raphanus sativus by the microorganism Leuconostoc.

[0021] In one aspect, the present invention provides a process for preparing azuki bean protein hydrolysate, the hydrolysate comprising: a) providing a suspension of ground azuki beans in water; b) treating the suspension with alkali to solubilize proteins in the ground adzuki beans; c) enzymatically treating the suspension obtained in b) with a serine endoprotease; d) inactivating serine endoproteases in the suspension obtained in c) to form a crude hydrolysate; e) recovering the supernatant from the crude hydrolysate by solid / liquid separation; f) removing components having a molecular weight of more than 3 KDa from the supernatant.

[0022] The process is useful in the preparation of a protein hydrolysate according to the first aspect of the invention.The invention extends to a protein hydrolysate obtainable by the process for preparing an azuki bean protein hydrolysate described herein.

[0023] In one aspect, the present invention provides a method for the preparation of a protein hydrolysate of adzuki bean according to the first aspect of the invention, comprising the steps of: (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and The present invention relates to a personal care or cosmetic composition comprising said stereoisomers and cosmetically acceptable salts thereof, or a combination of said compounds with a filtrate obtained by fermentation of the roots of Raphanus sativus by the microorganism Leuconostoc.

[0024] In one aspect, the present invention relates to an azuki bean protein hydrolysate according to the first aspect of the invention for the treatment and / or care of the skin and / or hair, or (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and The present invention relates to the use of its stereoisomers and cosmetically acceptable salts, or a combination of said compounds with a filtrate obtained by fermentation of the roots of Raphanus sativus by the microorganism Leuconostoc, in particular for the treatment and / or care of the skin and / or hair, both cosmetic and non-therapeutic.

[0025] In one aspect, the present invention relates to a method of treatment and / or care of the skin and / or hair, comprising administering to a subject an azuki bean protein hydrolysate according to the first aspect of the invention, or (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and The present invention relates to a method comprising administering a combination of said compounds and a filtrate obtained by fermentation of the roots of Raphanus sativus with the microorganism Leuconostoc, and / or a combination of said compounds and a filtrate obtained by fermentation of the roots of Raphanus sativus with the microorganism Leuconostoc. Treatment and / or care includes reducing or preventing the harmful effects of exposomes on the skin and / or hair, reducing or preventing the harmful effects of air pollution on the skin and / or hair, reducing or preventing the harmful effects of sunlight on the skin and / or hair, reducing or preventing the harmful effects of chemical treatments of hair, reducing or preventing the harmful effects of heat stress on hair, and / or maintaining and / or improving the physical barrier function of the skin. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0026] The present invention is based on the discovery of surprising properties of azuki bean protein hydrolysates, which make them suitable for non-therapeutic cosmetic applications on the skin and / or hair.

[0027] definition In the context of the present invention, "skin" is understood to mean the layers that make up the skin, including both the top layer or stratum corneum to the bottom layer or subcutaneous tissue. These layers are composed of different types of cells, such as keratinocytes, fibroblasts, melanocytes, and / or adipocytes, among others. In the context of the present invention, the term "skin" includes the scalp. The term "skin" includes mammalian skin, such as human skin, including skin containing hair.

[0028] In the context of the present invention, "hair" includes hair on the scalp, skin, eyelashes, eyebrows, moustache region, and / or beard region of a subject.

[0029] As used in the context of this specification, the term "treatment" when accompanied by the condition "cosmetic" means that the treatment is non-therapeutic and has the purpose of improving the aesthetic appearance of the skin and / or hair, including the improvement of the skin and / or hair properties that affect the cosmetic appearance of the skin and / or hair, such as, but not limited to, the level of hydration, elasticity, firmness, shine, tone, or texture. The term "care" in the context of this specification refers to the maintenance of the skin and / or hair properties. The skin and / or hair properties are subject to improvement and maintenance through cosmetic treatment and / or care, both in healthy subjects and in subjects presenting with skin and / or hair diseases and / or disorders, such as, but not limited to, ulcers and lesions on the skin, psoriasis, dermatitis, acne or rosacea, alopecia, among others.

[0030] As used herein, the term "prevent" refers to the ability of the actives of the present invention to prevent, delay, or impede the appearance or development of symptoms or signs of adverse effects on the skin and / or hair. As used herein, the term "ameliorate" refers to the ability of the actives of the present invention to reduce or lessen symptoms or signs of adverse effects on the skin and / or hair.

[0031] As used herein, the term "about," for example when referring to a measurable value (e.g., an amount or weight of a particular ingredient, or temperature), refers to a variation of ±20%, ±10%, ±5%, ±1%, ±0.5%, or especially ±0.1% of the specified amount.

[0032] As used herein, the term "comprising" is inclusive or open-ended and does not exclude additional, unrecited elements or method steps, and is intended to encompass the phrases "consisting essentially of" and "consisting of" as alternative embodiments, where "consisting of" excludes any elements or steps not specified, and "consisting essentially of" permits the inclusion of additional, unrecited elements or steps that do not materially affect the essential or basic and novel characteristics of the composition or method under consideration.

[0033] The abbreviations used for amino acids herein follow the rules of the IUPAC-IUB Commission of Biochemical Nomenclature as set forth in Eur.J.Biochem.,(1984),138,9-37. Thus, for example, Gly represents NH2-CH2-COOH, Gly- represents NH2-CH2-CO-, -Gly represents -NH-CH2-COOH, -Gly- represents -NH-CH2-CO-. Thus, the hyphen representing a peptide bond eliminates the OH in the 1-carboxyl group of an amino acid (here represented in the conventional non-ionized form) when located to the right of the symbol, and eliminates the H in the 2-amino group of an amino acid when located to the left of the symbol, and both modifications can be applied to the same symbol (see table below).

[0034] [Table 32]

[0035] Azuki bean protein hydrolysate and its preparation process In a first aspect, the present invention relates to an azuki bean protein hydrolysate.

[0036] Adzuki bean, also known as Vigna angularis, is a legume originally cultivated in the highlands near China and Tibet and consumed worldwide as a rich source of protein (over 20% percent). Vigna angularis is also known as adzuki bean (Japanese: azuki, azuki, adzuki), azuki bean, or red mung bean.

[0037] A protein hydrolysate is a mixture of fragments resulting from the hydrolysis of a protein, including, for example, free amino acids and peptides of different molecular weights and compositions. The protein hydrolysate of the present invention may be obtained from the enzymatic hydrolysis of azuki bean proteins.

[0038] The protein hydrolysate is (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4).

[0039] Thus, protein hydrolysates are (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), and (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (sequence number 2).

[0040] The protein hydrolysate is (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv) Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn (sequence number 4).

[0041] The protein hydrolysate is (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4).

[0042] The protein hydrolysate is (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4).

[0043] The protein hydrolysate is (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4).

[0044] Protein hydrolysates, particularly protein hydrolysates having any of the combinations of compounds labeled (i), (ii), (iii), or (iv) above, may additionally contain the following compounds: (v) Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (sequence number 5).

[0045] Each of the compounds labeled (i)-(v) above has been found to have excellent antioxidant activity. Without wishing to be bound by theory, it is believed that this activity contributes to the advantageous skin / hair treatment and / or care properties exhibited by the protein hydrolysates of the invention.

[0046] When present in the protein hydrolysate, each of these compounds labeled (i)-(v) above is present in the protein hydrolysate in an amount of 0.001-0.15 ppm or 0.005-0.08 ppm. When present in the protein hydrolysate, the compound labeled (i) above can be present in the protein hydrolysate in an amount of 0.001-0.1 ppm, 0.005-0.05 ppm, or 0.01-0.03 ppm. When present in the protein hydrolysate, the compound labeled (ii) above can be present in the protein hydrolysate in an amount of 0.001-0.1 ppm, 0.005-0.05 ppm, or 0.015-0.04 ppm. When present in the protein hydrolysate, the compound labeled (iii) above can be present in the protein hydrolysate in an amount of 0.001-0.1 ppm, 0.005-0.05 ppm, or 0.01-0.03 ppm. When present in the protein hydrolysate, the compound labeled (iv) above can be present in the protein hydrolysate in an amount of 0.005-0.15 ppm, 0.01-0.1 ppm, or 0.03-0.08 ppm. When present in the protein hydrolysate, the compound labeled (v) above can be present in the protein hydrolysate in an amount of 0.001-0.15 ppm, 0.001-0.01 ppm, or 0.003-0.008 ppm.

[0047] The protein hydrolysate can have a molecular weight of 3 kDa or less.Accordingly, the protein hydrolysate can be devoid of any component with a molecular weight of more than 3 kDa, i.e., does not contain any component with a molecular weight of more than 3 kDa.This can be achieved by filtering the hydrolysate.

[0048] The protein hydrolysate can be obtained by treating a suspension of ground azuki beans in water with alkali to solubilize the proteins in the ground azuki beans, treating the resulting suspension with serine endoprotease, inactivating the serine endoprotease in the suspension to form a crude hydrolysate, and purifying the crude hydrolysate. In this case, the enzymatic hydrolysis of the proteins in the ground azuki beans is performed in the suspension (the suspension of ground azuki beans in water in which the proteins in the azuki beans are solubilized) and not on the isolated proteins in any form. Purifying can include removing any components having a molecular weight of more than 3 kDa from the crude hydrolysate. The protein hydrolysate of the present invention is the purified product of this process. The process may not include a step of isolating proteins from the ground azuki beans and a step of subjecting the isolated proteins to enzymatic hydrolysis. The process may not include a step of subjecting the proteins isolated from the azuki beans to enzymatic hydrolysis, and as a result, the protein hydrolysate contains components other than protein fragments, such as polyphenols and other organic compounds.

[0049] In particular, the protein hydrolysate may contain polyphenols in an amount of at least 0.05 mg polyphenols per milliliter of protein hydrolysate, in particular at least 0.15 mg polyphenols per milliliter of protein hydrolysate. In particular, the amount of polyphenols may range from 0.05 to 0.30 mg / mL, more in particular from 0.15 to 0.25 mg / mL. The amount of polyphenols may be measured by the Folin-Ciocalteu method, in particular expressed as mg gallic acid equivalents per mL of protein hydrolysate. The Folin-Ciocalteu method is a method well known to those skilled in the art in the analysis of compositions derived from botanicals and is widely described in the literature (e.g. Lamuela-Raventos, "Folin-Ciocalteu method for the measurement of total phenolic content and antioxidant capacity" 2018, Measurement of Antioxidant Activity & Capacity: Recent Trends and Applications). Alternatively, the protein hydrolysate may contain polyphenols in an amount of at least 2 mg polyphenols per g protein hydrolysate (dry weight), in particular at least 6 mg / g. In particular, the amount of polyphenols may range from 2 to 12 mg / g, more in particular from 6 to 9 mg / g. By polyphenols is meant naturally occurring organic compounds characterized by the presence of multiple phenolic units. Polyphenols include, for example, flavonoids, tannic acid, and ellagitannins.

[0050] The protein hydrolysate may have a dry weight of 2-5 mg, a protein content of 19-35 w / w%, a DPPH activity of 74-80%, and a polyphenol content of 0.05-0.30 mg / mL or 2-12 mg / g.

[0051] The protein hydrolysate may be prepared by the following steps a) to d) to obtain a crude hydrolysate, followed by purification of the crude hydrolysate.

[0052] Step a) involves providing a suspension of ground azuki beans in water. The azuki beans are raw azuki beans. The ground azuki beans may be obtained by grinding the azuki beans into a fine flour. The flour may be sieved to remove larger fragments. Typically, the ground azuki beans have a particle size of less than 250 micrometers.

[0053] Step b) involves treating the suspension of ground azuki beans in water with an alkali to solubilize the proteins of the ground azuki beans in water. The purpose of this step is to increase the content of solubilized proteins in the suspension. Typically, this step involves adjusting the pH of the suspension to a pH of about 6.5 to about 7.5, e.g., a pH of 7. The pH of the suspension affects the amount and type of proteins in the suspension. This can be achieved by adding an alkali, e.g., NaOH, to the suspension of ground azuki beans in water. Typically, step b) is carried out at room temperature, i.e., 22±2° C. After the alkali is added to the suspension of ground azuki beans, the suspension is kept at room temperature for a period of time, typically 1 hour, before step c).

[0054] Step c) comprises enzymatically treating the suspension obtained in step b) with a serine endoprotease (also known as serine endopeptidase). The serine endoprotease can be subtilisin A. Subtilisin A (classification EC 3.4.21.62, peptidase family S8) is also known as subtilisin Carlsberg, subtilopeptidase A, Alcalase®, ALK enzyme, bacillopeptidase A, Bacillus subtilis alkaline protease bioprase, bioprase AL, colistinase, genenase I, Esperase®, maxatase, protease XXVII, thermoase, superase, subtilisin DY, subtilopeptidase, SP 266, Savinase®, kazusase, protease VIII, protin A 3L, orientase 10B, or protease S. Subtilisin A is commonly obtained from Bacillus licheniformis. Step c) is carried out at a temperature of 20-30°C, and may be carried out at a temperature of 23-27°C or at a temperature of about 25°C. Step c) may be carried out for a period of, for example, about 30-120, about 45-90, or about 60 minutes. Typically, the enzyme activity per 1 g of protein in the ground azuki beans is 0.1-0.5 AU / g. AU stands for Anson Unit. Anson Unit is a unit of enzyme concentration defined as the amount of enzyme that can digest urea-denatured hemoglobin at the same initial rate as 1 milliequivalent of tyrosine under standard conditions (25C and pH of 7.50). Typically, the percentage of protein in the ground azuki beans is 15-25% w / w. Typically, the concentration of ground azuki beans in water is 20-100 g (ground azuki beans) / Kg (water).

[0055] Step d) involves inactivating the enzymes to obtain a crude isolate. During step c), the pH of the suspension (subjected to enzymatic hydrolysis) is monitored. A decrease in pH is associated with the release of peptide fragments and the subsequent accumulation of acidic carboxylic acids. Typically, during step c), the suspension subjected to enzymatic hydrolysis undergoes a slight acidification, which results in a decrease in pH of, for example, 0.2 to 0.3 units. After the enzymatic hydrolysis step c), the enzymes are inactivated by increasing the temperature of the suspension obtained in c) to at least 70°C, or 70 to 90°C, or 75 to 85°C. The suspension obtained in step c) may be obtained at this temperature for a period of time, preferably at least about 20 minutes. The product of step d) is a crude hydrolysate.

[0056] The crude hydrolysis product of step d) is then subjected to purification to remove any components with a molecular weight of more than 3 kDa. This can be achieved by solid / liquid separation (step e)) and ultrafiltration (step f)). Step e) is a purification step and includes removing solid materials from the crude hydrolysate by solid / liquid separation. Suitable separation techniques are known in the art and include, for example, filtration, sedimentation, decantation, and / or centrifugation. Filtration can be carried out using filters with a pore size of less than 1000 μm, or less than 20 μm, or less than 10 μm, or less than 1 μm, or less than 0.1 μm. Filtration can be carried out in successive filtration operations, for example using multiple filters with decreasing pore size. Step f) is also a purification step and includes ultrafiltration to remove fragments with a molecular weight of more than 3 kDa from the supernatant obtained in step e). Ultrafiltration is a technique well known in the art and involves the use of membranes to separate molecules with different molecular weights, in this case a cut-off threshold of 3 kDa.

[0057] The present invention extends to a protein hydrolysate obtainable by the above process.

[0058] The present invention relates to (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), and (v) Compounds selected from: Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5). Each of the compounds labeled (i)-(v) above has been found to have excellent antioxidant activity, and without wishing to be bound by theory, it is believed that this activity contributes to the advantageous skin / hair treatment and / or care properties exhibited by the protein hydrolysates of the invention.

[0059] The present invention also extends to the combination of a protein hydrolysate according to the first aspect of the invention and a filtrate obtained by fermentation of Raphanus sativus roots with the microorganism Leuconostoc.

[0060] The present invention relates to (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and It extends to a combination of its stereoisomers and cosmetically acceptable salts with a filtrate obtained by fermentation of the roots of Raphanus sativus by the microorganism Leuconostoc.

[0061] This filtrate obtained by fermentation of Raphanus sativus roots with the microorganism Leuconostoc contains antimicrobial peptides and acts as a preservative. One example of a commercially available filtrate obtained by fermentation of Raphanus sativus roots with the microorganism Leuconostoc is Leucidal® (INCI name: Leuconostoc / radish root fermentation filtrate) from Active Micro Technologies LCC. The most preferred species of Leuconostoc is Leuconostoc kimchi. Leucidal® is typically used as a preservative. It has been found that the antioxidant activity of the protein hydrolysate of the present invention is unexpectedly enhanced in the presence of this filtrate.

[0062] composition The present invention includes a composition, which may be a personal care composition or a cosmetic composition, comprising a protein hydrolysate as described herein, together with at least one cosmetically acceptable excipient or adjuvant. The present invention extends to a composition, which may be a personal care composition or a cosmetic composition, comprising a combination of a protein hydrolysate as described herein and a filtrate obtained by fermentation of Raphanus sativus root with the microorganism Leuconostoc ... (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and The present invention extends to compositions comprising the stereoisomers and cosmetically acceptable salts thereof, together with at least one cosmetically acceptable excipient or adjuvant. The present invention relates to a composition, which may be a personal care composition or a cosmetic composition, comprising: (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and The present invention extends to compositions comprising a combination of stereoisomers and cosmetically acceptable salts thereof and a filtrate obtained by fermentation of the roots of Raphanus sativus by the microorganism Leuconostoc, together with at least one cosmetically acceptable excipient or adjuvant.

[0063] These compositions may be prepared by conventional means known to those skilled in the art ["Harry's Cosmeticology", Seventh edition, (1982), Wilkinson JB, Moore RJ, eds. Longman House, Essex, GB].

[0064] The composition comprises a cosmetically effective amount of the protein hydrolysate to be administered, or (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and The dosage of the compound, including its stereoisomers and cosmetically acceptable salts, will likewise depend on a number of factors, including the age, condition of the patient, the nature or severity of the condition, the disorder or disease being treated and / or cared for, the route and frequency of administration, and the specific nature of the compound used.

[0065] The term "cosmetically effective amount" is understood to mean a non-toxic but sufficient amount of the extract of the present invention to provide the desired effect. The protein hydrolysate may be present in an amount of at least 0.001% by weight, in particular about 0.001-10% by weight, preferably about 0.004-5% by weight, based on the total weight of the personal care or cosmetic composition.

[0066] The composition can be a composition for topical application, which optionally comprises cosmetically acceptable excipients necessary to formulate a desired dosage form. A topical composition is a composition suitable for topical application to mammalian keratinous tissue, such as the skin or hair.

[0067] Compositions for topical or transdermal application can be made into any solid, liquid, or semi-solid formulation, for example, but not limited to, creams, multiple emulsions, including but not limited to, oil-in-water and / or silicone-in-water emulsions, water-in-oil and / or water-in-silicone emulsions, water / oil / water or water / silicone / water emulsions, and oil / water / oil or silicone / water / silicone emulsions, anhydrous compositions, aqueous dispersions, oils, milks, balsams, foams, lotions, gels, cream gels, hydroalcoholic solutions, hydroglycolic solutions, hydrogels, liniments, serums, soaps, shampoos, conditioners, serums, polysaccharide films, ointments, mousses, pomades, powders, bars, pencils, and sprays or aerosols (sprays), including leave-on and rinse-off formulations. These topical or transdermal formulations can be incorporated using techniques known to those skilled in the art into various types of solid equipment, including, but not limited to, bandages, gauze, T-shirts, socks, tights, underwear, girdles, gloves, diapers, sanitary napkins, bandages, bed covers, wipes, adhesive patches, non-adhesive patches, occlusive patches, micro-electrical patches or face masks, or into various makeup products, such as makeup foundations, including liquid foundations and compact foundations, makeup remover lotions, makeup remover emulsions, dark circle concealers, eye shadows, lipsticks, lip protectors, lip glosses and powders, among others.

[0068] The compositions of the present invention may be in a form selected from a cream, a multiple emulsion, a solution, a liquid crystal, anhydrous composition, an aqueous dispersion, an oil, an emulsion, a balsam, a foam, a lotion, a gel, a cream gel, a hydroalcoholic solution, a hydroglycolic solution, a hydrogel, a liniment, a soap, a shampoo, a conditioner, a serum, a polysaccharide film, an ointment, a mousse, a pomade, a powder, a bar, a pencil, a spray, or an aerosol.

[0069] The compositions of the invention may include an agent that increases the transdermal absorption of the compounds of the invention, such as, but not limited to, dimethylsulfoxide, dimethylacetamide, dimethylformamide, a surfactant, azone (1-dodecylazacycloheptan-2-one), alcohol, urea, ethoxydiglycol, acetone, propylene glycol, or polyethylene glycol, among others.

[0070] In particular, the compositions are for topical application and are hair care compositions, such as conditioners, treatments, hair tonics, styling gels, mousses, shampoos, hairsprays, pomades, setting lotions, colorings, and permanent wave compositions. Particularly relevant for the purposes of the present invention are tonics, conditioners, treatments, and styling gels, which may be in the form of gels, lotions, tinctures, sprays, mousses, cleansing compositions, or foams, and may be applied, depending on the individual needs, for example, once a day as a lotion, tincture, mousse, or spray, or once or twice a week as a conditioner or treatment. In particular, suitable compositions in the context of the present invention are lotions, shampoos, serums, or eyelash mascaras. In particular, the cosmetic composition is a serum. The term "serum" is well known to those skilled in the art and refers to a composition that is transparent, gel-based, or liquid. Serums have a fluid texture and are more concentrated in active agents than standard care products.

[0071] In addition, the composition may contain other active ingredients, such as anti-wrinkle agents, botox-like agents, and / or anti-aging agents; firming agents, skin elasticity agents, and / or restructuring agents; moisturizing agents; anti-photoaging agents and / or blue light protection agents; DNA protection agents, DNA repair agents, and / or stem cell protection agents; free radical scavengers, and / or anti-glycation agents, detoxifying agents, antioxidants, and / or anti-pollution agents; antiperspirants; melanin synthesis stimulators or inhibitors; whitening or depigmenting agents; pre-pigmenting agents; self-tanning agents; lipolytic agents or lipolytic stimulators, adipogenic agents. Additional examples can be found in CTFA International Cosmetic Ingredient Dictionary & Handbook, 12th Edition (2008).

[0072] Anti-wrinkle, botox-like, and / or anti-aging agents include Oxygestkin® [INCI: Tropaeolum majus flower / leaf / stem extract] or Eclaline™ from SILAB [INCI: Hydrolyzed Lupin Protein Octenyl Succinate]; Matrixyl® [INCI: Palmitoyl Pentapeptide-4], Matrixyl® 3000® [INCI: Palmitoyl Tetrapeptide-7, Palmitoyl Oligopeptide], Matrixyl® Synthe'6 [INCI: Glycerin, Water, Hydroxypropyl Cyclodextrin, Palmitoyl Tripeptide-38 ... Morphomics™ [INCI: Pentylene Glycol, Caprylyl Glycol], Essenskin™ [INCI: Calcium Hydroxymethionine], Renovage [INCI: Teprenone], Dermaxyl® [INCI: Palmitoyl Oligopeptide], Calmosensine [INCI: Butylene Glycol, Acetyl Dipeptide-1 Cetyl Ester], Volulip [INCI: Cetearyl Ethylhexanoate, Sorbitan Isostearate, Portulaca Pilosa extract, sucrose coconut, palmitoyl tripeptide-38], Subliskin [INCI: Sinorhizobium meliloti ferment, cetyl hydroxyethylcellulose, lecithin], Biopeptide CL [INCI: palmitoyl oligopeptide], Biopeptide EL [INCI: palmitoyl oligopeptide], Rigin [INCI: palmitoyl tetrapeptide-3], Biobustyl [INCI: glyceryl polymethacrylate, Rahnella / soy protein ferment, palmitoyl oligopeptide], Dynalift [INCI: sodium polystyrene sulfonate, Sorghum bicolor stalk juice, glycerin], Idealift [INCI: acetyl dipeptide-1 cetyl ester], Siegesbeckia [INCI: SiegesbeckiaOrientales extract], Ovaliss [INCI: Coco-glucoside, Caprylyl glycol, Alcohol, Glaucine], Juvinity™ [INCI: Geranylgerani-isopropanol], Prolevis [INCI: Hydrolyzed Vegetable Protein], Idealift™ [INCI: Hydroxyethylcellulose, Acetyl Dipeptide-1 Cetyl Ester], Beautifeye™ [INCI: Albizia Julibrissin peel extract, Darutoside], Chromocare™ [INCI: Sigesbeckia Orientalis extract, Rabdosia Rubescens extract] or Resistem [suggested INCI: Globularia Cordifolia ferment]. Vialox® [INCI: Pentapeptide-3], Syn®-Ake® [INCI: Dipeptide diaminobutyroyl benzylamide diacetate], Syn®-Coll [INCI: Palmitoyl tripeptide-5], Phytaluronate [INCI: Ceratonia Siliqua (carob) gum], Preregen® [INCI: Glycine], soja (soybean) protein, oxidoreductase enzymes], Pepha-Nutrix [INCI: Natural nutritional factors], Pepha-Tight [INCI: Algae extract, pullulan], Pentacare-NA [INCI: Hydrolyzed wheat gluten, carob gum], Syn®-Tacks [INCI: Glycerin, Palmitoyl Dipeptide-5 Diaminobutyroyl Hydroxythreonine, Palmitoyl Dipeptide-6 Diaminohydroxybutyrate], BeauActiveMTP [INCI: Hydrolyzed Milk Protein], Syn®-TC [INCI: Tetradecylaminobutyroylvalylaminobutyric Urea Trifluoroacetate, Palmitoyl Tripeptide-5, Palmitoyl Dipeptide-5 Diaminobutyroyl Hydroxythreonine], Syn®-Hycan [INCI: Tetradecylaminobutyroylvalylaminobutyric Urea Trifluoroacetate], Syn®-Glycan [INCI: Tetradecylaminobutyroylvalylaminobutyric Urea Trifluoroacetate], Regu-Age [INCI: Hydrolyzed Rice Bran Protein, Oxidoreductase, Glycine Soja Protein], Pepha-Timp [INCI: Human Oligopeptide-20], Pepha-Age [INCI: Dunaliella Salina Extract], Colhibin [INCI: Hydrolyzed Rice Protein], Elhibin [INCI: Glycine Soja protein, disodium cocoamphodiacetate], or All-Q™ Plus [INCI: ubiquinone, tocopheryl acetate]; Myoxinol™ [INCI: hydrolyzed Hibiscus esculentus extract], Myoxinol™ LS 9736 [INCI: hydrolyzed Hibiscus esculentus extract, dextrin], Syniorage™ [INCI: acetyl tetrapeptide-11], Dermican™ [INCI: acetyl tetrapeptide-9], DN-AGE® LS [INCI: Cassia alata leaf extract], Hyalufix GL [INCI: Alpinia Galanga leaf extract], Neurobiox [INCI: Achillea Millefolium extract], Deliner [INCI: Zea Mays (corn) seed extract], Lys'lastine, marketed by Laboratoires Serobiologiques / Cognis / BASF. V [INCI: Peucedanum Graveolens (Dill) Extract], Extracellium [INCI: Hydrolyzed Potato Protein], Proteasyl TP LS 8657 [INCI: Pisum Sativum Extract], FlavagrumPEG [INCI: PEG-6 isostearate, hesperetin laurate], Micromerol [INCI: Pyrus Malus fruit extract], Extracellium [INCI: Hydrolyzed potato protein], Marine Filling Spheres [INCI: Pentaerythrityl tetraisostearate, dimethylsilylated silica, sodium chondroitin sulfate, atelocollagen], Triactigen [INCI: Mannitol, cyclodextrin, yeast extract, disodium succinate], Eterniskin [INCI: Grifola Frondosa fruit body extract, maltodextrin], Ascotide [INCI: Ascorbyl succinoyl pentapeptide-12 phosphate], Hyalurosmooth [INCI: Cassia Angustifolia seed polysaccharide], Indinyl CA [INCI: Cassia Angustifolia seed polysaccharide], Arganyl [INCI: Argania Spinosa Leaf Extract], Sphingoceryl Veg [INCI: Plant Ceramides], Vit-A-Like [INCI: Vigna Acontifolia Seed Extract], Peptiskin [INCI: Arginine / Lysine Polypeptide], Prodejine [INCI: Mannitol, Cyclodextrin, Yeast Extract, Disodium Succinate], Aqu'activ [INCI: Behenyl Alcohol, Glyceryl Oleate, Cocamide MIPA, Calcium Citrate], Elestan [INCI: Glycerin, Manilkara Leaf Extract], Hibiscin HP [INCI: Hibiscus Esculentus Seed Extract], Collalift® 18 [INCI: Khaya Senegalensis Skin], Collrepair® DG [INCI: Hexylene Glycol, Niacin], or Litchiderm [INCI: Litchi ChinensisPericarp Extract]; Argireline® [INCI: Acetyl Hexapeptide-8], SNAP-7 [INCI: Acetyl Heptapeptide-4], SNAP-8 [INCI: Acetyl Octapeptide-3], Leuphasyl® [INCI: Pentapeptide-18], Inyline® [INCI: Acetyl Hexapeptide-30], Aldenine® [INCI: Hydrolyzed Wheat Protein, Hydrolyzed Soy Protein, Tripeptide-1], Preventhelia ( Decorinyl® [INCI: Tripeptide-10 Citrulline], Decorinol® [INCI: Tripeptide-9 Citrulline], Trylagen® [INCI: Pseudoalteromonas Ferment Extract, Hydrolyzed Wheat Protein, Hydrolyzed Soy Protein, Tripeptide-10 Citrulline, Tripeptide-1], Eyeseryl® [INCI: Acetyl Tetrapeptide-5], Peptide AC29 [INCI: Acetyl Tripeptide-30 Citrulline], Relistase® [INCI: Acetyl Arginyl Tryptophyl Diphenylglycine], Thermostressine® [INCI: Acetyl Tetrapeptide-22], Lipochroman® [INCI: Dimethyl Methoxychromanol], Chromabright® [INCI: Dimethyl Methoxychromanyl Palmitate], Antarcticine® [INCI: Pseudoalteromonas spp.] Ferment Extract], dGlyage® [INCI: Lysine HCl, Lecithin, Tripeptide-9 Citrulline], Vilastene™ [INCI: Lysine HCl, Lecithin, Tripeptide-10 Citrulline], Hyadisine® [INCI: Pseudoalteromonas Ferment Extract], Hyanify™ [INCI: Saccharide Isomerates], Diffuporine® [INCI: Acetyl Hexapeptide-37], Silusyne® [INCI: Soy (GlycineSoja (Soybean) Oil, Sorbitan Sesquioleate, Isohexadecane, Sodium Hyaluronate, Lauryldimonium Hydroxypropyl Hydrolyzed Soy Protein, Acetyl Hexapeptide-39], Adifyline® [INCI: Acetyl Hexapeptide-38], Delisens™ [INCI: Acetyl Hexapeptide-46], Telangyn™ [INCI: Acetyl Tetrapeptide-40], Reproage™ Peptides [INCI: Acetyl Hexapeptide-8], Cellynkage™ Marine Ingredients [INCI: Saccharide Isomerates], Eyedeline™ Marine Ingredients [INCI: Plankton Extract], Uplevi ty™ [INCI: Acetyl tetrapeptide-2], Seacode™ Marine Ingredients [INCI: Pseudoalteromonas ferment extract], or Serilesine™ Peptide Solution [INCI: Hexapeptide-10], marketed by Lipotec / Lubrizol; Argireline™ Amplified [INCI: Acetyl hexapeptide-8], Dawnergy™ [INCI: Nonapeptide-1], marketed by Lipotrue; Sirtalice™ [INCI: Bacillus ferment], Epitensive™ [INCI: Nicotiana benthamiana hexapeptide-40], marketed by Lipotrue; SH-Oligopeptide-1], Scelleye™ [INCI: Nicotiana benthamiana SH-Oligopeptide-2], Seadermium [INCI: Aqua, Glycerin, Bacillus ferment], Pauseile [INCI: Aqua, Glycerin, Bacillus ferment], or Neoclair pro [INCI: Aqua, Glycerin, Caprylyl glycol, Acetyl tetrapeptide-2]; Collaxyl® IS [INCI: Hexapeptide-9], Laminixyl IS™ [INCI: Heptapeptide], Orsirtine™ GL [INCI: Oryza sativa (Rice) extract], D'Orientine™ IS [INCI: Phoenix dactylifera (Date Palm) Seed Extract], Phytoquintescine™ [INCI: Einkorn (Einkorn) Extract], Quintescine™ IS [INCI: Dipeptide-4], Peptide Vinci 01 [INCI: Pentadecapeptide-1], Peptide Vinci 02™ [INCI: Hexapeptide-3], Aquarize IS™ [INCI: Hydrolyzed Rice Extract], Lanablue [INCI: Algae Extract], Ederline™ [INCI: Pyrus Malus (Apple) Seed Extract], Dynachondrine™ ISR [INCI: Hydrolyzed Soy Protein], Prolixir S20™ [INCI: Dimeric Tripeptide-43], Phytocohesine™ PSP [INCI: Sodium Beta-Sitosteryl Sulfate, Beta-Sitosterol], Perenityl™ IS [INCI: Pyrus Communis (Pear) Seed Extract], Caspaline 14™ [INCI: Hexapeptide-42], Peptide Q10™ [INCI: Pentapeptide-34 Trifluoroacetate], Survixyl IS™ [INCI: Pentapeptide-31], ChroNOgen™ [INCI: Tetrapeptide-26], Elixiance [INCI: SchinusMolle extract], Harmoniance™ [INCI: Nelumbo Nucifera flower extract], Serenityl [INCI: Marsdenia Condurango peel extract], Natriance Wrinkle-less [INCI: Hydrolyzed corn protein], Phytoneomatrix [INCI: Hydrolyzed soy extract], Prolixir ICE [INCI: Hydrolyzed rice protein], PhytoRNx Baobab™ [INCI: Hydrolyzed Adansonia Digitata extract], Natriance Renovate extract [INCI: Hydrolyzed flaxseed extract], Natriance Self-hydrate extract [INCI: Pisum Sativum extract], Actopontine YST [INCI: Hydrolyzed yeast protein], or Telosense™ [proposed INCI: Hydrolyzed soy protein, hydrolyzed yeast protein]; BONT-L-Peptide [INCI: Palmitoyl hexapeptide-19], TIMP marketed by Infinitec Activos. Peptide [INCI: Acetyl Hexapeptide-20], ECM Moduline [INCI: Palmitoyl Tripeptide-28], Renaissance [INCI: Hydrolyzed Wheat Protein, Palmitoyl Decapeptide-21, Decapeptide-22, Oligopeptide-78, Zinc Palmitoyl Nonapeptide-14], or X50 Antiaging [INCI: Lactic / Glycolic Acid Copolymer, Polyvinyl Alcohol, Copper Palmitoyl Heptapeptide-14, Heptapeptide-15 Palmitate]; EquiStat [INCI: Pyrus malus fruit extract, Glycine max. soja seed extract], Juvenesce [INCI: Ethoxydiglycol and Caprylic Triglycerides, Retinol, Ursolic Acid, Phytonadione, Ilomastat], Ursolisome [INCI: Lecithin, Ursolic Acid, Atelocollagen, Xanthan Gum, Sodium Chondroitin Sulfate], Basaline [INCI: Hydrolyzed Malt Extract], Phytokine [INCI: Hydrolyzed Soy Protein]; MibelleAmeliox [INCI: Carnosine, Tocopherol, Silybum marianum fruit extract], or PhytoCellTec Malus Domestica [INCI: Malus domestica fruit cell culture], Lipobelle Soyaglicane [INCI: Soy isoflavones], RoyalEpigen P5 [INCI: Butyrospermum Parkii butter, hydrogenated lecithin, maltodextrin, pentapeptide-48, phenethyl alcohol, ethylhexylglycerin, glycerin, aqua], or DermCom [INCI: Crocus Chrysanthus bulb extract, gum arabic, aqua / water], marketed by Biochemistr; ActiMatrix [INCI: Peptide-based mushroom extract], Peptamide 6 [INCI: Hexapeptide-11], marketed by Active Organics / Arch; and combinations thereof.

[0073] The firming, skin elasticity and / or restructuring agents may be Argassential [INCI: C10-16 Alkyl Glucoside, Dicaprylyl Ether, Glycerin] or Replexium BC [INCI: Dimethyl Isosorbide, Polysorbate 20, Aqua, Acetyl Tetrapeptide-11, Acetyl Tetrapeptide-9] marketed by BASF; Prolevis [INCI: Hydrolyzed Vegetable Protein] or Poretect [INCI: Caprylic / Capric Triglyceride, Sorbitan Trioleate, Apium Graveolens Seed Extract, Linum Usitatissimum Seed Extract] marketed by Sederma / Croda; Actifirm Ultra Advanced Botanical Ingredients [INCI: Centella Asiatica Extract Extract, Rosmarinus Officinalis Leaf Extract, Dipropylene Glycol, Alcohol, Echinacea Root Extract] marketed by Lipotec / Lubrizol. Angustifolia Leaf Extract], or Actifcol Advanced Botanical Ingredients [INCI: Aqua, Glycerin, Sodium Citrate, Lentinus Edodes Extract, Potassium Sorbate, Sodium Benzoate, Phytic Acid] sold by Mibelle; Densorphin™ [INCI: Vitex Agnus Castus Extract, Aqua, Maltodextrin] or PhytoCellTec™ nunatak® [INCI: Isomalt, Aqua, Saponaria Pumila Callus Culture Extract, Lecithin]; and combinations thereof.

[0074] Moisturizers include qua Shuttle [INCI: Sorbitol, Laminaria Digitata extract, Diatomaceous Earth] marketed by Infinitec; Aqua-Osmoline™ [INCI: Ceratonia Siliqua (Carob) seed extract] marketed by Vincience / ISP / Ashland; Hydralphatine™ Asia [INCI: Hydrogenated Starch Hydrolysate, Panthenol, Bambusa Vulgaris Shoot Extract, Nelumbo Nucifera Flower Extract, Nymphaea Alba root extract], or Hydraporine™ [INCI: betaine, hydrogenated lecithin, honey, pectin]; PatcH2O™ [INCI: trehalose, urea, serine, glycerol polyacrylate, algin, sodium hyaluronate, pullulan], marketed by L. Serobiologiques / Cognis / BASF, Aqu'activ™ [INCI: behenyl alcohol, glyceryl oleate, cocamide MIPA], Irwinol® [INCI: octyldodecanol, Irvingia Gabonensis kernel butter, hydrogenated coco-glycerides], Lipodermole® [INCI: Octyldodecanol, Arachidyl propionate, Tocopheryl acetate, Retinyl palmitate, Ethyl linoleate, Ethyl linolenate] or Seanamin® SU [INCI: Sorbitol, algae extract, Chrondrus crispus (carrageenan), Fucus vesiculosus extract, algin]; Snow algae powder [INCI: Coenochloris signiensis extract] marketed by Mibelle; Hyasol BT [INCI: Sodium hyaluronate], Syn-Up™ [INCI: Benzylsulfonyl D-seryl homophenylalanine amidinobenzamide acetate] or Pentavitin® [INCI: Saccharide isomerate] marketed by Pentapharm / DSM;Aqualance™ [INCI: Erythritol, Homarine HCl], Hydraprotectol™ [INCI: Glyceryl Polymethacrylate, Aryl Acid, Yeast Extract (Faex), Glycoproteins], Moist 24™ [INCI: Imperata Cylindrica Root Extract], Optimum™ [INCI: Imperata Cylindrica Root Extract], marketed by Sederma / Croda Hyal™ [INCI: Hydrolyzed Yeast Extract, Cetyl Hydroxyethylcellulose, Polyglucuronic Acid], Osmocide® 4 [INCI: Glycerin, Acrylic Acid / C10-30 Alkyl Acrylate Crosspolymer], or Revidrate™ [INCI: Ethylhexyl Palmitate, Sorbitan Oleate, Sorbitan Laureate, Myristylphosphonic Malate]; Xpertmoist® Molecular Film [INCI: Glycerin, Pseudoalteromonas Ferment Extract, Xanthan Gum, Proline, Alanine, Serine, Ethylhexylglycerin, Caprylyl Glycol], or Actizyme GL Advanced Botanical Ingredients [INCI: Glycerin, Mucor miehei Extract, Aqua, Sodium Citrate, Potassium Sorbate, Sodium Benzoate, Phytic Acid], marketed by Lipotec / Lubrizol; and combinations thereof.

[0075] Anti-photoaging and / or blue light protection agents include Algaktiv Genofix CPD marketed by Greenaltech [INCI: Plankton Extract, Aqua, Lecithin]; Blumilight™ Biofunctional marketed by Ashland [Proposed INCI: Water / Aqua (and) Butylene Glycol (and) Theobroma Cacao (Cocoa) [Seed Extract]; Lys'Sun marketed by BASF [INCI: Hamamelis Virginiana Leaf Extract, Aqua, Pentylene Glycol, Caprylyl Glycol, Xanthan Gum]; Vitachelox marketed by Indena [INCI: Vitis Vinifera Seed Extract, Camellia Sinensis Leaf Extract, Quercus Robur Wood Extract]; Freshine L-VCG [INCI: Ascorbyl Glucoside] marketed by Bio-technology; Lumicease Blue Ingredients [INCI: Glycerin, Aqua, Hydrolyzed Pea Protein, Glucose, Sodium Chloride] marketed by Lipotec / Lubrizol; Lightwaves Defense [JS+M] [INCI: Jasminum Sambac Leaf Cell Extract] marketed by Naolys; Blue Oleoactif [INCI: Glycine Soja Oil, Polyglyceryl-3 Diisostearate, Oryza Sativa Germ Extract, Oryza Sativa Extract] marketed by Oleos-Hallstar; Majestem [INCI: Glycerin, Leontopodium Alpinum Callus Culture Extract, Xanthan Gum] or Senestem [INCI: Glycerin, Plantago sativa L. Extract] marketed by Sederma Lanceolata leaf extract, xanthan gum]; Blueshield [INCI: Glycerin, Capsicum Annuum fruit extract, xanthan gum], marketed by Solabia; and combinations thereof.

[0076] DNA protectants, DNA repair agents, and / or stem cell protectants include GP4G SP [INCI: Aqua, Glycerin, Aretmia extract], Heliostatine [INCI: Aqua, Glycerin, Pisum Sativum extract], Orsirtine [INCI: Aqua, Glycerin, Oryza Sativa extract], Chronogen [INCI: Aqua, Butylene glycol, Tetrapeptide (proposed INCI)], Survixyl IS [INCI: Water, Butylene glycol, Pentapeptide-31], and Chrondricare [INCI: Aqua, Butylene glycol, Pentapeptide-28] marketed by Vincience / ISP / Ashland; Lanacityn® [INCI: Glycerin, Aqua, Alteromonas ferment extract, Chysanthellum extract] marketed by Atrium Innovations / Lucas Meyer Cosmetics; indicum extract] or Melinoil [INCI: Isopropyl palmitate, lecithin, aqua, acetyl hexapeptide-1]; Repair Complex [INCI: Bifida ferment lysate], marketed by CLR; Phycojuvenine [INCI: Laminaria Digitata], marketed by Codif; Unirepair T-43 [INCI: Butylene glycol, acetyl tyrosine, proline, hydrolyzed vegetable protein, adenosine triphosphate], marketed by Induchem; Dragosine [INCI: Carnosine], marketed by Symrise; DN-Age [INCI: Cassia Alata leaf extract], marketed by Laboratories Serobiologiques / Cognis / BASF;Helioguard [INCI: Porphyra Umbilicalis Encapsulated in Liposomes], PhytoCellTec Malus Domestica [INCI: PhytoCellTec Malus Domestica], or PhytoCellTec Argan [INCI: Argania Spinosa Sprout Cell Extract, Isomalt, Lecithin, Sodium Benzoate, Aqua], marketed by Mibelle Biochemistry; Pepha-Protect [INCI: Watermelon Extract], marketed by Pentapharm / DSM; Celligent [INCI: Helianthus Annuus Seed Oil, Ethyl Ferulate, Polyglyceryl-5 Trioleate, Rosmarinus Officinalis Leaf Extract, Aqua, Disodium Uridine Phosphate], or Defensil [INCI: Octyldodecanol, Echium Plantagineum Seed Oil, Cardiospermum Halicacabum Extract, Helianthus Annuus Seed Oil Unsaponifiables]; marketed by Sederma / Croda, Venuceane [INCI: Thermus Thermophilus Ferment, Glycerin], UV-Soft [INCI: Yeast Extract], Renovage [INCI: Caprylic / Capric Triglyceride, Teprenone], Juvinity [INCI: Caprylic / Capric Triglyceride, Geranylgeranylpropanol (suggested)], Phytessence Holyherb [INCI: Butylene Glycol, Eriodictyon Californicum (Holyherb) Flower / Leaf / Stem Extract], or Resistem [INCI: Glycerin, Globularia Cordifolia Ferment]; marketed by Mibelle, Infraguard [INCI: Caesalpinia Spinosa Fruit Pod Extract, Propylene Glycol, Aqua, Helianthus Annuus Bud Extract, Sodium Benzoate, Phenoxyethanol];Heliomoduline [INCI: low molecular weight peptides from cotton] or Stem-C-Guard [hydrolyzed peas], marketed by Silab; and combinations thereof.

[0077] Reactive carbonyl species scavengers, free radical scavengers, and / or anti-glycation agents, detoxifying agents, antioxidants, and / or anti-pollutants include, but are not limited to, carnosine and its derivatives; GHK [INCI: Tripeptide-1] and its salts and / or derivatives, or Quintescine IS [INCI: Dipeptide-4], marketed by Vincience / ISP / Ashland; Preregen [INCI: Glycine Soja Protein, Oxidoreductase], Edelweiss GC [INCI: Leontopodium Alpinum Extract], Lipogard [INCI: Squalane, Ubiquinone], Nectapure [INCI: Buddleja Davidii Extract, Thymus Vulgaris Extract], Alpaflor Nectapure [INCI: Buddleja Davidii Extract, Thymus Vulgaris extract, glycerin, water], or Dismutin-BT [INCI: Highly purified SOD from a natural yeast strain of Saccharomyces cerevisiae]; Preventhelia® [INCI: Diaminopropionoyl tripeptide-33], Aldenine® [INCI: Hydrolyzed wheat protein, hydrolyzed soy protein, tripeptide 1], Lipochroman™ [INCI: Dimethylmethoxychromanol], Thermostressine® [INCI: Acetyl tetrapeptide-22], Pollushield™ Functional Ingredients [INCI: Diisopropyl adipate, lecithin, acrylic acid / acrylamidomethylpropanesulfonic acid copolymer, dimethylmethoxychromanol, xanthan gum], or Bodyfensine® [INCI: Acetyl aminohexanoate dipeptide-3], marketed by Lipotec / Lubrizol;unactyl [INCI: Mannitol, Pisum Sativum Extract, Histidine HCl, Arginine, Cyclodextrin, Dextrin, Yeast Extract, Acetyl Trypsin, Pyridoxine HCl, Khaya Senegalensis Peel Extract, Nicotinamide, Adenine Dinucleotide, Disodium Succinate, Aspartic Acid], Imidinyl [INCI: Tamarindus Indica Seed Polysaccharide], Phystrogene [INCI: Butylene Glycol, Malva Sylvestris (Mallow) Extract, Xanthan Gum], or Purisoft [INCI: Moringa Pterogysperma Seed Extract], marketed by Laboratoires Serobiologiques / Cognis / BASF; AquaCacteen [INCI: Glycerin, Opuntia Ficus Seed Extract], marketed by Mibelle Biochemistry; Indica Stem Extract, Phenoxyethanol, Aqua], Trimoist (KMF) [INCI: Sodium Stearoyl Lactylate, Cetyl Alcohol, Olus Vegetable Oil, Tocopheryl Acetate, Glycerin, Glycine soja Sterol, Sodium Lactate, Sodium Barboxymethyl Beta Glucan, Carnosine, Lactic Acid], MelanoBronze [INCI: Vitex Agnus Castus Extract (Monks Pepper Fruit Extract (Phytoendorphin)), Acetyl Tyrosine], CM-Glucan [INCI: Sodium Carboxymethyl Beta Glucan, Phenoxyethanol, SunActin [INCI: Helianthus Annuus (Sunflower) Sprout Extract], Tocopherol, Glycerin, Lecithin, Phenoxyethanol, Aqua], GSP-T skin [INCI: Glycerin, Alcohol, Aqua, PEG-40 Hydrogenated Castor Oil, Vitis Vinifera (Grape) Seed Extract], or Detoxophane [INCI: Lepidium Sativum Bud Extract, Lecithin, Phenoxyethanol, Glycerin, Water];Bacocalmine [INCI: PEG-8, Bacopa Monniera extract, water (aqua), hydroxyethylcellulose], Kombuchka [INCI: Saccharomyces / Xylinum black tea ferment, glycerin, hydroxyethylcellulose], Citystem [INCI: Glycerin, Marrubium Vulgare extract], or Prodizia [INCI: Albizia Julibrissin extract, glycerin], marketed by Sederma / Croda; Extramel C [INCI: Hydroxypropyltrimonium Maltodextrin Crosspolymer, Cucumis Melo (Melon)] fruit extract, marketed by Seppic; Defensine [INCI: Triticum Vulgare Germ Extract], Apolluskin® [INCI: Taraxacum officinale (Dandelion) Extract], Detoxyl® [INCI: Water, Butylene Glycol, Butyrospermum Extract], marketed by Silab parkii (Shea Butter) Seed Cake Extract], or Antiglyskin [INCI: Aqua, Helianthus Annuus Seed Extract; and combinations thereof.

[0078] The compositions of the present invention may also contain other active ingredients, such as vitamins, minerals, proteins, peptides, fatty acids, antioxidants, anti-inflammatory agents, darkening agents, and / or mixtures thereof. In particular, the cosmetic compositions may contain other active ingredients for promoting hair growth and / or preventing hair loss. Non-limiting examples suitable in the context of the present invention are Growth Oleoactif® marketed by Hallstar [INCI: Helianthus Annuus seed oil (and) Polyglyceryl-3 diisostearate (and) Carthamus Tinctorius (safflower) (and) Hibiscus Sabdariffa flower extract]; Widelash™ marketed by Sederma [INCI: Glycerin, Water (Aqua), Panthenol, Biotinoyl Tripeptide-1]; SymPeptide® Xlash marketed by Symrise [INCI: Glycerin (and) Aqua (and) Myristoyl Pentapeptide-17], SymLash® 1631 [INCI: Pentylene glycol (and) Isochrysis Galbana (and) Galbana Extract]; Anargy™, available from Lipotrue [INCI: Aqua (and) Butylene Glycol (and) Oligopeptide-2 (and) Nicotiana Benthamiana Hexapeptide-40 sh-polypeptide-9 (and) Nicotiana Benthamiana Hexapeptide-40 sh-polypeptide-86]; Capixyl™, available from Lucas Meyer [INCI: Butylene Glycol (and) Aqua (and) Dextran (and) Acetyl Tetrapeptide-3 (and) Trifolium Pratense (Clover) Flower Extract]; Nano lashes, available from Nanovetores [INCI: Aqua, Simmondsia Chinensis Seed Extract, Polysorbate 20, Hydroxypropyl Guar, Sodium Benzoate, Potassium Sorbate];Marketed by Spec-chem are Specped SC-MH16 [INCI: Aqua, Glycerin, Myristoyl Hexapeptide-16, Caprylyl Glycol, Ethylhexylglycerin], Myristoyl Pentapeptide-17 [INCI: Myristoyl Pentapeptide-17], SpecPed® BT1 [INCI: Biotinoyl Tripeptide-1], SpecPed® LashLD [INCI: Biotinoyl Tripeptide-1, Glycerin, Water, Caprylyl Glycol and Ethylhexylglycerin, Panthenol, Trehalose], SpecPed® MP17P [INCI: Myristoyl Pentapeptide-17]; Procapil™ marketed by Sederma [INCI: Butylene Glycol (and) Aqua (and) PPG-26-Buteth-26 (and) PEG-40 Hydrogenated Castor Oil (and) Apigenin (and) Oleanolic Acid (and) [Biotinoyl Tripeptide-1]; AnaGain™ marketed by Mibelle [INCI: Pea (Pisum Sativum (Pea) Sprout Extract (and) Isomalt (and) Aqua], PhytoCellTec™ Malus Domestica Hair [INCI: Malus Domestica Fruit Cell Culture Extract (and) Xanthan Gum (and) Glycerin (and) Lecithin (and) Phenoxyethanol (and) Aqua / Water], Santenergy™ [INCI: Bioflavonoids (and) Pentylene Glycol (and) Alcohol (and) Aqua], RootBioTec™ HW [INCI: Ocimum Basilicum Hair Root Culture Extract (and) Alcohol (and) Aqua / Water], RootBioTec™ HO [INCI: Basil Hair Root Culture Extract (and) Helianthus Annuus (Sunflower) Seed Oil (and) Cocos Nucifera (Coconut) Oil; CAPILIA LONGA marketed by Vytrus Biotech PPF [INCI: Turmeric (Curcuma longa) callus culture conditioned medium (and)] water];marketed by Solabia, [INCI: Glycerin (and) Aqua (and) Nasturtium Officinale Extract (and) Tropaeolum Majus Extract]; marketed by Peptron, Alotide™, [INCI: Copper Ascorbyl Phosphate Succinyl Tripeptide-34]; marketed by Ichimaru Pharcos, BURGEON-UP, [INCI: Aqua (and) Alcohol (and) Nasturtium Leaf / Stem Extract]; marketed by Exysmol, Capalgin®, [INCI: Chondrus Crispus Extract]; marketed by BASF, Dermosaccharides® GY, [INCI: Aqua (and) Glycerin (and) Glycogen (and) Phenoxyethanol (and) Methylparaben], Trichogen™ VEG LS 9922, [INCI: Water (and) Panax Ginseng (and) Ginseng (Ginseng) Root Extract (and) Arginine (and) Acetyl Tyrosine (and) Burdock (Arctium Majus) Root Extract (and) Hydrolyzed Soy Protein (and) Polyquaternium-11 (and) PEG-12 Dimethicone (and) Calcium Pantothenate (and) Zinc Gluconate (and) Niacinamide (and) Ornithine HCl (and) Citrulline (and) Glucosamine HCl (and) Biotin], marketed by CLR; Follicusan™ DP [INCI: Aqua (and) Alcohol Denat. (and) Panthenyl Ethyl Ether (and) Inositol (and) Milk Protein (and) Lactose (and) Acetyl Cysteine ​​(and) Acetyl Methionine (and) Sodium Citrate (and) Citric Acid], marketed by Greentech; Hairline® [INCI: Propanediol (and) Water (Aqua) (and) Lindera Strychnifolia) root extract];Kerascalp™ [INCI: Propanediol (and) Glycerin (and) Phyllanthus emblica Fruit Extract], marketed by Provital, Baicapil™ [INCI: Propanediol (and) Water (and) Arginine (and) Lactic Acid (and) Glycine Soja (Soybean) Germ Extract (and) Triticum Vulgare (Wheat) Germ Extract (and) Scutellaria Baicalensis Root Extract (and) Sodium Benzoate (and) Gluconolactone (and) Calcium Gluconate], marketed by Seppic; BIOENERGIZER™ P BG PF [INCI: Aqua / Water, Butylene Glycol, Panthenol, Propylene Glycol, Pelvetia Canaliculata Extract, Laminaria Digitata Extract, Protectagen™ (INCI: Aqua (and) Glycerin (and) Hydrolyzed Rice Protein) marketed by Ashland; Redensyl® (INCI: Glycerin (and) Aqua (and) Sodium Metabisulfite (and) Larix Europaea Extract (and) Glycine (and) Zinc Chloride (and) Camellia Sinensis Leaf Extract) marketed by Givaudan; GANOTHER® (INCI: Glycerin (and) Aqua (and) Ganoderma Lucidum Mycelium Ferment Filtrate) marketed by B&G; Hairgenyl® (INCI: Saccharomyces Cerevisiae Extract), Anageline® (INCI: Hydrolyzed Lupin Protein) marketed by Silab;Hairdian AP marketed by Shanghai GREAF Biotech [INCI: Propanediol (and) Thuja Orientalis Extract (and) Zingiber Officinale (Ginger) Root Extract (and) Trifolium Pratense (Clover) Leaf Extract (and) Artemisia Argyi Leaf Extract], Naturein® Wheat Peptides (INCI: Water (Aqua) (and) Hydrolyzed Wheat Protein) from Givaudan, or KeraMatch® V from Croda Personal care (INCI: Water (and) Hydrolyzed Pea Protein (and) Hydrolyzed Vegetable Protein);

[0079] Purpose The present invention is based on the finding that the protein hydrolysate of the present invention is effective in reducing and / or preventing the harmful effects of exposomes on skin and / or hair. The harmful effects of exposomes include the harmful effects caused by air pollution, sunlight, and / or greenhouse gases. The protein hydrolysate has also been found to be effective in reducing the harmful effects of chemical treatments or heat stress on hair. In addition, the protein hydrolysate has been found to be effective in maintaining and / or improving the physical barrier function of the skin. The physical barrier function of the skin is also referred to herein as the skin physical barrier, the skin permeability barrier, or simply the skin barrier. This barrier is provided by tight junctions in the stratum corneum and the epidermis.

[0080] In one aspect, the present invention relates to a protein hydrolysate as described herein for the treatment and / or care of the skin and / or hair, or (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and The present invention provides the use of stereoisomers and cosmetically acceptable salts thereof, or a combination of said protein hydrolysate or said compound with a filtrate obtained by fermentation of the roots of Raphanus sativus by the microorganism Leuconostoc, in particular for the treatment and / or care of the skin and / or hair, both cosmetic and non-therapeutic.

[0081] The use of the protein hydrolysates, compounds, or combinations described herein can be for reducing and / or preventing the harmful effects of exposomes on hair, reducing and / or preventing the harmful effects of air pollution on hair, reducing and / or preventing the harmful effects of sunlight on hair, reducing and / or preventing the harmful effects of chemical stress on hair, reducing and / or preventing the harmful effects of heat stress on hair, and / or reducing or preventing the harmful effects on hair caused by lipid peroxidation in hair and / or protein carbonylation in hair. The use of the protein hydrolysates, compounds, or combinations described herein can be for reducing and / or preventing the harmful effects of air pollution and sunlight on hair. The harmful effects on hair can be manifested in the form of damaged hair. Thus, the protein hydrolysates can be used to reduce hair damage. The harmful effects on hair can be manifested in the form of hair with more split ends, reduced fiber alignment, and / or increased frizz. Thus, the hair is more difficult to manage. Thus, protein hydrolysates may be used to increase hair manageability, in particular by reducing split ends and / or increasing fiber alignment and / or reducing frizz and / or reducing volume.

[0082] Thus, there is provided the use of a protein hydrolysate, compound or combination as described herein for the non-therapeutic cosmetic treatment and / or care of hair, wherein the treatment and / or care of hair is the reduction or prevention of adverse effects of exposomes on hair.

[0083] There is provided the use of a protein hydrolysate, compound or combination as described herein for the non-therapeutic cosmetic treatment and / or care of hair, wherein the treatment and / or care of hair is the reduction or prevention of the harmful effects of air pollution on hair, in particular where the air pollution contains particulate matter.

[0084] Air pollution includes, for example, particulate matter. Particulate matter is airborne pollutants that include solid particles. Particulate matter can include dust, dirt, soot, or smoke. Particulate matter can come from both anthropogenic and natural sources, including automobiles (e.g., exhaust emissions from automobile combustion engines), power plants, factories, fires, wood-burning stoves, dust from open land or construction, road dust (e.g., brake, tire, and road wear particles), pollen (e.g., cedar pollen), and tobacco smoke (e.g., cigarette smoke). Particulate matter can have a size of 10 microns or less (PM 10 and "dust"). Particulate matter can come from automobile combustion engines (e.g., diesel particulates, such as those encountered in diesel particulate matter NIST 1650B). Particular matter can include pollen (e.g., cedar pollen).

[0085] There is provided the use of a protein hydrolysate, compound or combination as described herein for the non-therapeutic cosmetic treatment and / or care of hair, wherein the treatment and / or care of hair is the reduction or prevention of the harmful effects of air pollution containing particulate matter on hair, wherein the particulate matter is from road dust, from diesel or is pollen.

[0086] There is provided the use of the protein hydrolysates, compounds or combinations described herein for the non-therapeutic cosmetic treatment and / or care of hair, wherein the treatment and / or care of hair is the reduction or prevention of the harmful effects of sunlight on the hair. Sunlight may include UV-A radiation.

[0087] There is provided the use of a protein hydrolysate, compound or combination as described herein for the non-therapeutic cosmetic treatment and / or care of hair, wherein the treatment and / or care of hair is the reduction or prevention of the adverse effects of bleaching or dyeing on the hair.

[0088] There is provided the use of a protein hydrolysate, compound or combination as described herein for the non-therapeutic cosmetic treatment and / or care of hair, wherein the treatment and / or care of hair is the reduction or prevention of the adverse effects of heat stress on hair.

[0089] There is provided the use of a protein hydrolysate, compound or combination as described herein for the non-therapeutic cosmetic treatment and / or care of hair, wherein the treatment and / or care of hair is the reduction or prevention of adverse effects on hair caused by lipid peroxidation in hair and / or protein carbonylation in hair.

[0090] There is provided the use of a protein hydrolysate, compound or combination as described herein for the non-therapeutic cosmetic treatment and / or care of hair, wherein the treatment and / or care of hair is increasing the manageability of the hair, and / or increasing the fiber alignment, and / or reducing frizz, and / or reducing volume.

[0091] The use of the protein hydrolysates, compounds, or combinations described herein can be for reducing and / or preventing the harmful effects of exposomes on the skin, reducing and / or preventing the harmful effects of air pollution on the skin, reducing and / or preventing the harmful effects of sunlight on the skin, reducing and / or preventing the harmful effects caused by greenhouse gases, and / or maintaining and / or improving the physical barrier function of the skin. The use of the protein hydrolysates, compounds, or combinations described herein can be for reducing and / or preventing the harmful effects of air pollution and sunlight on the skin. The harmful effects on the skin can be manifested in the form of a reduction in barrier function, a dull appearance of the skin, loss of skin brightness, luminosity, and / or luster, loss of homogeneity in skin texture, an increase in skin roughness, the appearance of dark spots, and / or erythema (skin redness).

[0092] Thus, there is provided the use of a protein hydrolysate, compound or combination as described herein for non-therapeutic cosmetic treatment and / or care of the skin, wherein the treatment and / or care of the skin is the reduction or prevention of adverse effects of exposomes on the skin.

[0093] There is provided the use of a protein hydrolysate, compound or combination as described herein for the non-therapeutic cosmetic treatment and / or care of the skin, wherein the treatment and / or care of the skin is the reduction or prevention of the harmful effects of air pollution containing particulate matter on the skin, wherein the particulate matter is from road dust, from diesel or is pollen.

[0094] There is provided the use of the protein hydrolysates, compounds or combinations described herein for the non-therapeutic cosmetic treatment and / or care of the skin, wherein the treatment and / or care of the skin is the reduction or prevention of the harmful effects of sunlight on the skin. Sunlight may include UV-A radiation.

[0095] The adverse effects may manifest themselves in the form of a loss in the barrier function of the skin.Therefore, there is provided the use of the protein hydrolysates, compounds or combinations described herein for non-therapeutic cosmetic treatment and / or care of the skin, wherein the treatment and / or care of the skin is the maintenance and / or improvement of the physical barrier function of the skin.

[0096] The adverse effects may be manifested in the form of skin having a dull appearance, and / or skin having a less luminous appearance, and / or skin having a less luminous appearance, and / or skin having a less lustrous appearance.There is therefore provided the use of the protein hydrolysates, compounds or combinations described herein for non-therapeutic cosmetic treatment and / or care of the skin, wherein the treatment and / or care of the skin is to increase skin luminosity and / or to increase skin lustre and / or to increase skin luminosity.

[0097] Adverse effects may also result in the appearance of dark spots.Therefore, there is provided the use of the protein hydrolysate, compound or combination described herein for non-therapeutic cosmetic treatment and / or care of skin, wherein the treatment and / or care of skin is to reduce dark spots.In particular, the treatment and / or care can include reducing melanin in skin.

[0098] The adverse effects may manifest in the form of a reduction in skin tone and / or skin texture uniformity and / or an increase in skin roughness.Therefore, there is provided the use of the protein hydrolysates, compounds or combinations described herein for non-therapeutic cosmetic treatment and / or care of the skin, wherein the treatment and / or care of the skin is to increase skin tone and / or to increase skin texture uniformity and / or to reduce skin roughness.

[0099] The adverse effects may be manifested in the form of erythema or skin redness.Therefore, there is provided the use of the protein hydrolysates, compounds or combinations described herein for non-therapeutic cosmetic treatment and / or care of the skin, wherein the treatment and / or care of the skin is to reduce skin redness.

[0100] In one aspect, the present invention relates to a method of treatment and / or care of the skin and / or hair, comprising administering to a subject an azuki bean protein hydrolysate according to the first aspect of the invention, or (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), (iv)Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn - (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and The present invention relates to a method comprising administering a stereoisomer and a cosmetically acceptable salt thereof, or a combination of the compound and a filtrate obtained by fermentation of Raphanus sativus roots with the microorganism Leuconostoc. The treatment and / or care can be a non-therapeutic cosmetic treatment and / or care. The protein hydrolysate, compound or combination described above can be present in a cosmetic composition, for example a cosmetic composition described herein. The method can be for the treatment and / or care of the skin and / or hair described above with respect to the application (use) of the protein hydrolysate, compound and / or combination of the present invention.

[0101] The treatment and / or care is the reduction or prevention of the adverse effects of exposomes on the skin and / or hair, the reduction or prevention of the adverse effects of air pollution on the skin and / or hair, the reduction or prevention of the adverse effects of sunlight on the skin and / or hair, the reduction or prevention of the adverse effects of chemical treatments of hair, the reduction or prevention of the adverse effects of heat stress on hair, and / or the maintenance and / or improvement of the physical barrier function of the skin.

[0102] In the application of the protein hydrolysates, compounds and combinations of the present invention, the treatment and / or care can be preventive in that it functions to protect the skin and / or hair from harmful effects.In this case, the protein hydrolysates, compounds and combinations of the present invention are applied to the skin and / or hair before exposure to a stressor.The treatment and / or care can be reparative or restorative, i.e., the relief has a reparative or restorative function (e.g., with respect to aesthetic properties).In this case, the protein hydrolysates, compounds and combinations of the present invention can be applied to the skin and / or hair during or after exposure to a stressor.

[0103] For the above methods of the present invention, topical or transdermal application may be performed by iontophoresis, sonophoresis, electroporation, mechanical pressure, osmotic gradients, occlusion therapy, microinjection, needle-free injection by pressure, micro-electrical patches, face masks, or any combination thereof.

[0104] For the above-described methods of the present invention, the frequency of application or administration can vary widely depending on the needs of each subject, but the recommended application is once a month to ten times a day, preferably once a week to four times a day, more preferably three times a week to twice a day, and even more preferably once a day.

[0105] The present invention also provides the use of the protein hydrolysate of the present invention in the therapeutic treatment of skin.Accordingly, the present invention provides the protein hydrolysate disclosed herein for use as a medicament.In particular, the present invention provides the protein hydrolysate disclosed herein for use in the prevention or treatment of diseases or disorders associated with exposome, associated with exposure of skin to air pollution, sunlight, and / or greenhouse gases, and / or associated with impaired physical barrier function of the skin.Furthermore, the present invention provides the use of the compound of formula (I), its stereoisomer, and / or its pharma- ceutically acceptable salt for the manufacture of a medicament for the treatment or prevention of diseases or disorders.

[0106] The present invention is further defined by the following non-limiting examples. EXAMPLES

[0107] Example 1 Preparation of ground adzuki beans The first step was to process the raw beans into a fine powder with a brown granular appearance. The aim of this step is to reduce the particle size as much as possible to facilitate protein solubilization. The adzuki beans were ground into a fine flour and the larger fragments were removed by sieving.

[0108] Example 2 enzymatic hydrolysis 2.1 Pretreatment of fine powder Pretreatment of ground azuki beans (also referred to herein as "flour") increases the content of water-soluble proteins in the hydrolysis medium prior to enzyme addition. The flour was suspended in deionized water at a concentration of 21.8 g flour / Kg water (w / w) and transferred into a suitable reaction vessel equipped with pH and temperature probes providing real-time monitoring. The initial pH was raised from 6.5-6.8 to 7.0 by addition of a basic solution (i.e., 1N NaOH) and, when required, more solvent was added to reach the desired concentration. Conditions were maintained at room temperature (25±2°C) for 1 hour. The resulting suspension, referred to as "protein suspension", is a suspension of ground azuki beans in water, which contains solubilized proteins from the ground azuki beans.

[0109] 2.2. Enzymatic hydrolysis The enzyme was added to the protein suspension and the mixture was stirred for 1 hour at room temperature (25±2° C.). The stirring conditions were adjusted to ensure homogeneous and vigorous stirring while keeping foam formation at a minimum level. Alcalase was then added to the protein suspension. Alcalase is a subtilisin type enzyme. The preferred hydrolysis conditions are defined by the use of the subtilisin type Alcalase AF 2.4L (CAS number 9014-01-1, EC 3.4.21.62, Novozymes®) enzyme.

[0110] The proper progress of the hydrolysis was monitored by the change in pH. A decrease in pH is associated with the release of peptide fragments and the subsequent accumulation of acidic carboxylic acids. Typically, the crude hydrolysate undergoes a slight acidification during the process (a decrease in pH units of 0.2-0.3).

[0111] 2.3 Enzyme inactivation After enzymatic hydrolysis, the enzymes were inactivated by increasing the temperature to at least 70° C. and maintaining the temperature for at least 20 minutes. The resulting hydrolysate is hereinafter referred to as “crude hydrolysate”.

[0112] Example 3 purification With the aim of enriching the mixture in the desired active components (ie, peptides), further clarification and dialysis steps can be carried out as described in more detail below.

[0113] Centrifugation. The hydrolyzed azuki bean suspension ("crude hydrolysate") is optimally centrifuged in continuous flow (i.e. using a Westfalia CSA-1 centrifuge or similar equipment) at 10,000 x G with an optimal flow rate of 50 l / h. The supernatant was collected and the pellet was discarded as waste material. Supernatant turbidity was controlled throughout the run taking representative samples and maintained below 0.2 AU / g protein at 600 nm (with respect to appropriate blanks in these conditions).

[0114] 3.2. Front filtration. The supernatant obtained from step 3.1. was passed through filters of various sizes under pump-generated pressure. First, a 1.2 μm pore filter was used to discard the bulk of the larger particles that were not eliminated during centrifugation. This first filtration step is optional, and the purpose is to reduce the number of small pore size filters required. The resulting filtrate was then subsequently filtered through 0.45 μm and 0.22 μm pore size filters.

[0115] 3.3.Ultrafiltration. The filtrate obtained in Example 3.2. was filtered through an ultrafiltration membrane (i.e. Millipore reference P2PLBCC05, 0.5 ml 2 / cassette) to remove fragments with a molecular weight above 3 kDa. The resulting filtrate is also referred to as the "clarified hydrolysate" or "permeate."

[0116] Example 4 Purified hydrolysate characterization Twelve different batches of purified hydrolysates obtained as described in Examples 1-3 were analyzed to determine dry weight, protein content, and antioxidant activity. The dry weight was determined by the weight difference after drying at 110°C. Protein content was determined using a BCA Assay kit. Antioxidant activity was determined by the 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) assay. Briefly, 50 μL of 100 μM DPPH in EtOH was mixed with 50 μL of purified hydrolysate, incubated in the dark at 37° C. for 20 min, and absorbance was measured at 517 nm. The percentage of inhibition is calculated from the following formula:

[0117]

number

[0118] The dry weight of the different batches averaged 2-5 mg dry weight / purified hydrolysate. The protein content ranged from 19 to 35% (w / w). DPPH activity ranged from 74 to 80%. The polyphenol content ranged from 0.15 to 0.30 mg / mL.

[0119] Example 5 In vitro repair effect on follicular papilla cells from the scalp Human follicle dermal papilla cells (HFDPCs) derived from the scalp were cultured in follicular papilla cell growth medium at 37°C and 5% CO2. These cells were treated with 0.1 μg / cm 2The cells were stressed with Particle Matter (PM) (from European Reference Material; ref. CZ100) at 37 °C, followed by UV-A irradiation (LED source, emission peak at 365 nm, 3 J / cm2). The cells were treated with the stressor for 13 min. Immediately after stress exposure, the cells were treated with the purified hydrolysate of Example 3 at 1% (v / v) in culture medium. The antioxidant N-acetyl-cysteine ​​(NAC) was used as a positive control. After 2 h, the intracellular levels of protein carbonylation were measured.

[0120] Protein carbonylation analysis: Carbonylated proteins in hair shafts or HFDPCs were labeled in situ using a fluorescent probe functionalized to specifically bind to carbonyl groups in proteins [Baraibar MA et al., "Proteomic quantification and identification of carbonylated proteins upon oxidative stress and during cellular aging" J Proteomics (2013) 30;92:63-70]. Specific fluorescent signals (relative fluorescent units) per experimental group (n=6) were collected and analyzed with ImageJ software. Relative values ​​(%) of carbonylation levels per experimental group were obtained with respect to the basal control. The repair values ​​(%) were obtained for the experimental groups according to the following formula: Repair value = [(carbonylation stress-carbonylation group X) / (carbonylation stress-carbonylation control)] x 100

[0121] The control group was considered at maximum efficiency (100%) and the stress group at minimum efficiency (0%).

[0122] [Table 1]

[0123] The purified hydrolysate significantly repaired and protected against the harmful effects of particulate matter and UV-A radiation.

[0124] Example 6 In vitro protection assessment in follicular papilla cells from scalp Scalp-derived human follicular hair papilla cells (HFDPCs) were cultured in HFDPC growth medium at 37° C. and 5% CO2. The cells were treated (before stress) with the purified hydrolysate according to Example 3 at 1% (v / v) in culture medium for 24 hours at 37° C. and 5% CO2. The next day, the cells were treated with particulate matter (from European Reference Material; reference number CZ100) at 0.1 μg / cm2 in 1x PBS buffer (phosphate buffered saline, pH 7.4), and Japanese cedar pollen (reference number Japanese cedar pollen-Cj * LG-5280 Euromedex; 5 ng / cm2) followed by stress with UV-A irradiation (LED source, emission peak at 365 nm, 3 J / cm2). Stress treatment was performed for 13 min. After treatment, cells were fixed and carbonyl proteins labeled in situ. Detection of protein oxidation (carbonylation) was performed in fixed cells by epifluorescence microscopy.

[0125] Immediately after treatment and stress exposure, cells (HFDPC) were fixed on the plate with an ethanol / acetic acid solution. Carbonylated proteins were labeled in situ using a fluorescent probe (Ex=647nm / Em=650nm) functionalized to specifically bind to carbonyl groups. Fluorescent images were collected with an epifluorescence microscope (ThermoFisher, Evos M5000) and analyzed with ImageJ software (Schneider, 2012). Image collection for different conditions was achieved using identical acquisition conditions (40x objective).

[0126] Raw images were collected in the range of specific signal intensities from 0 to 65535 and analyzed with ImageJ software. The intensity of carbonylation was determined by the integration of the specific fluorescent signal normalized by the evaluated area.

[0127] Efficacy values ​​(%) were obtained for the experimental groups according to the following formula: % Effectiveness of Sample=[(Oxidation Level Stress-Oxidation Level in Tested Sample) / (Oxidation Level Stress-Oxidation Level Control)]×100

[0128] As a reference, the control group is considered as the maximum efficiency (at 100%) and the stress group as the minimum efficiency (at 0%).

[0129] [Table 2]

[0130] The purified hydrolysate protected against the deleterious effects (carbonylation) caused by the stressor.

[0131] Example 7 Ex vivo protection evaluation in Caucasian human hair shafts (ex vivo) Caucasian human hair shafts (female donor; light brown color) were treated with the hydrolysate of Example 3 (active ingredient solubilized in ultrapure water, leave-on (no washing step)) at 1% (v / v) for 30 minutes at room temperature under gentle movement. After this time, the hair shafts were dried at room temperature and stressed with particulate matter (from European Reference Material; reference number CZ100) at 150 μg / cm2, followed by UV-A irradiation (LED source, emission peak at 365 nm, 84 J / cm2) for 6 hours.

[0132] After stress exposure, protein carbonylation was determined as described in Example 5.

[0133] [Table 3]

[0134] The results confirm the protective effect of the purified hydrolysate.

[0135] Example 8 Ex vivo protection evaluation in Asian hair shafts Asian hair shafts were treated with the purified hydrolysate according to Example 3 at 1% (v / v) (active ingredient solubilized in ultrapure water, leave-on (no washing step)) for 30 minutes at room temperature with gentle movement. After this time, the hair shafts were dried at room temperature by natural evaporation and cedar pollen (reference number cedar pollen-Cj) was applied to the hair shafts. * LG-5280 Euromedex; 500ng / cm2), followed by particulate matter application (150μg / cm2, applied as a powder; from European Reference Material; ref. ERM-CZ100) and UV-A irradiation (LED source, emission peak at 365nm, 84J / cm2). 2 ;6 hours of light exposure) were used for stress.

[0136] Protein carbonylation was measured after stress exposure. Hair shafts from each experimental condition were sampled, cryopreserved in Cryomatrix (OCT), flash frozen in liquid nitrogen, and kept at -80°C until OxiProteome View analysis. Hair shaft cross sections (perpendicular to the hair axis, sagittal view) with a thickness of 3 μm were obtained using a cryostat (Leica) and analyzed. For lateral (cuticle) views, carbonylated proteins were labeled in situ in the entire hair shaft. Carbonylated proteins were labeled in situ using a fluorescent probe (Ex=647nm / Em=650nm) functionalized to specifically bind to carbonyl groups. Fluorescent images were collected with an epifluorescence microscope (ThermoFisher, Evos M5000) and analyzed with ImageJ software (Schneider, 2012). Image collection for the different conditions was achieved using identical acquisition conditions (40x objective). Raw images were collected with a range of specific signal intensities from 0 to 65535 and analyzed with ImageJ software. The intensity of carbonylation was obtained by integration of the specific fluorescent signal normalized by the evaluated area. For hair shaft cross-sections, in each image, quantification of the carbonylation level was obtained independently for the different compartments (cuticle or cortex) and for the whole hair. Three images per condition were used to quantify the carbonylation level. Mean values ​​and standard deviations were obtained. Data processing and statistical analysis were achieved by using graphPad Prism (La Jolla, California, USA).

[0137] Efficacy values ​​(%) were obtained for the experimental groups according to the following formula: Efficacy %(test group x)=[oxidation level (stress)-oxidation level (group x)] / [oxidation level (stress)-oxidation level (control)]×100 As a reference, the control group was considered as the maximum efficiency (at 100%) and the stress group was considered as the minimum efficiency (at 0%).

[0138] [Table 4]

[0139] Example 9 Ex vivo protection evaluation in African human hair shafts African human hair shafts were treated with the adzuki bean hydrolysate of Example 3 at 1% (v / v) (active ingredient solubilized in ultrapure water, leave-on (no washing step)) for 30 minutes at room temperature under gentle movement. After this time, the hair shafts were dried at room temperature and stressed with particulate matter application (150 μg / cm2, applied as a powder; from European Reference Material; ref. ERM-CZ100) and UV-A irradiation (LED source, emission peak at 365 nm, 84 J / cm2). The stress treatment was carried out for 6 hours.

[0140] After stress exposure, protein carbonylation was measured as determined in Example 8.

[0141] [Table 5]

[0142] Example 10 Ex vivo protection evaluation in Latino hair shafts Latin American curly hair shafts were treated with a commercial dye product (chocolate color) having the adzuki bean purified hydrolysate of Example 3 at 1% (v / v) (the active ingredient solubilized in the dye product) for 30 minutes.

[0143] In situ detection of fluorescein diffusion was performed by epifluorescence microscopy or hair cross sections. Hair shafts from each experimental condition were immersed in a fluorophore solution (fluorescein; Ex=498nm / Em=517nm). Excess fluorophore was then eliminated by an extensive washing step with ultrapure water, and hair shafts were sampled, cryopreserved in Cryomatrix (OCT), flash frozen in liquid nitrogen, and kept at -80°C until analysis. Hair shaft cross sections (perpendicular to the hair axis, sagittal view) with a thickness of 3 μm were obtained using a cryostat (Leica) and analyzed. Fluorescence images were collected with an epifluorescence microscope (ThermoFisher, Evos M5000) and analyzed with ImageJ software (Schneider, 2012). Image collection for the different conditions was achieved using identical acquisition conditions (40x objective).

[0144] Raw images were collected with a range of specific signal intensities from 0 to 65535 and analyzed with ImageJ software. The maximum intensity of the diffused fluorophore was obtained and quantified by the integration of the specific fluorescent signal within the transmission axis (from the cuticle to the cortex).

[0145] Three images per condition were used to quantify the fluorescence intensity levels (RFU). Mean values ​​and standard deviations were obtained. Data processing and statistical analysis were achieved by using graphPad Prism (La Jolla, California, USA) and are included in the attached file (Appendix 1-OxiProteomeView-D06CB.xlsx).

[0146] Efficacy values ​​(%) were obtained for the experimental groups according to the following formula and reported in the attached excel file. Efficacy%(Test group X)=[Fluorescence intensity(Stress)-Fluorescence intensity(Test group)] / [Fluorescence intensity(Stress)-Fluorescence intensity(Control)]×100

[0147] As a reference, a control group is considered at maximum efficiency (100%) and a stress group at minimum efficiency (0%).

[0148] [Table 6]

[0149] Example 11 Ex vivo repair efficacy in African human hair shafts African human hair shafts were stressed by heating for 30 minutes at 95° C. After stress exposure, the hair shafts were treated with adzuki bean purified hydrolysate (active ingredient solubilized in ultrapure water) at 1% (v / v) for 30 minutes. In situ detection of fluorescein diffusion was performed by epifluorescence microscopy or hair cross sections.

[0150] Immediately after treatment, hair shafts from each experimental condition were immersed in a fluorophore solution (fluorescein; Ex=498nm / Em=517nm). Excess fluorophore was then eliminated by an extensive washing step with ultrapure water, and hair shafts were sampled, cryopreserved in Cryomatrix (OCT), flash frozen in liquid nitrogen, and kept at -80°C until analysis. Hair shaft cross sections (perpendicular to the hair axis, sagittal view) with a thickness of 3 μm were obtained using a cryostat (Leica) and analyzed. Fluorescent images were collected with an epifluorescence microscope (ThermoFisher, Evos M5000) and analyzed with ImageJ software (Schneider, 2012). Image collection for the different conditions was achieved using identical acquisition conditions (40x objective).

[0151] Raw images were collected with a range of specific signal intensities from 0 to 65535 and analyzed with ImageJ software. The maximum intensity of the diffused fluorophore was obtained and quantified by integration of the specific fluorescent signal in the transmission axis (from cuticle to cortex). Three images per condition were used to quantify the fluorescence intensity level (RFU). Mean values ​​and standard deviations were obtained. Data processing and statistical analysis were achieved by using graphPad Prism (La Jolla, California, USA).

[0152] Efficacy values ​​(%) were obtained for the experimental groups according to the following formula: Efficacy%(Test group X)=[Fluorescence intensity(Stress)-Fluorescence intensity(Test group)] / [Fluorescence intensity(Stress)-Fluorescence intensity(Control)]×100

[0153] The control group is considered at maximum efficiency (100%) and the stress group at minimum efficiency (0%).

[0154] [Table 7]

[0155] Example 12 Protection and defence against lipid peroxidation in hair samples An aqueous solution of the purified hydrolysate according to Example 3 was prepared at 2% (v / v in water) ("active ingredient").

[0156] For protection test: 300mg of hair was weighed into a Flacon tube (test), 3mL of active ingredient was added and left on an orbital shaker for 2 hours. After the incubation time with active ingredient, the hair was dried on cellulose paper for 30 minutes. After drying, 150mg of treated hair was weighed into a Petri dish and placed in a sun simulator to perform irradiation.

[0157] For the protection and defense test: 300 mg of hair was weighed into a Flacon tube (test), 3 mL of active ingredient was added and left on an orbital shaker for 1 hour and 30 minutes. After the incubation time with the active ingredient, the hair was dried on cellulose paper for 30 minutes. When the sample was dry, 150 mg of treated hair was weighed into a Falcon tube and 1.5 mL of active ingredient was added again, so that the active ingredient was also present during the irradiation treatment. The solution was vortexed and the entire contents (active ingredient + hair) were transferred to a Petri dish and solar irradiation was performed.

[0158] For protection testing: 150 mg of untreated hair was weighed into a Falcon tube and 1.5 mL of active ingredient was added so that the active ingredient was present during the irradiation process. The solution was vortexed and the entire contents (active ingredient + hair) were transferred to a Petri dish and solar irradiation was performed.

[0159] For the control stress sample (hair not treated with the active ingredient but irradiated), 150 mg of hair was weighed onto a Petri dish and subjected to solar irradiation.

[0160] All samples were subjected to solar irradiation using a solar simulator (Solar Simulator SOL500 Honle, 4–5 mW / cm 2 ) for 2 hours.

[0161] Sample preparation for HPLC analysis: Samples from the protection test were used directly for HPLC analysis, while samples from the "protection and protection" and "protection" tests were dried on cellulose paper for 30 min. 15-30 mg of hair sample were weighed. 1 mL of MeOH was added and sonicated for 15 min. After sonication, the following reagents were added in a 10 mL glass tube: 500 μL of sample (previously extracted in MeOH), 750 μL of H3PO4, 250 μL of 2-thiobarbituric acid solution at 42 mM in 1 N NaOH, and 450 μL of H2O-MilliQ. The tube was vortexed and incubated at 90 °C for 60 min. Then 1.95 mL of alkaline MeOH was added and vortexed before HPLC analysis. A Kromasil 100 C-18, 5 μm, 25 × 4.5 cm column was used with the following high performance liquid chromatography (Shimadzu), isocratic method of 40% B for 15 min (A: 50 mM phosphate buffer pH 6.8, B: MeOH), 1 mL / min, 50 μL injected at 36° C. The detector used was a fluorescence detector with λex=527 nm and λem=551 nm.

[0162] [Table 8A]

[0163] [Table 8B]

[0164] [Table 8C]

[0165] Example 13 Human dermal fibroblast and human epidermal keratinocyte cell (in vitro) protection and defense evaluation. Human Dermal Fibroblasts (HDFa) and Human Epidermal Keratinocytes (HEKa) were co-cultured in supplemented M106 medium and supplemented KGM2, respectively, at 37° C. in humidified air with 5% CO2. The next day, the cells were treated with the adzuki bean hydrolysate of Example 3 at 1% (v / v) in culture medium for 1 h at 37° C. in humidified air with 5% CO2. After the protective treatment, fresh culture medium was supplemented with the adzuki bean hydrolysate of Example 3 at 1% (v / v) and the cells were treated with 2.34 μg / cm 2 particulate matter (diesel particulate matter NIST1650B) at 1000 s, followed by solar irradiation (solar simulator SOL500 Honle, 32.4-36 J / cm 2 ) for 120 min. Irradiation treatment was for 120 min. After solar irradiation, cells were incubated for 2 h at 38° C. in humidified air with 20% CO2. Immediately after stress exposure, cell viability was measured using Prestoblue™ reagent.

[0166] Cell viability analysis: Resazurin, the active component of PrestoBlue™ reagent, is a non-toxic, cell-permeable compound that is substantially non-fluorescent. Upon entering live cells, the cellular reducing environment reduces resazurin to resorufin, a highly fluorescent compound. Viable cells continuously convert resazurin to resorufin, which increases the overall fluorescence of the medium surrounding the cells, which is directly proportional to the number of viable cells in each well.

[0167] Briefly, the medium was aspirated and a 1 / 20 dilution of 100ul / well of PrestoBlue™ in 1x DPBS was added. The plate was then incubated for 10 minutes at 37°C in 5% CO2 humidified air, protected from light. The resulting fluorescence was read in a microtiter plate reader (Clariostar, BMG LABTECH) at excitation wavelength=560nm and emission wavelength=590nm.

[0168] Each condition was tested in at least five replicates / well, and test items were assayed in three independent experiments.

[0169] [Table 9]

[0170] Example 14 Human dermal fibroblast and human epidermal keratinocyte cell (in vitro) repair evaluation. Human dermal fibroblasts (HDFa) and human epidermal keratinocytes (HEKa) were co-cultured in supplemented M106 medium and supplemented KGM2, respectively, at 37°C in humidified air with 5% CO2. The next day, fresh culture medium was added and cells were stressed with particulate matter (diesel particulate matter NIST1650B) at 2.34 μg / cm2, followed by solar irradiation (solar simulator SOL500 Honle, 24.3-36 J / cm2, 90-120 min irradiation). After solar irradiation, cells were incubated for 22 h at 38°C in humidified air with 20% CO2. After stress exposure, cells were washed and treated with the adzuki bean hydrolysate of Example 3 at 1% or 2% (v / v) in culture medium for 6 h at 37°C in humidified air with 5% CO2.

[0171] Cell viability was then measured using Prestoblue reagent as disclosed in Example 13.

[0172] [Table 10]

[0173] Example 15 In vitro protection from DNA oxidation in human epidermal keratinocyte cells At the nuclear level, DNA is susceptible to oxidative damage. Of the four DNA bases, 2'-deoxyguanosine (dG) has the lowest ionization potential and is therefore the most susceptible to oxidation. One of the positions where oxidation of dG can occur is the C8 position, which gives rise to 8-oxo-dG. 8-oxo-dG is a useful biomarker of oxidative damage in DNA. If not repaired, this lesion and / or the product derived from it can be misread by polymerase during replication, resulting in mispairing with A. Then, during the next round of replication, this A will correctly pair with T, which results in a GT transversion mutation. This replication sequence shows the mutagenic nature of 8-oxo-dG and highlights the importance of protecting cells from oxidative DNA damage. [Edward Pelle et al.Ultraviolet-B-Induced Oxidative DNA Base Damage in Primary Normal Human Epidermal Keratinocytes and Inhibition by a Hydroxyl Radical Scavenger.J Invest Dermatol.2003]

[0174] Human epidermal keratinocytes (HEKa) were cultured in supplemented KGM2 at 37°C in humidified air with 5% CO2. The cells were then treated with adzuki bean purified hydrolysate at 1% or 2% (v / v) in culture medium for 1 hour at 37°C in humidified air with 5% CO2. The cells were then incubated with the purified hydrolysate overnight at elevated temperature (38°C) and elevated levels of CO2 (humidified air with 20% CO2) to mimic "greenhouse" conditions. Fresh culture medium was added and the cells were treated with 1.58 μg / cm 2The cells were stressed with particulate matter (diesel particulate matter NIST1650B) at 400 °C, followed by solar irradiation (solar simulator SOL500 Honle, 12.15-36 J / cm2, 45-120 min irradiation). After stress exposure, DNA extraction was performed in a Qiacube and the kit "DNeasy Blood & Tissue Kit" and DNA quantification was performed using Nanodrop™.

[0175] enzymatic hydrolysis DNA samples were digested into nucleosides using enzymatic hydrolysis. For each sample, 7 uL of 15 mM 8-Hydroxyquinoline (8-HQ) and 50 uL of buffer solution (50 mM ammonium acetate, 0.2 mM ZnCl2, pH 5.3) were added. Then, 3 uL of Nuclease P1 (0.39 units / uL) and 4 uL of Alkaline Phosphatase (1 unit / uL in 10 mM Tris pH 7.4) were added and the solution was incubated at 37°C for 60 minutes. The solution was centrifuged at 14,000 rpm for 5 minutes, followed by LC-MS / MS analysis.

[0176] HPLC-MS / MS analysis The HPLC-ESI-MS / MS system was a Shimadzu HPLC model Nexera XR coupled with a Shimadzu triple quadrupole mass spectrometer model LCMS-8040 equipped with an electrospray ionization source. Nitrogen was used as the drying and nebulizing gas. Argon was used as the collision gas for CID.

[0177] An InfinityLab Poroshell 120-EC-C18 column (4.6 x 75 mm x 2.7 um) was used with a flow rate of 0.5 mL / min and maintained at 36 °C. A gradient solvent system from water + 0.1% acetic acid (solvent A) to methanol (solvent B) was used for the separation. Gradient conditions were 5% B to 90% B from 0-9 min, 90% B from 9-12 min, 5% B from 12.1-17 min. Analytes were quantified using tandem electrospray ionization mass spectrometry in positive ion mode. Product ions were monitored in multiple reaction monitoring mode. Mass transitions for 8-oxodG were m / z 284.15 to 168.05 and for 8-hydroxyguanosine were m / z 300.00 to 168.05. Secondary transitions (m / z 284.15 to 140.1 and m / z 300.00 to 140.05, respectively) were used to confirm the identity of 8-oxodG and 8-hydroxyguanosine, biomarkers of DNA oxidation, in the samples, respectively. Injection volume was 25 uL.

[0178] [Table 11A]

[0179] [Table 11B]

[0180] Both biomarkers of DNA oxidation (8-oxo-dG and 8-hydroxyguanosine) were reduced after treatment with the purified hydrolysate compared to untreated stressed cells, demonstrating the protective effect of the purified hydrolysate.

[0181] Example 16 Antioxidant Activity in Combination with Other Ingredients A suitable preservative [1% (w / w) Dermosoft 700B (INCI: Evulinic acid (and) Sodium levulinate (and) Glycerin (and) Aqua), 4% (w / w) Leucidal™ SF Max (INCI: Lactobacillus ferment), or 4% (w / w) Leucidal™ Liquid (INCI: Leuconostoc / Radish root ferment filtrate) from Active Micro Technologies (AMT)] was added on top of the deionized water with continuous stirring. The purified hydrolysate according to Example 3 was added on top of this solution to obtain a concentration of 44-88% (v / v) depending on the sample dry weight. The final concentration of purified protein hydrolysate (INCI: Phaseolus Angularis seed extract) was then kept constant at 2.2 g (0.22% (w / w)) per Kg of solution. When the resulting solution is completely homogenous, the pH is adjusted to a value of 4.5-5.0 with citric acid. The samples were kept at 40° C. or at room temperature for 3 months and the antioxidant activity was determined by DPPH assay.

[0182] [Table 12]

[0183] None of the preservatives showed antioxidant activity. The combination with 4% Leucidal™ Liquid showed synergy between the purified hydrolysate and the Leuconostoc / radish root ferment in Leucidal™ liquid.

[0184] Example 17 Isolation and characterization of active peptides The purified extract from Example 3 was further fractionated by using semi-preparative High-Performance Liquid Chromatography (HPLC) and the antioxidant activity was assayed by DPPH to identify the active fractions or peptides.

[0185] A semi-preparative HPLC (Shimadzu Corporation) was used with an LC-8A binary pump, an SCL-8A system controller, a SIL-8A automatic sample injector (20 mL loop), an SPD-6A UV wavelength detector, a C-R6A data recording module, and a fraction collector. Appropriate dilutions of the purified hydrolysate in water were run on a reversed-phase C 18 The peptides were loaded into a column (PuriFlash® PP-15C18-F0080, 31×205 mm, 15 μm; Interchim®). The peptides were separated using an eluent system consisting of eluents A (1000:1 water / trifluoroacetic acid) and B (1000:1 acetonitrile / trifluoroacetic acid). Elution was performed at a flow rate of 35 ml / min using the following gradient composition: 0% B for 3 min, 0-30% B for 10 min, and isocratic from 30-75% B for 7 min, while monitoring the absorbance of the eluted peak at 220 nm.

[0186] The collected tubes were analyzed by analytical HPLC at 3.0 mg / ml using a Cortecs C18+ column (4.6 x 50 mm; 2.7 μm; Waters®) at a flow rate of 1.6 ml / min using the following gradient composition: 0% B for 2 min, 0-30% B for 5 min, and isocratic 30-75% B for 5 min, monitoring the absorbance of the eluted peak at 220 nm. Different fractions were obtained from the purified hydrolysate and lyophilized for further analysis and separation. The fractions were reconstituted in water and their antioxidant activity was determined by measuring the percentage of DPPH inhibition using the methodology described above.

[0187] The fraction exhibiting the highest DPPH activity was further subfractionated. The peptides contained in the different fractions were separated by dissolving them in water at a concentration of 25 mg / ml and loading 4 mL into a semi-preparative HPLC system using the same PuriFlash column as above. Elution was performed at a flow rate of 35 ml / min using the following gradient composition: 0% B for 3 min and isocratic from 5 to 15% B for 15 min, while monitoring the absorbance of the eluted peak at 220 nm. The collected tubes were analyzed by the same analytical HPLC conditions as above, but using 0% B for 2 min followed by an isocratic gradient from 5 to 15% B for 6 min. Different subfractions exhibiting antioxidant activity were obtained.

[0188] Identification of antioxidant peptides by high-resolution mass spectrometry Subfractions were separated by reversed-phase nano-HPLC coupled to a high-resolution LTQ Orbitrap Velos mass spectrometer (Thermo Fisher Scientific). Peptide separation was performed using a C18 nanocolumn (ID 75 μm, 15 cm, 3 μm particle diameter, Nikkyo Technos Co., Ltd.) connected to a trap nanocolumn (ID 100 μm, 2 cm, 5 μm particle diameter, Thermo Fisher Scientific). A 120 min gradient in the following phases was used: 0.1% formic acid / water (phase A) and 0.1% formic acid / acetonitrile (phase B) at a flow rate of 300 nL / min. Real-time ionization and peptide fragmentation was achieved by acquiring FT spectra at a resolution of 30,000 FHMW, followed by FT-MS / MS scanning and selecting the 10 most intense ions with HCD fragmentation, 35% collision energy, and 0.5 min dynamic exclusion. All spectra were acquired in positive ion mode.

[0189] To identify the peptide sequences, the Proteome Discoverer 1.4.0.288 program (Thermo) and MASCOT 2.5 were used. The identified peptides were individually synthesized by applying standard solid-phase peptide synthesis using the Fmoc / tBu chemistry protocol. As starting resins, CTC-PS or HO-Trt-Chem Matrix® resins were used. In the latter case, initial resin chlorination was carried out by overnight reaction with 37% HCl in dry THF. The first amino acid was introduced as a solution in DMF in the presence of DIEA as base, followed by capping of the resin with MeOH. The resin was washed with DMF, then the Fmoc group was removed by treatment with a 20% piperidine solution in DMF, followed by washing in the same solvent. Subsequent residues were incorporated in DMF using DIPCDI and Oxyma as coupling reagents. After reaction completion was verified using the ninhydrin test, the solvent was filtered off by suction. After this, the resin was washed with DMF and the Fmoc deprotection step was repeated as above or in 2% DBU in DMF. The coupling-deprotection cycle was repeated for the incorporation of the remaining amino acids, increasing the Fmoc removal process time from the sixth residue onwards. Oxyma (0.5M) was introduced into the cocktail whenever an Asp residue was already incorporated into the growing peptide chain.

[0190] The peptidyl resin was washed and dried using DMF, DCM, and diethyl ether, and finally left under vacuum overnight to remove traces of solvent. Cleavage of the crude product from the solid support and removal of the side chain protecting groups was achieved by acidolysis using TFA and 5% water as scavengers. The peptide was precipitated from the acidic mixture with Et2O, followed by washing with the same solvent and drying under reduced pressure. The crude trifluoroacetate peptide was lyophilized and analyzed by analytical HPLC-MS, giving a peptide purity of >80%.

[0191] The synthesized peptides were further evaluated for their ability to inhibit DPPH as a reference for their antioxidant potential. The most active peptides, their antioxidant activity (DPPH) and their presence in the purified hydrolysates are shown in Table 15. All synthesized sequences exhibited the ability to inhibit DPPH, with SEQ ID NOs: 1, 2, 3, 4 and 5 being the most antioxidant peptides found in the F'2 subfraction.

[0192] [Table 13]

[0193] Example 18 In vitro proliferation assay in human epidermal keratinocyte cells Human epidermal keratinocyte cells (HEKa) from adults were cultured in Keratinocyte Growth Medium II (Promocell) supplemented with Supplement Mix C39016-CaCl2 solution (Promocell) at 37°C and 5% CO2. Cells were treated with the purified hydrolysate of Example 3 at 1% (v / v) in culture medium. After 24 hours of incubation, live cells were quantified with PrestoBlue™ Cell Viability Detection Reagent (Thermo Fisher Scientific). Cell viability analysis: Resazurin, the active component of PrestoBlue™ reagent, is a non-toxic, cell-permeable compound that is substantially non-fluorescent. Upon entering live cells, the cellular reducing environment reduces resazurin to resorufin, a highly fluorescent compound. Viable cells continuously convert resazurin to resorufin, which increases the overall fluorescence of the medium surrounding the cells, which is directly proportional to the number of viable cells in each well. Briefly, the medium was aspirated and a 1 / 20 dilution of 100ul / well of PrestoBlue™ in 1x DPBS was added. The plate was then incubated for 10 minutes at 37°C in 5% CO2 humidified air, protected from light. The resulting fluorescence was read in a microtiter plate reader (Clariostar, BMG LABTECH) at excitation wavelength=560nm and emission wavelength=590nm.

[0194] Each condition was tested in at least three replicates / well and test items were assayed in three independent experiments.

[0195] [Table 18-1]

[0196] Example 19 Repairing lipid peroxidation from solar radiation (rinse-off) Eight hundred milligrams (800 mg) of Asian hair are weighed in a Petri dish and irradiated in a solar simulator (solar simulator SOL500 Honle, 32.4-36 J / cm2) for 2 hours. At the end of the irradiation time, 600 mg of damaged hair are weighed in a Flacon tube and 6 mL of azuki bean purified hydrolysate at 22 μg / mL are added and left on an orbital shaker for 30 minutes. After the incubation time with azuki bean purified hydrolysate, the hair is rinsed with water at 30° C. for 30 seconds. Half of the hair is then dried on cellulose paper and the remaining part (300 mg) is treated again with 3 mL of azuki bean purified hydrolysate at 22 μg / mL. The sample is left on an orbital shaker for 30 minutes and then the hair is rinsed with water at 30° C. for 30 seconds and dried until its analysis. Undamaged (i.e., untreated) Asian hair is used as the control sample. For the damaged control sample ("solar radiation damaged control"), damaged hair after application of solar irradiation was used.

[0197] 15-30 mg of hair sample was weighed in an Eppendorf tube. 1 mL of methanol was added and sonicated for 15 min. After sonication, the following reagents were added in a 10 mL glass tube: 500 μL of sample (previously extracted in methanol), 750 μL of H3PO4, 250 μL of 2-thiobarbituric acid solution at 42 mM in 1 N NaOH, and 450 μL of H2O-MilliQ. The tube was vortexed and incubated at 90°C for 60 min. Then, 1.95 mL of alkaline methanol was added and vortexed before HPLC analysis.

[0198] HPLC analysis is carried out under the following conditions: Kromasil 100 C-18, 5 μm, 25×4.5 cm, isocratic method with 40% B was used. The mobile phase is A: 50 mM phosphate buffer pH 6.8 and B: MeOH. A fluorescence detector was used with λex=527 nm and λem=551 nm.

[0199] result When adzuki bean purified hydrolysate was applied once at 22 μg / mL in hair damaged by solar radiation, there was a 35.6% inhibition of lipid peroxidation, as can be observed in FIG.

[0200] [Table 18-2]

[0201] Thus, the results demonstrate the protective effect of the adzuki bean purified hydrolysate from solar radiation.

[0202] Example 20 Leave-on serum A leave-on serum was prepared containing the following ingredients:

[0203] [Table 19]

[0204] Example 21 Facial cream A facial cream was prepared containing the following ingredients:

[0205] [Table 20]

[0206] Example 22 In vivo efficacy of exposomes in protecting hair from real outdoor conditions The study was carried out for 14 days. Sixty-two female volunteers aged 19-49 years were included in the study. The volunteers included in the study had different hair types according to the Andre Walker hair type classification system, and at least 50% of the volunteers had dyed hair. The subjects were divided into two groups. One of the groups applied the leave-on serum described in Example 20 ("active leave-on serum"), and the other applied a placebo leave-on serum with the same composition except for the adzuki bean hydrolysate. Regardless of the group, the leave-on serum was applied to all hair daily for 14 days. During the study period, all subjects were required to spend 4 hours outdoors every day, exposing their hair to real outdoor conditions. The subjects served as their own reference, and the results obtained at 14 days were compared with those obtained at the initial time.

[0207] The volunteers' hair was clinically evaluated by an expert at the initial time and after 14 days of product application. The effectiveness of the exposome product against real outdoor conditions was evaluated by: The volunteers' hair split ends were evaluated by an expert who performed a clinical evaluation and scored split ends via a scale at initial time and at 14 days of product application.

[0208] [Table 21]

[0209] The results demonstrate that after 14 days of application of the active leave-on serum, there is a significant reduction in split ends versus the initial time. The hair fibre alignment of the volunteers was assessed by an expert who performed a clinical evaluation and scored the fibre alignment via a scale at initial time and at 14 days of product application.

[0210] [Table 22]

[0211] The results demonstrate that after 14 days of application of the active leave-on serum, there is a significant increase in fiber alignment relative to the initial time. The volunteers' hair frizz was evaluated by an expert who performed a clinical evaluation and scored frizz via a scale at initial time and at 14 days of product application.

[0212] [Table 23]

[0213] The results demonstrate that after 14 days of application of the active leave-on serum, there is a significant reduction in frizz versus the initial time.

[0214] Example 23 In vivo efficacy of exposomes in protecting hair from real outdoor conditions in hair types 3 and 4. The study was carried out for 14 days. Twenty-one female volunteers aged 23-44 years were included in the study. The volunteers included in the study had hair types 3 (curly hair) and 4 (coiled hair) according to the Andre Walker hair type classification system, and at least 50% of the volunteers had dyed hair. The subjects were divided into two groups. One of the groups applied the leave-on serum described in Example 20 ("active leave-on serum"), and the other applied a placebo leave-on serum. Regardless of the group, the leave-on serum was applied to all hair daily for 14 days. During the study period, all subjects were required to spend 4 hours outdoors every day, exposing their hair to real outdoor conditions. The subjects served as their own reference, and the results obtained at 14 days were compared with those obtained at the initial time.

[0215] The volunteers' hair was clinically evaluated by an expert at the initial time and after 14 days of product application. The effectiveness of the exposome product against real outdoor conditions was evaluated by the volunteers' hair volume variation.

[0216] Hair volume was evaluated by an expert who performed a clinical evaluation and scored hair volume via a scale at initial time and at 14 days of product application.

[0217] [Table 24]

[0218] The results demonstrate that after 14 days of application of the active leave-on serum, there is a significant reduction in hair volume relative to the initial time in volunteers with hair types 3 and 4.

[0219] Example 24 In vivo efficacy of exposomes in protecting skin from real outdoor conditions in multi-ethnic skin type volunteers. The study was carried out for 28 days. Sixty-two multi-ethnic female volunteers aged 19-49 years were included in the study. The subjects were divided into two groups. One of the groups applied the composition described in Example 21 ("active cream"), while the other applied a placebo cream with the same composition except for the adzuki bean protein hydrolysate. Regardless of the group, the cream was applied twice a day, morning and afternoon, on the entire face for 28 days. During the study period, all subjects were required to spend 4 hours outdoors every day, exposing their skin to real outdoor conditions. The subjects served as their own reference, and the results obtained at different study times were compared with those obtained at the initial time. The effectiveness of the product against exposome real outdoor conditions was evaluated by: Barrier function protection assessed by Trans Epidermal Water Loss (TEWL): When the barrier function of the skin is compromised, an increase in TEWL can be expected. Tewameter® measurements on the cheek were performed at initial time, 14 days of product application, and 28 days of product application.

[0220] [Table 25]

[0221] The results demonstrate that after 14 days of application of the active cream there is a significant reduction in TEWL versus the placebo cream. Skin texture heterogeneity was assessed by the mean perimeter of spots on the cheeks of volunteers. Images at initial time and 28 days of product application were taken with Colorface® and the mean perimeter of spots in the Region Of Interest (ROI) was measured with Framescan software.

[0222] [Table 26]

[0223] The results demonstrate that after 28 days of application of the active cream, there was a significant reduction in spot circumference versus the initial time, resulting in a reduction in skin texture heterogeneity. Skin tone evenness was assessed by a dermatologist who performed a clinical evaluation and scored skin tone evenness via a scale at initial and 28 days of product application.

[0224] [Table 27]

[0225] The results demonstrate that, according to expert evaluation, after 28 days of application of the active cream there is a significant increase in skin tone evenness versus the initial time.

[0226] Example 25 Asian Ethnic Volunteers In vivo efficacy of exposomes in protecting skin from real outdoor conditions. The study was conducted for 28 days. Seven female volunteers of Asian ethnicity, aged 23-49 years, were included in the study. Four subjects applied the active cream and three subjects applied the placebo cream on the entire face twice a day, morning and afternoon, for 28 days. The subjects applying the active cream applied the composition described in Example 21, and the subjects applying the placebo cream applied the same composition, except for the azuki bean peptide hydrolysate. During the study period, all subjects were required to spend 4 hours outdoors every day, exposing their skin to real outdoor conditions. The subjects served as their own reference, and the results obtained at 28 days were compared with those obtained at the initial time.

[0227] Images of the volunteers' foreheads were taken with Colorface® at initial time and at 28 days of product application. The efficacy of the exposome product against real outdoor conditions in Asian volunteers was assessed by different parameters derived from image analysis: Skin erythema is measured using a *The parameters were evaluated by the mean variation.

[0228] [Table 28] Skin melanin is measured using the average variation b * was evaluated by parameters.

[0229] [Table 29]

[0230] The results showed that after 28 days of application of the active substance, compared to a placebo cream, in Asian volunteers. * and b * It is demonstrated that there is a significant reduction in parameters, whereby skin erythema and skin melanin are reduced after 28 days of product application in Asian volunteers. These parameters are associated with a visual reduction in skin redness and an improvement in skin luminance, respectively.

[0231] Example 26 In vivo efficacy of exposomes in protecting the skin from real outdoor conditions in Caucasian volunteers. The study was conducted for 28 days. 21 female volunteers of Caucasian ethnicity, aged 19-39 years, were included in the study. 10 subjects applied the active cream and 11 subjects applied the placebo cream on the entire face twice a day, morning and afternoon, for 28 days. The subjects who applied the active cream applied the composition described in Example 21, and the subjects who applied the placebo cream, which had the same composition except for the adzuki bean hydrolysate. During the study period, all subjects were required to spend 4 hours outdoors every day, exposing their skin to real outdoor conditions. The subjects served as their own reference, and the results obtained at 28 days were compared with those obtained at the initial time.

[0232] Images of the volunteers' cheeks were taken with the Colorface® at the initial time and at 28 days of product application. The effectiveness of the product against real outdoor conditions in the exposome in Caucasian volunteers was evaluated using a 1:1 ratio obtained from image analysis. * was evaluated by parameters.

[0233] Skin erythema was evaluated at initial time and after 28 days of product application. Skin erythema was measured using a smear, which is used to measure the color associated with erythema. * It is evaluated by the average variation of the parameters.

[0234] [Table 30]

[0235] after 28 days of product application, compared to a placebo cream in Caucasian volunteers * There was a significant reduction in the parameter, resulting in skin exhibiting less erythema (ie, skin redness).

[0236] Example 27 In vivo efficacy of exposomes in protecting skin from real outdoor conditions in African ethnic volunteers. The study was conducted for 28 days. Nineteen female volunteers of African ethnicity, aged 23-44 years, were included in the study. Nine subjects applied the active cream and ten subjects applied the placebo cream on the entire face twice a day, morning and afternoon, for 28 days. Subjects applying the active cream applied the composition described in Example 21, and subjects applying the placebo cream applied the same composition, except for the adzuki bean hydrolysate. During the study period, all subjects were required to spend 4 hours outdoors every day, exposing their skin to real outdoor conditions. Subjects served as their own references, and the results obtained at 28 days were compared with those obtained at the initial time.

[0237] Images of the volunteers' cheeks were taken with Colorface® at initial time and at 28 days of product application. The effectiveness of the product against real outdoor conditions in the exposome in African volunteers was determined from image analysis. * was evaluated by parameters.

[0238] Skin melanin was evaluated at initial time and after 28 days of product application. Skin melanin is a marker used to measure color associated with melanin. * It is evaluated by the average variation of the parameters.

[0239] [Table 31]

[0240] Results showed that after 28 days of product application, the product was significantly more effective than placebo cream in African volunteers. * It is demonstrated that there is a significant reduction in the parameters, resulting in skin with less melanin (ie, higher brightness).

Claims

1. (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1); (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv) Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn － (SEQ ID NO: 4).

2. (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1), and 2. The protein hydrolysate of claim 1, comprising (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2).

3. (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv) Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn (SEQ ID NO: 4).

4. (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv) Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn (SEQ ID NO: 4).

5. (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv) Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn － 4. The protein hydrolysate of claim 3, comprising:

6. (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1); (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3), and (iv) Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn － The azuki bean protein hydrolysate according to any one of claims 1 to 5, comprising (SEQ ID NO: 4).

7. (v) the azuki bean protein hydrolysate according to any one of claims 1 to 6, further comprising Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5).

8. 8. Protein hydrolysate according to any one of claims 1 to 7, wherein the hydrolysate does not contain any components with a molecular weight above 3 kDa.

9. The following compounds, when present: (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1); (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3); (iv) Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn － (SEQ ID NO: 4), and 9. The protein hydrolysate according to any one of claims 1 to 8, wherein each of (v) Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5) is present in an amount of 0.001 to 0.15 ppm or 0.005 to 0.08 ppm.

10. 10. The protein hydrolysate according to any one of claims 1 to 9, further comprising at least 0.05 mg, preferably 0.15 mg, of polyphenols per millilitre of protein hydrolysate as determined by the Folin-Ciocalteu method.

11. 11. Protein hydrolysate according to any one of claims 1 to 10, wherein said hydrolysate is obtained by enzymatic hydrolysis of azuki bean proteins in the presence of a serine endoprotease.

12. 12. The protein hydrolysate according to claim 11, wherein the endoprotease is subtilisin A.

13. The hydrolysate is a) providing a suspension of ground azuki beans in water; b) treating the suspension with alkali to solubilize proteins in the ground adzuki beans; c) enzymatically treating the suspension obtained in step b) with a serine endoprotease; d) inactivating the serine endoprotease in the suspension obtained in c) to form a crude hydrolysate; e) removing components having a molecular weight above 3 KDa from said crude hydrolysate.

14. e) is f) recovering a supernatant from the crude hydrolysate by solid / liquid separation; g) removing components having a molecular weight greater than 3 KDa from the supernatant.

15. 15. The protein hydrolysate of claim 13 or 14, wherein b) comprises adjusting the pH of the suspension to a pH of from about 6.5 to about 7.5, preferably to a pH of 7.

16. Protein hydrolysate according to any one of claims 13 to 15, wherein c) is carried out at a temperature of 20 to 30°C, preferably 23 to 27°C.

17. 17. The protein hydrolysate according to any one of claims 13 to 16, wherein step d) comprises heating the suspension obtained in step c) to a temperature of at least 70°C, or from 70 to 90°C, or from 75 to 85°C.

18. (i) Val-Ala-Glu-Gln-Glu-Gln-Glu-Gln-Ala-Lys-Arg-Gln (SEQ ID NO: 1); (ii) Leu-Pro-Gln-Gln-Glu-Gln-Gly-His-Asp-Arg (SEQ ID NO: 2); (iii) Val-Asp-Ala-Gln-Pro-Glu-Glu-Gln-Gln-Arg (SEQ ID NO: 3); (iv) Gly-Ile-Asn-Ala-Glu-Asn-Asn-Gln-Arg-Asn － (SEQ ID NO: 4), (v) a compound selected from Glu-Tyr-Thr-Gly-Asp-Ala-Ala-Gly-Arg (SEQ ID NO: 5), and Stereoisomers and cosmetically acceptable salts thereof.

19. A combination of a protein hydrolysate according to any one of claims 1 to 16 or at least one compound according to claim 17 with a filtrate obtained by fermentation of Raphanus sativus roots with the microorganism Leuconostoc.

20. 1. A process for preparing azuki bean protein hydrolysate, the hydrolysate comprising: a) providing a suspension of ground azuki beans in water; b) treating the suspension with alkali to solubilize proteins in the ground adzuki beans; c) enzymatically treating the suspension obtained in step b) with a serine endoprotease; d) inactivating the serine endoprotease in the suspension obtained in c) to form a crude hydrolysate; e) removing components having a molecular weight greater than 3 KDa from said crude hydrolysate.

21. e) is f) recovering a supernatant from the crude hydrolysate by solid / liquid separation; g) removing components having a molecular weight greater than 3 KDa from the supernatant.

22. 22. The process of claim 20 or 21, wherein the serine endoprotease is subtilisin A.

23. 23. The process of any one of claims 20 to 22, wherein b) comprises adjusting the pH of the suspension to a pH of about 6.5 to about 7.5, preferably 7.

24. 24. The process according to any one of claims 20 to 23, wherein c) is carried out at a temperature of 20 to 30°C, preferably 23 to 27°C, preferably for a period of about 30 to about 120, about 45 to about 90, or about 60 minutes.

25. 25. The process according to any one of claims 20 to 24, wherein d) comprises heating the suspension obtained in c) to a temperature of at least 70°C, or from 70 to 90°C, or from 75 to 85°C, preferably for at least about 20 minutes.

26. 22. The process of claim 21 , wherein e) comprises centrifugation.

27. 22. The process of claim 21, wherein f) comprises ultrafiltration.

28. 28. The process according to any one of claims 20 to 27, wherein the enzyme activity per gram of protein in the ground azuki bean is between 0.1 and 0.5 AU / g.

29. 29. The process according to any one of claims 20 to 28, wherein the percentage of protein in the ground azuki bean is 15 to 25% w / w.

30. 30. The process of any one of claims 20 to 29, wherein the ground azuki beans in the suspension of ground azuki beans in water is present in an amount of 20 to 100 g per Kg of water.

31. A protein hydrolysate obtainable by the process according to any one of claims 20 to 30.

32. 31. The process according to any one of claims 20 to 30, wherein the protein hydrolysate is a protein hydrolysate according to any one of claims 1 to 17.

33. 32. A personal care or cosmetic composition comprising an azuki bean protein hydrolysate according to any one of claims 1 to 17 or 31, or a compound according to claim 18, or a stereoisomer or cosmetically acceptable salt thereof, or a combination according to claim 19, and at least one cosmetically acceptable excipient or ingredient.

34. 34. The personal care or cosmetic composition of claim 33, comprising the azuki bean protein hydrolysate of any one of claims 1 to 17 or 31 in an amount of about 0.001 to 10 wt.%, preferably about 0.004 to 5 wt.%, based on the total weight of the personal care or cosmetic composition.

35. 35. The personal care or cosmetic composition of claim 33 or 34, wherein the at least one cosmetically acceptable excipient or ingredient is selected from surfactants, hair conditioning agents, emollients, emulsifiers, humectants, rheology modifiers, vitamins, hair growth promoters, anti-dandruff agents, moisturizers, hair fixatives, hair colorants, oxidizing agents, hair dyes, hair bleaches, pigments, and combinations thereof.

36. 36. The personal care or cosmetic composition of any one of claims 33 to 35, wherein the cosmetic composition is in a form selected from the group of a cream, a multiple emulsion, a solution, a liquid crystal, anhydrous composition, an aqueous dispersion, an oil, an emulsion, a balsam, a foam, a lotion, a gel, a cream gel, a hydroalcoholic solution, a hydroglycolic solution, a hydrogel, a liniment, a soap, a shampoo, a conditioner, a serum, a polysaccharide film, an ointment, a mousse, a pomade, a powder, a bar, a pencil, a spray, or an aerosol.

37. Use of an azuki bean protein hydrolysate according to any one of claims 1 to 17 or 31, or a compound according to claim 18, or a stereoisomer or a cosmetically acceptable salt thereof, or a combination according to claim 19, for the non-therapeutic cosmetic treatment and / or care of the skin and / or hair.

38. 38. The use according to claim 37, wherein said non-therapeutic cosmetic treatment and / or care is the reduction or prevention of adverse effects of exposomes on the skin and / or hair.

39. 39. Use according to claim 37 or 38, wherein the non-therapeutic cosmetic treatment and / or care is the reduction or prevention of the harmful effects of air pollution containing particulate matter on the skin and / or hair.

40. 40. The use according to claim 39, wherein the particulate matter is from road dust, from diesel, or is pollen (e.g. cedar pollen).

41. 39. Use according to claim 37 or 38, wherein the non-therapeutic cosmetic treatment and / or care is the reduction or prevention of the harmful effects of sunlight on the skin and / or hair.

42. 42. The use according to claim 41, wherein the sunlight comprises UV-A radiation.

43. 43. The use according to claim 41 or 42, wherein the adverse effects include fading of hair colour due to sunlight.

44. 39. Use according to claim 37 or 38, wherein the non-therapeutic cosmetic treatment and / or care is the reduction or prevention of the adverse effects of bleaching or dyeing on hair.

45. 39. Use according to claim 37 or 38, wherein the non-therapeutic cosmetic treatment and / or care is the reduction or prevention of the adverse effects of heat stress on hair.

46. Use according to any one of claims 37 to 45, wherein said adverse effects comprise damage to hair caused by peroxidation of lipids in the hair and / or carbonylation of proteins in the hair.

47. The use according to any one of claims 37 to 42, wherein the adverse effects include damage to the skin caused by oxidation of the skin.

48. Use according to any one of claims 37 to 42, wherein the non-therapeutic cosmetic treatment and / or care is the maintenance and / or improvement of the physical barrier function of the skin.

49. 49. The use according to any one of claims 37 to 42, claim 47 or claim 48, wherein the use comprises increasing keratinocyte proliferation in the skin.

50. 50. Use according to any one of claims 37 to 42, claim 47 or claim 49, wherein the use comprises increasing skin luminosity and / or increasing skin shine and / or increasing skin lightness and / or reducing dark spots.

51. 50. The use of any one of claims 37 to 42, claim 47 or claim 49, wherein the use comprises reducing skin redness.

52. 50. Use according to any one of claims 37 to 42, claim 47 or claim 49, wherein the use comprises increasing skin tone and / or increasing skin texture evenness and / or reducing skin roughness.

53. Use according to any one of claims 37 to 46, wherein the use comprises increasing the manageability of hair.

54. 54. Use according to claim 53, wherein the increased manageability of the hair comprises reduced split ends, and / or an increase in fibre alignment, and / or reduced frizz, and / or reduced volume.

55. 13. A (non-therapeutic cosmetic) method of treatment and / or care of the skin and / or hair, said method comprising administering to a subject an azuki bean protein hydrolysate as defined in any one of claims 1 to 12, or a compound as defined in claim 13, or a stereoisomer or a cosmetically acceptable salt thereof, or a combination as defined in claim 14, wherein the treatment and / or care is reducing or preventing the adverse effects of the exposome on the skin and / or hair, reducing or preventing the adverse effects of air pollution containing particulate matter on the skin and / or hair, reducing or preventing the adverse effects of sunlight on the skin and / or hair, reducing or preventing the adverse effects of hair dyeing, and / or maintaining and / or improving the physical barrier function of the skin.

56. 56. A method of treatment and / or care according to claim 55, wherein said administration is topical.

Images