Ribbed kapok flower extract enriched in polysaccharides

Abstract

The present invention relates to an extract from flowers of Ceiba trichlosta, preferably from the calyx of Ceiba trichlosta, a process for preparing it, and the extract obtained by said process. The present invention also relates to a composition comprising said extract, said composition being advantageously a cosmetic, pharmaceutical or dermatological composition. The present invention also relates to said composition or said extract for use in the prevention or treatment of diseases or pathological conditions of the skin, mucosae or hair and nails, and of imbalances or diseases associated with a microbial imbalance of the skin, mucosae, hair, nails and related cutaneous appendages. Finally, the present invention relates to a cosmetic care method for the skin, hair and nails or mucosae to improve their condition or appearance, said method comprising the application of said composition or said extract.

Description

Technical Field

[0001] This invention relates to an extract of the flower of *Bombax costatum*, which is rich in polysaccharides. The invention also relates to a cosmetic, pharmaceutical, or dermatological composition comprising this extract. Furthermore, the invention relates to a method for extracting the polysaccharide-rich *Bombax costatum* flower, and the extract obtainable by said method. The invention further relates to the use of this composition or extract in the prevention or treatment of diseases or pathological conditions of the skin, mucous membranes, or skin appendages, and in the prevention or treatment of vascular diseases. Finally, the invention relates to a method of care for the skin, skin appendages, or mucous membranes to improve their condition or appearance, said method comprising applying the composition or extract. Background Technology

[0002] Bone-fruited kapok

[0003] Bombax laurentii belongs to the family Bombacaceae (APG: Malvaceae). It is also known as Bombax andrieui or Bombax houardii. More commonly, it is called red-flowered kapok, cheese tree, red cotton tree, false kapok, forest kapok, Voaka (Moray), or Boumbou (Jura).

[0004] The genus *Bombax* is a pantropical genus containing eight species: two in Africa, five in Asia, and one in Oceania, extending as far as the Solomon Islands. In the past, the genus *Bombax* was defined much more broadly. *Bombax buonopozense* is sometimes considered the same species as *Bombax buonopozense*.

[0005] Uses of Bone-fruited Kapok

[0006] In Africa, the ribbed kapok is a fairly common edible tree: its leaves are dried and eaten like baobab leaves; its flowers, especially the calyx, are also eaten.

[0007] In Burkina Faso, the bark is used to stain teeth. The flowers are usually harvested for their fleshy calyxes, which are cooked and eaten as a vegetable. The leaves are also eaten as vegetables. The immature fruits, and sometimes the flowers, are added as a thickener to sauces. Young, immature fruits are also used to make beverages. The seed oil is edible. The flowers are highly valued as nectar.

[0008] Many parts of the tree have been used in traditional medicine to treat various ailments. Powdered root extracts are consumed in sauces or used as a bath to treat epilepsy. Bark preparations are applied to wounds to promote healing.

[0009] In Senegal and Sierra Leone, the diuretic properties are attributed to the stem and root bark.

[0010] The bark is also used to prepare medicines for treating trichomoniasis, amoebiasis, and other forms of dysentery. Bathing in stem bark extract is used to treat mental disorders. Powdered stem bark is used in pharmaceutical compositions as a fumigant for treating headaches. For treating headaches or toothaches, bark poultices can be placed on the head.

[0011] The leaves, along with other medicinal plants, are used to treat leukorrhea and diarrhea. An extract of crushed leaves is used as a beverage to address problems during childbirth. Repeated baths with an extract of crushed leaves are used to treat convulsions. A tea made from dried leaves is taken or applied topically to treat measles. In cases of severe swelling, a decoction of the leaves, stems, or root bark is taken as a beverage. A decoction of the leaves and young branches is drunk to treat jaundice. A decoction of the leaves is also given to children to treat rickets.

[0012] Different parts of the plant are used to promote lactation and as an anti-fatigue tonic. A mixture of leaves infused with shea butter is used to rub the skin to treat leprosy.

[0013] In Mali, decoctions of the bark, leaves, and other parts of the plant are used to treat menstrual disorders. The leaves have skin-moisturizing properties, and a warm bath of the leaf decoction can be used for feverish patients, especially children. The leaves are also used to treat hookworms, and the flowers are used to treat tapeworms.

[0014] All of this data comes from an ethnobotanical report written by Dr. Lassina SANOU, Colonel of Water and Forestry, who worked at the Department of Environment and Sustainable Development of the National Centre for Tree Species of Burkina Faso (CNRS) in Ouagadougou (December 2014).

[0015] polysaccharides

[0016] Polyglycosides, polysaccharides, or polysaccharides are arbitrarily defined as high molecular weight polymers produced by the condensation of numerous sugars (d'oses) or sugars (sucres). They participate in various aspects of plant life and even survival. For example, they are responsible for the rigidity of cell walls in higher plants (cellulose, hemicellulose, lignin, etc.), they are forms of energy storage (starch), and due to their hydrophilicity, they can protect tissues against dehydration…

[0017] We are divided into:

[0018] - Homogeneous polysaccharides (condensation of the same type of sugar); and

[0019] - Heteropolysaccharides (hexoses, pentoses, sugar ethers, etc.)

[0020] These two categories can be linear or branched.

[0021] Microbiota and Microbiome

[0022] The skin microbiome is all the microorganisms (bacteria, viruses, fungi, etc.) that exist on and within the skin. Microbiomes even exist in the dermis and adipose tissue, as well as in exocrine glands (Nakatsuji et al. 2013). The microbiome should not be confused with the microbiome, which is a group of genes (the genome of bacteria).

[0023] There are two main types of fungal colonies:

[0024] - Resident flora, composed of symbiotic bacteria, which depend on their host for survival but do not cause harm; and

[0025] - Temporary bacterial colonies, which consist of saprophytic bacteria (harmless) and opportunistic pathogens.

[0026] There are 10¹² bacterial cells (10⁶ / cm²) in the skin. 2 It corresponds to 3,1010 human cells (ratio 33:1 compared to the gastrointestinal system: ratio 25:1); 500 different bacteria; 1 resident or transient flora; and 4 major taxa (Phyla).

[0027] There are many skin / microbiome interactions:

[0028] - Symbiosis: The skin represents an ecological niche and serves as a nutrient reservoir for certain bacteria.

[0029] Bacteria can stimulate the immune defense; the responding skin can form a veritable antimicrobial barrier that inhibits pathogen growth and stimulates the production of antimicrobial molecules, which helps maintain good skin health. However, depending on the specific circumstances, the same bacteria can be symbiotic or pathogenic. Symbionts contribute to physiological inflammation, barrier protection, skin surveillance / tolerance (PAM) and its innate immunity, virulence loss, and TLR (Toll-like receptor) isolation. Symbionts live within biofilms. Sometimes, due to various intrinsic and / or environmental factors, these symbionts become out of control and then transform into pathogens. This leads to an increase in the virulence factors of the microbiome, as well as inflammatory phenomena and activation of the immune system. Symbiotic bacteria can sometimes be eradicated.

[0030] In what is considered normal skin condition, the host's immune system maintains the balance of the microbiota. Immune system deficiencies lead to dysbiosis and specific skin pathologies: atopic dermatitis, acne, and psoriasis. Increased colonization of the skin by *Propionibacterium acnes* (P. acnes) was observed during acne, *Staphylococcus aureus* during atopic dermatitis, Firmicutes (and a decrease in actinomycetes) during psoriasis, and *Malassezia furfur* during seborrheic dermatitis. Summary of the Invention

[0031] The applicant has discovered that extracts of Bombax cuneata, preferably extracts of the calyx of Bombax cuneata, have cosmetic and dermatological properties that have never been described before.

[0032] This invention relates to a polysaccharide-rich extract from the calyx of the cotton tree, preferably from the cotton tree.

[0033] In botany, a flower is a part of a plant, which, when complete, includes the following from the outside in:

[0034] - The calyx, composed of all sepals;

[0035] - The corolla, composed of all the petals;

[0036] - Stamens (l'androcée), namely all the stamens (male parts) that produce pollen; and

[0037] - The gynécée or pistil is formed by all the carpels (female parts).

[0038] In the context of this invention, the term "flower" includes at least the corolla and calyx. A flower can also be complete, that is, it includes the calyx, corolla, stamens, and gynoecium or pistil.

[0039] The extract according to the present invention is preferably an extract of the calyx of the ribbed cotton flower.

[0040] "Extracts rich in polysaccharides" refers to extracts that mainly or substantially contain polysaccharides, meaning that most of the compounds are polysaccharides.

[0041] Therefore, the extract according to the invention advantageously contains at least 15%, more advantageously at least 20%, preferably at least 30%, and more advantageously at least 50% polysaccharides relative to the total weight of the dried extract. Thus, the extract according to the invention advantageously contains 15% to 65%, more advantageously 20% to 65%, preferably 30% to 65%, and more advantageously 50% to 65% polysaccharides relative to the total weight of the dried extract. The percentages are expressed as relative to the total weight of the dried extract (before the addition of any drying medium), determined according to the phenol sulfate method (Dubois method) or the anthrone method-total sugar spectrophotometric method.

[0042] According to the present invention, the apparent molecular weight of the polysaccharide present in the extract is 1500 kDa to 6000 kDa, advantageously 3000 kDa to 6000 kDa (determined by gas chromatography).

[0043] Advantageously, the polysaccharides in the extracts according to the invention comprise monosaccharides and derivatives selected from galactose, rhamnose, galacturonic acid, glucuronic acid, and mixtures thereof. In the context of this invention, galactose refers to D-galactose and L-galactose. Similarly, the term rhamnose refers to D-rhamnose and L-rhamnose.

[0044] Advantageously, the polysaccharide of the extract according to the invention comprises a mixture of galactose, rhamnose, galacturonic acid and glucuronic acid.

[0045] Advantageously, the polysaccharide in the extract according to the invention comprises (in weight percentage relative to the total weight of all present sugars (monosaccharides)):

[0046] -10% to 16% galactose;

[0047] -10% to 17% rhamnose;

[0048] -10% to 17% galacturonic acid; and

[0049] -3% to 7% glucuronic acid.

[0050] The extracts according to the invention are advantageously substantially free of polyphenols. Typically, the extracts according to the invention contain less than 1% by weight, and advantageously less than 0.5% by weight, of polyphenols relative to the total weight of the dried extract.

[0051] In the context of this invention, the above-mentioned polysaccharide-rich extract is advantageously obtained by solid / liquid extraction of the flowers, preferably the calyx, of *Ceiba speciosa* in water. The polysaccharides can then be purified and / or redissolved in a suitable solvent to ensure their physical and microbiological stability, or dried by methods known to those skilled in the art.

[0052] The present invention also relates to a method for preparing a polysaccharide-rich extract from the flowers of *Ceiba speciosa*, preferably from the calyx of *Ceiba speciosa*, comprising at least one solid / liquid extraction step in water and under optimal pH, time and temperature conditions known to those skilled in the art.

[0053] Advantageously, according to the present invention, a method for preparing a polysaccharide-rich extract from the flowers of *Ceiba speciosa*, preferably from the calyx of *Ceiba speciosa*, comprises the following sequential steps:

[0054] a) Crushing the flowers of the ribbed kapok, especially the calyx;

[0055] b) Extract the pulverized flowers, especially the calyxes, in water under optimal time, pH, and temperature conditions;

[0056] c) Separation of the solid and liquid phases by decantation and / or centrifugation and / or precipitation and / or continuous filtration; and

[0057] d) Optionally, use a suitable adjuvant to bleach the liquid phase;

[0058] e) Optionally, dry the extract obtained in step c) or d); and

[0059] f) Stabilize the polysaccharides in the liquid medium in a solvent suitable for preserving the physicochemical properties of the extract and controlling microbial growth.

[0060] Advantageously, according to the present invention, a method for preparing a polysaccharide-rich extract from the flowers of *Ceiba speciosa*, preferably from the calyx of *Ceiba speciosa*, comprises the following sequential steps:

[0061] a) Crushing the flowers of the ribbed kapok, especially the calyx;

[0062] b) Extract the crushed flowers, especially the calyx, from the water;

[0063] c) Separate the solid and liquid phases by decantation and / or centrifugation and / or precipitation and / or continuous filtration;

[0064] d) Optionally, use a suitable adjuvant to bleach the liquid phase;

[0065] e) Optionally, dry the extract obtained in step c) or d); and

[0066] f) Optionally, the extract obtained in step c), d), or e) is made physically and microbiologically stable.

[0067] Step a) of pulverizing plants can be carried out by methods known to those skilled in the art, in particular by using a knife grinder or hammer grinder.

[0068] Extraction step b) is preferably carried out in the presence of water. The percentage of pulverized plant material introduced into the water is advantageously 2% to 10% w / w, preferably 2% to 5% w / w, and more advantageously 2% w / w. This solid / liquid extraction is preferably carried out at a temperature of 20°C to 100°C, particularly 50°C to 90°C, more particularly 70°C to 90°C, and typically 90°C.

[0069] Extraction time is advantageously 30 minutes to 4 hours, particularly 1 hour to 3 hours, and advantageously about 1 hour.

[0070] Step c) of separating the solid and liquid phases is carried out by methods known to those skilled in the art, particularly by decantation, centrifugation and / or continuous filtration and / or purification by precipitation of polysaccharides using a suitable solvent (preferably ethanol or a salt solution). Specifically, during step c), the resulting liquid phase is advantageously purified and concentrated, for example by ultrafiltration and / or sterile filtration. Step c) is carried out until a liquid phase is obtained having desirable transparency and a microbial cleanliness of less than or equal to 100 UFC / g of total bacteria.

[0071] When performing step d) of the bleaching liquid phase, it is carried out by methods known to those skilled in the art, particularly by the addition of adjuvants, such as suitable activated carbon or bleaching clay known to those skilled in the art.

[0072] Advantageously, according to the invention, the extract is obtained by extraction with water, said extract having a very low or essentially no polyphenol content (polyphenols are typically present in trace amounts). The extract according to the invention, preferably obtained by solid / liquid extraction in water, typically has a content of less than 1%, preferably less than 0.5%, by weight relative to the total weight of the dried extract.

[0073] Advantageously, according to the invention, the method according to the invention includes at least one specific filtration and / or purification (step c) and / or bleaching on activated carbon (step d) step, which is capable of removing residual polyphenols if necessary. In particular, the method includes at least one purification step, such as ultrafiltration, which is capable of separating “small” molecules such as polyphenols from “large” molecules (size greater than one million Daltons) such as polysaccharides. Furthermore, advantageously, treatment with activated carbon during bleaching step d) also helps to reduce the level of polyphenols in the extract, so that the polyphenols are advantageously present only in trace amounts at the end of the method according to the invention.

[0074] Advantageously, the polysaccharide-rich extract according to the invention can be stabilized by a drying step e) using methods known to those skilled in the art.

[0075] Drying step e) can be performed, for example, on a carrier of maltodextrin or gum arabic fiber type ( The process is carried out in the presence of the company's CNI. The percentage of dry matter obtained relative to the extract in liquid form is typically 0% to 80% carrier content.

[0076] The extract is preferably obtained by freeze-drying or atomization drying to obtain the final powder.

[0077] Step f) is advantageously performed by partially removing water and replacing it with a solvent to make the product obtained in steps c), d), or e) physically and microbiologically stable. The solvent is advantageously selected from vegetable glycerol, glycols, and mixtures thereof, particularly from vegetable glycerol, plant-derived glycols, and mixtures thereof, more particularly from propanediol or propylène glycol, especially 1,3-propanediol, and vegetable glycerol. Advantageously, the amount of water is reduced in the liquid phase by evaporation to less than 50% by weight relative to the total mass of the extract, preferably less than 20%, and step f) is performed by replacing it with an antibacterial or bactericidal solvent selected from vegetable glycerol, glycols, and mixtures thereof, particularly from vegetable glycerol, plant-derived glycols, and mixtures thereof, more particularly from propanediol or propylène glycol, especially 1,3-propanediol, and vegetable glycerol. Then, the water / solvent ratio will advantageously be 50 / 50 to 0 / 100 (w / w), preferably 30 / 70 (w / w) to 10 / 90 (w / w), and advantageously 20 / 80 (w / w).

[0078] Preferably, for example, the polysaccharide-rich extract according to the present invention can be obtained according to the following method:

[0079] a') Dissolve the crushed flowers in water at a 2% (w / w) concentration;

[0080] b') Extract at 90℃ with stirring for 1 hour;

[0081] c') Purification via a series of filtration steps;

[0082] d') Aseptic filtration; and

[0083] e') The resulting extract is stabilized by evaporating water and adding glycerol to obtain a water / glycerol ratio of 50 / 50 to 0 / 100, advantageously 30 / 70 to 10 / 90.

[0084] In the remainder of the specification, "extract according to the invention" will be considered to refer to the extract as defined above, or to an extract that can be obtained by the method according to the invention as described above. Extracts that can be obtained by the method according to the invention described above have the same composition as the extracts according to the invention as defined above.

[0085] The present invention also relates to a composition comprising a polysaccharide-rich extract from the flowers of *Ceiba speciosa*, preferably from the calyx of *Ceiba speciosa*, according to the invention, and a water / solvent mixture, wherein the water / solvent ratio (v / v) of the water / solvent mixture is 50 / 50 to 0 / 100, advantageously 30 / 70 to 10 / 90, and more advantageously 20 / 80, wherein the solvent is selected from ethylene glycol, vegetable glycerin, and mixtures thereof, preferably from plant-derived ethylene glycol and vegetable glycerin, and more preferably from 1,3-propanediol and vegetable glycerin.

[0086] Advantageously, the composition comprises, by weight, 0.001 to 30%, advantageously 0.001% to 10%, of the extract according to the invention (expressed as the weight of the dried extract relative to the total weight of the composition), and by weight, 50% to 99.999%, advantageously 70% to 99.999%, of a water / solvent mixture relative to the total weight of the composition, wherein the water / solvent ratio is 50 / 50 to 0 / 100, advantageously 30 / 70 to 10 / 90, and more advantageously 20 / 80, wherein the solvent is selected from ethylene glycol, vegetable glycerin, and mixtures thereof, preferably from plant-derived ethylene glycol and vegetable glycerin, and more preferably from 1,3-propanediol and vegetable glycerin. According to this aspect of the invention, the amount of solvent has an effective physical and microbiological stabilizing effect on the composition according to the invention, particularly the extract according to the invention.

[0087] The present invention also relates to a composition comprising, as an active ingredient, a polysaccharide-rich extract from the flowers of *Ceiba speciosa*, preferably from the calyx of *Ceiba speciosa*, according to the present invention, and, where appropriate, suitable excipients. The extract according to the present invention is as defined above in the paragraphs concerning the extract itself and in the paragraphs concerning extracts obtainable by the method according to the present invention.

[0088] The composition is advantageously a cosmetic, pharmaceutical, or dermatological composition. Preferably, the composition is formulated for application via a topical external route.

[0089] Advantageously, the composition according to the invention comprises 0.001 to 10%, typically 0.01 to 5%, of the extract according to the invention relative to the total weight of the composition, expressed as a dried extract.

[0090] The compositions according to the present invention may also contain one or more other active ingredients.

[0091] The compositions according to the invention can be formulated into various formulations suitable for topical application, particularly including creams, emulsions, lotions, ointments, lotions, oils, aqueous or hydroalcoholic or glycol solutions, powders, patches, sprays, shampoos, varnishes or any other topical products.

[0092] Depending on their properties (cosmetic, pharmaceutical, or dermatological), the compositions according to the invention may also contain at least one cosmetically, pharmaceutically, or dermatologically acceptable excipient. In particular, the compositions according to the invention may also contain at least one cosmetically, pharmaceutically, or dermatological adjuvant known to those skilled in the art, said adjuvant being particularly selected from surfactants, thickeners, preservatives, fragrances, dyes, chemical or mineral filters, humectants, and thermal spring water. Those skilled in the art know how to use their common knowledge to formulate compositions according to the invention.

[0093] The optimal dosage and galenal form of the composition according to the invention can be determined based on criteria typically considered when establishing a pharmacological or dermatological treatment suitable for a patient or animal, such as the patient's or animal's age or weight, overall severity of the condition, tolerance to the treatment, observed side effects, and skin type.

[0094] The present invention also relates to extracts or compositions according to the invention for the prevention and / or treatment of diseases or pathological conditions of immature, normal, or mature / aged skin and / or mucous membranes (gingiva, periodontal tissue, genital mucosa) and / or skin appendages (hair and nails), particularly inflammatory reactions, oxidative reactions, diseases associated with pollution-related or unrelated free radical attacks, diseases or pathological conditions associated with microbial attacks, barrier or homeostasis disorders, aging, particularly temporal and / or photoaging, diseases or pathological conditions associated with mechanical and / or thermal invasion of skin and / or mucous membranes and / or skin appendages; more advantageously, for the use of inflammatory or irritative reactions or pathological conditions or barrier disorders or homeostasis disorders of immature, normal, or mature / aged skin, skin appendages and / or mucous membranes (gingiva, periodontal tissue, genital mucosa).

[0095] The present invention also relates to the use of extracts or compositions according to the invention for the prevention and / or treatment of imbalances and / or diseases associated with an imbalance of the microbiota of immature, normal, or mature / aged skin and / or mucous membranes (gingiva, periodontal tissue, genital mucosa) and / or skin appendages and / or their attachments.

[0096] In fact, the extracts according to the present invention possess protective activity against mechanical, microbial, thermal, and free radical attacks on immature, normal, or mature / aged skin and / or mucous membranes (gingiva, periodontal tissue, genital mucosa) and / or skin appendages and / or their attachments. The extracts according to the present invention are used to protect the microbiome, thus combating microbiome imbalance. Furthermore, the extracts according to the present invention can stimulate the skin's immune defense and antioxidant systems.

[0097] This invention also relates to the use of extracts or compositions according to the invention in the preparation of pharmaceutical or dermatological compositions for the prevention and / or treatment of diseases or pathological conditions of immature, normal, or mature / aged skin and / or mucous membranes (gingiva, periodontal tissue, genital mucosa) and / or skin appendages (hair and nails), particularly inflammatory reactions, oxidative reactions, diseases associated with pollution-related or unrelated free radical attacks, diseases or pathological conditions associated with microbial attacks, barrier or homeostasis disorders, aging, particularly temporal and / or photoaging, diseases or pathological conditions associated with mechanical and / or thermal invasion of skin and / or mucous membranes and / or skin appendages; more advantageously in the preparation of pharmaceutical or dermatological compositions for the prevention and / or treatment of inflammatory, irritative reactions or pathological conditions or barrier disorders or homeostasis disorders of immature, normal, or mature / aged skin, skin appendages and / or mucous membranes (gingiva, periodontal tissue, genital mucosa).

[0098] The present invention also relates to the use of extracts or compositions according to the invention in the preparation of pharmaceutical or dermatological compositions for the prevention and / or treatment of imbalances and / or diseases associated with imbalances in the microbiota of immature, normal, or mature / aged skin and / or mucous membranes (gingiva, periodontal tissue, genital mucosa) and / or skin appendages and / or their appendages.

[0099] The present invention further relates to a method for preventing and / or treating diseases or pathological conditions of immature, normal, or mature / aged skin and / or mucous membranes (gingiva, periodontal tissues, genital mucosa) and / or skin appendages (hair and nails), particularly inflammatory reactions, oxidative reactions, diseases associated with pollution-related or unrelated free radical attacks, diseases or pathological conditions associated with microbial attacks, barrier or homeostasis disorders, aging, particularly temporal and / or photoaging, diseases or pathological conditions associated with mechanical and / or thermal invasion of skin and / or mucous membranes and / or skin appendages; more advantageously, a method for treating inflammatory or irritative reactions or pathological conditions or barrier disorders or homeostasis disorders of immature, normal, or mature / aged skin, skin appendages (hair and nails) and / or mucous membranes (gingiva, periodontal tissues, genital mucosa), comprising administering, particularly topically, an effective amount of an extract or composition according to the invention to a subject in need.

[0100] The present invention also relates to a method for preventing and / or treating imbalances and / or diseases associated with an imbalance of the microbiota of immature, normal, or mature / aged skin and / or mucous membranes (gingiva, periodontal tissue, genital mucosa) and / or skin appendages and / or their attachments, comprising administering, in particular, topically, an effective amount of an extract or composition according to the invention to a subject in need.

[0101] In particular, the compositions or extracts according to the invention are intended to prevent and / or treat inflammatory or irritant reactions or pathological conditions or barrier disorders or homeostasis disorders of immature, normal or mature / aged skin, skin appendages (hair and nails) and / or mucous membranes (gingiva, periodontal tissue, genital mucosa).

[0102] Favorably, skin inflammation, irritation, disease or pathological condition, barrier disorder, or homeostasis disorder include: acne, rosacea or lupus erythematosus, vascular diseases, especially erythematous and rosacea, diaper dermatitis, atopic dermatitis, eczema, contact dermatitis, irritant dermatitis, allergic dermatitis, seborrheic dermatitis (cradle cap), sensitive skin, reactive skin, dry skin (xerosis), dehydrated skin, skin redness, skin erythema, aging or photoaging skin, photosensitive skin, pigmented skin (melasma, post-inflammatory hyperpigmentation, etc.), stretch marks, sunburn, chemical, physical (e.g., stress in pregnant women), bacterial, fungal irritants, skin aging, especially photoaging and diseases associated with chemical or atmospheric pollution and / or free radical attacks related to exposure to UV or IR.

[0103] Advantageously, inflammation, irritation, disease or pathological condition of the mucosa, or barrier disorder or homeostasis disorder can cause gingivitis (neonatal gingival sensitivity, hygiene problems, due to smoking, etc.) and external or internal irritation of the male or female genital area.

[0104] Favorably, inflammation, irritation, disease or pathological condition of skin appendages, or barrier disorders or homeostasis disorders can cause brittle nails, fragile nails, fragile hair, brittle hair, or dry hair.

[0105] Favorably, reactions, diseases, or pathological conditions associated with skin microbiome imbalance include atopic dermatitis, eczema, the appearance of unpleasant bromhidrosis, weakened skin barrier, acne, psoriasis, and hidradenitis suppurativa.

[0106] Favorably, reactions, diseases, or pathological conditions associated with imbalances in the skin appendage microbiota include folliculitis, crusted scalp, dandruff, and pruritus.

[0107] Favorably, reactions, diseases, or pathological conditions associated with mucosal microbiota imbalance include pruritus, irritation, candidiasis, and bacterial vaginosis.

[0108] The present invention also relates to the use of extracts or compositions according to the invention for the advantageous treatment and / or prevention in healthy subjects of skin dehydration, skin redness, aging or photoaging skin, photosensitive skin, skin aging, especially photoaging, and diseases associated with chemical or atmospheric pollution and / or with free radical attack related to exposure to UV or IR.

[0109] The present invention also relates to the use of extracts or compositions according to the invention for the advantageous care of skin appendages in healthy subjects, particularly for the treatment and / or prevention of brittle nails, fragile nails, brittle hair, and dry hair.

[0110] The present invention also relates to a method of care for skin and / or skin appendages and / or mucous membranes to advantageously improve condition and / or appearance in healthy subjects, comprising applying a composition or extract according to the invention.

[0111] In particular, the present invention relates to a method for the care of skin and / or skin appendages to advantageously prevent barrier alteration and dehydration in healthy subjects, comprising applying a composition or extract according to the invention to the skin and / or hair and nails.

[0112] In particular, the present invention relates to a method of care for skin and / or skin appendages to advantageously prevent and / or treat in healthy subjects alterations to the skin barrier, skin dehydration, skin redness, aging or photoaging skin, photosensitive skin, skin aging, especially photoaging, diseases associated with mechanical or thermal aggression of the skin and diseases associated with free radical attack related to chemical or atmospheric pollution, and / or diseases associated with exposure to UV or IR, comprising applying a composition or an extract according to the invention.

[0113] The following examples illustrate the invention in a non-limiting manner. Attached Figure Description

[0114] Figure 1 Growth curves for Propionibacterium acnes and Malassezia furfur (see Example 2-Vb.).

[0115] Figure 2 This shows the change in the effect of BCP active ingredients on the bacterial growth of different strains in co-culture over time (see Example 2-Vb.).

[0116] Figure 3 Analysis of the morphology of RHE after hematoxylin / eosin staining (see Example 2-VI-b.).

[0117] Figure 4a and Figure 4b Growth curves for Lactobacillus strains (see Example 2-VII-b.).

[0118] Figure 5 This indicates the analysis of biofilms formed by different strains of the skin microbiome in the presence of the active ingredient BCP (see Example 2-IX). Detailed Implementation

[0119] Example

[0120] Example 1: Preparation of polysaccharide solution from the calyx of Bombax cuneata.

[0121] - Grind the calyx of the Bombax ceiba and then suspend it in water with stirring at a ratio of 2% w / w calyx / water;

[0122] - Extract at 90℃ with stirring for 1 hour;

[0123] - The solution obtained by centrifugation is used to separate the solid residues of the plant;

[0124] - Bleaching and filtration are performed by adding activated carbon;

[0125] - Polysaccharides were concentrated to 15 kDa via ultrafiltration;

[0126] - Add glycerin and evaporate water under vacuum to obtain a final glycerin concentration of 80% w / w.

[0127] Analytical characteristics of the obtained polysaccharide solution

[0128] - Orange viscous solution

[0129] -Dry matter (m / m): 1.24%

[0130] -pH: 6.3

[0131] Ash content: 8.9%

[0132] - Total sugar (determined by anthrone method): 35%

[0133] Solution viscosity (TA, mobile 1, 2 rpm): 2592 Cps

[0134] Analytical characteristics (dry matter) of the obtained polysaccharide:

[0135] - Average molecular weight (determined by gas chromatography): 3711 kDa

[0136] -Composition (determined by gas chromatography): 15% galactose; 17% rhamnose; 17% galacturonic acid; 5% glucuronic acid

[0137] Example 2: Bioactivity

[0138] The potential bioactivity of the extract was investigated through gene expression regulation assays in dermal fibroblasts and melanized reconstructed epidermis. Therefore, the expression of 96 genes important for skin and cosmetic physiology was studied in fibroblasts and melanized reconstructed epidermis using PCR arrays.

[0139] a. Materials and Methods:

[0140] The BCP extract (referred to as BCP extract) of the cottonwood polysaccharide according to Example 1 was added at 0.05% dry matter to the culture medium of normal human dermal fibroblasts (NHDF) or reconstructed melanotic human epidermis.

[0141] After 6 or 24 hours of incubation, the expression of the selected markers was evaluated by quantitative RT-PCR (TaqMan microfluidic card). The change in expression of the studied markers compared with the control was expressed as a relative number (QR, QR>1: increase, QR<1: decrease).

[0142] b. Result:

[0143] The most significant results demonstrating the effect of BCP extract on reconstructing gene expression in the epidermis are shown in Table 1 below.

[0144] Table 1: Changes in gene expression of interest in the epidermis of melanized reconstructed human skin

[0145] QR (relative number compared to control) = 1

[0146] Determine the p-value after the Student test.

[0147]

[0148]

[0149] These results indicate that BCP extracts are of particular interest for the following activities by altering the gene expression of certain markers:

[0150] Lipid synthesis and remodeling in epidermal barrier function

[0151] BCP extract increases the gene expression of two enzymes involved in the synthesis or remodeling of lipids within the stratum corneum: the RAB11A gene of GTPase (Ras-associated protein Rab-11A) and the SMPD1 gene of sphingomyelin phosphodiesterase.

[0152] By increasing the expression of these two genes, the active ingredient BCP can enhance the barrier function and hydration of the epidermis.

[0153] Mechanisms regulating keratinocyte adhesion, migration, proliferation, and differentiation:

[0154] The active ingredient in BCP increases the expression of multiligand proteoglycan-1 (SDC1).

[0155] Stem cell protection:

[0156] Keratin 19, encoded by the KRT19 gene, is an epithelial marker and is considered a marker for epidermal stem cells.

[0157] BCP extract has a protective effect on stem cells by stimulating the expression of KRT19.

[0158] Inhibition of melanin production:

[0159] BCP extract reduces MITF gene expression. Therefore, by decreasing MITF gene expression, BCP extract exhibits melanin production inhibitory activity.

[0160] Table 2 below shows the most important results of BCP extract on gene expression in fibroblasts.

[0161] Table 2: Changes in gene expression of interest in normal human dermal fibroblasts (NHDF)

[0162] QR (relative number compared to control) = 1

[0163] Determine the p-value after the Student test.

[0164]

[0165]

[0166] These results demonstrate the activity of the BCP extract in the following aspects:

[0167] Protection against oxidative stress:

[0168] BCP extract induces the expression of several enzymes involved in antioxidant defense: superoxide dismutases, particularly superoxide dismutase 1 (Cu / ZnSOD) encoded by the SOD1 gene and superoxide dismutase 2 (MnSOD) encoded by the SOD2 gene.

[0169] The active ingredient in BCP also induces the expression of one of the two isoforms of metallothionein 1 (MT1G).

[0170] The active ingredient in BCP also induces the TXNRD1 gene, which encodes subtype 1 of thioredoxin reductase.

[0171] The combined induction of these genes encoding proteins with antioxidant and / or detoxification activities confers protection against UV radiation and heavy metals from sources such as urban pollution.

[0172] Anti-aging effects on the extracellular matrix and skin elasticity:

[0173] The primary function of fibroblasts in the dermis is to produce, degrade, and thus regulate the components of the extracellular matrix (MEC) with which they interact. The MEC is a complex structure formed by a network of collagen fibers, elastin fibers, and structural glycoproteins.

[0174] Elastin (ELN) can be found in MEC proteins regulated by the active ingredient of BCP, as well as fibrin-1 and fibrin-2 encoded by the FBN1 and FBN2 genes, respectively. In addition to inducing the ELN and FBN1 genes, the active ingredient of BCP also induces FBN2, which also constitutes microfilaments.

[0175] While acting on elastic fibers, the active ingredient BCP also acts on collagen fibers because it increases the expression of the α1 subunit (COL3A1) of collagen 3.

[0176] The combined effects of BCP on elastin and collagen fiber components, as well as the expression of MMP1 and pegs, are trending towards influencing skin elasticity, which is particularly noteworthy in the field of anti-aging.

[0177] Furthermore, it has been demonstrated that skin aging is associated with a decrease in fibroblast proliferation. The expression of the age-related proliferation factor Ki-67 is decreased (Ma, C. et al., 2011. Expression of metallothionein-I and II in skinaging and its association with skin proliferation. Br. J Dermatol., 164(3), pp. 479-482). The increased expression of MKI67 by the active ingredient BCP indicates increased cell proliferation, thereby enhancing its anti-aging effect.

[0178] Lamin:

[0179] It has been demonstrated that the expression of lamin B1 (LMNB1) in the skin decreases with age. Therefore, lamin B1 is considered a marker of skin cell aging. In fact, it has been shown that age-related or skin pathological conditions are accompanied by a loss of lamin B1 expression at both the protein and mRNA levels (Dreesen, O. et al., 2013. The contrasting roles of lamin B1 in cellular aging and human disease. Nucleus, 4(4), pp. 283-290).

[0180] Therefore, the increased expression of LMNB1 in dermal fibroblasts indicates that the active ingredient of BCP has the effect of reducing cellular senescence that may be related to age or skin pathology, and on the other hand, it is beneficial to maintain core structure and integrity.

[0181] Autophagy:

[0182] BCP extract induced a decrease in SESN2 expression. SESN2 is specifically involved in the UV response of skin cells. This progressed towards increased mTORC1 complex activity, which is closely associated with decreased autophagy activity and thus related to the regulation of mitophagy activity.

[0183] Skin healing:

[0184] Within adhesion sites, integrins bind to intracellular actin filaments via adhesion proteins. Among these cytoplasmic adhesion proteins, ankle protein-1, encoded by the TLN1 gene overexpressed as an active component of BCP, is particularly likely to be found. It forms the initial bond between integrins and the actin cytoskeleton. The binding of ankle protein to integrins regulates their affinity for MECs, while the binding of ankle protein to actin constitutes the first link in cell contraction mechanisms. These two events are particularly important for cell migration. In fact, this involves the cyclic attachment and disengagement of integrins to MECs, as well as the generation of forces required for translocation of cellular contents (Atherton, P. et al., 2015. Vinculin controls talin engagement with the actomyosin machinery Nat. Commun, Vol. 6, p. 10).

[0185] Increased expression of ankle protein-1 and peg protein demonstrates a beneficial effect on cell migration, particularly during skin healing.

[0186] Insufficient PTGS2 expression in an anti-inflammatory environment:

[0187] Prostaglandin-endoperoxide synthases, including cyclooxygenase-2COX2 or PTGS2, catalyze the biosynthesis of prostaglandins (PGs) from arachidonic acid, which induce inflammatory processes via cytokine secretion and skin vasodilation.

[0188] The reduction in COX2 expression indicates that the active ingredient of BCP can reduce inflammation that can be induced by various stimuli (such as pathogens, UV, ionizing radiation, etc.).

[0189] I. Anti-inflammatory activity

[0190] The anti-inflammatory activity of the cottonwood polysaccharide extract according to Example 1 has been confirmed by PMA-induced inflammation.

[0191] a) Materials and Methods

[0192] Normal human keratinocytes (KHN) were treated at 37°C for 24 hours with 0.01% and / or 0.05% dry matter (MS) of the *Kaempferia galanga* polysaccharide extract according to Example 1 (referred to as BCP extract) or 10⁻⁷ M of dexamethasone or indomethacin (an anti-inflammatory reference molecule). The cells were then treated at 37°C for 16 hours with the addition of 10 μg / ml PMA (phorbol myristate acetate, an inflammation inducer).

[0193] At the end of the treatment, the amounts of IL1α (interleukin 1α), PGE2 (prostaglandin-E2), IL1β (interleukin 1β), TNFα (tumor necrosis factor α), IL6 (interleukin 6), and IL8 (interleukin 8) secreted in the culture supernatant were measured by ELISA.

[0194] The significance of the results was assessed using one-way ANOVA and subsequent Tuckey test.

[0195] b) Results

[0196] As shown in Tables 3 to 8, BCP extract significantly inhibited the production of inflammatory mediators IL1α, IL1β, IL6, IL8, TNFα and PGE2 in keratinocytes under inflammatory conditions.

[0197] These results demonstrate the anti-inflammatory activity of the BCP extract.

[0198] Table 3: Dosage of IL1-α produced by KHN under PMA stress

[0199] $$p<0.01 relative to control; *p<0.05**p<0.01 relative to PMA; ns not significant

[0200] One-way ANOVA and subsequent Tukey test

[0201]

[0202] Table 4: Dosage of IL1β produced by KHN under PMA stress

[0203] $$p<0.01 relative to control; *p<0.05 relative to PMA; ns not significant

[0204] One-way ANOVA and subsequent Tukey test

[0205]

[0206] Table 5: Dosage of IL6 produced by KHN under PMA stress

[0207] $p<0.05 relative to control; *p<0.05***p<0.001 relative to PMA

[0208] One-way ANOVA and subsequent Tukey test

[0209]

[0210] Table 6: Dosage of IL8 produced by KHN under PMA stress

[0211] $$$p<0.001 relative to control; *p<0.05***p<0.001 relative to PMA; ns not significant one-way ANOVA and subsequent Tukey test

[0212]

[0213] Table 7: Dosage of TNFα produced by KHN under PMA stress

[0214] $$$p<0.001 relative to control; *p<0.05***p<0.001 relative to PMA

[0215] One-way ANOVA and subsequent Tukey test

[0216]

[0217]

[0218] Table 8: Dosage of PGE2 produced by KHN under PMA stress

[0219] $$$p<0.001 relative to control; **p<0.01***p<0.001 relative to PMA; ns not significant

[0220] One-way ANOVA and subsequent Tukey test

[0221]

[0222] II. Epidermal healing activity

[0223] The potential activity of the BCP extract (referred to as BCP extract) according to Example 1 on epidermal healing was evaluated by studying keratinocyte migration (the first step in the skin re-epithelialization process).

[0224] a) Materials and Methods

[0225] Scratch test: Damage was performed on a fused normal human keratinocyte (KHN) monolayer pad, followed by treatment with 0.01% and 0.05% dry matter BCP extract or with 100 ng / ml EGF (epidermal growth factor).

[0226] Photographs were taken at the start of treatment and after 5 hours of incubation to measure the rate of KHN development within the lesion. Coverage percentage was evaluated under different conditions through image analysis.

[0227] The significance of the results was assessed using a Student's t-test.

[0228] b) Results

[0229] As demonstrated by the results in Table 9, BCP extract significantly stimulated keratinocyte migration, thus confirming its potential to activate skin healing.

[0230] Table 9: KHN migration *p<0.05**p<0.01***p<0.001 relative to control; t-test

[0231] %recover Changes compared to the control Comparison 33.6±3.1 0 EGF 100ng / ml 62.2±11.1 +85％** BCP 0.01% ms 55.4±14.1 +65％* BCP 0.05% ms 67.0±8.6 +99％***

[0232] III. Activity against innate antimicrobial defenses

[0233] The effect of the polysaccharide extract of Bacillus riberi (referred to as BCP extract) according to Example 1 on the expression of antimicrobial peptide (PAM) and TLR2 in epidermal keratinocytes was evaluated.

[0234] a) Materials and Methods

[0235] Normal human epidermal keratinocytes were treated with BCP extract or 100 ng / ml IL1β (positive reference) for 24 or 48 hours.

[0236] At the end of the treatment, the gene expression of hBD2, hBD3 and TLR2 was analyzed by real-time quantitative RT-PCR.

[0237] In addition, the levels of TLR2 and intracellular hBD2 were measured by ELISA.

[0238] b) Results

[0239] As demonstrated by the results in Tables 10 and 11, the BCP extract significantly stimulated the gene and protein expression of defensins and TLR2; thus demonstrating antimicrobial activity and activation of immune defense.

[0240] Table 10: Gene expression of PAM and TLR2 in keratinocytes

[0241] (QR: Relative Quantity)

[0242] *p<0.05; **p<0.01; ***p<0.001; ns was not significant compared to the control.

[0243] One-way ANOVA and subsequent Dunnett test

[0244]

[0245] Table 11: Production of hBD2 and TLR2 in keratinocytes

[0246] (Intracellular protein expression)

[0247] *p<0.05; **p<0.01; ***p<0.001; ns was not significant compared to the control.

[0248] One-way ANOVA and subsequent Dunnett test

[0249]

[0250] IV. Activity against bacterial adhesion

[0251] The effects of BCP extract on the adhesion of four bacterial strains to the surface of reconstructed human epidermis (RHE) or skin explants were evaluated.

[0252] a) Materials and Methods

[0253] - Studies on the adhesion of Staphylococcus aureus, Staphylococcus epidermidis, and Propionibacterium acnes:

[0254] Human reconstructed epidermis (RHE) was incubated for 15 minutes in the presence of 0.6% and 2% of the *Gossypium riberi* polysaccharide extract according to Example 1 (referred to as BCP extract). Then, after rinsing twice with PBS, *Staphylococcus epidermidis* (ATCC 14990), *Staphylococcus aureus* (ATCC 6538), or *Propionibacterium acnes* (ATCC 6919) bacterial strains were deposited on the RHE surface for 4 hours. After removing unadhered bacteria through four consecutive washes, the RHE was incubated again overnight.

[0255] Bacterial counting was performed by inoculating ground RHE onto a specific agar and then counting the colonies. The results are expressed as UFC / RHE (colony forming units).

[0256] - Adhesion studies of Corynebacterium xerosis:

[0257] Skin explants were incubated for 2 hours in the presence of 0.6% and 2% BCP extracts. Then, a suspension of Corynebacterium xerosis (DSM 20743) was deposited on the surface of the skin explants for 2 hours. After rinsing, the explants were incubated again for 24 hours.

[0258] Bacteria adhering to the surface of the skin explant were recovered by scraping and then inoculated onto agar for UFC / cm 2 The count is represented.

[0259] b) Results

[0260] At both tested concentrations, the BCP extract significantly inhibited the adhesion of Staphylococcus aureus to RHE. At 2%, the BCP extract also inhibited the adhesion of Propionibacterium acnes. (See Table 12)

[0261] BCP extract significantly inhibited the adhesion of dried Corynebacterium tumefaciens to the surface of explants (see Table 13).

[0262] BCP extract promotes the rebalancing of the skin microbiome by limiting the adhesion of pathogens while maintaining the adhesion of symbiotic bacteria.

[0263] Table 12: Bacterial adhesion on RHE

[0264]

[0265] Table 13: Adhesion of *Corynebacterium spp.* on skin explants **p<0.01; ***p<0.001 - one-way ANOVA and subsequent Dunnett test

[0266] <![CDATA[UFC / cm 2 ]]> change Significance Comparison 597±190 BCP 0.6% 272±114 -54％ ** BCP 2% 48±38 -92％ ***

[0267] V. Activity against bacterial growth

[0268] The antimicrobial activity, or conversely, the prebiotic activity, of the cottonwood polysaccharide extract (referred to as BCP extract) according to Example 1 was evaluated by studying the bacterial growth of different strains cultured separately or in co-culture.

[0269] a) Materials and Methods

[0270] Use the following microbial strains that represent the skin microbiome:

[0271] Staphylococcus aureus (ATCC 6538);

[0272] Staphylococcus epidermidis (ATCC 12228);

[0273] Propionibacterium acnes (ATCC 11827);

[0274] Corynebacterium tumefaciens (ATCC 373);

[0275] Malassezia furfur (ATCC 14521);

[0276] Staphylococcus hominis (ATCC 27844).

[0277] - Determination of minimum inhibitory concentration (CMI) and determination of bacterial growth "activator" potential:

[0278] BCP extracts were diluted in microplates in minimum culture media specific to each strain at concentrations of 0.25%, 0.5%, 1%, 2%, 4%, and 8%. Controls were prepared similarly to the samples.

[0279] - Positive growth inhibition control: in diluted medium 5%,

[0280] - A control consisting of diluted growth medium.

[0281] - A control consisting of growth media.

[0282] The strains were added under different conditions and then incubated for 48 hours.

[0283] In cases where disease exhibiting microbial growth occurred after 48 hours, aliquots were collected from the three highest concentrations still showing microbial growth. These aliquots were deposited on separate plates at decimal dilutions to determine the population concentration through limiting dilution.

[0284] After 48 hours of incubation, bacterial growth activation was demonstrated by determining the concentration of each microorganism based on the decimal dilutions performed (by visual reading and simultaneous measurement of absorbance (turbidity) at 620 nm).

[0285] Finally, the results were presented by comparison with bacterial growth obtained in a control group without the product.

[0286] - Determination of the effect on bacterial growth in co-culture:

[0287] Co-cultured strains of Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus hominis, and Propionibacterium acnes were incubated for 48 hours in the presence of 0.25% BCP extract, 0.5% and 1% or Phenoponip 0.5%, and a positive growth inhibition control.

[0288] Remove equal portions of the sample and place them on a specific agar plate for the selected strain. After incubation for 48 hours, count the strains.

[0289] b) Results

[0290] - Determination of minimum inhibitory concentration (CMI) and determination of bacterial growth "activator" potential:

[0291] Inhibition of bacterial growth was observed in *Propionibacterium acnes*, with a measured CMI of 4%. Growth kinetics showed that the BCP extract had antibacterial activity against *Propionibacterium acnes* (see [link to study]). Figure 1 ).

[0292] Similarly, the growth kinetics of Malassezia furfur showed that BCP extract had an antibacterial effect against this yeast (see [link to study]). Figure 1 ).

[0293] - Determination of the effect on bacterial growth in co-culture:

[0294] BCP extract showed trophic effects against Staphylococcus epidermidis, Staphylococcus hominis, and Propionibacterium acnes strains, and tended to inhibit the growth of Staphylococcus aureus (bactericidal effect) (see [link]). Figure 2 ).

[0295] These results demonstrate a protective effect on the balance of the skin microbiome.

[0296] VI. Defense against damage caused by Staphylococcus aureus

[0297] Staphylococcus aureus is frequently detected in patients with atopic dermatitis.

[0298] The amount of Staphylococcus aureus present in these patients is correlated with the severity of the pathological condition and plays an important role in pathophysiology.

[0299] The aim of this study was to reproduce the stress that this bacterium exerts on the epidermis. To this end, Staphylococcus aureus secretions were applied to the RHE surface to study the effects on epidermal differentiation proteins.

[0300] a) Materials and Methods

[0301] 1% of the BCP extract according to Example 1 was applied to the surface of reconstructed human epidermis (RHE) as a 24-hour pretreatment. The RHE was then locally treated by depositing secretions of Staphylococcus aureus (ATCC 33592) (culture medium).

[0302] After a 24-hour incubation, the tissues were morphologically analyzed following hematoxylin / eosin staining, and the expression of barrier function markers was analyzed by immunofluorescence. ImageJ software was used to semi-quantitatively evaluate the expression levels of the proteins of interest.

[0303] b) Results

[0304] Morphological analysis:

[0305] like Figure 3 As shown, Staphylococcus aureus secretions induce moderate changes in RHE morphology: disintegration of the epidermal (basal layer) structure and reduction of keratinocytes in the granular layer.

[0306] Pretreatment with BCP extract preserved the morphology of the epidermis, resulting in better, thicker epidermal tissue and more pronounced production of transparent keratinocytes.

[0307] Expression of barrier function markers:

[0308] As demonstrated by the results in Table 14, Staphylococcus aureus secretions significantly reduced the expression levels of the investigated barrier function markers: corneal desmoprotein, desmosome core protein-1, and filaggrin. Therefore, this model represents the negative impact of Staphylococcus aureus on barrier function, particularly in the pathophysiology of atopic dermatitis.

[0309] BCP extract was able to counteract this reduction in expression.

[0310] Table 14: Expression of barrier markers in RHE

[0311] $p<0.05; $$p<0.01 relative to control; *p<0.05; **p<0.01 relative to secretions - unpaired t-test

[0312] corneal chain protein Desmosome core protein 1 Silk polymers Comparison 100 100 100 Staphylococcus aureus secretions 49.6$ 35.4$$ 42.5$$ Discharge + BCP 1% 84.2* 120.1** 79.4*

[0313] Therefore, BCP extract can protect the skin from Staphylococcus aureus.

[0314] VII. Protective effect against vaginal microbiota

[0315] The prebiotic efficacy of the BCP extract according to Example 1 was evaluated in three lactobacillus strains representing the vagina; in addition, the BCP extract was studied in a vaginal epithelial reconstruction model in which the vaginal epithelium was colonized by lactobacillus strains representing the resident microbiota.

[0316] a) Materials and Methods

[0317] - Studies on the prebiotic effects of Lactobacillus gasseri, Lactobacillus acidophilus, and Lactobacillus rhamnosus:

[0318] Lactobacillus gasseri (ATCC 33323), Lactobacillus acidophilus (LA-14), and Lactobacillus rhamnosus (ATCC 53103) strains were inoculated into depleted media in the presence of 0.25%, 0.5%, 1%, and 2% of the polysaccharide extract of Bacillus riberi according to Example 1 (referred to as BCP extract) or 2% glucose (positive control).

[0319] After 4 hours and 24 hours of incubation, bacterial growth was evaluated by spectrophotometric measurement of DO (indicating bacterial replication rate) at 600 nm and bacterial viability (indicating the number of surviving residual bacteria) in CFU / ml.

[0320] After 24 hours of incubation, the effect on bacterial metabolism was evaluated by measuring the production of lactic acid in the culture supernatant.

[0321] -A study on its protective effect on reconstructing vaginal epithelium:

[0322] A 1% BCP extract was applied to the surface of reconstructed human vaginal epithelium (HVE), which was then colonized by a mixture of Lactobacillus cristataus (DSM 20356) and Lactobacillus gasseri (DSM 20243), mimicking the physiological resident microbiota.

[0323] After 6 hours of incubation, the gene expression of the antimicrobial peptide hBD2 was evaluated by real-time RT-PCR.

[0324] b) Results

[0325] - Research on the prebiotic effects of Lactobacillus gasseri, Lactobacillus acidophilus, and Lactobacillus rhamnosus

[0326] BCP extract showed a positive effect on the growth of *Lactobacillus gasseri* by measuring DO (dosage-dependent) growth rate, starting at 4 hours in a dose-dependent manner (initial prebiotic effect); and a positive effect on the growth of *Lactobacillus acidophilus*, more particularly a prebiotic effect after 24 hours of incubation (stationary period) (see [link to original text]). Figure 4a and Figure 4b ).

[0327] In summary, the BCP extract had a similar or even greater effect on the activity of Lactobacillus than the positive control (glucose), confirming its prebiotic effect.

[0328] Bacterial viability assessment (see Table 15) showed that 2% BCP extract exhibited the greatest prebiotic effect against the three Lactobacillus strains at 4 hours, particularly against *Lactobacillus gasseri* and secondarily against *Lactobacillus rhamnosus*. For *Lactobacillus rhamnosus*, this trend persisted up to 24 hours, while a significant decrease in *Lactobacillus gasseri* viability was observed at 24 hours, which could be explained by culture-related starvation in the depleted medium.

[0329] A 0.25% concentration of BCP extract has a prebiotic effect on Lactobacillus acidophilus.

[0330] Table 15: Bacterial viability of Lactobacillus strains *p<0.05; **p<0.01 - ANOVA test

[0331]

[0332] Lactic acid is a product of primary metabolism in lactobacilli, and its quantification in Lactobacillus acidophilus culture medium is shown in Table 16.

[0333] BCP extract induces an increase in lactic acid production by Lactobacillus acidophilus.

[0334] Table 16: Quantitative analysis of lactic acid produced by Lactobacillus acidophilus *p<0.05; **p<0.01 - ANOVA test

[0335]

[0336] In summary, these results demonstrate the prebiotic effect of BCP extract on Lactobacillus acidophilus. This effect is greater at a concentration of 2%.

[0337] Table 17: Summary of the prebiotic effects of Lactobacillus

[0338] ++ It is more effective than the positive control; high prebiotic effect

[0339] + Equivalent to the effect of a positive control; prebiotic effect

[0340] - No change compared to control; no prebiotic effect.

[0341]

[0342] -A study on its protective effect on reconstructed vaginal epithelium (HVE):

[0343] In the colonized vaginal epithelium (HVE) model, BCP extract significantly increased hBD2 gene expression (see Table 18).

[0344] The results demonstrate the enhanced role of local epithelial defense in protecting the mucosa from the risk of infection.

[0345] Table 18: hBD2 gene expression in HVE colonized by Lactobacillus (QR: relative amount)

[0346]

[0347] VIII. In vitro olfaction test (deodorizing activity)

[0348] An in vitro "smell test" was conducted to evaluate the ability of the BCP extract according to Example 1 to inhibit or alter the odor production of the main bacterial species causing bromhidrosis: Corynebacterium striatum and Staphylococcus epidermidis.

[0349] a) Materials and Methods

[0350] Staphylococcus epidermidis (ATCC 12228) and Corynebacterium striatum (ATCC 6940) strains were used.

[0351] Each of these strains has typical olfactory characteristics, which are the result of the metabolism of sweat components:

[0352] Staphylococcus epidermidis: valeric acid and propionic acid;

[0353] Corynebacterium bandingense: thiol.

[0354] Each bacterial strain was inoculated into reconstituted sweat in the presence of 0.1%, 0.6%, or 2% BCP extract or 0.1% chlorhexidine digluconate (positive control).

[0355] After 6 hours and 24 hours of incubation:

[0356] Bacterial viability is evaluated by bacterial count expressed in CFU / ml.

[0357] A olfactory test is conducted to qualitatively and semi-quantitatively determine olfactory characteristics (odor intensity): immediately after the vial is opened, a trained operator evaluates and scores the odor.

[0358] 1. None (equivalent to the 0.1% chlorhexidine positive control)

[0359] 2. Medium

[0360] 3. Strong (bacterial-specific olfactory characteristic)

[0361] b) Results

[0362] Compared with the negative control, the BCP extract inhibited the odor produced by Staphylococcus epidermidis and Corynebacterium tumefaciens, but did not change the viability of these bacteria (see Table 19).

[0363] These results demonstrate the deodorizing activity.

[0364] Table 19: Bacterial activity and odor in reconstructed sweat **p<0.01 - ANOVA test relative to control

[0365]

[0366] IX. Research on bacterial biofilms

[0367] Biofilms are defined as collections of microbial cells that are associated with living organisms or tissues and “encased” in a polysaccharide matrix.

[0368] Biofilms are one of the most important virulence factors in infectious diseases. They confer resistance to antibiotics, prevent host defense, and increase the virulence of pathogens by promoting the communication system of pathogens (quorum sensing).

[0369] Biofilms in the skin have been described in various pathological conditions or diseases: acne, rosacea, atopic dermatitis. Biofilms can also disrupt healing mechanisms.

[0370] Bacteria in biofilms have an inherent resistance to antibiotics, disinfectants, and the host's defense system; therefore, biofilms promote the persistence of pathogens and their resistance to treatment.

[0371] For example, the involvement of Staphylococcus aureus biofilms is specifically described in the pathophysiology of atopic dermatitis. In fact, Staphylococcus aureus is more likely to exist in atopic skin in the form of biofilms, possibly due to changes in the skin surface that enable adhesion; this biofilm state enhances its resistance to treatment and increases its virulence, leading to the induction of chronic inflammation and pruritus, resulting in a vicious cycle in the pathogenesis of atopic dermatitis.

[0372] The effect of the BCP extract according to the present invention on the biofilm formation ability of bacteria in the skin microbiome was evaluated.

[0373] a) Materials and Methods

[0374] - Microbial strains tested:

[0375] - Staphylococcus epidermidis MFP04: A healthy human skin strain (LMSM strain group) characterized by metabolic analysis, ARN16S, and genome sequencing. This is a strain described as symbiotic.

[0376] - Staphylococcus aureus MFP03: A healthy human skin strain (LMSM strain group) characterized by metabolic analysis, ARN16S, and genome sequencing. It is a symbiotic strain, an "opportunistic" pathogen that can cause certain skin infections and is widely described as being associated with the pathophysiology of atopic dermatitis.

[0377] - Corynebacterium tumefaciens CIP 100653T / ATCC373: A strain from an international database. It is a symbiotic strain associated with the production of bromhidrosis (body odor).

[0378] - Propionibacterium acnes HL045PA1 / HM-516 ribonucleic acid type 4 (RT4): A “acne” strain characterized by Fitz-Gibbon et al., obtained from an international database. It is a pathogenic strain involved in the pathogenesis of acne.

[0379] - Propionibacterium acnes HL110PA3 / HM-554 ribonucleic acid type 6 (RT6): A “non-acne” strain characterized by Fitz-Gibbon et al., obtained from an international database. It is a symbiotic strain.

[0380] - Micrococcus luteus 0116: A healthy human skin strain (LMSM strain group) characterized by metabolic analysis, ARN16S, and genome sequencing. It is a symbiotic strain.

[0381] - Preliminary study on the effects of BCP active ingredients on bacterial growth kinetics:

[0382] A control study was conducted to verify that the active ingredient of BCP had no effect on the growth of the six strains studied.

[0383] At the start of incubation, add the active ingredient diluted to 1 / 50 to the medium. Depending on the strain, monitor bacterial growth in the microplates at 24, 48, or 72 hours. Continuously measure absorbance using a microplate reader / incubator.

[0384] - Investigating the role of BCP active ingredients in biofilm formation using crystal violet technology:

[0385] Depending on the strain, after culturing for 24, 48, or 72 hours in the presence of the active ingredient, the formation of bacterial biofilms was studied in multi-well plates (96) using crystal violet staining.

[0386] Results are expressed as a percentage of biofilm formation in the control medium (without active ingredients). Statistical differences were established using the Mann-Whitney test.

[0387] b) Results

[0388] The BCP extract of kapok polysaccharide according to the present invention did not significantly alter the bacterial growth kinetics under the test conditions.

[0389] Studies on biofilm formation by different bacterial strains have revealed the following effects of kapok polysaccharide extract (see...) Figure 5 ):

[0390] - Significantly inhibits Staphylococcus aureus biofilm formation;

[0391] -Induces the formation of Staphylococcus epidermidis biofilm;

[0392] - No significant regulation of biofilm formation of Micrococcus luteus, Corynebacterium sicca, or Propionibacterium acnes RT6 (non-acne strain);

[0393] - Inhibits biofilm formation in Propionibacterium acnes RT4 (acne strain).

[0394] Advantageously, the BCP extract of kapok polysaccharide inhibits the biofilm formation ability of pathogenic bacterial strains (especially Staphylococcus aureus and Propionibacterium acnes RT6), while maintaining or even promoting the biofilm formation ability of symbiotic bacteria.

[0395] These results suggest that the active ingredients play a role in maintaining the balance of the microbiome, particularly beneficial for pathological conditions such as atopic dermatitis or acne.

Claims

1. A type of kapok derived from the ribbed kapok tree Bombax costatum A polysaccharide-rich extract of a flower, comprising at least 15% polysaccharide relative to the total weight of a dried extract, the polysaccharide having an apparent molecular weight of 1,500 to 6,000 kilodaltons, the polysaccharide comprising 10 to 16% galactose, 10 to 17% rhamnose, 10 to 17% galacturonic acid and 3 to 7% glucuronic acid, wherein the extract comprises less than 1% polyphenols relative to the total weight of the dried extract, and wherein the extract is obtained by a preparation method comprising at least one solid / liquid extraction step in water; The preparation method, which includes at least one solid / liquid extraction step in water, comprises the following steps: The flowers of the kapok tree are crushed, the crushed flowers are extracted in water, the solid and liquid phases are separated by decantation and / or centrifugation and / or precipitation and / or continuous filtration, and the resulting liquid phase is purified and concentrated by ultrafiltration and / or sterile filtration.

2. The extract according to claim 1, comprising at least 30% by weight of polysaccharides relative to the total weight of the dried extract.

3. The extract according to claim 1, wherein the extract comprises 20% to 65% by weight of polysaccharides relative to the total weight of the dried extract.

4. A method for preparing a polysaccharide-rich extract from the flowers of *Ceiba speciosa*, comprising at least one solid / liquid extraction step in water, characterized in that... It includes the following sequential steps: a) Crush the flowers of the Bombax ceiba; b) Extract the crushed flowers from water; c) Separate the solid and liquid phases by decantation and / or centrifugation and / or precipitation and / or continuous filtration, and purify and concentrate the resulting liquid phase by ultrafiltration and / or sterile filtration; d) Optionally, use a suitable adjuvant to bleach the liquid phase; e) Optionally, dry the extract obtained in step c) or d); and f) Optionally, the extract obtained in step c), d), or e) is made physically and microbiologically stable.

5. A composition comprising, as an active ingredient, a polysaccharide-rich extract from the flowers of *Ceiba speciosa* according to any one of claims 1 to 3 or an extract obtained by the method according to claim 4, and a suitable excipient.

6. Use of the extract according to any one of claims 1 to 3 or the composition according to claim 5 in the preparation of a composition for the prevention and / or treatment of bromhidrosis, acne and / or bacterial vaginosis.

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