Fermented extract of nannochloropsis and cosmetic and dermatological uses

Abstract

The present invention provides a fermented extract of Nannochloropsis Oculata offering demonstrated benefits for the skin, including anti-aging, anti-inflammatory, antioxidant, soothing, moisturizing and probiotic effects, among others. The invention also relates to the cosmetic and / or dermatological uses of the extract and to compositions incorporating said extract.

Description

[0001] Description

[0002] Title: Fermented extract of Nannochloropsia and cosmetic and dermatological uses. Technical field of the invention.

[0003] The present invention relates to the cosmetic and dermatological field, and in particular to fermented extracts of microalgae. More specifically, the invention concerns a fermented Nannochloropsis extract exhibiting excellent properties, including anti-inflammatory, anti-aging, antioxidant, soothing, repairing, and moisturizing properties, among others.

[0004] STATE OF THE ART

[0005] The rise of the cosmetics industry and the growing interest in skincare have led to the development of numerous products designed to improve skin health. A large proportion of these formulations rely on synthetic or chemical ingredients, which can sometimes present challenges in terms of incorporation and side effects such as eye irritation or itching. Consequently, the demand for cosmetics formulated with natural ingredients continues to grow, particularly to address concerns related to skin microbiome balance, sensitive skin, and dry skin.

[0006] Microalgae are a rich natural source of bioactive molecules with beneficial properties for the skin. Their diverse nutritional profile nourishes the skin and supports cellular health. Some of their bioactive compounds, particularly polyphenols, possess antioxidant properties that neutralize free radicals and protect the skin against oxidative stress and the effects of UV rays. Furthermore, microalgae's high content of hydrating compounds promotes water retention in the epidermis, thus limiting dehydration and improving skin smoothness. Certain microalgae fractions also exhibit anti-inflammatory effects, potentially helping to soothe sensitive skin. Finally, algal pigments such as 3-carotene can contribute to brightening the complexion and reducing pigmentation irregularities.Fermentation is a biotechnological approach that optimizes the effectiveness of active compounds derived from microalgae. This process enhances the bioavailability of molecules by facilitating their enzymatic degradation by microorganisms, thereby improving their skin absorption and biological effect. Furthermore, microalgae cultivation and fermentation are environmentally responsible processes, requiring little energy and generating minimal waste. Microalgae contribute to approximately 40% of global photosynthesis and play a key role in carbon sequestration, thus contributing to a sustainable and renewable production model.

[0007] Despite the rise of fermented ingredients in cosmetics, active ingredients derived from microalgae remain underrepresented on the market. To date, only a few fermented extracts, such as Chlorella ferments, are commonly used in cosmetic products, primarily for their moisturizing, anti-aging, illuminating, and antioxidant properties. Indeed, although theoretically several microalgae ferments could have beneficial effects on the skin, the results are inconsistent and the resulting products are not always effective.

[0008] DESCRIPTION OF THE INVENTION

[0009] The present invention provides a fermented extract of Nannochloropsis offering proven benefits for the skin, including anti-aging, anti-inflammatory, antioxidant, soothing, anti-eczema, and probiotic effects, among others. In particular, the invention relates to a fermented microalgae extract characterized in that it comprises a Nannochloropsis ferment, preferably Nannochloropsis oculata.

[0010] By Nannochlorpsis ferment we mean an extract containing soluble fractions from the fermentation of said microalga and / or residues of fermented biomass.

[0011] According to one embodiment, the ferment is obtained through predominantly alcoholic fermentation, lactic acid fermentation, or alcoholic and acetic acid fermentation of Nannochloropsis oculata. In a preferred embodiment, the extract is fermented by a yeast of the genus Saccharomyces via predominantly alcoholic fermentation. The extract thus obtained exhibits at least one of the following characteristics:

[0012] - a dry matter content w / w greater than 1.4%, preferably greater than 1.6%;

[0013] - a pH between 3 and 4; and

[0014] - a polyphenol content of at least 0.30 g / L, said polyphenols being calculated as gallic acid equivalents.

[0015] The invention also relates to compositions and products incorporating the extract of the invention into their composition.

[0016] The invention relates in particular to a cosmetic or dermatological composition comprising, in a cosmetically or dermatologically acceptable medium, a fermented extract according to the invention, and preferably comprising at least one adjuvant selected from the group consisting of antioxidants, emollients, moisturizing agents, anti-aging agents, preservatives, diluting and / or solubilizing agents, perfumes, and mixtures thereof. "Cosmetically or dermatologically acceptable" means what is useful in the preparation of a cosmetic and / or dermatological composition that is generally safe, non-toxic, and neither biologically nor otherwise undesirable, and that is acceptable for cosmetic or dermatological use.

[0017] For example, the cosmetic or dermatological composition comprises at least 0.1% by weight of said extract in relation to the total weight of the composition, preferably at least 0.5%. In another formulation, the composition comprises from 0.01 to 20% by weight of said extract in relation to the total weight of the composition, preferably from 1 to 10%.

[0018] The composition of the invention may be in the form of a cream, lotion, oil-in-water or water-in-oil emulsion, or gel, these compositions being more or less fluid or solid and having the appearance of a cream, lotion, milk, serum, ointment, gel, paste, mousse, or stick. The extract of the invention, and compositions incorporating said extract, have demonstrated numerous beneficial effects for the skin. The invention therefore relates to the cosmetic or dermatological uses of the extract and compositions incorporating the extract as an agent or product providing the following effects: anti-inflammatory, anti-irritant, immunomodulatory, soothing, anti-erythema, anti-eczema, and / or anti-redness;

[0019] - anti-aging, anti-wrinkle, firming, plumping, lifting, resurfacing, smoothing and / or regenerating; and / or

[0020] - probiotic, antioxidant, moisturizing, healing, texturizing, brightening, illuminating and / or anti-fatigue.

[0021] Similarly, the invention relates to the use of the fermented extract according to the invention for the manufacture of a composition intended to prevent or treat:

[0022] - skin inflammation, including dermatitis, erythema, irritation, rosacea and / or eczema, and / or to promote healing and skin regeneration, and / or intended to prevent;

[0023] - the signs of skin aging, notably by stimulating fibroblast proliferation, collagen synthesis and / or improving skin elasticity and firmness, and / or

[0024] - cutaneous oxidative stress, to protect the skin against external aggressions, to modulate the cutaneous microbiota and / or to improve the hydration and barrier function of the skin.

[0025] Finally, the invention also relates to a cosmetic skin treatment process by applying to the skin an extract or composition according to the invention, and intended to obtain an anti-aging effect reducing the visible effects of skin aging, skin protection, antioxidant, soothing, anti-inflammatory, anti-irritation, immunomodulatory, calming, anti-redness, repairing, probiotic, antioxidant, moisturizing, healing, texturizing, brightening, illuminating and / or anti-fatigue.

[0026] Other features and advantages will become apparent from the detailed description of the invention, examples, and test results that follow.

[0027] LIST OF FIGURES

[0028] [Fig. 1] Characterization of the anti-lipoxygenase activity of fermented extracts of Nannochloropsis Oculata according to a production method: aqueous extract, 70% alcoholic extract, or extract from a fermentation a) predominantly alcoholic “MSSC”, b) predominantly alcoholic and acetic “MSIev”, or c) lactic “MSLP”. [Fig. 2] Assay of polyphenols in fermented extracts of microalgae according to a production method: Microalgae extracted with H2O, Microalgae extracted with 70% EtOH, or extract from a fermentation a) predominantly alcoholic and acetic “MSIev”, b) predominantly alcoholic “MSSC”, or c) lactic “MSLP”. Dosage by the Folin-Ciocalteu method (0.15 ml of 0.45pm filtered sample diluted 1 / 10 + 0.45ml Na2COs 4% + 0.45ml Folin1 / 10 heating at 70°C for 20' and reading OD 760nm).

[0029] [Fig.3] Effects of a fermented extract of Nannochloropsis obtained by dominant alcoholic fermentation on the in vitro release of pro-inflammatory cytokines by RAW264.7 macrophage cells. (p<0.001 vs. control, p<0.001, p<0.05 vs. LPS).

[0030] [Fig. 4] Images of the cell scraping test and quantification of the relative mobility of NIH / 3T3 cells after application of a fermented extract of Nannochloropsis Oculata obtained by dominant alcoholic fermentation.

[0031] [Fig. 5] Promoting effect of a fermented extract of Nannochloropsis obtained by dominant alcoholic fermentation on fibroblast proliferation and collagen synthesis.

[0032] [Fig. 6] Probiotic effects of a fermented Nannochloropsis extract obtained by predominantly alcoholic fermentation. Left: Growth curve of B. longum in the presence of a control (minimum growth medium) and the minimum growth medium + 2% MSCC3J11 fermented extract (upper curve). Center: Monitoring of growth by measuring OD in medium A (2% honey + 0.45 g / L NaHCO3), medium C (2% acacia fiber + 0.2% glucose), and 5% MSCC3 fermented extract + medium A. Right: Monitoring of Escherichia coli growth inhibition by measuring OD as a function of fermented extract concentration.

[0033] [Fig. 7] Change in facial parameters measured on 23 women aged 20 to 55 years after the use of a control cream containing allantoin and trehalose as active ingredients and a 2% cream of Nannochloropsis Oculata ferment produced by predominantly alcoholic fermentation. The testing instruments used were VisioFace WOOD, ElastiMeter, C-CUBE, Mexameter® MX 18, and Skin-Glossymeter GL 200. Test procedure: After cleansing the face and allowing it to rest at a constant temperature (20°C–25°C) and constant humidity (40%–60%) for 20 minutes, the scientists collected the image of the face on day 0 and tested skin elasticity, melanin, erythema, and gloss. The volunteers applied the cream to their face (left and right side) twice a day, morning and evening, after cleansing.At 7 days, 14 days and 28 days respectively, scientists collected the image of the face and tested skin elasticity, melanin, erythema and radiance (data collected after cleansing the face for 20 minutes).

[0034] [Fig. 8] Continuation of results from figure 7.

[0035] [Fig. 9] Typical facial redness after the use of a control cream comprising allantoin and trehalose as active ingredients and a 2% cream of a Nannochloropsis Oculata ferment from a predominantly alcoholic fermentation.

[0036] DETAILED DESCRIPTION OF THE INVENTION

[0037] The present invention relates to a fermented microalgae extract, in particular a Nannochloropsis Oculata ferment exhibiting excellent anti-inflammatory, antioxidant, soothing, moisturizing, restorative, and anti-aging properties. Specifically, the invention provides a Nannochloropsis Oculata extract obtained through a fermentative extraction process that remarkably modifies the biological characteristics and properties of this microalga.

[0038] In the present invention, fermentative extraction consists of using Nannochloropsis Oculata biomass as a fermentation substrate for a yeast and / or a lactic or acetic acid bacterium. This fermentation process makes the microalga's biomolecules available for cosmetic and / or dermatological use without the need for extraction solvents. The invention thus provides cosmetic and dermatological products incorporating a Nannochloropsis ferment, particularly Nannochloropsis Oculata, into their composition.

[0039] The term Nannochloropsis Oculata ferment refers to a "fermented extract" comprising soluble fractions obtained from the fermentation of the liquid phase of the culture medium and / or components derived from the fermented biomass, i.e., the solid microalgal residues. In a preferred embodiment, the Nannochloropsis Oculata ferment is a concentrate of the fermentation medium containing the solid microalgal residues and the liquid phase of the fermentation. Optionally, the Nannochloropsis Oculata ferment also includes specific microorganisms, such as bacteria or yeasts that participated in the fermentation. Fermentation not only makes biomolecules of cosmetic and dermatological interest available but also exceptionally transforms and enriches the biochemical profile of this microalga.Table 1 below shows a comparison of the characteristics of a fermented extract with those of an aqueous extract and an alcoholic extract of Nannochloropsis Oculata. In the results below and in the figures in the description, the Nannochloropsis Oculata ferment is referred to as the fermented extract.

[0040] [Table 1] Characteristics of Nannochloropsis Oculata extracts according to a method of obtaining: fermented extract (fermentative extraction), aqueous extract and 70% alcoholic extract.

[0041] Sample Density (g / ml) Dry matter (%) Fermented extract 1.131 ± 0.042 1.808 ± 0.054

[0042] Extract H2O 1.007 ± 0.038 1.192 ± 0.067

[0043] EtOH extract 70% 1.003 ± 0.035 0.891 ± 0.042

[0044]

[0045] The results show that the density and dry matter content of the fermented extract are significantly increased. Fermentation of Nannochloropsis occulta therefore results in a notable change in its metabolism, leading to an increase in the synthesis of biomolecules. It was also observed that the fermented Nannochloropsis extract exhibits greater absorption of the UV spectrum than the aqueous and alcoholic extracts. The fermentation process alters the biochemical composition of Nannochloropsis occulta and promotes the production of active biomolecules.

[0046] Table 2 below shows the composition of three batches of Nannochloropsis Oculata fermented according to different fermentation methods. Specifically, Nannochloropsis biomass is used as a substrate for predominantly alcoholic fermentation, alcoholic and acetic fermentation, or predominantly lactic fermentation. These three fermentation methods all allow for modification of the characteristics of the resulting extracts. [Table 2] Characterization of Nannochloropsis Oculata fermented according to a fermentation process. Alcoholic and acetic fermentation “MSIev”, predominantly alcoholic fermentation “MSSC”, and lactic fermentation “MSLP”.

[0047] MSIev MSSC MSPL analyses

[0048] Brix 3.6 3.3 5.3

[0049] Dry Matter % 2.5 1.8 5.2

[0050] (1 hour at 100°C)

[0051] AT% (total acidity 1.2 0.74 1.07

[0052] Acid equivalent

[0053] acetic)

[0054] pH 3.2 3.8 3.2 Conductivity 6.48 6.30 6.36

[0055] mS / cm

[0056] Polyphenols g / l 0.36 0.48 0.42

[0057] (Acid Eq.)

[0058] (Gallic)

[0059] Reducing sugars <0.25% <0.25% 2.3%

[0060] totals % (Eq.

[0061] Sucrose)

[0062] Ethanol % 1.2 1.7 0.3

[0063] Anti-power 41.35 49.90 42.10

[0064] oxidant

[0065] % Inhibition 38 41 26

[0066]

[0067] lipoxygenase

[0068] All three extracts show higher density and dry matter compared to the aqueous and ethanolic extracts (see Table 1). However, each fermentation gives the extract a specific profile.

[0069] As can be seen in Figure 1, the extracts from dominant alcoholic fermentation (MsSC), alcoholic and acetic fermentation (MSIev), and lactic fermentation (MSPL) all exhibit high anti-lipoxygenase activity, significantly superior to that of an aqueous extract and a 70% ethanol extract of the microalga. The fermented extracts according to the invention therefore possess remarkable anti-inflammatory potential.

[0070] Extracts obtained by predominantly alcoholic fermentation (MsSC) and lactic acid fermentation (MSPL) also allow for an increase in the polyphenol concentration of the extract, particularly compared to an aqueous extract and an extract fermented by alcoholic and acetic fermentation (see Figure 2). In a preferred embodiment, the invention relates to an extract of Nannochloropsis Oculata obtained by predominantly alcoholic fermentation. Figures 3 to 9 show the results of the characterization of the biological activity of an extract thus obtained, and / or of a cosmetic cream incorporating said extract.

[0071] The results in Figure 3 show that the fermented extract significantly reduces the production of pro-inflammatory cytokines such as NO, IL-6, TNF-α, and CXC / L in response to LPS stimulation. This anti-inflammatory activity is particularly relevant for cosmetic applications aimed at soothing skin prone to redness, irritation, and chronic inflammation such as rosacea or eczema.

[0072] The results in Figure 4 indicate that the fermented extract of Nannochloropsis promotes cell migration by increasing the relative mobility of NIH / 3T3 fibroblasts after 24 hours of treatment. This stimulation of cell migration is a key indicator of skin regeneration and wound healing. The results in Figure 5 also demonstrate that the fermented extract significantly stimulates fibroblast proliferation after 24 and 48 hours of exposure. Therefore, this extract can be used in cosmetic formulations designed to repair damaged tissue, accelerate the healing of superficial wounds, and contribute to cell renewal. These properties make it an ideal ingredient for anti-aging skincare, post-dermatological treatment products, and products intended for fragile skin.Furthermore, the increase in collagen content after 48 hours of treatment suggests that this extract promotes collagen synthesis, a key element in maintaining skin elasticity and firmness. Incorporating this extract into cosmetic products allows for the prevention and reduction of signs of aging by improving skin tone and density.

[0073] The probiotic effects of the fermented extract of Nannochloropsis Oculata have also been demonstrated. Figure 6 shows the significant increase in the growth of Bifidobacterium longum and Staphylococcus epidermidis in the presence of this extract, and a greater probiotic effect compared to the use of a probiotic acacia fiber medium (medium C). The extract therefore has beneficial potential for the skin microbiota. A balanced microbiota is essential for maintaining healthy skin, strengthening the skin barrier, and reducing irritation and inflammation. In parallel, the inhibition of Escherichia coli observed in this study underscores the antimicrobial properties of the fermented extract. This ability to modulate the microbiota can be exploited in cosmetic formulations for skin prone to microbial imbalances, blemishes, and redness, promoting a healthier skin environment.

[0074] The invention therefore proposes the integration of this extract into cosmetic care to improve the balance of the skin microbiota, but also to offer protection against external aggressions while strengthening the skin's barrier function.

[0075] The cosmetic properties of the fermented extract were also verified after its incorporation into a cosmetic cream. The results in Figures 7 and 8 show that the application of a cream containing 2% Nannochloropsis Oculata ferment led to a significant improvement in the evaluated skin parameters. Compared to the control, the depth, volume, and surface area of ​​wrinkles decreased more markedly with the fermented extract. Furthermore, the wrinkle surface area showed a greater reduction, confirming a smoothing and firming effect. In addition, skin elasticity increased considerably after 7, 14, and 28 days of treatment, suggesting a notable improvement in skin tone and firmness. These results indicate that the fermented extract possesses anti-aging properties that contribute to reducing the visible signs of aging and improving overall skin texture.The results in Figure 7 also show a significant improvement in skin texture, as well as a reduction in melanin levels and erythema, suggesting a brightening and soothing effect. The progressive increase in skin radiance throughout the study reinforces the potential of this extract for illuminating and anti-fatigue skincare.

[0076] The integration of this extract into anti-aging cosmetic formulations thus makes it possible to offer a tangible benefit in terms of the prevention and correction of wrinkles, by strengthening hydration and promoting a lasting tightening effect, resulting in a more even complexion, a reduction in redness and an overall improvement in skin quality.

[0077] Example of preparation of a fermented extract of Nannochloropsis Oculata by extractive fermentationa) Obtaining the biomass of Nannochloropsis Oculata

[0078] Nannochloropsis Oculata is cultivated in a conventional manner to produce biomass, which is then filtered and dried.

[0079] b) Fermentation extraction

[0080] A culture medium is prepared by dissolving 50 g / L of powdered Nannochloropsis Oculata biomass with 50 g / L of sugar at 50°C. The culture medium is inoculated with a starter culture at 30°C and incubated at 26°C for 1 to 12 days, specifically 6 days for the tests shown in the figures, except for the 11-day test in Figure 6. At the end of fermentation, the culture undergoes sterile filtration. This yields a fermented Nannochloropsis Oculata extract according to the invention. The extracts thus obtained are homogenized and characterized (Tables 1 and 2). The extract is preferably concentrated under vacuum and stabilized by freeze-drying or controlled drying for use as an active ingredient in a cosmetic and / or dermatological formulation.

[0081] In one embodiment, fermentation is predominantly alcoholic and the starter culture contains a yeast, chosen particularly from the genus Saccharomyces. Preferably, fermentation is carried out under anaerobic conditions. Under these conditions, an evolution of the microorganisms in the culture medium has been observed, notably a decrease in yeast over time.

[0082] In another embodiment, the fermentation is lactic and involves inoculating the medium with lactic acid bacteria of the genus Lactobacillus. In a third embodiment, the fermentation is a co-fermentation of alcohol and acetic acid carried out with Saccharomyces yeasts and Acetobacter acetic acid bacteria under controlled oxygenation. This fermentation method promotes the transformation of carbon substrates into bioactive compounds with enhanced regenerative activity and anti-aging properties.

[0083] In a preferred embodiment, and regardless of the type of fermentation performed, the fermentation is carried out for at least 5 days, preferably at least 8 days, to improve the properties of the biomass and the extract. Indeed, reproducibility tests have shown that a prolonged fermentation time increases the quality of the extract. The Nannochloropsis Oculata ferment thus obtained exhibits biological properties suitable for dermatological and cosmetic applications. It is intended to be formulated as serums, creams, or lotions to improve skin elasticity and firmness, reduce redness, decrease inflammatory skin reactions, promote cell regeneration, and strengthen the skin barrier to limit transepidermal water loss.Thus, the fermented extract of Nannochloropsis obtained according to the process described in the present invention constitutes an innovative and effective ingredient for the formulation of advanced dermatological care.

Claims

DEMANDS 1. Fermented microalgae extract obtained by extractive fermentation characterized in that it comprises a Nannochloropsis ferment, preferably Nannochloropsis Oculata.

2. Fermented extract according to claim 1, wherein said ferment is obtained from a predominantly alcoholic fermentation, a lactic fermentation, or an alcoholic and acetic fermentation of Nannochloropsis Oculata.

3. Fermented extract according to claim 1 or 2, wherein the extract has at least one of the following characteristics: - a dry matter content w / w greater than 1.4%, preferably greater than 1.6%; -a pH between 3 and 4; - a polyphenol content of at least 0.30 g / L, said polyphenols being calculated as gallic acid equivalents.

4. Composition comprising in a cosmetically or dermatologically acceptable medium an extract according to one of claims 1 to 3, and preferably comprising at least one adjuvant selected from the group consisting of antioxidants, emollients, moisturizing agents, anti-aging agents, preservatives, diluting and / or solubilizing agents, perfumes and mixtures thereof.

5. Composition according to claim 4 comprising at least 0.1% by weight of the fermented extract on the total weight of the composition, preferably at least 0.5%.

6. Cosmetic composition according to claim 4, in the form of a cream, lotion, oil-in-water or water-in-oil emulsion, gel, these compositions being able to be more or less fluid or solid and having the appearance of a cream, lotion, milk, serum, ointment, gel, paste, mousse or stick.

7. Fermented extract according to any one of claims 1 to 3, or composition according to any one of claims 4 to 6, for use as an anti-inflammatory, anti-irritant, immunomodulatory, soothing, anti-erythema, regenerating, skin-protecting and / or anti-redness agent.

8. Cosmetic or dermatological, and not therapeutic, use of an extract according to any one of claims 1 to 3, or of a composition according to any one of claims 4 to 6, as an anti-aging, anti-wrinkle, firming, plumping, and / or smoothing agent or product.

9. Cosmetic or dermatological, and not therapeutic, use of an extract according to any one of claims 1 to 3, or of a composition according to any one of claims 4 to 6 as an antioxidant, moisturizing agent or product.

10. Fermented extract according to any one of claims 1 to 3, or composition according to any one of claims 4 to 6 for its use as a probiotic and / or symbiotic agent.

11. Fermented extract according to any one of claims 1 to 3, or composition according to any one of claims 4 to 6 for its use as an anti-eczema and / or healing agent.

12. Fermented extract according to any one of claims 1 to 3, or composition according to any one of claims 4 to 6 as a brightening, illuminating and / or anti-fatigue agent or product.

13. Fermented extract according to any one of claims 1 to 3, or compositions according to any one of claims 4 to 6 as a lifting, texturizing and / or resurfacing agent or product.

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