Cosmetic composition comprising pseudoalteromonas corallina extract

Abstract

The present invention relates to the cosmetic use of an extract of Pseudoalteromonas corallina as a weight-reducing agent, and more particularly to the use of an extract of Pseudoalteromonas corallina as an anti-proliferative agent. The invention also relates to an application of the extract of the pseudoalteromonas corallina in the weight-losing composition.

Description

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

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

[0003] The skin is composed of three overlapping layers or tissues which, from the most superficial to the deepest part, include the epidermis, dermis, and subcutaneous tissue.

[0004] This subcutaneous tissue, also called adipose tissue, accounts for 15-20% of the body weight in an average individual. It is attached to the lower part of the dermis by collagen fiber expansion. The thickness of the subcutaneous tissue varies according to age, body position and gender, and largely determines the shaping of the body curves.

[0005] In addition to being a vascularized adipose tissue containing adipocytes whose role is to absorb fat and store it in the form of triglycerides, the subcutaneous tissue is also a reserve organ that enables fat to be mobilized and converted into energy in the event of prolonged exercise of the organism. It also acts as a protective organ that mechanically resists shocks and thermally resists temperature changes to maintain the body's homeostasis. It controls the mobilization of lipids through two main metabolic activities: lipogenesis (synthesis of fatty acids (FA) 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, the overdevelopment of which is the cause of obesity and its accompanying metabolic disorders (such as type II diabetes); it is also a true endocrine organ. The discovery of leptin more than 20 years ago gave it the following function: leptin is a hormone secreted by adipose tissue in response to food intake, which suppresses appetite by regulating neural circuits in the brain.

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

[0008] The role of endothelial cells in the biology of fat deposits has evolved considerably from its historical role as an inert cell layer that coats the inner layer of the poorly developed circulatory system in the fat mass. Current data indicate that the vascular network is extensive in fat deposits and plays a key role in the development and function of adipose tissue. Furthermore, the remarkable heterogeneity of the endothelium is now well recognized, from differences in the location of conducting blood vessels (arteries, capillaries, and veins) to the functional heterogeneity of specific organs such as the heart, liver, and adipose tissue.

[0009] This diversity of endothelial cells is key to their specialized roles, including permeability, leukocyte trafficking and hemostasis, and the transport of oxygen, fluids and nutrients from the circulation to target tissues. The concept of fatty acid (FA) transport through endothelial cells has been around for many years, but it has evolved from a passive, diffusive transport to an active, highly regulated, 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): p.917-21).

[0010] FA is naturally stored in adipocytes as triglycerides and must be directed through microvascular endothelial cells according to mechanisms that are currently unclear. Transport of FA through fenestrated microvascular endothelial cells involves the membrane transporter FATP (fatty-acid-transport protein) and the cytoplasmic transporter FABP (fatty-acid-binding protein). In endothelial cells of white adipose tissue (WAT), other factors including peroxisome proliferator-activated receptor gamma (PPARG) are thought to play a role.

[0011] To analyze FA transport in endothelial cells of the human adipose tissue (hATEC), Briot's team measured the ability of freshly isolated primary adipose tissue endothelial cells to absorb fluorescent FA (Bodipy FL C16) in vitro. At 37 ° C, cells effectively bind fluorescent FA. In the presence of the PPARG agonist rosiglitazone at 37 ° C, an increase in fluorescent FA absorption was observed, indicating that treatment with rosiglitazone promoted FA absorption in human adipose tissue endothelial cells (Briot, A. Senescence Alters PPARgamma (Peroxisome Proliferator-Activated ReceptorGamma)-Dependent Fatty Acid Handling in Human Adipose Tissue Microvascular Endothelial Cells and Favors Inflammation. ArteriosclerThromb Vase Biol, 2018. 38 (5): p. 1134-1146).

[0012] Therefore, these active transport mechanisms of fatty acids by endothelial cells could be targeted to slow the influx of fatty acids into fat deposits and adipocytes, thereby limiting their storage in the adipose mass.

[0013] This has made it possible to develop a cell model of immortalized microvascular endothelial cells in the subcutaneous adipose tissue of the human abdomen, through which the Applicant demonstrated the antiproliferative effect of an extract of Pseudoalteromonas corallidense. In fact, the extract of Pseudoalteromonas corallidense limits the transport of long-chain fluorescent C16 fatty acids (Bodipy FL C16), making it possible to limit the storage of FA in adipocytes and prevent the increase of subcutaneous adipose tissue.

[0014] Due to its properties, extracts of Pseudoalteromonas coralligensis appear to be useful active ingredients of slimming agents.

[0015] Thus, the present invention relates to the cosmetic use of an extract of Pseudoalteromonas corallidans as a slimming agent, and more specifically, as an antihyperplasia agent.

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

[0017] Extracts of Pseudoalteromonas corallids can also be advantageously used in weight loss compositions.

[0018] The use according to the 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 show skin pathology 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 30 November 2009 on cosmetic products). 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 facial skin and for application to the body skin.

[0020] Pseudoalteromonas corallidense is a bacterium belonging to the order Alteromonadales, and more specifically to the family Pseudolateromonadaceae.

[0021] It is a light orange Gram-negative "rod-shaped" bacterium. It develops under strict aerobic conditions. Its diameter measures 0.7 to 0.9 μm and its length is 1 to 1.2 μm, with a unique polar flagellum. Its DNA consists of 41% guanine and cytosine. The circular colonies measure 2 to 3 mm in diameter and are slightly orange. The bacterium has the ability to degrade gelatin, DNA and polysorbate 80. It can grow at a salt concentration (NaCl) of 1 to 6% or in seawater. The temperature range in which the bacterium can proliferate is 2 to 30°C, with an optimum temperature of 25°C. The bacterium can grow at pH levels between 6 and 10, with an optimum pH of 7.5-8.

[0022] The bacterium was first isolated in 1988 from a coral sample at a depth of 202 m, in water of 10 ° C and a salinity of 32% after an expedition in the Pacific Ocean, specifically off the coast of Onekotan island (Russian Kuril Islands). It was also collected during an expedition to international waters near Greenland (MEDEA II, June-July 2012). The sample used to isolate it was collected at a depth of 2,700 m and in water of 2.98 ° C.

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

[0024] The species' name is a combination of "Paragorgia", a genus in the Gorgonaceae family, and the Latin suffix "-cola", meaning "dwelling". The species has been isolated from the coral Pararagorgia arborea.

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

[0026] According to a particular mode of the invention, the extract is obtained according to a method comprising at least the following steps, how to achieve extraction from microorganisms is known to those skilled in the art:

[0027] Fermentation

[0028] Optimum hot water extraction

[0029] Centrifugation

[0030] Continuous filtration

[0031] Dialysis

[0032] filter

[0033] Stabilized by addition of 20% propylene glycol + 0.6% xanthan gum + 0.5% glyceryl caprylate.

[0034] The obtained extract of Pseudoalteromonas corallidense is a gray to yellow liquid (opaque gelatinous) with a distinctive odor. It has the following analytical characteristics:

[0035] pH = 4–6

[0036] Density at 20℃=1.00-1.04(g / ml)

[0037] Dry matter = 0.25-0.75%

[0038] Refractive index = 1.34-1.38

[0039] Molecular weight = 5-250 kDa

[0040] The cosmetic use according to the invention preferably relates to an extract of Pseudoalteromonas corallidans or a cosmetic composition comprising such an extract.

[0041] The extract according to the present invention is an extract of the microorganism Pseudoalteromonas corallii, more specifically, the extract is a water-soluble extract.

[0042] Advantageously, the extract of the microorganism Pseudoalteromonas corallidans is a water-soluble extract obtained by fermentation followed by hot water extraction, purification filtration and stabilization with propylene glycol.

[0043] Preferably, the cosmetic composition according to the invention comprises an extract of Pseudoalteromonas corallidans in an amount of 0.01% to 10% by weight relative to the total weight of the composition. Advantageously, the composition comprises an extract of Pseudoalteromonas corallidans in an amount of 0.01% to 5% by weight relative to the total weight of the composition. The composition according to the invention may comprise one or more formulation agents or additives known and conventionally used in cosmetic compositions, such as, by way of example and not limitation, softeners, colorants, film-forming active ingredients, surfactants, perfumes, preservatives, emulsifiers, oils, glycols, vitamins (such as vitamin E), UV filters, etc. Thanks to the knowledge of the skilled person in the field of cosmetics, they will know which formulation agents to add to the composition of the invention and in what amounts, depending on the desired properties.

[0044] The composition according to the invention may be presented in any form known to those skilled in the cosmetic field, without galenic restriction other than application to the face and body. Advantageously, the composition according to the invention is in the form of a gel, cream, lotion, mask, oil, milk, spray, etc.

[0045] The present invention also relates to the use of an extract of Pseudoalteromonas corallidans for toning and / or tightening the skin, reducing depressions, improving and / or redefining the body shape, reducing subcutaneous fat deposits, preventing and / or delaying the cutaneous manifestations of increased adipose tissue (such as cellulite and / or dimpling).

[0046] The following examples relate to the effect of the active ingredient Pseudoalteromonas anguillica on the transport of fatty acids (long-chain C16) in a cell model (microvascular endothelial cells of immortalized human abdominal subcutaneous adipose tissue).Examples of preparations using extracts of Pseudoalteromonas anguillica that are the subject of the present invention.

[0047] Figure 1 :These embodiments refer to Figure 1 , which shows the effect of the extract of Pseudoalteromonas coral-dwelling on fluorescent C16 fatty acid transport as described previously. The figure shows the mean + / - standard deviation of 8 replicate measurements of cells treated with a 0.01% dose of the extract of Pseudoalteromonas coral-dwelling compared to the percentage of untreated cells (control group). Example

[0048] I. Role of coral-dwelling Pseudoalteromonas in fatty acid transport

[0049] A. Materials and Methods

[0050] 1. Cell Culture

[0051] Immortalized human adipose tissue microvascular endothelial cells (CD45- / CD34+ / CD31+) were grown at confluence (150,000 cells / cm 2 ) were inoculated on a fibronectin-coated (2 μg / cm 2 , 30 minutes, room temperature) on a plate to allow adhesion. The cells were maintained in culture medium (ECGM MV or endothelial cell growth medium MV; PromoCell C-22020) at 37°C in a 5% CO2 atmosphere. The cells were then rinsed with PBS and treated or not with 0.01% (the non-cytotoxic concentration studied) of an extract of Pseudoalteromonas corallidans in ECGM MV growth medium for 24 hours.

[0052] 2. Transport of fluorescent fatty acids

[0053] Untreated cells (control group) or cells treated with 0.01% extract of Pseudoalteromonas corallidans were incubated for 1 hour in a minimal medium consisting of Hank's Balanced Salt Solution (HBSS) and 0.1% fatty acid-free bovine serum albumin (BSA or bovine serum albumin). The medium was then removed and replaced with minimal medium supplemented with a fluorescent FA (BODIPY-FL C16) (Thermofisher reference D3821; 750 nm). The cells were incubated for 1 hour. After washing with phosphate buffered saline or PBS (phosphate buffer saline) preheated to 37°C, the cells were fixed with a PBS solution containing 4% paraformaldehyde or PFA for 10 minutes at room temperature, protected from light. The fluorescence intensity was measured with a TECAN Infinite F500 fluorimeter, reading at 520 nm. The measurements were repeated for 8 fields per well. Statistical analysis was performed using a matched Student's test.

[0054] 3. Toxicity

[0055] To evaluate cytotoxicity, cells were fixed in phosphate buffered saline or PBS containing 4% paraformaldehyde, fixed for 10 minutes at room temperature in the dark, and then rinsed with PBS. Cells were labeled with DAPI or 4',6-diamino-2-phenylindole (20 μg / mL (Invitgen D1306)) for 30 minutes, and then rinsed with PBS at 4°C in the dark. DAPI staining enables counting of living cells by fluorescence detection of the nucleus, which is then quantified by imaging. Fluorescence intensity was measured with a TECAN Infinite F500 fluorometer, read at 448 nm. The measurement was repeated for 8 fields per well.

[0056] B. Results and Conclusions

[0057] Under the chosen experimental conditions, 0.01% of the extract of Pseudoalteromonas corallidans did not induce toxicity in human adipose-deposited microvascular endothelial cells treated for 24 hours.

[0058] Under the selected experimental conditions, pretreatment of human adipose depot microvascular endothelial cells with 0.01% extract of Pseudoalteromonas coralligensis inhibited the transport of C16 fatty acids. The uptake of fluorescent C16 fatty acids (BODIPY-FL C16) by cells treated with Pseudoalteromonas coralligensis extract was significantly reduced compared to untreated (control) conditions ( Figure 1 Pretreatment of human adipose-depositing microvascular endothelial cells with 0.01% of the extract of Pseudoalteromonas corallids significantly inhibited the transport of C16 fatty acids by 10.5% (**, p value = 0.0047).

[0059] II. Examples of Formulations

[0060] Emulsion (EMULSION) (%)

[0061] 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 Annuus (Sunflower) Seed Oil / Tocopherol........................2.50 Water .................................65.30 Caffeine................................1.80 Butanediol.......................1.50 Sodium polyacrylate...............0.20 Glycerin......................4.00 Potassium cetyl phosphate..................0.50 Escin........................0.20 Water / Methylsilanol Mannosuronate / Sorbic Acid / Phenoxyethanol 2.00 Water / Propylene Glycol / Xanthan Gum / Glyceryl Caprylate / Pseudoalteromonas fermentation product extract.......................0.10 Fragrance (PARFUM)......................0.40 Alcohol......................10.00 gel...................(%) Water......................88.37 Caffeine.....................0.50 Glycerin.....................4.00 Ethylhexylglycerin........................0.20 Propylene glycol................1.50 Pentylene glycol................1.00 Ammonium acryloyldimethyltaurate / VP copolymer..............0.70 Polyacrylate crosspolymer-6 / water / tert-butyl alcohol....................0.35 Escin.......................0.10 Water......................0.08 Sodium hydroxide......................0.10 Water / Oat Kernel Extract / Sodium Benzoate..................2.00 Water / Propylene Glycol / Xanthan Gum / Glyceryl Caprylate / Pseudoalteromonas fermentation product extract.......................0.10 Glycerin / Water / Ginseng Root (PANAX GINSENG) Extract......................1.00.

Claims

1. A cosmetic use of an extract of Pseudoalteromonas corallii as a slimming agent.

2. Cosmetic use of an extract of Pseudoalteromonas corallii as an anti-obesity and anti-proliferative agent.

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

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

5. The use according to any one of the preceding claims, wherein The Pseudoalteromonas corallid extract is stabilized with propylene glycol.

6. Use according to any one of the preceding claims, characterized in that The extract is formulated in the form of a cosmetic composition comprising 0.01 to 10% by weight of the extract of Pseudoalteromonas corallidans relative to the total weight of the composition, preferably 0.01 to 5% by weight of the extract relative to the total weight of the composition.

7. The use according to claim 3 or 6, wherein The composition additionally comprises one or more formulating agents 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. Use according to any one of the preceding claims for toning and / or tightening the skin, reducing depressions, improving and / or redefining the body shape, reducing subcutaneous fat deposits, preventing and / or delaying the skin clinical manifestations of increased adipose tissue, such as cellulite and / or depressions.

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