Pharmaceutical or cosmetic composition intended to improve cellulite and varicose veins characterized in that it comprises an association of theanine and polysaccharides derived from matcha.

Abstract

Pharmaceutical or cosmetic composition intended to improve the appearance of cellulite and reduce varicose veins, characterized in that it comprises an association of theanine and polysaccharides extracted from Camellia sinensis, and at least 2 metabolites produced by a Bacillus subtilis bacterium such as Bacillus subtilis Gozen GZN-08.

Description

Title of the invention: Pharmaceutical or cosmetic composition intended to improve cellulite and varicose veins characterized in that it comprises an association of theanine and polysaccharides derived from matcha.

[0001] The present invention relates to a pharmaceutical or cosmetic composition intended to improve the appearance of cellulite and to reduce varicose veins, characterized in that said composition comprises an association of theanine and polysaccharides extracted from the leaves of matcha Camellia sinensis, and at least 2 metabolites produced by the bacterium Bacillus subtilis Gozen GZN-08 from the group: gallic acid, lobelin, biotin, catechin, pyridoxine, syringic acid, butyrate, propionate, zinc, linoleic acid, saikosaponin, acacetin, saponarin, butyric acid.

[0002] Cellulite is known to be a cosmetic problem called "gynoid lipodystrophy (GLD)" affecting 85 to 98% of post-pubertal women. Cellulite is an alteration in the topography of the skin that occurs in the pelvic region, lower limbs, and abdomen. It is characterized by a dimpled or "orange peel" appearance. It is a complex problem involving the microcirculatory and lymphatic systems, as well as the extracellular matrix and the presence of excess subcutaneous fat in the dermis.Indeed, it is characterized by altered adipocytes associated with lymphatic stasis, decreased blood flow, and increased protein in the interstitial fluid, leading to impaired microcirculation. There is also decreased collagen production associated with follicular hyperkeratosis, which causes an orange peel appearance, and altered subcutaneous tissue in the deep dermis, giving the skin a granular texture. Several factors predispose women to GLD: genetics (sex, race, adipose tissue distribution), hormones (insulin, estrogen, prolactin), vascular factors, and poor lifestyle habits (sedentary behavior, diet). Impaired microcirculation can also lead to varicose veins. Varicose veins are an abnormal dilation of a vein and are visible under the skin in the lower limbs, creating an unsightly appearance.To reduce cellulite, that is, to decrease the tension on connective tissue fibers, it is necessary to decrease the volume of adipocytes (fat cells). This involves reducing their triglyceride content and thus stimulating lipolysis or reducing lipogenesis. Lipogenesis is a metabolic pathway that esterifies fatty acids into triglycerides, where they are stored in adipocytes. Triglycerides are produced in the liver, fat cells, and intestinal cells. Lipolysis is the breakdown of triglycerides. stored in adipocytes, that is, their breakdown into non-esterified fatty acids and glycerol. The molecules thus released into the bloodstream can be used by other tissues as an energy substrate (fatty acids) or serve as gluconeogenic precursors (glycerol). One current treatment for cellulite uses plant essences (EP2813207) but requires several essences for it to be effective. Another treatment (W02011152692) using a natural active ingredient, fucoxanthin, is used against cellulite. This treatment activates UCP-1 expression in white fat, which causes apoptosis of fat cells, thus reducing cellulite. However, there are other mechanisms that can reduce the size of adipocytes, such as an increase in intracellular cAMP, a reduction in lipogenesis, or an increase in lipolysis.This treatment is therefore only partially effective in increasing lipolysis. Furthermore, it does not address the reduction of varicose veins or the improvement of microcirculation.

[0003] It was by studying the properties of green tea metabolites that the applicant highlighted that problems related to an increase in the size of adipocytes, an alteration of microcirculation as well as the appearance of varicose veins and an alteration of the extracellular matrix can be improved by a treatment composed of these metabolites.

[0004] To this end, the applicant has developed a pharmaceutical or cosmetic composition intended for topical or oral use comprising a combination of theanine and polysaccharides extracted from matcha leaves Camellia sinensis, combined with at least 2 metabolites, typically produced by the bacterium Bacillus subtilis Gozen GZN-08. These metabolites can be prepared by the fermentation of Bacillus subtilis Gozen GZN-08.

[0005] SUMMARY

[0006] In a first aspect the invention relates to a pharmaceutical or cosmetic composition intended to improve the appearance of cellulite and to reduce varicose veins characterized in that it comprises an association of bioactives including theanine and polysaccharides extracted from the leaves of matcha Camellia sinensis, and at least 2 metabolites produced by a Bacillus subtilis bacterium chosen from the group: gallic acid, lobelin, biotin, catechin, pyridoxine, syringic acid, butyrate, propionate, zinc, linoleic acid, saikosaponin, acacetin, saponarin.

[0007] In one embodiment, the pharmaceutical composition according to the invention comprises zinc at a concentration of at least 0.5% and preferably at least 2% by weight relative to the total weight of the bioactives in said composition and polysaccharides at a concentration of at least 0.5% and preferably at least 5% by weight relative to the total weight of the bioactives in said composition.

[0008] In a second aspect, the invention relates to a manufacturing process for producing the pharmaceutical or cosmetic composition according to the invention. The process comprises a step of natural fermentation of matcha leaves Camellia sinensis in open air, followed by controlled fermentation in the presence of Bacillus subtilis, typically Bacillus subtilis Gozen GZN-08, at a temperature between 25 and 30°C.

[0009] The invention also refers to a pharmaceutical or cosmetic composition that can be obtained by the process of the invention.

[0010] The pharmaceutical or cosmetic composition according to any of the above aspects and embodiments may be in the form of an oil-in-water (O / W) or water-in-oil (W / O) emulsion, a multiple emulsion (O / W / O, W / O / O), a microemulsion, a twin-phase emulsion, a PIT emulsion, a nanoemulsion, a pseudoemulsion, an aqueous gel, a fatty gel, a hydroalcoholic gel, a fatty phase, a suspension, a liquid or solid soap, a foaming or non-foaming solution, a powder, a lyophilized gel or emulsion, preferably in the form of creams, liquids, pastes, lotions, emulsions, oils, gels, syrups, solids, powders, masks, sticks, tablets, sprays, aerosols, capsules, or syrup.

[0011] In a third aspect, the invention relates to the pharmaceutical composition of the invention, for its use in

[0012] - the reduction of fat accumulation in adipose and intestinal tissue by inhibiting the intestinal lipase enzyme and the phosphodiesterase enzyme; and / or

[0013] - to reduce the risk of developing varicose veins, to improve circulation blood and / or to improve microcirculation in the legs.

[0014] In a fourth aspect, the invention relates to the non-therapeutic use of a cosmetic composition according to the invention to tone the skin of the thighs and arms, to thicken the skin of the thighs and arms, to modulate the production of hyaluronic acid by skin cells, to improve the appearance of the skin and increase the elasticity of the skin, or to modulate the thickening of the skin's collagen fibers. DETAILED DESCRIPTION

[0015] The applicant describes below a slow fermentation process for extracts from matcha leaves (Camellia sinensis) and Bacillus subtilis, typically Bacillus subtilis Gozen GZN-08, in several stages. The matcha leaves (Camellia sinensis) are first collected and stored in open air for at least 72 hours to undergo a first fermentation stage. The bacterium Bacillus subtilis, typically Bacillus subtilis Gozen GZN-08, is then inoculated in the presence of the Camellia sinensis plant, which is stored at high density in an environment humid conditions are maintained at a temperature between 20°C and 30°C. This step lasts between 2 and 36 hours and activates the production of metabolites by bacteria, at least one of which is of the genus Bacillus, preferably Bacillus subtilis, and even more preferably Bacillus subtilis Gozen GZN-08. In one embodiment, the Bacillus subtilis bacterium is Bacillus subtilis Gozen GZN-08.

[0016] Indeed, the applicant has discovered that the Bacillus subtilis Gozen GZN-08 strain produces metabolites that can improve microcirculation and inhibit the activity of phosphodiesterase and lipase enzymes. The fermentation step therefore primarily allows for the fermentation of this bacterium of interest. The fermentation product can be extracted and filtered to select the molecules of interest. Extraction can be carried out by aqueous extraction, alcoholic extraction, extraction with another type of solvent, distillation, or extraction with a supercritical fluid. Filtration can be carried out by a liquid chromatography step or by membrane filtration.The object of the present invention is therefore described as a pharmaceutical composition comprising an association of theanine and polysaccharides extracted from the leaves of matcha Camellia sinensis, and at least 2 metabolites, typically produced by the bacterium Bacillus subtilis Gozen GZN-08, selected from the group: gallic acid, lobelin, biotin, catechin, pyridoxine, syringic acid, butyrate, propionate, zinc, linoleic acid, saikosaponin, acacetin, saponarin, butyric acid.

[0017] The use of the composition relates to the reduction of fat accumulation in adipose and intestinal tissue. Lipolysis, the process of breaking down triglycerides into fatty acids, which reduces fat accumulation, is triggered by the hormone-sensitive lipase (HSL) enzyme, which hydrolyzes triglycerides stored in adipocytes and releases fatty acids. The HSL enzyme is activated by cyclic adenosine monophosphate (cAMP). The present invention stimulates lipolysis by inhibiting the action of phosphodiesterase, an enzyme that degrades cAMP, thereby inducing its increase within the adipocyte. In addition, the present invention activates AMPK. This activation inhibits adipogenesis and lipogenesis, two mechanisms of triglyceride formation in adipocytes; while activating lipolysis by regulating the gene expression of various enzymes and transcription factors such as PPARgamma, TNF-alpha or HSL.

[0018] Furthermore, pancreatic lipase is a pancreatic enzyme secreted by the pancreas and released into the small intestine. This enzyme plays a role in lipolysis, as it breaks down triglycerides into fatty acids, which are then absorbed by the small intestine. Inhibiting it reduces fat breakdown and accumulation.

[0019] Polyphenols present in green tea, such as epigallocatechin gallate (EGCG), inhibit pancreatic lipase; however, EGCG has low stability in the intestine. Indeed, EGCG is stable at low temperatures, but at high temperatures (above 25°C), it undergoes polymerization and oxidation. However, mixing EGCG with metabolites produced by Bacillus subtilis, such as Bacillus subtilis Gozen GZN-08, increases its stability. The present invention therefore makes it possible to inhibit the activity of pancreatic lipase by increasing the stability of EGCG. By inhibiting this enzyme, the composition thus inhibits the accumulation of fat in intestinal tissue. The present invention therefore relates to a pharmaceutical composition for use in reducing the accumulation of fat in adipose and intestinal tissue.

[0020] As we age, the skin on our arms and thighs loses its firmness and sags. This phenomenon is visible on the face, but also on the inner arms and thighs. Skin laxity can also occur after significant weight loss. The skin stretches to adapt to increased body mass. Over time, however, this can damage the skin's collagen and elastin, leading to tissue laxity. The present invention activates the gene expression of collagen and elastin by fibroblasts (skin cells specialized in the synthesis of the extracellular matrix that gives skin its shape and structure).

[0021] Hyaluronic acid is an important component of the skin as it helps regulate skin hydration and firmness. The present invention stimulates the production of hyaluronic acid by dermal cells. The composition therefore relates to use in modulating the production of hyaluronic acid by skin cells.

[0022] The use of the composition also relates to improving the appearance and elasticity of the skin. Collagen is an important component of the extracellular matrix, providing strength and flexibility to the skin. Several factors are responsible for its degradation: secretions of matrix metalloproteinases (MMPs) are endopeptidases that cause the proteolytic cleavage of collagen, as well as collagenase and elastase enzymes, which degrade collagen. The present invention increases the thickness of collagen fibers in the skin, thereby increasing skin thickness and elasticity.

[0023] The use of the composition also relates to the improvement of microcirculation. Microcirculation refers to the blood flow in the smallest vessels of the circulatory system, which includes arterioles, venules, and capillaries disseminated throughout all tissues, forming a network located between the arterial and venous systems. Alterations in microcirculation cause changes in skin temperature and color. It is therefore important to improve the Microcirculation is impaired in the case of cellulite. Inflammation induces altered vasomotor function and reduced capillary perfusion, resulting in impaired microcirculation. Thromboxane A2 induces platelet aggregation, which reduces blood flow and circulation. The composition according to the invention (hereinafter referred to as BIOMATCH A) inhibits the activity of thromboxane A2, thereby improving microcirculation. The metabolites present in BIOMATCH A inhibit inflammation, thus improving microcirculation. The composition according to the present invention therefore reduces the risk of developing varicose veins by promoting smooth blood flow and improving microcirculation in the legs.

[0024] The composition according to the present invention is characterized in that it is in the form of an aqueous gel, a greasy gel, a hydro-alcoholic gel, a greasy phase, a suspension, a foaming or non-foaming solution, a powder, an oil-in-water (O / W) or water-in-oil (W / O) emulsion, a multiple emulsion (oil-in-water-in-oil O / W / O, water-in-oil-in-water W / O / W), a microemulsion, a twin-phase emulsion, a PIT emulsion (emulsion obtained by phase inversion at temperature), a nanoemulsion, a pseudo-emulsion (stable dispersion of two immiscible phases by means of a gelling agent), a freeze-dried gel or emulsion, and for its use, and which may be presented, without limitation, in creams, inhalations, liquids, ready-to-use drinks, pastes, lotions, emulsions, oils, tablets, gels, syrups, solids, powders, masks, sprays, aerosols, capsules, stick.The formulation was chosen so that the active ingredient would integrate as effectively as possible and have the best possible effect. EXAMPLES

[0025] Example 1: Inhibition of the phosphodiesterase enzyme.

[0026] An in-vitro study was carried out to measure the ability of BIOMATCHA to inhibit the phosphodiesterase enzyme.

[0027] The phosphodiesterase enzyme PDE5A1 was isolated and incubated with a placebo (water) or BIOMATCHA. The enzyme's activity was then measured by assessing its ability to cleave cyclic guanosine monophosphate (cGMP). Incubation of the phosphodiesterase enzyme PDE5A1 with BIOMATCHA induced a 38% inhibition of its enzymatic activity. The phosphodiesterase enzyme degrades cAMP, which is essential in the hydrolysis of triglycerides in adipocytes.

[0028] Example 2: Improvement of skin appearance

[0029] A double-blind clinical study was conducted to evaluate the effects of applying a formulation containing BIOMATCHA to the skin on the appearance of the skin.

[0030] 42 participants with mild or moderate cellulite were divided by The participants were randomly assigned to two groups of 21 people. The first group (placebo) applied a cream containing a placebo (water), while the second group applied a cream containing 2% biomatcha for four weeks. The level of improvement in skin appearance, varicose veins, and skin tone was assessed by a physician by comparing before-and-after photos.

[0031] The results are presented in Tables 1 and 2 below:

[0032] [Tables 1] Excellent Very good Good Skin appearance PLACEBO 0 1 (4.8%) 4 (19%) with BIOMAT CHA 7 (33.3%) 4 (19%) 8 (38.1%) Skin tone PLACEBO 0 (0%) 1 (4.8%) 6 (28.6%) with BIOMAT CHA 6 (28.6%) 5 (23.8%) 7 (33.3%) Reduction of variations PLACEBO 1 (4.8%) 0 (0%) 7 (33.3%) with BIOMAT CHA 3 (14.3%) 5 (23.8%) 9 (42.9%)

[0033] [Tables2] Slight improvement No improvement PLACEBO appearance 9 (42.9%) 7 (33.3%) BIOMAT CHA 2 (9.5%) 0 (0%) PLACEBO skin tone 11 (52.4%) 3 (14.3%) BIOMAT CHA 3 (14.3%) 0 (0%) PLACEBO reduction 10 (47.6%) 3 (14.3%) BIOMAT CHA 2 (9.5%) 2 (9.5%)

[0034] The results show that the application to the skin of a formulation containing BIOMATCHA results in an improvement in the appearance of the skin and varicose veins.

[0035] Example 3: Modulation of collagen fiber thickness

[0036] A double-blind clinical study was conducted to evaluate the effect of ingesting a food supplement containing BIOMATCHA on the thickness of collagen fibers.

[0037] 29 participants with mild or moderate cellulite were divided by Randomization into 2 groups of 14 and 15 people. The first group (PLACEBO) took a placebo tablet once a day, while the second group took a 150 mg BIOMATCHA tablet once a day for 8 weeks.

[0038] The thickness of the collagen fibers was measured at the beginning and end of the study by ultrasound.

[0039] Consumption of BIOMATCHA induced a significant increase in collagen fiber thickness of 21.8%, while consumption of the placebo did not demonstrate a significant effect.

[0040] These results show that BIOMATCHA has an effect in modulating the thickening of skin collagen fibers.

[0041] Example 4: Modulation of hyaluronic acid production

[0042] An in vitro study was conducted to evaluate the effect of BIOMATCHA on hyaluronic acid production. Human keratinocytes were incubated with or without 1 pM of BIOMATCHA for 48 hours. The release of hyaluronic acid from the keratinocytes was then measured. Incubation with BIOMATCHA increased hyaluronic acid production by 37.5%.

[0043] These results show that BIOMATCHA modulates the production of hyaluronic acid by skin cells.

[0044] Example 5: O / W gel emulsion for the body

[0045] -Carbopol981............................................0.6 g

[0046] - Ethyl alcohol.........................................15 g

[0047] - Volatile silicone oil...............................3 g

[0048] - Purcellin oil........................................3 g

[0049] -BIOMATCHA............................................2 g

[0050] - Perfume...................................................0.4 g

[0051] - Triethanolamine............................................0.2 g

[0052] - Preservatives..........................................0.3 g

[0053] - Demineralized water qsp...................................100 g

[0054] This gel emulsion, applied once a day, helps to reduce the appearance of cellulite.

[0055] Example 6: Dietary supplement to modulate hyaluronic acid production

[0056] - BIOMATCHA...................................................150 mg

[0057]

[0058] - Maltitol, microcrystalline cellulose, silicon dioxide, stearate calcium...................................................150 mg This dietary supplement should be taken twice a day. Users have noticed an improvement in skin quality after 4 weeks.

Claims

Demands

1. A pharmaceutical or cosmetic composition intended to improve the appearance of cellulite and reduce varicose veins comprising a combination of bioactives including theanine and polysaccharides extracted from matcha leaves Camellia sinensis, and metabolites produced by a Bacillus subtilis bacterium selected from the group: gallic acid, lobelin, biotin, catechin, pyridoxine, syringic acid, butyrate, propionate, zinc, linoleic acid, saikosaponin, acacetin, saponarin, said metabolites being derived from the fermentation of matcha leaves Camellia sinensis by a Bacillus subtilis bacterium; characterized in that it comprises zinc at a concentration of at least 0.5% and preferably at least 2% by weight relative to the total weight of bioactives in said composition and polysaccharides at a concentration of at least 0.5% and preferably at least 5%.

2. A manufacturing process for producing the pharmaceutical or cosmetic composition according to claim 1, characterized in that it comprises a step of natural fermentation of Camellia sinensis matcha leaves in the open air, followed by controlled fermentation in the presence of Bacillus subtilis at a temperature between 25 and 30°C, followed by a filtration step by liquid chromatography.

3. Pharmaceutical or cosmetic composition capable of being obtained by the process according to claim 2.

4. Pharmaceutical composition according to claim 1, or claim 3, for its use in reducing fat accumulation in adipose and intestinal tissue.

5. Pharmaceutical or cosmetic composition according to claim 1, or claim 3, characterized in that it is in the form of an oil-in-water (O / W) or water-in-oil (W / O) emulsion, a multiple emulsion (O / W / O, W / O / O), a microemulsion, a twin-phase emulsion, a PIT emulsion, a nanoemulsion, a pseudoemulsion, an aqueous gel, a fatty gel, a hydroalcoholic gel, a fatty phase, a suspension, a liquid or solid soap, a foaming or non-foaming solution, a powder, a gel, or a lyophilized emulsion, preferably in the form of creams, liquids, pastes, lotions, emulsions, oils, gels, syrups, solids, powders, masks, sticks, tablets, sprays, aerosols, capsules, or syrup.

6. Pharmaceutical composition according to claim 1, 3 or 4, for its use in reducing the risk of developing varicose veins.

7. Non-therapeutic use of a cosmetic composition according to claims 1 or 3, to tone the skin of the thighs and arms, to thicken the skin of the thighs and arms, to modulate the production of hyaluronic acid by skin cells, to improve the appearance of the skin and increase skin elasticity, or to modulate the thickening of skin collagen fibers.