COMPOSITIONS CONTAINING A PEPTIDE CELL GROWTH PROMOTER

Abstract: The invention concerns compositions for topical application to skin and hair comprising glutamine-rich peptides, which have a molecular weight of 150-1000 D, to enhance cell growth, particularly in the hair follicle. The peptides are preferably obtained by hydrolysis of vegetable proteins such as wheat gluten and preferably have a glutamine content of at least 5 %.

Description: COMPOSITIONS CONTAINING A PEPTIDE CELL GROWTH PROMOTOR FIELD OF THE INVENTION The invention relates to cosmetic and pharmaceutical compositions for topical application to mammalian skin or hair, containing a peptide cell growth promoter which is capable of increasing or maintaining cell growth. The invention particularly relates to such compositions intended for application to the human scalp, to promote hair growth thereon. BACKGROUND In most mammals hair does not grow continuously, but undergoes a cycle of activity involving alternate periods of growth and rest. The hair growth cycle can be divided into three main stages, i.e. : i. the growth phase known as anagen, during which the hair follicle penetrates deeply into the dermis with the cells of the bulb dividing rapidly and differentiating to form the hair, ii. the transitional stage known as catagen, which is heralded by the cessation of mitosis, and during which the follicle regresses upwards through the dermis and hair growth ceases, iii. the resting stage known as telogen, in which the regressed follicle contains a small secondary germ with an underlying ball of tightly packed dermal papilla cells. The initiation of a new anagen phase is revealed by rapid proliferation in the germ, expansion of the dermal papilla and elaboration of basement membrane components. The hair cycle is then repeated many times until, as a consequence of the onset of male pattern baldness, most of the hair follicles spend an increasing proportion of their time in the telogen stage, and the hairs produced become finer, shorter and less visible; this is known as terminal to vellus transformation. PRIOR ART Although there have been many claims in the scientific literature to the promotion or maintenance of hair growth by the topical application of hair tonics and the like, few have been shown to be sufficiently free from disadvantageous clinical side effects, whether administered topically, orally or systemically, to warrant commercial exploitation as an ethical pharmaceutical, proprietary medicine, or as a cosmetic product. Among the many hair regrowth studies that have been reported in the literature, there is included the work of Bazzano as described in WO 85104577. This publication describes a composition which is useful for increasing the rates of hair growth on mammalian skin, prolonging the anagen phase of the hair growth cycle and for treating various types of alopecias. The composition in question comprises a pyrimidine carbamate. It has also been reported in US 4,139,619 to Chidsey assigned to the Upjohn Company, that a topical composition comprising minoxidil as the free base or acid addition salt thereof, or certain specif ied related iminopyrimidines, is useful in stimulating the conversion of vellus hair to growth as terminal hair, as well as increasing the rate of growth of terminal hair. In spite of the apparent stimulation of hair growth or regrowth reported independently by Bazzano and Chidsey, following topical application of minoxidil or related compounds, there is general concern that systemic side- effects can result, particularly following topical application of minoxidil. Thus it is generally recognised in the medical literature that the side effects of orally administered minoxidil are very serious and include fluid retention, tachycardia, dyspnea, gynecomastia, fatigue, nausea and cardiotoxicity. There is also evidence that certain side effects have been experienced following topical application of minoxidil . In EP 451 903 (The Proctor & Gamble Company) and literature cited therein it is reported that various products derived from dermal papilla cells, including certain polypeptides with specific isoelectric pH and molecular weights (between 25 and 45 kD) , would be able to stimulate hair growth. EP 225 639 (Meiji Seika Kaisha Ltd) describes the stimulation of hair growth by local application of the peptide hormone VIP, which is a basic polypeptide having a molecular weight of 3326. In JP 61151109 (Lion Corp.) it is reported that compositions comprising a mono-N-long chain acyl basic amino acid lower alkyl ester salt together with a higher fatty acid or aliphatic alcohol having an odd number of C atoms, or derivative thereof, can be used to regenerate or increase growth of hair. DE 1617477 (Fischer) discloses a hair tonic which is said to guarantee hair growth and includes a variety of amino acids and four vitamins. In EP's 490 581, 571 198 and 572 167 (all of Unilever) compositions comprising glutamine or derivatives or salts thereof or comprising glutamic acid derivatives or salts thereof are disclosed which induce, maintain or increase hair growth. An in vitro hair follicle growth test is described and the compounds are reported to stimulate follicle growth and hair production in this test. The glutamine derivatives include di- and small polypeptide derivatives in which the glutamine moiety is at the acidic end of the peptide chain. Especially in EP 490 581 and 571 198 glutamine is described as a potent promotor of hair growth which, however, has the disadvantage that it is not stable in aqueous skin and hair treatment compositions. Therefore, various derivatives have been disclosed in which the α-amino group is protected in order to stabilize the molecule. However, such molecules have to be prepared synthetically from glutamine. Thus, there is still a need for easily obtainable sources of glutamine, which are preferably of natural origin and are easily taken up by the cells of the skin, particularly the hair follicle. On the other hand, the use of polypeptides/protein hydrolysates in hair care products is known to improve various hair characteristics such as gloss and manageability. Thus, in DE 4309567 (Henkel KGaA) detergent mixtures for hair conditioning are disclosed which comprise protein hydrolysates obtained from hydrolysis of plant protein and having an average molecular weight of between 1000-10000 Dalton, preferably 2000-6000. The protein hydrolysates act as film forming agents. In JP 61 183298 it is disclosed to use specially purified polypeptides of average molecular weight between 300-2000 Dalton, obtained by hydrolysis of animal protein, such as collagen, keratin or silk, in cosmetic compositions, especially hair care products. They are said to be absorbed on hair. In DE 4324962 (Goldwell AG) hair care agents are disclosed which contain a protein hydrolysate and are said to improve shine, combability and vigour. Likewise, in SU 1796181 shampoo and shower gel preparations are disclosed containing oligopeptide with molecular weight of 700-2000, which give reduced electrostatic charging, more easy styling and a shiny appearance. Furthermore, DE 4207046 (=EP 562263, Goldwell) discloses a shampoo particularly for damaged hair, which comprises a plant protein hydrolysate containing hydroxyproline groups. JP 05 117129 discloses a dermatological drug containing a plant protein hydrolysate. It is intended for use in cosmetics, shampoo and soap to keep skin moisture and hair gloss. Likewise SU 1600774 discloses a shampoo containing hops, maltshoots extract and protein hydrolysate to improve structurising properties. In DD 243715 a protein hydrolysate for cosmetic application is disclosed, which is prepared from collagen or gelatin using a bacterial alkaline serine protease. JP 51 082737 discloses shampoo comprising a sucrose fatty acid ester, a protein, a peptide and an acylated peptid. The peptide is a vegetable or animal protein hydrolysate with a mol weight of < 10,000. the shampoo is said to have mildness, touch, foamability and foam stability. None of these products containing peptides or protein hydrolysates has in any way been connected to cell growth in the skin or the hair follicle or to hair growth. THE INVENTION It has now been found that small peptides in which glutamine is not necessarily at the acidic end of the chain can function as efficient glutamine sources for living cells. They are stable and easily taken up by the cell. Thus, they can be used to induce, increase and promote cell growth in mammalian skin, including the hair follicle. Accordingly the invention provides a composition, suitable for topical application to mammalian skin or hair, which comprises: i. a glutamine containing peptide or peptide mixture in an effective amount to induce, increase and promote cell growth in mammalian skin, with the proviso that if the peptide or peptides are prepared synthetically glutamine is not at the acidic end of the peptide chain. ii. a cosmetically acceptable vehicle. Preferably the peptides or peptide mixtures should have a molecular weight between 150 and 1000 D. Preferably the peptides or peptide mixtures are obtained by hydrolysis of proteins and should have a total glutamine content of 5% by weight or more, more preferably 10% or more. Such protein hydrolysates are preferably prepared by enzymatic hydrolysis. Protein hydrolysates which do not have the required glutamine content may be enriched in glutamine-containing peptides e.g. by suitable chromatographic techniques, or alternatively glutamine-rich hydrolysates may be obtained by the use of specific proteases, followed by separation of the glutamine-rich fraction. Preferred protein starting materials having a sufficiently high glutamine content are vegetable proteins such as wheat gluten and soy protein. The protein hydrolysates are preferable substantially free (less than 2% by weight, preferably less than 1%) of peptides above 10,000 Dalton and also preferably have a low content of free amino acids (less than 10% by weight, preferably less than 5%) . Peptides suitable for the invention as referred to above will hereinafter be referred to as "hair growth promotor" . The composition can comprise two more hair growth promoters, as herein defined. The total amount of the hair growth promoter present in the composition according to the invention is sufficient to induce, maintain or increase hair growth. This amount will depend on the effectiveness of the promoter, some being more effective than others, but in general an amount of from 0.001 to 99%, usually from 0.01 to 20% by weight of the composition will provide an adequate concentration for application to the skin, particularly the scalp, which can then be repeated as necessary to promote hair growth. The composition according to the invention also comprises a solid, semi-solid or liquid cosmetically and/or physiologically acceptable vehicle, to enable the hair growth promoter to be conveyed to the skin at an appropriate dilution. The nature of the vehicle will depend upon the method chosen for topical administration of the composition. The vehicle can itself be inert or it can possess physiological or pharmaceutical benefits of its own. The selection of a vehicle for this purpose presents a wide range of possibilities depending on the required product form of the composition. Suitable vehicles can be classified as described hereinafter. It should be explained that vehicles are substances which can act as diluents, dispersants, or solvents for the hair growth promoter which therefore ensure that they can be applied to and distributed evenly over the hair and/or scalp at an appropriate concentration. The vehicle is preferably one which can aid penetration of the hair growth promotor into the skin to reach the immediate environment of the hair follicle. Compositions according to this invention can include water as a vehicle, but they include at least one cosmetically acceptable vehicle other than water. Vehicles other than water that can be used in compositions according to the invention can include solids or liquids such as emollients, solvents, humectants, thickeners and powders. Examples of each of these types of vehicles, which can be used singly or as mixtures of one or more vehicles, are as follows: Emollients, such as stearyl alcohol, glyceryl monoricinoleate glyceryl monostearate, propane-1,2-diol, butane-1,3-diol, mink oil, cetyl alcohol, ispropyl isostearate, stearic acid, isobutyl palmitate, isocetyl stearate, oleyl alcohol, isopropyl laurate, hexyl laurate, decyl oleate, octadecan-2-ol, isocetyl alcohol, cetyl palmitate, dimethylpolysiloxane, di-n-butyl sebacate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, butyl stearate, polythylene glycol, triethylene glycol, lanolin, sesame oil, coconut oil, arachis oil, castor oil, acetylated lanolin alcohols, petroleum, mineral oil, butyl myristate, isostearic acid, palmitic acid, isopropyl linoleate, lauryl lactate, myristyl lactate, decyl oleate, myristyl myristate; Propellants. such as propane, butane, isobutane, dimethyl ether, carbon dioxide, nitrous oxide; Solvents, such as ethyl alcohol, methylene chloride, isopropanol, castor oil, ethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol monoethyl ether, dimethyl sulphoxide, dimethyl formamide, tetrahydrofuran; Humectants. such as glycerin, sorbitol, sodium 2-pyrrolidone5-carboxylate, soluble collagen, dibutyl phthalate, gelatin; Powders. such as chalk, talc, fullers earth, kaolin, starch, gums, colloidal silicon dioxide, sodium poly-acrylate, tetra-alkyl and/or trialkyl aryl ammonium smectites, chemically modified magnesium aluminium silicate, organically modified montmorillonite clay, hydrated aluminium silicate, fumed silica, carboxyvinyl polymer, sodium carboxymethyl cellulose, ethylene glycol monostearate. The amount of vehicle in the composition, including water if present, should preferably be sufficient to carry at least a portion of a selected hair growth promoter to the skin in an amount which is sufficient effectively to enhance hair growth. The amount of the vehicle can comprise the balance of the composition, particularly where little or no other ingredients are present in the composition. Accordingly, the vehicle or vehicles can comprise from 1 to 99.99%, preferably from 50 to 99.5% and ideally from 90 to 99% by weight of the composition. The composition according to the invention can also optionally comprise an activity enhancer. The activity enhancer can be chosen from a wide variety of molecules which can function in different ways to enhance the hair growth effects of the hair growth promoter. Some activity enhancers can also function as vehicles for the hair growth promotor. The amount of the activity enhancers, when employed in accordance with the invention, will normally be from 0.1 to 50% by weight of the composition, preferably from 0.5-25% and most preferably from 0.5 to 10%. Particular classes of activity enhancers include: (a) other hair growth stimulants, (b) penetration enhancers and (c) cationic polymers, whose presence can further improve the delivery of the ester through the stratum corneum to its site of action in the immediate environment of the hair follicle. (a) other hair growth stimulants are mentioned in EP 572167 (Unilever) and include substances which themselves possess the ability to stimulate or increase hair growth such as: i. Benzalkonium chloride, Benzethonium chloride, Phenol, Estradiol, Diphenhydramine hydrochloride, Chlorpheniramine maleate, Chlorophyllin derivatives, Cholesterol, Salicylic acid, Cystine, Methionine, Red pepper tincture, Benzyl nicotinate, dl-Menthol, Peppermint oil, Calcium pantothenate, Panthenol, Castor oil, Hinokitiol, Prednisolone, Resorcinol; ii. α-1,4 esterfied disaccharides described by Choay in EP 064012 and esterified oligosaccharides including at least one esterified disaccharide unit consisting of a uronic acid residue, as described in EP 211610 (Unilever) ; iii. Minoxidil glucuronides as described in EP 242967 (Unilever) ; iv. Minoxidil sulphates as described in WO 86/04231 (The Upjohn Co) ; v. Minoxidil and other derivatives thereof as described in US 4,139,619 (The Upjohn Co); vi. Ethylenediaminetetraacetic acid or salts thereof as described in US 4,814,351 (Redken Laboratories, Inc.) ; vii. Direct proteoglycanase inhibitors such as 1,10-phenanthroline, as described in EP 277428 (Unilever) ; viii.Glycosamineglycanase inhibitors as described in EP 277428 (Unilever) , such as aldonolactones, esterified aldonolactones, monosaccharides and esterified monosaccharides; ix Glycosaminoglycan chain cellular uptake inhibitors as described in EP 277428 (Unilever) , such as hexuronic acid and esters thereof; x. Chemical inhibitors of glycosidase activity, as described in EP 334586 (Unilever) , such as various lactams; xi. Chemical activators of protein kinase C enzymes as described in EP 334586, such as various diacylglycerols; xii. Glycosamineglycanase inhibitors as described in EP 348184, such as aldonomonolactone or alduronomonolactone derivatives; xiii.Glycosaminoglycanase inhibitors as described in EP 348184 (Unilever) such as acylated monosaccharides; xiv. Esters of pyroglutamic acid as described in US 4,774,255 (Lever Brothers Company) ; xv. Hexosaccharic acids or acylated hexosaccharic acids or salts or esters thereof, as described in EP 378388 (Unilever) ; xvi. Aryl-substituted ethylenes as described in EP 403238 (Unilever) ; xvii.N-acylated amino acids as described in EP 415598 (Unilever) . (b) Penetration enhancers, as has been stated earlier, can potentiate the benefit of the hair growth promoter by improving its delivery through the stratum corneum to its site of action in the immediate environment of the hair follicle close to the dermal papilla. The penetration enhancer can accordingly function in a variety of ways. It can for example, improve the distribution of the hair growth promoter on the skin surface or, it can increase its partition into the skin from the composition when applied topically, so aiding its passage to its site of action. Other mechanisms enhancing the benefit of the hair growth promotor may also be involved. Examples of penetration enhancers are mentioned in EP 572167. (c) Cationic polymers can potentiate the benefit of the hair growth promoter by improving its delivery to the hair and scalp. Examples of preferred cationic polymers include: Guar Hydroxypropyltrimonium chloride Quaternium-19 Quaternium-23 Quaternium-40 Quaternium-5? Poly(dipropyldiallylammonium chloride) Poly(methyl-γ-propaniodiallylammonium chloride) Poly(diallyl piperidinium chloride) Poly(vinyl pyridinium chloride) Quaternised poly (vinyl alcohol) Quaternised poly- (dimethylaminoethylmethacrylate) ; and mixtures thereof. Other adjuncts The composition according to the invention can also contain adjuncts other than those already mentioned, depending on the form of the intended product. It is, for example, possible to include antiseptics, preservatives, antioxidants, emulsifiers and colouring agents, pearlescers, foarn boosters, conditioning agents (such as cationic surfactants, cationic polymers and silicones) and agents such as PFPE (perfluoropolyethylene) for improving hair gloss, which can improve the stability and consumer appeal of the composition. The composition according to the invention can also be employed as a vehicle for a wide variety of cosmetically or pharmaceutically active ingredients, particularly ingredients which have some beneficial effect other than the promotion of hair growth when applied to the skin. The composition according to the invention may also comprise a perfume in an amount sufficient to make the composition acceptable to the consumer and pleasant to use. Usually, the perfume will form from 0.01 to 10% by weight of the composition. Surfactants The composition for use in the method according to the invention can be formulated as a shampoo and will then accordingly comprise one or more surfactants which are cosmetically acceptable and suitable for topical application to the hair. The amount of surfactant that can be present in the composition accordingly to the invention is up to 30%, preferably from 1 to 20% by weight of the composition. Examples of suitable shampoo surfactants in different classes are given below: Anionic surfactants, which are preferably chosen from alkyl sulphate, alkyl ether sulphate, alkyl sulphonate, alkyl aryl sulphonate, olefin sulphonate, acyl sarcosinate, acyl tauride, acyl isethionate, nonoalkyl sulphosuccinate, dialkyl-sulphosuccinate, acryl lactylate, acylated α-amino acid, alkyl carboxylate, monoalkyi phosphate and dialkyl phosphate. Specific examples of such anionic surfactants include: alkyl sulphates, such as sodium lauryl sulphate (eg. EMPICOL CX available from Albright & Wilson) , and triethanolamide lauryl sulphate (eg. EMPICOL TL40/T, available from Albright & Wilson) ; alkylether sulphates, such as sodium lauryl ether sulphate (eg. EMPICOL ESB70, available from Albright & Wilson) ; alkyl sulphonates, such as sodium alkane (C13-18) sulphonate (eg. HOSTAPUR SAS 30, available from Hoechst) ; alkylaryl sulphonates, such as sodium alkyl benzene sulphonate (eg. TEEPOL CM44, available from Shell) ; olefin sulphonates, such as sodium olefin (C5-18) sulphonate (eg. HOSTAPUR OS, available from Hoechst) ; acyl sarcosinates such as sodium lauryl sarcosinate (eg. HAMPOSYL L-95 available from Grace) ; - acyl taurides such as coconut methyl taurine (eg. FENOPEN TC 42, available from GAF); acyl isethionates such as sodium acyl isethionate [eg. JORDAPON Cl, available from Jordon] ; monoalkyi sulphosuccinates including sodium lauryl sulphosuccinate (eg. EMPICOL Si I, available from Albright & Wilson) , magnesium alkyl sulphosuccinate (eg. ELFANOL 616 Mg, available from AKZO) , sodium lauryl ethoxysulphosuccinate (eg. EMPICOL SDD, available from Albright & Wilson) , coconut monoethanolamide ethoxysulphosuccinate (eg. EMPICOL SGG) , disodium lauryl polyqlycolether sulphosuccinate (eg. SURTAGENE S30, available from CHEM-Y) , polyethyleneglycol sulphosuccinate (eg. REWOPOL SBFA 30, available from REWO) ; - dialkyl sulphosuccinates such as sodium dilauryl sulphosuccinate (eg. EMCOL 4500, available from Witco) ; acyl lactylates such as decanoyl lactylate (eg. PATIONIC 122a, available from Patterson, CJ) ; - acylated α-amino acids, such as sodium lauroyl glutamate (eg. ACYLGLUTAMATE LS-1, available from Ajinomoto Co. Inc) ; ethyl carboxylates , such as alkyl C12 -140 (EO) 4 OCH 2 C0 2 Na (eg. AKYPO RLM 38, available from Akzo) ; - monoalkyi phosphates and dialkyl phosphates, such as dioctyl phosphate Amphoteric surfactants, which are preferably chosen from: derivatives of aliphatic quaternary ammonium, phosphonium and sulphonium compounds, wherein the aliphatic radicals contain from 8 to 18 carbon atoms, and may be straight chain or branched, and further contain an anionic water-solubilising group, such as carboxyl, sulphonate, sulphate, phosphate or phosphonate. Preferred amphoteric surfactants include: Alkyl betaines such as lauryldimethyl betaine (eg. EMPIGEN BB, available from Albright & Wilson) ; Alkylamidopropyl betaines such as cocamidopropy betaine (eg. TEGOBETAIN L7, available from Goldschmidt) ; Alkylamphoqlycinates or Alkylamphopropionates such as cocoamphoglycinate (available from GAF) , and cocoamphopropionate; Sultaines such as cocamidopropylhydroxysultaine (eg. CYCLOTERIC BET-CS, available from Alcolac) . The most preferred amphoteric surfactants are lauryl dimethyl betaine and cocamidopropyl betaine. Such amphoteric surfactants can contribute to the foaming of the shampoo of the invention, while ameliorating the harshness of the anionic surfactant. Nonionic surfactants . e.g. comprising alkoxylated or glycosidic nonionic surfactants having an HLB of 8 or more. Above this value nonionics generally form clear isotropic solutions in combination with the other surfactants in the ranges defined above. Preferred nonionic surfactants are polyoxyethylene alkyl esters and polyoxyethylene alkyl ethers and alkyl polyglycosides. A suitable example of a polyoxyethylene alkyl ester is that having the CTFA designation Polysorbate 80, which is a mixture of oleate esters of sorbitol and sorbitol anhydrides, condensed with approximately 20 moles of ethylene oxide. Also suitable is Polysorbate 20 which is a mixture of laurate esters or sorbitol and sorbitol anhydrides condensed with approximately 20 moles of ethylene oxide. Polysorbate 80 and 20 are available commercially as TWEEN 80 and TWEEN 20 respectively from ICI Americas. Also suitable for use in the compositions of the invention is the polyethylene glycol ether of C9-11 alcohol with an average of 8 ethoxy units, which is available commerically as NONIDET LE-8T or as SYNPERONIC 91-8T, and the polyethylene glycol ether of C12-15 alcohol with an average of 9 ethoxy units which is available commerically as OOBANOL 25-9. Particularly useful alkyl polyglycosides include the glycosides of glucose or glucose oligomers where the alkyl chain can be C8-16 and the average number of glucose units is 1 to 2. A suitable example is ORAMIX NS10 which is the glucoside of C10-12 fatty alcohol with an average of about 1.5 glucose units. The hair growth promoters and other components of the composition, may be susceptible to microbial spoilage, particularly at the near neutral pH values. Therefore, the compositions according to the invention are preferably preserved in such a manner that they will enjoy a useful shelf life following manufacture and prior to sale and use. Ideally the compositions will have indefinite shelf life. In order to be preserved, the composition should preferably be made free, or substantially free, from viable microbial contaminants that are capable of resulting in microbial spoilage of the composition, and/or biodegradation of the hair growth promoter prior to topical application of the composition to mammalian skin or hair. Examples of methods that can be employed to achieve preservation of the composition, include the following: (i) Sterilisation The composition according to the invention can be preserved by sterilisation to remove or kill substantially all viable microbial contaminants. This can be achieved for example by irradiation using a lethal dose of gamma rays, by heat sterilisation or by ultrafiltration, using techniques that are well established in the art (ii) Chemical Preservation The composition according to the invention can also be preserved by including in it a chemical preservative which prevents the growth of, or kills, microorganisms. Examples of chemical preservatives include ethanol, benzoic acid, sodium benzoate, sorbic acid, potassium sorbate, sodium propionate and the methyl, ethyl, propyl and butyl esters of p-hydroxybenzoic acid. The amount of chemical preservative that can be incorporated in the composition according to the invention will generally be from 0.05 to 5%, preferably from 0.1 to 2% by weight, the amount chosen being sufficient to arrest microbial proliferation. (iii) Depressing water activity The composition according to the invention can also be preserved by the inclusion of a water activity depressant such as glycerol, propylene glycol, sorbitol, sugars and salts, for examples alkali metal halides, sulphates and carboxylates. When employing a water activity depressant, sufficient should be incorporated in the composition according to the invention to reduce the water activity (a") from 1 to < 0.9, preferably to < 0.85 and most preferably < 0.8, the lowest of these values being that at which yeasts, moulds and fungi will not proliferate. The hair growth promoter may be susceptable to hydrolysis, particularly when the pH value of the composition is highly alkaline. It is accordingly preferred that the composition, when aqueous, should have an acid to about neutral pH value. The preferred pH value of the composition, when aqueous, is from 2 to 9, preferably from 4 to <7, ideally f om 4 to 6.5. The invention also provides a process for the preparation of a composition suitable for topical application to mammalian skin or hair which comprises mixing a hair growth promoter as herein defined, with a suitable vehicle to provide a composition according to the invention, in which the hair growth promoter forms from 0.001 to 99% by weight of the composition, preferably from 0.01% to 20% by weight. The compositions of the invention can be formulated as liquids, for example as a lotion, shampoo, conditioner, milk or cream for use in conjunction with an applicator such as a roll-ball applicator, or a spray device such as an aerosol can containing propellant, or a container fitted with a pump to dispense the liquid product. Alternatively, the compositions of the invention can be solid or semi-solid, for example sticks, creams or gels, for use in conjunction with a suitable applicator or simply a tube, bottle or lidded jar, or as a liquid-impregnated fabric, such as a tissue wipe. The invention accordingly also provides a closed container containing a composition as herein defined. The invention also provides for a method of cosmetic treatment of the mammalian body and the use of hair growth promoter as herein defined for cosmetic purposes, i.e. for topical application to mammalian skin or hair for inducing, maintaining or increasing hair growth. The compositions according to the invention are primarily intended for topical application to the scalp of the human subject, particularly where the head is already bald or balding, in order to convert vellus hair to growth as terminal hair, or to increase the rate of growth of terminal hair. The compositions can also be applied profilactically to skalp and hair to reduce or prevent the onset of baldness. The amount of the composition and the frequency of application to the hair and/or scalp can vary widely, depending on personal needs, but it is suggested as an example that topical application of from 0.1 to 20g regularly, preferably from 0.1 to 5g, containing from 0.00001 to 5g (preferably at least O.Olg) of a hair growth promotor as herein defined, over the period of preferably six months or more will in most cases result in an improvement in hair growth. The application is preferably repeated weekly, more preferably twice weekly, particularly daily. Molecular weight determination Various different methods for determining the molecular weight distribution of protein hydrolysates and peptide mixtures are known in the art. The method preferably used for the purpose of this invention is Gel Permeation Chromatography, using as the column: Waters Protein-Pak 60, 7.8 mm * 300mm, with Protein-Pak 125 guard column, further using: 0.1 mol potassium phosphate buffer (pH 7), flow rate 1.0 ml/min, detection at 214 nm. The samples contained 0.2-0.3 mg/ml protein hydrolysate/peptide mixture in the K phosphate buffer. EVALUATION OF EFFICACY OF THE HAIR GROWTH PROMOTERS USING THE IN VITRO HAIR FOLLICLE GROWTH TEST The effect of compounds on hair growth may be assessed using the in vitro test described below which measures the elongation of isolated human hair follicles in a culture medium. Isolation of the hair follicle from skin This test includes the important step of isolating hair follicles having an undamaged hair bulb from human skin, for example, facelift skin, by microdissection. The critical step of separating the hair follicle with intact undamaged hair bulb from the subcutaneous fatty tissue in which it is situated accordingly involves severing the hair shaft of the follicle at a point below the epidermis of skin surface, so as to leave the hair bulb intact and undamaged while still bearing a portion of the hair shaft. Preferably, the hair shaft of the follicle is severed at the dermal-subcutaneous fat interface. Any suitable cutting instrument can be employed to sever the hair shaft in this manner, but a keratotome or a scalpel are preferred. The hair bulb with a hair shaft stump attached is then isolated from the skin by mechanically separating the hair from loosely adhering subcutaneous fat which normally surrounds the hair bulb. This is achieved after the dermis or upper layer of the skin has been separated and removed, to avoid damaging the hair bulb as it is pulled away. The hair bulb together with hair shaft stump attached, is then transferred in an otherwise undamaged and fully functioning, viable state to a nutrient medium. Culture of the isolated hair follicle The hair follicles isolated by the technique described herein are transferred to a suitable culture medium for subsequent testing of substances that can then influence their f ture development. The procedure now to be described represents a preferred method of culture and testing of hair growth. In accordance with the preferred method of culture, isolated hair follicles, are maintained in 1 ml of Williams E medium, either with or without a test hair growth substance, supplemented with antibodies (Penicillin and Streptomyein) , Insulin (lOng/ml) and Hydrocortisone (lOng/ml) . The medium was incubated at 37 C in an atmosphere of 5% CO + 95% air in individual wells of a 24 multiwell dish (Corning) , which permits detailed measurements to be made of the length of individual hair follicles. The medium was refreshed once during the experiment after 4 days. Williams E medium is available from FLOW Laboratory under Catalogue No. 12-502. The formula of Williams E medium is-described by Williams GM, et al., in Experimental Cell Research 69 (1971) on page 106. Daily growth rate and cumulative growth for each follicle were calculated by measuring the change in length of the follicles each day and from this the average of all the follicles was calculated. The response of an isolated hair follicle to a test substance, can accordingly be assessed by measuring the increase in length, if any, in the presence of a test substance against a control. EXAMPLES Example l This example illustrates a hair tonic which is suitable for topical application to hair or the scalp: % w/w HyPep 3401* 12 Ethanol 36 Water 52 Perfume q.s. * Enzymatic wheat gluten hydrolysate marketed by Quest International, Zwijndrecht, The Netherlands. Example 2 The following formulation represents a lotion which can be used topically in the treatment of balding heads: % w/w Ethanol 25 Butane-l,3-diol 34.4 Paramethyl benzoate 0.2 HyPep 4602* 8 N-acetyl-proline 0.6 Perfume 1 Water to 100 * Enzymatic wheat gluten hydrolysate marketed by Quest International, Zwijndrecht, The Netherlands. Example 3 The following example illustrates a lotiin according to the invention which can be applied topically to the scalp to prevent hair loss ans stimulate hair regrowth % w/w HyPep 4602 10 Glucaro-1,4-lactone 2 Ethanol 16 Citric acid 1.05 Water to 100 pH adjusted to 4.2 with sodium hydroxide. Example 4 The following example illustrates a shampoo for use in washing the hair and scalp and for promoting hair growth on the scalp: % w/w Sodium lauryl ether sulphate 41.4 (2 EO, 21% AD) Lauryl dimethylamino acetic acid betaine 4 (30% AD) Coconut fatty acid ethanol amine 1.5 Oleyl triethoxy phosphate (BRIPHOS 03D) 1 Polyglycol-polyamine condensation resin 1.5 (POLYQUART H, 50% active) Preservative, colouring matter, salt 0.58 HyPep 4602 20 Perf me q.s. Water to 100

Claims: CLAIMS 1. A composition, suitable for topical application to mammalian skin or hair, which comprises: i. glutamine containing peptides or peptide mixtures, in an amount effective to induce, increase and promote cell growth in mammalian skin, which have a molecular weight between 150 and 1000 Dalton, with the proviso that if the peptides are prepared synthetically glutamine is not at the acidic end of the peptide chain; ii. at least one cosmetically acceptable vehicle other than water. 2. A composition according to claim 1 wherein the total glutamine content is 5% by weight or more. 3. A composition according to claim 1 or 2 wherein the peptides are obtained by hydrolysis of protein. 4. A composition according to claim 3 wherein the protein is of vegetable origin. 5. A composition according to claim 4 wherein the protein is wheat gluten or soy protein 6. A composition according to claims 3-5 wherein the peptides are obtained by enzymatic hydrolysis. 7. A composition according to any one of claims 1-6 wherein the amount of peptide is between 0.001 and 99% by weight of the composition. 8. A composition according to claim 7 wherein the amount of peptide is between 0.01 and 20% by weight of the composition. 9. A composition according to any one of claims 1-8 wherein the cosmetically acceptable vehicle comprises one or more of emollients, propellants, solvents, humectants, powders. 10. A composition according to any one of claims 1-9, also comprising an activity enhancer. 11. A composition according to any one of claims 1-10, also comprising a surfactant. 12. Process for the preparation of a composition suitable for topical application to mammalian skin or hair which comprises mixing glutamine containing peptides or peptide mixtures which have a molecular weight between 150 and 1000 Dalton, with the proviso that if the peptides are prepared synthetically glutamine is not at the acidic end of the peptide chain, with at least one cosmetically acceptable vehicle other than water, to provide a composition in which the peptides form 0.001 - 99% by weight of the composition. 13. A process according to claim 12 in which the peptides form 0.01 - 20% by weight of the composition. 14. Method for cosmetic treatment of the mammalian body comprising applying to the skin or hair a composition according to any one of claims 1-11. 15. Method according to claim 14 wherein at least O.lg of the composition is regularly applied.

Inventors: DE LEVITA PAUL DAVID, VAN SCHIE BARTHOLOMEUS JOSEF, DE LEVITA, PAUL, DAVID, VAN SCHIE, BARTHOLOMEUS, JOSEF
Applicants: LEVITA PAUL DAVID DE, QUEST INT, SCHIE BARTHOLOMEUS JOSEF VAN, DE LEVITA, PAUL, DAVID, QUEST INTERNATIONAL B.V., VAN SCHIE, BARTHOLOMEUS, JOSEF
Application year: 1996
Main classification: A61K7
IPC codes: A61K8, A61K31, A61K38, A61P17, A61Q5, A61Q7