Novel compositions for anti-aging treatment of the skin

Abstract

This invention relates to the treatment and / or prevention of skin aging. More specifically, this invention relates to the administration of live microorganisms to improve the appearance of the skin and reduce the signs of skin aging.

Description

[Technical Field]

[0001] The present invention relates to compositions and methods for treating skin aging, particularly topical compositions and methods of use. The present invention relates to topical compositions comprising viable microorganisms. In particular, the present invention relates to compositions comprising live probiotic microorganisms for improving the appearance of the skin and reducing signs of skin aging, and to methods of topical use of such compositions.

[0002] In particular, the present invention relates to a novel composition comprising viable microorganisms that exhibit active metabolism on the skin. [Background technology]

[0003] This invention relates to compositions for preventing or treating skin aging or photodamage, the use of said compositions, and methods of use of said compositions. The compositions contain live probiotic microorganisms that can reverse the signs of aging skin.

[0004] During aging, the skin undergoes significant clinical and morphological changes. The skin ages physiologically due to both intrinsic and extrinsic factors, known as chronological aging and photoaging, respectively. Intrinsic factors are driven by telomere shortening and cellular senescence, while extrinsic factors such as ultraviolet radiation and pollution contribute to the production of reactive oxygen species (ROS) and free radicals.

[0005] Aging is a gradual deterioration that occurs as we age and affects everyone. One of the most visually noticeable consequences of aging can be seen in the skin. The skin loses its firmness and elasticity, resulting in sagging, enlarged pores, and wrinkles. In addition, the rate of skin cell turnover decreases, the skin loses its luster and appears rough, the skin becomes dry as a result of decreased sebaceous gland activity, and the number of pigmented spots increases.

[0006] Skin aging can be considered at several levels, including phenotypic (as described above), histologically, and / or molecularly. Histologically, the junction between the dermis and epidermis flattens, and melanocyte density decreases. Dermal thickness decreases, and simultaneously, angiogenesis and the number of both mast cells and fibroblasts decrease. The decrease in skin elasticity and firmness is a result of changes in the extracellular matrix (ECM) of the dermis, such as changes in the production and location of collagen and elastin fibers. Glandular activity also decreases, resulting in less sebum, and subsequently, the skin becomes even drier. Cellularly, aging occurs due to the accumulation of senescent cells, changes in the cell cycle, and accumulation of free radicals / oxidative stress, as well as changes in glycation patterns, particularly the glycation pattern of collagen. Signs of aging such as dryness, rough texture, pigmentation, and wrinkles, as well as a decrease in the skin barrier and changes in biomechanical function, are the main histological changes.

[0007] As the Westernized population ages, treatments that can prevent and reverse the signs of skin aging are attracting increasing attention due to their cosmetic and economic potential.

[0008] The age-related skin changes described above are caused by intrinsic and extrinsic factors. Intrinsic factors include time-dependent and genetic factors, while extrinsic factors are caused by environmental influences, such as sunlight (known as photoaging), pollution exposure, and smoking. In particular, sunlight exposure can lead to the generation of reactive oxygen species (ROS) and free radicals, which ultimately result in oxidative stress and subsequent cell damage. Furthermore, a recent study by Howard et al. (2022) found that while the diversity of the skin microbiome increased with age in all sampled areas, the abundance of Lactobacillus decreased, suggesting that the skin microbiome may also play a role in skin aging.

[0009] Aging is caused by a wide variety of factors that interact and influence each other in complex ways. These changes can not only be aesthetically unpleasant for some people, but can also lead to problems with protective barriers and subsequent medical issues. Especially in Westernized societies, as the population ages, there is a great deal of cosmetic and economic interest in reversing / preventing the appearance of signs of aging.

[0010] With the aging of the Westernized population, treatments that can prevent and reverse the signs of skin aging are attracting increasing attention due to their cosmetic and economic potential. The anti-aging effect on facial skin can be assessed by evaluating the thickness of the subepidermal hypoechoic band (SLEB), dermal density, skin firmness and elasticity, skin moisture content, TEWL, crow's feet wrinkles, age spots, smoothness, and skin radiance, and comparing these to baseline measurements. SLEB has previously been shown to quantify skin aging by its thickness (Gniadecka et al., 1994). Therefore, SLEB has been shown to function well in quantifying aging, particularly in sun-exposed areas such as the cheek region.

[0011] The correlation between skin lipids and bacterial genus may be important for age-dependent changes in the skin microbiome. In this invention, viable lactic acid bacteria producing antimicrobial metabolites and organic acids are topically applied in lipid formulations to modulate the skin microbiome and reverse aging. When viable bacteriocin-producing microorganisms are applied, it has been shown that they can surprisingly improve the appearance of the skin and reduce signs of aging. Therefore, it is expected that the age-related decrease in Lactobacillus also leads to a decrease in bacteriocin-producing Lactobacillus, thereby enabling an increase in the diversity of the skin microbiome. Thus, applying viable Lactobacillus to aging skin increases the abundance of Lactobacillus in the microbiome, and through metabolism on the skin, the metabolites produced by these Lactobacillus reduce the diversity of the microbiome to reflect a younger skin microbiome. [Overview of the project]

[0012] The present invention relates to a composition for topical use containing viable isolated microorganisms, or a method of using viable isolated microorganisms for treating or preventing skin aging.

[0013] In one aspect of the present invention, the composition is for treating or preventing skin aging measurable as a decrease in at least one of the following parameters: wrinkles, pigmentation, thickness of the subcutaneous hypoechoic band (SLEB), or transepidermal water loss (TEWL).

[0014] Or, as an increase in at least one of the following parameters: dermal density, skin moisture content, skin elasticity, skin gloss or smoothness.

[0015] Another aspect of the present invention is a method of skin treatment for treating, alleviating, suppressing, preventing and / or precluding skin aging.

[0016] The present invention is based on multiple mechanisms of action caused by viable species of lactic acid bacteria that can reduce signs of aging.

[0017] One aspect of the present invention relates to a composition for use in treating, alleviating, suppressing, preventing and / or precluding skin aging, comprising at least one viable isolated microorganism.

[0018] Another aspect of the present invention is that the composition comprises at least one viable isolated microorganism, (a) a reduction of at least 5% in wrinkles, (b) a reduction of at least 5% in the density of pigmented spots, and / or (c) an increase in firmness measured as a reduction in the suction distance of more than 5%, and relates to a composition for use in treating, alleviating, suppressing, preventing and / or precluding skin aging.

[0019] A further aspect of the present invention relates to a composition for use in the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, comprising at least two live lactic acid bacteria for use in topical treatment of the skin, which provides an increase of at least 5% in dermal density after 28 days of daily topical treatment.

[0020] Further aspects of the present invention include a composition comprising at least one living isolated microorganism, (a) a reduction of at least 2% in the thickness of the subepidermal hypoechoic band (SLEB), and / or (b) relating to a composition for use in the treatment, mitigation, inhibition, prevention and / or prevention of skin aging by providing a reduction of at least 2% in TEWL.

[0021] Furthermore, in aspects of the present invention, the composition comprises at least one living isolated microorganism. (a) an increase in skin tone of more than 10%, and / or (b) relating to compositions for use in treating, mitigating, inhibiting, preventing and / or preventing skin aging by providing an increase in smoothness of more than 10%.

[0022] A further aspect of the present invention relates to a method of skin treatment comprising administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days for the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, the method being (a) A reduction of at least 5% in wrinkles, (b) A reduction of at least 5% in pigmented spot density, and / or (c) Provides increased tension, which is measured as a reduction in suction distance of more than 5%.

[0023] A further aspect of the present invention relates to a method of skin treatment comprising administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days for the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, the method being (a) an increase of at least 5% in dermal density, (b) an increase in skin moisture content of more than 10%, and / or (c) Provides an increase in skin elasticity of more than 2%.

[0024] A further aspect of the present invention relates to a method of skin treatment comprising administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days for the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, the method being (a) a reduction of at least 2% in the thickness of the subepidermal hypoechoic band (SLEB), and / or (b) Provides at least a 2% reduction in TEWL.

[0025] A further aspect of the present invention relates to a method of skin treatment comprising administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days for the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, the method being (a) an increase in skin tone of more than 10%, and / or (b) Provides a smoothness increase of more than 10%.

[0026] A further aspect of the present invention relates to live lactic acid bacteria for use in topical skin treatment, which yield at least a 2% reduction in the thickness of subepidermal hypoechoic bands (SLEBs) after 28 days of daily topical treatment.

[0027] Furthermore, an aspect of the present invention is a live lactic acid bacterium for use in topical treatment of the skin, wherein the lactic acid bacterium is present after daily topical treatment for 28 days. (a) an increase of at least 5% in dermal density, (b) an increase in skin moisture content of more than 10%, and / or (c) Regarding live lactic acid bacteria that provide an increase in skin elasticity of more than 2%.

[0028] The present invention will be described in more detail below. [Modes for carrying out the invention]

[0029] definition Before discussing the present invention in further detail, the following terms and conventions are first defined. The term "skin aging" refers to measurable signs of skin aging that may be visible and / or measurable through clinical evaluation. Typical signs of skin aging include wrinkles, sagging, hyperpigmentation, or enlarged pores, and the skin may appear dull, rough, and dry.

[0030] Efficacy is measured as an improvement in at least one of the following parameters: SLEB thickness (μm), dermal density (pixels / mm²). 2 ), skin firmness (R0) (μm), skin elasticity (R2) (%), skin moisture content (AU), TEWL (g / h / m²) 2 ), crow's feet score, blemish score, skin smoothness score, or skin radiance score.

[0031] In a preferred embodiment of the present invention, anti-aging improvement is measured as a reduction in SLEB, where the thickness of the SLEB decreases by at least 2%, at least 3%, at least 4%, at least 5%, or at least 6% after 56 days.

[0032] In a preferred embodiment of the present invention, anti-aging improvement is achieved by reducing dermal density (pixels / mm²). 2 Measured as an increase in ), dermal density increases by at least 5%, at least 10%, or at least 15% after 56 days.

[0033] In a preferred embodiment of the present invention, anti-aging improvement is measured as an increase in skin firmness, which is determined as a reduction of at least 5%, at least 8%, at least 10%, or at least 12% in the suction distance (R0) (μm) after 56 days.

[0034] In preferred embodiments of the present invention, anti-aging improvement is measured as an increase in skin elasticity (R2) (%), where skin elasticity increases by at least 2%, at least 4%, or at least 6% after 56 days.

[0035] In a preferred embodiment of the present invention, anti-aging improvement is measured as an increase in skin moisture content (AU), where skin moisture content increases by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% after 56 days.

[0036] In a preferred embodiment of the present invention, anti-aging improvement is achieved by TEWL (g / h / m²). 2 Measured as a decrease in ), TEWL decreases by at least 2%, at least 3%, or at least 4% after 56 days.

[0037] In a preferred embodiment of the present invention, anti-aging improvement is measured as a percentage reduction in the crow's feet wrinkle score, where wrinkles are reduced by at least 10%, at least 15%, or at least 20% after 56 days.

[0038] In a preferred embodiment of the present invention, anti-aging improvement is measured as a decrease in the pigment spot score (%), and the pigment spot density decreases by at least 5%, at least 10%, at least 15%, at least 20%, or at least 25% after 56 days.

[0039] In a preferred embodiment of the present invention, anti-aging improvement is measured as an increase in the skin smoothness score, where the increase in the skin smoothness score is at least 10%, at least 15%, at least 20%, at least 25%, or at least 30% after 56 days.

[0040] In a preferred embodiment of the present invention, anti-aging improvement is measured as an increase in skin radiance score, where the increase in skin radiance score is at least 10%, at least 20%, at least 25%, at least 30%, or at least 40% after 56 days.

[0041] In a preferred embodiment of the present invention, the anti-aging improvement of the skin is measured with respect to at least two parameters selected from the following: SLEB thickness (μm), dermal density (pixels / mm²). 2) Skin firmness (R0) (μm), skin elasticity (R2) (%), skin moisture content (AU), TEWL (g / h / m 2 ) Wrinkling score at the outer corner of the eye, pigmentation score, skin smoothness score, or skin gloss score.

[0042] In a preferred embodiment of the present invention, the anti-aging improvement of the skin is measured for at least three parameters selected from the following: thickness of SLEB (μm), dermal density (pixels / mm 2 ) Skin firmness (R0) (μm), skin elasticity (R2) (%), skin moisture content (AU), TEWL (g / h / m 2 ) Wrinkling score at the outer corner of the eye, pigmentation score, skin smoothness score, or skin gloss score.

[0043] In a preferred embodiment of the present invention, the anti-aging improvement of the skin is measured for at least four parameters selected from the following: thickness of SLEB (μm), dermal density (pixels / mm 2 ) Skin firmness (R0) (μm), skin elasticity (R2) (%), skin moisture content (AU), TEWL (g / h / m 2 ) Wrinkling score at the outer corner of the eye, pigmentation score, skin smoothness score, or skin gloss score.

[0044] In a preferred embodiment of the present invention, the anti-aging improvement of the skin is measured for at least five parameters selected from the following: thickness of SLEB (μm), dermal density (pixels / mm 2 ) Skin firmness (R0) (μm), skin elasticity (R2) (%), skin moisture content (AU), TEWL (g / h / m 2 ) Wrinkling score at the outer corner of the eye, pigmentation score, skin smoothness score, or skin gloss score.

[0045] In a preferred embodiment of the present invention, the anti-aging improvement of the skin is measured for at least six parameters selected from the following: thickness of SLEB (μm), dermal density (pixels / mm 2 ) Skin firmness (R0) (μm), skin elasticity (R2) (%), skin moisture content (AU), TEWL (g / h / m2 ), crow's feet score, blemish score, skin smoothness score, or skin radiance score.

[0046] In a preferred embodiment of the present invention, the anti-aging improvement of the skin is measured with respect to at least seven parameters selected from the following: SLEB thickness (μm), dermal density (pixels / mm²). 2 ), skin firmness (R0) (μm), skin elasticity (R2) (%), skin moisture content (AU), TEWL (g / h / m²) 2 ), crow's feet score, blemish score, skin smoothness score, or skin radiance score.

[0047] In a preferred embodiment of the present invention, the anti-aging improvement of the skin is measured with respect to at least eight parameters selected from the following: SLEB thickness (μm), dermal density (pixels / mm²). 2 ), skin firmness (R0) (μm), skin elasticity (R2) (%), skin moisture content (AU), TEWL (g / h / m²) 2 ), crow's feet score, blemish score, skin smoothness score, or skin radiance score.

[0048] In a preferred embodiment of the present invention, the anti-aging improvement of the skin is measured with respect to at least nine parameters selected from the following: SLEB thickness (μm), dermal density (pixels / mm²). 2 ), skin firmness (R0) (μm), skin elasticity (R2) (%), skin moisture content (AU), TEWL (g / h / m²) 2 ), crow's feet score, blemish score, skin smoothness score, or skin radiance score.

[0049] In a preferred embodiment of the present invention, the anti-aging improvement of the skin is measured with respect to all of the following parameters: SLEB thickness (μm), dermal density (pixels / mm²) 2 ), skin firmness (R0) (μm), skin elasticity (R2) (%), skin moisture content (AU), TEWL (g / h / m²) 2 ), crow's feet score, blemish score, skin smoothness score, and skin radiance score.

[0050] The mechanism of action (MoA) of anti-aging in locally applied microorganisms is not yet fully understood, but several hypotheses are possible. These include, but are not limited to, local immunomodulation, ECM modulation, and direct cellular (keratinocyte) stimulation that leads to a reduction in the amount and systemic effects of aging, as well as increased collagen and elastin, ultimately resulting in proper ECM construction and structure. This may be mediated via mammalian mTOR signaling and antioxidant effects that alleviate oxidative stress that accumulates with age, but other modes of action are entirely possible. Moisturizing effects reduce cellular stress and improve barrier function. mTOR signaling is targeted by the bacterial metabolite rapamycin, which is known to have anti-aging effects. This signaling pathway is also intricately interconnected with immune signaling, which means that in the case of MoA, all of the above effects may be possible simultaneously, indicating multiple modes of action.

[0051] Furthermore, the regulation of the microbiome may influence aging parameters. The amount of Lactobacillus species decreases with age. This may be an age-related change in physiological function that later leads to changes in the microbiome, but perhaps the microbiome can regulate the host's physiological functions, thereby providing a more comfortable environment for them and enabling colonization. Therefore, topical application of live microorganisms to the skin may play a regulatory role in aging, further confirming that the regulation of the microbiome can influence aging.

[0052] Increased inflammation is associated with aging, a phenomenon known as "inflammaging." The applied microorganisms can alleviate this age-related inflammation by acting as immunostimulants, resulting in a skin phenotype that more closely resembles that of younger skin.

[0053] Another possible effect of MoA is that it restores the skin's acidic pH, alleviates oxidative stress, reduces and even reverses photoaged skin, and improves the skin's barrier function. By combining these effects with its effects on skin cells, it can act as a regulator of the skin microbiome, thereby indirectly improving the appearance of the skin.

[0054] This invention relates to the local application of live microorganisms.

[0055] Preferred microorganisms are, in particular, bacteria. Probiotic bacteria are preferably Lactiplantibacillus plantarum, Lactococcus lactis, Lacticaseibacillus rhamnosus, Lactobacillus helveticus, Lactobacillus jensenii, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus amylovorus, Lactobacillus amylolyticus, and Lactobacillus alimentarius. Lactobacillus alimentarius, Lactobacillus aviaries, Lactobacillus delbrueckii, Lactobacillus diolivorans, Lactobacillus farciminis, Lactobacillus gallinarum, Lacticaseibacillus casei, Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus hilgardii, Lactobacillus kephyranofachens Lactobacillus kefiri, Lactobacillus mucosaeLactobacillus mucosae), Lactobacillus panis, Lactiplantibacillus paraplantarum, Lactobacillus pontis, Latilactobacillus sakei, Lactobacillus saliverius, Lactobacillus sanfraciscensis, Lacticaseibacillus paracasei, Lactobacillus pentosus, Lactobacillus cellobiosus, Lactobacillus collinoides, Lactobacillus coliniformis (Lactobacillus Lactobacillus coryniformis, Lactobacillus curvatus, Levilactobacillus brevis, Lactobacillus buchneri, Lactobacillus fructivorans, Lactobacillus hilgardii, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus ingluviei, Weissella viridescens, Bifidobacterium bifidum Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifidobacterium breve, Bifidobacterium longumlongum), Bifidobacterium animalis, Carnobacterium divergens, Corynebacterium glutamicum, Leuconostoc citreum, Leuconostoc lactis, Leuconostoc mesenteroides, Leuconostoc pseudomesenteroides, Oenococcus oeni, Pasteuria nishizawae, Pediococcus acidilactici acidilactici), Pediococcus dextrinicus, Pediococcus parvulus, Pediococcus pentosaceus, Probionibacterium freudenreichii, Probionibacterium acidipropoinici, Enterococcus faecium, Enterococcus faecalis, Streptococcus thermophilus, Bacillus amyloricephaciens amyloliquefaciens), Bacillus atrophaeus, Bacillus clausii, Bacillus coagulans, Bacillus flexus, Bacillus fusiformis, Bacillus lentus (BacillusThe group is selected from Bacillus lentus, Bacillus licheniformis, Bacillus mega-terium, Bacillus mojavensis, Bacillus pumilus, Bacillus smithii, Bacillus subtilis, Bacillus vallismortis, Geobacillus stearothermophilus, or their variants.

[0056] In another aspect of the present invention, the probiotic microorganisms include Lactobacillus, Lactiplantobacillus, Horzaferia, Amyloractobacillus, Bombilactobacillus, Companilactobacillus, Rapidilactobacillus, Agrilactobacillus, Schleyferylactobacillus, Lougolactobacillus, Lactcaseibacillus, Lactilactobacillus, Delagrioa, Lycorolactobacillus, Lydilactobacillus, Furfurylactobacillus, Pausylactobacillus The selection is made from genera associated with the natural healthy skin microbiome, including the genera Chillus, Limosilactobacillus, Fructilactobacillus, Acetilactobacillus, Apiractobacillus, Reviractobacillus, Secandilactobacillus, Lentilactobacillus, Leuconostocpropionibacterium, Cucibacterium, Staphylococcus, Corynebacterium, Malassezia, Aspergillus, Cryptococcus, Rhodotorula, and / or Epicomum. In another preferred embodiment of the present invention, the probiotic strain is Staphylococcus epidermidis, Staphylococcus hominis, Cutibacterium acnes (Probionibacterium acnes), or any combination thereof.

[0057] In a preferred embodiment of the present invention, the probiotic strain is a Gram-positive bacterium.

[0058] In one preferred embodiment of the present invention, the composition is: Lactiplantibacillus plantarum LB356R (DSM 33094), Lactiplantibacillus plantarum LB244R (DSM 32996), Weissera viridesens LB10G (DSM 32906), Lacticaseibacillus paracasei LB113R (DSM 32907), Lacticaseibacillus paracasei LB116R (DSM 32908), Reviractobacillus brevis LB152G (DSM 32995), Lacticaseibacillus paracasei LB28R (DSM 32994), Enterococcus faecium LB276R (DSM 32997), Leuconostoc mesenteroides LB349R (DSM Lactobacillus plantarum LB316R (DSM 33091), Lactobacillus plantarum LB312R (DSM 33098), Pediococcus pentosaceus LB606R (DSM 33730), Lactobacillus plantarum LB679R (DSM 33731), Lactobacillus crispatus LB714R (DSM 33732), Lactobacillus gasseri LB905R (DSM 34094), Lactobacillus crispatus LB912R (DSM 34095), Lactobacillus crispatus LB919R (DSM 34097), Lactobacillus gensenii LB918R (DSM Includes at least one strain selected from the group consisting of 34096) and / or any mutant strains and / or any combination thereof.

[0059] "Live" microorganisms refer to "viable" organisms that can grow or metabolize. The "viability" of microorganisms is measured as colony-forming units (CFU). A "decrease" in microbial viability can be determined as the difference in CFU / g compared to the CFU / g when the composition was formulated.

[0060] Preferably, live microorganisms may be able to survive on the skin for at least 4 hours, for example, at least 6 hours, for example, at least 8 hours, for example, at least 10 hours, for example, at least 12 hours after topical application.

[0061] The microorganisms according to the present invention are preferably in isolated or purified form, where the term “isolated” means in particular that the microorganisms are cultured as a single culture and derived from a culture medium containing, for example, their natural culture medium. The term “purified” is not limited to absolute purity.

[0062] The microorganisms may, advantageously, exist in viable dried and / or spray-dried and / or freeze-dried and / or vacuum-dried crystalline forms.

[0063] In a preferred embodiment of the present invention, the probiotic strain is used as a live, isolated microorganism in a crystallized, dried form.

[0064] In a preferred embodiment of the present invention, the strain is used as a viable isolated strain dried into crystals of the cryoprotectant. The preferred form of crystals contains at least 10% cryoprotectant.

[0065] In one aspect of the present invention, the cryoprotectant is selected from salts, proteins, or polysaccharides.

[0066] In one aspect of the present invention, the cryoprotectant is selected from maltodextrin, trehalose, saccharose, lactose, mannitol, sucrose, glycerol, sorbitol, dextran, and inulin.

[0067] In one aspect of the present invention, the cryoprotectant composition comprises at least one of the following: maltodextrin, trehalose, saccharose, lactose, mannitol, sucrose, glycerol, sorbitol, dextran, and / or inulin.

[0068] Furthermore, it is preferable that microorganisms be present in the composition in an amount of 0.001% to 20% by weight, preferably 0.005% to 10% by weight, and particularly preferably 0.01% to 5% by weight.

[0069] A preferred embodiment of the present invention is approximately 1 × 10 per gram of composition. 3 ~1 × 10 14 CFU viable bacteria, more preferably about 1 × 10⁶ 4 ~1 × 10 10 CFU, most preferably about 1 × 10 5 ~1 × 10 9 This includes administration of viable bacteria to CFU.

[0070] In one preferred embodiment of the present invention, the dosage of live probiotic microorganisms in the composition is approximately 1 × 10⁶ per gram of composition. 4 CFU (Cell Fuel Units) or more, preferably about 1 × 10⁻⁶ 5 CFU / g or more, preferably about 1 × 10 5 These are viable bacteria with a CFU / g exceeding 100%.

[0071] "Lactic acid bacteria" include species from the Lactobacillus, Aerococcus, Bifidobacterium, Carnobacterium, Enterococcus, Leuconostoc, and Streptococci families. These are considered non-pathogenic and are generally used as probiotic bacteria to improve the gastrointestinal flora and in the treatment of gastrointestinal symptoms.

[0072] In one embodiment of the present invention, the preferred microorganism is an isolated wild-type lactic acid bacterium.

[0073] The term "bacteriocin" refers to antimicrobial peptides or proteins produced by bacteria that are active against microorganisms but do not harm the producing bacteria. For the purposes of the present invention, bacteriocins or bacteriocin sources generally include antimicrobial agents suitable for use in formulations as cosmetics or pharmaceuticals. Particularly preferred antimicrobial agents include "lanchbiotics" (i.e., polypeptides containing lanthionine and beta-methyllanthionine). Non-limiting examples of such lanchbiotics include nisin, e.g., nisin A or nisin Z, or nisin analogs or related lanthionine-containing peptides, e.g., pediosin, lactosin, lactasin (e.g., lactisin A, lactisin B, lactasin F), camosin, enterosin, plantarisin, subtilin, epidermine, cinnamycin, duramycin, ancovenin, Pep 5, etc. Other bacteriocins useful in the present invention include, for example, lanthobiotic and / or non-lanthobiotic bacteriocins, such as lactocossine (e.g., lactocossine A, lactocossine B, lactocossine M), leucocoin, helvetican, acidophilin, and caseisin, individually or in any combination.According to another embodiment of the teachings of the present invention, bacteriocins include niacin A, niacin Z, niacin Q, niacin F, niacin U, niacin U2, salivarcine X, lactisin J46, lactisin 481, lactisin 3147, salivarcine A, salivarcine A2, salivarcine A3, salivarcine A4, BHT-Aa, BHT Ab, Salivarsin A5, Salivarsin B, Streptin, Salivarsin Al, Streptin, Streptocosin A-FF22, Mutacin BNY266, Mutacin 1140, Mutacin K8, Mutacin II, SmbAB, Bovicin HJ50, Bovicin HC5, Macedosin, Leukosin C, Sakacin 5X, Enterosin CRL35 / Munditisin, Avidin A, Munditisin I, Enterosin HF, Babaricin A, Ubericin A, Leukosin A, Mesentericin Y105, Sakacin G, Curvasin A / Saka The group is selected from those containing Syn A, Lactosin 5, Cyclosin, Enterosin A, Giversin V41, Giversin M35, Babaricin, Coagulin, Pediosin PA-1, Munditisin, Pishikosin CS526, Pishikosin 126 / Vla, Sakasin, P-Carnobacteriocin BM1, Enterosin P, Pishikosin Vlb, Penosin A, Bacteriocin 31, Bacteriocin RC714, Hirasin JM79, Bacteriocin T8, Enterosin SE-K4, Carnobacteriocin B2, and Plantaricin.

[0074] The term "plantaricin" refers to bacteriocins derived from Lactiplantybacillus plantarum, and the main types of plantaricin include plantaricin A, plantaricin E, plantaricin F, plantaricin J, plantaricin K, plantaricin C, plantaricin D, plantaricin W, plantaricin T, and plantaricin S. Other plantaricins, such as plantaricin 35d, plantaricin MG, plantaricin 423, plantaricin 154, plantaricin 149, plantaricin 163, plantaricin LC74, plantaricin K25, plantaricin ST31, and plantaricin SA6, are also included. In particular, broad-spectrum plantaricins, such as plantaricin F, plantaricin DL3, plantaricin ZJ008, plantaricin MG, plantaricin Q7, plantaricin KL-1Y, plantaricin 163, and plantaricin 154.

[0075] In a preferred embodiment of the present invention, the bacteriocin is plantaricin. Plantaricin is produced by Lactiplantybacillus plantarum LB244R (DSM32996), Lactiplantybacillus plantarum LB356R (DSM33094), Lactiplantybacillus plantarum LB312R (DSM33098), and Lactiplantybacillus plantarum LB316R (DSM33091).

[0076] In one preferred embodiment, the anti-aging strain is an isolated Lactiplantybacillus plantarum.

[0077] In one preferred embodiment, the anti-aging strain is Lactiplantibacillus plantarum LB244R (DSM32996).

[0078] In one preferred embodiment, the anti-aging strain is a lactic acid bacterium capable of producing at least two bacteriocins, more preferably at least three bacteriocins, and more preferably at least four bacteriocins.

[0079] "Antibacterial metabolites" may include at least one antibacterial lactic acid-producing bacterial metabolite selected from the group including phenyllactic acid, 3-hydroxyphenyllactic acid, 4-hydroxyphenyllactic acid, 3-hydroxypropanaldehyde, 1,2-propanediol, 1,3-propanediol, hydrogen peroxide, ethanol, carbon dioxide, carbonate, propanoic acid, butyric acid, cyclic dipeptides, cyclo(L-Phe-L-Pro), cyclo(L-P-Traps-4-OH-L-Pro), 3-(R)-hydroxydecanoic acid, 3-hydroxy-5-cic dodecanoic acid, 3-(R)-hydroxydodecanoic acid, and 3-(R)-hydroxytetradecanoic acid.

[0080] In one aspect of the present invention, lipids are included in the topical formulation, preferably in the form of natural oils or fats. More preferably, the lipids are derived from vegetable oils or fats. The vegetable oils or fats may be selected from at least one of the following sources: acai, acai berries, sweet almonds, aloe vera, crabwood, apricot kernels, arnica, argan, avocado, babassu, baobab, blackberry seeds, black cumin, blackcurrant seeds, blueberries, borage, Brazil nuts, broccoli seeds, buriti, calendula, camellia seeds, cannabis oil containing CBD and THC, canola, copaiba balsam, Cape chestnut (yango), carrot (dawks carota), castor bean, Chardonnay grapes, caulumugula, cherry kernels, chia seeds, chickweed, coconut, coconut fraction, cottonseed, comfrey, maize, cranbe seeds Cranberry seeds, cucumber seeds, Echium seeds, evening primrose, emu, flaxseed, grape seeds, hazelnuts, hemp seeds, horse chestnut seeds, jojoba, black yonna seeds, kiwi seeds, kukui nuts, macadamia nuts, marula, red hollyhock, manchetti, meadowfoam, milk thistle seeds, moringa, mullein, mustard seeds, neem, olive, palm, papaya seeds, passionflower seeds, peach kernels, peanuts, perilla, pomegranate, Pentaclethra macroloba, pumpkin seeds, raspberry seeds, rice bran, rosehip, St. John's wort oil, safflower, sea buckthorn pulp, shea butter oil, roasted sesame seeds, sesame seeds, soybeans, sunflower, tamanu (Calophyllum) Inophyllum), thistle, tomato, turkey red, Sangre de Drago, walnut, watermelon seed, wheat germ, Abyssinian, rapeseed, beeswax, lanolin, flaxseed, mortierella oil, ongocea, paraffin liquid, pecan, peggy, poppy seed, pracaxi, rapeseed, soybean, toru, tung tree, veronica, wheat germ, yango seed and any combination thereof.

[0081] In preferred embodiments of the present invention, the lipids are derived from vegetable oils, fats, or waxes selected from almond oil, sunflower oil, hemp oil, sweet almond oil, shea butter, cocoa butter, rosehip oil, jojoba oil, golden jojoba oil, rapeseed oil, chamomile oil, calendula oil, evening primrose oil, borage oil, acai oil, sea buckthorn oil, safflower oil, castor oil, olive oil, linseed oil, apricot kernel oil, argan oil, camelina oil, comfrey oil, grape seed oil, kiwi seed oil, malein oil, peach kernel oil, thistle oil, and sesame oil.

[0082] The “therapeutic effective dose” of a compound in the subject matter refers to the amount of the compound in a preparation that, when administered as part of a desired drug regimen (for mammals, preferably humans), alleviates symptoms, improves a condition, or delays the onset of a pathological condition, in accordance with clinically acceptable standards for the disorder or condition being treated or for cosmetic purposes, at a reasonable benefit-risk ratio applicable to any medical treatment.

[0083] As used herein, the terms “to treat” or “to treat” include reversing, reducing, or cessating the symptoms, signs, and underlying pathologies of aging in a manner that improves or stabilizes the condition in question.

[0084] Preferably, changes from baseline may be observed within 20 days, for example, 28 days, 35 days, 46 days, 50 days, or 56 days from the application of the first dose of the composition according to the present invention.

[0085] Herein, and in all descriptions of the range given in the present invention characterized by terms such as "about" or "approximately," it will be apparent to those skilled in the art that the exact numerical range does not need to be indicated by expressions such as "about" or "approx." or "approximately," and instead even slight deviations above or below the indicated number still fall within the scope of the present invention.

[0086] The compositions according to the present invention may preferably comprise a pharmaceutically or cosmetically acceptable vehicle or excipient. In one embodiment of the present invention, the composition may be provided in solid, liquid, viscous, emulsion, or dry form.

[0087] The compositions according to the present invention may preferably be for topical application (topical compositions). Preferably, the compositions may be formulated into emulsions, mists, pastes; talcs; powders; lotions; custards; foams; creams; oils; gels; serums; ointments; sprays or semi-solid formulations.

[0088] The preferred pH of the composition is pH 2.5 to pH 7, more preferably pH 3 to pH 6.5, and even more preferably pH 4 to pH 6.

[0089] A preferred embodiment of the present invention relates to a composition comprising at least one living isolated microorganism for use in the treatment, mitigation, inhibition, prevention, and / or prevention of skin aging.

[0090] A more preferred embodiment of the present invention is a composition comprising at least one live isolated microorganism, (a) A reduction of at least 5% in wrinkles, (b) A reduction of at least 5% in pigmented spot density, and / or (c) relating to compositions for use in treating, mitigating, inhibiting, preventing and / or preventing skin aging by providing an increase in firmness, which is measured as a reduction in suction distance of more than 5%.

[0091] A more preferred embodiment of the present invention relates to a composition for use in the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, comprising at least two live lactic acid bacteria for use in topical treatment of the skin, and providing an increase of at least 5% in dermal density after 28 days of daily topical treatment.

[0092] A further preferred embodiment of the present invention is that the composition comprises at least one live isolated microorganism, (a) a reduction of at least 2% in the thickness of the subepidermal hypoechoic band (SLEB), and / or (b) relating to a composition for use in the treatment, mitigation, inhibition, prevention and / or prevention of skin aging by providing a reduction of at least 2% in TEWL.

[0093] A more preferred embodiment of the present invention is a composition comprising at least one live isolated microorganism, (a) an increase in skin tone of more than 10%, and / or (b) relating to compositions for use in treating, mitigating, inhibiting, preventing and / or preventing skin aging by providing an increase in smoothness of more than 10%.

[0094] A further preferred embodiment of the present invention relates to a method of skin treatment comprising administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days for the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, wherein the method (a) A reduction of at least 5% in wrinkles, (b) A reduction of at least 5% in pigmented spot density, and / or (c) Provides increased tension, which is measured as a reduction in suction distance of more than 5%.

[0095] A more preferred embodiment of the present invention relates to a method of skin treatment comprising administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days for the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, wherein the method (a) an increase of at least 5% in dermal density, (b) an increase in skin moisture content of more than 10%, and / or (c) Provides an increase in skin elasticity of more than 2%.

[0096] Another preferred embodiment of the present invention relates to a method of skin treatment comprising administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days for the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, wherein the method (a) a reduction of at least 2% in the thickness of the subepidermal hypoechoic band (SLEB), and / or (b) Provides at least a 2% reduction in TEWL.

[0097] A further preferred embodiment of the present invention relates to a method of skin treatment comprising administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days for the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, wherein the method (a) an increase in skin tone of more than 10%, and / or (b) Smoothness increase of more than 10%, To provide.

[0098] A further preferred embodiment of the present invention relates to live lactic acid bacteria for use in topical skin treatment, which yields at least a 2% reduction in the thickness of subepidermal hypoechoic bands (SLEBs) after 28 days of daily topical treatment.

[0099] A preferred embodiment of the present invention is a composition for use in the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, comprising at least one live isolated microorganism, administered daily as a topical treatment for 28 days. (a) an increase of at least 5% in dermal density, (b) an increase in skin moisture content of more than 10%, and / or (c) relating to a composition that provides an increase in skin elasticity of more than 2%.

[0100] All patent and non-patent references cited in this application are incorporated herein by reference in their entirety.

[0101] The present invention will be described in more detail in the following non-limiting embodiments. [Examples]

[0102] Example 1. Clinical trial material and method Test product The investigational drug, Probiotic Serum (PS), contains natural plant lipids; shea butter, rapeseed oil, jojoba seed oil, hydrogenated olive oil, hybrid sunflower oil, and live lactiplantibacillus plantarum LB244R (1 x 10 9 It was an oily serum containing CFU / g. Clinical trials

[0103] The study was designed as a single-center trial of topical cream. The characteristics of the study population are shown in Table 1. The study was conducted in accordance with the principles of the Declaration of Helsinki and subsequent amendments, in accordance with the standards for conducting clinical trials of medicinal products, reviewed by an internal review board (Opinion No. 8397 / 2022), and approved by an independent ethics committee for human use. Informed consent was obtained prior to the study.

[0104] [Table 1]

[0105] The 23 participants were enrolled based on being postmenopausal women aged 40–65 years, with normal, mixed / dry to dry skin types, and exhibiting Fitzpatrick's skin phototype II–IV and signs of skin aging. Skin aging was assessed by wrinkles, fine lines, and pigmentation (at least grade 2 to grade 5 on the Bazin scale). Participants were excluded if they had skin marks that interfered with the assessment, a history of known allergies to the product, a history of skin cancer, a change in hormone therapy during the study, prior vitamin A or anti-aging treatment, or planned intensive sun exposure during the study period.

[0106] Participants were instructed to apply PS twice daily and were evaluated three times throughout the study by a dermatologist: at the start of treatment (D0), and after use at 28±2 days and 56±3 days (D28, D56). All evaluations were performed after an acclimatization process of at least 15 minutes in a fully controlled and regulated room (temperature 21±2°C, relative humidity 55±10%). Product suitability and acceptability evaluation

[0107] Suitability was assessed by discomfort reported by the subjects and observable irritation responses determined by skilled dermatologists. Skin ultrasound examination

[0108] Using a Dermascan C ultrasound system (Cortex Technology, Denmark) equipped with an improved 20MHz ultrasound probe, high-density tissue and structure were detected in the dermis of the cheek region. The thickness of the SLEB (subepidermal hypoechoic band) and skin density were calculated. Biomechanical evaluation of the skin

[0109] Skin firmness and elasticity were evaluated using a Cutometer® dual MPA 580, with a 2mm probe applied for 2 seconds using a negative pressure of 450 mbar, followed by a 2-second release. Measurements were obtained in the cheek region, firmness (R0) was interpreted using the suction height (mm), and viscoelasticity was measured by the recovery distance as a percentage of the suction height (R2). Effectiveness evaluation by score

[0110] Dermatologists or qualified technicians assessed pigment spot density scores and crow's feet wrinkle clinical scores according to Bazin (2011) Skin Res Technol May;17(2):135-40.doi:10.1111 / j.1600-0846.2010.00481.x. Similarly, smooth appearance and skin radiance were assessed on a scale of 0 to 9, with 9 being the highest. Evaluation of skin barrier function

[0111] Skin barrier function was measured using transepidermal water loss (TEWL). Measurements in the cheek area of ​​the face were evaluated using Tewameter™ 300 (Courage-Khazaka Electronic GmbH, Cologne, Germany). Evaluation of skin moisture content

[0112] Skin moisture content was indirectly measured by measuring capacitance dependent on the water content of the stratum corneum using a Corneometer CM825 probe connected to a Cutometer Dual MPA 580 (Courage & Khazaka, Germany). Measurements were performed on the cheek area of ​​the face. Measurement values ​​are obtained in arbitrary units as per factory standards.

[0113] statistical analysis Student's t-tests or Wilcoxon signed-rank tests were performed to compare all continuous data between day 0 and days 28 and 56, respectively. A significance level of 95% was adopted. Each subject served as its own criterion to assess relative change.

[0114] result PS tolerance A total of 23 subjects were enrolled in the trial, 21 completed treatment, and 2 dropped out (Table 1). All subjects tolerated treatment with PS, there were no adverse reactions attributable to the product, and the safety of topical application of the cream(s) formulation(s) containing live lactic acid bacteria was highlighted. In essence, the product showed very good dermatological compatibility and tolerability.

[0115] [Table 2]

[0116] Skin ultrasound examination The mean SLEB thickness decreased significantly from 261±69 μm to 245±53 μm (-3.6% decrease, p<0.05) after 28 days of use, and to 237±55 μm (-6.4% decrease, p<0.05) after 56 days. Dermal density was 325±58 pixels / mm² after 56 days.2 From 368±76 pixels / mm 2 This increased. Therefore, PS exerted a favorable anti-aging effect on both SLEB thickness and dermal density in the subjects (Figure 1 and Table 2).

[0117] Biomechanical evaluation of the skin Skin firmness (R0) decreased by 10.8% (p<0.05) at 28 days after application and by 13.1% (p<0.05) at 56 days (Table 2). Furthermore, skin elasticity (R2) increased by 4.70% and 7.40% (p<0.05) at 28 days and 56 days, respectively (Figure 2 and Table 2).

[0118] Clinical scores The probiotic serum had positive effects on crow's feet wrinkles, pigmentation density, smoothness, and skin radiance. Crow's feet wrinkles significantly decreased after 28 days (-17.2%; p>0.05) and 56 days (-22.4%; p>0.05) of use (Table 2). Pigmentation density significantly decreased after 28 days (-16.3%; p>0.05) and 56 days (-27.5%; p>0.05) of use (Table 2). Skin smoothness significantly increased after 28 days (19.4%; p>0.05) and 56 days (33.5%; p>0.05) of use (Table 2). Skin radiance also significantly increased after 28 days of use (26.4%; p>0.05) and after 56 days of use (44.4%; p>0.05) (Table 2).

[0119] Clinically, PS helped reverse the signs of aging skin in the tested area. Ultrasonography results showed a significant decrease in SLEB thickness, which indicates an anti-aging effect as SLEB thickness increases proportionally with age. While SLEB thickness decreased, dermal density increased. Image analysis showed increased dermal density in both the dermal papillary and dermal reticular layers, indicating an increase in collagen bundles, as this is an echogenic marker protein. This suggests a reversal from aging to mature skin and an increase in collagen bundle thickness. This is further emphasized by the biomechanical evaluation of the test, which showed increased skin firmness and elasticity. As the suction distance decreased, skin firmness increased (Table 2, Figure 2A). Firmness is a combination of the elasticity, viscoelasticity, and flexibility of collagen and elastin fibers. This is supported by increased elasticity (Table 2, Figure 2B), suggesting improved strengthening of collagen and elastin fibers. When measured with a 2mm probe, these changes are observed in the stratum corneum and dermal papillary layer.

[0120] Wrinkle formation is associated with sagging and loss of elasticity in the skin, caused by damage to the extracellular matrix of collagen and elastin in the dermis. As the dermis ages, collagen fibers become disordered, resulting in increased wrinkles, roughness, and radiance. Topical treatment with PS significantly reduced crow's feet wrinkles, along with decreased pigmentation density, improved skin radiance, and improved smoothness. These results are a physical manifestation of the ultrasound and biomechanical improvements that underlie the anti-aging benefits of PS (Table 2).

[0121] Furthermore, the increase in smoothness directly reflects the restorative effect of PS on the skin barrier after 56 days of treatment, as demonstrated by TEWL. Following the increase in barrier function, there was also a significant increase in skin moisture content (Table 2).

[0122] In conclusion, 21 participants were enrolled in the PS clinical trial and demonstrated both dermatological suitability and tolerability. PS significantly reduced SLEB thickness, increased dermal density and skin firmness and elasticity, and improved clinical signs of skin aging. The product also improved crow's feet, pigmentation density, and skin radiance. These results suggest that PS is a useful anti-aging product.

[0123] Example 2: Determination of the survival rate of microorganisms applied to the skin. L. plantarum LB316R, L. plantarum LB244R, and L. plantarum LB312R were selected as viable strains and incorporated into topical oily formulations containing almond oil (50% w / w), jojoba oil (49% w / w), and approximately 1% lyophilized microorganisms. The viable cell concentration in each formulation was determined by plate counting on MRS medium.

[0124] Each topical formulation was applied to human skin, and the skin surface was divided into 4x4 cm areas. Each area was wiped once. The number of viable lactic acid bacteria (LABs) was evaluated by wiping with a sterile cotton swab and spreading it on an MRS agar plate. Following the determination of the number of viable LABs, wipes were taken from the 4x4 area every hour, and the survival of LABs on human skin was determined relative to the cell count at the time of application of the topical formulation.

[0125] The number of CFU / g in the topical formulation is approximately 10 per batch. 8 It was determined to be CFU / g. Formulation A (LB316R): 8x10 8 CFU / g Formulation B (LB244R): 7x10 8 CFU / g Formulation C (LB312R): 5x10 8 CFU / g

[0126] During the test, the skin was not covered, exposed to sunlight, rinsed, or washed.

[0127] When applied to human skin, all three strains remain fully viable on the skin for approximately 8-10 hours, after which their survival rate begins to decline.

[0128] Example 3: pigment spot When formulation A of Example 2, containing the L. plantarum LB316R strain, was applied twice daily to photoaged skin, a significant and visible reduction in both pigmented spots and wrinkles was observed after two weeks. Figure 4A shows the skin before treatment, and Figure 4B shows the same area of ​​skin after two weeks of application.

[0129] Skin moisture content, firmness, and smoothness were assessed by the subjects, and all parameters were assessed as having improved after two weeks of treatment.

[0130] Example 4: Placebo-controlled, double-blind clinical trial The second clinical trial was designed as a single-center, double-blind, placebo-controlled vehicle ointment trial. All subjects were randomly assigned to receive either a topical ointment without the active ingredient (placebo ointment) or a topical ointment containing the active ingredient (live Lactobacillus plantarum LB244R).

[0131] Inclusion criteria included: postmenopausal women aged 40–65 years with normal, combination / dry, or dry skin, and visible signs of aging, e.g., fine lines and pigmented spots of the lowest grade 2 on the Bazin scale. Participants were instructed to apply the assigned topical ointment twice daily, morning and evening, to clean skin of the face and around the eyes, and to massage until fully absorbed, for 56 consecutive days.

[0132] All parameters for the subjects were assessed by a skilled technician or dermatologist on days 0, 26, and 56. All assessments were performed after a 15-minute minimal acclimatization process in a room that was fully controlled and regulated to a temperature T=21°C ± 2°C and relative controlled humidity RH=55% ± 10%.

[0133] All subjects included in the study's demographic characteristics were female. [Table 3]

[0134] Skin ultrasound examination Skin ultrasound was measured in the same manner as in the previous study example 1. Tissue density and dermal structure were assessed in the cheek region, randomly assigned to either the left or right side, using a Dermascan C ultrasound system (Cortex Technology, Denmark) with an improved 20 MHz ultrasound probe. The measurements were used to calculate the thickness and skin density of the subepidermal hypoechoic band (SLEB).

[0135] Biomechanical evaluation of the skin The biomechanical parameters of the skin were measured using a Dual-cutometer MPA 580® (Courage & Khazaka Electronic GmbH, Germany) as in Example 1. Measurements were performed using a 2 mm probe, and the skin was mechanically deformed using negative pressure. Negative pressure was generated within the device (450 mBar), and the skin was released after 2 seconds. Penetration depth was determined using a non-optical measurement system. Penetration depth is used as a measure of skin firmness, and the ability of the skin to return to its original position is used as a measure of skin elasticity.

[0136] Skin moisture content Skin moisture content was assessed using a Corneometer CM825 probe connected to a Cutometer dual MPA 580 (Courage & Khazaka, Germany). Measurements were performed in the cheek area of ​​the face, randomly assigned to either the left or right side. Moisture content was assessed by measuring skin capacitance, expressed in arbitrary units (AU) relative to factory standards.

[0137] Visualization of skin structure Skin conditions were visualized and characterized using confocal microscopy with Vivascope® 1500 (Mavig, Germany). The epidermis and reticular dermis were imaged and examined using in vivo confocal laser scanning microscopy (CLSM) or line-field confocal optical coherence tomography (LC-OCT) (Damae, France). Images are generated using an 830 or 800 nm laser striking a defined spot at a specific skin depth. These images can be used to create 3D objects by scanning several optical axis planes and stacking them using suitable microscopy software.

[0138] Clinical evaluation The efficacy of the ointment was evaluated by a dermatologist at baseline and after 28 and 56 days of application. The following parameters were evaluated: Clinical scores for crow's feet wrinkles were assessed using the Bazin scale, clinical scores for blemishes using the Bazin scale, a skin smoothness score on a scale of 0 to 9 (9 meaning very smooth skin, 0 meaning not smooth skin), and a skin luster score on a scale of 0 to 9 (9 meaning very lustrous skin, 0 meaning dull skin).

[0139] These observations were supported by self-assessment questionnaires.

[0140] statistical analysis Statistical analysis was performed on all continuous data comparisons between D0, D28, and D56, as well as between both products, using either Student's t-test or Wilcoxon's signed-rank test. For subjective data, a binomial test was used. A 95% significance level was adopted for all tests.

[0141] For all measured parameters, the active substance performed better than the placebo vehicle ointment, demonstrating a statistically significant effect of topically applied live probiotics. [Table 4] [Brief explanation of the drawing]

[0142] [Figure 1] The ultrasound evaluation shows a decrease in SLEB thickness and an increase in dermal density. [Figure 1A] The bars show the average percentage change in SLEB relative to day 0 of the study. [Figure 1B] The bar graph shows the average percentage change in dermal density relative to day 0 of the study. [Figure 2] This figure shows the improved biomechanical properties of the skin, as measured by a cutometer, with respect to firmness (R0) and elasticity (R2). [Figure 2A] The bar graph shows the average percentage change in skin firmness relative to day 0 of the study. [Figure 2B] The bar graph shows the mean percentage change in skin elasticity relative to day 0. The significance level compared to baseline values ​​was tested using the Wilcoxon signed-rank test for paired data. * indicates p<0.05. [Figure 3] This figure shows the clinical evaluation of crow's feet wrinkles, pigment spot density, skin tone, and smoothness. [Figure 3A] The bar shows the average percentage change in crow's feet wrinkles, with the subject's baseline at day 0. [Figure 3B] The bar graph shows the average percentage change in pigment spot density relative to day 0 of the study. [Figure 3C] The bar shows the average percentage change in skin tone, relative to day 0 of the subject's condition. [Figure 3D] The bars represent the mean percentage change in smoothness relative to day 0 of the study. Significance levels compared to baseline values ​​were tested using the Wilcoxon signed-rank test for the paired data. * indicates p<0.05. [Figure 4A] Visual evaluation of wrinkles and hyperpigmentation before treatment. [Figure 4B] Visual evaluation of wrinkles and hyperpigmentation after 2 weeks of treatment with twice-daily application.

Claims

1. A composition comprising at least one live isolated microorganism for use in the treatment, mitigation, inhibition, prevention, and / or prevention of skin aging.

2. The composition comprises at least one living isolated microorganism, (a) A reduction of at least 5% in wrinkles, (b) A reduction of at least 5% in pigmented spot density, and / or (c) The composition according to claim 1 for use in treating, mitigating, inhibiting, preventing and / or preventing skin aging by providing an increase in firmness, which is measured as a reduction in suction distance of more than 5%.

3. The composition according to claim 1, for use in treating, mitigating, inhibiting, preventing and / or preventing skin aging, wherein the composition comprises at least two live lactic acid bacteria for use in topical treatment of the skin, and after 28 days of daily topical treatment, an increase of at least 5% in dermal density is obtained.

4. The composition comprises at least one living isolated microorganism, (a) a reduction of at least 2% in the thickness of the subcutaneous hypoechoic band (SLEB), and / or (b) The composition according to claim 1 for use in treating, mitigating, inhibiting, preventing and / or preventing skin aging by providing a reduction of at least 2% in TEWL.

5. The composition comprises at least one living isolated microorganism, (a) an increase in skin tone of more than 10%, and / or (b) The composition according to claim 1 for use in treating, mitigating, inhibiting, preventing and / or preventing skin aging by providing an increase in smoothness of more than 10%.

6. The composition according to any one of claims 1 to 5, wherein the microorganism is a lactic acid bacterium.

7. The composition according to claim 6, wherein the lactic acid bacteria produce antimicrobial metabolites and / or bacteriocins.

8. The composition according to any one of claims 1 to 7, wherein the change from baseline is observed within 20 days, for example, within 28 days, for example, within 35 days, for example, within 46 days, for example, within 50 days, for example, within 56 days, from the application of a first dose of the composition.

9. A localized composition according to any one of claims 1 to 8.

10. The composition according to any one of claims 1 to 9, wherein the topical composition is a cream, lotion, spray, solution, gel, serum, ointment, fat, oil, powder, paste, foam, plaster, paint, adhesive, suspension, or emulsion.

11. The composition, for example, the topical composition, contains at least 10 ４ CFU / g, for example, at least 10 ５ CFU / g, for example, at least 10 ６ CFU / g, for example, at least 10 ７ CFU / g, for example, at least 10 ８ The composition according to any one of claims 1 to 10, comprising live microorganisms at a concentration of CFU / g.

12. The composition according to any one of claims 1 to 11, further comprising at least one natural lipid.

13. A method of skin treatment comprising administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days for the treatment, mitigation, inhibition, prevention and / or prevention of skin aging.

14. For the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, the treatment includes administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days. (a) A reduction of at least 5% in wrinkles, (b) A reduction of at least 5% in pigmented spot density, and / or (c) A method of skin treatment according to claim 13, which provides an increase in firmness, measured as a reduction in suction distance of more than 5%.

15. For the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, the treatment includes administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days. (a) an increase of at least 5% in dermal density, (b) an increase in skin moisture content of more than 10%, and / or (c) Increase in skin elasticity of more than 2%, A method for treating skin according to claim 13, which provides the following.

16. For the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, the treatment includes administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days. (a) a reduction of at least 2% in the thickness of the subcutaneous hypoechoic band (SLEB), and / or (b) A decrease of at least 2% in TEWL, A method for treating skin according to claim 13, which provides the following.

17. For the treatment, mitigation, inhibition, prevention and / or prevention of skin aging, the treatment includes administering a live topical composition containing at least one isolated lactic acid bacterium for at least 28 days. (a) an increase in skin tone of more than 10%, and / or (b) Smoothness increase of more than 10%, A method for treating skin according to claim 13, which provides the following.

18. The living microorganism is at least 10 ４ CFU / cm ２ , for example at least 10 ５ CFU / cm ２ , for example at least 10 ６ CFU / cm ２ , for example at least 10 ７ CFU / cm ２ , for example at least 10 ８ CFU / cm ２ The method according to claims 13 to 17, which is applied to the skin at a concentration of.

19. The method according to any one of claims 13 to 18, wherein the live microorganism is applied to the skin at least once a day.

20. The method according to any one of claims 13 to 19, wherein the live microorganisms are able to survive on the skin for at least 8 hours after topical application.

21. Live lactic acid bacteria for use in topical skin treatment, resulting in at least a 2% reduction in the thickness of subepidermal hypoechoic bands (SLEBs) after 28 days of daily topical treatment.

22. Live lactic acid bacteria for use in topical skin treatment, after 28 days of daily topical treatment, (a) an increase of at least 5% in dermal density, and / or (b) an increase in skin moisture content of more than 10%, and / or (c) Live lactic acid bacteria that provide a more than 2% increase in skin elasticity.

23. The lactic acid bacteria described above were isolated, and at least 10 ４ CFU / cm ２ For example, at least 10 ５ CFU / cm ２ For example, at least 10 ６ CFU / cm ２ For example, at least 10 ７ CFU / cm ２ For example, at least 10 ８ CFU / cm ２ Live lactic acid bacteria according to claim 19 or 20, applied to the skin at the specified concentration.

24. The live lactic acid bacteria according to claims 19 to 21, wherein the lactic acid bacteria belong to the Lactobacillus family.

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