CN118139633A

CN118139633A - Probiotic composition for inhibiting fungal proliferation

Info

Publication number: CN118139633A
Application number: CN202280057910.6A
Authority: CN
Inventors: 夏洛特·韦泽尔; 延斯·爱德华·特斯多普夫; 瑟伦·凯鲁夫; 海伦娜·法尔霍尔特·埃尔维巴克
Original assignee: Leto Biology Co ltd
Current assignee: Leto Biology Co ltd
Priority date: 2021-08-25
Filing date: 2022-08-25
Publication date: 2024-06-04

Abstract

The present invention relates to a composition comprising one or more isolated probiotic strains, two or more active substances in the one or more isolated probiotic strains, or a combination of one or more isolated probiotic strains and two or more active substances in the one or more isolated probiotic strains; and an acceptable carrier, wherein the one or more isolated probiotic strains and/or two or more active substances in the one or more isolated probiotic strains are capable of inhibiting fungal proliferation.

Description

Probiotic composition for inhibiting fungal proliferation

Technical Field

The present invention relates to antimicrobial compositions comprising microbial derived actives and, more particularly, to compositions comprising probiotics or a metazoan fraction produced by probiotics that are effective in inhibiting fungal proliferation such as the proliferation of malassezia furfur (Malassezia furfur) and in treating Seborrheic Dermatitis (SD).

Background

Seborrheic dermatitis is a histopathologically eczematous skin disease characterized by poorly defined scaly erythema with yellowish greasy scales. "dandruff" is a mild form of this condition localized to the scalp. Such diseases may involve any one, several or all of the following: scalp, eyebrows, inter-eyebrows, paranasal and chin folds, ear and posterior auricles, pre-sternal inter-scapular region, pubic region and inter-hip folds.

In some cases, corticosteroids with tars, sulfur or antibiotics can temporarily control the condition, but the condition is often chronic and recurs after termination of treatment. Current therapies include topical and systemic antimicrobial agents, corticosteroids and topical tars. Infectious organisms associated with these skin conditions may spread to other skin areas and may even infect others and may cause changes in the natural skin microflora, causing the microflora to malfunction, further exacerbating the disease. Current therapies include local and systemic antibiotics and antimicrobial drugs. Traditional treatments of seborrheic dermatitis have a significant impact on the scalp, especially the microflora, and therefore require a less severe treatment and the reconstruction of the microflora of the dysfunction associated with seborrheic dermatitis.

Seborrheic dermatitis corresponds to excessive and significant desquamation of the scalp due to excessive rapid proliferation of epidermal cells and their abnormal maturation. A variety of factors may induce such disorders, such as excessive hair treatment, extreme climatic conditions, stress, diet, fatigue or pollution. However, seborrheic dermatitis, and in particular persistent recurrence, is most often caused by a disorder in the scalp microflora, and more specifically, by excessive colonization of fungi belonging to the genus malassezia, the family saccharomyces, which naturally occur on the scalp.

Malassezia (Malassezia) colonizes the skin of a variety of mammals, domestic animals and birds. These lipophilic yeasts inhabit the stratum corneum of human and animal skin because this layer is rich in lipids. For reasons not known at present, these yeasts can change their saprophytic state and invade the stratum corneum as a pathogen.

Infant scab is a form of seborrheic dermatitis. Infant scalp with scab has slight red scale or scab-like yellow patches. It may also start from the face or diaper area and spread to other parts of the body.

Malassezia (Malassezia) also infects the skin of animals and is common in dermatitis in horses, cattle, sheep, dogs, and cats. Malassezia (Malassezia) is usually present on the skin, but its abnormal overgrowth can cause dermatitis, or inflammation of the skin, ears, mouth and body ducts.

Seborrheic dermatitis, including milder symptoms commonly known as dandruff, affects up to 50 percent of the world's population, and affects both men and women, often with negative psychological effects. Dandruff is aesthetically unpleasant (due to the apparent presence of dead skin) and because individuals feel personal discomfort (particularly itching). Thus, an affected person facing this problem to varying degrees would like to get it out effectively and permanently.

Despite the many commercially available therapies, scalp treatments and shampoos, seborrheic dermatitis remains a challenging condition, and many patients do not respond to several attempted therapies, making the treatment unpredictable and difficult to take in many cases.

The main objective of many anti-dandruff treatments is to eradicate malassezia yeasts from the scalp. Thus, the activity of today's anti-dandruff actives such as zinc pyrithione, piroctone olamine, climbazole (climbazole), ketoconazole (ketoconazole) or selenium sulphide is mainly based on their fungicidal properties. More recently, other formulations have been described using natural based alternatives, such as an anti-dandruff composition based on ellagic acid and at least one essential oil as described in WO2011/138450, or a hydrolysable tannin-rich active derived from punica granatum as described in FR 2908045.

Despite the above, there remains a need for new effective natural solutions to inhibit fungal proliferation and/or to treat or prevent mycoses and/or to treat or prevent dandruff that do not cause skin irritation and that will promote the reestablishment of a balanced natural scalp microflora.

Disclosure of Invention

The present invention is premised on the recognition that various combinations of anti-dandruff actives derived from probiotic microorganisms are formulated in, inter alia, topical compositions that still include a microbial derived active against Seborrheic Dermatitis (SD), and more particularly to compositions comprising probiotics or a metazoan fraction produced by probiotic microorganisms that are effective in inhibiting fungi having the genus malasseziaceae, such as malassezia furfur (Malassezia furfur), and in treating seborrheic dermatitis.

One aspect of the invention relates to a composition comprising:

-one or more isolated probiotic strains; two or more active substances in the one or more isolated probiotic strains; or a combination of one or more isolated probiotic strains and two or more active substances in said one or more isolated probiotic strains, and

An acceptable carrier, which is a mixture of the components,

Wherein the one or more isolated probiotic strains and/or two or more active substances in the one or more isolated probiotic strains are capable of inhibiting fungal proliferation.

Another aspect of the invention relates to an anti-dandruff composition comprising (or consisting essentially of) the following: a combination of at least two anti-dandruff actives derived from probiotic microorganisms. The anti-dandruff active is present in the anti-dandruff composition in an effective amount to treat or prevent dandruff.

Yet another aspect of the invention relates to the use in the prevention and/or treatment of mycoses in humans or animals.

The present invention also seeks to provide compounds in hair care formulations which may provide comparable or improved antifungal properties, such as anti-dandruff properties, as compared to existing anti-dandruff agents.

One aspect of the present invention relates to a topical hair care composition comprising a composition according to the present invention in combination with a shampoo base having a pH of less than 6.5, said shampoo base comprising at least one detersive surfactant selected from the group consisting of: anionic surfactant, nonionic surfactant, amphoteric surfactant, or combinations thereof.

One aspect of the invention relates to leave-on compositions for the treatment of mycoses.

The invention also seeks to provide the use of a compound as an anti-dandruff agent, and a formulation comprising said compound for reducing dandruff on human skin.

According to another embodiment of the present invention, there is provided an anti-dandruff hair care composition suitable for the treatment of dandruff against malassezia furfur (Malassezia furfur).

The composition according to the invention may comprise one or more isolated probiotic strains and/or two or more actives in one or more isolated probiotic strains, such as an anti-dandruff active, an acceptable carrier, and have a pH of less than 6.5.

Surprisingly, it was determined that the synergistic effect between the antifungal effect (e.g. anti-dandruff active) and the pH of the composition. Without being bound by theory, it is believed that this synergy is identified as at least two different mechanisms, and is observed for live probiotic microorganisms and any fraction thereof that still includes antifungal effects, particularly anti-dandruff actives. Thus, a synergistic effect between the antimicrobial activity of the active substance and the inhibition of fungi may have been observed, which may be a fungus of the genus malasseziaceae, such as malassezia furfur (Malassezia furfur), and a synergistic effect between the pH of the anti-dandruff composition and the ability of the natural microflora to re-establish after treatment, and thereby allowing a gentle anti-fungal treatment, such as a gentle anti-dandruff treatment, which does not repair or promote repair of the microflora of the dysfunction left on the scalp after the antimicrobial treatment, which has a longer, more durable effect than conventional treatments.

As a further advantage of the present invention, the probiotic treatment does not cause any irritation to the human or animal body or skin or scalp.

According to yet another embodiment of the present invention, there is provided a method for treating a dandruff condition associated with proliferation of malassezia yeasts on the scalp of a subject. The method comprises applying an effective amount of an anti-dandruff composition to the scalp of a subject, wherein the effective amount of the anti-dandruff composition inhibits proliferation of malassezia yeasts on the scalp.

The present invention will be described in more detail below.

Detailed Description

The present invention may relate to compositions, uses of the compositions, and methods for inhibiting fungal proliferation, preventing or treating scalp disorders associated with dysfunctional microflora. The composition, e.g. an anti-dandruff composition, may comprise functional probiotics and/or metabolites obtained by fermentation of the probiotics.

Anti-dandruff compositions, particularly shampoos, are well known and have been commercially available for many years. Many anti-dandruff actives have been used commercially, such as ketoconazole, zinc pyrithione, piroctone olamine, pyridone olamine, salicylic acid, selenium sulfide, coal tar, and azelaic acid.

These types of actives may preferably be active compounds that generally act as antimicrobial/fungicidal agents that are effective against certain species and strains of fungi, yeasts and/or bacteria, in particular against fungi and/or yeasts. For example, the yeast like fungus malassezia lives on the scalp of most adults, but for some it stimulates the scalp and can cause more skin cell growth. Although malassezia are part of the normal microflora, they can cause superficial skin infections under certain conditions. These unwanted skin cells die and shed, making them appear white and flaky in hair and on clothing. Thus, substances active against malassezia, in particular the species malassezia furfur, can reduce the severity of dandruff.

Preferably, one or more active substances in one or more probiotic strains (preferably isolated probiotic strains) may be provided in an amount effective to inhibit fungal proliferation.

These shampoos and other subject antifungal formulations are often combined with corticosteroids to control inflammation and reduce pain and itching. However, the use of these molecules may not produce satisfactory results, and in some cases, these compounds exhibit inherent and undesirable cytotoxicity and damage to the microflora.

There is a continuing need for improved anti-dandruff actives and end use products containing anti-dandruff actives that are non-irritating to the skin and have no damaging effect on the scalp microflora.

When administered topically, the present invention may provide effective inhibition, treatment or prevention without damaging effects on the natural microflora.

A preferred embodiment of the present invention relates to a composition comprising:

An acceptable carrier, which is a mixture of the components,

Preferably, the fungus capable of fungal proliferation may be a fungus selected from the genus malasseziaceae. Preferably, the genus from the malasseziaceae family may be malassezia.

Another preferred embodiment of the invention relates to a composition (e.g. an anti-dandruff composition) comprising an effective amount of a probiotic microorganism or a fraction of a probiotic microorganism capable of producing an active substance (e.g. an anti-dandruff active), wherein the fraction comprises the active substance.

According to another aspect of the present invention there is provided a method of providing antifungal efficacy, in particular anti-dandruff efficacy, comprising the steps of:

(i) Wetting hair with water;

(ii) Administering an effective amount of a composition of the present invention comprising a probiotic microorganism or fraction thereof

(Iii) Rinsing the composition on the hair with water; and

(Iv) Optionally repeating steps (ii) and (iii).

According to another aspect of the present invention there is provided a method of providing antifungal efficacy, the method comprising the steps of: leave-on compositions are applied to the skin, epithelium, nails, oral cavity, including the scalp, feet, vagina, genitals, and the skin of the ear.

According to a further aspect of the present invention there is provided a method for killing or delaying the growth of a fungus, in particular a fungus from the family malasseziaceae, such as Malassezia spp, the method comprising the step of contacting the fungus, such as Malassezia, with a composition according to the invention comprising an active substance derived from a probiotic microorganism.

As used herein, the terms "e.g." (for example) "," e.g. "(for instance)", "such as" or "comprising" are intended to introduce examples that further clarify more general subject matter. These examples are provided solely as an aid in understanding the application exemplified in the present disclosure, and are not meant to be limiting in any way, unless otherwise specified.

The term "antifungal composition" relates to a composition according to the invention capable of inhibiting fungal proliferation, treating or preventing fungal infections, for example treating or preventing mycoses and/or dandruff in humans or animals.

Even though the anti-dandruff effect is the effect mainly described in the present invention, the composition according to the present invention shows a strong antifungal effect.

Fungal proliferation may lead to dandruff and/or mycosis in mammals. By limiting, reducing or stopping fungal proliferation, it is believed that the occurrence of dandruff and/or mycoses may be prevented, avoided or limited.

Preferably, the composition according to the invention may be an anti-dandruff composition and/or an anti-fungal composition.

The term "anti-dandruff composition" refers to an agent that provides a prophylactic and/or therapeutic effect on scalp dandruff. This includes preventing and/or reducing excessive dandruff formation, and/or visually unattractive excessive dandruff formation.

The term "active substance" may relate to a compound having antifungal proliferative activity for the treatment and/or prevention of mycoses in humans or animals. The active substance may preferably comprise bacteriocins, organic acids, cell wall substances or combinations thereof.

The present invention relates to antifungal compositions, particularly anti-dandruff compositions, and methods of using the compositions to treat or inhibit antifungal proliferation, such as for treating or inhibiting dandruff. The composition according to the invention is useful for application to keratinous tissue or scalp surfaces and comprises an effective amount of a combination of active agents that inhibits, reduces or eliminates symptoms caused by the proliferation of malassezia furfur, such as dandruff symptoms. The compositions of the present invention may be in a variety of product forms including, but not limited to, solutions, suspensions, lotions, creams, gels, ointments, oils, emulsions, sprays, aerosols, shampoos, hair conditioners, pastes, foams, powders, mousses, tissues, strips, patches, hydrogels, film-forming products, and the like. The composition form may be based on the particular dermatologically acceptable carrier selected.

In embodiments of the present invention, the anti-dandruff composition may comprise a composition applied to hair and/or skin underlying hair, and the anti-dandruff composition may comprise (or consist essentially of) at least one of the following various combinations: the combination of an active substance derived from a probiotic and a dermatologically acceptable carrier is effective in inhibiting malassezia furfur and in treating dandruff.

An active agent may refer to a single compound or a composition comprising two or more compounds which, when present in an effective amount in the composition, has the ability to inhibit the proliferation of fungi such as malassezia furfur.

The term "effective amount" as used herein means an amount of a compound or a composition comprising two or more compounds sufficient to reduce or inhibit the proliferation of fungi, particularly malassezia furfur, reduce or inhibit the visible and/or physical effects of fungal infection, mycosis, or dandruff caused by fungal proliferation (such as proliferation of malassezia furfur), or reduce or inhibit scalp itching in a statistically significant amount.

The term "topical application" as used herein means the application or spreading of the composition of the present invention to the surface of hair, skin, such as the scalp.

The term "acceptable carrier" as used herein means a carrier suitable for internal (e.g., oral or rectal ingestion) or external (topical) use without undue toxicity, incompatibility, instability, allergic response, discomfort, and the like.

The term "dermatologically acceptable" as used herein means that the compositions so described, or components thereof, are suitable for use in contact with mammalian keratinous or skin tissue without undue toxicity, incompatibility, instability, allergic response, and the like.

According to an embodiment of the invention, the composition, e.g. the anti-dandruff composition, comprises (or consists essentially of) the following: an effective amount of a combination of at least two active substances selected from the group of organic acids. In a further preferred embodiment, the organic acid is selected from the group consisting of: lactic acid, citric acid, acetic acid, malic acid, tartaric acid, 3-phenyllactic acid, 3-hydroxyphenyllactic acid, 4-hydroxyphenyllactic acid, propionic acid and succinic acid, salicylic acid, azelaic acid, indole-3-lactic acid, indole-3-acetic acid, 2-hydroxybutyric acid, 2-hydroxyisohexanoic acid, 3- (R) -hydroxydecanoic acid, 3-hydroxy-5-cic-dodecanoic acid, 3- (R) -hydroxydodecanoic acid, 3- (R) -hydroxytetradecanoic acid, glycolic acid and N-acetoacetic acid.

In embodiments of the invention, the composition may comprise viable probiotic microorganisms. In another embodiment of the invention, the active substance present in the composition may be in the form of a live probiotic microorganism. Preferably, the live probiotic microorganism may be capable of inhibiting fungal proliferation, such as inhibiting malassezia furfur. Preferably, inhibition of malassezia furfur is obtained when the composition can be topically applied to the scalp or skin area affected by dandruff.

The term "viable" or "viable" as used herein relates to a microorganism that is not dead and is capable of having active metabolism.

The term "microflora" as used herein relates to communities of commensal, symbiotic and pathogenic microorganisms found in and on all multicellular organisms. The microflora include bacteria, archaea, protists, fungi, yeasts, viruses and phages.

The term "dysfunctions of a microflora" as used herein relates to a state in which the microflora is not functioning properly or functioning is completely hindered. Unless otherwise indicated, the present invention contemplates that a dysfunction of a microflora includes overgrowth or increased growth of pathogenic microorganisms of the microflora that caused the dysfunction. An example of a microflora dysfunction is an increase in malassezia furfur, leading to mycoses or seborrheic dermatitis, such as dandruff.

In embodiments of the invention, the fungal infection may be a mycosis or seborrheic dermatitis. Preferably, the seborrheic dermatitis may be dandruff.

The term "probiotic microorganism" as used herein relates to a living microorganism which is intended to have a health benefit when consumed by or administered to a host. Examples of suitable probiotic microorganisms include yeasts such as Saccharomyces (Saccharomyces), debaryomyces (Debaryomyces), candida (Candida), pichia (Pichia) and Torulopsis (Torulopsis), mold such as Aspergillus (Aspergillus), rhizopus (Rhizopus), mucor (Mucor) and Penicillium (Penicillium) and Torulopsis (Torulopsis), bacteria such as Bifidobacterium (Bifidobacterium), bacteroides (Bacteroides), clostridium (Clostridium), clostridium (Fusobacterium), apis (Melissococcus), propionibacterium (Propionibacterium), streptococcus (Streptomyces), enterococcus (Enterococcus), lactobacillus (Lactobacillus), staphylococcus (Staphylococcus), streptococcus (Streptococcus), streptococcus (Peptostrepococcus), bacillus (Bacillus), pediococcus (Pediococcus), micrococcus (Micrococcus), leuconostoc (Leuconostoc), scolopendra (Wesela), bacillus (Aerococcus), lactobacillus (Lactobacillus) and Lactobacillus (Lactobacillus plantarum) (5257), lactobacillus (Lactobacillus plantarum) and Lactobacillus (Lactobacillus plantarum (5257).

The most commonly used probiotics are strains of Lactic Acid Bacteria (LAB). The term "lactic acid bacteria" includes species from the following families: lactobacillaceae (Lactobacillaceae), balloon bacteriaceae (Aerococcaceae), bifidobacteriaceae (Bifidobacteriaceae), sarcobacteriaceae (Carnobacteriaceae), enterococcaceae (Enterococcaceae), leuconostoc (Leuconostocaceae) and Streptococcaceae (Streptococcaceae). These are considered to be nonpathogenic and are generally used as probiotics to improve the gastrointestinal flora and treat gastrointestinal symptoms. The present invention relates to topical administration of probiotics characterized in that the probiotics produce anti-dandruff actives.

The microorganism may preferably be a lactic acid bacterium. The microorganism may even more preferably be selected from the following genera: lactobacillus (Lactobacillus), lactobacillus plantarum (Lactiplantibacillus), lactobacillus helveticus (Holzapfelia), lactobacillus amylovorus (Amylolactobacillus), lactobacillus bufonis (Bombilactobacillus), lactobacillus companion (Companilactobacillus), lactobacillus la Pi Di (Lapidilactobacillus), lactobacillus agricultural (Agrilactobacillus), lactobacillus Shi Laifu (Schleiferilactobacillus), lactobacillus loyi (Schleiferilactobacillus), lactobacillus paracasei (Schleiferilactobacillus), lactobacillus sake (Schleiferilactobacillus), schleiferilactobacillus, lactobacillus glycyrrhiza (Schleiferilactobacillus), lactobacillus hexa-complex (Schleiferilactobacillus), lactobacillus furfur (Schleiferilactobacillus), lactobacillus oligosacchari (Schleiferilactobacillus), lactobacillus mucilaginosa (Schleiferilactobacillus), lactobacillus mucrobial (Schleiferilactobacillus), lactobacillus fruit (Schleiferilactobacillus), lactobacillus plantarum (Schleiferilactobacillus) Lactobacillus (Schleiferilactobacillus), lactobacillus melissa (Schleiferilactobacillus), lactobacillus progenitalis (Schleiferilactobacillus), lactobacillus paracasei (Schleiferilactobacillus), lactobacillus lentus (Schleiferilactobacillus), leuconostoc (Leuconostoc), bifidobacterium (bifidobacteria), pediococcus (Pediococcus), lactococcus (Lactococcus), streptococcus (Streptococcus), balloon (Aerococcus), clostridium (Schleiferilactobacillus), enterococcus (Enterococcus), enterococcus (Schleiferilactobacillus), lactobacillus (Schleiferilactobacillus), tetracoccus (Schleiferilactobacillus), roaming bacteria (Schleiferilactobacillus) and Weissella (Weissella).

Preferred microorganisms may in particular be bacteria. The probiotics may preferably be selected from the group comprising: lactococcus lactis (Lactococcus lactis), lactobacillus rhamnosus (Lacticaseibacillus rhamnosus), lactobacillus plantarum (Lactiplantibacillus plantarum), lactobacillus helveticus (Lactobacillus helveticus), lactobacillus jensenii (Lactobacillus jensenii), lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus bulgaricus (Lactobacillus bulgaricus), lactobacillus amylovorus (Lactobacillus amylovorus), lactobacillus amyloliquefaciens (Lactobacillus amylolyticus), lactobacillus plantarum (Lactobacillus alimentarius), lactobacillus avium (Lactobacillus aviaries), lactobacillus delbrueckii (Lactobacillus delbrueckii), lactobacillus saccharolyticus (Lactobacillus diolivorans), Lactobacillus farnesii (Lactobacillus farciminis), lactobacillus gallinarum (Lactobacillus gallinarum), lactobacillus casei (Lacticaseibacillus casei), lactobacillus crispatus (Lactobacillus crispatus), lactobacillus gasseri (Lactobacillus gasseri), lactobacillus johnsonii (Lactobacillus johnsonii), lactobacillus johnsonii (Lactobacillus hilgardii), Lactobacillus kefir matrix (Lactobacillus kefiranofaciens), lactobacillus kefir (Lactobacillus kefiri), lactobacillus mucosae (Lactobacillus mucosae), lactobacillus pani (Lactobacillus panis), lactobacillus paraphytosis (Lactiplantibacillus paraplantarum), lactobacillus Pang Diru (Lactobacillus Pontis), lactobacillus sake (Latilactobacillus sakei), Lactobacillus salivarius (Lactobacillus saliverius), lactobacillus sanfranciscensis (Lactobacillus sanfraciscensis), lactobacillus paracasei (Lacticaseibacillus paracasei), lactobacillus pentosus (Lactobacillus pentosus), lactobacillus cellobiose (Lactobacillus cellobiosus), lactobacillus Ke Linshi (Lactobacillus collinoides), Coryneform bacterium (Lactobacillus coryniformis), lactobacillus curvatus (Lactobacillus curvatus), lactobacillus brevis (Levilactobacillus brevis), lactobacillus buchneri (Lactobacillus buchneri), lactobacillus fructoses (Lactobacillus fructivorans), lactobacillus hilgardii (Lactobacillus hilgardii), lactobacillus fermentum (Lactobacillus fermentum), Lactobacillus reuteri (Lactobacillus reuteri), lactobacillus anguillarum (Lactobacillus ingluviei), weissella viridis (WEISSELLA VIRIDESCENS), bifidobacterium bifidum (Bifidobacterium bifidum), bifidobacterium adolescentis (Bifidobacterium adolescentis), bifidobacterium breve (Bifidobacterium breve), bifidobacterium longum (Bifidobacterium longum), Bifidobacterium animalis (Bifidobacterium animalis), clostridium cloacae (Carnobacterium divergens), corynebacterium glutamicum (Corynebacterium glutamicum), leuconostoc citreum (Leuconostoc citreum), leuconostoc lactis (Leuconostoc lactis), leuconostoc mesenteroides (Leuconostoc mesenteroides), leuconostoc pseudomesenteroides (Leuconostoc pseudomesenteroides), The composition comprises the components of wine coccus (Oenococcus oeni), pasteurella (Pasteuria nishizawae), pediococcus acidilactici (Pediococcus acidilactici), pediococcus dextrin (Pediococcus dextrinicus), pediococcus parvulus (Pediococcus parvulus), pediococcus pentosaceus (Pediococcus pentosaceus), propionibacterium freudenreichii (Probionibacterium freudenreichii), Acid propionic acid probiotics (Probionibacterium acidipropoinici), enterococcus faecium (Enterococcus faecium), enterococcus faecium (Enterococcus faecalis), streptococcus thermophilus (Streptococcus thermophilus), bacillus amyloliquefaciens (Bacillus amyloliquefaciens), bacillus atrophaeus (Bacillus atrophaeus), bacillus clausii (clausii), Bacillus coagulans (Bacillus coagulans), bacillus curvatus (Bacillus flexus), bacillus fusiformis (Bacillus fusiformis), bacillus lentus (Bacillus lentus), bacillus licheniformis (Bacillus licheniformis), bacillus megaterium (Bacillus mega-terium), bacillus mojavensis (Bacillus mojavensis), bacillus pumilus (Bacillus pumilus), Bacillus smithii (Bacillus smithii), bacillus subtilis (Bacillus subtilis), bacillus cereus (Bacillus vallismortis), geobacillus stearothermophilus (Geobacillus stearother-mophilus) or mutants thereof.

In a preferred embodiment of the invention, the composition comprises at least one strain selected from the group consisting of: lactobacillus plantarum LB356R (DSM 33094), lactobacillus plantarum LB244R (DSM 32996), wegener viridis LB10G (DSM 32906), lactobacillus paracasei LB113R (DSM 32907), lactobacillus paracasei LB116R (DSM 32908), lactobacillus brevis LB152G (DSM 32995), lactobacillus paracasei LB28R (DSM 32994), enterococcus faecium LB276R (DSM 32997), leuconostoc mesenteroides (Leuconostoc mesenteriodes) LB349R (DSM 33093), lactobacillus plantarum LB316R (DSM 33091), lactobacillus plantarum LB312R (DSM 33098), pediococcus pentosaceus LB606R (DSM 33730), lactobacillus plantarum LB679R (DSM 33731), lactobacillus crispatus LB714R (DSM 33732), lactobacillus griseus LB905R (DSM 34094), lactobacillus crispatus LB912R (DSM 34095), lactobacillus jensenii LB918R (DSM 34096), lactobacillus crispatus LB919R (DSM 34097), subspecies (Lacticaseibacillus paracasei) and lactobacillus subspecies (DSM 34249) R555; and/or any combination thereof or mutant strains thereof and/or cell lysates and/or soluble metabolites of any of these probiotic strains.

The number of microorganisms is measured in Colony Forming Units (CFU) per ml or gram.

The microorganism according to the invention may preferably be in isolated or purified form, wherein the term "isolated" particularly means that the lactic acid bacteria originate from its medium, e.g. including its natural medium. The term "isolated" or "purified" may not be limited to absolute purity. The terms "isolated" and "purified" are used interchangeably.

In an embodiment of the invention, the probiotic bacterial strain may be used in a stable form as a viable isolated microorganism. Suitable stabilization methods are known to those skilled in the art and include lyophilization or freeze-drying involving different cryoprotectants.

In another embodiment of the invention, the strain may be used as a live isolated strain.

Preferably, the strain can be used as a live isolated strain. Preferably, the strain can be used as a viable isolated stable strain. Even more preferably, the strain may be used as a live isolated strain stabilized by lyophilization. Even more preferably, the strain may be used as a live isolated strain that is stabilized by lyophilization and includes a cryoprotectant.

In a preferred embodiment of the invention, the probiotic bacterial strain is a gram positive bacterium.

In a preferred embodiment of the invention, the composition comprises at least one strain selected from the group consisting of: lactobacillus plantarum LB356R (DSM 33094), lactobacillus plantarum LB244R (DSM 32996), wegener viridis LB10G (DSM 32906), lactobacillus paracasei LB113R (DSM 32907), lactobacillus paracasei LB116R (DSM 32908), lactobacillus brevis LB152G (DSM 32995), lactobacillus paracasei LB28R (DSM 32994), enterococcus faecium LB276R (DSM 32997), leuconostoc mesenteroides LB349R (DSM 33093), lactobacillus plantarum LB316R (DSM 33091), lactobacillus plantarum LB312R (DSM 33098), pediococcus pentosaceus LB606R (DSM 33730), lactobacillus plantarum LB679R (DSM 33731), lactobacillus crispatus LB714R (DSM 33732), lactobacillus griseus LB905R (DSM 34094), lactobacillus crispatus LB 152R (DSM 34095), lactobacillus jensenii LB918R (DSM 34096), lactobacillus helveticus LB919R (DSM 34097), lactobacillus paracasei subspecies (DSM 34249) R (DSM 34249), lactobacillus pentosus R (DSM 34250) or a mutant strain thereof.

The term "metazoan" refers to a compound, metabolite, or cellular material secreted or released by a probiotic microorganism that provides a health benefit when administered to a host. The metacomposition is characterized by having a health benefit in the absence of viable microorganisms.

The term "prebiotic fraction of a probiotic microorganism" as used herein discloses a fermented composition of probiotic microorganisms that is substantially free of living microorganisms. The composition may include a cellular material comprising dead cells.

In embodiments of the invention, the composition may have the following pH: below pH 8.0, such as below pH 7.5, for example below pH 7.0, such as below pH 6.5, for example below pH 6.0, such as below pH 5.5, for example below pH 5.0, such as below pH 4.75, for example below pH 4.5, such as below pH 4.25, for example below pH 4.0, such as below pH 3.75, for example below pH 3.5, such as below pH 3.25.

The preferred pH of the composition is from pH 2.5 to pH 7, more preferably from pH 3 to pH 6.5, and even more preferably from pH 3.5 to pH 5.5. The low pH of the composition resulting from the acid produced by the probiotic microorganisms will cause acidification of the skin surface with an elevated pH when applied to the scalp. Healthy scalp has a pH of about 4.5 and metabolites produced by probiotic microorganisms will act as another beneficial effect, helping to maintain a healthy pH of the skin.

In an embodiment of the invention, at least two active substances may be produced by metabolism of an isolated live probiotic strain.

In another embodiment of the invention, at least two active substances may be produced by a single isolated probiotic strain.

Preferably, the active substance may be selected from bacteriocins, organic acids, cell wall substances or combinations thereof.

In a preferred embodiment of the invention, the active ingredient as an organic acid is present in protonated form as an acid and the pH is equal to or lower than the pKa of the acid.

The present invention is based on the discovery that certain lactic acid bacteria species will produce bacteriocins in the supernatant in amounts effective to inhibit the growth of malassezia, even if the lactic acid bacteria are no longer present.

In one embodiment of the invention, the preferred microorganism is an isolated wild-type lactic acid bacterium.

In one embodiment, the preferred bacteriocins used in the present invention are produced by probiotics, in another preferred embodiment bacteriocins are produced by lactic acid bacteria, in another preferred embodiment bacteriocins are produced by one of the following bacteria; lactobacillus plantarum LB356R (DSM 33094), lactobacillus plantarum LB244R (DSM 32996), wegener viridis LB10G (DSM 32906), lactobacillus paracasei LB113R (DSM 32907), lactobacillus paracasei LB116R (DSM 32908), lactobacillus brevis LB152G (DSM 32995), lactobacillus paracasei LB28R (DSM 32994), enterococcus faecium LB276R (DSM 32997), leuconostoc mesenteroides LB349R (DSM 33093), lactobacillus plantarum LB316R (DSM 33091), lactobacillus plantarum LB312R (DSM 33098), pediococcus pentosaceus LB606R (DSM 33730), lactobacillus plantarum LB679R (DSM 33731), lactobacillus crispatus LB714R (DSM 33732), lactobacillus griseus LB905R (DSM 34094), lactobacillus crispatus LB 152R (DSM 34095), lactobacillus jensenii LB918R (DSM 34096), lactobacillus helveticus LB919R (DSM 34097), lactobacillus paracasei subspecies (DSM 34249) R (DSM 34249), lactobacillus pentosus 34250 and any combination thereof.

Bacteriocins are preferably used in the compositions of the present invention in amounts between 1 and 1,000,000 Arbitrary Units (AU) of bacteriocins. AU of bacteriocin was defined as the 5 microliter highest dilution of culture supernatant that produced a well-defined growth-inhibiting zone with a lawn of the indicator strain on an agar plate.

In embodiments of the invention, the active substance may be a bacteriocin.

The term "bacteriocin" refers to an antimicrobial peptide or protein produced by a bacterium that is active against the microorganism but does not harm the producing bacterium. For the purposes of the present invention, bacteriocins or sources of bacteriocins generally include antimicrobial agents suitable for use as cosmetic or pharmaceutical preparations. Particularly preferred antimicrobial agents include "lanthioids" (i.e., polypeptides containing lanthionine and β -methyl lanthionine). Non-limiting examples of such lanthionines are nisin, such as nisin a or nisin Z, or nisin analogs or related lanthionine-containing peptides, such as pediocin, lactin (lactosin), nisin (e.g., nisin a, nisin B, nisin F), card Mo Su (camocin), enterobacterin, plantaricin, subtilin, epidermin, cinnamycin, duramycin, angiotensin converting enzyme inhibitory peptide (ancovenin), pep 5, and the like, alone or in any combination thereof. Other bacteriocins useful in the present invention include, for example, lactococcus (e.g., lactococcus A, lactococcus B, lactococcus M), leucovorin, helvetican, acidophilin, casein, salivary strept (salivarcin) X, nisin J46, nisin 481, nisin 3147, salivary strept A2, salivary strept A3, salivary strept A4, BHT-Aa, BHT Ab, salivary strept A5, salivary strept B, nisin Al, nisin, Streptococcal A-FF22, streptococcal BNY, streptococcal 1140, streptococcal K8, streptococcal II, smbAB, lanthocin (bovicin) HJ50, lanthocin HC5, maculon (macedocin), leucovorin C, lactobacillus sake 5X, enterobacterin CRL 35/Meng Dimei, avermectin (avicin) A, meng Dimei I, enterobacterin HF, bavarian (bavaricin) A, tuberculin (ubericin) A, leucovorin Y105, leucovorin A, and pharmaceutical compositions containing them, Rice wine lactobacillin G, zedoary turmeric extract A/Rice wine lactobacillin A, lactobacillin (lactocin) 5, lysin (cyctolysin), enterobacterin A, tetrandrine (divercin) V41, tetrandrine M35, bavarian, coagulin, pediocin PA-1, meng Dimei, stigmain (piscicocin) CS526, stigmain 126/Vla, rice wine lactobacillin, pneumobacillin (Pcarnobacteriocin) BM1, enterobacterin P, stigmain Vlb, pan Nuosu (penocin) A, Bacteriocin 31, bacteriocin RC714, sea cocoa (hiracin) JM79, bacteriocin T8, enteromorph SE-K4, botulinum toxin (carnobacteriocin) B2, and plantaricin.

The term "phytochemicals" refers to bacteriocins from lactobacillus plantarum (Lactiplantibacillus plantarum), the main types of phytochemicals including: lactobacillus plantarum A, lactobacillus plantarum E, lactobacillus plantarum F, lactobacillus plantarum J, lactobacillus plantarum K, lactobacillus plantarum C, lactobacillus plantarum D, lactobacillus plantarum W, lactobacillus plantarum T and Lactobacillus plantarum S. Other plant lactobacillins, for example, plant lactobacillin 35d, plant lactobacillin MG, plant lactobacillin 423, plant lactobacillin 154, plant lactobacillin 149, plant lactobacillin 163, plant lactobacillin LC74, plant lactobacillin K25, plant lactobacillin ST31, plant lactobacillin SA6. In particular broad spectrum plantaricins such as plantaricin F, plantaricin DL3, plantaricin ZJ008, plantaricin MG, plantaricin Q7, plantaricin KL-1Y, plantaricin 163, plantaricin 154.

Thus, embodiments of the present invention relate to compositions comprising probiotic microorganisms producing at least one active substance, wherein the active substance comprises (consists essentially of) bacteriocins.

In yet another embodiment of the present invention, at least 2 different bacteriocins may be produced by a probiotic microorganism.

The compositions of the invention comprise probiotic microorganisms capable of producing an active substance, such as an anti-dandruff active, or a fraction from the metazoan of a probiotic organism, wherein the active substance is retained in the metazoan composition.

In embodiments of the invention, the active substance may comprise an organic acid, or a combination of one or more organic acids and one or more bacteriocins.

The organic acid is preferably used in the metacomposition at a total concentration of about 0.1 to 20% by weight. For example, 1 to 5% by weight of lactic acid and acetic acid. The organic acid is preferably selected from lactic acid, citric acid, acetic acid, malic acid, tartaric acid, phenyllactic acid, 3-hydroxyphenyllactic acid, 4-hydroxyphenyllactic acid, propionic acid and succinic acid, salicylic acid, azelaic acid, indole-3-lactic acid, indole-3-acetic acid, 2-hydroxybutyric acid, 2-hydroxyisohexanoic acid, 3- (R) -hydroxydecanoic acid, 3-hydroxy-5-cic-dodecanoic acid, 3- (R) -hydroxydodecanoic acid, 3- (R) -hydroxytetradecanoic acid, glycolic acid and N-acetoacetic acid, preferably used in a concentration of 0.1 to 10% in the metacomposition.

As described above, the active portion of the composition comprises (or consists essentially of) a combination of at least two active substances, which are derived from or administered in the form of a viable probiotic microorganism capable of producing at least two active substances.

In embodiments of the invention, the active substance may be produced by a probiotic, and the active substance may be selected from; lactic acid, citric acid, acetic acid, malic acid, tartaric acid, 3-phenyllactic acid, 3-hydroxyphenyllactic acid, 4-hydroxyphenyllactic acid, propionic acid and succinic acid, salicylic acid, azelaic acid, indole-3-lactic acid, indole-3-acetic acid, 2-hydroxybutyric acid, 2-hydroxyisohexanoic acid, 3- (R) -hydroxydecanoic acid, 3-hydroxy-5-cic-dodecanoic acid, 3- (R) -hydroxydodecanoic acid, 3- (R) -hydroxytetradecanoic acid, glycolic acid and N-acetoacetic acid.

The active may be produced on the scalp by probiotics and may be selected from; 2-hydroxyisocaproic acid, phenyllactic acid, salicylic acid, acetylsalicylic acid, indole-3-lactic acid, gallic acid, azelaic acid, 2-hydroxybutyric acid, N-acetoacetaspartic acid, succinic acid and lactic acid.

In embodiments of the present invention, the composition may include at least one of the following active substance combinations: bacteriocins and/or phenyllactic acid and/or salicylic acid and/or lactic acid and/or 2-hydroxyisocaproic acid and/or azelaic acid and/or succinic acid and/or indole-3-lactic acid and/or 2-hydroxybutyric acid and/or N-acetoacetic acid and/or indole-3-acetic acid and/or gallic acid.

In another embodiment of the invention, the active substance may comprise a combination of at least 2 different bacteriocins produced by or isolated from a fraction fermented by a probiotic microorganism, wherein the probiotic microorganism has not been genetically modified to produce bacteriocins.

The present invention may be suitable for use directly as a scalp care product or formulated as a scalp care product in a therapeutic or scalp care composition for preventing or treating scalp disorders or for modulating microflora dysfunction.

The probiotic microorganisms of the present invention may be capable of producing a yield of functional metabolites, e.g. actives, such as anti-dandruff actives, sufficient to provide a broad spectrum of antimicrobial activity, anti-inflammatory activity, peeling, moisturizing and/or functional effects, such as on foot, nail and scalp hardness, during growth. Without being bound by theory, it is believed that this effect may be provided by activation of fibrillar protein and collagen synthesis.

Furthermore, it is surprising to determine the synergy between the active or metabolite present in the composition and the pH of the composition. This synergy may allow for functional concentrations of each active to be lower than those required for the purified active.

For example azelaic acid may be used in high concentrations. In embodiments of the invention, azelaic acid may be used at a concentration of between 2-20% (w/w) in a composition comprising a plurality of active substances or metabolites, all of which contribute to a synergistic functional effect and thus may significantly reduce the concentration in use, as well as reduce side effects and any toxicity or irritation that may be observed when these compounds are used at high concentrations >2% (w/w).

The present invention relates to compositions comprising active substances produced by the growth of probiotic microorganisms. The composition may be administered for internal or external use. External use may be applied topically as a living microorganism, for example to the scalp, to the feet and/or to the nails, or as a metacomposition comprising the active substance.

The present invention may relate to compositions comprising anti-dandruff actives produced by the growth of probiotic microorganisms and applied topically to the scalp as viable microorganisms or as a metacomposition comprising anti-dandruff actives.

In an embodiment of the invention, the composition comprises a probiotic microorganism capable of producing bacteriocins and at least one organic acid as active substance.

In another embodiment of the invention, the composition comprises a probiotic microorganism capable of producing a plantaricin and at least one organic acid as active substances.

Preferably, the composition comprises at least 2 different bacteriocins as active substances.

The compositions of the present invention may comprise probiotic microorganisms capable of producing an active substance.

The compositions of the present invention may include an acceptable carrier, which for topical compositions may be a dermatologically acceptable carrier (which may be referred to as a "carrier") for providing the active agent. Suitable carriers may be selected to produce the desired product form. In addition, the solubility or dispersibility of the components can determine the form and nature of the carrier. In one embodiment, the carrier may be present at the following levels: about 30 wt% to about 99 wt%, about 40wt% to about 98 wt%, about 50wt% to about 96 wt%, or alternatively about 60 wt% to about 95 wt%, by weight of the composition. Wt% is based on the weight of the overall composition.

The carrier may take a variety of forms. Non-limiting examples include simple solutions (e.g., based on aqueous, organic solvents, or oils), emulsions, and solid forms (e.g., gels, powders, sticks, flowable solids, or amorphous materials). In certain embodiments, the carrier is an aqueous carrier, which may include water or natural plant juice, such as aloe vera water. In certain embodiments, the carrier may be in the form of an emulsion. Emulsions can generally be divided into having a continuous aqueous phase (e.g., oil-in-water and water-in-oil-in-water) or a continuous oil phase (e.g., water-in-oil and oil-in-water-in-oil). The oil phase of the present invention may include natural oils, vegetable oils, silicone oils, non-silicone oils (such as hydrocarbon oils), esters, ethers, and the like, and mixtures thereof.

For example, for emulsions, the aqueous phase may include water, such as demineralised water or distilled water. Other acceptable carriers that can be used in the aqueous carrier include, but are not limited to, alcohols or ether compounds such as ethanol, glycerol, dipropylene glycol, propylene glycol, butylene glycol, 1, 4-butylene glycol, 3-allyloxy-1, 2-propanediol, dipropylene glycol n-butyl ether, 1, 2-hexanediol, isosorbide dimethyl ether, ethanol, 1, 3-butanediol, 1, 3-propanediol, 2' -thiodiethylene glycol, and 1, 6-hexanediol, or combinations thereof.

In one embodiment of the invention, the carrier and active substance may be divided into two compartments, and the contents of the two compartments are then mixed into one composition prior to administration (such as by topical administration).

For example, a bottle comprising a carrier and a cap having a compartment comprising lyophilized live probiotic microorganisms, which may be released from the cap compartment prior to administration.

The composition according to the invention may have a pH in the range of about 3.0 to about 6.5, which may be measured by direct pH measurement using a standard hydrogen electrode of the composition at 25 degrees celsius. Thus, the pH of the composition may be in the range of about 3 to about 6, more preferably in the range of 3.5 to 5.5.

Preferred embodiments of the present invention are compositions, such as anti-dandruff compositions, having a pH of less than 6.5, more preferably a pH of less than 6, more preferably a pH of less than 5.5, and more preferably a pH of less than 5.2.

In embodiments of the invention, the compositions may be formulated as soaps, shampoos, emulsions; an oil; a paste; a powder; talc; a lotion; foaming; gel; an ointment; a suspension; a mist (mist); or a liquid; or a tablet.

In another embodiment of the invention, the compositions of the invention may be prepared as typical formulations for topical application. They may preferably be in the form: solutions, dispersions, emulsions, powders, talc, capsules, spheres, sponges, solid dosage forms, foams and other delivery mechanisms.

The compositions of embodiments of the present invention may be hair tonic, leave-on hair products such as scalp serum, conditioning agents, therapeutic agents and styling products, rinse-off hair products such as conditioning agents, shampoos and therapeutic agent products; and any other form that can be applied to the scalp or skin.

The compositions of the present invention may be leave-on compositions or rinse-off compositions, depending on the intended use.

In a preferred embodiment of the invention, the composition may be an anti-dandruff composition, and is preferably a leave-on scalp therapeutic agent.

In another preferred embodiment of the invention, the composition may be a shampoo.

In another embodiment of the invention, the composition may be a nail composition. In addition to the description of the composition formulation above, the nail composition may be nail polish (nail polish), nail cream, nail polish, nail lotion, nail gel, ointment, and the like.

The nail composition may be suitable for removing, limiting, treating or preventing a fungal infection, such as mycosis or fungal nail disease, on the nail of a human or animal.

In yet another embodiment of the present invention, the composition may be a foot composition. In addition to the description of the formulation of the composition above, the foot composition may preferably be a foot cream, foot oil, foot lotion, foot ointment, or the like.

The foot composition may be suitable for removing, limiting, treating or preventing fungal infections, such as mycoses or mycoses of the foot, tinea pedis, on the foot of a human or animal.

Other ingredients of the composition

Thus, the composition according to the invention may also comprise other usual hair ingredients. CTFA Cosmetic ingredients manual (CTFA Cosmetic Ingredient Handbook), tenth edition published by the company of the Association of cosmetics, toiletries and perfumes, washroom, AND FRAGRANCE Association, inc (now known as the personal care products committee (Personal Care Products Council)), 2004, describes a wide variety of non-limiting substances that can be added to the compositions herein. Examples of such component categories include, but are not limited to: abrasives, absorbents, fragrances, pigments, colorants/colorants, essential oils, skin-feel agents (sensate), astringents, anti-acne agents, anti-caking agents, defoamers, antimicrobial agents, antioxidants, binders, biological additives, buffers, leavening agents, chelating agents, film formers, opacifying agents, pH adjusting agents, propellants, reducing agents, sequestering agents, rheology modifiers, conditioning agents, emulsifiers, and surfactants. According to embodiments, the anti-dandruff composition may be formulated as a hair care composition, such as a shampoo, a hair conditioner, or a shampoo-conditioner combination, further comprising one or more of the following ingredients; (i) surfactants (anionic, amphoteric/zwitterionic, nonionic), (ii) conditioning agents, (iii) emulsifiers, (iv) opacifiers, (v) thickeners and (vi) buffers.

Surface active agent

Thus, in one embodiment, the compositions of the present invention may be formulated as hair care compositions having a shampoo base comprising at least one detersive surfactant selected from the group consisting of: anionic surfactant, nonionic surfactant, amphoteric surfactant, or combinations thereof.

The hair care compositions of the present invention may include detersive surfactants which provide cleaning performance to the composition.

The concentration of detersive surfactant component in the composition, such as a hair care composition, should be sufficient to provide the desired cleaning and lathering performance, and generally ranges from about 2 wt% to about 50 wt%, from about 5 wt% to about 40 wt%, from about 8 wt% to about 35 wt%, or from about 10 wt% to about 30 wt%. Thus, a composition such as a hair care composition may for example comprise the following amount of detersive surfactant: about 5 wt%, about 10 wt%, about 12 wt%, about 15 wt%, about 17 wt%, about 18 wt%, about 20 wt%, about 25 wt%, about 30 wt%, or a range between any two of the foregoing. Wt% is based on the weight of the overall composition.

Exemplary anionic surfactants for use in the compositions may include: ammonium lauryl sulfate, ammonium lauryl ether sulfate, triethylamine lauryl ether sulfate, triethanolamine lauryl ether sulfate, monoethanolamine lauryl ether sulfate, diethanolamine lauryl ether sulfate, sodium laurate, sodium lauryl sulfate, sodium lauryl ether sulfate, potassium lauryl ether sulfate, sodium lauryl sarcosinate, sodium lauroyl sarcosinate, lauryl sarcosinate, cocoyl sarcosinate, ammonium cocoyl sulfate, ammonium lauroyl sulfate, sodium cocoyl sulfate, sodium lauroyl sulfate, potassium cocoyl sulfate, potassium lauryl sulfate, triethanolamine lauryl sulfate, monoethanolamine cocoyl sulfate, monoethanolamine lauryl sulfate, sodium tridecyl benzene sulfonate, sodium dodecyl benzene sulfonate, sodium cocoyl hydroxyethyl sulfonate, and combinations thereof. In another embodiment of the invention, the anionic surfactant is sodium lauryl sulfate, sodium laureth sulfate, or a combination thereof.

Amphoteric/zwitterionic surfactants suitable for use in the compositions herein can include those surfactants which are known for use in, for example, hair care or other personal care cleaning. The concentration of such surfactants may range from about 0.5 wt% to about 20 wt% and from about 1 wt% to about 10 wt%. Examples are betaines, and further non-limiting examples of suitable zwitterionic or amphoteric surfactants are described in U.S. Pat. nos. 5,104,646 and 5,106,609, which are incorporated herein by reference in their entirety.

Amphoteric detersive surfactants suitable for use in the composition can include those surfactants which are broadly described as derivatives of aliphatic secondary and tertiary amines in which the aliphatic radical can be straight or branched chain and wherein one of the aliphatic substituents contains from about 8 to about 18 carbon atoms and contains at least one anionic group such as carboxy, sulfonate, sulfate, phosphate, or phosphonate. Zwitterionic detersive surfactants suitable for use in the hair care composition include those surfactants which are broadly described as derivatives of aliphatic quaternary ammonium, phosphonium, and sulfonium compounds, in which the aliphatic radicals can be straight chain or branched, and wherein one of the aliphatic substituents contains from about 8 to about 18 carbon atoms and one contains an anionic group, such as carboxy, sulfonate, sulfate, phosphate, or phosphonate. Exemplary amphoteric and/or zwitterionic detersive surfactants for use in the compositions of the present invention include cocoyl amphoacetate, cocoyl amphodiacetate, lauroyl amphoacetate (lauroamphoacetate), lauroyl amphodiacetate (lauroamphodi acetate), cocamidopropyl betaine, cocamidopropyl hydroxysulfobetaine, and mixtures thereof.

Nonionic surfactants can be added to some compositions, particularly shampoo compositions.

Compositions according to the invention, such as shampoo compositions, may include nonionic surfactants. Nonionic surfactants can include those compounds produced by condensing alkylene oxide groups that are hydrophilic in nature with an organic hydrophobic compound that can be aliphatic or alkyl aromatic in nature.

Non-limiting examples of nonionic surfactants, such as for shampoo compositions, can include the following: (1) Polyethylene oxide condensates of alkylphenols, such as the condensation products of alkylphenols having alkyl groups containing from about 6 to about 20 carbon atoms in a straight or branched chain configuration, having ethylene oxide present in an amount equivalent to from about 10 to about 60 moles of ethylene oxide per mole of alkylphenol; (2) Those derived from the condensation of ethylene oxide with the product resulting from the reaction of propylene oxide and ethylenediamine; (3) Condensation products of aliphatic alcohols having from about 8 to about 18 carbon atoms in a straight or branched configuration with ethylene oxide, such as coconut alcohol ethylene oxide condensates having from about 10 to about 30 moles of ethylene oxide per mole of coconut alcohol, the coconut alcohol fraction having from about 10 to about 14 carbon atoms; (4) A long chain tertiary amine oxide of formula [ R ¹ R²R³ N0 ], wherein R ¹ comprises an alkyl, alkenyl or monohydroxyalkyl group of from about 8 to about 18 carbon atoms, from 0 to about 10 ethylene oxide moieties, and from 0 to about 1 glyceryl moiety, and R ² and R ³ comprise from about 1 to about 3 carbon atoms and from 0 to about 1 hydroxyl group, such as methyl, ethyl, propyl, hydroxyethyl or hydroxypropyl groups; (5) Long chain tertiary phosphine oxides of the formula [ RR 'R "P (R) 0], wherein R comprises an alkyl, alkenyl or monohydroxyalkyl group having a chain length ranging from about 8 to about 18 carbon atoms, 0 to about 10 ethylene oxide moieties, and 0 to 1 glyceryl moiety, and R' and R" are each an alkyl or monohydroxyalkyl group comprising from about 1 to about 3 carbon atoms; (6) A long chain dialkyl sulfoxide comprising one short chain alkyl or hydroxyalkyl group (typically methyl) of 1 to about 3 carbon atoms and one long hydrophobic chain comprising an alkyl, alkenyl, hydroxyalkyl or ketoalkyl group comprising about 8 to about 20 carbon atoms, 0 to about 10 ethylene oxide moieties, and 0 to 1 glyceryl moiety; (7) An Alkyl Polysaccharide (APS) surfactant (e.g., an alkyl polyglycoside), examples of which are described in U.S. Pat. No. 4,565,647, the entire contents of which are incorporated herein by reference, and which disclose APS surfactants having a hydrophobic group of about 6 to about 30 carbon atoms and a polysaccharide (e.g., a glycoside) as a hydrophilic group; alternatively, there may be a polyalkylene oxide group linking the hydrophobic moiety and the hydrophilic moiety; and alkyl groups (i.e., hydrophobic moieties) may be saturated or unsaturated, branched or unbranched, and unsubstituted or substituted (e.g., substituted with hydroxy groups or rings); and (8) polyoxyethylene alkyl ethers of the formula RO (CH ₂CH2)_n H), and polyethylene glycol (PEG) glycerol fatty acid esters of the formula R (0) 0CH ₂CH(0H)CH₂(0CH₂CH₂)_n H, wherein n is from 1 to about 200, preferably from about 20 to about 100, and R is an alkyl group having from about 8 to about 22 carbon atoms.

Certain nonionic surfactants may also be used as foam stabilizers, viscosity control agents, or conditioning agents. When included, the composition may comprise from about 0.5 wt% to about 5.0 wt% nonionic surfactant, or from about 0.75 wt% to about 2.0 wt%. Non-limiting examples of other anionic, amphoteric/zwitterionic, nonionic, or optional additional surfactants suitable for use in the compositions are described in U.S. Pat. nos. 3,929,678, 2,658,072;2,438,091;2,528,378, which is incorporated herein by reference in its entirety.

Conditioning agent

In one embodiment of the present invention, the composition may include one or more conditioning agents. Conditioning agents include substances used to impart specific conditioning benefits to human or animal hair, nails, and/or skin. Conditioning agents useful in the compositions of the present invention generally include water insoluble, water dispersible, nonvolatile liquids that form emulsified liquid particles. Suitable conditioning agents for use in the composition may be those generally characterized as follows: polysiloxanes (e.g., silicone oils, cationic polysiloxanes, silicone gums, high refractive polysiloxanes, and silicone resins), organic conditioning oils (e.g., hydrocarbon oils, polyolefins, and fatty esters), or combinations thereof, or those conditioning agents that otherwise form liquid, dispersed particles in an aqueous surfactant matrix. In embodiments, the one or more conditioning agents are present at about 0.01 wt% to about 10 wt%, about 0.1 wt% to about 8 wt%, and about 0.2 wt% to about 4 wt% by weight of the overall composition.

Emulsifying agent

As described below, a variety of anionic emulsifiers may be used in the compositions of the present invention, particularly shampoo compositions. By way of illustration, anionic emulsifiers include, but are not limited to, alkyl sulfates, alkyl ether sulfates, alkyl isothionates, alkyl carboxylates, alkyl sulfosuccinates, alkyl succinates, alkyl sulfates such as sodium dodecyl sulfate, water soluble salts of alkyl sarcosinates, alkyl derivatives of protein hydrolysates, acyl aspartate, alkyl or alkyl ether or alkylaryl ether phosphates, sodium dodecyl sulfate, phospholipids or lecithins, or soaps, sodium, potassium or ammonium stearate, oleates or palmitates, alkylaryl sulfonates such as sodium dodecyl benzene sulfonate, sodium dialkyl sulfosuccinate, sodium dioctyl sulfosuccinate, sodium dilauryl sulfosuccinate, sodium poly (styrene sulfonate) sodium salt, isobutylene-maleic anhydride copolymers, gum arabic, sodium alginate, carboxymethyl cellulose, cellulose sulfate and pectin, poly (styrene sulfonate), isobutylene-maleic anhydride copolymers, gum arabic, carrageenan, sodium alginate, pectic acid, astragalus, almond gum and agar; semisynthetic polymers such as carboxymethyl cellulose, sulfated methyl cellulose, carboxymethyl starch, phosphorylated starch, lignin sulfonic acid; and synthetic polymers such as maleic anhydride copolymers (including hydrolysis products thereof), polyacrylic acid, polymethacrylic acid, butyl acrylate copolymers or crotonic acid homopolymers and copolymers, vinylbenzenesulfonic acid or 2-acrylamido-2-methylpropanesulfonic acid homopolymers and copolymers, as well as partial amides or partial esters of such polymers, carboxy-modified polyvinyl alcohol, sulfonic acid-modified polyvinyl alcohol, and phosphoric acid-modified polyvinyl alcohol, phosphorylated or sulfated tristyrylphenol polyoxyethylene ether.

In addition, anionic emulsifiers having acrylate functionality may also be used in the compositions of the present invention, such as the shampoo compositions of the present invention. Anionic emulsifiers useful herein include, but are not limited to: poly (meth) acrylic acid; copolymers of (meth) acrylic acid and (meth) acrylic esters with C1-22 alkyl, C1-C8 alkyl, butyl; copolymers of (meth) acrylic acid and (meth) acrylamide; a carboxyvinyl polymer; acrylate copolymers such as acrylate/C10-30 alkyl acrylate crosslinked polymer, acrylic acid/vinyl ester copolymer/acrylic acid/vinyl isodecanoate crosslinked polymer, acrylate/palm oleyl polyether-25 acrylate copolymer, acrylate/stearyl alcohol polyether-20 itaconate copolymer, and acrylate/celeth-20 itaconate copolymer; polystyrene sulfonate, copolymers of methacrylic acid and acrylamide methylpropanesulfonic acid, and copolymers of acrylic acid and acrylamide methylpropanesulfonic acid; carboxymethyl cellulose; carboxyl guar gum; copolymers of ethylene with maleic acid; and (3) an acrylate silicone polymer. In embodiments, the emulsifier, when present, ranges from about 0.01 weight percent to about 5 weight percent, or from about 0.1 weight percent to about 4 weight percent, or from about 0.1 weight percent to about 3 weight percent, based on the weight of the overall composition. Wt% is based on the weight of the overall composition.

Carbomers can be used for hydrogels, low concentrations for liquid gels, and higher concentrations for solid gels.

Optional opacifying agent

Some compositions of the present invention may be provided in opaque formulations by incorporating substances therein to achieve a cosmetically attractive pearl-like appearance, known as pearlescence. The opacifying or pearlescent material may include, but is not limited to, titanium dioxide coated mica, iron oxide coated mica, ethylene glycol monostearate, ethylene glycol distearate, polyethylene glycol distearate, bismuth oxychloride coated mica, myristyl myristate, guanine, sparkling powder (polyester or metal), and mixtures thereof. Other pearlescent materials may be found in U.S. patent No. 4,654,207 and U.S. patent No. 5,019,376, which are incorporated herein by reference. In embodiments, the concentration of the opacifying agent, when present, ranges from about 0.01 weight percent to about 5 weight percent, or from about 0.1 weight percent to about 3 weight percent, or from about 0.1 weight percent to about 2 weight percent, by total weight. Wt% is based on the weight of the overall composition.

Thickening agent

Thickeners or rheology modifiers include, but are not limited to, acrylamide/ammonium acrylate copolymer (and) polyisobutylene (and) polysorbate 20; acrylamide/sodium acryloyldimethyl taurate copolymer/isohexadecane/polysorbate 80; an acrylate copolymer; acrylate/behenyl alcohol polyether-25 methacrylate copolymer; acrylate/C10-C30 alkyl acrylate crosslinked polymers; acrylate/steareth-20 itaconate copolymer; ammonium polyacrylate/isohexadecane/PEG-40 castor oil; cl 2-16 alkyl PEG-2 hydroxypropyl hydroxyethyl ethyl cellulose (HM-EHEC); carbomers; crosslinked polyvinylpyrrolidone (PVP); dibenzylidene sorbitol; hydroxyethyl Ethyl Cellulose (EHEC); hydroxypropyl methylcellulose (HPMC); hydroxypropyl methylcellulose (HPMC); hydroxypropyl cellulose (HPC); methylcellulose (MC); methyl hydroxyethyl cellulose (MEHEC); PEG-150/decanol/SMDI copolymer; PEG-150/stearyl alcohol/SMDI copolymer; polyacrylamide/C13-14 isoparaffin/laurinol polyoxyethylene ether-7; polyacrylate 13/polyisobutylene/polysorbate 20; polyacrylate crosslinked polymer-6; polyamide-3; polyquaternium-37 (and) hydrogenated polydecene (and) tridecyl alcohol polyether-6; polyurethane-39; sodium acrylate/acryloyldimethyl taurate/dimethylacrylamide; cross-linked polymer (and) isohexadecane (and) polysorbate 60; sodium polyacrylate and combinations thereof. In embodiments, the rheology modifier, when present, is present in a concentration ranging from about 0.01 wt% to about 7 wt%, or from about 0.1 wt% to about 5 wt%, or from about 0.2 wt% to about 4 wt%, by weight of the overall composition. Wt% is based on the weight of the overall composition.

Buffer solution

The compositions according to the present invention may have a pH in the range of about 3.0 to about 6.5, which may be stabilized by the presence of a buffer system. Suitable buffer solutions may be prepared using weak acid or weak base systems using, for example, citric acid, phosphoric acid, phthalic acid, acetic acid, lactic acid, glycine, or mixtures thereof. In each case, the proper buffering capacity is obtained by adjusting the final pH of the composition to within the above pH range. This can be accomplished using an acid (e.g., HCl, citric acid) or a base (e.g., naOH, sodium citrate) as desired. The amount of buffer used in the compositions of the present invention may depend on the particular acid selected, but is generally from about 0.1 wt% to about 10 wt%, preferably from about 0.2 wt% to about 5 wt%. Wt% is based on the weight of the overall composition.

Additional agents, such as benefit agents, may also be included in the compositions according to the present invention. The benefit agent may comprise a substance selected from the group consisting of: a prebiotic; a perfume; a brightening agent; an enzyme; sensates (cold or hot); an attractant, preservative; a dye; a pigment; a bleaching agent; and mixtures thereof.

It is further understood that the natural active combinations of the invention disclosed herein may also be used in combination with secondary benefit actives (such as secondary scalp benefit actives, e.g., soluble and/or insoluble secondary actives). Such secondary actives may include, but are not limited to, azoles such as ketoconazole, econazole (econazole), climbazole (climbazole), and neoconazole (elubiol); keratolytic agents such as salicylic acid; and zinc-containing layered (ZLM) materials, pyrithione anti-dandruff particles such as zinc pyrithione, coal tar, sulfur, activated carbon, huai Feier d (whitfield's) ointments, castelani (castellani's) paints, aluminum chloride, gentian violet, pyridone ethanolamine salts (piroctone olamine salts), ciclopirox olamine, undecylenic acid and metal salts thereof, potassium permanganate, selenium sulfide, sodium thiosulfate, propylene glycol, urea formulations, griseofulvin, 8-hydroxyquinoline ciloquinoline, thiobenzene, thiocarbamate, haloprogin, polyenes, hydroxypyridones, morpholine, benzylamine, allylamine (such as terbinafine), SENSIVA SC-50, elestab HP-100, azelaic acid, lysozyme, iodopropynyl butylcarbamate (IPBC), isothiazolinones such as octyl isothiazolinone, other natural oils, extracts or compounds such as bitter orange oil, tea tree oil, clove leaf oil, coriander, horse clove, red, cinnamon oil, citronellal, citronellol, 34, and combinations thereof. In embodiments, the concentration of the secondary benefit agent may range from about 0.01 wt% to about 5 wt%, or from about 0.1 wt% to about 3 wt%, or from about 0.1 wt% to about 2 wt%, by weight of the overall composition. Wt% is based on the weight of the overall composition. In alternative embodiments of the invention, the natural active substance combinations of the invention disclosed herein may also be free of any of the aforementioned secondary active substances.

Preferably, at least one prebiotic compound may be included as an additional agent in the compositions of the present invention, i.e. as a further ingredient. From a very broad concept, prebiotics are all those compounds that can be metabolized by probiotics. Preferably, the prebiotic is indigestible or indigestible by the mammal. Prebiotics are well known in the art and, when used in the present invention, there are no particular restrictions on the prebiotic itself.

Preferably, the at least one prebiotic product in the composition may be selected from the following compounds and compositions: non-digestible carbohydrates, beta-glucan, mannooligosaccharides, inulin, fructooligosaccharides, human Milk Oligosaccharides (HMO), galactooligosaccharides (GOS), lactulose oligosaccharides, fructooligosaccharides (FOS), cellobiose, cellodextrins, cyclodextrins, maltitol, lactitol, glucosyl sucrose. Mannooligosaccharides and/or inulin may be preferred.

HMOs may include lacto-N-tetraose, lacto-N-fucose, lacto-N-trisaccharide, 3 '-sialyllactose, lacto-N-neofucopyranose, sialic acid, L-fucose, 2-fucosyllactose, 6' -sialyllactose, lacto-N-neotetraose and 3-fucosyllactose.

D-and L-fucose may be suitable and may enhance the natural defenses of the skin, stimulate the epidermal immune defenses and/or prevent and/or treat autoimmune diseases of the skin. In a preferred embodiment of the invention, the composition comprises D-and/or L-fucose.

In a preferred embodiment of the invention, the composition further comprises L-fucose in a concentration of 10mM to 500mM in the composition.

The compositions according to the present invention may generally be prepared by conventional methods known in the art, for example, methods of preparing shampoo compositions, creams, oils, emulsions, and the like. Such methods typically involve mixing the ingredients to a relatively uniform state in one or more steps, with or without the use of heat, cooling, application of vacuum, and the like. Compositions may be prepared such as to optimize stability (physical stability, chemical stability, photostability) and/or delivery of the active material. Such optimization may include appropriate pH, as well as excluding materials that may complex or react with one or more active agents and thus negatively impact stability or delivery.

The composition may be a single phase or a single product, or the composition may be a separate phase or a separate product. If two products are used, the products may be used together, simultaneously or sequentially. Sequential use may occur over a short period of time, such as immediately after use of one product, or over hours or days.

In one embodiment of the invention, the composition is a leave-on composition comprising viable probiotics at a concentration of 10 ³ to 10 ¹³ colony forming units per gram. More specifically, the composition comprises from 10 ⁴ to 10 ¹⁰ colony forming units of bacteria per gram, more specifically, the composition comprises from 10 ⁵ to 10 ⁹ colony forming units of bacteria per gram.

In one embodiment of the invention, the composition is a leave-on composition comprising viable probiotics of bacteria at a concentration of at least 10 ³ colony forming units per gram. More specifically, the composition comprises at least 10 ⁴ colony forming units of bacteria per gram, more specifically, the composition comprises at least 10 ⁵ colony forming units of bacteria per gram, more specifically, the composition comprises at least 10 ⁶ colony forming units of bacteria per gram, more specifically, the composition comprises at least 10 ⁷ colony forming units of bacteria per gram.

In one embodiment of the invention, the composition comprises a metagenic fraction of an active, such as an anti-dandruff active or an antifungal active.

Use of the composition according to the invention as an anti-dandruff composition: according to an embodiment of the present invention, there is provided a method for treating a subject suffering from dandruff and/or preventing or inhibiting the onset of dandruff symptoms associated with the proliferation of malassezia yeasts on the scalp of a subject. The method comprises contacting the scalp or keratinous tissue of a subject with an effective amount of an anti-dandruff composition of the invention. The anti-dandruff composition may be massaged onto the scalp and should remain in contact with the scalp or keratinous tissue of the subject for at least 15 seconds or more. Depending on the formulation, the anti-dandruff composition may be leave-on, or it may be rinsed off.

Thus, in another embodiment, the method comprises topically applying an anti-dandruff composition comprising an effective amount of an anti-dandruff active to an area of skin of a subject in need or desiring to inhibit malassezia, wherein the anti-dandruff active remains in contact with the area as a leave-on composition or for 15 seconds or more; and then rinsed off. In yet another embodiment, the method comprises administering the composition according to a rule, wherein the rule comprises: (a) cleansing the scalp to form a cleansed scalp; (b) An anti-dandruff composition is topically applied to the cleaned scalp.

Use of the composition according to the invention as an antifungal composition: according to an embodiment of the present invention, there is provided a method for treating a subject suffering from nail or foot mycoses and/or preventing or inhibiting the onset of mycosis symptoms associated with malassezia proliferation on the nail and/or foot of a subject. The method comprises contacting the nail and/or foot of a subject with an effective amount of an antifungal composition of the present invention. The antifungal composition may be massaged onto the nails and/or feet and should remain in contact with the nails and/or feet of the subject for at least 15 seconds or more. Depending on the formulation, the antifungal composition may be leave-on or it may be rinsed off.

Thus, in another embodiment, the method comprises topically applying an antifungal composition comprising an effective amount of an antifungal active to an area of the nail and/or foot of a subject in need or desiring to inhibit fungi, wherein the antifungal active remains in contact with the area as a leave-on composition or for 15 seconds or more; and then rinsed off. In yet another embodiment, the method comprises administering the composition according to a rule, wherein the rule comprises: (a) cleansing the scalp to form a cleansed nail or foot; (b) Applying an antifungal composition topically to the cleaned nail or foot.

According to the invention, the fungal infection may include dandruff and/or mycosis. Mycoses may relate to any disease caused by fungi and may affect different parts of the body, in particular the feet, ears, hands and/or nails of the human or animal body.

Preferred embodiments of the present invention relate to the use of a composition according to the invention for the prevention and/or treatment of mycoses in humans or animals.

Preferably, preventing and/or treating mycosis and/or dandruff in a human or animal may comprise preventing and/or treating a condition associated with proliferation of malassezia yeast on the scalp, hands, ears, feet, nails, or skin of a subject.

In another embodiment of the invention, a composition comprising one or more probiotic strains (preferably isolated probiotic strains) and/or one or more active substances of one or more probiotic strains (preferably isolated probiotic strains) is used for the prevention and/or treatment of mycoses in humans or animals.

The prevention and/or treatment of mycoses in humans or animals may be prevention and/or treatment:

-dandruff conditions associated with the proliferation of malassezia yeasts on the scalp or skin of a human or animal; or alternatively

-Fungal nail infections associated with the proliferation of fungi or yeasts such as onychomycosis (Tinea unguium) or trichophyton (Trichophyton spp.) on human or animal nails; or alternatively

In an embodiment of the invention, the composition comprises nucleic acids and/or nucleotides.

In another embodiment of the invention, the composition does not comprise plant matter and/or fibrous matter. Preferred fibrous materials are plant fibrous materials

Preferably, the content of fibrous material in the composition is less than 5% (w/w), such as less than 4% (w/w), for example less than 3% (w/w), such as less than 2% (w/w), for example less than 1% (w/w), such as less than 0.5 (w/w), for example less than 0.1% (w/w), such as 0%, relative to the overall composition.

In an embodiment of the invention, the composition comprises two or more active substances (specific active substances may be as described above) of one or more isolated probiotic strains, such as 3 or more, for example 4 or more, such as 5 or more, for example 4 or more, such as 10 or more, for example 15 or more, such as 20 or more, for example 25 or more.

The subject according to the invention may be a mammal selected from the group consisting of human, dog, cat, horse, cow or sheep.

The composition according to the invention may be used daily, weekly or on a variety of rules. The composition may be used more than once per day, such as in the evening and morning. The composition may be used after washing hair (and may also be on wet or dry hair) or after washing the body, feet or nails, which may mean that the composition is used more than once a day or only a few times a week on some days. The composition may be used three times daily, twice daily, once daily, six times per week, five times per week, four times per week, three times per week, twice per week, or once per week. In some embodiments, the anti-dandruff composition is used four, five, six, or seven times per week.

According to another embodiment, the composition may be administered at least once a day for at least about a week, or at least twice a day for at least about a week.

According to another embodiment, the composition may be administered at least once daily for at least about four weeks, or at least twice daily for at least about four weeks. According to another embodiment, the composition may be administered at least once daily for at least about eight weeks. The composition can be used by both men and women. The composition can be used by mammals of any age, including newborns, infants, young children and children.

In embodiments of the invention, the composition may be an anti-dandruff composition. Preferably, the anti-dandruff composition is useful for preventing or treating eschar.

In another embodiment of the invention, the composition may be applied to the skin, fur, oral cavity, genital area, ear and/or body canal of a human or animal.

Preferably, the composition according to the invention may be topically applied to an animal. In a further preferred embodiment, the animal may be selected from the group consisting of dogs, cats, cattle, cows, sheep, goats, horses and birds.

The present invention may relate to compositions (such as anti-dandruff compositions and/or antifungal compositions) comprising probiotic microorganisms capable of producing an active agent when topically applied to an affected area (such as scalp, hands, skin, ears, feet, and/or nails).

The present invention may relate to an anti-dandruff composition comprising probiotic microorganisms capable of producing anti-dandruff actives when topically applied to the scalp.

Preferably, the composition comprises at least 2 anti-dandruff actives produced by probiotic microorganisms selected from the group consisting of: bacteriocins, lactic acid, acetic acid, succinic acid, azelaic acid, salicylic acid, indole-3-lactic acid, indole-3-acetic acid, 2-hydroxybutyric acid, N-acetyltryptophan, glycolic acid, N-acetylglutamine and N-acetylaspartic acid.

A composition for use in the treatment of dandruff and/or mycosis, wherein the composition comprises viable probiotic microorganisms at a concentration of at least 10 ³ colony forming units per gram of the composition, wherein the probiotic microorganisms are capable of producing an anti-dandruff and/or antifungal active.

A composition having a pH below 6.5 for use in the treatment of dandruff and/or mycosis, wherein the composition comprises viable probiotic microorganisms at a concentration of at least 10 ³ colony forming units per gram of the composition, wherein the probiotic microorganisms are capable of producing an anti-dandruff and/or antifungal active.

A composition comprising a bacteriocin-producing probiotic microorganism, wherein the same microorganism produces at least two of the following metabolites: succinic acid, azelaic acid, salicylic acid, 2-hydroxyisocaproic acid, indole-3-lactic acid, indole-3-acetic acid, 2-hydroxybutyric acid, N-acetyltryptophan, glycolic acid, N-acetylglutamine and N-acetylaspartic acid.

A composition comprising a metagenic fraction from lactic acid bacteria capable of producing at least two of the following anti-dandruff and/or antifungal actives: succinic acid, azelaic acid, salicylic acid, 2-hydroxyisocaproic acid, indole-3-lactic acid, indole-3-acetic acid, 2-hydroxybutyric acid, N-acetyltryptophan, glycolic acid, N-acetylglutamine or N-acetylaspartic acid.

An anti-dandruff and/or anti-fungal composition comprising indole-3-lactic acid wherein the indole-3-lactic acid is produced by lactic acid bacteria.

An anti-dandruff and/or antifungal composition comprising salicylic acid, wherein the salicylic acid is produced by lactic acid bacteria.

An anti-dandruff and/or anti-fungal composition comprising gluconic acid, wherein the gluconic acid is produced by lactic acid bacteria.

An anti-dandruff and/or anti-fungal composition comprising phenyllactic acid, wherein the phenyllactic acid is produced by lactic acid bacteria.

An anti-dandruff and/or antifungal composition comprising 2-hydroxyisocaproic acid, wherein the 2-hydroxyisocaproic acid is produced by lactic acid bacteria.

An anti-dandruff and/or antifungal composition comprising 2-hydroxy-butyric acid, wherein 2-hydroxy-butyric acid is produced by lactic acid bacteria.

An anti-dandruff and/or antifungal composition comprising N-acetyl aspartic acid, wherein the N-acetyl aspartic acid is produced by lactic acid bacteria.

An anti-dandruff and/or anti-fungal composition comprising azelaic acid wherein azelaic acid is produced by lactic acid bacteria.

An anti-dandruff and/or anti-fungal composition comprising succinic acid, wherein succinic acid is produced by lactic acid bacteria.

An anti-dandruff and/or antifungal composition comprising indole 3-acetic acid, wherein indole 3-acetic acid is produced by lactic acid bacteria.

An anti-dandruff and/or antifungal composition comprising 3-phenyllactic acid, wherein the 3-phenyllactic acid is produced by lactic acid bacteria.

An anti-dandruff and/or antifungal composition comprising N-acetylglutamine, wherein the N-acetylglutamine is produced by lactic acid bacteria.

An anti-dandruff and/or antifungal composition comprising 2-hydroxyisocaproic acid and salicylic acid produced by lactic acid bacteria.

An anti-dandruff and/or anti-fungal disease composition according to any preceding composition wherein the composition comprises at least one bacteriocin.

An anti-dandruff and/or antifungal composition comprising bacteriocin, salicylic acid, indole-3-lactic acid and 3-phenyllactic acid.

Anti-dandruff and/or antifungal composition comprises bacteriocin, 2-hydroxyisocaproic acid, salicylic acid, indole-3-lactic acid, 2-hydroxybutyric acid and N-acetyl aspartic acid

An anti-dandruff and/or anti-fungal composition according to any of the above compositions wherein the composition is produced from isolated lactic acid bacteria.

An anti-dandruff and/or anti-fungal composition according to any of the above compositions, further comprising at least one antimicrobial bacterial metabolite selected from the group consisting of: hydrogen peroxide, 3-phenyllactic acid, 3-hydroxyphenyllactic acid, 4-hydroxyphenyllactic acid, 2-hydroxyisocaproic acid, 3-hydroxypropionaldehyde, 1, 2-propanediol, 1, 3-propanediol, succinic acid, ethanol, acetic acid, carbonic acid, propionic acid, butyric acid, cyclic dipeptide, cyclic (L-Phe-L-Pro), cyclic (L P-trans-4-OH-L-Pro), 3- (R) -hydroxydecanoic acid, 3-hydroxy-5-cic dodecanoic acid, 3- (R) -hydroxydodecanoic acid, and 3- (R) -hydroxytetradecanoic acid.

An anti-dandruff and/or anti-fungal composition according to any of the above compositions wherein the probiotic microorganism is selected from the group consisting of; lactobacillus plantarum LB356R (DSM 33094), lactobacillus plantarum LB244R (DSM 32996), wegener viridis LB10G (DSM 32906), lactobacillus paracasei LB113R (DSM 32907), lactobacillus paracasei LB116R (DSM 32908), lactobacillus brevis LB152G (DSM 32995), lactobacillus paracasei LB28R (DSM 32994), enterococcus faecium LB276R (DSM 32997), leuconostoc mesenteroides LB349R (DSM 33093), lactobacillus plantarum LB316R (DSM 33091), lactobacillus plantarum LB312R (DSM 33098), pediococcus pentosaceus LB606R (DSM 33730), lactobacillus plantarum LB679R (DSM 33731), lactobacillus crispatus LB714R (DSM 33732), lactobacillus griseus LB905R (DSM 34094), lactobacillus crispatus LB 152R (DSM 34095), lactobacillus jensenii LB918R (DSM 34096), lactobacillus helveticus LB919R (DSM 34097), lactobacillus paracasei subspecies (DSM 34249) R (DSM 34249), lactobacillus pentosus 34250, or any combination thereof.

Preservation of biological materials

The lactic acid bacteria according to the present invention comprise in particular microorganisms selected from the group comprising:

-the following microorganisms were deposited at the german collection of microorganisms and cell cultures (German Collection for Microorganisms and Cell Cultures) at 5.5.2022: lactobacillus plantarum LB681R (DSM 34250) and lactobacillus paracasei subspecies paracasei LB555R (DSM 34249);

The following microorganisms were deposited at the German collection of microorganisms and cell cultures at 12.14.2020: lactobacillus crispatus LB714R (DSM 33732), pediococcus pentosaceus LB606R (DSM 33730) and/or lactobacillus plantarum LB679R (DSM 33731);

the following microorganisms were deposited at 2021, 11/25: lactobacillus crispatus LB919R (DSM 34097), lactobacillus crispatus LB912R (DSM 34095), lactobacillus gasseri LB905R (DSM 34094) and/or lactobacillus jensenii LB918R (DSM 34096);

The following microorganisms were deposited at the German collection of microorganisms and cell cultures at 10/4/2019: lactobacillus plantarum LB356R (DSM 33094), leuconostoc mesenteroides LB349R (DSM 33093), lactobacillus paracasei LB316R (DSM 33091) and/or lactobacillus plantarum LB312R (DSM 33098);

The following microorganisms were deposited at the German collection of microorganisms and cell cultures at 13/12.2018: lactobacillus plantarum LB244R (DSM 32996), enterococcus faecium LB276R (DSM 32997), lactobacillus brevis LB152G (DSM 32995) and/or lactobacillus paracasei LB28R (DSM 32994); and/or

The following microorganisms were deposited at the German collection of microorganisms and cell cultures at 28 of 8.2018: lactobacillus plantarum LB116R (DSM 32908), lactobacillus paracasei LB113R (DSM 32907) and/or weissella viridis LB10G (DSM 32906).

All patent and non-patent references cited in this application are incorporated herein by reference in their entirety.

The invention will now be described in more detail in the following non-limiting examples.

Examples

Example 1:

Pityrosporum ovale with CCUG number 59937 was delivered by the university of Goldburg culture Collection (Culture Collection University Gothenburg). The strain was isolated from a 15 year old girl with pityriasis versicolor. Malassezia furfur (M.fursur) 59937 was inoculated into potato dextrose broth (Sigma) and incubated for five days at 30 ℃. After incubation, cultures were streaked with cotton sticks onto potato dextrose agar (Sigma) plates to form lawn. On top of streaked agar plates 20. Mu.L of 48 hour cultured Lactobacillus plantarum were spotted (DSM 32996), lactobacillus plantarum/>(DSM 33094) and Pediococcus pentosaceus/>(DSM 33730). The plates were incubated for five days at 30 ℃. After incubation, the plates were visually inspected. Around all lactic acid bacteria spots, a zone of inhibition of +1cm in malassezia furfur was observed.

Example 2.

Different formulations

An anti-dandruff shampoo composition:

shampoo preparation A11

The components are as follows:

Water 65% (w/w)

Metagen (Lactobio Lactobacillus plantarum)Fermentation lysate) 15% (w/w) -containing a water content of 95-98% (w/w) (equivalent to a cell concentration of 10 ⁷-10¹⁰ cells per ml)

Lauroyl/myristoyl methyl glucamide 7% (w/w)

Decyl glucoside 2% (w/w)

Betaine 2% (w/w)

Sodium cocoyl isethionate 1% (w/w)

Glycerol 1% (w/w)

Coco-oleamide mipa% (w/w)

Hydrogenated coconut acid 1% (w/w)

Sorbic acid 1% (w/w)

Sodium isethionate 1% (w/w)

Sodium phytate 1% (w/w)

Ethanol 1% (w/w)

Sodium benzoate 0.5% (w/w)

Potassium sorbate 0.5% (w/w)

The pH was adjusted to 4.5

All ingredients except the probiotic micro-organisms were heated to melt, stirred to cool, and when the shampoo was cooled to 20 ℃, the probiotic micro-organisms were added as a metasolution.

Shampoo formulation a12:

The components are as follows:

Water 39% (w/w)

Metagen (Lactobacillus plantarum)Fermentation lysate) 14% (w/w) -containing a water content of 95-98% (w/w) (equivalent to a cell concentration of 10 ⁷-10¹⁰ cells per ml)

13% Sodium laureth sulfate (w/w)

Cocoyl amphodiacetate disodium 8% (w/w)

PEG-200 hydrogenated glyceryl palmitate 5% (w/w)

Polysorbate 20.5% (w/w)

Hexanediol 2% (w/w)

Castor oil amide disodium (Disodium Ricinoleamido) MEA 2% (w/w)

Sulfosuccinic acid 2% (w/w)

Sodium benzoate 1% (w/w)

Polysorbate 21 1% (w/w)

Sodium methylparaben 1% (w/w)

Polyquaternium-10% (w/w)

PEG-7 glyceryl cocoate, menthol 1% (w/w)

Ethyl parahydroxybenzoate 1% (w/w)

Propylene glycol 1% (w/w)

1% (W/w) of butylphenyl methylpropionaldehyde

Benzyl alcohol 1% (w/w)

Sodium hydroxide 1% (w/w)

The pH was adjusted to 3.7

Shampoo preparation A13

The components are as follows:

Water 21% (w/w)

20% Of sodium cocosulfate (w/w)

Coco glucoside 20% (w/w)

Glycerol 12% (w/w)

Sodium cocoyl isethionate 6% (w/w)

Lauryl glucoside 5% (w/w)

Stearyl citrate 4% (w/w)

Grape (VITIS VINIFERA) (grape)/raisin) seed oil 4% (w/w)

Citric acid 3% (w/w)

Lactobacillus plantarum3% (W/w) (lyophilized powder, concentration 9X10 ¹⁰ CFU/g)

Panthenol (vitamin B5) 0.5% (w/w)

Oleic acid glyceride 0.5% (w/w)

Inulin 0.5% (w/w)

Potassium sorbate 0.5% (w/w)

All ingredients except the probiotic micro-organisms were heated until melted, stirred to cool, the pH was adjusted to 5.0, and the probiotic micro-organisms were added when the shampoo was cooled to 20 ℃.

Shampoo preparation A14

The components are as follows:

40% water (w/w)

18% (W/w) of sodium C14-16 olefin sulfonate

Cocamidopropyl betaine 8% (w/w)

Sodium chloride 4% (w/w)

Cocoamidopropyl hydroxysulfobetaine 4% (w/w)

Coconut (Cocos Nucifera) (Coconut) oil 5%

PPG-2 hydroxyethyl coco/isostearamide 2%

Acrylate copolymer 2%

Inulin 2%

Lactobacillus plantarum2% (W/w) (lyophilized powder, concentration 2X10 ¹⁰ CFU/g)

Hydrolyzed milk protein 2%

Albumin 2%

Ammonia-terminated polydimethylsiloxane (Amodimethicone) 1%

Cocoyl isethionate sodium 1%

Glycol distearate 1%

Laureth-4 1%

Polyquaternium-6 1%

Polyquaternium-10%

Tridecyl alcohol polyether-12%

Qu Lvan 0.5% of Western medicine

Guar hydroxypropyl trimethyl ammonium chloride 0.5%

PEG-120 methyl glucose dioleate 0.5%

EDTA disodium 0.4%

Bis (hydroxymethyl) imidazolidinylurea 0.1%

The lyophilized probiotic microorganisms were encapsulated in a two-compartment plastic lid comprising 2g of lyophilized material (AccuRec kit two-phase lid, available from italian Bormioli Pharma).

All ingredients except the probiotic microorganisms were heated until melted, the pH was measured to 4.8, stirred cooled, and the 100ml bottles were filled with the composition and closed in separate compartments with a lid comprising the probiotic microorganisms. Before use, the microorganisms are released from the cap into the bottle and shaken well.

Dry shampoo powder; no-wash preparation B11

Corn (Zea Mays) starch 46% (w/w)

Aluminum starch octenyl succinate 38% (w/w)

Kaolinite 10% (w/w)

Cyclodextrin 5% (w/w)

Lactobacillus plantarum1% (W/w) (lyophilized powder, concentration 2X10 ¹⁰ CFU/g)

The lyophilized live probiotic microorganisms were mixed with cyclodextrin and subsequently mixed with the remaining ingredients at room temperature.

The anti-dandruff composition is sprayed or sprinkled onto the scalp as a leave-on composition.

Dry shampoo powder; no-wash preparation B12

Talc 97% (w/w)

Lactobacillus plantarum2% (W/w) (lyophilized powder, concentration 2X10 ¹⁰ CFU/g) inulin 1% (w/w)

The lyophilized live probiotic microorganisms were gently blended with talc at room temperature.

Dry shampoo powder; no-wash preparation B13

Corn starch 90% (w/w)

Talc 9% (w/w)

Pediococcus pentosaceus1% (W/w) (lyophilized powder, concentration 9x10 ⁹ CFU/g) lyophilized live probiotic microorganisms were gently blended with corn starch and talc at room temperature. The anti-dandruff composition is sprayed or sprinkled onto the scalp as a leave-on composition.

Dry shampoo powder; no-wash preparation B14

Potato starch 40% (w/w)

Corn starch 37% (w/w)

Kaolinite 10%8w/w

Talc 9% (w/w)

Cyclodextrin 4% (w/w)

Pediococcus pentosaceus1% (W/w) (lyophilized powder, at a concentration of 9x10 ⁹ CFU/g) the lyophilized live probiotic microorganisms were gently blended with the other ingredients at room temperature.

Shampoo leave-on preparation C11

Water 70% (w/w)

Apple vinegar 10% (w/w)

Cetrimide 5% (w/w)

Glycerol 3% (w/w)

Hydrolyzed vegetable protein 1% (w/w)

PG-propylsilanetriol 1% (w/w)

Aganshu kernel oil 1% (w/w)

Aloe vera leaf juice 1% (w/w)

Macadamia nut seed oil 1% (w/w)

Quaternary ammonium salt-96 1% (w/w)

Polyquaternium-55.1% (w/w)

Dipropylene glycol 1% (w/w)

Propylene glycol 1% (w/w)

Bis (hydroxymethyl) imidazolidinyl urea 1% (w/w)

Iodopropynyl butyl carbamate 1% (w/w)

Pediococcus pentosaceus1% (W/w) (lyophilized powder, concentration 9X10 ⁹ CFU/g)

All ingredients except the probiotic microorganisms were heated until melted, the pH was measured to 4.5, stirred cooled, and the 100ml bottles were filled with the composition and closed in separate compartments with a lid comprising the probiotic microorganisms. Before use, the microorganisms are released from the cap into the bottle and shaken well.

Shampoo no-wash preparation C12

Water 70% (w/w)

Apple vinegar (10% w/w)

Glycerol (3% w/w)

Rapeseed amidopropyl ethyl dimethyl ammonium ethyl sulfate 1% (w/w)

Polyacrylamide propyl trimethyl ammonium chloride 1% (w/w)

Hydrolyzed vegetable protein PG-propylsilanetriol 1% (w/w)

Aganshu kernel oil 1% (w/w)

Aloe vera leaf juice 1% (w/w)

Macadamia nut seed oil 1% (w/w)

Quaternary ammonium salt-96 1% (w/w)

Polyquaternium-55.1% (w/w)

Dipropylene glycol 1% (w/w)

Propylene glycol (Propanediol) 1% (w/w)

Propylene glycol (Propylene Glycol) 1% (w/w)

Bis (hydroxymethyl) imidazolidinyl urea 1% (w/w)

Tetrasodium glutamate diacetate 1% (w/w)

Iodopropynyl butyl carbamate 1% (w/w)

Pediococcus pentosaceus2% (W/w) (lyophilized powder, concentration 9X10 ⁹ CFU/g)

All ingredients except the probiotic micro-organisms were heated until melted and dissolved as a homogeneous solution, the pH was measured to 4.4, stirred cooled and 100ml bottles filled with the composition and closed in separate compartments with a lid comprising the probiotic micro-organisms. Before use, the microorganisms are released from the cap into the bottle and shaken well.

Shampoo no-wash preparation C13

Water 70% (w/w)

Lactobacillus plantarumFermentation lysate (15% w/w) -comprising a water content of 95-98% (w/w) (equivalent to a cell concentration of 10 ⁷-10¹⁰ cells per ml)

Glycerol (3% w/w)

Rapeseed amidopropyl ethyl dimethyl ammonium ethyl sulfate 1% (w/w)

Polyacrylamide propyl trimethyl ammonium chloride 1% (w/w)

Aganshu kernel oil 1% (w/w)

Aloe vera leaf juice 1% (w/w)

Macadamia nut seed oil 1% (w/w)

Quaternary ammonium salt-96 1% (w/w)

Polyquaternium-55.1% (w/w)

Dipropylene glycol 1% (w/w)

Propylene glycol 1% (w/w)

Bis (hydroxymethyl) imidazolidinyl urea 1% (w/w)

Tetrasodium glutamate diacetate 1% (w/w)

Gently heat until all ingredients are dissolved. Lactobacillus plantarum (L.plantarum) is added after cooling the composition to room temperatureAnd (3) fermenting the lysate. The pH was found to be 4.4.

Hair care essence leave-in preparation C14

Sunflower seed oil

Castor oil

Pediococcus pentosaceus LB606R 1% (w/w) (lyophilized powder, concentration 9X10 ⁹ CFU/g)

Tocopherol (vitamin E)

Hair care essence leave-in preparation C15

Sunflower seed oil 75% (w/w)

Castor oil 22% (w/w)

Tocopherol (vitamin E) (1% w/w)

Formulations C13 and C14 were mixed at room temperature without heating the ingredients.

The powder composition is useful not only as a dry shampoo, but also for scalp treatment, skin treatment, and also for diaper rash.

Topical leave-on formulation C16

Sunflower seed oil 55% (w/w)

Castor oil 42% (w/w)

Tocopherol (vitamin E) (1% w/w)

All ingredients were mixed at room temperature

Topical leave-on formulation C17

83.8% Water (w/w)

Pediococcus pentosaceusFermentation lysate (15% w/w) -comprising a water content of 95-98% (w/w) (equivalent to a cell concentration of 10 ⁷-10¹⁰ cells per ml)

Betaine 1% (w/w)

Carbomer 0.2% (w/w)

The water, betaine and carbomer were mixed until a clear solution was obtained, the pH was adjusted to pH 4.7 and the fermentation lysate was added.

Example 3

Metabonomics studies were performed in 3 different lactic acid bacteria. Lactobacillus plantarum LB356R (DSM 33094), lactobacillus plantarum LB244R (DSM 32996), pediococcus pentosaceus LB606R (DSM 33730).

Microorganisms were grown in MRS medium at 30℃for 24 hours. The supernatant was analyzed by semi-polar metabolite method. Sample analysis was performed by MS-OmicsDenmark) is performed as follows.

The samples were diluted 10-fold in 10mM ammonium formate containing 0.1% formic acid.

LC-MS method

Analysis was performed using a UPLC system (Vanquish, thermo FISHER SCIENTIFIC) in combination with a high resolution quadrupole Orbitrap mass spectrometer (Q Exactive ^TM HF Hybrid Quadrupole-Orbitrap, thermo FISHER SCIENTIFIC). The electrospray ionization interface serves as an ionization source. Analysis was performed in negative and positive ionization modes. QC samples were analyzed in MS/MS mode to identify compounds. UPLC is performed using a slightly modified version of the procedure described by Catalin et al (UPLC/MS Monitoring of Water-Soluble Vitamin Bs in Cell Culture Media in Minutes,Water Application note 2011,720004042en).

Data processing

Data were processed using Compound Discoverer 3.1 (ThermoFisher Scientific) and TRACEFINDER 4.1 (ThermoFisher Scientific).

Extraction of compounds

A compound will typically generate a signal in more than one mass trace (e.g., due to naturally occurring C13 isotopes, adducts, and/or fragments), and therefore a compound is almost always represented by more than one feature having the same retention time but different masses. The compound extraction performed by Compound Discoverer includes the following four steps:

1) First, features are extracted from the raw data.

2) Feature detection is then followed by feature grouping belonging to the same compound.

3) This additional information (e.g., isotopic form) is then used with the exact mass to determine the molecular formula.

4) The total information collected for each compound was then used in the following identification step.

Analysis was performed using Thermo Scientific Vanquish LC in combination with Thermo Q Exactive HF MS. The electrospray ionization interface serves as an ionization source. Analysis was performed in negative and positive ionization modes. UPLC is (UPLC/MS Monitoring of Water-Soluble Vitamin Bs in Cell Culture Media in Minutes,Water Application note 2011,720004042en). using Compound Discoverer 3.1 (Thermo Scientific) to extract peak area using a slightly modified version of the procedure described by Catalin et al. Identification of compounds was performed in four stages; stage 1: identification was performed by retention time (compared to internal true standard), exact mass (acceptable deviation of 3 ppm) and MS/MS spectra, grade 2 a: the identification was carried out by retention time (compared to internal authentic standards), exact quality (acceptable deviation of 3 ppm). Stage 2 b: identification was performed by accurate mass (3 ppm acceptable deviation) and MS/MS spectra, grade 3: the identification was performed only by accurate mass (acceptable deviation was 3 ppm).

1606 Compounds were co-detected in the samples. Of these, 271 kinds of 3-level notes, 103 kinds of 2 b-level notes, 113 kinds of 2 a-level notes, and 60 kinds of 1-level notes.

For the strains, lactic acid, acetic acid, succinic acid, azelaic acid, salicylic acid, indole-3-lactic acid, indole-3-acetic acid, 2-hydroxybutyric acid, 2-hydroxyisohexanoic acid and N-acetyl aspartic acid were all annotated in significant amounts on grade 1.

The concentration of the organic acid is greater than 3% (w/w) for all fermentations.

Example 4:

Dandruff evaluation test

Composition C14 was evaluated for 12 people with dandruff for 2 weeks.

The leave-on composition was massaged into the affected area of the scalp once a day for 14 days. The composition was allowed to stand for at least 8 hours prior to any washing.

The test subjects were asked to self-evaluate dandruff compared to dandruff prior to starting treatment. Dandruff was assessed according to the following score.

Relative to pre-treatment self-assessment	Score of
		Dandruff becomes more severe	-1
No change	0
		Significant reduction of dandruff	+1
Dandruff disappearing	+2

Results:

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All the testers observed a significant reduction in dandruff.

Example 5:

evaluation of treatment of malassezia infected skin in the outer ear of an irish soft beagle (dog age 11 years) by composition B12. The ear was gently washed with physiological saline, and powder composition B12 was sprayed into the ear, and the infected portion of the outer ear was gently massaged 2 times per day. After 2 days a significant therapeutic effect was observed, with redness in the ear, a mildewed odour and a reduced skin irritation. After 7 days, the treatment was reduced to once daily. After 2 weeks, the outer ear infection disappeared.

Example 6:

the effect of pH on the activity of the active substances was determined using the lawn spotting method described in example 1. Lactobacillus plantarum (DSM 32996), lactobacillus plantarum/>(DSM 33094) and Pediococcus pentosaceus/>(DSM 33730) each was grown in MRS medium at 30℃for 48 hours, the supernatant of each culture was separated by centrifugation and filtered to produce cell-free supernatant.

The supernatant was adjusted to the following pH with 1M NaOH and 1M HCl, respectively: 3. 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8. Phosphate buffer at each pH was used as a control for pH effects.

Proliferation of malassezia was measured as a zone of inhibition using a lawn spotting method.

All 3 supernatants were able to inhibit malassezia proliferation at pH below about 6.6.

Example 7:

Inhibition of malassezia furfur DSMZ 6170, malassezia restrictum (M.restrictum) CBS 7877 and She Zhuangma malassezia furfur (M.lobose) CBS 7874

Inhibition ：Zhang P.et al.(2015)Inter-strain interactions between bacteria isolated from vacuum-packaged refrigerated beef.Appl Environ Microbiol 81:2753–2761.doi:10.1128/AEM.03933-14 was evaluated using a sample application method adapted from the following lawn Arena,M.P.et al.(2016)Use of Lactobacillus plantarum Strains as a Bio-Control Strategy against Food-Borne Pathogenic Microorganisms.Frontiers in Microbiology 7(APR):1–10.doi:10.3389/fmicb.2016.00464.

The target strain malassezia furfur DSMZ 6170 was obtained from the Lebunz institute (Leibniz Institute) DSMZ. The target strains malassezia globosa (m.globosa) CBS 7874 and malassezia restrictively (m.restricta) CBS 7877 were obtained from the westerk fungus biodiversity institute (Westerdijk fungal biodiversity institute) CBS. The strain of interest was grown on modified Leeming Notmann (mLN) (ATCC medium No. 2737leeming & notman agar modified) agar and individual colonies were inoculated into 10mL of mLN broth. A lawn was created using the strain of interest culture and allowed to dry. The bacterial strain isolated from example 1 was stored in 10mL MRS broth. Subsequently, overnight LAB cultures were spotted onto plates and allowed to incubate. Pityrosporum ovale was grown at 30℃for about 10 days, malassezia restrictus was grown at 30℃for about 14 days, and Malassezia globosa was grown at 33℃for about 14 days. The zone of inhibition was measured in millimeters from the edge of the LAB colony to the onset of visible malassezia growth (clear zone). All plates were subjected to technical duplicate and repeated at different occasions.

LAB were screened against malassezia furfur DSMZ 6170, with 84 showing moderate inhibition and 18 showing high inhibition. 2 strains out of 18 were selected and subsequently tested for C.restrictus CBS 7877 and C.sphaeroides CBS7874 using lawn spotting. All 4 strains were able to inhibit the restriction malassezia CBS 7877 and malassezia globosa CBS7874.

TABLE 1

Spot-application assays were performed with malassezia. The inhibition/clearance zone is measured in millimeters as an average of the repetition between the bacterial colony periphery and the visible growth of malassezia as a target.

Example 8:

Trichophyton agar sample application assay

Antifungal activity against trichophyton was determined as described in example 7.

Trichophyton rubrum (Trichophyton rubrum) CBS189.69 is a nail isolate obtained from Werstdyke fungus biodiversity institute (Westerdijk Fungal Biodiversity Institute part) from the Royal academy of art and science (Royal NETHERLANDS ACADEMY of ARTS AND SCIENCE).

Trichophyton rubrum CBS 392.58 (novel Epidermophyton rubrum (Epidermophyton rubrum) Castell) is a skin (foot) isolate obtained from West dyke fungus biodiversity institute of the Royal art and science institute.

Trichophyton was grown on grignard maltose agar at 24 ℃ for 2 to 3 weeks.

TABLE 2

Spot-application assays were performed with trichophyton. Targeted, inhibition/clearance zone is measured in millimeters as an average of the repetition between the bacterial colony periphery and the visible growth of trichophyton

Example 9:

Inhibition by cell-free supernatant solution

Growth of malassezia (3 strains as described in example 7) with and without Cell Free Supernatant (CFS) from the LAB of example 2 was monitored using oCelloScope (BioSense Solutions ApS, denmark).

LAB cell cultures between 5 and 10mL were centrifuged (2,700 Xg, 10 min) and sterile filtered using a 0.2 μm syringe filter. Samples were taken from the LAB cultures after 24-72 hours.

A1:2 dilution was prepared from 10mL of Malassezia culture. 100 μl from each dilution was mixed with 100 μl pfs or dilutions thereof in a 96 well microtiter plate using 100, 75, 50, 25, 10 and 0% cfs. Volume was adjusted using MRS broth. As a positive control for inhibition, malassezia was treated with fluconazole (64. Mu.g/mL). All challenge assays were performed in triplicate and repeated on different occasions. The plates were incubated at 37℃in oCelloScope. The original image was taken every 2 half hours for up to 48 hours and the growth curve was plotted in excel and calculated as the average of three replicates.

After the experiment, the plates were visually inspected and the original images were visually inspected. The images were analyzed using UniExplorer PC software.

The results are shown in fig. 1-3, and demonstrate that cell-free supernatants obtained from three isolated strains (LB 555R, LB681R and LB 606R) have strong and significant inhibition of malassezia furfur DSM 6170 (fig. 1), malassezia restrictum CBS 7877 (fig. 2) and malassezia globosa CBS7874 (fig. 3) using a composition comprising: (i) Lactobacillus paracasei subspecies paracasei LB555R accession No. DSM 34249; (ii) lactobacillus plantarum, LB681R, accession No. DSM 34250; (iii) Pediococcus pentosaceus LB606R (DSM 33730), in comparison with compositions comprising the antifungal compound fluconazole (at a concentration of 64 μg/ml); and compared to untreated fungal solutions.

3 Hit strains were identified in the screen which inhibited fungal growth of at least 2 pathogenic fungi as lactic acid bacteria capable of growth, wherein the growth inhibition was significantly better than that observed for the known antifungal antibiotic fluconazole.

Example 10.

Analysis of short chain fatty acid production (SCFA)

Sample analysis was performed as follows. Cell-free supernatants from four strains (LB 555R; LB681R; LB606R; and LB 244R) grown in MRS medium for 24 hours at 37℃were acidified with hydrochloric acid, and deuterium labeled internal standard was added. Analysis was performed using a high polarity column (Zebron ^TM ZB-FFAP, GC cap. Column 30m x 0.25mm x 0.25 μm) mounted in GC (7890 b, agilent) coupled to a quadrupole detector (5977 b, agilent). The system was controlled by a chemical workstation (ChemStation) (Agilent). Raw data was converted to netCDF format using a chemical workstation (Agilent), after which the data was entered and processed in Matlab R2014b (Mathworks, inc.) using PARADISe software described by Johnsen et al (DOI: 10.1016/j.chroma.2017.04.052). The SCFA method is a GC-MS method specific for short chain fatty acids using highly polar columns and standards. Determination of short chain fatty acid (acetic acid) concentration in supernatant from each strain

	LB555R	LB681R	LB606R	LB244R
					Acetic acid	36.5mM	40.76mM	45.8mM	48.0mM

Example 11.

Different topical formulations for skin and nails

Powder and no-clean preparation

The components are as follows:

Rice (Oryza sative) (Rice) starch 49% (w/w)

Cassava starch 49% (w/w)

Lactobacillus paracasei subspecies paracasei LB555R 2% (w/w) (lyophilized powder, concentration 9x10 ⁹ CFU/g)

Powder and no-clean preparation

The components are as follows:

maize (Zea mays) (corn) starch 49% (w/w)

Cassava starch 49% (w/w)

Lactobacillus plantarum LB830R 2% (w/w) (lyophilized powder, concentration 8x10 ⁹ CFU/g)

External liquid ointment

The components are as follows:

Water 90% (w/w)

Fermentation lysate/metazoan (Lactobacillus plantarum, LB 681R) 10% (w/w)

External oil preparation:

The components are as follows:

jojoba oil 48% (w/w)

Sunflower seed oil 48% (w/w)

Lactobacillus plantarum LB681R 1% (w/w) (lyophilized powder, concentration 5X10 ⁹ CFU/g)

Lactobacillus paracasei subspecies paracasei LB555R 1% (w/w) (lyophilized powder, concentration 9x10 ⁹ CFU/g)

Inulin (FOS) 1% (w/w)

Tocopherol 1% (w/w)

Example 12.

Good diffusion measurement

LAB overnight cultures were prepared. LAB was grown in MRS broth and the fungi tested were grown for lawn spotting as described in the examples above. Wells were made in the plates and 50 μl of LAB culture was transferred into the wells. After incubation until fungal growth is seen, the zone of inhibition surrounding the well is checked.

Table 3: and (3) sample application and diffusion measurement of the lawn, wherein the inhibition zone is measured in mm.

Reference to the literature

US 4,654,207

US 5,019,376

US 3,929,678

US2,658,072

US2,438,091

US2,528,378

US 5,104,646

US 5,106,609

WO 2011/138450

FR 2908045

US 4,565,647

CTFA cosmetic ingredient Manual (Cosmetic Ingredient Handbook) (2004).

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PCT/RO/134 table

Claims

1. A composition comprising:

An acceptable carrier, which is a mixture of the components,

2. The composition of claim 1, wherein the composition comprises a nucleic acid and/or a nucleotide.

3. The composition according to any one of claims 1 or 2, wherein the composition does not comprise plant matter and/or fibrous matter.

4. A composition according to any one of claims 1-3, wherein the composition has the following pH: below pH 8.0, such as below pH 7.5, for example below pH 7.0, such as below pH 6.5, for example below pH 6.0, such as below pH 5.5, for example below pH 5.0, such as below pH 4.5, for example below pH 4.0, such as below pH 3.5, for example below pH 2.0, such as below pH 1.5, for example below pH 1.0.

5. The composition of any one of claims 1-4, wherein at least two active substances are produced by metabolism of an isolated live probiotic strain.

6. A composition according to any one of the preceding claims, wherein at least two active substances are produced by a single isolated probiotic strain.

7. A composition according to any of the preceding claims, wherein the active substance is selected from bacteriocins, organic acids, cell wall substances or combinations thereof.

8. Composition according to any one of the preceding claims, for use in the prevention and/or treatment of mycoses and/or dandruff in humans or animals.

9. The composition of claim 8, wherein the prevention and/or treatment of mycosis and/or dandruff in a human or animal is the prevention and/or treatment of a condition associated with proliferation of Malassezia (Malassezia) genus yeast on the scalp, hands, ears, feet, nails, or skin of a subject.

10. A topical hair care composition comprising a composition according to any one of claims 1 to 9 in combination with a shampoo base having a pH of less than 6.5, said shampoo base comprising at least one detersive surfactant selected from the group consisting of: anionic surfactant, nonionic surfactant, amphoteric surfactant, or combinations thereof.