Compositions containing Sanguisorba officinalis root extract and uses thereof

Abstract

Compositions containing Sanguisorba officinalis root extract and uses thereof The present invention relates to a Sanguisorba officinalis root extract and its use as an active ingredient for skin and / or scalp care.

Description

[Technical Field]

[0001] The present invention relates to a root extract of the plant Sanguisorba officinalis for promoting light-related human mood. The present invention also relates to a root extract of Sanguisorba officinalis enriched with feruloylated tormentic acid derivatives and a method for preparing such an extract. Furthermore, the present invention relates to a composition comprising such a root extract, and to the use of such a root extract as an active ingredient for promoting light-related human mood in skin and / or scalp care. [Background technology]

[0002] Sunlight is essential for many organisms, including humans, and exerts several powerful biological effects beyond visual responses. It regulates circadian rhythms and brain activity, leading to the regulation of sleep and learning. Furthermore, sunlight directly affects human mood without causing circadian arrhythmias or sleep disorders (LeGates et al., Nature Reviews Neuroscience, Vol. 15, pages 443-454, 2014). It is well documented that lack of light is associated with depressive symptoms and cognitive impairment. The retina is a key organ involved in light detection, and opsins are key phototransduction molecules found therein. As the organ most exposed to light, skin also expresses photoreceptors called opsins. In addition to the retina, opsins are expressed in the epidermis, dermis, and hair follicles. Various opsins have been described, each responsive to different wavelengths of light. In the eye, one of the most abundant opsins is peropsin (RRH), which is also expressed in human skin. The biological roles of opsins in skin physiology are still largely unknown, but some are associated with skin differentiation, circadian rhythms, skin matrix remodeling via matrix metalloproteinase (MMP) regulation, or wound healing stimulation (Suh et al., Photodermatol Photoimmunol Photomed, Vol. 36(5), pages 329-338, 2020). The skin's ability to sense and transduce light independently of the eye's involvement via its retina is achieved in humans by exposure to light. It has been demonstrated that shining light onto the back of the knee affects circadian rhythms, and UV-A exposure using opaque goggles increases serum serotonin, resulting in a more balanced, less nervous sensation.

[0003] Serotonin, the body's natural mood booster, is a key mediator of the bidirectional interaction between the neuroendocrine system and the skin. Human skin possesses a serotonergic system capable of producing serotonin. This places the skin in a central position in the link between light and good mood, where serotonin is crucial. Serotonin is also a precursor to melatonin, the "sleep hormone."

[0004] Melatonin is a neurohormone associated with the light / dark cycle. Light can suppress or synchronize melatonin production according to the light schedule, and melatonin can stabilize and strengthen the coupling of circadian rhythms. Exposure to daytime light has been shown to improve human sleep while stimulating morning melatonin levels without affecting evening melatonin levels. Melatonin is also known to be synthesized and metabolized in the skin, where it plays several roles. Some of its functions are related to its antioxidant properties, including protection against ultraviolet and X-ray radiation. In particular, melatonin is a potent protector against UV radiation (primarily UV-B radiation). Furthermore, melatonin also activates antioxidant cascades that reduce free radicals and DNA repair systems in the skin. Finally, melatonin enhances the skin barrier through stimulation of the expression of the involucrins keratin-10 and keratin-14, and promotes skin wound healing.

[0005] Another sunlight-related beneficial effect is the production of the essential nutrient vitamin D, often referred to as the "sunshine" vitamin. Skin is the human body's primary source of vitamin D after sun exposure. Unfortunately, approximately 75% of the world's population is deficient in vitamin D due to spending significant amounts of time indoors. Upon UV-B absorption, the precursor 7-dehydrocholesterol (7-DHC) is converted to vitamin D3 in the skin, a process accelerated by thermal energy. These reactions are nonenzymatic and depend on UVB dose and temperature. 7-DHC in keratinocytes and skin fibroblasts is converted to previtamin D. Vitamin D is enzymatically hydroxylated to 25-hydroxyvitamin D (25(OH)D) and then to its biologically active metabolite, 1,25(OH)2D. Vitamin D synthesized in the skin can be released from cell membranes and enter the systemic circulation bound to vitamin D-binding protein (DBP). Vitamin D produced in the skin provides over 90% of the body's vitamin D needs. Vitamin D status affects cognitive, behavioral, and mood disorders. Furthermore, people with vitamin D deficiency are more likely to have any type of sleep disorder, poorer sleep quality, shorter sleep duration, and / or excessive daytime sleepiness.

[0006] The skin itself can respond to the active metabolites of vitamin D3. The primary genomic and biological effects of vitamin D3 metabolites in the skin are mediated through their binding to the nuclear vitamin D receptor (VDR). Indeed, skin expresses the VDR, which serves as the site of vitamin D action. Activated by the classical 1,25(OH)2D3 receptor, the VDR induces rapid response signaling through a non-genomic, membrane-associated mechanism based on alternative ligand binding sites or through action on the 1,25D3-MARRS receptor. Via activation of the nuclear VDR, vitamin D can exert several distinct effects on primary skin cells (keratinocytes and fibroblasts) and immune cells. Vitamin D also plays an important role in skin homeostasis, contributing to its barrier function and favoring skin differentiation. Its deficiency is associated with many proliferative and inflammatory skin disorders. Furthermore, as an essential part of a functioning immune system, the active form of vitamin D can regulate skin immunity and exert antioxidant, antifibrotic, and anti-inflammatory properties.

[0007] Sunlight is also beneficial for maintaining a balanced circadian rhythm, which underlies the regulation of a wide range of cellular, metabolic, physiological, and behavioral activities in mammals. The master clock is greatly influenced not only by light but also by the environment. Focusing on light, its effect on the phase of the circadian clock depends on the timing of light exposure. In this regard, several important circadian genes have been identified in the skin.

[0008] Because of these proven beneficial effects of light, phototherapy or photobiomodulation is used both for the treatment of mood disorders, as well as for the treatment of various skin conditions such as psoriasis, atopic dermatitis, hair regrowth, wound healing, and tissue regeneration.

[0009] On the other hand, sunlight also has harmful effects on the skin called photodamage, such as hyperpigmentation, melanoma due to DNA damage, and premature aging, and weakens mechanical properties, i.e., degradation of extracellular matrix (ECM) components, dehydration, and changes in skin complexion, resulting in loss of elasticity and firmness. It has been shown that broad protection against the entire solar spectrum, including UV-B, UV-A, visible light, and short-infrared rays, is necessary to prevent sun-related skin damage.

[0010] Carbonylated proteins (CPs) are synthesized by reactions between amino groups in proteins and reactive aldehyde compounds resulting from lipid peroxidation initiated by reactive oxygen species. In skin, CPs are detected more frequently in sun-exposed areas of elderly subjects [1]. CPs have also been detected in stratum corneum (SC) samples taken from skin in winter, even though sunlight radiant energy is weaker at that time [2]. CPs have been reported to correlate with decreased skin moisture and transepidermal water loss (TEWL), changes in the skin matrix, increased darkness around facial pores, and changes in skin color, such as a yellowish-dark color that alters skin tone (Masaki et al., J. Dermatol Science, Vol. 84, 1: pages 05-16 [01-04], 2016).

[0011] Sanguisorba officinalis, also known as great burnet or Sanguisorbe officinale, is a perennial herbaceous plant in the Rosaceae family, found in the Northern Hemisphere, Europe, Asia, and North America.

[0012] Root extracts of Sanguisorba officinalis are known to contain metabolites such as ellagitannins, flavonoids, and polyphenols. These extracts have been reported to have antiviral, antibacterial, hemostatic, anti-inflammatory, and anticancer activities, with diglycosides I and II and sanguiin H-6 only partially accounting for these activities (Jang et al., A Review, The American Journal of Chinese Medicine, Vol. 46, No. 2, pp. 1–20, 2018).

[0013] Sanguisorba officinalis root extract also contains euscaphic acid and / or tormentic acid(s) (Seongdae et al., Molecules, 23, 3001, 2018) (Non-Patent Document 5), ferulic acid or its esters. Ferulic acid exhibits antioxidant and anti-tyrosinase activity.

[0014] Tormentic acid belongs to the triterpenoid group of compounds with six isoprene units. It is known to have anti-cancer, anti-atherosclerotic, anti-inflammatory, anti-diabetic, antimicrobial, cardioprotective, and neuroprotective properties. Tormentic acid is widely found in many plants, but it also exists in various derivative forms that are more specific to certain plant families and certain tissues. While euscaphic acid, or the glycosylated form of tormentic acid, is the most common, these forms often have unique activities that differ from the basic form (tormentic acid).

[0015] Feruloylated derivatives of tormentic acid are rare natural molecules that may be of interest in the cosmetic field. In particular, 3-O-trans-feruloyl tormentic acid has been found only in extracts of the plants Vitex rotundifolia and Vitex trifolia, and 3-O-trans-feruloyl euscaphic acid has been found only in extracts of the leaves of Eriobotrya japonica. Another compound, oryzanol, has a structure similar to ferulic acid combined with a triterpene (but not tormentic acid) and was not toxic in mice. This suggests that these molecules may be of interest in cosmetics.

[0016] It has been discovered that the roots of Sanguisorba officinalis, unlike the aerial parts, may contain ferulic acid linked to tormentic acid via an ester bond. Therefore, access to the roots of Sanguisorba officinalis is key to recovering these compounds of interest. However, such manipulation involves the total or partial destruction of the plant, which is undesirable in light of the Nagoya Protocol and international standards (ISO 26000) guidelines for controlling access to biodiversity and ensuring sustainable development and social responsibility.

[0017] The use of Sanguisorba officinalis extracts is known in cosmetics. For example, CN-B 105193680 (Patent Document 1) and KR-A 20190003011 (Patent Document 2) disclose Sanguisorba officinalis root extracts for promoting skin-whitening properties. According to CN-B 105193680, the root extract is obtained using a resin fractionation step, during which many active ingredients, including feruloylated tormentic acid derivatives, are reliably adsorbed into the resin. EP-B1 0993826 (Patent Document 3) discloses Sanguisorba officinalis root extracts for stimulating melanin production in the skin. JP-B 3449967 (Patent Document 4) discloses Sanguisorba officinalis root extracts for stimulating collagen synthesis after UV-B exposure. WO-A1 2018000060 (Patent Document 5) discloses a root extract of Sanguisorba officinalis as a sebum regulator for oily skin. EP-A2 1051965 (Patent Document 6) discloses a root extract of Sanguisorba officinalis for improving ceramide production and thus enhancing skin moisturizing effects.

[0018] Diglycoside I (a triterpenoid saponin) isolated from the ethanol extract of the roots of Sanguisorba officinalis has been reported to exhibit anti-inflammatory and anti-wrinkle activities (Young Heui KIM et al., Bioscience, Biotechnology and Biochemistry, 72:2, 303-311, 2014, DOI:10.1271 / bbb.70268) (Non-Patent Document 6).

[0019] Since the majority of the world's population spends most of their time indoors, there is an unmet demand for natural products that can be used to at least partially mimic the beneficial effects that sunlight can bring to the body, preferably the skin, even in the absence and / or weak sun exposure conditions.Furthermore, it has been observed that an increasing number of cosmetic consumers are looking for natural products that can meet two criteria.The first criterion concerns the visual effect observed after application of these products.The second criterion concerns the emotional aspect, in particular the happiness that users can feel after application of these products. [Prior art documents] [Patent documents]

[0020] [Patent Document 1] CN-B 105193680 [Patent Document 2] KR-A 20190003011 [Patent Document 3] EP-B1 0993826 [Patent Document 4] JP-B 3449967 [Patent Document 5] WO-A1 2018000060 [Patent Document 6] EP-A2 1051965 [Non-patent literature]

[0021] [Non-Patent Document 1] LeGates et al.,Nature Reviews Neuroscience,Vol.15,pages 443-454, 2014 [Non-patent document 2] Suh et al.,Photodermatol Photoimmunol Photomed,Vol.36(5),pages 329-338,2020 [Non-patent document 3] Masaki et al.,J.Dermatol Science,Vol.84,1: pages 05-16[01-04],2016) [Non-patent document 4] Jang et al.,A Review, The American Journal of Chinese Medicine,Vol.46,No.2,1-20,2018 [Non-Patent Document 5] Seongdae et al., Molecules, 23, 3001, 2018 [Non-patent document 6] Young Heui KIM et al., Bioscience, Biotechnology and Biochemistry, 72:2, 303-311, 2014, DOI:10.1271 / bbb.70268 Summary of the Invention [Problem to be solved by the invention]

[0022] Therefore, there is an unmet need to provide a product of natural origin, namely a root extract of Sanguisorba officinalis, which can have a visible beneficial effect on the body, in particular on the skin and / or scalp, and also exhibit a feeling of well-being in the subject.

[0023] Sunlight can also have well-known harmful effects on the skin. Therefore, there is also an unmet need for skin care products of natural origin that preferably prevent light-related skin damage and thus can delay the signs of photoaging. [Means for solving the problem]

[0024] Surprisingly, it has been found that a root extract of Sanguisorba officinalis can mimic sunlight-related effects on the body, preferably the skin, even in the absence and / or under weak light exposure conditions. In this regard, the root extract of Sanguisorba officinalis can be used, for example, in phototherapy, to obtain light-related beneficial effects on the body, and thus a sense of relaxation, to improve human mood, even in the absence and / or under weak light exposure conditions.

[0025] Surprisingly, it has also been found that the root extract of Sanguisorba officinalis can be enriched with some interesting components that may be naturally present in small amounts. Indeed, the root of Sanguisorba officinalis can be enriched, for example, with feruloylated derivatives of tormentic acid by cultivating the plant under specific conditions without damaging the development of the whole plant. The enriched root extract can be advantageously used for skin and / or scalp care.

[0026] Summary of the Invention One aspect of the present invention relates to a Sanguisorba officinalis root extract comprising: tormentic acid, representing at least 1% by weight relative to the total weight of the dry extract; - feruloylated derivatives of tormentic acid having the general formula (I)

[0027] [ka] - feruloylated derivatives of deoxytormentic acid having the general formula (II):

[0028] [ka] The present invention further relates to a method for producing a root extract of the plant Sanguisorba officinalis, comprising the steps of: a) cultivating Sanguisorba officinalis under soil-free conditions, in particular aeroponic cultivation; b) stimulating the roots of the plant; c) solid / liquid extraction by maceration of the roots obtained in step b); d) recovering the extract obtained in step c), and e) Optionally, diluting and / or clarifying the extract recovered in step d) by successive filtration.

[0029] The present invention further relates to a cosmetic or dermatological or nutraceutical composition comprising the Sanguisorba officinalis root extract as defined above, and optionally one or more excipients, which are preferably cosmetically or dermatologically or nutraceutical acceptable.

[0030] The present invention further relates to the (cosmetic) use of a Sanguisorba officinalis root extract as defined above as an active ingredient for skin and / or scalp care.

[0031] The present invention also relates to a non-therapeutic method for preventing or delaying the appearance of skin aging effects and promoting light-related mood in humans, which method comprises applying to at least one part of the body a Sanguisorba officinalis root extract as defined above.

[0032] Another aspect of the present invention relates to the (cosmetic) use of a root extract of Sanguisorba officinalis for promoting light-related human mood.

[0033] All documents cited or referenced herein ("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]

[0034] DESCRIPTION OF THE DRAWINGS Figures 1A and 1B show the stimulation of happiness hormone production in skin explants treated with Sanguisorba officinalis root extract according to one embodiment of the present invention. Figure 1A shows the concentration of serotonin released by skin cells in the culture medium after 3 days of treatment. Figure 1B shows the concentration of melatonin released by skin cells in the culture medium after 5 days of treatment. 2A and 2B show clinical assessments related to improved well-being in volunteers treated topically with Sanguisorba officinalis root extract according to one embodiment of the present invention. Figure 2A demonstrates improved relaxation between days 1 and 5 after treatment. Figure 2B demonstrates improved relaxation at day 7 after treatment. FIG. 3 shows an automated assessment questionnaire related to well-being effects for a panel of 36 volunteers treated topically with a Sanguisorba officinalis root extract according to one embodiment of the present invention. Figures 4A and 4B show the modulation of vitamin D downstream biological pathways in the skin by Sanguisorba officinalis root extract according to one embodiment of the present invention. Figure 4A shows the regulation of vitamin D receptor. Figure 4B shows the regulation of vitamin D binding protein. FIG. 5 shows the induction and potentiation of phototransduction signals on skin explants treated with tormentic acid and feruloylated tormentic acid derivatives isolated from Sanguisorba officinalis root extract according to one embodiment of the present invention. FIG. 6 shows the improvement in skin elasticity observed during a clinical trial conducted on volunteers who were treated topically with a Sanguisorba officinalis root extract according to one embodiment of the present invention. FIG. 7 shows the improvement in skin complexion observed during a clinical trial conducted on volunteers who were treated topically with a Sanguisorba officinalis root extract according to one embodiment of the present invention. FIG. 8 shows the carbonylated protein content in keratinocytes of volunteers treated with a root extract of Sanguisorba officinalis according to one embodiment of the present invention, 28 days after application.

[0035] The elements of the present invention will now be described in more detail. While these elements are listed with specific embodiments, 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 elements described in this application should be deemed to be disclosed by the description of this application, unless the context dictates otherwise.

[0036] 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. As 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.

[0037] 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.

[0038] Detailed Description of the Invention One aspect of the present invention relates to a Sanguisorba officinalis root extract comprising: tormentic acid, representing at least 1% by weight relative to the total weight of the dry extract; - feruloylated derivatives of tormentic acid having the general formula (I)

[0039] [ka] - feruloylated derivatives of deoxytormentic acid having the general formula (II):

[0040] [ka] According to one embodiment, tormentic acid represents from 1 to 10% by weight, preferably from 1.2 to 8% by weight, more preferably from 1.5 to 7% by weight, in particular from 1.8 to 5% by weight, relative to the total weight of the dry extract.

[0041] According to one embodiment, the feruloylated derivatives of tormentic acid represent at least 0.1% by weight relative to the total weight of the dry extract. Advantageously, the feruloylated derivatives of tormentic acid represent between 0.1 and 5%, preferably between 0.2 and 4.5%, more preferably between 0.3 and 4%, even more preferably between 0.4 and 3.5% and in particular between 0.5 and 3% by weight relative to the total weight of the dry extract.

[0042] According to one embodiment, the feruloylated derivative of deoxytormentic acid represents at least 0.05% by weight relative to the total weight of the dry extract. Advantageously, the feruloylated derivative of deoxytormentic acid represents between 0.05 and 3%, preferably between 0.06 and 2.8%, more preferably between 0.07 and 2.6%, even more preferably between 0.08 and 2.4%, most preferably between 0.09 and 2.2%, and in particular between 0.1 and 2% by weight relative to the total weight of the dry extract.

[0043] In a preferred embodiment, the feruloylated derivative of tormentic acid represents at least 0.1% by weight and the feruloylated derivative of deoxytormentic acid represents at least 0.05% by weight, all weights being relative to the total weight of the dry extract.

[0044] According to one embodiment, tormentic acid, its feruloylated derivatives and its deoxytormentic acid feruloylated derivatives represent in total at least 1.15% by weight relative to the total weight of the dry extract. Advantageously, tormentic acid, its feruloylated derivatives and its deoxytormentic acid feruloylated derivatives represent in total from 1.15 to 18% by weight, preferably from 1.30 to 15% by weight, even more preferably from 1.40 to 12% by weight and in particular from 1.60 to 10% by weight relative to the total weight of the dry extract.

[0045] According to one embodiment, the root extract may further contain one or more ellagitannins.

[0046] According to one embodiment, the root extract may be in liquid form and may contain a solvent selected from water, lower alcohols, glycols or mixtures thereof, and dicaprylyl ether. The lower alcohol may preferably be selected from methanol and ethanol. The glycol may preferably be selected from dipropylene glycol, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, pentylene glycol, and glycerol. Preferably, the solvent may be selected from 1,3-propanediol, 1,3-butanediol, and glycerol. More preferably, the solvent is 1,3-propanediol. Particularly preferably, the solvent is bio-based 1,3-propanediol.

[0047] Thus, the liquid root extract corresponds to the crude liquid extract obtained after step c) of solid / liquid extraction by (optionally stimulated) root maceration of the roots, and also after step d). 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 content of feruloylated derivatives of tormentic acid and deoxytormentic acid.

[0048] According to one embodiment, 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 example for placing the root extract in liquid form in a hot, dry atmosphere in order to evaporate the maceration solvent.

[0049] The solid or viscous root extract can be further diluted in a dilution solvent to obtain another type of liquid root extract. Thus, the "dilution solvent" is a solvent used to dilute the solid or viscous root extract obtained. Advantageously, such a root extract contains tormentic acid, a feruloylated derivative of tormentic acid, and a feruloylated derivative of deoxytormentic acid. The dilution solvent can be selected from alcohols, 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 content of the feruloylated derivative of tormentic acid and the feruloylated derivative of deoxytormentic acid, a specific solvent must be used as a maceration solvent, but other solvents may also be used to dilute the solid or viscous root extract obtained before use. In particular, the dilution solvent does not have to be the same as the maceration solvent used to obtain the extract of the invention, and thus, said dilution solvent can preferably be chosen from methanol, ethanol, 1,3-propanediol, pentylene glycol, glycerol, dicaprylyl ether, in particular bio-based dicaprylyl ether.

[0050] Depending on whether the root extract is in liquid form, 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 described above or the dilution solvent described above.

[0051] "Tormentic acid" refers to tormentic acid itself as well as its stereoisomers.

[0052] "Feruloylated derivative of tormentic acid" means a compound having the following general formula (I) corresponding to the molecular formula: C40H56O8.

[0053] [ka] "Feruloylated derivative of deoxytormentic acid" means a compound having the following general formula (II), corresponding to the molecular formula: C40H56O7.

[0054] [ka] "Roots enriched in feruloylated derivatives of tormentic acid" and "enriched root" refer to a root extract of said plant that contains a higher amount of at least one feruloylated derivative of tormentic acid (Formula (I) or Formula (II)) compared to a corresponding root extract of the same plant as can be found in nature.

[0055] "Feruloylated derivatives of tormentic acid" means feruloylated derivatives of tormentic acid and feruloylated derivatives of deoxytormentic acid.

[0056] Preferably, to obtain a Sanguisorba officinalis root extract according to one aspect of the present invention, when the roots are deemed sufficiently developed, they are contacted with an extraction solvent by soaking or, preferably, maceration, followed by recovering and processing said extraction solvent to extract therefrom secondary metabolites containing feruloylated derivatives of tormentic acid released by the roots. This preferred method is marketed by a company named Plant Advanced Technologies (PAT) under the trade name "PAT Plantes a Traire" (PAT Plantes a Traire). (登録商標) " and is adapted from the method described in international application WO 01 / 33942. The teachings of this document are incorporated into the present specification by reference.

[0057] Therefore, the present invention further relates to a method for producing a root extract of the plant Sanguisorba officinalis according to one aspect of the present invention, the method comprising the steps of: a) cultivating Sanguisorba officinalis under soil-free conditions, in particular aeroponic cultivation; b) stimulating the roots of the plant; c) solid / liquid extraction by root maceration of the roots obtained in step b); d) recovering the extract obtained in step c), and e) Optionally, diluting and / or clarifying the extract recovered in step d) by successive filtration.

[0058] In one embodiment, the Sanguisorba officinalis root extract according to one aspect of the present invention can be obtained by the method described above.

[0059] "Soilless cultivation" refers to any cultivation method in which plant root development does not occur in soil. More precisely, soilless 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.

[0060] Various soil-free cultivation techniques are known, such as substrate-free systems requiring oxygen-enriched nutrient solutions, 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 practice, roots are nourished by a nutrient mist obtained by atomizing the nutrient solution in a closed medium. Substrate-based systems include subirrigation, in which the nutrient solution percolates through the bottom of the substrate, 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.

[0061] The terms "aeroponic plant cultivation" and "aeroponic" refer 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.

[0062] 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 to obtain maximum root growth and maximum production of secondary metabolites (including feruloylated derivatives of tormentic acid having Formula I and / or Formula II).

[0063] 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 (including feruloylated derivatives of tormentic acid having Formula I and Formula II).

[0064] According to a preferred embodiment, plants may be nourished using a nutrient solution mist obtained by atomizing the nutrient solution in a closed system medium through a mister.

[0065] According to one embodiment, the plants can be placed on trays with the aerial parts of the plants above the tray and the root parts below the tray, the trays being placed on a table forming a holding area for collecting excess liquid diffused towards the plants, and the trays being transferred 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 also incorporated herein by reference.

[0066] In a preferred embodiment, during step a), aeroponic Sanguisorba officinalis is nutrient-supplemented by spraying the roots with a nutrient solution of essential mineral salts (nitrogen - N, phosphorus - P, potassium - K) to maximize root growth and the concentration of the secondary metabolites mentioned above without harming plant survival. A skilled artisan will know by common sense how to adapt the proportions and concentrations of various mineral salts to optimize plant growth, and especially root growth. In this case, the mineral salt concentration of the nutrient solution is typically within the low conductivity range, advantageously ranging from 0.4 to 1.6 mS / cm, preferably 0.8 to 1.2 mS / cm, to promote a greater diversity of secondary metabolites, including tormentic acid and its feruloylated derivatives, in the root extract.

[0067] Advantageously, the above aeroponic cultivation conditions allow obtaining quantities of several secondary metabolites, including feruloylated derivatives of tormentic acid, superior to the quantities of these same compounds obtained in soil cultivation.

[0068] In a preferred embodiment, the method may include a root stimulation step. In this case, the root stimulation step b) of the plant includes placing the roots in contact with a nitrogen-deficiency stimulation nutrient solution, which may contain a nitrogen ratio less than that generally considered optimal for plant growth, particularly root growth. Advantageously, the nutrient solution may contain less than 15% nitrogen, more preferably no nitrogen, and steps a) and b) may be sequential or simultaneous.

[0069] In a preferred embodiment, root stimulation of the plant in step b) comprises contacting the roots with a nitrogen-deficient nutrient solution. Placing the plant in contact with the nitrogen-deficient nutrient solution causes "nitrogen stress", which is responsible for stimulating the production of secondary metabolites, in particular feruloylated derivatives of tormentic acid.

[0070] In a preferred embodiment, the nitrogen deficiency stimulating nutrient solution is typically 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.

[0071] In one embodiment, the root stimulation step b) significantly increases the content of secondary metabolites in the roots, in particular feruloylated derivatives of tormentic acid, and thus promotes the flux of said metabolites from the roots into the solvent selected for extraction, and makes it possible to return the plant to cultivation and then reuse it, without completely losing the viability of the plant. In other words, the plant stimulation step promotes the biosynthesis of said secondary metabolites.

[0072] According to one embodiment, root stimulation in step b) can be performed by feeding the roots with a nitrogen-deficient N / P / K stimulated nutrient solution vaporized or misted onto the roots.

[0073] According to one embodiment, step b) can be carried out by spraying or macerating the roots with an N / P / K nutrient solution containing less than 6%, more preferably less than 3% nitrogen, said solution preferably being evaporated or misted above the roots.

[0074] According to one embodiment, step b) of stimulating the roots by feeding them with a nitrogen-deficient N / P / K nutrient solution vaporized on said roots is advantageously carried out for a period of between 1 week and 8 weeks, more particularly between 1 week and 3 weeks, preferably for a period of 2 weeks.

[0075] According to one embodiment, during step b), the mineral salt concentration of the nutrient solution may be in the low conductivity range, advantageously ranging from 0.4 to 1.2 mS / cm, preferably from 0.6 to 1.0 mS / cm, to promote a greater diversity of secondary metabolites, including feruloylated derivatives of tormentic acid.

[0076] According to one embodiment, step b) and step a) may be carried out simultaneously, and the nutrient solution may then be replaced by the stimulating solution.

[0077] According to a preferred embodiment, the method for preparing the root extract of the plant Sanguisorba officinalis of the present invention comprises: a) cultivating Sanguisorba officinalis under soil-free conditions, in particular aeroponic cultivation; b) stimulating the roots of the plant; c) solid / liquid extraction by maceration of the roots obtained in step b); d) recovering the extract obtained in step c), and e) Optionally, diluting or clarifying the extract recovered in step d) by successive filtration.

[0078] Subsequently, the cultivated and stimulated plants are typically subjected to step c) of solid / liquid extraction by root maceration under defined conditions with respect to solvent, temperature and extraction time, thereby ensuring that a root extract enriched in feruloylated derivatives of tormentic acid, together with other secondary metabolites, is obtained. Solid / liquid extraction is a known solvent-based extraction technique. The secondary metabolites released by the roots are recovered in the extraction solvent.

[0079] According to a preferred embodiment, step c) of solid / liquid extraction of the roots stimulated during step b) may be 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 immersion step.

[0080] According to one embodiment, root maceration can be carried out on cut, dried, and optionally pulverized roots. The drying method can be any suitable drying method known in the art, in particular by subjecting the root biomass to temperatures between 30°C and 60°C for 24 hours 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 known method, 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 root growth and promote the root production of secondary metabolites.

[0081] According to a preferred embodiment, the method may comprise the step of cutting the roots and drying the cut roots prior to the step of maceration, the maceration being carried out by placing the cut and dried roots in contact with a solvent.

[0082] According to a preferred embodiment, step c) of the solid / liquid extraction may involve placing the roots in contact with a solvent selected from water, alcohol, glycol, or a mixture thereof, and dicaprylyl ether, preferably bio-based dicaprylyl ether. The alcohol may be selected from ethanol and methanol, preferably used pure or in the form of an aqueous solution containing 10% to 99.9%, more preferably 40% to 90%, and especially 50% to 85% alcohol. The glycol may be selected from dipropylene glycol, propane-1,3-diol, propane-1,2-diol, pentylene glycol, 1,3-butanediol, and glycerol, pure or in the form of an aqueous glycol solution containing 10% to 99.9%, preferably 70% to 99.9% glycol.

[0083] Preferably, the solvent is a glycol, such as 1,3-propanediol, 1,3-butanediol, or glycerol. In a preferred embodiment, the solvent is selected from 1,3-propanediol. Particularly preferably, the solvent is bio-based 1,3 propanediol.

[0084] According to one embodiment, step c) may comprise contacting the roots with the solvent, e.g., by maceration, for a time period between 30 minutes and 48 hours, preferably between 1 hour and 24 hours. In such cases, the operating temperature may be between 20°C (room temperature) and 80°C, preferably between 40°C and 60°C.

[0085] According to a particular embodiment, for root maceration of dried roots, the ratio of the amount of dry root to the amount of solvent can range between 1 kg dry root / 10 kg solvent to 1 kg dry root / 100 kg solvent, more preferably between 1 kg dry root / 20 kg solvent to 1 kg dry root / 40 kg solvent.

[0086] In one embodiment of the present invention, "root maceration in a solvent" refers to root maceration for which the solvent placed in contact with the roots can be, for example, ethanol used pure or in aqueous solution form (the aqueous solution contains between 50% and 85% ethanol), dicaprylyl ether used pure, 1,3-butanediol used pure or in aqueous solution form (the aqueous solution contains between 50% and 85% 1,3-butanediol), or 1,3-propanediol used pure or in aqueous solution form (the aqueous solution contains between 50% and 85% 1,3-propanediol). More preferably, the solvent is bio-based 1,3-propanediol used pure or in aqueous solution form (the aqueous solution contains between 50% and 85% bio-based 1,3-propanediol).

[0087] According to one embodiment, the method may comprise one or more additional steps of processing the plant root extract, which may be selected from the following known methods: - Dilution, concentration, - one or more filtrations, in particular a clarifying filtration and / or a sterile filtration, - solid / liquid extraction, - refining, -Bleaching liquid root extract.

[0088] The present invention further relates to a Sanguisorba officinalis root extract obtainable by the method as detailed above.

[0089] The present invention further provides the following: - a Sanguisorba officinalis root extract according to the above aspects of the invention or a Sanguisorba officinalis root extract obtained by the method defined above; and - at least one cosmetically or dermatologically acceptable ingredient other than said extract of Sanguisorba officinalis, wherein the cosmetic or dermatological composition is a composition for topical use or an orally administered nutraceutical composition selected from the group consisting of a solution, suspension, emulsion, cream, paste, gel, lotion, powder, soap, surfactant-containing water, oil, shampoo, and spray.

[0090] According to a preferred embodiment of the composition, the root extract of Sanguisorba officinalis may represent 0.0001 to 15% by weight, preferably 0.001 to 10% by weight, more preferably 0.01 to 5% by weight, relative to the total weight of the cosmetic or dermatological or nutraceutical composition.

[0091] 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 suitable cosmetic treatment for a 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 in the art for cosmetic use, such as thickeners, preservatives, fragrances, dyes, chemical or mineral filters, moisturizers, thermal waters, etc.

[0092] Thus, the cosmetic composition of the present invention may further comprise other cosmetically active agents other than the Sanguisorba officinalis root extract defined above, such as other anti-ageing 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.

[0093] Suitable cosmetic excipients are known 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.

[0094] The cosmetic composition of the invention is therefore advantageously intended for external application and may in particular be in the form of a cream, milk, lotion, gel, serum, spray, mousse, solution, ointment, emulsion, patch or mask.

[0095] Suitable dermatological excipients are also known in the art, for example, they may be the same as those intended for use in cosmetics.

[0096] Suitable nutraceutical excipients are known in the art.Examples may 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, for example, a food supplement in the form of a solid, coated or uncoated tablet, liquid, powder, soft or hard gelatin capsule, etc.

[0097] It will be understood that the definitions and preferred embodiments made in the context of the Sanguisorba officinalis root extract according to one aspect of the present invention apply mutatis mutandis to compositions comprising said extract.

[0098] The present invention also relates to the (cosmetic) use of a root extract of Sanguisorba officinalis according to one aspect of the invention as defined above as an active ingredient for skin care and / or scalp care, in particular for promoting light-related human mood by stimulating the production of at least one well-being hormone in skin cells.

[0099] Advantageously, the Sanguisorba officinalis root extract according to one aspect of the invention as defined above is suitably useful in the context of the new lifestyle concept of well-aging or healthy aging, since said extract is able to stimulate well-being hormones and prevent the harmful effects of exogenous (e.g. UV rays) and endogenous factors on the skin, thus influencing the overall skin homeostasis and affecting both health and beauty.

[0100] Therefore, the Sanguisorba officinalis root extract of the present invention can reproduce or mimic the beneficial effects of light, preferably sunlight, on the body, preferably skin, even in the absence or low light exposure conditions. These light-related beneficial effects are obtained even in the absence or low light exposure conditions. In fact, the beneficial effects are even better under light exposure conditions.

[0101] According to one embodiment, the use can be for promoting the production of at least one happiness hormone in skin cells, for promoting or stimulating the expression of light transducting molecules, preferably peropsin (RRH), in skin cells, for maintaining or restoring the circadian rhythm of clock proteins in skin cells, for stimulating vitamin D downstream biological pathways, or a combination thereof.

[0102] Thus, the use for skin care can additionally be for preventing or delaying the signs of photoaging in the skin, for preventing or treating UV-related skin damage, or a combination thereof.

[0103] According to one embodiment, the use for skin care may be for reducing and / or preventing protein carbonylation in skin cells.

[0104] According to one embodiment, preventing or delaying the signs of photoaging on the skin can be for improving the mechanical properties of the skin, in particular for improving skin elasticity, for improving skin complexion, for increasing dermal density, for maintaining or restoring skin cell integrity, for maintaining or restoring skin barrier function, for promoting skin hydration, for improving skin tone, for preventing the formation of fine lines and wrinkles, or a combination of two or more thereof.

[0105] As used herein, the terms "well-being hormone" or "good mood hormone" or "happiness hormone" may be understood interchangeably in the broadest sense generally understood in the art as a molecule (which may be, for example, a hormone consisting of a protein or peptide, or a chemical molecule) capable of providing a happy or soothing effect in a subject.

[0106] "Weak light exposure conditions" in the overall context of the present invention means that the light intensity received by the body, skin or scalp can be greater than 5 lux, preferably greater than 200 lux, and even more preferably greater than 5000 lux.In contrast, for example, during phototherapy, the light intensity can generally be greater than about 5000 lux, preferably greater than about 10000 lux.

[0107] Advantageously, the light can be any light, which may be of natural origin such as sunlight, daylight, etc., or it may be artificial light emitted by any light source, for example emitted by a lamp, etc. Preferably, the light is sunlight or daylight.

[0108] In the overall context of the present invention, "beneficial effects associated with light" refers to, for example, health promotion, mood control, and feelings of relaxation. These effects can be obtained, for example, by measuring the expression levels of serotonin and melatonin in plasma and peropsin (RRH) in skin cells. It can also include circadian rhythm regulation, brain activity promotion, and sleep regulation. It also refers to increasing the activity of vitamin D in skin cells to meet the body's needs.

[0109] "Downstream biological pathway" in the general context of the present invention means the transport of the active form of vitamin D by its nuclear receptor (VDR: Vitamin D Receptor) and its carrier (DBP: Vitamin D Binding Protein) to exert its biological effect on the body. Thus, vitamin D can be distributed from the skin and / or scalp to organs throughout the body thanks to DBP.

[0110] It will be understood that definitions and preferred embodiments made in the context of the use of a root extract of Sanguisorba officinalis according to one aspect of the present invention apply mutatis mutandis to the use of said root extract of Sanguisorba officinalis. In particular, the term "active ingredient" may be understood in the broadest sense throughout the context of the present invention as an ingredient that may exhibit the desired and intended activity alone or together with one or more carriers that are inactive themselves.

[0111] The present invention also relates to a non-therapeutic method for preventing or delaying the appearance of skin ageing effects and / or promoting a sense of well-being and / or relaxation, comprising applying to at least a part of the skin and / or scalp a Sanguisorba officinalis root extract according to the above aspects of the invention. [Example]

[0112] example In the following examples, the root extract of Sanguisorba officinalis according to one embodiment of the invention, in which the plants are grown under soilless conditions, in particular aeroponic, is referred to as SORE.

[0113] SORE was prepared from plants whose seeds were purchased from a UK supplier (Seedaholic) between 2015 and 2018. SORE was prepared from plants whose seeds were purchased from a French supplier ("Les Semences du Puy") between 2019 and 2022. From 2019 onwards, Sanguisorba officinalis plants were kept and propagated by the applicant.

[0114] Example 1: Preparation of SORE and its characterization The SORE was obtained according to the following process: a) cultivating Sanguisorba officinalis aeroponically in a nutrient medium with an N / P / K composition equal to 15 / 10 / 30 respectively and a conductivity between 0.4 and 1.6 mS / cm for a period between 2 and 6 weeks; b) stimulating the plants with nitrogen stress for 1 to 3 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 by maceration of the dried roots in a solution of pure 1,3-propanediol at a temperature of 50°C for 2 to 24 hours; d) recovering the root extract obtained during step c); e) Clarifying by filtration.

[0115] For the purposes below, the feruloylated derivatives of tormentic acid having general formula I and formula II will be referred to as FDTA-I and FDTA-II, respectively. Tormentic acid will be referred to as TA.

[0116] Quantification of the amounts of TA and FDTA-I and FDTA-II in the SORE was performed according to the protocol described in Example 2.

[0117] - Dry extract content: 9.9g / kg~13.4g / kg - TA content: 119.9mg / Kg~217.2mg / Kg (i.e. 1.21%~1.67% of the dry extract) - FDTA-I content: 30.21mg / Kg~45.42mg / Kg (i.e. 0.3%~0.35% of the dry extract) - FDTA-II content: 7.32mg / KgL~11.90mg / Kg (i.e. 0.07%~0.09% of the dry extract)

[0118] The resulting SORE had 9 g to 13.4 g of dry extract per kilogram of extract and contained approximately 2 wt% of total markers (FTA-I and FTA-II) based on the total weight of the dry extract.

[0119] Example 2: Quantification method for FDTA-I and FDTA-II All samples were analyzed using a UHPLC Shimadzu Nexera X2 system (Shimadzu, Japan) equipped with a PDA detector coupled to a mass spectrometer LCMS2020 (electrospray ionization in negative ion mode, m / z 100–1000) using a Kinetex EVO C18 reversed-phase column (150 mm × 2.1 mm, 2.6 μm, Phenomenex, USA) maintained at 40 °C during all analyses. The mobile phase consisted of ultrapure water (Mili-Q, Merck Millipore) + 0.1% formic acid (Carlo Erba, France) (phase A) and pure acetonitrile (Sigma-Aldrich Chemie, Germany) (phase B), delivered at 0.5 ml / min with a gradient of phase B as follows: 5–95% (0–10 min); 95% (10–13.5 min); 95–5% (13.5–13.55 min), 5% (13.55–15.1 min).

[0120] Quantification of FDTA-I and FDTA-II in different extracts of Sanguisorba officinalis was performed using a ferulic acid standard prepared at a concentration of 100 mg / L in an ethanol / water mixture (70 / 30, v / v). The content of FDTA-I and FDTA-II was expressed as the equivalent amount of ferulic acid in each extract sample.

[0121] In the context of this disclosure, the concentrations of FDTA-I and FDTA-II in Sanguisorba officinalis extracts, expressed in mmol / L, were determined by measuring the areas of the peaks corresponding to FDTA-I and FDTA-II on the HPLC chromatogram (330 nm) of the extract. The peak areas of FDTA-I and FDTA-II were then divided by the peak area of ​​a standard solution containing 0.515 mmol / L of ferulic acid and multiplied by the molar concentration of the ferulic acid standard (0.515 mmol / L). The molar concentration values ​​of FDTA-I and FDTA-II (expressed in mol / L) were converted to mass concentrations (expressed in g / L) by multiplying them by the molecular weights of both compounds.

[0122] Quantification of FDTA-I and FDTA-II was performed according to the following formula:

[0123]

number

[0124]

number

[0125] Example 3: Phytochemical analysis of FDTA-I and FDTA-II amounts in different extracts of Sanguisorba officinalis using different culture modes and in different tissues Sanguisorba officinalis plants were grown aeroponically for 4 weeks in a 15 / 10 / 30 (N / P / K) culture medium with a conductivity between 1.0 and 1.2 mS / cm, followed by a 2-week root stimulation step in a defined solution with an N / P / K composition corresponding to 0 / 15 / 40 and a conductivity between 1 and 1.2 mS / cm. Roots harvested before changing the culture medium corresponded to unstimulated roots. Additionally, commercially available dried roots of Sanguisorba officinalis, consisting almost exclusively of rhizomes, were purchased (Xuewen Tang Bozhou Swanf Commerce & Trade) to evaluate their phytochemical composition.

[0126] The roots and aerial parts were cut, harvested, dried in a ventilated oven at 50 °C for 48 h, and ground. For the preparation of each extract, 25 mg of ground roots in powder form or 25 mg of ground aerial parts in powder form were macerated in 0.5 mL of ethanol / water (70 / 30 v / v) with stirring for 1 h at ambient temperature. Samples were centrifuged and analyzed by HPLC-UV-MS.

[0127] The results are summarized in Table 1 below:

[0128] [Table 1]

[0129] It was observed that the root extract of aeroponic cultivated Sanguisorba officinalis contains both FDTA-I and FDTA-II.

[0130] Furthermore, the concentrations of FDTA-I and FDTA-II were strongly increased by the stimulation step: - 2.17 times with respect to FDTA-I between unstimulated and stimulated roots of plants grown under aeroponic conditions; - 10.66 times with respect to FDTA-II between unstimulated and stimulated roots of plants grown under aeroponic conditions. Interestingly, a decrease in the amount of TA was observed in stimulated roots compared with non-stimulated roots, suggesting that TA may be combined with ferulic acid upon stimulation, thus increasing FDTA-I and FDTA-II.

[0131] The study also confirmed that: - Aerial parts from Sanguisorba officinalis do not contain FDTA-I and FDTA-II, regardless of whether they are grown in soil or under aeroponic conditions. - FDTA-I and FDTA-II were not detected in commercially available dried roots (exclusively rhizomes) in the above protocol tests carried out.

[0132] These results highlight one of the advantages of cultivating Sanguisorba officinalis under aeroponic conditions.

[0133] Example 4: Extraction process of TA, FDTA-I and FDTA-II from SORE The extract was prepared from the roots of aeroponically grown Sanguisorba officinalis with a stimulation step according to Example 3. A) Comparison of different extraction solvents The different solvents chosen for the solid / liquid extraction by maceration according to step c) of the method for preparing the root extract are 70 / 30 (v / v) ethanol / water, pure water, 30 / 70 (v / v) or 50 / 50 (v / v) and 70 / 30 (v / v) propane-1,3-diol / water mixtures, pure propane-1,3-diol, 80 / 20 (v / v) 1,3-butanediol / water mixture and pure dicaprylyl ether.

[0134] The extraction rates of TA, FDTA-I, and FDTA-II were compared with those of ethanol extractions carried out under identical conditions as described in Example 3, except for the maceration duration (4 hours in this example). The content of TA, FDTA-I, and FDTA-II in each sample was determined according to the protocol described in Example 2. The results are shown in Table 2 below:

[0135] [Table 2]

[0136] A 70 / 30 ethanol / water mixture can be considered a reference solvent for those skilled in the art. The results shown in Table 2 demonstrate that pure propane-1,3-diol has the best extraction rate of TA, FDTA-I, and FDTA-II among all the solvents tested. Pure propane-1,3-diol was selected for the following examples.

[0137] B) Comparison of different solvent-independent extraction methods To improve the extraction of TA, FDTA-I, and FDTA-II in SORE, a comparison of different extraction methods was carried out using pure propane-1,3-diol as the maceration solvent. Except for the extraction temperatures (25°C and 50°C), the extractions were carried out under the same conditions as those described in Example 3, and lasted for 2, 4, and 24 hours.

[0138] The content of TA, FDTA-I and FDTA-II in each sample was determined according to the protocol described in Example 2. Extraction yields were calculated for 50°C or 24 hours of extraction.

[0139] Regarding the extraction temperature experiments, the extract used in reference (100) had the following characteristics: - Dry extract content: 7.06g / Kg - TA content: 105.85mg / Kg (i.e. 1.50% of the dry extract) - FDTA-I content: 40.10 mg / Kg (i.e. 0.57% of the dry extract) - FDTA-II content: 29.27 mg / Kg (i.e. 0.41% of the dry extract)

[0140] Regarding the extraction duration experiments, the extract used for reference (100) had the following characteristics: - Dry extract content: 13g / Kg - TA content: 217.20 mg / Kg (i.e. 1.67% of the dry extract) - FDTA-I content: 45.42 mg / Kg (i.e. 0.35% of the dry extract) - FDTA-II content: 10.64 mg / Kg (i.e. 0.08% of the dry extract)

[0141] The results are set forth in Tables 3 and 4 below:

[0142] [Table 3]

[0143] These results confirmed that the optimum temperature for extracting TA, FDTA-I and FDTA-II is 50°C, which allows obtaining the following advantages compared to 25°C: - Increased yield of FDTA-I: 3-fold increase. - Increased yield of FDTA-II: 4.5-fold increase. - TA yield increase: 1.25 times increase. - Increased yield of FDTA-I and FDTA-II: 1.7-fold increase.

[0144] [Table 4]

[0145] These results confirmed that the optimal duration for extracting TA, FDTA-I and FDTA-II is 24 hours, which allows obtaining the following advantages over 2 and 4 hours: - Increased yield of FDTA-I: 1.5 and 1.25-fold increase, respectively. - FDTA-II yield increase: 1.45 and 1.3-fold increase, respectively. - TA yield increase: 1.8 and 1.3 fold increase, respectively. - Increased yield of TA and FDTA-I and FDTA-II: 1.8- and 1.3-fold increase, respectively.

[0146] Example 5: SORE induces feelings of well-being and relaxation through upregulation of serotonin and melatonin expression Serotonin, the body's natural mood booster, also known as the "mood hormone" or "happiness hormone," has been shown to be upregulated by sunlight. Skin may play a central role in the link between light and mood, and serotonin may be crucial here. Therefore, experiments demonstrated that SORE can mimic phototransduction signals in the skin. We wondered whether topically applied SORE could upregulate serotonin expression without sunlight exposure. Therefore, we assessed serotonin in the culture medium of skin explants treated with 1% SORE for 3 days in the absence of light using a commercially available ELISA kit (Biovison, Abcam) according to the manufacturer's instructions. Interestingly, SORE was shown to significantly stimulate serotonin production by the skin explants by +55% (Figure 1A).

[0147] For transcriptome analysis, total RNA was extracted after 24 hours of tissue culture 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. To analyze the data, all statistically significantly regulated genes were analyzed using the DAVID bioinformatics resource. This tool identifies functionally regulated pathways from large gene or protein datasets.

[0148] The transcriptome data (Table 5) showed that SORE can upregulate the mRNA expression of 5-hydroxytryptamine receptor 3A, one of several receptors for 5-hydroxytryptamine (serotonin).

[0149] [Table 5]

[0150] Serotonin is a precursor to melatonin, which is linked to sleep quality and, in turn, well-being. Melatonin secretion is dependent on the light / dark cycle. Melatonin also stimulates antioxidant and DNA repair systems, and its precursor, serotonin, was upregulated by SORE. Therefore, we assessed melatonin in the culture medium of skin explants treated with SORE for 5 days in the absence of light using a commercially available ELISA kit (Abbexa) according to the manufacturer's instructions. Interestingly, SORE was shown to significantly stimulate melatonin production by the skin explants by +120 percent on day 5 (Figure 1B).

[0151] The happiness effects of SORE were then demonstrated at a clinical level in two clinical trials.

[0152] A gel cream containing 1% SORE (see Table 9) was used in a double-blind and vehicle-controlled clinical trial, adhering to the recommendations of the Declaration of Helsinki and the International Conference on Harmonization Good Clinical Practice guidelines as applicable to non-pharmaceutical studies.

[0153] - In the first study (proof-of-concept study): Ten Caucasian volunteers, aged 32-58 years, consisting of four women and six men, applied SORE-containing gel cream to the entire face twice daily for five days. In this study, the same volunteers also tested SORE-containing gel cream and a placebo. There was a five-day washout period between the two sessions of SORE-containing gel cream evaluation. The order of application of SORE-containing gel cream and placebo was randomly assigned to the volunteers.

[0154] - In the second study, 36 Caucasian women aged 30-50 years were enrolled. Volunteers enrolled in this study declared depression. Volunteers had a dull complexion and lack of luminosity on their face. Panelists were randomized into two groups: a placebo formula (21 volunteers) and a SORE formula (15 volunteers). They were asked to apply either the SORE-containing gel cream or the placebo to their entire face twice daily for 28 days.

[0155] To investigate the improvement in the volunteers' mood (happiness) in these two studies, a headset device from MyBrain Technologies was used, capable of measuring the electrical signals emitted by the brain during its activity. These brain electrical signals are measured on the surface of the head by a captor in the headset. A proprietary algorithm calculates a relaxation index (the higher the value, the more relaxed the volunteer). Measurements were performed at least one hour after application of the product to avoid the influence of sensitive characteristics of the formulation (fragrance, texture, etc.). Therefore, the observed effects are due to SORE itself.

[0156] For Study 1, volunteers were measured on day 0 (before use of the SORE-containing gel cream), and then 1 and 5 days after twice-daily application of the SORE-containing gel cream.

[0157] For Study 2, measurements were taken on day 0 and after 7 days of twice-daily application of the SORE-containing gel cream.

[0158] To normalize the values ​​for each volunteer's initial state, the % variation was calculated for each volunteer with respect to its initial state according to the following: (final value - day 0 value) / day 0 value * 100 Used topically on the face, formulated SORE, in contrast to placebo, improved relaxation in volunteers as soon as day 1 of use. Furthermore, this effect was maintained after five days of use. Figure 2A shows that SORE improved relaxation: +112% on day 1 and +97% on day 5, while placebo showed no improvement: +17% on day 1 and -50% on day 5.

[0159] The second study confirmed the well-being effects of SORE formulation observed in the first clinical trial. Indeed, relaxation parameters improved after 7 days of SORE formulation compared with placebo control. Figure 2B shows that SORE formulation significantly improved relaxation at 7 days (+279% # p<0.07), whereas placebo failed to improve (-123.6% ns). The difference in SORE effect was significant compared with placebo (* p<0.05). The in vitro effects on serotonin secretion and well-being were confirmed in vivo. Furthermore, volunteers' self-assessments confirmed the well-being effects of SORE formulation (Figure 3). Volunteers reported feeling more self-aware, energized, and had an improved sense of well-being. Furthermore, volunteers noticed that their skin looked more refreshed and revitalized, two signs of changes in depression.

[0160] Example 6: SORE upregulates vitamin D downstream biological pathways Since vitamin D cannot be produced by the skin in the absence of sunlight (especially UVB), we wondered whether SORE could upregulate the vitamin D downstream biological pathways, thereby enabling the effects of sunlight. In fact, the active form of vitamin D requires the VDR and carrier DBP (vitamin D binding protein DBP) to exert its biological effects. Vitamin D can be distributed from the skin to organs throughout the body thanks to the vitamin D binding protein (DBP). Vitamin D may also have direct beneficial effects in the skin via its nuclear receptor (VDR). Therefore, we evaluated the effects of SORE on the vitamin D downstream biological pathways (VDR and DBP) at both the transcriptome level in skin explants 24 hours after SORE treatment and at the proteome level in skin explants 5 days after SORE treatment with or without phototherapy exposure.

[0161] The associated effect of SORE (see Table 9) formulated at 1% on vitamin D binding protein (DBP) was examined in transcriptome analysis, as described in Example 5, and the effect of coupling to phototherapy on vitamin D receptor (VDR) was examined.

[0162] [Table 6]

[0163] Figure 4 shows the upregulation of VDR (+15%; Figure 4A) and DBP (+35%; Figure 4B) protein expression in skin explants treated simultaneously with light (without UV-B rays) and 1% formulated SORE (see Table 9) for 5 days. All these results confirmed that SORE can improve the distribution of vitamin D produced in the skin to other tissues through the upregulation of its carrier (DBP). Furthermore, the experiment demonstrated that SORE can improve the biological effects of vitamin D in the skin, as indicated by the upregulation of VDR.

[0164] Example 7: A mixture of FDTA-I and TA from SORE mimics the effects of light on the skin SORE was formulated into a gel cream at 0.75% by weight (Table 7). The biological effects of SORE compared to placebo were evaluated ex vivo in human skin explants from a 39-year-old Caucasian female donor by transcriptome analysis. Briefly, the formulation was applied at 2 mg / cm2 onto the skin explants. 2 The explants were cultured at 37°C and 5% CO2 for 24 hours.

[0165] [Table 7]

[0166] SORE induced phototransduction signals similar to visible light (but in the absence of light). Transcriptome studies such as those described in Example 5 showed that SORE induced the expression of photoreceptors in the skin that are known to be induced by visible light, confirming that SORE may be able to induce phototransduction even in the absence of light exposure. Indeed, visible light has been described to stimulate opsin photoreceptors, such as perospin (RRH) or rhodopsin, to transmit light signals. Transcriptome skin analysis showed enrichment of transcripts involved in phototransduction signals and the homeostasis of photoreceptors normally found in the retina (Table 8):

[0167] [Table 8]

[0168] - RRH plays a role in retinal pigment epithelial physiology by detecting light directly or by monitoring the concentration of photoreceptor-derived compounds. It is also expressed in skin, where it may be involved in phototransduction of short-wavelength violet light, suggesting that the skin may indeed possess photoreceptors capable of perceiving and distinguishing between various light qualities.

[0169] - OR56A5 has been described as being impaired in retinitis pigmentosa, a disease associated with vision loss. ORA5 is involved in UV light signaling.

[0170] - SAG binds to light-activated phosphorylated Rho and competes with G proteins for the same binding site on Rho, thereby terminating G protein-mediated Rho signaling, which prevents light-dependent degeneration of retinal photoreceptor cells.

[0171] The ability of SORE to induce phototransduction signals in skin explants without light exposure, similar to visible light, was demonstrated. In other words, SORE can induce the expected biological effects of light on skin without light.

[0172] Additional ex vivo experiments were performed (skin from a 41-year-old female donor) in the presence or absence of light using a commercially available phototherapy device (Dayvia White072). This device is specifically designed for exposure to people suffering from seasonal mood swings. Briefly, a gel cream containing 1% SORE or placebo (see Table 9) was applied at 2 mg / cm2 onto skin explants. 2 The explants were exposed to a phototherapy device. The device emitted 10,000 lux of light at a distance of 21 cm, without UV or infrared light. 10,000 lux is the recommended light intensity for phototherapy. Skin explants received a placebo or SORE and were then treated with the device's light for 20 minutes at a distance of 21 cm. Topically treated explants without exposure were also cultured for reference. Daily exposures were administered, as well as daily topical applications of placebo or SORE. Topical application was administered immediately after light exposure. The explants were cultured at 37°C and 5% CO2 for 5 days.

[0173] At the end of the experiment, the explants were formalin-fixed and then embedded in paraffin. Thin sections were processed for RRH immunofluorescence. Expression levels were assessed by fluorescence quantification using Image J. Figure 5 shows that, as expected, light upregulated RRH expression. Furthermore, SORE was able to stimulate RRH expression in the absence of light (+17%), indicating its ability to mimic the effects of light on this photoreceptor, suggesting that SORE may initiate a light-like effect. This result supported the transcriptome data. Furthermore, SORE may also be able to enhance RRH-mediated light perception (+54%), as its expression was higher in SORE-treated, light-exposed skin explants than in placebo-treated, light-exposed skin explants.

[0174] [Table 9]

[0175] Example 8: SORE regulates the circadian rhythm of skin cells Results were obtained from human skin explant experiments described in Example 5.

[0176] [Table 10]

[0177] - TOP2A plays a role in regulating the period length of ARNTL / BMAL1 transcriptional oscillations, affecting the circadian rhythm.

[0178] - SUV39H2 can be recruited by the large PER complex to E-box elements of circadian target genes such as PER2 itself or PER1.

[0179] Both SFPQ and PSPC1 may regulate the circadian clock by suppressing the transcriptional activator activity of the CLOCK-ARNTL / BMAL1 heterodimer, which is required for transcriptional repression of circadian target genes, such as PER1, mediated by the large PER complex via histone deacetylation.

[0180] - TIMELESS is involved in period length determination and DNA damage-dependent phase progression of the circadian clock.Timeless negatively regulates CLOCK|NPAS2-ARTNL / BMAL1|ARTNL2 / BMAL2-induced transactivation of PER1, possibly via nuclear translocation of PER1.

[0181] - HNRNPU has many functions, including involvement in the circadian regulation of the core clock component ARNTL / BMAL1 transcription.

[0182] - PASD1 functions as a suppressor of the circadian clock that drives the daily circadian rhythms of cells throughout the body. It can act as a nuclear repressor of CLOCK-ARNTL / BMAL1 heterodimer-mediated transcriptional activation of core clock components.

[0183] All these results demonstrate that SORE can regulate the circadian rhythm of skin cells, which underlies the regulation of a wide range of cellular, metabolic, physiological and behavioral activities in mammals.

[0184] Example 9: SORE maintains cellular integrity by inducing DNA repair mechanisms The results were obtained from human skin explant experiments described in Example 5. SORE showed cytoprotective effects by upregulating DNA repair players (Table 11).

[0185] [Table 11]

[0186] - MBD4 is a mismatch-specific DNA N-glycosylase involved in DNA repair. MBD4 has thymine glycosylase activity and is specific for G:T mismatches within methylated and unmethylated CpG sites. It also removes uracil or 5-fluorouracil from G:U mismatches.

[0187] - TREX1 is the primary cellular 3'→5' DNA exonuclease that can digest single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) with mismatched 3' ends.

[0188] - BOD1L1 is a component of the fork protection mechanism required to protect stalled / damaged replication forks from uncontrolled DNA2-dependent resection. It acts by stabilizing RAD51 at stalled replication forks and protecting RAD51 nucleofilaments from antirecombinogenic activity.

[0189] - RAD51B participates in the homologous recombination repair (HRR) pathway of double-stranded DNA breaks that occur during DNA replication or are induced by DNA damaging agents. It promotes the assembly of presynaptic RAD51 nucleoprotein filaments.

[0190] - KIN is involved in DNA replication and the cellular response to DNA damage. It participates in DNA replication factories and creates a bridge between DNA replication and repair mediated by high molecular weight complexes.

[0191] - MSH2 is a component of the post-replicative DNA mismatch repair system (MMR), which binds to DNA mismatches and initiates DNA repair.

[0192] - FANCD2 is required for maintaining chromosomal stability and is involved in the repair of DNA double-strand breaks by both homologous recombination and single-strand annealing.

[0193] - RAP3 is part of the heterotrimeric replication protein A complex (RPA / RP-A) and binds to and stabilizes single-stranded DNA intermediates that form during DNA replication or upon DNA stress, thereby playing an essential role in both DNA replication and the cellular response to DNA damage.

[0194] - POLI is an error-prone DNA polymerase specifically involved in DNA repair.

[0195] Several upregulated systems of DNA repair are highly specific to environmental damage, particularly from UV light.

[0196] XPA is involved in DNA excision repair. It initiates repair by binding to the damaged site with varying affinities, depending on the photoproduct and transcriptional state of the region. It is also required for UV-induced CHEK1 phosphorylation and recruitment of CEP164 to the cyclobutane pyrimidine dimer (CPD) of DNA damage after UV irradiation.

[0197] - SLF1 plays a role in the DNA damage response (DDR) pathway by regulating post-replicative repair of UV-damaged DNA and maintaining genome stability. Complementarily, NSMCE4A (non-structural maintenance of chromosomes element 4 homolog A) is a component of the SMC5-SMC6 complex, which is involved in DNA double-strand break repair via homologous recombination.

[0198] - ATR activates checkpoint signaling upon genotoxic stress, such as ionizing radiation (IR), ultraviolet light (UV), or DNA replication arrest, thereby acting as a DNA damage sensor.

[0199] - PRIMPOL is a DNA primase and DNA polymerase that is required to tolerate replication-terminating lesions by bypassing them. It provides various translesion synthesis options when DNA replication is halted. It synthesizes DNA primers downstream of lesions such as ultraviolet (UV) lesions, R-loops, and G-quadruplexes, allowing DNA replication to continue.

[0200] Thus, SORE acts on several DNA repair components involved in normal or environmentally induced DNA damage, and it may be a powerful protector in protecting DNA integrity, which can be compromised by both the aging process and / or environmental aggressors such as sunlight.

[0201] Example 10: Anti-collagenase inhibitory activity of FDTA-I and FDTA-II purified from SORE FDTA-I and FDTA-II were purified from stimulated roots of Sanguisorba officinalis. Plants were stimulated for 1–3 weeks using a nitrogen-free nutrient medium (N / P / K 0 / 15 / 40). 10 g of dried and ground (1 mm) roots were macerated in 100 ml of ethyl acetate at 50°C for 24 hours. Purified fractions of FDTA-I and FDTA-II were obtained by preparative HPLC and characterized by mass spectrometry. Ferulic acid and TA were purchased commercially. Additionally, diglycoside I, an active compound isolated from Sanguisorba officinalis roots known to possess various activities, including increasing the expression of type I collagen, was also included in the study. For comparison, SORE was obtained as shown in Example 1. Anticollagenase inhibitory activity was measured by the kinetics of collagenase-driven degradation of the following collagenase substrate: FALGPA (N-[3-(2-furyl)acryloyl]-Leu-Gly-Pro-Ala) to FAL (N-[3-(2-furyl)acryloyl]-Leu) and Gly-Pro-Ala).

[0202] Briefly, a 100 μL volume of a 1.5 mM solution of FALGPA in 50 mM Tricine buffer (Bachem, 4006713.0025) containing 10 mM calcium chloride and 400 mM sodium chloride was mixed with a 10 μL volume of test sample or the test sample's pure solvent (positive control), followed by 10 μL of 0.05 mg / mL Clostridium histolyticum collagenase (type IA, Sigma-Aldrich, C9891). To determine the rate of enzymatic conversion, the content of FAL was quantified by stopping the enzymatic conversion of FALGPA with EDTA at the start of the reaction and 30 min after its initiation. To quench the enzymatic conversion, a 50 μL volume of the test sample was mixed with a 50 μL volume of EDTA (0.2 M in water). The FAL content in the samples was then quantified using an UHPLC method optimized for the separation of FALGPA and FAL. Analysis was performed using a Kinetex biphenyl reversed-phase column (150 mm x 2.1 mm, 2.6 μm, Phenomenex, Torrance, USA) maintained at 40 °C. The mobile phase consisted of water containing 0.1% by volume formic acid (A) and pure acetonitrile (B), delivered at 0.5 mL / min with the following B-phase gradient: 5–41% (0–9 min); 41–90% (9–9.05 min); hold at 90% (9.05–11.50 min); 90–5% (11.50–11.55 min); hold at 5% (11.55–14.50 min). The sample injection volume was 5 μL, and FALGPA / FAL detection was at 338 nm. The residual activity and degree of inhibition of collagenase in the presence of test sample X was then calculated using the following formula:

[0203]

number

[0204] [Table 12]

[0205] SOREs enriched with FDTA-I and FDTA-II inhibited collagenase IA activity (50%) at very low concentrations (0.04%). FDTA-I and FDTA-II acted as collagenase IA inhibitors at 25 μM (76% and 36% inhibition, respectively), whereas neither TA nor ferulic acid alone had any effect on the enzyme at the same concentration. This clearly demonstrated the relevance of the coupling of ferulic acid to TA via an ester bond in the context of its anti-collagenase bioactivity. Interestingly, diglycoside I did not have any enzyme inhibitory effect at 25 μM.

[0206] These results demonstrate the ability of SOREs, including FDTA-I and FDTA-II, to inhibit collagenase activity. Therefore, SOREs can be used to improve the mechanical properties of skin, particularly ECM components, by inhibiting collagenase activity. Furthermore, their activity against collagenase is apparently independent of the activity reported for diglycoside I.

[0207] Example 11: Anti-hyaluronidase inhibitory activity of FDTA-I and FDTA-II purified from SORE FDTA-I and FDTA-II were purified from stimulated roots of Sanguisorba officinalis as described in Example 10. Additionally, diglycoside I was also included in the study. For comparison, SORE was prepared as shown in Example 1.

[0208] Hyaluronidase catalyzes the degradation of hyaluronic acid (HA). The assay is based on quantifying the intact HA content in the presence / absence of an extract and / or compound of interest. The rate of degradation of the HA content is proportional to the enzymatic activity of hyaluronidase and inversely proportional to the inhibitory power of the extract.

[0209] The reaction mixture was prepared as follows: 37.5 μL of 20 mM PHB buffer (pH 7), 187.5 μL of hyaluronidase solution (bovine hyaluronidase, type IS, Sigma) 0.02 mg / mL, and 25 μL of sample (SORE or pure FDTA-I and FDTA-II to be tested). In Table 13, pure FDTA-I and FDTA-II and SORE were in pure propane-1,3-diol. In Table 14, pure FDTA-I and FDTA-II were in 100% ethanol. The mixture was incubated at room temperature for 10 minutes, and then 250 μL of HA substrate (0.3 mg / mL in phosphate buffer) was added to initiate the reaction. The incubation was carried out in a Thermomixer at 37°C with stirring (450 rpm). The reaction mixture (50 μL) was sampled every 15 minutes to monitor the reaction for 60 minutes. These samples were inserted into wells of a microplate containing 200 μL of 0.1% BSA solution (pH 3.75) and incubated in BSA for 10 minutes. HA formed aggregates with the BSA solution, which precipitated. The amount of precipitated BSA was therefore proportional to the optical density at 600 nm, which allowed the enzyme reaction to be monitored. The microplate was finally inserted into a spectrophotometer, and the optical density was measured at this wavelength.

[0210] [Table 13]

[0211] FDTA-I acted as a hyaluronidase inhibitor at 15 μM, whereas neither TA nor ferulic acid alone had any effect on the enzyme at the same concentration, clearly demonstrating the relevance of the coupling of ferulic acid to TA via an ester bond in the context of its antihyaluronidase bioactivity.

[0212] [Table 14]

[0213] FDTA-I and FDTA-II acted as hyaluronidase inhibitors at 15 μM, whereas diglycoside I had no effect on this enzyme at the same concentration. These results demonstrated the ability of SOREs containing FDTA-I and FDTA-II to inhibit hyaluronidase activity. Therefore, SOREs can be used to improve the mechanical properties of skin, particularly ECM components, by inhibiting hyaluronidase activity.

[0214] Example 12: SORE improves skin elasticity and skin tone Another clinical study conducted on Caucasian women showed that the calculation of the Ue parameter allows the determination of the elastic properties of the skin, which are well-documented to decrease with age. (登録商標) An improvement in skin elasticity was demonstrated as measured using a cutometer (Courage & Khazaka).

[0215] Figure 6 shows that SORE improves skin elasticity as evidenced by an increase in the Ue parameter after 28 days of use (+15% *: p<0.05), while placebo had no significant effect (+5% ns), which was consistent with the data obtained in vitro and described in Examples 10 and 11, which show that FTDA-I and FTDA-II inhibit collagenase and hyaluronidase activity, which are known to alter mechanical properties of the skin, such as elasticity and firmness.

[0216] Clinical Study 2 also confirmed that SORE improved skin tone after 28 days of application. Facial tone uniformity was scored on an unstructured scale using Visio-Face with a high-resolution Nikon D300 camera. (登録商標) Photographs were taken at n°2. Finally, the volunteers' self-evaluation of the cream was assessed through a questionnaire. Figure 7 shows that SORE improved the complexion of the skin, which appeared more homogeneous, with a +16% improvement in the homogeneity score (*: p<0.05), whereas placebo did not (+4% ns).

[0217] This beneficial effect was confirmed by the volunteers themselves, as over 73% of the volunteers perceived their complexion to be more even, as opposed to placebo (52%), after 28 days of topical application of SORE (data not shown).

[0218] Example 13: SORE reduces protein carbonylation in keratinocytes Since it has been recognized that increased protein carbonylation in the skin is associated with changes in skin quality, such as skin tone or fragile dermis, it was decided to also analyze the effect of SORE on protein carbonylation in keratinocytes in the clinical trial described in Example 5.

[0219] Surface keratinocytes were collected from volunteers' skin before (day 0) and after 28 days of treatment with 1% SORE (see Table 9) or placebo. D-squames adhesive was applied for 5–20 seconds using a specific applicator (stable pressure: 50 g). The first sample was not retained; only the second sample was considered. The strips were then immersed in a dissociation solution containing 20% ​​SDS, 20 mM DL-dithiothreitol, 5 mM EDTA, and 0.1 M Tris-HCl buffer, pH 8, at 100°C for 10 minutes. The tubes were then centrifuged. After three 1-minute washes in the dissociation solution at room temperature and recentrifuged, the keratinocytes were labeled with 2 mM fluorescein-5-thiosemicarbazide in 0.1 M 2-morpholinoethanesulfonic acid (MES)-Na buffer, pH 5.5, for 1 hour at room temperature. After three washes in phosphate-buffered saline (PBS), the suspension was placed on a microscope slide for 12 hours. Five photographs were taken per slide. Fluorescence intensity was quantified using ImageJ software. Results showed that carbonylated protein content was significantly reduced in the keratinocytes of volunteers treated with SORE compared to placebo after 28 days of application (Figure 8). These results demonstrate the ability of SORE to protect skin from the harmful effects of carbonylated proteins and thus improve skin quality, including skin tone.

Claims

1. below: tormentic acid, representing at least 1% by weight relative to the total weight of the dry extract; feruloylated derivatives of tormentic acid having the general formula (I) 【Chemistry 1】 - feruloylated derivatives of deoxytormentic acid, having the general formula (II) 【Chemistry 2】 2. A root extract of Sanguisorba officinalis, comprising:

2. 2. The root extract of claim 1, wherein the feruloylated derivative of deoxytormentic acid represents at least 0.05% by weight relative to the total weight of the dry extract.

3. 3. A root extract according to claim 1 or 2, wherein the feruloylated derivative of tormentic acid represents at least 0.1% by weight relative to the total weight of the dry extract.

4. 4. The root extract according to claim 1, wherein the tormentic acid, the feruloylated derivative of tormentic acid and the feruloylated derivative of deoxytormentic acid together represent at least 1.15% by weight, relative to the total weight of the dry extract.

5. 5. The root extract of any one of claims 1 to 4, further comprising one or more ellagitannins.

6. Steps below: a) cultivating Sanguisorba officinalis under soil-free conditions, in particular aeroponic; b) stimulating the roots of the plant; c) solid / liquid extraction by maceration of the roots obtained in step b); d) recovering the extract obtained in step c), and e) optionally diluting and / or clarifying the extract recovered in step d) by successive filtration; 6. A method for producing the root extract of the plant Sanguisorba officinalis according to any one of claims 1 to 5, comprising:

7. below: - a Sanguisorba officinalis root extract according to any one of claims 1 to 5 or a Sanguisorba officinalis root obtained by the method according to claim 6; and at least one cosmetically or dermatologically acceptable ingredient other than an extract of the root of Sanguisorba officinalis, 1. A cosmetic or dermatological composition comprising: a composition for topical use selected from the group consisting of a solution, suspension, emulsion, cream, paste, gel, lotion, powder, soap, surfactant-containing water, oil, shampoo, and spray; or a nutraceutical composition for oral administration.

8. Cosmetic use of a Sanguisorba officinalis root extract according to any one of claims 1 to 5 as an active ingredient for promoting light-related human mood, for skin care and / or scalp care.

9. 9. Cosmetic use according to claim 8 for increasing and / or stimulating the production of at least one well-being hormone in skin cells.

10. 10. Cosmetic use according to claim 8 or 9 for promoting the production of at least one happiness hormone in skin cells.

11. Cosmetic use according to any one of claims 8 to 10 for promoting or stimulating the expression of phototransduction molecules, preferably peropsin (RRH), in skin cells.

12. Cosmetic use according to any one of claims 8 to 11 for maintaining or restoring the circadian rhythm of clock proteins in skin cells.

13. Cosmetic use according to any one of claims 8 to 12 for stimulating vitamin D downstream biological pathways.

14. 14. The cosmetic use according to any one of claims 8 or 13 for preventing or delaying the signs of photoaging in the skin, for preventing or treating UV-related skin damage, or a combination of two or more thereof.

15. 15. Cosmetic use according to claim 14 for reducing and / or preventing protein carbonylation in skin cells.

16. 16. Cosmetic use according to claim 14 or 15 for improving the mechanical properties of the skin.

17. 17. The cosmetic use according to any one of claims 14 to 16 for improving skin elasticity, improving skin complexion, increasing dermal density, maintaining or restoring skin cell integrity, maintaining or restoring skin barrier function, promoting skin hydration, improving skin tone, preventing the formation of fine lines and wrinkles, or a combination of two or more thereof.

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