Microbiota modulating cosmetic complex, cosmetic composition, uses for microbiota modulating cosmetic complex and method for cosmetic treatment

Abstract

The present invention relates to a microbiota modulating cosmetic complex comprising a combination of the active ingredients (a) recombinant silk protein, (b) xylitol phosphate ester, and (c) caffeine, as well as to cosmetic compositions containing said microbiota modulating cosmetic complex, uses therefor and a cosmetic method, the aim being to promote skin microbiota equilibrium and hair and scalp health.

Description

[0001] “MICROBIOTA MODULATING COSMETIC COMPLEX, COSMETIC COMPOSITION, USES OF THE MICROBIOTA MODULATING COSMETIC COMPLEX AND METHOD FOR COSMETIC TREATMENT” FIELD OF THE INVENTION

[0002] [1] The present invention relates to a cosmetic microbiota modulating complex comprising the combination of the active ingredients: (a) recombinant silk protein, (b) xylitol phosphate ester and (c) caffeine, as well as cosmetic compositions containing the cosmetic microbiota modulating complex, its uses and method, aiming to promote the balance of the skin microbiota, and the health of the hair and scalp.

[0003] EARLIER ART

[0004] [2] Microbiota is the set of microorganisms present in a given ecosystem, such as the skin, hair or scalp.

[0005] [3] The skin microbiota is composed primarily of bacteria, but also includes viruses, fungi, and protozoa. The microorganisms present in the skin microbiota are determined by environmental conditions such as anatomical location, humidity, sebum and sweat production, oxygen concentration, pH, among others, as well as biological factors such as hormonal fluctuations and the individual's age.

[0006] [4] The hair or scalp microbiota, in turn, is a community of microorganisms that develop on the scalp and may be present on the hair shaft due to its proximity to the scalp. It is composed of bacteria and fungi, which feed on sebum and keratinocytes (dead skin cells).

[0007] [5] The microbiota is responsible for several physiological functions, such as the production of nutrients or defense peptides, modulation of the immune response and, in the case of the hair microbiota, it also has the function of protecting and maintaining healthy hair.

[0008] [6] Microorganisms can attach to the skin by attaching to structures in the skin or hair, such as carbohydrates or cell receptors, within skin cells, or organized into colonies bound together by a biofilm.

[0009] [7] Biofilm forms a microenvironment with a different pH, oxygen concentration, nutrients, and metabolites than that of the skin. It also facilitates bacterial communication through quorum sensing and protects the microbiota from endogenous and exogenous antimicrobial agents and the individual's immune system.

[0010] [8] Human skin contains approximately 1,000 bacteria for every cell. Genetic sequencing studies show that bacteria exist not only on the surface of the skin, but also in sweat and sebaceous glands, follicles, skin invaginations, and even in deeper layers of the skin, such as the dermis.

[0011] [9] The most common genera in the superficial layers of the skin are Staphylococcus, Cutibacterium, Micrococcus and Corynebacteria.

[0012] These bacteria are distributed across different regions of the skin depending on the microenvironment. Staphylococcus and Corynebacteria, for example, are the most abundant genera on the skin and persist throughout life. Cutibacterium, which are anaerobic, are present in the sebaceous glands. Corynebacteria are present in the armpits and are linked to the production of bad odor. Fungi of the genus Malassezia are present on the scalp and are linked to seborrheic dermatitis and dandruff.

[0013]

[0011] Within the topic of scalp health, hair loss is a cosmetic issue with a multifactorial etiology. Hair loss can occur due to breakage, telogen effluvium or temporary hair loss, diffuse alopecia, and even androgenic alopecia.

[0014]

[0012] Hair loss due to breakage is non-pathological and occurs due to external damage to the hair. It usually results from continuous and / or acute physical and / or chemical aggressions that cause substantial mass loss in areas of the cortex and cuticle. The damage generated creates areas of greater fragility to breakage, and thus, with the least physical stress, hair breakage is observed in any region along the length of the hair fiber. This condition can be easily recognized by phenomenological observation, in which hair loss resulting from breakage is observed when hands or tools that facilitate detangling and combing pass through it. Diffuse alopecia, also known as diffuse baldness, refers to hair loss of unnatural and cyclical origin and affects both the hair and the scalp.And finally, androgenic alopecia, also called permanent baldness, also refers to unnatural hair loss, but it is not cyclical and affects both the hair and the scalp.

[0015]

[0013] The classic mechanisms associated with diffuse hair loss have hormonal and inflammatory bases. It is known that the increase in the STAT3 protein reduces the anagen phase and decreases hair growth, the increase in the DHT hormone miniaturizes the follicle, causing hair thinning, and the increase in the IL1 b gene worsens follicle anchoring, reducing blood flow and accelerating hair loss.

[0016]

[0014] Human skin, in general, offers the microbiota a dry, slightly acidic environment with only proteins and lipids available.

[0017]

[0015] The balance of the microbiota involves adhesion mechanisms, interaction with the individual's immune defense system, changes in the microenvironment, nutrient consumption and production of antimicrobial agents.

[0018]

[0016] The superficial layers of the skin are constantly shedding, removing part of the microbiota. The ability to adhere to viable skin cells ensures the persistence of microorganisms. Bacteria bind to membrane receptors on cells, such as those of the toll-Hke family that recognize patterns associated with bacteria (TLR 1-10), mannose receptors, or NOD- / / e receptors. These receptors trigger immune pathways that discriminate between pathogenic and beneficial bacteria, stimulating appropriate responses. The composition of the microbiota teaches the immune system to more accurately discriminate between beneficial bacteria, modulating inflammatory responses and reducing the skin's sensitivity to external factors such as variations in temperature, humidity, and UV radiation exposure.

[0019]

[0017] In addition, the microbiota produces lactic acid which, together with free fatty acids (from sebum), acidic amino acids (from sweat) and urocanic and pyrrolidone carboxylic acids (from the keratinization process of corneocytes) maintain a more acidic skin pH, restricting microorganisms present in the microbiota. Lipids are the main source of carbon used by the skin microbiota.

[0020]

[0018] Variations in age, sex and hormonal cycle interfere with the availability of nutrients for the microbiota, which can lead to fluctuations in the composition of microorganisms.

[0021]

[0019] The skin's role as a selective barrier against the external environment is greatly facilitated by antimicrobial peptides (AMPs), produced by both keratinocytes and the skin microbiota. More than 2,000 molecules with antimicrobial activity have been described, produced by microorganisms and cataloged in public and private databases. These include α-defensins (hBDs), cathelicidins, C-type lectins, dermicidins, and others. Most AMPs are composed of small peptides of approximately 2–15 kDa. An imbalance (excess or lack) of AMPs in the skin is associated with conditions such as atopic dermatitis, rosacea, and psoriasis. Some skin-produced factors, such as vitamin D and PTH, regulate AMP synthesis. AMPs are important for protecting the skin against pathogens, controlling the microbiota population and limiting contact between keratinocytes and the microbiota.The activity of AMPs is generally broad-spectrum, affecting both Gram-positive and Gram-negative bacteria and, in some cases, fungi, protozoa and viruses.

[0022]

[0020] The microbiota also produces other substances, such as hyaluronic acid, produced by Streptococcus and some Lactobacillus, which is a constituent of the extracellular matrix that fills the dermis, giving shape to the skin and stimulating the production of [3-defensin 2 (hBD2).

[0023]

[0021] Sphingomyelinase is produced by keratinocytes, Streptococcus and Lactobacillus, and is an enzyme that participates in the synthesis of ceramides that are essential for maintaining the skin barrier.

[0024]

[0022] Lipoteichoic acid (LTA) is the structural component of Gram-positive bacteria such as Staphylococcus, Lactobacillus and Bifidobacteria, which stimulates the production of antimicrobial peptides such as β-defensins (hBD) and cathelicidins.

[0025]

[0023] Peptidoglycans are structural components of the cell wall of Gram-positive bacteria, which modulate the individual's immune response, stimulating the production of IL-8 by keratinocytes and recruiting phagocytes to infected sites.

[0026]

[0024] Lactic acid, alpha hydroxy acid, produced by Lactobacillus and Bifidobacteria, stimulates the skin peeling process, weakening cell adhesions, stimulating the production of ceramides by keratinocytes, strengthening the skin barrier and composing the NMF, which retains skin moisture.

[0027]

[0025] Acetic acid, produced by Lactobacillus and Bifidobacteria, has antibacterial activity against S. aureus and P. aeruginosa.

[0028]

[0026] The main representatives of the skin microbiota include Staphylococcus epidermidis, a major component of the skin, which is halotolerant and capable of growing in environments with low water activity. It is generally harmless to the skin, but can cause nosocomial infections through colonization of foreign bodies such as catheters and implants in predisposed and immunocompromised individuals. These bacteria are extremely resistant to AMPs, do not attack keratinocytes, and produce AMPs (epidermin, Pep5, epilancin K7, and epicidin 280) that control the populations of S. aureus and Streptococcus.

[0029]

[0027] However, removal of S. epidermidis from the skin, for example, through excessive use of topical antibiotics, leaves the individual immunocompromised and highly susceptible to infections by pathogens.

[0028] In addition, they synthesize proteases, keratinases, lipases, and nucleases that participate in the physiological turnover of the skin. They synthesize soluble phenolic modulins (PSMs), proteins with surfactant and antibacterial properties selective for S. aureus and Streptococcus, by dissolving the biofilm formed by these pathogenic bacteria.

[0030]

[0029] Staphylococcus aureus is found on healthy skin. It is the main pathogen of human skin, responsible for folliculitis, boils, subcutaneous abscesses, meningitis, endocarditis, and septicemia. Conditions such as atopic dermatitis, viruses (Herpes simplex type I or Papillomavirus), and opportunistic fungi (Trichophyton rubrum) predispose the skin to S. aureus infections.

[0031]

[0030] The increasing number of strains resistant to antibiotics such as metacycline and vancomycin makes S. aureus a serious problem in hospital settings. It synthesizes bacteriocins such as staphylococcin 462, which inhibit other strains of S. aureus.

[0032]

[0031] Corynebacteria jeikeium and Corynebacteria striatum are found on healthy skin, being the most common microorganisms after S. epidermidis. They are found in the axillary, inguinal, and perineal regions due to their high tolerance to salt concentrations. They adhere to keratinocytes at an estimated rate of 25 bacteria per cell through fibronectin receptors. They produce bacteriocins such as lacticidin Q, which is responsible for the synthesis of dermicidin and cathelicidin present in sweat, providing skin defense.

[0033]

[0032] C. acnes, present in the healthy microbiota in areas such as the face and back, is a saprophytic, anaerobic microorganism found primarily in the sebaceous glands and associated with manifestations such as folliculitis and acne vulgaris. Its main energy source is the lipids and fatty acids present in sebum. The presence of porphyrin makes C. acnes highly sensitive to UV radiation and stimulates the production of IL-8 and IL-12 by keratinocytes, creating a pro-inflammatory microenvironment.

[0033] Studies indicate that C. acnes is involved in the hypercorneification of the pilosebaceous duct, exacerbating inflammatory processes. Furthermore, it synthesizes acetic acid and propionic acid, as well as lipases and proteases. It also synthesizes several bacteriocins such as propionicin PLG-1, jensenin G, propionicin SM1, SM2, T1 and acnein, with activity against several lactic acid bacteria, Gram-negative bacteria, fungi and molds.

[0034]

[0034] Malassezia furfur is a fastidious fungus present in healthy microbiota in areas such as the scalp and back, primarily responsible for opportunistic infections such as seborrheic dermatitis, dandruff, and tinea versicolor (or pityriasis). Its main source of carbon is free fatty acids. Furthermore, it synthesizes an enzyme capable of degrading the cell walls of microorganisms, contributing to skin protection against pathogens.

[0035]

[0035] Microbiota imbalance can lead to seborrheic dermatitis, acne vulgaris, atopic dermatitis, among other diseases.

[0036]

[0036] The skin's microbiota is constantly removed by the natural process of desquamation and skin renewal. Therefore, the ability of MOs to adhere to viable skin cells, including in the deeper layers such as the dermis, is a competitive advantage.

[0037]

[0037] The presence of microbiota modulates the skin's inflammatory response, reducing sensitivity to factors such as temperature variations, humidity and exposure to UV radiation.

[0038]

[0038] The microbiota produces lactic acid, which helps maintain a more acidic skin pH, antimicrobial peptides (AMPs) that protect the skin from pathogens and whose imbalance is related to pathologies such as dermatitis, rosacea and psoriasis, help regulate the production of vitamin D and are involved in the production of extracellular matrix proteins such as hyaluronic acid and enzymes important for maintaining the skin barrier such as sphingomyelinase.

[0039]

[0039] Staphyloccocus epidermidis is the majority microorganism in the skin microbiota and remains so even during aging. S. epidermidis is harmless to keratinocytes and produces AMPs (epidermin, Pep5, epilancin K7, soluble phenolic modulins -PSMs- and epicidin 280) that control the populations of S. aureus and Streptococcus, protecting the skin.

[0040]

[0040] S. epidermidis synthesizes proteases, keratinases, lipases and nucleases that contribute to the physiological turnover of the skin and play a role in maintaining skin health, protecting it from harmful opportunistic microorganisms and favoring physiological mechanisms of skin renewal.

[0041]

[0041] In the scalp region, there are hair follicles with openings that contribute to a significant increase in the surface area susceptible to colonization by microbiota. The follicle is filled with sebum, debris, and microorganisms. Regarding the bacterial population present on the scalp, the population of bacteria of the genus Cutibacterium spp. (mainly Cutibacterium acnes) and the genus Staphylococcus spp. are the most abundant. The population of the genus Burkholderia spp. and Lawsonella can also be found. In healthy volunteers, the bacterial population is predominantly composed of bacteria of the phyla Actinobacteria, Firmicutes, and Proteobacteria, with organisms of the genus Cutibacterium spp. (mainly Cutibacterium acnes) and the genus Staphylococcus spp.

[0042]

[0042] Analysis of the microbiome of patients with androgenetic alopecia, the most common form of hair loss in men, demonstrated a predominance of Burkholderia spp in the medial compartment of hair follicles, both in patients and healthy volunteers. In patients with alopecia, dysbiosis of miniaturized hair follicles was observed, with enrichment of Cutibacterium acnes.

[0043]

[0043] Populations of the genera Lawsonella, Acinetobacter, and Prevotella can be found.

[0044]

[0044] In relation to the fungal population, the genus Malassezia spp. stands out as the most abundant.

[0045]

[0045] Disruption of the scalp microbiota by fungi and bacteria can lead to dysbiosis in the region, such as favoring local inflammation of the scalp skin.

[0046]

[0046] In relation to the fungal population, the genus Malassezia spp. stands out as the most abundant.

[0047]

[0047] Disruption of the scalp microbiota by fungi and bacteria can lead to dysbiosis in the region, such as favoring local inflammation of the scalp skin.

[0048]

[0048] In the case of hair, the microbiota is often deregulated by changes in hair pH (from progressive brushing, straightening, coloring, etc.), antibiotic treatments, pollution, as well as the use of products containing ingredients such as sulfates.

[0049]

[0049] Some evidence has begun to suggest an important role for the microbiota in hair loss, and as a result, many products have been developed that associate microbiota control with hair loss. Among these cosmetic products, we can cite as an example an anti-hair loss and anti-dandruff shampoo formulation for men that combines zinc pyrithione and Defenscalp to rebalance the scalp microbiome. However, zinc pyrithione indiscriminately eliminates the scalp microbiota, causing a rebound effect that causes dandruff to return more severely after use, in addition to leaving hair strands dry. The addition of Defenscalp is intended to alleviate these adverse effects, but it does not fully achieve its objective.

[0050]

[0050] There are numerous active ingredients available with suggested direct or indirect action on the microbiota, through topical or oral administration, such as acetic acid, diacetyl, lactic acid, hyaluronic acid, among others. And, although the components of the microbiota and their direct and indirect effects on the skin are known, the common use of antibacterials, which affect both beneficial and harmful bacteria present on the skin, persists. Their indiscriminate use weakens the skin's natural defenses and leaves it more vulnerable to harmful bacteria.

[0051] For this reason, new prebiotic and probiotic ingredients are still being researched.

[0051]

[0052] Prebiotics are ingredients that promote beneficial bacteria. Probiotics are microorganisms that interact with the skin's natural microbiota and stimulate its own defenses. Probiotics can also act as selective antibacterials, producing substances that control the proliferation of harmful bacteria.

[0052]

[0053] Numerous prior art publications indicate that prebiotic agents can selectively promote beneficial bacteria and, consequently, are candidates for use in cosmetic compositions for this purpose. However, despite screening and suggesting potential prebiotic ingredients, the prior art is limited in providing effective prebiotic ingredients with proven in vitro efficacy and an elucidated mechanism of action, particularly for topical administration. Therefore, research is still needed.

[0053]

[0054] Thus, there remains a need for active ingredients that have a proven microbiota-modulating effect on the skin, hair and scalp.

[0054] BRIEF DESCRIPTION OF THE FIGURES

[0055]

[0055] Figure 1 illustrates the results of combating hair loss and improving hair density and thickness after using the anti-hair loss lotion of the present invention.

[0056]

[0056] Figures 2A and 2B show the results obtained in relation to the diversity and abundance of fungi on the scalp after using the anti-hair loss lotion of the invention.

[0057]

[0057] Figures 3A and 3B show the results obtained in relation to the bacterial profile, diversity and abundance of bacteria on the scalp after using the anti-hair loss lotion of the invention.

[0058]

[0058] Figures 3C, 3D and 3E show the ratios between the genera of bacteria on the scalp after treatment with an anti-hair loss lotion of the invention.

[0059]

[0059] Figures 4A, 4B and 5 show the impact on the lipid composition of the scalp after using the anti-hair loss lotion of the invention. Figure 4A demonstrates that the lipid profile changes with treatment. Figure 4B demonstrates the variation in lipid concentration. And Figure 5 illustrates the correlation between microorganisms, lipids and clinical data (Spearman correlation with FDR adjustment).

[0060] DESCRIPTION OF THE INVENTION

[0061]

[0060] In a first aspect, the present invention relates to a cosmetic microbiota modulating complex comprising the combination of the active ingredients: (a) recombinant silk protein, (b) xylitol phosphate ester and (c) caffeine.

[0062]

[0061] The microbiota-modulating cosmetic complex of the invention surprisingly promotes the balance of the microbiota of the skin, hair and scalp, as well as the health of the hair and scalp.

[0063]

[0062] The present invention reveals a new approach associating the microbiota and scalp lipid profiles in hair loss.

[0064]

[0063] Through the characterization of the microbiota and the lipid profile of the scalp with alopecia, it was possible to evaluate the impact of the composition of the present invention, particularly anti-hair loss lotion on the microbiota (fungi and bacteria), and its comparison with healthy scalp correlating with clinical data.

[0065]

[0064] Thus, it was possible to verify the relationship between angiogenesis and inflammation markers that influence the balance of the microbiota and the lipid profile of the scalp, and the effect of anti-hair loss treatment on scalp health.

[0066]

[0065] A healthy scalp has certain characteristics such as hair that does not fall out, a natural hair growth cycle (on average 1 cm every 28 days), few inflammatory cells, microcirculation of the scalp feeding the follicle and a balance of microbiota and fungal species.

[0067]

[0066] On the other hand, a diseased scalp presents temporary hair loss, thinning of the strands and increased hair loss, inflammatory process, impaired microcirculation, imbalance of the microbiota and fungal species and increased production of FFAs (free fatty acids) and oiliness.

[0068]

[0067] The cosmetic complex comprising the combination of active ingredients: (a) recombinant silk protein, (b) xylitol phosphate ester and (c) caffeine, has the activity of modulating the microbiota, promoting the balance of the skin, hair and scalp, nourishing the beneficial microorganisms residing in the microbiota, strengthening the hair, reducing sensitivity and irritation with proven efficacy in vitro.

[0069]

[0068] In particular, the microbiota modulating cosmetic complex of the present invention acts as follows:

[0070] - Prolongs hair growth by keeping the anagen phase active;

[0071] - Inhibits hair thinning by reducing DHT while maintaining the hair's natural thickness; and

[0072] - Reduces inflammation by improving anchoring, microcirculation and nutrient flow.

[0073] - Balances the microbiota by controlling populations of Burkholderia spp.

[0074]

[0069] In particular, the recombinant silk protein is used in a concentration of 0.01-1%, in particular approximately 0.33%, relative to the total weight of the composition.

[0075]

[0070] According to one embodiment, the recombinant silk protein may be the recombinant ADF-4 protein which is known by the name Silkgel®.

[0076]

[0071] In particular, the xylitol phosphate ester is used in a concentration of 1-5%, in particular around 3%, relative to the total weight of the composition.

[0077]

[0072] According to the present invention, the xylitol phosphate ester used may be the ingredient Hygeaphos® obtained from the supplier Chemyunion.

[0078]

[0073] In particular, caffeine is used in a concentration of 0.01-1%, particularly around 0.65%, relative to the total weight of the composition.

[0079]

[0074] According to the present invention, caffeine can be obtained from the supplier Brenntag.

[0080]

[0075] The cosmetic compositions according to the present invention may be in the form of emulsions, solutions, gels, powders, pastes, among others, also comprising cosmetically appropriate vehicles for the chosen cosmetic form.

[0081]

[0076] The cosmetic compositions according to the present invention are intended for topical application.

[0082]

[0077] Surprisingly, it was found that the microbiota modulating effect provided by the cosmetic complex according to the present invention is not only efficient in reducing harmful bacteria and fungi on the skin, hair and scalp, favoring the beneficial bacteria in these regions, balancing their microbiota, but also provides balance and nutrition, providing advantageous cosmetic effects, combating hair breakage and loss and stimulating growth, increasing the density and thickness of the strands, without harming the microbiota of the skin and scalp.

[0083]

[0078] The microbiota-modulating topical cosmetic compositions according to the present invention may be made available in any cosmetic forms known to the person skilled in the art, suitable for application to the face, body, hair or scalp, for treatment or cleaning, including, without limitation, soaps, shampoos, elixirs, conditioners, sticks, tonics and lotions, among other forms.

[0084]

[0079] Cosmetically acceptable excipients may be selected from compounds known in the prior art. Without limitation, the excipients may be selected from the group comprising emollients, antioxidants, humectants, emulsifiers, surfactants, sensory or viscosity modifiers, preservatives, chelators, stabilizers, lubricants, thickeners, dispersants, solubilizers, and combinations thereof, among other cosmetically acceptable vehicles.

[0085]

[0080] The present invention also contemplates the use of a cosmetic microbiota modulating complex according to the present invention to reduce the diversity and abundance of fungi present on the scalp.

[0086]

[0081] In another aspect, the present invention contemplates the use of a cosmetic microbiota modulating complex according to the present invention, for maintaining the diversity and modulating the abundance of bacteria present on the scalp.

[0087]

[0082] An additional aspect of the present invention is a cosmetic treatment method that consists of applying to the skin, hair and / or scalp the microbiota modulating cosmetic complex or cosmetic composition defined in the present invention.

[0088]

[0083] The following examples, without imposing any limitation, illustrate the present invention. The examples aimed to: (1) evaluate the impact of the cosmetic product on the scalp microbiota profile with a diffuse alopecia process; (2) correlate clinical changes in the scalp with changes in the microbiota profile; and (3) correlate changes in lipid composition with changes in the microbiota profile.

[0089] EXAMPLES

[0090]

[0084] An anti-hair loss lotion composition (anti-hair loss night treatment elixir) was prepared:

[0091] Table 1 - Anti-Hair Loss Lotion

[0092] EXAMPLE 1 - Anti-hair loss treatment evaluation

[0093]

[0085] To assess the cosmetic effect attributed to the research objective of this descriptive study, a methodology was applied to evaluate the effectiveness of the products in stimulating new hair growth after 30, 60, 90, and 150 days of treatment with the experimental product. Forty research subjects who used the experimental product for up to 150 days were considered. The average age of the subjects was 37 ± 12 years. After recruitment, the subjects were instructed to discontinue use of any cosmetic product on their hair and scalp 48 hours before the start of the study. On the day of the study, the recruited subjects who came to the laboratory received clarification from the researcher regarding the study procedures, ethical and legal aspects, risks and benefits, medical support, and forms of reimbursement for participation costs. The subjects were also asked to sign two copies of the Informed Consent Form.Hair growth kinetics and density were assessed by obtaining scalp microimages using a microcamera (i-Scope USB, Moritex, JP) with a 30x objective and polarized light at baseline and after 30, 60, 90, and 150 days of treatment. Laser Doppler ultrasound was used to assess improvements in blood microcirculation at baseline and after 30, 60, 90, and 150 days of home use of the experimental product.

[0094]

[0086] The method used to evaluate the scalp microbiota was metagenomics, which consists of analyzing fragments of the genome of microorganisms, where a preserved region is evaluated, in bacteria called 16S rRNA and in fungi ITS as a universal marker. It is used in phylogenetic reconstruction to define its taxa, thus allowing the identification of each population in a given community with the aid of the lonChef System and Ion S5 following the NGS (Next-Generation Sequencing) protocols. The samples came from a swab rubbed on the scalp of individuals who used the product and were analyzed at baseline and after 30, 60, 90, and 150 days of treatment.

[0095] RESULTS

[0096]

[0087] The application of the composition of the present invention provides a double-action treatment, combating breakage and stimulating hair growth, as shown in the results illustrated in Figure 1 and summarized in the Table below:

[0097] Table 2 - Clinical efficacy results versus baseline time

[0098]

[0088] Therefore, based on the results, it can be seen that treatment with the anti-hair loss lotion (Table 1) of the present invention combats hair loss, improves hair density and contributes to improving the nutritional supply of the follicular units of the scalp.

[0099] EXAMPLE 2 - Assessment of the diversity and abundance of fungi on the scalp.

[0100]

[0089] The relative abundance of fungi is characterized by the predominance of the phylum Basidiomycota, which contains the genus Malassezia, and a minority by the phylum Ascomycota. This abundance profile is maintained with treatment, with small variations in the composition of minority fungi. The fungal diversity profile, calculated by the Shannon index, at 150 days of treatment remains close to the initial one, with significant changes occurring only between 30 and 90 days of treatment, observed by ANOVA + Tukey with Bonferroni adjustment (See Figures 2A and 2B).

[0090] Malassezia remains the main genus present at all times.

[0101] EXAMPLE 3 - Assessment of the diversity and abundance of bacteria on the scalp

[0102]

[0091] As can be seen in Figures 3A, the bacterial profile, in other words, the relative abundance of bacteria on the scalp, changes with treatment, with the phylum Actinobacteria increasing its predominance, with the genera Cutibacterium and Lawsonella being the main representatives observed in the samples. After 90 days of treatment, the relative abundance of the phylum Proteobacteria reduces, and bacteria from the Burkholderia-Caballeronia-Paraburkholderia group are the main representatives of this phylum in the samples (See Figure 3A).

[0103]

[0092] Additionally, when assessing the alpha diversity of genera using the Shannon index, a differentiation is observed after 90 days of treatment, with a significant reduction in diversity, as observed by ANOVA + Tukey with Bonferroni adjustment (See Figure 3B). In other words, the diversity of scalp bacteria decreases significantly with treatment time. Over time, the proportion of Burkholderia decreases, and other genera such as Cutibacterium, Staphylococcus, and Lawsonella begin to occupy a larger proportion. At 150 days, the proportions of Burkholderia increase slightly again.

[0104]

[0093] Furthermore, when evaluating the ratio between the major genera, significant differences were observed for the Cutibacterium-Burkholderia and Staphylococcus-Burkholderia ratios with the treatment, and the Cutibacterium-Staphylococcus ratio did not change with the treatment (ANOVA + Tukey with Bonferroni). See Figures 3C, 3D and 3E.

[0105] EXAMPLE 4 - Assessment of impact on scalp lipid composition

[0106]

[0094] A trial was conducted to evaluate the impact of cosmetic use on the lipid composition of scalps with diffuse alopecia. The experimental procedure for collecting sebaceous lipids from the scalp was performed using a non-invasive technique widely used in dermatology studies. It consisted of applying lipophilic strips to the volunteer's scalp and keeping them in contact with the skin for a standardized period of time and at controlled pressure. These strips (Sebutape™) have a special chemical characteristic that allows for the selective capture of sebaceous lipids without interfering with the biochemical composition of the collected samples. The standardized time and pressure minimize variability between collected samples, ensuring more reliable results. The strips were applied by trained professionals to minimize inter-observer variation and ensure procedure uniformity.After collection, the strips were stored under appropriate conditions to preserve the samples and were subsequently analyzed to determine the composition of sebaceous lipids. The method is considered safe and minimally invasive, with low risks and complications associated with sebaceous lipid sample collection.

[0107]

[0095] From the point of view of quantitative chemical analysis, a method using liquid-liquid extraction, derivatization, and GC-MS was developed to quantify different lipid classes in scalp samples. Initially, the lipid compounds were extracted from the adhesive tape using ethyl acetate as a solvent. After this step, these compounds were subjected to derivatization with N,O-Bis(trimethylsilyl)trifluoroacetamide (BSTFA) before injection into the GC-MS system. Calibration curves were constructed by diluting the standards within a linear range that meets their concentrations in the sample. Furthermore, a relative quantification approach was employed for other compounds present in the real samples, which were directly correlated with the standards of the monitored lipid classes.

[0108]

[0096] Additionally, a correlation was made between data on scalp sebum composition and the bacterial / fungal microbiota and hair loss.

[0109] RESULTS

[0110]

[0097] Treatment with the anti-hair loss lotion of the invention globally increases the lipids present in the scalp (see Figures 4A and 4B), demonstrating that the lipid profile changes with the treatment.

[0111]

[0098] The lipid profile correlates with the major genera of bacteria found and varies with the use of the anti-hair loss lotion (Figure 5). The hypothesis is that treatment with the anti-hair loss lotion modulates the lipid profile and disrupts the metabolism of enzymes necessary to maintain the population of Burkholderia spp.

[0112]

[0099] An inflammatory process, miniaturization of the follicle, excess consumption of lipids present in the scalp, an increase in the population of Burkholderia spp and a reduction in the population of Staphylococcus spp are observed in TO.

[0113]

[0100] At T30, a reduction in hair loss, an increase in lipid production and a reduction in the population of Burkholderia spp. are observed.

[0114]

[0101] The person skilled in the art will readily know how to evaluate, through the teachings contained in the text and the examples presented, the advantages of the invention and propose variations and equivalent alternatives for implementation, without departing from the scope of the invention, as defined in the attached claims.

Claims

CLAIMS 1. MICROBIOTA MODULATING COSMETIC COMPLEX, characterized by comprising the combination of the active ingredients (a) recombinant silk protein, (b) xylitol phosphate ester and (c) caffeine.

2. COSMETIC COMPLEX, according to claim 1, characterized by comprising: (a) from 0.01 to 1% of recombinant silk protein; (b) from 1 to 5% of xylitol phosphate ester; and (c) from 0.01 to 1% of caffeine, all percentages in relation to the total weight of the cosmetic composition.

3. COSMETIC COMPOSITION, characterized by comprising the microbiota-modulating cosmetic complex defined in either of claims 1 or 2 together with cosmetically acceptable excipients.

4. COSMETIC COMPOSITION, according to claim 3, characterized by being a topical composition.

5. COSMETIC COMPOSITION, according to claim 3 or 4, characterized by being in the form of an emulsion, solution, gel, powder and / or paste.

6. COSMETIC COMPOSITION, according to any one of claims 3 to 5, characterized by being a soap, shampoo, elixir, conditioner, stick, tonic and / or lotion.

7. COSMETIC COMPOSITION, according to claim 3, characterized in that the cosmetically acceptable excipients are selected from the group comprising emollients, antioxidants, humectants, emulsifiers, surfactants, sensory or viscosity modifiers, preservatives, chelators, stabilizers, lubricants, thickeners, dispersants, solubilizers, and combinations thereof.

8. USE OF THE MICROBIOTA MODULATING COSMETIC COMPLEX, as defined in either of claims 1 or 2, characterized by the reduction of the diversity and abundance of fungi present on the scalp.

9. USE OF THE MICROBIOTA MODULATING COSMETIC COMPLEX, as defined in either of claims 1 or 2, characterized by maintaining the diversity and modulating the abundance of bacteria present on the scalp.

10. METHOD FOR COSMETIC TREATMENT characterized by consisting of applying to the skin, hair and / or scalp, a cosmetic microbiota modulating complex, as defined in any one of claims 1 or 2, or a cosmetic composition, as defined in any one of claims 3 to 7.

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