A skin care composition

Abstract

The present invention is in the fields of skin care compositions. Particularly, it relates to a skin care composition delivering antimicrobial benefit in 'eco-friendly' manner. There is a need for an improved skin care composition for providing antimicrobial benefit using environmentally sustainable ingredients. Surprisingly it has been found that a skin care composition comprising a gamma-keto carboxylic acid, or its salt or its ester along with select antimicrobial peptide (AMP) and / or their boosters enhances AMP secretion and / or its functionality, thereby providing enhanced antimicrobial benefit in a sustainable manner.

Description

[0001] P0000689 COM

[0002] 1

[0003] A SKIN CARE COMPOSITION

[0004] Field of the invention

[0005] The present invention is in the field of skin care compositions. Particularly, it relates to a skin care composition delivering antimicrobial benefits using environmentally sustainable ingredients.

[0006] Background of the invention

[0007] Consumers tend to take care of themselves to maintain a good health. This includes taking caring of their body parts, particularly skin, axilla, oral cavity, hair and scalp. Consumers are often looking for infection-free skin, free of body odour, better skin tone and moisturized skin. Skin is the largest body part of a human, it serves as the primary line of defence from invading microorganism, such as, virus, fungi and bacteria and prevent infection. Often skin contains commensal bacteria which do not cause infection but rather help in preventing pathogenic or non-commensal bacteria from taking hold. Apart from this, skin has its own defence mechanism, such as, anti-microbial peptide (AMP), which also helps in fighting with pathogenic or non-commensal bacteria.

[0008] Conventionally, skin care compositions contain antimicrobial agents for providing broad spectrum antimicrobial benefit. However, consumers are nowadays concerned about sustainability and the environment, and they prefer to have cleaning products with a good environmental profile. That is, they prefer products that contain less chemicals or which have ‘eco-friendly’ ingredients and have less or no impact on the environment when the product is made or used.

[0009] In this context, strengthening skin’s own defence mechanism through topical application is one of the ways to move in this direction. Antimicrobial peptides (AMP) are integral part of the skin’s own defence mechanism. AMPs are ubiquitous in nature, and they typically exhibit a broad spectrum of activity against invading bacteria, fungi, viruses and parasites. They are generally short peptides. Reportedly there are about 90 different AMPs found in humans. In general, they have two major physical features, namely cationic charge and a significant proportion of hydrophobic residues. The cationic charge of the AMPs promotes selectivity for negatively charged microbial surfaces whereas the hydrophobicity facilitates interactions with the cell membrane of the microbial species.P0000689 COM

[0010] 2

[0011] Enhancing AMP secretion and / or its functionality is one of the steps in delivering antimicrobial benefit in an ‘eco-friendly’ manner.

[0012] CN105944076A discloses an auristilla used for treating chronic suppurative otitis media containing cantharidin 1.2-1.8%, benzoic acid 0.8-1.6%, diclofenac sodium 0.6-1.0%, chrysin 0.5-0.9%, calcium phosphate 1.0-1.4%, antimicrobial peptide 0.5-0.8%, levulinic acid 0.2-0.4%, dipterocarpol 0.05-0.15%, isopropyl myristate 1.3-1.5%, vitamin C 0.3-0.5%, polyvinylpyrrolidone 0.15-0.35%, acetate buffer solution 1.5-3.5%, stabilizer 0.1-0.4%, tackifier 2.4-4.8%, pH modifier 0.5-2.5%, and the balance water for injection.

[0013] WO 2016 / 030609 A1 discloses an antimicrobial composition comprising metal silver, characterised in that it also comprises an antimicrobial peptide, and optionally a fluorinated polymer.

[0014] FR2979522A1 discloses a preservative system comprises an antimicrobial peptide, polyol, preferably propanediol, and anisic acid and a cosmetic composition containing the preservative system.

[0015] Jiang Weiyu et. al. (Peptides 50 (2013) 129-138) reports differential regulation of human cathelicidin LL-37 by free fatty acids and their analogues.

[0016] In this regard there is a need for an improved skin care composition providing antimicrobial benefit using environmentally sustainable ingredients.

[0017] Surprisingly, the present inventors have found that a skin care composition comprising a gamma-keto carboxylic acid, or it’s salt or its ester along with select antimicrobial peptides and / or their boosters enhances AMP secretions and / or its functionality; thereby it provides enhanced antimicrobial benefit in a sustainable manner.

[0018] Summary of the invention

[0019] Accordingly, in a first aspect the present invention provides a skin care composition comprising:P0000689 COM

[0020] 3

[0021] a. 0.000001 to 10% by weight of one or more compounds of formula I or II or III:

[0022]

[0023] where:

[0024] R1 is selected from H, a straight or branched C1 to C6 alkyl group or C2 to C6 allyl group, an aryl group, Na+, K+, Ca2+, Mg2+, NH4+;

[0025] R2 is selected from H, a straight or branched C1 to C14 alkyl group, an ethylamine group (CH2NH2) for formulae I or II;

[0026] R2 is H or a straight or branched C1 to C4 alkyl group for formula III; and R3 is selected from Cl Br, I’ and NH2; and

[0027] b. an antimicrobial peptide selected from defensins, histatins, cathelicidin, dermcidin, S100 peptides, RNase peptides and combinations thereof and / or a booster thereof selected from hydroxy-stearic acid, nicotinamide, isomer of nicotinamide, cycloalkyl derivatives of nicotinamide, Vitamin A, Vitamin C and linseed oil and combinations thereof, wherein the composition is free of cantharidin.

[0028] In a second aspect, the present invention relates to a non-therapeutic method for enhancing antimicrobial peptides (AMP) secretion on an external surface of the human body comprising steps of:

[0029] a. applying topically a composition according to the first aspect on to the surface;

[0030] and

[0031] b. optionally, rinsing the surface.P0000689 COM

[0032] 4

[0033] In a third aspect, the present invention relates to a use of a combination of one or more of the compounds of formula I or II or III according to the first aspect and a booster of antimicrobial peptides (AMP) selected from hydroxy-stearic acid, nicotinamide, isomer of nicotinamide, cycloalkyl derivatives of nicotinamide, Vitamin A, Vitamin C and linseed oil, and combinations thereof for enhancing secretion of antimicrobial peptides (AMP) when applied on external surfaces of the human body.

[0034] Detailed description of the invention

[0035] Any feature of one aspect of the present invention may be utilized in any other aspect of the invention. The word “comprising” is intended to mean “including” but not necessarily “consisting of” or “composed of.” In other words, the listed steps or options need not be exhaustive. Except in the operating and comparative examples, or where otherwise explicitly indicated, all numbers in this description indicating amounts of material or conditions of reaction, physical properties of materials and / or use are to be understood as modified by the word “about”. Numerical ranges expressed in the format “x to y” are understood to include x and y. When for a specific feature multiple preferred ranges are described in the format “x to y”, it is understood that all ranges combining the different endpoints are also contemplated. Unless specified otherwise, amounts as used herein are expressed in percentage by weight based on total weight of the composition and may be abbreviated as “wt%”. The use of any and all examples or exemplary language, e.g. “such as” provided herein is intended merely to better illuminate the invention and does not in any way limit the scope of the invention otherwise claimed. Except in the examples, or where otherwise explicitly indicated, all numbers in this description indicating amounts of material or conditions of reaction, physical properties of materials and / or use may optionally be understood as modified by the word “about”.

[0036] The composition according to the present invention is meant to be used for skin care or topical cosmetic use. By “skin care composition” as used herein, is meant to include a composition for topical application on to skin including axilla, hair including scalp, oral cavity. Such skin care composition may be classified as leave-on or wash-off, and includes any product applied to a human body for improving appearance, cleansing, odour control or general aesthetics. The composition can be in the form of a liquid, lotion, cream, foam, scrub, gel, soap bar or toner, or applied with an implement or via a face mask, pad, wipe, or patch. Non-limiting examples of such compositions include leave-P0000689 COM

[0037] 5

[0038] on products, like skin lotions and creams, antiperspirants, deodorants, depilatories, lipsticks, foundations, mascara, sunless tanners, sanitizers, gels, sprays or sunscreen lotions; wash-off products like shampoos, conditioners, shower gels, toilet bars, face wash, hand wash, or body wash products.

[0039] “Skin” as used herein, preferably means to include skin on any part of the body, e.g. face, neck, chest, back, arms, underarms, hands, legs, buttocks, and scalp. Compositions of the invention is also of relevance to applications on any other keratinous substrates of the human body other than skin, e.g. hair.

[0040] In a first aspect, the present invention provides a skin care composition comprising one or more compounds of formula I or II or III and an antimicrobial peptide and / or a booster thereof.

[0041] One or more compounds of formula I or II or III

[0042] The composition comprises one or more compounds of formula I or II or III:

[0043]

[0044] where,

[0045] R1 is selected from H, a straight or branched alkyl group of 1 to 6 carbon atoms, an allyl group of C2 to C6, an aryl group, Na+, K+, Ca2+, Mg2+, NH4+,

[0046] R2 is selected from H, a straight or branched alkyl group of 1 to 14 carbon atoms, an ethylamine group (CH2NH2) for formulae I or II; orP0000689 COM

[0047] 6

[0048] R2 is selected from only from either H or a straight or branched alkyl group of 1 to 4 carbon atoms for formula III; and

[0049] R3 is selected from Cl Br, I’ and NH2.

[0050] It implies that the compounds of formula I or II or III are either a carboxylic acid, or a salt, or an ester or a derivative thereof, which have 4 to 18 carbons in total; and which have either a keto group (=0) or a hydroxy group (-0H) or a halo group (any one of Cl’ or BR’ or I preferably Cl’) or a -NH2 group attached at the gamma carbon which is the 4thcarbon.

[0051] In carboxylic acids, the carbon atoms except the carbonyl carbon (which is the first carbon in the carbon chain) are often named using Greek symbols. In other words, the carbon atom adjacent to the carbonyl carbon (the first carbon atom of the carbon chain), is named with the Greek letter ‘a’ (alpha) and is known as a carbon, the next carbon atom is denoted as ‘P’(beta) carbon and the further next one as ‘Y’(gamma) carbon, and so on. A general sketch of a carboxylic acid that shows the carbonyl carbon, alpha, beta, gamma and subsequent carbons is as follows:

[0052]

[0053] An example of a compound of formula I is levulinic acid. Examples of a salt of levulinic acid include sodium levulinate, potassium levulinate, calcium levulinate, ammonium levulinate, and mixtures thereof. Examples of an ester of levulinic acid include methyl levulinate, ethyl levulinate, butyl levulinate, benzyl levulinate, allyl levulinate, and mixtures thereof.

[0054] Examples of the compound of formula I, II or II suitable for the present invention include 4-keto compounds (formula I), e.g. 4-oxo butanoic acid, levulinic acid, 4-keto n-caprinoic acid, 4-oxo octanoic acid, 4-oxo decanoic acid, 4-keto lauric acid, 4-keto myristic acid, 4-keto palmitic acid, and 4-keto stearic acid; 4-hydroxy compounds (formula II), e.g. 4-hydroxy butanoic acid, 4-hydroxy pentanoic acid, 4-hydroxy hexanoic acid, 4-hydroxy octanoic acid, 4-hydroxy decanoic acid, 4-hydroxy lauric acid, 4-hydroxy palmitic acid,P0000689 COM

[0055] 7

[0056] 4-hydroxy stearic acid; 4-chloro compounds (formula III), e.g. 4-chloro butanoic acid, 4-chloro pentanoic acid, 4-chloro octanoic acid; 4-amino compounds (formula III), e.g. 4-amino butanoic acid (GABA), 4-amino pentanoic acid, 4-amino hexanoic acid, and 4-amino octanoic acid. Preferably the compound of formula I, II or III is selected from levulinic acid, propyl levulinate, benzyl levulinate, butyl levulinate, methyl levulinate, ethyl levulinate, and combinations thereof. Preferably, the composition comprises one or more compounds of formula I or II.

[0057] The composition comprises one or more compounds of formula I, II, or II. As such a composition which does not comprise one or more compounds of formula I is envisaged (i.e. wherein the amount of compounds of formula I is 0 wt%). Similarly, a composition which does not comprise one or more compounds of formula II is envisaged (i.e. wherein the amount of compounds of formula II is 0 wt%), as is a composition which does not comprise one or more compounds of formula III (i.e. wherein the amount of compounds of formula III is 0 wt%).

[0058] The composition comprises from 0.000001 to 10% by weight, preferably from 0.00001 to 9% by weight, more preferably from 0.0001 to 8% by weight, even more preferably from 0.001 to 7% by weight, further more preferably from 0.01 to 6% by weight, yet more preferably from 0.01 to 5% by weight, still more preferably from 0.01 to 4 % by weight, most preferably from 0.01 to 3% by weight of the compound of formula I, II or III.

[0059] Antimicrobial peptide and / or a booster thereof

[0060] The composition according to the present invention comprises an anti-microbial peptide (AMP) and / or booster thereof. This implies the composition comprises AMP ora booster thereof or both.

[0061] The AMP is selected from defensins, histatins, cathelicidin, dermcidin, S100 peptides, RNase peptides, and combinations thereof. Defensins herein include human beta defensin 1 , human beta defensin 2, human beta defensin 3, and human beta defensin 4. Histatins include histatin 5 and histatin 7. LL-37 is an example of cathelicidin. Dermcidin 1 is an example of dermcidin. Psoriasin is an example of S100 peptides. RNase 7 is an example of RNAse peptides. Such AMPs may be used in the present invention individually or as a combination. More preferably, AMPs are selected from human beta defensin 1, human beta defensin 2, human beta defensin 3, human beta defensin 4, LL-P0000689 COM

[0062] 8

[0063] 37, and mixtures thereof. The antimicrobial peptide is preferably selected from defensins, histatins, S100 peptides, RNase peptides, and combinations thereof. Most preferably the AMP is selected is human beta defensin 3, LL-37, and mixtures thereof.

[0064] Preferably, the composition comprises AMPs in an amount ranging from 0.000001 to 2% by weight, more preferably from 0.00005 to 1% by weight, even more preferably from 0.00001 to 0.5% by weight, furthermore preferably from 0.0001 to 0.2% by weight and still more preferably from 0.001 to 0.1% by weight.

[0065] The anti-microbial peptide (AMP) booster is selected from one or more of hydroxy stearic acid, nicotinamide, isomer of nicotinamide, cycloalkyl derivatives of nicotinamide, Vitamin A, Vitamin C, and linseed oil, and combinations thereof. An AMP booster as per this invention is meant a compound which when applied on to the skin of a living being (e.g. human being) produces enhanced levels of AMPs on skin.

[0066] It is preferred that the hydroxy stearic acid is 10-hydroxystearic acid, 12-hydroxystearic acid or tri-hydroxy stearic acid (e.g., 9,10,13-trihydroxystearic acid) or tri-hydroxy stearin or compounds that yield one or more molecules of hydroxy stearic acid or hydroxy stearate on their breakdown like mono, di or tri ester of glycerol with hydroxy stearic acid. Of these compounds, 10-hydroxy stearic acid, 12-hydroxy stearic acid and 9,10,13-tri-hydroxy stearic acid are more preferred. Most preferred AMP booster is 12-hydroxy stearic acid (12-HSA) is most preferred, the structure of which is given below:

[0067]

[0068] Another preferred AMP booster is nicotinamide, also known as niacinamide, which is a Vitamin B3 compound. Other vitamin B3 compounds which may be used are niacin or nicotinic acid. Isomers of nicotinamide which may be used include iso-nicotinamide or picolinamide. Cycloalkyl derivatives of nicotinamide which may be used preferably have 4 to 6 carbon atoms in the cycloalkyl group, e.g. n-cyclo-butyl nicotinamide, n-cyclo-pentyl nicotinamide, and n-cyclo-hexyl nicotinamide. The most preferred Vitamin B3 compound is niacinamide. Niacinamide also known as pyridine-3-carboxamide, it is water soluble form of vitamin B3.P0000689 COM

[0069] 9

[0070] Vitamin C or its derivatives may also be used as an AMP booster in the composition of the invention. Vitamin C or its derivative for use may be selected from one or more of ascorbic acid, sodium ascorbyl phosphate, 3-0-ethyl ascorbic acid, ascorbyl methylsilanol pectinate, magnesium ascorbyl phosphate, ascorbyl glucoside, and tetrahexyldecyl ascorbate; preferably ascorbic acid or sodium ascorbyl phosphate.

[0071] The composition of the present invention may comprise a Vitamin A compound as the AMP booster, which is also known as a retinoid. Typically, the retinoid is selected from retinyl ester, retinol, retinal, retinoic acid, or a mixture thereof. More preferably the retinoid comprises retinol, retinyl ester, or a mixture thereof, and even more preferably the retinoid is selected from retinol, retinyl ester, or a mixture thereof. Of these, retinyl esters are preferred. Suitable retinyl esters for use in the present invention are retinyl palmitate, retinyl propionate, retinyl acetate, retinyl linoleate, retinyl oleate, or a mixture thereof. Most preferred are retinyl palmitate, retinyl acetate, retinyl propionate, or a mixture thereof.

[0072] Preferably the AMP booster is selected from hydroxy-stearic acid, nicotinamide, linseed oil, and combinations thereof.

[0073] The composition of the invention preferably comprises 0.00001 to 5% by weight, more preferably 0.00005 to 3% by weight and most preferably 0.0001 to 1% by weight of the AMP booster.

[0074] The composition preferably comprises a cosmetically acceptable base, which often acts as vehicle for delivering active ingredients. The cosmetically acceptable base may be a leave-on or wash-off and may include ingredients suitable for cosmetic application, such as: anionic surfactant, non-ionic surfactant, cationic surfactant, amphoteric surfactant, emollient, non-aqueous solvent, water, moisturizing agent, perfume, and colourant.

[0075] The composition is suitable for skin care applications and free of ingredients which may causes or harm to skin, such as skin irritation, redness etc. One of the examples of such ingredient is cantharidin. It is reported that cantharidin often causes skin redness, swelling, warmth, eye irritation and ulceration. The composition is free of cantharidin. The term ‘free off’ herein implies the composition does not comprise more than 0.1% byP0000689 COM

[0076] 10

[0077] weight, or preferably not more than 0.05% by weight and more preferably not more than 0.01% by weight. Most preferably the composition contains no cantharidin at all.

[0078] Leave-on composition

[0079] The composition according to the present invention preferably is delivered as a leave-on composition. The cosmetically acceptable vehicle for a leave-on composition is preferably in the form of an oil, liquid, stick, cream, lotion, spray, or gel. The cosmetically acceptable vehicle in leave-on compositions preferably includes ingredients like a nonionic surfactant, a fatty acid, a soap, a polymer, an emollient, a polyhydric alcohol, a solvent, a powder, or combinations thereof. Details on the various possible ingredients for inclusion in the cosmetically acceptable vehicle are given below.

[0080] Preferably, the composition comprises a non-ionic surfactant. More preferably the nonionic surfactant is selected from those having HLB value in the range 9 to 20. HLB is calculated using the Griffin method wherein HLB = 20 x Mh / M wherein Mh is the molecular mass of the hydrophilic portion of the molecule and M is the molecular mass of the whole molecule, giving a result on an arbitrary scale of 0 to 20. Preferably, the non-ionic surfactant having HLB value in the range 9 to 20 is selected from fatty alcohol ethoxylates, alkyl phenol ethoxylates, polyoxyethylene sorbitan alkyl esters and mixtures thereof. Another preferred non-ionic surfactant is cocamide monoethanolamide (CMEA). Preferably, the leave-on composition comprises 0.5 to 5% by weight, more preferably 1 to 4% by weight, even more preferably from 2 to 3% by weight non-ionic surfactant having HLB in the range 9 to 20.

[0081] The composition may further comprise anionic surfactant. Examples of anionic surfactants suitable for the invention include alkyl ether sulphate and sulfonates, alkyl sulphate and sulphonate, alkylbenzene sulphonates, alkyl and di-alkyl sulphosuccinate, C8 to C20 acyl isethionate, 08-020 alkyl ether phosphate, alkyl ether carboxylate, and combinations thereof.

[0082] The composition may contain a cationic surfactant. Examples of suitable cationic surfactant include palmitamidopropyltrimonium chloride, di-stearyl dimonium chloride and mixtures thereof. Useful amphoteric surfactants which may be added in the composition include cocoamidopropyl betaine, C12-C20 trialkyl betaines, sodium lauroamphoacetate, and sodium laurodiamphoacetate, or a mixture thereof.P0000689 COM

[0083] 11

[0084] Preferably, the composition according to the present invention is delivered in the form of a vanishing cream. A vanishing cream is one which when applied and rubbed on to the human skin, vanishes on the skin leaving behind no significant streaks of the composition. Fatty acids when present in a composition along with a soap provides the so-called vanishing cream effect. Preferably, the leave-on composition comprises fatty acids having 10 to 30, more preferably 12 to 25, even more preferably 14 to 20, furthermore preferably 16 to 18 carbon atoms. Examples of fatty acids that may be used in the composition include pelargonic, lauric, myristic, palmitic, stearic, iso-stearic, oleic, linoleic, arachidic, behenic, erucic acid, and mixtures thereof. Preferably, the fatty acid that may be used is stearic acid or palmitic acid or a mixture thereof. The fatty acid in the present invention is preferably hysteric acid which is substantially (generally about 90 to 95% by weight) a mixture of stearic acid and palmitic acid in a ratio of between 55:45 to 45:55.

[0085] Preferably, the vanishing cream composition comprises from 2.25 to 25 % by weight, more preferably from 4 to 22 % by weight, even more preferably from 6 to 20 % by weight, furthermore preferably from 8 to 19 % by weight and still more preferably from 10 to 18 % by weight and yet more preferably from 12 to 16 % by weight of the fatty acid.

[0086] Preferably, the vanishing cream composition comprises soap. Soap, when present in combination with fatty acid in the composition, provides the vanishing effect described above. Preferably, soap in the composition is generally prepared by in-situ neutralization of fatty acid that may be present in the composition. Thus, it is preferred that the soap has a carbon chain length that corresponds to the chain length of fatty acid in the composition. The soap is formed from the fatty acid through use of alkali metal hydroxides, e.g. sodium hydroxide or potassium hydroxide. Potassium hydroxide is more preferred. Thus, the soap is preferably a potassium soap (potassium salt of fatty acid). Preferably, the vanishing cream composition comprises from 0.1 to 10% by weight, more preferably from 0.25 to 8% by weight, even more preferably from 0.5 to 7% by weight, furthermore preferably from 0.5 to 5% by weight, even furthermore preferably 0.5 to 3% by weight of soap.

[0087] Preferably, the leave-on composition may comprise a polymer. The polymer acts as thickener in the composition and improves sensorial properties of the composition. The polymer is preferably selected from the following classes:P0000689 COM

[0088] 12

[0089] - acrylate I R-methacrylate copolymer e.g. acrylates / steareth-20 methacrylate copolymer (commercially available as Aculyn™ 22) and acrylates / beheneth-25 methacrylate copolymer (commercially available as Aculyn™ 28),

[0090] - acrylate I R-methacrylate crosspolymer e.g. acrylates / steareth-20 methacrylate crosspolymer (commercially available as Aculyn™ 88),

[0091] - acrylates copolymer (commercially available as Aculyn™ 33),

[0092] - acrylate / R-alkyl acrylate crosspolymer e.g. acrylates / C10-C30 alkyl acrylate crosspolymer (commercially available as Pemulen™ TR-2),

[0093] - copolymer of ammonium acryloyldimethyltaurate with vinyl pyrrolidone (commercially available as Aristoflex® AVC),

[0094] - copolymer of sodium acryloyldimethyltaurate with vinyl pyrrolidone (commercially available as Aristoflex® AVS); and

[0095] - crosspolymer of acryloyldimethyltaurate with R-alkyl acrylate and methyacrylate e.g. Ammonium acryloyldimethyltaurate / beheneth-25 methacrylate crosspolymer (commercially available as Aristoflex® HMB and Aristoflex® BLV).

[0096] Preferably, the composition comprises 0.1 to 5% by weight, more preferably 0.25 to 4.5% by weight, even more preferably 0.5 to 4% by weight, furthermore preferably from 0.75 to 3.5% by weight, still more preferably from 0.75 to 2.75 % by weight of the polymer.

[0097] Preferably, the composition comprises emollients. Examples of emollients that may be used in the leave-on composition include stearyl alcohol, glyceryl mono-ricinoleate, mink oil, isopropyl iso-stearate, isobutyl palmitate, iso-cetyl stearate, oleyl alcohol, isopropyl laurate, hexyl laurate, decyl oleate, octadecan-2-ol, iso-cetyl alcohol, eicosanyl alcohol, behenyl alcohol, cetyl palmitate, silicone oils such as dimethylpolysiloxane, din-butyl sebacate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, butyl stearate, polyethylene glycol, triethylene glycol, lanolin, cocoa butter, corn oil, cotton seed oil, olive oil, palm kernel oil, rape seed oil, safflower seed oil, evening primrose oil, soybean oil, sunflower seed oil, avocado oil, sesame seed oil, coconut oil, arachis oil, castor oil, acetylated lanolin alcohols, petroleum jelly, mineral oil, butyl myristate, isopropyl linoleate, lauryl lactate, myristyl lactate, decyl oleate, myristyl myristate, caprylyl triglyceride, and mixtures thereof.

[0098] Preferably the composition may comprise a polyhydric alcohol. Polyhydric alcohols are selected from one or more of glycerol, propylene glycol, dipropylene glycol,P0000689 COM

[0099] 13

[0100] polypropylene glycol, polyethylene glycol, sorbitol, hydroxypropyl sorbitol, hexylene glycol, 1,3-butylene glycol, isoprene glycol, 1,2,6-hexanetriol, ethoxylated glycerol, propoxylated glycerol, or a combination thereof. Most preferred polyhydric alcohol is glycerol.

[0101] Preferably, the composition comprises non-aqueous solvents. Examples of solvents that may be used in the composition include ethyl alcohol, isopropanol, acetone, ethylene glycol ono ethyl ether, diethylene glycol mono butyl ether, diethylene glycol mono ethyl ether, and mixtures thereof.

[0102] Preferably, the composition comprises powders. Examples of powders that may be used in the composition include zinc oxide, titanium oxide, chalk, talc, fullers earth, kaolin, starch, gums, colloidal silica sodium polyacrylate, tetra alkyl and / or trialkyl aryl ammonium smectites, chemically modified magnesium aluminium silicate, organically modified montmorillonite clay, hydrated aluminium silicate, fumed silica, carboxy vinyl polymer, sodium carboxymethyl cellulose, ethylene glycol monostearate, and mixtures thereof.

[0103] The leave-on composition may be in the form of an emulsion, e.g. a water-in-oil emulsion or an oil-in-water emulsion, preferably an oil-in-water emulsion. In such cases, the composition may include mineral oils, silicone oils, and synthetic oils. Amounts of these materials may range from 0.1 to 50% by weight, and preferably, from 0.1 to 30% by weight, and most preferably, from 1 to 20% by weight, including all ranges subsumed therein. Silicone oils may be divided into the volatile and non-volatile varieties. The term “volatile” as used herein refers to those materials which have a measurable vapor pressure at ambient temperature. Volatile silicone oils are preferably chosen from cyclic or linear polydimethylsiloxanes containing from 3 to 9, and preferably, from 4 to 5 silicon atoms. Linear volatile silicone materials generally have viscosities of less than 5 centistokes at 25°C while cyclic materials typically have viscosities of less than 10 centistokes when measured with a Brookfield Viscometer, RV No. 3 spindle at 20 PRM, standardized to mineral oil and at 25°C.

[0104] Non-volatile silicone oils useful as carrier material include polyalkyl siloxanes, polyalkylaryl siloxanes, and polyether siloxane copolymers. The essentially non-volatile polyalkyl siloxanes useful herein include, for example, polydimethylsiloxanes (likeP0000689 COM

[0105] 14

[0106] dimethicone, including cross polymers and elastomers) with viscosities of from 5 to 100,000 centistokes at 25°C. An often-preferred silicone source is a cyclopentasiloxane and dimethicone solution.

[0107] Water is present in different amounts depending on the type of leave-on composition. Liquid composition may comprise from 10 to 95 % by weight, more preferably 15 to 90% and most preferably 15 to 85% by weight of water. Creams, lotions and gels may comprise lesser amount of water in the range of 5 to 95 % by weight, more preferably 10 to 90% by weight and most preferably 20 to 90% by weight of water. In some scenario, the composition may comprise 20 to 90% by weight, more preferably 30 to 80% by weight and most preferably 40 to 70% by weight of water.

[0108] The leave-on composition according to the present invention may be formulated as a sanitizer composition that is intended for providing antimicrobial effect against harmful or potentially harmful microorganisms that may be present on the skin. Sanitizers preferably comprise 40 to 98% by weight, preferably 45 to 95% by weight, preferably 50 to 90% by weight, preferably 55 to 85% by weight, preferably 60 to 80% by weight, preferably 65 to 75% by weight, e.g., 70% by weight of C2 to C4 monohydric alcohols wherein said monohydric alcohols are selected from ethanol, isopropyl alcohol, or mixtures thereof. Sanitizer may include certain polyols. Polyols in the composition according to the invention are preferably sorbitol, glycerol, polyethylene glycol, propylene glycol, and combinations thereof. Sanitizers may preferably comprise from 0.01 to 1% by weight of a thickening agent. More preferably the composition comprises 0.05 to 0.8% by weight of a thickening agent, most preferably 0.08 to 0.2% by weight of a thickening agent. Water is generally present in a sanitizer composition from 5 to 40% by weight, preferably 10 to 40% by weight, preferably 12 to 35% by weight, preferably 25 to 30% by weight. Sanitizer may preferably comprise one or more fragrance compounds.

[0109] Preferably, the composition comprises a range of other optional ingredients that include antioxidants, binders, buffering agents, colorants, astringents, fragrance, opacifying agents, conditioners, exfoliating agents, pH adjusters, skin sensitizers, skin soothing agents, sunscreens, and skin healing agents.P0000689 COM

[0110] 15

[0111] Wash-off compositions

[0112] The composition of the present invention may also be delivered through a wash-off composition. By ‘wash-off’, it is preferably meant that the composition applied on to a surface, e.g. skin, is removed from the surface preferably with water. In other words, the surface is rinsed-off or washed-off with water and the composition so applied is removed. Typically, if the composition is formulated in the form of a wash-off composition, then the composition so applied is removed preferably within 5 minutes, more preferably within 3 minutes, even more preferably within 1 minute and furthermore preferably within 30 seconds e.g. 10 or 15 seconds of it being applied on a surface.

[0113] The wash-off compositions of the invention preferably include a surfactant which may be one or more of anionic surfactant, cationic surfactant, non-ionic surfactant, or amphoteric surfactant.

[0114] In the wash-off composition the amount of surfactant varies based on the product format. For example, in a shampoo or wash composition, it preferably comprises the surfactant in an amount from 3 to 80% by weight, preferably 5 to 80% by weight surfactant. Further, if the wash-off composition is formulated as a solid wash-off composition, e.g. a bar, then the composition may comprise preferably from 25 to 70% by weight, preferably from 30 to 65% by weight, preferably from 35 to 60% by weight, preferably from 40 to 55% by weight of surfactant.

[0115] Alternatively, if the wash-off composition is formulated as a liquid wash-off composition, e.g. hand-wash or body-wash composition, then the composition preferably comprises from 3 to 20% by weight, preferably from 5 to 20% by weight, more preferably from 5 to 18% by weight, even more preferably from 5 to 15% by weight, further more preferably from 5 to 14% by weight and still more preferably from 5 to 12% by weight surfactant.

[0116] A suitable anionic surfactant may be selected from a soap and / or a non-soap synthetic anionic surfactant. Examples of soap include soaps that may be used as anionic surfactants in a wash-off composition include those formed after saponification of saturated fatty acids having carbon atoms in the range from C8 to C20 with an aqueous alkali e.g. sodium hydroxide, potassium hydroxide and ammonium hydroxide. Typical examples of soap include caprylate (C8), caprates (C10), laurate (C12), myristate (C14), palmitate (C16, stearate (C18) and oleate (C18:1) formed with an aqueous alkaliP0000689 COM

[0117] 16

[0118] mentioned earlier. Thus, preferred soaps are selected from potassium caprylate, potassium caprate, potassium laurate, potassium myristate, potassium palmitate, potassium stearate, potassium oleate, sodium caprylate, sodium caprate, sodium laurate, sodium myristate, sodium palmitate sodium stearate, sodium oleate and mixtures thereof. Often, soaps may be present as a mixture of soaps formed from fatty acids of varying chain lengths. Preferably, a non-soap synthetic anionic surfactants is selected from alkyl sulphates, alkyl ether sulphates (AES), alpha olefin sulfonates (AOS), isethionates, taurates and mixtures thereof.

[0119] When the composition is in liquid form, it is preferred that the anionic surfactant is a soap, an isethionate, taurate, glycinate, glutamate, alkyl sulphate, a-olefin sulphonate, alkyl ether sulphate, succinate, sulphosuccinate, sarcosinate, amphoacetate, and mixtures thereof.

[0120] Preferably, the anionic surfactant is selected from soap, alkyl sulphates, alkyl ether sulphates, isethionate, taurate, and mixtures thereof. Even more preferably, alkyl ether sulphates are used as the anionic surfactant in the composition. Anionic surfactants are known to provide foam and cleansing action.

[0121] Examples of alkyl sulphates that may be used as anionic surfactant in the composition include sodium lauryl sulphate (SLS), sodium myristyl sulphate, ammonium lauryl sulphate (ALS), sodium pareth sulphate, and mixtures thereof.

[0122] Examples of AES that may be used as non-soap anionic surfactants include anionic surfactants of the general formula:

[0123] R1-(OR’)n-O-SO3- M+,

[0124] wherein:

[0125] R1 is saturated or unsaturated C8-C16, preferably C12-C14 alkyl chain; preferably, R1 is a saturated C8-C16, more preferably a saturated C12-C14 alkyl chain;

[0126] R’ is ethylene; n is from 1 to 18; preferably from 1 to 15, more preferably from 1 to 10 and even more preferably from 1 to 5,

[0127] M+ is a suitable cation which provides charge neutrality, preferably sodium, calcium, potassium, or magnesium, ammonium, more preferably a sodium cation or ammonium.P0000689 COM

[0128] 17

[0129] Examples of AES that may be used as non-soap anionic surfactants in the composition include sodium lauryl ether sulphate (SLES), sodium myristyl ether sulphate and sodium palmityl ether sulphate, ammonium lauryl ether sulphate (ALES) and mixtures thereof. Preferred AES is SLES having 1 to 3 ethylene oxide units per molecule. SLES having 1 to 2 ethylene oxide units per molecule is more preferred.

[0130] Another class of anionic surfactants which is particularly desirable are alkyl isethionate (e.g., fatty acid esters of isethionate acid such as sodium lauroyl or sodium cocoyl isethionate) and alkyl taurate (e.g., alkyl taurate amides such as N-methyl taurate). Especially suitable is to include them as a surfactant mixture as they have the advantage that they are sulphate-free. Further, they offer the ability to formulate isotropic systems at neutral and slightly acidic pH.

[0131] In a preferred aspect, the present invention relates to a composition comprising 5 to 20% by weight, preferably 6 to 15% by weight of a surfactant system which comprises (a) alkali metal acyl isethionate, (b) alkali metal alkyl taurate. Preferred isethionate sulfonates include cocoyl isethionate and lauroyl isethionate, preferably having sodium or potassium as counterions. The other preferred anionic surfactant for inclusion in the composition of the invention are alkyl taurates, e.g. alkyl taurate amides. Preferably alkyl taurate amides include sodium methyl cocoyl taurate and sodium methyl lauroyl taurate.

[0132] The composition may also be delivered with soap as the anionic surfactant or as a combination of one or more of the above mentioned synthetic anionic surfactants together with soap. Such combination of soap and synthetic anionic surfactants may be included in a cleansing composition in both liquid as well as in solid (e.g. a bar) compositions. The soap for preparing the cleansing composition of the invention is preferably a C8-C24 soap, more preferably C10-C20 soap and most preferably C12-C18 soap. The cation of the soap can be alkali metal, alkaline earth metal or ammonium. Preferably, the cation of the soap is selected from sodium, potassium or ammonium. More preferably the cation of the soap is sodium or potassium. Fatty acids derived from other suitable oils / fats such as groundnut, soybean, tallow, palm, palm kernel, etc. may also be used in other desired proportions. Such combination of soap with synthetic surfactants is often called a soap-syndet formulation.P0000689 COM

[0133] 18

[0134] Preferably, the composition may further comprise an amphoteric surfactant. They provide foam boost and improve sensorial of the composition. Preferably, amphoteric surfactants are selected from the class of betaines, sultaine, ethanolamide. Preferred examples are cocamidopropyl betaine (CAPB), cocoamphoacetate and mixtures thereof. When included the composition preferably comprises from 0.1 to 40% by weight, more preferably 1 to 20% by weight, most preferably from 1 to 5 %by weight of the amphoteric surfactant.

[0135] Non-ionic surfactants may also be included in the wash-off compositions. Suitable nonionic surfactants are the same as those described under the leave-on composition section above. The composition may further comprise an amphoteric surfactant selected from betaines, e.g. cocamidopropyl betaine (CAPB), sulphobetaine, cocoamphoacetate, and mixtures thereof. Most preferred amphoteric surfactant is CAPB. Amphoteric surfactants provide foam boost and improve sensorial properties of the composition.

[0136] The pH of the composition is preferably in the range of 4.0 to 10.5, more preferably 4.0 to 9.0. pH as reported in the present invention is measured as follows: the composition is diluted 1:1 with deionized water and pH is directly measured by a standard pH meter at room temperature (~25°C).

[0137] The composition when in the liquid form preferably comprises 5 to 95% by weight, more preferably in 40 to 90% by weight of water. In a liquid wash-off format the composition may comprises water in an amount from 50 to 95% by weight, more preferably from 60 to 90% by weight, even more preferably from 70 to 85% by weight, furthermore preferably from 75 to 80% by weight and still more preferably from 80 to 85% by weight of water.

[0138] Preferably, the composition further comprises water soluble / dispersible polymers. They are known to increase the viscosity and stability of liquid cleansing compositions, to enhance in-use and after-use skin sensory feels, and to enhance lather creaminess and lather stability. Such polymers may preferably be used in amounts from 0.1 to 10% by weight, more preferably from 0.1 to 5% by weight and most preferably 0.1 to 3% by weight of the composition.P0000689 COM

[0139] 19

[0140] Examples of water soluble / or dispersible polymers include the carbohydrate gums such as cellulose gum, microcrystalline cellulose, cellulose gel, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethylcellulose, methyl cellulose, ethyl cellulose, guar gum, gum karaya, gum tragacanth, gum arabic, gum acacia, gum agar, xanthan gum and mixtures thereof; modified and nonmodified starch granules and pregelatinized cold water soluble starch; emulsion polymers such as Aculyn® 28, Aculyn® 22 or Carbopol® Aqua SF1; cationic polymer such as modified polysaccharides including cationic guar available from Rhone Poulenc under the trade name Jaguar® C13S, Jaguar® C14S, Jaguar® C17, or Jaguar® C16; cationic modified cellulose such as LICARE® Polymer JR 30 or JR 40 from Amerchol; N-Hance® 3000, N-Hance® 3196, N-Hance® GPX 215 or N-Hance® GPX 196 from Hercules; synthetic cationic polymer such as Merquat® 100, Merquat® 280, Merquat® 281 and Merquat® 550 sold by Nalco; cationic starches such as StaLok® 100, 200, 300 and 400 sold by Staley Inc.; cationic galactomannans such as Galactasol® 800 series by Henkel, Inc.; Quadrosoft® LM-200; and Polyquaternium-24®. Also suitable are high molecular weight polyethylene glycols such as Polyox® WSR-205 (PEG 14M), Polyox® WSR-N-60K (PEG 45), and Polyox® WSR-301 (PEG 90M).

[0141] The composition may further comprise emollients. Emollients listed above for use in leave-on compositions may be in wash-off compositions also.

[0142] The compositions may further comprise a wide range of other optional components e.g., antioxidants, biological additives, buffering agents, colorants, astringents, fragrance, humectants, opacifying agents, conditioners, pH adjusters, skin soothing agents, and skin healing agents.

[0143] The composition may also be delivered in solid form in which case it is preferably in the form of a bar. In such cases soap bars are preferred. Soap bars generally have the following constitution. Soap, i.e. salt of fatty acid is preferably present in an amount of 5 to 80 % by weight, preferably from 10 to 75% by weight, more preferably 25 to 75% by weight of the cleansing composition. It may comprise small amount, about 0.1 to 10% by weight, of synthetic anionic surfactants which could be one or more of the ones listed herein above. The soap bars may include other known ingredients such as perfumes, pigments, preservatives, emollients, gelling agents, and thickening agents. Choice of these ingredients will largely depend on the format of the composition. Water is aP0000689 COM

[0144] 20

[0145] preferred carrier. When water is present, it is preferably present in at least 1 % by weight, more preferably at least 2% by weight, furthermore preferably at least 5% by weight by weight of the composition. When water is the carrier, soap bar may comprise 10 to 50% by weight, more preferably 12 to 40% by weight, furthermore preferably from 12 to 35% by weight of water.

[0146] The cleansing composition of the invention may also be delivered through a moisturizing bar composition. Moisturizing bar compositions comprising fatty acyl isethionates, e.g. cocyl isethionate, are especially preferred. Fatty acyl isethionates surfactant "products" are defined as mixtures of anionic acyl isethionate surfactants and fatty acids / fatty acid soaps. They are highly desirable in skin or hair cleansing products because they lather well, are mild to the skin and have good emollient properties. Typically, fatty acid isethionate surfactant products are produced by esterification of fatty acids or by reaction of fatty acid chloride having carbon chain length of C8 to C20 with isethionate. A typical surfactant product containing fatty acyl isethionate contains about 40 to 95% by weight acid isethionate, and 5 to 50% by weight, typically 10 to 40% by weight of free fatty acid, may further contain less than 5% by weight, more preferably less than 2% by weight or trace level of other additives. Fatty acid soap may be included in the range of 5 to 15% by weight. Other surfactants like betaines may be included in 1 to 5% by weight. Water is generally included in 2 to 8% by weight of the composition.

[0147] The composition of the invention may also be delivered through a self-foaming composition. A preferred self-foaming composition comprises 2.5 to 10% by weight surfactant which is usually a mixture of anionic surfactant and amphoteric surfactant. Anionic surfactants preferred are of the amino acid class viz. sodium lauroyl glycinate, sodium cocoyl glycinate, sodium lauroyl glutamate, sodium cocoyl glutamate, and may also be selected from one or more of sodium lauroyl isethionate, sodium cocoyl isethionate, alpha olefin sulfonate (AOS), or a combination thereof. The amphoteric surfactant may be one or more as listed herein above. Self-foaming composition additionally usually comprises a humectant selected from glycerol, propylene glycol, dipropylene glycol, polypropylene glycol, polyethylene glycol, sorbitol, hydroxypropyl sorbitol, hexylene glycol, 1,3-butylene glycol, isoprene glycol, 1,2,6-hexanetriol, ethoxylated glycerol, propoxylated glycerol, or a combination thereof. It may also comprise an emollient which may be selected from one or more of the compounds listed herein above. Such types of self-foaming compositions are usually free of sulphate.P0000689 COM

[0148] 21

[0149] The composition preferably further comprises preservatives to protect against the growth of potentially harmful microorganisms. Examples of ingredients that may be used as preservatives in the composition include alkyl esters of para hydroxybenzoic acid, hydantoin derivatives, propionate salts, and a variety of quaternary ammonium compounds. More preferably, ingredients that may be used as preservative in the composition are sodium benzoate, iodopropynyl butyl carbamate, methylisothiazolinone, iodopropynylbutylcarbamate, phenoxyethanol, methyl paraben, propyl paraben, imidazolidinyl urea, sodium dehydroacetate, ethylhexylglycerin, benzyl alcohol, alkane diols, and mixtures thereof. The alkane diols that are suitable for use as preservative are C6-C12 alkanes that are vicinally substituted with hydroxy groups. Illustrative examples include 1,2-octane diol (caprylyl glycol), 2,3-octane diol, 1,2-nonane diol, 1,2-decane diol, 1,2-hexane diol, 3,4-octane diol, mixtures thereof or the like where caprylyl glycol is typically the most preferred. When present in the composition, preservatives are added preferably in an amount 0.001 to 5% by weight, more preferably 0.01 to 3% by weight and most preferably 0.02 to 2% by weight.

[0150] The composition as per the invention may also be delivered through a make-up remover composition. Make-up remover compositions in various forms are known and available like micellar water, lotions, foams, gels, and creams. A preferred make up remover is one which has a high amount of oils, preferably in the range of 80 to 90% by weight, surfactant, preferably non-ionic, in the range of 5 to 15% by weight, optionally emollients up to 10% by weight and the rest may be water.

[0151] The composition being wash-off or leave-on format may further comprise one or more skin brightening agents. Such one or more skin brightening agents may be selected from vitamin B6, vitamin C and its derivatives, vitamin E, glutathione precursors, resorcinol, phenyl ethyl resorcinol, 4-alkyl resorcinol e.g. 4-ethyl resorcinol, 4-butyl resorcinol, 4-hexyl resorcinol, and mixtures thereof. Alternatively, the one or more skin brightening agents may also be selected from galardin, adapalene, ammonium lactate, arbutin, butyl hydroxy anisole, butyl hydroxy toluene, citrate esters, deoxyarbutin, 1,3-diphenyl propane derivatives, 2,5-dihydroxybenzoic acid and its derivatives, 2-(4-acetoxyphenyl)-1,3-dithiane, 2-(4-hydroxyphenyl)-1,3-dithiane, ellagic acid, glucopyranosyl-1 -ascorbate, gluconic acid, glycolic acid, 4-Hydroxy-5-methyl-3[2H]-furanone, 4-hydroxyanisole and its derivatives, 4-hydroxybenzoic acid derivatives, hydroxycaprylic acid, inositol ascorbate, linoleic acid, magnesium ascorbyl phosphate,P0000689 COM

[0152] 22

[0153] 5-octanoyl salicylic acid, salicylic acid, 3,4,5-trihydroxybenzyl derivatives, acetylglucosamine, pitera extract, symwhite, calcium pantothenate (Melano-block), seppiwhite and mixtures thereof. Such one or more skin brightening agents may preferably be present in an amount from 0.1 to 10% by weight, more preferably from 1 to 7% by weight and even more preferably from 3 to 5% by weight.

[0154] The composition may comprise an organic sunscreen selected from one or both of a UVA sunscreen and a UVB sunscreen. Sunscreens include those materials commonly employed to block ultraviolet light. Illustrative compounds are the derivatives of PABA, cinnamate and salicylate. For example, avobenzophenone (Parsol 1789®) octyl methoxycinnamate and 2-hydroxy-4-methoxy benzophenone (also known as oxybenzone) can be used. Octyl methoxycinnamate and 2-hydroxy-4-methoxy benzophenone are commercially available under the trademarks, Parsol MCX and Benzophenone-3, respectively. The exact amount of sunscreen employed in the compositions can vary depending upon the degree of protection desired from the sun’s UV radiation. Additives that reflect or scatter the sun rays may also be employed. These additives include oxides like zinc oxide and titanium dioxide.

[0155] There is provided a method for enhancing antimicrobial peptides (AMP) secretion on an external surface of the human body comprising steps of (a) applying the composition according to the present invention topically on to the surface, and (b) optionally, rinsing the surface. The method is non-therapeutic or cosmetic in nature.

[0156] As specified earlier in this specification, antimicrobial peptides (AMP) are typically integral part of the skin’s own defence mechanism. It is reported that AMPs are ubiquitous in nature, and they typically exhibit a broad spectrum of activity against harmful or non-commensal microorganisms. It is reported that enhancing AMP secretion and / or its functionality helps in delivering antimicrobial benefit in an ‘eco-friendly’ manner, i.e. it provides protection form harmful or non-commensal microorganisms. Preferably, harmful or non-commensal microorganisms are one or more of Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), Pseudomonas aeruginosa (P. aeruginosa), Staphylococcus hominis (S. hominis), and Cutibacterium acnes (C. acnes). The term ‘protecting’ herein implies of inhibition or removal of non-commensal microorganisms. Without wishing to be bound by theory, it is submitted that the inhibition or removal of non-commensal microorganism is obtained by improving secretion or boostingP0000689 COM

[0157] 23

[0158] antimicrobial peptides present in the external surface of the human body, mostly on skin. Said benefit is obtained by non-therapeutically and obtained by cosmetically applying the composition. It is submitted that removal or inhibition of non-commensal microorganisms is not necessarily prophylactic. For example, it is well-known that stimulating a healthy microbiome balance, by suppressing the growth of S. aureus aids in maintaining the barrier function and thereby the improving moisture retention of the skin. It is submitted that somewhat reduced barrier function clearly is an inconvenience but does not represent a pathological state. Therefore, this evidently represents a nonprophylactic realisation of the claimed use / method.

[0159] The composition can be in the form of a liquid, lotion, cream, foam, scrub, gel, soap bar or toner, or applied with an implement or via a face mask, pad or patch. Non-limiting examples of such compositions include leave-on skin lotions and creams, shampoos, conditioners, shower gels, toilet bars, antiperspirants, deodorants, depilatories, lipsticks, foundations, mascara, sunless tanners, and sunscreen lotions.

[0160] Step (a) may be carried out simply by taking a suitable amount, for example 5 mL or 2 grams, depending on whether the composition is in liquid or solid form, of the composition on hands and gently spreading it over an external surface, e.g. skin. The composition, if required, may also be applied using an applicator in which case, a user is expected to use the applicator to carry out the step of applying the composition; and spread the composition using the applicator on the surface.

[0161] If step (b) is carried out, it is preferably be carried out within 5 minutes, more preferably within 4 minutes, even more preferably within 3 minutes, furthermore preferably within 2 minutes and yet more preferably within 1 minute, e.g. 30 seconds, or 15 seconds, from carrying out step (a). Such optional step (b) is particularly required to be carried out within 5 minutes, if the composition is formulated into a wash-off type of composition, e.g. a face wash composition. The method is non-therapeutic / cosmetic in nature.

[0162] In another aspect there is provided a use of combinations of one or more compounds of formula I, II or III and the antimicrobial peptides (AMP) and / or the AMP boosters for enhancing secretion of antimicrobial peptides when applied on external surfaces of the human body, e.g. skin. Preferably the use is non-therapeutic or cosmetic in nature. Preferably the compounds of formula I, II or III in the use is selected from levulinic acid,P0000689 COM

[0163] 24

[0164] methyl levulinate, ethyl levulinate, propyl levulinate, butyl levulinate, and benzyl levulinate. . It is submitted that enhancing secretion of AMPs may not be necessarily prophylactic. For example, it is well-known that AMPs are part of skin own defence system, and it may stimulate a healthy microbiome balance, by suppressing the growth of non-commensal bacteria, like, S. aureus. Such supression aids in maintaining the barrier function and thereby the improving moisture retention of the skin. It is submitted that somewhat reduced barrier function clearly is an inconvenience, but does not represent a pathological state. Therefore, this evidently represents a non-prophylactic realisation of the claimed use.

[0165] The invention will now be illustrated with the help of the following non-limiting examples:

[0166] Examples

[0167] Effect of the compounds of formula I, II or III on gene expression levels of an AMP or AMP protein secretion was performed on primary keratinocyte cells and in 3D Living Skin Equivalent (LSE). The following protocol was used for AMP gene expression or AMP protein secretion in primary cells.

[0168] Materials:

[0169] Normal Human Epidermal Keratinocyte cells (NHEK), EpiLife Keratinocyte Growth Medium (KGM) (with antibiotics Pen-Strep), Dulbecco’s Phosphate Buffered Saline (DPBS), Trypsin-EDTA, Trypsin Neutralizer Solution, Ethanol, RNA extraction kit (RNeasy kit), RLT buffer, RT-PCR kit.

[0170] Procedure:

[0171] i. Thaw the NHEK cells and expand them in KGM in 2 -25 cc and then 75 cc flasks to 80% confluency, changing media every alternate day and incubating at 37°C in a 5% CO2 incubator.

[0172] ii. Discard spent media and rinse the adhered cells in the flask with DPBS. Add Trypsin- EDTA and tap to dislodge the adhered cells. Add Trypsin Neutralizer and gently mix it.

[0173] iii. Transfer the contents to a 15 -ml tube and spin down the cells. Discard supernatant and resuspend cells in KGM. Count and seed 30,000-50,000 cells per well in 24 well TC plates, incubate to 80-85% confluence (~2-3 days). Change media every alternate day.P0000689 COM

[0174] 25

[0175] iv. Add differentiation media (KGM with 2 mM CaCI2) and incubate for 2 days.

[0176] v. Prepare actives in KGM at the required concentration (final concentration of ethanol solvent, if used, should be less than 0.1%).

[0177] vi. Replace the differentiation media with the actives containing media and incubate for 24 h (for gene expression) or 72 h (for AMP protein secretion).

[0178] vii. For gene expression part, after 24 hours exposure to the active, discard the supernatant and wash cells once with DPBS. Add 250 pL of RLT buffer to each well to lyse the cells and transfer the entire content to 1.5 mL tubes. Store at -80°C until extraction.

[0179] viii. Perform RNA extraction as per the kit, quantify the RNA, do cDNA synthesis and do real time qPCR forcDNA samples. Calculate the relative expression level over control normalised to housekeeping gene.

[0180] ix. For AMP protein measurement, aliquots of cell-free spent medium are withdrawn from the wells at 72 h post exposure to actives, added to tubes and stored at -80°C until AMP protein measurement.

[0181] x. Specific AMP (Psoriasin) was measured in cell-free spent medium using a commercially available ELISA kit as per manufacturer’s recommendation.

[0182] The following protocol was used to test the actives on 3D Living Skin Equivalent (from Mattek). Twelve-well plates indicating the treatment / exposure conditions were labelled and 1 mL of medium was added in to each well. Tissue from the agarose-containing package was removed carefully using sterile forceps and transferred into the labelled 12 well plates containing media, and the plates were incubated at 37°C, 5% CO2 incubator for 12 to 18 hours. After the overnight incubation, under sterile conditions the lid was removed from the 12-well plate and 1 mL / well of fresh, pre-warmed medium was added in to each well of the plates. The tissues were then carefully blotted and placed on a sterile plate for dosing. The actives were prepared at the required concentration as a solution in water and sonicated using a bath sonnicator for 10 minutes. Each tissue was dosed with 10 pL of the prepared solution and the inserts were then placed in the well. A vehicle control (sonicated water) was also included in experiment. One set of 3 tissues was retained as a no treatment control. Each active was tested in three replicates. The plate was then incubated at 37°C, 5% CO2 incubator for 24 hrs. On day 2, the plates were removed from the incubator; and all the tissues were rinsed with sterile Dulbecco’s phosphate buffered saline. After that, all tissues were cut using a sterile scalpel and then immersed in 500 pL of RNeasy lysis buffer with p-Mercaptoethanol (100:1) in a labelledP0000689 COM

[0183] 26

[0184] 1.5 mL Eppendorf tube. RNA from these tissues was extracted and probed by qPCR for amplification of AMP genes and the fold change over the vehicle control was calculated.

[0185] Enhanced AMP gene expression obtained by gamma carboxylic acid / esters treatments of primary keratinocytes are shown below:

[0186] Table 1

[0187]

[0188] P0000689 COM

[0189] 27

[0190]

[0191] Enhanced psoriasin protein secretion by gamma carboxylic acids / esters treatments of primary keratinocytes are shown below:

[0192] Table 2

[0193]

[0194] P0000689 COM

[0195] 28

[0196]

[0197] Enhanced AMP gene expression obtained by gamma carboxylic acids / esters treatments on 3D Living Skin Equivalent are shown below:

[0198] Table 3

[0199]

[0200] Measurement of Antimicrobial efficacy:

[0201] Culture preparation

[0202] E. coli or S. aureus was inoculated from glycerol stock to TSA slants. Slants were incubated at 37°C aerobically for 24 hours until saturated growth is seen. These slants were then used to streak TSA agar plates which were then incubated at 37°C for about 16 hrs. One loopful of this plate culture was suspended (pipetting and vortex mixing) inP0000689 COM

[0203] 29

[0204] 10 mM sodium phosphate buffer pH 5.8 (~10ml of buffer). Optical density (OD) was checked, and culture was adjusted to 0.6 OD for E. coli and 0.2 for S. aureus at 620 nanometres. These OD values correspond to about a cell number of 1-5 x 108cfu / mL for the respective microorganism. The culture was serially diluted 10-fold to get 1-5 x 106cfu / mL.

[0205] AMP

[0206] Working stocks of 10 pg / mL of LL37 (an AMP) was prepared by diluting it in water. This working stock was used for adding to the reaction wells as shown in tables below where 0.1 pg / mL LL37 is equal to 0.00001% by weight.

[0207] The test reaction was prepared in a 96 well microtiter plate with duplicate wells for each reaction. From the working stock of LL37 15 pL and 22.5 pL were added to the respective wells for 0.5 pg / mL and 0.75 pg / mL test concentrations. Fatty acid working stocks of 100 times higher the tested concentrations were prepared in DMSO and 3 pL of this was added in the respective wells. The total volume of the reaction in each well was made to 300 pl with water. The Contact time provided was 4 hours at 37°C. Postincubation, the reactions were plated on pre-prepared TSA agar plates in requisite dilutions. Dilutions were made in D / E neutralizer, in a microtitre plate (270 pl of D / E + 30 pl reaction mixture, etc.). 100 pl volume of desired dilutions is spread plated onto prepared plates. Plates were incubated aerobically overnight. Colonies were counted at the end of the incubation to quantitate the log kill / survival. A difference of 0.5 and above of the combination tested for synergy over the sum of the individual activity / kill is considered a synergy.

[0208] Antimicrobial efficacy obtained by combination of gamma carboxylic acid and antimicrobial peptide (AMP) is shown below:P0000689 COM

[0209] 30

[0210] Table 4

[0211]

[0212] Antimicrobial efficacy against target microbes obtained by combinations of esters of gamma carboxylic acids and antimicrobial peptide (AMP):

[0213] Table 5

[0214]

[0215] Further, Example 29 in the form of a bodywash composition was prepared following the recipe provided in table 6.P0000689 COM

[0216] 31

[0217] Table 6

[0218]

[0219] It is understood that the experiments described above were conducted in an in vitro assay to evaluate the synergistic antimicrobial behaviour. It is expected that the concentrations to be actually used to prepare a composition for topical use would be vastly different. The concentrations could be orders of magnitude higher due to the following reasons that affect the difference in concentration in the bulk as compared to that at the cellular level. The composition may be formulated as an emulsion or a gel with additional ingredients. The concentration of the desired actives in the oil phase and in the water phase, is expected to be different. They may also have very different physical and hydrodynamic properties like partition coefficients, diffusional rates, convective transport rates and rheological properties. Therefore, it is expected that the concentrations to be used when formulated as a composition would be different from that at the cellular level.

Claims

P0000689 COM32Claims:

1. A skin care composition comprising:a. 0.000001 to 10% by weight of one or more compounds of formula I or II or III:where:R1 is selected from H, a straight or branched C1 to C6 alkyl group or C2 to C6 allyl group, an aryl group, Na+, K+, Ca2+, Mg2+, NH4+;R2 is selected from H, a straight or branched C1 to C14 alkyl group, an ethylamine group (CH2NH2) for formulae I or II;R2 is H or a straight or branched C1 to C4 alkyl group for formula III; and R3 is selected from Cl Br, I’ and NH2; andb. an antimicrobial peptide (AMP) selected from defensins, histatins, cathelicidin, dermcidin, S100 peptides, RNase peptides, and combinations thereof; and / or an antimicrobial peptide booster selected from hydroxystearic acid, nicotinamide, isomer of nicotinamide, cycloalkyl derivatives of nicotinamide, Vitamin A, Vitamin C, and linseed oil and combinations thereof,wherein the composition is free of cantharidin.

2. The composition comprising:a. one or more compounds of formula I or II as claimed in claim 1 ; and b. an antimicrobial peptide and / or a booster thereof.P0000689 COM333. The composition as claimed in claims 1 or 2 wherein the composition comprises one or more compounds of formula I or II where R2 is selected from H, a straight or branched C1 to C10 alkyl group.

4. The composition as claimed in any one of claims 1 to 3 wherein the composition comprises one or more compounds of formula I or II where R2 is selected from H, a straight or branched C1 to C5 alkyl group.

5. The composition as claimed in any one of claims 1 to 4 wherein the composition comprises a compound of formula I where:- R1 is H; and R2 is C1, which corresponds to levulinic acid; or- R1 is Na+or K+or Ca2+or Mg2+or NH4+; and R2 is C1, which corresponds to a salt of levulinic acid; or- R1 is a straight or branched C1 to C6 alkyl group or an C2 to C6 allyl group; or an aryl group; and R2 is C1 which corresponds to an ester of levulinic acid.

6. The composition as claimed in any one of claims 1 to 4 wherein the composition comprises a compound of formula I where:- R1 is H; and R2 is an ethylamine group (CH2NH2) which corresponds to amino levulinic acid; or- R1 is Na+or K+or Ca2+or Mg2+or NH4+; R2 is an ethylamine group (CH2NH2), which is a salt of amino levulinic acid; or- R1 is a straight or branched C1 to C6 alkyl group or a C2 to C6 allyl group; or an aryl group; and R2 is an ethylamine group (CH2NH2) which corresponds to an ester of amino levulinic acid.

7. The composition as claimed in any one of claims 1 to 6 wherein the antimicrobial peptide is selected from defensins, histatins, S100 peptides, RNase peptides, and combinations thereof.

8. The composition as claimed in any one of claims 1 to 7 wherein the booster of antimicrobial peptides is selected from hydroxy-stearic acid, nicotinamide, linseed oil, and combinations thereof.P0000689 COM349. The composition as claimed in any one of claims 1 to 8 wherein the pH of the composition is in the range from 4 to 11.

10. The composition as claimed in any one of claims 1 to 9 wherein the composition is a leave-on composition.

11. The composition according to any one of claims 1 to 9 wherein the composition is a wash-off composition.

12. A non-therapeutic method for enhancing secretion of antimicrobial peptides (AMP) on an external surface of the human body comprising steps of:a. applying topically a composition as claimed in any one of claims 1 to 11 on to the surface; andb. optionally, rinsing the surface.

13. Use of a combination of:a. one or more of compounds of formula I or II or III:where:R1 is selected from H, a straight or branched C1 to C6 alkyl group or C2 to C6 allyl group, an aryl group, Na+, K+, Ca2+, Mg2+, NH4+;R2 is selected from H, a straight or branched C1 to C14 alkyl group, an ethylamine group (CH2NH2) for formulae I or II;R2 is H or a straight or branched C1 to C4 alkyl group for formula III; and R3 is selected from Cl Br, I’ and NH2; andP0000689 COM35b. an antimicrobial peptide booster selected from hydroxy-stearic acid, nicotinamide, isomer of nicotinamide, cycloalkyl derivatives of nicotinamide, Vitamin A, Vitamin C and linseed oil, and combinations thereoffor enhancing secretion of antimicrobial peptides (AMP) when applied on an external surface of the human body.

14. Use as claimed in claim 13 wherein the one or more compounds of formula I is / are selected from levulinic acid, methyl levulinate, ethyl levulinate, propyl levulinate, butyl levulinate and benzyl levulinate.

15. Composition as claimed in any one of claims 1 to 11 for use in enhancing secretion of antimicrobial peptides (AMP) when applied on an external surface of the human body thereby protecting from harmful or non-commensal microorganisms.

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