Probiotic Compositions for the Treatment of Acne - Patent application

Abstract

The present invention provides a probiotic composition comprising the bacterial strain L. rhamnosus CECT 30031 and the bacterial strain Arthrospira platensis (Spirulina paraca). The disclosed probiotic composition is useful for treating acne.

Description

[Technical Field]

[0001] Probiotic compositions useful for the treatment of acne are provided. [Background technology]

[0002] The term probiotics is defined as "live microorganisms that, when consumed in sufficient amounts, confer a health benefit on the host" (World Health Organization and Food and Agriculture Organization of the United Nations, 2001). The earliest report on probiotics dates back to 1907, when Elie Metchnikoff described a correlation between the consumption of lactic acid-producing bacteria in yogurt and longevity. In the past few decades, there has been renewed interest in probiotics, not only with regard to digestive health, but also in the management of inflammatory diseases.

[0003] Oral probiotics have been shown to improve insulin sensitivity in animal models and regulate the release of proinflammatory cytokines in the skin through interactions with gut-associated lymphoid tissue (NPL 1). Indeed, the gut-brain-skin axis suggests a mechanism linking gastrointestinal health, influenced by interactions with oral probiotics, with skin health and integrity (NPL 2). In this context, some strains of Lactobacillus have been shown to have systemic anti-inflammatory effects. Studies have shown that Lactobacillus reuteri induces systemic anti-inflammatory cytokines, such as interleukin (IL)-10. Soluble factors derived from L. reuteri inhibit the production of proinflammatory cytokines, and culture supernatants of mouse-derived L. reuteri 6798 inhibit tumor necrosis factor (TNF) production by activated macrophages (NPL 3).

[0004] Several strains of Lactobacillus also exhibit anti-inflammatory properties. The addition of Lactobacillus paracasei NCC2461 has been shown to inhibit neurogenic inflammation in a skin model, and the addition of L. paracasei NCC2461 to lymphocyte cultures has been shown to strongly inhibit CD4+ T cell proliferation in a dose-dependent manner and induce the anti-inflammatory cytokines IL-10 and TGF-β. Mice receiving probiotics for 7 days exhibited significantly enhanced antibody responses and in vivo T cell-mediated immune responses, indicating that L. paracasei influences both B cell and T cell function. Similarly, mice treated with Lactobacillus casei exhibited enhanced IL-10 production and enhanced ability to promote regulatory T cell function. The increase in regulatory T cells suggests that this probiotic may help balance the immune system's response to stimuli (Non-Patent Document 1). As a result, certain probiotic strains can potentially boost appropriate immune responses to, for example, harmful pathogenic threats, while suppressing unnecessary immune responses seen in chronic inflammatory conditions.

[0005] Disruption of skin barrier function is a known side effect of many acne medications, including topical retinoids and benzoyl peroxide. The inflammation, irritation, and dryness caused by these medications can lead to acne therapies. This may adversely affect compliance with treatment. Rosacea and atopic dermatitis are other skin conditions in which the skin barrier is impaired, and strengthening the skin barrier improves symptoms. Oral ingestion of certain probiotic strains has been shown to improve skin barrier function and affect skin moisturization and transepidermal water loss. In fact, Non-Patent Document 4 tested the effects of oral supplementation with L. paracasei NCC2461 versus placebo in healthy female volunteers in a randomized, placebo-controlled clinical trial. Skin sensitivity was monitored using the capsaicin test, and skin barrier function was measured using transepidermal water loss and dermatological assessment. Both skin sensitivity and skin barrier function improved in the probiotic group. Furthermore, the probiotic group showed elevated serum levels of TGF-β after 29 days, whereas the placebo group did not. TGF-β has been shown to play an important role in skin integrity.

[0006] Therefore, overall, probiotics regulate the development of the immune system, often shifting the immune response to a regulatory and anti-inflammatory state. This ability of probiotics to alter chronic inflammatory conditions suggests that probiotics may have a role in the treatment of chronic inflammatory conditions ranging from inflammatory bowel disease to reactive airway disease, acne, rosacea, atopic dermatitis, and photoaging (Non-Patent Document 5). In this regard, the evidence for probiotic intervention in acne in human studies is summarized in Table 1 below.

[0007] [Table 1] [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Hacini-Rachinel G, Gheit H, Le Luduec JB, Dif F, Nancey S, Kaiserlian D. Oral probiotic control of skin inflammation by acting on both effector and regulatory T cells. PLoS One. 2009;4:e4903

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[0009] However, there remains a need to provide further probiotic compositions that result in significant improvement in the overall treatment of acne, particularly according to known and accepted scoring systems such as the Global Acne Grading System (GAGS) or AGSS (Acne Global Severity Scale). [Brief explanation of the drawings]

[0010] [Figure 1] FIG. 1 is a consort diagram used in the probiotic blend study of Example 1. [Figure 2] FIG. 1 shows the GAGS (Global Acne Scoring System) index assessment at 6 and 12 weeks for the probiotic blend study of Example 1. [Figure 3] FIG. 1 shows the AGSS (Acne Global Severity Scale) index assessment at 6 and 12 weeks for the probiotic blend study of Example 1. [Figure 4] FIG. 1 shows the assessment of the number of inflamed acne lesions at 6 and 12 weeks for the probiotic blend study of Example 1. [Figure 5] FIG. 1 shows patient opinion ratings and subjective ratings at 6 and 12 weeks for the probiotic blend study of Example 1. DETAILED DESCRIPTION OF THE INVENTION

[0011] In the present invention, the inventors have determined that the amount of cellulose in the cellulose is at least 1×10 per gram. 9 A novel maltodextrin-based formulation, designated Bths-03, containing a mixture of two strains containing colony-forming units (cfu) was tested (see Example 1). Specifically, the mixture of two strains included the bacterial strains Lactobacillus rhamnosus CECT 30031 and Arthrospira platensis BEA_IDA_0074B (also known as Spirulina paraca (S. paraca)).

[0012] L. rhamnosus is a bacterium commonly used as a probiotic and is found primarily in yogurt and other dairy products, including infant formula. The scientific classification of L. rhamnosus is: Domain: Bacteria, Phylum: Firmicutes, Class: Bacillus, Order: Lactobacillus, Family: Lactobacillaceae, Genus: Lactobacillus, Species: Lactobacillus rhamnosus.

[0013] Arthrospira platensis is an aquatic, prokaryotic, filamentous, Gram-negative bacterium in the domain Bacteria; phylum Cyanobacteria; class Blue-green algae; genus Cyanobacteria. It is often classified as a blue / green microalga, despite being a prokaryotic bacterial microorganism. Its commercialized biomass is commonly known as "spirulina," and its production makes it the most widely cultivated microorganism in the world. Arthrospira platensis is therefore also known as Spirulina paraca (S. paraca).

[0014] Using the above mixture, the inventors conducted a pilot study involving patients with moderate acne vulgaris who had been treated with topical and systemic antibiotics, as shown in Example 1 attached herein. As shown in Example 1, the response rate according to the Global Acne Scoring System (GAGS) was 61.8% in the probiotic group and 35.4% in the placebo group. This absolute difference of approximately 26 points in improvement rate was considered clinically relevant and statistically significant. Another clinically important variable measured was the Acne Global Severity Scale (AGSS) variable. Differences were also observed between the groups, particularly a 58% improvement in the index in the probiotic group and a 33% improvement in the placebo group. Regarding the variable "number of inflammatory lesions," this variable showed clinically and statistically relevant differences between the two groups, with 76.5% of lesions improving in the probiotic group and only 23.78% of lesions improving in the placebo group. This last-mentioned variable was the one that showed the greatest improvement during treatment, favoring the group treated with the probiotic blend. Additionally, the patient's subjective impression showed a 29.58% improvement at the end of treatment in the probiotic group, compared with a 4.75% improvement in the placebo group. Furthermore, all main variables (primary and secondary) showed more favorable scores in the probiotic group when comparing data with the placebo group, some of which had important clinical relevance and statistical significance. Adherence to treatment was 90% in both groups, and the number of side effects described was low and not attributable to the treatment used in the study.

[0015] Based on these results, the inventors can conclude that the present invention provides a new and improved treatment of acne with a probiotic composition comprising a combination of at least two strains, in particular a bacterial strain belonging to the species L. rhamnosus and a bacterial strain belonging to the species Arthrospira platensis (Spirulina paraca). More specifically, the present invention relates to at least the bacterial strain Lactobacillus rhamnosus (hereinafter referred to throughout the specification as L. rhamnosus CECT 30031), which was deposited on December 3, 2019 by Bioithas (03080 Sant Vicent del Raspeig, Alicante, Spain) at the Spanish Type culture Collection (CECT) (C / Catedratico Agustin Escardino, 9. 46980 Paterna (Valencia), Spain) under the accession number CECT 30031, designated AcnePro-Bths, and the bacterial strain Lactobacillus rhamnosus (hereinafter referred to throughout the specification as L. rhamnosus CECT 30031), which was deposited on December 3, 2019 by Bioithas (03080 Sant Vicent del Raspeig, Alicante, Spain) at the Spanish Type culture Collection (CECT) (C / Catedratico Agustin Escardino, 9. 46980 Paterna (Valencia), Spain) under the accession number CECT 30031, designated SPF-bioithas 001. This invention relates to a combination of the Arthrospira platensis strain deposited at the National Institute for Microbiology and Biology (NIH) in Spain on October 25, 2019 under accession number BEA_IDA_0074B (hereinafter referred to throughout this specification as S. paraca BEA_IDA_0074B or Arthrospira platensis BEA_IDA_0074B).

[0016] Furthermore, as shown in the accompanying Examples, prior to the studies or trials described in Example 1, the strains L. rhamnosus CECT 30031 and S. paraca BEA_IDA_0074B were each evaluated separately for the treatment of acne vulgaris. To this end, a small pilot study was conducted with five patients in each of three branches of the study using L. rhamnosus CECT 30031 alone, S. paraca BEA_IDA_0074B alone, or CECT 30031 and S. paraca BEA_IDA_0074B. The results of this study are presented in Example 2 and reflect how the combined use of both strains significantly enhanced and thus improved efficacy compared to the effects of each strain used separately.

[0017] Furthermore, prior to the testing also described in Example 1, a probiotic composition containing the S. paraca BEA_IDA_0074B strain was compared with a probiotic composition containing a known strain (strain 1. L. rhamnosus SP1) as disclosed in "Fabbrocini G, Bertona M, Picazo O, Pareja-Galeano H, Monfrecola G, Emanuele E. Supplementation with Lactobacillus rhamnosus SP1 normalizes skin expression of genes implicated in insulin signaling and improves adult acne. Benef Microbes. 2016; 7 (5): 625-630. doi: 10.3920 / BM2016.0089." Such known strains have been considered effective in the prior art for the treatment of acne vulgaris (see Table 1 above). The results are shown in Example 3.

[0018] Thus, the results presented in Example 3 for L. rhamnosus strain SP1 and the data presented in Example 2 for L. rhamnosus CECT 30031 (both when used individually) demonstrate that L. rhamnosus strain CECT 30031 is significantly more effective in treating acne vulgaris than L. rhamnosus strain SP1. Furthermore, based on the results presented in Examples 1-3, it is also evident that the anti-acne effect is significantly enhanced when L. rhamnosus CECT 30031 is used in combination with S. paraca BEA_IDA_0074B.

[0019] Thus, a first aspect of the present invention relates to a probiotic composition comprising a combination of the strains L. rhamnosus CECT 30031 and S. paraca BEA_IDA_0074B.

[0020] In the present invention, the term "probiotic composition", when used in any of the above aspects, is understood as a composition comprising at least one microorganism which, when ingested, interacts with the metabolism of an individual and produces a beneficial effect in the individual. In the present invention, the probiotic composition thus comprises at least a combination of the microorganisms L. rhamnosus CECT 30031 and S. paraca BEA_IDA_0074B.

[0021] A second aspect of the present invention relates to a probiotic composition according to the first aspect of the invention for use in the treatment and / or prevention of acne (particularly a method for alleviating, reducing, treating and / or preventing acne), particularly the treatment and / or prevention of acne vulgaris, and more particularly the prevention or treatment of comedones (e.g. reducing the number of comedones and / or preventing the worsening of comedones) in a subject or individual in need of such treatment and / or prevention.

[0022] A third aspect of the present invention relates to a probiotic composition comprising the Arthrospira platensis (S. paraca) strain deposited at Banco Espanol de Algas under accession number BEA_IDA_0074B for use in a method for alleviating, reducing, treating and / or preventing acne, particularly acne vulgaris, and more particularly comedones, in a subject, preferably a human subject, in need thereof.

[0023] A fourth aspect of the present invention relates to a probiotic composition comprising the Lactobacillus rhamnosus strain deposited under the Budapest Treaty at the Spanish Type culture Collection (CECT) under accession number CECT 30031 for use in a method for alleviating, reducing, treating and / or preventing acne, in particular acne vulgaris, and more particularly comedones, in a subject, preferably a human subject, in need thereof.

[0024] In preferred embodiments of the second, third and fourth aspects of the present invention, the acne is selected from the group consisting of acne vulgaris, acne rosacea, acne conglobata, acne fulminans, gram-negative folliculitis and facial pyoderma, preferably the acne is acne vulgaris.

[0025] In another preferred embodiment of the second, third and fourth aspects of the present invention, the probiotic composition is administered orally to the subject.

[0026] Another aspect of the present invention relates to compositions herein provided for use in the treatment and / or prevention of disorders characterized by excessive sebum secretion, particularly disorders that result in comedones and blockage of the pilosebaceous unit. Another aspect relates to compositions herein provided for use in cleansing the skin, particularly in the case of oily or acne-prone skin.

[0027] In the present invention, the term "subject" is equivalent to the term "individual," and therefore, both terms can be used interchangeably herein. "Subject" refers to any animal of any species, as well as any individual. Examples of subjects include, but are not limited to, animals of commercial interest, such as birds (hens, ostriches, chicks, geese, partridges, etc.), rabbits, hares, pet animals (dogs, cats, etc.), sheep, goat cattle (goats, etc.), Sus scrofa (boars, pigs, etc.), equine livestock (horses, ponies, etc.), bovine animals (bulls, cows, steers, etc.); animals of hunting interest, such as bucks, deer, reindeer, etc.; and humans. However, in certain embodiments, the subject is a mammal, and in particular, the mammal is a human of any race, sex, or age.

[0028] In the present invention, the term "prevention" means avoiding the occurrence of a disease or pathological condition in an individual, particularly when the individual has a predisposition to the pathological condition but has not yet been diagnosed. In a preferred embodiment of the present invention, the disease or pathological condition is "acne."

[0029] In the present invention, the term "treating" or "treatment" includes inhibiting the disease or pathological condition, i.e., arresting its progression; alleviating the disease or pathological condition, i.e., causing regression of the disease or pathological condition; alleviating or reducing at least one symptom or sign of the disease or pathological condition; and / or stabilizing the disease or pathological condition in an individual. In a preferred embodiment of the present invention, the disease or pathological condition is "acne."

[0030] It is noted herein that in the context of the present invention, the term "acne" is understood as a disease of the sebaceous follicles, often called pores. At the base of each hair follicle is a gland called a sebaceous gland that produces sebum. Sebum is an oily substance that keeps the skin moist and supple and, under normal circumstances, travels along the hair follicle to the skin's surface. A blemish begins approximately two to three weeks before it appears at the skin's surface. As the skin regenerates, old cells die and are shed. When cells are unevenly shed and clump together with sebum, they form a plug. The sebum that normally drains to the surface becomes clogged, and bacteria begin to grow. The rapid proliferation of bacteria and the accumulation of sebum cause the hair follicle to enlarge, resulting in a mild form of acne called a non-inflammatory comedone. Both whiteheads and blackheads begin as "microcomedones" that then develop into skin defects called comedones, either whiteheads or blackheads. Acne is the growth of trapped sebum and bacteria (Propionibacterium acnes) within blocked hair follicles. Sebaceous glands are most abundant on the face, chest, back, neck, and scalp; as a result, these are the most common areas for acne. The most common factors that cause acne are hormones, increased sebum production, bacteria (Propionibacterium acnes), systemic and local inflammation, and changes within the hair follicle. Acne can progress to red, inflammatory acne lesions called papules, pustules, and nodules.

[0031] As used herein, the term acne is not limited to any particular type, as there are many types of acne ranging in severity from mild to extremely disfiguring. In particular, the term "acne" as used herein refers to any type of acne, particularly those selected from the group consisting of acne vulgaris, acne rosacea, acne conglobata, acne fulminans, gram-negative folliculitis, and facial pyoderma. In particular, the type of acne is acne vulgaris.

[0032] Acne vulgaris is the most common form of acne, including several types of pimples. These acne lesions include blackheads, whiteheads, papules, pustules, nodules, and cysts. Mild to moderate acne vulgaris is characterized by whiteheads, blackheads, papules, and pustules. Whiteheads form when a pore becomes completely blocked, trapping sebum, bacteria, and dead skin cells beneath the skin's surface, resulting in a white appearance. Whiteheads typically have a faster life cycle than blackheads. Blackheads form when a pore becomes only partially blocked, slowly releasing some of the trapped sebum, bacteria, and dead skin cells to the surface. Their black color is due to the reaction of melanin, a pigment found in the skin, with oxygen in the air. Blackheads tend to be stable structures. Blackheads often take a long time to remove because their contents are released to the surface very slowly. Papules are small, red, soft bumps without a tip. Papules are the earliest stage in the development of what is usually considered a typical "pimple." Papules are intermediate between the non-inflammatory and inflammatory stages in the progression of acne. Pustules are similar to whiteheads but are inflamed and appear as red rings with white or yellow centers.

[0033] Severe acne vulgaris is characterized by nodules and cysts. Nodular acne consists of acne spots that can be much larger, very painful, and can last for months. Nodules are large, hard lumps below the surface of the skin. Scarring is common with nodules. Acne cysts can be similar in appearance to nodules, but are pus-filled and can be described as having a diameter of 5 mm or more. Cysts can be painful, and scarring is common with cystic acne.

[0034] Acne rosacea can be similar in appearance to acne vulgaris, described above, and the two types of acne are sometimes confused. Rosacea affects millions of people, most of whom are over the age of 30. Rosacea manifests as a red rash usually localized to the cheeks, nose, forehead, and chin. The redness is often accompanied by bumps, pimples, and skin imperfections. Blood vessels may also become more visible on the skin. Blackheads are not included in rosacea. Rosacea is more common in women, but is often more severe in men. If left untreated, it can cause swelling and excess tissue growth in the nose, a condition called rhinophyma.

[0035] Acne conglobata is the most severe form of acne vulgaris and is more common in men. It is characterized by numerous large lesions that may connect to each other and be accompanied by widespread blackheads. Acne conglobata can cause significant and irreversible damage to the skin, resulting in disfiguring scars. Acne conglobata is found on the face, chest, back, buttocks, upper arms, and thighs. The age of onset of acne conglobata is usually between 18 and 30 years of age, and the condition can remain active for many years.

[0036] Acne fulminans is a sudden onset of acne conglobata that usually affects young men. Symptoms of severe nodulocystic, often ulcerative acne are evident. As with acne conglobata, extremely disfiguring scars are common. Acne fulminans is unique in that it is also accompanied by fever and joint pain.

[0037] Gram-negative folliculitis is a bacterial infection characterized by pustules and cysts that can arise as a complication of long-term antibiotic treatment for acne vulgaris. Gram-negative folliculitis is a rare condition, and its prevalence in men versus women is unknown.

[0038] Pyoderma facialis is a severe form of facial acne that typically affects only women between the ages of 20 and 40 and is characterized by large, painful nodules, pustules, and ulcers that can leave scars. Pyoderma facialis can develop suddenly and may occur on skin in women who previously had no acne. Pyoderma facialis is limited to the face and usually does not last more than a year, although it can cause skin irritation in a very short period of time.

[0039] It is noted that the present invention also contemplates a microorganism or bacterium derived from the microorganism L. rhamnosus CECT 30031 that has a 16s rRNA sequence that is at least 99% identical to the 16s rRNA sequence of the L. rhamnosus strain of SEQ ID NO: 1 of L. rhamnosus CECT 30031 and that retains the ability of said L. rhamnosus CECT 30031 to reduce and / or ameliorate the progression of acne in an individual in need thereof.

[0040] The present invention also contemplates a microorganism or bacterium derived from the microorganism S. paraca BEA_IDA_0074B (Arthrospira platensis BEA_IDA_0074B) that has a 16s rRNA sequence that is at least 95% identical to the 16s rRNA sequence of S. paraca BEA_IDA_0074B and that retains the ability of S. paraca BEA_IDA_0074B to reduce and / or ameliorate acne progression in an individual in need thereof.

[0041] Such derived bacteria may be part of, or may completely or partially replace, any of the microorganisms L. rhamnosus CECT 30031 and / or S. paraca BEA_IDA_0074B in any of the above probiotic compositions of the invention, so long as they retain the ability to reduce and / or ameliorate the progression of acne in individuals in need thereof.

[0042] In the context of the present invention, "retaining the ability to reduce and / or ameliorate acne progression in an individual in need thereof" is to be understood as a significant improvement in the GAGS (Global Acne Grade Score) index of at least 26% compared to placebo-treated subjects, as well as an improvement of at least two categories in the AGSS (Acne Global Severity Scale) index of 58% in the probiotic group compared to 33% in subjects in the placebo group, optionally an improvement of at least 51% in the number of inflamed lesions compared to the results obtained by treating subjects suffering from acne with oral antibiotics and / or topical treatment with salicylic acid, azelaic acid and / or dapsone.

[0043] Examples of strains or microorganisms derived from strains included in any of the probiotic compositions of the present invention described above may be mutants and genetically modified organisms that exhibit variations in their genomes compared to the genomes of the strains of the present invention from which they are derived, but that do not affect the ability of the strains to reduce and / or ameliorate acne progression in individuals. Strains derived from L. rhamnosus CECT 30031 and S. paraca BEA_IDA_0074B can be created naturally or intentionally by mutagenesis methods known in the art, such as, but not limited to, growing the parent strain in the presence of a mutagen or stressor, or by genetic engineering performed to modify, delete, and / or insert specific genes. Thus, as indicated above, the present invention also contemplates genetically modified organisms derived from L. rhamnosus CECT 30031 and S. paraca BEA_IDA_0074B that retain the ability to reduce and / or ameliorate acne progression in individuals and, therefore, be used to treat acne. Certain derivative strains have a 16s rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to the 16s rRNA sequence (SEQ ID NO: 1) of the parent strain and retain the ability of the parent strain to reduce and / or ameliorate the progression of acne in individuals in need thereof.

[0044] SEQ ID NO:1:

[0045] Furthermore, the present invention contemplates cellular components, metabolites, and molecules secreted by L. rhamnosus CECT 30031 and S. paraca BEA_IDA_0074B strains, as well as compositions containing the above components and their use for the treatment and / or prevention of acne. Bacterial cellular components may include cell wall components (such as, but not limited to, peptidoglycan), nucleic acids, membrane components, and the like, such as proteins, lipids, and carbohydrates, and combinations thereof (such as lipoproteins, glycolipids, or glycoproteins). Metabolites include any molecules produced or modified by bacteria or microalgae as a result of metabolic activity during growth, use in technological processes, or storage of the product (the first probiotic composition of the present invention). Examples of these metabolites include, but are not limited to, organic and inorganic acids, proteins, peptides, amino acids, enzymes, lipids, carbohydrates, lipoproteins, glycolipids, glycoproteins, vitamins, salts, minerals, and nucleic acids. Secreted molecules include any molecules secreted or released externally by bacteria during growth, use in technological processes (e.g., food processing or pharmaceuticals), or product storage. Examples of these molecules include, but are not limited to, organic and inorganic acids, proteins, peptides, amino acids, enzymes, lipids, carbohydrates, lipoproteins, glycolipids, glycoproteins, vitamins, salts, minerals, or nucleic acids.

[0046] As will be understood by those skilled in the art, any of the probiotic compositions of the present invention may be formulated for pharmaceutical administration, i.e., form part of a medicinal product administered to a subject (either orally, topically, etc., preferably orally), and / or formulated for food administration, i.e., form part of a food ingested in the subject's diet, and thus may be administered orally. Thus, in certain embodiments, any of the compositions or probiotic compositions of the present invention may be a pharmaceutical composition (hereinafter, "pharmaceutical composition of the present invention") and / or a nutritional composition (hereinafter, "nutritional composition of the present invention"). Such pharmaceutical compositions of the present invention may therefore be used for the treatment and / or prevention of diseases characterized by excessive sebum secretion, particularly diseases that block the pilosebaceous unit, resulting in comedones. In particular, such pharmaceutical compositions of the present invention are used for the prevention or treatment of acne, particularly acne vulgaris, and more specifically, for the prevention or treatment of comedones. Comedones are small, skin-colored bumps (papules) frequently found on the forehead and chin of people with acne. A single lesion is a comedo. Open comedones are blackheads, which are black not because of dirt but because of surface pigment (melanin), while closed comedones are whiteheads, which are completely blocked hair follicles.

[0047] The pharmaceutical compositions of the present invention therefore comprise the microorganisms L. rhamnosus CECT 30031 and / or S. paraca BEA_IDA_0074B (or strains derived therefrom as defined above) in any concentration and may additionally comprise one or more ingredients or compounds with any biologically, pharmacologically, and / or veterinarily useful activity in the prevention and / or treatment of acne, in particular for the treatment or prevention of comedones. Such pharmaceutical compositions may further comprise one or more ingredients capable of further increasing, enhancing, and / or promoting the activity of the strains comprised in any of the probiotic compositions of the present invention upon administration to a subject. As will be understood by those skilled in the art, the additional ingredients or compounds must be compatible with any of the strains in the probiotic compositions of the present invention. In the context of the present invention, the term "pharmaceutical composition" also encompasses veterinary compositions.

[0048] In certain embodiments, the pharmaceutical compositions of the present invention further comprise a pharmaceutically acceptable carrier and / or excipient.

[0049] The term "excipient" refers to a substance that aids in the preparation of a pharmaceutical composition in the sense of aiding the absorption of any component or compound of any of the probiotic compositions of the present invention, i.e., the strains of the present invention, or stabilizing the component or compound, and / or providing consistency or flavor for a more pleasant taste. Thus, excipients may have, by way of example and not limitation, the function of binding components (e.g., starch, sugar, or cellulose), sweetening, coloring, protecting the active ingredient (e.g., isolating the active ingredient from air and / or moisture), filling a pill, capsule, or any other presentation, or disintegrating to facilitate the dissolution of the ingredient, but this does not exclude other excipients not mentioned in this paragraph. The term "excipient" is therefore defined as a material included in a galenic form and added to an active ingredient or its conjugates to enable their preparation and stability, modify their organoleptic properties, or determine the physicochemical properties and bioavailability of a pharmaceutical composition. A "pharmaceutically acceptable" excipient must enable the activity of the component or compound of the pharmaceutical composition, i.e., be compatible with the strains of the present invention.

[0050] A "galenical form" or "pharmaceutical form" is a configuration in which the active ingredient and excipients are adapted to provide a pharmaceutical composition or drug of the invention. It is defined by the combination of the form in which the pharmaceutical composition is provided by the manufacturer and the form in which it is to be administered.

[0051] A "vehicle" or "carrier" is a particularly inert substance. The function of a carrier is to facilitate the incorporation of other ingredients or compounds, allow for better dosing and administration, and / or provide consistency and form to the pharmaceutical composition. Thus, a carrier is a substance used to dilute any of the ingredients or compounds of the pharmaceutical composition of the present invention to a given volume or weight, or a substance that can allow for better dosing and administration and / or provide consistency and form to the drug without diluting these ingredients or compounds. When the dosage form is liquid, a pharmaceutically acceptable carrier is a diluent. Carriers can be natural or non-natural. Examples of pharmaceutically acceptable carriers include, but are not limited to, water, saline, alcohol, vegetable oils, polyethylene glycol, gelatin, lactose, starch, amylose, magnesium stearate, talc, surfactants, silicic acid, viscous paraffin, flavor oils, mono- and diglycerides of fatty acids, fatty acid esters petroetrals, hydroxymethylcellulose, polyvinylpyrrolidone, etc.

[0052] Furthermore, excipients and carriers must be pharmacologically acceptable, that is, they are tolerated and evaluated so as not to cause harm to the subject to whom they are administered.

[0053] In any case, the dosage form of the pharmaceutical composition is adapted to the type of administration used.Therefore, the composition can be given in the form of a solution or any other clinically acceptable dosage form and in a therapeutically effective amount.The pharmaceutical composition can therefore be formulated into a solid, semi-solid or liquid preparation, such as a tablet, capsule, powder (e.g., obtained by lyophilization (freeze-drying) or air-drying), granules, solution, suppository, gel or microsphere.In certain embodiments, the pharmaceutical composition is formulated for administration in liquid or solid form.In one embodiment, the composition is in the form of a gelatin capsule.

[0054] In another particular embodiment, the solid formulation is selected from the group consisting of tablets, lozenges, confectioneries, chewable tablets, chewing gum, capsules, sachets, powders, granules, coated particles or coated tablets, tablets, pills, troches, gastro-resistant tablets and capsules, and dispersible strips and films.

[0055] In another particular embodiment, the liquid formulation is selected from the group consisting of oral solutions, suspensions, emulsions and syrups.

[0056] Similarly, various systems are known that can be used for sustained-release administration of any of the probiotic, pharmaceutical, or nutritional compositions of the present invention, including, for example, encapsulation in liposomes, microbubbles, microparticles, or microcapsules. Suitable sustained-release forms, as well as materials and methods for their preparation, are well known in the state of the art. Thus, any orally administrable form of the probiotic composition of the present invention is a sustained-release form that further comprises at least one coating or matrix. The sustained-release coating or matrix includes, but is not limited to, natural, semi-synthetic, or synthetic polymers, water-insoluble or modified waxes, fats, fatty alcohols, fatty acids, natural, semi-synthetic, or synthetic plasticizers, or a combination of two or more thereof. The enteric coating can be applied using conventional processes known to those skilled in the art.

[0057] In addition to the above, the present invention also encompasses the possibility that any of the probiotic, probiotic pharmaceutical or nutritional compositions of the present invention (as reflected in any aspect or embodiment described throughout this specification) may be administered to a subject together with other ingredients or compounds, but these are not part of the probiotic, probiotic pharmaceutical or nutritional composition of the present invention. In this sense, when the composition of the present invention is formulated as a nutritional composition, the nutritional composition may be a food product or may be incorporated into a food or food product intended for both human and animal consumption. Thus, in certain embodiments, the nutritional composition is selected from a food product (which may be a food product for specific nutritional purposes or a food product for medicinal purposes) and a nutritional supplement.

[0058] In the present invention, the term "nutritional composition" refers to a food product that provides nutrients to a subject consuming it, thereby beneficially affecting one or more bodily functions, resulting in better health and well-being. In the present invention, said nutritional composition is intended to alleviate, reduce, treat and / or prevent acne.

[0059] The term "supplement," which is synonymous with any of the terms "dietary supplement," "nutritional supplement," "food supplement," "alimentary supplement," or "alimentary complement," refers to a product or preparation intended to supplement the normal diet, consisting of a source of concentrated nutrients or other substances that have a nutritional or physiological effect. In the present invention, the "substance" that has a nutritional or physiological effect on an individual when the nutritional supplement is ingested is the microorganism L. rhamnosus CECT 30031 and / or Spirulina paraca BEA_IDA_0074B, which are part of any of the compositions of the present invention. Food supplements can be in single or combined forms and can be sold in dosage forms, i.e., capsules, pills, tablets, and other similar forms, powder sachets, liquid ampoules and dropper bottles, and other similar forms of liquids and powders designed to be taken as a single dose.

[0060] A wide range of nutrients and other elements may be present in nutritional supplements, including, inter alia, vitamins, minerals, amino acids, essential fatty acids, fiber, enzymes, plants and plant extracts. Their role is to supplement the supply of nutrients in the diet, so they should not be used as a substitute for a balanced diet, and the intake amount should not exceed the daily amount explicitly recommended by a doctor or nutritionist. Probiotic compositions may also be part of so-called "special group foods", i.e., foods that meet specific nutritional needs.

[0061] Examples of foods that may contain the compositions or probiotic compositions of the present invention (microorganisms L. rhamnosus CECT 30031 and / or Spirulina paraca BEA_IDA_0074B (or strains derived therefrom)) include, but are not limited to, animal feed, dairy products, vegetable products, meat products, snacks, chocolate, beverages, baby foods, cereals, fried foods, industrial bakery products, and biscuits. Examples of dairy products include, but are not limited to, products derived from fermented milk (e.g., yogurt or cheese) or non-fermented milk (e.g., ice cream, butter, margarine, or whey). Vegetable products include, but are not limited to, cereals and snacks, whether fermented (e.g., soy yogurt, oat yogurt, etc.) or non-fermented. Beverages may be, but are not limited to, non-fermented milk. In certain embodiments, the food product or food product is selected from the group consisting of fruit or vegetable juice, ice cream, infant formula, milk, yogurt, cheese, cultured milk, milk powder, freeze-dried or air-dried products (suitable for reconstitution with a liquid vehicle), cereal, baked goods, milk-based products, meat products, and beverages.

[0062] As will be appreciated by one of skill in the art, either the compositions of the present invention or the probiotic compositions may be formulated to form part of a personal care product or cosmetic product that is administered to a subject, for example, topically. Thus, in certain embodiments, either the compositions of the present invention or the probiotic compositions may be a personal care product or cosmetic product.

[0063] Non-limiting examples of personal care products include bar soaps, liquid soaps (e.g., hand soaps), hand sanitizers (including rinse-off and leave-on alcohol-based and water-based hand sanitizers), cotton swabs and pads, shaving creams, talcum powders, toilet paper, pre-surgical skin disinfectants, wet wipes, cleansing wipes, disinfectant wipes, body soaps, acne treatment products, antifungal diaper rash creams, antifungal skin creams, shampoos, conditioners, cosmetic deodorants, antibacterial creams, body lotions, hand creams, topical creams, aftershave lotions, toners, oral hygiene products, and sunscreen lotions. The compositions of the present invention can also be applied to wound care products, such as, but not limited to, wound healing ointments, creams and lotions, wound dressings, burn creams, bandages, tapes, and steri-strips, as well as medical supplies such as medical gowns, caps, masks, and shoe covers, surgical drops, etc. In one particular embodiment, the personal care product is a topical cream or gel.

[0064] Additionally, any of the compositions, probiotic compositions, pharmaceutical or nutritional compositions of the present invention (as reflected in any aspect or embodiment described throughout this specification) may include other microorganisms in addition to L. rhamnosus CECT 30031 and / or Spirulina paraca BEA_IDA_0074B. Thus, in certain embodiments, any of the compositions, probiotic, pharmaceutical or nutritional compositions of the present invention (as reflected in any aspect or embodiment described throughout this specification) may further include one or more microorganisms, particularly selected from the following group: Lactobacillus spp., Streptococcus spp., Bifidobacterium spp., Saccharomyces spp., and combinations thereof.

[0065] In another specific embodiment, any of the compositions of the invention is administered to a subject with food.

[0066] In another specific embodiment, any of the compositions of the present invention is administered to a subject by oral administration.

[0067] As will be understood by those skilled in the art, the microorganisms L. rhamnosus CECT 30031 and / or Spirulina paraca BEA_IDA_0074B (or any microorganisms derived therefrom) must be present in the probiotic, pharmaceutical, or nutritional compositions of the present invention (as reflected in any aspect or embodiment described throughout this specification) in an effective amount, particularly a therapeutically effective amount, to be effective in alleviating, reducing, treating, and / or preventing acne, particularly acne vulgaris, and more particularly comedones. In the present specification, an "effective amount" or "therapeutically effective amount" refers to the amount of a component or compound of the probiotic, pharmaceutical, or nutritional composition of the present invention sufficient to produce the desired effect when administered to a subject. The components or compounds of the probiotic, pharmaceutical, or nutritional compositions of the present invention (as reflected in any aspect or embodiment described throughout this specification) refer to the microorganisms L. rhamnosus CECT 30031 and / or Spirulina paraca BEA_IDA_0074B. A therapeutically effective amount will vary depending on, among other factors, the age, weight, general health, sex and diet of the subject, as well as the mode and time of administration, excretion rate or drug combination.

[0068] In another specific embodiment, the total concentration of the microorganisms L. rhamnosus CECT 30031 and / or Spirulina paraca BEA_IDA_0074B (or any microorganism derived therefrom) in any of the compositions of the invention is 10 3 cfu~10 12 cfu, especially around 10 9 In another particular embodiment, the administered dose of the microorganism L. rhamnosus CECT 30031 and / or Spirulina paraca BEA_IDA_0074B (or any microorganism derived therefrom) in the composition is 10 6 cfu / day ~10 12 cfu / day, especially 10 9cfu / day, and in another more particular embodiment the dosing regimen is at least once daily, particularly twice daily, more particularly three times daily, once with each meal intake (breakfast, lunch and dinner).

[0069] In another specific embodiment of any of the probiotic pharmaceutical or nutritional compositions of the invention (as reflected in any aspect or embodiment described throughout this specification), the concentration of L. rhamnosus CECT 30031 is at least thirty percent (30%) relative to the total concentration of microorganisms present in the composition, particularly at least 31%, 32%, 33%, 34% or 35% relative to the total concentration of microorganisms present in any of the compositions of the invention. In another more particular embodiment, the concentration of L. rhamnosus CECT 30031 is at least 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, preferably at least 60%, 65%, 70%, 75% or 80%, most preferably at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96% of the total amount of microorganisms present in any of the compositions of the invention.

[0070] In another embodiment of the present invention, either the probiotic medicine or the nutritional composition comprises 10 3 Colony forming units (cfu) ~10 12 colony forming units (cfu), more preferably at least about 10 6 cfu, most preferably at least about 10 9 cfu of L. rhamnosus CECT 30031.

[0071] In a preferred embodiment of the present invention, the concentration of L. rhamnosus CECT 30031 is between 90% and 95% relative to the total amount of microorganisms present in the composition of the present invention. Preferably, this concentration of L. rhamnosus CECT 30031 is between 10% and 95% relative to the total amount of microorganisms present in the composition of the present invention. 3 Colony forming units (cfu) ~10 12colony forming units (cfu), more preferably at least about 10 6 cfu, most preferably about 10 9 yielding an amount of cfu L. rhamnosus CECT 30031.

[0072] In one embodiment of the invention, the amount of S. paraca BEA_IDA_0074B is at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% of the total dry weight of the microorganisms present in the composition of the invention.

[0073] In a further embodiment, the composition of the invention comprises 2 mg to 20 mg, preferably 3 mg to 15 mg, most preferably 5 mg to 12 mg (dry weight) of S. paraca BEA_IDA_0074B.

[0074] In a further embodiment, any of the compositions of the invention comprises at least 0.5 mg of phycocyanin, preferably derived from S. paraca BEA_IDA_0074B, included in the composition.

[0075] In the context of the present invention, it should be understood that approximately 10% of the total dry weight of S. paraca BEA_IDA_0074B is composed of phycocyanin produced by the strain itself. Accordingly, any of the compositions of the present invention preferably contains at least 5 mg (dry weight) of S. paraca BEA_IDA_0074B, resulting in an amount of phycocyanin in the composition of at least 0.5 mg (dry weight).

[0076] In a preferred embodiment of either the probiotic pharmaceutical or nutritional composition of the present invention, the composition is provided in daily dosage units, each daily dosage unit containing at least 0.5 mg (dry weight) of phycocyanin derived from S. paraca BEA_IDA_0074B and at least about 10 6 cfu, preferably about 10 9 cfu of L. rhamnosus CECT 30031.

[0077] In a further embodiment of the present invention, the probiotic pharmaceutical or nutritional composition of the present invention further comprises at least one bulking agent and / or at least one anti-caking agent and / or at least one coating agent and optionally at least one opacifying agent.

[0078] Preferably, the at least one bulking agent of the present invention is maltodextrin, and / or the at least one coating agent of the present invention is gelatin, and / or the at least one opacifying agent is titanium dioxide.

[0079] The at least one anti-caking agent of the present invention may be selected from the group consisting of magnesium stearate, calcium stearate, silica, silicates such as sodium silicate or calcium silicate, talc, flour, starch, or combinations thereof.

[0080] In a preferred embodiment, the bulking agent is maltodextrin, the anti-caking agent is magnesium stearate, the coating agent is gelatin, and the opacifying agent is titanium dioxide.

[0081] Specifically, any of the probiotic compositions, probiotic pharmaceutical compositions, or nutritional compositions of the present invention (reflected in any aspect or embodiment described throughout this specification) can be used in a method for treating or preventing acne, particularly acne vulgaris, in a subject or individual, more specifically a method for alleviating, alleviating, treating, and / or preventing comedones, the method comprising administering an effective amount of the composition to the subject or individual, particularly orally, thereby treating or preventing the acne or comedones. As already indicated above, it is noted that oral liquid formulations used in the compositions of the present invention can be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups, or elixirs, or can be presented as dry products, such as lyophilized (freeze-dried) or air-dried products, which are reconstituted with water or other suitable liquid vehicles before use. Such liquid formulations can contain conventional additives, such as suspending agents, emulsifiers, non-aqueous vehicles (which may include edible oils), or preservatives. In a further embodiment, the method of the present invention for treating acne in a subject may further comprise the step of administering to the subject, sequentially, consecutively, or simultaneously, an effective amount of an additional agent, such as isotretinoin, salicylic acid, witch hazel, or benzoyl peroxide, a topical or oral retinoid, spironolactone, an oral contraceptive, azelaic acid, glycolic acid, a topical or oral antibiotic, a sulfa antibiotic, an spf / sunscreen, a moisturizer, or in other embodiments, a combination thereof. One skilled in the art will readily recognize that the components of any therapeutic composition may be adjusted and optimized based on the symptoms exhibited by the subject, their severity, and the potential for synergistic or antagonistic interactions between the components of such treatment without departing from the scope of the present invention.

[0082] In a further embodiment, the methods and compositions of the present invention are used as an adjuvant or supplemental treatment for acne, typically administered orally in an effective amount of any of the compositions of the present invention as defined herein, either alone or in combination (simultaneously, sequentially, or consecutively) with another compound(s) such as isotretinoin, salicylic acid, witch hazel, or benzoyl peroxide, topical or oral retinoids, spironolactone, oral contraceptives, azelaic acid, glycolic acid, topical or oral antibiotics, sulfa antibiotics, SPF / sunscreen, or moisturizers, particularly, but not limited to, topical or oral antibiotics, sulfa antibiotics, or hormonal agents for the treatment of acne, such as oral contraceptives or antiandrogens. Administration can be in one embodiment, a continuous therapy in a single unit dosage form, or in another embodiment, a single-dose therapy, as appropriate. Other embodiments of administration are effective in treating acne conditions. In other embodiments, any of the compositions of the present invention are used when symptomatic relief is specifically needed, or, in one embodiment, urgently needed. Finally, any of the compositions of the present invention may, in another embodiment, further be used as a continuous or preventative treatment.

[0083] Throughout the specification and claims, the word "comprises" and variations thereof are not intended to exclude other technical features, additives, ingredients, or steps. Other objects, advantages, and features of the present invention will be apparent to those skilled in the art, partly from the description and partly from the practice of the present invention. The following examples and drawings are offered by way of illustration and are not intended to limit the scope of the present invention. [Example]

[0084] Example 1. Probiotic Blend Study Study products: probiotic blend and placebo The probiotic blend used in this first example had a concentration of at least 1 x 10 per gram. 9It is a maltodextrin-based formulation named Bths-03 containing a mixture of two strains containing cfu (colony forming units).

[0085] The following table contains details regarding the composition and function of the treatments used during this pilot study (Table 2):

[0086] [Table 2]

[0087] The composition was in the form of white gelatin capsules (specifically 30 capsules per product) of 570 mg each, each having dimensions of 21.7 mm±0.5%.

[0088] The placebo was prepared in the same manner as the experimental drug, containing 30 mg of tapioca maltodextrin per capsule.

[0089] Methods for producing the blended products studied The lyophilized probiotic L. rhamnosus and lyophilized Spirulina were mixed prior to encapsulation. The mixing was carried out in a clean room at low temperatures to maintain the viability of the probiotic strains. After mixing, the capsules were encapsulated.

[0090] Information about dosage and how to take the product 1 capsule daily for 12 weeks.

[0091] The capsules were of a suitable size for easy swallowing.

[0092] The capsules were easy to open and suitable for dispersing their contents in a glass of water.

[0093] Indications under study Improvement of acne symptoms in adolescents and adults.

[0094] Study design A 12-week randomized, double-blind, placebo-controlled study using selected probiotic-containing products in the active arm.

[0095] population Adolescent and adult men and women aged 12-30 years with moderate acne were randomly assigned in a 1:1 ratio to two study arms: experimental (probiotic) and placebo.

[0096] Research objectives Main purpose To determine whether supplementation with the probiotic mixture Bths-03 has a beneficial effect and reduces acne symptoms in adolescents and adults.

[0097] Secondary Objectives To assess patient adherence to and experience with treatment; To determine the safety of the study treatment.

[0098] Research plan and methods Study design A double-blind, placebo-controlled pilot study of the efficacy of the orally administered probiotic Bths-03 in patients aged 12 to 30 years with moderate acne over a 12-week study treatment period.

[0099] Patient selection criteria The study enrolled adolescent and adult men and women aged 12 to 30 years with moderate acne; healthy patients were not included. To participate in the study, subjects had to meet all of the inclusion criteria detailed below and not meet any of the exclusion criteria.

[0100] Inclusion criteria 1. Signing of informed consent by the patient (and their legal guardian in the case of a minor) in accordance with the Clinical Trial Law. 2. Ages 12 to 30 years old. 3. Moderate acne according to the AGSS scale (Acne Global Severity Scale) and / or GAGS (Global Acne Grading System).

[0101] Exclusion criteria 1. Contraindications to any of the ingredients of the product under study. 2. Use of topical or systemic antifungal and antibiotic drugs in the past 2 weeks. 3. Probiotic intake in the past 2 months. 4. Use of systemic retinoids in the past 6 months.

[0102] Study variables Main Variable GAGS index score (Global Acne Scoring System) at the start of treatment, 6 weeks, and 12 weeks.

[0103] Secondary Variables AGSS Scale (Acne Global Severity Scale) Number of acne lesions assessed by digital photography Patients' views and subjective assessments Studying patient adherence to treatment Monitoring for possible adverse effects of treatment

[0104] Structure and research results The total intervention period for each patient in the study was 12 weeks, during which they received either the study product Bths-03 or a placebo. The study consisted of three visits where patients were seen: an initial visit (week 0), an interim visit (week 6), and a final visit (week 12). The following diagram illustrates the details of each study visit (Table 3):

[0105] [Table 3]

[0106] Collection and interpretation of adverse effects Adverse events were collected at the second and third study visits according to the information provided by the patients.

[0107] Similarly, there was a procedure to be followed in case of serious adverse events (AAG), but this event did not occur during the course of the study. According to the study, AAG, regardless of dose, was considered to cause: death, Immediately threatening the subject's life, resulting in hospitalization or prolonged hospitalization; causing a persistent or significant impairment or disability (the inability to properly perform life functions); Causing birth defects or congenital malformations; In women, pregnancy is also considered an AAG, and Suspected adverse events that are considered to be important and clinically relevant even if they do not meet the above criteria.

[0108] statistical analysis Number of patients enrolled in the study A sample size of 25 patients was expected to participate in the study. When comparing the two treatment groups, the main factors considered for sample calculation were: 1. The magnitude of the difference in a measured variable between two groups that can be detected. 2. The amount of variability present in the factor of interest being analyzed. 3. The expected "p" value (alpha risk) to be used as a measure of statistical significance. 4. The safety of detecting a statistically significant difference, assuming it exists (β risk).

[0109] Randomization method The clinical study was double-blind and randomized, i.e., both the investigators and the patients were unaware of the product (placebo or treatment) each participating patient was taking.

[0110] To maintain this randomization and avoid bias in the evaluation of participating patients, each patient was assigned to a treatment using a randomization table, which assigned a random code to each product to prevent its identification. These tables were maintained by a responsible person outside the study, who was contacted by the researchers each time a new patient was included.

[0111] Evaluation and analysis of results Demographic data and other basic characteristics In this study, the subjects' data were collected in a data collection notebook (CRD), and the data were entered into a database created using the statistical program SPSS v22.

[0112] Clinical data management was performed according to ongoing data cleansing rules and procedures, such as elimination of errors and discrepancies of the same, to ensure data integrity. Adverse events and concomitant medication terms were coded according to the Medical Dictionary for Reproductive Health (MedDRA).

[0113] The Student's t-test was used to test for statistically significant differences between the means of variables associated with each treatment group. In cases where the normality condition was clearly violated, the non-parametric Mann-Whitney U test was used.

[0114] Safety analysis Adverse events were analyzed by their number in each intervention group, their severity, and their relationship to the study product.

[0115] Missing, absent, or atypical data A 10% loss to follow-up rate was estimated in the study design. All patients who completed the study (protocol) scheduled visits were included in the final data analysis (evaluable subjects), and all other patients were excluded from the analysis after withdrawal from the study without being replaced by new patients.

[0116] result Characteristics of the population being studied A total of 25 patients who met the inclusion criteria and no exclusion criteria were evaluated during the study. The table below (Table 4) shows the general characteristics of this entire population included in the study.

[0117] [Table 4]

[0118] A total of 32 patients were evaluated during the course of the study, of which 7 were ultimately not included because they did not meet the protocol-defined inclusion and exclusion criteria and / or refused to participate in the study. From the total number of patients, the remaining 25 participating patients were randomized at enrollment and assigned to one of the treatment groups (probiotic or placebo). During this process, 12 patients were placed in the probiotic group and 13 in the placebo group, resulting in an even distribution of patients. During the study, a total of 1 patient (placebo) voluntarily withdrew and / or was lost to contact during the intervention. The consort diagram used in this study is shown in Figure 1.

[0119] When analyzing the data for these variables at baseline, no significant differences in characteristics were found between the two groups, and therefore adequate randomization was achieved. These data are shown in Table 5 below:

[0120] [Table 5]

[0121] Efficacy analysis Main variable: GAGS (Global Acne Scoring System) index Data for the primary variables are shown in Figure 2. It is noted that the mean reduction in the GAGS index (primary outcome) was 61% in the probiotic group and 24% in the placebo group relative to the values ​​shown. The difference at the end of the study compared to the values ​​shown was 61.8% in the probiotic group and 35.4% in the placebo group.

[0122] Secondary Variables AGSS (Acne Global Severity Scale) Index Data for this secondary variable are shown in Figure 3. The difference at the end of the study for the variable "Percentage of patients with improvement in two or more categories" was 58% in the probiotic group and 33% in the placebo group. The within-group difference was statistically significant in the probiotic group but not in the placebo group.

[0123] Number of inflamed acne lesions This variable was assessed by high-resolution digital photography. An interim analysis of the study, established 6 weeks after intervention, showed a 47.58% improvement in this variable in the probiotic group compared with a 19.37% improvement in the placebo group. Data regarding this variable at the end of the study are shown in Figure 4. The difference at the end of the study was 76.5% in the probiotic group and 23.38% in the placebo group. The difference between the groups was clinically and statistically significant (p<0.05).

[0124] Patients' views and subjective assessments In this variable, an interim analysis of the study established six weeks after the intervention showed an improvement of 11.67% in the probiotic group compared with a 3.33% improvement in the placebo group.

[0125] At the end of the study, the difference was 29.58% in the probiotic group and 4.75% in the placebo group. Data for this variable at the end of the study are shown in Figure 5.

[0126] Safety analysis and side effects The number, severity and relationship of events to the product. The number of adverse events was very low and not all of them were attributable to the intervention product. The final preparation of the product was well tolerated and at the interim analysis of the study established 6 weeks after the intervention, treatment adherence was 82.5% in the probiotic group and 86.67% in the placebo group. Data for this variable at the end of the study were 86.25% in the probiotic group and 86.67% in the placebo group.

[0127] Example 2. Trifurcation study Strains L. rhamnosus CECT 30031 and S. paraca BEA_IDA_0074B were each evaluated separately for the treatment of acne vulgaris. To this end, a small pilot study was conducted with five patients in each of three arms of the study using L. rhamnosus CECT 30031, S. paraca BEA_IDA_0074B, or CECT 30031 and S. paraca BEA_IDA_0074B. Specifically, a total of 15 volunteers with acne vulgaris were selected and randomized in a 1:1:1 ratio to one of the following three interventional treatments: 1. L. rhamnosus CECT 30031, one capsule per day containing 100 mg of probiotic strain at a total dose of 10E9 cfu / administration. 2. S. paraca BEA_IDA_0074B, one capsule per day containing a total of 100 mg of probiotic strains at a total dose of 10E9 cfu / administration. 3. A combination of two probiotic strains at the same doses as used individually.

[0128] The intervention lasted 12 weeks, during which patients were evaluated at the start of treatment and at 1, 2, and finally 3 months after the end of the intervention. At each visit, the response to treatment was assessed using an activity index.

[0129] The results of the three-branch study are summarized below: These outcomes include the difference, for each variable, between the value obtained at the end of the intervention study and the value obtained at the beginning, before treatment began. The results for each of the three intervention groups are listed here:

[0130] Strain 1. L. rhamnosus CECT 30031 (when used individually) IL-10: Increased 23% from baseline values ​​during the study. Blood lipopolysaccharide: 14% decrease. DNA in blood: 2% decrease. GAGS: 24% improvement. AGSS: Improved in 1 of 5 cases. Inflammatory spots: Decreased in 23% of cases.

[0131] Strain 2. Spirulina paraca (when used individually) IL-10: Increased 11% from baseline values ​​during the study. Blood lipopolysaccharide: Decreased by 6%. DNA in blood: 3% decrease. GAGS: 13% improvement. AGSS: No improvement. Inflammatory lesions: Decreased in 11%.

[0132] Blend with L. rhamnosus CECT 30031 and Spirulina paraca (when used together) IL-10: Increased 36% from baseline values ​​during the study. Blood lipopolysaccharide: 42% decrease. DNA in blood: 68% decrease. GAGS: 56% improvement. AGSS: Improved in 3 out of 5 cases. Inflammatory lesions: Decreased in 57%.

[0133] In conclusion, this study illustrates how the combined use of both strains potentiates and therefore significantly improves the efficacy compared to the efficacy of each of the individual strains.

[0134] Example 3. Comparative analysis In this example, a probiotic composition containing S. paraca BEA_IDA_0074B strain was compared with a probiotic composition containing a known strain (strain 1. L. rhamnosus SP1) specifically disclosed in "Fabbrocini G, Bertona M, Picazo O, Pareja-Galeano H, Monfrecola G, Emanuele E. Supplementation with Lactobacillus rhamnosus SP1 normalizes skin expression of genes implicated in insulin signaling and improves adult acne. Benef Microbes. 2016; 7 (5): 625-630. doi: 10.3920 / BM2016.0089." Such known strains have been considered effective in the prior art for the treatment of acne vulgaris (see Table 1 above). The design, intervention products, and results of this study are summarized below.

[0135] Study variables GAGS index score (Global Acne Scoring System) at the start of treatment and at 12 weeks. AGSS scale (Acne Global Severity Scale) at the start of treatment and at 12 weeks. The number of inflamed acne lesions at the start of treatment and at 12 weeks was assessed by digital photography.

[0136] Structure and research results The total intervention period for each patient in the study was 12 weeks, during which they received one of the study products: one group of five patients received L. rhamnosus sp1, a second group of another five patients received S. paraca BEA_IDA_0074B, and finally, a third group of five patients received a mixture of L. rhamnosus sp1 and S. paraca BEA_IDA_0074B. The study consisted of two visits where patients were seen: the first visit (week 0) and the final visit (week 12).

[0137] result Main Variable GAGS (Global Acne Scoring System) Index It is noted that the mean reduction in GAGS index values ​​(difference at the end of the study compared to the value before the start of treatment) was 15% in the group given L. rhamnosus SP1, 12% in the patients in the S. paraca BEA_IDA_0074B group, and 12% in the group of 5 patients given the combined composition of L. rhamnosus SP1 and S. paraca BEA_IDA_0074B.

[0138] Secondary Variables AGSS (Acne Global Severity Scale) Index The number of patients who improved during treatment (comparing results at the start with results at the end of the study) was 1 in 5 (20%) in the L. rhamnosus SP1 group, 0 in 5 (0%) in the S. paraca BEA_IDA_0074B group, and finally 1 in 5 (20%) in the mixture of L. rhamnosus and S. paraca BEA_IDA_0074B group.

[0139] Number of inflamed acne lesions This variable was assessed by high-resolution digital photography. For this variable, a 17% reduction in the number of inflamed lesions was observed in the L. rhamnosus sp. group (comparing the number of lesions at the end of the study with the value at the beginning of the study). The reduction was 9% in the S. paraca BEA_IDA_0074B group and 12% in the group of patients given a mixture of L. rhamnosus and S. paraca BEA_IDA_0047B.

Claims

1. a. the Lactobacillus rhamnosus strain deposited under the Budapest Treaty at the Spanish Type Culture Collection (CECT) under accession number CECT 30031; b. Arthrospira platensis (S. paraca) strain deposited at Banco Espanol de Algas under accession number BEA_IDA_0074B; 10. A probiotic composition for use in the alleviation, reduction, treatment and / or prevention of acne in a subject in need thereof, comprising:

2. 10. The probiotic composition of claim 1, in the form of an air-dried or lyophilized (freeze-dried) powder.

3. 3. The probiotic composition according to claim 1 or 2, which is a formulation for administration in liquid or solid form.

4. 3. The probiotic composition according to claim 1 or 2, which is a formulation for oral administration.

5. The probiotic composition according to any one of claims 1 to 4, which is a pharmaceutical composition comprising a pharmaceutically acceptable excipient.

6. The probiotic composition according to any one of claims 1 to 4, which is a nutritional composition or a nutritional supplement.

7. 7. A nutritional composition or nutritional supplement according to claim 6 or a pharmaceutical composition according to claim 5, wherein the composition is in the form of a formulation selected from the group consisting of tablets, lozenges, confectioneries, chewable tablets, chewing gum, capsules, sachets, powders, granules, coated particles or coated tablets, tablets, and gastro-resistant tablets and capsules, and dispersible strips and films, or in the form of a liquid formulation selected from the group consisting of oral solutions, suspensions, emulsions and syrups.

8. 8. The probiotic composition of any one of claims 1 to 7 for use in treating acne in a subject in need thereof, wherein an effective amount of the composition is orally administered to the subject.

9. 9. The probiotic composition of any one of claims 1 to 8 for use in treating acne in a subject in need thereof, wherein at least one additional compound is administered to the subject in an amount effective to treat acne, the probiotic composition acting as an adjuvant.

10. 10. The probiotic composition according to any one of claims 1 to 9, wherein the acne is selected from the group consisting of acne vulgaris, acne rosacea, acne conglobata, acne fulminans, gram-negative folliculitis and facial pyoderma.

11. The probiotic composition of claim 10, wherein the acne is acne vulgaris.

12. 1. A probiotic composition comprising the Lactobacillus rhamnosus strain deposited under the Budapest Treaty at the Spanish Type culture Collection (CECT) under accession number CECT 30031 for use in the alleviation, reduction, treatment and / or prevention of acne in a subject in need thereof.

13. 13. The probiotic composition of claim 12, wherein the acne is selected from the group consisting of acne vulgaris, acne rosacea, acne conglobata, acne fulminans, gram-negative folliculitis, and facial pyoderma.

14. 14. The probiotic composition according to claim 12 or 13, adapted to be orally administered to a subject in need of said alleviation, relief, treatment and / or prevention.

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