Composition for treating, alleviating or preventing acne

Abstract

The present invention relates to a composition comprising one or more bacterial strains, wherein the one or more bacterial strains exhibit a stronger inhibitory effect on pathogenic Propionibacterium acnes strains than on non-pathogenic Propionibacterium acnes strains.

Description

Technical Field

[0001] The present invention relates to compositions comprising bacterial strains. In particular, the present invention relates to novel topical dermatological compositions comprising bacterial strains for treating; alleviating; inhibiting; preventing; and / or preventing the growth of pathogenic microorganisms. Background Art

[0002] Acne is one of the most common and well-established skin diseases, affecting approximately 90% of the world's population at some stage in their lifetime. The disease is a chronic inflammatory skin disease of the pilosebaceous glands of the skin with a multifactorial pathogenesis. It usually begins during adolescence due to hormonal changes and is further exacerbated by genetic factors.

[0003] Propionibacterium acnes (formerly known as Propionibacterium acnes, Propionibacterium acnes ) is a Gram-positive anaerobic bacterium that resides in sebaceous follicles, which are part of the natural flora of the pilosebaceous unit; however, elevated levels of specific ribotypes of P. acnes have been found in acne patients. In the microenvironment created in the context of acne, P. acnes thrives in the presence of increased sebum production, using it as a nutrient for growth. Increased levels of P. acnes within pores further promote the development of comedones.

[0004] P. acnes can stimulate inflammation by both metabolizing triglycerides and releasing free fatty acids. Inflammation is initiated by complement activation and the production of proinflammatory mediators that diffuse through the hair follicle wall. It is well known that P. acnes activates Toll-like receptor 2 (TLR2) on monocytes and neutrophils. Activation of TLR2 then leads to the production of multiple proinflammatory cytokines, particularly interleukin 12 (IL-12) and interleukin 8 (IL-8) and TNFα.

[0005] With increased pressure and recruitment of inflammatory mediators, microcomedones may rupture as immunogenic keratin, sebum, and bacteria are squeezed into the dermis. This stimulates a further exaggerated inflammatory response. The extent of inflammation and the respective cell types involved determine whether pustules, inflamed papules, nodules, or cysts will appear on the skin. In cases of acne with severe inflammation and tissue damage, hyperpigmentation and scarring may result.

[0006] There are various treatments for acne. Generally, acne treatments work by reducing oil production, speeding up skin cell turnover, fighting bacterial infections, reducing inflammation, or doing all four of these things. These types of acne treatments include over-the-counter topical treatments, antibiotics, oral contraceptives, and cosmetic surgery.

[0007] Acne lotions may dry up oil, kill bacteria, and promote shedding of dead skin cells. Over-the-counter (OTC) lotions are usually mild and contain benzoyl peroxide, sulfur, resorcinol, salicylic acid, or sulfur as their active ingredients. Antibiotics may cause side effects, such as upset stomach, dizziness, or skin discoloration. These drugs also increase the skin's sensitivity to sunlight and may reduce the effectiveness of oral contraceptives. For deep cysts, antibiotics may not be enough. Isotretinoin (Amnesteem, Claravis, Sotret) is a powerful medication that can be used to treat cystic acne or acne scars that don't respond to other treatments. However, isotretinoin has many side effects, such as dry skin, depression, severe stomach pain, and muscle / joint / back pain, and can cause birth defects in babies whose mothers use isotretinoin. Chemical peels and microdermabrasion may help control acne. These cosmetic procedures have traditionally been used to reduce the appearance of fine lines, sun damage, and minor facial scars and are most effective when combined with other acne treatments. They may cause temporary, severe redness, scaling and blistering, as well as long-term discoloration of the skin.

[0008] WO2017 / 220525 describes the beneficial effects of using a composition comprising probiotics, wherein a lactobacillus strain is selected to combat the development of common skin pathogens that cause acne, in particular Lactobacillus plantarum, Lactobacillus pentosus and / or Lactobacillus rhamnosus. It is described that the composition provided in WO2017 / 220525 comprises an acid, such as lactic acid produced by a selected lactobacillus strain as an important antimicrobial factor. A disadvantage of the present invention is that non-specific inhibition of Propionibacterium acnes (and natural skin microbiota) is provided, whereby two pathogenic ribosomal Propionibacterium acnes (leading to the development of common acne); and non-pathogenic ribosomal Propionibacterium acnes.

[0009] In addition to the negative side effects caused by currently available treatments, there are no treatments that target the modulation of the skin microbiota and target pathogenic ribotypes of P. acnes. Therefore, there is a need in the art for methods and treatments that specifically target pathogenic P. acnes, have limited antimicrobial effects on the natural skin microbiota, and thus support the natural skin microbiota in the fight against acne. Summary of the invention

[0010] Therefore, one object of the present invention relates to a composition comprising a bacterial strain which solves the above-mentioned problems of the prior art.

[0011] In particular, the present invention relates to a novel composition, preferably a novel topical dermatological composition, comprising a bacterial strain for treating; alleviating; inhibiting; preventing and / or preventing the growth of pathogenic microorganisms on the skin of mammals, such as humans, which solves the above-mentioned problems of the prior art, such as acne skin infections, acne skin diseases, skin inflammation, dry skin and scars caused by acne infections.

[0012] Thus, one aspect of the present invention relates to a composition comprising one or more bacterial strains, wherein the one or more bacterial strains exhibit a stronger inhibitory effect on pathogenic P. acnes strains than on non-pathogenic P. acnes strains.

[0013] Another aspect of the invention relates to a composition comprising one or more bacterial strains that are capable of co-aggregating with one or more pathogenic P. acnes strains relative to the ability of the one or more bacterial strains to co-aggregate with one or more non-pathogenic P. acnes strains.

[0014] Another aspect of the invention relates to a product comprising the composition according to the invention for use as a medicament.

[0015] Yet another embodiment of the present invention relates to a product comprising the composition according to the present invention for treating; alleviating, inhibiting; preventing and / or preventing the growth of pathogenic microorganisms.

[0016] Another embodiment of the present invention relates to a product comprising a composition according to the present invention comprising one or more bacterial species capable of co-aggregating one or more strains of Propionibacterium acnes for treating; alleviating, inhibiting; preventing and / or preventing acne vulgaris.

[0017] Another aspect of the present invention relates to a composition comprising one or more lactic acid bacteria species capable of coaggregating one or more Propionibacterium acnes strains, wherein the one or more lactic acid bacteria species may be selected from one or more bacterial strains having at least 95% genetic homology with one or more bacterial strains selected from the following group:

[0018] - LB356R (Lactobacillus plantarum ( Lactobacillus plantarum )LB356R, deposited as DSM33094)

[0019] - LB244R (Lactobacillus plantarum ( Lactobacillus plantarum )LB244R, deposited as DSM32996)

[0020] - LB349R (Leuconostoc mesenteroides ( Leuconostoc mesenteroides )LB349R, deposited as DSM33093)

[0021] - LB10G (Weissella viridis ( Weissella viridescens)LB10G, deposited as DSM32906).

[0022] Another aspect of the invention relates to the use of a composition comprising one or more bacterial strains for reducing or avoiding the attachment of one or more P. acnes strains to a surface and / or for reducing or avoiding the formation of a biofilm of one or more P. acnes on a surface. DETAILED DESCRIPTION

[0023] Before discussing the present invention in further detail, the following terms and definitions are first defined:

[0024] The term "ribotype" refers to a specific strain of P. acnes. Characteristics of ribotyped strains are described in Fitz-Gibbon et al. J. Investigative Dermatology 133:2152-60 (2013).

[0025] The term "phylotype" refers to a specific strain of P. acnes. Characteristics of phylotype strains are described in McDowell et al. PLoS ONE 8(9): e70897 (2013).

[0026] The term "pathogenic P. acnes" refers to ribotypes and phylotypes associated with acne disease and acne-infected skin.

[0027] A "therapeutically effective amount" of a compound of the subject methods of treatment means that amount of the compound in a formulation which, when administered as part of a dosage regimen required (to a mammal, preferably a human), alleviates symptoms, ameliorates symptoms, or slows the onset of a disease condition, according to clinically acceptable criteria for the disorder or condition being treated or for cosmetic purposes, e.g., at a reasonable benefit-risk ratio applicable to any medical treatment.

[0028] As used herein, the term "treating" or "treatment" includes reversing, alleviating or arresting the symptoms, clinical signs and underlying pathology of a disorder in a manner that improves or stabilizes the subject's disorder.

[0029] Therefore, P. acnes is a major commensal of human skin. It helps to keep the skin healthy by inhibiting the invasion of common pathogens such as Staphylococcus aureus and Streptococcus pyogenes. It does this by hydrolyzing triglycerides and releasing free fatty acids that contribute to the acidic pH of the skin surface. On the other hand, P. acnes has historically been associated with acne vulgaris, a chronic inflammatory disease of the pilosebaceous unit. A metagenomic study showed that P. acnes is the dominant bacterium in the pilosebaceous unit in healthy individuals and patients with acne.

[0030] It has been found that the population structure of P. acnes is different between the two groups of strains at the strain level. These findings suggest that human diseases associated with microorganisms are often caused by certain strains of a species rather than the entire species, consistent with studies of other diseases. P. acnes has been divided into three different types, first revealing two different P. acnes phenotypes, called type I and type II, which can be distinguished based on serum agglutination tests and cell wall sugar analysis. P. acnes types I and II were distinguished by monoclonal antibody typing. In addition, their phylogenetic analysis of P. acnes strains based on nucleotide sequences of the recA gene and the more variable hemolysin / cytotoxin gene (tly) showed that types I and II represent different lineages. Strains within the type I lineage can be further divided into two clades, called types IA and IB, both of which are associated with acne disease. Another phylogenetic group of P. acnes, called type III, is described later. Recent studies based on multilocus sequence typing (MLST) have further subdivided P. acnes into closely related clusters, some of which are associated with various diseases including acne. Results from the Human Microbiome Project (HMP) have generated knowledge about specific ribotypes of P. acnes strains associated with health and disease (McDowell et al. 2013, PLoS ONE 8(9):e70897, Dreno et al. 2018, JEADV 32(suppl. 2), 5-14. doi:10.1111 / jdv.15043). Specific ribotypes of P. acnes are known to be non-pathogenic and part of the natural healthy skin microbiota, whereas other specific ribotypes of P. acnes have been identified as pathogenic and dominant in acne infections. Thus, RT4 and RT5 have been shown to be strongly associated with acne, whereas RT6 is associated with healthy skin. Other P. acnes strains, including ribotypes RT7, RT8, RT9, and RT10, may also contribute to the development of the disease (Fitz-Gibbon et al. 2013, J. Invest. Dermatol. 133:2152-60. doi:10.1038 / jid.2013.21).

[0031] Acne-prone skin does not contain more P. acnes than healthy skin; it is an imbalance between pathogenic and non-pathogenic ribotypes of P. acnes.

[0032] Therefore, improved antimicrobial activity against pathogenic P. acnes would be advantageous.

[0033] The present invention relates to probiotics and compositions comprising probiotics for preventing or treating infections from pathogenic microorganisms, such as Proteobacterium acnes infections.

[0034] The inventors of the present invention surprisingly found that some bacterial strains can inhibit the growth of pathogenic ribotypes of P. acnes, while the growth of non-pathogenic ribotypes of P. acnes is significantly less affected, or not affected at all.

[0035] In one embodiment of the present invention, the one or more P. acnes strains may preferably be one or more pathogenic P. acnes strains.

[0036] One or more strains of Propionibacterium acnes, or one or more pathogenic Propionibacterium acnes strains, may be selected from Propionibacterium acnes ribotype RT4; ribotype RT5; ribotype RT7; ribotype RT8; ribotype RT9; and / or ribotype RT10. Preferably, one or more pathogenic Propionibacterium acnes strains may be selected from Propionibacterium acnes ribotype RT4; ribotype RT5; and / or ribotype RT8.

[0037] In one embodiment of the present invention, the one or more P. acnes strains may be selected from the group consisting of P. acnes HL110PA1, P. acnes HL053PA2, P. acnes HL043PA1 and / or P. acnes HL038PA1.

[0038] The present invention provides novel microbial strains and novel compositions that can inhibit the growth of pathogenic Propionibacterium acnes without causing skin irritation and dryness.

[0039] In the context of the present invention, the terms "inhibit" and "inhibition" may relate to complete or partial inhibition.

[0040] Therefore, a preferred embodiment of the present invention relates to a composition comprising one or more bacterial strains capable of co-aggregating one or more P. acnes strains.

[0041] Preferably, a preferred embodiment of the present invention relates to a composition comprising one or more bacterial strains, wherein the one or more bacterial strains exhibit a stronger inhibitory effect on pathogenic P. acnes strains than on non-pathogenic P. acnes strains.

[0042] The inventors surprisingly found that specific bacterial strains, in particular specific lactic acid bacterial strains, show a stronger effect on pathogenic Propionibacterium acnes strains than non-pathogenic Propionibacterium acnes strains. This discovery allows the provision of a composition comprising the specific bacterial strains suitable for treating, alleviating, inhibiting, preventing one or more pathogenic Propionibacterium acnes strains.

[0043] Preferably, the inhibition involves growth inhibition of Propionibacterium acnes strains, preferably inhibition of growth of pathogenic Propionibacterium acnes strains; co-aggregation inhibition of Propionibacterium acnes strains, preferably co-aggregation inhibition of pathogenic Propionibacterium acnes strains; and / or biofilm inhibition of Propionibacterium acnes strains, preferably biofilm inhibition of pathogenic Propionibacterium acnes strains. Most preferably, the inhibition involves inhibition of co-aggregation of Propionibacterium acnes strains, preferably inhibition of co-aggregation of pathogenic Propionibacterium acnes strains; and / or biofilm inhibition of Propionibacterium acnes strains, preferably biofilm inhibition of pathogenic Propionibacterium acnes strains.

[0044] In one embodiment of the present invention, the composition comprising one or more bacterial strains co-aggregates with one or more pathogenic P. acnes strains more strongly than the one or more bacterial strains co-aggregate with one or more non-pathogenic P. acnes strains.

[0045] The one or more bacterial strains may preferably be one or more lactic acid bacterial strains.

[0046] One or more bacterial strains (preferably one or more lactic acid strains) are capable of co-aggregating with one or more pathogenic P. acnes strains with strong aggregation strength. By visual inspection of the aggregation, the size of the aggregation can be determined as an indication of the aggregation strength of the one or more pathogenic P. acnes strains.

[0047] In a preferred embodiment of the present invention, one or more bacterial strains (preferably one or more lactic acid strains) are capable of co-aggregating with one or more non-pathogenic Propionibacterium acnes strains with a weak aggregation strength.

[0048] In a more preferred embodiment of the present invention, the composition and / or one or more bacterial strains (preferably one or more lactic acid bacteria) can co-aggregate with one or more pathogenic Propionibacterium acnes strains with a stronger aggregation intensity than with one or more non-pathogenic Propionibacterium acnes strains.

[0049] The formation of coaggregates can be scored visually from 1-5 (the higher the value, the stronger the aggregation intensity) using the following criteria:

[0050] 1: No aggregation

[0051] 2: Visualize initial aggregation

[0052] 3: Aggregate formation <0.5mm

[0053] 4: Aggregate formation > 0.5 mm and < 1 mm

[0054] 5: Aggregate formation > 1 mm

[0055] In an embodiment of the present invention, the inhibition of pathogenic Propionibacterium acnes strains is 10% stronger or more, such as 15% stronger or more, such as 20% stronger or more, such as 30% stronger or more, such as 40% stronger or more, such as 50% stronger or more, such as 75% stronger or more, such as 100% stronger or more, such as 150% stronger or more, such as 200% stronger or more, such as 300% stronger or more, relative to the inhibition of non-pathogenic Propionibacterium acnes strains.

[0056] In another embodiment of the invention, the inhibition of non-pathogenic P. acnes strains is 90% or less, such as 80% or less, such as 70% or less, such as 60% or less, such as 50% or less, such as 40% or less, such as 30% or less, such as 20% or less of the inhibition of pathogenic P. acnes strains.

[0057] In the context of the present invention, the terms "stronger" or "stronger intensity" are used interchangeably and, for example, in relation to synergistic aggregation, relate to an increased effect on non-pathogenic P. acnes strains relative to non-pathogenic P. acnes strains, wherein the stronger effect may result in the formation of larger aggregates comprising pathogenic P. acnes strains compared to the size of the aggregates formed comprising the non-pathogenic P. acnes strains. In relation to biofilm formation, a stronger effect may result in the formation of smaller biofilms with a reduced number of pathogenic P. acnes strains and / or the formation of biofilms with a higher proportion of non-pathogenic P. acnes strains relative to pathogenic P. acnes strains. In relation to an increased effect on growth, a stronger effect may result in an increased inhibition of the growth of pathogenic P. acnes strains relative to the growth of non-pathogenic P. acnes strains.

[0058] The inventors of the present invention surprisingly found that by improving the co-aggregation of the Propionibacterium acnes strains provided by the present invention, the formation of biofilms on surfaces can be limited, reduced or prevented. The formation of biofilms begins with the attachment of free-floating microorganisms to the surface. If these first colonists are not prevented from attaching to the surface, they can more permanently fix themselves on the surface and attract other microorganisms to attach, and the community may be growing and preparing for biofilm.

[0059] The compositions of the present invention unexpectedly exhibit the ability to limit, reduce or prevent:

[0060] 1) The first settlers of P. acnes strains that initially attach to surfaces; and / or

[0061] 2) Prevent P. acnes strains from attaching to colonizing microorganisms.

[0062] It is believed that one of the abilities of P. acnes to cause disease or infection may be related to attachment to human cell surfaces and the formation of P. acnes biofilms. This biofilm formation of P. acnes makes infection more difficult to prevent or treat because the biofilm formation protects the P. acnes bacteria from antimicrobial activity.

[0063] Biofilm formation by pathogenic P. acnes may provide intrinsic resistance to P. acnes strains, which is the innate ability of bacterial species to resist the activity of specific antimicrobial agents through their inherent structural or functional characteristics, which allows tolerance to specific drugs or antimicrobial compounds.

[0064] The inventors of the present invention surprisingly found that the bacterial strain according to the present invention, for example, the lactic acid strain of the present invention, is able to prevent the initial binding of Propionibacterium acnes to surfaces and inhibit the subsequent biofilm formation.

[0065] In one embodiment of the present invention, the bacterial strain according to the present invention, for example, the lactic acid bacterial strain according to the present invention, may be able to prevent the initial binding of Propionibacterium acnes to a surface and inhibit the subsequent production of a biofilm.

[0066] Preferably, the surface may be selected from a cell, a mammalian body, such as a human body, wherein the body may be the skin or a mucous membrane of a mammal, such as a human.

[0067] A preferred embodiment of the present invention relates to a composition comprising one or more bacterial strains capable of inhibiting, reducing, limiting and / or preventing attachment to the surface of one or more Propionibacterium acnes strains and / or inhibiting, reducing, limiting and / or preventing biofilm formation on the surface of one or more Propionibacterium acnes strains.

[0068] Preferably, the one or more strains of P. acnes that are inhibited, reduced, limited and / or prevented from attaching and / or forming a biofilm on a surface may be one or more pathogenic strains of P. acnes.

[0069] The attachment and / or biofilm formation of one or more P. acnes strains, preferably one or more pathogenic P. acnes strains, to a surface may be reduced by 15% (w / w) or more, such as by 25% (w / w) or more, such as by 50% (w / w) or more, such as by 75% (w / w) or more, such as by 90% (w / w) or more, such as by 95% (w / w) or more, such as by 98% (w / w) or more, relative to a sample without the addition of the bacterial strain.

[0070] In an embodiment of the present invention, one or more strains (preferably one or more lactic acid strains) can co-aggregate with a strong aggregation strength of 4 to 5 (on a scale of 1-5) after 1 hour and / or 24 hours. Preferably, one or more strains (preferably one or more lactic acid strains) can co-aggregate with a strong aggregation strength after 1 hour. The strong aggregation strength can be a score of 4 to 5 (on a scale of 1 to 5).

[0071] The faster a strong copolymer is formed, for example one or more pathogenic P. acnes strains may be, the better and more specific it is.

[0072] On the other hand, one or more bacterial strains, preferably one or more lactic acid bacterial strains, can co-aggregate with one or more non-pathogenic Propionibacterium acnes strains. Such co-aggregation with one or more non-pathogenic Propionibacterium acnes strains can be considered to have a weaker aggregation strength.

[0073] In one embodiment of the present invention, the one or more bacterial strains (preferably one or more lactic acid strains) are capable of co-aggregating with a weak aggregation strength, with a value of 1-3 (on a scale of 1-5) after 1 hour and / or 24 hours.

[0074] Due to the significant differences in the co-aggregation of one or more bacterial strains according to the present invention with pathogenic P. acnes strains and non-pathogenic P. acnes strains, wherein the strongest co-aggregation was found between one or more bacterial strains and pathogenic P. acnes strains, the inventors of the present invention surprisingly found that it is possible to treat; alleviate, inhibit; prevent; and / or prevent the development of mammalian acne vulgaris caused by pathogenic P. acnes strains. At the same time, the inventors surprisingly found that it is possible to simultaneously maintain non-pathogenic P. acnes strains (or portions thereof) as part of the healthy skin microbiota.

[0075] In one embodiment of the present invention, the one or more lactic acid bacteria strains may be a Lactobacillus strain; a Leuconostoc strain; and / or a Weissella strain. Preferably, the bacterial strain may be a Lactobacillus strain.

[0076] Preferably, the lactobacillus strain may be selected from Lactobacillus plantarum. In an embodiment of the present invention, the Lactobacillus plantarum may be selected from Lactobacillus plantarum LB356R, deposited as DSM33094, or from Lactobacillus plantarum LB244R, deposited as DSM32996.

[0077] Preferably, the Leuconostoc strain can be selected from Leuconostoc mesenteroides. In an embodiment of the present invention, the Leuconostoc mesenteroides can be selected from Leuconostoc mesenteroides LB349A deposited as DSM33093.

[0078] Preferably, the Vesella strain can be selected from Weissella viridis. In an embodiment of the present invention, Weissella viridis can be selected from Weissella viridis LB10G, deposited as DSM32906.

[0079] In another embodiment of the present invention, the one or more lactic acid bacteria strains capable of coaggregating one or more Propionibacterium acnes strains may be a combination of the following:

[0080] - Lactobacillus plantarum, preferably selected from Lactobacillus plantarum LB356R, deposited as DSM 33094 and / or selected from Lactobacillus plantarum LB244R, deposited as DSM 32996;

[0081] - Leuconostoc mesenteroides, preferably selected from Leuconostoc mesenteroides LB349A, deposited as DSM 33093; and / or

[0082] - Weissella viridis, preferably selected from Weissella viridis LB10G, deposited as DSM 32906.

[0083] Preferably, the one or more lactic acid bacteria species capable of coaggregating one or more Propionibacterium acnes strains may be selected from bacterial strains having at least 95% genetic homology with one or more bacterial strains in the group consisting of:

[0084] - LB356R (Lactobacillus plantarum LB356R, deposited as DSM33094)

[0085] - LB244R (Lactobacillus plantarum LB244R, deposited as DSM32996)

[0086] - LB349R (Leuconostoc mesenteroides LB349R, deposited as DSM33093)

[0087] - LB10G (Weissella viridis LB10G, deposited as DSM 32906).

[0088] A preferred embodiment of the present invention relates to a composition comprising one or more lactic acid bacteria species capable of coaggregating one or more Propionibacterium acnes strains, wherein the one or more lactic acid bacteria species may be selected from one or more bacterial strains having at least 95% genetic homology with one or more bacterial strains selected from the following group:

[0089] - LB356R (Lactobacillus plantarum LB356R, deposited as DSM33094)

[0090] - LB244R (Lactobacillus plantarum LB244R, deposited as DSM32996)

[0091] - LB349R (Leuconostoc mesenteroides LB349R, deposited as DSM 33093)

[0092] - LB10G (Weissella viridis LB10G, deposited as DSM 32906).

[0093] In another embodiment of the present invention, the one or more lactic acid bacteria species capable of co-aggregating one or more Propionibacterium acnes strains may be selected from bacterial strains having 96%; such as at least 97%; such as at least 98%; such as at least 99%; such as at least 99.5%; such as at least 99.8%; such as at least 99.9%; such as 100% (identical) genetic homology with a bacterial strain selected from the group consisting of:

[0094] - LB356R (Lactobacillus plantarum LB356R, deposited as DSM33094)

[0095] - LB244R (Lactobacillus plantarum LB244R, deposited as DSM32996)

[0096] - LB349R (Leuconostoc mesenteroides LB349R, deposited as DSM 33093)

[0097] - LB10G (Weissella viridis LB10G, deposited as DSM 32906).

[0098] In yet another embodiment of the present invention, the one or more lactic acid bacteria species capable of co-aggregating one or more Propionibacterium acnes strains may be selected from the group consisting of:

[0099] - LB356R (Lactobacillus plantarum LB356R, deposited as DSM33094)

[0100] - LB244R (Lactobacillus plantarum LB244R, deposited as DSM32996)

[0101] - LB349R (Leuconostoc mesenteroides LB349R, deposited as DSM 33093)

[0102] - LB10G (Weissella viridis LB10G, deposited as DSM 32906).

[0103] In the present context, the term "genetic homology" relates to the deviation of the genetic sequence of a bacterial strain relative to a deposited bacterial strain.

[0104] The effect of the bacterial strains (and / or compositions) according to the invention on pathogenic microorganisms may be significant.

[0105] The composition according to the invention is capable of inhibiting the growth of P. acnes. In one embodiment of the invention, the growth of pathogenic P. acnes may be reduced by at least 20%; for example by at least 30%, for example by at least 40%; for example by at least 50%, for example by at least 60%.

[0106] In a preferred embodiment, the bacterial strain according to the present invention may be an isolated bacterial strain.

[0107] - The present invention discloses microorganisms which are interconnected by functional relationships to form a unified concept according to the present invention, so that they share properties and / or effects, i.e. they inhibit the growth of pathogenic microorganisms such as Propionibacterium acnes, and / or reduce the colonization level of pathogenic microorganisms such as Propionibacterium acnes associated with skin diseases. These lactic acid bacteria include in particular selected from the group consisting of the following novel isolated microorganisms or their analogs, fragments, lysates, derivatives, mutants or combinations, which are deposited in the German Collection for Microorganisms and Cell Cultures:

[0108] - LB356R (Lactobacillus plantarum LB356R, deposited as DSM33094)

[0109] - LB244R (Lactobacillus plantarum LB244R, deposited as DSM32996)

[0110] - LB349R (Leuconostoc mesenteroides LB349R, deposited as DSM 33093)

[0111] - LB10G (Weissella viridis LB10G, deposited as DSM 32906).

[0112] The present invention includes compositions comprising at least one of these novel lactic acid bacteria and compositions comprising any combination of these strains and analogs, fragments, lysates, derivatives, mutants thereof.

[0113] Acne infections can be caused by Staphylococcus aureus ( Staphylococcus ) co-infections, such as Staphylococcus aureus, which can make the disease worse ( Staphylococcus aureus ).

[0114] In one embodiment of the invention, the composition according to the invention may also comprise an antimicrobial activity. In particular, the invention relates to a composition comprising one or more bacterial strains and having an antimicrobial activity against Staphylococci associated with acne co-infections.

[0115] In the context of the present invention, the term "bacterial strain" relates to a bacterial strain according to the present invention. Preferably, the bacterial strain relates to a lactic acid bacterial strain according to the present invention.

[0116] In the context of the present invention, the term "coaggregation" relates to the aggregation of pathogenic bacteria, such as the aggregation of pathogenic P. acnes strains with another bacterium, ie one or more bacterial strains according to the present invention, such as a lactic acid strain.

[0117] Preferably, the co-aggregates are clearly visible to the human eye.

[0118] In one embodiment of the present invention, the co-aggregates form aggregates with a particle size greater than 0.1 mm within 24 hours; for example, greater than 0.25 mm; for example, greater than 0.5 mm; for example, greater than 0.75 mm; for example, greater than 1.0 mm; for example, in the range of 0.1-0.5 mm; preferably, in the range of 0.5-1.0 mm; even more preferably greater than 1.0 mm.

[0119] In another embodiment of the present invention, the co-aggregates form aggregates with a particle size greater than 0.1 mm within 1 hour; for example, greater than 0.25 mm; for example, greater than 0.5 mm; for example, greater than 0.75 mm; for example, greater than 1.0 mm; for example, in the range of 0.1-0.5 mm; preferably, in the range of 0.5-1.0 mm; even more preferably greater than 1.0 mm.

[0120] Another aspect of the invention relates to a composition according to the invention for use in treating; alleviating, inhibiting; preventing and / or preventing the growth of pathogenic microorganisms.

[0121] The inventors of the present invention provide a therapeutic composition for treating or preventing infection, comprising a therapeutically effective concentration of one or more bacterial strains in a pharmaceutically acceptable carrier, suitable for administration to the gastrointestinal tract of a mammal and / or topical administration to the skin or mucosa of a mammal, wherein the one or more bacterial strains have the ability to inhibit pathogen growth, colonization rate and initial attachment of pathogens to the site of infection.

[0122] In an embodiment of the present invention, the composition may be a topical composition (for topical treatment) or an oral composition (for oral treatment) or a rectal composition (for rectal treatment). Preferably, the composition may be a topical composition, preferably a topical composition for topical treatment of acne vulgaris.

[0123] Acne treatment according to the present invention may be a combination therapy comprising topical treatment and oral treatment.

[0124] In another embodiment of the present invention, the composition may be a topical dermatological composition.

[0125] A preferred embodiment of the present invention relates to a composition comprising one or more bacterial strains according to the present invention.

[0126] The concentration of bacterial strains, preferably one or more live strains, can be 10 3 Up to 10 14 In the range of 10 colony forming units (CFU); for example, in the range of 10 5 -10 13 CFU range; for example, within 10 6 -10 12 CFU range; for example, 10 7 -10 11 CFU range; for example, 10 8 -10 10 CFU range; for example, about 10 9 CFU.

[0127] The bacterial strain (preferably one or more dead / inactivated strains, one or more strain lysates); the concentration of one or more strain metabolites, relative to the topical composition, can be in a concentration range of 0.001% (w / w) to 20% (w / w); for example, in the range of 0.01% (w / w) to 18% (w / w); for example, in the range of 0.1% (w / w) to 15% (w / w); for example, in the range of 0.5% (w / w) to 10% (w / w); for example, in the range of 1% (w / w) to 5% (w / w).

[0128] The bacterial strains defined herein may be provided in the composition according to the invention in the form of killed bacterial strains.The killed bacterial strains may be provided as whole dead cells or as a lysate, metabolite, derivative, analogue, fraction or extract obtained from the dead cells.

[0129] In one embodiment of the present invention, the one or more bacterial strains may be provided as one or more live strains, one or more dead or inactivated strains, one or more strain lysates; one or more strain metabolites or a combination thereof.

[0130] Preferably, the strain lysate may be provided by acid treatment, for example using HCl, and / or by sonication.

[0131] In the composition, the bacterial strain according to the present invention can be provided as one or more live strains, one or more dead strains or inactivated strains, one or more strain lysates; one or more strain metabolites; one or more analogs, one or more fragments, one or more derivatives, one or more mutants or a combination thereof (where the lysate is located); one or more strain metabolites; one or more analogs, one or more fragments, one or more derivatives, one or more mutants or a combination thereof (as obtained from the bacterial strain of the present invention) can treat; alleviate; inhibit; prevent; and / or prevent the growth of at least one pathogenic microorganism, for example, a Propionibacterium acnes strain.

[0132] P. acnes is a normal inhabitant of the human intestinal microbiota and colonizes the intestine. Therefore, intestinal colonization by pathogenic ribotypes may also affect the skin microbiota, and thus probiotics with anti-P. acnes activity may be advantageously used as oral compositions.

[0133] In one embodiment of the present invention, wherein the composition may be a topical composition and / or an oral composition, preferably the composition is a topical composition.

[0134] In one embodiment of the present invention, wherein the composition may be a topical composition and / or an oral composition, preferably the composition is an oral composition.

[0135] In embodiments of the invention, the composition may have an acidic pH value, preferably the pH value of the composition is pH 6.5 or lower; for example, pH 6 or below; for example, pH 5.5 or below; for example, pH 5 or below; for example, pH 4.5 or below; for example, in the range of pH 4-6.5; for example, in the range of pH 4.5-6.0; for example, in the range of pH 4.7-5.5.

[0136] In another embodiment of the invention, the composition, when applied to a surface such as human skin, may result in an acidic pH value on the surface (skin), preferably a pH value of pH 6.5 or below; such as pH 6 or below; such as pH 5.5 or below; such as pH 5 or below; such as pH 4.5 or below; such as in the range of pH 4-6.5; such as in the range of pH 4.5-6.0; such as in the range of pH 4.7-5.5.

[0137] The composition according to the present invention may preferably comprise a pharmaceutically or cosmetically acceptable carrier or excipient.In an embodiment of the present invention, the composition may be provided in a solid form, a liquid form, a viscous form, an emulsion or as a dried form.

[0138] The oral composition may preferably be formulated as a paste, a soft gelatin capsule, a hard gelatin capsule, a powder, talc, granules, beads, lozenges, effervescent tablets, pastilles, buccal tablets, chewable tablets, sublingual tablets, oils, liquids, solutions, tinctures, emulsions, juices, concentrates, syrups, sprays, mists, drinking ampoules, gels, chewing gums, tablets, coated pills or as a food or feed product or a beverage.

[0139] Compositions for topical application may preferably be formulated as a paste; talc; a powder; an emulsion; a custard; a foam; a cream; an oil, a serum or an ointment.

[0140] In one embodiment of the present invention, the topical composition may be a powder composition comprising hydrated magnesium silicate (talc) and at least one bacterial strain of the present invention.

[0141] In a further embodiment according to the invention, a powder composition for topical application comprises hydrated magnesium silicate, at least one carbohydrate and at least one bacterial strain according to the invention.

[0142] In preferred embodiments, the topical composition may be formulated as a lotion; a custard; a foam; a cream; an ointment, an oil, a serum, a gel or an emulsion.

[0143] In a preferred embodiment, the topical composition is a vegetable oil.

[0144] In a preferred embodiment, the treatment is a combination treatment of a topical composition and an oral composition comprising the bacterial strain of the invention.

[0145] In addition to the bacterial strain according to the present invention, the composition may further comprise other probiotics, prebiotics, antimicrobials, antibiotics or other active antimicrobial substances and / or preferably may also contain one or more substances selected from the following: antioxidants, vitamins, coenzymes, fatty acids, amino acids and cofactors.

[0146] In another embodiment of the present invention, the bacterial strain according to the present invention can be combined with:

[0147] - Therapeutically effective doses of antibiotics. As combination therapy or after antibiotic therapy;

[0148] - Therapeutic concentrations of antibiotics, including but not limited to: fusidic acid; vancomycin; gentamicin; oxacillin; tetracycline; nitrofurantoin; chloramphenicol; clindamycin; trimethoprim-sulfamethoxazole; members of the cephalosporin antibiotic family (e.g., cefaclor, cefdroxil, cefixime, cefprozil, ceftriaxone, cefuroxime, cephalexin, loracarbef, etc.); members of the penicillin antibiotic family (e.g., ampicillin, amoxicillin / clavulanate, bacancillin, cloxacillin, penicillin VK, etc.); members of the fluoroquinolone antibiotic class (e.g., ciprofloxacin, grepafloxacin, levofloxacin, lomefloxacin, norfloxacin, ofloxacin, sparfloxacin, trovafloxacin, etc.); or members of the macrolide antibiotic family (e.g., azithromycin, erythromycin, etc.);

[0149] - a therapeutically effective dose of an anti-inflammatory drug. As a combination therapy or after treatment; and / or

[0150] - Therapeutic concentrations of anti-inflammatory drugs.

[0151] In an embodiment of the present invention, the composition may be a medicine, a veterinary or a food or a food supplement or a food supplement composition. The composition (preferably for oral administration) may preferably comprise one or more thickeners, and / or one or more sweeteners and / or one or more artificial sweeteners, wherein the thickener is preferably selected from cellulose ethers, polysaccharides, selected from the group comprising: xanthan gum, gelatin, highly dispersed silicon dioxide, starch, carrageenan, alginate, tragacanth gum, agar, gum arabic, pectin and polyvinyl esters, and the sweetener is selected from the group comprising: glucose, fructose, sucrose, glucose syrup, sorbitol, mannitol, xylitol, maltitol, steviol glycosides, saccharin, sodium cyclamate, acesulfame K and / or aspartame.

[0152] Preferred foods and nutritional supplements in the sense of the present invention may include effervescent tablets, vitamin tablets, dietary supplements, mineral tablets, trace element tablets, beverage powders, beverages, juices, milk drinks, yogurt, mineral water, non-carbonated water, candies, chewable tablets, juices or syrups, coated pills and lozenges and aerosols.

[0153] In addition, the composition may further comprise builders, enzymes, electrolytes, pH adjusters, thickeners, prebiotics, fluorescent whitening agents, graying inhibitors, foam regulators and / or colorants.

[0154] There is no disclosure in the prior art of using probiotic strains to prevent or treat infection with specific pathogenic acne infections caused by pathogenic ribotypes of Propionibacterium acnes.

[0155] It was completely surprising that a group of lactic acid bacteria could be identified that had the same advantageous properties. No bacteria, specifically no lactic acid bacteria, combined with the properties of treating; alleviating, inhibiting; preventing; and / or preventing the growth of pathogenic microorganisms, while also being non-pathogenic and not causing any damage or impact on the skin or microbiota.

[0156] It should be understood that, hereinafter, preferred embodiments related to one broad aspect of the invention are equally applicable to each of the other broad aspects of the invention described above. It will be further understood that the preferred embodiments described below may be combined unless the context indicates otherwise. As used herein, the term topical includes preparations that are applied to body surfaces (e.g., skin or mucous membranes).

[0157] Embodiments of the invention relate to compositions according to the invention for treating; alleviating, inhibiting; preventing and / or preventing the growth of pathogenic microorganisms.

[0158] One embodiment of the invention relates to the use of a composition according to the invention for healing scars and wounds caused by bacterial infections.

[0159] More preferably, the present invention may provide a composition as defined herein for use in the treatment, alleviation, inhibition or prevention of one or more pathogenic bacterial infections in a mammal.

[0160] More preferably, the present invention may provide a composition as defined herein for use in preventing the growth of pathogenic microorganisms.

[0161] The bacterial infection may preferably be a Propionibacterium acnes infection of a mammal.

[0162] One embodiment of the present invention relates to a composition as defined herein for use in treating; alleviating; inhibiting; and / or preventing a disease caused by infection with Propionibacterium acnes.

[0163] In another embodiment of the present invention, the composition comprises at least one bacterial strain (or a lysate, metabolite, derivative, analog, fraction or extract thereof) for treating skin infections caused by Propionibacterium acnes.

[0164] A preferred embodiment of the invention relates to compositions for use as a prophylactic or medical treatment of P. acnes infection.

[0165] Microorganisms can advantageously be present in the composition in a live or killed / dead form. Bacterial strains can be provided in encapsulated, microencapsulated, spray-dried and / or lyophilized form. In addition, bacterial strains can be provided in the form of cell lysates, metabolites, derivatives, analogs, fractions or extracts.

[0166] The microorganisms may advantageously be present in the composition in live form.The bacterial strains may be provided in an encapsulated, microencapsulated, spray-dried and / or freeze-dried form in an oil formulation.

[0167] In a preferred embodiment of the present invention, the oil is selected from almond oil, hemp oil, CBD oil, cannabis oil, evening primrose oil, borage oil, almond sweet oil, rosehip oil, jojoba oil, jojoba oil, chamomile oil, calendula oil, sea buckthorn oil, safflower oil and sesame oil.

[0168] In a preferred embodiment of the present invention, the composition comprises at least one selected from almond oil, hemp oil, CBD oil, hemp oil, evening primrose, borage oil, almond sweet oil, rosehip oil, jojoba gold oil, jojoba oil, chamomile oil, calendula oil, sea buckthorn oil, safflower oil and / or sesame oil.

[0169] In a preferred embodiment of the invention, the composition comprises at least 10% (w / w) jojoba oil or jojoba wax or jojoba gold oil.

[0170] In embodiments of the present invention, the bacterial strain may be present in the composition in an amount of 0.001% (w / w) to 20% (w / w) by weight; for example in the range of 0.01% (w / w) to 18% (w / w); for example in the range of 0.1% (w / w) to 15% (w / w); for example in the range of 0.5% (w / w) to 10% (w / w); for example in the range of 1% (w / w) to 5% (w / w), relative to the topical composition.

[0171] A preferred embodiment of the present invention involves administering approximately 1×10 3 Up to 1×10 14 CFU of viable bacteria, more preferably about 1 x 10 4 Up to 1×10 10 , most preferably about 5×10 4 Up to 1×10 9 CFU of live bacteria. When the condition to be treated involves an antibiotic-resistant pathogen and the patient is an adult, a typical dose is approximately 1 × 10 2 Up to 1×10 14 CFU of live bacteria, preferably about 1×10 8 Up to 1×10 10 , more preferably about 2.5×10 8 Up to 1×10 10 CFU viable bacteria per day.

[0172] The present invention relates to novel bacterial strains and the general references in the claims are to living cells, dead / killed cells and lysates, metabolites, derivatives, analogs, fractions or extracts and compositions comprising these living cells, dead / killed cells and lysates, metabolites, derivatives, analogs, fractions or extracts.

[0173] The compositions of the present invention can be used to treat; alleviate, inhibit; prevent; and / or prevent the growth of pathogenic microorganisms, such as Propionibacterium acnes, which exist in children, adolescents, healthy people, women, the elderly, immunosuppressed people, single or recurrent Propionibacterium acnes infections and / or people infected with antibiotic-resistant bacteria.

[0174] Therefore, the composition of the present invention can be used to prepare a medicament useful for treating or preventing the growth of Propionibacterium acnes. In an embodiment of the present invention, the composition can be used for treatment or prevention, for example in combination with a probiotic and / or prebiotic composition.

[0175] The combination of the composition according to the invention and the probiotic strain provides a combined composition capable of inhibiting the growth of P. acnes in a co-culture by reducing the growth of P. acnes by at least 50% compared to growth in the absence of the bacterial strain, wherein the growth is measured as colony forming units of P. acnes.

[0176] In embodiments of the invention, the bacterial strain as defined herein may be the only bacteria present in the composition. A composition comprising only the bacterial strain as defined herein shows at least a 50% reduction in the growth of P. acnes compared to growth without the bacterial strain.

[0177] In another embodiment of the invention, one or more bacterial strains as defined herein constitute at least 10% (w / w); such as at least 25% (w / w); such as at least 50% (w / w); such as at least 75% (w / w); such as at least 85% (w / w); such as at least 90% (w / w); such as at least 95% (w / w); such as at least 98% (w / w); such as at least 100% (w / w) of the total bacterial content of the composition.

[0178] A "reduction" in growth can be "statistically significant" and can include a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% reduction compared to a growth period in the absence of the bacterial strain of the invention.

[0179] In one embodiment of the invention, growth inhibition can be determined as a reduction in growth of at least 25%. Preferably, growth inhibition is determined as a reduction in growth of at least 50%. Even more preferably, growth inhibition is determined as a reduction in growth of at least 90%.

[0180] A "reduction" in the number of microorganisms can be "statistically significant" compared to the number of CFU / ml in the absence of the bacterial strain of the invention and can include a 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99%, 99.9% or 100% reduction.

[0181] The number of microorganisms was measured as colony forming units (CFU / ml).

[0182] The microorganisms according to the invention may preferably be in isolated or purified form, wherein the term "isolated" particularly means that the lactic acid bacteria are derived from their culture medium, including for example their natural culture medium. The term "purified" is not limited to absolute purity.

[0183] In an embodiment of the invention, the probiotic strains may be used as live isolated microorganisms in a stable form. Suitable methods for stabilization are known to those skilled in the art and include freeze drying or lyophilization involving different cryoprotectants.

[0184] In another embodiment of the present invention, the strain can be used as a live isolated strain.

[0185] Preferably, the strain can be used as a live isolated stable strain. Even more preferably, the strain can be used as a live isolated strain stabilized by lyophilization. Even more preferably, the strain can be used as a live isolated strain stabilized by lyophilization and comprising a cryoprotectant.

[0186] The present invention relates to living and / or dead (killed) bacterial strains, both forms are included within the scope of the present invention.

[0187] Suitable killing methods (e.g., biological, chemical or physical killing methods) are well known to those skilled in the art. However, in the present case, the bacterial strains can also be used in lyophilized form. The killed form of the microorganisms can include the fermentation broth and any metabolites present.

[0188] The term "killed" or "dead" relates to inactivated lactic acid bacteria that are unable to undergo cell division and do not have any metabolic activity. Dead or killed lactic acid bacteria may have an intact or disrupted cell membrane.

[0189] "Lysates", "derivatives", "analogs", "fractions" or "extracts" can be obtained from dead or killed lactic acid bacteria. These lysates, fractions, derivatives, analogs and extracts preferably have the property of reducing the transfer of pathogenic microorganisms between the surface of a first subject and the surface of a second subject, wherein "lysates" and the term "extracts" particularly refer to solutions or suspensions of microbial cells according to the present invention in an aqueous medium and include, for example, macromolecules such as DNA, RNA, proteins, peptides, lipids, carbohydrates, etc. and cell debris. The lysate preferably includes cell walls or cell wall components, including binding receptors. Methods for producing lysates are well known to those skilled in the art and include, for example, the use of "French press" or enzymatic lysis, ball mills with glass beads or iron beads. Cells can be broken by enzymatic, physical or chemical methods. Examples of enzymatic cell lysis may include individual enzymes as well as enzyme mixtures, such as proteases, proteinase K, lipases, glycosidases; chemical lysis may be induced by ionophores, detergents (such as SDS), acids or bases; physical methods can also be implemented by using high pressure, such as French presses, osmotic pressure, temperature or alternating hot and cold. Furthermore, chemical, physical and enzymatic methods can of course be combined.

[0190] In a preferred embodiment, the composition and / or bacterial strain according to the present invention is suitable for treating; alleviating, inhibiting; preventing diseases associated with infection with pathogenic microorganisms in mammals.

[0191] The present invention also relates to a method of reducing the number of Propionibacterium acnes on the skin of a patient infected with acne.

[0192] In an embodiment of the present invention, a composition comprising at least one bacterial strain can be used to control the number of Propionibacterium acnes on the skin of a patient with an inflammatory skin disease to a level at which Propionibacterium acnes does not cause skin infection.

[0193] In yet another embodiment of the present invention, a composition comprising at least one bacterial strain is useful for controlling the number of Propionibacterium acnes on the skin of a patient suffering from an inflammatory skin disease, wherein the level of Propionibacterium acnes is less than about 10 7 CFU / cm 3 skin.

[0194] It will be clear to a person skilled in the art that all statements of ranges given here and in the present invention, characterized by terms such as "about" or "approximately", do not necessarily indicate an exact numerical range, but even slight deviations up or down about the indicated numbers are still within the scope of the present invention. In embodiments of the present invention, minor deviations may include deviations of 5% or less, such as 4% or less, such as 4% or less, such as 3% or less, such as 2% or less, such as 1% or less.

[0195] In embodiments of the invention, biologically pure cultures of one or more bacterial strains of the invention may be provided.

[0196] The term "effective amount" may depend on the context in which it is used. In the case of administering a composition to reduce the risk of a P. acnes infection and / or administering a composition to reduce the severity of a P. acnes infection in a subject and / or reduce the amount of P. acnes, an effective amount of the composition described herein is an amount sufficient to treat and / or ameliorate a P. acnes infection and to reduce the severity of a P. acnes infection and / or reduce the likelihood of such infection. The reduction in the amount of pathogenic P. acnes in a subject may be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 99.9% reduction in the severity of a P. acnes infection or the likelihood of being infected.

[0197] An effective amount may be administered as a composition in one or more administrations.

[0198] An effective amount of the composition can be administered topically, orally, or a combination thereof. Topical administration is preferred.

[0199] In one embodiment of the invention, the composition comprises at least one bacterial strain according to the invention and a prebiotic.

[0200] "Prebiotics" are non-digestible food ingredients that promote the growth of specific microorganisms. "Synbiotics" are compositions that contain at least one probiotic and at least one prebiotic. Such compositions are understood to promote the growth of beneficial bacteria, such as probiotics. Thus, powerful synbiotics are based on the combination of specific strains of probiotics with carefully selected prebiotics. They can bring important health benefits to mammals.

[0201] According to another aspect of the present invention, there is provided a probiotic composition comprising probiotic microorganisms and at least one further active ingredient.

[0202] Prebiotics are chemical products that induce the growth and / or activity of commensal microorganisms (e.g., bacteria and fungi) that contribute to the health of the host. Prebiotics are non-digestible carbohydrates that pass through the upper gastrointestinal tract undigested and stimulate the growth and / or activity of beneficial bacteria that colonize the large intestine or the skin microbiome.

[0203] Some oligosaccharides used as prebiotics are fructooligosaccharides (FOS), xylooligosaccharides (XOS), polydextrose, pectin, galacto-oligosaccharides (GOS) or human milk oligosaccharides (HMO). In addition, disaccharides such as lactulose or some monosaccharides such as lactose or tagatose can also be used as prebiotics.

[0204] In an embodiment of the present invention, at least one prebiotic compound may be included in the composition of the present invention. In a very broad concept, prebiotics are all compounds that can be metabolized by probiotics.

[0205] Preferably, prebiotics are indigestible or difficult to digest by mammals. It is believed that prebiotics, many of which are indigestible carbohydrates, promote the growth of probiotics. Prebiotics are naturally present in, for example, cabbage, onions, whole grains, bananas, garlic, honey, leeks, artichokes, fortified foods and beverages, and dietary supplements. Prebiotics are well known in the art, and when used in the present invention, there is no particular limitation on the prebiotics themselves.

[0206] However, in an embodiment, at least one prebiotic product in the composition is selected from the following compounds and compositions: non-digestible carbohydrates, β-glucan, manno-oligosaccharide, inulin, oligofructose, human milk oligosaccharide (HMO), galacto-oligosaccharide (GOS), lactulose, lactofructo-oligosaccharide, galacto-triose, oligofructose (FOS), cellobiose, cellodextrin, cyclodextrin, maltitol, lactitol, glycosilsucrose, betaine, vitamin E or variants thereof (wherein the variant is selected from α, β, γ, δ tocopherol, tocotrienol and tocopherol monoenol). Optionally, manno-oligosaccharide and / or inulin may be preferred. HMOs may include lacto-N-tetraose, lacto-N-fucopentaose, lacto-N-triose, 3'-sialyllactose, lacto-N-neofucopentaose, sialic acid, L-fucose, 2-fucosyllactose, 6'-sialyllactose, lacto-N-neotetraose, and 3-fucosyllactose.

[0207] Prebiotics may also be used in the topical compositions of the present invention.

[0208] In one embodiment, at least one of the following prebiotic compounds is used in the topical composition of the present invention: lactose, β-glucan, manno-oligosaccharide, inulin, fructo-oligosaccharide, galacto-oligosaccharide (GOS), lactulose, lactofructo-oligosaccharide, galacto-triose, fructo-oligosaccharide (FOS), cellobiose, cellodextrin, cyclodextrin, maltitol, lactitol, glucosylsucrose, betaine, vitamin E or variants thereof (wherein the variant is selected from α, β, γ, δ tocopherol, tocotrienol and toco-monoenol), lacto-N-tetraose, lacto-N-fucosylpentaose, lacto-N-triose, 3'-sialyllactose, lacto-N-neofucylpentaose, sialic acid, 2-fucosyllactose, 6'-sialyllactose, lacto-N-neotetraose and 3-fucosyllactose. Optionally, lactose and / or manno-oligosaccharide and / or inulin may be preferred.

[0209] D- and L-fucose strengthen the natural defenses of the skin, stimulate the epidermal immune defenses and / or prevent and / or treat autoimmune diseases of the skin. In an embodiment of the invention, the composition comprises D- or L-fucose.

[0210] In one embodiment of the present invention, the composition further comprises L-fucose at a concentration of 10 mM to 500 mM in the composition.

[0211] According to still further features in the described preferred embodiments, the composition further comprises at least one active ingredient.

[0212] In one embodiment of the present invention, the composition comprises at least one bacterial strain of the present invention in combination with at least one other probiotic microorganism selected from another bacterium, a yeast or a mold.

[0213] The composition according to the invention may comprise at least one bacterial strain according to the invention in combination with at least one additional probiotic microorganism, wherein the at least one additional probiotic microorganism may be selected from, but not limited to: Bifidobacterium lactis DSM10140, Bifidobacterium lactis LKM512, Bifidobacterium lactis DSM20451, Bifidobacterium bifidum BB-225, Bifidobacterium adolescentis BB-102, Bifidobacterium breve BB-308, Bifidobacterium longum BB-536 from Zaidanhojin Nihon Bifizusukin Senta (Japan Bifidobacterium Center), Bifidobacterium NCIMB 41675 described in EP2823822.Bifidobacterium bifidum BB-225, Bifidobacterium adolescentis BB-102, Bifidobacterium breve BB-308, Bifidobacterium lactis HN019 (Howaru) available from DuPont Nutrition Biosciences ApS, Bifidobacterium lactis DN 173 010 available from Groupe Danone, Bifidobacterium lactis Bb-12 available from Chr. Hansen A / S, Bifidobacterium lactis 420 available from DuPont Nutrition Biosciences ApS, Bifidobacterium breve Bb-03, Bifidobacterium lactis BI-04, Bifidobacterium lactis Bi-07 available from DuPont Nutrition Biosciences ApS, Bifidobacterium bifidum Bb-02, Bifidobacterium bifidum Bb-06, Bifidobacterium longum KC-1, and Bifidobacterium longum 913 (DuPont Nutrition Biosciences ApS), Bifidobacterium breve M-16V (Morinaga) and / or lactobacilli with probiotic action, and may be any of the following strains; Lactobacillus rhamnosus LGG (Chr. Hansen), Lactobacillus acidophilus NCFM (DuPont Nutrition Biosciences ApS), Lactobacillus bulgaricus 1260 (DuPont Nutrition Biosciences ApS), Lactobacillus paracasei Lpc-37 (DuPont NutritionBiosciences ApS), Lactobacillus rhamnosus HN001 (Howaru) available from DuPont Nutrition Biosciences ApS, Streptococcus thermophilus 715 and Streptococcus thermophilus ST21 available from DuPont Nutrition Biosciences ApS, Lactobacillus paracasei subsp., paracasei CRL431 (ATCC 55544), Lactobacillus paracasei strain F-19 from Medipharm, Inc., Lactobacillus paracasei LAFTI L26 (DSM Food Specialties, The Netherlands) and Lactobacillus paracasei CRL 431 (Chr. Hansen), Lactobacillus acidophilus PTA-4797, Lactobacillus salivarius Ls-33 and Lactobacillus curvatum 853 (DuPontNutrition Biosciences ApS).Lactobacillus paracasei subsp. rhamnosus LC705, Lactobacillus DSM15527 (Bifodan), Lactobacillus DSM15526 (Bifodan) described in Finnish Patent 92498, Valio Oy, Lactobacillus rhamnosus GG (LGG) (ATCC 53103) and Lactobacillus rhamnosus LC705 (DSM7061) described in U.S. Patent 5,032,399, Propionibacterium freudenreichii subsp. schizophylla PJS (DSM 7067) described in more detail in Finnish Patent 92498, Valio Oy, Nitrosomonas D23 (ABIome), Staphylococcus hominis A9, C2, AMT2, AMT3, AMT4-C2, AMT4-Gl and / or AMT4-D12 (all from Matrisys Bioscience), Staphylococcus epidermidis (. Staphylococcus epidermidis ) M034, M038, All, AMT1, AMT5-C5 and / or AMT5-G6 (all from Matrisys Bioscience), Lactobacillus plantarum YUN-V2.0 (BCCM LMG P-29456), Lactobacillus pentosus YUN-V1.0 (BCCN LMG P-29455), Lactobacillus rhamnosus YUN-S1.0 (BCCM LMG P-2961), Weissella viridis LB10G (DSM 32906), Lactobacillus paracasei ( Lactobacillus paracasei )LB113R(DSM 32907)、Lactobacillus plantarum LB244R (DSM 32996)、Lactobacillus paracasei LB116R(DSM 32908)、Enterococcus faecium( Enterococcus faecium ) LB276R (DSM 32997), Lactobacillus plantarum LB316R (DSM 33091), Lactobacillus plantarum LB356R (DSM 33094), Lactobacillus plantarum LB312R (DSM 33098) and / or any combination thereof.

[0214] In an embodiment of the invention, the composition comprises at least one bacterial strain as defined herein in combination with at least one strain selected from lactic acid bacteria capable of improving the integrity of tight junctions.

[0215] In another embodiment of the invention, the composition comprises at least one bacterial strain as defined herein in combination with at least one strain selected from the group consisting of Lactobacillus rhamnosus LGG (Chr. Hansen), Lactobacillus acidophilus NCFM (DuPont), Lactobacillus salivarius Ls-33 (DuPont), Propionibacterium jannaschii P63 (DuPont), Bifidobacterium lactis 420 (DuPont) and Lactobacillus acidophilus La-14 (DuPont); and / or cell lysates and / or soluble metabolites of probiotic strains.

[0216] As used herein, and as is well known in the art, "treatment" is an approach for obtaining beneficial or desired results, including clinical results. For the purposes of this subject matter, beneficial or desired clinical results include, but are not limited to, alleviation or amelioration of one or more symptoms, reduction in the extent of the disease, a stable (i.e., non-worsening) state of the disease, prevention of the disease, delay or slowing of disease progression, and / or improvement or palliation of the disease state.

[0217] A reduction can be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98% or 99% reduction in the severity of a complication or symptom.

[0218] In one embodiment of the present invention, a method for treating mammalian skin may be provided. The method comprises administering a therapeutically effective amount of at least one bacterial strain to a subject (e.g., a mammal) in need thereof, thereby treating the skin to reduce colonization and / or carrier levels and / or infection.

[0219] In another embodiment of the invention, skin colonization may be caused by antibiotic resistant microorganisms.Skin colonization may be caused by resistant Propionibacterium acnes.

[0220] The present invention may provide several advantages. In particular, antimicrobial treatments that do not use conventional antimicrobial agents are desirable due to the potential for the development of resistant microbial species that may adversely affect the use of anti-acne products. Thus, the present invention does not contribute to the development of future generations of antibiotic-resistant pathogens.

[0221] The present invention may provide several advantages. In particular, since the use of anti-acne products can result in significant drying of the skin, which can be a disadvantage when using anti-acne products, there is a need for an antimicrobial treatment that does not cause any skin drying. Thus, the present invention does not cause skin drying.

[0222] The present invention may provide several advantages. In particular, since the use of anti-acne products may be adversely affected by the potential for skin irritation and / or inflammation, it would be desirable to have an antimicrobial treatment that does not cause skin irritation or further inflammation.

[0223] The present invention may provide several advantages. In particular, there is a need for antimicrobial treatments that do not cause any discoloration or sun sensitivity to the skin, as the resulting skin discoloration (pigmentation) and sun sensitivity have adverse effects on the use of anti-acne products. Thus, the present invention does not cause coloration or sun sensitivity. Another advantage of the present invention is that the present invention reduces the appearance of pigmentation and discoloration, as a further benefit of antimicrobial activity against Propionibacterium acnes.

[0224] A preferred embodiment of the present invention relates to a pharmaceutical composition comprising the composition according to the invention for use as a medicament.

[0225] Furthermore, a preferred embodiment of the present invention relates to a pharmaceutical composition comprising the composition according to the present invention for use in treating; alleviating, inhibiting; preventing; and / or preventing the growth of pathogenic microorganisms.

[0226] Furthermore, a preferred embodiment of the present invention relates to a pharmaceutical composition comprising a composition according to the present invention comprising one or more bacterial species capable of co-aggregating one or more strains of Propionibacterium acnes for treating; alleviating, inhibiting; preventing; and / or preventing acne vulgaris.

[0227] Yet another embodiment of the present invention relates to a method of treating; alleviating, inhibiting; preventing; and / or preventing the growth of pathogenic microorganisms, in particular one or more pathogenic Propionibacterium acnes strains, e.g., acne vulgaris in a mammal such as a human, the method comprising administering to a mammal in need thereof a therapeutically effective amount of a composition as defined herein comprising one or more bacterial species, e.g., one or more lactic acid bacteria as defined herein, e.g., one or more bacterial strains selected from the group consisting of:

[0228] - LB356R (Lactobacillus plantarum LB356R, deposited as DSM33094)

[0229] - LB244R (Lactobacillus plantarum LB244R, deposited as DSM32996)

[0230] - LB349R (Leuconostoc mesenteroides LB349R, deposited as DSM 33093)

[0231] - LB10G (Weissella viridis LB10G, deposited as DSM 32906).

[0232] Thus, one aspect of the present invention relates to novel isolated bacterial strains and / or compositions comprising one or more bacterial strains having antimicrobial activity against pathogenic Propionibacterium acnes.

[0233] Another aspect of the present invention relates to a novel isolated bacterial strain having growth inhibitory activity against pathogenic Propionibacterium acnes.

[0234] Another aspect of the present invention relates to novel isolated bacterial strains capable of co-aggregating with pathogenic P. acnes.

[0235] Another aspect of the invention relates to a composition comprising a bacterial strain having antimicrobial activity and / or growth inhibitory activity and / or coaggregation activity against pathogenic Propionibacterium acnes.

[0236] Another aspect of the present invention is to provide the use of a composition comprising a bacterial strain having antimicrobial activity and / or growth inhibitory activity and / or co-aggregation activity against pathogenic Propionibacterium acnes.

[0237] Another aspect of the present invention is to provide a composition for treating an infection caused by Propionibacterium acnes.

[0238] Deposit of biological materials

[0239] The following biological materials, microorganisms, have been deposited at the German Collection for Microorganisms and Cell Cultures:

[0240] Lactobacillus plantarum LB356R, deposited as DSM33094

[0241] Lactobacillus plantarum LB244R, deposited as DSM32996

[0242] Leuconostoc mesenteroides LB349R, deposited as DSM 33093

[0243] Weissella viridis LB10G, deposited as DSM32906.

[0244] It should be noted that embodiments and features described in the context of one aspect of the invention may also be applicable to other aspects of the invention.

[0245] All patent and non-patent references cited in this application are incorporated herein by reference in their entirety.

[0246] The invention will now be described in more detail in the following non-limiting examples.

[0247] Example

[0248] Example 1 Strain screening and identification

[0249] sample

[0250] In order to identify and select bacterial strains according to the present invention, a collection of lactic acid bacteria (LAB) strains was established. Samples from different sources, such as homemade sauerkraut, kimchi and healthy human donor samples (vaginal, oral, anal, skin) were collected to isolate at least 995 lactic acid bacteria. Samples were collected on Man Rogosa Sharp (MRS, Sigma-Aldrich) broth and agar medium and cultured anaerobically at 37°C overnight or until colonies formed. The isolates were plated and subcultured until pure colonies were obtained. Pure colonies were stored in MRS broth containing 25% glycerol at -80°C for future use. The strains were identified using standard methods using 16S rRNA Sanger sequencing.

[0251] Example 2 - Co-aggregation

[0252] P. acnes strains obtained from BEI resources were used as research test organisms for screening.

[0253]

[0254] Propionibacterium acnes ATCC 6919 (America Type Culture Collection) was used as a typing strain of Propionibacterium acnes.

[0255] Lactobacillus rhamnosus LGG (Chr. Hansen) was used as a control probiotic strain in all experiments, and the YUN product ACN+ cream containing live lactic acid bacteria (YUN Probiotherapy, online shop: www.shop.yun.be) was used as a control for commercially available probiotic lactic acid bacteria for topical use. Lactobacillus rhamnosus LBB and YUN strains were grown as LAB on MRS as described in the examples.

[0256] Coaggregation was determined according to the known method Cisar, JO et al. (1979) "Specificity of Coaggregation Reactions between Human Oral Streptococci and Strains of Actinomyces Viscosusor Actinomyces Naeslundii" Infection and Immunity 24 (3): 742-52. Inocula of all lactic acid bacteria (LAB) were grown anaerobically overnight in MRS broth, and P. acnes strains were grown anaerobically in BHI broth at 37°C to a cell density of approximately 10 8CFU / ml (2-3 days). Overnight cell samples were harvested by centrifugation (6000 rpm, 2 minutes) and the supernatant was removed from the pellet. The pellet was washed twice in 1X PBS buffer.

[0257] Resuspend the cell pellet in 1XPBS and aliquot 500µl of P. acnes and LAB strains into 24-well plates. Incubate the plates on a shaker (200rpm). Visually observe the formation of self-aggregates and co-aggregates after 1, 2, 3 and 24 hours.

[0258] As a control for auto-aggregation, each strain was mixed with PBS in a final volume of 1500 μl per well.

[0259] Co-aggregation was determined by mixing P. acnes and LAB strains. Each strain was suspended in PBS buffer and mixed 1:1 in a final volume of 1500ul per well.

[0260] The plates were incubated on a shaker at approximately 200 rpm for 24 hours. Coaggregate formation was observed after 1 hour, 2 hours, 3 hours and 24 hours.

[0261] The formation of coaggregates was scored visually using the following scale from 1-5:

[0262] 1: No aggregation

[0263] 2: Visualize initial aggregation

[0264] 3: Aggregate formation <0.5mm

[0265] 4: Aggregate formation > 0.5 mm and < 1 mm

[0266] 5: Aggregate formation > 1 mm

[0267] Table 1: Coaggregation was measured using visual assessment from 1 to 5 (see scale above). Data are shown for 1 hour and 24 hours of incubation

[0268]

[0269]

[0270] Thirty-four LAB strains were identified as being able to coaggregate with pathogenic P. acnes strains, and the coaggregates of four strains (LB10G, LB244R, LB349R, and LB356R) are listed in Table 1 , which coaggregated relatively more with pathogenic ribotypes (RT4, RT5, and RT8) of P. acnes than with the nonpathogenic ribotype (RT6).

[0271] ATCC-type strains generally perform slightly differently in testing; they are also easier to grow and can be grown anaerobically overnight in BHI to an OD(600) > 1.

[0272] The control LGG and strains in the commercial product from YUN did not show any ability to co-aggregate with P. acnes.

[0273] Coaggregation can inhibit the growth of the organism and affect the ability to produce biofilm. Therefore, LABs that are able to coaggregate with P. acnes may inhibit biofilm formation and reduce the virulence of acne strains.

[0274] Example 3 - Effect of pH on coaggregation.

[0275] Coaggregation was determined according to the known method Cisar, JO et al. (1979) as described above.

[0276] All lactic acid bacteria (LAB) inocula were grown anaerobically overnight in MRS broth, and P. acnes strains were grown anaerobically in BHI broth at 37°C to a cell density of approximately 10 8 CFU / ml (2-3 days). Overnight cell samples were harvested by centrifugation (6000 rpm, 2 minutes) and the supernatant was removed from the pellet. The beads were washed twice in the pH adjusted buffer used for the coaggregation assay.

[0277] MOPS (3-(N-morpholino)propanesulfonic acid) obtained from Merck (product 69947) and MES (2-(N-morpholino)ethanesulfonic acid) buffer obtained from Sigma (product M2933) were used at a concentration of 100 mM. MES was used at pH 4.5, pH 5, pH 5.5, pH 6, and pH 6.5. MOPS was used at pH 6.5, pH 7, pH 7.5, and pH 8.

[0278] It was found that coaggregation was slightly less in the inert buffers MES and MOPS compared to PBS buffer. MES and MOPS allowed the evaluation of the effect of pH on coaggregation over a wide pH range, limiting any effect of the buffer.

[0279] The cell pellet of each strain was resuspended in each buffer and 500 µl of P. acnes and LAB strains were aliquoted into 24-well plates with each pH value. The plates were incubated on a shaker (200 rpm). The formation of self-aggregates and co-aggregates was visually observed after 1, 2, 3, and 24 hours.

[0280] As a control for auto-aggregation, each strain at each pH value was included in a final volume of 1000 ul in the well. No significant differences in auto-aggregation were found at different pH values. No significant auto-aggregation was observed for any of the strains tested.

[0281] Co-aggregation was determined by mixing P. acnes and LAB strains. Each strain was suspended in PBS buffer and mixed 1:1 in a final volume of 1000ul per well.

[0282] The plates were incubated on a shaker at approximately 200 rpm for 24 hours. Coaggregate formation was observed after 1 hour, 2 hours, 3 hours and 24 hours.

[0283] The formation of coaggregates was scored visually using the following scale from 1-5:

[0284] 1: No aggregation

[0285] 2: Visualize initial aggregation

[0286] 3: Aggregate formation <0.5mm

[0287] 4: Aggregate formation > 0.5 mm and < 1 mm

[0288] 5: Aggregate formation > 1 mm

[0289] Table 2: Coaggregation was measured using a visual assessment from 1 to 5 (see scale above). Data are shown for 1 hour incubation and 2 strains LB244R and LB356R as well as pathogenic P. acnes HM-513 and non-pathogenic P. acnes HM-554.

[0290]

[0291] For strains that were able to coaggregate, the results were generally the same. A decrease in pH resulted in increased coaggregation. All four strains, LB244R, LB356R, LB10G, and LB349R, resulted in increased coaggregation when the pH was lowered. The ratio between coaggregation with pathogenic and nonpathogenic P. acnes was constant and did not appear to be significantly affected by changes in pH. Thus, at low pH, the strains still coaggregated more with pathogenic P. acnes than with nonpathogenic P. acnes.

[0292] The control LGG and strains in the commercial product from YUN did not show any ability to coaggregate with P. acnes in any buffer.

[0293] Coaggregation can inhibit the growth of organisms and affect the ability to produce biofilm. Therefore, LABs that are able to coaggregate with P. acnes can inhibit biofilm formation and reduce the virulence of acne strains. Therefore, this effect is expected to be further improved at pH values ​​below 7.

[0294] Example 4 - Spot Detection on Lawn

[0295] Field trials were conducted on turfgrass to test growth inhibition and antimicrobial metabolites using Zhang P. et al. (2015) Interstrain interactions between bacteriaisolated from vacuum-packaged refrigerated beef. Appl Environ Microbiol 81:2753–2761. doi:10.1128 / AEM.03933-14 and Arena, MP et al. (2016) Use of Lactobacillus plantarum strains as a biocontrol strategy against foodborne pathogenic microorganisms. Frontiers in Microbiology 7 (APR): 1-10. https: / / doi.org / 10.3389 / fmicb.2016.00464.

[0296] Bacterial strains from Example 1 were cultured from stock samples into 2 mL of MRS broth in 24-well plates. Each P. acnes test strain was cultured anaerobically in BHI broth in approximately 200 mL conical flasks for 48-72 hours (OD600>1). LAB isolates were grown overnight at 37°C. The cell density of the P. acnes overnight culture was adjusted to an optical density at 600 nm (OD600) of 1 in BHI broth and then diluted to 10 in PBS buffer. -2 dilution. 200 µL of the cell suspension was spread on a BHI agar plate. The plate with the P. acnes lawn was placed to dry in sterile air for approximately 10-20 minutes. Three aliquots of 20 µL of the isolated LAB were spotted on the P. acnes lawn. The plates were air dried and then incubated anaerobically at 37°C for 48 hours. The inhibition zone was measured in mm as the clearing area around the spot and the diameter was measured as the full diameter of the spot. Growth inhibition was observed because the LAB was able to overgrow the P. acnes strain in the area of ​​the spot indicated by (+) in Table 2. No growth inhibition was detected if the P. acnes strain was able to outgrow the bacterial strain in the spot as indicated by (-) in Table 2.

[0297] 15 strains were able to outcompete all P. acnes strains, and 5 LAB strains were identified to have significant growth inhibition against P. acnes with inhibition zones exceeding 1 mm around the spots.

[0298] Lactobacillus rhamnosus LGG (Chr. Hansen) and Yun product were used as control commercial probiotic strains. Neither Lactobacillus rhamnosus nor YUN strains were able to inhibit the growth of any pathogenic P. acnes test strains, nor did Lactobacillus rhamnosus LGG or YUN product produce any clearance zones.

[0299] P. acnes ATCC 6919 appeared to be more susceptible to growth inhibition, and many strains were identified as having growth inhibitory activity against ATCC 6919 but did not appear to have any growth inhibitory activity against pathogenic isolates (RT4, RT5, and RT8) or even against nonpathogenic isolates (RT6).

[0300] Table 3:

[0301] Spot assay targeting the type strain (Propionibacterium acnes ATCC 6919), with the inhibitor zone measured for each strain having an effect.

[0302] *The diameter is the average of three measurements of the inhibition zone. **The length of the inhibition zone is the average of three measurements from the edge of the bacterial spot to the growth of P. acnes.

[0303]

[0304] Table 4 Inhibitor zone measured for each strain with effect in spot test with HM-523 as target strain. *The diameter is the average of three measurements of the inhibition zone. **The length of the inhibitor zone is the average of three measurements from the periphery of the bacterial spot to the growth of HM-523.

[0305]

[0306] Table 5: Growth inhibition and antimicrobial metabolites. The inhibition zone was determined as the mean of three measurements.

[0307]

[0308] Example 5 - Evaluation of bacterial lysates.

[0309] Bacterial lysates were generated using 3 different methods.

[0310] LAB isolate (LB356R) was grown overnight at 37°C in MRS.

[0311] Lysis 1: Place a tube with 15 ml of overnight culture on ice and lyse the cells by sonication for 30 min using a Q125 sonicator (QSonica).

[0312] Lysate 2: 15 ml of overnight culture was adjusted to pH 3 with 0.1 M HCl and incubated at 45 degrees Celsius for 2 days.

[0313] Lysate 3: 15 ml of overnight culture was supplemented with 5% SDS (Sigma-Aldrich L3771) and stored at -20°C for 1 hour and incubated at 50°C for 1 hour. The cycle of changing the temperature from -20°C to +50°C was performed 2 × 8 hours.

[0314] Viable cells were assessed for each lysate after lysate treatment.

[0315] The lysates were evaluated for their ability to maintain activity against P. acnes HM-512 by testing the ability to coaggregate as described in Example 3 and also by co-incubating each lysate with BHI inoculated with HM-512 for approximately 10 4 Growth inhibitory activity was assessed by measuring CFU / ml and growth and growth inhibition were followed by measuring OD (600 nm) using a spectrophotometer.

[0316] All three lysates were shown to inhibit the growth of HM-512. Lysates 1 and 2 were found to co-aggregate comparable to live LB356R cells, while lysate 3 was found to co-aggregate less than live cells with a co-aggregation score of 3 using the scale in Example 2.

[0317] Example 6

[0318] The strain LAB strain LB356R was formulated in a vegetable oil mixture containing jojoba oil and almond oil (1:1). 8 The strains were added at a concentration of CFU / ml oil.

[0319] Ten adolescents (15-19 years) with acne-prone skin were treated with the composition once daily. Visual assessment of acne severity was performed before treatment ( Figure 1a ) and after treatment ( Figure 1b ) were determined by taking pictures of the . Visual improvement was observed in all test subjects and, on average, improvement was detectable after 3-4 days of treatment.

[0320] Example 7:

[0321] The anti-inflammatory activity of LAB cells was determined on a 3D tissue model of reconstructed human epidermis (RHE) manufactured by Episkin (Lyon, France) (Batch 19-RHE-164). RHE cells were treated with each LAB strain. Overnight cultures of each strain in MRS were diluted in saline to OD = 0.8 and incubated with RHE cells for 4 h. Gene expression was determined by qRT-PCR for TSLP, TNF-a, IL-8, TLR-2, and HBD2. The integrity of the RNA was assessed by loading the extracted RNA on a 1% agarose gel: ribosomal bands 18S and 28s were detected. The High Capacity cDNA Reverse Transcription Kit was used to synthesize cDNA from RNA. For real-time PCR, an Applied Biosystems 7500 Fast Real-Time Fluorescence Quantitative Taqman PCR Assay was used. GAPDH was used as an endogenous control gene to normalize the input amount. Significant HBD2 upregulation was observed for all 4 LAB strains. Probiotic strains can effectively enhance the host's innate immune defense against infection.

[0322] Example 8:

[0323] Biofilm formation is determined as the ability of microorganisms to begin to bind to and grow on a surface.

[0324] All lactic acid bacteria (LAB) inocula were grown anaerobically overnight in MRS broth, and P. acnes strains were grown anaerobically in BHI broth at 37°C to a cell density of approximately 10 8 CFU / ml (2-3 days). Overnight cell samples were harvested by centrifugation (6000 rpm, 2 minutes) and the supernatant was removed from the pellet. The pellet was washed twice in 1X PBS buffer.

[0325] The cell pellet was resuspended in BHI medium and diluted to a cell concentration of approximately 10 6 CFU / ml. 100 µL of P. acnes and 100 µL of LAB strains were aliquoted into a 96-well plate (Sigma cell culture plate SIAL0596-50EA, flat bottom). The plate was incubated at 37°C for 1 hour to allow the cells to adhere to the bottom surface of the wells. The microtiter plate was emptied from the culture medium and the bottom of the wells were washed once with PBS.

[0326] 200 µL of sterile BHI medium was added to each well, and the level of surface-attached cells was determined by absorbance measurement using a standard curve for each strain, and the correlation between OD measurements (detected at 600 nm) and cell numbers was determined as CFU / ml by plate counting and calculated as the number of attached viable CFU / well.

[0327] Initial early biofilm formation was followed by microscopy and image analysis using an oCelluScope from BioSense Solutions, Denmark. Biofilm growth was observed for 4 hours at 37°C (anaerobic conditions). Biofilm growth was found to be directly correlated with the number of surface-attached cells.

[0328] Controls included a selective medium (MRS) for LAB to reduce the contribution of LAB to the biofilm, as well as a control for monoculture biofilm formation of each P. acnes strain. oCelluScope allowed for visual and image analysis to assess the dominant strain in the biofilm based on cell morphology.

[0329] Table 6: Initial attachment to the surface (number of cells attached to the bottom of the wells, numbers shown are the average of triplicate measurements):

[0330]

[0331] It was observed that non-pathogenic strains (HM-554 and HM-555) were generally less attached to the surface compared to pathogenic strains (HM-523, HM-513, and HM-512). When the probiotic strains were not present, the initial attachment level of the non-pathogenic strains was approximately 10 to 500 cells per well. Whereas, when the probiotic strains were not present, the initial attachment of the pathogenic strains was approximately 1000 to 10,000 cells per well. The presence of LB10G, LB244R, LB349R, and LB356R significantly inhibited the initial attachment of the pathogenic strains. Thus, a 2 log unit reduction in initial attachment was observed for these 4 probiotic strains, and this reduction was the only significant change measured. No effect was observed for the two control LAB strains (LGG and YUN) in preventing initial attachment to the surface.

[0332] References

[0333] Fitz-Gibbon et al.2013, J. Invest.Dermatol.133:2152-60.doi:10.1038 / jid.2013.21

[0334] McDowell et al.2013, PLoS ONE 8(9): e70897

[0335] Dreno et al.2018, JEADV 32(suppl. 2), 5-14. doi:10.1111 / jdv.15043

[0336] WO 2017 / 220525.

Claims

1. A composition comprising one or more bacterial strains selected from the following for use in the preparation of a composition for inhibiting pathogenic Propionibacterium acnes ( C. acnes ) Use of topical compositions of strains: Lactobacillus plantarum LB356R, deposited as DSM33094; Lactobacillus plantarum LB244R, deposited as DSM32996; Leuconostoc mesenteroides LB349A, deposited as DSM33093; and Weissella viridis LB10G, deposited as DSM32906; in, Nonpathogenic Propionibacterium acnes ( C. acnes ) strains, the one or more bacterial strains are effective against pathogenic Propionibacterium acnes ( C. acnes ) strains showed stronger inhibitory effects.

2. Use of a composition comprising one or more bacterial strains selected from the group consisting of: Lactobacillus plantarum LB356R, deposited as DSM33094; Lactobacillus plantarum LB244R, deposited as DSM32996; Leuconostoc mesenteroides LB349A, deposited as DSM33093; and Weissella viridis LB10G, deposited as DSM32906; in, Nonpathogenic Propionibacterium acnes ( C. acnes ) strains, the one or more bacterial strains are effective against pathogenic Propionibacterium acnes ( C. acnes ) strains showed stronger inhibitory effects.

3. The use according to claim 1 or 2, wherein the inhibition is growth inhibition, co-aggregation inhibition, biofilm inhibition or a combination thereof of the P. acnes strain.

4. The use according to claim 1 or 2, wherein The inhibition of pathogenic P. acnes strains was 10% or more stronger relative to the inhibition of non-pathogenic P. acnes strains.

5. The use according to claim 1 or 2, wherein The inhibition of pathogenic P. acnes strains was 50% or more potent relative to the inhibition of non-pathogenic P. acnes strains.

6. The use according to claim 1 or 2, wherein The inhibition of pathogenic P. acnes strains was 100% or more potent relative to the inhibition of non-pathogenic P. acnes strains.

7. The use according to claim 1 or 2, wherein The inhibition of pathogenic P. acnes strains was 200% or more potent relative to the inhibition of non-pathogenic P. acnes strains.

8. The use according to claim 1 or 2, wherein the pathogenic Propionibacterium acnes strain is selected from the group consisting of Propionibacterium acnes ribotype RT4; ribotype RT5; and ribotype RT8.

9. The use according to claim 1 or 2, wherein the one or more bacterial strains are provided as one or more live strains.

10. The use according to claim 9, wherein the concentration of one or more bacterial strains is 10 3 Up to 10 14 The number of colony forming units (CFU) was within the range of 1.

11. The use according to claim 9, wherein the concentration of one or more bacterial strains is 10 7 -10 11 CFU range.

12. The use according to claim 1 or 2, wherein the composition is for topical application and is formulated as a paste, powder, lotion, foam or ointment.

13. Use according to claim 1 or 2, wherein the composition is for topical application and is formulated into talc, custard, cream, oil or serum.

14. The use according to claim 1 or 2, wherein the pH of the composition is below pH 6.

5.

15. The use according to claim 1 or 2, wherein the pH of the composition is below pH 5.

16. The use according to claim 1 or 2, wherein the pH of the composition is in the range of pH 4-6.5.

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