Cosmetic composition comprising an extract of perilla

Abstract

The invention relates to an aqueous, aqueous-alcoholic or alcoholic extract of at least one plant or at of least one part of a plant of the Perilla genus, to the method for obtaining same, to a composition containing same, and to various uses thereof, in particular cosmetic uses.

Description

[0001] Cosmetic composition containing a plant extract

[0002] The invention relates to an aqueous, hydro-alcoholic or alcoholic extract of at least one plant or at least part of a plant of the genus Perilla, its method of obtaining it, a composition containing it, as well as various uses, including cosmetics.

[0003] The skin is a semi-permeable protective layer that prevents water loss, thus ensuring adequate hydration and protecting the body against the migration of microorganisms through the skin. Therefore, the epidermis acts as a moisture barrier, but also protects the body from the environment.

[0004] Nevertheless, the skin is subjected to numerous external aggressions that can lead to molecular processes detrimental to its structure, such as exogenous factors like solar radiation, air pollution, tobacco smoke, nutrition, and temperature, and endogenous factors like lack of sleep or stress. In particular, ultraviolet (UV) radiation is considered the primary cause of skin aging, as it induces reactive oxygen species (ROS), DNA mutations, and immunosuppression, which can lead to photoaging.

[0005] There is always a need for new agents, particularly in the cosmetic field, and especially those capable of preventing and / or combating skin aging, particularly photo-induced skin aging, improving cell longevity, restoring the skin barrier function and / or improving skin hydration.

[0006] The Applicant has found that an aqueous, hydro-alcoholic, or alcoholic extract of at least one plant or part of at least one plant of the genus Perilla exhibits interesting and unexpected activities related to strengthening the skin barrier function, skin cell longevity, and skin hydration, through the stimulation or inhibition of physiological mechanisms. Indeed, as demonstrated in the examples below, the extract of at least one plant or part of at least one plant of the genus Perilla according to the invention significantly stimulates biological pathways related to the skin barrier function, particularly markers of skin aging, and especially photoaging, such as ceramides, Aquaporin 3 (AQP3), Filaggrin, and Claudin 1, as well as significantly reducing lipid peroxidation.

[0007] The invention therefore relates, according to a first aspect, to an aqueous, hydroalcoholic or alcoholic extract of at least one plant or of at least a part of at least one plant of the genus Perilla, said extract comprising at least 2%, preferably at least 3%, in particular at least 4%, better at least 5%, and most particularly at least 6%, by weight of total polyphenols, and most particularly at least 0.2%, preferably at least 0.3%, and in particular at least 0.4%, by weight of rosmarinic acid, relative to the total dry weight of the extract.

[0008] Preferably, the extract according to the invention is substantially free of organic solvent.

[0009] For the purposes of this invention, "organic solvent" means all solvents containing organic compounds, such as methanol, ethanol, acetone, ethyl acetate, chloroform and dichloromethane.

[0010] By "substantially free" is meant that the extract according to the invention comprises a content less than or equal to 0.1% by weight, in particular less than or equal to 0.01% by weight relative to the total weight of the extract.

[0011] According to one embodiment, the extract according to the invention comprises less than 0.1% by weight, in particular less than 0.01% by weight, relative to the total weight of said extract, of organic solvent, in particular selected from the group consisting of methanol, ethanol, acetone, ethyl acetate, chloroform, dichloromethane and mixtures thereof.

[0012] According to a preferred embodiment, the extract according to the invention is free of organic solvent, and in particular free of methanol, ethanol, acetone, ethyl acetate, chloroform, and dichloromethane.

[0013] According to one embodiment, the extract according to the invention is an aqueous extract of at least one plant or at least part of at least one plant of the genus Perilla, said extract comprising at least 2%, preferably at least 3%, in particular at least 4%, better at least 5%, and most particularly at least 6%, by weight of total polyphenols, and most particularly at least 0.2%, preferably at least 0.3%, and in particular at least 0.4%, by weight of rosmarinic acid, relative to the total dry weight of the extract.

[0014] Preferably, the extract according to the invention is an aqueous extract of at least one plant or at least a part of at least one plant of the genus Perilla, said extract comprising at least 2%, preferably at least 3%, in particular at least 4%, better at least 5%, and most particularly at least 6%, by weight of total polyphenols, and most particularly at least 0.2%, preferably at least 0.3%, and in particular at least 0.4%, by weight of rosmarinic acid, relative to the total dry weight of the extract; and said extract being free from methanol, ethanol, acetone, ethyl acetate, chloroform, and dichloromethane. Preferably, the plant in question is of the species Perilla frutescens, also known as shiso.

[0015] The invention also relates, according to a second aspect, to a method for obtaining at least one extract according to the invention, characterized in that it comprises at least the following steps according to which:

[0016] (a) at least one plant or at least part of at least one plant of the genus Perilla is available, optionally in crushed and / or frozen form;

[0017] (b) the plant or plant part from step (a) is subjected to at least one extraction cycle comprising:

[0018] (b1) at least one microwave extraction step, optionally with stirring; and / or

[0019] (b2) at least one maceration extraction step, optionally under agitation;

[0020] preferably step (b1) and step (b2), when both present, being carried out within the same extraction device;

[0021] (c) Optionally, the extract obtained in step (b) is purified by filtration, ultrafiltration and / or nanofiltration; and

[0022] (d) Optionally, the extract obtained in step (b), or in step (c), is redispersed in a preservation solvent.

[0023] The invention also relates to an extract according to the invention that can be obtained by such a process.

[0024] Preferably, the extract according to the invention obtained by such a process is substantially free, or even preferably free, of organic solvent, in particular selected from methanol, ethanol, acetone, ethyl acetate, chloroform, dichloromethane and mixtures thereof.

[0025] The invention relates to a non-therapeutic use, in particular cosmetic, of at least one extract according to the invention, to prevent and / or combat skin aging, in particular photo-induced skin aging, improve cell longevity, restore the skin barrier function and / or improve skin hydration.

[0026] The invention further relates to a cosmetic composition, in particular a cosmetic, pharmaceutical, nutraceutical or food composition, and preferably a cosmetic composition, comprising at least one extract according to the invention or obtained by means of a process for obtaining the invention, optionally in association with at least one physiologically acceptable medium. Preferably, the extract or composition, in particular a cosmetic composition according to the invention, is administered (or adapted for application) orally or topically, and preferably topically.

[0027] The invention further relates to a non-therapeutic cosmetic treatment process, in particular for makeup and / or care, of a keratinous material, in particular of the skin, lips or hair, comprising at least one step of applying to said keratinous material at least one extract according to the invention or a composition according to the invention.

[0028] Plant of the genus Perilla

[0029] The raw material used in the present invention consists of at least one plant or at least part of at least one plant of the genus Perilla, which belongs to the Lamiaceae family. The Lamiaceae, or Labiatae, commonly known as mints, are a large family of dicotyledonous plants comprising approximately 6,000 species and nearly 210 genera. The Lamiaceae family includes many aromatic plants, notably mint, thyme, rosemary, lavender, sage, wild thyme, lemon balm, basil, oregano, savory, shiso, and marjoram.

[0030] Perilla is a botanical genus in the Lamiaceae family, which notably includes shiso (Perilla frutescens). Preferably, the plant or part of a plant of the genus Perilla used in the present invention is of the species Perilla frutescens. Perilla frutescens (also known as aromatic perilla or Nanking perilla), primarily called shiso in Japanese, zisu in Chinese, tîa tô in Vietnamese, or deulkkae in Korean, is a food, aromatic, medicinal, and ornamental plant. It has been cultivated and used in much of Asia and the Indian subcontinent since ancient times, and in Japan.

[0031] Extract extraction process

[0032] In particular, the extract according to the invention can be obtained by means of a process for obtaining at least one extract according to the invention, characterized in that it comprises at least the following steps according to which:

[0033] (a) at least one plant or at least part of at least one plant of the genus Perilla is available, optionally in crushed and / or frozen form;

[0034] (b) the plant or plant part from step (a) is subjected to at least one extraction cycle comprising:

[0035] (b1) at least one microwave extraction step, optionally with stirring; and / or (b2) at least one maceration extraction step, optionally with stirring;

[0036] preferably step (b1) and step (b2), when both present, being carried out within the same extraction device;

[0037] (c) Optionally, the extract obtained in step (b) is purified by filtration, ultrafiltration and / or nanofiltration; and

[0038] (d) Optionally, the extract obtained in step (b), or in step (c), is redispersed in a preservation solvent.

[0039] Preferably, a method of obtaining according to the invention does not include a concentration step and / or a purification step of the extract obtained in step (b), (c) or (d).

[0040] The advantageous and unexpected nature of an extract according to the invention derives in particular from step (a) of the process according to the invention and more particularly from optimized culture conditions, which in fact allow access to a plant or part of a plant, dedicated to be used for step (b) of extraction of the process of obtaining according to the invention, comprising optimized levels of total polyphenols and in particular of rosmarinic acid.

[0041] Thus, the plant or part of a plant in step (a) and dedicated to be used for step (b) of extraction of the process for obtaining according to the invention advantageously comprises:

[0042] - at least 2%, preferably at least 3%, in particular at least 4%, better at least 5%, and especially at least 6%, by weight of total polyphenols, and

[0043] - in particular, at least 1.5%, preferably at least 2%, in particular at least 2.5%, or even at least 2.75%, by weight of rosmarinic acid, relative to the total weight of the dry extract of said plant.

[0044] The advantageous and unexpected nature of an extract according to the invention further derives from step (b) of extraction, and in particular from the optimized extraction conditions identified by the inventors.

[0045] The advantageous and unexpected nature of an extract according to the invention finally stems from the fact that the process of obtaining it, after the extraction step (b), does not include a concentration step and / or a purification step generally implemented in the manufacture of plant extracts.

[0046] Such concentration and / or purification steps are well known to those skilled in the art. Examples include column chromatography, high-performance liquid chromatography (HPLC), liquid-liquid extraction, and precipitation. These concentration and / or purification steps involve solvents, generally organic solvents such as methanol, ethanol, acetone, ethyl acetate, chloroform, dichloromethane, and mixtures thereof, which are known to be hazardous and / or polluting.

[0047] The ability of a production process according to the invention to dispense with a concentration step and / or a purification step after the extraction step (b) results directly from the combination of the advantages inherent in steps (a) and (b) of the production process according to the invention.

[0048] The present invention is therefore particularly advantageous as it provides access to an extract containing satisfactory levels of total polyphenols, and in particular rosmarinic acid, from a production process which, after extraction step (b), is devoid of a concentration and / or purification step. The present invention is all the more advantageous as this absence of a concentration and / or purification step allows:

[0049] - a simplification of the production process,

[0050] - a reduction in the production costs of the extract,

[0051] - an improvement in the ecological impact of the production process,

[0052] - improved safety for operators, and

[0053] - access to an extract free from traces of concentration and / or purification solvents, and in particular free from traces of methanol, ethanol, acetone, ethyl acetate, chloroform, dichloromethane, and mixtures thereof, without prejudice to the concentration (or content) of total polyphenols, and particularly of rosmarinic acid.

[0054] By "free from any trace of concentration and / or purification solvents", it is understood that the extract according to the invention comprises a content less than or equal to 0.1% by weight, in particular less than or equal to 0.01% by weight, relative to the total weight of the extract, of concentration and / or purification solvents, and in particular free from any trace of methanol, ethanol, acetone, ethyl acetate, chloroform, dichloromethane, and mixtures thereof.

[0055] Preferably, the extract according to the invention is free of concentration and / or purification solvents, and in particular free of methanol, ethanol, acetone, ethyl acetate, chloroform, dichloromethane and mixtures thereof.

[0056] However, the prior art does not describe any plant extract or plant part of the genus Perilla, and particularly of the species Perilla frutescens, exhibiting similar quantities of polyphenols, and particularly of rosmarinic acid, obtained by an extraction process devoid of concentration and / or purification steps subsequent to the extraction step, and therefore free of hazardous and / or polluting organic solvents, even in trace amounts. Step (a)

[0057] The plant of the genus Perilla in step (a) is cultivated according to any method known to a person skilled in the art.

[0058] Preferably, the plant of the genus Perilla in step (a) is cultivated using a vertical cultivation method / device.

[0059] Preferably, the plant of the genus Perilla in step (a) is cultivated under controlled conditions, preferably in a cultivation unit ensuring control of growing conditions, in particular ensuring controlled regulation of water, humidity, temperature, and air quality and light supply, and in particular in which the plant of step (a) has been obtained by hydroponic or aquaponic culture.

[0060] By "controlled conditions", in the context of the present invention, we mean an enclosure (or "unit") in which at least the water and humidity parameters, the light, the temperature and the air quality are controlled and monitored, possibly by computer.

[0061] Thus, step (a) of a process for obtaining according to the invention is advantageously carried out from a plant of the genus Perilla cultivated in a cultivation unit ensuring total control of the growing conditions, in particular ensuring controlled regulation of water, humidity, temperature, and the level of air quality and light supply.

[0062] Regarding the water parameter, the water used for growing the plant can be chosen from any suitable water source, and in particular from municipal water, rainwater, or aquifer water. If necessary, the water can be treated before use, especially to remove any traces of contamination, particularly pesticides. Specifically, the water is purified using ultraviolet light, filters (preferably with 0.2-micron pore diameter), self-purifying microorganisms such as daphnia (also known as "water fleas"), and mixtures thereof.

[0063] Regarding the brightness setting, sunlight and / or artificial light can be used, particularly via LED technology.

[0064] Regarding the atmospheric parameter, the unit can be adapted to inject air of controlled quality, temperature and / or humidity.

[0065] Regarding the temperature parameter, a heating or air conditioning system can be used, possibly reversible.

[0066] The unit is also advantageously suited to operate in self-sufficiency, meaning that it produces the energy source(s) necessary for its operation. In particular, the energy used to power a unit according to the invention is clean energy (also called green energy), that is, a primary energy source that produces a low quantity of pollutants when transformed into final energy and then used as such. Preferably, the energy used to power a unit according to the invention is chosen from solar energy, including photovoltaic and thermal energy, wind energy, tidal energy, wave energy, hydroelectric energy, geothermal energy, space-based solar energy, nuclear energy, and biomass, preferably solar energy.

[0067] Thus, step (a) of a method for obtaining the product according to the invention is advantageously carried out using a plant of the genus Perilla obtained by hydroponic or aquaponic cultivation. By way of illustration, a suitable agricultural system is a hydroponic system as described, for example, in EP0010836, WO1996032835, W02016072042, and LIS2017049064. Another suitable agricultural system is an aquaponic system as described, for example, in WO2015160966 or W02008006172.

[0068] A person skilled in the art will know how to make the necessary adjustments to the unit to meet the "controlled conditions" criterion and ensure the cultivation of the plant.

[0069] A cultivation unit such as the one described above is advantageous in that it allows:

[0070] - to reduce, or even prevent, the risks of disease and / or contamination, - to limit the use of pesticides, or even to eliminate them entirely, and to avoid any contamination by pesticides from surrounding crops,

[0071] - year-round plant cultivation, with increased production yields because they are not subject to temperature variations, rain, hail, frost and / or predatory insects,

[0072] - remote control via computer,

[0073] - a reduction in the need for elements necessary for plant growth, - extraction from fresh plants, therefore rich in active ingredients of interest, and moreover in an undegraded form,

[0074] - the recycling of certain organic waste and excreta, particularly from cultivation and extraction stages, to contribute in whole or in part to the growth of future generations of plants, and

[0075] - to limit the use of costly transport and storage chains involving vehicles, machinery, fuels and packaging, implying a reduction in CO2 emissions and the need for energy and fossil carbon, and therefore a reduction in the impact on nature and costs. According to a particular embodiment, the plant of the genus Perilla in stage (a) is at a germinated seed stage (also called germination), microgreen or young shoot stage, generally grown without soil, and preferably microgreen.

[0076] A sprouted seed is a very early stage of maturity where the plant has a root system, but the cotyledon leaves are not yet developed or are very underdeveloped. The average growing time for most sprouted seeds is less than 10 days, and sometimes less than 5 days, from sowing to harvest. Sprouted seeds have superior properties, particularly nutritional ones, compared to dry seeds: vitamin and enzyme levels can be significantly increased during the germination process.

[0077] A germinated seed is therefore a plant that retains its root system and has an earlier stage of maturity than a microgreen.

[0078] A microgreen is a green vegetable harvested after germination. A microgreen has a single central stem that has been cut just above the soil line during harvesting (and therefore has no roots). It has fully developed cotyledon leaves and usually has a pair of very small, partially developed leaves. The average growing time for most microgreens is 10 to 14 days from sowing to harvest.

[0079] A young shoot is an early stage of maturity, but later than a germinated seed or microgreen. The average growing time for most young shoots is over 10 days, and often over 15 days, from sowing to harvest.

[0080] In general, sprouted seeds, microgreens, and young shoots contain considerably higher levels of vitamins and carotenoids than their mature plant counterparts.

[0081] The very short cultivation time, the very small size and the high concentration of active ingredient(s) make germinated seeds, microgreens and young shoots, preferably microgreens, types of plants perfectly suited to the present invention.

[0082] Thus, the extract according to the invention is obtained from at least one plant or at least part of at least one plant having a growth stage between 5 and 30 days, preferably between 7 and 25 days, and better between 10 and 22 days, or even between 17 and 22 days.

[0083] Preferably, the extract according to the invention is obtained from the roots of the plant, the aerial parts of the plant, and mixtures thereof, and preferably from the aerial parts of the plant.

[0084] Even more preferably, the extract according to the invention is obtained from the leaves, stems, flowers and flowering tops, buds, and mixtures thereof. Preferably, the extract according to the invention is obtained from at least one plant or at least part of at least one plant that has been cultivated:

[0085] - under irradiation at a light intensity between 100 and 600 pmol / cm² 2 / s, preferably between 150 and 500 pmol / cm 2 / s, and better between 200 and 400 pmol / cm 2 / s ;

[0086] - with a photoperiod in which exposure to light, whether continuous or discontinuous, is between 16 and 24 hours, preferably between 18 and 24 hours, better between 20 and 24 hours, and particularly between 22 and 24 hours; and / or

[0087] - under irradiation at a light intensity having a spectrum comprising between 70% and 40% of the flux in a wavelength between 400 nm and 500 nm, between 30% and 0% between 500 nm and 600 nm, between 50% and 10% between 600 nm and 700 nm and between 20% and 0% between 700 nm and 800 nm.

[0088] In a particularly preferred manner, the light intensity has a spectrum comprising between 60% and 50% of the flux in a wavelength between 400 nm and 500 nm, between 15% and 5% between 500 nm and 600 nm, between 30% and 20% between 600 nm and 700 nm and between 10% and 5% between 700 nm and 800 nm.

[0089] According to a particularly advantageous embodiment, the exposure to light is 24 hours. In other words, the extract according to the invention is obtained from at least one plant or at least part of at least one plant where the culture phase (i.e., the phase following the germination phase) has never experienced a period of darkness.

[0090] According to one embodiment, the culture phase can include several different photoperiods, and for example at least two photoperiods. Thus, by way of illustration, after the germination phase which generally takes place in darkness, a first culture phase can be carried out over a period of 5 days with a 16 / 8 photoperiod (i.e. 16 hours of light and 8 hours of darkness), followed by a second culture phase carried out over a period of 10 days with a 24 / 0 photoperiod (i.e. 24 hours of light and 0 hours of darkness).

[0091] In another variant, the light exposure period is continuous. For example, a 22-hour light exposure period would be completed in a single cycle.

[0092] According to yet another variant, the light exposure period is carried out discontinuously. For example, a 22-hour light exposure period will be achieved in at least two cycles, for instance, a first cycle of 10 hours and a second cycle of 12 hours, these two cycles being separated by a 2-hour darkness cycle. Preferably, the plant implemented in the present invention is cultivated with a photoperiod in which light exposure is achieved for at least 20 hours, preferably at least 22 hours, preferably continuously, and better still for 24 hours.

[0093] The growing temperature is advantageously between 20°C and 30°C, more advantageously between 22°C and 28°C, and better between 24°C and 26°C.

[0094] Step (b)

[0095] Step (b), in particular step (b1), can be carried out using any plant material from the whole plant, or from any part of said plant, such as roots, aerial parts of the plant, and mixtures thereof, and preferably leaves, stems, flowers and flowering tops, seeds, buds, roots, and mixtures thereof.

[0096] Preferably, extraction step (b), in particular step (b1), of a production process according to the invention is carried out from at least one plant or from at least a part of at least one plant:

[0097] - fresh, and preferably a frozen fresh plant; and / or

[0098] - previously dried, freeze-dried, frozen and / or ground, for example in the form of a powder smaller than 2 cm.

[0099] This configuration, using fresh or even frozen plants, advantageously reduces, or even prevents, a degradation in production yield and / or extract quality. Indeed, it is known that some active agents are difficult to preserve over time. This can be explained, in particular, by the development of biochemical mechanisms of tolerance to abiotic stresses, such as water and / or oxidative stress, which can lead to alteration or consumption of these active agents by the plant itself, or even to the production of unwanted molecules that may interact negatively with the active agent of interest. Now, for obvious reasons, the longer the time between harvesting the plant and extracting the active agent(s) it contains, the more the production yield and / or the quality of the active agent(s) can be affected.

[0100] For the purposes of this invention, "fresh" means a plant that has been harvested (for example, picked, pulled up, or extracted from the ground) less than 24 hours, preferably less than 12 hours, in particular less than 5 hours, better less than 3 hours, or even less than 1 hour, before step (b) of extraction.

[0101] For the purposes of this invention, "frozen" means a plant which, prior to step (b) of extraction, has undergone a freezing step, and therefore a plant which has been kept at a temperature below 0°C, preferably below 5°C, in particular below 10°C, or even below 15°C, preferably for at least 2 hours, in particular at least 5 hours, and better for a period of between 2 hours and 7 days, in particular between 5 hours and 5 days, better between 8 hours and 3 days, and especially between 12 hours and 1 day.

[0102] According to a first embodiment, a method of obtaining according to the invention does not include, in particular before step (b), a step of freezing, drying, dehydration, freeze-drying, grinding, decoction, enfleurage, infusion, addition of solvent(s), Soxhlet extraction, sonication-assisted extraction, supercritical fluid extraction, extraction based on the principle of sonoporation, and / or ultrasonic disintegration, and combinations thereof, of the plant or part of the plant of interest obtained in step (a).

[0103] According to another embodiment, a method of obtaining according to the invention may further comprise, in particular before step (b), at least one step of freezing, drying, dehydration, freeze-drying, grinding, decoction, enfleurage, infusion, addition of solvent(s), Soxhlet extraction, sonication-assisted extraction, supercritical fluid extraction, extraction based on the principle of sonoporation, and / or ultrasonic disintegration, and combinations thereof, of the plant or part of the plant of interest obtained in step (a).

[0104] According to a preferred embodiment, a method for obtaining the product according to the invention may advantageously include, before step (b):

[0105] - a freezing step of the plant or the part of the plant of interest. This variant is particularly advantageous when the extraction step is carried out in its native water; and / or

[0106] - a step of grinding the plant or the part of the plant of interest.

[0107] As previously stated, extraction step (b) is based on at least one extraction cycle, preferably comprising at least two successive different extraction steps (b1) and (b2), and preferably carried out within the same extraction device, or even the same receptacle, namely:

[0108] - a microwave extraction step, the product obtained from this first microwave extraction being designated as the "first aqueous extract"; and / or - a maceration extraction step applied to the plant or plant part of step (a) or to the first extract obtained in step (b1) (where present), the product obtained from this maceration extraction being designated interchangeably as the "extract of interest", "second extract" or "final extract". Step (b1) and / or step (b2) may be carried out without the addition of an extraction solvent or is / are carried out in the presence of at least one extraction solvent, identical or different, and preferably chosen from water, at least one alcohol, a hydro-alcoholic solvent, and mixtures thereof, and particularly chosen from glycerin, water, ethanol, isopropanol, propanediol, butylene glycol, and mixtures thereof.

[0109] Thus, according to a first embodiment, extraction step (b), and in particular steps (b1) and / or (b2), is / are carried out without the addition of an extraction solvent, using the plant's native water. In other words, no extraction solvent is used to carry out step (b), and in particular step (b1), and the extraction solvent used to carry out step (b2) is the native water extracted from the plant during step (b1).

[0110] Thus, the microwave extraction step (b 1) is in this variant of the embodiment carried out without the addition of extraction solvent, using only native water, namely water that is an integral part of the plant from the beginning, and in particular water that is an integral part of the plant cells.

[0111] According to a second embodiment, the extraction step (b1) is carried out in the presence of at least extraction solvent.

[0112] According to a particular embodiment, an extraction process according to the invention may comprise several extraction cycles, identical or different, in particular with respect to the power of the microwave extraction, the duration of step (b1) and / or (b2) and / or the respective sequences in step(s) (b1) and / or (b2), as described in more detail below.

[0113] Step (b1) - first microwave extraction step

[0114] The microwave extraction step can be referred to interchangeably as the "microwave radiation extraction step", and is carried out using a microwave radiation heat treatment device.

[0115] Such microwave heating technology is particularly suitable for plants because it allows direct heating to the core of the material for thin layers up to the order of a centimeter.

[0116] The microwave extraction device comprises at least one tank and at least one microwave generator with a frequency ranging, for example, but not limited to, between 300 MHz and 6000 MHz, and preferably around the standard frequencies of 2450 MHz or 915 MHz. This generator is connected, via at least one waveguide and flanges, to an enclosure. The purpose of the microwave generator(s) is / are to provide the energy necessary for the thermal treatment of the material being treated. Other heat sources may be added to increase the treatment efficiency, such as, for example, hot air or infrared radiation. The enclosure may have dimensions ranging, for example, but not limited to, between 300 mm and 2500 mm. One or more air inlet ports may be placed on the waveguides to allow, for example, dust removal or cooling.The flanges connecting the waveguide to the inlet waveguide can be equipped with a quick-release locking / unlocking system, for example, but not limited to, using compressed air. At least one window, for example, made of quartz or any other equivalent microwave-transparent material, can also be placed between the flanges to prevent dust from being drawn into the waveguide during heating cycles. In one embodiment, the enclosure is pivotally mounted, for example, on a base via an axis passing, for example, through the center of gravity of the enclosure. Pivoting means, such as a geared motor assembly, can allow the enclosure to be rapidly rotated about its axis. Such a device is described in particular in EP2530059.

[0117] In particular, the microwave extraction device includes a tilting rotating tank with ultrasound at the bottom of the tank.

[0118] Preferably, the microwave extraction step can be carried out:

[0119] - at a power between 600 W and 6000 W, preferably between 700 W and 5000 W, better between 800 W and 4000 W, and especially between 1000 W and 2000 W, or even between 2000 W and 6000 W (W = Watts);

[0120] - for a period of between 1 minute and 180 minutes, preferably between 2 minutes and 120 minutes, better between 5 minutes and 100 minutes, in particular between 10 minutes and 60 minutes, or even between 15 minutes and 30 minutes; and / or

[0121] - at a temperature between 30°C and 120°C, preferably between 40°C and 100°C, and better between 50°C and 90°C, preferably between 60°C and 80°C, and especially between 65°C and 75°C.

[0122] Preferably, the power used for the microwave extraction step is determined based on the plant mass. Thus, the microwave extraction step is advantageously carried out with a ratio of "plant mass (in grams) / power (in W)" between 0.5 and 2, and preferably with a ratio of 1:1. For example, for 600 grams of plant material, the power of the microwave extraction device is advantageously set at 600 W.

[0123] Preferably, the microwave extraction device may also include at least:

[0124] - a solution for stirring the plant to be treated; and / or - a rotating tank forming a cavity preferably made of a refractory metal with low resistivity, driven in rotation by means allowing its speed to be varied in order to optimize the stirring of the plant to be treated. Such an embodiment is described in particular in EP2530059.

[0125] Such embodiments are advantageous because they allow heat exchange between particles by convection, conduction, and, at higher temperatures, by radiation. This continuous exchange enables precise temperature balancing of the particles at every instant.

[0126] Advantageously, the microwave extraction step can be carried out via flash heat treatment, as described in FR2976062.

[0127] Advantageously, the microwave extraction device may also include conventional means for performing partial or total vacuum extraction. Preferably, the partial vacuum may advantageously be between 50 mb and 300 mb.

[0128] In the same extraction cycle, the microwave extraction step (b1) may include one or more microwave treatment sequences, identical or different, particularly in terms of their power, temperature and / or duration.

[0129] According to a particular embodiment, the device used for the microwave extraction step is the one marketed under the names E200 to E2000, ECT200 to ECT1000 or ECS150 to ECS1000 by IDCO, or the one marketed under the name EMU06 by SAIREM. In particular, a device for the microwave extraction step is the E200 marketed by IDCO.

[0130] A person skilled in the art will be able to adjust / adapt the device if necessary, especially when microwave extraction is carried out in native water, i.e. without the addition of extraction solvent.

[0131] Step (b2) - second maceration extraction step

[0132] According to a first embodiment, the maceration extraction step can be carried out without the addition of an extraction solvent, using only the first aqueous extract, which is particularly advantageous. Thus, the extraction solvent considered for this maceration extraction step is the first aqueous extract obtained at the end of step (b1).

[0133] According to a second embodiment, the extraction step (b2) is carried out in the presence of at least extraction solvent.

[0134] When step (b1) and step (b2) use at least one extraction solvent, the latter may be the same or different.

[0135] Naturally, a person skilled in the art will ensure that the extraction solvent(s) and / or their quantity are chosen in step (b1) and / or step (b2) in such a way that the advantageous properties of a process according to the invention, and in particular of the extract according to the invention, are not or are not substantially altered by the envisaged addition. Likewise, a person skilled in the art will ensure that the nature and / or quantity of the extraction solvent(s) is chosen according to the molecule(s) or active agent(s) that are to be extracted.

[0136] For illustrative purposes, an extraction solvent usable in step (b1) and / or in step (b2) may be chosen from water, at least one alcohol, a hydro-alcoholic solvent, and mixtures thereof, and particularly chosen from glycerin, water, ethanol, isopropanol, propanediol, butylene glycol, pentylene glycol, propylene glycol, and mixtures thereof.

[0137] Preferably, step (b2) of extraction by maceration can be carried out:

[0138] - for a period of between 1 minute and 300 minutes (or 5 hours), in particular between 5 minutes and 300 minutes (or 5 hours), preferably between 10 minutes and 240 minutes (or 4 hours), better between 10 minutes and 180 minutes (or 3 hours), especially between 1 hour and 120 minutes (or 2 hours), or even between 20 minutes and 40 minutes; and / or

[0139] - carried out hot, preferably at a temperature between 50°C and 90°C, preferably between 60°C and 80°C, and especially between 65°C and 75°C.

[0140] In the same extraction cycle, the maceration extraction step (b2) may include one or more maceration treatment sequences, identical or different, particularly with regard to temperature, nature and / or quantity of extraction solvent(s), and / or duration.

[0141] According to a first variant, the step (b2) of extraction by maceration is carried out cold.

[0142] When step (b) includes step (b1) and step (b2), step (b2) of maceration extraction is advantageously carried out with, and within, the same device as that used for step (b1) of microwave extraction.

[0143] Preferably, the maceration extraction device may also include at least:

[0144] - a solution for mixing the plant to be treated; and / or

[0145] - a rotating tank forming a cavity preferably made of a refractory metal with low resistivity, driven in rotation by means allowing its speed to be varied in order to optimize the mixing of the plant being treated. Such an embodiment is described in particular in EP2530059.

[0146] Additional Step(s) According to a particular embodiment, the extraction process according to the invention may further comprise, during step (b1), between steps (b1) and (b2), and / or after step (b2), at least one cooling step (g). This cooling step may be carried out such that the first aqueous extract and / or the extract of interest is discharged by tipping into a cooler, for example, with air, the airflow of which is ensured, for example, by a fan. One purpose of this cooler is to accelerate the cooling of the material. Another purpose of this cooler is to recover the heat emitted by the processed material after its exit from the chamber, during the cooling step. The cooling of the processed material may, for example, be carried out by circulating an airflow over it, for example, by means of a fan or any other equivalent means.

[0147] In one particular embodiment, the extraction process includes at least one cooling step (g) carried out simultaneously with step (b1), and possibly also with step (b2), in the tank (or receptacle) containing the plant. In this embodiment, the tank of the device is advantageously equipped with a double jacket, with the cooling occurring in the outer jacket. This embodiment is advantageous because it allows, in particular, the condensation of volatile matter extracted during step (b1), and thus optimizes the composition and / or the extraction yield of the first aqueous extract, and therefore of the extract of interest.

[0148] According to another particular embodiment, an extraction process according to the invention may further comprise, before step (b), between steps (b1) and (b2), and / or after step (b2), and preferably after step (b2), at least one additional pressing step (e). This pressing step, carried out on the plant, the first aqueous extract, and / or the extract of interest, and preferably on the extract of interest, may be performed using any suitable device, and in particular using a press. This pressing step ensures solid / liquid separation.

[0149] According to another particular embodiment, an extraction process according to the invention may further comprise, after step (b), and in particular after step (b2), at least one additional filtration step (c). This filtration, carried out on the extract of interest, may be performed using any suitable device, in particular a sieve or a filter. Advantageously, the filtration allows the "juice" (or "filtrate") of the extract of interest to be separated from any fibers and other solid plant elements larger than the pores of the filter device. Preferably, this filtration operation removes from the extract of interest any element larger than 0.2 microns (µm), which corresponds to the debacterial threshold. Preferably, the defiltration step (c) is carried out using a plate filter, which allows filtration to be reduced, preferably down to 0.2 microns, by a cascade of steps.

[0150] Advantageously, a production process according to the invention comprises, after step (b), and preferably after step (b2), at least one pressing step (e) and / or at least one filtration step (c). Preferably, when the production process according to the invention comprises, after step (b), and preferably after step (b2), at least one pressing step (e) and at least one filtration step (c), the pressing step (e) is carried out before the filtration step (c), or vice versa.

[0151] According to yet another particular embodiment, an extraction process according to the invention may further comprise, after step (b), and in particular after step (b2), at least one additional step (d) of mixing the extract, optionally previously pressed and / or filtered, with at least one solvent, in particular a preservation solvent.

[0152] Preferably, a preservative solvent can be chosen from glycerin, propanediol (or propylene glycol), butylene glycol, pentylene glycol, and mixtures thereof, and preferably glycerin. This step (f) is advantageous in that it prevents bacterial growth, thus improving the preservation of the product obtained from the extraction process according to the invention.

[0153] Preferably, the extract of interest is mixed with at least one preservation solvent in an "extract of interest / preservation phase" ratio of between 10 / 90 and 95 / 5, preferably between 10 / 90 and 60 / 40, better between 20 / 80 and 50 / 50, and in particular between 20 / 80 and 40 / 60.

[0154] Preferably, a process according to the invention may further comprise at least one step (i) of decolorizing the extract. The decolorization may be carried out by adsorption of any pigments present in the extract or filtrate onto activated carbon under agitation for a period of between 1 and 6 hours, preferably between 2 and 4 hours, at room temperature. Preferably, the activated carbon represents 30% by mass relative to the dry matter of the extract. This decolorization step may be followed by a filtration step of the resulting decolorized filtrate, for example, by passing it through a 4 µm sieve or filtration membrane.

[0155] Preferably, when step (b) is carried out in the presence of at least one aqueous, alcoholic, or hydroalcoholic solvent, a process according to the invention may further comprise, between steps (b) and (d), or even between steps (c) and (d), at least one solvent removal step (h), preferably by evaporation. Preferably, this step (h) is carried out by partial or total evaporation under reduced pressure, for example, using a rotary evaporator or a scraped-film evaporator. This step (h) is particularly advantageous in that it allows the extract according to the invention to be concentrated while ensuring the absence of hazardous and / or polluting solvents, even in trace amounts.

[0156] Advantageously, an extraction process according to the invention:

[0157] - does not include, before step (b), a step of adding solvent(s), in particular hydrophilic (or aqueous) and / or lipophilic and / or amphiphilic solvent(s);

[0158] - does not include an oil addition step;

[0159] - does not include a step of contact with supercritical CO2;

[0160] - does not include a separation step between an oil phase and an aqueous phase; and / or

[0161] - does not include a heating step under an atmosphere devoid of, or essentially devoid of, C>2.

[0162] Extract

[0163] For the purposes of this invention, the extract comprises at least one active agent. For the purposes of this invention, "active agent" means a compound having a beneficial physiological effect on the element on which it acts. Its purpose may be, for example, to protect, maintain, treat, cure, perfume, flavor, or color. The extract of interest is advantageously a cosmetic, dermo-pharmaceutical, pharmaceutical, or food-grade agent.

[0164] Preferably, an active agent is chosen from polyphenols, proteins, amino acids, peptides, sugars, minerals, vitamins, carotenoids, enzymes, and mixtures thereof, and especially.

[0165] Preferably, an active agent is a polyphenol (or "plant tannin," or "biophenol"). Polyphenols are a family of complex molecules that plants naturally produce to defend themselves against various stresses. They are characterized by the presence of at least two phenols linked together in more or less complex structures, generally of high molecular weight.

[0166] A polyphenol can be chosen from among phenolic acids (catechuic acid, gallic acid, protocatechuic acid), flavones, chlorogenic acid, quinones, rosmarinic acid and their mixtures, and especially rosmarinic acid.

[0167] A polyphenol can be chosen from among the simple phenols, in particular phenolic acids and coumarins; naphthoquinones; stilbenoids (two C6 rings linked by two carbon atoms); flavonoids, in particular isoflavonoids and anthocyanins; polymerized forms, in particular lignans, lignins, condensed tannins; and mixtures thereof. Also, polyphenols are commonly subdivided into flavonoids or non-flavonoids (resveratrol, phenolic acids, lignans).

[0168] A flavonoid-type polyphenol can be selected from a flavone (e.g., quercetin), a flavonol (e.g., catechin), an anthocyanidin, an isoflavone, a flavonone, a catechin, and mixtures thereof.

[0169] A non-flavonoid type polyphenol can be chosen from among stilbenes (e.g. resveratrol), phenolic acids, lignans, and mixtures thereof.

[0170] Preferably, the active agent is rosmarinic acid.

[0171] As stated above, the extract according to the invention is an aqueous, hydro-alcoholic or alcoholic extract of at least one plant or at least part of at least one plant of the genus Perilla, comprising at least 2%, preferably at least 3%, in particular at least 4%, better at least 5%, and most particularly at least 6%, by weight of total polyphenols, and most particularly at least 0.2%, preferably at least 0.3%, and in particular at least 0.4%, by weight of rosmarinic acid, relative to the total dry weight of the extract.

[0172] Composition

[0173] Preferably, the extract according to the invention (or "extract of interest") can be used in the fields of cosmetics, nutraceuticals, food, or pharmaceuticals.

[0174] Thus, an extract of interest can be implemented in a composition, in particular a cosmetic, pharmaceutical, nutraceutical or food composition, optionally in association with at least one physiologically acceptable medium.

[0175] Preferably, the composition according to the invention may comprise between 0.01% and 20%, preferably between 0.1% and 10%, better between 0.5% and 5%, and most particularly between 1% and 3%, by weight of extract(s), relative to the total weight of the composition.

[0176] A composition according to the invention can in particular be adapted for application by oral route or by topical route, preferably topical.

[0177] Preferably, a composition according to the invention is a cosmetic composition, in particular dedicated to the care and / or makeup of a keratinous material, and in particular of the skin and / or hair.

[0178] In particular, the composition of the invention may be in the form of an anti-aging skincare composition, especially for the face.

[0179] According to another embodiment, a composition of the invention may be in the form of a makeup composition, in particular in the form of a foundation, a tinted base or a tinted cream. A composition according to the invention may further comprise at least one physiologically acceptable medium (or solvent), chosen in particular according to the different ingredients, the form of administration and / or the cosmetic, pharmaceutical, nutraceutical or food purpose.

[0180] In the context of the invention, and unless otherwise stated, "physiologically acceptable medium" means a medium suitable for the application considered, in particular for cosmetic applications, and suitable in particular for the application of a composition according to the invention on a keratinous material, in particular the skin and / or hair, and more particularly the skin.

[0181] The physiologically acceptable medium is generally adapted to the nature of the support on which the composition is to be applied, as well as to the appearance in which the composition is to be packaged.

[0182] When the application in question is a cosmetic application, the physiologically acceptable environment is particularly compatible with the skin, mucous membranes and hair.

[0183] Examples include water (preferably demineralized water or floral waters), optionally in combination with at least one preservative and / or alcohol.

[0184] A composition comprising at least one extract according to the invention may be in any form. Such a composition may be, for example, a cream, an emulsion (direct, inverse or multiple), a lotion, a serum, a gel and an oil for the skin (hands, face, feet, etc.), a foundation (liquid, paste), a preparation for baths and showers (salts, foams, oils, gels, etc.), a hair care product (hair dyes and bleaches), a cleaning product (lotions, powders, shampoos), a hair conditioning product (lotions, creams, oils), a styling product (lotions, hairsprays, brilliantines), a shaving product (soaps, foams, lotions, etc.), a product intended to be applied to the lips, a sunscreen, a sunless tanning product, a skin whitening product, an anti-wrinkle product.In particular, a cosmetic composition can be an anti-aging serum, a youth serum, a hydrating serum or a perfumed water.

[0185] A food composition comprising at least one extract according to the invention may be, for example, a yogurt or a food supplement, in particular a liquid one.

[0186] Additional component(s)

[0187] An extract, or even a composition comprising at least said extract, according to the invention, may further comprise at least one additional compound different from the aforementioned extract and physiologically acceptable medium. An extract, or even a composition, according to the invention, may further comprise at least powders, organic or inorganic; hydrophilic texturizing agents (or hydrophilic gelling agents), in particular as described in FR3041251; film-forming agents; coloring agents, in particular selected from water-soluble or insoluble, fat-soluble or insoluble, organic or inorganic coloring agents, optical effect materials, liquid crystals, and mixtures thereof; fillers, in particular pigments and / or pearlescent pigments, in particular as described in FR3067930 and / or blurring effect fillers / "soft-focus" fillers, in particular as described in WO2019053236;Bulking agents such as lauroyl-lysine, starch, boron nitride and / or silica; emulsifying and / or non-emulsifying silicone elastomers, particularly those described in EP2353577; texturizing agents; surfactants; glycerin; preservatives; salts; humectants; stabilizers; pH stabilizing agents, in particular a pH buffer (e.g., HEPES, PBS); chelating agents; oils; emollients; retarding agents; etc.; or any common cosmetic additive; and mixtures thereof.

[0188] An extract, or even a composition, according to the invention, may further comprise at least one biological / cosmetic active ingredient (or active agent) different from the extract of interest. The active agent is advantageously a cosmetic, dermo-pharmaceutical, pharmaceutical, or food-grade agent.

[0189] A biological / cosmetic active ingredient can be chosen from among moisturizing agents, healing agents, depigmenting agents, UV filters, desquamating agents, antioxidant agents, active ingredients stimulating the synthesis of dermal and / or epidermal macromolecular molecules, dermo-contracting agents, anti-perspirant agents, soothing agents, anti-wrinkle agents, anti-irritant agents, tightening / smoothing agents, anti-cellulite agents, firming agents, sheathing agents, softening agents, emollient agents, slimming agents, protective / anti-radical agents, draining agents, anti-inflammatory agents, whitening agents, self-tanning agents, exfoliating agents, agents stimulating cell renewal or stimulating cutaneous microcirculation, absorbing agents, anti-dandruff agents and / or anti-aging agents, and their mixtures.

[0190] When the active agent is a pharmaceutical agent, it is advantageously chosen from among anticoagulants, antithrombogenics, antimitotic agents, antiproliferative agents, antiadhesion, antimigration agents, cell adhesion promoters, growth factors, antiparasitic molecules, anti-inflammatories, angiogenics, angiogenesis inhibitors, vitamins, hormones, proteins, antifungals, antimicrobial molecules, antiseptics, antibiotics, and mixtures thereof.

[0191] The food-grade agents are advantageously vegetable or fruit purees such as mango puree, pear puree, coconut puree, cream of onions, leeks, carrots, or other preparations that can combine several fruits or vegetables. Alternatively, they are oils such as edible oils, like olive oil, soybean oil, grapeseed oil, sunflower oil, or any other oil extracted from plants, as well as food-grade active ingredients such as probiotics, yeasts, vitamins, minerals, oleoactives, and mixtures thereof.

[0192] An extract, or even a composition, according to the invention, may further comprise excipients, such as thickeners, or rheology modifiers. These thickeners are, for example, polymers, cross-polymers, microgels, gums or proteins, including polysaccharides, celluloses, polysaccharides, silicone-based polymers and copolymers, colloidal particles (silica, clays, latex, etc.), and mixtures thereof.

[0193] The additional compound(s), in particular the active ingredient(s), may be hydrophilic or lipophilic.

[0194] Naturally, a person skilled in the art will take care to choose any additional compound(s) and / or their quantity in such a way that the advantageous properties of a composition according to the invention, and in particular of the extract according to the invention, are not or substantially not altered by the proposed addition. These adjustments fall within the general knowledge of a person skilled in the art.

[0195] Preferably, a composition according to the invention is free of surfactant.

[0196] Uses

[0197] According to another aspect, the invention relates to the non-therapeutic use, particularly cosmetic, of an extract according to the invention, to prevent and / or combat skin aging, in particular photo-induced skin aging, improve cell longevity, restore the skin barrier function and / or improve skin hydration.

[0198] For the purposes of this invention, "cosmetic" means a non-pharmaceutical, non-therapeutic use which is not intended for therapeutic use, nor to prevent or treat symptoms related to a therapeutic skin condition.

[0199] Indeed, advantageously, the Applicant found that an extract according to the invention possessed several activities of interest with respect to preventive or repairing physiological mechanisms related to the skin barrier function, hydration and / or skin cell longevity.

[0200] As can be seen from the examples below, the Applicant notably found, advantageously and unexpectedly, that an extract according to the invention made it possible to stimulate biological pathways related to the skin barrier function and in particular markers of skin aging, and especially of skin photo-aging, such as ceramides, Aquaporin 3 (AQP3), Filaggrin and Claudin 1, as well as a significant reduction in lipid peroxidation.

[0201] Throughout the description, including the claims, the expression "comprising one" shall be understood as synonymous with "comprising at least one", unless otherwise specified.

[0202] The expressions "between ... and ...", "from ... to ..." and "ranging from ... to ..." should be understood inclusive of boundaries, unless otherwise specified.

[0203] The following examples illustrate the present invention without limiting its scope. Unless otherwise stated, the quantities of ingredients shown in the examples are expressed as a percentage by weight of the total weight of the composition. FIGURES

[0204] Figure 1 shows the extraction results for total polyphenols (TPC) and rosmarinic acid (RA) by weight. In this figure 1, the extraction carried out at room temperature is designated by the expression "non-optimized extract" and the extraction carried out at 70°C is designated by the expression "optimized extract".

[0205] Figure 2 illustrates a bar chart representing MDA-TBARS levels normalized to cell viability after exposure to UV light (4 J / cm²). 2 UVA, 374 mJ / cm 2UVB) compared to the control not exposed to UV light. The asterisk (*) above the bar indicates a significant difference compared to the unexposed control. The hash symbol (#) indicates a significant difference compared to the control + UV (unpaired Student's t-test).

[0206] Figure 3 illustrates a bar chart representing total CER levels relative to control + UV, after 72 hours of topical treatment of RHE tissues with Red Shiso extract according to Example 2 and subsequent exposure to UV light (4 J / cm²). v ) UVA, 374 mJ / cm A (2) UVB). The asterisks (*) above the bars indicate statistical significance compared to the control + UV (Dunnett's post hoc multiple comparison test).

[0207] Figure 4 illustrates a bar chart representing AQP3 levels after 24 hours of topical treatment of RHE tissues with Red Shiso extract according to example 2, followed by subsequent exposure to UV light (4 J / cm²). 2 UVA, 374 mJ / cm 2 UVB). Asterisks (*) above the bars indicate a statistically significant result compared to the control + UV (Dunnett post hoc multiple comparison test).

[0208] Figures 5A and 5B show a significant decrease in Filaggrin expression of 24.5% after UV exposure. A significant increase (+139%, p=0.045) in Filaggrin expression is observed after systemic application of the Red Shiso extract according to Example 2, at a rate of 0.3 times higher than after UV exposure.

[0209] Figures 6A and 6B show a significant increase (2.4x) in Claudin 1 expression after systemic application of the Red Shiso extract according to Example 2 at 0.3x compared to UV exposure. EXAMPLES

[0210] Example 1 - culture of Perilla frutescens (i.e. step (a) of a process according to the invention)

[0211] This example 1 relates to step (a) of a process for obtaining at least one extract according to the invention.

[0212] Before being placed in a tray (or Trayplant), i.e. pots or cells grouped on a tray, the seeds undergo a stratification stage at 4°C for 3 days in order to lift the dormancy of the seeds and ensure a satisfactory level of germination.

[0213] As a substrate usable for the cultivation of Perilla frutescens, one can use a Coco substrate, a Peat substrate or a Hemp substrate, preferably a Coco substrate which is the one which shows the most interesting g / tray yield ("g / tray yield" means the quantity of plant harvested, in grams, by each plant which was initially grown in a tray).

[0214] The seeds of the plant species Perilla frutescens on Coco substrate are then cultivated in hydroponic culture, and therefore under controlled conditions in terms of water supply, humidity, temperature, and light supply.

[0215] Tables 1 and 2 below present data on the impact of light input on the production of total polyphenols (TPC) and rosmarinic acid in particular. These data were obtained from fresh material harvested on day 21 after culturing.

[0216] The biomass extraction protocol for the analysis of total polyphenol (TPC) and rosmarinic acid content is as follows:

[0217] After 21 days of cultivation, the Perilla frutescens plants are harvested and immediately frozen.

[0218] Frozen fresh Perilla frutescens plants are then ground and mixed with a 60% MeOH solution with a solid / liquid ratio of 1 / 25;

[0219] Total polyphenols (TPCs) and rosmarinic acid are extracted by ultrasound for 30 minutes; and

[0220] TPC levels were determined using the Folin-Ciocalteur method (1999, Singleton et al.), and rosmarinic acid levels were determined by UPLC-UV. Impact of photoperiod on production

[0221]

[0222] * 18 / 6 means that, over 24 hours, the plants in cultivation are illuminated for 18 hours and are in darkness for 6 hours.

[0223] The results in Table 1 above show a significant impact of photoperiod on crop yield, but also on the synthesis of total polyphenols (TPC) and in particular rosmarinic acid.

[0224]

[0225] *Tests carried out from test 2 of table 1, namely with a photoperiod of 24 / 0.

[0226] The results in Table 2 above show a significant impact of light intensity on crop yield, but also on the synthesis of total polyphenols (TPC) and in particular rosmarinic acid.

[0227] Example 1 shows that growing conditions, and in particular the supply of light, have a significant impact on crop yield, but also on the synthesis of total polyphenols (TPC) and in particular rosmarinic acid.

[0228] Example 2 - Extraction (i.e. step (b) of a process according to the invention) Example 2 is carried out using plants according to test 2.2 of example 1, harvested at 21 days of culture, and frozen in a refrigerator at -20°C for a time necessary and sufficient for proper freezing of the material.

[0229] The aqueous extraction process implemented in this example 2 is based on an extraction cycle comprising: (b1) a first microwave-assisted extraction step carried out with an IDCO 200 machine; and

[0230] (b2) a second maceration extraction step applied to the first aqueous extract obtained in step (b1) and still in the container used in step (b1).

[0231] In step (b1), 600 grams of plant material are mixed with a water solvent in a liquid / solid ratio of 1 / 1.

[0232] Step (b1) is carried out at room temperature (TA) or hot (70°C), with stirring, for 15 minutes, at a power of 600 W.

[0233] Next, step (b2) of maceration extraction consisted of leaving the mixture obtained at the end of step (b1) in the same container under agitation at room temperature (TA) or at 70°C, for a further 15 minutes.

[0234] Finally, a liquid / solid separation of the mixture obtained at the end of step (b2) is carried out by filtration under press and then plate filter, the "juice" obtained is then mixed with a preservation solvent (i.e. glycerin) in a "juice / glycerin" ratio of 20 / 80.

[0235] We then carry out the determination of total polyphenols (TPC) and rosmarinic acid (RA) according to the methods described previously.

[0236] Figure 1 shows the extraction results for total polyphenols (TPC) and rosmarinic acid (RA) by weight. In this figure 1, the extraction carried out at room temperature is designated by the expression "non-optimized extract" and the extraction carried out at 70°C is designated by the expression "optimized extract".

[0237] The test plan described in Example 2 therefore significantly improved the total polyphenol concentrations and the rosmarinic acid concentration. Example 2 shows more specifically that the extraction conditions, and in particular the temperature parameter, have a significant impact on the concentration of total polyphenols (TPC) in the final extract, and especially on the concentration of rosmarinic acid.

[0238] Example 3 - Objectification and characterization of the cosmetic effects of the extract according to example 2

[0239] Example 3 involves studying the effects of the Red Shiso extract obtained in example 2 on 2 3D models of human skin, namely the reconstituted human epidermis (RHE) and expiants (ex vivo) that have undergone UV treatments, in order to observe its impact on lipid peroxidation, ceramide levels, the expression levels of aquaporin 3, filaggrin and Claudin 1, all known to be commonly used biomarkers for assessing skin aging, particularly photo-induced skin aging, skin barrier function and skin hydration.

[0240] Indeed, as previously mentioned, ultraviolet (UV) radiation is considered one of the main causes of skin aging, particularly due to its effects on oxidative stress. Lipids, vital components of skin structure and function, are especially sensitive to the harmful effects of oxidative stress.

[0241] Lipid peroxidation, a cascade of oxidative reactions initiated by ROS, leads to lipid degradation, resulting in structural alterations and cellular dysfunction, and thus plays a central role in skin aging. At the molecular level, lipid peroxidation generates numerous byproducts, including malondialdehyde (MDA) and thiobarbituric acid reactive substances (TBARS). MDA and TBARS are commonly used markers to assess the extent of lipid peroxidation (Piao et al., 2018; Aguilar Diaz De Leon and Borges, 2020).

[0242] • In addition to direct oxidative damage, UV radiation is detrimental to the regulation of several molecules linked to the epidermal barrier function, and in particular ceramides (CERs), which help maintain the barrier function through their hydrophobic properties. A link between the alteration of barrier function occurring during photoaging and a dysfunction in the levels or proportion of CERs in the epidermis is known (Takagi et al., 2004; Yoon et al., 2019; Ahn et al., 2022; Barresi et al., 2022).

[0243] Aquaporin 3 (AQP3) is a member of the aquaporin family, composed of 13 protein members that allow the selective penetration of water into cell membranes. AQP3 transports water, glycerol, and hydrogen peroxide and is expressed throughout the body. In the skin, AQP3 is expressed in keratinocytes and plays an important role in skin hydration and maintaining the skin barrier (Bollag et al., 2020). The loss of AQP3 in skin cells exposed to UV radiation is an underlying cause of the dehydration observed during photoaging (Tang et al., 2019; Bae et al., 2019; Cao et al., 2008).

[0244] • Another molecule altered by UV radiation is filaggrin, a structural protein in the stratum corneum of the epidermis where it plays a dual role. On the one hand, it enables the aggregation of keratin filaments in keratinocytes, leading to the formation of corneocytes; on the other hand, it provides natural hydration factors (Kim and Lim, 2021). Filaggrin therefore plays a crucial role in the skin barrier. UV radiation induces a decrease in filaggrin expression and causes premature keratinization of corneocytes, resulting in a weakened and dehydrated skin barrier (Scott, 1986; Sandilands et al., 2009; Thyssen et al., 2014).

[0245] Finally, claudin-1 is a transmembrane protein expressed in all layers of the epidermis and involved in cell adhesion (Tokumasu et al., 2017). It is the main component of tight junctions, structures essential for the skin's barrier function. In the presence of UV radiation, claudin-1 is fragmented into a discontinuous membrane motif, leading to a weakening of the barrier function (Takuo et al., 2011).

[0246] 3.1) RHE model procedure:

[0247] The reconstructed human epidermis (RHE) model allows us to evaluate the effects of the Red Shiso extract obtained in example 2 on lipid peroxidation, ceramides and aquaporin 3.

[0248] The experiment was conducted on an RHE SkinEthic™ RHE / S / 17 model subjected to topical application of Red Shiso extract according to Example 2 at a concentration of 0.3% for 72 hours. Simultaneously, the skin was exposed to ultraviolet (UV) radiation (total dose: UVA 4 J / cm²). A (2); UVB 374 mJ / cm A (2)). After treatment, the percentage of lipid peroxidation was measured as an index of antioxidant activity. Furthermore, the tissues were fixed, cryosectioned, and then hybridized with anti-CERAMIDES or anti-AQUAPORIN 3 antibodies, followed by a secondary conjugated antibody, AlexaFluor488.

[0249] 3.1.1) Results - effects on antioxidant activity

[0250] Figure 2 illustrates a bar chart representing MDA-TBARS levels normalized to cell viability after exposure to UV light (4 J / cm²). A (2) UVA, 374 mJ / cm A(2) UVB) compared to the control not exposed to UV light. The asterisk (*) above the bar indicates a significant difference compared to the unexposed control. The hash symbol (#) indicates a significant difference compared to the control + UV (unpaired Student's t-test).

[0251] Figure 2 shows that UV irradiation significantly increased MDA-TBAR levels in RHE tissues by 4.7 times compared to the untreated, unirradiated control.

[0252] When tissues were treated with Red Shiso extract at a concentration of 0.3%, the level of MDA-TBARs decreased significantly by -18.4% compared to the control + UV. 3.1.2) Results - effects on ceramides (CER)

[0253] Figure 3 illustrates a bar chart representing total CER levels relative to control + UV, after 72 hours of topical treatment of RHE tissues with Red Shiso extract according to Example 2 and subsequent exposure to UV light (4 J / cm²). A (2) UVA, 374 mJ / cm A (2) UVB). The asterisks (*) above the bars indicate statistical significance compared to the control + UV (Dunnett's post hoc multiple comparison test).

[0254] These results show that UV irradiation applied to RHE alters the level of ceramides, which decrease non-significantly by 7.1% compared to the untreated and unirradiated control.

[0255] Treating UV-exposed tissues with 0.3% Red Shiso extract according to example 2 led to a statistically significant increase in CER levels in these tissues (+44.6%) compared to the control + UV.

[0256] 3.1.3) Results - effects on aquaporin 3

[0257] Figure 4 illustrates a bar chart representing AQP3 levels after 24 hours of topical treatment of RHE tissues with Red Shiso extract according to example 2, followed by subsequent exposure to UV light (4 J / cm²). A (2) UVA, 374 mJ / cm A (2) UVB). Asterisks (*) above the bars indicate a statistically significant result compared to the control + UV (Dunnett post hoc multiple comparison test).

[0258] UV exposure resulted in a statistically significant decrease of 13.2% compared to the non-irradiated control. Treatment of UV-exposed tissues with Red Shiso extract according to Example 2 resulted in a statistically significant increase in AQP3 levels at the dose of 0.3% (+17.6%) compared to the control + UV.

[0259] 3.2) Ex vivo procedure

[0260] The ex vivo model allows us to evaluate the effects of the Red Shiso extract according to example 2 on Filaggrin and Claudin 1.

[0261] Samples of healthy human skin from 6 subjects of Caucasian origin were cut into 1 cm skin fragments 2and placed in inserts positioned on culture wells, with the epidermis in contact with open air. The entire system is maintained in survival mode at 37°C in an air / CO2 atmosphere (95% / 5%). Culture medium (Modified Dulbecco Minimum Essential Medium; Gibco BRL, USA), enriched with antibiotics (100 pg / ml penicillin, 100 pg / ml streptomycin; Gibco BRL, USA), hormones (SIGMA, France), bovine pituitary extract (Gibco BRL, USA), and SVF (DAP, France), is placed in the bottom of the wells. Passage occurs by slow diffusion between the two compartments to the dermis via the porous membrane (3 µm). A skin oxidation model was created by a session of UVA (8 J / cm²) and UVB (0.5 J / cm²) radiation at 0°C. Immediately after the UV session, the Red Shiso extract according to example 2 was added daily at 2 concentrations in the culture medium for 3 days (J0, J1 and J2).On day 3, the skin fragments were fixed in formalin and embedded in paraffin for immunohistochemical analysis of Filaggrin and Claudine 1.

[0262] Filaggrin detection (rabbit polyclonal antibody, source: St John's Laboratory) was performed using a 2-layer immunoperoxidase technique (Impress kit, Vector Laboratories) and revealed in red by 3-amino-9-ethylcarbazole (AEC).

[0263] Claudin 1 (rabbit polyclonal antibody, clone PO4679, Eu), expressed in the granulocytic layer and the upper 1 / 3 of the epithelium, was detected using an indirect 3-layer immunoperoxidase technique (Polink-2 HRP kit, Origene, Diagomics) and revealed in red by AEC.

[0264] 3.2.1) Results - effects on Filaggrin

[0265] Figures 5A and 5B show a significant decrease in Filaggrin expression of 24.5% after UV exposure (Figure 5A-5B(b) vs. Figure 5A-5B(a)). A significant increase (+139%, p=0.045) in Filaggrin expression is observed after systemic application of the Red Shiso extract according to Example 2 at 0.3 compared to UV exposure (Figures 5A-5B(c) vs. Figures 5A-5B(b)).

[0266] 3.2.2) Results - effects on Claudine 1

[0267] Figures 6A and 6B show a significant increase (x2.4) in Claudin 1 expression after systemic application of Red Shiso extract according to example 2 at 0.3 compared to UV aggression (figures 6A-6B (c) vs. figure 6A-6B (b)).

[0268] CONCLUSIONS:

[0269] The results above show that an extract of Red Shiso according to the invention induces:

[0270] 1- a significant reduction in lipid peroxidation;

[0271] 2- an increase in ceramides;

[0272] 3- an increase in the expression of the aquaporin 3 protein; 4- an increase in the expression of the filaggrin protein; and 5- an increase in the expression of the Claudin 1 protein. Therefore, these results demonstrate that a Red Shiso extract according to the invention makes it possible to prevent and / or effectively combat skin aging, in particular photo-induced skin aging, restore the skin barrier function and improve skin hydration.

Claims

34 DEMANDS 1. Aqueous, hydro-alcoholic or alcoholic extract of at least one plant or at least part of at least one plant of the genus Perilla, said extract comprising at least 2%, preferably at least 3%, in particular at least 4%, better at least 5%, and most particularly at least 6%, by weight of total polyphenols, and most particularly at least 0.2%, preferably at least 0.3%, and in particular at least 0.4%, by weight of rosmarinic acid, relative to the total dry weight of the extract.

2. Extract according to claim 1, wherein the extract comprises less than 0.1% by weight, in particular less than 0.01% by weight, relative to the total weight of said extract, of organic solvent, in particular selected from the group consisting of methanol, ethanol, acetone, ethyl acetate, chloroform, dichloromethane and mixtures thereof.

3. Extract according to claim 1 or 2, wherein the extract is free from organic solvent, and in particular free from methanol, ethanol, acetone, ethyl acetate, chloroform, and dichloromethane.

4. Extract according to any one of the preceding claims, wherein the extract is an aqueous extract.

5. Extract according to any one of the preceding claims, wherein the plant is of the species Perilla frutescens.

6. Extract according to any one of the preceding claims, wherein the extract is obtained from at least one plant or at least part of at least one plant having a growth stage between 5 and 30 days, preferably between 7 and 25 days, and better between 10 and 22 days.

7. Extract according to any one of the preceding claims, wherein the extract is obtained from the roots of the plant, the aerial parts of the plant, and mixtures thereof, and preferably from the aerial parts of the plant, and even more preferably from the leaves, stems, flowers and flowering tops, buds, and mixtures thereof.35 8. Extract according to any one of the preceding claims, wherein the extract is obtained from at least one plant or from at least a part of at least one plant having been cultivated: - under irradiation at a light intensity between 100 and 600 pmol / cm² 2 / s, preferably between 150 and 500 pmol / cm 2 / s, and better between 200 and 400 pmol / cm 2 / s ; - with a photoperiod in which exposure to light is between 16 hours and 24 hours, preferably between 18 hours and 24 hours, better between 20 hours and 24 hours, and particularly between 22 hours and 24 hours; and / or - under irradiation at a light intensity having a spectrum comprising between 70% and 40% of the flux in a wavelength between 400 nm and 500 nm, between 30% and 0% between 500 nm and 600 nm, between 50% and 10% between 600 nm and 700 nm and between 20% and 0% between 700 nm and 800 nm.

9. A method for obtaining at least one extract according to any one of the preceding claims, characterized in that it comprises at least the following steps according to which: (a) at least one plant or at least part of at least one plant of the genus Petilla is available, optionally in crushed and / or frozen form; (b) the plant or plant part from step (a) is subjected to at least one extraction cycle comprising: (b1) at least one microwave extraction step, optionally with stirring; and / or (b2) at least one maceration extraction step, optionally under agitation; preferably step (b1) and step (b2), when both present, being carried out within the same extraction device; (c) Optionally, the extract obtained in step (b) is purified by filtration, ultrafiltration and / or nanofiltration; and (d) Optionally, the extract obtained in step (b), or in step (c), is redispersed in a preservation solvent.

10. A process according to the preceding claim, wherein said process does not include a concentration step and / or a purification step of the extract obtained in step (b), (c) or (d).

11. A process according to claim 9 or 10, wherein the plant of step (a) has been cultivated under controlled conditions, preferably in a cultivation unit ensuring control of the growing conditions, in particular ensuring controlled regulation of water, humidity, temperature, and air quality and light supply, and in particular wherein the plant of step (a) has been obtained by hydroponic or aquaponic culture.

12. A method according to any one of claims 9 to 11, wherein step (b), in particular step (b1), is carried out using at least one plant or at least a part of at least one plant: - fresh, and preferably a frozen fresh plant; and / or - previously dried, freeze-dried, frozen and / or ground.

13. A process according to any one of claims 9 to 12, wherein step (b1) and / or step (b2) is / are carried out without the addition of extraction solvent or is / are carried out in the presence of at least one extraction solvent, identical or different, and preferably selected from water, at least one alcohol, a hydro-alcoholic solvent, and mixtures thereof, and particularly selected from glycerin, water, ethanol, isopropanol, propanediol, butylene glycol, and mixtures thereof.

14. A method according to any one of claims 9 to 13, wherein the microwave extraction step (b1) is carried out: - at a power between 600 W and 6000 W, preferably between 700 W and 5000 W, better between 800 W and 4000 W, and especially between 1000 W and 2000 W, or even between 2000 W and 6000 W; - for a period of between 1 minute and 180 minutes, preferably between 2 minutes and 120 minutes, better between 5 minutes and 100 minutes, in particular between 10 minutes and 60 minutes, or even between 15 minutes and 30 minutes; and / or - at a temperature between 30°C and 120°C, preferably between 40°C and 100°C, and better between 50°C and 90°C, preferably between 60°C and 80°C, and especially between 65°C and 75°C.

15. A method according to any one of claims 9 to 14, wherein the maceration extraction step (b2) is carried out: - for a period of between 1 minute and 300 minutes, in particular between 5 minutes and 300 minutes, preferably between 10 minutes and 240 minutes, better between 10 minutes and 180 minutes, especially between 1 hour and 120 minutes, or even between 20 minutes and 40 minutes; and / or - at a temperature between 50°C and 90°C, preferably between 60°C and 80°C, and particularly between 65°C and 75°C.

16. A process according to any one of claims 9 to 15, wherein the process further comprises, after step (b), and preferably after step (b2), at least one pressing step (e) and / or at least one filtration step (c).

17. A process according to any one of claims 9 to 16, wherein the process, when step (b) is carried out in the presence of at least one aqueous, alcoholic or hydro-alcoholic solvent, further comprises, after step (b), at least one solvent removal step (h), preferably by evaporation.

18. Aqueous, hydro-alcoholic or alcoholic extract of at least one plant of the genus Perilla, characterized in that it is capable of being obtained by a process according to any one of claims 9 to 17.

19. Non-therapeutic use, in particular cosmetic use, of at least one extract according to any one of claims 1 to 8, to prevent and / or combat skin aging, in particular photo-induced skin aging, improve cell longevity, restore the skin barrier function and / or improve skin hydration.

20. Use according to the preceding claim, wherein the extract is administered orally or topically, and preferably topically.

21. Composition, in particular cosmetic, pharmaceutical, nutraceutical or food, and preferably cosmetic, comprising at least one extract according to any one of the preceding claims 1 to 8 or obtained by means of a process according to any one of claims 9 to 17, optionally in association with at least one physiologically acceptable medium.

22. Composition according to the preceding claim, comprising between 0.01% and 20%, preferably between 0.1% and 10%, more preferably between 0.5% and 5%, and particularly between 1% and 3%, by weight of extract(s), relative to the total weight of the composition.38 23. Non-therapeutic cosmetic treatment process, in particular for makeup and / or care, of a keratinous material, in particular of the skin, lips or hair, comprising at least one step of applying to said keratinous material at least one extract according to any one of claims 1 to 8 or a composition according to claim 21 or 22.

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