Composition comprising an oily macerate of hemp flowers and a Pickering emulsion, preparation and uses

Abstract

Composition comprising a solution containing at least one oily macerate of hemp flowers and an H / W Pickering emulsion containing an aqueous phase, a vegetable oily phase and an organomodified phyllosilicate, process for preparing the composition and use as a soothing, relaxing, anti-stress agent for the skin. Figure to be published with the abstract: none.

Description

Title of the invention: Composition comprising an oily macerate of hemp flowers and a Pickering emulsion, preparation and uses

[0001] The present invention relates to the field of compositions based on hemp flower oil(s). More specifically, the invention relates to a composition comprising at least one oily macerate of hemp flowers as well as its use in cosmetics when applied to the skin, in particular as an anti-stress, soothing, relaxing agent for the skin.

[0002] The skin is the main protective barrier of the human body against external aggressions such as atmospheric pollution, climatic variations and UV radiation.

[0003] The skin is made up of three layers: the epidermis, the dermis and the hypodermis. The epidermis, which is in contact with the superficial layer, is more particularly concerned by interactions with the external environment. The epidermis is covered by a hydrolipidic film and is composed of four layers: the horny layer, the granular layer, the spinous layer and the basal layer.

[0004] Patent application WO2016 / 123475 describes a composition comprising a Cannabis oil extract and vitamin E. In the paragraph devoted to the prior art, the authors highlight the viscous nature of cannabis oils as well as the difficulties which result from this, particularly when one wishes to use them by vaporizing them.

[0005] The viscosity of cannabis oils is therefore a parameter to be taken into consideration when one wishes to develop a composition having a particular galenic adapted to application to the skin. Summary of the invention

[0006] The inventor has shown that a solution to the technical problem of obtaining new soothing cosmetic compositions could be provided by means of a particular composition comprising at least one oily macerate of hemp flowers and a Pickering emulsion.

[0007] Hemp or Cannabis is a flowering herbaceous plant of the Cannabinaceae family. This annual plant, native to Central or South Asia, has several phenotypes that can be described as subspecies and varieties.

[0008] Hemp contains more than 540 active ingredients identified today, belonging to various chemical classes, including cannabinoids, terpenes and flavonoids (1).

[0009] The exploitation of the different parts of the plant makes it possible to meet multiple industrial uses. It is the feminized flowers which are mainly exploited in the medical, well-being and cosmetic sectors, in particular with a view to extracting two molecules from the cannabinoid family: A-9-tetrahydrocannabinol, also called more simply "tetrahydrocannabinol" or THC and Cannabidiol or CBD.

[0010] The scientific community has now established the existence of an "entourage effect" for Cannabis, that is to say an interaction and a synergy of the compounds of interest contained in the plant allowing to arrive at products with an optimized benefit-risk profile (2,3). New attention is therefore today paid to new molecules from the family of terpenes, flavonoids, but also other cannabinoids such as Cannabigerol or CBG.

[0011] The “entourage effect” questions the relevance of historical strategies of development of the aforementioned hemp industries exploiting almost exclusively one or two molecules, THC and CBD, used alone or in combination.

[0012] Surprisingly and advantageously, the inventor has demonstrated a particular composition having advantageous effects in cosmetics when applied to the skin, more particularly as a soothing, relaxing, anti-stress agent.

[0013] The present invention relates to a composition comprising a solution containing at least one oily macerate of hemp flowers and an H / W Pickering emulsion containing an aqueous phase, a vegetable oily phase and an organomodified phyllosilicate.

[0014] Preferably, the solution containing at least one oily macerate of hemp flowers is present in a content ranging from 0.1 to 25%, preferably ranging from 1% to 5% and advantageously 3.4% by weight relative to the total weight of the composition.

[0015] The present invention also relates to a process for preparing the composition according to the invention.

[0016] The present invention also relates to the cosmetic, non-therapeutic use of the composition according to the invention as a soothing, relaxing, anti-stress agent for the skin.

[0017] The present invention also relates to a method for cosmetic, non-therapeutic treatment of the skin, comprising at least one step of applying the composition according to the invention to the skin.

[0018] These cosmetic compositions are furthermore stable, non-irritating, non-toxic, non-allergenic to the skin.

[0019] Other aspects, advantages, properties of the present invention are presented in the description and examples which follow. Detailed description

[0020] Definitions

[0021] In this text, unless specifically indicated otherwise, the percentages are expressed by weight of a reference composition.

[0022] In this text, intervals are defined in an abbreviated manner in order to avoid describing each and every value in that interval, however any suitable value in the interval may be chosen as the upper value, the lower value or the terminal values ​​of the interval. For example, an interval of 0.1 to 1.0 represents the terminal values ​​of 0.1 and 1.0, as well as the intermediate values ​​of 0.2; 0.3; 0.4; 0.5; 0.6; 0.7; 0.8; 0.9 and all intermediate intervals within 0.1 to 1.0, such as 0.2 to 0.5; 0.2 to 0.8; 0.7 to 1.0...Unless otherwise stated, an interval defined as "between value A and value B" includes values ​​A and B and is therefore equivalent to an interval "from value A to value B", the expression "at least" includes the value stated after, for example, "at least 5%" should be understood as also including "5%", the expression "a maximum of" includes the value stated after, for example, "a maximum of 5%" should be understood as also including "5%".

[0023] Furthermore, in the present text, measurable values, such as a quantity, are to be understood as including standard deviations that can be readily determined by a person skilled in the art in the reference technical field. Preferably, these values ​​are intended to include variations of ± 5%.

[0024] In this text, the words “hemp” and “Cannabis” are used equivalently.

[0025] By “skin” we mean the epidermis of the face or body or the scalp.

[0026] The compositions according to the present invention are cosmetic, non-therapeutic compositions. The cosmetic compositions are intended to be applied to healthy skin, by simple massage to treat the epidermis. They comply with EC Regulation 1223 / 2009.

[0027] Hemp oil macerate(s)

[0028] The solution containing at least one oily macerate of hemp flowers which can be used according to the invention is a solution comprising at least 50%, preferably at least 75% by weight of an oily macerate obtained by maceration of hemp flowers in a hemp seed oil.

[0029] Preferably this macerate is obtained by maceration of 500 to 1000g of hemp flowers for 1 L of hemp seed oil.

[0030] The ratio of mass of hemp flowers / hemp seed oil is advantageously between 1 / 3 and 3 / 1 and preferably between 1 / 2 and 1 / 1.

[0031] In general, the duration of the maceration, also called "infusion" is 3 to 4 weeks. It is within the skill of the person skilled in the art to define the duration of the maceration.

[0032] All hemp flowers are usable, provided, of course, that their use is authorized by the national or transnational regulations in question.

[0033] In Europe, this is particularly the case for strains with a high CBD content from the Common Catalogue of Varieties of Agricultural Plant Species, such as Kompolti, Carmagola, Perugina, Finola, or more recently Pain Killer, Midwest or Strawberry K. In other countries, particularly in the United States, other genetic strains of interest can be used, such as, for example, ACDC, Charlotte's web, Harlequin, Harle-Tsu, Ringo's gift.

[0034] Advantageously, the flowers used are manicured, that is to say that the inflorescence has been harvested by hand in order to preserve as much as possible the quality of the flower totum.

[0035] To this main macerate can be added another, secondary macerate of hemp flowers in a vegetable oil other than hemp seed oil, for example sunflower oil.

[0036] The addition of a second macerate in which other hemp flowers have been decarboxylated makes it possible to guarantee a better “entourage effect” of the solution according to the invention, that is to say to guarantee that a greater variety of compounds from hemp flowers, with their specific cosmetic properties, are present in the solution.

[0037] Decarboxylation consists of heating an oily macerate to convert the cannabinoids from their acid form to their active form; decarboxylation can also be achieved by the action of UV on the macerate. This process converts, in particular, CBDA (Cannabidiol Acid) into CBD and THCA (Tetrahydrocannabinol Acid) into THC.

[0038] Advantageously, the volume ratio (v / v) between the oily macerate obtained by maceration of hemp flowers in a hemp seed oil (main macerate) and the oily macerate obtained by maceration of hemp flowers in a different oil (secondary macerate) is strictly greater than 1 / 1, preferably it is 3 / 1.

[0039] Preferably, the solution containing at least one oily macerate of hemp flowers comprises at least CBD in an amount ranging from 0.005% to 24.000%, preferably from 0.010% to 5.000% by weight relative to the total weight of the solution; CBDA in an amount ranging from 0.005% to 24.000%, preferably 0.010% to 5.000% by weight relative to the total weight of the solution; THC in an amount ranging from 0.001% to 30.000%, preferably 0.010% to 1.000% by weight relative to the total weight of the solution; CBG in an amount ranging from 0.001% to 22.000%, preferably 0.010% to 3.000% by weight relative to the total weight of the solution and CBGA in an amount ranging from 0.001% to 22.000%, preferably 0.010% to 3.000% by weight relative to the total weight of the solution.

[0040] Pickering Emulsions

[0041] Pickering emulsions are emulsions stabilized by solid particles. During the preparation of the emulsion, said solid particles are positioned at the interface between the aqueous phase and the oily phase.

[0042] The Pickering emulsions according to the present invention are oil-in-water emulsions, i.e. O / W, stabilized by a particular clay which is a modified natural phyllosilicate.

[0043] Advantageously, the aqueous phase of the Pickering emulsion is present in an amount ranging from 51% to 90%, preferably 55% to 70% and more preferably 58% to 65% by weight relative to the total weight of the composition.

[0044] According to a preferred embodiment, the modified natural phyllosilicate comprises a phyllosilicate selected from the group consisting of vermiculites and smectites. Preferably, the phyllosilicate may be selected from the group consisting of montmorillonites, bentonites, nonytronites, beidellites, volkonskoites, hectorites, saponites, sauconites, sobockites, stevensites, svinfordites, preferably these are phyllosilicates of sodium, potassium, calcium, or mixtures thereof. More preferably, the phyllosilicate is selected from the group consisting of hectorite, montmorillonite, bentonite or mixtures thereof.

[0045] According to a preferred embodiment, the modified natural phyllosilicate comprises an organic compound selected from the group consisting of xanthan gum, guar gum, Tara or Carob gum, Acacia gum, carrageenan, alginate, chitosan, pectin, citric acid, tartaric acid, oxalic acid, succinic acid, malic acid, acetic acid, lactic acid, propionic acid, salicylic acid, glycosaminoglycans. Advantageously, the modified natural phyllosilicate comprises a bentonite modified with xanthan gum and citric acid.

[0046] According to another embodiment, the modified natural phyllosilicate comprises an organic compound chosen from any organic compound known in the art of Pickering emulsions, such as those disclosed in French patent No. FR 2,976,503.

[0047] Advantageously, the modified phyllosilicate is present in an amount ranging from 3% to 20%, preferably 4% to 10% and more preferably 5% to 8% by weight relative to the total weight of the composition.

[0048] According to a preferred embodiment, the vegetable oily phase of the Pickering emulsion comprises at least one vegetable oil chosen from the group formed by hemp oil, sunflower oil, rapeseed oil, olive oil, camelina oil, peanut oil, coconut oil, grape seed oil, castor oil, argan oil, Djansang oil, desert date oil, black cumin oil, prickly pear oil, macadamia oil, soybean oil, palm and palm oil, Tamanu oil, sesame oil, linseed oil, walnut oil, hazelnut oil, baobab oil, passion fruit oil, Brazil nut oil, hibiscus oil, Pumpkin seed, Luffa oil, Carapa oil, Evening Primrose oil, Borage oil, Avocado oil, Almond oil, Sea buckthorn oil, Apricot kernel oil, Cherry kernel oil, Apple seed oil, Pomegranate oil,Jojoba oil, Rosehip oil, Plum oil, Shea butter, Cocoa butter, Kokum butter, Mango butter, Moabi butter, Karanja butter, Tucuma butter, Cupuaçu butter, Buriti butter, Murumuru butter, Kombo butter, Kpangnan butter, Caprylic / capric acid triglycerides.

[0049] Advantageously, the oily phase of the Pickering emulsion is present in an amount ranging from 5% to 40%, preferably 10% to 30% and more preferably 15% to 25% by weight relative to the total weight of the composition.

[0050] Composition

[0051] Advantageously, the composition according to the present invention comprises, in an acceptable medium, at least one cosmetic agent chosen from humectants, thickeners, texturizing agents, emulsifiers, dispersing agents, foaming agents, emulients, preservatives, colorants, plant extracts, plant fibers, minerals, pH correcting agents, active ingredients and perfumes.

[0052] Advantageously, the composition according to the present invention comprises from 0.001 to 20% by weight of at least one cosmetic agent relative to the total weight of the composition.

[0053] Of course, those skilled in the art will take care to choose these cosmetic agents so as not to alter the properties of the composition according to the invention.

[0054] The composition according to the invention may constitute a massage composition, a skin care composition, and in particular a cleansing, protective, treatment or care cream for the face, for the hands, for the feet, or for the body, in particular the composition may be a day cream, night cream, make-up remover cream, cream foundation, sunscreen; a fluid foundation, a cleansing milk, a protective or care body milk, an anti-sun milk; a lotion, gel or mousse for skin care, such as a micellar cleansing lotion.

[0055] Advantageously, the process for preparing the composition according to the invention comprises at least the following steps in this order: preparing the aqueous phase; add the organomodified phyllosilicate to the aqueous phase and mix; add the vegetable oil phase to the mixture obtained and obtain a Pickering H / W emulsion; add the solution containing at least one oily macerate of hemp flowers to the Pickering H / W emulsion.

[0056] Of course, the vegetable oil phase of the Pickering emulsion is different from the solution containing at least one oily macerate of hemp flowers.

[0057] It has been noted that the viscosities of the oily macerates of hemp flowers are compatible with the process for preparing the compositions according to the invention.

[0058] Uses

[0059] The cosmetic composition according to the invention is advantageously intended to be applied to the skin.

[0060] The present invention also relates to the cosmetic, non-therapeutic use of the composition according to the invention as a soothing, relaxing, anti-stress agent for the skin. It also relates to a cosmetic, non-therapeutic treatment method for the skin, comprising at least one step of applying the composition according to the invention to the skin.

[0061] The following examples are intended to illustrate the invention without limiting its scope. Examples

[0062] A. Macerates

[0063] Example A 1- my cerate 1

[0064] Table 1 lists the products used to prepare the macerates

[0065] [Tables 1] Product Quantity Supplier Organic hemp seed oil 1st cold pressed 7.5 L AB CD (France) Deflavored sunflower oil 2.5 L (Bouton D'or Bio) France Kompolti Hemp - Manicured flowers 5000 g AB CD (France) Carmagnola hemp - Manicured flowers 2000 g AB CD (France) Perugina hemp - Manicured flowers (under greenhouse) 1000 g AB CD (France)

[0066] The hemp flowers used are all dried flowers manicured then crushed with a cut-off threshold of 250 microns.

[0067] They are then mixed to form a mix of crushed hemp flowers.

[0068] Preparation

[0069] In a first reactor were introduced 2.5 L of dearomatized sunflower oil and 2000 g of crushed flower mix. This mixture is then heated in a water bath, at 95 °C, for 1 hour for the decarboxylation of certain active components (transformation of CBA into CBD, CBGA into CBG and THCA into THC).

[0070] This “decarboxylated” oil is then filtered.

[0071] In a second reactor were introduced the 7.5 L of hemp seed oil and 6000 g of crushed flower mix. This mixture is then left to macerate for 3 weeks. During these 3 weeks, the macerate is stirred every day for 3 minutes.

[0072] The macerate obtained in the second reactor is then left to settle for 3 days, i.e. it is not stirred. It is then filtered using a coffee filter. The flowers remaining at the bottom of the settling tank are recovered and pressed using an ECOLEA oil press.

[0073] The decantation oil, the bottom pressing oil - from the second reactor - and the sunflower oil - from the first reactor - are combined. The oil obtained is called Macerate 1.

[0074] HPLC analysis, with a Shimadzu DAD (diode array detector), of macerate 1 made it possible to determine the following contents recorded in Table 2:

[0075] [Tables 2] Compounds % mass CBD 0.188 CBDA 0.543 THC 0.024 CBG 0.096 CBGA 0.553

[0076] This gives a total CBD content of 0.664% and a total CBG content of 0.582%. To obtain these values, the acid forms (CBD A and CB GA) were multiplied by a factor of 0.878.

[0077] Example A 2- macerate 2

[0078] Table 3 lists the products used to prepare the macerates.

[0079] [Tables3] Product Quantity Supplier Organic Hemp Seed Oil 1st cold pressed 7.5 L AB CD (France) Deflavored sunflower oil 2.5 L (Bouton D'or Bio) France Kompolti Hemp - Manicured flowers 3000 g AB CD (France) Carmagnola Hemp - Manicured flowers 1000 g AB CD (France) Perugina Hemp - Manicured flowers (in greenhouse) 1000 g AB CD (France) Kompolti Trichome (powder from flowers crushed to 250 pm in olive oil) 500 g AB CD (France)

[0080] The hemp flowers used are all dried flowers manicured then crushed with a cut-off threshold of 250 microns.

[0081] They are then mixed to form a mix of crushed hemp flowers.

[0082] Preparation

[0083] In a first reactor were introduced 2.5 L of dearomatized sunflower oil and 1250 g of crushed flower mix. This mixture was then heated over low heat, at 125°C, for 30 minutes in order to decarboxylate certain active components (transformation of CBA into CBD, CBGA into CBG and THCA into THC). This decarboxylated oil is then filtered.

[0084] In a second reactor were introduced the 7.5 L of hemp oil, 3750 g of crushed flower mix and 500 g of Kompolti Trichome. This mixture is then left to macerate for 3 weeks. During these 3 weeks, the macerate is stirred every day for 3 minutes.

[0085] The macerate obtained in the second reactor is then left to settle for 3 days, i.e. it is not stirred. It is then filtered using a coffee filter.

[0086] The decantation oil - from the second reactor - and the sunflower oil - from the first reactor - are combined. The oil obtained is called Macerate 2.

[0087] HPLC analysis, with a Thermo Scientific Vanquish DAD, of macerate 2 made it possible to determine the following contents recorded in Table 4:

[0088] [Tables4] Compounds % mass CBD 2.025 CBDA 0.596 THC 0.032 CBG 0.029 CBGA 0.239

[0089] This gives a total CBD content of 2.548% and a total CBG content of 0.239%. To obtain these values, the acid forms (CBD A and CB GA) were multiplied by a factor of 0.878.

[0090] Example A 3- macerate 3

[0091] Table 3 lists the products used to prepare the macerates.

[0092] [Tables5] Product Quantity Supplier Organic Hemp Seed Oil 1st Cold Pressed 10.0 L AB CD (France) Kompolti Hemp - Manicured Flowers 5000 g AB CD (France) Carmagnola Hemp - Manicured Flowers 1000 g AB CD (France)

[0093] The hemp flowers used are all dried flowers manicured then crushed with a cut-off threshold of 250 microns.

[0094] 2500 g of Kompolti flowers and 1000 g of Carmagnola flowers are crushed and mixed to form a mix of crushed hemp flowers.

[0095] Preparation

[0096] The remaining 2500 g of Kompolti flowers are heated in the oven at 120°C for 20 minutes. The treatment leads to decarboxylation of the Kompolti flowers.

[0097] 10.0 L of hemp oil, 2500 g of decarboxylated Kompolti flowers and 3500 g of crushed hemp flower mix were introduced into a reactor. This mixture was then left to macerate for 3 weeks. During these 3 weeks, the macerate was stirred every day for 3 minutes.

[0098] The macerate obtained is then filtered using a coffee filter.

[0099] The oil obtained is called Macerate 3.

[0100] HPLC analysis, with a Thermo Scientific Vanquish DAD, of macerate 3 made it possible to determine the following contents recorded in Table 6.

[0101] [Tableauxô] Compounds % mass CBD 1.044 CBDA 0.521 THC 0.033 CBG 0.030 CBGA 0.084

[0102] This gives a total CBD content of 1.502% and a total CBG content of 0.103%. To obtain these values, the acid forms (CBD A and CB GA) were multiplied by a factor of 0.878.

[0103] It has been verified that macerates 1, 2 and 3 are similar with respect to the intended soothing properties.

[0104] B. Compositions tested

[0105] 1- Comparative composition Cl

[0106] The comparative composition Cl corresponds to the macerate ML

[0107] 2- Comparative composition C2

[0108] Comparative composition C 2 comprises a Pickering H / W emulsion containing a clay which is a modified phyllosilicate (without hemp oil macerate).

[0109] Table 7 lists the products used to prepare composition C2.

[0110] [Tables7] Phase Trade name % INCI A Water 73.70 Aqua B Georgard Ultra marketed by Lo nza 1.00 Gluconolactone & Sodium benzoate & Calcium gluconate B Sodium benzoate 0.30 Sodium benzoate C Frametime CXG commercialized by E phyla 4.50 Bentonite & xanthan gum & Sodium ste aroyl glutamate & citric acid

[0111] Preparation of composition C2:

[0112] Phases A and B are mixed to form an aqueous phase, then phase C is added and everything is mixed. Finally, the oily phase D is added and everything is mixed. Once the Pickering emulsion is obtained, the pH is adjusted to 5.00.

[0113] 3-Composition C3

[0114] Comparative composition C 3 comprises a macerate and an H / W Pickering emulsion containing a kaolin.

[0115] Table 8 lists the products used to prepare composition C3.

[0116] [Tables8] Phase Commercial name % INCI A Water 70.30 Aqua B Georgard Ultra marketed by Lo nza 1.00 Gluconolactone & Sodium benzoate & Calcium gluconate B Sodiu m benzoate 0.30 Sodium benzoate C White clay marketed by CO OPER 4.50 Kaolin C Xanthan gum FF commercial Used by Jungbunz lauer 0.50 xanthan gum D Commercial refined sunflower oil produced by Cauvin 20.00 Helianthus annuus seed oil E Macerate 1 3.40 Total 100

[0117] Preparation of composition C3:

[0118] Phases A and B are mixed to form an aqueous phase, then phase C is added and everything is mixed. Finally, oily phase D is added and everything is mixed.

[0119] Once the Pickering emulsion is obtained, phase E, i.e. macerate 1, is added and everything is mixed. Finally, the pH is adjusted to 5.00.

[0120] 4- Composition II in accordance with the invention

[0121] Composition II comprises a macerate and an H / W Pickering emulsion containing a clay which is a modified phyllosilicate.

[0122] Table 9 lists the products used to prepare composition II.

[0123] [Tables9] Phase Commercial name % INCI A Water qsp Aqua B Frametime CX commercialized by Ep hyla 6.00 Bentonite & xanthan gum & citric acid B Amisoft Hsllp 0.30 Sodium stearoyl glutamate B Xanthan gum FF commercialized Used by Jungbunz lauer 0.50 Xanthan gum C Cetosrearyl alcohol 50 / 50 4.00 Cetearyl alcohol C Ephyster MCR 20.00 Brassica napus extract C HTR1 marketed by EPHYLA 1.50 Helianthus annuus seed oil & Protium h eptaphyllum resin C Sepicide LD 1.00 Phenoxyethanol D Macerate 1 3.40 total 100.00

[0124] Preparation of composition II

[0125] Phases A and B are mixed to form an aqueous phase, then phase C is added and everything is mixed.

[0126] Once the Pickering emulsion is obtained, phase D, i.e. macerate 1, is added and everything is mixed. Finally, the pH is adjusted to 5.00.

[0127] C. Activity on a skin enzyme in an in vitro model

[0128] Tests were carried out in vitro on the key enzyme of the arachidonic cascade: the lipoxygenase, an enzyme present in the epidermis of the skin.

[0129] This study is based on the influence of cosmetic products on the activity of the Lipoxygenase enzyme so as to be able to compare and classify the influence of these products on this enzymatic activity. This study aims to illustrate the impact of topical galenics on an enzymatic activity of the skin.

[0130] Measurements of the lipoxygenase enzyme activity of several compositions were carried out in an in vitro cellular model using a kit.

[0131] Materials and Reagents

[0132] The analysis kit used is the kit sold under the name “Lipoxygenase Inhibitor Screening Assay Kit” by the company CAYMAN / INTERCHIM.

[0133] Table 10 lists the components of the kit.

[0134] [TableauxlO] Concentrated TRIS-HCL BUFFER CAYMAN / INTERCHIM CHROMOGEN 1 = Development Reagent 1 CAYMAN / INTERCHIM CHROMOGEN 2 = Development Reagent 2 CAYMAN / INTERCHIM ENZYME: 15-LIPOXYGENASE CAYMAN / INTERCHIM ARACHIDONIC ACID CAYMAN / INTERCHIM POTASSIUM HYDROXIDE 0.1 M CAYMAN / INTERCHIM NDGA INHIBITOR CAYMAN / INTERCHIM

[0135] Spectrometer

[0136] The measurements were carried out using a POLARstar Omega spectrometer from BMG LABTECH.

[0137] Preparation of reagents

[0138] 1. RI: Lipoxygenase Inhibitor Screening Test Buffer - Dilute 3 ml of concentrated BUFFER with 27 ml of HPLC grade water. RI Test Buffer (0.1 M Tris-HCl, pH 7.4) should be used for dilution of samples and ENZYME: 15-LIPOXYGENASE (15-LO standard) prior to assay. When stored at 4°C, this RI Test Buffer is stable for at least two months.

[0139] R2 Chromogen Prepare Chromogen R2 before use by mixing equal volumes of CHROMOGEN 1 and CHROMOGEN 2 in a test tube and vortexing. The volume of Chromogen R2 to be prepared depends on the number of wells analyzed (100 μl for each well). Chromogen R2 should be used within one hour.

[0140] R3: 15-Lipoxygenase Standard -

[0141] Transfer 10 μl of ENZYME: 15-LIPOXYGENASE into a vial and dilute with 990 μl of RI before use, keep on ice and use within one hour.

[0142] R4 Arachidonic acid (substrate) -

[0143] The bottle provided in the kit contains a solution of arachidonic acid (ARACHIDONIC ACID) in ethanol and should be stored at -20°C. Transfer 25 µl of the ARACHIDONIC ACID product to another vial, add 25 µl of the potassium hydroxide solution, vortex, and dilute with 950 µl of HPLC-grade water to obtain the 1 mM R4 solution. A 10 µl aliquot will give a reaction concentration of 91 pM in the wells.

[0144] The kit contains one vial of ready-to-use 0.1 M potassium hydroxide (KOH)

[0145] R5 NDGA Positive Control Inhibitor - The kit contains one vial of 550 nmol of the non-selective lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA). Resuspend in 500 µl of RI to make a 1.1 mm stock. Adding 10 µl to the test gives a final concentration of 100 pM inhibitor in the well.

[0146] • Each well has a final volume of 210 μl.

[0147] Test execution

[0148] 1. Blank Wells Samples (Blank Sample): add 90 µl of RI solution and 10 µl of the test product in triplicate

[0149] 2. Positive control well (basal activity = Control +):

[0150] add 90 μl of solution R3 and 10 μl of solution RI in triplicate.

[0151] 3. Well Product under test:

[0152] add 90 μl of solution R3 and 10 μl of the test product in triplicate.

[0153] 4. Inhibitor well (Witness -)

[0154] add 90 μl of solution R3, 10 μl of solution R5 in triplicate;

[0155] 5. Incubate for five minutes at room temperature.

[0156] 6. Initiate the reaction by adding 10 µl of solution R4 to all wells. Place the 96-well plate on a shaker for at least ten minutes.

[0157] 7. Add 100 µl of solution R2 to each well to develop the reaction. Cover with a plate lid and place the 96-well plate on a shaker for five minutes.

[0158] 8. Remove the lid and read the absorbance at 500 nm using a plate reader.

[0159] Principle of the reaction Test system

[0160] A buffered solution of lipoxygenase (R3) reacts with a specific substrate, arachidonic acid (R4), and transforms it to form a compound that binds to a chromogen (R2), under stirring at room temperature. The activity of the lipoxygenase can thus be evaluated by measuring the absorbance at 500 nm.

[0161] The sample or reference product: NDGA inhibitor (R5) is brought into contact with the lipoxygenase solution (R3) at the same time as the enzyme substrate (R4). The substrate (R4) transformed by the enzyme is colored using the chromogen R2 by stirring at room temperature. The lipoxygenase activity in the different wells is then evaluated by measuring the absorbance at 500 nm.

[0162] The modulation of this activity is expressed as a percentage of inhibition or activation of the lipoxygenase activity compared to the basal level of activity in the absence of sample or reference product, i.e. only in the presence of the enzyme substrate (arachidonic acid).

[0163] Incubation protocol

[0164] A solution of lipoxygenase enzyme is incubated in its substrate, arachidonic acid, for 10 minutes, in the absence or presence of the reference inhibitor and the sample tested, then the chromogen is incorporated before a 35-minute incubation at room temperature on an orbital shaker of the Incu-Shaker Mini type from the company Benchmark.

[0165] Evaluation of effects

[0166] At the end of the incubation period, the activity of the lipoxygenase enzyme with and without test or reference product was evaluated by measuring the absorbance of the reaction media at 500 nm.

[0167] For each concentration tested, the modulation of the activity of the lipoxygenase enzyme by the product under test is calculated according to the following formula

[0168] Percentage modulation of lipoxygenase enzyme activity =

[0169] 100 x [(DO test product - DO lipoxygenase alone (blank)) / DO lipoxygenase alone]

[0170] If the result is negative, the percentage is expressed as inhibition of the enzyme.

[0171] The results in terms of mean OD, mean OD minus blank, % enzyme activity are presented in Table 11.

[0172] [Tableauxll] Sample Average D Sample Average DO Sample DO blank Enzyme activity (%) Blank 0.221 0.000 0.00 T+ 0.436 0.215 100.00 T- 0.187 -0.034 0.00 Cl 0.812 0.217 101.24 C2 1.653 -0.073 0.00 C3 3.845 0.054 25.00 II 2.437 0.578 269.10

[0173] The level of significance between the "control" and the "reference product" was assessed using a Student's t-test (*: p<0.05).

[0174] The level of significance between the "control" and each "tested compound" was assessed independently for each composition by a one-way analysis of variance (one-way ANOVA) followed by a Holm-Sidak test (*: p<0.05).

[0175] Conclusion s

[0176] The composition according to the invention II has an enzymatic activity of 269% (i.e. +169% compared to the normal basal activity of 100%) whereas the comparative composition Cl of hemp oily macerate alone has an enzymatic activity of 101% and the comparative composition C 2 comprising the Pickering emulsion alone has an enzymatic activity of 0%.

[0177] It should also be noted that the comparative composition C3 comprising the hemp oil macerate and a kaolin-based Pickering emulsion has an enzymatic activity of only 25%.

[0178] The composition according to the invention exhibits a synergistic effect on the enzymatic activity studied which is not observed when a kaolin-based Pickering emulsion is used.

[0179] Surprisingly and unexpectedly, the composition according to the invention II strongly stimulates the enzymatic activity, whereas the composition Cl, i.e. macerate 1, does not show any modulation of the basal rate, and the composition C2 (which constitutes the skeleton of the composition according to the invention) without macerate 1, on the contrary, totally inhibits the enzymatic activity. Also surprisingly, the composition C3 comprising a Pickering emulsion made from kaolin shows an inhibition of the order of 75% of the enzymatic activity compared to the basal (= normal) rate of the enzyme activity.

[0180] Macerate 1 (Cl) alone does not modulate the basal activity of the enzyme, while the pickering galenic (C2) provides an inhibitory effect on the enzymatic activity, and on the contrary the composition according to the invention (II) provides stimulation.

[0181] D- Measurement of the relaxing or de-stressing effect on human skin explants by measuring IL-6 (secretion of interleukin 6).

[0182] The skin, particularly the epidermis, secretes a basal level of cellular mediator which reflects both intercellular stress and environmental stress (it is a form of expression of the reactivity of our epidermis in contact with its external environment).

[0183] This study is based on the influence of the composition in accordance with the invention II on the capacity to modulate Interleukin 6 production.

[0184] The desired effect is an inhibitory effect which results in a soothing or relaxing or anti-stress effect on the epidermis which is in contact with the external environment.

[0185] Principle of the study

[0186] For this study, normal fresh human skin explants are exposed to the test samples for a contact time of 6 hours.

[0187] At the end of this contact time, the explants were dissected and suspended in physiological serum, the IL-6 concentration of the explants is determined by Elisa assay using an Enzo IL-6 kit.

[0188] Test system

[0189] For this study, human skin explants are exposed to the test samples before being placed on Franz cells; placement on Franz cells induces expected environmental stress because the skin is pinched and / or clipped onto the edges of the Franz cell.

[0190] The skin explant / Franz cell systems are maintained for 6 hours at 32°C ± 1°C. At the end of the incubation, the explants are rinsed, dissected and stored in physiological saline before being subjected to an ultrasound cycle for 30 minutes. The explants are then centrifuged for 15 minutes at 5000 rpm and at 4°C, then the supernatants are preserved and used for the determination of interleukins 6.

[0191] Skin explant supernatants are used for the measurement of interleukins 6 using the Enzo IL-6 (human) high sensitivity ELISA Kit. This kit provides a practical test for the measurement of interleukins 6, thus making it possible to evaluate the modulating effect of active ingredients or topical products to be tested. This kit implements an immuno-enzymatic assay method for which the samples are brought into contact with monoclonal antibodies specific to interleukins 6 (IL-6) fixed on a microplate. After rinsing, a biotinylated anti-interleukin-6 antibody is added and binds specifically. A peroxidase solution is added, binds to the biotinylated antibodies and then reacts with the addition of a substrate to form a blue color. The reaction is stopped and a yellow color is formed, the intensity of which is proportional to the concentration of interleukins 6. At the same time, an IL-6 calibration curve is produced and will be used to determine the IL-6 concentration in the supernatants of skin explants.

[0192] In this study, each condition and / or sample tested was analyzed in triplicate with the explants and for the IL-6 assay.

[0193] Results

[0194] Standard range

[0195] For this study, a calibration range of interleukins 6 ranging from 0.0 to 50.0 pg / mL was carried out. The absorbances of the standard range of interleukins 6 at 450 nm are presented in Table 12.

[0196] [Tables 12] Interleukin 6 range (pg / mL) White NSB 0.0 0.5 1.0 5.0 10.0 50.0 OD 450 nm 0.000 0.096 0.572 0.107 0.266 1.012 0.949 2.377 OD 450 nm-NS B - 0.000 0.476 0.01 0.170 0.916 0.853 2.281

[0197] NSB: physiological serum.

[0198] Between 0.5 and 5.0 pg / mL of IL-6, the calibration curve is linear. The equation of this line a corresponds to y = 0.1954x-0.0578

[0199] Sample measurement

[0200] To carry out this study, two controls were produced: a “basal” control for the determination of basal stress and a solvent control, representative of the stress caused by the method and the solvent used.

[0201] The absorbance (OD) results at 450 nm of the controls and samples are presented in Tables 13 and 14, below, respectively.

[0202] [Tables 13] OD 450 nm of controls Basal control Solvent control Replicate 1 Replicate 2 Replicate 3 Replicate 1 Replicate 2 Replicate 3 OD 450nm 0.711 1.377 1.086 0.306 0.710 0.447 OD 450nm - NSB 0.615 1.281 0.990 0.210 0.614 0.351

[0203] [Tablesl4] OD 450nm of composition II samples Replicate 1 Replicate 2 OD 450nm 0.712 0.708 OD 450nm -NSB 0.616 0.612

[0204] IL-6 concentrations

[0205] The IL-6 concentrations of the controls and samples were determined using the calibration curve equation and are presented in Tables 15 and 16, respectively, below:

[0206] [Tables 15] OD 450 nm of controls Basal control Solvent control Replicate 1 Replicate 2 Replicate 3 Replicate 1 Replicate 2 Replicate 3 IL-6 concentration (pg / mL) 3.44* 6.85 5.36 1.37 3.44* 2.09 Mean 6.106 1.729 Standard deviation 1.055 0.510

[0207] * values ​​excluded because statistically aberrant, not interpretable

[0208] [Tables 16] OD 450nm of composition II samples Replicate 1 Replicate 2 IL-6 concentration (pg / mL) 3.45 3.43 Mean 3.438 Standard deviation 0.014

[0209] Under the study conditions, the IL-6 concentration of the basal stress control is 6.106 pg / mL and of the solvent-induced stress control 1.729 pg / mL.

[0210] The average IL-6 concentration of the explants exposed to the composition according to the invention II (0.008g) is 3.438 pg / mL, i.e. 43.7% inhibition of the response compared to the basal control.

[0211] Conclusion:

[0212] The explants exposed to the sample “Composition according to the invention II” have an inhibited response compared to the basal control; at the dose of 0.008 g the tested product inhibits the IL-6 response by 43.7%; the “Composition according to the invention II” has a very significant soothing, relaxing or anti-stress effect on this skin explant and more particularly on the epidermis exposed to environmental stress.

[0213] BIBLIOGRAPHY 1. André CM, Hausman JF, Guerriero G. Cannabis sativa: The plant of the thousand and one molecules. Front Plant Sci. 2016; 7:19. 2. LaVigne JE, Hecksel R, Keresztes A, Streicher JM. Cannabis Sativa terpenes are cannbimetic and selectively enhance cannabinoid activity. Nature Scientific Reports. 2021; 11:83232. 3. Russo EB. The Case for the Entourage Effect and Conventional Breeding of Clinical Cannabis: No “Strain,” No Gain. Frontiers in Plant Science. Perspective. 2019; 9:1-8.

Claims

Claims

1. Composition comprising a solution containing at least one oily macerate of hemp flowers and an H / W Pickering emulsion containing an aqueous phase, a vegetable oily phase and an organomodified phyllosilicate.

2. Composition according to claim 1 wherein the solution containing at least one oily macerate of hemp flowers comprises at least CBD in an amount ranging from 0.005% to 24.000%, preferably from 0.01% to 5.00% by weight relative to the total weight of the solution; CBDA in an amount ranging from 0.005% to 24.000%, preferably 0.010% to 5.000% by weight relative to the total weight of the solution; THC in an amount ranging from 0.001% to 30.000%, preferably 0.010% to 1.000% by weight relative to the total weight of the solution; CBG in an amount ranging from 0.001% to 22.000%, preferably 0.010% to 3.000% by weight relative to the total weight of the solution and CB GA in an amount ranging from 0.001% to 22.000%, preferably 0.010% to 3.000% by weight relative to the total weight of the solution.

3. Composition according to any one of claims 1 or 2, characterized in that the solution containing at least one oily macerate of hemp flowers is present in a content ranging from 0.1 to 25%, preferably ranging from 1% to 5% and advantageously 3.4% by weight relative to the total weight of the composition.

4. Composition according to any one of claims 1 or 2 in which the modified phyllosilicate is present in an amount ranging from 3% to 20%, preferably 4% to 10% and more preferably 5% to 8% by weight relative to the total weight of the composition.

5. Composition according to any one of claims 1 to 3 in which the aqueous phase of the Pickering emulsion is present in an amount ranging from 51% to 90%, preferably 55% to 70% and more preferably 58% to 65% by weight relative to the total weight of the composition.

6. Composition according to any one of claims 1 to 4 in which the vegetable oily phase of the Pickering emulsion comprises at least one vegetable oil chosen from the group formed by hemp oil, sunflower oil, rapeseed oil, olive oil, camelina oil, peanut oil, coconut oil, Grape seed oil, Castor oil, Argan oil, Djansang oil, Desert date oil, Black cumin oil, Prickly pear oil, Macadamia oil, Soybean oil, Palm and palm oil, Tamanu oil, Sesame oil, Linseed oil, Walnut oil, Hazelnut oil, Baobab oil, Passion fruit oil, Brazil nut oil, Hibiscus oil, Pumpkin seed oil, Luffa oil, Carapa oil, Evening Primrose oil, Borage oil, Avocado oil, Almond oil, Sea buckthorn oil, Apricot kernel oil, Cherry kernel oil, Apple seed, Pomegranate oil, Jojoba oil, Rosehip oil, Plum oil, Shea butter, Cocoa butter, Kokum butter, Mango butter, Moabi butter, Karanja butter, Tucuma butter, Cupuaçu butter, Buriti butter, Murumuru butter, Kombo butter,Kpangnan butter, caprylic / capric acid triglycerides.,

7. Composition according to any one of claims 1 to 5 in which the oily phase of the Pickering emulsion is present in an amount ranging from 5% to 40%, preferably 10% to 30% and more preferably 15% to 25% by weight relative to the total weight of the composition.

8. Composition according to any one of claims 1 to 6 comprising, in an acceptable medium, at least one cosmetic agent chosen from humectants, thickeners, texturizing agents, emulsifiers, dispersing agents, foaming agents, emulients, preservatives, colorants, plant extracts, plant fibers, minerals, pH correcting agents, active ingredients and perfumes.

9. Composition according to any one of claims 1 to 8, characterized in that it is intended to be applied to the skin.

10. A process for preparing the composition according to any one of claims 1 to 9 comprising at least the following steps in this order: preparing the aqueous phase; adding the organomodified phyllosilicate to the aqueous phase and mixing; adding the vegetable oil phase to the mixture obtained and obtaining an H / W Pickering emulsion; 24 add the solution containing at least one oily macerate of hemp flowers to the Pickering H / W emulsion.

11. Cosmetic, non-therapeutic use of the composition according to any one of claims 1 to 9 as a soothing, relaxing, anti-stress agent for the skin.

12. Method for cosmetic, non-therapeutic treatment of the skin, comprising at least one step of applying to the skin the composition according to any one of claims 1 to 9.