WO2022184763A1 - Use of galacto-oligosaccharides for providing skin care benefits - Google Patents

Use of galacto-oligosaccharides for providing skin care benefits Download PDF

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Publication number
WO2022184763A1
WO2022184763A1 PCT/EP2022/055253 EP2022055253W WO2022184763A1 WO 2022184763 A1 WO2022184763 A1 WO 2022184763A1 EP 2022055253 W EP2022055253 W EP 2022055253W WO 2022184763 A1 WO2022184763 A1 WO 2022184763A1
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human
acid
species
galacto
skin
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PCT/EP2022/055253
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French (fr)
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WO2022184763A8 (en
Inventor
Michael John Hoptroff
Joanne Elizabeth HUNT
Barry Murphy
Emily Grace Smith
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Unilever Ip Holdings B.V.
Unilever Global Ip Limited
Conopco, Inc., D/B/A Unilever
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Priority to EP22708949.7A priority Critical patent/EP4301329A1/en
Publication of WO2022184763A1 publication Critical patent/WO2022184763A1/en
Publication of WO2022184763A8 publication Critical patent/WO2022184763A8/en

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/005Preparations for sensitive skin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/60Sugars; Derivatives thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73Polysaccharides

Definitions

  • the present invention relates to the use of galacto-oligosaccharides for providing skin care benefits by supporting the growth of beneficial commensal microbial species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome.
  • An objective of the present invention is to provide skin care benefits by supporting the growth of beneficial commensal microbial species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome.
  • the invention provides the cosmetic use of galacto-oligosaccharides in a topical skin care composition as an active ingredient for providing skin care benefits by supporting the growth of beneficial commensal microbial species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome, characterised in that the beneficial commensal microbial species are Streptococcus species.
  • the invention also provides galacto-oligosaccharides, for use in supporting the growth of beneficial commensal Streptococcus species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome.
  • the invention also provides the use of galacto-oligosaccharides in, and for the manufacture of, topical compositions for supporting the growth of beneficial commensal Streptococcus species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome.
  • skin is understood as the layers which comprise it, from the uppermost layer or stratum corneum to the lowermost layer or hypodermis, both inclusive. These layers are composed of different types of cells such as keratinocytes, fibroblasts, melanocytes, mastocytes, neurons, and adipocytes. The term “skin” also comprises the scalp.
  • infant refers in general to a human baby or young child from birth up to about 5 years of age, such as from birth up to about 3 years of age and preferably from birth up to about 1 year of age.
  • Infant skin is structurally different to adult skin; the cells are smaller and the collagen fibres are thinner. According to clinical findings, infant stratum corneum is 30% thinner than in adults, and infant epidermis is 20% thinner than in adults.
  • the composition of infant skin is also different from that of adult skin, containing less natural moisturising factors (NMFs), less lipids and less melanin. Infant skin also has a higher pH than adult skin.
  • NMFs moisturising factors
  • Infant skin also has a higher pH than adult skin.
  • skin care means regulating and/or improving cosmetic qualities of the skin, (as opposed to curing, treating, or preventing a disease or disorder). Accordingly, such cosmetic qualities are subject to regulation and/or improvement both in healthy subjects as well as those which present diseases or disorders of the skin (such as psoriasis, lichen planus, folliculitis or atopic dermatitis).
  • Examples of skin care benefits in the context of this invention include providing a smoother, more even texture; improving the elasticity or resiliency of the skin; improving the firmness of the skin; improving the hydration status or moisturization of the skin; improving skin barrier properties; and reducing the appearance of redness or skin blotches.
  • galacto-oligosaccharides are used for supporting the growth of beneficial commensal Streptococcus species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome.
  • VGS beneficial commensal Streptococcus species in the context of this invention are bacteria of the viridans group of streptococci (VGS).
  • VGS are a group of catalase negative, Gram-positive cocci with a chaining morphology on microscopic examination. They are leucine aminopeptidase positive, pyrrolidonylarylamidase negative, and do not grow in 6.5% NaCI, and almost all species are negative for growth on bile esculin agar.
  • VGS can be differentiated from Streptococcus pneumoniae in that they are optochin-resistant and are not bile soluble; they also lack either the polysaccharide-based capsule typical of S.
  • VGS ulcerative colitis
  • Streptococcus species in the context of this invention are the salivarius group of VGS.
  • the salivarius group of VGS consists of three genetically similar species: Streptococcus salivarius, Streptococcus vestibularis and Streptococcus thermophilus (also known as Streptococcus salivarius subsp. thermophilus).
  • S.thermophilus is classified as a lactic acid bacterium, and is believed to have developed separately from pathogenic Streptococcus species for at least 3000 years. Its genome sequence does not contain any of the virulence regions common to pathogenic species of Streptococcus.
  • Streptococcus thermophilus has been observed to increase the level of ceramides in stratum corneum in vitro and in vivo. Ceramides are the major lipid constituent of lamellar sheets present in the intercellular spaces of the stratum corneum, which are thought to provide the barrier property of the epidermis. Studies have shown that topical administration of Streptococcus thermophilus cream in patients with atopic dermatitis led to a significant improvement in erythema, scaling and pruritus.
  • Streptococcus thermophilus can inhibit Staphylococcus aureus growth by producing hydrogen peroxide; and certain strains of Streptococcus thermophilus have been shown to produce exosaccharide polymers including hyaluronic acid.
  • Galacto-oligosaccharides are composed of oligosaccharide chains having from 2 to 10 saccharide units which contain at least two adjacent galactose units linked by galactosyl- galactose glycosidic bonds. They can be classified into distinct groups depending on the type of glycosidic bonds present. Plant based GOS are a-GOS whereas GOS prepared from lactose are b-GOS. In either instance, the products obtained usually consist of a mixture of GOS molecules of varying DP, with chain size and size distribution depending on the source, the harvesting conditions (where applicable) and the processing conditions.
  • High performance anion exchange chromatography with pulsed amperometric detection can be used to determine the number average degree of polymerization (DP n ) .
  • a-GOS also known as the raffinose family of oligosaccharides
  • a-GOS can be obtained by extraction from the seeds of legumes such as soybean (Glycine max), lupin (Lupinus albus), lentil ( Lens culinaris), chickpea (Cicer arietinum) and pea ( Pisum sativum).
  • Examples of such materials include a(1 6) galactosides with a DP ranging from 3 to 7 saccharide units, such as stachyose (a-D-galactopyranosyl-(1 6)-a-D-galactopyranosyl-(1 6)-a-D- glucopyranosyl-(1 2) ⁇ -D-fructofuranoside); verbascose (a-D-galactopyranosyl-(1 6)-[a- D-galactopyranosyl-(1 6)] 2 -a-D-glucopyranosyl-(1 2) ⁇ -D-fructofuranoside); and ajugose (a-D-galactopyranosyl-(1 6)-[a-D-galactopyranosyl-(1 6)] 3 -a-D-glucopyranosyl-(1 2)-b- D-f ru ctof
  • b-GOS also known as oligogalactosyllactose, oligogalactose, oligolactose, transgalactosylated oligosaccharide, and fransgalacto-oligosaccharide
  • b-GOS also known as oligogalactosyllactose, oligogalactose, oligolactose, transgalactosylated oligosaccharide, and fransgalacto-oligosaccharide
  • K the b-galactosidase from K.
  • lactis produces predominantly b-(1 6)-linked GOS; the b-galactosidase from Aspergillus oryzae produces mainly b-(1 3) and b-(1 6) linkages; Bacillus circulans b-galactosidase forms mainly b- (1 4)-linked GOS; whereas b-galactosidases from Lactobacillus spp. preferably form b- (1 6) and b-(1 3) linkages.
  • the resulting b-GOS are typically composed of a chain of b- D-galactose residues terminating at the reducing end with a D-glucose residue; and generally containing between 2 and 8 monosaccharide residues.
  • the chains are predominantly linear, but a few branched structures have been reported.
  • the reducing end is galactose or fructose.
  • compositions for use in the invention can be formulated in a variety of forms for topical application, and will generally contain from about 0.01 to about 10%, preferably from about 0.1 to about 5% of galacto-oligosaccharide (by weight based on the total weight of the composition).
  • Topical compositions for use in the invention will generally include a cosmetically acceptable vehicle.
  • Cosmetically acceptable means that the vehicle is suitable for topical application to the skin, has good aesthetic properties, is compatible with the galacto- oligosaccharide and any other ingredients, and will not cause any safety or toxicity concerns.
  • the vehicle may comprise an aqueous phase, an oil phase, an alcohol, a silicone phase, or a mixture thereof, and may be in the form of an emulsion.
  • Emulsions can have a range of consistencies including thin lotions (which may also be suitable for spray or aerosol delivery), creamy lotions, light creams, and heavy creams.
  • Exemplary emulsions include water-in-oil emulsions, oil-in-water emulsions, silicone-in-water emulsions, water-in-silicone emulsions, polyol-in-silicone emulsions, silicone-in-polyol emulsions, polyol-in-oil emulsions, oil-in-polyol emulsions, wax-in-water emulsions and water-oil-water triple emulsions.
  • Preferred emulsions include oil-in-water emulsions and water-in-oil emulsions.
  • Topical cosmetic compositions in the form of an emulsion, and suitable for use in the invention typically have an oil phase containing at one or more cosmetically acceptable fatty materials which may be liquid or solid at room temperature (25°C).
  • Suitable cosmetically acceptable fatty materials include naturally derived oils (such as sunflower oil, borage oil, soybean oil, castor oil, olive oil and almond oil); esters of monoalcohols or of glycols with monocarboxylic or polycarboxylic acids, at least one of the alcohols and/or acids comprising at least one hydrocarbon-based chain containing at least 6 carbon atoms (such as octyl palmitate, isopropyl myristate, isopropyl palmitate, isopropyl isostearate, hexyl laurate, isohexyl laurate, isohexyl palmitate, decyl oleate, isodecyl oleate, hexadecyl stearate, decyl stearate, dihexyldecyl adipate, lauryl lactate, myristyl lactate, cetyl lactate, oleyl stearate, oleyl oleate
  • Topical cosmetic compositions in the form of an emulsion, and suitable for use in the invention typically have an aqueous phase, with the amount of water in such an emulsion suitably ranging from about 5 to about 95%, preferably from about 35 to about 80% (by weight based on the total weight of the composition).
  • the aqueous phase may also include one or more organic liquids that are miscible with water at room temperature (25°C).
  • Exemplary water-miscible organic liquids include monohydric and polyhydric alcohols and derivatives thereof such as C2-C6 alkanols (such as ethanol and isopropanol); C2-C10 glycols and polyols (such as glycerol, propylene glycol, butylene glycol, pentylene glycol, hexylene glycol, caprylyl glycol, dipropylene glycol, and diethylene glycol); C3-C16 glycol ethers (such as mono-, di-, or tripropylene glycol (C1-C4) alkyl ethers and mono-, di-, or triethylene glycol (C1-C4) alkyl ethers) and polyethylene glycol having 2 to 12 oxyethylene units.
  • C2-C6 alkanols such as ethanol and isopropanol
  • C2-C10 glycols and polyols such as glycerol, propylene glycol, butylene glycol, pentylene
  • Topical cosmetic compositions in the form of an emulsion, and suitable for use in the invention generally include emulsifiers and solubilizers, to enable two or more immiscible components to be combined homogeneously and to help stabilize the composition.
  • the amount of emulsifier and solubilizer in such an emulsion suitably ranges from about 0.1 to about 30%, preferably from about 1 to about 8% (by weight based on the total weight of the composition).
  • Emulsifiers that may be used to form O/W or W/O emulsions include sorbitan oleate, sorbitan sesquioleate, sorbitan isostearate, sorbitan trioleate, PEG-20 sorbitan isostearate, polyglyceryl-3-diisostearate, polyglycerol esters of oleic/isostearic acid, polyglyceryl-6 hexaricinolate, polyglyceryl-4-oleate, polyglyceryl-4 oleate/PEG-8 propylene glycol cocoate, polyglyceryl-2 dipolyhydroxystearate, PEG-30 dipolyhydroxystearate, oleamide DEA, TEA myristate, TEA stearate, magnesium stearate, sodium stearate, potassium laurate, potassium ricinoleate, sodium cocoate, sodium tallowate, potassium castorate, sodium oleate, cetyl phosphate, di
  • Topical cosmetic compositions suitable for use in the invention may also take the form of a skin cleanser incorporating one or more cleansing surfactants which, when combined with water and mechanically agitated, generate a foam or lather.
  • the amount of cleansing surfactant suitably ranges from about 5 to about 40%, preferably from about 10 to about 35% (by weight based on the total weight of the composition).
  • Exemplary cleansing surfactants include anionic surfactants such as ammonium lauroyl sarcosinate, sodium trideceth sulfate, sodium lauroyl sarcosinate, sodium myristoyl sarcosinate, ammonium laureth sulfate, sodium laureth sulfate, ammonium lauryl sulfate, sodium lauryl sulfate, ammonium cocoyl isethionate, sodium cocoyl isethionate, sodium lauroyl isethionate, sodium cocoyl glycinate, sodium lauroyl glycinate, sodium lauroyl taurate, sodium methyl lauroyl taurate, sodium methyl oleoyl taurate, sodium cetyl sulfate, sodium lauroyl lactylate and triethanolamine lauroyl lactylate and mixtures thereof; nonionic surfactants such as lauramine oxide, cocoamine oxide, decyl polyglucose,
  • Topical cosmetic compositions for use in the invention may also be formulated in a single phase carrier such as water and/or one or more water miscible organic liquids (such as the monohydric and polyhydric alcohols and derivatives thereof described above).
  • a single phase carrier such as water and/or one or more water miscible organic liquids (such as the monohydric and polyhydric alcohols and derivatives thereof described above).
  • Topical cosmetic compositions for use in the invention may also be formulated in solid forms such as gels or sticks.
  • a topical cosmetic composition for use in the invention may include additional skin care actives (for improving the physical and/or aesthetic characteristics of the skin).
  • additional skin care actives which are suitable for use in the invention include vitamins, minerals and/or antioxidants, emollients, humectants, skin anti-hyperpigmentation agents, sunscreens, anti-irritants, exfoliating agents and mixtures thereof.
  • Vitamins, minerals and/or antioxidants suitable for use in the invention include natural botanical antioxidants derived from plant materials such as fruits, vegetables, herbs and spices (such as goji berry, white tea, rosemary, green tea, grape seed and lemongrass extracts); vitamin A and its precursors or derivatives (such as beta-carotene, retinyl palmitate); vitamin B3 and its precursors or derivatives (such as niacinamide); vitamin B5 and its precursors or derivatives (such as panthenol and its precursors or derivatives); vitamin C and its precursors or derivatives (such as tetrahexyldecyl ascorbate, ascorbyl palmitate); vitamin E and its precursors or derivatives (such as d-alpha-tocopherol, tocopheryl acetate); vitamin K and its precursors or derivatives; selenium and its derivatives (such as L-selenomethionine); and alpha lipoic acid.
  • natural botanical antioxidants derived from plant materials such as fruits, vegetables,
  • Emollients suitable for use in the invention act to increase and maintain moisture in the skin by providing a protective coating to impede epidermal water loss.
  • emollients include C10-20 fatty alcohols and acids (such as cetyl, myristyl, palmitic and stearyl alcohols and acids); and C10-40 hydrocarbons (such as mineral oil, petroleum jelly, squalene and isoparaffins).
  • Humectants suitable for use in the invention act to increase and maintain moisture in the skin by attracting water to the stratum corneum of the epidermis.
  • humectants include amino acids, chondroitin sulfate, glycerin, diglycerin, triglycerin, polyglycerin, polypropylene glycol, polyethylene glycol, ethylene glycol, diethylene glycol, triethylene glycol, propylene glycol, dipropylene glycol, hexylene glycol, 1,3-butylene glycol, 1,4- butylene glycol, ethylene glycol monoalkyl ether, diethylene glycol monoalkyl ether, erythritol, fructose, glucose, maltose, sucrose, lactose, xylose, inositol, lactitol, xylitol, sorbitol, mannitol, maltitol, panthenol, pentaeryth
  • Skin anti-hyperpigmentation agents suitable for use in the invention include natural botanical agents derived from plant materials (such as Arctostaphylos patula and Arctostaphylos viscida extracts, Emblica officinalis extract, Mitracarpus scaber extract, Uva ursi (bearberry) extract, Morns bombycis (mulberry) extract, Morns alba (white mulberry) extract, Broussonetia papyrifera (paper mulberry) extract, licorice extract, acerola cherry extract, Chlorella vulgaris extract, Aloe ferrox extract and Rumex occidentalis extract); synthetic or natural sugar amines (such as glucosamine, N-acetyl glucosamine, glucosamine sulfate, mannosamine, N-acetyl mannosamine, galactosamine, N-acetyl galactosamine and their hydrochloride salts); retinoids (such as retinol, retinal
  • Sunscreens suitable for use in the invention protect the skin from ultraviolet (UV) solar radiation falling within both the UVB region (between 290nm to 320 nm wavelengths) and the UVA region (between 320nm and 400nm wavelengths).
  • sunscreens include methoxycinnamate derivatives (such as octyl methoxycinnamate and isoamyl methoxycinnamate); camphor derivatives (such as 4-methyl benzylidene camphor, camphor benzalkonium methosulfate, and terephthalylidene dicamphor sulfonic acid); salicylate derivatives (such as octyl salicylate and homosalate); sulfonic acid derivatives (such as phenylbenzimidazole sulfonic acid); benzone derivatives (such as dioxybenzone, sulisobenzone, and oxybenzone); benzoic acid derivatives (such as aminobenzoic acid and octyld
  • Anti-irritants suitable for use in the invention include allantoin, aloe vera, a-bisabolol, caffeine, chamomile extract, Cola nitada extract, cucumber extract, dipotassium glycyrrhizinate, glycyrrhizic acid, green tea extract, lecithin or hydrogenated lecithin, licorice extract, Avena sativa (oat) meal extract, tea tree oil, salicylic acid, acetylsalicylic acid, strontium acetate, strontium chloride, strontium nitrate, fatty acids with anti-irritant properties (such as linoleic acid and linolenic acid) and aromatic aldehydes with anti-irritant properties (such as 4-methoxy benzaldehyde, 4-ethoxy benzaldehyde, 4-butoxy benzaldehyde and 4- pentoxy benzaldehyde).
  • Exfoliating agents suitable for use in the invention include benzoyl peroxide, benzoic acid, 3- hydroxy benzoic acid, salicylic acid, acetic acid, trichloroacetic acid, 1-pyrrolidone-5- carboxylic acid, a-hydroxy acids (such as glycolic acid, lactic acid, malic acid, tartaric acid, and citric acid); b-hydroxy acids (such as b-hydroxybutyric acid); a-keto acids (such as pyruvic acid, 2-oxopropanoic acid, 2-oxobutanoic acid and 2-oxopentanoic acid); and oxa acids (such as 3,6,9-trioxaundecanedioic acid).
  • a-hydroxy acids such as glycolic acid, lactic acid, malic acid, tartaric acid, and citric acid
  • b-hydroxy acids such as b-hydroxybutyric acid
  • a-keto acids such as pyruvic acid, 2-oxopropanoic
  • a topical cosmetic composition for use in the invention may include additional functional ingredients (for improving the physical and/or aesthetic characteristics of the composition).
  • water soluble or colloidally water soluble polymeric thickening agents such as hydroxyethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, polyquaternium-10, carrageenan, guar gum, hydroxypropyl guar gum, xanthan gum, polyvinylalcohol, acrylic acid/ethyl acrylate copolymers, carboxyvinyl polymers, cross-linked polyacrylate polymers and polyacrylamide polymers
  • structurant clays such as magnesium aluminum silicate, attapulgite, bentonite, montmorillonite and hectorite
  • inorganic pigments such as titanium oxide, zirconium oxide, cerium oxide zinc oxide, iron oxide, chromium oxide and ferric blue
  • organic pigments such as carbon black and barium, strontium, calcium, and aluminium lakes
  • pearlescent agents such as mica coated with titanium oxide and/or iron oxide
  • dyes, preservatives such as hydroxyethyl cellulose, methyl
  • a topical cosmetic composition for use in the invention may be packaged in a suitable container to suit its viscosity and intended use by the consumer.
  • a lotion or a cream can be packaged in a bottle or a roll-ball applicator, or a propellant-driven aerosol device or a container fitted with a pump suitable for finger operation.
  • the composition When the composition is a cream, it can simply be stored in a non-deformable bottle or squeeze container, such as a tube or a lidded jar.
  • the composition is suitably applied to the skin, preferably infant skin, at the rate of one or two applications per day.
  • an amount corresponding to about 1 to 2 ml of the composition per application is applied uniformly over the area of treatment twice daily for a period of at least 7 (seven) days, more preferably at least 30 (thirty) days.
  • the invention will be further illustrated by the following, non-limiting Examples.
  • GOS Galacto-oligosaccharide
  • Test bacterial inoculums were prepared in culture media for 18 hours at 37 °C under C0 2 conditions. Inoculums were centrifuged and resuspended in liquid media diluted to 50% in sterile distilled water. The inoculums were adjusted to a cell density of 1.5-5 x 10 s CFU/ml.
  • microtiter plate was incubated without shaking at 37 °C for 24 hours and an endpoint optical density read taken at 620 nm.
  • the HydroPlate® was read kinetically for fluorescence for 24 hours at 37°C.
  • PreSens calibration software pHSolver_v10 was used as part of the analysis.
  • Table 1 shows the endpoint optical density of the tested bacterial species after incubation with GOS, compared to the growth control.
  • Table 2 The following Table 3 shows the endpoint optical density and calculated acid produced when S. thermophilus (ATCC® 19258TM) is incubated with different concentrations of GOS, compared to the growth control.

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Abstract

The invention provides the cosmetic use of galacto-oligosaccharides in a topical skin care composition as an active ingredient for providing skin care benefits by supporting the growth of beneficial commensal microbial species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome, characterised in that the beneficial commensal microbial species are Streptococcus species.

Description

USE OF GALACTO-OLIGOSACCHARIDES FOR PROVIDING SKIN CARE BENEFITS
Field of the Invention
The present invention relates to the use of galacto-oligosaccharides for providing skin care benefits by supporting the growth of beneficial commensal microbial species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome.
Background of the Invention
Birth marks a time of drastic change for the skin of the newborn as it undergoes a sudden transition from an almost bacteria-free aqueous environment to an atmospheric one with constant exposure to microbes. Immediately after birth, infant skin is colonized by surrounding microorganisms. By 4 to 6 weeks after birth, the infant cutaneous microbiome has already significantly expanded and diversified. The microbiome stabilizes over time and becomes progressively site specific. This process begins within the first 3 months of life and evolves over the first year, reflecting a balance between the influx of microbial species and niche selection.
Evidence suggests that the timely and proper establishment of a healthy cutaneous microbiome in infancy is crucial in promoting normal epithelial development and integrity, as well as shaping immune responses and protecting against potential pathogens. This is especially important since infant skin is more permeable, more prone to dryness and more vulnerable to the environment than adult skin. If not properly cared for, it may also become prone to inflammatory conditions such as atopic dermatitis and nappy dermatitis, or even infections such as candidiasis.
Studies show that the healthy cutaneous microbiome is significantly more diverse in young children than in teenagers or adults; and that skin microorganisms may be clustered into adult-associated and childhood-associated groups. Streptococcus species have a significantly greater relative abundance in young children, whereas lipophilic Cutibacterium and Corynebacterium replace Streptococcus after puberty and become dominant in adulthood. An objective of the present invention is to provide skin care benefits by supporting the growth of beneficial commensal microbial species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome.
Summary of the Invention
The invention provides the cosmetic use of galacto-oligosaccharides in a topical skin care composition as an active ingredient for providing skin care benefits by supporting the growth of beneficial commensal microbial species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome, characterised in that the beneficial commensal microbial species are Streptococcus species.
The invention also provides galacto-oligosaccharides, for use in supporting the growth of beneficial commensal Streptococcus species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome.
The invention also provides the use of galacto-oligosaccharides in, and for the manufacture of, topical compositions for supporting the growth of beneficial commensal Streptococcus species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome.
Detailed Description and Preferred Embodiments
In the context of this invention, “skin” is understood as the layers which comprise it, from the uppermost layer or stratum corneum to the lowermost layer or hypodermis, both inclusive. These layers are composed of different types of cells such as keratinocytes, fibroblasts, melanocytes, mastocytes, neurons, and adipocytes. The term “skin” also comprises the scalp.
As used herein, the term “infant” refers in general to a human baby or young child from birth up to about 5 years of age, such as from birth up to about 3 years of age and preferably from birth up to about 1 year of age. Infant skin is structurally different to adult skin; the cells are smaller and the collagen fibres are thinner. According to clinical findings, infant stratum corneum is 30% thinner than in adults, and infant epidermis is 20% thinner than in adults. The composition of infant skin is also different from that of adult skin, containing less natural moisturising factors (NMFs), less lipids and less melanin. Infant skin also has a higher pH than adult skin. These differences in structure and composition lead to differences in the function of the skin as a barrier. One of the main measurements of skin barrier is the rate of transepidermal water loss (TEWL).
Infant skin has a higher rate of TEWL than adult skin. Studies show that TEWL rates are highest in younger infant age groups (3 to 6 months old) and continue to show higher levels than adult skin through the first year of life.
As used herein, “skin care" means regulating and/or improving cosmetic qualities of the skin, (as opposed to curing, treating, or preventing a disease or disorder). Accordingly, such cosmetic qualities are subject to regulation and/or improvement both in healthy subjects as well as those which present diseases or disorders of the skin (such as psoriasis, lichen planus, folliculitis or atopic dermatitis).
Examples of skin care benefits in the context of this invention include providing a smoother, more even texture; improving the elasticity or resiliency of the skin; improving the firmness of the skin; improving the hydration status or moisturization of the skin; improving skin barrier properties; and reducing the appearance of redness or skin blotches.
According to this invention, galacto-oligosaccharides are used for supporting the growth of beneficial commensal Streptococcus species which are resident in the human cutaneous microbiome, in particular the human infant cutaneous microbiome.
Examples of beneficial commensal Streptococcus species in the context of this invention are bacteria of the viridans group of streptococci (VGS). The VGS are a group of catalase negative, Gram-positive cocci with a chaining morphology on microscopic examination. They are leucine aminopeptidase positive, pyrrolidonylarylamidase negative, and do not grow in 6.5% NaCI, and almost all species are negative for growth on bile esculin agar. VGS can be differentiated from Streptococcus pneumoniae in that they are optochin-resistant and are not bile soluble; they also lack either the polysaccharide-based capsule typical of S. pneumoniae or the Lancefield antigens of the pyogenic members of the genus. The existence of VGS is considered commensal in nature, where their presence and physiology leads to acidification of their immediate environment, thus making colonization and subsequent infection of such sites difficult by other pathogens.
Particularly beneficial Streptococcus species in the context of this invention are the salivarius group of VGS. The salivarius group of VGS consists of three genetically similar species: Streptococcus salivarius, Streptococcus vestibularis and Streptococcus thermophilus (also known as Streptococcus salivarius subsp. thermophilus). S.thermophilus is classified as a lactic acid bacterium, and is believed to have developed separately from pathogenic Streptococcus species for at least 3000 years. Its genome sequence does not contain any of the virulence regions common to pathogenic species of Streptococcus. Streptococcus thermophilus has been observed to increase the level of ceramides in stratum corneum in vitro and in vivo. Ceramides are the major lipid constituent of lamellar sheets present in the intercellular spaces of the stratum corneum, which are thought to provide the barrier property of the epidermis. Studies have shown that topical administration of Streptococcus thermophilus cream in patients with atopic dermatitis led to a significant improvement in erythema, scaling and pruritus. It has also been suggested that Streptococcus thermophilus can inhibit Staphylococcus aureus growth by producing hydrogen peroxide; and certain strains of Streptococcus thermophilus have been shown to produce exosaccharide polymers including hyaluronic acid.
Galacto-oligosaccharides
Galacto-oligosaccharides (GOS) are composed of oligosaccharide chains having from 2 to 10 saccharide units which contain at least two adjacent galactose units linked by galactosyl- galactose glycosidic bonds. They can be classified into distinct groups depending on the type of glycosidic bonds present. Plant based GOS are a-GOS whereas GOS prepared from lactose are b-GOS. In either instance, the products obtained usually consist of a mixture of GOS molecules of varying DP, with chain size and size distribution depending on the source, the harvesting conditions (where applicable) and the processing conditions. High performance anion exchange chromatography (HPAEC) with pulsed amperometric detection can be used to determine the number average degree of polymerization (DPn) . a-GOS (also known as the raffinose family of oligosaccharides) can be obtained by extraction from the seeds of legumes such as soybean (Glycine max), lupin (Lupinus albus), lentil ( Lens culinaris), chickpea (Cicer arietinum) and pea ( Pisum sativum). Examples of such materials include a(1 6) galactosides with a DP ranging from 3 to 7 saccharide units, such as stachyose (a-D-galactopyranosyl-(1 6)-a-D-galactopyranosyl-(1 6)-a-D- glucopyranosyl-(1 2)^-D-fructofuranoside); verbascose (a-D-galactopyranosyl-(1 6)-[a- D-galactopyranosyl-(1 6)]2-a-D-glucopyranosyl-(1 2)^-D-fructofuranoside); and ajugose (a-D-galactopyranosyl-(1 6)-[a-D-galactopyranosyl-(1 6)]3-a-D-glucopyranosyl-(1 2)-b- D-f ru ctof u ran os i d e) . b-GOS (also known as oligogalactosyllactose, oligogalactose, oligolactose, transgalactosylated oligosaccharide, and fransgalacto-oligosaccharide) occur naturally in the milk of many animals, including humans, cows and wallabies. However, most commercial b- GOS are synthesized from lactose through transgalactosylation reactions, using b- galactosidases as biocatalysts under conditions which favour transgalactosylation. Different species possess different specificities for building glycosidic linkages and therefore produce different GOS mixtures. For example, the b-galactosidase from K. lactis produces predominantly b-(1 6)-linked GOS; the b-galactosidase from Aspergillus oryzae produces mainly b-(1 3) and b-(1 6) linkages; Bacillus circulans b-galactosidase forms mainly b- (1 4)-linked GOS; whereas b-galactosidases from Lactobacillus spp. preferably form b- (1 6) and b-(1 3) linkages. The resulting b-GOS are typically composed of a chain of b- D-galactose residues terminating at the reducing end with a D-glucose residue; and generally containing between 2 and 8 monosaccharide residues. The chains are predominantly linear, but a few branched structures have been reported. In some instances, the reducing end is galactose or fructose.
Mixtures of any of the above described materials may also be used.
Product Forms
Compositions for use in the invention can be formulated in a variety of forms for topical application, and will generally contain from about 0.01 to about 10%, preferably from about 0.1 to about 5% of galacto-oligosaccharide (by weight based on the total weight of the composition).
Topical compositions for use in the invention will generally include a cosmetically acceptable vehicle. The term “cosmetically acceptable” means that the vehicle is suitable for topical application to the skin, has good aesthetic properties, is compatible with the galacto- oligosaccharide and any other ingredients, and will not cause any safety or toxicity concerns.
The vehicle may comprise an aqueous phase, an oil phase, an alcohol, a silicone phase, or a mixture thereof, and may be in the form of an emulsion. Emulsions can have a range of consistencies including thin lotions (which may also be suitable for spray or aerosol delivery), creamy lotions, light creams, and heavy creams.
Exemplary emulsions include water-in-oil emulsions, oil-in-water emulsions, silicone-in-water emulsions, water-in-silicone emulsions, polyol-in-silicone emulsions, silicone-in-polyol emulsions, polyol-in-oil emulsions, oil-in-polyol emulsions, wax-in-water emulsions and water-oil-water triple emulsions. Preferred emulsions include oil-in-water emulsions and water-in-oil emulsions.
Topical cosmetic compositions in the form of an emulsion, and suitable for use in the invention, typically have an oil phase containing at one or more cosmetically acceptable fatty materials which may be liquid or solid at room temperature (25°C).
Suitable cosmetically acceptable fatty materials include naturally derived oils (such as sunflower oil, borage oil, soybean oil, castor oil, olive oil and almond oil); esters of monoalcohols or of glycols with monocarboxylic or polycarboxylic acids, at least one of the alcohols and/or acids comprising at least one hydrocarbon-based chain containing at least 6 carbon atoms (such as octyl palmitate, isopropyl myristate, isopropyl palmitate, isopropyl isostearate, hexyl laurate, isohexyl laurate, isohexyl palmitate, decyl oleate, isodecyl oleate, hexadecyl stearate, decyl stearate, dihexyldecyl adipate, lauryl lactate, myristyl lactate, cetyl lactate, oleyl stearate, oleyl oleate, oleyl myristate, lauryl acetate, cetyl propionate, isononyl isononanoate, propylene glycol dicaprate, diisopropyl adipate, dibutyl adipate, and oleyl adipate); ethers (such as dicapryl ether); fatty alcohols (such as cetyl alcohol, stearyl alcohol and behenyl alcohol); isoparaffins (such as isooctane, isododecane and isohexadecane); silicone oils (such as dimethicones, cyclic silicones, and polysiloxanes); and hydrocarbon oils (such as mineral oil, petrolatum and polyisobutene); fatty acids containing from 8 to 30 carbon atoms, (such as stearic acid, lauric acid, palmitic acid and oleic acid); vegetable fats (such as cocoa butter, coconut oil, palm oil and shea butter) ; natural or synthetic waxes (such as lanolin wax, beeswax, carnauba wax, candelilla wax, paraffin wax, lignite wax, microcrystalline waxes, ceresin, ozokerite, and polyethylene waxes), hydrogenated oils which are solid at 25° C (such as hydrogenated castor oil, hydrogenated jojoba oil, hydrogenated palm oil, hydrogenated tallow and hydrogenated coconut oil) and fatty esters that are solid at 25°C (such as C20-40 alkyl stearate).
Mixtures of any of the above described materials may also be used.
Topical cosmetic compositions in the form of an emulsion, and suitable for use in the invention, typically have an aqueous phase, with the amount of water in such an emulsion suitably ranging from about 5 to about 95%, preferably from about 35 to about 80% (by weight based on the total weight of the composition). The aqueous phase may also include one or more organic liquids that are miscible with water at room temperature (25°C). Exemplary water-miscible organic liquids include monohydric and polyhydric alcohols and derivatives thereof such as C2-C6 alkanols (such as ethanol and isopropanol); C2-C10 glycols and polyols (such as glycerol, propylene glycol, butylene glycol, pentylene glycol, hexylene glycol, caprylyl glycol, dipropylene glycol, and diethylene glycol); C3-C16 glycol ethers (such as mono-, di-, or tripropylene glycol (C1-C4) alkyl ethers and mono-, di-, or triethylene glycol (C1-C4) alkyl ethers) and polyethylene glycol having 2 to 12 oxyethylene units.
Topical cosmetic compositions in the form of an emulsion, and suitable for use in the invention, generally include emulsifiers and solubilizers, to enable two or more immiscible components to be combined homogeneously and to help stabilize the composition. The amount of emulsifier and solubilizer in such an emulsion suitably ranges from about 0.1 to about 30%, preferably from about 1 to about 8% (by weight based on the total weight of the composition). Emulsifiers that may be used to form O/W or W/O emulsions include sorbitan oleate, sorbitan sesquioleate, sorbitan isostearate, sorbitan trioleate, PEG-20 sorbitan isostearate, polyglyceryl-3-diisostearate, polyglycerol esters of oleic/isostearic acid, polyglyceryl-6 hexaricinolate, polyglyceryl-4-oleate, polyglyceryl-4 oleate/PEG-8 propylene glycol cocoate, polyglyceryl-2 dipolyhydroxystearate, PEG-30 dipolyhydroxystearate, oleamide DEA, TEA myristate, TEA stearate, magnesium stearate, sodium stearate, potassium laurate, potassium ricinoleate, sodium cocoate, sodium tallowate, potassium castorate, sodium oleate, cetyl phosphate, diethanolamine cetyl phosphate, potassium cetyl phosphate, sodium glyceryl oleate phosphate, dimethicone copolyol, cetyl dimethicone copolyol, octyldimethicone ethoxyglucoside copolyol, dimethicone copolyol crosspolymer and laurylmethicone copolyol. Mixtures of any of the above described materials may also be used.
Topical cosmetic compositions suitable for use in the invention may also take the form of a skin cleanser incorporating one or more cleansing surfactants which, when combined with water and mechanically agitated, generate a foam or lather. The amount of cleansing surfactant suitably ranges from about 5 to about 40%, preferably from about 10 to about 35% (by weight based on the total weight of the composition). Exemplary cleansing surfactants include anionic surfactants such as ammonium lauroyl sarcosinate, sodium trideceth sulfate, sodium lauroyl sarcosinate, sodium myristoyl sarcosinate, ammonium laureth sulfate, sodium laureth sulfate, ammonium lauryl sulfate, sodium lauryl sulfate, ammonium cocoyl isethionate, sodium cocoyl isethionate, sodium lauroyl isethionate, sodium cocoyl glycinate, sodium lauroyl glycinate, sodium methyl cocoyl taurate, sodium methyl lauroyl taurate, sodium methyl oleoyl taurate, sodium cetyl sulfate, sodium lauroyl lactylate and triethanolamine lauroyl lactylate and mixtures thereof; nonionic surfactants such as lauramine oxide, cocoamine oxide, decyl polyglucose, lauryl polyglucose, sucrose cocoate and sucrose laurate and mixtures thereof; and amphoteric surfactants such as disodium lauroamphodiacetate, sodium lauroamphoacetate, cetyl dimethyl betaine, cocoamidopropyl betaine and cocoamidopropyl hydroxy sultaine and mixtures thereof.
Mixtures of any of the above described materials may also be used.
Topical cosmetic compositions for use in the invention may also be formulated in a single phase carrier such as water and/or one or more water miscible organic liquids (such as the monohydric and polyhydric alcohols and derivatives thereof described above).
Topical cosmetic compositions for use in the invention may also be formulated in solid forms such as gels or sticks.
Combinations of any of the above described product forms may also be used.
Skin Care Actives
In addition to the galacto-oligosaccharide, a topical cosmetic composition for use in the invention may include additional skin care actives (for improving the physical and/or aesthetic characteristics of the skin). Examples of additional skin care actives which are suitable for use in the invention include vitamins, minerals and/or antioxidants, emollients, humectants, skin anti-hyperpigmentation agents, sunscreens, anti-irritants, exfoliating agents and mixtures thereof.
Vitamins, minerals and/or antioxidants suitable for use in the invention include natural botanical antioxidants derived from plant materials such as fruits, vegetables, herbs and spices (such as goji berry, white tea, rosemary, green tea, grape seed and lemongrass extracts); vitamin A and its precursors or derivatives (such as beta-carotene, retinyl palmitate); vitamin B3 and its precursors or derivatives (such as niacinamide); vitamin B5 and its precursors or derivatives (such as panthenol and its precursors or derivatives); vitamin C and its precursors or derivatives (such as tetrahexyldecyl ascorbate, ascorbyl palmitate); vitamin E and its precursors or derivatives (such as d-alpha-tocopherol, tocopheryl acetate); vitamin K and its precursors or derivatives; selenium and its derivatives (such as L-selenomethionine); and alpha lipoic acid.
Emollients suitable for use in the invention act to increase and maintain moisture in the skin by providing a protective coating to impede epidermal water loss. Examples of emollients include C10-20 fatty alcohols and acids (such as cetyl, myristyl, palmitic and stearyl alcohols and acids); and C10-40 hydrocarbons (such as mineral oil, petroleum jelly, squalene and isoparaffins).
Humectants suitable for use in the invention act to increase and maintain moisture in the skin by attracting water to the stratum corneum of the epidermis. Examples of humectants include amino acids, chondroitin sulfate, glycerin, diglycerin, triglycerin, polyglycerin, polypropylene glycol, polyethylene glycol, ethylene glycol, diethylene glycol, triethylene glycol, propylene glycol, dipropylene glycol, hexylene glycol, 1,3-butylene glycol, 1,4- butylene glycol, ethylene glycol monoalkyl ether, diethylene glycol monoalkyl ether, erythritol, fructose, glucose, maltose, sucrose, lactose, xylose, inositol, lactitol, xylitol, sorbitol, mannitol, maltitol, panthenol, pentaerythritol, 1,2,6-hexanetriol, honey, hyaluronic acid, hydrogenated honey, hydrogenated starch hydrolysate, natural moisturization factor, PEG-15 butanediol, polyglyceryl sorbitol, sodium and potassium salts of pyrollidone carboxylic acid, sodium glucuronate, trehalose and urea.
Skin anti-hyperpigmentation agents suitable for use in the invention include natural botanical agents derived from plant materials (such as Arctostaphylos patula and Arctostaphylos viscida extracts, Emblica officinalis extract, Mitracarpus scaber extract, Uva ursi (bearberry) extract, Morns bombycis (mulberry) extract, Morns alba (white mulberry) extract, Broussonetia papyrifera (paper mulberry) extract, licorice extract, acerola cherry extract, Chlorella vulgaris extract, Aloe ferrox extract and Rumex occidentalis extract); synthetic or natural sugar amines (such as glucosamine, N-acetyl glucosamine, glucosamine sulfate, mannosamine, N-acetyl mannosamine, galactosamine, N-acetyl galactosamine and their hydrochloride salts); retinoids (such as retinol, retinal, retinoic acid and C2-20 esters of retinol such as retinyl palmitate, retinyl acetate, retinyl propionate, retinyl linoleate and retinyl oleate); kojic acid and C2-20 esters thereof (such as kojic acid monobutyrate, kojic acid monocaprate, kojic acid monopalmitate, kojic acid monostearate and kojic acid monobenzoate); hydroquinone, hydroquinone monomethyl ether, hydroquinone monoethyl ether, hydroquinone monobenzyl ether, a and b-arbutin, deoxyarbutin (4-[(tetrahydro-2H- pyran-2-yl)oxy]phenol), mequinol (4-hydroxyanisole), glutathione, cysteine, N-acetyl-L- cysteine, azelaic acid, magnesium ascorbyl phosphate, sodium ascorbyl phosphate, peroxides (such as hydrogen peroxide, zinc peroxide, sodium peroxide and benzoyl peroxide); 3-aminotyrosine, glycyrrhizinic acid and 4-substituted resorcinol derivatives (such as 4-methyl resorcinol, 4-ethyl resorcinol, 4-propyl resorcinol, 4-isopropyl resorcinol, 4-butyl resorcinol, 4-pentyl resorcinol, 4-hexyl resorcinol, 4-heptyl resorcinol, 4-octyl resorcinol, 4- nonyl resorcinol, 4-decyl resorcinol, 4-undecyl resorcinol, 4-dodecyl resorcinol, 4-cyclopentyl resorcinol, 4-cyclohexyl resorcinol, 4-cycloheptyl resorcinol, and 4-cyclooctyl resorcinol).
Sunscreens suitable for use in the invention protect the skin from ultraviolet (UV) solar radiation falling within both the UVB region (between 290nm to 320 nm wavelengths) and the UVA region (between 320nm and 400nm wavelengths). Examples of sunscreens include methoxycinnamate derivatives (such as octyl methoxycinnamate and isoamyl methoxycinnamate); camphor derivatives (such as 4-methyl benzylidene camphor, camphor benzalkonium methosulfate, and terephthalylidene dicamphor sulfonic acid); salicylate derivatives (such as octyl salicylate and homosalate); sulfonic acid derivatives (such as phenylbenzimidazole sulfonic acid); benzone derivatives (such as dioxybenzone, sulisobenzone, and oxybenzone); benzoic acid derivatives (such as aminobenzoic acid and octyldimethyl para-amino benzoic acid); octocrylene, diethylhexyl butamido triazone, octyl triazone, butyl methoxydibenzoyl methane, drometrizole trisiloxane, menthyl anthranilate and inorganic UV-absorbing particles (such as zinc oxide and titanium dioxide). Anti-irritants suitable for use in the invention include allantoin, aloe vera, a-bisabolol, caffeine, chamomile extract, Cola nitada extract, cucumber extract, dipotassium glycyrrhizinate, glycyrrhizic acid, green tea extract, lecithin or hydrogenated lecithin, licorice extract, Avena sativa (oat) meal extract, tea tree oil, salicylic acid, acetylsalicylic acid, strontium acetate, strontium chloride, strontium nitrate, fatty acids with anti-irritant properties (such as linoleic acid and linolenic acid) and aromatic aldehydes with anti-irritant properties (such as 4-methoxy benzaldehyde, 4-ethoxy benzaldehyde, 4-butoxy benzaldehyde and 4- pentoxy benzaldehyde).
Exfoliating agents suitable for use in the invention include benzoyl peroxide, benzoic acid, 3- hydroxy benzoic acid, salicylic acid, acetic acid, trichloroacetic acid, 1-pyrrolidone-5- carboxylic acid, a-hydroxy acids (such as glycolic acid, lactic acid, malic acid, tartaric acid, and citric acid); b-hydroxy acids (such as b-hydroxybutyric acid); a-keto acids (such as pyruvic acid, 2-oxopropanoic acid, 2-oxobutanoic acid and 2-oxopentanoic acid); and oxa acids (such as 3,6,9-trioxaundecanedioic acid).
Mixtures of any of the above described materials may also be used.
Functional Ingredients
In addition to the galacto-oligosaccharide, a topical cosmetic composition for use in the invention may include additional functional ingredients (for improving the physical and/or aesthetic characteristics of the composition).
Examples of additional functional ingredients which are suitable for use in the invention include water soluble or colloidally water soluble polymeric thickening agents (such as hydroxyethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, polyquaternium-10, carrageenan, guar gum, hydroxypropyl guar gum, xanthan gum, polyvinylalcohol, acrylic acid/ethyl acrylate copolymers, carboxyvinyl polymers, cross-linked polyacrylate polymers and polyacrylamide polymers); structurant clays (such as magnesium aluminum silicate, attapulgite, bentonite, montmorillonite and hectorite); inorganic pigments (such as titanium oxide, zirconium oxide, cerium oxide zinc oxide, iron oxide, chromium oxide and ferric blue); organic pigments (such as carbon black and barium, strontium, calcium, and aluminium lakes); pearlescent agents (such as mica coated with titanium oxide and/or iron oxide); dyes, preservatives (such as disodium EDTA, benzyl alcohol, methylparaben, phenoxyethanol, propylparaben, ethylparaben, butylparaben and isobutylparaben); pH adjusters and fragrances.
Mixtures of any of the above described materials may also be used.
Packaging and Use
A topical cosmetic composition for use in the invention (as described above) may be packaged in a suitable container to suit its viscosity and intended use by the consumer. For example, a lotion or a cream can be packaged in a bottle or a roll-ball applicator, or a propellant-driven aerosol device or a container fitted with a pump suitable for finger operation. When the composition is a cream, it can simply be stored in a non-deformable bottle or squeeze container, such as a tube or a lidded jar. The composition is suitably applied to the skin, preferably infant skin, at the rate of one or two applications per day. Generally, an amount corresponding to about 1 to 2 ml of the composition per application is applied uniformly over the area of treatment twice daily for a period of at least 7 (seven) days, more preferably at least 30 (thirty) days. The invention will be further illustrated by the following, non-limiting Examples.
EXAMPLES
Galacto-oligosaccharide (GOS) was evaluated for its ability to enhance growth and acid production of various commensal species of Streptococcus. A stock solution of GOS was prepared such that upon dilution the concentrations in test would range from 2% to 0.07%.
Test bacterial inoculums were prepared in culture media for 18 hours at 37 °C under C02 conditions. Inoculums were centrifuged and resuspended in liquid media diluted to 50% in sterile distilled water. The inoculums were adjusted to a cell density of 1.5-5 x 10s CFU/ml.
To either a 96-well microtiter plate, or a 96-well PreSens HydroPlate®, 90 pi of GOS solution, 90 mI of liquid media at 50% strength and 20 mI of bacterial inoculum was added. Growth controls were included where 90 mI of sterile distilled water, 90 mI 50% liquid media and 20 mI of inoculum were added. For the HydroPlate® pH buffer controls were included.
The microtiter plate was incubated without shaking at 37 °C for 24 hours and an endpoint optical density read taken at 620 nm. The HydroPlate® was read kinetically for fluorescence for 24 hours at 37°C. PreSens calibration software pHSolver_v10 was used as part of the analysis.
The following Table 1 shows the endpoint optical density of the tested bacterial species after incubation with GOS, compared to the growth control.
Table 1
Figure imgf000014_0001
(1) Galacto-oligosaccharide (ex Carbosynth Limited) The following Table 2 shows the calculated acid produced by the tested bacterial species after incubation with GOS, compared to the growth control.
Table 2
Figure imgf000015_0001
The following Table 3 shows the endpoint optical density and calculated acid produced when S. thermophilus (ATCC® 19258™) is incubated with different concentrations of GOS, compared to the growth control.
Table 3
Figure imgf000015_0002
It can be seen from the results that the commensal species of Streptococcus, especially S.thermophilus, show significantly greater growth and acid production when supplemented with GOS in accordance with the invention, as compared to the counterpart control wells containing only dilute liquid media.

Claims

1. The cosmetic use of galacto-oligosaccharides in a topical skin care composition as an active ingredient for providing skin care benefits by supporting the growth of beneficial commensal microbial species which are resident in the human cutaneous microbiome, , characterised in that the beneficial commensal microbial species are Streptococcus species which are a member of the salivarius group of the viridans group of streptococci (VGS); and in that the human is an infant (which is a human baby or young child from birth up to 5 years of age).
2. Galacto-oligosaccharides for use in supporting the growth of beneficial commensal Streptococcus species in the human cutaneous microbiome; characterised in that the beneficial commensal microbial species are Streptococcus species which are a member of the salivarius group of the viridans group of streptococci (VGS); and in that the human is an infant (which is a human baby or young child from birth up to 5 years of age).
3. Use of galacto-oligosaccharides in, and for the manufacture of, topical compositions for supporting the growth of beneficial commensal Streptococcus species which are resident in the human cutaneous microbiome; characterised in that the beneficial commensal microbial species are Streptococcus species which are a member of the salivarius group of the viridans group of streptococci (VGS); and in that the human is an infant (which is a human baby or young child from birth up to 5 years of age).
4. Use according to any one of claims 1 to 3,, in which the beneficial commensal
Streptococcus species is S. thermophilus.
5. Use according to any one of claims 1 to 4, , in which the infant is 1 year old or less.
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US20150202136A1 (en) * 2012-02-14 2015-07-23 The Procter & Gamble Company Topical use of a skin-commensal prebiotic agent and compositions containing the same

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Title
JENNI LEHTIMÄKI ET AL: "Patterns in the skin microbiota differ in children and teenagers between rural and urban environments", SCIENTIFIC REPORTS, vol. 7, 31 March 2017 (2017-03-31), pages 45651, XP055473253, DOI: 10.1038/srep45651 *

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