OA21067A - Moringa Peregrina seed extract rich in 2,5-diformylfuran, its process for obtaining it and its use in cosmetic compositions. - Google Patents

Abstract

The invention relates to an extract of Moringa peregrina seeds rich in the compound 2,5diformylfuran, more specifically an extract of the cake of said seeds and a process for extracting the extract. It also relates to cosmetic or nutricosmetic compositions comprising said extract and the use of said compositions to improve the appearance of the skin, mucous membranes or appendages, to relax, soothe and de-stress the skin and prevent and/or fight against skin signs of aging and/or photo-aging and prevent age spots.

Description

Description

Title of the invention: Moringa peregrina seed extract rich in 2,5-diformylfuran, its process for obtaining it and its use in cosmetic compositions

Technical area

[0001] The invention relates to the field of cosmetics and nutricosmetics and more particularly that of active ingredients used in the formulation of compositions for skin care. The invention relates to an extract of Moringa peregrina seeds rich in the compound 2,5-diiomiylfuran (DFF). The invention also relates to the process for obtaining a particular extract from the seeds of Moringa peregrina, the cosmetic compositions comprising such extracts, and finally to the cosmetic or nutricosmetic use of such compositions for the care of the skin, scalp and dander.

Technology background

[0002] The Moringaceae constitute a mono-generic family (a single genus, Moringa Adans), element of the Indian Saharan flora, made up of between twelve and fourteen species according to the authors, distributed from eastern Africa to Asia. The genus is classically divided into 3 sections which are, however, not confirmed as monophyletic by phylogenetic analyses. The latter rather highlighted clades focused on certain morphological characters: pachycaules (“boule trees”); tuberous trees and neither pachycaules nor tuberous trees (in English “slender trees”). The species Moringa peregrina (Forssk.) Fiori belongs to the third group. The few genetic studies on the genus or family confirm the reality of the species compared to other species in the genus, particularly with regard to the Indian Moringa, Moringa oleifera Lam. (see in particular the articles: OLSON Μ. E. 2002, Combining Data from DNA Sequences and Morphology for a Phylogeny of Moringaceae (Brassicales), Systematic Botany 27(1): 55-73; Hassanejn A. M. A. and ALSOQEE, A. A., 2018, Morphological and genetic diversity of Moringa oleifera and Moringa peregrina genotypes, Horticulture, Environment and Biotechnology 59(2): 251-261). A recent article on Moringa peregrina sampled from different localities in Saudi Arabia concluded, using ITS markers, that the species is genetically stable (ALAKLABJ A., 2015, Genetic diversity of Moringa peregrina species in Saudi Arabia with ITS sequences , Saudi Journal of Biological Sciences 22: 186-190) with, however, a high level of intra-population genetic variation.

[0003] The species Moringa peregrina is found in the rocky environments of Yemen, Oman, and Saudi Arabia, in East Africa, in Sudan, in Ethiopia,

Eritrea, Somalia and Djibouti. Its presence in Iran seems limited to the southern provinces but requires confirmation (PROTA14 = Munyanziza E. ET Yongabi K. A..Vegetable oils/Oléagineitx, Moringa peregrina (Forssk.) Fîori, http://database.prola.org/piOtahlml/moringa peregrina_fr.htm, accessed 10/23/2019). In the Levant and Egypt, the species today only represents rare, scattered, relict stations (with the exception of a few altitudinal populations), mainly in the Sudanian region. Moringa peregrina is also considered rare and endangered today in Sudan and Yemen. Moringa peregrina occupies the driest and most inhospitable habitats compared to the other species of its clade. It is apparently more drought tolerant than Moringa oleifera which is planted commercially on a large scale in tropical and subtropical areas. Recent work has shown that the size and size of seeds has a positive impact on germination time and the rate and speed of growth of young individuals (Gomaa N. H. ET P1CÔ F. X., 2011, Seed germination, seedling traits, and seed bank of the tree Moringa peregrina (Moringaceae) in a hyper-arid environment, American Journal of Botany 98(6): 1024-1030), indicating an adjustment in resource allocation on seed quality rather than number, which allows for Moringa peregrina to reproduce effectively in extreme abiotic (hyperarid) environments. Moringa peregrina seeds have a thicker central mesotesta, in terms of cell layer, than those of M oringaoleife ra.

There are some historical records which tend to indicate that Moringa peregrina oil was actively traded in the early days of Islam in the AlUla region (NASEEF, AAS, 1995, Al-'Vlâ, A study of Cultural and Social Heritage). The oil produced locally from Moringa peregrina is today mainly intended for self-consumption or local markets. In Saudi Arabia, the leaves were traditionally used as a decoction for internal use to treat diabetes, colon diseases, eye diseases and anemia (Abdel-Kader MS, Hazazi AMA, Elmakki OA and AlqasOUMI SI, 2018, A survey on the traditional plants used in Al Kobah village, Saudi Pharmaceutical Journal 26(6): 817-821) and as a diuretic, rubefacient and astringent (AQEEL AAM, TAR1Q Μ., M OSSA JS, Al-Yahya MA and Al-Saud MS , 1984, “Plants used in Arabia Folk medicine”, Report submitted to Saudi Arabian National Center for Science and Technology, Riyadh, Saudi Arabia). In Oman, the oil, extracted by women at the end of summer, is used against migraines, fever, burns, lacerations and fractures, constipation and stomach pain, against muscle pain and against hair dryness, against childbirth pains (GHAZANFAR SA, 1994, Handbook of Arabian Medicinal Plants, l ^sl ed., CRC Press, Boca Raton, Ann

Harbor, s.u. ; Ghazanfar S.A., 1998, Plants of Economic Importance, cap. 15, in G HAZANFAR, S.A. AND F1SHER, M. (ed.) Vegetation of the Arabian Peninsula. Geobotany 25, p. 241-264, Kluwer Academie Publishers, table 11.1, p. 247 and 11.7 p. 251). It was also used in perfume compositions (Ghazanfar S.A., 1998, p. 259) and in Oman and Yemen as a facial lotion (Ghazanfar S.A. AND RECHINGER B., 1996, Two multi-purpose seed oils form Oman. Plants for Food and Medicine. Paper presented at the joint meeting of the Society for Economic Botany and International Society for Ethnopharmacology, 1-7 July 1996, London).

[0004] Extracts from the seed of Moringa oleifera are known in the cosmetic field. For example, we know from document FR296879 an extract of whole seeds (with integuments) of Moringa oleifera containing oil (including triglycerides, fatty acids and polar lipids) and polyphenols and its use in cosmetic compositions to fight against skin aging . In this document, it is the non-polar part of the Moringa oleifera seed which appears to be active, and more particularly the oily part. We also know from document FR2776519 that protein extracts of Moringa oleifera seeds, known for their clarifying effects on turbid waters, have a softening, physiological conditioning, hydrating, restructuring, restorative and as a softening effect on the skin and mucous membranes. anti-pollution active ingredient. In this document the active ingredients are proteins with molecular weights between 6,500 and 8,800 Da which are obtained by aqueous extraction on Moringa oleifera cake. We also know the document FR3076460 which concerns the use of a protein extract of ungerminated and deoiled seed of Moringa oleifera for the treatment of sensitive, sensitized, reactive, fragile and/or weakened skin and/or mucous membranes and/or in the treatment and /or prevention of erythema, particularly diaper rash in infants. In this document, the extraction process makes it possible to obtain a major fraction of proteins whose molecular weights are approximately 8800 Da. We also know from document KR20130088224 the use of an extract of whole germinated seed of Moringa oleifera in cosmetics, in particular obtained by extraction using a supercritical fluid. This process makes it possible to isolate non-polar carotenoids and amino acids which are described as active for whitening cosmetic use. All the aforementioned documents concern the use of the species Moringa oleifera. none describes the use in the cosmetic field of the Moringa peregrina species. In the document by Kolheil et al., XP055753955, 2011, the extraction with ethanol of bioactive polyphenolic compounds, tannins, flavonoids, saponins, unsaturated sterols and/or whole seeds of Moringa peregrina is disclosed. or triterpenes. The extract obtained is stored at 4°C and has an anti-oxidant effect. In the document by Abbas Alba et al., XP055753970, 2018, an ethanolic extraction for three days or more is disclosed on whole seeds of Moringa peregrina. The concentration of the filtrates is obtained under reduced pressure at a temperature between 45 and 50°C. In the document by Abou-Hashcm et al., XP055754018, 2019, an ethanolic extraction for three times 72 hours is disclosed on whole Moringa peregrina seeds. The extract obtained is then filtered and concentrated with a rotary evaporator at a temperature of approximately 45°C. In the document by Majali Ibrahim el al., XP055754048, 2015, an extraction with ethanol with stirring for half an hour and then precipitation for 72 hours is disclosed on whole seeds of Moringa peregrina. None of the aforementioned documents discloses the ethanolic extraction on the meal of the unhulled seed of Moringa peregrina.

[0005] More specifically for the species Moringa peregrina, we know that certain phenolic compounds and flavonoids obtained from leaves or whole seeds of Moringa peregrina exhibit antioxidant activity (Al-Dabbas M., 2017, Antioxidant activity of different extracts from the aerial part of Moringa peregrina (Forssk.) Fiori, from Jordan, Pakistan Journal of Pharmaceutical Sciences, 30(6): 2151-2157). These compounds are extracted with solvents such as methanol, ethyl acetate or hexane from leaves or whole seeds. It appears that it is the leaves which contain the most active compounds.

[0006] Thus, depending on the species used in the Moringa genus, it is observed that depending on the part of the plants (leaf, seed), the part of the seeds (whole or not, shelled or not) and the extraction process used implemented, in particular the choice of solvent, the extracted molecules turn out to be different. However, the composition of the extract determines the biological activity and consequently the cosmetic effectiveness.

Considering the above, a problem that the invention proposes to solve is to develop new products based on an extract of the species M. peregrina of the genus Moringa which can be used in cosmetics and are easy to use. artwork.

[0008] As a result, the Applicant has demonstrated a new extract obtained from the seeds and more specifically from the seed cake of the species Moringa peregrina, which presents in particular a relaxing and anti-stress activity for the skin, a anti-aging activity as well as preventive activity for age spots. The extract according to the invention is rich in 2,5-diformylfuran (DFF), a compound also called 2,5-furandicarboxaldehyde. The extract is obtained specifically from the seeds or more specifically from the unshelled seed cake of Moringa peregrina, in particular by alcoholic extraction. The extract according to the invention is new in two ways in the field of cosmetics compared to the extracts of the prior art, on the one hand by the specific species of origin used and on the other hand by its content in particular compounds.

[0009] By intergovernmental agreement of April 10, 2018 between the government of the French Public Republic and the Kingdom of Saudi Arabia, the applicant, Agence Française Pour Le Développement d'AIUla (AFALULA) as well as the Royal Commission for A1U1A (RCU) have a common project in particular to develop responsible agriculture and the local economy, in particular through the local production of natural products derived from indigenous plants and to protect the biodiversity and rights of the AIUla region of the Kingdom of Saudi Arabia. The Kingdom of Saudi Arabia has been a member of the Nagoya Protocol since October 8, 2020. At the time of writing this patent, the implementing regulations under which the Nagoya Protocol will be incorporated into relevant aspects of local law are in force. exam course. Therefore, at this stage, the Kingdom of Saudi Arabia has no specific requirements with regard to this patent application and the Nagoya Protocol. Thus, on the day the patent application is filed, there is no requirement for a certificate of conformity regarding access to genetic resources.

Summary

[0010] The first subject of the invention is an extract of Moringa peregrina seeds rich in the compound 2,5-diformylfuran. The compound 2,5-diformylfuran is a rare saccharide compound in the plant kingdom and synthesized from furfural via the intermediate synthesis of 5 hydroxymethylfurfural.

[0011] Thanks to its characteristic, a concentration rich in 2,5-diformylfuran, the extract according to the invention is unique in the Moringa genus. It will be demonstrated that the species Moringa peregrina presents a particular molecular profile different from those known from other species of the genus, in particular from the species Moringa oleifera, which the Applicant has been able to demonstrate.

[0012] The second object of the invention is a process for obtaining an extract of Moringa peregrina seeds according to the invention, comprising the following steps according to which:

a) the unshelled Moringaperegrina seeds are collected and dried to obtain an internal humidity level of less than 8%;

b) the dried seeds are pressed so as to separate the oil from the rest of the seed to obtain the cake,

c) we grind the cake obtained in step b),

d) the ground material obtained in step c) is dispersed in a proportion of approximately 25% by weight of solid material relative to the total weight used in a predominantly alcoholic solvent, the alcohol being chosen from ethanol or methanol with optionally a co-solvent of the polyol or subcritical water type, in a proportion of 80 to 100% by weight of alcohol relative to the total weight of the solvent;

e) a solid-liquid extraction is carried out, with stirring, at a temperature between 16 and 30°C for a period of approximately 2 hours,

f) the liquid and solid phases are separated so as to eliminate the solid phase and recover a liquid extract of Moringa peregrina cake, and

g) optionally, the liquid Moringa peregrina extract obtained is dried so as to obtain a solid Moringa peregrina extract.

[0013] The third object of the invention is a cosmetic or nutricosmetic composition comprising, as active agent, an effective quantity of a Moringa peregrina seed extract according to the invention and a physiologically acceptable excipient.

[0014] Finally, the fourth object of the invention is the cosmetic or nutricosmetic use of a composition according to the invention, to improve the appearance of the skin, mucous membranes or appendages, to relax, soothe and de-stress the skin and prevent and/or fight against the skin signs of aging and/or photo-aging, to prevent age spots.

Brief description of the figures

[0015] The invention will be better understood, and other aims, details, characteristics and advantages thereof will appear more clearly during the following description of several particular embodiments of the invention, given solely by way of illustration. and not limiting. Description of embodiments

[0016] In this description, unless otherwise specified, it is understood that, when an interval is given, it includes the upper and lower limits of said interval.

[0017] In the present invention, the following abbreviations mean:

- MTT: 3-(4,5-dimethylthiazol-2-yI)-2,5-diphenyl tetrazolium bromide (MTT test is a rapid method for counting living cells)

- SDS: Sodium Dodecyl Sulfate

- PBS: Phosphate Buffer Saline

- ELISA: Enzyme-Linked Immunosorbent Assay

- PCR: Polymerase Chain Reaction

- ANOVA: Analysis Of Variance

- MSH: Melanocyte Stimulating Hormone

[0018] In the present invention, we mean:

- “extract rich in 2,5-diformylfuran compound” an extract containing a quantity of 2,5-diformylfuran compound greater than the other identified ingredients, i.e. a quantity greater than 50% relative to the dry matter of the total extract.

- “effective quantity” the necessary quantity of active molecules to obtain the desired result, namely, to make it possible in particular to obtain protection of the extra-cellular matrix of the skin.

- “topical application”, the act of applying or spreading the active principle according to the invention, or a composition containing it, to the surface of the skin, a mucous membrane or the integuments.

- “physiologically acceptable” which is suitable for topical use, in contact with human skin, or for use by other routes of administration, for example the oral route or injection into the skin, without risk of toxicity, incompatibility, instability, allergic response.

- “cake”, the deoiled part of the seed after pressing. This is the solid residue from the extraction of oil from the seeds. It is a co-product of crushing, the oil manufacturing process. It generally represents 50 to 75% of the mass of the seeds. - “unshelled seeds”, the fact that the shell, or pericarp, of the harvested seeds is maintained around the seeds.

- “when the fruit is ripe”, the fact that the fruit is ripe, preferably when the pod is at the beginning of dehiscence and it takes on a dark beige to brown color and when a 180° twist is applied to the lower quarter of the fruit. the pod causes the valves to begin to open.

- “predominantly alcoholic solvent”, the fact that the alcoholic type solvent can include a co-solvent having sufficient characteristics to extract the active principle, knowing that pure 96° ethanol appears to be the most suitable alcoholic solvent.

- “approximately” a margin more or less of 10% to 20% in relation to the information given (duration, percentage, etc.).

- “active molecule”, also “active principle”, the molecule of 2,5-diformylfuran extracted according to the process of the invention from the seeds of Moringa peregrina. This molecule is responsible for the biological activities described in the present invention.

- “active agent” a sufficient quantity of an extract according to the invention to obtain the biological activities described. Depending on whether the extract is liquid or dried, concentrated or not, the quantities of the active agent can vary in proportions from 0.002 to 20% by weight relative to the total weight of the composition.

- “signs of skin aging”, all changes in the external appearance of the skin and appendages due to aging such as, for example, wrinkles and fine lines, withered skin, flabby skin, thinning skin, lack of elasticity and/or tone of the skin, dull and lackluster skin or pigmentation spots on the skin, discoloration of the hair or stains on the nails, but also any intentional change in the skin that does not appear systematically by a modified external appearance such as, for example, any internal degradation of the skin following exposure to ultraviolet (UV) radiation.

The invention primarily relates to an extract of Moringa peregrina seeds rich in the compound 2,5-diformylfuran. The molecule 2,5-diformylfuran, also called 2,5-furandicarboxaldehyde, has never been characterized in an extract from the seeds of Moringa species. The Moringa peregrina species grows in very arid climates. Thus, its ability to resist drought allowed it to acquire particular unique characteristics, which the Applicant was able to identify by implementing an extraction process adapted to whole seeds or preferably to seed cake. .

[0020] In the context of the present invention, the plant part chosen is the seed of Moringa peregrina. It is known that Moringa peregrina seeds are used for the extraction of their oil called peregrina oil (INCI name “Moringa peregrina seed oil”), used regionally for self-consumption or in various traditional medicinal indications. The cake obtained after the seed has been deoiled is a waste currently used for animal feed in particular.

[0021] According to the first object of the invention, the extract of Moringa peregrina is obtained by solid-liquid extraction of the cake from the unshelled seeds, with stirring, in a proportion of approximately 25% by weight of solid material relative to by total weight used in a predominantly alcoholic solvent, the alcohol being chosen from ethanol or methanol with optionally a co-solvent of polyol or subcritical water type, in a proportion of 70 to 100% by weight of alcohol relative to the total weight of the solvent, at a temperature between 16 and 30°C for a period of approximately 2 hours, and by separation of the liquid and solid phases so as to eliminate the solid phase and recover a liquid extract of seed of Moringa peregrina, said extract being rich in the compound 2,5-diformylfuran. Preferably, the extract according to the invention is obtained from the cake of seeds harvested at maturity from the fruit of Moringa peregrina after extraction of the peregrina oil (INCI name “Moringa peregrina seed oil).

[0022] It should be noted that the active principle of the extract according to the invention, namely 2,5-diformylfuran, is a molecule of mixed polarity presenting a certain fragility. Thus, the extract obtained from the whole, unshelled seed must be selectively extracted to obtain a high concentration of active ingredients, using appropriate solvents and co-solvents and a temperature not exceeding 30°. vs.

The co-solvents can be for example glycol ethers (mono- or dipropylene glycol, propanediol, and other propylene glycol derivatives, ethylene or diethylene glycol derivatives), glycerol, dimethyl ether isosorbide , methyl or ethyl or propyl esters of fatty acids; dicraprylyl carbonate, dicaprylyl ether, alkyl acetate or proprionate, acetone, methyl or ethyl ketone, monoterpenes such as alpha-pinene or limonene, The proportions of these co-solvents can be mixed with the primary solvent (e.g. ethanol or methanol ) from 0 to 30% (V/V).

The extraction conditions can be under atmospheric pressure or under vacuum or under an inert atmosphere, but preferably in the dark at a temperature between 16 and 30°C.

[0025] In a preferred embodiment according to the invention, the extract is obtained from the cake of unshelled seeds by solid-liquid extraction, with a 96° ethanol alcoholic solvent.

[0026] In yet another embodiment, the liquid extract obtained is dried so as to obtain a dry extract of the Moringa peregrina seed cake containing more than 50% by weight of 2,5-diformylfuran relative to the total weight. dry matter,

[0027] The dry extract of Moringa peregrina seed cake contains more precisely approximately 55% by weight of 2,5-diformylfuran, 2.5% furfural, 1.2% isopropyl myristate, 4.7% palmitic acid, 11.1 % oleic acid and 25.8% triglycerides relative to the total weight of dry matter.

[0028] The second object of the invention is a process for obtaining an extract of Moringa peregrina seeds according to the invention comprising the following steps according to which:

a) the unshelled seeds of Moringa peregrina are collected and dried to obtain an internal humidity level of less than 8%;

b) the dried seeds are pressed so as to separate the oil from the rest of the seed to obtain the cake,

c) we grind the cake obtained in step b),

d) the ground material obtained in step c) is dispersed in a proportion of approximately 25% by weight of solid material relative to the total weight used in a predominantly alcoholic solvent, the alcohol being chosen from ethanol or methanol with optionally a co-solvent of the polyol or subcritical water type, in a proportion of 70 to 100% by weight of alcohol relative to the total weight of the solvent;

e) a solid-liquid extraction is carried out, with stirring, at a temperature between 16 and 30°C for a period of approximately 2 hours,

f) the liquid and solid phases are separated so as to eliminate the solid phase and recover a liquid extract of Moringa peregrina cake, and

g) optionally, when the alcohol is ethanol, the liquid Moringa peregrina extract obtained is dried so as to obtain a solid Moringa peregrina extract.

[0029] In a preferred embodiment, the unshelled seeds are collected, that is to say the shell of which is kept, when the fruit is ripe and preferably when the pod is at the beginning of dehiscence.

[0030] In a preferred embodiment, the seeds are dried to obtain an internal humidity level of around 6%, drying will preferably be done on a ventilated shelf sheltered from the sun's rays, preferably under a shade in the open air. .

[0031] The dried seeds are then crushed extemporaneously with cold pressing which makes it possible to mechanically separate the peregrina oil (INCI name “Moringa peregrina seed oil”) from the rest of the compressed seed, namely the cake.

[0032] The cake is then crushed mechanically with all types of mechanical crushers such as hammer, flail, knife, crushing/shredding systems.

The extraction is advantageously always carried out with stirring, thus allowing dispersion and homogenization of the solid in the liquid, improving the diffusion of the solute in the solvent.

To mainly extract the 2,5-diformylfuran compound of interest, an alcoholic solvent of the 96° ethanol type will be preferred, but methanol can also be used, with a co-solvent of the polyol type or water. subcritical. At the end of the extraction, the plant material, residual and exhausted of the compound of interest, is advantageously separated from the liquid phase by clarifying filtration. Even more preferably, the solvent is ethanol at 96°. We will advantageously obtain a liquid extract from the seed cake of Moringa peregrina comprising between 0.5% and 1.6% of dry matter, this dry matter being composed of at least 50% of 2,5-diformylfuran which corresponds approximately between 0.25% and 0.8% by weight of the total weight of the liquid extract.

[0035] In one embodiment of the process for obtaining according to the invention, the liquid Moringa peregrina extract obtained is purified by distillation, microfiltration, ultrafiltration and/or nanofiltration to concentrate the extract into the compound 2,5-diformylfuran of interest in relation to the organic materials also extracted, in particular to the rest of the extracted material such as fatty substances and derivatives also extracted. These purification steps make it possible to concentrate the compound of interest at the expense of other extracted compounds such as those cited as well as the solvent.

[0036] In another embodiment of the process for obtaining according to the invention, the liquid extract obtained is dried so as to obtain a dry extract of the Moringa peregrina seed cake containing more than 50% by weight of compound 2 ,5-difonnylfuran of interest relative to the total weight of the dry matter extracted.

[0037] According to an advantageous embodiment of the invention, when the solvent is ethanol, the liquid Moringaperegrina seed extract obtained is preferentially dried for example by atomization, lyophilization or zeodration so as to obtain a cake extract. of solid Moringa peregrina seeds, the ethanol being evaporated. Drying can be carried out in the presence of an organic support such as maltodextrin, cyclodextrin or inulin, or in the presence of an inorganic support such as phyllosilicate, magnesium silicate or carbonate and its salts.

[0038] The invention also relates to the extract of Moringa peregrina seeds which can be obtained by the process for obtaining it according to the invention.

The third object of the invention is a cosmetic or nutracosmetic composition comprising, as active agent, an effective quantity of an extract of Moringa peregrina seeds according to the invention, and a physiologically acceptable excipient.

The composition according to the invention can be formulated in the form of different preparations suitable for topical administration or oral administration.

According to a first variant, the different preparations are suitable for topical administration and include creams, oil-in-water and water-in-oil emulsions, milks, ointments, lotions, oils, balms, aqueous or hydro solutions. -alcoholic or glycolic, serums, powders, patches, sprays or any other product for external application such as, for example, medical devices or cosmetic-textile products.

According to a second variant, the different preparations are suitable for oral administration; the plant extract comprising the active compound 2,5-diformulfuran which can be used either in a food composition or in a food supplement. The food supplement may be in the form of capsules or soft gelatin or vegetable capsules in the context of the present invention. Said food supplement can then contain from 0.01 to 100% by weight of the plant extract. More preferably, the quantity of plant extract is 0.02 to 40% by weight, and in particular 0.2 to 20% by weight relative to the total weight of the composition.

[0042] In the context of food use, for nutritional or cosmetic purposes (cosmeto-food or nutricosmetics), the composition will advantageously be formulated in the form of a preparation suitable for oral administration. It may not include an excipient and may consist entirely of the plant extract comprising the active compound 2,5-diformylfuran.

[0043] According to a preferred embodiment, the compositions according to the invention are intended more particularly for topical administration. These compositions must therefore contain a cosmetically acceptable medium, that is to say compatible with the skin and appendages, and cover all cosmetic forms. These compositions may in particular be in the form of creams, oil-in-water emulsions, or water-in-oil or multiple emulsions, serums, solutions, suspensions, gels, milks, lotions, sticks or even powders, and suitable for application on the skin. skin, lips and/or integuments. These compositions include the excipients necessary for their formulation, such as solvents, emollients, thickeners, diluents, surfactants, antioxidants, bioactive agents, dyes, preservatives, perfumes. They can be used as a care product and/or as a skin makeup product.

[0044] The composition according to the invention may in particular consist of a composition for hair care, and in particular a shampoo, a conditioner, a treatment lotion, a styling cream or gel, a restructuring lotion for the hair, a mask , etc. The cosmetic composition according to the invention can be used in particular in treatments involving an application which is followed or not by rinsing, or even in the form of shampoo. The composition according to the invention can advantageously be used in anti-dandruff treatments. It can also be presented in the form of a dye or mascara to be applied with a brush or comb, in particular on the eyelashes, eyebrows or hair.

[0045] The compositions according to the invention further comprise any additive commonly used in the envisaged field of application as well as the adjuvants necessary for their formulation, such as solvents, thickeners, diluents, anti-oxidants, dyes, sun filters, self-tanning agents, pigments, fillers, preservatives, perfumes, odor absorbers, cosmetic or pharmaceutical active ingredients, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc.

[0046] The INCI Dictionary & Handbook (International Nomenclature of Cosmetic Ingredients (13th Ed. 2010) published by the Personal Care Products Council, Inc., Washington, D.C.) describes a wide variety, without limitation, of ingredients cosmetics and pharmaceuticals usually used in the skin care industry, which are suitable for use as additional ingredients in the compositions according to the present invention.

[0047] In all cases, those skilled in the art will ensure that these adjuvants as well as their proportions are chosen in such a way as not to harm the desired advantageous properties of the composition according to the invention.

[0048] According to an advantageous embodiment of the invention, the quantity or content of plant extract in the composition according to the invention is from 0.002 to 20% by weight, and in particular from 0.001 to 10% by weight relative to to the total weight of the composition.

[0049] Finally, the fourth object of the invention is the cosmetic or nutricosmetic use of a composition according to the invention to improve the appearance of the skin, mucous membranes and appendages, to relax, soothe and de-stress the skin and prevent and/or to fight against the skin signs of aging and/or photo-aging and prevent age spots.

[0050] According to one embodiment, the use according to the invention is more particularly intended to relax, soothe and de-stress the skin and fight against the signs of skin aging.

[0051] According to another embodiment, the use according to the invention aims to avoid the appearance of age spots.

[0052] Although the invention has been described in connection with several particular embodiments, it is obvious that it is in no way limited and that it includes all the technical equivalents of the means described as well as their combinations if these fall within the scope of the invention.

[0053] The use of the verb “include”, “understand” or “include” and its conjugated forms does not exclude the presence of other elements or other steps than those set out in a claim.

[0054] In the claims, any reference sign in parentheses cannot be interpreted as a limitation of the claim.

Examples

[0055] Example 1: Preparation of a plant extract from Morinea peresrina cake

[0056] Unshelled seeds harvested at maturity from the fruit of Moringa peregrina (Forssk.) Fiori were dried to obtain an internal humidity level of less than 8% and preferably around 6%. then pressed with a mechanical endless screw press, so as to separate the oil from the rest of the seed to obtain on the one hand the virgin oil and on the other hand a cake. The cake is then isolated in the form of sausages pre-cut into pieces of 1 to 2 cm. From the cake preheated for 10 minutes at 55°C, maceration and extraction are carried out with ethanol at 96° preheated for 10 minutes at 55°C in the ratio 25/75% (m/m), sheared in a mixer for 15 minutes then left to stir with a propeller for 2 hours at a temperature between 16°C and 30°C. We then filter with a Buchner under vacuum and we obtain a pale yellow filtrate containing 1.15% dry matter. The liquid extract obtained is subsequently named “peregrina extract according to the invention” or “peregrina extract” or “peregrina cake extract”. This liquid extract is subsequently used in the various effectiveness tests.

[0057] This peregrina extract according to the invention contains 1.15% of dry matter comprising itself (results expressed in relation to the Dry Matter (DM):

[0058] [Tableauxl]

Active compounds MS Total Compounds % Furfural 2,528 57.18 2,5-Furandicarboxaldehyde 54.66 Fats of plant origin Isopropyl myristate 1.175 42,812 Palmitic acid 4,713 Oleic acid 11,093 Trigycerides 25,831

The dry extract described above is obtained by a gravimetric method based on the mass before and after evaporation present in the liquid extract.

2,5-Fwandicarboxaldchyde or 2,5-difbrmyIfuran is the active ingredient in this extract. It was measured by chromatographic method, and more precisely by gas chromatography coupled with a flame ionization detector.

Furfural was measured using an identical method. Isopropyl myristate was measured using an identical method. Palmitic and olcic acid were measured using an identical method. Triglycerides were isolated by ultra-centrifugation.

[0059] Example 2: Effect of the peregrina extract according to the invention as an antioxidant

[0060] The objective of this study is to evaluate the modulation of the antioxidant activity by the peregrina extract in an acellular in vitro colorimetric model using the DPPH radical (2,2-diphenyM-picrylhydrazyl) as well as 'to the reference antioxidant, ascorbic acid. The method used is called inhibition. It is based on the degradation of the oxidizing radical DPPH, with a violet color absorbing at 540 nm, by a reference antioxidant, ascorbic acid. This reaction serves as a positive control and leads to the formation of the compound DPPH which is colorless or even light yellow. The peregrina extract according to the invention and the reference product “Ascorbic Acid” are placed in contact with the DPPH solution for 30 minutes at 40°C. The antioxidant activity is then evaluated by measuring absorbance at 540 nm. The modulation of this activity is expressed as a percentage of stimulation of the antioxidant activity by the active ingredient tested, with the maximum antioxidant activity obtained in the presence of ascorbic acid (T+) as a reference.

[0061] Protocol: A solution of DPPH is incubated for 30 minutes at 40°C, in the absence (control), in the presence of the deperegrina extract according to the invention (T+) and at decreasing concentrations of the sample at test. At the end of the incubation period, the antioxidant activity in the presence of the reference product and in the presence or absence of the peregrina extract was revealed by coloring after 30 minutes at 40°C. It was thus evaluated by measuring the absorbance of the reaction medium at 540 nm. For each concentration tested, the modulation of the antioxidant activity by the product tested is calculated according to the following formula.

[0062] [Math. 1]

[0063] Percentage of modulation of antioxidant activity = 100 x [(DO540 Control DO540 Product to be tested)! DO540 Reference product].

If the result is negative, the product to be tested will be considered as oxidizing; if the result is positive, the percentage will be expressed as stimulation of anti-radical activity. The results obtained are given below.

[0065] [Tables2]

DPPH inhibition Peregrina extract according to the invention 2% 36 1.0% 23 0.1% 5

[0066] Conclusion: the peregrina extract according to the invention is capable of protecting free radicals, it presents significant anti-oxidant properties from a concentration of 1%.

[0067] Example 3: Effect of the peregrina extract according to the invention as an inhibitor of metal loproteases

[0068] The objective of this study is to evaluate the modulation of the inhibitory activity of metalloproteinases by the peregrina extract according to the invention in an acellular in vitro model using a type I collagenase and a hyaluronidasc, a substrate complex and a chromophore, Ninhydrin. A buffered solution of type I collagenase and hyaluronidase reacts with a specific substrate complex and transforms it to form a compound capable of activating a chromophore by incubation at 80°C for 15 min. The activity of collagenase and hyaluronidase can thus be evaluated by measuring the absorbance at 565 nm. The sample is placed in contact with the collagenase and hyaluronidase solution at the same time as the enzyme substrate complex at 37°C for 5 minutes. The substrate transformed by the enzymes is capable of activating the chromophore by incubation at 80°C for 15 minutes. The activity of collagenase and hyaluronidase in the presence/absence of the sample is then evaluated by measuring the absorbance at 565 nm. The modulation of this activity is expressed as a percentage of inhibition or activation of the activity of collagenase and hyaluronidase in the absence of active ingredient, that is to say only in the presence of the enzyme substrate.

[0069] Protocol: A solution of type I collagenase and hyaluronidase enzymes is incubated in its substrate, for 5 minutes, in the absence or presence of the peregrina extract according to the invention tested. The solutions are then brought into contact with the chromogen, Ninhydrin, before incubation for 15 minutes at 80°C. At the end of the incubation period, the activity of the collagenase and hyaluronidase enzymes with and without test or reference product was evaluated by measuring the absorbance of the reaction media at 565 nm. For each concentration tested, the modulation of the activity of the collagenase and hyaluronidase enzymes by the product under test is calculated according to the following formula.

[0070] [Math. 2]

[0071] Percentage of modulation of the activity of the collagenase/hyaluronidase enzymes = 100 x [(DO test or reference product - DO coliagenase/hyaluronidase alone)/DO collagenase/hyaluronidase alone].

If the result is negative, the percentage is expressed as enzyme inhibition; if the result is positive, the percentage is expressed in enzyme activation. The results of the inhibition of metalloproteases by the peregrina extract according to the invention are given below.

[0073] [Tablcaux3]

Inhibition versus Control (%) Peregrina extract according to the invention 1% 87 0.5% 87 0.1% 88 0.01% 62

[0074] Conclusion: The peregrina extract according to the invention generates a strong inhibition of metalloproteinases (collagenase/hyaluronidase) of 62% at the very low levels of 0.01%. The peregrina extract according to the invention is capable of strongly inhibiting these metalloproteases and has a good potential to protect with great effectiveness the cxtra-ccllulairc matrix of the skin and, through this inhibition, it shows an anti-aging effect.

[0075] Example 4: Effect of the peregrina extract according to the invention to inhibit enzymes

Histone Deacetylases (HDACs) and Sirtnin I

[0076] The objective of this study is to demonstrate the inhibitory activity of the peregrina extract according to the invention on the enzymes HDACs and Sirtuins I. A buffered solution of HDACs & Sirtuins I reacts with a substrate for 20 minutes at 37 °C and transforms it to form a compound which colors in the presence of a developer after incubation at 37°C for 10 minutes. The maximum deacetylation activity of Sirtuins can thus be evaluated by measuring the absorbance at 405 nm. The peregrina extract according to the invention or the reference product “Trichostatin A (STA) inhibitor ΙμΜ” are placed in contact with the solution of Sirtuins at the same time as the substrate of the enzyme for 20 minutes at 37°C, the Substrate transformed by the enzyme is colored by the addition of a developer. The deacetylation activity of HDACs and Sirtuin I in the presence of the active ingredient is then evaluated by measuring the absorbance at 405 nm. The modulation of this activity is expressed as a percentage of inhibition or activation of the maximum activity of HDACs & Sirtuin I in the absence of active ingredient, that is to say only in the presence of the substrate of the HDACs & Sirtuin L enzymes.

[0077] Protocol: A solution of Sirtuin enzymes is incubated in its substrate for min in the absence (control) or presence of the reference product, or increasing concentrations of the products under test. The peregrina extract according to the invention is tested at the following concentrations: 2%; 1%; 0.1% (V/V). At the end of the incubation period, the activity of the Sirtnine enzymes with and without test or reference product was revealed by coloring using a developer solution (10 min at 37°C) and evaluated by measuring the absorbance of the reaction media at 405 nm. For each concentration tested, the modulation of the deacetylating activity of the enzymes Histone Dcsacctysases and Sirtuins I by the product under test is calculated according to the following formula.

[0078] [Math. 3]

[0079] Percentage of modulation of the activity of the Sirtuin enzymes = 100 x [(DO _4œ test or reference product) - (DO^s HDACs & Sirtuin I alone) J/DO^ Sirtuins alone.

If the result is negative, the percentage is expressed as inhibition of the enzymatic reaction; if the result is positive, the percentage is expressed as activation of the enzymatic reaction. The results of inhibition of Histone Desacetylase (HDACs) enzymes are given below.

[0081] [Tables4]

Peregrina extract according to the invention percentage Inhibition versus Control (%) 2% 20 1% ns 0.10% -18

[0082] Conclusion: At 2% the peregrina extract according to the invention shows significant inhibition of HDACs; This inhibition means the ability to promote autopiOtection of skin cells against genetic drift, particularly linked to the aging process. Thus the extract appears to be useful against one of the most common genetic deviations on the surface of the skin, namely fibrosis which manifests itself by the appearance of flesh “pimples” (fibrotic protuberance). The extract can advantageously interfere with the phenomena of fibrosis on the surface of the skin and thus prevent aging of the skin.

[0083] Example 5: Effect of the peregrina extract according to the invention to modulate the anti-inflammatory activity of the phospholypase-A2 enzyme.

[0084] The objective of this study is to evaluate the modulation of the anti-inflammatory activity of the phospholipase A2 enzyme by one or more samples in an In model.

Acellular vitro using the “SPLA2 (type V) Inhibitor Screening” analysis kit

Assay Kit”. Phospholipase A2 is a key enzyme upstream of the in21067 fiammatory process that triggers the arachidonic cascade. A buffered solution of Phospholipase A2 reacts with a specific substrate, diheptanoyl thio-PC, and transforms it to form a compound which binds to a chromogen, DTNB with stirring at room temperature. The activity of phospholipase A2 can thus be evaluated by measuring the absorbance at 413 nm. The peregrina extract according to the invention or the reference inhibitor product “Thioetheramide-PC” are brought into contact with the Phospholipase A2 solution at the same time as the enzyme substrate. The substrate transformed by the enzyme is colored using the DTNB chromogen by stirring at room temperature. The activity of the peregrina extract according to the invention or of the reference product is then evaluated by measuring the absorbance at 413 nm. The modulation of this activity is expressed as a percentage of inhibition or activation of the activity of Phospholipase A2 in the absence of active ingredient, that is to say only in the presence of the enzyme substrate (diheptanoyl thio- PC).

[0085] Protocol: A solution of Phospholipase A2 enzyme is incubated in its substrate, diheptanoyl thio-PC, in the absence or presence of the reference inhibitor and the peregrina extract according to the invention, (estimated at the following concentrations: 2%; 1%; 0.1% (V/V), then the chromogen DTNB is incorporated before incubation for 15 minutes at 25° C. At the end of the incubation period, the activity of the Phospholipase A2 enzyme with and without test or reference product is evaluated by measuring the absorbance of the reaction media at 413 nm. For each concentration tested, the modulation of the activity of the Phospholipase A2 enzyme by the test product is calculated according to the following formula.

[0086] [Math. 4]

[0087] Percentage of modulation of the activity of the Phospholipase A2 enzyme = 100 x [(OD 405 test or reference product - DO^ sPLA2 alone) /DO405 sPLA2 alone].

[0088] If the result is negative, the percentage is expressed as inhibition of the enzyme; if the result is positive, the percentage is expressed in activation of the enzyme. The results of modulation of the anti-inflammatory activity of the phospholypase-A2 enzyme are given below.

[0089] [Tables5]

Inhibition Versus Control (%) Peregrina extract according to the invention 2% 19 1% 16 0.10% 11

[0090] Conclusion: The peregrina extract according to the invention generates a slight stable inhibition of PLA2 from a dose of 0.1% but more preferably at 1 or 2%. This means that the peregrina extract according to the invention has the capacity to reduce the arachidonic cascade very upstream! inflammation cascade; This extract therefore has good soothing or relaxing potential on the skin.

[0091] Example 6: Effect of the peregrina extract according to the invention to inhibit the action of endothelin 1

[0092] Endothelin is the most powerful vasoconstrictor known in the human body. On the other hand, the decrease in endothelin is also known to create a vasodilator effect [Hirata Y. et al., 1988, Cellular mechanism of action by a novel vasoconstrictor endothelin in cultured rat vascular smooth muscle cells, Biochemical and Biophysical Research Communication 154:3, p. 868-875,] [Shalinkümar P. et al., 2018, H2S Mediates the Vasodilator Effect of Endothelin-1 in the Cerebral Circulation. American Journal of Physiology. Heart and Circulatory Physiology 315, p. 1759-1764],

[0093] The objective is to measure type 1 endothelin in human microvascular endothelial cells after exposure for 24 hours to the peregrina extract according to the invention.

[0094] Protocol: The human microvascular endothelial cells were supplied by the company PELOBiotech and cultured in 96-well plates according to the supplier's production procedures. This involves letting the extracts act at different concentrations on endothelial cells at 80% confluence for 24 hours, then quantifying endothelin 1 using the PicoKine ELISA kit (EDN1) in the cell supernatants. A viability test is carried out beforehand to define the non-toxic doses to be used when measuring endothelin 1. The negative control is carried out using the cells in culture medium without treatment. The positive control in the viability test is 0.5% SDS. All conditions are prepared in culture media, and the cells are then incubated at 36.5°C/5% CO2 for 24 hours.

a) Application of the solutions to be tested on endothelial cells:

The products to be tested are placed in contact with the subconfluent endothelial cells in 96-well plates. For each concentration the test is carried out on 3 wells. The plates are incubated for 24 hours ± 1 hour at 36.5°C 15% CO2.

b) Viability test;

Cell viability is evaluated by the MTT method on the cells after incubation with the products. After 24 hours of incubation, the supernatants are collected and stored at -20°C for assays. The wells are then rinsed I times with 200 μ-L of PBS. In each well, 50 μL of MTT solution at 0.5 mg/ml are added: incubation for 3 hours at 36.5°C/5% CO2. In each well 100 μL of isopropanol are added. After homogenization, an absorbance reading is taken at 550 nm. For each condition, the ratio of the average optical densities of the cells to the average optical densities of the negative controls will determine the viability rate.

c) Dosage of endothelin 1:

The assay is carried out using the ELISA kit.

[0095] [Tableauxô]

Extract concentration Cell growth versus control (%) Endothelin 1 versus control (%) Endothelin 1 versus control (pg/ml) Peregrina extract according to the invention 5% -22.31 -53.21 -71.8 1% 0 -11.61 -15.66 0.10% -5.77 -25.11 -33.88

[0096] Conclusion: The viability test carried out at the end of treatment did not show any toxic effect for the concentrations tested. Endothelin 1 is assayed in cell supernatants at non-toxic concentrations. The quantity of endothelin 1 for each condition is measured using the ELISA kit. For the negative control cells, the values are around 134.94 pg/ml. For cells treated with different concentrations of extracts, the values are from 63.14 pg/ml (with 5% of the extract according to the invention) to 101.06 pg/ml (with 0.1% of the extract according to the invention) , which shows very significant inhibitions from 0.1% of the extract according to the invention with approximately 25% inhibition of type 1 endothelin production and up to 53% inhibition with 5% of the extract according to the invention. 'invention.

[0097] Example 7: Effect of the peregrina extract according to the invention to stimulate the activity of telomerase

[0098] Telomeres are DNA protective complexes located at the ends of linear chromosomes which promote chromosomal stability. Telomere shortness in humans is emerging as a prognostic marker for disease risk and progression, and premature mortality in many types of cancers, including breast, prostate, colon, bladder. , head and neck, lung and kidney cells [Ornish D., 2008, Increased Telomerase Activity and Comprehensive Lifestyle Changes: a Pilot Study, Lancet Oncology 9, p. 1048-1057], Telomere shortening is counteracted by the cellular enzyme telomerase.

[0099] The objective of the study is to evaluate the effect of the compound named “Peregrina extract according to the invention” on telomerase activity in a model composed of adult human keratinocytes at a low level of passage in monolayer culture.

[0100] Protocol: Human keratinocytes were obtained from a 49-year-old donor. To perform the experiments, keratinocytes were used at a low passage level (i.e., passage #2 of cell isolation). Cells were grown in monolayer until reaching approximately 75% confluence before being used in the experiment.

Reference product: FK228 at 100 ng/ml was used as a reference inducer of telomerasel activity.

Incubation protocol: The cells were incubated for 24 hours in the absence (control) or in the presence of reference product or increasing concentrations of test compounds such as the peregrina extract according to the invention at: 0.5; 1 and 5% (v/v).

The peregrina extract according to the invention is diluted directly in the incubation medium to reach the different concentrations described above.

Assessment of effects:

- Protein measurement

At the end of the incubation period, total cell proteins were extracted from the cells and measured using a spectro-colorimetric method (Bradford method). This measurement makes it possible to determine the exact volume of extract to use during the telomerase activity measurement, in order to maintain the same quantity of proteins (containing telomerase) for all conditions tested at the PCR step.

- Measurement of telomerase activity

At the end of the incubation period, telomerase was extracted from the cells and its activity was determined using a specific and sensitive kit. The principle of the telomerase kit is to measure telomerase activity coupling a PCR step (in which telomerase works on its elongation activity) with an ELIS A step allowing the semi-quantitative determination of quantities of elongation product. telomerase.

- Statistics

Results are expressed in arbitrary units for telomerase activity level (mean ± S.D). The level of significance between “vehicle” and “reference product” was assessed using a Student’s t test (*: p < 0.05). The level of significance between “control” and “test compound” was assessed independently for each product by a one-way analysis of variance (one-way ANOVA) followed by a Holm-Sîdak test (*: p < 0.05).

The peregrina extract according to the invention, tested at 0.5% and 1% (v/v), did not significantly modulate telomerase activity compared to “Control”. Tested at 5% (v/v), the peregrina extract according to the invention significantly increased telomerase activity by 18.9% (pcO.OOl), in comparison with “Control”. The reference product, named “FK228”, tested at 100 ng/ml, significantly increased telomerase activity by 28.0% (p <0.01). This result was expected and validates the experiment. The results of stimulation of telomerase activity are given below,

[0101] [Tables?]

Extract concentration Cell growth versus control (%) Telomerase activity versus control (%) Peregrina extract according to the invention 5.00% +5.8 +18.90 1.00% That. -2.2 +4.00 0.50% -2.6 +2.30

[0102] Conclusion the peregrina extract according to the invention, tested at 0.5% and 1% (v/v), did not significantly modulate telomerase activity compared to “Control”. Tested at 5% (v/v), the peregrina extract according to the invention significantly increased telomerase activity by 18.9% (p<0.001) in comparison with “Control”. The extract according to the invention acts directly on this enzymatic pathway which builds protective telomere at the ends of chromosomes and slows down the natural aging of genetic material. The extract according to the invention therefore makes it possible to produce an anti-aging effect on the chromosomes.

[0103] Examples 2 to 7 make it possible to demonstrate that the peregrina extract according to the invention has anti-aging, anti-stress and relaxing properties which demonstrates the profile of a good skin protector.

[0104] Example 8: Effect of the perggrma extract according to the invention to stimulate the activity of liUymsmase

[0105] The objective of this study is to evaluate the activity on the Tyrosinase enzyme of the peregrina extract according to the invention in an acellular in vitro model using a tyrosinase enzyme of fungal origin (Sigma Aldrich ref. . T3824), to its substrate LTyrosine (Sigma Aldrich ref. T3754) and a reference inhibitor hydroquinone (Sigma Aldrich ref. H17902, Inhibitor = Hydroquinone 2.5 mM). A buffered Tyrosinase solution reacts with a 2.5 mM L Tyrosine substrate for 60 minutes at 23°C and transforms it to form a colored compound. The maximum activity of Tyrosinase can thus be evaluated by measuring the absorbance at 475 nm. The peregrina extract according to the invention or the reference product “Hydroquinone” are placed in contact with the Tyrosinase solution at the same time as the substrate of the enzyme for 60 minutes at 23°C, the substrate transformed by the enzyme is naturally colored. Tyrosinase activity in the presence of the active ingredient is then evaluated by measuring the absorbance at 475 nm. The modulation of this activity is expressed as a percentage of inhibition or activation of the maximum activity of Tyrosinase in the absence of active ingredient, that is to say only in the presence of the enzyme substrate (L Tyrosine). .

[0106] Protocol: A solution of Tyrosinase enzyme is incubated in its substrate L Tyrosine for 60 minutes in the absence (control) or presence of the reference product, or increasing concentrations of the peregrina extract according to the invention / concentrations; 2%; 1%; 0.1% (V/V). At the end of the incubation period, the activity of the Tyrosinase enzyme with and without test or reference product was evaluated by measuring the absorbance of the reaction media at 475 nm. For each concentration tested, the modulation of the activity of the Tyrosinase enzyme by the product under test is calculated according to the following formula:

[0107] [Math. 5]

[0108] Percentage of modulation of the activity of the Tyrosinase enzyme = 100 x [(OD475 test or reference product) - (DO475 Tyrosinase alone)] /DO475 Tyrosinase alone.

[0109] If the result is negative, the percentage is expressed as inhibition of the enzyme; if the result is positive, the percentage is expressed in activation of the enzyme. The stimulation results on tyrosinase activity are given below.

[0110] [Tables8]

Activation Versus Control (%) Peregrina extract according to the invention 2% 65 1% 37 0.10% 8

[0111] Conclusion: The peregrina extract according to the invention is capable of lowering the basal activity level of tyrosinase, which makes it possible to indicate that this extract has the capacity to increase one of the natural protections of the skin , protection against ultraviolet rays.

[0112] Example 9: Effect of the peregrina extract according to the invention to inhibit the production of melanin

[0113] The objective of this study on human cell cultures is to bring together all the data used, as well as the results obtained, for carrying out the melanin modulation test on human melanocytes after exposure for 5 days to 'peregrina extract according to the invention.

Protocol: Human melanocytes are cultured in 96- and 24-well plates.

this involves letting the peregrina extract according to the invention act at concentrations of 5%; 2%; 1% and 0.1% on confluent melanocytes for 5 days. A pre-test of viability with MTT after 24 hours makes it possible to evaluate cytotoxicity and to choose the concentrations for the melanin modulation test. This modulation is evaluated by measuring melanin in cell lysates after 5 days of exposure to the extracts. The negative control is carried out using the cells in culture medium without treatment. The positive control for the viability test is 0.5% SDS. For testing melanin modulation, media with and without α-MSH are used as negative controls.

All conditions are prepared in culture media, and the cells are then incubated at 36.5°C/5% CO2 for 24 hours for the cytotoxicity test and 5 days for the melanin assay.

a) Application of the solutions to be tested on the melanocytes: The concentrations to be tested are brought into contact with the confluent melanocytes in the 96 (cytotoxicity test) and 24 (melanin dosage) well plates. For each concentration the test is carried out on 3 wells. The plates are incubated for 24 + 1 hours and 5 days at 36.5°C / 5% CO2.

b) Viability test: Cell viability is evaluated by the MTT method on the cells after 24 hours of incubation with the products. After 24 hours of incubation, the planned wells are rinsed once with 200 μΕ of PBS. In each well, 50 μΐ of MTT solution at 0.5 mg/ml are added and the incubation is carried out for 3 hours at 36.5°C/5% CO2. In each well 150 μΐ of isopropanol are added. After homogenization, an absorbance reading is taken at 550 nm. For each condition, the ratio of the average optical densities of the cells to the average optical densities of the negative controls will determine the viability rate.

A viability threshold value of 70% relative to the negative control value is used to classify the substances tested as cytotoxic or non-cytotoxic. A “non-cytotoxic” classification is given on the in vitro results for a viability > 70% and a “cytotoxic” classification for a viability < 70%.

The 5% concentration of the extract according to the invention was found to be cytotoxic at 5% under the test conditions. The concentrations 2%, 1% and 0.1% are therefore used for the melanin modulation test. The quantity of melanin present in the cells is measured after cell lysis. The results on the inhibition of melanin production are given below.

[0114] [Tables9]

Extract concentration Cell growth versus control (%) Inhibition Melanin versus control (%) Melanin content (pg/ml) Peregrina extract according to the invention 2% -3.41 +16.50 152 1.0% +13.46 +65.90 62 0.10% + 13.29 +71.40.. 52 -

Conclusion: the peregrina extract according to the invention inhibits the production of melanin in

[0115]

[0116] cellulo which gives it a protective property for the skin. This inhibition also shows a relaxing effect on melanocytes because the basal level is lower than the control (absence of extract according to the invention in the culture medium); It is recalled that the production of melanin is a response to cellular stress. Thus, peregrina according to the invention demonstrates the ability to prevent aging spots.

Example.10: Analytical characterization of the peregrina extract according to the invention versus the oto'fera extract with an identical extraction process

On the basis of Moringa peregrina cake and that of Moringa oleifera, the extraction process according to the invention described in Example 1 was applied. The comparative composition of the extracted ingredients on the basis of dry matter is given below.

[Tables 10]

Compounds Oleifera (%) Peregrina (%) Acetic acid 1,431 - 1 -hydroxy-2~propanone 1,962 - Furfural 6.140 2,528 2,5-Furandicarboxaldehyde (DFF) 0.823 55,660 Isopropyl myristate 0.128 1.175 Methyl palmitate 0.230 — Palmitic acid 10,356 4,713 Ethyl palmitate 0.399 — Methyl linoleate 0.428 — Methyl oleate 2,443 — Oleic acid 47,844 11,093 Ethyl oleate 2,675 — Stearic acid 5.064 — Bis 2-ethyl hexyl hexanedioate 1,688 — Methyl 7-oxo-dehydroabietate 0.327 — 2-ethyl-hexyl 1,4-terephthalate 0.655 — Total 84,317 74.169

It appears that the two extracts have a very different molecular profile. Moringa oleifera extract contains less than 1% DFF while Moringa peregrina extract contains more than 50%.

[0117] Example 11: Comparative test with Moringa oleifera for in tuba collagenase test [0118] The objective of this study is to evaluate the modulation of the inhibitory activity of Metalloproteases by the peregrina extract according to the invention in a model acellular in vitro using a type I collagenase and a hyaluronidase. a substrate complex and a chromophore, Ninhydrin. A buffered solution of Collagenase type I and hyaluronidase reacts with a specific substrate complex and transforms it to form a compound capable of activating a chromophore by incubation at 80°C for 15 min. The activities of collagenase and hyaluronidase can thus be evaluated by measuring the absorbance at 565 nm. The sample is placed in contact with the Collagenase and Hyaluronidase solution at the same time as the enzyme substrate complex at 37°C for 5 minutes. The substrate transformed by the enzymes is capable of activating the chromophore by incubation at 80°C for 15 minutes. The activity of Collagenase and Hyaluronidase in the presence/absence of the sample is then evaluated by measuring the absorbance at 565 nm. The modulation of this activity is expressed as a percentage of inhibition or activation of the activity of Collagenase and Hyaluronidase in the absence of active ingredient, that is to say only in the presence of the enzyme substrate.

[0119] Protocol: A solution of Collagenase type I and Hyaluronidase enzymes is incubated in its substrate, for 5 minutes, in the absence or presence of the peregrina extract according to the invention tested. The solutions are then brought into contact with the chromogen, Ninhydrin, before incubation for 15 minutes at 80°C. At the end of the incubation period, the activity of the Collagenase and Hyaluronidase enzymes with and without test or reference product was evaluated by measuring the absorbance of the reaction media at 565 nm. For each concentration tested, the modulation of the activity of the Collagenase and Hyaluronidase enzymes by the product under test is calculated according to the following formula.

[0120] [Math. 6]

[0121] Percentage of modulation of the activity of the Collagenase/Hyaluronidase enzymes = 100 x [(DO test or reference product - DO Collagenase/Hyaluronidase alone)/DO Collagenase/Hyaluronidase alone].

[0122] If the result is negative, the percentage is expressed as enzyme inhibition; if the result is positive, the percentage is expressed in enzyme activation. The result of the inhibition of Metalloproteases is given below.

[0123] [Tableauxll]

Inhibition versus Control (%) Peregrina extract according to the invention 1% 87 0.5% 87 0.1% 88 0.01% 62

[0124] Conclusion: The peregrina extract according to the invention generates a strong inhibition of

Metalloproteases (Collagenase / Hyaluronidase). It is capable of inhibiting 88% of these

Metalloproteases from a concentration of 0.1% and has a good potential to protect with great efficiency the extra-cellular matrix of the skin and, through this inhibition. it shows an anti-aging effect.

[0125] To be compared with the extract according to patent FR2946879 from Pierre Fabre, of which here are the results according to the same test.

[0126] [Tables 12]

Inhibition versus Control (%) Moringa oleifera extract according to Pierre Fabre patent 1% Concentration not compatible with testing system 0.5% 4 0.1% 24 0.01% 42

[0127] Conclusion the extract according to the Pierre Fabre patent shows a slight inhibitory action on collagenase activity in inverse dose dependence with an inhibition peak of 42% for all concentrations combined compared to an inhibition peak of 100% for the peregrina extract according to the invention.

[0128] The anti-aging activity on this parameter appears different and new compared to the effects observed with the extract according to the Pierre Fabre patent.

[0129] Example 12: Comparative tests for anti-stress activity fby inhibition of PLA2'). [0130] For anti-stress activity by inhibition of PLA2 on the skin which provides a soothing/anti-stress orientation with an anti-aging effect. two additional PLA2 in tubo tests were conducted; one with the process according to the invention carried out on oleifera cake (extract made identically to that on peregrina in the process according to the invention), one with the extract corresponding to document FR2946879 by Pierre Fabre, another still corresponding to patent FR3076460 from BASF Beauty Care Solutions with the Purisoft® product and finally a last one with document FR2825267 from Chuun & Thurol.

[0131] The objective of this study is to evaluate the modulation of the anti-inflammatory activity of the phospholipase A2 enzyme by one or more samples in an acellular In Vitro model using the “SPLA2” analysis kit (type V) Inhibitor Screening Assay Kit”.

[0132] A buffered solution of Phospholipase A2 reacts with a specific substrate, diheptanoyl thio-PC, and transforms it to form a compound which binds to a chromogen, DTNB, with stirring at room temperature. The activity of phospholipase A2 can thus be evaluated by measuring the absorbance at 413 nm.

[0133] The products “Extraitperegrina according to the invention” or the reference product in21067 inhibitor “Thioetheramide-PC” are placed in contact with the solution of Phospholipase A2 at the same time as the substrate of the enzyme. The substrate transformed by the enzyme is colored using the chromogen DTNB by stirring at room temperature.

The activity of Phospholipase A2 in the presence/absence of the product “Peregrina extract according to the invention” or of the reference product is then evaluated by measuring the absorbance at 413 nm.

[01.34] The modulation of this activity is expressed as a percentage of inhibition or activation of the activity of Phospholipase A2 in the absence of active ingredient, that is to say only in the presence of the enzyme substrate ( diheptanoyl thio-PC).

[0135] The inhibitor “Thioetheramide-PC” at the concentration of Img in 100 μ1 is the reference product (active control) in this study, the latter inhibits the activity of PLA2 by 93%, which validates the test.

[0136] A Phospholipase A2 enzyme solution is incubated in its substrate, diheptanoyl thio-PC, in the absence or presence of the reference inhibitor and the product “Peregrina extract according to the invention” tested then the chromogen DTNB is incorporated before incubation for 15 minutes at 25°C.

[0137] At the end of the incubation period, the activity of the Phospholipase A2 enzyme with and without test or reference product was evaluated by measuring the absorbance of the reaction media at 413 nm. For each concentration tested, the modulation of the activity of the Phospholipase A2 enzyme by the test product is calculated according to the following formula:

[0138] [Math. 7]

[0139] Percentage of modulation of the activity of the Phospholipase A2 enzyme = 100 x [(OD405 produced in the test or reference - DO405 sPLA2 alone) /DO405 sPLA2 alone]

[0140] If the result is negative, the percentage is expressed as inhibition of the enzyme; if the result is positive, the percentage is expressed in activation of the enzyme.

[0141] [Tables! 3]

Peregrina extract according to 1 invention 1% 19 0.1% 16 0.01% 11

The consistency and substantially dose-dependent activity of this peregrina extract according to the invention on the inhibition of PLA2 highlight a soothing activity by reducing the intensity upstream of the arachidonic cascade. Such a soothing action very upstream of the arachidonic cascade reduces the impact of basal physiological stress. The peregrina extract according to the invention is therefore an anti-stress.

[0142] With the protocol extract from patent FR2946879 by Pierre Fabre, the following results are obtained.

[0143] [Tables 14]

Excerpt from Pierre Fabre 1% 0 0.1% 16 0.01% 8

At 1%, which corresponds to its minimum dosage, the extract does not show any activity. When diluted, an activity appears but the lack of dose-dependent effect does not make it possible to validate a specific and reliable action of the extract on the inhibition of this enzyme.

[0144] With the protocol extract from patent FR3076460 of BASF Beauty Care Solutions with the product Purisoft® LS9726, the following results are obtained.

[0145] [Tables 15]

BASF extract Purisoft LS9726 1% 10 0.1% 12 0.01% 6

This extract shows a slight inhibitory action which does not reach the maximum observed at 1% with the peregrina extract according to the invention, namely 19% inhibition for the extract according to the invention against 10% inhibition at 1% for this extract.

[0146] With the protocol extract from patent FR2825267 of Chuun & Thurot we observe the following results.

[0147] [Tables 16]

Extract from Chuun & Thurot 1% 0 0.1% 0 0.01% 0

This extract does not show any inhibitory action on this enzyme.

[0148] With the protocol extract resulting from the process according to the present invention but applied to Moringa oieifera cake instead of Moringa peregrina cake, we observe the following results.

[0149] [Tables 17]

Moringa oleifera extract 1% Undosable 0.1% 0 0.01% 6

This extract does not show any significant or stable inhibitory action on this enzyme. Conclusion: only the peregrina extract according to the invention demonstrates significant inhibitory activity on the PLA2 enzyme.

[0150] Example 13; Formulation of a makeup product

[0151] [Tablesl8]

Ingredients % Water QSP Caprylic / Capric triglyceride 19.0000 Gum Acacia Senegal 7.0000 Charcoal 6.0000 Glycerin 5.0000 Propanediol 5.0000 Bentonite 3.1500 Cetearyl glucoside 3.0000 Cetearyl alcohol 3.0000 Benzyl alcohol 1.0000 Peregrina extract according to invention 2.0000 Cellulose gum 0.8000 Xanthan gum 0.1750 Citric acid 0.1750

[0152] Example 14: Formulation of a cleansing product

[0153] [Tablesl9]

Ingredients % Water QSP Sodium coco sulfate 5.0000 Sodium cocoyl isethionate 4.0000 Bentonite 3.7800 Caprylic i Capric triglyceride 2.0000 Peregrina extract according to the invention 1.0000 Gluconolactone 0.7500 Sodium benzoate 0.5450 Scent 0.5000 Xanthanc gum 0.2700 Sodium stearoyl glutamate 0.2250 Citric acid 0.2250 Calcium gluconate 0.0050

[0154] Example 15: formulation of a care product [anti-aging anti-stress cream)

[0155] [Tables20]

Ingredient % Water QSP Caprylic / Capric triglyceride 18.0000 Bentonite 4.2000 Cetearyl alcoho] 1.5000 Peregrina extract according to invention 5.0000 Gluconolactone 0.7500 Sodium benzoate 0.5450 Xanthan gum 0.5000 Scent 0.5000 Sodium stcaroyl glutamate 0.2500 Citric acid 0.2500 Calcium gluconate 0.0050

[0156] Example 16: oral formulation for nutricosmetics

[0157] A 1 g tablet for soothing/anti-stress activity comprises: 3% dry extract according to the invention (containing 0.6% of 2,5-diformylfuran on an inulin support) + 47% of calcium carbonate containing 200 IU of vitamin D + 25% magnesium gluconate + 22% inulin + 3% magnesium stearate.

[0158] Example 17: Toxicological tests of the peregrina extract according to the invention Preparation of the peregrina extract in accordance with example 1;

Seeds of Moringa peregrina (Forssk.) Fiori, collected when the fruit was ripe, were dried to obtain an internal humidity level of around 6%, then pressed with a mechanical worm press, so as to separate the oil from the rest of the seed to obtain on the one hand virgin oil and on the other hand a cake. The cake is then isolated in the form of sausages pre-cut into pieces of 1 to 2 cm. From the cake we carry out maceration and extraction with ethanol at 96° preheated for 10 minutes at 55°C in the ratio 25/75% (m/m), We shear in a mixer for 15 minutes then we leave to shake at the propeller for 2 hours at 20°C. We then filter with a Buchner under vacuum and we obtain a pale yellow filtrate containing 1.15% dry matter. This filtrate is used pure in the tests which follow.

[0159] Determination of mutagenic activity on the bacterial strain Salmonella ty21067 ohimurium (TA 100) - Bacterial reverse mutation test

The test took place in 3 main phases:

- A preliminary experiment is carried out in order to evaluate the cytotoxicity of the element to be tested and to select the dose range for subsequent experiments, - A first genotoxicity experiment (Test 1), with and without metabolic activation, with direct incorporation of the test system and the test (or controls) on minimal agar, over the dose range defined by the preliminary study,

- A second experiment (Test 2), with pre-incubation of the test system and test item (or controls), with and without metabolic activation, with dose levels defined by the study manager after analysis of the results of the first experiment. This second experiment was carried out in order to confirm or complete the results of the first, in particular when equivocal or negative results were obtained.

Dilutions of test items were prepared in water for analysis.

[0160] The cytotoxicity test was carried out on the S. typhimurium TA 100 strain at concentrations of 5000, 1600, 500, 160 and 50 pg/plate, with and without S9-Mix.

The reagents used for the preparation of S9-Mix were prepared according to the following instructions:

[0161] [Tables21]

Final concentration MgC12 (0.4 M) + KC1 (1.65 M) 8mM + 33mM Glucose 6 Phosphate (0.2 M) 5mM N ADP (0.1 M) 4mM Phosphate buffer for S9-Mix (pH 7.4 -0.2 M) 0.1M S9 fraction 10% Water Adjust for final concentration

[0162] The bacteria were exposed to the test extract with and without a metabolic activation system. The metabolic system used is a cofactor-enhanced post-mitochondrial fraction (S9). This S9 fraction, a microsomal fraction of Sprague Dawley rat liver homogenate treated with an enzyme inducer, is prepared according to Maron, D.M. and Ames, B.N. (1983) and was supplied by MOLTOX TM. It is stored at a temperature below -70 °C. The S9 microsomal fraction was used at a concentration of 10% in S9-Mix. The protocol applied was as follows:

• In 3 hemolysis tubes, was introduced:

o dosage without metabolic activation:

• 0.1 ml of the different concentrations of the test elements, • 0.5 ml of sterile 0.2 M phosphate buffer, pH 7.4, • 2 ml of top agar for S. typhimurium, * 0.1 ml of bacterial inoculum (TA100), o dosage with metabolic activation:

• 0.1 ml of the different concentrations of the test elements, • 2 ml of top agar for 5. typhimurium, • 0.1 ml of bacterial inoculum (TA100), • 0.5 ml of S9-Mix.

• Mix and pour onto the surface of the background agar previously distributed in Petri dishes, • Incubate at 37°C±2°C for 48 to 72 hours.

These assays were carried out for each test: preliminary cytotoxicity test, test 1 and test 2. The untreated control, the negative controls and the positive controls carried out during the pre-incubation method were incubated for 20 to 30 minutes at 37°C ± 2°C before pouring the top agar. The protocol applied was as follows:

• In 4 fractions of 2 ml of top agar for S, typhimurium, introduce: o 0.1 ml of 0.2 M phosphate buffer, pH 7.4, o 0.1 ml of solvent, o 0.1 ml of S9-Mix.

o 0.1 ml of the preparation of the element to be tested at the highest concentration, * A fraction of 2 ml of top agar for A typhimurium is used to check its sterility.

• Mix and pour onto the surface of the background agar previously distributed in Petri dishes.

• Incubate at 37°C + 2°C for 48 to 72 hours.

• The test is carried out in triplicate.

• No bacterial growth should be observed.

For at least 5 concentrations of the extract to be tested, a test without metabolic activation and a test with metabolic activation were carried out.

[0163] Expression of the results and interpretation

Many criteria make it possible to determine whether a result is positive, in particular an increase in the number of revenants correlated with the dose of item to be tested, or a reproducible increase in the number of revenants at one or more concentrations, with or without metabolic activation.

- The element to be tested is considered mutagenic if, at the end of the verification steps, a dose-effect relationship has been obtained, in a reproducible manner, on one or some of the 5 strains with and/or without metabolic activation. Mutagenicity is only taken into account for a given concentration when the number of revertants is at least twice the spontaneous reversion rate for strains TA98, TA 100 and TA 102 (R > 2) and three times the spontaneous rate. reversion for strains TA 15 35 and TA 1537 (R > 3).

- The test item is considered non-mutagenic if, after test 1 and test 2, the revertant rate has always remained less than twice the spontaneous reversion rate for all concentrations of the test item tested, 10 for strains TA98, TA 100 and TA102 (R <2) and less than three times the spontaneous reversion rate for strains TA 1535 and TA 1537 (R <3), with and without metabolic activation, and provided that It has been verified that the absence of the mutagenic effect is not linked to the toxicity of the concentrations tested.

The preliminary study showed no cytotoxicity of the test element; consequently, this concentration range was used for genotoxicity test 1. [0164] Depending on the result obtained for ballast 1, it was decided to use the same dilution range for test 2. analysis of the revertants shows that: - No cytotoxic effect was observed, - No concentration of the extract to be tested showed an R ratio greater than 20 or equal to at least twice the spontaneous reversion rate for TA98,

TA 100 and TA 102 and three times the spontaneous reversion rate for TA 1535 and TA1537, with and without metabolic activation,

No dose response was observed regardless of test system or test conditions.

[0165] In view of the results obtained during this study, the peregrina extract according to Example 1 can be considered as having no mutagenic or promotagenic activity.

[0166] 3T3 NRU in vitro phototoxicity test

The principle of the test is based on the comparison of the cytotoxicity of the peregrina extract according to Example 1 in the presence and absence of a non-cytotoxic dose of UVA, on cells in culture. Cytotoxicity is evaluated by determining cell viability using a vital dye: neutral red, 24 hours after treatment with the reference elements and the extract of M. peregrina with or without UV irradiation. The cells used are Balb/c 3T3 lineage mouse embryo fibroblasts clone 31 (ATCC — CCL163). The positive control is a solution of chlorpromazine (CAS number: 69-09-0). The negative control is a diluent of the tested and reference extract (buffered saline solution +/- 1% solvent). The peregrina extract was tested at 8 concentrations on at least four culture wells per concentration studied, in the presence or absence of UVA. The fibroblasts were trypsinized and two 96-well culture plates were seeded at 100 μ! of a cell suspension at 2.10 ³ cells/ml (i.e. 2.10 ^cells per well) in complete culture medium.

The seeded plates were incubated in the oven for 24 hours at 37°C, 5% CO ₂ . At the end of the incubation, the semi-confluence of the cell layer was checked. The dilutions were prepared just before their deposition on the cells. The pH of the highest concentration was measured; it is between 6.5 and 8. The culture medium was eliminated, each well was carefully rinsed beforehand with 150 μΐ of PBS maintained at room temperature before being treated with 100 μΐ of each dilution of the extract or reference . The culture plates were incubated in the dark for 1 hour plus or minus 5 minutes at 37°C and 5% CO ₂ . Irradiation was carried out using a BIO SUN solar irradiator (Vilber Lourmat RMX3W). The BIO SUN is a system that controls UV irradiation using a programmable microprocessor. The system continuously tracks the UV light emission. Irradiation stops automatically when the energy delivered is equal to the programmed energy. The spectral irradiation of the device under test was measured in the wavelength range 250 to 700 nm with a calibrated spectroradiometer. One of the 2 plates was irradiated with its lid at room temperature, and the other plate was protected from UVA and kept at room temperature during the irradiation. After irradiation, the treatment medium was aspirated and the cells were rinsed. Then without μΐ of complete culture medium were added gently and the plates were incubated for 18 to 22 hours at 37°C and 5% CO ₂ . The next day, cell viability (growth, morphology, integrity of the monolayer) was evaluated by observations on a phase contrast microscope. The culture medium was removed, each well was previously rinsed and maintained at room temperature before being treated with 100 μΐ of the staining solution. The plates were returned to the incubator for 3 h under the same conditions. The dye solution was removed and the cells were rinsed, then the rinsing solution was removed and 150 μΐ of desorption solution was added to each well. The plates were shaken until the crystals were completely solubilized. Absorbances were measured at 450 nm.

[0167] Validation of the test:

The sensitivity of the cells to UVA is monitored approximately every 10 passages, by evaluating their viability after exposure to increasing doses of irradiation. The cells are cultured at the density used in the assay. They are irradiated the following day at doses of 2.5 and 9 J/cm ² and cell viability is determined one day later using the NRU test. The cells meet the quality criteria if their viability after irradiation at 5 J/cm2 UVA is greater than or equal to 80% of the viability of the controls kept in the dark: at the highest dose of 9 J/cm2 UVA, the Viability must be at least equal to 50% of that of controls kept in the dark. [0168] Results:

The negative control has an absorbance greater than or equal to 0.4. Chlorpromazine, 5 positive control, has an IC ₅₀ between 0.1 and 2 μg/ml in the presence of UVA and between 7 and 90 pg/ml in the absence of UVA. These results allow the test to be validated. The concentration of peregrina cake extract giving 50% cell death in the presence or absence of UVA cannot be estimated. Mortality never reached 50%. The concentration of peregrina cake extract giving 50% cell viability in the presence or absence of UVA cannot be estimated. Viability is always above 50%.

Conclusion: Under the experimental conditions retained, peregrina cake extract can be considered non-phototoxic.

[0169] Evaluation of the ocular irritant potential by the study of in vitro cytotoxicity according to the neutral red release method on STRC cell line

This in vitro study is based on the evaluation of the cytotoxicity of peregrina cake extract by determining the concentration leading to 50% cell mortality (IC ₅₀ ) on a cell monolayer using the release technique. neutral red. The cells used are SIRC rabbit corneal fibroblasts (ATCC - CCL60) free of mycoplasmas.

The peregrina extract was diluted in 25% and 50% physiological saline. The fibroblasts were trypsinized and two 24-well culture plates were seeded with 1 ml of a cell suspension at 2.10 ⁵ cells/ml in complete culture medium. The seeded plates were incubated in the oven overnight at 37°C, 5% CO ₂ . 25 At the end of the incubation, the confluence of the cell layer was checked. The staining solution was prepared at 0.5 mg/ml in complete culture medium. The culture medium was removed, 1 ml of the dye solution was placed in each well. The plates were returned to the incubator at 37°C, 5% CO2 for 3 hours +/- 15 minutes. After this contact time, the dye solution was removed and replaced with 1 ml of complete culture medium per well. The plates were kept at room temperature for at least 30 minutes to stabilize the system before contact with the extract or reference. Each well was rinsed with 2 ml of PBS, maintained at room temperature, then 500 μΐ of each dilution of peregrina or reference extract were deposited in contact with the cell layer. The contact time was 35 to 60 seconds (30 seconds for the positive control). The treatment was carried out well by well with the stopwatch triggered at the time of deposition of the peregrina extract or the reference. The plate was shaken manually throughout the treatment. After 55 seconds (or 25 seconds for the positive control, the dilution was aspirated. At exactly 60 or 30 seconds, 5 successive rinses were carried out (5x2 ml of PBS maintained at room temperature). The supernatant was aspirated after each rinse. and after the last rinse the wells remained devoid of medium while awaiting the revelation phase. After complete treatment of the culture plate, 1 ml of the desorption solution was deposited in each well. The plate was shaken for approximately 15 minutes until homogeneous coloring was obtained. The solutions obtained for each culture well were taken and distributed into 2 wells of a 96-well plate, i.e. 150 μΐ/well.

[0170] Results;

The concentration of the peregrina extract giving 50% cell death was evaluated at >50%. The percentage of cell death at 50% of peregrina extract was estimated at 17%

Conclusion: Under the experimental conditions chosen, the cytotoxicity of peregrina cake extract can be classified as negligible cytotoxicity.

15[0171] Evaluation of the skin compatibility of a peregrina extract after single application under an occlusive dressing for 48 hours under dermatological control.

The aim of this study is to evaluate the degree of skin compatibility of the peregrina extract by patch test, carried out on the anterolateral side of the arm for 48 hours; and generally evaluate the capacity of the peregrina extract to maintain the skin in good condition. 10 holy volunteers, female or male aged 18 to 65 years, having neither dry nor sensitive skin and free from any dermatological lesion on the treatment area were to be included in the study. The skin compatibility of the peregrina extract, prepared in the form of a lotion with 5% of the peregrina extract according to Example 1 and 95% of a Propanediol/Sorbitol mixture, was evaluated.

48 hours after initial application between 30 to 40 minutes after removal of the dressing. Skin reactions (erythema and edema) were counted from 0 to 3 according to the following scales:

[0172] [Tables22]

Quoting Erythema (E) Edema (Oc) 0 no erythema no edema 0.5 Barely noticeable erythema, very slightly pink coloring in part of the patching area barely noticeable palpable edema 1 Light erythema, pink discoloration over the entire patching surface palpable and visible edema 2 Moderate erythema, clear discoloration over the entire patching surface marked edema with or without papules or vesicles 3 Significant erythema, intense coloring over the entire patching area significant edema spreading outside the patching area with or without papules or vesicles

Any other skin reaction (bubbles, papules, vesicles, dryness, scaling, roughness, soap effect, etc.) was evaluated according to the following scale and reported descriptively:

- 0: no reaction,

- 0.5: very light

- 1: light

- 2: moderate

- 3: important.

At the end of the study, an average irritation index (I.I.M.) was calculated according to the following formula:

[0173] [Math. S]

I.LM.= Sum of skin reactions (E + Oe + bubbles + papules + vesicles)/

Number of volunteers analyzed

The I.I.M. obtained made it possible to classify the tested extract according to the scale presented in the table below:

I.I.M. S 0.20 Non-irritating

0.20 < I.I.M. S 0.50 Slightly irritating

0.50 < I.I.M. S 2 Moderately irritating < I.I.M. 3 Very irritating

Results: The Average Irritation Index (I.LM.) of peregrina cake extract is equal to: 0.

Conclusion: Deperegrina oilcake extract can be considered non-irritating after 48 consecutive hours of application in 12 volunteers.

[0174] General conclusion of the tests:

The results of the tests carried out above are conclusive and demonstrate for the peregrina extract according to example 1:

1. Eye and skin irritation tests are negative

2. Phototoxicity tests are negative

3. Mutagenicity tests are negative.

The safety of the peregrina extract according to the invention is demonstrated and ideal for large-scale topical cosmetic use without restriction on target populations.

[0175] Example 18: Evaluation by measuring the Insensible Water Loss (TEWL) on the skin barrier and its acceptability after use over a period of 21 days The product studied in this study is the care product in the form of a cream. example 15. This product is applied morning and evening to the clean face, massaging gently, avoiding the eye area. Measurements are taken at cheek level.

The evaluation criteria are:

- evaluation of the effect on the skin barrier: comparison of TEWL values before any application of the product (Jl) then after 21 days of application (D21).

- increase in possible discomfort sensations on D21.

- cosmetic acceptability: questionnaire completed by the volunteer on D21.

female volunteers aged 50 on average (between 20 and 70) with all skin types were tested. The product evaluated on the skin barrier gave the following results (*=% variation 121 compared to Jl, **=Wilcoxon test for paired data with S= significant (p<0.05 and MS=not significant (p >0.05):

[0176] [Tables23]

TEWL value (g/m ³ .h) Jl D21 Average 11.70 11.79 Standard deviation 3.26 2.87 Median 10.39 11.40 Minimum 7.97 7.12 Maximum 17.30 17.14 % change* - 0.83 p-value** Significance - p=0.555 (NS)

Analysis of the results demonstrates that the TEWL remained stable at D21 compared to Dl: the product presented a “dermo-protective” effect after 21 days of application. Given the absence of a significant drop in TEWL values on D21 compared to Dl, the “nourishing” effect of the product tested could not be demonstrated by instrumental measurements. 81% of volunteers responded positively to the question “the skin is nourished” in the acceptability questionnaire on D21.

Conclusion: Under the conditions of the study, the cream presented a “dermoprotective” effect, evidenced by TEWL measurement and good cosmetic acceptability, with 86% favorable opinions.

Claims (9)

[Claim 1} [Claim 2] [Claim 3] Claims Moringaperegrina seed extract, characterized in that it is obtained by solid-liquid extraction of the cake of unshelled seeds, with stirring, in a proportion of approximately 25% by weight of solid material relative to the total weight used in a predominantly alcoholic solvent, the alcohol being chosen from ethanol or methanol with optionally a co-solvent of the polyol or subcritical water type, in a proportion of 70 to 100% by weight of alcohol relative to the total weight of the solvent, at a temperature between 16 and 30°C for a period of approximately 2 hours and by separation of the liquid and solid phases so as to eliminate the solid phase and recover a liquid extract of Moringa peregrina seed, said extract being rich into the compound 2,5-diformylfuran. Extract according to claim 1, characterized in that the liquid extract obtained is dried so as to obtain a dry extract of the Moringa peregrina seed cake containing more than 50% by weight of 2,5-diformylfuran relative to the total weight of dry matter. Process for obtaining a Moringa peregrina seed extract according to one of claims 1 to 2, characterized in that it comprises the following steps according to which: a) the unshelled seeds of Moringa peregrina are collected and dried to obtain an internal humidity level of less than 8%; b) the dried seeds are pressed so as to separate the oil from the rest of the seed to obtain the cake, c) we grind the cake obtained in step b), d) we disperse the ground obtained in step c) in a proportion of approximately 25% by weight of solid material relative to the total weight used in a predominantly alcoholic solvent, the alcohol being chosen from ethanol or methanol with optionally a co-solvent of the polyol or subcritical water type, in a proportion of 70 to 100% by weight of alcohol relative to the total weight of the solvent; e) a solid-liquid extraction is carried out, with stirring, at a temperature between 16 and 30 ^° C for a period of approximately 2 hours, f) the liquid and solid phases are separated so as to eliminate the solid phase and recover a liquid extract of Moringa peregrina cake, and Ut g) optionally, when the alcohol is ethanol, the extract of Moringa peregrina liquid obtained in such a way as to obtain an extract of Moringa peregrina solid. [Claim 4] Process according to claim 3, characterized in that the majority solvent 5 The purely alcoholic solvent is the pure 96° ethanol solvent. [Claim 5] Process according to claim 3, characterized in that the extract of Moringaperegrina liquid obtained is purified by distillation, microfiltration, ultrafiltration and/or nanofiltration to concentrate the extract in 2,5-diformylfuran compared to organic materials also 10 extracted. [Claim 6] Cosmetic or nutricosmelic composition, characterized in that it comprises, as active agent, an effective quantity of an extract of Moringa peregrina seeds according to one of claims 1 to 2, and a physiologically acceptable excipient. 15 [Claim 7] Composition according to claim 6, characterized in that it is a cosmetic composition formulated to be applied topically to the skin and in that the Moringaperegrina seed extract is present in the composition at a concentration of 0.002 to 20% by weight relative to the total weight of the composition, preferably 20 0.01 to 10%. [Claim 8] Composition according to claim 6, characterized in that it is a nutricosmetic composition formulated to be ingested and in that the Moringa peregrina seed extract is present in the composition at a concentration of 0.01 to 100% by weight per relation to 25 total weight of the composition. [Claim 9] Cosmetic or nutricosmetic use of a composition according to one of claims 6 to 8, to improve the appearance of the skin, mucous membranes or appendages, to relax, soothe and de-stress the skin and prevent and/or fight against skin signs of aging and/or 30 photo-aging and prevent age spots.