This invention relates to a topical formulation comprising a prebiotic oligosaccharide selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, or combinations thereof, and xylitol, and its use in therapy by topical application.

BACKGROUND OF THE INVENTION

The largest organ of human body is the skin, which plays a pivotal role in protecting the host from pathogenic infections and penetration of harmful agents. Before birth, the skin is completely sterile but after birth, environmental microbes that are in homeostasis with the host colonize it. Moreover, after a vaginal delivery, fecal and vaginal microbes belonging to the mother's bacterial microflora also colonize the skin of infants. The microbial community living on the human skin is called skin microbiota, and it is constituted by over 100 distinct species of bacteria. The skin is the body's interface to the outside world, and it harbors populations of non-pathogenic, commensal microorganisms have an important function in skin health and disease.

Our skin hosts about 10 ⁶ bacteria per square centimeter which form our skin microflora. A healthy balanced skin biota is the microbial shield against pathogenic microorganisms and it can prevent dry skin conditions and improve skin health.

Alterations in the skin microbiome are an important component of the pathophysiology of atopic dermatitis. Skin affected by atopic dermatitis is often colonized by Staphylococcus aureus (S. aureus), particularly during a disease flare, whereas the abundance of other commensal bacteria is markedly decreased. These changes in the skin microbiome negatively affect the epidermal barrier and initiate inflammation. When bacteria such as S. aureus attach to the skin surface, they form biofilms which act as a scaffold for further bacterial growth. Antibiotics are not the optimal treatment for removing bacteria from the skin of atopic dermatitis patients due to issues such as skin irritation and bacterial resistance.

It is an object of the present invention to provide a topical formulation for treating a skin disease or disorder, for example atopic dermatitis.

SUMMARY OF THE INVENTION

In a first aspect, the invention provides a topical formulation comprising:

• a prebiotic oligosaccharide selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, or combinations thereof;

• 0.01 % to 10% w/w xylitol; and

• a pharmaceutically or cosmetically acceptable carrier.

In a second aspect, the invention provides the formulation as defined by the first aspect of the invention for use in therapy.

In a third aspect, the invention provides a formulation comprising: • a prebiotic oligosaccharide selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, or combinations thereof;

• xylitol; and

• a pharmaceutically or cosmetically acceptable carrier for use in therapy by topical application.

In a fourth aspect, the invention provides the formulation as defined by the first aspect of the invention for use as a cosmetic.

Surprisingly, the applicant has found that a formulation comprising a prebiotic oligosaccharide selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, and xylitol, has a beneficial effect on the skin microbiome and thereby is suitable fortreating diseases and disorders of the skin and for cosmetic uses to improve the appearance of the skin.

In a fifth aspect, the invention provides a topical formulation comprising a prebiotic oligosaccharide selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin or dextran, or combinations thereof, for use in treating atopic dermatitis by topical application.

Additional aspects of the invention are more fully described in the following detailed description of the various embodiments.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides a topical formulation comprising:

• a prebiotic oligosaccharide selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, or combinations thereof;

• 0.01% to 10% xylitol; and

• a pharmaceutically or cosmetically acceptable carrier.

Oligosaccharides

Oligosaccharides are saccharide polymers containing fewer monosaccharide units than polysaccharides, but more than the two or three units in di- or tri-saccharides. One exception to this is lactulose, which is a di-saccharide with properties similar to oligosaccharides, and so is considered to form part of the oligosaccharide class (Carbohydrate Polymers 68 (2007) 587-597).

Galatcooligosaccharides (GOS), lactulose, polydextrose, inulin, and fructooligosaccharides (FOS) are oligosaccharides that are established prebiotics. Isomaltooligosaccharides (IMO), arabinogalactan (LAG) and dextran are oligosaccharides that are recognised as emerging prebiotics (Curr Osteoporos Rep, 2015, 13: 363-371). These compounds are non-digestible by humans and promote the growth of beneficial microorganisms, which have the potential to improve host health.

The topical formulations of the present invention comprise fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, or combinations thereof. For the avoidance of doubt, the terms fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran include one or more fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran respectively. Compositions comprising mixtures of one type of oligosaccharides, and compositions comprising mixtures of different types of the prebiotic oligosaccharides, are included in the invention.

Fructooligosaccharides are inulin-type oligosaccharides of D-fructose, joined by a b-(2 1) linkage, with a terminal D-glucosyl residue (Enzyme Microb. Technol., 1996, vol. 19, August 1). FOS are derived either from the hydrolysis of inulin or from sucrose. FOS derived from sucrose are shorter chain oligomers; usually, production from sucrose results in the production of three oligosaccharides; 1- kestose, 1-nystose and 1F-p-fructofuranosylnystose (Annu. Rev. Food Sci. Technol. 2010. 1 :305-39).

In one embodiment, the prebiotic oligosaccharide composition of the present invention is a blend comprising FOS with a degree of polymerisation (DP) of between 2 and 20. In another embodiment, the prebiotic oligosaccharide is a blend comprising FOS with a DP of between 2 and 10, or between 2 and 6. In a further embodiment, the prebiotic oligosaccharide of the instant formulation is a blend comprising FOS with a DP of 2 or more, of 4 or more, of 8 or more, or of 10 or more.

Isomaltooligosaccharides (IMO) are glucose oligomers with a-1 6-glucosidic-linkages. The general term IMO includes, inter alia, isomaltose, panose, isomaltotriose, isomaltotetraose, isomaltopentaose, nigerose, kojibiose and some higher branched oligosaccharides. IMOs can be produced industrially from starch, using an enzyme catalysed process wherein the starch is first liquefied by a-amylase, and subsequently hydrolysed to maltose using b-amalyse. Finally, IMO is produced using a-Glucosidase (Carbohydrate Polymers 68 (2007) 587-597). Honey, miso, sake and soy source are natural sources of IMO.

In one embodiment, the prebiotic oligosaccharide of the present invention is a blend comprising IMO with a DP of between 1 and 20. In another embodiment, the prebiotic oligosaccharide is a blend comprising IMO with a DP of between 2 and 10, or between 2 and 6, or between 1 and 3. In a further embodiment, prebiotic oligosaccharide is a blend comprising IMO with a DP of 6 or more, or of 10 or more, or of 20 or more.

Arabinogalactan comprises a galactan chain, with alternating 5-linked b-D-galactofuranosyl (Gal/) and 6-linked b-D-Galf residues, wherein arabinan chains are joined to the galactan structure at residues 8, 10 and 12. The arabinan chains have three structural domains; a linear 5-linked a-D-Araf residue, a 3, 5-linked a- D-Araf residue substituted with 5-linked a-D-Arafat the branched positions, and finally the non-reducing terminus of arabinan consists of a 3, 5-linked a-D-Araf residue with the b-0-AG8/ ^: -(1-2)- a- D-Araf disaccharide (Bacterial Polysaccharides: current innovations and future trends, 2009). Arabinogalactan is a major component in many gums, and is found naturally in plants such as larch trees.

The DP of arabinogalactan varies with the oligosaccharaide source. Arabinogalactan sourced from white spruce was found to have a DP of 100, whereas the arabinogalactan sources from Douglas fir has been found to have a DP of 340 (Advances in Carbohydrate Chemistry, volume 20, page 432).

Inulin is comprised of polysaccharides containing a backbone of fructose units, usually with a terminal glucose unit. The fructose units are joined by b-(2 1)D-fructosyl-fructose bonds, and the terminal glucose unit joined by an a-D-glucopyranosoyl bond. Occasionally, inulin does not contain any glucose units. The general formula can be shown as GF _n and F _m , wherein G is a terminal glucose monomer, F is the fructose unit and n or m represents the number of fructose monomer units (Journal of Thermal Analysis and Calorimetry, Vol. 71 (2003) 215-224).

Inulin occurs naturally in several plant species, including Liliales, i.e. garlic, asparagus, leek and onion, and Compositae, i.e. Jerusalem artichoke and chicory. Chicory is the predominant commercial source of food grade inulin (Annu. Rev. Food Sci. Technol. 2010. 1 :305-39). The DP varies with the origin of the inulin, generally from 2 to 10, but up to 40 or more in some cases (Food Chemistry 113 (2009) 1058-1065). Inulin derived from chicory has an average DP (DP _av ) of 12, whereas inulin derived from artichoke has a DP _av of 46. In one embodiment, the prebiotic oligosaccharide composition of the present invention is a blend comprising inulin with a DP of from 2 to 20. In a further embodiment, the prebiotic oligosaccharide composition comprises inulin with a DP range of from 5 to 15 or a DP range of from 10 to 15. In a further embodiment, the prebiotic oligosaccharide composition is a blend of one or more saccharides with a DP of 10 or more, 20 or more, of 30 or more or of 40 or more.

Dextran is a homopolysaccharide, with a backbone of composed of a-(1 ,6)-linked d-glucopyranose. Dextran includes a family of polysaccharides with various degrees of branching, involving a-(1 ,2), a- (1 ,3) and a-(1 ,4) linkages. Dextrans are produced by an enzyme catalysed reaction in bacterial sources; the particular bacteria and the type of dextransucrase that catalyses the reaction influences the structure of the resulting dextran by influencing the type and degree of branching in the polysaccharide (Carbohydrate Polymers 88 (2012) 1440-1444). In one embodiment, the prebiotic oligosaccharide composition of the present invention is a blend comprising dextran with a DP of from 2 to 10. In a further embodiment, the prebiotic oligosaccharide composition is a blend comprising dextran with a DP of from 2 to 7, or from 2 to 5, or from 3 to 5.

In one embodiment, the topical formulation comprises a prebiotic agent selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, or combinations thereof.

In a preferred embodiment, the prebiotic agent is selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan and inulin.

In another preferred embodiment, the prebiotic agent is selected from arabinogalactan and isomaltooligosaccharides.

In the most preferred embodiment, the prebiotic agent is selected from fructooligosaccharides.

In one embodiment, the topical formulation comprises 0.01% to 10% w/w of the prebiotic oligosaccharide defined above, for example 0.01% to 5% w/w of the prebiotic oligosaccharide. Alternatively, the formulation comprises 0.1% to 5% w/w of the prebiotic oligosaccharide, for example 0.5% to 5% w/w of the prebiotic oligosaccharide.

In a preferred embodiment, the topical formulation comprises 0.01% to 5% w/w of the prebiotic oligosaccharide. The topical formulation includes 0.01% to 10% w/w xylitol. Xylitol is a naturally occurring sugar alcohol used as a sweetener. It is found in low concentrations in the fibers of many fruits and vegetables and can be extracted from various berries, oats, and mushrooms, as well as fibrous material such as cornhusks and sugar cane bagasse.

In one embodiment, the topical formulation comprises 0.01% to 5% w/w xylitol. Alternatively, the formulation comprises 0.1 % to 5% w/w xylitol or 0.5% to 5% w/w xylitol.

In a preferred embodiment, the topical formulation comprises 0.01% to 5% w/w xylitol.

In one embodiment, the topical formulation comprises 0.01% to 8% w/w xylitol. Alternatively, the formulation comprises 0.1 % to 7% w/w xylitol or 0.5% to 7% w/w xylitol. In a preferred embodiment, the topical formulation comprises 1 % to 7% w/w xylitol, 3% to 6% w/w xylitol, or 4 to 6% w/w xylitol.

In one embodiment, the topical formulation comprises a weight ratio of the prebiotic oligosaccharide:xylitol of from 10:1 to 1 :10, from 8:1 to 1 :8, from 5:1 to 1 :5, or from 3:1 to 1 :3.

In a preferred embodiment, the topical formulation comprises a weight ratio of prebiotic oligosaccharide:xylitol of from 2:1 to 1 :2.

In a preferred embodiment, the topical formulation comprises a weight ratio of prebiotic oligosaccharide:xylitol of from 2:1 to 1 :2.

In another preferred embodiment, the topical formulation comprises a weight ratio of the prebiotic oligosaccharide:xylitol of from 1 :1 to 1 :20, from 1 :5 to 1 :15, or from 1 :7 to 1 :12. The topical formulations are for use in therapy, in particular by topical application.

In one embodiment, the invention provides a topical formulation comprising:

• at least one prebiotic oligosaccharide selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, or combinations thereof;

• xylitol; and

• a pharmaceutically or cosmetically acceptable carrier; for use in therapy by topical application.

In a further embodiment, the invention provides a topical formulation comprising a prebiotic oligosaccharide selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, or combinations thereof, for use in treating atopic dermatitis by topical application. In one embodiment, the topical formulation is for use in the treatment of a skin disease or disorder.

In a further embodiment, the invention provides a topical formulation comprising xylitol for use in therapy, wherein the topical formulation further comprises a prebiotic oligosaccharide selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, or combinations thereof. In another embodiment, the invention provides a topical formulation comprising xylitol for use in the treatment of a skin disease or disorder, wherein the topical formulation further comprises a prebiotic oligosaccharide selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, or combinations thereof. In a further embodiment, the skin disease or disorder is associated with S. aureus. In a still further embodiment, the skin disease or disorder associated with S. aureus is atopic dermatitis.

In one embodiment, the invention provides a method of treating a skin disease or disorder comprising the topical application of an effective amount of topical formulation according to the invention to a patient in need thereof. In another embodiment, the invention provides the use of a prebiotic oligosaccharide selected from fructooligosaccharides, isomaltooligosaccharides, arabinogalactan, inulin and dextran, or combinations thereof, in combination with xylitol for the manufacture of a medicament for use in a method of treating a skin disease or disorder.

In one embodiment, the topical formulations are for use in the treatment of a skin disease or disorder associated with Staphylococcus aureus (S. aureus). A skin disease or disorder associated with S. aureus is one which is caused or promoted by S. aureus.

S. aureus is a gram-positive pathogenic strain responsible for a wide variety of conditions, ranging from clinical inflammation to severe infections causing pneumonia, endocarditis and septicaemia. S. aureus is one of the most important bacteria in human diseases. S. aureus is particularly associated with atopic dermatitis, a common chronic inflammatory skin disease characterized by acute eczematous lesions erythema. Pruritus is also prominent and consistent and thus has an effect on quality of life. Atopic dermatitis affects 10-30% of children and 2-3% of adults in industrialized countries. S. aureus is also a causative or compounding factor in the pathogenesis of other minor skin infections such as pimples, impetigo, cellulitis and folliculitis, and in more serious and invasive diseases such as pneumonia, vaginitis, meningitis, osteomyelitis, toxic shock syndrome, bacteraemia, and sepsis.

In one embodiment, the skin disease or disorder is atopic dermatitis (i.e. atopic eczema), and in particular is atopic dermatitis associated with S. aureus.

Without being bound by theory, it is thought that the topical formulations of the invention promote growth of Staphylococcus epidermidis (S. epidermidis), the most commonly isolated bacteria species in healthy skin which makes up to 90 % of the aerobic flora. S. epidermidis balances the inflammatory response after skin injury and produces antimicrobial molecules selectively inhibiting skin pathogens. The growth of S. epidermidis is thought to inhibit growth of S. aureus.

Accordingly, the invention provides the use of a topical formulation of the invention for preventing the growth of S. aureus, in particular on the skin of a human or animal.

Again without being bound by theory, it is thought that the topical formulations of the invention prevent the formation of a biofilm of S. aureus or remove such a biofilm once formed, in particular on the skin of a human or animal. Accordingly, the invention provides the use of a topical formulation of the invention for preventing the formation of a biofilm of S. aureus or removing such a biofilm once formed, in particular on the skin of a human or animal.

In another embodiment, the invention provides the use of the topical formulation as a cosmetic. In the cosmetic use of the invention, the topical formulation is not used in therapy. Rather, the formulation is used to improve the appearance of the surface e.g. skin to which the formulation is topically applied.

Another aspect of the present invention is a method for the preparation of the topical formulation comprising the prebiotic oligosaccharide. In one embodiment, the method involves mixing the prebiotic oligosaccharide as defined above, xylitol and the pharmaceutically or cosmetically acceptable carrier, and then isolating the topical formulation.

The topical formulation is for topical application. The topical formulation can be a dermatological formulation.

Preferably the topical formulation is applied to the skin. Accordingly, the topical formulations are typically in liquid or semi-solid form, for example as an emulsion (e.g. an oil-in-water emulsion or a water-in-oil emulsion), an emollient, a paste, a cream, an ointment, a gel, a shampoo, a wipe, a mouthwash, a mouthrinse, or a spray.

Accordingly, the formulation also comprises a pharmaceutically or cosmetically acceptable carrier of the sort suitable for applications intended for topical application.

The topical formulation is preferably an oil-in-water emulsion comprising an oil, at least one surfactant- emulsifier, at least one probiotic oligosaccharide, xylitol, and water.

The oil is generally a mineral oil. The emulsion comprises other ingredients which are typically present in oil-in-water emulsions. For example, the formulation of the invention may optionally contain a moisture protecting agent, such as glycerin. This formulation may also contain a fatty acid alcohol, e.g., stearyl alcohol, palmityl alcohol, maleic alcohol or cetyl alcohol. The present formulation also contains an aqueous medium, e.g. generally purified water.

The fatty acid may have a maximum of 16 to 18 carbons, for example stearate and palmitate, among others. Optional enhancers may be included, for example, citrate, maleic acid, ethylenediaminetetraacetic acid, free amino acids and carbopol, xanthan gum, carrageenan, and conditioning quaternarized polymers. Additionally, it is desirable to include a small amount of preservative in the formulation of the present invention to enable storage of the formulations in good condition. Preservatives such as methyl and propyl paraben may be employed, as well as other known preservatives like, phenoxyethanol, sodium benzoate, and potassium sorbate. Formulations can be preserved using also pentylene glycol, decylene glycol, 1 ,2-octanediol, 1 ,2-hexanediol, and o-Cymen-5-ol.

Additionally, it is desirable to include surfactants in the formulation of the present invention. The formulation can contain sodium laureth sulphate, sodium myreth sulphate, disodium laureth sulfosuccinate, sodium cocoamphoacetate, and sodium cocoamphoacetate. Depending upon the specific formulation of the formulation the other ingredients or excipients will vary in their functionality and amount. For example, in addition to the prebiotic oligosaccharide and xylitol, body wash, body lotion, emollients, body wash and mouthwash formulations typically contain the following amounts of excipients.

Body wash

Shampoo

Body lotion Emollient

Mouthwash

The formulations according to the invention are stable, and exhibit good physical and chemical stability over time, even at a temperature above ambient temperature.

In another embodiment, the topical formulation is supplied on a wipe. In this embodiment, the user can topically apply the formulation using the wipe.

The following examples describe the successful inhibition of S. aureus with the formulations of the invention, and therefore indicate that the topical formulations would be useful for treating skin diseases and disorders, in particular atopic dermatitis. Examples

The reduction in growth of the planktonic and biofilm form of S. aureus and S. epidermidis was investigated using the prebiotic oligosaccharides and xylitol compositions of the invention.

The planktonic form of bacteria is characterized by separated cells that independently float or exist as a suspension within liquid media. The biofilm (or aggregated/sessile) form is characterized by a state in which the cells are tightly constrained and firmly attached to one another. Typically, the biofilm form is associated with clinical inflammation and infection in humans. Materials and methods

Xylitol (XYL), galactooligosaccharides (GOS), fructooligosaccharides (FOS), isomaltooligosaccharides (IMO), arabinogalactan (LAG), inulin and dextran were purchased by Fiberaid Lonza (USA). Each prebiotic compound was prepared in Phosphate-Buffered Saline (PBS, Thermo Fisher scientific, Milan, Italy) and stored at -20°C. The stock dilutions of XYL and GOS were diluted in PBS to obtain the final concentrations of 1 , 2.5, 5% and the stock dilutions of FOS, IMO, LAG, inulin, dextran were diluted in PBS to obtain the final concentrations of 1%.

The Staphylococcus aureus 815 (clinical strain) and Staphylococcus epidermidis 317 (clinical strain) used in this study were sourced from the private collection of the Bacteriology Laboratory of Pharmacy Department. Staphylococcus aureus 815 was previously characterized for some biofilm related properties (see, for example, Microb Biotechnol. 2009; 2(6): 634-641 and J Med. Microbiol., 2007; 56(Pt 4): 519-523) whereas S. epidermidis 317 was analyzed in this study for its hemolytic activity and for the detection of agr allels and icaA/icaD genes. Briefly, for the hemolytic activity, 5 pi of refreshed S. epidermidis 317 broth culture in Tryticase Soy Broth (TSB, Oxoid, Milan, Italy) (OD6oo=0.04-0.05) were spotted onto Trypticase Soy Agar (TSA, Oxoid, Milan, Italy) plus 5% sheep sterile blood (Biomerieux) and incubated for 48 h at 37°C, then chilled at 4°C for 1 h for the evaluation; for the agrgenotyping, the multiplex polymerase chain reaction (PCR) was performed according to Di Stefano et al. (Microb Biotechnol. 2009; 2(6): 634-641) and the presence of icaA, icaD were detected by PCR according to Zhou et al.. (BMC Infect Dis. 2013;13: 242). The characterized strains, stored at -80°C, were cultured in TSB (Oxoid, Milan, Italy) at 37°C overnight under aerobic condition. After incubation, the broth cultures were diluted 1 :10 in the same medium and refreshed for 2 h at 37°C in thermostatic shaking (160 rpm) bath. Finally, the cultures were adjusted in spectrophotometer (Eppendorf, Milan, Italy) to optical density Oϋboo = 0.12 corresponding to 0.5 Mcfarland (Int J Mol Sci. 2013; 14: 13615-13625). These standardized broth cultures were used for the experiments. S. aureus 815 and S. epidermidis 317 were characterized for hemolytic activity, agr alleles, icaA/icaD and biofilm production:

Method 1 - Planktonic growth reduction

The prebiotic oligosaccharides galactooligosaccharides (GOS), fructooligosaccharides (FOS), isomaltooligosaccharides (IMO), arabinogalactan, inulin and dextran were tested in combination with xylitol to assess the antibacterial effect on S. aureus 815 and S. epidermidis 317 in the planktonic phase. 50 mI ofXYL (1%), and 50 mI of the prebiotic oligosaccharide selected from GOS, FOS, IMO, LAG, inulin ordextran (1% respectively) and 100 mI of each standardized broth cultures were inoculated in 96 wells plates and incubated for 3 and 24 h at 37°C in aerobic condition. After each tested time, the Oϋboo was measured with ELISA reader (SAFAS, Munich, Germany). For control, the bacterial strains were incubated without prebiotic formulations. The antibacterial effect of each probiotic oligosaccharide combined with XYL was determined as percentage of Oϋboo reduction in respect to the control. The detection of Oϋboo reduction after 3 and 24 hours was representative of an immediate and effective effect. Each determination was performed in duplicate for three independent experiments.

Method 2 - Biofilm formation reduction The anti-biofilm activity of 1 % XYL combined with 1 % GOS, FOS, IMO, LAG, inulin or dextran was also performed on S. aureus 815 and S. epidermidis 317 biofilms formation. Briefly, 100 mI of each prebiotic combinations (50 mI 1% XYL + 50 mI 1% GOS, FOS, IMO, LAG, inulin or dextran) and 100 mI of standardized broth cultures were inoculated in 96 flat-bottomed microtiter plates and incubated for 3 and 24h at 37°C. After incubation, each well of 96 flat-bottomed microtiter plates, was washed twice with sterile PBS, air-dried, stained for 1 min with 0.1% safranin and washed with PBS. The stained biofilms were resuspended in 200 mI ethanol (95% v/v) and measured at OD492 by spectrophotometry using an ELISA reader (for example, as in Nat Prod Res. 2016; 30: 1870-1874). Each determination was performed in duplicate for three independent experiments. For control, the bacterial strains were incubated without prebiotic formulations. The biofilm formation reduction effect of each probiotic oligosaccharide combined with XYL was determined as percentage of OD ₄₉₂ reduction in respect to the control.

Example 1 : Efficacy of various prebiotic oligosaccharides in combination with xylitol in S. aureus 815 and S. epidermidis 317 planktonic growth reduction in respect to the control

The percentage of planktonic S. aureus 815 and S. epidermidis 317 growth reduction (Oϋboo) after 3, 24 h of contact with 1% of Xylitol (XYL) and 1% of formulations GOS, FOS, IMO, LAG, inulin, dextran was as follows:

‘statistically significant in respect to the control Reference example Example 2: Efficacy of various prebiotic oligosaccharides in combination with xylitol in S. aureus 815 and S. epidermidis 317 biofilm growth reduction in respect to the control

The percentage of S. aureus 815 and S. epidermidis 317 biofilm formation (OD492) reductions after treatment with 1% XYL and 1% GOS, FOS, IMO, LAG, inulin, dextran at 3 and 24 h were as follows:

‘statistically significant in respect to the control Reference example

Example 3: Stability of formulations

Three formulations (Examples 3.1-3.3) were subjected to stability studies.

Each formulation contained a fructooligosaccharide (0.5 %w/w) and xylitol (5.0 %w/w). Depending on formulation type, each formulation contained additional excipients as set out in the description.

The formulations were stored in PET bottles at the temperatures and relative humidity indicated in the tables.

In each case, the aspect, odor, viscosity, pH and pack compatibility (flow rate) of the formulations was analysed.

The aspect was assessed by visual inspection. The odor was assessed by XX. The viscosity was measured using a Brookfield DV2T viscometer (Spindle RV05, 10 rpm). The pH of the formulations was measured over time using a Seven Easy Mettler Toledo pH meter. Pack compatibility (flow rate) was assessed by visual inspection. Example 3.1 Atopic daily cream

Example 3.2 Gentle cleanser Example 3.3 Emoillent