Cosmetic composition containing extract of Pseudoalteromonas paragorgicola

Abstract

The present invention relates to the cosmetic use of an extract of Pseudoalteromonas paragorgicola as a slimming agent, more particularly as an anti-hyperplastic agent. The present invention also relates to the use of an extract of Pseudoalteromonas paragorgicola in a slimming composition.

Description

[Technical Field]

[0001] The present invention relates to the field of cosmetic skin treatment. The present invention relates to a cosmetic composition comprising an extract of Pseudoalteromonas paragorgicola. [Background technology]

[0002] The skin is an organ with a surface coating that covers the entire surface of the body and represents a relatively effective protective barrier against external aggressions. The skin is often described as the heaviest organ in the human body. The skin also plays an active role in many biological and biochemical processes.

[0003] Skin is composed of three superimposed layers or tissues, from most superficial to deepest, including the epidermis, dermis and subcutaneous tissue.

[0004] This latter, also known as adipose tissue, accounts for 15-20% of body weight in an average-weight individual. The subcutaneous tissue is attached to the lower dermis by extensions of collagen fibers. The thickness of the subcutaneous tissue varies with age, body position, and gender, and is largely responsible for shaping the body's curves.

[0005] The subcutaneous tissue is a vascularized adipose tissue containing adipocytes that absorb and store fat as triglycerides. It also functions as a storage organ where fat can be mobilized and converted into energy during prolonged activity. It also acts as a protective organ against mechanical shocks and thermal fluctuations to maintain homeostasis. The subcutaneous tissue controls lipid mobilization through two major metabolic activities: lipogenesis (synthesis of fatty acids (FAs) and triglycerides, and storage of surplus energy) and lipolysis (hydrolysis of triglycerides when energy is needed).

[0006] Adipose tissue is not only a storage organ whose overdevelopment is responsible for obesity and associated metabolic disorders, such as type II diabetes, but also a true endocrine organ. This function was assigned to it more than 20 years ago with the discovery of leptin, a hormone secreted by adipose tissue in response to food intake that suppresses appetite by regulating neural circuits located in the brain.

[0007] Adipose tissue is a major organ in maintaining the energy homeostasis of organisms due to its ability to store and release fatty acids and its secretory function. Endothelial cells in the vascular network of adipose tissue form a physical barrier called the endothelium between the blood compartment and adipocytes. Endothelial cells in the vascular network of adipose tissue play a major role in controlling the exchange of nutrients, hormones, oxygen, and immune and inflammatory cells. These exchanges and their regulation are important and specific to the metabolic state of adipose tissue.

[0008] The role of the endothelium in the biology of adipose tissue has evolved considerably from its historical role as an inert layer of cells acting as an internal coating for the poorly developed circulatory system in adipose tissue mass. Current data indicate that the vascular network is highly prevalent in adipose tissue and plays a key role in the development and function of adipose tissue. In addition, the marked heterogeneity of the endothelial layer relative to functional heterogeneity in the context of specific organs such as the heart, liver, and adipose tissue, due to differences in the location of the conducting vessels (arteries, capillaries, and veins), is now well recognized.

[0009] This diversity of the endothelium is important for its specialized roles, including permeability, leukocyte trafficking, and hemostasis, as well as the transport of oxygen, fluid, and nutrients from the circulation to target tissues. The concept of endothelial fatty acid (FA) transport has existed for many years, but it has evolved from a passive diffusive movement to an active, highly regulated, tissue-specific process involving multiple complex signaling pathways. (Hagberg, C.E. et al., Vascular endothelial growth factor B controls endothelial fatty acid uptake. Nature, 2010. 464(7290):917-21).

[0010] FAs, naturally stored in the form of triglycerides in adipocytes, must be channeled through the microvascular endothelium according to mechanisms that are not yet fully understood. Transport of FAs across non-fenestrated microvascular endothelium involves the membrane transporter FATP (fatty acid transport protein) and the cytosolic transporter FABP (fatty acid binding protein). In the endothelium of white adipose tissue (WAT), other factors, including PPARG (peroxisome proliferator-activated receptor gamma), have been proposed to play a role.

[0011] To analyze FA transport in human adipose tissue (hATEC) endothelial cells, Anais Briot and her team measured the ability of freshly isolated primary adipose tissue endothelial cells to absorb fluorescent FA (Bodipy FL C16) in vitro. At 37°C, the cells efficiently take up fluorescent FA. In the presence of the PPARG agonist rosiglitazone at 37°C, an increase in fluorescent FA absorption was observed, demonstrating that treatment with rosiglitazone promotes FA uptake in human adipose tissue endothelial cells (Briot, A. et al., Senescence Alters PPARgamma (Peroxisome Proliferator-Activated Receptor Gamma)-Dependent Fatty Acid Handling in Human Adipose Tissue Microvascular Endothelial Cells and Favors Inflammation. ArteriosclerThrombosisVase Biol, 2018. 38(5):pp.1134-1146).

[0012] These mechanisms of active transport of fatty acids by endothelial cells may therefore be targets for slowing down fat deposition and the entry of fatty acids into adipocytes, thereby limiting their storage in the adipose mass.

[0013] This led to the development of a cellular model of immortalized human microvascular endothelial cells from subcutaneous abdominal adipose tissue, through which the applicant demonstrated the antihyperplastic effects of an extract of Pseudoalteromonas paragorgicola. Indeed, the extract of Pseudoalteromonas paragorgicola restricted the transport of long-chain fluorescent C16 fatty acids (Bodipy FL C16), limiting the storage of FAs in adipocytes and preventing the growth of subcutaneous adipose tissue.

[0014] As a result of its properties, extracts of Pseudoalteromonas paragorgicola appear to be useful active ingredients as slimming agents. Summary of the Invention

[0015] The present invention therefore relates to the cosmetic use of an extract of Pseudoalteromonas paragorgicola as a slimming agent, more particularly as an anti-hyperplastic agent.

[0016] Cosmetic use refers to the use of a cosmetic composition, i.e. a formulation suitable for dermal application to healthy skin, comprising an extract of Pseudoalteromonas paragorgicola and cosmetically acceptable excipients.

[0017] Pseudoalteromonas paragorgicola extract can also be advantageously used in slimming compositions.

[0018] The use according to the present invention is particularly suitable for topical application to healthy skin.In the context of the present invention, healthy skin is defined as skin that does not have skin lesions and is also in good condition (within the meaning of Article 2.1.a) of Regulation (EC) No. 1223 / 2009 of the European Parliament and of the Council of November 30, 2009 on Cosmetics).Therefore, cosmetic use is understood to mean non-therapeutic use.

[0019] The use according to the invention is particularly suitable for topical application to the skin of the face, as well as for application to the skin of the body.

[0020] Pseudoalteromonas paragorgicola is a bacterium that belongs to the order Alteromonadales, more specifically the family Pseudoalteromonadiaceae.

[0021] This bacterium is a pale orange, Gram-negative, rod-shaped bacterium. It grows under strict aerobic conditions. Its diameter is 0.7-0.9 μm, length is 1-1.2 μm, and it has a single polar flagellum. Its DNA contains 41% guanine and cytosine. Circular colonies are 2-3 mm in diameter and slightly orange in color. This bacterium has the ability to degrade gelatin, DNA, and polysorbate 80. This bacterium can grow in 1-6% salinity (NaCl) or seawater. The growth temperature range is 2-30°C, with an optimum temperature of 25°C. This bacterium can grow at pH levels between 6-10, with an optimum of pH 7.5-8.

[0022] The bacterium was first isolated in 1988 from coral samples taken from a depth of 202 m in water with a salinity of 32% at 10 °C after an expedition to the Pacific Ocean, more specifically to the coastal areas of Onekotan Island (Kuril Islands, Russia). It was also collected during an expedition to international waters near Greenland (MEDEA II, June-July 2012). The sample used for isolation was taken at a depth of 2,700 m in water with a temperature of 2.98 °C.

[0023] Certain antibiotics (ampicillin, carbenicillin, polymyxin, streptomycin) are effective against this species.

[0024] The name given to this species is composed of "Paragorgia," a genus in the family Gorgonaceae, and the Latin suffix "-cola," meaning "where it lives." The species is isolated on the coral, Paragorgia arborea.

[0025] To date, there are no known conventional uses for this bacterium.

[0026] According to a particular aspect of the invention, the extract is obtained according to a method comprising at least the following steps, the skilled person knowing how to carry out an extraction method from a microorganism: fermentation Preferably hot water extraction Centrifugation Continuous filtration dialysis filtration Stabilized by the addition of 20% propanediol + 0.6% xanthan gum + 0.5% glyceryl caprylate.

[0027] The resulting Pseudoalteromonas paragorgicola extract is a gray to yellow liquid (opaque gel-like) with a characteristic odor. It has the following analytical properties: pH=4~6 Density at 20°C = 1.00-1.04 (g / ml) Dry matter=0.25~0.75% Refractive index = 1.34 to 1.38 Molecular weight=5~250kDa

[0028] The cosmetic use according to the invention preferably comprises an extract of Pseudoalteromonas paragorgicola or a cosmetic composition comprising such an extract.

[0029] The extract according to the present invention is an extract of the microorganism Pseudoalteromonas paragorgicola, more particularly, said extract is a water-soluble extract.

[0030] Advantageously, the extract of the microorganism Pseudoalteromonas paragorgicola is a water-soluble extract obtained by fermentation followed by hot aqueous extraction, purification and filtration, and stabilized with propanediol.

[0031] Preferably, the cosmetic composition according to the present invention comprises 0.01 to 10% by weight of an extract of Pseudoalteromonas paragorgicola, based on the total weight of the composition. Advantageously, the composition comprises 0.01 to 5% by weight of an extract of Pseudoalteromonas paragorgicola, based on the total weight of the composition. The composition according to the present invention may comprise one or more ingredients or additives known and conventionally used in cosmetic compositions, such as, but not limited to, emollients, colorants, film-forming active ingredients, surfactants, fragrances, preservatives, emulsifiers, oils, glycols, vitamins such as vitamin E, and UV filters. Thanks to their knowledge of cosmetics, those skilled in the art will know which ingredients to add to the composition of the present invention and in what amounts, depending on the desired properties.

[0032] The compositions according to the invention can be provided in any form known to those skilled in the art of cosmetology, without any galenical limitations other than application to the face and body, and are advantageously in the form of gels, creams, lotions, masks, oils, milks, sprays, etc.

[0033] The present invention also relates to the use of an extract of Pseudoalteromonas paragorgicola for toning and / or firming the skin, reducing hollows, improving and / or redefining the silhouette, reducing subcutaneous fat deposits, and preventing and / or delaying skin symptoms of increased adipose tissue such as cellulite and / or hollows.

[0034] The following example concerns the effect of the active ingredient Pseudoalteromonas paragorgicola on the transport of fatty acids (long chain C16) in a cell model (immortalized human abdominal subcutaneous adipose tissue microvascular endothelial cells). Example of a formulation using the Pseudoalteromonas paragorgicola extract that is the subject of the present invention. [Brief explanation of the drawings]

[0035] [Figure 1] The Examples refer to Figure 1, which shows the effect of the aforementioned Pseudoalteromonas paragorgicola extract on the transport of fluorescent C16 fatty acids. The figure represents the mean + / - standard deviation from the mean of eight duplicate determinations of cells treated with a 0.01% dose of Pseudoalteromonas paragorgicola extract as a percentage of untreated cells (control). DETAILED DESCRIPTION OF THE INVENTION [Example]

[0036] I. Demonstration of the role of Pseudoalteromonas paragorgicola in fatty acid (FA) transport A. Materials and Methods 1.Cell culture Immortalized human adipose tissue microvascular endothelial cells (CD45- / CD34+ / CD31+) were seeded to confluence (150,000 cells / cm2) on fibronectin-coated plates (2 μg / cm2, 30 min, room temperature) and allowed to adhere. Cells were maintained in growth medium (ECGM MV, i.e., endothelial cell growth medium MV; PromoCell C-22020) at 37°C in a 5% CO2 atmosphere. Cells were then rinsed with PBS and treated twice with or without 0.01% (the non-cytotoxic concentration tested) extract of Pseudoalteromonas paragorgicola for 24 h in ECGM MV growth medium.

[0037] 2. Transport of Fluorescent Fatty Acids Untreated (control) cells or cells treated with 0.01% Pseudoalteromonas paragorgicola extract were incubated for 1 hour in minimal culture medium consisting of fatty acid-free Hank's balanced salt solution (HBSS) containing 0.1% bovine serum albumin (BSA). The medium was then removed and replaced with minimal medium supplemented with fluorescent FA (BODIPY-FL C16) (Thermofisher Ref D3821; 750 nM). The cells were incubated for 1 hour. After washing with phosphate-buffered saline (PBS) preheated to 37°C, the cells were fixed with a solution of 4% paraformaldehyde (PFA) in PBS for 10 minutes at room temperature and protected from light. Fluorescence intensity was measured at 520 nm using a TECAN Infinite F500 fluorometer. Eight fields per well were measured in duplicate. Statistical analysis was performed using the paired Student's test.

[0038] 3.Toxicity To assess cytotoxicity, cells were fixed in phosphate-buffered saline (PBS) containing 4% paraformaldehyde for 10 minutes at room temperature in the dark and then rinsed with PBS. Cells were labeled with DAPI (4',6-diamidino-2-phenylindole) (20 μg / mL (Invitrogen D1306)) for 30 minutes and then rinsed with PBS at 4°C protected from light. DAPI staining allows for counting of live cells by fluorescent detection of nuclei, which are then quantified by imaging. Fluorescence intensity was measured using a TECAN Infinite F500 fluorometer, reading at 448 nm. Eight fields per well were measured in duplicate.

[0039] B. Results and Conclusions Under the selected experimental conditions, 0.01% extract of Pseudoalteromonas paragorgicola did not induce toxicity on microvascular endothelial cells from human adipose tissue treated for 24 hours.

[0040] Under selected experimental conditions, pretreatment of microvascular endothelial cells from human adipose tissue with 0.01% extract of Pseudoalteromonas paragorgicola inhibited C16 fatty acid transport. The uptake of fluorescent C16 fatty acids (BODIPY-FL C16) by cells was significantly reduced in the untreated (control) condition treated with Pseudoalteromonas paragorgicola extract (Figure 1). Pretreatment of microvascular endothelial cells from human adipose tissue with 0.01% extract of Pseudoalteromonas paragorgicola significantly inhibited C16 fatty acid transport by 10.5% ( ** , p-value=0.0047).

[0041] II. Examples of combination drugs Emulsion................................................................(%) Isononyl isononanoate................................................3.50 Dicaprylyl Carbonate / Tocopherol............................................1.50 C14-22 Alcohol / C12-20 Alkyl Glucoside ................3.00 Cetyl ethylhexanoate................................................3.50 Sodium Acrylates Copolymer / Phospholipids / Hydrogenated Polydecene / Polyglyceryl-10 Stearate / Helianthus Annus (Sunflower) Seed Oil / Tocopherol.................................................................2.50 Water...................................................65.30 Caffeine................................................................1.80 Butylene Glycol.................................................................1.50 Sodium polyacrylate.................................................0.20 Glycerin................................................................4.00 Potassium cetyl phosphate.................................................0.50 ESCIN...................................................0.20 Water / Methylsilanol Mannuronate / Sorbic Acid / Phenoxyethanol....2.00 Water / Propanediol / Xanthan Gum / Glyceryl Caprylate / Pseudoalteromonas Ferment Extract....................0.10 Fragrance...................................................0.40 Alcohol.................................................................10.00 gel....................................................................(%) Water...................................................88.37 Caffeine.................................................................0.50 Glycerin................................................................4.00 Ethylhexylglycerin.............................................0.20 Propanediol................................................1.50 Pentylene glycol.................................................1.00 Ammonium acryloyldimethyltaurate / VP copolymer.............0.70 Polyacrylate crosspolymer-6 / water / t-butyl alcohol..........0.35 ESCIN...................................................0.10 Water...................................................0.08 Sodium hydroxide................................................0.10 Water / Oat (AVENA SATIVA) Kernel Extract / Sodium Benzoate 2.00 Water / Propanediol / Xanthan Gum / Glyceryl Caprylate / Pseudoalteromonas Ferment Extract....................0.10 Glycerin / Water / Panax Ginseng Root Extract 1.00

Claims

1. Cosmetic use of an extract of Pseudoalteromonas paragorgicola as a slimming agent.

2. Cosmetic use of an extract of Pseudoalteromonas paragorgicola as a slimming and anti-hyperplastic agent.

3. Cosmetic use of an extract of Pseudoalteromonas paragorgicola in a slimming composition.

4. 4. The use according to any one of claims 1 to 3, wherein the extract of Pseudoalteromonas paragorgicola is a water-soluble extract obtained by fermentation.

5. 5. The use according to any one of claims 1 to 4, wherein the extract of Pseudoalteromonas paragorgicola is stabilized by propanediol.

6. 6. Use according to any one of claims 1 to 5, characterized in that the extract is formulated in a cosmetic composition comprising from 0.01 to 10% by weight of an extract of Pseudoalteromonas paragorgicola relative to the total weight of the composition, preferably from 0.01 to 5% by weight of said extract relative to the total weight of the composition.

7. 7. The use according to claim 3 or 6, wherein the composition further comprises one or more ingredients or additives known and conventionally used in cosmetic compositions, such as, by way of example and not limitation, emollients, colorants, film-forming active ingredients, surfactants, fragrances, preservatives, emulsifiers, oils, glycols, vitamins such as vitamin E, and filters.

8. 8. Use according to any one of claims 1 to 7 for toning and / or firming the skin, reducing hollows, improving and / or redefining its shape, reducing subcutaneous fat deposits, preventing and / or delaying skin symptoms of increased adipose tissue such as cellulite and / or hollows.

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