Compositions containing loofah root extract

Abstract

The present invention relates to a root extract of the loofah (Luffa cylindrica) plant, enriched in bryonolic acid, the bryonolic acid representing at least 10% by weight relative to the total weight of the dry extract. The present invention also relates to a composition comprising the root extract.

Description

[Technical Field]

[0001] The present invention relates to a root extract of the plant Luffa cylindrica enriched with bryonolic acid, a method for producing the enriched root extract, and a cosmetic, dermatological, or nutraceutical composition containing the enriched root extract as an active ingredient. Another object of the present invention is the use of the enriched root extract or a composition containing the enriched root extract as an active ingredient to restore dysregulated bioenergetic functions (stimulate skin energy metabolism), restore cell viability even in exhausted cells, prevent stress induced by toxic agents, prevent oxidative stress, and more generally prevent and / or delay signs of aging, as well as to energize skin cells, and to control the expression of ectopic skin olfactory receptors (ORs) involved in immune response, skin inflammation, skin neurophysiology, skin metabolic regulation, wound healing, and skin stress response. [Background technology]

[0002] Skin is the first barrier of the human body. It protects organs from temperature and humidity differences and from external environmental attacks, such as UV rays or pollutant mediators. However, excessive chemical and physical stimuli (exposure to sunlight, light, or UV rays; stress and malnutrition) can impair the normal function of skin and induce its aging. Therefore, it is important to maintain skin regeneration and repair by stimulating the metabolism of the cells that compose it, and in particular cellular energy metabolism. Specifically, cells use energy to, among other things, manufacture new proteins, provide nutrients, eliminate cellular waste, and repair DNA damage.

[0003] The functionality of bioenergetic processes is crucial for maintaining good performance of all organs, including the skin. At the cellular level, three major metabolic pathways: glycolysis, the pentose phosphate pathway, and the tricarboxylic acid (TCA) cycle, are linked and constitute the first stage of cellular metabolism (Respiratory metabolism: glycolysis, the TCA cycle, and mitochondrial electron transport, Current Opinion in Plant Biology (2004)) (Non-Patent Document 1).

[0004] Glycolysis is a metabolic pathway that converts glucose into pyruvate and hydrogen ions. The free energy released in this process is used to form the high-energy molecules ATP (adenosine triphosphate) and NADH (reduced nicotinamide adenine dinucleotide). NADH plays a key role in energy metabolism by accepting and donating electrons. The TCA cycle, which occurs in the mitochondrial matrix, consists of many reactions that generate NADH and reduced flavin adenine dinucleotide (FADH2), which can then be used by the oxidative phosphorylation pathway to generate ATP. Like NADH, FADH2 is also considered a high-energy molecule for cellular metabolism. In fact, NADH and FADH2 can donate their electrons to electron carriers in the electron transport chain to drive oxidative phosphorylation, which is responsible for ATP production. Furthermore, the TCA cycle is central to many metabolic pathways, and therefore requires a constant supply of intermediates required for its activity. For example, oxaloacetate, which can be synthesized from pyruvate previously produced during glycolysis, is the direct precursor to the amino acid aspartate. This amino acid is important for protein synthesis. Pyruvate from glycolysis enters the TCA cycle through the action of the pyruvate dehydrogenase complex. The pentose phosphate pathway, which operates in the cytosol, oxidizes glucose to NADPH and other carbohydrates for biosynthesis.

[0005] As detailed above, these three pathways generate the metabolic products necessary for mitochondrial ATP production through the process of oxidative phosphorylation. Mitochondria are essential organelles at the heart of this process. The entire series of these energy conversions is called cellular respiration, and its good functioning ensures the health of skin cells.

[0006] Indeed, glycolysis takes place in the cytoplasm, while the TCA cycle occurs in the mitochondria. Therefore, solutes and metabolites derived from glycolysis must be internalized through the mitochondrial membrane to be passed on to the TCA cycle. This role is played by SLC25, a specific family of transporters whose purpose is to transport various metabolites back and forth across the mitochondrial membrane.

[0007] It has been demonstrated that defects in these SLC25 mitochondrial carriers, which supply mitochondria with inorganic phosphate and ADP, substrates for oxidative phosphorylation, are involved in diseases characterized by defective energy production (Diseases caused by defects of mitochondrial carriers: A review, Biochimica et Biophysica Acta, 2008) (Non-Patent Document 2). Mitochondrial transporters of the SLC25 family and associated diseases: a review, J Inherit Metab Dis. 2014 Jul;37(4):565-75. 2014) (Non-Patent Document 3). Among these, SLC25A12, a malate-aspartate shuttle, is described as responsible for transporting NADH produced during glycolysis into mitochondria to supply it to the TCA cycle.

[0008] Thus, SLC25A12 plays a critical role in linking glycolysis with the TCA and oxidative phosphorylation metabolic pathways.

[0009] The accumulation of environmental damage and stress leads to a depletion of energy metabolism, affecting overall metabolism (Reprogramming of energy metabolism as a driver of aging, Oncotarget. (2016)). Furthermore, mitochondrial dysfunction has been described in several diseases and is highly associated with aging (Lopez-Otin, et al. 2013; Daniel Schniertshauer, et al. Age-Dependent Loss of Mitochondrial Function in Epithelial Tissue Can Be Reversed by Coenzyme Q10, J Aging Res, 2018). This dysfunction is due to changes induced by exogenous harmful stress (UV radiation, pollution, etc.), but also by endogenous reactive oxygen species (ROS) produced during the oxidative phosphorylation process itself. In young, metabolically "healthy" cells, antioxidant systems are responsible for managing acceptable levels of ROS to maintain efficient oxygen respiration and ATP production in order to respond effectively to any threat (Bee Ling Tan et al. 2018). In aging cells, these systems are overwhelmed as a result of accumulating internal and external stresses.

[0010] As a result, the production of energy required for overall cellular metabolic function declines, leading to an imbalanced homeostasis and abnormal protein production program. This can have dramatic consequences associated with characteristic signs of aging skin. In particular, the extracellular matrix (ECM) is a complex arrangement of macromolecules synthesized by skin cells that provides skin with firmness and elasticity. These macromolecules are produced through the endoplasmic reticulum (ER) and then the Golgi apparatus, where they are synthesized and secreted after maturation. SLC35 is a specific family of transporters for nucleotide sugars and adenosine 3'-phospho-5'-phosphosulfate (PAPS) into the Golgi and ER, and thus plays an essential role in the maturation of matrix macromolecules (Molecular physiology and pathology of the nucleotide sugar transporter family (SLC35), Eur J Physiol, 2004) (Non-Patent Document 8).

[0011] Sponge gourd (also called loofah) (Luffa cylindrica, Cucurbitaceae) is an annual vegetable upland crop native to India and South Asia, distributed primarily in tropical to temperate regions. In China, various parts of the plant (including flowers, fruits, seeds, leaves, and roots) are used to relieve or alleviate inflammation, such as sore throats, rhinitis, mastitis, edema, swelling, burns, and hemorrhoids.

[0012] The pentacyclic triterpenoid bryonolic acid was first isolated from the roots of Bryonia dioica (Jacq.) (Cucurbitaceae) in 1960, and subsequently from the roots and cultured cells of Luffa cylindrica (Roem). Hi Jai Cho et al. (Formation of bryonolic acid in cucurbitaceous plants and their cell cultures, Phytochemistry, Vol. 31, No. 11, pp. 3893-3896, 1992) examined 21 species or varieties of Cucurbitaceous plants belonging to nine genera and their cell cultures for their ability to produce bryonolic acid. Bryonolic acid was found in all but two of the species examined and was located exclusively in the roots of mature plants or in the radicles of seedlings.

[0013] Several biological activities of bryonolic acid have been reported, including antiallergic properties in rodents and various cytotoxic and antitumor activities in multiple cancer cell lines ("Bryonolic acid production in hairy roots of Trichosanthes kirilowii Max. var. Japonica transformed with Agrobacterium rhizogenes and its cytotoxic activity" by Tadahiro Takeda et al. (1994) (Non-Patent Document 10)). In her doctoral thesis (The isolation and biological evaluation of anti-inflammatory and chemopreventive triterpenoid natural products, chapter 2, pp. 64-85, May 2015) (Non-Patent Document 11), Emily Clegg Barker demonstrated that bryonolic acid exhibits anti-inflammatory and antioxidant activities, as evidenced by its strong induction of heme oxygenase-1 expression.

[0014] Therefore, on the one hand, it is recognized that bryonolic acid exhibits interesting biological activities that can be valued, for example, in the cosmetics field. Furthermore, bryonolic acid is exclusively present in the roots of certain Cucurbitaceae species, such as Luffa, Cucumis, and Benincasa species, more particularly Luffa cylindrica. Therefore, to obtain this interesting compound, it is advantageous to have access to the roots of the plant. However, such an operation involves the destruction of all or part of the plant. Furthermore, the Nagoya Protocol and international standards (ISO 26000) provide guidelines for controlling access to biodiversity and ensuring sustainable development and social responsibility.

[0015] On the other hand, although the natural root extracts of the above-mentioned Cucurbitaceae species contain bryonolic acid, its concentration is not sufficient to produce significant and satisfactory results, especially in the cosmetic and / or dermatological fields.

[0016] JP-3659424B2 (Patent Document 1) describes a composition containing bryonolic acid derived from loofah root extract for use in wound healing, promoting cell proliferation, and promoting skin conditioning. JP-5946717B2 (Patent Document 2) discloses bryonolic acid extractable from loofah for promoting collagen type III production and wound healing in the skin. EP-0359196A2 (Patent Document 3) also discloses topical preparations containing loofah extract for moisturizing the skin, whitening, and removing and preventing wrinkle formation. JP-S62226912A2 (Patent Document 4) discloses a composition containing an extract of cultured induced roots of a Cucurbitaceae plant produced by infection with Agrobacterium. Such extracts can be used for moisturizing the skin or improving fine wrinkles, but the specification does not mention the route of application. Furthermore, JP-2019034899A2 (Patent Document 5) discloses a composition containing a loofah fruit and stem extract, free of bryonolic acid, for preventing wrinkle formation by inhibiting elastase activity as a route.

[0017] JP-H07109214A (Patent Document 6) relates to a sebum secretion promoter containing bryonolic acid derived from cultured cells of a Cucurbitaceae plant.

[0018] JP-2009256271A (Patent Document 7) provides an extract from the aerial parts of loofah to promote aquaporin expression and thus provide moisture-retaining function to the skin, and CN-107773470A (Patent Document 8) discloses a face mask based on loofah extract that can moisturize the skin.

[0019] JP-H11240823A (Patent Document 9) discloses a composition containing a loofah extract for promoting the growth of hair papillae.

[0020] JP-4716692B2 (Patent Document 10) discloses a composition based on an ethanol extract of loofah root that has excellent anti-inflammatory effects that suppress erythema UV.

[0021] KR-101256180B1 (Patent Document 11) discloses a skin whitening composition containing loofah seed extract as an active ingredient capable of inhibiting tyrosinase activity in melanocytes. [Prior art documents] [Patent documents]

[0022] [Patent Document 1] JP-3659424B2 [Patent Document 2] JP-5946717B2 [Patent Document 3] EP-0359196A2 [Patent Document 4] JP-S62226912A2 [Patent Document 5] JP-2019034899A2 [Patent Document 6] JP-H07109214A [Patent Document 7] JP-2009256271A [Patent Document 8] CN-107773470A [Patent Document 9] JP-H11240823A [Patent Document 10] JP-4716692B2 [Patent Document 11] KR-101256180B1 [Non-patent literature]

[0023] [Non-licensed document 1] Respiratory metabolism: glycolysis, the TCA cycle and mitochondrial electron transport, Current Opinion in Plant Biology (2004) [Non-licensed document 2] Diseases caused by defects of mitochondrial carriers: A review, Biochimica et Biophysica Acta, 2008 [Non-licensed document 3] Mitochondrial transporters of the SLC25 family and associated diseases: a review, J Inherit Metab Dis. 2014 Jul;37(4):565-75. 2014)

Non-licensed Document 4

Non-licensed Document 5

Non-licensed Document 6

Non-licensed Document 7

[0024] Despite the availability of root extracts of Cucurbitaceae species, such as Luffa, as active ingredients, there remains a need for novel cosmetic, dermatological or nutraceutical active ingredients consisting of improved root extracts of said plants enriched in bryonolic acid.Furthermore, such enriched root extracts are obtained in accordance with the principles of sustainable development, social and environmental value, and the significance of preserving plants.

[0025] There also remains a need for novel cosmetic, dermatological or nutraceutical compositions comprising at least a loofah (Luffa cylindrica) root extract enriched with bryonolic acid as an active ingredient, which advantageously allows the active ingredient to exhibit cosmetic or dermatological effects that have never been achieved or described before. [Means for solving the problem]

[0026] Surprisingly, it has been found that the amount of bryonolic acid in loofah roots can be increased, for example, by cultivating the plant under specific conditions. The inventors have demonstrated that cultivating the plant under specific conditions allows the amount of bryonolic acid in the roots to be enriched. The root extract enriched with bryonolic acid can be advantageously used as an active ingredient in the cosmetic, dermatological, and / or nutraceutical fields. Furthermore, the biological targets and pathways distinguished and reached by the enriched loofah root extract allow for comprehensive skin beneficial effects to be obtained compared to those described in the prior art. Indeed, it has been demonstrated that bryonolic acid-enriched loofah root extract can stimulate skin energy metabolism by modulating the mitochondrial transporter SLC25, upregulate the endogenous antioxidant system by stimulating glutathione S-transferase theta 2, glutathione peroxidases 2 and 3, detoxify peroxides by enhancing peroxiredoxin-2, efficiently repair cellular bioenergetic profiles altered by environmental stressors, improve skin structure, and / or reduce wrinkles and fine lines by stimulating collagen type I and type IV protein expression and elastic fibers.

[0027] The loofah root extract according to the invention and the loofah root extract obtained by the method according to the invention unexpectedly exhibit beneficial effects on the skin, in particular cell health and skin rejuvenation effects, based on the restoration of cellular and mitochondrial bioenergetic function while stimulating endogenous antioxidant properties to enhance overall cellular metabolism and ultimately stimulate extracellular matrix macromolecules. In other words, the loofah root extract according to the invention and the loofah root extract obtained by the method according to the invention are suitable, for example, for stimulating and / or restoring cutaneous energy metabolism, restoring cellular viability even in exhausted cells, inducing a protective response of the skin against external agents (toxic agents, sun or UV, light exposure, stress, malnutrition), preventing oxidative stress, and more generally for preventing and / or delaying the signs of aging, as well as for improving skin firmness, tone and elasticity.

[0028] Summary of the Invention The present invention relates to an extract of the roots of the plant Luffa cylindrica, which contains bryonolic acid, said bryonolic acid representing at least 10% by weight relative to the total weight of the dry extract.

[0029] The present invention also relates to a method for producing a loofah root extract, which comprises the steps of: a) cultivating loofah under soil-free conditions, in particular aeroponic cultivation; b) optionally stimulating the roots of said plants; c) solid / liquid extraction by root maceration of the roots, optionally stimulated in step b); d) recovering the extract obtained in step c), and e) Optionally, clarifying the extract recovered in step d) by dilution and / or successive filtration, in particular by clarification and / or sterile filtration.

[0030] The present invention also relates to a cosmetic and / or dermatological and / or nutraceutical composition comprising the loofah root extract as defined above and optionally one or more excipients, which are preferably cosmetically or dermatologically or nutraceutical acceptable.

[0031] The present invention also relates to an extract of the roots of the loofah plant as defined above, or a composition comprising an extract of the roots of the loofah plant as defined above, for use in stimulating and / or restoring skin energy metabolism, restoring cell viability (even in exhausted cells), inducing a protective response of the skin against external agents, preventing oxidative stress, preventing and / or delaying the signs of ageing, and / or improving the firmness, tonicity and / or elasticity of the skin.

[0032] The present invention also relates to the use of an extract of the roots of the loofah plant as defined above or a composition (preferably a cosmetic composition) comprising an extract of the roots of the loofah plant as defined above for stimulating and / or restoring skin energy metabolism, restoring cell viability (even in exhausted cells), inducing a protective response of the skin against external agents, preventing oxidative stress, preventing and / or delaying the signs of ageing and / or improving the firmness, tonicity and / or elasticity of the skin.

[0033] The present invention also relates to a cosmetic skin care method for preventing or delaying the appearance of skin ageing effects, which method comprises applying to at least a part of the body or facial skin a cosmetic composition as defined above.

[0034] All documents cited or referenced herein ("documents cited herein"), and all documents cited or referenced in the documents cited herein, together with any manufacturer's instructions, manuals, product specifications, and product sheets for anything mentioned herein or referenced in any document incorporated by reference herein, are hereby incorporated by reference herein and may be used in the practice of this invention. More specifically, all referenced documents are incorporated by reference to the same extent as if each individual document was specifically and individually indicated to be incorporated by reference. [Brief explanation of the drawings]

[0035] [Figure 1] Seahorse XF cell mitochondrial stress test profile (bioenergetic profile) [Figure 2] Figure 2 shows the mitochondrial stress profile of Seahorse XF cells (n=3) in 44-year-old fibroblasts in the presence or absence of a stressor (solvent) with or without a cosmetic composition containing a loofah root extract enriched with bryonolic acid according to the present invention. Respiratory profile. NT (control fibroblasts) for non-stressed cells. [Figure 3] Figure 3 shows skin morphological observations performed on formalin-fixed, paraffin-embedded skin sections stained with HES (hematoxylin, eosin, and saffron). Skin explant models were treated for 7 days with formulations containing 0 (placebo), 0.125%, 0.5%, and 1% of the bryonolic acid-enriched loofah root extract according to the present invention. Dermal densification is indicated by yellow staining. Scale bar: 100 μm. [Figure 4]Figure 4 shows elastic fiber staining performed on formalin-fixed, paraffin-embedded skin sections after modified Verhoeff's staining. Skin explant models were treated for 7 days with formulations containing 0 (placebo), 0.125%, 0.5%, and 1% of the bryonolic acid-enriched loofah root extract according to the present invention. Elastic fibers appear dark. Nuclei are stained blue. Scale bar: 100 μm. [Figure 5] Figure 5 shows the biomechanical properties measured with a cutometer on the facial skin of elderly volunteers after twice-daily application of a formulation containing 1% of the loofah root extract enriched with bryonolic acid according to the present invention or a placebo formulation: skin firmness (R0), skin tonicity (Ur), skin elasticity (Ue and R6). Results are expressed relative to day 0 and compared to the placebo group. Statistics: ANOVA with Dunnett's comparison test. ***: p<0.005, **: p<0.01, *: p<0.05.

[0036] Detailed Description of the Invention A first subject of the present invention relates to an extract of the roots of the Luffa cylindrica plant, which contains bryonolic acid, said extract representing at least 10% by weight relative to the total weight of the dry extract.

[0037] According to one embodiment of the invention, bryonolic acid represents at least 15% by weight, preferably at least 20% by weight, at least 25% by weight, at least 30% by weight, at least 35% by weight, at least 40% by weight, at least 45% by weight, at least 50% by weight, at least 55% by weight or at least 60% by weight, relative to the total weight of the dry extract.

[0038] According to one embodiment of the present invention, the root extract is in liquid form and contains a maceration solvent selected from isopropyl myristate, triglycerides, triethyl citrate, dicaprylyl ether, glyceryl isostearate, glyceryl stearate, ethyl acetate, vegetable oils, or mixtures thereof. Preferably, the solvent is dicaprylyl ether. More preferably, the maceration solvent is biobased dicaprylyl ether. Such a liquid root extract corresponds to a crude liquid extract obtained after step a) of cultivating loofah under soilless conditions, particularly aeroponic cultivation, optional step b) of stimulating the roots of the plant, step c) of optionally subjecting the roots stimulated in step b) to solid / liquid extraction by root maceration, and step d) of recovering the extract obtained in step c). Thus, the "maceration solvent" is the solvent used to obtain the root extract. Such a solvent must be selected from specific solvents to achieve the desired bryonolic acid content of at least 10% by weight based on the total weight of the dry extract.

[0039] According to one embodiment of the invention, the root extract may be in solid or sticky form after a further step e) of drying the root extract in liquid form, said drying being carried out according to any method known in the art for placing the root extract in liquid form in a hot, dry atmosphere, for example to evaporate the maceration solvent.

[0040] The solid or sticky root extract can be further diluted in a dilution solvent to obtain another type of liquid root extract. Thus, a "dilution solvent" is a solvent used to dilute the solid or sticky root extract obtained. The dilution solvent can be selected from alcohol, glycols, ethyl lactate, isopropyl myristate, triglycerides, triethyl citrate, dicaprylyl ether, glyceryl isostearate, glyceryl stearate, ethyl acetate, vegetable oils, or mixtures thereof. The alcohol is preferably selected from methanol and ethanol. In practice, to achieve the desired bryonolic acid content of at least 10% by weight based on the total weight of the dry extract, a specific solvent must be used as the maceration solvent, but other solvents may be used to dilute the solid or sticky root extract obtained before use. In particular, the dilution solvent does not need to be the same as the maceration solvent used for the obtained extract. In the context of the dilution solvent, the alcohol is preferably selected from methanol and ethanol. The glycol is preferably selected from pentylene glycol and glycerol. Preferably, the dilution solvent is selected from pentylene glycol and dicaprylyl ether. In a particular embodiment, the dicaprylyl ether is bio-based. More preferably, the dilution solvent is pentylene glycol. More preferably, the dilution solvent is dicaprylyl ether. Particularly preferably, the dilution solvent is bio-based dicaprylyl ether.

[0041] Depending on whether the root extract in liquid form is a crude liquid root extract in a maceration solvent or a dried root extract diluted in a dilution solvent, the root extract in liquid form may contain the maceration solvent or the dilution solvent.

[0042] "Plant root extract" means a product obtained by extraction of the enriched root of the plant Loofah, said root extract preferably being in liquid form, although not exclusively so.

[0043] "Bryonolic acid-enriched root" and "enriched root" refer to a root extract of a plant that contains a higher amount of bryonolic acid than a corresponding root extract of the same plant as found in nature. In other words, "bryonolic acid-enriched root extract" refers to a root extract that contains at least 10% by weight of bryonolic acid relative to the total weight of the dry extract.

[0044] "Bryonolic acid" means a pentacyclic triterpenoid compound having the following general formula (I):

[0045] [ka] "Dry extract" means a root extract of a plant that can be obtained by carrying out the method according to the invention, followed by a step of drying the extract obtained, said drying being carried out according to any method well known to those skilled in the art, in particular by placing the root extract in a hot, dry atmosphere or on a heated plate in order to evaporate the solvent.

[0046] The components of the present invention are described in more detail below. While these components are listed with specific embodiments, it should be understood that they can be combined in any manner and in any number to create additional embodiments. The various described examples and embodiments should not be construed as limiting the invention to only the explicitly described embodiments. This description should be understood to support and encompass embodiments combining the explicitly described embodiments with any number of the disclosed components. Furthermore, any permutation and combination of all components described in this application should be deemed to be disclosed by the description of this application, unless the context dictates otherwise.

[0047] Throughout this specification and the claims, unless the context requires otherwise, the words "comprise" and variations such as "comprises" and "comprising" are understood to mean the inclusion of a stated number, integer, or step, or group of numbers, integers, or steps, and not the exclusion of other numbers, integers, or steps, or group of numbers, integers, or steps. When used in the context of describing the invention (particularly in the context of the claims), the terms "a," "an," and "the" and similar referents should be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of individually referring to each separate value within that range. Unless otherwise indicated herein, each separate value is incorporated herein as if it were individually recited herein. All methods described herein can be performed in any suitable order, unless otherwise indicated herein or clearly contradicted by context. The use of any and all examples or exemplary phrases (e.g., "such as," "for example") provided herein is intended merely to provide a better understanding of the invention and does not limit the scope of the invention as otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0048] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Although the methods and materials described herein are preferred, other methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention.

[0049] Preferably, to obtain the loofah root extract according to the invention, when the roots are deemed sufficiently developed, they are contacted with an extraction solvent by maceration or, preferably, by maceration, followed by recovery and processing of the extraction solvent to extract therefrom the secondary metabolites, including bryonolic acid, released by the roots of the plant. This preferred method is adapted from the method developed by the company Plant Advanced Technologies (PAT) under the trademark "PAT Plantes à Traire®" and described in International Application WO-01 / 33942, the teachings of which are incorporated herein by reference.

[0050] A second subject of the present invention relates to a method for producing an extract of the roots of the plant Loofah, comprising the following steps: a) cultivating loofah under soil-free conditions, in particular aeroponic cultivation; b) optionally stimulating the roots of said plants; c) solid / liquid extraction by root maceration of the roots, optionally stimulated in step b); d) recovering the extract obtained in step c), and e) Optionally, clarifying the extract recovered in step d) by dilution and / or successive filtration, in particular by clarification and / or sterile filtration.

[0051] This method is a preferred method for producing the loofah root extract of the present invention. In a preferred embodiment of the present invention, the loofah root extract of the present invention is obtained by the method of the present invention. In a preferred embodiment of the present invention, the method of the present invention is used to produce the loofah root extract of the present invention.

[0052] In the context of the present invention, the term "cultivation under soil-free conditions" refers to any cultivation method in which plant root development does not occur in soil. More precisely, soil-free cultivation is a type of cultivation in which plant roots grow in a reconstituted medium separated from the soil. This cultivation medium is periodically irrigated with a known nutrient solution suitable for the plant being cultivated.

[0053] There are various soil-free cultivation techniques, such as substrate-free systems, which require an oxygen-enriched nutrient solution, and substrate-based systems. Substrate-free systems include aquaculture, in which the nutrient solution is contained in a growing tank without circulation; nutrient film technique (NFT), in which the nutrient solution is enriched with dissolved oxygen during its movement by exchange with air; and aeroponics, in which plant roots do not come into contact with either solid or liquid media. In fact, roots are nourished by a nutrient mist obtained by atomizing the nutrient solution (using a mister) in a closed-system medium. Substrate-based systems include subirrigation, in which the nutrient solution percolates through the substrate below, and percolation, in which the nutrient solution is distributed by discontinuous irrigation at the top of the system and then percolates to the bottom of the substrate. The inorganic or organic substrate is neutral and inert, such as sand, clay, or rock wool. The substrate may also be of synthetic origin.

[0054] The term "aeroponic plant cultivation" refers to a soil-free method of cultivation in which the plant roots are not in permanent contact with either a solid or liquid nutrient medium.

[0055] The term "nutrient solution" refers to a solution containing essential mineral salts (nitrogen - N, phosphorus - P, potassium - K) in optimal amounts and in optimal ratios to each other in order to obtain maximum root growth and maximum production of secondary metabolites.

[0056] The term "stimulating nutrient solution" refers to a nitrogen-deficient solution containing essential mineral salts (nitrogen - N, phosphorus - P, potassium - K) in optimal amounts to obtain maximum root growth and maximum production of secondary metabolites, especially bryonolic acid.

[0057] According to one embodiment of the method of the present invention, plants may be nourished using a nutrient solution mist obtained by atomizing a nutrient solution in a closed medium through a mister.

[0058] According to one embodiment of the method of the present invention, the plants can be placed with their aerial parts above the trays and their root parts below the trays, the trays being placed on a table forming a holding area for collecting excess liquid diffused towards the plants, and the trays being transported onto the table at various stations. The teachings of this technique suitable for aeroponic plant cultivation are described in more detail in International Application WO2018054704A1, which is incorporated herein by reference.

[0059] According to one embodiment of the method of the present invention, during step a), aeroponic plants are nutrient-supplemented by spraying the roots with a nutrient solution of essential mineral salts (nitrogen - N, phosphorus - P, potassium - K) to achieve maximum root growth and maximum concentration of secondary metabolites, including bryonolic acid, without harming plant survival. A person skilled in the art knows by common sense how to adapt the proportions and concentrations of various mineral salts to optimize plant growth, and in particular root growth. In this case, the mineral salt concentration of the nutrient solution is advantageously in the low conductivity range, ranging from 0.4 to 1.6 mS / cm, preferably from 0.8 to 1.2 mS / cm, to promote a greater diversity of secondary metabolites, including bryonolic acid, in the root extract.

[0060] Advantageously, the aeroponic cultivation conditions described above allow obtaining several secondary metabolites, including bryonolic acid, in amounts superior to those obtained in soil cultivation.

[0061] According to one embodiment of the method of the present invention, the method may include a root stimulation step. In this case, the root stimulation step b) of the plant comprises placing the roots in contact with a nitrogen-deficiency stimulation nutrient solution. In this case, the solution may contain a nitrogen ratio below that generally considered optimal for plant growth, especially root growth, advantageously less than 15% nitrogen, more advantageously no nitrogen, and the steps may be sequential or simultaneous.

[0062] In a preferred embodiment of the present invention, the method for producing a root extract of the plant Luffa comprises the following steps: a) cultivating loofah aeroponically; b) stimulating the roots of the plant; c) solid / liquid extraction of the roots stimulated in step b) by root maceration in a solvent; d) recovering the extract obtained in step c); e) Optionally, clarifying the extract recovered in step d) by dilution and / or successive filtration, in particular by clarification and / or sterile filtration.

[0063] According to one embodiment of the method of the present invention, root stimulation of the plant in step b) comprises contacting the roots with a nitrogen-deficient nutrient solution, which causes "nitrogen stress" that stimulates the production of secondary metabolites, in particular bryonolic acid.

[0064] According to a particular embodiment of the method of the present invention, the nitrogen deficiency stimulating nutrient solution is a solution containing less than 15% nitrogen, preferably less than 10% nitrogen, advantageously less than 8%, more advantageously less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1% nitrogen, and even more advantageously 0% nitrogen.

[0065] According to one embodiment of the method of the present invention, the root stimulation step b) significantly increases the content of secondary metabolites, including bryonolic acid, in the roots and thus promotes the flux of said metabolites from the roots into the selected solvent for extraction, making it possible to return the plant to cultivation and then reuse it without completely losing its viability. In other words, the plant stimulation step promotes the production yield and secretion of secondary metabolites, including bryonolic acid.

[0066] According to one embodiment of the method of the present invention, root stimulation in step b) is carried out by feeding the roots of the plant with a nitrogen-deficient N / P / K-stimulating nutrient solution vaporized or misted onto the roots.

[0067] According to one embodiment of the method of the present invention, step b) is carried out by spraying or macerating the roots of the plant with an N / P / K nutrient solution containing less than 6%, more preferably less than 3% nitrogen, said solution preferably being evaporated or misted onto the roots.

[0068] According to one embodiment of the method of the invention, step b) of stimulating the roots of the plants by feeding them with a nitrogen-deficient N / P / K nutrient solution vaporized on the roots is advantageously carried out for a period of between 10 days and 8 weeks, preferably 3 weeks.

[0069] According to one embodiment of the method of the present invention, during step b) the mineral salt concentration of the nutrient solution is in the low conductivity range, advantageously ranging from 0.4 to 1.2 mS / cm, preferably from 0.6 to 1.0 mS / cm, in order to promote a greater diversity of secondary metabolites, including bryonolic acid, in the root extract.

[0070] According to one embodiment of the method of the invention, step b) is carried out after step a).

[0071] According to one embodiment of the method of the present invention, steps b) and a) are carried out simultaneously, and then the nutrient solution is replaced by the stimulation solution.

[0072] Subsequently, the cultivated and stimulated plants are typically subjected to step c) of solid / liquid extraction by root maceration under predetermined conditions with respect to solvent, temperature, and extraction time, thereby ensuring the production of a root extract enriched in bryonolic acid along with other secondary metabolites. Indeed, the amount of certain fatty acids (palmitic acid, linoleic acid, and α-linolenic acid) also increases after thin-layer chromatography (TLC) analysis of stimulated loofah root extracts (data not shown). Solid / liquid extraction is a solvent-based extraction technique that involves extracting one or more chemical species that are solid and soluble in the solvent. Extraction results in the recovery of secondary metabolites (including bryonolic acid) contained in the roots into a suitable solvent by placing the solvent in contact with the roots for a suitable time (also known as a soaking step). The solvent containing the secondary metabolites, including bryonolic acid, released by the roots is then recovered.

[0073] According to a preferred embodiment of the method of the invention, step c) of solid / liquid extraction of the roots stimulated during step b) is preceded by an additional washing step in which the solvent diffused into the plant is clear water, thus limiting the supply of elements contained in the nutrient solution or in the stimulated nutrient solution to the extraction solvent during the step of immersion in the solvent.

[0074] According to one embodiment of the method of the present invention, root maceration is carried out on cut and dried roots.

[0075] According to another more specific embodiment of the method of the present invention, root maceration is carried out on the cut, then dried, and optionally pulverized roots. The drying method can be any suitable drying method known to those skilled in the art, in particular by subjecting the root biomass to a temperature of 30°C to 60°C for 24 to 72 hours, preferably in a dry environment. The root biomass can be dried in a ventilated oven in particular. The root biomass can be pulverized by any suitable pulverization method known to those skilled in the art, in particular by placing the root biomass in a ball mill, knife mill, or hammer mill. After the root cutting step, the plants are returned to aeroponic cultivation according to step a) and optionally b) to resume their root growth and promote the root production of secondary metabolites, preferably bryonolic acid.

[0076] According to another more particular embodiment of the method of the present invention, said method comprises the step of cutting the roots and drying the cut roots prior to the step of maceration, wherein maceration is carried out by placing the cut and dried roots in contact with a solvent.

[0077] According to another more particular embodiment of the method of the present invention, said method comprises, prior to the step of macerating said roots, the steps of cutting the roots, drying the cut roots, and subsequently grinding the cut and dried roots, wherein maceration is carried out by placing the cut, dried and ground roots in contact with a solvent.

[0078] According to a preferred embodiment of the method of the present invention, step c) of solid / liquid extraction by maceration of the roots comprises contacting the roots with a solvent selected from isopropyl myristate, triglycerides, triethyl citrate, dicaprylyl ether, glyceryl isostearate, glyceryl stearate, ethyl acetate, vegetable oils or mixtures thereof.

[0079] Preferably, the solvent can be selected from isopropyl myristate, triglycerides, triethyl citrate, dicaprylyl ether, glyceryl isostearate, glyceryl stearate, ethyl acetate and vegetable oil (e.g., apricot kernel oil, sunflower oil, rapeseed oil).In a preferred embodiment, the solvent is dicaprylyl ether.In a specific embodiment, the dicaprylyl ether is bio-based.More preferably, the solvent is bio-based dicaprylyl ether.

[0080] According to another preferred embodiment of the method of the present invention, the solid / liquid extraction step c) comprises placing the roots in contact, for example by maceration, with dicaprylyl ether, particularly preferably bio-based dicaprylyl ether.

[0081] According to one embodiment of the method of the present invention, step c) comprises leaving the roots in contact with the solvent, for example by maceration, for a time period between 30 minutes and 48 hours, more particularly between 1 hour and 4 hours.

[0082] According to another particular embodiment of the method of the invention, step c) comprises placing the roots in contact with the solvent, for example by maceration, at a temperature between 20°C (room temperature) and 80°C, more particularly between 40°C and 60°C.

[0083] According to another particular embodiment of the method of the invention, for root maceration of dried roots, the ratio of the amount of dry root to the amount of solvent ranges from 1 kg dry root / 10 kg solvent to 1 kg dry root / 100 kg solvent, more preferably from 1 kg dry root / 20 kg solvent to 1 kg dry root / 40 kg solvent.

[0084] In the context of the present invention, the term "root maceration in a solvent" preferably means maceration of roots for which the solvent placed in contact with the roots is a solvent which can be, for example, isopropyl myristate, triglycerides, triethyl citrate, dicaprylyl ether, glyceryl isostearate, glyceryl stearate, ethyl acetate and vegetable oils (e.g. apricot kernel oil, sunflower oil, rapeseed oil), more preferably said solvent is dicaprylyl ether.

[0085] According to one embodiment, the method according to the invention may advantageously comprise one or more additional steps of processing said plant root extract, which may be chosen from: -dilution, at least one filtration, in particular a clarifying filtration and / or a sterile filtration, -Solid / liquid extraction, -purification, -Bleaching liquid root extract.

[0086] Such processing methods are well known to those skilled in the art.

[0087] Another subject of the present invention relates to the extract of the roots of the plant Luffa cylindrica obtained by the method detailed above.

[0088] In the context of the present disclosure, the concentration of bryonolic acid in a root extract of a plant, such as loofah, is determined by measuring the area of ​​the peak corresponding to bryonolic acid on the chromatogram of the root extract analyzed by liquid chromatography coupled to mass spectrometry (UPLC-MS). The area of ​​the bryonolic acid peak is given by the relative area of ​​the peak in the chromatogram of the UPLC-MS analysis of a standard solution containing 125 mg / L of bryonolic acid. The quantification of bryonolic acid corresponds to:

[0089]

number

[0090] According to a preferred embodiment of the composition of the present invention, bryonolic acid represents at least 10% by weight, preferably at least 15% by weight, preferably at least 20% by weight, at least 25% by weight, at least 30% by weight, at least 35% by weight, at least 40% by weight, at least 45% by weight, at least 50% by weight, at least 55% by weight or at least 60% by weight relative to the total weight of the dry extract.

[0091] According to a preferred embodiment of the composition of the present invention, the extract of the roots of the loofah plant corresponds to 0.0001 to 15% by weight, preferably 0.001 to 10% by weight, more preferably 0.01 to 5% by weight, based on the total weight of the composition.

[0092] The mode of administration, dosage, and optimal formulation of the cosmetic composition according to the present invention can be determined according to criteria generally considered in establishing a cosmetic treatment suitable for the subject, such as skin type. Depending on the desired type of administration, the cosmetic composition according to the present invention may further comprise at least one cosmetically acceptable excipient. The cosmetic composition according to the present invention may further comprise at least one adjuvant known to those skilled in the art for cosmetic use, such as thickeners, preservatives, fragrances, dyes, chemical or mineral filters, moisturizers, thermal waters, etc.

[0093] The cosmetic composition according to the invention may further comprise other cosmetically active agents, such as other anti-aging agents or moisturizing agents, agents with calming, soothing or relaxing activity, agents stimulating skin microcirculation, sebo-regulating agents for the care of oily skin, cleansing or purifying agents, anti-radical agents, anti-inflammatory agents, chemical or inorganic sunscreens, etc.

[0094] Suitable cosmetic excipients are well known to those skilled in the art. For example, suitable cosmetic excipients may be selected from polymers, silicone compounds, surfactants, rheological agents, humectants, penetrants, oily ingredients, waxes, emulsifiers, film-forming agents, and fragrances, electrolytes, pH adjusters, antioxidants, preservatives, dyes, pearlescent agents, pigments, and mixtures thereof.

[0095] The cosmetic composition according to the invention is advantageously intended for topical application and may in particular be in the form of a cream, milk, lotion, gel, serum, spray, mousse, solution, ointment, emulsion, patch or mask.

[0096] The cosmetic composition according to the invention is intended to delay or repair the appearance of skin signs of aging (intrinsic or extrinsic), in particular of extrinsic aging, including firmness, elasticity, appearance and wrinkles. Advantageously, the composition of the invention makes it possible to stimulate and / or restore the energy metabolism of the skin, restore cell viability even in exhausted cells, induce a protective response of the skin against external agents (toxic agents, sun or UV, light exposure, stress, malnutrition), prevent oxidative stress and, more generally, prevent and / or delay the signs of aging, as well as improve the firmness, tonicity and elasticity of the skin.

[0097] Suitable dermatological excipients are also well known to those skilled in the art, for example, suitable dermatological excipients may be the same as those intended for use in cosmetics.

[0098] Suitable nutraceutical excipients are well known to those skilled in the art. Examples include water-soluble polymers such as cellulose-based polymers, acrylate polymers and copolymers, polyvinylpyrrolidone, water-soluble polyethylene glycol, vinyl copolymers, etc. The nutraceutical composition may be a food supplement containing the loofah root extract of the present invention. Typically, the nutraceutical composition may be in the form of a solid, coated or uncoated tablet, liquid, powder, soft or hard gelatin capsule, etc.

[0099] Another subject of the present invention relates to an extract of the roots of the loofah plant as defined above, or a composition comprising an extract of the roots of the loofah plant as defined above, for use in stimulating and / or restoring skin energy metabolism, restoring cell viability (even in exhausted cells), inducing a protective response of the skin against external agents, preventing oxidative stress, preventing and / or delaying the signs of ageing, and / or improving the firmness, tonicity and / or elasticity of the skin.

[0100] In particular, the root extract or the composition comprising the root extract is suitable for stimulating skin energy metabolism by modulating the mitochondrial transporter SLC25, upregulating the endogenous antioxidant system by stimulating glutathione S-transferase theta 2, glutathione peroxidases 2 and 3, detoxifying peroxides by enhancing peroxiredoxin-2, efficiently repairing cellular bioenergetic profiles altered by environmental stressors, improving skin structure, and / or reducing wrinkles and fine lines by stimulating collagen type I and type IV protein expression and elastic fibers.

[0101] Another subject of the present invention relates to the use of an extract of the roots of the loofah plant as defined above or a composition (preferably a cosmetic composition) comprising an extract of the roots of the loofah plant as defined above for stimulating and / or restoring skin energy metabolism, restoring cell viability (even in exhausted cells), inducing a protective response of the skin against external agents, preventing oxidative stress, preventing and / or delaying the signs of ageing and / or improving the firmness, tonicity and / or elasticity of the skin.

[0102] Another object of the present invention relates to a cosmetic skin care method for preventing or delaying the appearance of the effects of skin aging, which method comprises applying to at least a portion of the body or facial skin a cosmetic composition as defined above.

[0103] Olfactory receptors (ORs) are expressed and active in various human tissues, including the skin. Primarily localized in the olfactory epithelium and mediating the first step in odor recognition, ectopic ORs are expressed outside the nasal region throughout the body. These ectopic ORs may play a role in regulating physiological cellular functions far beyond their role in olfaction. Nevertheless, most of these physiological roles remain to be elucidated.

[0104] Transcriptome studies (data not shown) carried out on the loofah root extract as defined above demonstrated that different concentrations of said extract, as listed in Table 4 below, can subsequently regulate the transcription of several olfactory receptor gene families in human epidermis, such as OR1, OR2, OR4, OR5, OR8, OR9, OR10, OR11, OR51, OR52 and OR56, with most of the members of each family being described as being involved in several key physiological functions, such as immune response, inflammation, neurophysiology, metabolic regulation, wound healing and skin stress response. The loofah root extract according to the invention can therefore be used as a guardian of overall skin homeostasis.

[0105] Olfactory receptors OR2AT4, OR2A4 / 7, OR2AG2, OR6M1, OR6V1, OR5V1, OR10A6, OR11H4, OR11A1, and OR51B5 have been identified in human skin. For example, OR2AT4, which is regulated by several plant extracts, is involved in skin re-epithelialization, skin healing processes, and hair growth.

[0106] The root extract of the loofah plant, as defined above, has been shown to regulate the olfactory receptors OR10A6 and OR2AG2, which have been described as being involved in skin stress responses (Duroux et al., 2020). The extract also regulates OR2AG2, which may be involved in skin inflammation. Furthermore, downregulating the expression of OR2AG1 and OR1D2 in human keratinocytes, as demonstrated by using the extract for OR2AG2, may lead to the control of skin inflammation caused by external agents such as UV and pollutants. Furthermore, OR10G7 may be involved in inflammatory skin disorders, such as atopic dermatitis, because it has been shown to be overexpressed in undifferentiated keratinocytes isolated from atopic dermatitis. Furthermore, OR2AG1 and OR1D2 are also involved in airway inflammation, such as asthma, and other chronic lung diseases.

[0107] Since keratinocytes have immunoregulatory functions, upregulation of OR1N1 following metabolic activation, as demonstrated by the loofah root extract as defined above, may regulate skin immunity. Indeed, the extract was shown to induce overexpression of OR1N1.

[0108] It has also been shown that the above-defined loofah root extract regulates the expression of OR52B4, which is involved in the angiogenesis process.As a result, the extract may be able to control vascular diseases of the skin, such as rosacea, skin aging, and dark circles.Furthermore, the extract regulates human skin cell metabolism, which may involve OR8J1 and OR4D2.

[0109] OR1L4, OR4D11, and OR5B3 belong to the olfactory family of OR1, OR4, and OR5, respectively. Because the skin, as the olfactory epithelium, is a highly innervated tissue and is in direct contact with volatile particulate matter, these genes may be involved in stress response mechanisms. Interestingly, the root extract of the plant loofah (Luffa japonica), as defined above, was shown to regulate OR1L4 in a manner similar to that of particulate matter.

[0110] The loofah root extract, as defined above, also upregulates the expression of OR5P3, suggesting its role in skin neurophysiology. Indeed, it has been suggested that OR5P3 may be involved in neurogenesis and neuronal differentiation through its interaction with a peptide derived from nerve growth factor (VGF).

[0111] Therefore, another subject of the present invention relates to the use of an extract of the loofah root plant as defined above, or of a composition (preferably a cosmetic composition) comprising an extract of the loofah root plant as defined above, in the regulation of the expression of cutaneous ectopic olfactory receptors (ORs) selected from the family OR1, OR2, OR4, OR5, OR8, OR9, OR10, OR11, OR51, OR52 and OR56.

[0112] In particular, the regulation of skin ectopic olfactory receptors by said extract or by a composition comprising said extract may preferably be for use in: a) Activation of skin stress response mechanisms involving OR1L4, OR4D11, OR5B3, OR10A6, and OR2AG2; b) Regulation of skin inflammation caused by external agents, involving OR2AG2, OR1D2, and OR10G7; c) Improvement of OR52B4-related vascular diseases, especially rosacea, skin aging, and dark circles; d) metabolic cell regulation involving OR8J1 and OR4D2; e) Skin neurophysiology, particularly skin neurophysiology for skin neurogenesis and skin neuronal differentiation involving OR5P3.

[0113] However, the functional effects of the loofah root extract according to the present invention should not be construed as being limited to the descriptions set forth herein, and may be interpreted otherwise in the absence of any scientific contradiction. [Example]

[0114] example The following examples illustrate various embodiments of the present invention, but the present invention is not limited to these examples.

[0115] Example 1: Study of bryonolic acid content in different species Different species (listed in Table 1 and obtained from commercially available seeds) were selected based on their good root yield in the aeroponic system. Root samples were collected from plants grown in aeroponic conditions for one month without stimulation, with a defined nutrient solution of N / P / K composition equivalent to 15 / 10 / 30, to a conductivity of 0.4-1.6 mS / cm. The bryonolic acid content in each sample was analyzed according to the following protocol: After cutting, the roots were dried in a ventilated oven at 40 °C for 48 h. The dried roots were then ground in a ball mill. For each sample, 10 mg of dried and ground root powder was macerated in 1 ml of ethanol (100%) for 30 min with stirring at room temperature (20 °C). The extract to be analyzed was then filtered through a 0.2 μm syringe filter before injection into HPLC-UV-MS as described in Example 3 below.

[0116] [Table 1]

[0117] Luffa is the species that produces the most bryonolic acid (3-4 times higher) in its roots after one month in an aeroponic culture system without stimulation. This amount is also six times greater than the amount listed in Table 1 of Jai Cho et al. (Formation of bryonolic acid in cucurbitaceous plants and their cell cultures, Phytochemistry, Vol. 31, No. 11, pp. 3893-3896, 1992). According to the literature, this study confirms that luffa leaves do not contain bryonolic acid.

[0118] To confirm the concentration of bryonolic acid in roots grown under non-aeroponic conditions, loofah seedlings were grown in pots with soil in a greenhouse. The content of bryonolic acid in roots was analyzed according to the following protocol: the roots were cut, washed, and then dried in a ventilated oven at 40°C for 48 hours. The dried roots were then ground in a ball mill. For each sample, 12 mg of dried and ground root powder was macerated in 1.2 ml of ethanol (100%) for 60 minutes with stirring at room temperature (approximately 20°C). The extract to be analyzed was then centrifuged and the supernatant poured into a vial before injection into HPLC-UV-MS as described in Example 3 below. Bryonolic acid was detected in roots grown under non-aeroponic conditions, but the amount was below the HPLC limit of quantification. This result confirms the effect of aeroponic cultivation on improving the bryonolic acid content in loofah roots.

[0119] Example 2: Comparison of bryonolic acid content in root extracts from loofahs grown in an aeroponic system with or without a stimulation step Root extracts were prepared from dried roots of loofah obtained from commercial seeds (French supplier, Graines Baumaux). Seedlings were placed in aeroponic cultivation systems. Plants in aeroponic systems were cultivated for 4 weeks in a nutrient medium consisting of nitrogen, phosphorus, and potassium (N / P / K) in a ratio of 15 / 10 / 30, respectively, with a conductivity of 0.4-1.6 mS / cm. The roots were then cut (T0) and the plants were then transferred to two separate aeroponic systems (20 m 2 (surface area) to be placed: For non-stimulated conditions, the plants are maintained for 6 weeks in cultivation using a nutrient medium composed of 15 / 10 / 30 (N / P / K) with a conductivity of 0.4 to 1.6 mS / cm; - For the stimulation conditions, the plants are maintained in cultivation for 6 weeks using a nutrient medium composed of 0 / 15 / 40 (N / P / K) with a conductivity of 0.4 to 1.2 mS / cm.

[0120] A root sample is then taken, dried and crushed: this constitutes the first cutting.

[0121] After this first cutting, the plants are re-fed as follows: For non-stimulated conditions, the plants are maintained for 3 weeks in cultivation using a nutrient medium composed of 15 / 10 / 30 (N / P / K) with a conductivity of 0.4 to 1.6 mS / cm; - For the stimulation conditions, the plants are maintained for 3 weeks in cultivation using a nutrient medium composed of 0 / 15 / 40 (N / P / K) with a conductivity of 0.4 to 1.2 mS / cm.

[0122] The root sample is then taken again, dried and ground: this constitutes the second cutting.

[0123] The bryonolic acid content in each sample is analyzed according to the following protocol: After cutting, the roots are dried in a ventilated oven at 40°C for 48 hours. The dried roots are then ground in a knife mill. For each duplicate sample, 100 mg of dried and ground root powder is macerated in 1.4 ml of methanol (100%) for 30 minutes at room temperature (approximately 20°C) with stirring. The extract to be analyzed is then filtered through a 0.45 μm syringe filter before injection into the HPLC-UV-MS as described below.

[0124] The analytical instrument used was an Agilent 1200 HPLC system (1260 Infinity Quaternary Pump, 1200 Series HiP-ALS SL autosampler, 1260 Infinity Thermostatted Column Compartment, 1260 Infinity Diode Array Detector) operated in reverse phase using 150 mm x 3.0 mm, 2.7 μm Poroshell 120 (Agilent) C18 columns. The mobile phase consisted of solvent A (ultrapure Milli-Q water, Merck Millipore + 0.1% formic acid, Carlo Erba, Val-de-Reuil, France) and solvent B (acetonitrile, Sigma-Aldrich Chemie GmbH, Steinheim, Germany). The gradient was programmed as follows: phase B (%) 94% (0–14 min); 94–100% (14–15 min); 100% (15–20 min). The analytical flow rate is 0.4 mL / min at an oven temperature of 25 °C. At the column outlet, a diode array detector records the UV spectrum between 210 and 600 nm. This instrument is connected to a mass spectrometer (Agilent 6120 Quadrupole LC / MS) operated in negative mode with electrospray ionization (4.5 kV) and atmospheric pressure chemical ionization in the m / z range of 100 to 1000. The ChemStation software is used to run the system.

[0125] The bryonolic acid content and dry root weight are shown in Table 2 below.

[0126] [Table 2]

[0127] With stimulation, bryonolic acid production and root production (assessed by dry root weight) are improved compared to roots of the same plants in an aeroponic cultivation system without stimulation: - Increased yield of bryonolic acid in roots after stimulation (mg / g dry root): 7-fold increase with respect to the second cutting - Increased root production after stimulation (g / m 2 ): 1.5-fold increase for the second cleavage - Overall increased bryonolic acid yield (mg bryonolic acid / m 2 ): 11-fold increase

[0128] Example 3: Preparation of Luffa Root Extract by the Method of the Invention and Characterization of the Root Extract Obtained The root extract is prepared from dried roots of loofah obtained from commercially available seeds (e.g., from the French supplier, Graines Baumaux). The loofah root extract is obtained according to the following process: a) aeroponic cultivation of the loofah plant in a prescribed nutrient solution with N / P / K composition corresponding to 15 / 10 / 30 and conductivity of 0.4-1.6 mS / cm, respectively, for 1-6 weeks; b) stimulating the plants with nitrogen stress for 2 to 6 weeks by using a nutrient solution with an N / P / K composition containing less than 6% nitrogen, 15% phosphorus and 40% potassium and having a conductivity of 0.4 to 1.2 mS / cm; b') rinsing with clear water followed by draining of the roots stimulated during step b); b'') cutting the roots and drying them in a ventilated oven at a temperature of 30-60°C for 24-72 hours; c) solid / liquid extraction of dried loofah root by maceration in a solution of dicaprylyl ether at a temperature between 20°C and 80°C for 1 to 4 hours; d) recovering the root extract obtained during step c); e) clarifying the root extract recovered in step d) by clarification filtration.

[0129] The loofah root extract thus obtained has 840 mg of dry extract per kilogram of extract and contains 300±10 mg of bryonolic acid per kilogram of extract, corresponding to about 35% of bryonolic acid by weight of the dry extract.

[0130] Quantification of the amount of bryonolic acid in loofah root extracts was carried out following a liquid chromatography coupled with mass spectrometry approach.

[0131] The instrument used for the analytical steps was a Shimadzu Nexera X2 UPLC (LC-30AD pumps, SIL-30AC sample passer, CTO-20A oven, SPD-M20A diode array detectors; Kyoto, Japan) operated in reversed phase using a Kinetex® Biphenyl column (Phenomenex®, Torrance, CA, USA) with dimensions of 150 mm × 1.1 mm, 2.6 μm. The mobile phase consisted of solvent A (Ultrapure Milli-Q water, Merck Millipore + 0.1% formic acid, Carlo Erba, Val-de-Reuil, France) and solvent B (acetonitrile, Sigma-Aldrich Chemie GmbH, Steinheim, Germany). The gradient was programmed in the following manner: phase B (%) 5-75% (0-3 min); 75% (3-5.4 min); 75-90% (5.4-5.6 min), 90% (5.6-12 min), 90-5% (12-12.1 min), 5% (12.1-15 min). The analytical flow rate was 0.5 mL / min with an oven temperature of 40 °C. At the column outlet, a diode array detector recorded the UV spectrum between 190 and 370 nm. The instrument is connected to a mass spectrometer (Shimadzu LCMS-2020) operated in negative mode with electrospray ionization (4.5 kV) in the m / z range of 100 to 1000. The software LabSolutions is used to run the system.

[0132] Quantification of bryonolic acid is performed by measuring the area of ​​the peak of a bryonolic acid standard (bryonolic acid purity ≥ 98%; [CAS number: 24480-25-3]) prepared at a concentration of 125 mg / L in ethanol (100%).

[0133] The amount of bryonolic acid, expressed as mg of bryonolic acid per liter of extract, is calculated according to the following formula:

[0134]

number

[0135] Example 4: Comparison of bryonolic acid content in root extracts with the total weight of the dried extract Ten grams of dried loofah roots, cultivated for 6 to 18 weeks in a nutrient medium consisting of 0 / 15 / 40 (N / P / K) with a conductivity of 0.4 to 1.2 mS / cm, were extracted in 110 mL of ethyl lactate solvent at room temperature for 5 days with stirring. The mixture was filtered to obtain the root extract, and the bryonolic acid content and dry extract were measured. The bryonolic acid content was 5.9% by weight based on the total weight of the dry extract.

[0136] Ten grams of dried loofah roots, stimulated for 6 to 18 weeks in a nutrient medium consisting of 0 / 15 / 40 (N / P / K) with a conductivity of 0.4 to 1.2 mS / cm, were extracted in 110 mL of 1,3-propanediol solvent for 5 days with stirring at 60 °C. After the filtration step, the obtained filtered loofah root extract contained 3 wt% bryonolic acid relative to the total weight of the dried extract.

[0137] Ten grams of dried loofah roots, cultivated for 3 to 6 weeks in a nutrient medium consisting of 0 / 15 / 40 (N / P / K) with a conductivity of 0.4 to 1.2 mS / cm, were extracted in 250 mL of 100% ethanol at room temperature with stirring for 10 minutes to 5 hours. Different samples were collected at 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, and 5 hours. The different samples were filtered to obtain root extracts, and the content of bryonolic acid in these and the dry extract was determined. The maximum percentage of bryonolic acid obtained was 8.1% by weight relative to the total weight of the dry extract. The minimum percentage of bryonolic acid obtained was 5.7% by weight relative to the total weight of the dry extract.

[0138] After 3 to 6 weeks of incubation in a nutrient medium consisting of 0 / 15 / 40 (N / P / K) with a conductivity of 0.4 to 1.2 mS / cm, 1.2 g, 1.5 g, 2.0 g, or 3.0 g of dried loofah roots were extracted in 30 mL of 100% ethanol with stirring at room temperature for 30 minutes to produce four extracts with ratios of 40 g, 50 g, 66 g, and 100 g of dried root per liter of 100% ethanol. The root extracts were filtered, and the bryonolic acid content and dry extract content were determined. The maximum bryonolic acid content was 8.3 wt% of the total dry extract at 100 g of dried root per liter of ethanol. The minimum bryonolic acid content was 7.2 wt% at 40 g of dried root per liter of ethanol.

[0139] For the following examples, the loofah root extract enriched with bryonolic acid (hereinafter referred to as LCRE) corresponds to the product obtained according to Example 3, which is adjusted to 145 mg of bryonolic acid per kilogram of extract.

[0140] Example 5: LCRE stimulates gene expression of proteins involved in bioenergetic metabolic pathways and antioxidant metabolism (RHE model).

[0141] LCRE formulated at 0.125% and 0.5% as described in Table 4 was applied topically onto reconstructed human epidermis (RHE).

[0142] [Table 3]

[0143] The RHE tissues were then cultured at the air-liquid interface in Epilife medium (Fisher Scientific, M-EPI-500-A) containing supplements and antibiotics (Gentamycin, Fisher Scientific, 15710-049). They were maintained in a humid atmosphere at 37°C with 5% CO2. Topical treatment (2 mg / cm2) for 24 hours was performed. 2 After centrifugation, total RNA was extracted using the RNeasy Mini kit from Qiagen. Their concentration and integrity were analyzed by spectrophotometry and capillary electrophoresis. Transcriptome analysis was performed on the Affymetrix human Clariom S array according to the Affymetrix user manual. Genes statistically significantly modulated by LCRE are listed in Tables 5 and 6. Analysis was performed in comparison with a group treated with placebo gel cream.

[0144] LCRE up-regulates three major energy pathways: glycolysis, the pentose phosphate pathway (PPP), and the tricarboxylic acid cycle (TCA cycle, also known as the Krebs cycle), as demonstrated by overexpression of enzymes involved in these pathways, as shown in Table 5. LCRE confirms its ability to stimulate skin energy metabolism in this RHE model. In parallel, LCRE acted on the mitochondrial transporter SLC25, which shuttles various solutes across the mitochondrial membrane (Table 5). Table 5 shows the modulation of gene expression at 0.125% LRCE, while Table 6 shows the modulation at 0.5% LCRE.

[0145] [Table 4]

[0146] Modulation of gene expression analyzed by Affymetrix technology in RHE models treated with 0.125% LCRE. Abbreviations: PPP: pentose phosphate pathway, TCA cycle: tricarboxylic acid cycle or Krebs cycle, SLC25: mitochondrial transporter. All analyses are performed in triplicate. Statistical analysis was performed in comparison with placebo gel cream. Statistical test based on the moderated t" method performed in R Limma 3.26.8 (Smyth, 2004), p-value < 0.05.

[0147] [Table 5]

[0148] Modulation of gene expression analyzed by Affymetrix technology in RHE models treated with 0.5% LCRE. Abbreviations: PPP: pentose phosphate pathway, TCA cycle: tricarboxylic acid cycle or Krebs cycle, SLC25: mitochondrial transporter. All analyses are performed in triplicate. Statistical analysis was performed in comparison with placebo gel cream. Statistical tests were based on the moderated t" method performed in R Limma 3.26.8 (Smyth, 2004). ***: p<0.005, **: p<0.01 *, p<0.05, #: p<0.1

[0149] These solutes are primarily involved in the progression of the TCA cycle (Palmieri et al. 2014 and 2016). The results in Table 6 demonstrate the strong involvement of LCRE in mitochondrial metabolism. Furthermore, LCRE facilitates the exchange of solutes essential for glycolysis, PPP, and the TCA cycle. Interestingly, SLC25A12 was significantly modulated (p-value < 0.05, fold change 1.27). However, enhancing mitochondrial metabolism can result in oxidative stress. LCRE upregulates the endogenous antioxidant system. In particular, LCRE stimulates three enzymes: glutathione S-transferase theta 2, glutathione peroxidase 2, and glutathione peroxidase 3, which are responsible for protecting cells from oxidative damage by coupling reduced glutathione with harmful compounds to neutralize them. LCRE also enhances peroxiredoxin-2, which catalyzes the reduction of hydrogen peroxide and organic hydroperoxides to water and alcohols, respectively, and plays a role in cytoprotection against oxidative stress by detoxifying peroxides.

[0150] Example 6: LCRE stimulates the expression of proteins involved in bioenergetic metabolic pathways and antioxidant metabolism (skin explant model).

[0151] LCRE formulated at 0.125%, 0.5%, and 1% as described in Table 4 was applied topically to human skin explants. Skin explants were harvested after abdominal surgery from a female donor (42 years old, Caucasian, phototype III). Skin samples were stabilized in OxiProteomics® medium at 37°C and 5% CO2 for 24 hours, then 2 mg / cm2 was applied. 2The skin explants were treated with different formulations (a group treated with the LCRE formulation and a placebo group). A third control group was added. Topical application was repeated six times every 24 hours. The control group did not receive any treatment except for changing the medium every 24 hours. Eight hours after the last application, the skin explants were sampled. Half of each explant was used for protein analysis. Protein was extracted using lysis buffer. Protein concentration was determined using the Bradford method, and quality control of the extraction was confirmed by high-resolution SDS-PAGE. Mass spectrometry (MS) analysis was performed on a Dionex U3000 RSLC nano-LC system coupled to an Orbitrap Fusion mass spectrometer (Thermo Fisher Scientific).

[0152] It was observed that LCRE upregulates three major energy pathways (Tables 7 and 8): glycolysis, the pentose phosphate pathway (PPP), and the tricarboxylic acid cycle (TCA cycle, also known as the Krebs cycle), as indicated by the overexpression of enzymes involved in these pathways. In this skin explant model, LCRE confirms its ability to stimulate skin energy metabolism, as already demonstrated in Example 5 (and also in Example 7). Furthermore, as observed and confirmed in Example 7 below, LCRE exerts antioxidant properties. Indeed, overexpression of several antioxidant enzymes was observed.

[0153] Furthermore, in this skin explant model, the transcriptomic results were confirmed by mass spectrometry (Tables 7 and 8) and by immunofluorescence (data not shown), highlighting the stimulation of SLC25A12 by the formulation containing 0.125% LCRE, which subsequently plays a crucial role in the malate-aspartate shuttle across the mitochondrial membrane and plays an essential role in glycolysis, along with NADPH plus H. + (Palmieri, Eur J Physiol, 2004). SLC25A12 is also involved in regenerating the antioxidant capacity of glutathione.

[0154] [Table 6]

[0155] Protein expression was analyzed by mass spectrometry in a skin explant model treated for 7 days with a formulation containing 0.5% LCRE. Abbreviations: PPP: pentose phosphate pathway. All analyses were performed in triplicate. Statistical tests were based on binary comparisons to the placebo control group using t-test statistical analysis. **: p<0.01 *: p<0.05.

[0156] [Table 7]

[0157] Protein expression was analyzed by mass spectrometry in a skin explant model treated for 7 days with a formulation containing 1% LCRE. Abbreviations: PPP: pentose phosphate pathway; TCA cycle: tricarboxylic acid cycle or Krebs cycle; SLC25: mitochondrial transporter. All analyses were performed in triplicate. Statistical tests were based on binary comparisons against the placebo control group using t-test statistical analysis. **: p<0.01 *: p<0.05 Visualization of SLC25A12 expression and localization after topical application of the LCRE formulation (data not shown) was studied by immunofluorescence in formalin-fixed, paraffin-embedded skin sections. Heat-induced epitope retrieval (HIER) was performed using citrate buffer (pH 6.0), followed by anti-SLC24A12 rabbit polyclonal antibody (Atlas Antibodies) at 1 / 500, followed by an Alexa 568-conjugated secondary antibody. Nuclei were labeled with Hoechst 33342.

[0158] Example 7: LCRE restores dysregulated bioenergetic function

[0159] The bioenergetic profile of human dermal fibroblasts was determined using a Seahorse XF24 flux analyzer. The Seahorse XF24 allows the determination of several bioenergetic parameters by measuring the oxygen consumption rate (OCR): - Basal respiration is defined as the oxygen consumption used to satisfy the cellular ATP demand resulting from mitochondrial proton leak. This parameter indicates the cellular energy demand under baseline conditions, which is the difference between the initial rate and the minimum respiration obtained with the addition of rotenone and antimycin A. - The ATP production parameter indicates the ATP produced by mitochondria, which contributes to meeting the energy needs of the cell. This parameter is experimentally obtained by the reduction of the oxygen consumption rate upon injection of the ATP synthase inhibitor oligomycin, which blocks the F0 subunit of the ATP synthase and therefore reduces the respiration rate, which reflects ATP production. - The maximum respiration parameter indicates the maximum rate of respiration that cells can achieve. This parameter is obtained by adding to the experiment a proton-disintegrating uncoupler called FCCP, carbonyl cyanide-4(trifluoromethoxy)phenylhydrazone. FCCP mimics physiological energy demands by stimulating the respiratory chain to operate at maximum capacity, which causes the rapid oxidation of substrates (sugars, fats, and amino acids) to meet this metabolic challenge. - Spare respiratory capacity reflects the ability of cells to respond under increased energy demand or stress. This parameter can be an indicator of cellular fitness and flexibility. This parameter is defined as the difference between maximum and base respiration (Figure 1).

[0160] Normal human dermal fibroblasts (NHDFs) isolated from donor abdominal skin were cultured in DMEM medium containing 10% fetal bovine serum (FBS). Cells were seeded at a density of 15,000 cells / well and allowed to attach for 6 hours. The chemical stressor, 2-methyltetrahydrofuran, was either added directly to the culture medium at 9% (hereafter referred to as the stressor) or not (control condition). In this experiment, the stressor was used to mimic exogenous stress, as environmental damage and accumulated stress have been shown to lead to energy depletion and affect overall metabolism (Reprogramming of energy metabolism as a driver of aging, Oncotarget (2016)).

[0161] For treatment, LCRE was also added directly to the culture medium at concentrations of 0.5% and 1%. The cells were then cultured for an additional 18 hours, after which measurements were performed using the Agilent Seahorse XF Cell Mito Stress Test Kit according to the manufacturer's instructions. Protein quantification was performed for all experimental conditions to normalize OCR data. Normalization of OCR protein ensures accuracy and interpretation of the results. Indeed, in the experimental conditions, cells were exposed to the stressors, which only slightly affected cell viability. Total respiration profiles (normalized to protein content) are shown in Figure 2, and calculated bioenergetic parameters are listed in Table 9. Stressors suppressed cellular respiration in all experimental models. In the presence of stressors, a strong decrease or total suppression of four key bioenergetic parameters was observed: basal respiration, maximal respiration, ATP production, and spare respiratory capacity. This was expected and validated the stress model.

[0162] [Table 8]

[0163] Bioenergetic parameters calculated for normal human fibroblasts in basal (untreated cells), stressed (with 9% 2-methyltetrahydrofuran) and treated conditions (0.5 and 1% LCRE).

[0164] LCRE at 0.5 and 1% can efficiently restore cellular bioenergetic profiles altered by stressors. These results indicate that LCRE-treated cells respond effectively and can meet urgent energy demands under stress conditions (by increasing maximal respiratory parameters and spare respiratory capacity), resulting in the recovery of ATP production. 0.5% LCRE allows the recovery of respiratory function similar to control conditions, restoring identical respiratory levels in all conditions tested. Furthermore, 1% LCRE improves respiratory function, which becomes superior to control conditions (Figure 2).

[0165] Example 8: Anti-aging properties of LCRE

[0166] In the skin explant model described in Example 6, proteomic analysis revealed that LCRE possesses anti-aging properties, as evidenced by stimulation of collagen type I protein expression (fold change x2.21, p-value <0.05). Supplementary histological and immunohistological analyses were performed on these skin explants to highlight its anti-aging properties. Formalin-fixed, paraffin-embedded skin sections were stained with hematoxylin-eosin-saffron (HES), modified Verhoeff's staining, and immunofluorescence. HES staining allows for morphological observation of the skin (Figure 3), while modified Verhoeff's staining (Figure 4) allows visualization of elastic fiber content in the skin. Figure 3 demonstrates the dose-dependent anti-aging effect of LCRE compared to a placebo gel cream (Table 4). Indeed, dermal densification was observed after topical application of LCRE. Elastic fibers are known to be responsible for skin elasticity. Unfortunately, skin elasticity declines with age due to a slowdown in elastic fiber production and increased elastase activity (Imokawa et al., 2008). LCRE was demonstrated to promote elastic fiber production. Indeed, in Figure 4, dark staining increased dose-dependently with increasing LCRE concentration compared to the placebo gel cream (Table 4). Dark staining represents elastic fibers. Furthermore, type IV collagen was also analyzed. In fact, type IV collagen, a major component of the dermal-epidermal junction, is also known to be significantly degraded with age. The results again confirmed the anti-aging effect of LCRE, as a dose-dependent increase in type IV collagen expression was demonstrated after 7 consecutive days of topical application of LCRE.

[0167] In the RHE model described in Example 5, a preparation containing 0.5% LCRE was found to stimulate the expression of the SLC35 gene family (Table 10). SLC35 is also an NST nucleotide sugar transporter. NSTs are antiporters that not only transport nucleotide sugars and PAPS to the Golgi apparatus, but also transport reaction products back to the cytosol. They are involved in the protein maturation process (Song, Z. 2013, Ishida N., et al. 2004), suggesting that the extract aids in protein synthesis and maturation. Interestingly, both of these processes slow down during the aging process.

[0168] [Table 9]

[0169] Modulation of gene expression analyzed by Affymetrix technology in RHE models treated with 0.5% LCRE. Abbreviations: SLC35: endoplasmic reticulum and Golgi apparatus transporter. Statistical tests were based on the moderated t" method performed in R Limma 3.26.8 (Smyth, 2004). *: p<0.05 and #: p<0.1. All analyses were performed in triplicate.

[0170] Finally, the anti-aging properties of LCRE were evaluated in human volunteers during a double-blind and vehicle-controlled clinical trial. The recommendations of the Declaration of Helsinki and the International Conference on Harmonization Good Clinical Practice guidelines, as applicable to non-pharmaceutical studies, were adhered to. All volunteers provided written informed consent.

[0171] Clinical evaluation was performed under the supervision of a dermatologist by 20 Caucasian women (45-55 years old, mean age 51.8 years) who exhibited clinical signs of aging, including crow's feet (grade ≥2 according to R. Bazin and E. Doublet's Skin Aging Atlas, vol. 1, Caucasian type) and a lack of firmness and elasticity. Furthermore, a questionnaire confirmed that the volunteers enrolled in this study considered themselves stressed and tired. Furthermore, a questionnaire based on questions about self-confidence allowed the selection of nine volunteers who lacked self-confidence. This subselection made it possible to reliably identify whether a cosmetic composition containing LCRE, as described in Table 4, would help these nine volunteers gain self-confidence. The volunteers were asked to apply a cosmetic formulation containing 1% LCRE and a placebo gel cream twice daily to one side of their face for two months (Table 3). Biometric measurements were performed using a cutometer on days 0, 28, and 62. The cutometer allows the determination of the biomechanical properties of the skin by calculating specific parameters: skin firmness (R0 parameter), skin tonicity (Ur parameter), skin elasticity (Ue and R6 parameters). Illustrative facial images were then taken with VisiaCR®.

[0172] Referring to Figure 5, LCRE improves skin firmness, skin tension and skin elasticity. In fact, after 28 days and 62 days of topical application of LCRE, all parameters are statistically significantly increased. In contrast, placebo formulation does not produce any improvement.

[0173] Of the nine unsure volunteers enrolled in the study, 78% of them described their skin as more beautiful, renewed, and revitalized after 28 days of topical application of the LCRE-containing composition (compared to 56%, 67%, and 67%, respectively, for the placebo formulation).

[0174] Furthermore, an improvement in the depth of wrinkles on the skin was observed with the LCRE-containing composition, and the skin microrelief was also improved, making the skin appear smoother and firmer.

Claims

1. 1. An extract of the roots of the plant Luffa cylindrica, comprising bryonolic acid, said bryonolic acid representing at least 10% by weight relative to the total weight of the dry extract.

2. 2. The root extract of claim 1, wherein the bryonolic acid represents at least 15% by weight, preferably at least 20% by weight, at least 25% by weight, at least 30% by weight, at least 35% by weight, at least 40% by weight, at least 45% by weight, at least 50% by weight, at least 55% by weight or at least 60% by weight, based on the total weight of the dry extract.

3. in liquid form and: a) a maceration solvent selected from isopropyl myristate, triglycerides, triethyl citrate, dicaprylyl ether, glyceryl isostearate, glyceryl stearate, ethyl acetate, vegetable oils or mixtures thereof; or b) a diluent solvent selected from alcohols, glycols, ethyl lactate, isopropyl myristate, triglycerides, triethyl citrate, dicaprylyl ether, glyceryl isostearate, glyceryl stearate, ethyl acetate, vegetable oils or mixtures thereof; 3. The root extract of claim 1 or 2, comprising:

4. Steps below: a) cultivating loofah under soil-free conditions, in particular aeroponic; b) stimulating the roots of the plant; c) a step of solid / liquid extraction by root maceration of the roots stimulated in step b), said maceration comprising contacting the roots with a solvent selected from isopropyl myristate, triglycerides, triethyl citrate, dicaprylyl ether, glyceryl isostearate, glyceryl stearate, ethyl acetate, vegetable oils or mixtures thereof; d) recovering the extract obtained in step c), and e) optionally clarifying the extract recovered in step d) by dilution and / or successive filtration, in particular by clarification and / or sterile filtration, 2. A method for producing the loofah root extract of claim 1, comprising:

5. Steps below: a) cultivating loofah aeroponically; b) stimulating the roots of the plant; c) a step of solid / liquid extraction by root maceration in a solvent of the roots stimulated in step b), said maceration comprising contacting the roots with a solvent selected from isopropyl myristate, triglycerides, triethyl citrate, dicaprylyl ether, glyceryl isostearate, glyceryl stearate, ethyl acetate, vegetable oils or mixtures thereof; d) recovering the extract obtained in step c); e) optionally clarifying the extract recovered in step d) by dilution and / or successive filtration, in particular by clarification and / or sterile filtration, The method of claim 4, comprising:

6. 6. The method of claim 4 or 5, wherein step b) of root stimulation of the plant comprises contacting the roots with a nitrogen deficiency stimulation nutrient solution.

7. 7. The method of claim 6, wherein the nitrogen-deficient stimulating nutrient solution is a solution containing less than 15% nitrogen, preferably less than 10% nitrogen, less than 8%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, or 0% nitrogen.

8. 8. The method according to any one of claims 4 to 7, further comprising the step of cutting the roots and drying the cut roots prior to the maceration step.

9. A loofah root extract obtained by the method according to any one of claims 4 to 8.

10. A cosmetic or dermatological or nutraceutical composition comprising a loofah root extract as defined in any one of claims 1 to 3 or 9, and optionally one or more excipients, which are preferably cosmetically or dermatologically or nutraceutical acceptable.

11. 11. The composition according to claim 10, wherein said extract of the roots of the loofah plant represents from 0.0001 to 15% by weight, preferably from 0.001 to 10% by weight, more preferably from 0.01 to 5% by weight, relative to the total weight of the composition.

12. Use of an extract of the roots of the plant Luffa as defined in any one of claims 1 to 3 or 9, or of a composition (preferably a cosmetic composition) comprising an extract of the roots of the plant Luffa as defined in any one of claims 1 to 3 or 9, for stimulating and / or restoring skin energy metabolism, restoring cell viability (even in exhausted cells), inducing a protective response of the skin against external agents, preventing oxidative stress, preventing and / or delaying the signs of ageing, and / or improving skin firmness, tonicity and / or elasticity.

13. A cosmetic skin care method for preventing or delaying the appearance of skin aging effects, comprising applying to at least a portion of the body or facial skin a cosmetic composition as defined in claim 10 or 11.

14. 10. Use of an extract of the roots of the plant Luffa cylindrica as defined in any one of claims 1 to 3 or 9, or a composition (preferably a cosmetic composition) comprising an extract of the roots of the plant Luffa cylindrica as defined in any one of claims 1 to 3 or 9, in regulating the expression of cutaneous ectopic olfactory receptors (ORs) selected from the family OR1, OR2, OR4, OR5, OR8, OR9, OR10, OR11, OR51, OR52 and OR56.

15. 15. The use according to claim 14, wherein the use is for: a) activation of skin stress response mechanisms involving OR1L4, OR4D11, OR5B3, OR10A6 and OR2AG2; b) Regulation of skin inflammation caused by external agents involving OR2AG2, OR1D2, and OR10G7; c) Improvement of vascular diseases, especially rosacea, skin aging and dark circles, which are associated with OR52B4; d) metabolic cellular regulation involving OR8J1 and OR4D2; e) Cutaneous neurophysiology, particularly for cutaneous neurogenesis and cutaneous neuronal differentiation involving OR5P3.

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