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

Abstract

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

Description

Cosmetic Use of a Pseudoalteromonas paragorgicola Extract as a Slimming Agent. The present invention relates to the field of cosmetic skin treatment. It relates to a cosmetic composition comprising a Pseudoalteromonas paragorgicola extract. The skin is a superficial covering organ that forms a relatively effective protective barrier against external aggressions and covers the entire body surface. It is often considered the heaviest organ in the human body. Furthermore, the skin also plays an active role in numerous biological and biochemical processes. It is made up of three superimposed layers or tissues that comprise, from the most superficial to the deepest part, the epidermis, the dermis and the hypodermis. This tissue, also called adipose tissue, represents between 15 and 20% of body weight in an average-weight person. It is attached to the lower part of the dermis by extensions of collagen fibers. The thickness of the hypodermis varies according to age, body area, and sex, and largely determines body contouring. In addition to being a vascularized adipose tissue containing adipocytes, whose function is to absorb fats and store them as triglycerides, it is a storage organ where fats can be mobilized and transformed into energy during prolonged exertion. It also acts as a protective organ, providing mechanical protection against impacts and thermal protection against temperature variations to maintain body homeostasis. It controls lipid mobilization through two main metabolic activities: lipogenesis (synthesis of fatty acids (FAs) and triglycerides, and storage of excess energy) and lipolysis (hydrolysis of triglycerides when energy is needed). Adipose tissue is not merely a storage organ whose excessive development leads to obesity and associated metabolic disorders, such as type 2 diabetes; it is also a true endocrine organ. This function was attributed to it more than 20 years ago with the discovery of leptin, a hormone secreted by adipose tissue in response to food intake that inhibits appetite by regulating neural circuits located in the brain. Adipose tissue is a fundamental organ for maintaining the body's energy homeostasis thanks to its capacity to store and release fatty acids, as well as its secretory functions. The endothelial cells of the adipose tissue's vascular network form a physical barrier, called the endothelium, between the bloodstream and the adipocytes. These endothelial cells play a crucial role in controlling the exchange of nutrients, hormones, oxygen, and immune and inflammatory cells. These exchanges and their regulation are essential and specific to the metabolic state of the adipose tissue. The role of the endothelium in the biology of adipose tissue has evolved considerably since its historical conception as an inert layer of cells serving as an internal lining for the poorly developed circulatory system within the fat mass. Current data show, on the one hand, that the vascular network is very extensive in adipose tissue and, on the other hand, that it plays a key role in the development and function of adipose tissue. Furthermore, the remarkable heterogeneity of the endothelial layer is now recognized, whether due to differences based on the location of the conducting vessels (arteries, capillaries, and veins) or to functional heterogeneity in the context of specific organs, such as the heart, liver, and adipose tissue. This diversity of the endothelium is key to its specialized functions, which include permeability, leukocyte trafficking, and hemostasis, as well as the transport of oxygen, fluids, and nutrients from the circulation to target tissues. The concept of fatty acid (FA) transport by the endothelium has existed for many years, but it has evolved from a passive, diffuse transfer to an active, highly regulated, and tissue-specific process involving multiple complex signaling pathways. (Hagberg, CE, et al., Vascular endothelial growth factor B controls endothelial fatty acid uptake. Nature, 2010. 464 (7290): pp. 917-21). Fatty acids (FAs), naturally stored as triglycerides in adipocytes, must be transported across the microvascular endothelium via mechanisms that are not yet fully understood.The transport of fatty acids (FAs) across the non-fenestrated microvascular endothelium involves membrane transporters FATP (fatty-acid-transport proteins) and cytoplasmic transporters FABP (fatty-acid-binding proteins). In the endothelium of white adipose tissue (WAT), other factors, including PPARG (peroxisome proliferator-activated receptor gamma), have been proposed to play an important role. To analyze GA transport in human adipose tissue endothelial cells (hATECs), Anaïs Briot's team measured the ability of freshly isolated primary endothelial cells from adipose tissue to absorb a fluorescent GA (Bodipy FL C16) in vitro. At 37 °C, the cells efficiently incorporate the fluorescent GAs. In the presence of rosiglitazone, a PPARγ agonist, at 37 °C, an increase in the absorption of fluorescent GAs was observed, demonstrating that rosiglitazone treatment favors GA absorption in the endothelial cells of human adipose tissue (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. Arterioscler Thromb Vasc Biol, 2018.38 (5) : pp.1134-1146). Therefore, these mechanisms of active transport of fatty acids by endothelial cells may be a target to slow the entry of fatty acids into fat deposits and into adipocytes, and thus limit their storage in fat masses. This enabled the development of a cell model of immortalized human abdominal subcutaneous adipose microvascular endothelial cells, through which the applicant demonstrated the antihyperplastic effect of a Pseudoalteromonas paragorgicola extract. Specifically, the Pseudoalteromonas paragorgicola extract limits the transport of C16 long-chain fluorescent fatty acids (C16 FL), thereby restricting fatty acid storage in adipocytes and exerting a preventative effect on subcutaneous adipose tissue accumulation. Due to its properties, the extract of Pseudoalteromonas paragorgicola appears to be a useful resource as a slimming agent. Several prior art documents propose cosmetic slimming agents: Patent FR2849595 describes the use of the plant Bocoa prouacensis as a slimming agent; Patent FR2921267 proposes using a perilipin activator as a plant extract from the Dipsacaceae family in a slimming cosmetic composition; and Patent WO2005 / 087189 describes the use of a metalloproteinase 2 and / or 9 inhibitor, such as an extract of Baccharis genistelloides, in a slimming cosmetic composition. None of these documents suggest the slimming effect of an extract of Pseudoalteromonas paragorgicola. Therefore, the present invention relates to the cosmetic use of an extract of Pseudoalteromonas paragorgicola as a slimming agent. Cosmetic use is understood to mean the use of a cosmetic composition, that is, a formulation suitable for application on healthy skin, comprising the extract of Pseudoalteromonas paragorgicola as well as cosmetically acceptable excipients. The extract of Pseudoalteromonas paragorgicola can also be used advantageously in a slimming composition. The use according to the present invention is suitable, in particular, for topical application to healthy skin. For the purposes of this invention, healthy skin is defined as skin that is free from skin pathologies and 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 30 November 2009 on cosmetic products). Therefore, cosmetic use is understood to mean non-therapeutic use. The use according to the present invention is suitable, in particular, for topical application to the skin of the face, as well as for application to the skin of the body. Pseudoalteromonas paragorgicola is a bacterium that belongs to the order Alteromonadales, and more specifically to the family Pseudolateromonadaceae. This is a pale orange, Gram-negative, rod-shaped bacterium. It grows under strict aerobic conditions. It measures between 0.7 and 0.9 µm in diameter and between 1 and 1.2 µm in length, with a single polar flagellum. Its DNA consists of 41% guanine and cytosine. The colony is round, measures 2 to 3 mm in diameter, and is slightly orange. This bacterium can degrade gelatin, DNA, and polysorbate 80. It can grow in salt (NaCl) concentrations of 1 to 6% or in seawater. Its growth range is between 2 and 30 °C, with an optimum temperature of 25 °C. This bacterium can grow at a pH between 6 and 10, with an optimum pH of 7.5–8. It was first isolated in 1988 from a coral sample collected at a depth of 202 m, in water at 10 °C and a salinity of 32, following an expedition in the Pacific Ocean, specifically off the coast of Onekotan Island (Kuril Islands archipelago, Russia). It was also collected during an expedition in international waters near Greenland (MEDEA II, June–July 2012). The sample that allowed its isolation was taken at a depth of 2,700 m, in water at 2.98 °C. Some antibiotics (ampicillin, carbenicillin, polymyxin, streptomycin) are effective against this species. The species name is derived from "Paragorgia," a genus in the Gorgonaceae family, and the Latin suffix "-cola," meaning "inhabitant." In fact, the species was isolated from the coral Paragorgia arborea. To date, no traditional uses for this bacterium have been recorded. According to a particular embodiment of the present invention, the extract is obtained by a process comprising at least the following steps; a person skilled in the art knows how to carry out an extraction process from the microorganism: - Fermentation - Extraction, preferably with hot water - Centrifugation - Successive leaks - Dialysis - Filtration - Stabilization by the addition of 20% propanediol + 0.6% xanthan gum + 0.5% glyceryl caprylate. The Pseudoalteromonas paragorgicola extract obtained in this way is a gray to yellow liquid (opaque gel type) with a characteristic odor. It has the following analytical characteristics: - pH = 4 - 6 - density at 20 °C = 1.0.- 1.04 (g / ml) - dry matter = 0.2 - 0.75% - refractive index = 1.3 - 1.38 - molecular weight = 5-250 kDa Cosmetic use according to the present invention preferably involves an extract of Pseudoalteromonas paragorgicola or a cosmetic composition comprising such an extract. The extract according to the present invention is an extract of the microorganism Pseudoalteromonas paragorgicola, more specifically, said extract is a water-soluble extract. Advantageously, the extract of the microorganism Pseudoalteromonas paragorgicola is a water-soluble extract obtained by fermentation, followed by hot aqueous extraction, purifying filtrations, and is stabilized with propanediol. Preferably, the cosmetic composition according to the present invention comprises from 0.01 to 10% of a Pseudoalteromonas paragorgicola extract by weight of the total composition. Advantageously, the composition comprises from 0.01 to 5% of a Pseudoalteromonas paragorgicola extract by weight of the total composition. The compositions according to the present invention may comprise one or more formulation agents or additives of known and conventional use in cosmetic compositions, such as, by way of example and without limitation, emollients, colorants, film-forming agents, surfactants, perfumes, preservatives, emulsifiers, oils, glycols, vitamins such as vitamin E, UV filters, etc. Thanks to their knowledge of cosmetics, a person skilled in the art will know which formulation agents to add to the compositions of the present invention and in what quantities, depending on the desired properties. The compositions according to the present invention can be presented in any form known to those skilled in the art of cosmetology, with no other pharmaceutical restriction than their application to the face and body. Advantageously, the compositions according to the present invention are presented in the form of a gel, cream, lotion, mask, oil, milk, spray, etc. The present invention also relates to the use of an extract of Pseudoalteromonas paragorgicola to tone and / or firm the skin, reduce orange peel skin, refine and / or redefine the silhouette, reduce subcutaneous fat deposits, prevent and / or delay the skin manifestations of adipose tissue accumulation, such as cellulite and / or orange peel skin. The following examples refer, on the one hand, to the impact of the active agent Pseudoalteromonas paragorgicola on the transport of fatty acids (long chain C16) in a cell model (immortalized human abdominal subcutaneous adipose microvascular endothelial cells), and on the other hand, to examples of formulations using the Pseudoalteromonas paragorgicola extract that is the subject of the present invention. Figure 1: The examples refer to Figure 1, which shows the effect of a Pseudoalteromonas paragorgicola extract, as described above, on the transport of fluorescent C16 fatty acids. This figure represents the mean ± standard deviation of the mean of 8 duplicate measurements of cells treated with the Pseudoalteromonas paragorgicola extract at the reported dose of 0.01% relative to untreated (control) cells. EXAMPLES I. Demonstration of the role of Pseudoalteromonas paragorgicola on fatty acid (FA) transport A. Materials and Methods 1. Cell culture Immortalized human adipose tissue microvascular endothelial cells (CD45- / CD34+ / CD31+) are seeded to confluence (150,000 cells / cm²) in plates coated with fibronectin (2 µg / cm², 30 minutes, room temperature) to promote cell adhesion. The cells are maintained in culture medium (ECGM MV or endothelial cell growth medium MV; PromoCell C-22020) at 37 °C under a 5% CO₂ atmosphere. The cells are then rinsed with PBS and subsequently treated, or not, in duplicate with 0.01% Pseudoalteromonas paragorgicola extract (non-cytotoxic concentration tested) in ECGM MV for 24 h. 2. Transport of fluorescent fatty acids Untreated cells (control) or cells treated with Pseudoalteromonas paragorgicola extract at a dose of 0.01% are incubated in a minimal culture medium consisting of Hank's Balanced Salt Solution (HBSS) with 0.1% fatty acid-free bovine serum albumin (BSA) for 1 hour. The medium is then discarded and replaced with a minimal medium supplemented with a fluorescent AG (BODIPY-FL C16) (Thermofisher, ref. D3821; 750 nM). The cells are incubated for 1 hour. After washing with phosphate-buffered saline (PBS) preheated to 37 °C, the cells are fixed with PBS containing 4% paraformaldehyde (PFA) for 10 minutes at room temperature and protected from light. Fluorescence intensity is measured using a TECAN Infinite F500 fluorometer, with a reading at a wavelength of 520 nm. Eight fields per well are measured in duplicate.Statistical analysis is performed using a paired Student's t-test. 3. Toxicity To assess cell toxicity, cells are fixed in phosphate-buffered saline (PBS) containing 4% paraformaldehyde for 10 minutes at room temperature, protected from light, and then rinsed with PBS. Cells are labeled with DAPI or 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 enumeration of viable cells by detecting nuclei using fluorescence, which are then quantified by imaging. Fluorescence intensity is measured using a TECAN Infinite F500 fluorometer, with a reading at 448 nm. Eight fields per well are measured in duplicate. B. Results and Conclusion Under the experimental conditions maintained, the 0.01% Pseudoalteromonas paragorgicola extract did not induce toxicity in microvascular endothelial cells of human adipose deposits treated for 24 hours. Under the experimental conditions maintained, pretreatment of microvascular endothelial cells from human adipose tissue with 0.01% Pseudoalteromonas paragorgicola extract inhibits C16 fatty acid transport. Specifically, the uptake of the fluorescent C16 fatty acid (BODIPY-FL C16) by the cells in the Pseudoalteromonas paragorgicola extract-treated condition is significantly reduced compared to the untreated (control) condition (Figure 1). Pretreatment of microvascular endothelial cells from human adipose tissue with 0.01% Pseudoalteromonas paragorgicola extract significantly inhibits (**, p-value = 0.0047) C16 fatty acid transport by 10.5%. II. Example of formulation EMULSION (%) ISONONYL ISONONANOATE 3.50 DICAPRYL CARBONATE / TOCOPHEROL 1.50 C14-22 ALCOHOLS / C12-20 ALKYL GLUCOSIDE 3.00 CETYL ETHYLHEXANOATE 3.50 SODIUM ACRYLATES COPOLYMER / PHOSPHOLIPIDS / HYDROGENATED POLYDECENE / 2.50 POLYGLYCERYL-10 STEARATE / HELIANTHUS ANNUUS (SUNFLOWER) SEED OIL / TOCOPHEROL WATER 65.30 CAFFEINE 1.80 BUTYLENE GLYCOL 1.50 SODIUM POLYACRYLATE 0.20 GLYCERIN 4.00 CETYLPOTASIUM PHOSPHATE 0.50 ESCINE 0.20 WATER / METHYLSILANOL MANURONATE / SORBIC ACID / PHENOXYETHANOL 2.00 WATER / PROPANEDIOL / XANTHAN GUM / GLYCERYL CAPRYLATE / PSEUDOALTEROMONAS FERMENT EXTRACT 0.10 PERFUME 0.40 ALCOHOL 10.00 GEL (%) WATER 88.37 CAFFEINE 0.50 GLYCERIN 4.00 ETHYLHEXYLGLYCERIN 0.20 PROPANADIOL 1.50 PENTYLENE GLYCOL 1.00 AMMONIUM ACRYLOYLDIMETHYLTAURATE COPOLYMER / VP 0.70 POLYACRYLATE-6 CROSSPOLYMER / WATER / t-BUTYL ALCOHOL 0.35 ESCINE 0.10 WATER 0.08 SODIUM HYDROXIDE 0.10 WATER / OAT (Avena SATIVA) KERNEL EXTRACT / SODIUM BENZOATE 2.00 WATER / PROPANADIOL / XANTHAN GUM / GLYCERYL CAPRYLATE / FERMENT EXTRACT 0.10 PSEUDOALTEROMONAS 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 in a slimming composition.

3. Use according to any of the preceding claims, wherein the extract of Pseudoalteromonas paragorgicola is a water-soluble extract obtained by fermentation.

4. Use according to any of the preceding claims, wherein the extract of Pseudoalteromonas paragorgicola is stabilized with propanediol.

5. Use according to any of the preceding claims, characterized in that said extract is formulated in a cosmetic composition comprising from 0.01 to 10% by weight of the 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. 6.Use according to claim 2 or 5, wherein said composition further comprises one or more formulation agents or additives of known and conventional use in cosmetic compositions such as, by way of example and not limitation, emollients, colorants, film-forming agents, surfactants, perfumes, preservatives, emulsifiers, oils, glycols, vitamins such as vitamin E, and filters.

7. Use according to any of the preceding claims for toning and / or firming the skin, reducing the appearance of cellulite, refining and / or redefining the silhouette, reducing subcutaneous fat deposits, and preventing and / or delaying the cutaneous manifestations of adipose tissue accumulation, such as cellulite and / or orange peel skin.