WO2020052916A1

WO2020052916A1 - A topical composition comprising saccharide isomerate for microbiome balancing

Info

Publication number: WO2020052916A1
Application number: PCT/EP2019/072119
Authority: WO
Inventors: Shanthi APPAVOO; Anindya Dasgupta; Jessica Ann KRISIAK; Srikala KUMARAN; Vivek Subramanian
Original assignee: Unilever N.V.; Unilever Plc; Conopco, Inc., D/B/A Unilever
Priority date: 2018-09-11
Filing date: 2019-08-19
Publication date: 2020-03-19
Also published as: EP3849509A1; CN112672728B; CN112672728A

Abstract

The present invention relates to a topical composition comprising saccharide isomerate for use as a prebiotic when applied on a surface of the human body.

Description

A Topical Composition Comprising Saccharide Isomerate for Microbiome

Balancing

Field of the invention

The present invention relates to a topical composition comprising saccharide isomerate for use as a prebiotic.

Background of the invention

People often try to take care of themselves and of their body surfaces e.g. skin, scalp including hairs, axilla and oral cavity, with a desire of enjoying a healthy lifestyle. Some of the benefits people tend to have desire for include healthy and infection-free skin, even skin tone, adequate moisturization and protection from ultraviolet rays contained in the sunlight. Skin is the outermost protective covering of living beings and is the largest organ in the body. It acts as a barrier and protects the body from external factors. For example, skin prevents entry of harmful or potentially harmful microbes e.g. bacteria, fungi and viruses, from entering the body thereby preventing infection and/or other ill effects that may be caused.

Being the outermost covering however, skin is easily colonized by and constantly remains in contact with microorganisms e.g. bacteria, fungi, viruses, archaea (collectively referred to as microbiome), that may be present on skin. For example, in humans, skin microbiome comprises bacteria that provide a benefit to the host (referred to as commensals or good bugs) e.g.

Staphylococcus epidermidis (S. epidermidis) and; harmful or potentially harmful bacteria (bad bugs) e.g. Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa). Bacteria like S. aureus and P. aeruginosa are not harmful whilst being present on the skin perse. Flowever, upon entry in to the human body through e.g. ingestion and through cuts in the skin, these bacteria may become harmful. Thus, skin microbiome comprises both, good as well as bad bugs. A disruption in the balance between the good and the bad bugs is an opportunity when a disease can set in. Therefore, obtaining and maintaining microbiome balancing, i.e. maintaining a surface of the human body, e.g. skin, in healthy and infection free condition, is often desired by people.

Microbiome balancing may be achieved e.g. by reducing bad bugs through an application of topical compositions comprising one or more antimicrobial compounds. However, most antimicrobial compounds if not all; contained in such topical compositions, do not work in a selective manner. In other words, antimicrobial compounds do not selectively act only against bad bugs but they also act on good bugs. Thus, usage of such topical compositions often leads to depletion in number of good bugs which is not desired.

Certain mechanisms in the human body contribute to microbiome balancing.

For example, keratinocytes, one of the major cell types that is abundantly present in a surface of the human body, e.g. skin, are known to secrete antimicrobial peptides (natural defense system) which primes skin against an attack from bad bugs.

Microbiome balancing may also be achieved by way of utilizing a prebiotic that promotes growth of good bugs over bad bugs. Usage of a prebiotic thus leads to increase in benefits obtained from good bugs. Good bugs are said to provide benefit to their host through e.g. competing with bad bugs for availability of nutrients and through secretion of metabolites.

Several prebiotic ingredients are known in the art. WO2017/201296 (Dermala Inc.) discloses compositions and methods for treating acne vulgaris and cites glycerol, maltose, lactose as prebiotic compounds.

An ingredient‘saccharide isomerate’ a well-known hydrating agent, has been used in compositions. For example, WO 01/70189 (Unilever, 2001 ), US5858997 (Unilever, 1999) and WO 97/47280 (Unilever, 1997) relate to methods and composition for skin lightening that comprises saccharide isomerate. WO 97/30690 (Unilever, 1997) discloses skin treatment composition comprising 1 % saccharide isomerate. US5486352 (Unilever, 1996) discloses sunscreen composition comprising 0.2% saccharide isomerate. EP 0 545 716 (Unilever, 1992) discloses a two-phase clear-opaque toilet bar composition comprising pentavitin (saccharide isomerate commercially available from DSM as a moisturizing agent under trade name Pentavitin^®) wherein it is disclosed as skin treatment active material.

ZA980040 (Elma Mckenzie) discloses a cosmetic skin treatment composition. In a preferred embodiment, ZA980040 discloses the cosmetic skin treatment composition that includes 4% saccharide isomerate acting as the keratin binding complex.

W02017/019612 (Mary Kay Inc.) discloses methods and compositions useful for application to skin and hair comprising hydrolyzed algae extract, saccharide isomerate, and a dermatologically acceptable vehicle wherein the composition or method is capable of moisturizing and/or improving the appearance and/or condition of skin and/or hair.

WO2015/175536 (Biowish Technologies Inc.) provides compositions and methods for improving human health and nutrition. In one aspect it discloses compositions containing a mixture of a prebiotic, a probiotic comprising a mixture of Lactobacillus and /or Bifidobacterium microorganisms produced by solid substrate and submerged liquid fermentation and; a postbiotic derived from the liquid fermentation medium of the Lactobacillus and/or Bifidobacterium microorganisms, collectively referred to as Microbiotic Composites™. It further cites prebiotics e.g. inulin, fructo-oligosaccharides and gluco-oligosaccharide. WO2015175536, in an example therein, describes a deep moisturizer cream that comprises Microbiotic Composites™ along with other ingredients including saccharide isomerate.

It has now been found that a topical composition comprising saccharide isomerate when applied on a surface of the human body, provides microbiome balancing. It has also been found that the microbiome balancing so obtained is by way of saccharide isomerate functioning as a prebiotic, i.e. it preferentially promotes growth of good bugs over bad bags. The benefit of microbiome balancing could take place through various mechanisms and it is not necessary that every substance that provides microbiome balancing does so by way of the substance being a prebiotic. The invention in the present application is by way of a finding that saccharide isomerate behaves as a prebiotic which, to the knowledge of the present inventors, has not been known heretofore.

Summary of the invention

In a first aspect, the present invention relates to use of a topical composition comprising saccharide isomerate as a prebiotic when applied on a surface of the human body.

Detailed description of the invention

Any feature of one aspect of the present invention may be utilized in any other aspect of the invention. The word“comprising” is intended to mean“including” but not necessarily“consisting of or“composed of.” In other words, the listed steps or options need not be exhaustive. Except in the operating and

comparative examples, or where otherwise explicitly indicated, all numbers in this description indicating amounts of material or conditions of reaction, physical properties of materials and/or use are to be understood as modified by the word “about”. Numerical ranges expressed in the format "x to y" are understood to include x and y. When for a specific feature multiple preferred ranges are described in the format "x to y", it is understood that all ranges combining the different endpoints are also contemplated. Unless specified otherwise, amounts as used herein are expressed in percentage by weight based on total weight of the composition and is abbreviated as“wt%”. The use of any and all examples or exemplary language e.g.“such as” provided herein is intended merely to better illuminate the invention and does not in any way limit the scope of the invention otherwise claimed.

The phrase“microbiome balancing” used herein preferably means maintaining a surface of the human body e.g. skin, scalp including hairs, axilla and oral cavity, in healthy and infection free condition. Microbiome balancing is obtained by way of saccharide isomerate functioning as a prebiotic, i.e. preferentially promoting growth of good bugs over bad bugs, on a surface of the human body.

In a first aspect, the present invention relates to a topical composition

comprising saccharide isomerate for use in providing microbiome balancing when applied on a surface of the human body.

It has been found that a topical composition according to the present invention (the composition) comprising saccharide isomerate provides microbiome balancing.

Microbiome balancing so obtained is through saccharide isomerate functioning as a prebiotic. Saccharide isomerate is found to preferentially promote growth of good bugs, e.g. S. epidermidis over growth of bad bugs e.g. S. aureus. Thus, it has also been found that saccharide isomerate functions as a prebiotic.

Preferably, the composition comprises saccharide isomerate in amounts from 0.001 to 10 wt%, more preferably from 0.01 to 8 wt%, even more preferably from 0.1 to 7 wt%, further more preferably from 0.1 to 5 wt% and still more preferably from 0.1 to 3 wt%. Saccharide isomerate is commercially available under trade name‘Pentavitin^®’ (from DSM), under trade name Waterin (from Clariant), under trade names ‘EPS3 Powder’,‘EPS4 Powder’,‘EPS5 Powder’ and‘EPS15 Powder’ (all from Codif) and under trade name‘Hyanify’ (from Lipotec S.A.).

Preferably, the composition further comprises an additional prebiotic selected from polyols, monosaccharides, disaccharides, oligosaccharides,

polysaccharides and mixtures thereof. Examples of polyols that may be used as an additional prebiotic include glycerol, sugar alcohols e.g. maltitol, sorbitol, xylitol, erythritol and isomalt and mixtures thereof.

Examples of monosaccharide that may be used as an additional prebiotic include glucose, fructose, galactose and mixtures thereof.

Examples of disaccharides that may be used as an additional prebiotic include sucrose, maltose, lactose and mixtures thereof. Examples of oligosaccharides that may be used as an additional prebiotic include fructo-oligosaccharides, gluco-oligosaccharide and mixtures thereof. Examples of polysaccharides that may be used as an additional prebiotic include inulin. When present, an additional prebiotic may be present in the composition preferably from 0.001 to 10 wt%, more preferably from 0.01 to 8 wt%, even more preferably from 0.1 to 7 wt%, further more preferably from 0.1 to 5 wt% and still more preferably from 0.1 to 3 wt%. When an additional prebiotic may be present in the composition, it may be present in a ratio preferably from 20:1 to 1 :20 to saccharide isomerate, more preferably from 10:1 to 1 :10, even more preferably from 5:1 to 1 :5, further more preferably from 2:1 to 1 :2 and still more preferably in ratio 1 :1 to saccharide isomerate.

A topical composition according to the first aspect provides microbiome balancing when applied on a surface of the human body. Preferably, a surface of the human body is at least one of skin, scalp including hairs, axilla and oral cavity.

In a second aspect, the present invention relates to use of a topical composition comprising saccharide isomerate for microbiome balancing when applied on a surface of the human body. Microbiome balancing is obtained by way of use of a topical composition comprising saccharide isomerate on a surface of the human body. Preferably, a surface of the human body is at least one of skin, scalp including hairs, axilla and oral cavity. The use according to this aspect can be therapeutic or non-therapeutic. Preferably, the use is non-therapeutic, including cosmetic.

In a third aspect, the present invention relates to use of a topical composition comprising saccharide isomerate as a prebiotic. When a topical composition comprising saccharide isomerate is applied on a surface of the human body, it is found to preferentially promote growth of good bugs e.g. S. epidermidis over growth of bad bugs e.g. S. aureus. Preferably, a surface of the human body is at least one of skin, scalp including hairs, axilla and oral cavity. The use according to this aspect can be therapeutic or non-therapeutic. Preferably, the use is non-therapeutic, including cosmetic.

Because of increase in number of good bugs upon exposure to saccharide isomerate, metabolites secreted by good bugs are also likely to increase.

Without wishing to be bound by theory, these metabolites are believed to contribute to the natural-defense system of the human body. Accordingly, in a fourth aspect, the present invention relates to use of topical a composition comprising saccharide isomerate for improving natural -defense system of a surface of the human body by way of microbiome balancing.

Preferably, a surface of the human body is at least one of skin, scalp including hairs, axilla and oral cavity. The use according to this aspect can be therapeutic or non-therapeutic. Preferably, the use is non-therapeutic, including cosmetic.

Accordingly, in a fifth aspect, the present invention relates to a method of improving natural-defense system of a surface of the human body by way of microbiome balancing comprising the step of applying the composition according to the first aspect to the surface of the human body. Preferably, a surface of the human body is at least one of skin, scalp including hairs, axilla and oral cavity. The method according to this aspect can be therapeutic or non- therapeutic. Preferably, the method is non-therapeutic, including cosmetic.

Preferably, the composition comprises a cosmetically acceptable base.

Examples of ingredients that may be used as cosmetically acceptable base includes water, fatty acids, soaps (salts of fatty acids), alcohols and mixtures thereof.

Preferably, the composition comprises water in an amount from 1 to 99 wt%, more preferably from 5 to 90 wt%, even more preferably from 35 to 80 wt% and further more preferably from 40 to 75 wt%. Examples of alcohols that may be used as cosmetically acceptable base include the alcohol having straight or branched chain of carbon atoms preferably containing from 1 to 16, more preferably from 2 to 10, even more preferably from 3 to 8 carbon atoms. Illustrative examples of alcohol that may be used in the composition include methyl alcohol, ethyl alcohol, isopropyl alcohol, n-propyl alcohol, isobutyl alcohol, n-butyl alcohol, n-pentyl alcohol, n- hexyl alcohol and mixtures thereof. Preferably, alcohol is selected from ethyl alcohol, isopropyl alcohol and mixtures thereof.

Preferably, the composition comprises alcohols in amount ranging preferably from 0.1 to 80 wt%, more preferably from 0.1 to 65 wt%, even more preferably from 1 to 50 wt% and further more preferably from 5 to 30 wt%.

Examples of fatty acids that may be used as cosmetically acceptable base include fatty acids that have from 12 to 20 carbon atoms, more preferably from 14 to 18 carbon atoms. Examples of such fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid and mixtures thereof. The most preferred fatty acid is stearic acid. The fatty acid may also be hystric acid which is substantially (generally about 90 to 95%) a mixture of stearic acid and palmitic acid in a ratio of between 55:45 to 45:55.

Preferably, cosmetically acceptable base comprises from 1 to 25 wt%, more preferably from 3 to 22 wt%, even more preferably from 5 to 20 wt% and still more preferably from 6 to 18 wt% fatty acid. Soap may be formed in-situ by addition of alkaline agents e.g. sodium

hydroxide or potassium hydroxide; or may be added externally to the

composition. For example, when potassium hydroxide is added to cosmetically acceptable base comprising stearic acid, potassium stearate thus formed, is understood to be the soap formed in-situ which forms a part of cosmetically acceptable base.

Preferably, the composition comprises from 0.1 to 10 wt%, more preferably from 0.1 to 7 wt% even more preferably from 0.1 to 3 wt% soap. The composition is preferably in the form of a wash-off or a leave-on

composition, more preferably a leave-on composition. Leave-on composition preferably means a composition which is not required to be removed from the human body after the application of said composition. When the composition is in the form of a leave-on composition, the composition may be in the form of a deodorant (stick, roll-on or spray), a hand sanitizer, a body lotion, a skin cream and a body spray.

For example, if the composition is in the form of a skin cream, a vanishing cream base that generally comprises from 5 to 25 wt% fatty acid and from 0.1 to 10 wt% soap and from 60 to 80 wt% water as the cosmetically acceptable base, is preferred. Vanishing cream base gives a highly appreciated matt feel to the skin.

Additionally, if the composition is in the form of a deodorant, the composition may preferably comprise a conventional deodorant base as the cosmetically acceptable base. By a deodorant is meant a product in the stick, roll-on, or propellant medium which is used for personal deodorant benefit e.g. application in axilla, the under-arm area, which may or may not contain anti-perspirant actives.

Deodorant compositions can generally be in the form of firm solids, soft solids, gels, creams, and liquids and are dispensed using applicators appropriate to the physical characteristics of the composition. Deodorant compositions which are delivered through roll-ons generally comprise a liquid carrier. Such liquid carrier can be hydrophobic or comprise a mixture of both hydrophilic and hydrophobic liquids. They may be in the form of an emulsion or a microemulsion. The liquid carrier or mixture of carriers often constitutes from 30 to 95 wt% and in many instances from 40 to 80 wt%. Hydrophobic liquid carriers commonly can comprise one or more materials selected within the chemical classes of siloxanes, hydrocarbons, branched aliphatic alcohols, esters and ethers that have a melting point not higher than 25°C and a boiling point of at least 100°C. Hydrophilic carrier liquids that can be employed in compositions herein commonly comprise water and/or a mono or polyhydric alcohol or water- miscible homologue. Polyhydric alcohols commonly comprise ethylene or propylene glycol, or a homologue can be employed such as diethylene glycol. Other than this suitable other vehicle and component used for deodorant composition can be added.

Wash-off composition preferably means a composition which is

intended/required to be removed from the body by washing with solvent preferably water after the application of said composition like e.g. pastes including a toothpaste, a shampoo, a hand wash composition and a face wash composition. In case of wash-off compositions, a cosmetically acceptable base preferably further comprises a surfactant.

For example, if the composition is in the form of a shampoo, in addition to water, the cosmetically acceptable base preferably comprises an anionic surfactant e.g. an alkyl sulphate and/or ethoxylated alkyl sulfate surfactant. These anionic surfactants are preferably present at a level of from 1 to 20 wt%, more preferably from 2 to 16 wt%, even more preferably from 3 to 16 wt%.

Preferred alkyl sulfates are C8-18 alky sulfates, more preferably C12-18 alkyl sulfates, preferably in the form of a salt with a solubilizing cation such as sodium, potassium, ammonium or substituted ammonium.

Preferably, the composition further comprises one or more skin lightening agents. These one or more skin lightening agents may be selected from niacinamide, resorcinol, phenylethyl resorcinol, 4-alkyl substituted resorcinol e.g. 4-ethyl resorcinol and 4-hexyl resorcinol, vitamin B6, vitamin C, vitamin A, glutathione precursors, galardin, adapalene, aloe extract, ammonium lactate, arbutin, azelaic acid, butyl hydroxy anisole, butyl hydroxy toluene, citrate esters, deoxyarbutin, 1 ,3-diphenyl propane derivatives, 2,5-dihydroxybenzoic acid and its derivatives, 2-(4-acetoxyphenyl)-1 ,3-dithiane, 2-(4-hydroxyphenyl)-1 ,3- dithiane, ellagic acid, gluco pyranosyl-1 -ascorbate, gluconic acid, glycolic acid, green tea extract, 4-Hydroxy-5-methyl-3[2H]-furanone, 4-hydroxyanisole and its derivatives, 4-hydroxybenzoic acid derivatives, hydroxycaprylic acid, inositol ascorbate, lactic acid, lemon extract, linoleic acid, magnesium ascorbyl phosphate, 5-octanoyl salicylic acid, salicylic acid, 3,4,5-trihydroxybenzyl derivatives, acetylglucosamine, pitera extract, symwhite, calcium pantothenate (Melano-block), seppiwhite, soybean extract (bowman birk inhibitor), 12- hydroxystearic acid and mixtures thereof. When used in the composition, 12- hydroxystearic acid is used as a skin lightening agent and not as a fatty acid. Preferably, skin lightening agents that may be used in the composition are niacinamide, resorcinol, phenylethyl resorcinol, 4-alkyl substituted resorcinol e.g. 4-ethyl resorcinol, 4- isopropyl resorcinol and 4-hexyl resorcinol, vitamin B6, 12-hydroxystearic acid, glutathione precursors, galardin and mixtures thereof.

When incorporated, a skin lightening agent may be added in an amount preferably from 0.001 to 15 wt%, more preferably from 0.01 to 10 wt%, even more preferably from 0.1 to 5 wt% and further more preferably from 0.5 to 3 wt%, in the composition.

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

When present in the composition, preservatives are added preferably in an amount from 0.001 to 5 wt%, more preferably from 0.01 to 3 wt% and most preferably from 0.02 to 2 wt%.

Preferably, the composition further comprises a range of other optional ingredients that include antioxidants, binders, biological additives, buffering agents, colorants, astringents, fragrance, opacifying agents, conditioners, exfoliating agents, pH adjusters, natural extracts, skin sensates, skin soothing agents, emollients, solvents and skin healing agents.

Preferably, the composition is formulated in the form of a powder, flake, lotion, cream, gel or mousse. More preferably, the composition is formulated in the form of cream or lotion and most preferably in the form of cream. The

composition is preferably of leave-on type. The packaging for the composition of this invention can be a patch, bottle, tube, roll-ball applicator, propellant driven aerosol device, squeeze container or lidded jar.

The present invention now will be demonstrated by way of following non-limiting examples.

Examples

Protocols

Preparation of cultures

Cultures of bacteria S. epidermidis (ATCC 12228), S. aureus (ATCC 6538) and P. aeruginosa (ATCC 15442) were maintained in Tryptic soy broth (TSB; Difco catalogue number 21 1825) at 10⁸ log colony forming units (cfu)/ml_ and used in the experiments at appropriate dilutions specified below. Example 1 : Saccharide Isomerate preferentially promotes growth of a good bug e.q. S. epidermidis over a bad bug e.q. S. aureus.

S. aureus and S. epidermidis were grown in TSB in presence or absence of 2% saccharide isomerate for 24 h at 37°C. At the end of incubation, optical density at 620 nm was recorded for all the test samples. The ratio of growth of the bacteria in presence and absence of saccharide isomerate was found to be as given below:

The data in table above shows that saccharide isomerate preferentially supports growth of S. epidermidis over S. aureus. Saccharide isomerate thus functions as a prebiotic thereby providing microbiome balancing.

Example 2A, 2B and 2C: Growth Inhibition study

Overnight grown plate culture of S. epidermidis was serially diluted in sterile saline to 10⁶ log cfu/mL. 100 pl_ of this solution was added to 40 ml_ of TSB in absence or presence of 1 % saccharide isomerate and incubated aerobically under shaking condition at 37°C for 24 h. At the end of 24 h, the cultures were pelleted down by centrifugation at 8000 rpm for 2 mins at room temperature and ferments (the supernatant) were collected. The ferment A (collected from where saccharide isomerate was absent) and ferment B (where 1 % saccharide isomerate was present), were filter sterilized with 0.22 pm filters. The ferments were checked for pH and used in growth inhibition study. The study was carried out in a 96-well plate using test conditions outlined in examples 2A, 2B and 2C below. Example 2A

The following test samples were prepared in a 96-well plate and the plate was incubated at 37°C in a plate reader for 24 h.

- 100 pl_ ferment A + 100 pl_ fresh TSB + 20 pl_ of S. epidermidis (10⁶ Log cfu/mL); and

- 100 pL ferment B + 100 pL fresh TSB + 20 pL of S. epidermidis (10⁶ Log cfu/mL)

The optical density was recorded at an interval of every 1 h and; was found to be as shown in table 2A below:

Table 2A: S. epidermidis ferment against S. epidermidis

The data in table 2A above shows that neither ferment A (obtained from of S. epidermidis in absence of saccharide isomerate) nor ferment B (obtained from of S. epidermidis in presence of 1 % saccharide isomerate) inhibit growth of S. epidermidis itself.

Example 2B

Further, the following test samples were prepared in a 96-well plate and the plate was incubated at 37°C in a plate reader for 24 h.

- 100 pl_ ferment A + 100 mI_ fresh TSB + 20 mI_ of S. aureus (10⁶ Log cfu/mL); and

- 100 pL ferment B + 100 pL fresh TSB + 20 pL of S. aureus (10⁶ Log cfu/mL)

The optical density was recorded at an interval of every 1 h and; was found to be as shown in table 2B below:

Table 2B: S. epidermidis ferment against S. aureus

The data in table 2B above shows that the ferment B (obtained from S.

epidermidis in presence of 1 % saccharide isomerate) inhibits growth of S. aureus as compared to ferment A (obtained from S. epidermidis in absence of saccharide isomerate).

Example 2C

- 100 pl_ ferment A + 100 pl_ fresh TSB + 20 pl_ of P. aeruginosa (10⁶ Log cfu/mL); and

- 100 pL ferment B + 100 pL fresh TSB + 20 pL of P. aeruginosa (10⁶ Log cfu/mL). Table 2C: S. epidermidis ferment against P. aeruginosa

The data in table 2C above shows that ferment B (obtained from S. epidermidis in presence of 1 % saccharide isomerate) inhibits growth of P. aeruginosa as compared to ferment A (obtained from S. epidermidis in absence of saccharide isomerate).

The data in tables 2A, 2B and 2C together indicate that ferment of S.

epidermidis obtained upon fermentation of saccharide isomerate comprises one or more metabolites secreted by S. epidermidis that leads to inhibition of harmful/ potentially harmful bacteria e.g. S. aureus and P. aeruginosa but not S. epidermidis (a commensal) itself.

Example 3: Identification of metabolite produced in fermentation of saccharide isomerate by S. epidermidis.

Metabolites produced in fermentation of saccharide isomerate by S. epidermidis were identified using gas chromatography.

Briefly, for sample preparation, 5 ml_ of the supernatant was taken in a petri plate and dried at 40°C in an oven. After drying, the residue was re-dissolved in 1 N HCI-methanol mixture, followed by addition of 0.5 ml_ of BF3 methanol reagent (methylation reagent) and heated at 75°C for 2 h. The methylated organic acids were analyzed using GC-MS with SIM mode. The compounds were identified using the spectra obtained with standards and cross-verified using the N.I.S.T. library. The retention times and quantification ions of each compound are given in the table.

Quantification of the fatty acids as well as organic acid methyl ester was done using pure standards through external calibration method.

Method for detection: Gas Chromatography (Perkin Elmer Clarus 500 with GCMS Detector) was used. A polar GC column CP WAX 52 CB (30 m X 0.25 mm x 0.25 microns) was selected for the analysis. The injection port

temperature was maintained at 220°C. The desorption time was 3 min. The oven temperature was programmed with an initial temperature of 40°C hold for 2 min; and raised the temperature to 250°C at the ramp of 10°C per minute which was held for 10 min. GCMS was used for the analysis of the actives and the compounds were identified using NIST library match. The mass source temperature was maintained at 200°C with a scan in the range 40 to 500 m/z. Helium was used as a carrier gas and a split less mode of injection was used for this study.

The major metabolite that was produced in fermentation of saccharide isomerate by S. epidermidis was identified to be lactic acid.

Example 4: Dose dependent increase in lactic production in fermentation of saccharide isomerate.

TSB in absence or presence of saccharide isomerate in amounts shown in table below, was incubated at 37°C for 24 h under aerobic conditions. At the end of incubation, the test samples were assayed for lactic acid content using gas chromatography as described above.

The data in table above shows that with increasing amount of saccharide isomerate, increasing amount of lactic acid was produced by S. epidermidis.

Example 5: In vivo study to demonstrate lactic acid production upon

fermentation of saccharide isomerate.

In a 2 day in-vivo study conducted (5 volunteers), 0.5% and 1 % saccharide isomerate prepared in water, was applied 3 times each day on volar fore arm of volunteers. Samples collected (using isopropanol) from volar fore arms on the third day were assayed (using gas chromatography as described above) for fold increase in lactic acid amounts over and above the baseline (no treatment). The results obtained were as in the table below.

The data above shows that upon application of saccharide isomerate on a surface of the human body, here; on the skin, a proportionately increasing amount of lactic acid is detected in-vivo. This indicates that skin microbiome can ferment saccharide isomerate and that, lactic acid is produced in fermentation of saccharide isomerate by bugs present in skin microbiome.

Example 6: Hand sanitizer composition comprising saccharide isomerate

Example 7: A vanishing cream composition comprising saccharide isomerate

In conclusion, from the examples and description above, it is evident that a topical composition comprising saccharide isomerate provides microbiome balancing when applied on a surface of the human body. The microbiome balancing so obtained is found to be by way of saccharide isomerate functioning as a prebiotic. Saccharide isomerate preferentially promoted growth of commensal bacteria e.g. S. epidermidis over harmful/potentially harmful bacteria e.g. S. aureus. The major metabolite in fermentation of saccharide isomerate is found to be lactic acid that contributes to natural defence system of a surface of the human body e.g. skin, scalp including hairs, axilla and oral cavity.

Claims

1. Use of a topical composition comprising saccharide isomerate as a

prebiotic when applied on a surface of the human body.

2. Use according to claim 1 wherein the composition comprises from 0.001 to 10 wt% saccharide isomerate.

3. Use according to claims 1 or 2 wherein the composition further

comprises from 0.001 to 10 wt% of an additional prebiotic.

4. Use according to claim 3 wherein the additional prebiotic is selected from polyols, monosaccharides, disaccharides, oligosaccharides,

polysaccharides and mixtures thereof

5. Use as claimed in any one of the preceding claims wherein the surface of the human body is at least one of skin, scalp including hairs, axilla and oral cavity.

6. Use as claimed in any one of the preceding claims wherein the use is non-therapeutic.