Novel skin care compositions

Abstract

The present invention relates to novel skin care compositions. Specifically, the present invention relates to a cosmetic or therapeutic skin care composition comprising a combination of live bacteria of at least one Propionibacterium acnes strain and an emollient that specifically supports the viability of said bacteria during storage and / or their ability to replicate upon application to the skin. Preferably, the cosmetic or therapeutic skin care composition comprises a bacterium of at least one Propionibacterium acnes strain selected from the group consisting of D1, A5, C1, C3, H1, H2, H3, K1, K2, K4, K6, K8, K9, L1 and F4. The invention also provides a method of treating or preventing acne by applying a skin care composition of the invention to an area of skin in need of treatment. The invention also relates to the use of the skin care composition of the invention for the treatment or prevention of acne.

Description

[0001] This application is a divisional application of an application with an international filing date of December 18, 2019, international application number PCT / EP2019 / 085875, which entered the Chinese national phase on April 1, 2022, application number 201980101000.1, and invention title "New Skin Care Composition". Technical Field

[0002] The present invention generally relates to the field of skin care. More specifically, the present invention relates to a cosmetic or therapeutic skin care composition comprising a combination of live bacteria of at least one Cutibacterium acnes (C. acnes) strain and an emollient that specifically supports the viability of the strain during storage and / or their ability to replicate after application to the skin. Preferably, the cosmetic or therapeutic skin care composition comprises bacteria of at least one Cutibacterium acnes strain selected from D1, A5, C1, C3, H1, H2, H3, K1, K2, K4, K6, K8, K9, L1 and F4. The present invention also provides a method for treating or preventing acne by applying the skin care composition of the present invention to a skin area in need of treatment. The present invention also relates to the use of the skin care composition of the present invention for treating or preventing acne. Background Art

[0003] Acne vulgaris is a widespread long-term skin disorder that affects more than 600 million people worldwide. Acne is most common among teenagers, although it affects people of all ages. It is usually caused by a combination of sebaceous gland hyperplasia, sebum overproduction, and impaired keratinization. As a result, hair follicles become clogged with oil and dead skin cells, leading to comedones and oily skin (Pschyrembel, Klinisches Wörterbuch, 258. ed., Walter de Gruyter-Verlag, Berlin, 1998). Colonization of the affected skin areas by bacteria may additionally cause inflammation. Acne mainly affects skin areas with a large number of sebaceous glands, especially the face, upper chest, and back. The occurrence of acne can lead to emotional distress and mental problems, such as reduced self-esteem and depression.

[0004] Oily skin is a transitional state between healthy skin and acne-prone skin. In oily skin, the sebaceous glands of the skin produce an excess of sebum, which then serves as an ideal nutrient for many bacteria and yeasts, including different species of the anaerobic Gram-positive bacterium Propionibacterium acnes (formerly known as Propionibacterium acnes) and yeasts of the genus Pityrosporum. These microorganisms break down sebum into glycerol and fatty acids, thereby further inducing sebum production in the sebaceous glands and damaging the hair follicle walls in the skin. This results in skin inflammation and the formation of comedones, pustules, nodules, and cysts, which typically form scars upon healing, thus permanently affecting the optical appearance of the subject's skin (W. Umbach [Ed.], Kosmetik, Entwicklung, Herstellung und Anwendung kosmetischer Mittel, 2. Edition Thieme Verlag, Stuttgart, 1995).

[0005] There appear to be many different factors contributing to the development of acne, including genetics, hormonal status, stress, and diet. In addition, the anaerobic bacterial species Propionibacterium acnes (formerly Propionibacterium acnes) is thought to play an important role in the development of acne, as large numbers of these bacteria are frequently found in patients with moderate or severe inflammatory acne. However, the underlying mechanism is not fully understood.

[0006] Currently, standard treatment of acne typically involves topical application of antibiotics including erythromycin, clindamycin, metronidazole, sulfacetamide, doxycycline, or minocycline to reduce the number of bacteria, particularly Propionibacterium acnes. However, some of these antibiotics exhibit considerable side effects, making their use inconvenient for patients. In addition, treatment of acne with antibiotics is associated with a high recurrence rate, due to the fact that small numbers of Propionibacterium acnes survive and resume growth after termination of antibiotic treatment. Thus, there is a need for new methods of treating or preventing acne that are substantially free of side effects and provide long-lasting effects.

[0007] In more recent treatment approaches, acne has been considered the result of a distortion of the human skin microbiome caused by specific strains of Propionibacterium acnes (Holmes, 2013; Lomholt and Kilian, 2010). Researchers have only recently begun to study the skin microbiome (Belkaid and Segre, 2014; Oh et al., 2014). Although the skin is colonized by a large number of harmless or even beneficial microorganisms (Grice and Segre, 2011), alterations in the microbiome can cause diseases such as acne (Bek-Thomsen et al., 2008; Holmes, 2013; Kong et al., 2012; Fitz-Gibbon et al., 2013). Such a distortion may be caused by specific subsets of the skin bacterium Propionibacterium acnes (Lomholt and Kilian, 2010). Therefore, it has been proposed to modulate the skin microbiome in an attempt to restore a healthy microbiome. For example, WO 2016 / 172196 A1 discloses a method of treating acne in a subject, the method comprising first administering a disinfectant or an antibiotic to the skin of the subject, and subsequently administering a composition comprising one or more live strains of Propionibacterium acnes. Similarly, WO 2018 / 073651 A1 discloses a composition for treating acne, which comprises two or more strains of Propionibacterium acnes, including Propionibacterium acnes strain C3 and / or K8. SUMMARY OF THE INVENTION

[0008] In a first aspect, the present invention relates to a skin care composition for topical administration to the skin, comprising:

[0009] (a) freeze-dried or spray-dried live bacteria of at least one strain of Propionibacterium acnes (C. acnes); and

[0010] (b) an emollient selected from dicaprylyl carbonate, ethylhexyl cocoate, and mixtures thereof.

[0011] In certain embodiments, the skin care composition comprises dicaprylyl carbonate as the emollient. In certain embodiments, dicaprylyl carbonate is present in the skin care composition in an amount of 0.05 to 25.0% (w / w). In certain embodiments, dicaprylyl carbonate is present in the skin care composition in an amount of 7.5 to 10.0% (w / w).

[0012] In certain embodiments, the skin care composition comprises ethylhexyl cocoate as the emollient. In certain embodiments, ethylhexyl cocoate is present in the skin care composition in an amount of 0.05 to 25.0% (w / w). In certain embodiments, ethylhexyl cocoate is present in the skin care composition in an amount of 7.5 to 10.0% (w / w).

[0013] In certain embodiments, the skin care composition comprises a mixture of dicaprylyl carbonate and ethylhexyl cocoate as an emollient.

[0014] In certain embodiments, the skin care composition comprises equal amounts of dicaprylyl carbonate and ethylhexyl cocoate.

[0015] In certain embodiments, the total amount of emollient in the composition does not exceed 20.0% (w / w).

[0016] In certain embodiments, the skin care composition does not contain any other emollient other than dicaprylyl carbonate and / or ethylhexyl cocoate.

[0017] In certain embodiments, the skin care composition further comprises a thickening agent selected from Chondrus crispus extract, hydroxypropyl starch phosphate, and mixtures thereof.

[0018] In certain embodiments, the skin care composition further comprises a filler selected from distarch phosphate, tapioca starch, and mixtures thereof.

[0019] In certain embodiments, the skin care composition further comprises an antioxidant selected from tocopherol, tocopheryl acetate, and mixtures thereof.

[0020] In certain embodiments, the skin care composition further comprises a preservative selected from ethanol, phenoxyethanol, octanediol, methylpropanediol, and mixtures thereof.

[0021] In certain embodiments, the skin care composition further comprises PEG-40 hydrogenated castor oil as a solubilizer.

[0022] In certain embodiments, the skin care composition further comprises a citric acid / citrate buffer as a pH regulator.

[0023] In certain embodiments, the skin care composition comprises at least one Propionibacterium acnes strain selected from the single-locus sequence typing (SLST) types D1, A5, C1, C3, H1, H2, H3, K1, K2, K4, K6, K8, K9, L1, and F4. In certain embodiments, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain. In certain embodiments, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes SLST K8 strain. In certain embodiments, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain.

[0024] In certain embodiments, the skin care composition further comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes SLST A5 strain. In certain embodiments, the skin care composition further comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes SLST F4 strain.

[0025] In certain embodiments, the concentration of each Propionibacterium acnes strain is at least 0.5% (w / v) of the skin care composition. In certain embodiments, the at least one Propionibacterium acnes SLST C3 strain and the at least one Propionibacterium acnes SLST K8 strain have approximately equal concentrations in the composition. In certain embodiments, the at least one Propionibacterium acnes SLST C3 strain is present in a higher concentration than the at least one Propionibacterium acnes SLST K8 strain in the composition. In other embodiments, the at least one Propionibacterium acnes SLST K8 strain is present in a higher concentration than the at least one Propionibacterium acnes SLST C3 strain in the composition.

[0026] In certain embodiments, each Propionibacterium acnes strain in the composition is present in an amount of 10 4 -10 11 colony forming units / ml (CFU / ml), preferably 10 7 -10 10 CFU / ml. In certain embodiments, the total amount of bacteria in the composition is 10 4 -10 11 CFU / ml, preferably 10 7 -10 10 CFU / ml.

[0027] In certain embodiments, the skin care composition takes the form of a gel, cream, ointment or lotion.

[0028] In a second aspect, the present invention relates to a method of improving the appearance of the skin of a subject and / or regulating sebum production of the skin cells of the subject and / or maintaining healthy skin of the subject, the method comprising topically administering the skin care composition of the present invention. In certain embodiments, the subject is human.

[0029] In a third aspect, the present invention relates to a method of treating or preventing a disorder selected from acne, oily skin, progressive macular hypomelanosis, dandruff, atopic eczema, atopic dermatitis and rosacea in a subject, the method comprising topically administering the skin care composition of the present invention. In certain embodiments, the subject is human. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 Shows the composition of prototype formulations 1-15 used in the reactivation experiments using lyophilized bacteria.

[0031] Figure 2 Shows the results of experiments analyzing the reactivation of lyophilized bacteria from prototype formulations 1-11.

[0032] Figure 3 Shows the results of experiments analyzing the reactivation of lyophilized bacteria from prototype formulations 12-15.

[0033] Figure 4 Shows the composition of additional prototype formulations 1-11 used in the reactivation experiments using lyophilized bacteria.

[0034] Figure 5 Shows the results of experiments analyzing the reactivation of lyophilized bacteria from the additional prototype formulations 1-11.

[0035] Figure 6 Shows the results of experiments analyzing the reactivation of lyophilized bacteria from comparative formulations V1-V10.

[0036] Figure 7 Shows the composition of comparative formulations V1-V10 used in the reactivation experiments using lyophilized bacteria. DETAILED DESCRIPTION

[0037] Although these methods and compositions provide new and efficient modulation of the microbiome to improve or eliminate the symptoms of acne, it has been found that providing live bacteria in a cosmetic skin care composition presents significant problems. For example, some of the components commonly used in commercial skin care products interfere with the viability of bacteria. When these compounds are combined with lyophilized or spray-dried bacteria into a composition, they may interfere with the viability of the bacteria or have a negative impact on the growth rate of the bacteria after application to the skin. Accordingly, there is a need for a new skin care product that is formulated in a manner compatible with the application of lyophilized or spray-dried bacteria and that does not inhibit the growth of these bacteria after the product is applied to the skin.

[0038] It has now surprisingly been found that the viability of lyophilized or spray-dried bacteria, particularly bacterial strains of the species Propionibacterium acnes, in a skin care composition during storage and their ability to grow after application to the skin can be retained or significantly improved by excipients added to the final skin care product formulation. The present invention is based on the identification of a group of excipients for skin care compositions that do not significantly interfere with or can improve the viability and reactivation of growth of bacteria. These compounds have been found to be highly compatible with the administration of bacteria, particularly bacterial strains of the species Propionibacterium acnes.

[0039] Accordingly, in a first aspect, the present invention relates to a skin care composition for topical administration to the skin, the composition comprising, consisting essentially of, or consisting of the following components:

[0040] (a) at least one lyophilized or spray-dried live bacterium of a Propionibacterium acnes strain; and

[0041] (b) an emollient selected from dicaprylyl carbonate, ethylhexyl cocoate, and mixtures thereof.

[0042] In the context of the present invention, lyophilized or spray-dried live bacteria are used in the skin care composition. This means subjecting the live bacteria to a drying process that maintains their viability but minimizes their metabolic processes. In lyophilized or spray-dried form, the bacteria can be stored for months or even years. Once they are applied to the skin, such as human skin, the metabolism of the bacteria is reactivated, enabling them to resume growth. They multiply on the skin surface and replace pathogenic bacterial strains, thereby restoring a diverse, healthy, and balanced skin microbiome.

[0043] In one embodiment, the live Propionibacterium acnes bacteria are in spray-dried form. The principle of spray drying is based on dispersing a solution into fine droplets and introducing the droplets into a hot air stream. The solvent evaporates from the substrate droplets, leaving behind agglomerates of the dried product. Standard spray drying equipment can be used, such as the Mini Spray Dryer B-290 from Büchi Labortechnik GmbH (Essen, Germany) or the Mobile Minor™ Spray Dryer from GEA (Berlin, Germany).

[0044] In one embodiment, the live Propionibacterium acnes bacteria are in freeze-dried or lyophilized form. Freeze-drying or lyophilization is a process that includes freezing the product, reducing the pressure, and heating to allow the frozen water in the material to sublime. Various different methods can be used to freeze the product. For example, freezing can be achieved by using a standard refrigerator or a freezing bath. Cooling the product below its triple point ensures that sublimation occurs upon heating. To prevent the formation of large crystals that could damage the structure of the product to be dried, the freezing is carried out rapidly. When the frozen water sublimes, approximately 95% of the water in the product is removed. Most materials can be dried to a residual moisture content of 1 - 5%. Standard freeze-drying equipment can be used, such as the Lyovac™ equipment from GEA (Berlin, Germany), the Gamma 2-20 Freeze Dryer LCM-1 from Christ (Osterode am Harz, Germany), or the Christ Martin™ Alpha 1-2 Lyophilizer from Fisher Scientific GmbH (Schwerte, Germany).

[0045] One or more excipients that do not interfere with the viability and growth reactivation of the live bacteria in the freeze-drying or spray drying are selected from emollients, thickeners, fillers, antioxidants, preservatives, solubilizers, and pH regulators. Suitable members of each of these compound groups will be discussed below.

[0046] Emollient

[0047] According to the present invention, the skin care composition comprises an emollient in combination with the freeze-dried or spray-dried live bacteria of the at least one Propionibacterium acnes strain. As used herein, an emollient is a compound that moistens and / or softens the skin. Emollients generally reduce skin roughness, cracking, and / or irritation by penetrating deeper into the skin. Emollients commonly used in skin care products include vegetable oils such as sesame oil, coconut oil, olive oil, almond oil, macadamia nut oil, cottonseed oil, or peanut oil, silicone oils such as dimethicone and cyclomethicone, fatty acids, and fatty alcohol ethers.

[0048] According to the present invention, the skin care composition comprising a freeze-dried or spray-dried live bacterium of at least one Propionibacterium acnes strain comprises an emollient selected from dicaprylyl carbonate, ethylhexyl cocoate and mixtures thereof. In a preferred embodiment, the skin care composition comprises a freeze-dried or spray-dried live bacterium of at least one Propionibacterium acnes strain and dicaprylyl carbonate as an emollient. In another preferred embodiment, the skin care composition comprises a freeze-dried or spray-dried live bacterium of at least one Propionibacterium acnes strain and ethylhexyl cocoate as an emollient. In yet another preferred embodiment, the skin care composition comprises a freeze-dried or spray-dried live bacterium of at least one Propionibacterium acnes strain and both dicaprylyl carbonate and ethylhexyl cocoate as emollients. Particularly preferably, the skin care composition does not contain any other emollients other than dicaprylyl carbonate and / or ethylhexyl cocoate.

[0049] When dicaprylyl carbonate is used as an emollient, it is preferably used in an amount of 0.05 to 25.0% (w / w), more preferably 2.0 to 20.0% (w / w), more preferably 5.0 to 10.0% (w / w) or 7.5 to 10.0% (w / w) in the final skin care composition. In other words, the amount of dicaprylyl carbonate in the skin care composition of the present invention can be at least 0.05% (w / w), at least 0.1% (w / w), at least 0.25% (w / w), at least 0.5% (w / w), at least 0.75% (w / w), at least 1.0% (w / w), at least 1.25% (w / w), at least 1.5% (w / w), at least 1.75% (w / w), at least 2.0% (w / w), at least 2.5% (w / w), at least 3.0% (w / w), at least 4.0% (w / w), at least 5.0% (w / w), at least 6.0% (w / w), at least 7.0% (w / w), at least 8.0% (w / w) or at least 9.0% (w / w).

[0050] When using ethylhexyl cocoate as an emollient, it is preferably used in the final skin care composition in an amount of 0.05 to 25.0% (w / w), more preferably 1.0 to 10.0% (w / w), even more preferably 5.0 to 10.0% (w / w) or 7.5 to 10.0% (w / w). In other words, the amount of ethylhexyl cocoate in the skin care composition of the present invention can be at least 0.05% (w / w), at least 0.1% (w / w), at least 0.25% (w / w), at least 0.5% (w / w), at least 0.75% (w / w), at least 1.0% (w / w), at least 1.25% (w / w), at least 1.5% (w / w), at least 1.75% (w / w), at least 2.0% (w / w), at least 2.5% (w / w), at least 3.0% (w / w), at least 4.0% (w / w), at least 5.0% (w / w), at least 6.0% (w / w), at least 7.0% (w / w), at least 8.0% (w / w) or at least 9.0% (w / w).

[0051] When dicaprylyl carbonate and ethylhexyl cocoate are used in combination with each other as emollients, preferably the total amount of the emollients is at least 0.05% (w / w), but does not exceed 20.0% (w / w), more preferably does not exceed 15.0% (w / w) or 10.0% (w / w). In addition, in such an embodiment, preferably dicaprylyl carbonate and ethylhexyl cocoate are used in equal amounts, for example, 2.0% (w / w) dicaprylyl carbonate is combined with 2.0% (w / w) ethylhexyl cocoate, or 5.0% (w / w) dicaprylyl carbonate is combined with 5.0% (w / w) ethylhexyl cocoate.

[0052] Thickener

[0053] Preferably, the skin care composition further comprises a thickening agent. A thickening agent is a compound that increases the viscosity of a cosmetic or pharmaceutical formulation. A thickening agent is usually a polymer that absorbs water and swells, thereby making the composition more viscous. Thickening agents commonly used in skin care products include guar gum, xanthan gum, gelatin, carnauba wax and stearic acid.

[0054] According to a preferred embodiment of the present invention, the skin care composition comprising a freeze-dried or spray-dried live bacterium of at least one Propionibacterium acnes strain and an emollient as defined above comprises a thickening agent selected from carrageenan extract, hydroxypropyl starch phosphate and mixtures thereof. In a preferred embodiment, the skin care composition comprises a freeze-dried or spray-dried live bacterium of at least one Propionibacterium acnes strain, an emollient as defined above, and carrageenan extract as a thickening agent. In another preferred embodiment, the skin care composition comprises a freeze-dried or spray-dried live bacterium of at least one Propionibacterium acnes strain, an emollient as defined above, and hydroxypropyl starch phosphate as a thickening agent.

[0055] When using carrageenan extract as a thickening compound in the composition of the present invention, it is preferably used in the final skin care composition in an amount of 0.05 to 7.5% (w / w), more preferably 0.1 to 5.0% (w / w), more preferably 0.2 to 4.0% (w / w), 0.2 to 2.0% (w / w) or 0.2 to 1.5% (w / w). In other words, the amount of carrageenan extract in the skin care composition of the present invention can be at least 0.05% (w / w), at least 0.1% (w / w), at least 0.25% (w / w), at least 0.5% (w / w), at least 0.75% (w / w), at least 1.0% (w / w), at least 1.25% (w / w), at least 1.5% (w / w), at least 1.75% (w / w), at least 2.0% (w / w), at least 2.5% (w / w), at least 3.0% (w / w), at least 4.0% (w / w) or at least 5.0% (w / w).

[0056] When using hydroxypropyl starch phosphate as a thickening agent, it is preferably used in the final skin care composition in an amount of 0.05 to 10.0% (w / w), more preferably 0.1 to 10.0% (w / w), more preferably 0.5 to 7.5% (w / w), 1.0 to 5.0% (w / w) or 1.0 to 2.0% (w / w). In other words, the amount of hydroxypropyl starch phosphate in the skin care composition of the present invention can be at least 0.05% (w / w), at least 0.1% (w / w), at least 0.25% (w / w), at least 0.5% (w / w), at least 0.75% (w / w), at least 1.0% (w / w), at least 1.25% (w / w), at least 1.5% (w / w) or at least 1.75% (w / w).

[0057] It should be understood that the skin care composition of the present invention may also contain a combination of carrageenan extract and hydroxypropyl starch phosphate as a thickener. When the carrageenan extract and hydroxypropyl starch phosphate are used in combination with each other as a thickener, preferably the total amount of the thickener is at least 0.05% (w / w), but not more than 10.0% (w / w), more preferably not more than 5.0% (w / w). In addition, in such an embodiment, preferably the two thickeners are used in equal amounts, for example, 2.0% (w / w) carrageenan extract and 2.0% (w / w) hydroxypropyl starch phosphate.

[0058] pH regulator

[0059] In another preferred case, the skin care composition of the present invention further contains a pH regulator. Since the composition of the present invention is used on human skin, it preferably has a neutral or slightly acidic pH to be compatible with the acidic environment of the skin. The composition may have a pH in the range of about 2.5 to about 7.5, preferably about 4.0 to about 7.0, more preferably about 6.0 to about 7.0. Although an acidic pH in a cosmetic preparation is usually achieved by adding acids such as formic acid, acetic acid, butyric acid, valeric acid, hexanoic acid, heptanoic acid or octanoic acid, it has been found that these acids may impair the ability of the composition to support bacterial growth and replication on the skin after administration. According to the present invention, the pH regulator is preferably a citric acid / citrate buffer.

[0060] Therefore, in another preferred case, the skin care composition comprising the freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain and the emollient as defined above further contains a citric acid / citrate buffer such as a citric acid / sodium citrate buffer as a pH regulator. In a preferred embodiment, the skin care composition comprises the freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, the emollient as defined above, and a buffer composed of citric acid and sodium citrate. Particularly preferably, the skin care composition does not contain any other pH regulator except citric acid / citrate.

[0061] When citric acid is used as a pH regulator, it is preferably used in the final skin care composition in an amount of 0.01 to 1.5% (w / w), more preferably 0.01 to 0.25% (w / w), even more preferably 0.01 to 0.1% (w / w). In other words, the amount of citric acid in the skin care composition of the present invention can be at least 0.01% (w / w), at least 0.02% (w / w), at least 0.03% (w / w), at least 0.04% (w / w), at least 0.05% (w / w), at least 0.06% (w / w), at least 0.07% (w / w), at least 0.08% (w / w), at least 0.09% (w / w) or at least 0.1% (w / w).

[0062] When sodium citrate is used as a pH regulator, it is preferably used in the final skin care composition in an amount of 0.01 to 1.5% (w / w), more preferably 0.05 to 1.0% (w / w), more preferably 0.05 to 0.5% (w / w) or 0.1 to 0.2% (w / w). In other words, the amount of sodium citrate in the skin care composition of the present invention can be at least 0.01% (w / w), at least 0.02% (w / w), at least 0.03% (w / w), at least 0.04% (w / w), at least 0.05% (w / w), at least 0.06% (w / w), at least 0.07% (w / w), at least 0.08% (w / w), at least 0.09% (w / w), at least 0.1% (w / w), 0.11% (w / w), 0.12% (w / w), 0.13% (w / w), 0.14% (w / w), 0.15% (w / w), 0.16% (w / w), 0.17% (w / w), 0.18% (w / w), 0.19% (w / w), 0.2% (w / w), at least 0.3% (w / w), at least 0.4% (w / w), at least 0.5% (w / w), at least 0.6% (w / w), at least 0.7% (w / w), at least 0.8% (w / w), at least 0.9% (w / w) or at least 1.0%.

[0063] When citric acid and sodium citrate are used in combination with each other as a pH regulator, preferably the total amount of the pH regulator in the final skin care composition does not exceed 0.3% (w / w). In addition, in such an embodiment, preferably citric acid and sodium citrate are used in a ratio of 1:2, for example 0.05% (w / w) citric acid and 0.10% (w / w) sodium citrate, or 0.1% (w / w) citric acid combined with 0.2% (w / w) sodium citrate.

[0064] Particularly preferably, citric acid and citrate are used in the skin care composition of the present invention in an amount that ensures that the pH of the total composition is between 5.0 and 7.0.

[0065] Filler

[0066] In yet another preferred case, the skin care composition further comprises a filler. As used herein, a filler is a compound that helps to make the composition more homogeneous by being uniformly dispersed in the skin care composition. Fillers are used to improve the sensory properties of the skin. Depending on the filler material, the final product may impart a silky, dry, smooth or powdery skin feel.

[0067] According to another preferred case of the present invention, the skin care composition comprising the freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain and the emollient defined above comprises a filler selected from distarch phosphate, tapioca starch and mixtures thereof. In a preferred embodiment, the skin care composition comprises the freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, the emollient as defined above, and distarch phosphate as a filler. In another preferred embodiment, the skin care composition comprises the freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, the emollient as defined above, and tapioca starch as a filler. In yet another preferred embodiment, the skin care composition comprises the freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, the emollient as defined above, and both distarch phosphate and tapioca starch as fillers. Preferably, the skin care composition does not contain any other filler other than distarch phosphate and / or tapioca starch.

[0068] When using distarch phosphate as a filler, it is preferably used in an amount of 0.05 to 5.0% (w / w), more preferably 1.0 to 5.0% (w / w), more preferably 2.0 to 5.0% (w / w) or 2.0 to 3.0% (w / w) in the final skin care composition. In other words, the amount of distarch phosphate in the skin care composition of the present invention can be at least 0.05% (w / w), at least 0.1% (w / w), at least 0.25% (w / w), at least 0.5% (w / w), at least 0.75% (w / w), at least 1.0% (w / w), at least 1.25% (w / w), at least 1.5% (w / w), at least 1.75% (w / w), at least 2.0% (w / w), at least 2.5% (w / w), at least 3.0% (w / w) or at least 4.0% (w / w). The same amounts apply when using tapioca starch as a filler.

[0069] When distarch phosphate and tapioca starch are used in combination with each other as fillers, preferably the total amount of the fillers is at least 0.05% (w / w), but not more than 5.0% (w / w), more preferably not more than 3.0% (w / w). In addition, in such an embodiment, preferably distarch phosphate and tapioca starch are used in equal amounts, for example, 2.5% (w / w) distarch phosphate is combined with 2.5% (w / w) tapioca starch, or 1.0% (w / w) distarch phosphate is combined with 1.0% (w / w) tapioca starch.

[0070] Solubilizer

[0071] In another preferred case, the skin care composition further comprises a solubilizer. As used herein, a solubilizer is a compound that aids in the dissolution of hydrophobic substances in aqueous and alcoholic formulations. For example, a solubilizer can make the dissolution of perfume oils and other hydrophobic substances such as vitamins in an aqueous skin care composition feasible. It has been found herein that polyethylene glycol (PEG)-40 hydrogenated castor oil does not interfere with the viability and replication ability of the freeze-dried or spray-dried bacteria.

[0072] Thus, in a preferred embodiment, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, an emollient as defined above, and PEG-40 hydrogenated castor oil. In an even more preferred embodiment, PEG-40 hydrogenated castor oil is the only solubilizer comprised in the composition.

[0073] When PEG-40 hydrogenated castor oil is used as a solubilizer, it is preferably used in an amount of 0.01 to 2.5% (w / w), more preferably 0.05 to 1.5% (w / w), even more preferably 0.5 to 1.0% (w / w) in the final skin care composition. In other words, the amount of PEG-40 hydrogenated castor oil in the skin care composition of the present invention can be at least 0.01% (w / w), at least 0.05% (w / w), at least 0.1% (w / w), at least 0.2% (w / w), at least 0.3% (w / w), at least 0.4% (w / w), at least 0.5% (w / w), at least 0.6% (w / w), at least 0.7% (w / w), at least 0.8% (w / w), at least 0.9% (w / w) or at least 1.0% (w / w).

[0074] Antioxidant

[0075] In another preferred case, the skin care composition further comprises an antioxidant. These compounds are typically added to cosmetic formulations to prevent oxidation reactions catalyzed by oxygen free radicals, which would otherwise cause the decomposition of ingredients in the composition such as proteins, sugars, and lipids. Antioxidants commonly used in beauty products include chemicals such as butylated hydroxytoluene and butylated hydroxyanisole, as well as polyphenols, flavonoids, flavanols, stilbenes, and terpenes of plant origin.

[0076] In a preferred embodiment, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, an emollient as defined above, and an antioxidant selected from tocopherol, tocopheryl acetate, and mixtures thereof. In a preferred embodiment, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, an emollient as defined above, and tocopherol as an antioxidant. In another preferred embodiment, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, an emollient as defined above, and tocopheryl acetate as an antioxidant. In yet another preferred embodiment, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, an emollient as defined above, and both tocopherol and tocopheryl acetate as antioxidants. Particularly preferably, the skin care composition does not contain any other antioxidant other than tocopherol and / or tocopheryl acetate.

[0077] When tocopherol, tocopheryl acetate, or a mixture thereof is used as an antioxidant, they can be added in freeze-dried or spray-dried form to the freeze-dried or spray-dried live bacteria. For example, if the composition of the present invention is provided in a form that requires mixing the bacteria with a cosmetic or pharmaceutical formulation before use, the freeze-dried or spray-dried antioxidant can be added to the freeze-dried or spray-dried bacteria and stored until reconstituted with the cosmetic or pharmaceutical formulation before use. When tocopherol, tocopheryl acetate, or a mixture thereof is used in dry form, dry vitamin C can also be added.

[0078] When using tocopherol as an antioxidant, it is preferably used in the final skin care composition in an amount of 0.01 to 2.0% (w / w), more preferably 0.05 to 1.5% (w / w), even more preferably 0.1 to 1.0% (w / w). In other words, the amount of tocopherol in the skin care composition of the present invention can be at least 0.01% (w / w), at least 0.025% (w / w), at least 0.05% (w / w), at least 0.075% (w / w), at least 0.1% (w / w), at least 0.2% (w / w), at least 0.3% (w / w), at least 0.4% (w / w), at least 0.5% (w / w), at least 0.6% (w / w), at least 0.7% (w / w), at least 0.8% (w / w), at least 0.9% (w / w) or at least 1.0% (w / w). The same amounts apply when using tocopheryl acetate as an antioxidant.

[0079] When using tocopherol and tocopheryl acetate in combination with each other as antioxidants, preferably the total amount of the antioxidants is at least 0.01% (w / w), but does not exceed 2.0% (w / w). In addition, in such an embodiment, preferably tocopherol and tocopheryl acetate are used in equal amounts, for example, 0.25% (w / w) tocopherol is combined with 0.25% (w / w) tocopheryl acetate, or 0.5% (w / w) tocopherol is combined with 0.5% (w / w) tocopheryl acetate.

[0080] Preservative

[0081] In another preferred case, the skin care composition further comprises a preservative. As used herein, a preservative is a compound added to a beauty preparation to prevent microbial spoilage of the preparation by inhibiting the growth of accidental bacteria and yeasts. Common preservatives in beauty preparations include benzyl alcohol, salicylic acid, and sorbic acid, etc.

[0082] In a preferred embodiment, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, an emollient as defined above, and a preservative selected from ethanol, phenoxyethanol, octylene glycol, methylpropanediol, and mixtures thereof. These preservatives are surprisingly tolerated to a certain extent by the Propionibacterium acnes strain while effectively combating other bacteria that may contaminate the composition.

[0083] In a preferred embodiment, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, an emollient as defined above, and ethanol as a preservative. In another preferred embodiment, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, an emollient as defined above, and phenoxyethanol as a preservative. In yet another preferred embodiment, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, an emollient as defined above, and caprylyl glycol as a preservative. In yet another preferred embodiment, the skin care composition comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, an emollient as defined above, and methylpropanediol as a preservative.

[0084] In addition, the skin care composition of the present invention may comprise more than one preservative, such as two or more of the above-mentioned preservatives. For example, the composition may comprise a combination of ethanol and phenoxyethanol as a preservative. Alternatively, the composition may comprise a combination of ethanol and caprylyl glycol as a preservative. Alternatively, the composition may comprise a combination of ethanol and methylpropanediol as a preservative. The composition may also comprise a combination of phenoxyethanol and caprylyl glycol as a preservative or a combination of phenoxyethanol and methylpropanediol as a preservative. The composition may also comprise a combination of caprylyl glycol and methylpropanediol as a preservative.

[0085] The best results have been obtained using a combination of phenoxyethanol, caprylyl glycol, and methylpropanediol in protecting the skin care composition of the present invention from spoilage by other bacteria or fungi while maintaining the viability of the freeze-dried or spray-dried live Propionibacterium acnes bacteria in the composition. Thus, in a particular preferred embodiment, the skin care composition of the present invention comprises freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, an emollient, and a combination of phenoxyethanol, caprylyl glycol, and methylpropanediol.

[0086] When ethanol is used as a preservative, it is preferably used in the final skin care composition in an amount of 0.5 to 20.0% (w / w), more preferably 8.0 to 15.0% (w / w), even more preferably 5.0 to 15.0% (w / w). In other words, the amount of ethanol in the skin care composition of the present invention can be at least 0.5% (w / w), at least 1.0% (w / w), at least 2.0% (w / w), at least 3.0% (w / w), at least 4.0% (w / w), at least 5.0% (w / w), at least 6.0% (w / w), at least 7.0% (w / w), at least 8.0% (w / w), at least 9.0% (w / w), or at least 10.0% (w / w).

[0087] When phenoxyethanol is used as a preservative, it is preferably used in the final skin care composition in an amount of 0.05 to 0.5% (w / w), more preferably 0.1 to 0.25% (w / w), still more preferably 0.1 to 0.2% (w / w). In other words, the amount of phenoxyethanol in the skin care composition of the present invention can be at least 0.05% (w / w), at least 0.075% (w / w), at least 0.1% (w / w), at least 0.15% (w / w), at least 0.2% (w / w) or at least 0.25% (w / w). The same amounts apply when caprylyl glycol is used as a preservative.

[0088] When methylpropanediol is used as a preservative, it is preferably used in the final skin care composition in an amount of 0.05 to 5.0% (w / w), more preferably 0.5 to 2.5% (w / w), still more preferably 1.0 to 1.5% (w / w). In other words, the amount of methylpropanediol in the skin care composition of the present invention can be at least 0.5% (w / w), at least 1.0% (w / w), at least 2.0% (w / w), at least 3.0% (w / w), at least 4.0% (w / w) or at least 5.0% (w / w).

[0089] When two or more of the above-mentioned preservatives are used in combination with each other, preferably the total amount of the preservatives does not exceed 5.0% (w / w) when using a combination of phenoxyethanol and caprylyl glycol. Specifically, when using a combination of caprylyl glycol, phenoxyethanol and methylpropanediol, these compounds are preferably used in a ratio of 1:2:20, such as 0.05% (w / w) caprylyl glycol, 0.1% (w / w) phenoxyethanol and 1.0% (w / w) methylpropanediol, or 0.1% (w / w) caprylyl glycol, 0.2% (w / w) phenoxyethanol and 2.0% (w / w) methylpropanediol.

[0090] The above-mentioned preservatives can be used in combination with other preservatives, especially short-chain fatty acids such as formic acid, propionic acid or isobutyric acid. In addition, the composition of the present invention may also contain lactic acid as an additional preservative.

[0091] Other cosmetic excipients

[0092] The skin care composition described herein may further include known excipients in addition to the above components, including perfumes, pigments, colorants, dyes, waxes, masking agents, humectants, surfactants, lubricants, stabilizers, sunscreens, emulsifiers, drugs, antibacterial agents, chelating agents, protectants, thickeners, vitamins, panthenol, ubiquinone Q10, hyaluronic acid or any combination thereof.

[0093] Bacterial strain

[0094] The skin care composition of the present invention comprises freeze-dried or spray-dried live bacteria of at least one strain of the species Cutibacterium acnes. However, preferably the skin care composition comprises two or more strains of the species Cutibacterium acnes. For example, the skin care composition may comprise 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more than 20 strains of Cutibacterium acnes. In certain embodiments, the skin care composition comprises 2, 3, 4 or 5 different strains of Cutibacterium acnes. It has been reported that the reactivation and growth of certain Cutibacterium acnes strains are significantly supported when grown in a mixture of strains compared to when grown alone. For example, the SLST K8 type strain grows slowly when grown alone but significantly faster when grown in a mixture of strains. Thus, according to the present invention, the composition preferably comprises two or more strains of Cutibacterium acnes.

[0095] The bacterial species Cutibacterium acnes (C. acnes) was formerly known as Propionibacterium acnes (P. acnes). Based on biochemical and genomic studies, the species was taxonomically reclassified in 2016. Cutibacterium acnes is a Gram-positive anaerobic rod-shaped bacterium known to be involved in the development of acne and other pathological conditions. Cutibacterium acnes strains are present on the skin of most people. Cutibacterium acnes strains can be pathogenic or non-pathogenic. As used herein, a "pathogenic" Cutibacterium acnes strain is a strain associated with acne. Assays for the identification and selection of pathogenic and non-pathogenic Cutibacterium acnes strains are described in WO 2018 / 073651.

[0096] It has been shown that Cutibacterium acnes comprises several distinct major phylogenetic groups classified as types I, II and III, where the major type I clade is further divided into sub-clades called types IA, IB and IC (Lomholt and Kilian, 2010). Sub-clade IA is further subdivided into IA1 and IA2 (McDowell et al., 2012). Preferably, at least one strain of the species Cutibacterium acnes is a non-pathogenic strain of Cutibacterium acnes. Genetic analysis of Cutibacterium acnes strains has shown that non-pathogenic strains not associated with acne are mainly members of (i) sub-clade IA2 of clade I, (ii) sub-clade IB of clade I and (iii) clade II.

[0097] Thus, in one embodiment of the present invention, at least one strain of the species Propionibacterium acnes belongs to any one of the subclades IA2 and IB of clade I or belongs to clade II. In one embodiment, at least one strain of the species Propionibacterium acnes belongs to subclade IA2. In another embodiment, at least one strain of the species Propionibacterium acnes belongs to subclade IB. In yet another embodiment, at least one strain of the species Propionibacterium acnes belongs to clade II. If more than one strain is used in the skin care composition of the present invention, preferably strains from different clades or subclades are mixed with each other. For example, in one embodiment, the skin care composition comprises at least one strain from subclade IA2 and at least one strain from subclade IB. In another embodiment, the skin care composition comprises at least one strain from subclade IA2 and at least one strain from clade II. In yet another embodiment, the skin care composition comprises at least one strain from subclade IB and at least one strain from clade II.

[0098] In other embodiments, the skin care composition of the present invention may comprise a mixture of Propionibacterium acnes strains, which includes one or more clade I strains and one or more clade II strains. Although, as noted above, the pathogenicity of clade II strains may be lower than that of clade I strains, these strains may also grow more slowly than clade I strains and are less likely to be able to colonize the skin on their own. Thus, in certain embodiments, it may be advantageous for the skin care composition to comprise a strain mixture that includes both clade I and clade II strains, which allows for relatively improved colonization of the skin by clade II strains.

[0099] Non-limiting examples of non-pathogenic strains of Propionibacterium acnes include, but are not limited to, SLST D1, A5, C1, C3, H1, H2, H3, K1, K2, K4, K6, K8, K9, L1, and F4 type strains. Particularly preferably, the skin care composition of the present invention comprises at least one SLST C3 type strain and / or at least one SLST K8 type strain. In one embodiment, the skin care composition of the present invention comprises at least one SLST C3 type strain, more preferably two or more SLST C3 type strains, such as 2, 3, 4, 5, 6, 7, 8, 9, or 10 SLST C3 type strains. In another embodiment, the skin care composition of the present invention comprises at least one SLST K8 type strain, more preferably two or more SLST K8 type strains, such as 2, 3, 4, 5, 6, 7, 8, 9, or 10 SLST K8 type strains. In yet another embodiment, the skin care composition of the present invention comprises a combination of at least one SLST C3 type strain and at least one SLST K8 type strain. In yet another embodiment, the skin care composition of the present invention comprises a combination of two or more SLST C3 type strains and at least one SLST K8 type strain. In yet another embodiment, the skin care composition of the present invention comprises a combination of at least one SLST C3 type strain and two or more SLST K8 type strains. In yet another embodiment, the skin care composition of the present invention comprises a combination of two or more SLST C3 type strains and two or more SLST K8 type strains. In certain embodiments, the one or more SLST C3 type strains and the one or more SLST K8 type strains in the composition have approximately equal concentrations. In other embodiments, the one or more SLST C3 type strains in the composition have a higher concentration than the one or more SLST K8 type strains. In other embodiments, the one or more SLST C3 type strains in the composition have a lower concentration than the one or more SLST K8 type strains.

[0100] In a particularly preferred embodiment, the skin care composition of the present invention comprises the SLST C3 strain with accession number DSM 32667 deposited on October 19, 2017 at the Leibniz Institute DSMZ - German Collection of Microorganisms and Cell Cultures (Inhoffenstr. 7B, D-38124 Braunschweig, Germany) by S-Biomedic N.V. (Turnhoutsweg 30, 2340 Beerse, Belgium) under the Budapest Treaty. In another particularly preferred embodiment, the skin care composition of the present invention comprises the SLST K8 strain with accession number DSM 32668 deposited on October 19, 2017 at the Leibniz Institute DSMZ - German Collection of Microorganisms and Cell Cultures (Inhoffenstr. 7B, D-38124 Braunschweig, Germany) by S-Biomedic N.V. (Turnhoutsweg 30, 2340 Beerse, Belgium) under the Budapest Treaty. In yet another preferred embodiment, the skin care composition of the present invention comprises both the SLST C3 strain DSM 32667 and the SLST K8 strain DSM 32668. In another embodiment, the skin care composition comprising at least one SLST C3 strain and at least one SLST K8 strain further comprises at least one SLST A5 strain and / or at least one SLST F4 strain. Thus, in one embodiment, the composition comprises at least one SLST C3 strain, at least one SLST K8 strain, and at least one SLST A5 strain. In another embodiment, the composition comprises at least one SLST C3 strain, at least one SLST K8 strain, and at least one SLST F4 strain. In yet another embodiment, the composition comprises at least one SLST C3 strain, at least one SLST K8 strain, at least one SLST A5 strain, and at least one SLST F4 strain.

[0101] The strain names referred to in this text are based on the single-locus sequence typing (SLST) scheme using the locus PPA2385 as the SLST target sequence as described in Scholz et al., 2014. The sequences of the PPA2385 locus of the different strains identified by Scholz are listed herein as SEQ ID NOs: 1-76. Thus, a "SLST C3 type" strain is a strain that contains in its genome a PPA2385 locus sequence that is 100% identical to the sequence depicted in SEQ ID NO: 27. Similarly, a "SLST K8 type" strain is a strain that contains in its genome a PPA2385 locus sequence that is 100% identical to the sequence depicted in SEQ ID NO: 64. The SLST scheme is also described in more detail in WO 2018 / 073651. The sequence identification of the PPA2385 locus can be carried out by PCR amplification and DNA sequencing using the nucleotide primer sets set forth in SEQ ID NOs: 77-82 as described in WO 2018 / 073651. Based on the information presented herein and in WO 2018 / 073651, one of ordinary skill in the art would understand how to identify and classify Propionibacterium acnes strains.

[0102] The compositions of the present invention may include both pathogenic and non-pathogenic strains of Propionibacterium acnes. However, in a preferred embodiment of the present invention, the skin care composition contains only non-pathogenic strains of Propionibacterium acnes. Particularly preferably, the skin care composition of the present invention does not include the ribotype 6 (RT6) strain of Propionibacterium acnes. The ribotype classification system is based on differences in the 16S rDNA sequences between different strains of Propionibacterium acnes. The ribotype system is explained in, for example, Fitz-Gibbon et al., 2013. Additionally particularly preferably, the skin care composition of the present invention does not include the phenotypic type III strain of Propionibacterium acnes.

[0103] Strains of Propionibacterium acnes are generally able to produce the signaling molecule trans-10,cis-12 linoleic acid from the precursor molecule linoleic acid that is naturally present in sebum (Rosson et al., 2004). trans-10,cis-12 Linoleic acid is thought to stimulate sebum production and secretion, which is important for the skin colonization of Propionibacterium acnes. In this way, trans-10,cis-12 linoleic acid promotes the onset of acne (Downing et al., 1986; Letawe et al., 1998). Depending on the skin of the subject, reducing or increasing sebum production may be beneficial. For example, reducing sebum production in the skin of a subject with acne or oily skin may be useful. Conversely, increasing sebum production in the skin of a subject with dry skin may be useful.

[0104] Thus, in one embodiment, one or more Propionibacterium acnes strains included in the skin care composition of the present invention are selected based on their ability to produce trans-10, cis-12 linoleic acid. In a preferred embodiment, strains that produce low levels of trans-10, cis-12 linoleic acid are selected and included in the skin care composition of the present invention for combating acne or oily skin. Without wishing to be bound by theory, it is believed that these strains reduce sebum production, which is useful for preventing or alleviating the symptoms of acne or oily skin. SLST C3, C1, F4, A5, K1, K2, K8, and L1 type strains produce only small amounts of trans-10, cis-12 linoleic acid. In another preferred embodiment, strains that produce high levels of trans-10, cis-12 linoleic acid are selected and included in the skin care composition of the present invention for combating dry skin. Without wishing to be bound by theory, it is believed that these strains increase sebum production, which is useful for preventing or alleviating the symptoms of dry skin. SLST A1 type strain produces large amounts of trans-10, cis-12 linoleic acid.

[0105] In one embodiment, one or more Propionibacterium acnes strains included in the skin care composition of the present invention are isolated from the skin microbiome of a donor subject. The subject may not have acne or oily skin, or may have mild, moderate, or severe acne. In another embodiment, the strains isolated from the skin microbiome of the donor subject are non-pathogenic strains.

[0106] The skin care composition of the present invention may also contain one or more genetically modified Propionibacterium acnes strains. In another embodiment, the skin care composition contains a combination of one or more genetically modified Propionibacterium acnes strains and one or more naturally occurring Propionibacterium acnes strains. The genetically modified strains are preferably modified to produce lower or higher amounts of trans-10, cis-12 linoleic acid. The production of trans-10, cis-12 linoleic acid can be detected as described in U.S. Patent 6,743,609 or by other well-known methods such as FAME (fatty acid methyl ester) or gas chromatography. In some other embodiments, the skin care composition of the present invention contains only naturally occurring Propionibacterium acnes strains, i.e., only Propionibacterium acnes strains that have not been genetically modified by humans.

[0107] Particularly preferably, the at least one Propionibacterium acnes strain is at 1.0 x 10 4 -1.0 x 10 11 colony forming units (CFU) / ml, more preferably 1.0 x 10 5 -1.0 x 10 10 CFU / ml, even more preferably 1.0 x 10 7 -1.0 x10 10CFU / ml or 1.0 x 10 8 -1.0 x 10 9 CFU / ml is present in the composition. For example, the at least one Propionibacterium acnes strain may be present in an amount of at least 1.0 x 10 5 CFU / ml, preferably at least 1.0 x 10 6 CFU / ml, more preferably at least 1.0 x 10 7 CFU / ml, for example at least 1.0 x 10 8 CFU / ml, at least 1.0 x 10 9 CFU / ml or at least 1.0 x10 10 CFU / ml is present in the skin care composition. Particularly preferably, the at least one Propionibacterium acnes strain is present in an amount of at least 1.0 x 10 10 CFU / ml, 2.0 x 10 10 CFU / ml, 3.0 x 10 10 CFU / ml, 4.0 x 10 10 CFU / ml, 5.0 x 10 10 CFU / ml, 6.0 x 10 10 CFU / ml, 7.0 x 10 10 CFU / ml, 8.0 x 10 10 CFU / ml or 9.0 x10 10 CFU / ml is present in the skin care composition.

[0108] In certain embodiments, each Propionibacterium acnes strain present in the composition is present in an amount of 1.0 x 10 4 -1.0 x 10 11 CFU / ml, more preferably 1.0 x 10 5 -1.0 x 10 10 CFU / ml, even more preferably 1.0 x 10 7 -1.0x 10 10 CFU / ml or 1.0 x 10 8 -1.0 x 10 9 CFU / ml. For example, each Propionibacterium acnes strain may be present in an amount of at least 1.0 x 10 5 CFU / ml, preferably at least 1.0 x 10 6 CFU / ml, more preferably at least 1.0 x 10 7 CFU / ml, for example at least 1.0 x 108 CFU / ml, at least 1.0 x 10 9 CFU / ml or at least 1.0 x 10 10 CFU / ml is present in the skin care composition. Particularly preferably, each Propionibacterium acnes strain is present in an amount of at least 1.0 x 10 10 CFU / ml, 2.0 x 10 10 CFU / ml, 3.0 x 10 10 CFU / ml, 4.0 x 10 10 CFU / ml, 5.0 x 10 10 CFU / ml, 6.0 x10 10 CFU / ml, 7.0 x 10 10 CFU / ml, 8.0 x 10 10 CFU / ml or 9.0 x 10 10 CFU / ml is present in the skin care composition. For example, if the skin care composition of the present invention contains an SLST C3 strain and an SLST K8 strain, each of these strains can be present in an amount of 1.0 x 10 4 -1.0 x 10 11 CFU / ml, for example 1.0 x10 10 CFU / ml, 2.0 x 10 10 CFU / ml, 3.0 x 10 10 CFU / ml, 4.0 x 10 10 CFU / ml, 5.0 x 10 10 CFU / ml, 6.0 x 10 10 CFU / ml, 7.0 x 10 10 CFU / ml, 8.0 x 10 10 CFU / ml or 9.0 x 10 10 CFU / ml is present.

[0109] In certain embodiments, the total amount of freeze-dried or spray-dried bacteria in the composition is 1.0 x10 4 -1.0 x 10 11 CFU / ml, more preferably 1.0 x 10 5 -1.0 x 10 10 CFU / ml, even more preferably 1.0 x10 7 -1.0 x 10 10 CFU / ml or 1.0 x 10 8-1.0 x 10 9 CFU / ml. For example, the bacteria can be combined to be at least 1.0 x 10 5 CFU / ml, preferably at least 1.0 x 10 6 CFU / ml, more preferably at least 1.0 x 10 7 CFU / ml. For example, at least 1.0 x 10 8 CFU / ml, at least 1.0 x 10 9 CFU / ml or at least 1.0 x 10 10 CFU / ml of the skin care composition is present in the composition. Particularly preferably, the bacteria are combined to be at least 1.0 x 10 10 CFU / ml, 2.0 x10 10 CFU / ml, 3.0 x 10 10 CFU / ml, 4.0 x 10 10 CFU / ml, 5.0 x 10 10 CFU / ml, 6.0 x 10 10 CFU / ml, 7.0 x 10 10 CFU / ml, 8.0 x 10 10 CFU / ml or 9.0 x 10 10 CFU / ml of the skin care composition is present in the composition. Those of ordinary skill in the art can easily determine the amount of bacteria in the freeze-dried or spray-dried composition.

[0110] Preferred composition

[0111] The above components described as being suitable for the skin care composition of the present invention can be combined with each other without limitation. Therefore, in addition to the freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain and the emollient, the skin care composition of the present invention may further comprise at least one excipient selected from thickeners, fillers, antioxidants, preservatives, solubilizers and pH regulators, wherein the one or more excipients do not interfere with the viability of the freeze-dried or spray-dried live bacteria and the reactivation of growth. Particularly preferred skin care compositions covered by the present disclosure are described below.

[0112] A preferred skin care composition comprises

[0113] (a) freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain; and

[0114] (b) Dioctyl carbonate from 0.05 to 25.0% (w / w), preferably from 7.5 to 10.0% (w / w) of dioctyl carbonate.

[0115] Another preferred skin care composition comprises

[0116] (a) Freeze-dried or spray-dried live bacteria of at least one strain of Propionibacterium acnes; and

[0117] (b) Ethylhexyl cocoate from 0.05 to 25.0% (w / w), preferably from 7.5 to 10.0% (w / w) of ethylhexyl cocoate.

[0118] A preferred skin care composition comprises

[0119] (a) Freeze-dried or spray-dried live bacteria of at least one strain of Propionibacterium acnes selected from the SLST D1, A5, C1, C3, H1, H2, H3, K1, K2, K4, K6, K8, K9, L1 and F4 types; and

[0120] (b) Dioctyl carbonate from 0.05 to 25.0% (w / w), preferably from 7.5 to 10.0% (w / w) of dioctyl carbonate.

[0121] Another preferred skin care composition comprises

[0122] (a) Freeze-dried or spray-dried live bacteria of at least one strain of Propionibacterium acnes selected from the SLST D1, A5, C1, C3, H1, H2, H3, K1, K2, K4, K6, K8, K9, L1 and F4 types; and

[0123] (b) Ethylhexyl cocoate from 0.05 to 25.0% (w / w), preferably from 7.5 to 10.0% (w / w) of ethylhexyl cocoate.

[0124] A preferred skin care composition comprises

[0125] (a) Freeze-dried or spray-dried live bacteria of at least one strain of Propionibacterium acnes SLST C3 type and at least one strain of Propionibacterium acnes SLST K8 type; and

[0126] (b) Dioctyl carbonate from 0.05 to 25.0% (w / w), preferably from 7.5 to 10.0% (w / w) of dioctyl carbonate.

[0127] Another preferred skin care composition comprises

[0128] (a)At least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes SLST C3 type strain and at least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes SLST K8 type strain; and

[0129] (b)0.05 to 25.0% (w / w) of ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) of ethylhexyl cocoate.

[0130] Another preferred skin care composition comprises

[0131] (a)At least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes strain, preferably at least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes SLST C3 type strain and at least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes SLST K8 type strain;

[0132] (b)0.05 to 25.0% (w / w) of dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) of dicaprylyl carbonate; and

[0133] (c)0.05 to 7.5% (w / w) of Chondrus crispus extract, preferably 0.2 or 1.5% (w / w) of Chondrus crispus extract.

[0134] Another preferred skin care composition comprises

[0135] (a)At least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes strain, preferably at least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes SLST C3 type strain and at least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes SLST K8 type strain;

[0136] (b)0.05 to 25.0% (w / w) of ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) of ethylhexyl cocoate; and

[0137] (c)0.05 to 7.5% (w / w) of Chondrus crispus extract, preferably 0.2 or 1.5% (w / w) of Chondrus crispus extract.

[0138] Another preferred skin care composition comprises

[0139] (a)At least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes strain, preferably at least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes SLST C3 type strain and at least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes SLST K8 type strain;

[0140] (b)0.05 to 25.0% (w / w) of dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) of dicaprylyl carbonate; and

[0141] (c) 0.05 to 10.0% (w / w) of hydroxypropyl starch phosphate, preferably 2.0 to 4.0% (w / w) of hydroxypropyl starch phosphate.

[0142] Another preferred skin care composition comprises

[0143] (a) lyophilized or spray-dried live bacteria of at least one strain of Propionibacterium acnes, preferably lyophilized or spray-dried live bacteria of at least one strain of Propionibacterium acnes SLST C3 type and at least one strain of Propionibacterium acnes SLST K8 type;

[0144] (b) 0.05 to 25.0% (w / w) of ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) of ethylhexyl cocoate; and

[0145] (c) 0.05 to 10.0% (w / w) of hydroxypropyl starch phosphate, preferably 2.0 to 4.0% (w / w) of hydroxypropyl starch phosphate.

[0146] Another preferred skin care composition comprises

[0147] (a) lyophilized or spray-dried live bacteria of at least one strain of Propionibacterium acnes, preferably lyophilized or spray-dried live bacteria of at least one strain of Propionibacterium acnes SLST C3 type and at least one strain of Propionibacterium acnes SLST K8 type;

[0148] (b) 0.05 to 25.0% (w / w) of dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) of dicaprylyl carbonate; and

[0149] (c) a citric acid / citrate buffer that provides the composition with a pH of about 4.0 to about 7.0.

[0150] Another preferred skin care composition comprises

[0151] (a) lyophilized or spray-dried live bacteria of at least one strain of Propionibacterium acnes, preferably lyophilized or spray-dried live bacteria of at least one strain of Propionibacterium acnes SLST C3 type and at least one strain of Propionibacterium acnes SLST K8 type;

[0152] (b) 0.05 to 25.0% (w / w) of ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) of ethylhexyl cocoate; and

[0153] (c) a citric acid / citrate buffer that provides the composition with a pH of about 4.0 to about 7.0.

[0154] Another preferred skin care composition comprises

[0155] (a) freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0156] (b) 0.05 to 25.0% (w / w) of dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) of dicaprylyl carbonate; and

[0157] (c) 0.05 to 5.0% (w / w) of distarch phosphate, preferably 2.0 to 3.0% (w / w) of distarch phosphate.

[0158] Another preferred skin care composition comprises

[0159] (a) freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0160] (b) 0.05 to 25.0% (w / w) of ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) of ethylhexyl cocoate; and

[0161] (c) 0.05 to 5.0% (w / w) of distarch phosphate, preferably 2.0 to 3.0% (w / w) of distarch phosphate.

[0162] Another preferred skin care composition comprises

[0163] (a) freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0164] (b) 0.05 to 25.0% (w / w) of dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) of dicaprylyl carbonate; and

[0165] (c) 0.05 to 5.0% (w / w) of tapioca starch, preferably 2.0 to 3.0% (w / w) of tapioca starch.

[0166] Another preferred skin care composition comprises

[0167] (a)Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0168] (b)0.05 to 25.0% (w / w) ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) ethylhexyl cocoate; and

[0169] (c)0.05 to 5.0% (w / w) tapioca starch, preferably 2.0 to 3.0% (w / w) tapioca starch.

[0170] Another preferred skin care composition comprises

[0171] (a)Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0172] (b)0.05 to 25.0% (w / w) dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) dicaprylyl carbonate; and

[0173] (c)0.01 to 2.5% (w / w) PEG-40 hydrogenated castor oil, preferably 0.5 to 1.0% (w / w) PEG-40 hydrogenated castor oil.

[0174] Another preferred skin care composition comprises

[0175] (a)Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0176] (b)0.05 to 25.0% (w / w) ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) ethylhexyl cocoate; and

[0177] (c)0.01 to 2.5% (w / w) PEG-40 hydrogenated castor oil, preferably 0.5 to 1.0% (w / w) PEG-40 hydrogenated castor oil.

[0178] Another preferred skin care composition comprises

[0179] (a)Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0180] (b)0.05 to 25.0% (w / w) of dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) of dicaprylyl carbonate; and

[0181] (c)0.01 to 2.0% (w / w) of tocopherol, preferably 0.1 to 1.0% (w / w) of tocopherol.

[0182] Another preferred skin care composition comprises

[0183] (a)Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0184] (b)0.05 to 25.0% (w / w) of ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) of ethylhexyl cocoate; and

[0185] (c)0.01 to 2.0% (w / w) of tocopherol, preferably 0.1 to 1.0% (w / w) of tocopherol.

[0186] Another preferred skin care composition comprises

[0187] (a)Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0188] (b)0.05 to 25.0% (w / w) of dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) of dicaprylyl carbonate; and

[0189] (c)0.01 to 2.0% (w / w) of tocopheryl acetate, preferably 0.1 to 1.0% (w / w) of tocopheryl acetate.

[0190] Another preferred skin care composition comprises

[0191] (a)Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0192] (b)0.05 to 25.0% (w / w) ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) ethylhexyl cocoate; and

[0193] (c)0.01 to 2.0% (w / w) tocopheryl acetate, preferably 0.1 to 1.0% (w / w) tocopheryl acetate.

[0194] Another preferred skin care composition comprises

[0195] (a)Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0196] (b)0.05 to 25.0% (w / w) dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) dicaprylyl carbonate; and

[0197] (c)0.5 to 20.0% (w / w) ethanol, preferably 8.0 to 15.0% (w / w) ethanol.

[0198] Another preferred skin care composition comprises

[0199] (a)Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0200] (b)0.05 to 25.0% (w / w) ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) ethylhexyl cocoate; and

[0201] (c)0.5 to 20.0% (w / w) ethanol, preferably 8.0 to 15.0% (w / w) ethanol.

[0202] Another preferred skin care composition comprises

[0203] (a) Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0204] (b) 0.05 to 25.0% (w / w) of dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) of dicaprylyl carbonate; and

[0205] (c) 0.05 to 0.5% (w / w) of phenoxyethanol, preferably 0.1 to 0.2% (w / w) of phenoxyethanol.

[0206] Another preferred skin care composition comprises

[0207] (a) Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0208] (b) 0.05 to 25.0% (w / w) of ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) of ethylhexyl cocoate; and

[0209] (c) 0.05 to 0.5% (w / w) of phenoxyethanol, preferably 0.1 to 0.2% (w / w) of phenoxyethanol.

[0210] Another preferred skin care composition comprises

[0211] (a) Lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably lyophilized or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0212] (b) 0.05 to 25.0% (w / w) of dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) of dicaprylyl carbonate; and

[0213] (c) 0.05 to 0.5% (w / w) of octanediol, preferably 0.1 to 0.2% (w / w) of octanediol.

[0214] Another preferred skin care composition comprises

[0215] (a) Freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0216] (b) 0.05 to 25.0% (w / w) ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) ethylhexyl cocoate; and

[0217] (c) 0.05 to 0.5% (w / w) octanediol, preferably 0.1 to 0.2% (w / w) octanediol.

[0218] Another preferred skin care composition comprises

[0219] (a) Freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0220] (b) 0.05 to 25.0% (w / w) dicaprylyl carbonate, preferably 7.5 to 10.0% (w / w) dicaprylyl carbonate; and

[0221] (c) 0.05 to 5.0% (w / w) methylpropanediol, preferably 1.0 to 5.0% (w / w) methylpropanediol.

[0222] Another preferred skin care composition comprises

[0223] (a) Freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes strain, preferably freeze-dried or spray-dried live bacteria of at least one Propionibacterium acnes SLST C3 strain and at least one Propionibacterium acnes SLST K8 strain;

[0224] (b) 0.05 to 25.0% (w / w) ethylhexyl cocoate, preferably 7.5 to 10.0% (w / w) ethylhexyl cocoate; and

[0225] (c) 0.05 to 5.0% (w / w) methylpropanediol, preferably 1.0 to 5.0% (w / w) methylpropanediol.

[0226] Cosmetic and therapeutic uses

[0227] The skin care compositions described above can be used to modulate the skin microbiome, particularly for maintaining healthy skin, such as skin without acne. Compositions comprising at least one live bacterial strain of Propionibacterium acnes can help the skin reverse a microbiome disease state to a healthy microbiome state. In a preferred embodiment, the skin care composition is used for preventing acne formation or treating acne. In another preferred embodiment, the skin care composition is used for preventing acne recurrence in a subject who has received standard acne treatment. Particularly preferably, the subject is a human.

[0228] In one aspect, the present invention provides a skin care composition as described above for use in a method of improving the skin appearance of a subject and / or modulating (such as increasing or decreasing) sebum production of the subject's skin cells and / or maintaining the subject's healthy skin, such as skin without acne. In another aspect, the present invention relates to the use of a skin care composition as described above for improving the skin appearance of a subject and / or modulating (such as increasing or decreasing) sebum production of the subject's skin cells and / or maintaining the subject's healthy skin, such as skin without acne. In one aspect, the present invention provides a skin care composition as described above for use in a method of treating or preventing a disorder selected from acne, oily skin, progressive macular hypomelanosis, dandruff, atopic eczema, atopic dermatitis, and rosacea in a subject. Particularly preferably, the subject is a human.

[0229] Also provided are methods of treating a subject's skin by administering a skin care composition as described above. These methods can be cosmetic or therapeutic methods. In one aspect, a method of improving the skin appearance of a subject and / or modulating sebum production of the subject's skin cells and / or maintaining the subject's healthy skin is provided, the method comprising topically administering a skin care composition as described above. In another aspect, the present invention provides a method of treating or preventing a disorder selected from acne, oily skin, progressive macular hypomelanosis, dandruff, atopic eczema, atopic dermatitis, and rosacea in a subject, the method comprising topically administering a skin care composition as described above. Particularly preferably, the subject is a human.

[0230] When the composition is applied to the skin, the amount of the composition applied to the skin is preferably between 0.5 g and 2.0 g, more preferably between 0.5 g and 1.0 g. In other words, the amount of the composition can correspond to at least 1.0 x10 5 CFU, at least 1.0 x 10 6 CFU, at least 1.0 x 10 7 CFU, at least 1.5 x 10 7 CFU, at least 2.0 x 10 7 CFU or at least 2.5 x 107 CFU.

[0231] Ready-to-use composition and kit of medicaments

[0232] The skin care composition of the present invention can be provided as a ready-to-use composition suitable for direct topical administration to the skin. Such compositions can be provided in different forms, including but not limited to gels, creams, lotions, ointments, pastes, soft pastes, suspensions, solutions, salves, waxes, milks, emulsions, etc. In such compositions, the lyophilized or spray-dried live bacteria are present in a mixture with other cosmetic or pharmaceutical excipients described elsewhere herein, such as emollients, fillers, etc. After applying these compositions to the skin, the dried bacteria are reactivated on the skin of the subject to whom the product is applied. The growth of the reactivated bacteria from the skin care composition will positively affect the microbial flora on the skin of the subject.

[0233] These ready-to-use compositions are preferably stable at room temperature for at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks, at least 11 weeks, at least 12 weeks, at least 13 weeks, at least 14 weeks, at least 15 weeks, at least 16 weeks, at least 17 weeks, at least 18 weeks, at least 19 weeks, at least 20 weeks, at least 21 weeks, at least 22 weeks, at least 23 weeks, at least 24 weeks, at least 25 weeks, at least 26 weeks, at least 27 weeks, at least 28 weeks, at least 29 weeks, at least 30 weeks or more than 30 weeks. In other words, such compositions are preferably stable at room temperature for at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months or more than 6 months. As used herein, a composition is considered stable if the reduction in the number of colony forming units present in the composition after storage is less than 3 log reduction, preferably less than 2 log reduction, more preferably less than 1 log reduction. In other words, if the reduction in the number of colony forming units present in the composition after storage relative to the number of colony forming units in the composition before storage is less than 1000-fold, preferably less than 100-fold, more preferably less than 10-fold, the composition is considered stable.

[0234] The skin care composition of the present invention can also be provided as a kit of medicaments, wherein the lyophilized or spray-dried bacteria are spatially separated from other components such as cosmetic or therapeutic components. For example, the kit of medicaments can take the form of a package having two spatially separated compartments, wherein the first compartment contains the lyophilized or spray-dried bacteria and the second compartment contains a cosmetic preparation such as an aqueous cosmetic preparation. Before use, the contents of the two compartments are mixed with each other by, for example, a consumer or a patient to provide a homogeneous skin care composition, which is then applied to the skin. The kit of medicaments assembly has the advantage that the bacteria can be kept in the lyophilized or spray-dried form before use, which is accompanied by a particularly high storage stability of the composition. In the kit of medicaments assembly, advantageously, the weight ratio of the bacteria in the first compartment to the cosmetic preparation, particularly the aqueous cosmetic preparation, in the second compartment is from 1:10 to 1:100, for example 1:10, 1:20, 1:30, 1:40, 1:50, 1:60, 1:70, 1:80, 1:90 or 1:100. After mixing the contents of the two compartments, the skin care composition preferably contains 1-10% by weight of the lyophilized or spray-dried bacteria and 99-90% by weight of a cosmetic preparation such as an aqueous cosmetic preparation. According to the present invention, the kit of medicaments can be provided in a two-compartment system described, for example, in Lyo-Ject® two-compartment syringe, V-LK® two-compartment carpu and WO2018077598 A1. In another preferred embodiment, the lyophilized or spray-dried bacteria in the first compartment can be suspended in a lipid or an oil. This will greatly facilitate packaging and filling. In addition, the surrounding lipid or oil will protect the bacteria from premature rehydration. Preferably, the bacteria are suspended in ethylhexyl cocoate or dicaprylyl carbonate. The weight ratio of the bacteria to the oil or lipid is preferably between 1:1 and 1:2.

[0235] In one embodiment, the skin care composition of the present invention is an aqueous preparation such as a gel. The aqueous preparation as meant herein encompasses aqueous solutions as well as aqueous dispersions. In one embodiment, the skin care composition is an oil-in-water emulsion. If the skin care composition contains an oil phase, for example when using an oil-in-water emulsion, preferably the oil phase contains triglycerides and / or octyldodecanol. In addition, the oil phase may contain one or more oils selected from lecithin, olive oil, sunflower oil, jojoba oil, soybean oil, peanut oil, rapeseed oil, almond oil, palm oil, coconut oil, castor oil, wheat germ oil, grape seed oil, safflower oil, evening primrose oil, macadamia oil and the like.

[0236] When used in the context of a method or composition, the term "comprising" means that other method steps or composition components may be present in addition to the recited method steps or components. The use of the term "comprising" indicates inclusion rather than limitation. For example, a composition "comprising" components A + B may also contain C as an additional component. Similarly, a method "including" steps (a) and (b) may also include (c) as an additional method step. In contrast, when used in the context of a method or composition, the term "consisting of" means that the method or composition does not include any other method steps or composition components not recited in the description of the corresponding composition or method. For example, a composition consisting of components A + B is limited to these two components and does not contain any other components other than A and B. Similarly, a method consisting of steps (a) and (b) is a two-step method and does not contain any other method steps other than (a) and (b). However, it should be understood that any method or composition described herein as "including" certain method steps or components may preferably consist essentially of the recited method steps or components, or more preferably may consist of the recited method steps or components. Additionally, unless the context otherwise requires, singular terms shall include the plural, and plural terms shall include the singular.

[0237] Literature

[0238] Bek-Thomsen, M., Lomholt, H.B. and Kilian, M. (2008), Acne is not associated with yet-uncultured bacteria, J. Clin. Microbiol. 46, 3355—3360.

[0239] Belkaid, Y. and Segre, J.A. (2014), Dialogue between skin microbiota and immunity, Science 346, 954-959.

[0240] Downing, D.T., Stewart, M.E., Wertz, P.W. and Strauss, J.S. (1986), Essential fatty acids and acne, J. Am. Acad. Dermatol. 14, 221-225.

[0241] Fitz-Gibbon, S., Tomida, S., Chiu, B.-H., Nguyen, L., Du, C., Liu, M., Elashoff, D., Erfe, M.C., Loncaric, A., Kim, J., et al. (2013), Propionibacterium acnes strain populations in the human skin microbiome associated with acne, J. Invest. Dermatol. 133, 2152-2160.

[0242] Grice, E.A. and Segre, J.A. (2011), The skin microbiome, Nat. Rev. Microbiol. 9, 244-253.

[0243] Holmes, AD. (2013), Potential role of microorganisms in the pathogenesis of rosacea, J. Am. Acad. Dermatol. 69, 1025-1032.

[0244] Kong, H.H., Oh, J., Deming, C., Conlan, S., Grice, E.A., Beatson, M.A., Nomicos, E., Polley, E.C., Komarow, H.D., Murray, P.R., et al. (2012), Temporal shifts in the skin microbiome associated with disease flares and treatment in children with atopic dermatitis, Genome Res. 22, 850-859.

[0245] Letawe, C., Boone, M. and Piérard, G.E., (1998), Digital image analysis of the effect of topically applied linoleic acid on acne microcomedones, Clin. Exp. Dermatol. 23, 56-58.

[0246] Lomholt, H.B. and Kilian, M., (2010), Population Genetic Analysis of Propionibacterium acnes Identifies a Subpopulation and Epidemic Clones Associated with Acne, PLoS ONE 5.

[0247] McDowell, A., Barnard, E., Nagy, I., Gao, A., Tomida, S., Li, H., Eady, A., Cove, J., Nord, C.E. and Patrick, S., (2012), An Expanded Multilocus Sequence Typing Scheme for Propionibacterium acnes: Investigation of “Pathogenic”, “Commensal” and Antibiotic Resistant Strains, PLoS ONE 7, e41480.

[0248] Oh, J., Byrd, A.L., Deming, C., Conlan, S., NISC Comparative Sequencing Program, Kong, H.H. and Segre, J.A., (2014), Biogeography and individuality shape function in the human skin metagenome, Nature 514, 59-64.

[0249] Rosson, R.A., Grund, A.D., Deng, M.-D. and Sanchez-Riera, F., (2004), Linoleate isomerase.

[0250] Scholz, C.F.P., Jensen, A., Lomholt, H.B., Brüggemann, H. and Kilian, M., (2014), A Novel High-Resolution Single Locus Sequence Typing Scheme for Mixed Populations of Propionibacterium acnes In Vivo, PLoS ONE 9, e104199.

[0251] Examples

[0252] The present invention is further illustrated by the following examples, which should in no way be construed as further limiting. The entire contents of all references cited throughout this application (including literature references, issued patents, published patent applications, and co-pending patent applications) are hereby expressly incorporated by reference.

[0253] Example 1: Evaluation of the minimum inhibitory concentration (MIC)

[0254] In microbiology, the MIC is the lowest concentration of a chemical that prevents visible growth of bacteria. The resulting MIC value depends on both the microorganism and the test compound. The MIC is determined by preparing a series of concentrations of the test compound in vitro, incubating the solutions with separate batches of cultured bacteria, and measuring the results using agar dilution or broth microdilution. For experiments using Propionibacterium acnes strains C3 and K8, a medium containing yeast extract, soy peptone, and dextrose is used. The MIC is determined by turbidity.

[0255] A total of 125 cosmetic excipients were tested for compatibility with Propionibacterium acnes strains C3 and K8. Hydrophilic compounds were tested in the form of 96-well plate liquid broth, poorly soluble compounds were pre-dissolved in DMSO before dilution in liquid broth, and hydrophobic or solid compounds were tested in an agar well diffusion test assay. Different dilutions of the test excipients in the medium were prepared in Falcon tubes and then dispensed into deep well plates using a multi-channel pipettor. Each well received 1.2 ml and was subsequently inoculated with 120 μl of a Propionibacterium acnes bacterial suspension (strains C3 and K8). 200 μl was transferred from the deep well plate to each well of a transparent 96-well parallel sample plate. The remainder was discarded. The plates were then incubated anaerobically at 37 °C for 3 days. After incubation, the test plates were visually inspected and scored for growth inhibition or complete growth inhibition of the wells to determine the minimum inhibitory concentration.

[0256] Results:

[0257] It has been found that the compatibility of standard excipients with bacterial strains varies widely. Only a few excipients do not cause inhibition or cause significant inhibition. The following table lists these excipients and their compatibility values with Propionibacterium acnes strains C3 and K8. The compatibility values represent the MIC values of the listed ingredients when using the tested Propionibacterium acnes strains. At this concentration [% w / vol], the test compound begins to interfere with the growth of the test strain. The compound can be used in formulations at concentrations below this concentration.

[0258]

[0259] Example 2: Reactivation assay of freeze-dried bacteria in the prototype formulation

[0260] Based on the results from the MIC tests, a set of 15 prototype formulations was designed to test the efficient reactivation of freeze-dried Propionibacterium acnes bacteria. For this purpose, freeze-dried powders of Propionibacterium acnes strains K8 and C3 were produced and mixed with the other ingredients shown below. The ingredients are listed in % (w / w). The formulations were provided in two pre-mixed compositions A and B, which were ultimately mixed with each other. The following formulations were manufactured:

[0261]

[0262]

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270]

[0271]

[0272]

[0273]

[0274]

[0275]

[0276] The formulations are depicted in tabular form in Figure 1 For formulations 1 - 11, the CFU target value is 1x10 7 . For formulations 12 - 15, the CFU target value is 1x10 6 . The CFU count is performed directly after mixing the formulations to evaluate the effect of the prototype formulations on the resuscitation and subsequent stabilization rate of the bacteria. Briefly, formulation component A (lyophilisate / oil suspension) and formulation component B (hydrogel / aqueous solution) are mixed together and vortexed thoroughly. Immediately after mixing, aliquots are taken and transferred to 0.9% NaCl. Further dilutions are plated on COST agar plates and incubated anaerobically at 37°C for 5 days.

[0277] Results :

[0278] The results are depicted in Figure 2 and 3 It can be seen that all formulations 1 - 15 showed acceptable reactivation of the freeze - dried bacteria. The components had no negative impact on the bacterial reactivation.

[0279] Example 3: Reactivation assay of freeze-dried bacteria in the prototype formulation

[0280] Similar to the formulations tested in Example 2, another set of 11 prototype formulations with a higher concentration of the excipients of the present invention were prepared. The effect of the formulations on the reactivation of freeze-dried Propionibacterium acnes bacteria was tested as described in Example 2. As a control, a formulation containing the lyophilizate and a small amount of excipient was prepared. The following formulations were manufactured:

[0281]

[0282]

[0283]

[0284]

[0285]

[0286]

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293] The formulations are depicted in tabular form in Figure 4 . For all formulations, the CFU target value was 1x10 9 . CFU counts were performed immediately after mixing the formulations and 1 hour later as described in Example 2.

[0294] Results :

[0295] The results are depicted in Figure 5 . It can be seen that all formulations 1 - 11 showed acceptable reactivation of the freeze-dried bacteria. Even at the high concentrations used in this experiment, the components had no negative effect on the reactivation of the bacteria.

[0296] Example 4: Comparative reactivation assay

[0297] To compare the excipients identified herein as being particularly useful for compositions containing freeze-dried or spray-dried live bacteria with other excipients having the same or similar functions, comparative formulations V-1 to V-10 were prepared and their effect on bacterial reactivation was tested as described in Example 2. As a positive control, a formulation containing a freeze-dried product that had been found to have no negative impact on bacterial reactivation was used. The following formulations were manufactured for comparison:

[0298]

[0299]

[0300]

[0301]

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309] The formulations are depicted in tabular form in Figure 7 . For all formulations, the CFU target value was 1x10 9 . The CFU count was performed immediately after mixing the formulations as described in Example 2 above, and also after 1 hour or 6 hours respectively.

[0310] Results :

[0311] The results are depicted in Figure 6 . It can be seen that formulations V-1 to V-10 have a significant negative impact on the reactivation of bacteria. This confirms that a large number of excipients commonly used in cosmetic formulations negatively interfere with the viability or reactivation of the freeze-dried bacteria used in this study.

Claims

1. A skin care composition for topical administration to the skin, comprising: (a) Freeze-dried or spray-dried live bacteria of at least one Cutibacterium acnes SLST C3 strain and / or at least one Cutibacterium acnes SLST K8 strain, wherein each Cutibacterium acnes strain in the composition is present in an amount of 10 4 -10 11 CFU / ml; and (b) an emollient selected from dicaprylyl carbonate, ethylhexyl cocoate, and mixtures thereof, wherein the dicaprylyl carbonate and / or the ethylhexyl cocoate is present in the skin care composition in an amount of 7.5 to 20.0% (w / w).

2. The skin care composition according to claim 1, wherein the emollient is dicaprylyl carbonate.

3. The skin care composition according to claim 1, wherein the emollient is ethylhexyl cocoate.

4. The skin care composition according to claim 1, wherein the emollient is a mixture of dicaprylyl carbonate and ethylhexyl cocoate.

5. The skin care composition according to claim 4, wherein the dicaprylyl carbonate and the ethylhexyl cocoate are present in equal amounts.

6. The skin care composition according to claim 4, wherein the total amount of the emollient in the composition does not exceed 20.0% (w / w).

7. The skin care composition according to any one of claims 1-6, wherein the composition does not contain any other emollient other than dicaprylyl carbonate and / or ethylhexyl cocoate.

8. The skin care composition according to any one of claims 1-6, wherein the composition further comprises a thickening agent selected from Chondrus crispus extract, hydroxypropyl starch phosphate, and mixtures thereof.

9. The skin care composition according to any one of claims 1-6, wherein the composition further comprises a filler selected from distarch phosphate, tapioca starch, and mixtures thereof.

10. The skin care composition according to any one of claims 1-6, wherein the composition further comprises an antioxidant selected from tocopherol, tocopheryl acetate, and mixtures thereof.

11. The skin care composition according to any one of claims 1-6, wherein the composition further comprises a preservative selected from ethanol, phenoxyethanol, octanediol, methylpropanediol, and mixtures thereof.

12. The skin care composition according to any one of claims 1-6, wherein the composition further comprises PEG-40 hydrogenated castor oil as a solubilizer.

13. The skin care composition according to any one of claims 1-6, wherein the composition further comprises a citric acid / citrate buffer as a pH regulator.

14. The skin care composition according to any one of claims 1-6, wherein the composition further comprises at least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes strain SLST A5.

15. The skin care composition according to any one of claims 1-6, wherein the composition further comprises at least one freeze-dried or spray-dried live bacterium of Propionibacterium acnes strain SLST F4.

16. The skin care composition according to any one of claims 1-6, wherein the concentration of each Propionibacterium acnes strain is at least 0.5% (w / v) of the skin care composition.

17. The skin care composition according to claim 1, wherein in the composition, the at least one Propionibacterium acnes SLST C3 type strain and the at least one Propionibacterium acnes SLST K8 type strain have equal concentrations.

18. The skin care composition according to claim 1, wherein in the composition, the at least one Propionibacterium acnes SLST C3 type strain is present at a higher concentration than the at least one Propionibacterium acnes SLST K8 type strain, or wherein in the composition, the at least one Propionibacterium acnes SLST K8 type strain is present at a higher concentration than the at least one Propionibacterium acnes SLST C3 type strain.

19. The skin care composition according to any one of claims 1-6, wherein each Propionibacterium acnes strain in the composition is present in an amount of 10 7 -10 10 CFU / ml.

20. The skin care composition according to any one of claims 1-6, wherein the total amount of bacteria in the composition is 10 4 -10 11 CFU / ml.

21. The skin care composition according to any one of claims 1-6, wherein the total amount of bacteria in the composition is 10 7 -10 10 CFU / ml.

22. The skin care composition according to any one of claims 1 - 6, wherein the composition is in the form of a gel, cream, ointment or lotion.

23. The skin care composition according to any one of claims 1 - 6, wherein the composition is used in a method for improving the skin appearance of a subject and / or regulating sebum production of the subject's skin cells and / or maintaining the healthy skin of the subject.

24. The skin care composition according to any one of claims 1 - 6, wherein the composition is used in a method for treating or preventing a disorder selected from acne, oily skin, progressive macular hypomelanosis, dandruff, atopic eczema, atopic dermatitis and rosacea in a subject.

25. Use of the skin care composition according to any one of claims 1 - 22 in the preparation of a medicament for improving the skin appearance of a subject and / or regulating sebum production of the subject's skin cells and / or maintaining the healthy skin of the subject.

26. Use of the skin care composition according to any one of claims 1 - 22 in the preparation of a medicament for treating or preventing a disorder selected from acne, oily skin, progressive macular hypomelanosis, dandruff, atopic eczema, atopic dermatitis and rosacea in a subject.

27. The skin care composition according to claim 23, wherein the subject is human.

28. The skin care composition according to claim 24, wherein the subject is human.

29. The use according to claim 25 or 26, wherein the subject is human.

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