Cosmetic use of compounds derived from oleanolic acid

Abstract

The invention relates to the cosmetic and non-therapeutic use of a composition consisting essentially of a compound of formula I below: (I) or its enantiomers, isomers or diastereoisomers, or a mixture thereof, or a salt or a solvate thereof, wherein R1 and R2, independently of each other, are a C3-C6 sugar, a uronic acid, or a glucoside, for bleaching or whitening the skin and skin appendages of a healthy individual.

Description

Cosmetic use of compounds derived from oleanolic acid

[0001] The invention relates to the use of compounds derived from oleanolic acid in cosmetics, and in particular in the context of skin colour modulation.

[0002] Controlling skin pigmentation is playing an increasingly important role in cosmetic research. Indeed, for socio-cultural or aesthetic reasons, there is a growing enthusiasm for cosmetic products designed to lighten skin color or to remove or reduce spots caused by pigmentation disorders.

[0003] The pigmentation of the skin and hair of mammals, and particularly humans, is due to the presence of melanins, complex polymer pigments whose color can vary from brown-black (eumelanin) to red (pheomelanin). The biosynthesis of melanin, melanogenesis, is carried out by specialized cells called melanocytes, which are found in large numbers in the eyes, hair follicles, and skin. The different stages of melanogenesis are now relatively well understood.

[0004] In the various stages of melanogenesis, only the first two reactions are controlled by a metalloenzyme called tyrosinase, which oxidizes tyrosine to dihydroxyphenylalanine (DOPA), and then to dopaquinone. Therefore, if one wishes to inhibit or stimulate melanogenesis, the easiest method of regulation is by modifying the activity of tyrosinase.

[0005] Several depigmenting agents are known to inhibit melanin biosynthesis by acting on tyrosinase. Some, such as kojic acid, suppress tyrosinase synthesis or maturation by chelating the copper ions essential for its function. Others directly inhibit tyrosinase activity by acting as substrate analogs, occupying the enzyme's active site and competing with tyrosine or DOPA. This is probably the case for hydroquinone, a well-known depigmenting agent whose use is now strictly regulated due to its high cytotoxicity, as well as for arbutin (4-hydroxyphenyl-β-D-glucopyranoside), a depigmenting compound extracted from several plants (see French patent 94 08750).

[0006] Japanese patent application no. 63-8317 discloses an external agent with skin-lightening properties containing chelidonic acid as its active ingredient. This acid, found in various plants and particularly extractable from celandine, has long been prepared by organic synthesis from acetone and ethyl oxalate.

[0007] The object of the present invention is therefore a depigmenting cosmetic or dermatological composition containing, as an active principle, a compound derived from oleanolic acid and its use for lightening the skin or hair, or for treating pigment spots.

[0008] Therefore, the invention relates to the cosmetic and non-therapeutic use of a composition comprising, consisting essentially of, or consisting of a compound of the following formula I: (I)

[0009] or its enantiomers, isomers or diastereomers, or a mixture thereof,

[0010] or a salt or a solvate thereof,

[0011] where R1 and R2, independently of each other, are a C3 polyol, a C4-C6 sugar, a uronic acid, a glycoside, in particular glucoside, or R1 is OH and R2 is a C4-C6 sugar, or a C3 polyol, or R1 is a C4-C6 sugar, or a C3 polyol, and R2 is OH,

[0012] provided that if the composition is an extract of Aralia taibaiensis, said composition comprises more than 2.5% by mass relative to the total mass of said composition of said compound,

[0013] for the discoloration or bleaching of the skin or hair of a healthy individual.

[0014] The invention is based on the surprising observation that compounds of formula I act on the depigmentation of the skin or skin appendages by inhibiting melanin production. Skin appendages are integumentary structures derived from the ectoderm and characterized by a high degree of keratinization. In humans, and within the scope of this invention, the principal skin appendages are hair, body hair, and nails.

[0015] The compounds used in the context of the invention have one or more asymmetric carbons and may be in the form of one of their enantiomers, and / or one of their isomers, or a mixture of these, in particular in the form of a racemic mixture.

[0016] Uronic acids are chemical compounds obtained by oxidation of the last carbon of simple sugars. They are components of glycosaminoglycans, essential constituents of extracellular matrices.

[0017] A glucoside is a glycoside derived from glucose. The simplest glucosides are alkyl ethers, obtained by reacting hydrochloric acid with alcoholic glucose solutions. A more advanced synthetic method involves dissolving anhydrous glucose in methanol containing hydrochloric acid, producing a mixture of α- and β-methylglucosides.

[0018] In the invention, "C3-C6 sugars" refers to sugars, aldoses or ketoses, having 4, 5 or 6 carbon atoms. These sugars, or monosaccharides, are in their linear or cyclic form, according to the furanose or pyranose form, and in the a or b isomeric form, according to the terminology well known to those skilled in the art.

[0019] In the invention, "C3 polyol" refers in particular to a diol or polyol comprising 3 carbon atoms such as glycerol, dihydroxyacetone, or glyceraldehyde.

[0020] It is also envisaged in the context of the invention of heteroside derivatives, known as glycosides, of C3-C6 sugars.

[0021] In the invention, "discoloration or whitening of the skin or skin appendages of a healthy individual" refers to a change in skin color from a darker to a lighter complexion, through a reduction in skin phototype, at least in one area. The same applies to body hair.

[0022] Regarding skin, there are six skin phototypes that can be defined as follows: phototype 1, which corresponds to very fair skin, associated with freckles, often in red-haired or blond individuals; phototype 2, which also corresponds to very fair skin with freckles appearing after sun exposure, often in brown or blond individuals; phototype 3, which corresponds to fair skin often in brown or blond individuals; phototype 4, which corresponds to matte skin, in brown or blond individuals; phototype 5, which corresponds to dark skin; and phototype 6, which corresponds to black skin.

[0023] Within the framework of the invention, bleaching using the aforementioned compounds is desirable to move from a phototype n to a phototype at least n-1, or even n-2 or more.

[0024] Also, we will talk about discoloration in the context of a transition from a phototype 4 to a phototype 3 or a phototype 2, or even a phototype 1.

[0025] For hair, depending on the melanin concentration, there are eight natural colors: black (including raven black), the most common color worldwide; brown, the second most common; auburn; chestnut; red; strawberry blonde; blonde; and completely depigmented white. As with skin, in the context of this invention, bleaching with the aforementioned compounds is intended to move from a dark color to a lighter one. For example, it is possible to go from black to the next shade below it, i.e., brown, or even lighter.

[0026] Advantageously, the compounds according to the invention, for the aforementioned use, are isolated from their natural context and purified. Furthermore, it is advantageous that when the composition according to the invention comprises a plant extract containing a compound according to the invention, the proportions of said compound are not those obtained by extracting said compound from the plant.

[0027] According to the present invention, the compounds represented by formula I or their salts may be in anhydrous form or as a solvate, such as a hydrate or other. The term "solvate" here refers to a phenomenon whereby solute molecules or ions in a solution strongly attract neighboring solvent molecules to form a large group of molecules. When the solvent is water, it is called a "hydrate." The solvent may be a hydrate or a non-hydrate. Preferably, the solvates are pharmaceutically acceptable. For the non-hydrate, alcohols (e.g., methanol, ethanol, n-propanol), dimethylformamide, and other similar substances may be used.

[0028] The compounds of the present invention and their salts can exist in several tautomeric forms, for example, as enol and imine, keto and enamine, and mixtures of these forms. In solution, a tautomer is present as a mixture of tautomeric complexes. In the case of a solid form, one type of tautomer is generally present in a dominant proportion. In this respect, even if only one type of tautomer is described, the present invention encompasses all the types of tautomers of the compounds of the present invention.

[0029] The present invention includes all types of stereoisomers of the compounds of the present invention that are represented by formula (I) (for example, enantiomer, diastereomer (including the cis and trans geometric isomer)), racemate of the isomer, and mixtures thereof. For example, compounds having formula (I) of the present invention may have one or more asymmetric centers, and the present invention includes a racemic mixture, a mixture of diastereomers, and an enantiomer of this compound.

[0030] When the compounds of the present invention are obtained in free form, they can be converted into a salt, a hydrate or a solvate which can be formed from the compounds according to a method generally known in the art.

[0031] Furthermore, when the compounds of the present invention are obtained in the form of a salt, a hydrate or a solvent of the compounds, they can be converted to the free state by a method generally known in the art.

[0032] The present invention includes all isotopes of the compounds represented by formula I. Isotopes of the compounds of the present invention refer to compounds of the present invention in which at least one atom is replaced by an atom having the same atomic number (i.e., the number of protons) but a different mass number (the sum of the number of protons and the number of neutrons). Isotopes included in the compounds of the present invention include, for example, a hydrogen atom, a carbon atom, an oxygen atom, such as 2 H, 3 H, 13 C, 14 C, 17 O and 18 O. In particular, a radioisotope that decays by emitting radiation, for example3 H and 14 C, is useful for determining the distribution of a pharmaceutical agent or compound in living tissue, etc. In contrast, a stable isotope does not degrade and remains practically unchanged without exhibiting radioactivity, and can therefore be used safely. The isotopes of the compounds of the present invention can be converted by replacing a chemical reagent used for synthesis with a chemical reagent comprising a corresponding radioisotope according to a method generally known in art.

[0033] The composition according to the invention may essentially consist of the aforementioned compound, meaning that the active substance having a bleaching effect is the aforementioned compound. This composition does not include any other compound having a depigmenting effect.

[0034] The composition may contain one or more other compounds which do not participate in the discoloration of the skin or hair but contribute to the cosmetic properties of stability, applicability, dispersibility... necessary for the use of said composition.

[0035] It may therefore be advantageous to introduce into the composition used according to the invention at least one compound chosen from: desquamating agents; soothing agents, organic or inorganic photoprotective agents, moisturizing agents; anti-glycation agents; NO-synthase inhibitors; agents stimulating the synthesis of dermal or epidermal macromolecules and / or preventing their degradation; agents stimulating the proliferation of fibroblasts and / or keratinocytes or stimulating the differentiation of keratinocytes; muscle relaxants and / or dermo-relaxing agents; tightening agents; anti-pollution and / or anti-radical agents; agents acting on microcirculation; agents acting on the energy metabolism of cells; and mixtures thereof.Examples of such additional compounds include: retinol and its derivatives such as retinyl palmitate; ascorbic acid and its derivatives such as magnesium ascorbyl phosphate and ascorbyl glucoside; tocopherol and its derivatives such as tocopheryl acetate; nicotinic acid and its precursors such as nicotinamide; ubiquinone; glutathione and its precursors such as L-2-oxothiazolidine-4-carboxylic acid; plant extracts, including plant proteins and their hydrolysates, as well as phytohormones; marine extracts such as seaweed extracts; bacterial extracts; sapogenins such as diosgenin and extracts of wild yam containing it; ceramides; hydroxy acids such as salicylic acid and n-octanoyl-5-salicylic acid; resveratrol; oligopeptides and pseudodipeptides and their acylated derivatives; manganese and magnesium salts, in particular gluconates; and mixtures thereof.By desquamating agent, we mean any compound capable of acting: - either directly on desquamation by promoting exfoliation, such as β-hydroxy acids, in particular salicylic acid and its derivatives (including n-octanoyl 5-salicylic acid); alpha-hydroxy acids (or AHAs), such as glycolic, citric, lactic, tartaric, malic or mandelic acids; urea; gentisic acid; oligofucoses; cinnamic acid; Saphora japonica extract; resveratrol; - or on the enzymes involved in desquamation or the degradation of corneodesmosomes, glycosidases, stratum corneum chymotryptic enzym (SCCE) or even other proteases (trypsin, chymotrypsin-like).Examples of chelating agents for mineral salts include: EDTA; N-acyl-N,N',N' ethylenediaminetriacetic acid; aminosulfonic compounds, particularly (N-2-hydroxyethylpiperazine-N-2-ethane) sulfonic acid (HEPES); derivatives of 2-oxothiazolidine-4-carboxylic acid (procysteine); glycine-type alpha amino acid derivatives (as described in EP-0 852 949, as well as sodium methylglycine diacetate marketed by BASF under the trade name TRILON M); honey; and sugar derivatives such as O-octanoyl-6-D-maltose and N-acetyl glucosamine.

[0036] The desquamating agents are generally present in the composition according to the invention in proportions ranging from 0.01 to 15% by weight, preferably ranging from 0.1 to 10% by weight, relative to the total weight of the composition. Examples of soothing agents that can be used in the composition according to the invention include: pentacyclic triterpenes and plant extracts (e.g., Glycyrrhiza glabra) containing them, such as β-glycyrrhetinic acid and its salts and / or derivatives (glycyrrhetinic acid monoglucuronide, stearyl glycyrrhetinate, 3-stearoyloxy glycyrrhetic acid), ursolic acid and its salts, oleanolic acid and its salts, betulinic acid and its salts, an extract of Paeonia suffruticosa and / or lactiflora, salts of salicylic acid and in particular zinc salicylate, phycosaccharides from the company Codif, an extract of Laminaria saccharina, canola oil, bisabolol and chamomile extracts, allantoin,Seppic's Sepivital EPC (vitamin E and C diesterphosphoric acid), omega-3 unsaturated oils such as rosehip, blackcurrant, ecchium, and fish oils, plankton extracts, capryloyl glycine, Seppicalm VG (sodium palmitoylproline and Nymphaea alba) from Seppic, Pygeum extract, Boswellia serrata extract, Centipeda cunningham extract, Helianthus annuus extract, Linum usitatissimum extract, tocotrienols, Cola nitida extract, piperonal, clove extract, Epilobium angustifolium extract, aloe vera, Bacopa moniera extract, phytosterols, cortisone, hydrocortisone, indomethacin, and beta methasone.

[0037] The soothing agents are generally present in the composition used according to the invention in proportions ranging from 0.01 to 15% by weight, preferably ranging from 0.1 to 10% by weight, relative to the total weight of the composition.

[0038] Organic photoprotective agents are notably selected from among anthranilates; cinnamic derivatives; dibenzoylmethane derivatives; salicylic derivatives; camphor derivatives; triazine derivatives such as those described in patent applications US4367390, EP863145, EP517104, EP570838, EP796851, EP775698, EP878469, EP933376, EP507691, EP507692, EP790243, EP944624; benzophenone derivatives; β,β-diphenyl acrylate derivatives; benzotriazole derivatives; benzalmalonate derivatives; benzimidazole derivatives; imidazolines; bis-benzoazolyl derivatives as described in patents EP669323 and US2,463,264; p-aminobenzoic acid (PABA) derivatives; methylene bis-(hydroxyphenyl benzotriazole) derivatives as described in applications US5237071, US5166355, GB2303549, DE19726184 and EP893119; and filter polymers and filter silicones such as those described in particular in application WO-93 / 04665;alpha-alkylstyrene derived dimers such as those described in patent application DE19855649.;

[0039] Inorganic photoprotective agents can be selected from pigments or nanopigments (average primary particle size generally between 5 nm and 100 nm, preferably between 10 nm and 50 nm) of metal oxides, coated or uncoated, such as titanium dioxide nanopigments (amorphous or crystallized as rutile and / or anatase), iron, zinc, zirconium, or cerium, all of which are well-known UV photoprotective agents. Conventional coating agents include alumina and / or aluminum stearate. Such metal oxide nanopigments, coated or uncoated, are described in particular in patent applications EP518772 and EP518773.

[0040] Photoprotective agents are generally present in the composition used according to the invention in proportions ranging from 0.1 to 20% by weight, preferably ranging from 0.2 to 15% by weight, relative to the total weight of the composition.

[0041] Advantageously, the invention relates to the aforementioned use, where said sugar is selected from erythrose, threose, erythrulose, ribose and deoxyribose, arabinose, xylose, lyxose, ribulose, xylulose, allose, altrose, galactose, glucose, gulose, idose, mannose, talose, fructose, psicose, sorbose and tagadose, and deoxyhexoses, namely fucose and rhamnose.

[0042] The aforementioned C4-C6 sugars can be in their L or D form, and in linear or cyclic form, including furanoside or pyranoside; in alpha or beta form, where possible.

[0043] Advantageously the invention relates to the aforementioned use, where the uronic acids are beta-D-Glucuronic acid, alpha-D-Glucoronic acid, alpha-D-glucopyranuronic acid, beta-D-glucopyranuronic acid, alpha-D-galactopyranuronic acid, beta-D-galactopyranuronic acid, beta-L-Glucuronic acid, alpha-L-Glucuronic acid, alpha-L-glucopyranuronic acid, beta-L-glucopyranuronic acid, alpha-L-galactopyranuronic acid, beta-L-galactopyranuronic acid, or hexopyranuronic acid.

[0044] Glycosides (or heterosides) are molecules formed by the condensation of a sugar (a monosaccharide, then called a glycone) and a non-carbohydrate substance (called an aglycone or genin). These two components are joined by a glycosidic bond, the type of which defines the glycoside's classification. The bond can be O-type (defining an O-glycoside), N-type (defining a glycosylamine), S-type (defining a thioglycoside, e.g., glucosinolate), or C-type (defining a C-glycoside). This bond can be broken by hydrolysis, which separates the glycone and the genin.

[0045] It is advantageous that these glycosides are derivatives of glucose, but also of galactose, mannose, lactose or fructose.

[0046] It is in the invention that the aforementioned residues R1 and R2 are interchangeable with each other.

[0047] It is also conceivable that, within the context of the use described above, the composition may comprise the compound as defined above, in association with one or more compounds known in the art, such as 4-Butylresorcinol, 4-Isopropylcatechol, 4-Methoxyphenol, Acetyl Glucosamine, Acetyl Glycyl Beta-Alanine, Acetyl Hexapeptide-1, Acetyl Rheum Rhaponticum Root Extract, Acetyl Zingerone, Ajuga Reptans Cell Culture Extract, Alloferon-1, Alpha-Arbutin, Alpha-Hydroxy Acids (AHAs), Alpha Tocopherol, Alpha Tocopherol Ferulate, Ammonium Persulfate, Aminopropyl Ascorbyl Phosphate, Angelica Dahurica Root Extract, Arbutin, Arctostaphylos Uva Ursi Leaf Powder, Arctostaphylos Uva-Ursi Leaf Extract, Artocarpus Altilis Wood Extract, Artocarpus Heterophyllus Seed Extract, Artocarpus Lakoocha Wood Extract, Ascorbic Acid, Ascorbyl Glucoside, Ascorbyl Linoleate, Azelaic Acid, Azeloyl Diglycinate, Bakuchiol, Bellis Perennis Extract,le Betaphycus Gelatinum Extract, le Betula Papyrifera Bark Extract, le Bis-Ethylhexyl Hydroxydimethoxy Benzylmalonate, le Buddleja Axillaris Leaf Extract, le Calcium Sulfate Hydrate, le Camellia Sinensis Catechins (EGCG, le ECG), le Carica Papaya Extract, le Centaureidin, le Ceramide, le Cetyl Tranexamate Mesylate, le Citrus Unshiu Peel Extract, le Cysteamine, le Cystoseira Tamariscifolia Extract, le Curcuma Longa Root Extract, le Copper Tripeptide-1, le Cyclohexanediol Bis-Ethylhexanoate, le Decapeptide-12, le Decapeptide-15, le Deoxyarbutin, le Dipotassium Glycyrrhizate, le Difluorocyclohexyloxyphenol, le Diglucosyl Gallic Acid, le Dimethylmethoxy Chromanyl Palmitate, le Disodium EDTA, le Dioxide de Titane (Titanium Dioxide), le Ellagic Acid, le Ethyl Ascorbic Acid, le Gallic Acid, le Gentisic Acid, le Gigawhite, le Glabridin, le Glutathione (Reduced, le Ethyl Ester, le Liposomal, le Acetylated Forms), le Glycine Soja Seed Extract, le Glyceryl Glucoside, le Glycolic Acid,le Ginkgo Biloba Extract, le Ginseng Extract, le Green Tea Extract, l’Hesperidin, l’Hexapeptide-2, l’Hexylresorcinol, l’Hibbertia Scandens Leaf Extract, l’Hibiscus Rosa-Sinensis Leaf Extract, l’Honokiol, l’Human Adipose Derived Stem Cell Conditioned Media, l’Hydrolyzed Codonopsis Pilosula Root Extract, l’Hydrolyzed Xanthomonas Campestris Polysaccharide, l’Hydroquinone, l’Hydroxystilbene (Resveratrol, l’Oxyresveratrol, l’Pterostilbene), l’Hydroxyapatite, l’Hydroxycinnamic Acid & Derivatives, l’Hydroxyphenoxy Propionic Acid, l’Hydroxyphenyl Propionamide, l’Hydroxyresveratrol, l’Hydroxytyrosol, l’Isobutylamido Thiazolyl Resorcinol, le Kalanchoe Spathulata Extract, le Kojic Acid, le Lactobacillus / Nelumbo Nucifera Seed Ferment Filtrate, le Lactobacillus / Rice Bran / Saccharomyces / Camellia Sinensis Leaf Extract Ferment, le Lentinus Edodes Mycelium Extract, le Linoleic Acid, le Lysine Azelate, le Magnesium Ascorbyl Phosphate, le Magnolol, le Menaquinone-7, le Mequinol (4-Hydroxyanisole),le Melanostatine-5 (Nonapeptide-1), le Melatonin, le Methimazole, le Methoxysalicylates (Potassium Methoxysalicylate), le Methoxy PEG-45 Thioctate, le Methylophiopogonanone B, le Monobenzyl Ether of Hydroquinone, le Morus Alba Root Extract, le Morus Nigra Extract, le Mulberry Extract, le Myristyl Theobroma Grandiflorum Seedate, le N-Acetyl Glucosamine (NAG), le N-Acetyl-4-S-Cysteminylphenol, le Nelumbo Nucifera Extract, le Niacinamide, le Nonapeptide-12, le Oleanolic Acid Derivatives, le Oligopeptide-34, le Ozonized Ethyl Oleate, le Palmitoyl Grapevine Shoot Extract, le Paeoniflorin, le Pentapeptide-15, le Phenylalanine & Derivatives, le Phenylethyl Resorcinol, le Phytic Acid, le Prunus Armeniaca Kernel Extract, le Punicic Acid, le Rhodopseudomonas, le Salicylic Acid, le Sanguisorba Officinalis Root Extract, le Sh-Pentapeptide-19, le Sodium Palmitoyl Proline, le Sophora Flavescens Root Extract, le Sulforaphane, le Tannic Acid, le Terminalia Ferdinandiana Fruit Extract,Tetrahexyldecyl Ascorbate, Tetrahydropiperine, Tranexamic Acid, Tranexamoyl Tetrapeptide-2, Tranexamoyl Dipeptide-22, Tranexamoyl Dipeptide-23, Tranexamoyl Tripeptide-1, Trimethoxybenzyl Acetylsinapate, Triticum Vulgare Germ, Ulva Pertusa Extract, Undecylenoyl Phenylalanine, Withania Somnifera Leaf Extract, Glycyrrhiza Glabra (Licorice) Root Extract, Retinoids (Retinol, Retinaldehyde, Tretinoin, Adapalene), Corticosteroids (Hydrocortisone, Dexamethasone, etc.), Aloesin, Atractylodes Japonica Root Oil, Thiopyridinone Derivatives (2-Mercaptonicotinoyl Glycine & salts).

[0048] Advantageously, the invention relates to the aforementioned use, where said compound has the following formula Ia: (Ia)

[0049] where R1 and R2 are as defined above.

[0050] Even more advantageously the invention relates to the aforementioned use, where the compound is chosen from the compounds described in Table 1.

[0051] The most advantageous compounds according to the invention, used in the context of skin depigmentation or whitening, are compounds with the following structure: (I)

[0052] where R1 is such that the rows in table 1 are defined by and R2 is such that the columns in table 1 are defined by.

[0053] R2 →R1↓Erythrosele threosele erythrulosele ribosele deoxyriboseArabinosele xylosele lyxosele ribulosele xyluloseAlloseAltrosele galactosele glucosele guloseIdosele mannosele talosele fructosele psicosele sorbosele tagatosele fucosele rhamnoseGlucuronic acidGlucopyranuronic acidGalcopyranuronic acidHexopyranuronic acidGlycerolErythrose+++++++++++++++++++++++++++++threose+++++++++++++++++++++++++++++Erythrulose+++++++++++++++++++++++++++++++ribose+++++++++++++++++++++++++++++++ deoxyribose+++++++++++++++++++++++++++++arabinose+++++++++++++++++++++++++++++xylose+++++++++++++++++++++++++++++++lyxose+++++++++++++++++++++++++++++++ribulose+++++++++++++++++++++++++++++++++xylulose+++++++++++++++++++++++++++++++allose+++++++++++++++++++++++++++++Altrose+++++++++++++++++++++++++++++Galcose+++++++++++++++++++++++++++++glucose+++++++++++++++++++++++++++++++gulose+++++++++++++++++++++++++++++++idose+++++++++++++++++++++++++++++++mannose++++++++++++++++++++++++++++ talose++++++++++++++++++++++++++++++ fructose++++++++++++++++++++++++++++++++ psicose++++++++++++++++++++++++++++++++ sorbose++++++++++++++++++++++++++++++ tagadose++++++++++++++++++++++++++++++++ fucose++++++++++++++++++++++++++++++ rhamnose++++++++++++++++++++++++++++++ Glucuronic acid++++++++++++++++++++++++++++++ Glucopyranuronic acid++++++++++++++++++++++++++++++ galactopyranuronic acid++++++++++++++++++++++++++++++ Hexopyranuronic acid++++++++++++++++++++++++++++Glycerol++++++++++++++++++++++++++++Dihydroxyacetone++++++++++++++++++++++++++++++Glyceraldehyde+++++++++++++++++++++++++++++++

[0054] This means, for example, that if R1 is erythrose, R2 can be erythrose; threose; erythrulose; ribose; deoxyribose; arabinose; xylose; lyxose; ribulose; xylulose; allose; altrose; galactose; glucose; gulose; idose; mannose; talose; fructose; psicose; sorbose; tagatose; fucose; rhamnose; glucuronic acid; glucopyranuronic acid; galactopyranuronic acid or hexopyranuronic acid.

[0055] Therefore, there are 899 advantageous compounds derived from formula I.

[0056] The case where R1 is a sugar as described above and R2 is a polysaccharide or disaccharide, or vice versa, is also described, in particular

[0057] R1 is erythrose; threose; erythrulose; ribose; deoxyribose; arabinose; xylose; lyxose; ribulose; xylulose; allose; altrose; galactose; glucose; gulose; idose; mannose; talose; fructose; psicose; sorbose; tagatose; fucose; rhamnose; glucuronic acid; glucopyranuronic acid; galactopyranuronic acid or hexopyranuronic acid or glycerol and R2 is sucrose,

[0058] R2 is erythrose; threose; erythrulose; ribose; deoxyribose; arabinose; xylose; lyxose; ribulose; xylulose; allose; altrose; galactose; glucose; gulose; idose; mannose; talose; fructose; psicose; sorbose; tagatose; fucose; rhamnose; glucuronic acid; glucopyranuronic acid; galactopyranuronic acid or hexopyranuronic acid or glycerol and R1 is sucrose,

[0059] R1 is erythrose; threose; erythrulose; ribose; deoxyribose; arabinose; xylose; lyxose; ribulose; xylulose; allose; altrose; galactose; glucose; gulose; idose; mannose; talose; fructose; psicose; sorbose; tagatose; fucose; rhamnose; glucuronic acid; glucopyranuronic acid; galactopyranuronic acid or hexopyranuronic acid or glycerol and R2 is lactose,

[0060] R2 is erythrose; threose; erythrulose; ribose; deoxyribose; arabinose; xylose; lyxose; ribulose; xylulose; allose; altrose; galactose; glucose; gulose; idose; mannose; talose; fructose; psicose; sorbose; tagatose; fucose; rhamnose; glucuronic acid; glucopyranuronic acid; galactopyranuronic acid or hexopyranuronic acid or glycerol and R1 is lactose,

[0061] R1 is erythrose; threose; erythrulose; ribose; deoxyribose; arabinose; xylose; lyxose; ribulose; xylulose; allose; altrose; galactose; glucose; gulose; idose; mannose; talose; fructose; psicose; sorbose; tagatose; fucose; rhamnose; glucuronic acid; glucopyranuronic acid; galactopyranuronic acid or hexopyranuronic acid or glycerol and R2 is maltose,

[0062] R2 is erythrose; threose; erythrulose; ribose; deoxyribose; arabinose; xylose; lyxose; ribulose; xylulose; allose; altrose; galactose; glucose; gulose; idose; mannose; talose; fructose; psicose; sorbose; tagatose; fucose; rhamnose; glucuronic acid; glucopyranuronic acid; galactopyranuronic acid or hexopyranuronic acid or glycerol and R1 is maltose,

[0063] R1 is sucrose and R2 is sucrose.

[0064] R1 is sucrose and R2 is lactose.

[0065] R1 is sucrose and R2 is maltose.

[0066] R1 is lactose and R2 is lactose.

[0067] R1 is lactose and R2 is maltose.

[0068] R1 is maltose and R2 is maltose.

[0069] Other compounds such as R1 (dihydroxyacetone) and R2 (glyceraldehyde), not listed in the table above, are also covered. Similarly, compounds such as

[0070] R2 is dihydroxyacetone, and R1 is any one of the following: erythrose, threose, erythrulose, ribose, deoxyribose, arabinose, xylose, lyxose, ribulose, xylulose, allose, altrose, galactose, glucose, gulose, idose, mannose, talose, fructose, psicose, sorbose, tagadose, fucose, rhamnose, glucuronic acid, glucopyranuronic acid, galactopyranuronic acid, hexopyranuronic acid, and glycerol; or R2 is glyceraldehyde and R1 is any one of the following: erythrose, threose, erythrulose, ribose, deoxyribose, arabinose, xylose, lyxose, ribulose, xylulose, allose, altrose, galactose, glucose, gulose, idose, mannose, talose, fructose, psicose, sorbose, tagadose, fucose, rhamnose, glucuronic acid, glucopyranuronic acid, galactopyranuronic acid, hexopyranuronic acid and glycerol are also covered by the invention.The alpha and beta isomers of these glucuronic acids are also covered in the present invention.

[0071] Even more advantageously, the compounds according to the invention, used in the context of skin depigmentation or whitening, are compounds with the following structure: (Ia)

[0072] where R1 is such that the rows in table 1 are defined by and R2 is such that the columns in table 1 are defined by.

[0073] The following are some beneficial compounds: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , And .

[0074] Even more advantageously, the invention relates to the aforementioned use, said compound being Calenduloside F of the following formula Ib: (Ib).

[0075] Calenduloside F is known by its IUPAC name as (2S,3S,4S,5R,6R)-6-[[(3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-4,4,6a,6b,11,11,14b-heptamethyl-8a-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxycarbonyl-1,2,3,4a,5,6,7,8,9,10,12,12a,14,14a-tetradecahydropicen-3-yl]oxy]-3,4,5-trihydroxyoxane-2-carboxylic acid and the CAS number: 51415-02-2.

[0076] It can be obtained by purification from Calendula officinalis flowers, and is offered for sale by several manufacturers.

[0077] The invention also relates to a composition for topical application, or in a form suitable for topical application, comprising from 0.00001 to 1% by mass relative to the total mass of the composition of one of the aforementioned compounds, in particular Calenduloside F, of formula Ib.

[0078] The composition comprising a compound as defined above, may be associated with a cosmetically acceptable vehicle, in particular dermatologically acceptable.

[0079] In the invention, the terms "cosmetically acceptable" mean that the composition or components thereof are suitable for use in contact with human skin and / or mucous membranes without undue toxicity, incompatibility, instability, allergic response, or their equivalents.

[0080] The cosmetically acceptable vehicle may be chosen from water, allantoin, glycerin, methylpropanediol; this list is not exhaustive.

[0081] The composition is in a form chosen from the group comprising an oil-in-water or water-in-oil emulsion, or a mixture of these emulsions. The cosmetic composition may, for example, be in a form chosen from the group comprising an aqueous or hydroalcoholic gel, an aqueous or hydroalcoholic cream, and an aqueous or hydroalcoholic lotion. These usable formulations are known to formulators in the prior art.

[0082] The composition may be in a form chosen from an ointment, cream, oil, milk, pomade, powder, soaked pad, solution, gel, serum, balm, butter, lotion, suspension, soap or emulsion.

[0083] All these forms are cosmetically acceptable and allow for topical application.

[0084] The composition is such that the compound is in a proportion of 0.00001 to 10% by mass relative to the total mass of the composition, preferably 0.00001 to 5% by mass relative to the total mass of the composition, preferably 0.00001 to 3% by mass relative to the total mass of the composition, preferably 0.00001 to 2% by mass relative to the total mass of the composition and even more preferably 0.00001 to 1% by mass relative to the total mass of the composition.

[0085] Advantageously, the composition is applied to the skin, particularly in the form of a cream or lotion, in an amount between 0.0001 and 100 mg / cm² 2 / day of compound as defined previously.

[0086] The aforementioned composition is such that the compound and the vehicle can be used alone or in combination with other biologically active ingredients and / or substances that, in particular, allow the compound according to the invention to be accompanied, to constitute a specific formulation, or to preserve the compound over time; this list is not exhaustive. In other words, it can be any basic product found in conventional cosmetic compositions.

[0087] The composition according to the invention may further contain inert additives or combinations thereof, such as: wetting agents; preservatives; stabilizing agents; moisture-regulating agents; pH-regulating agents; osmotic pressure-modifying agents; emulsifying agents; and antioxidants. This list is not exhaustive.

[0088] Of course, a person skilled in the art will take care to choose the possible compound(s) to be added to these compositions and their respective quantities in such a way that the advantageous properties intrinsically attached to the present invention are not or substantially not altered by the addition, or additions, envisaged.

[0089] It is advantageous for the concentration of the compound to be in the range of 0.01µM to 15 mM, preferably 0.1µM to 1 mM, and especially 0.25µM to 500µM when the formulation is liquid, and not in solid form.

[0090] The invention also relates to a non-therapeutic cosmetic method of skin depigmentation or whitening comprising a step of topical application of a compound as defined above, or of the aforementioned composition on the skin of a healthy individual.

[0091] To depigment the skin of a healthy individual, that is, an individual not suffering from a dermatological condition affecting the skin from a pigmentary point of view, the method of the invention involves applying topically, to the area(s) to be depigmented, an effective amount of the aforementioned compound, or composition. This application may be daily, or more frequently.

[0092] The compound is advantageously in a proportion of 0.00001 to 10% by mass relative to the total mass of the composition, preferably from 0.00001 to 5% by mass relative to the total mass of the composition, preferably from 0.00001 to 3% by mass relative to the total mass of the composition, preferably from 0.00001 to 2% by mass relative to the total mass of the composition and even more preferably from 0.00001 to 1% by mass relative to the total mass of the composition.

[0093] It is advantageous for the application to be carried out with a compound concentration of the order of 0.01µM to 10mM, preferably of 0.1µM to 1mM, and in particular of 0.25 to 500 µM, in a liquid and not solid application.

[0094] It is also recommended not to expose oneself to the sun during this application in order to limit the effects of ultraviolet rays which could diminish the effects of the compounds of the invention or the composition according to the invention.

[0095] The invention relates in addition to a cosmetic composition intended for use in a process for the prevention or treatment of a melanin hyperpigmentation disease, the cosmetic composition comprising a compound as defined above.

[0096] It has been observed that the aforementioned compounds can prevent and / or treat hyperpigmentation spots on healthy skin or skin appendages, i.e., freckles, melasma / chloasma, dark spots or brown spots, lentigo, Addison's syndrome, vitiligo, post-inflammatory hyperpigmentation due to abrasion, burn, scar, dermatosis, contact allergy; nevi, genetically determined hyperpigmentation, hyperpigmentation of metabolic or drug origin, melanoma, or any other hyperpigmented lesions. This list is not exhaustive.

[0097] Lentigo is a specific form of hyperpigmentation, strongly associated with skin aging and sun exposure. Lentigo is also sometimes called solar lentigo, actinic lentigo, senile lentigo, or simply "age spots."

[0098] Chloasma is a specific form of hyperpigmentation, also called melasma or the "mask of pregnancy" when it specifically affects pregnant women. Many women experience chloasma even when not pregnant, due to taking hormonal contraceptives. This is sometimes referred to as "pill-related melasma."

[0099] The compositions according to the invention may further include any additive commonly used in the cosmetic or pharmaceutical field, such as sequestrants, antioxidants, preservatives, fillers, electrolytes, humectants, colorants, common inorganic or organic bases or acids, perfumes, essential oils, cosmetic actives, moisturizers, vitamins, essential fatty acids, sphingolipids, and skin-soothing and protective agents, such as allantoin. Naturally, those skilled in the art will take care to choose these optional additional compounds, and / or their quantity, in such a way that the advantageous properties of the composition according to the invention are not, or not substantially, altered.

[0100] The aforementioned compound is also envisaged within the framework of the invention for its use in the treatment or prevention of a skin hyperpigmentation disease.

[0101] The pharmaceutical form of the aforementioned compound for its aforementioned use is in particular as described above.

[0102] Furthermore, a method for the therapeutic treatment or prevention of a skin hyperpigmentation condition is being considered, comprising a step of topical application to the skin of an individual suffering from hyperpigmentation, of an effective quantity of a compound as defined above. The effective quantity is as defined above for cosmetic purposes.

[0103] The invention also relates to a compound of formula (I),

[0104] Where R1 and R2 independently of each other are glycerol, glucose, or glucuronic acid; or R1 is OH and R2 is a C3 polyol or a C4-C6 sugar, provided that the C6 sugar is not glucose; or R1 is a C3 polyol or a C4-C6 sugar, provided that the C6 sugar is not glucose, and R2 is OH, and

[0105] provided that the compound is not the compound with the formula: .

[0106] Within the scope of the invention, the aforementioned compounds are novel. It is understood that Calenduloside F is not covered with respect to the compounds as such.

[0107] Advantageously, the invention relates to the aforementioned compound, said compound being selected from the following compounds: , , , , , , , , , , , , , , , , , , , And . Brief description of the figures

[0108] The invention will be better understood upon reading the following description, given solely by way of example and made with reference to the accompanying drawings in which:

[0109] This is a photograph of a control skin section stained with hematoxylin and eosin (H&E) on day 0 (D0). The bar represents 100 µm.

[0110] Figure 1 shows photographs of three sets of skin sections (1, 2, and 3) stained with hematoxylin and eosin (H&E) on day 7 (D7). A: untreated (control), B: treated with UA and UVB (UVA 2.25 J / cm²). 2 and UVB 25mJ / cm 2), C: treated with UV (conditions B and with compound 1 at a concentration of 2 µM, D: treated with UV (conditions B and with compound 1 at a concentration of 0.5 µM, E: treated with UV (conditions B and with compound 1 at a concentration of 0.125 µM.

[0111] Figure 1 shows photographs of three sets of skin sections (1, 2, and 3) labeled with an anti-tyrosinase antibody on day 7 (D7) and detected by fluorescence. In the control, cell nuclei are labeled with DAPI. Arrows indicate cells positively labeled with anti-tyrosinase. A: untreated (control), B: treated with UA and UVB (UVA 2.25 J / cm²). 2 and UVB 25mJ / cm 2 ).

[0112] Figure 1 shows photographs of three sets of skin sections (1, 2, and 3) labeled with an anti-tyrosinase antibody on day 7 (D7) and detected by fluorescence. In the control, cell nuclei are labeled with DAPI. Arrows indicate cells positively labeled with anti-tyrosinase. A: untreated (control), B: treated with UA and UVB (UVA 2.25 J / cm²). 2 and UVB 25mJ / cm 2 ), C: treated with UV (conditions B and with compound 1 at a concentration of 2 µM, D: treated with UV (conditions B and with compound 1 at a concentration of 0.5 µM, E: treated with UV (conditions B and with compound 1 at a concentration of 0.125 µM.

[0113] Figure 1 represents a histogram quantifying the percentage of tyrosinase-positive cells as means ± standard deviation of the results shown in Figure 2 (*, p < 0.05; **, p < 0.01). A: untreated (control), B: treated with UA and UVB (UVA 2.25 J / cm²). 2 and UVB 25mJ / cm 2), C: treated with UV (conditions B and with compound 1 at a concentration of 2 µM, D: treated with UV (conditions B and with compound 1 at a concentration of 0.5 µM, E: treated with UV (conditions B and with compound 1 at a concentration of 0.125 µM.

[0114] Figure 1 shows photographs of three sets of skin sections (1, 2, and 3) demonstrating melanin expression by Fontana-Masson staining on day 7 (D7). A: untreated (control), B: treated with UA and UVB (UVA 2.25 J / cm²). 2 and UVB 25mJ / cm 2 ).

[0115] Figure 1 shows photographs of three series of skin sections (1, 2, and 3) demonstrating melanin expression by Fontana-Masson staining on day 7 (D7). A: treated with UA and UVB (UVA 2.25 J / cm²) 2 and UVB 25mJ / cm 2), B: treated with UV (conditions B and with compound 1 at a concentration of 2 µM, C: treated with UV (conditions B and with compound 1 at a concentration of 0.5 µM, D: treated with UV (conditions B and with compound 1 at a concentration of 0.125 µM.

[0116] Figure 1 represents a histogram quantifying melanin as positive spots in the total basal epidermis in the form of means + / - standard deviation of the results shown in Figure 2. A: untreated (control), B: treated with UA and UVB (UVA 2.25 J / cm2 and UVB 25 mJ / cm2), C: treated with UV (conditions B and with compound 1 at a concentration of 2 µM), D: treated with UV (conditions B and with compound 1 at a concentration of 0.5 µM), E: treated with UV (conditions B and with compound 1 at a concentration of 0.125 µM). Examples Example 1: Synthesis of compounds:

[0117] When compounds are not commercially available, they can be synthesized using methods known to those skilled in the art, including the following:

[0118] General information.

[0119] All reactions were carried out with anhydrous solvents in oven-dried glassware, under magnetic stirring and argon, unless otherwise specified. Chemicals were used as supplied, unless otherwise specified. Thin-layer chromatography (TLC) was performed on pre-coated silica gel plates (0.25–0.3 mm, Shanghai, China). The TLC plates were visualized by exposure to UV light or by staining with a sulfuric acid and ethanol solution. Silica gel column chromatography was performed on AR silica gel (100–200 mesh, Shanghai, China). Optical rotations (OR) were measured with a Rudolph Research Analytical Autopol I automatic polarimeter. NMR spectra were recorded with a Bruker Avance III 400 or Bruker Avance III 600 spectrometer.The 1H and 13C NMR spectra were calibrated against the residual proton and carbon signals of the solvents as internal references (CDCl3: δ3H = 7.26 ppm and δC = 77.2 ppm; CD3OD: δH = 3.31 ppm and δC = 49.0 ppm; pyridine-d5: δH = 7.20, 7.57, 8.72 ppm and δC = 123.4, 135.4, 149.8 ppm). Multiplicities are indicated as singlet (s), wide singlet (br s), doublet (d), triplet (t), quartet (q), doublet of doublets (dd), doublet of triplets (dt), triplet of doublets (td), or multiplet (m). All chemical shifts in NMR (δ) were recorded in ppm and coupling constants (J) were reported in Hz. High-resolution mass spectra were recorded on an ESI-TOF spectrometer.

[0120] 2.1 Synthesis of 28-O-2,3,4,6-tetra-O-benzoyl-β-D-glucopyranosyl oleanate 3-O-2,3-di-O-benzoyl-4,6-O-benzylidene-β-D-glucopyranoside11

[0121] A mixture of compound 9 (9.6 g, 15 mmol), compound 10 (10.35 g, 10 mmol), freshly activated 4 Å molecular sieve (25 g), N-iodo-succinimide (NIS, 4.5 g, 20 mmol), and TMSOTf (360 μL, 2 mmol) in anhydrous CH₂Cl₂ (200 mL) was stirred at 0°C for 30 min under argon. A solution of TMSOTf (360 μL, 2 mmol) was added. The mixture was stirred at the same temperature for 3 h, and then Et₃N was added to quench the reaction. The mixture was filtered through Celite, and the filtrate was washed with Na₂S₂O₃. The combined organic layers were dried over Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate: 15 / 1→5 / 1) to obtain 11(11.34 g, 76%) as a white solid.

[0122] [α] 25 D = +59.3 (c= 0.32, CHCl3 );

[0123] 1H RMN (400 MHz, CDCl3) δ 8,03-7,80 (m, 12 H), 7,56-7,25 (m, 23 H), 5.99-5.92 (m, 2 H), 5.78-5.70 (m, 3 H), 5.53-5.49 (m, 2 H), 5.26 (br s, 1 H), 4.83 (d, J = 8,0 Hz, 1 H), 4,54 (dd, J = 2,8, 12,0 Hz, 1 H), 4,46 (dd, J = 4,8, 12,4 Hz, 1 H), 4,39 (dd, J = 4,8, 10,4 Hz, 1 H), 4,26-4,22 (m, 1 H), 3,94-3,86 (m, 2 H), 3,69-3,63 (m, 1 H), 3,09 (dd, J = 4,8, 11,6 Hz, 1 H), 2,77 (dd, J = 4,0, 14,0 Hz, 1 H), 0,92 (s, 3 H), 0,85 (s, 3 H), 0,82 (s, 3 H), 0,72 (s, 3 H), 0,63 (s, 3 H), 0,59 (s, 3 H), 0,40 (s, 3 H) ;

[0124] 13C NMR (150 MHz, CDCl3 ) δ 175.9; 129.8, 129.6, 129.2, 128.9; 73.0, 72.9, 72.4, 70.5, 69.5, 69.0, 66.7, 62.9, 55.5, 47.6, 47.0, 45.9, 41.7, 41.1, 39.1, 38.9, 38.5, 36.7, 33.2, 32.0, 31.9, 30.7, 30.0, 29.9, 29.8, 29.7, 29.5, 29.4, 27.9, 27.8, 27.4, 26.0, 25.6, 23.6, 23.5, 22.8, 18.1. 16.6, 16.4, 15.3, 14.3 ;

[0125] HRMS (ESI) m / z calculated for C91H96O19Na [M + Na]+ 1515.6444, found 1515.6425.

[0126] 2.2 Synthesis of 28-O-2,3,4,6-tetra-O-benzoyl-β-D-glucopyranosyl oleanate 3-O- 2,3-di-O-benzoyl-β-D-glucopyranoside12 To a solution of compound 11 (5.3 g, 3.55 mmol) in a mixture of CH₂Cl₂ and MeOH (1.5:1, v / v, 2.5 mL) at 40 °C, p-TsOH-H₂O (203 mg, 1.07 mmol) was added under argon. After being stirred at the same temperature overnight, TLC showed complete conversion of the starting material. The reaction mixture was then quenched with Et₃N and concentrated under vacuum. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate: 15:1 → 6:1) to obtain compound 12 (4.23 g, 85%) as a white solid.

[0127] [α] 25 D = +92.4 (c = 0.45, CHCl3).

[0128] 1H RMN (400 MHz, CDCl3) δ 8.03-7,80 (m, 12 H), 7,55-7,25 (m, 18 H), 5,99-5,92 (m, 2 H), 5,72 (t-like, J = 9,2 Hz, 2 H), 5,48-5,37 (m, 2 H), 5,26 (br s, 1 H), 4,76 (d, J = 7,6 Hz, 1 H), 4,54 (dd, J = 2,8, 12,0 Hz, 1 H), 4,46 (dd, J = 4,2, 12,4 Hz, 1 H), 4,27-4,23 (m, 1 H), 3,99-3,85 (m, 3 H), 3.59-3.55 (m, 1 H), 3.29 (d, J = 4.0 Hz, 1 H), 3.08 (dd, J = 4.8, 11.2 Hz, 1 H), 2,77 (dd, J = 3,6, 12,4 Hz, 1 H), 0,93 (s, 3 H), 0,85 (s, 3 H), 0,81 (s, 3 H), 0,71 (s, 3 H), 0,64 (s, 3 H), 0,59 (s, 3 H), 0,40 (s, 3 H) ;

[0129] 13 C RMN (100 MHz, CDCl3 ) δ 175,9, 167.8, 166.3, 165.8, 165.3, 165.2, 164.9, 143.1, 133.8, 133.7, 133.5, 133.4, 133.3, 130.2, 130.1, 130.0, 129.9, 129.8, 129.6, 129.0, 128.8, 128.6, 128.5, 128.4, 122.9, 103.2, 92.0, 90.6, 75.8, 73.0, 71.9, 70.5, 70.4, 69.4, 62.9, 62.7, 55.5, 47.6, 47.0, 45.9, 41.7, 41.1, 39.0, 38.8, 38.5, 36.7, 33.9, 33.2, 31.9, 30.7, 29.9, 29.5, 27.9, 26.2, 25.6, 23,6, 23,5, 22,9, 22,8, 18,2, 16,6, 16,4, 15,3 ;

[0130] HRMS (ESI) m / z calculated for C84H92O19Na [M + Na]+ 1427.6131, found 1427.6191.

[0131] 2.3 Synthesis of 28-O-2,3,4,6-tetra-O-benzoyl-β-D-glucopyranosyl oleanate 3-O-2,3-di-O-benzoyl-β-D-glucuronide13 To a solution of compound 12 (3.83 g, 2.73 mmol) in a mixture of CH₂Cl₂ and H₂O (2:1, v / v, 12 mL) at room temperature, TEMPO (85.2 mg, 0.55 mmol) and BAIB (2.64 g, 8.19 mmol) were added under argon. After being stirred at room temperature for 3 h, TLC indicated complete conversion of the starting material. The reaction mixture was then quenched with saturating Na₂SO₃ and extracted with CH₂Cl₂. The combined organic layers were washed with saturating NaCl, dried over Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (CH Cl / MeOH: 30 / 1→8 / 1) to obtain 13(3.37 g, 87%) as a white solid.

[0132] [α]25 D = +64.2 (c = 0,43, CHCl3 ) ;

[0133] 1 H RMN (400 MHz, CDCl3) δ 8,02-7,80 (m, 12 H), 7,55-7,25 (m, 18 H), 5.99-5.92 (m, 2 H), 5.75-5.70 (m, 2 H), 5.55 (t, J = 8.8 Hz, 1 H), 5.48 (t, J = 9,6 Hz, 1 H), 5,26 (br s, 1 H), 4,82 (d, J = 7,6 Hz, 1 H), 4,54 (dd, J = 2,8, 12,0 Hz, 1 H), 4.46 (dd, J = 4.8, 12.4 Hz, 1 H), 4.27-4.23 (m, 1 H), 4.17-4.09 (m, 2 H), 3.11 (dd, J = 2,0, 9,6 Hz, 1 H), 2,77 (dd, J = 2,4, 12,0 Hz, 1 H), 0,92 (s, 3 H), 0,85 (s, 3 H), 0,82 (s, 3 H), 0,71 (s, 3 H), 0,64 (s, 3 H), 0,59 (s, 3 H), 0,40 (s, 3 H) ;

[0134] 13C NMR (150 MHz, CDCl3) δ 176.0, 166.3, 165.8, 165.3, 165.2, 164.9, 143.1, 133.7, 133.5, 133.4, 133.3, 130.1, 130.0, 129.9, 129.8, 129.4, 129.1, 128.8, 128.6, 128.5, 128.4, 122.9, 103.2, 92.1, 91.1, 75.1, 73.6, 73.0, 71.5, 70.6, 70.5, 69.5, 62.9, 55.4, 47.6, 47.0, 45.9, 41.7, 41.1, 39.0, 38.8, 38.5, 36.7, 33.9, 33.2, 32.1, 32.0, 31.9, 30.7, 29.9, 29.5, 27.9, 27.8, 25.9, 25.6, 23.6, 23.5, 22.9, 22.8, 18.1, 16.6, 16.4, 15.3, 14.3;

[0135] HRMS (ESI) m / z calculated for C84H89O20 [M - H]- 1417.5947, found 1417.6003.

[0136] 2.4 Synthesis of 28-O-β-D-glucopyranosyl oleanate2 To a solution of compound 10 (51 mg, 0.05 mmol) in a mixture of CH2Cl2 and MeOH (2:1, v / v, 1.5 mL) at room temperature, sodium methylate (27 mg, 0.5 mmol) was added under argon. After stirring at room temperature for 24 h, TLC showed complete conversion of the starting material. The reaction was then neutralized with Amberlite IR120 H+ resin. The mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol: 40:1 → 5:1) to obtain compound 2 (26.2 mg, 85%) as a white solid.

[0137] 1H NMR (400 MHz, pyridine-d5 ) δ 6.34 (d, J = 8.0 Hz, 1 H), 5.45 (br s, 1 H), 4.49-4.35 (m, 3 H), 4.30 (t, J = 8.8 Hz, 1 H), 4, d, 2, J = 8.8 Hz, 4.08-4.03 (m, 1 H), 3.43 (dd, J = 5.6, 10.0 Hz, 1 H), 3.21 (dd, J = 3.6, 13.6 Hz, 1 H), 1.24 (s, 3 H), H), 0.91 (s, 3 H), 0.90 (s, 3 H), 0.88 (s, 3 H);

[0138] HRMS (ESI) m / z calculated for C36H58O8 Na [M + Na]+ 641.4029, found 641.4025.

[0139] 2.5 Synthesis of 28-O-β-D-glucopyranosyl oleanate 3-O-β-D-glucopyranosideK5 To a solution of compound 12 (10 mg, 0.0071 mmol) in a mixture of dichloromethane and MeOH (2:1, v / v, 0.9 mL) at room temperature, sodium methylate (4 mg, 0.07 mmol) was added under argon. After stirring at room temperature for 24 h, TLC showed complete conversion of the starting material. The reaction was then neutralized with Amberlite IR120 H+ resin. The mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol: 40:1 → 5:1) to obtain K5 (5.0 mg, 91%) as a white solid.

[0140] 1H NMR (400 MHz, pyridine-d5) δ 6.33 (d, J = 8.0 Hz, 1 H), 5.43 (br s, 1 H), 4.94 (d, J = 7.6 Hz, 1 H), 4.59 (dd, J = 2.4, 12.0 Hz, 1 H), 4.48-4.35 (m, 4 H), 4.31-4.19 (m, 4 H), 4.07-4.02 (m, 3 H), 3.38 (dd, J = 4.0, 11.6 Hz, 1 H), 3.19 (dd, J = 2.8, 12.8 Hz, 1 H), 1.31 (s, 3 H), 1.26 (s, 3 H), 1.10 (s, 3H), 0.99 (s, 3H), 0.90 (s, 3H), 0.88 (s, 3H), 0.83 (s, 3H);

[0141] HRMS (ESI) m / z calculated for C42H68O13Na [M + Na]+ 803.4558, found 803.4564.

[0142] 3. Synthesis of compound K1

[0143] Compound 1 has the following formula:

[0144] The summary diagram is as follows:

[0145] 3.1. benzyl synthesis (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4 ,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate3:

[0146] Benzyl bromide (1.3 g, 8.55 mmol), K₂CO₃ (1.92 g, 13.14 mmol), and tetrabutylammonium bromide (0.45 g, 1.3 mmol) were added to a stirred solution of oleanolic acid (1) (3.0 g, 6.570 mmol) in THF / H₂O (10:1, 40 mL) at room temperature. The resulting reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was diluted with ethyl acetate (150 mL) and washed with water (100 mL) followed by brine solution (50 mL). The aqueous layers were again extracted with ethyl acetate (50 mL) and the combined organic layers were dried with anhydrous sodium sulfate, filtered and concentrated on the rotary evaporator.The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) eluted with 5% Ethyl Acetate in hexane to obtain benzyl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate3(3.0 g, 83%) as an off-white solid.

[0147] 3.2. synthesis of (3R,4S,5S,6S)-2-(((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-((benzyloxy)carbonyl)-4,4,6a,6b,11,11,14b-heptamethyl-1,2,3,4,4 a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate5:

[0148] On activated molecular sieves (2.0 g), benzyl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate3 (2.0 g, 3.65 mmol), (2R,3R,4S,5S,6S)-2-bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate4 (3.60 g, 9.12 mmol) and silver carbonate (1.0 g, 9.12 mmol) were added under argon, followed by dry dichloromethane (30 mL). The resulting reaction mixture was stirred at room temperature for 16 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was filtered through Celite buffer and rinsed with dichloromethane (30 mL). The filtrate was washed with water (50 mL) followed by brine solution (50 mL). The separated aqueous layers were again extracted with dichloromethane (50 mL), and the combined organic layers were concentrated on a rotary evaporator.The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) with an eluent composed of 15% ethyl acetate in hexane to obtain (3R,4S,5S,6S)-2-((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-((benzyloxy)carbonyl)-4,4,6a,6b,11,11,14b-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (5) (450 mg, 15%) in the form of an off-white solid.

[0149] 3.3. synthesis of (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-2,2,6a,6b,9,9,12a-heptamethyl-10-(((3R,4S,5S,6S)-3,4,5-triacetoxy- 6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)oxy)-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-acid octadecahydropicene-4a(2H)-carboxylic acid (6):

[0150] In a stirred solution of (3R,4S,5S,6S)-2-((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-((benzyloxy)carbonyl)-4,4,6a,6b,11,11,14b-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate5 (400 mg, 0.46 mmol) in a mixture of methanol and tetrahydrofuran (1:1; 10 mL) was added 5% W / C (50 mg). The resulting reaction mixture was stirred at room temperature under H2 flask pressure for 16 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was filtered through Celite buffer and washed with dichloromethane (30 mL) and methanol (10 mL). The filtrate was concentrated on a rotary evaporator.The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) with an eluent composed of 30% ethyl acetate in hexane to give the desired product of (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-2,2,6a,6b,9,9,12a-heptamethyl-10-(((3R,4S,5S,6S)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)oxy)-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylic6(300 mg, 84%) in the form of an off-white solid.

[0151] 3.4. Summary of (3R,4S,5S,6S)-2-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-2,2,6a,6b,9,9,12a-hept amethyl-10-(((3R,4S,5S,6S)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H- pyran-2-yl)oxy)-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicen-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate7:

[0152] In a shaken solution of (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-2,2,6a,6b,9,9,12a-heptamethyl-10-(((3R,4S,5S,6S)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)oxy)-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylic acid6 (300 mg, 0.38 mmol) in Dichloromethane:H2O (10:1, 9 mL) was added (2R,3R,4S,5S,6S)-2-bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (230 mg, 0.58 mmol), K₂CO₃ (524 mg, 3.8 mmol), tetrabutylammonium bromide (48 mg, 0.15 mmol). The resulting reaction mixture was stirred at 50 °C for 48 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was diluted with dichloromethane (100 mL) and washed with water (50 mL) followed by brine solution (50 mL). The separated aqueous layers were reconstituted with dichloromethane, and the combined organic layer was concentrated using a rotary evaporator.The resulting residue was purified by flash chromatography on a Biotage silica gel column (100-200 mesh) eluting with 50% ethyl acetate in hexane to obtain (3R,4S,5S,6S)-2-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-2,2,6a,6b,9,9,12a-hep tamethyl-10-((3R,4S,5S,6S)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H- pyran-2-yl)oxy)-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate7(300 mg, 71%) in solid form off-white.

[0153] 3.5. Summary of (2S,3S,4S,5R)-6-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(((3R,4S,5S,6S)- 6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-2,2,6a,6b,9,9,12a- heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (CompoundK1):

[0154] In a stirred solution of (3R,4S,5S,6S)-2-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-2,2,6a,6b,9,9,12a-hept amethyl-10-(((3R,4S,5S,6S)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H- pyran-2-yl)oxy)-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate7(100 mg, 0.09 mmol) in Dichloromethane:MeOH (1:1; 2 mL) was added sodium hydride (suspension at ~60% in mineral oil, 2 mg, 0.09 mmol). The resulting reaction mixture was stirred at room temperature for 3 h, then H2O (0.2 mL) was added. The progress of the reaction was monitored by TLC and LCMS. After completion of the reaction, the reaction mixture was acidified with Amberlyst 120 H+ resin, then filtered and concentrated on a rotary evaporator. The resulting residue was washed with dichloromethane (2 5 mL) et séché sous vide pour obtenir (2S,3S,4S,5R)-6-((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(((3R,4S,5S,6S)-6-carboxy-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (ComposéK1) (41.5 mg, 56%) sous forme de solide blanc cassé.

[0155] 1 H NMR (300 MHz, CDCl3) δ 13.1 - 12.1 (brs, 2 H), 6.04 - 6.01 (t, J = 6 Hz, 1H), 5.56 - 5.55 (d, J = 3 Hz, 1H), 5.33 - 5.25 (m, 1H), 5.19 - 5.12 (m, 2H), 5.04 - 5.03 (d, J = 3 Hz, 2H), 4.25 - 4.23 (d, J = 6 Hz 1H), 3.67 - 3.56 (m, 2H), 3.28 (bs, 2H), 3.16 - 2.98 (m, 5H), 2.81 - 2.76 (m, 1H), 1.91 (bs, 1H), 1.76 (bs, 2H), 1.66 - 1.47 (m, 11H), 1.38 - 1.30 (m, 4H), 1.23 - 1.07 (m, 6H), 0.97 (s, 5H), 0.86 (s, 11H), 0.67 - 0.65 (d, J = 6 Hz, 6H).

[0156] LCMS : 809.52 [M+H], 826.58 [M+NH4]

[0157] Pureté HPLC : 95,51%.

[0158] 4. Synthesis of compound K2

[0159] Compound K2 has the following formula

[0160] The summary diagram is as follows:

[0161] 4.1 Summary of (3R,4S,5S,6S)-2-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b, 7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicen-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate8:

[0162] To a stirred solution of oleanolic acid (2.0 g, 4.37 mmol) in dichloromethane / H₂O (10:1, 40 mL), (2R,3R,4S,5S,6S)-2-bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (2.25 g, 5.69 mmol) was added, followed by K₂CO₃ (1.507 g, 10.92 mmol) and Bu₄NBr (280 mg, 0.869 mmol). The mixture was refluxed under a nitrogen atmosphere. After completion (thin-layer chromatography), the reaction mixture was diluted with dichloromethane (100 mL) and washed with water (50 mL) and brine solution (50 mL). The separated aqueous layers were extracted again in Ethyl Acetate (50 mL) and the combined organic layer was dried on Na2SO4, filtered and concentrated on a rotary evaporator.The residue obtained was purified by flash chromatography on a Biotage silica gel column (100-200 mesh) eluting with 30% ethyl acetate in hexane to give the desired product of (3R,4S,5S,6S)-2-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (8(1.50 g, 44%) in the form of a white solid.

[0163] 4.2. Summary of (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-(((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-4 ,4,6a,6b,11,11,14b-heptamethyl-8a-((((3R,4S,5S,6S)-3,4,5-triacetoxy-6-(methoxyc arbonyl)tetrahydro-2H-pyran-2-yl)oxy)carbonyl)-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl tribenzoate14:

[0164] In a stirred solution of (3R,4S,5S,6S)-2-((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate5 (1.0 g, 1.293 mmol) in anhydrous dichloromethane (60 mL) at room temperature, the following was added: (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-(2,2,2-trichloro-1-iminoethoxy)tetrahydro-2H-pyran-3,4,5-triyl tribenzoate (Int-4 – synthesis described below) (1.435 g, 1.940 mmol) was followed by 4 Å molecular sieve powder (2.0 g). After 1 h, TMSOTf (30 mg, 0.129 mmol) was added by dissolving it in anhydrous dichloromethane (0.3 mL) to the above reaction mixture and kept at room temperature for 4 h. The reaction was stopped by adding triethylamine (1.8 mL, 12.9 mmol) diluted in dichloromethane (2 mL). The mixture was concentrated under reduced pressure.The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) eluting with 10% to 25% ethyl acetate in hexane to give the desired product of (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-(((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-4 ,4,6a,6b,11,11,14b-heptamethyl-8a-((((3R,4S,5S,6S)-3,4,5-triacetoxy-6-(methoxyc arbonyl)tetrahydro-2H-pyran-2-yl)oxy)carbonyl)-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl tribenzoate8(700 mg, 40%) in the form of a white solid.

[0165] 4.2bis synthesis of compound Int-4

[0166] The reaction scheme is as follows:

[0167] 4.2bis.a. Synthesis of (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-hydroxytetrahydro-2H-pyran-3,4,5-triyl tribenzoate2a:

[0168] To a stirred solution of (3R,4S,5R,6R)-6-((benzoyloxy)methyl)tetrahydro-2H-pyran-2,3,4,5-tetrayl tetrabenzoate (10.0 g, 14.27 mmol) in tetrahydrofuran (60 mL), MeNH2 (~2 M in THF, 15 mL) was added, and the mixture was stirred continuously at room temperature for 20 h. After completion of the starting material by TLC, the reaction mixture was diluted with ethyl acetate (100 mL) and washed with water (100 mL) followed by brine solution (50 mL). The separated aqueous layers were again extracted with ethyl acetate (50 mL), and the combined organic layer was dried over Na2SO4, filtered, and concentrated using a rotary evaporator. The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) eluting with 20% Ethyl Acetate in hexane to give the desired product of (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-hydroxytetrahydro-2H-pyran-3,4,5-triyl tribenzoate2a(5.50 g, 65%) as a white solid.

[0169] 4.2bis.b. Synthesis of (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-(2,2,2-trichloro-1-iminoethoxy)tetrahydro-2H-pyran-3,4,5-triyl tribenzoateInt-4:

[0170] To a stirred solution of (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-hydroxytetrahydro-2H-pyran-3,4,5-triyl tribenzoate (2a) (5.50 g, 9.22 mmol) in dichloromethane at 0 °C, trichloroacetonitrile (7.05 mL, 46.09 mmol) followed by DBU (0.3 mL, 1.89 mmol) was added. The resulting reaction mixture was left at room temperature for 2 hours and then concentrated under vacuum. The residue obtained was purified by flash chromatography on silica gel column (100-200 mesh) eluting with 15% ethyl acetate in hexane to give the desired product (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-(2,2,2-trichloro-1-iminoethoxy)tetrahydro-2H-pyran-3,4,5-triyl tribenzoateInt-4(5.60 g, 82%) as a white solid.

[0171] 4.3. Synthèse du (2S,3S,4S,5R)-6-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-2,2,6a,6b,9,9,12a-heptamethyl-10-(((3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (ComposéK2):

[0172] In a shaken solution of (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-4,4,6a,6b,11,11,14b-heptamethyl-8a-((((3R,4S,5S,6S)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)oxy)carbonyl)-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl Tribenzoate-14 (350 mg, 0.28 mmol) in MeOH / Dichloromethane (1:1, 10 mL) was added to NaH (~60% in mineral oil, 7 mg, 0.26 mmol) and the mixture was kept at room temperature for 16 h. The reaction mixture was then treated with water (3.5 mL) and kept at room temperature for another 6 h. The progress of the reaction was monitored by TLC and LCMS. After completion, the reaction was acidified with Amberlyst-120 H+ resin, and the mixture was filtered through Celite buffer, rinsed with MeOH / Dichloromethane (1:1, 10 mL), and the filtrate was concentrated on a rotary evaporator.The resulting residue was purified by silica gel chromatography using a gradient elution of 5% to 20% methanol in dichloromethane to yield the expected product. (2S,3S,4S,5R)-6-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-2,2,6a,6b,9,9,12a-hept amethyl-10-(((3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyr an-2-yl)oxy)-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylicK2(93 mg, 45%) as an off-white solid.

[0173] LCMS: m / z 795.529 [M+H]

[0174] HPLC Purity: 93.25%

[0175] 1H NMR (300 MHz, DMSO-d6) δ 12.80 (br s, 1H), 6.02 (t, J = 3.0 Hz, 1H), 5.52 - 5.56 (m, 1H), 5.10 - 5.32 (m, 3H), 4.82 - 4. 90 (m, 3H), 4.32 - 4.42 (m, 1H), 4.12 (d, J = 12.0 Hz, 1H), 3.90 (br s, 1H), 3.54 - 3.68 (m, 2H), 3.40 (m, 1H), 3.25 (m, 1H), 3.00 - 3.10 (m, 5H), 2.90 - 2.96 (m, 1H). 1.90 - 1.96 (m, 1H), 1.72 - 1.86 (m, 4H), 1.40 - 1.68 (m, 11H), 1.20 - 1.40 (m, 5H), 1.07 (s, 3H), 0.97 (s, 3H), 0.86 (s, 9H), 0.75 (s, 3H), 0.67 (s, 3H), 0.66 (s, 3H).

[0176] 5. synthèse du composeK3

[0177] Le composéK3a la formule suivante :

[0178] Le schéma réactionnel est le suivant :

[0179] 5.1. Synthèse de l’allyl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate15:

[0180] In a stirred solution of (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylic acid1(4.0 g, 8.6 mmol) in THF, sodium hydride (dispersion in hexane, 55% wt, 3.20 g, 136.2 mmol) was added per portion at 0 ℃. The reaction mixture was stirred at 0 °C for 3 h, then allyl bromide (5.20 g, 42.6 mmol) was added, followed by tetrabutylammonium iodide (1.60 g, 4.2 mmol). The resulting reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion, the reaction was diluted with ethyl acetate (150 mL) and washed with water (100 mL) followed by brine solution (100 mL).The separated aqueous layers were again extracted with ethyl acetate (50 mL) and the combined organic layer was dried with anhydrous sodium sulfate, filtered and concentrated on a rotary evaporator. The residue obtained was purified by flash chromatography on Biotage silica gel (100-200 mesh) by eluting with 2% ethyl acetate in hexane to obtain allyl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b octadecahydropicene-4a(2H)-carboxylate15 as an off-white solid (1.9 g, 40%).

[0181] 5.2. Summary of (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1, 3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylic acid16:

[0182] In a stirred solution of allyl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate15(100 mg, 0.18 mmol) in ethyl cetate and DCM (1:1; 8 mL) 1,3-Dimethylbarbituric acid (30 mg, 0.18 mmol) and Pd(PPh3)4(6 mg, 0.004 mmol) were added at room temperature. The resulting reaction mixture was stirred at room temperature for 1 h. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was filtered through Celite buffer and rinsed with ethyl acetate. The filtered liquid was washed with water and then with brine. The separated aqueous layer was extracted again with ethyl acetate, and the combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated using a rotary evaporator.The residue obtained was purified by Biotage silica gel column chromatography (100-200 mesh) by eluting with 10% ethyl acetate in hexane to obtain (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylic16(70 mg, 76%) as an off-white solid.

[0183] 5.3. Summary of (2R,3R,4S,5R)-2-(acetoxymethyl)-6-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl- 1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate17:

[0184] In a shaken solution of (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylic16 (750 mg, 1.52 mmol) in a dichloromethane:H2O mixture (10:1; 15 mL), (2R,3R,4S,5R,6R)-2-(acetoxymethyl)-6-bromotetrahydro-2H-pyran-3,4,5-triyl triacetate (810 mg, 1.96 mmol), potassium carbonate (626 mg, 4.5 mmol), and tetrabutyl ammonium bromide (49 mg, 0.15 mmol) at room temperature. The resulting reaction mixture was stirred at 50 °C for 16 h. The reaction progress was monitored by TLC. After the reaction was complete, the reaction mixture was extracted with dichloromethane (100 mL) and washed with water followed by brine. The separated aqueous layers were again extracted with dichloromethane, and the combined organic layers were dried over sodium sulfate, filtered, and concentrated using a rotary evaporator.The residue obtained was purified by flash chromatography on silica gel (100-200 mesh) by eluting with 15% ethyl acetate in hexane to obtain (2R,3R,4S,5R)-2-(acetoxymethyl)-6-((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (1.0 g, 80%) under off-white solid shape.

[0185] 5.4. Summary of (2R,3R,4S,5R)-2-(acetoxymethyl)-6-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptam ethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate18:

[0186] In a stirred solution of (2R,3R,4S,5R)-2-(acetoxymethyl)-6-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (400 mg, 0.48 mmol) in an acetone:H2O mixture (2:1; 12 mL), N-methylmorpholine N-oxide (74 mg, 0.62 mmol) and potassium osmate (9 mg, 0.02 mmol) at room temperature. The resulting reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion, the reaction was filtered through Celite buffer and rinsed with ethyl acetate. The filtrate was washed with water and then brine. The combined aqueous layers were extracted with ethyl acetate, and the combined organic layers were dried over sodium sulfate, filtered, and concentrated using a rotary evaporator.The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) eluting with 60% ethyl acetate in hexane to obtain (2R,3R,4S,5R)-2-(acetoxymethyl)-6-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate18(400 mg, 96%) in the form of an off-white solid.

[0187] 5.5. Synthesis of (3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptameth yl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate (CompoundK3):

[0188] In a stirred solution of (2R,3R,4S,5R)-2-(acetoxymethyl)-6-((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (180 mg, 0.2 mmol) in a dichloromethane and methanol mixture (1:1; 8 mL), sodium hydride (suspension) was added. ~55% in mineral oil, 5 mg, 0.2 mmol). The resulting reaction mixture was stirred at room temperature for 3 h. The progress of the reaction was monitored by TLC. After completion, the reaction was acidified with Amberlyst 120 H+ resin. The reaction mixture was filtered through a cotton plug, and the filtrate was concentrated on a rotary evaporator.The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) by eluting with 5% MeOH in dichloromethane to obtain (3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate (Compound K3) (75 mg, 52%) as solid off-white.

[0189] 1 H NMR (300 MHz, CDCl3) δ 5.24 - 5.22 (d, J = 3 Hz, 1H), 5.19 - 5.17 (m, 2H), 5.05 - 50.3 (d, J = 6 Hz 1H), 4.97 - 4.96 (d, J = 6 Hz, 1H), 4.52 - 4.49 (m, 1H), 4.45 - 4.40 (m, 2H), 3.64 - 3.59 (m, 1H), 3.53 - 3.38 (m, 4H), 3.21 - 3.08 (m, 5H), 2.75 - 2.72 (m, 2H), 1.99 - 1.91 (m, 1H), 1.81 - 1.78 (m, 2H), 1.68 - 1.45 (m, 9H), 1.39 - 1.22 (m, 5H), 5.83 (s, 6H), 0.93 - 0.83 (m, 15H), 0.70 - 0.68 (d, J = 6 Hz, 6H).

[0190] LCMS: 710.49 [M+NH4]

[0191] HPLC purity: 95%

[0192] 6. Synthesis of compound K4

[0193] Compound K4 has the following formula:

[0194] The summary diagram:

[0195] 6.1. Synthesis of allyl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,3, 4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate19:

[0196] In a stirred solution of (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylic acid (1) (2.0 g, 4.3 mmol) in THF (25 mL) sodium hydride (suspension at ~55% in mineral oil, 1.60 g, 68.8 mmol) was added in portions at 0 ℃. The reaction mixture was stirred at 0 °C for 3 h, then allyl bromide (2.6 g, 21.8 mmol) and tetrabutylammonium iodide (0.8 g, 2.1 mmol) were added. The resulting reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion, the reaction was diluted with ethyl acetate (150 mL) and washed with water (50 mL) followed by brine solution (50 mL).The separated aqueous layers were again extracted with ethyl acetate and the combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated on a rotary evaporator. The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) eluted with 2% Ethyl Acetate in hexane to obtain allyl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate19(900 mg, 39%) as an off-white solid.

[0197] 6.2.Synthesis of 2,3-dihydroxypropyl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptameth yl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate (CompoundK4):

[0198] In a stirred solution of allyl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate19(400 mg, 0.74 mmol) in acetone and H2O (3:1; 12 ml) N-methylmorpholine N-oxide (133 mg, 68.8 mmol) and potassium osmate (14 mg, 0.03 mmol) were added at room temperature. The resulting reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by thin-layer chromatography (TLC). After completion, the reaction was filtered through Celite buffer and rinsed with ethyl acetate (50 mL). The filtrate was washed with water (10 mL) followed by brine solution (10 mL). The separated aqueous layers were extracted again with ethyl acetate (25 mL), and the combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated using a rotary evaporator.The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) with an eluent composed of 50% ethyl acetate in hexane to obtain 2,3-dihydroxypropyl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate (Compound K4) (80 mg, 18%) as an off-white solid.

[0199] 1 H NMR (300 MHz, CDCl3) δ5.18 (bs, 1H), 4.83 - 4.81 (d,J= 6 Hz, 1H), 4.62 - 4.57 (m, 1H), 4.52 - 4.49 (m, 1H), 4.44 - 4.40 (m, 1H), 3.95 - 3.82 (m, 2H), 3.61 - 3.49 (m, 5H), 3.18 - 3.17 (m, 2H), 2.80 - 2.73 (m, 2H), 1.91 - 1.23 (m, 20H), 1.09 (s, 4H), 0.93 - 0.86 (m, 14H), 0.7 - 0.66 (d,J= 12 Hz, 6H)

[0200] LCMS :622.41 [M+NH4]

[0201] HPLC purity: 93.86%.

[0202] 7. Synthesis of compound K5

[0203] Compound K5 has the following formula

[0204] Le schéma de synthèse est le suivant :

[0205] 7.1. Synthèse de (2R,3R,4S,5R)-2-(acetoxymethyl)-6-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate20:

[0206] To a stirred solution of oleanolic acid (500 mg, 1.094 mmol) in a dichloromethane / H₂O mixture (10:1, 30 mL), (2R,3R,4S,5R,6R)-2-(acetoxymethyl)-6-bromotetrahydro-2H-pyran-3,4,5-triyl triacetate (585 mg, 1.422 mmol) was added, followed by potassium carbonate (1.89 g, 13.684 mmol) and tetrabutylammonium bromide (36 mg, 0.109 mmol). The mixture was refluxed under a nitrogen atmosphere for 6 h. After completion of the reaction (thin-layer chromatography), the reaction mixture was diluted with dichloromethane (50 mL) and washed with water (20 mL) and brine solution (20 mL). The separated aqueous layers were extracted again in dichloromethane (50 mL) and the combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated on a rotary evaporator.The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) eluted with 30% ethyl acetate in hexane to give the desired product, namely (2R,3R,4S,5R)-2-(acetoxymethyl)-6-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate20(600 mg, 70%) in the form of a white solid.

[0207] 7.2.Synthesis of (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-(((3S,4aR,6aR,6bS,8aS,12aS,14aR, 14bR)-4,4,6a,6b,11,11,14b-heptamethyl-8a-((((3R,4S,5R,6R)-3,4,5-triacetoxy-6-(acetoxy methyl)tetrahydro-2H-pyran-2-yl)oxy)carbonyl)-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyltribenzoate21:

[0208] In a shaken solution of (2R,3R,4S,5R)-2-(acetoxymethyl)-6-((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate20(500 mg, 0.635 mmol) in anhydrous dichloromethane (30 mL) at room temperature, the following was added: (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-(2,2,2-trichloro-1-iminoethoxy)tetrahydro-2H-pyran-3,4,5-triyl tribenzoate (Int-4) (705 mg, 0.952 mmol) followed by 4 Å molecular sieve powder (1.0 g). After 1 h, trimethylsilyl triflate (15 mg, 0.063 mmol) was added in dichloromethane solution anhydre (0.15 mL) au mélange réactionnel ci-dessus et la réaction a été poursuivie à température ambiante pendant 4 h. La réaction a été arrêtée par l'ajout de triéthylamine (1.8 mL, 12.9 mmol) diluée dans le dichlorométhane(2 mL). Le mélange a été concentré sous pression réduite pour donner le produit brut. Celui-ci a été purifié par chromatographie flash sur colonne de gel de silice (100-200 mesh) éluée avec 20% d'acétate d’éthyle dans l'hexane pour donner le produit désiré de (2R,3R,4S,5R)-2-((benzoyloxy)méthyl)-6-(((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-4,4,6a,6b,11,11,14b-heptaméthyl-8a-((((3R,4S,5R,6R)-3,4,5-triacétoxy-6-(acétoxyméthyl)tétrahydro-2H-pyran-2-yl)oxy)carbonyl)-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)tétrahydro-2H-pyran-3,4,5-triyl tribenzoate21(270 mg, 31%) sous forme de solide blanc.

[0209] 7.3.Synthèse de (3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxyméthyl)tétrahydro-2H-pyran-2-yl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-2,2,6a,6b,9,9,12a-heptaméthyl-10-(((3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate (ComposéK5):

[0210] In a shaken solution of (2R,3R,4S,5R)-2-((benzoyloxy)methyl)-6-((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-4,4,6a,6b,11,11,14b-heptamethyl-8a-((((3R,4S,5R,6R)-3,4,5-triacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)oxy)carbonyl)-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl Tribenzoate 21 (260 mg, 0.191 mmol) in a methanol / dichloromethane mixture (1:1, 8 mL) was mixed with sodium hydride (dispersed at ~60% in mineral oil, 8 mg, 0.191 mmol), and the reaction continued at room temperature for 4 h. The reaction mixture was acidified with Amberlyst-120 H resin. + and the mixture was filtered through a Celite buffer, rinsed with a methanol / dichloromethane mixture (1:1, 10 mL) et le filtrat a été concentré à l’évaporateur rotatif pour donner le produit brut. Ce dernier a été purifié par chromatographie flash sur colonne de gel de silice (100-200 mesh) en éluant avec 5% de MeOH dans le dichlorométhanepour donner le produit souhaité de (3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxyméthyl)tétrahydro-2H-pyran-2-yl (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-2,2,6a,6b,9,9,12a-heptamethyl-10-(((3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadécahydropicène-4a(2H)-carboxylate (ComposéK5) (116 mg, 77%) sous forme de solide blanc.

[0211] 1H NMR(300 MHz,DMSO-d6) δ 5.16 - 5.24 (m, 2H), 5.10 (d,J= 6.0 Hz, 1H), 5.04 (d,J= 6.0 Hz, 1H), 4.95 (d,J= 6.0 Hz, 1H), 4.92 (d,J= 6.0 Hz, 1H), 4.88 (d,J= 6.0 Hz, 1H), 4.49 (t,J= 6.0 Hz, 1H), 4.41 (t,J= 6.0 Hz, 1H), 4.12 - 4.16 (m, 1H), 3.52 - 3.68 (m, 2H), 3.38 - 3.46 (m, 2H), 2.90 - 3.26 (m, 10H), 2.72 - 2.76 (m, 1H), 1.40 - 2.01 (m, 12H), 1.10 - 1.40 (m, 5H), 1.07 (s, 3H), 0.97 (s, 3H), 0.87 (s, 6H), 0.86 (s, 3H), 0.74 (s, 3H), 0.68 (s, 3H).

[0212] LCMS :m / z781.53 [M+H]

[0213] Pureté HPLC :97,4%

[0214] 8. Synthèse ducomposéK6

[0215] Le composeK6a la formule suivante

[0216] Le schéma de synthèse est le suivant :

[0217] 8.1.Synthèsedel’allyl(4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate22:

[0218] To a stirred solution of oleanolic acid (1) (2.0 g, 4.386 mmol, 1 eq) in THF / H₂O (10:1; 22 mL), allyl bromide (689 mg, 5.701 mmol), anhydrous potassium carbonate (1.210 g, 8.772 mmol), and tetrabutylammonium bromide (282 mg, 0.877) were added at room temperature. The resulting reaction mixture was stirred at 40 °C for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (150 mL) and washed with water (50 mL). The separated organic layer was dried over Na₂O. 2SO4anhydre, filtrée et concentrée sous pression réduite. Le brut a été purifié par chromatographie flash sur colonne de gel de silice (100-200 mesh) avec pour éluant un mélange composé de 10% d'acétate d’éthyle dans l'hexane pour obtenir l'allyle (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate22(2.0 g, 95 %) sous forme de solide blanc.

[0219] 8.2.Synthesis of ((3R,4S,5S,6S)-2-((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-((allyloxy)carbonyl)-4,4,6a,6b,11,11,14b-heptamethyl- 1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxyl)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate23:

[0220] Allyl(4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate22(3.0 g, 6.224 mmol) and (2R,3R,4S,5S,6S)-2-bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate4(3.706 g, 9.336 mmol) in dry dichloromethane solution (38 mL) were treated with carbonate under argon. of silver (2.574, 9.336 mmol) and 4Å molecular sieve. The resulting reaction mixture was stirred in the dark for 48 hours at room temperature. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was filtered, and the filtrate was diluted with dichloromethane (150 mL) and washed with water (100 mL). The separated organic layer was dried over Na 2SO4anhydre, filtrée et concentrée sur un évaporateur rotatif pour donner le produit brut, qui a été purifié par chromatographie flash sur colonne de gel de silice (100-200 mesh) avec un éluant composé de 10% d'Acétate d’éthyle dans de l'hexane pour obtenir (3R,4S,5S,6S)-2-(((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-((allyloxy)carbonyl)-4,4,6a,6b,11,11,14b-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14ab-icosahydropicen-3-yl)oxy)-6-(méthoxycarbonyl)tétrahydro-2H-pyran-3,4,5-triyl triacétate23(1.0 g, 20 %) sous forme de solide blanc cassé.

[0221] 8.3.Synthesis of (3R,4S,5S,6S)-2-((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-((2,3-dihydroxypropoxy)carbonyl)-4,4,6a,6b,11,11,14b-heptamethyl-1,2, 3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate24:

[0222] In a shaken solution of (3R,4S,5S,6S)-2-((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-((allyloxy)carbonyl)-4,4,6a,6b,11,11,14b-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate23(500 mg, 0.618 mmol) in acetone and H2O (1:0.5; 7.5 mL), the following was added: N-methylmorpholine-N-oxide (120 mg, 1.980 mmol), then potassium osmate K2OsO4 (12 mg, 0.031 mmol). The resulting reaction mixture was stirred at room temperature for 16 h. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (50 mL) and washed with water (20 mL) followed by brine solution (20 mL). The separated aqueous layers were again extracted with ethyl acetate (25 mL), and the combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated on a rotary evaporator to give a crude compound.The crude compound was purified by flash chromatography on silica gel (100-200 mesh) with an eluent composed of 30% ethyl acetate in hexane to give (3R,4S,5S,6S)-2-((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-((2,3-dihydroxypropoxy)carbonyl)-4,4,6a,6b,11,11,14b-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate24(180 mg, 57%) as an off-white solid.

[0223] 8.4.Synthesis of (3R,4S,5S,6S)-2-(((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-((2,3-dihydroxypropoxy)carbonyl)-4,4,6a,6b,11,11,14b-heptamethyl-1,2, 3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (CompoundK6):

[0224] In a shaken solution of (3R,4S,5S,6S)-2-((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-((2,3-dihydroxypropoxy)carbonyl)-4,4,6a,6b,11,11,14b-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate24(180 mg, 0.213 mmol) in a mixture of dichloromethane and methanol (1:1, 10 mL) was added Sodium hydride (55% suspension in mineral oil, 6 mg, 0.213 mmol) was used. The resulting reaction mixture was stirred at room temperature for 3 h, then water (5 mL) was added and stirring continued for another 16 h. After the reaction was complete, the reaction mixture was acidified with Amberlyst H resin. + The reaction mixture was filtered through a Celite bed and rinsed with a dichloromethane mixture. et de méthanol (1 :1, 20 mL), le filtrat a été concentré sur un évaporateur rotatif pour donner le produit brut. Le solide résultant a été trituré avec CH2Cl2puis lavé avec du pentane pour donner le produit désiré de (3R,4S,5S,6S)-2-((3S,4aR,6aR,6bS,8aS,12aS,14aR,14bR)-8a-((2,3-dihydroxypropoxy)carbonyl)-4,4,6a,6b,11,11,14b-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-6-(méthoxycarbonyl)tétrahydro-2H-pyran-3,4,5-triyl triacétate (ComposéK6) sous forme de solide blanc cassé (110 mg, 73%).

[0225] 1H NMR (300 MHz,DMSO-d6) δ: 12.60 (brs, 1H), 5.18 (brs, 1H), 5.04-5.03 (d, 2H,J=3Hz), 4.83-4.81 (d, 1H,J=6Hz), 4.62-4.57 (m, 1H), 4.25-4.23 (d, 1H,J=6Hz), 3.99-3.78 (m, 2H), 3.58-3.55 (m, 2H), 3.40-3.25 (m, 2H), 3.25-2.95 (m, 3H), 2.79-2.76 (m, 1H), 1.99-1.91 (m, 1H), 1.80-1.78 (m, 2H), 1.63-1.48 (m, 10H), 1.37-1.29 (m, 3H), 1.23-0.85 (m, 23H), 0.75 (s, 3H), 0.65 (s, 3H).

[0226] LCMS: 707.45 [M+H +

[0227] HPLC purity: 96%.

[0228] 9. Synthesis of compound K7

[0229] The compound K7a has the formula

[0230] The summary diagram is as follows

[0231] 9.1. Summary of (3R,4S,5S,6S)-2-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6 b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicen-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate25:

[0232] In a shaken solution of (4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylic16 (500 mg, 1.01 mmol) in a mixture of dichloromethane and water (10:1, 30 mL) was added (2R,3R,4S,5S,6S)-2-bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate4 (544 mg, 1.31 mmol) followed by anhydrous potassium carbonate (1.670 g, 13.68 mmol) and tetrabutylammonium bromide (65 mg, 0.2 mmol). The mixture was refluxed under a nitrogen atmosphere for 8 h. After completion (TLC), the reaction mixture was diluted with dichloromethane (50 mL) and washed with water (50 mL) and brine solution (50 mL). The separated aqueous layers were again extracted with dichloromethane (50 mL), and the combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated on a rotary evaporator.The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) with a 15% solution of ethyl acetate in hexane as the eluent to give the desired product of (3R,4S,5S,6S)-2-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate25(400 mg, 49%) in the form of a white solid.

[0233] 9.2.Synthesis of (3R,4S,5S,6S)-2-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b ,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicen-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate26:

[0234] In a shaken solution of (3R,4S,5S,6S)-2-((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(allyloxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate25(500 mg, 0.615 mmol) in acetone / water (2:1, 7.5 ml) potassium osmate (12 mg, 0.031 mmol) followed by N-methylmorpholine-N-oxide (115 mg, 0.984 mmol). After 16 hours of stirring at room temperature, the reaction mixture was filtered through Celite buffer and rinsed with acetone (20 mL). The filtrate was concentrated on a rotary evaporator.The residue obtained was purified by flash chromatography on a silica gel column (100-200 mesh) with a 40% ethyl acetate in hexane solution as the eluent to give the desired product of (3R,4S,5S,6S)-2-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate26(300 mg, 57%) in the form of a white solid.

[0235] 9.3.Synthesis of (2S,3S,4S,5R)-6-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3, 4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicen-4a-carbonyl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (CompoundK7):

[0236] In a stirred solution of (3R,4S,5S,6S)-2-((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate26 (100 mg, 0.118 mmol) in a methanol / dichloromethane mixture (1:1, 6 mL), sodium hydride (suspension at ~60% in mineral oil (10 mg, 0.25 mmol) and maintained at room temperature for 3 h. The reaction mixture was treated with water (1 mL) and maintained at room temperature for 48 h. The reaction was acidified with Amberlyst 120 H + and the mixture was filtered through a Celite buffer, washed with a methanol / dichloromethane solution (1:1, 10 mL) et le filtrat a été concentré sur un évaporateur rotatif pour donner le produit brut. Ce dernier a été purifié par trituration avec du dichlorométhaneet précipité dans du pentane ; le solide a été filtré et séché pour donner le produit souhaité de (2S,3S,4S,5R)-6-(((4aS,6aS,6bR,8aR,10S,12aR,12bR,14bS)-10-(2,3-dihydroxypropoxy)-2,2,6a,6b,9,9,12a-heptaméthyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicène-4a-carbonyl)oxy)-3,4,5-trihydroxytétrahydro-2H-pyran-2-carboxylique (ComposéK7) (81 mg, 64%) sous la forme d'un solide blanc cassé.

[0237] 1H NMR (300 MHz,DMSO-d6)δ 12.6 (br s, 1H), 6.02 (dd,J= 6.0 Hz, 1H), 5.55 (d,J= 3.0 Hz, 1H), 5.24 - 5.31 (m, 1H), 5.13 - 5.18 (m, 2H), 4.50 - 4.52 (m, 1H), 4.42 (t,J= 6.0 Hz, 1H), 3.93 (m, 1H), 3.49 - 3.61 (m, 1H), 3.38 - 3.42 (m, 3H), 3.16 - 3.28 (m, 3H), 2.73 - 2.77 (m, 2H), 1.91 - 1.96 (m, 1H), 1.78 - 1.81 (m, 2H), 1.44 - 1.70 (m, 9H), 1.23 - 1.39 (m, 5H), 1.08 (s, 6H), 0.92 - 0.93 (m, 15H), 0.66 - 0.70 (m, 6H).

[0238] LCMS :m / z706.50 [M+ H]

[0239] HPLC purity: 90.11% (combined purity) Example 2: Study of depigmenting properties

[0240] The aim of this study was to evaluate the potential depigmenting effects of compounds by measuring melanin production in normal human epidermal melanocytes (NHEM). The effect of the tested compound on melanin synthesis in moderately pigmented NHEMMP cells stimulated by a mixture of L-tyrosine and α-MSH was investigated using photometry.

[0241] MATERIALS AND METHODS

[0242] Biological model

[0243] - Cell type: Moderately pigmented normal human epidermal melanocytes (NHEM-MP), Bioalternatives reference: NHEM-MP used in the 11th pass

[0244] - Growing conditions: 37°C, 5% CO2

[0245] - Culture medium: Dermalife base medium optimized for the test, supplemented with

[0246] Dermalife M Lifefactors®

[0247] - Test environment:

[0248] - M254 base medium optimized for testing, supplemented with HMGS-2 without PMA

[0249] Compounds, reference and inductors

[0250] The compounds shown in Table 2 were tested:

[0251] Tested compound Appearance / Storage Stock solution Tested concentrations Compound C1 Calenduloside FES221211-10 - Powder - Storage: Room temperature (TA) 30 mM in DMSO 0.25, 2.5 and 25 µM Lipoic acid Sigma-Aldrich, ref. T1395 10 mg / ml in ethanol 5 µg / ml L-tyrosine Sigma-Aldrich, ref. T1145 250 mM in water 300 µM + 0.1 µM α-MS Sigma-Aldrich, ref. M4135 0.6 mM in water 300 µM + 0.1 µM

[0252] Cultures and treatments

[0253] Melanocytes were seeded in 24-well plates and cultured in culture medium for 24 hours. The medium was then replaced with test medium containing or not (control) the compounds or the reference (described in § 2.2), and the cells were then pre-incubated for 24 hours. After pre-incubation, the medium was replaced with test medium containing or not (control) the compounds or the reference, and the inducer, a mixture of L-tyrosine and α-MSH, was added. The cells were incubated for 9 days, with repeat treatments and stimulation after 2 and 6 days of incubation. An unstimulated control condition was performed concurrently.

[0254] All experimental conditions were carried out in n=3.

[0255] Melanin dosage

[0256] At the end of incubation, melanin was extracted by cell lysis with a 0.5 N NaOH solution. The optical density (OD) of the samples was measured at 405 nm, and then the amount of melanin was determined by comparison with a range of exogenous melanin (melanin curve including standards from 0.39 to 100 µg / ml). The results were expressed as µg / ml of melanin.

[0257] Data processing

[0258] The raw data was transferred and processed using Microsoft Excel® software.

[0259] Intergroup comparisons were performed using the unpaired two-tailed Student's t-test. Statistical analyses can be interpreted if n ≥ 5; however, for n < 5, the calculated data are provided for illustrative purposes only.

[0260] Formula used:

[0261] Standard error of the mean: esm = standard deviation (Sd) / √n

[0262] The standard error of the mean (SEM) represents the deviation of the mean from

[0263] the sample relative to the true population mean. The esm is calculated by dividing the Sd (standard deviation) by the square root of the sample size.

[0264] RESULTS

[0265] Treatment of melanocytes (NHEM) with the L-Tyrosine + α-MSH mixture resulted in an increase in melanin synthesis from 8 µg / ml to 21.8 µg / ml (mean of plates 1 and 2). This effect was inhibited by the reference compound, lipoic acid, with melanin concentration reduced to approximately 23% compared to the stimulated control (mean of plates 1 and 2). These results were expected and validated this trial.

[0266] Under the experimental conditions of this study, the compounds showed the following effects:

[0267] Compound 1: Tested at 25 µM, the compound had a cytotoxic effect on melanocytes. Tested at 0.25 and 2.5 µM, this compound showed a marked inhibitory effect on melanin synthesis (56% and 47% of the stimulated control, respectively).

[0268] The data are referenced in Table 3 below.

[0269] Compounds tested Concentration Melanin (µg / ml) Mean (µg / ml) esm (µg / ml) % Stimulated control m(%) p (1) Unstimulated control -8.0 8.28 28.10 1380***Stimulated conditions: L-Tyrosine 300 µM + aMSH 0.1 µM Control -20.3 22.2 21.3 21.3 0.5 100 3-Lipoic acid 5 µg / ml 4.9 4.8 4.6 4.8 0.1 220***Compound C 10.1 µM 22.1 21.5 21.6 21.7 0.29 71 ns 0.5 µM 15.9 9 15.4 15.2 15.5 0.26 91***2.5 µM3.2 3.23.23.20.0140***

[0270] CONCLUSION

[0271] The results obtained in this study showed that compound C1 has interesting depigmentation properties on reconstituted in vivo skin models.

[0272] Example 3: Study of depigmenting properties evaluated on human explants

[0273] The objective of this study was to evaluate the effect of the compounds according to the invention, administered systemically, on skin pigmentation by analyzing melanin content and the melanogenesis process in NativeSkin models exposed to UVA+UVB.

[0274] NativeSkin is a standardized, ready-to-use ex vivo human skin model produced from samples of consenting donors, enabling the prediction of biological responses in humans.

[0275] This model consists of 8 to 23 mm biopsies embedded in a gel matrix in the presence of epidermis and in contact with air. The system is mounted in a cell culture insert and then in a multi-well plate, allowing experiments to be carried out over more than 7 days.

[0276] Materials and methods

[0277] 16 NativeSkin models with an internal diameter of 11 mm were produced from a 48-year-old donor, having a phototype of type 3.

[0278] The models were cultured at 37°C, 5% CO2, in a saturated humidity atmosphere, with 1 mL of NativeSkin culture medium, renewed daily.

[0279] From day 0 to day 6, skin models were systemically treated with compound I described in Example 2. On days 1 to 6, the skin models were exposed to 25 mJ / cm² 2 of UVB and at 2.25 J / cm² 2 UVA was used with the Opsytec Dr.Gröbe BS-02 UV chamber. Systemic treatment was maintained in the medium throughout the duration of the culture and UVA / UVB irradiations.

[0280] Custom silicone rings were placed on the matrix to protect its integrity during UV exposure and were subsequently removed. The culture medium was renewed after UV exposure.

[0281] On day 0 (untreated control) and day 7, the skin models were carefully removed from the molds. The skin biopsies were re-penetrated to remove the area under the silicone ring, split in half, and fixed in 10% buffered formalin, then processed for embedding in paraffin wax.

[0282] The experimental design is presented below in Table 3:

[0283] #TreatmentExposureUVAnalysisTreatmentDoseRouteFreqDoseFreqJ0J71- (control)N / YR=1N =32UVA+UVBN / A2.25J / cm 2 UVA+25mJ / cm2 UVBQ(J1 to 6)-N =33Compound C12 µMSQ(J1 to 6)-N =340.5 µM-N =350.125 µM-N =3

[0284] S: systemic, Q: daily, #: test

[0285] Histological analyses

[0286] Hematoxylin and eosin (H&E) staining was performed on 5 µm thick paraffin-embedded skin cross-sections to analyze structural integrity and skin viability. Three representative images of the epidermis / dermis were acquired at 40x magnification using a Leica DMi1 microscope.

[0287] Tyrosinase immunostaining was performed on paraffin-embedded 5 µm thick skin cross-sections (see Table III for staining conditions) to analyze melanocytes in the epidermis. A 20x magnification slide scan was acquired and used for signal quantification. Representative images of the epidermis were taken at 40x magnification. The antibody used was a rabbit anti-tyrosinase antibody (ab180753) at a concentration of 3.99 mg / mL, used at a dilution of 1 / 62.5. Staining was detected with a goat anti-rabbit antibody coupled to Alexa Fluor 647.

[0288] Fontana-Masson staining for skin melanin content was performed on 5 μm thick cross-sectional skin sections. A 20x magnification slide scan was acquired and used for signal quantification. Representative images of the epidermis were taken at 40x magnification.

[0289] Image quantification

[0290] Tyrosinase quantification was performed using Quantacell. A slide scan image of an entire section of the sample was quantified. For each scan, the mean fluorescence intensity (MFI) and the percentage of positive cells in the total epidermis were quantified.

[0291] For each condition, individual values, the mean, and the SEM were plotted using GraphPad Prism. Statistical analysis was performed using a one-way ANOVA test.

[0292] Fontana-Masson quantification was performed using Quantacell. An entire section of the sample was quantified by slide scanning. For each scan, the mean staining intensity and the area of ​​the positive spot in the total epidermis, as well as in the basal layer of the epidermis where melanocytes are located, were quantified.

[0293] For each condition, individual values, the mean, and the SEM were plotted using GraphPad Prism. Statistical analysis was performed using a one-way ANOVA test.

[0294] Results and conclusions

[0295] 1- Evaluation of skin structure and integrity by H&E staining on days 0 and 7.

[0296] H&E staining of the skin model collected on day 0 revealed, as expected, histologically healthy skin with no signs of impaired viability and / or structural integrity. H&E staining of the skin models on day 7 revealed that the UVA+UVB exposure doses did not result in significant changes or abnormalities compared to untreated controls. Cells remained viable, and tissue integrity appeared normal, as expected. Only a few pyknotic cells were observed in the UVA+UVB-exposed skin models. However, the overall results suggest that UVA+UVB-treated skin models would not undergo impaired melanogenesis.

[0297] H&E staining of skin models exposed to UVA+UVB and treated with compound 1 revealed no perceptible signs of toxicity or loss of viability, compared to skin models exposed to UVA+UVB but not treated with compound 1. This confirms that, under these conditions, compound 1 does not induce any toxicity in ex vivo human skin.

[0298] 2- Evaluation of tyrosinase expression on day 7

[0299] Tyrosinase immunofluorescence staining reveals that tyrosinase expression is induced, as expected, by UVA+UVB exposure in the basal and suprabasal layers of the epidermis compared to untreated skin models ( ). Furthermore, statistical analysis confirms a significant increase in tyrosinase expression compared to untreated controls ( ).

[0300] It is interesting to note that compound 1, added to the culture medium of treated skin models, inhibits UVA+UVB induced tyrosinase expression in the epidermis, compared to UVA+UVB treated skin controls ( ). In particular, the reduction in tyrosinase expression is statistically significant for skin models treated with compound 1 at 0.125 µM compared to models exposed to UVA+UVB ( ).

[0301] Finally, under the tested conditions, no dose-dependent inhibition of UVA+UVB-induced tyrosinase expression can be observed ().

[0302] 3- Evaluation of melanin content by Fontana-Masson staining on day 7

[0303] When exposed to UVA+UVB treatment, skin models exhibit a marked increase in melanin content, primarily located in the basal layer of the epidermis. Notably, focal areas of high melanin concentration can also be observed in untreated controls ().

[0304] It is interesting to note that the addition of compound C1 to the culture medium of skin models appears to reduce UVA+UVB induced melanin expression ().

[0305] Quantitative analysis confirms a tendency for compound 1 to inhibit UVA+UVB-induced melanin content. However, the differences compared to UVA+UVB-only controls are not statistically significant ().

[0306] In conclusion, the comprehensive data generated in this study suggest that compound C1 is an effective inhibitor of tyrosinase expression in human skin. Therefore, it has the potential to suppress the melanogenesis process, thereby reducing the melanin content present in human skin.

[0307] Example 4: Study of the depigmenting properties of 7 compounds according to the invention

[0308] In this example, the depigmenting effects of two other compounds, compounds 4 and 5, on melanin synthesis in normal human epidermal melanocytes stimulated by L-tyrosine + α-MSH were evaluated. The objective is to assess the efficacy of the compounds in reducing melanin production, which is relevant for treating hyperpigmentation conditions.

[0309] The main objective is to determine the inhibitory effects of the compounds on melanin synthesis under stimulated conditions.

[0310] Melanocytes were cultured in 24-well plates and treated with the tested compounds or a reference compound (lipoic acid) in the presence of L-tyrosine + α-MSH. Melanin content was measured photometrically after 9 days of incubation.

[0311] 2.1 Biological Model

[0312] Cell type: Normal human epidermal melanocytes - Moderately pigmented (NHEM-MP), used in the 8th pass

[0313] Growing conditions: 37°C, 5% CO2

[0314] Culture medium: Dermalife complete medium optimized for testing

[0315] - Test medium: M254 base medium optimized for testing, supplemented with

[0316] HMGS-2 without PMA

[0317] 2.2 Compounds to be tested

[0318] Test CompoundAppearance / StorageStock SolutionTest ConcentrationsRacemic Compound K4Lot No. ELS-942-067MW: 604.9 g / molRF241017-4 Powder Storage: RT1 mM in DMSO 0.1, 0.5 and 2.5 µM (DMSO - 0.25%) Compound K3 Batch No. ELS-942-113 MW: 692.9 g / mol RF241017-3 Powder Storage: RT1 mM in DMSO 0.1, 0.5 and 2.5 µM (DMSO - 0.25%)

[0319] 2.3 Culture and processing

[0320] Melanocytes were seeded in 24-well plates and cultured in culture medium for 24 hours. The medium was then replaced with a test medium containing or not (stimulated control) the tested compounds or the reference compound (lipoic acid tested at 5 µg / mL), and the cells were pre-incubated for 24 hours. After pre-incubation, the medium was replaced with a medium containing or not (control) the tested compounds or the reference compound, as well as the inducer (L-tyrosine + α-MSH tested at 300 µM + 0.1 µM). The cells were then incubated for 9 days, with treatment repeated on days 2 and 6. An unstimulated control was performed concurrently.

[0321] All experimental conditions were carried out in n=3.

[0322] 2.4 Evaluation of melanin content

[0323] At the end of the incubation period, the culture supernatants were removed and the melanin was extracted by cell lysis using a 0.5 N NaOH solution. The optical density (OD) of each experimental point was measured at 405 nm, and the amount of melanin was determined according to a standard melanin curve (standard curve ranging from 0.39 to 100 µg / mL). The results are expressed as µg / mL of melanin.

[0324] 2.5 Data Management

[0325] The raw data was analyzed using Microsoft Excel® software

[0326] Comparisons between groups were performed using an unpaired Student's t-test. The statistical analysis can be interpreted if n ≥ 5; however, for n < 5, the statistical values ​​are given for illustrative purposes only.

[0327] Formula used:

[0328] Standard error of the mean: sem= Sd / n√

[0329] The standard error of the mean (SEM) is a measure of the distance between the sample mean and the true population mean. It is calculated by dividing the standard deviation by the square root of the sample size.

[0330] 3 Results

[0331] At the highest test concentration, all compounds contained 0.25% DMSO. Therefore, a solvent control condition of 0.25% DMSO was performed to identify a potential interfering effect of this organic solvent. As it had no effect, all results were commented on in relation to the stimulated control.

[0332] The results are presented in the table:

[0333] Test Compound Concentration Melanin (µg / ml) Mean (µg / ml) sem (µg / ml) % Stimulated Control sem (%) p (1) Unstimulated Control -10.09.99 9.89.90.1340***Control 29.8-28.728.90.51002-28.3Lipoic Acid 5 µg / ml 12.812.212.70.2441***13.0Compound K 30.1 µM 30.129.430.00.31041ns 30.50.5 µM 26.527.027.60.9953ns29.32.5 µM(DMSO - 0.25%) 8.28.38.20.1280***8.1Compound K 40.1 µM27.626.627.10.3941*27.00.5 µM23.522.622.90.3791***22.62.5 µM(DMSO - 0.25%)10.911.811.60.3401***12.0

[0334] (1): Statistical significance threshold ns: > 0.05, not significant

[0335] * : 0.01 to 0.05, Significant

[0336] ** : 0.001 to 0.01, very significant

[0337] ***: < 0.001, extremely significant

[0338] Treatment of melanocytes with L-tyrosine + α-MSH (300 µM + 0.1 µM) induced a significant stimulation of melanin synthesis (~27.9 µg / ml melanin – mean of the two plates) compared to the unstimulated control condition (~9.5 µg / ml melanin). The reference compound, lipoic acid, tested at 5 µg / ml, inhibited L-tyrosine synthesis.

[0339] + α-MSH (~46% of the stimulated control condition). These results were expected and validated the trial.

[0340] Under the experimental conditions of this test, compound K4, tested at 2.5 µM, induced strong inhibition of melanin synthesis, but this was due more to inhibition of proliferation than to inhibition of melanin production (40% of the stimulated control). When the compound was tested at 0.5 µM, it induced inhibition of melanin synthesis by melanocytes stimulated by L-tyrosine + α-MSH (79% of the stimulated control). Tested at a lower concentration, the compound had no effect.

Claims

Cosmetic and non-therapeutic use of a composition comprising essentially, or consisting of, a compound of the following formula I: (I)or its enantiomers, isomers or diastereomers, or a mixture thereof, or a salt or solvate thereof, where R1 and R2, independently of each other, are a C3 polyol, a C4-C6 sugar, a uronic acid, a glycoside, in particular glucoside, or R1 is OH and R2 is a C4-C6 sugar, or a C3 polyol, or R1 is a C4-C6 sugar, or a C3 polyol, and R2 is OH, provided that if the composition is an extract of Aralia taibaiensis, said composition comprises more than 2.5% by mass relative to the total mass of said composition of said compound, for the decolorization or bleaching of the skin or hair of a healthy individual. Use according to claim 1, wherein said sugar is selected from erythrose, threose, erythrulose, ribose and deoxyribose, arabinose, xylose, lyxose, ribulose, xylulose, allose, altrose, galactose, glucose, gulose, idose, mannose, talose, fructose, psicose, sorbose and tagadose, and deoxyhexoses, namely fucose and rhamnose. Use according to claim 1 or 2, wherein the uronic acids are for example beta-D-Glucuronic acid, alpha-D-Glucuronic acid, alpha-D-glucopyranuronic acid, beta-D-glucopyranuronic acid, alpha-D-galactopyranuronic acid, beta-D-galactopyranuronic acid, beta-L-Glucuronic acid, alpha-L-Glucuronic acid, alpha-L-glucopyranuronic acid, beta-L-glucopyranuronic acid, alpha-L-galactopyranuronic acid, beta-L-galactopyranuronic acid or hexopyranuronic acid. Use according to any one of claims 1 to 3, wherein said compound has the following formula Ia: (Ia)where R1 and R2 are as defined in any one of claims 1 to 3. Use according to any one of claims 1 to 4, wherein the compound is selected from the following compounds: , , , , , , , , , , , , , , , , , , , , , , And . Use according to any one of claims 1 to 5, said compound being Calenduloside F of formula Ib as follows (Ib). Cosmetic composition for topical application comprising from 0.00001 to 1% by mass relative to the total mass of the cosmetic composition of a composition comprising, consisting essentially of, or consisting of a compound as defined in any one of claims 1 to 6. Non-therapeutic cosmetic method for depigmenting or whitening the skin or skin appendages comprising a topical application step of a composition as defined in any one of claims 1 to 6, or of a composition according to claim 7. Formula compound (I), Where R1 and R2 independently of each other are a glycerol, a glucose or a glucuronic acid or R1 is OH and R2 is a C3 polyol or a C4-C6 sugar, provided that the C6 sugar is not glucose, or R1 is a C3 polyol or a C4-C6 sugar provided that the C6 sugar is not glucose, and R2 is OH, and provided that the compound is not the compound of formula: . The compound according to claim 9, said compound being selected from the following compounds: , , , , , , , , , , , , , , , , , , , And .

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