Novel formulations of isothiocyanates, methods for preparation and uses thereof

Abstract

A formulation comprising an isothiocyanate (ITC) with a sulfoxide moiety and a combination of at least two of the following compounds: - organic acids, - amino acids, - ectoine and 5-hydroxyectoine, - compounds comprising a tetramethyl ammonium moiety, - phenols, - glycols, - glycerol, - sugar alcohols, - simple saccharides, - terpenoids, - chaotropic agents, wherein the above-mentioned compounds may be mixed in any combination with the selected isothiocyanate. Products comprising said formulation. A method of preparing said formulation. A method of stabilizing isothiocyanate in said formulation.

Description

[0001]NOVEL FORMULATIONS OF ISOTHIOCYANATES, METHODS FOR PREPARATION AND USES THEREOF Field of the invention The present invention belongs to the field of compositions comprising isothiocyanates. The invention relates to novel formulations of isothiocyanates, particularly sulforaphane and sulforaphene, methods for preparation and uses thereof. Background of the invention and the technical problem Isothiocyanates are naturally occurring small molecules that are formed from glucosinolate precursors of cruciferous vegetables. Many isothiocyanates, both natural and synthetic, display anticarcinogenic activity because they reduce activation of carcinogens and increase their detoxification. Two such examples are sulforaphane (SFN) and sulforaphene (SFEN). Sulforaphane or 1-Isothiocyanato-4-(methanesulfinyl)butane (SFN) with structural formula is a compound formed upon damage to cruciferous vegetables such as chewing or chopping, as the enzyme myrosinase transforms glucoraphanin into SFN. Typical sources are broccoli, Brussels sprouts, cauliflower sprouts, kale, colards, arugula, kohlrabi, mustard, turnip, watercress, and cabbage. It can also be produced in the gut by certain bacteria (Conaway et al., Nutr Cancer 2000;38(2):168-78). SFN has two possible stereoisomers due to the presence of a stereogenic sulfur atom. The R-sulforaphane enantiomer occurs naturally, while the S- sulforaphane can be synthesized, which is also well known to the person skilled in the art. SFN exhibits antioxidant (Reidl et al., Immunol 2009;130:244-251), antimicrobial (Fahey et al., PNAS 2002;99:7610-7615; Johansson et al., Planta Med 2008;74:747-750), as well as anti- inflammatory properties (Heiss et al., J Biol Chem 2001;276:32008-32015; Ritz et al., Am J Physiol Lung Cell Mol Physiol 2007;292:L33-L39). The potential health effects on various conditions, including cancer prevention, blood sugar, heart disease, paved the way to several clinical trials to further assess its effects on various cancers, cardiovascular disease, upper airway inflammation, radiation dermatitis, and vascular health (entries available at www.clinicaltrials.gov). Sulforaphene (IUPAC (E)-4-isothiocyanato-1-methylsulfinylbut-1-ene) (SFEN) with formula is a natural product found in for example Thulinella chrysantha, Matthiola incana, and Raphanus sativus with data available about its role in apoptosis, protective function against reactive oxygen species, radical scavenging activity as well as anti-tumour activity. Due to all possible uses of isothiocyanates, particularly SFN and SFEN a need for novel formulations exists, particularly if combined with other possible active ingredients. Prior art Several different combinations with SFN are known in the prior art. In FR2888235 SFN was combined with arabic gum, acacia gum to encapsulate it. Similarly, patent application EP2726087 describes SFN encapsulated by a compound selected from the group comprising acacia gum, maltodextrin and the mixture thereof. Garcia-Saldana et al. (2016; https: / / doi.org / 10.1016 / j.foodchem.2016.01.087) disclose combinations of SFN with gelatin / gum arabic and gelatin / pectin. Tian et al. (2015: https: / / doi.org / 10.1016 / j.carbpol.2015.01.003) combined chitosan, acetic acid and SFN. US2014170218A discloses an extraction method that provides a non-aqueous extract from Brassica Oleracea Italica, with low content of erucic acid and lipids, containing at least 30% by weight of SFN. This non-aqueous extract is stabilized by means of encapsulation into a polysaccharide matrix selected from acacia gum, maltodextrin and mixture thereof, leading to a powder containing at least 10% by weight of SFN. No further data are provided with respect to the stabilization effect. The suggested solution for stabilizing SFN is limiting, since the produced encapsulated composition is only used as such in preparing further nutraceutical compositions. Furthermore, it is not apparent if and how higher concentrations of SFN can be reached by this method or if it is applicable in SFN of synthetic origin. EP2120969A1combines cyclodextrins with SFN. The theoretical loading of the SFN- cyclodextrin complexes prepared therein ranges from 0.94 to 13.3 %. Similarly, WO2008091608A1, EP2796140, Recio et al. (2017; https: / / doi.org / 10.1016 / j.carbpol.2017.12.022), Wu et al. (2010; doi:10.1016 / j.carbpol.2010.05.020) combine SFN with at least one cyclodextrin to form a complex between the SFN and the at least one cyclodextrin. Patent application CN102423492A proposes a specific combination of SFN and propylene group-beta- cyclodextrin in ratio 1:0.2～4 for preparation of a cancer therapy drug. US9254331B2 discloses galenic compositions comprising SFN in an amount ranging from 0.01 to 15% and at least one anhydrous ester, the main chain and / or optionally the branched chains of which are free from and / or reactive groups or functions in an amount ranging from 0.1 to 99.9% by weight, based on the total weight of the cosmetically and pharmaceutically acceptable composition. US2011003747A describes compositions prepared for topical treatment (e.g. example 3) that comprise SFN and jojoba oil. EP2163238A2 discloses combination of SFN with dicarboxylic acids for the preparation of cosmetic or dermatological composition for lightening the skin. The cosmetic or dermatological compositions comprise further ingredients. US6511675B2 discloses a dietary supplement composition comprising SFN, naringen, hesperidin, narirutin, quercetin, β-carotene, lutein, lycopene, and isoflavones. Patent application EP4138579 discloses a combination of SFN with fatty acids, which exhibit a stabilizing effect in comparison to neat SFN. Ko et al. (2013; https: / / doi.org / 10.1016 / j.biomaterials.2013.03.066) disclose a combination of Poly(D,L-lactic-co-glycolic) acid (PLGA) and SFN, while patent CN103877066B discloses a combination of hyaluronic acid decorated PLGA, docetaxel and SFN. Danafar et al. (2016: http: / / dx.doi.org / 10.3109 / 10837450.2016.1146296) prepared a combination of 20 mg mPEG–PCL copolymer and 6 mg SFN. Several authors combined bovine serum albumin and SFN, such as Do et al., (2010: doi:10.1016 / j.ijpharm.2009.11.009), Dong et al. (2014; https: / / doi.org / 10.1016 / j.jlumin.2013.09.078) and Wu (2014; https: / / doi.org / 10.1016 / j.carbpol.2013.11.057). US9254331 discloses a combination of an anhydrous ester and SFN. Wang et al (2011: doi:10.4028 / www.scientific.net / MSF.687.539) prepared a mixture of chitosan, alginate and SFN. Description of the solution to the technical problem The above-mentioned prior art documents combined SFN with various polymers (usually polysaccharides) or complex organic acids. It would be, however, desirable to provide novel formulations of isothiocyanates (ITCs) comprising a sulfoxide moiety, such as sulforaphane (SFN) and sulforaphene (SFEN) with compounds or compositions that would allow use in various fields, such as pharmaceutical industry, food industry, cosmetic industry, etc. It is an additional aim of the present invention to provide formulations which comprise SFN in higher concentration. It is a further object of the present invention to provide a method for preparation of novel formulations. The technical problem is solved as defined in the independent claims, wherein preferred embodiments of the invention are defined in the dependent claims. For better understanding of the invention as well as the description the following definitions and abbreviations are hereby provided. Definitions a dietary supplement a composition consumed in order to supplement the diet for a nutritional purpose. Dietary supplements may take a variety of forms including but not limited to pills, capsules, tablets and liquid forms. a food additive a composition intended to be added to a food product in order to enhance the nutritional value of a diet that includes said food product. nutraceutical a composition based on a food or foods possessing a positive composition or functional and significant effect on health and on the prevention of food pathologies pharmaceutical a physical mixture containing a therapeutic compound to be composition administered to a mammal, e.g., a human in order to prevent, treat or control a particular disease or condition affecting the mammal cosmetic composition compositions which are used to treat, care for or improve the appearance of the skin and / or the scalp eutectic mixture physical mixture of two or more components that solidifies at a single temperature lower than the crystallization point of any individual component Eutectic-like mixture a liquid mixture / system / solvent comprising two or more components that exhibits characteristics similar to a eutectic mixture / system / solvent, enabling it to remain in a liquid state at a specific desired temperature deep eutectic a eutectic mixture whose components present enthalpic-driven mixture / system / solvent negative deviations from thermodynamic ideality ratio molar ratio, unless indicated otherwise in the text Abbreviations Bet betaine C10 decanoic acid C8 octanoic acid C18:2 linoleic acid CA citric acid Cam camphor DES deep eutectic solvent Ect ectoine EG ethylene glycol Fru fructose Glc glucose Gly glycerol ITC isothiocyanate LA lactic acid MA malic acid Me menthol PG propylene glycol Phe phenylalanine Pro proline SFN sulforaphane SFEN sulforaphene Suc sucrose U urea TA tartaric acid Treh trehalose Ty thymol The essence of the invention is that the ITC, preferably SFN or SFEN, is mixed with a combination of at least two, optionally three or more, of the following compounds: − organic acids, preferably selected in the group comprising decanoic acid, octanoic acid, linoleic acid, citric acid, lactic acid, malic acid, tartaric acid, oleic, other omega3 acids (such as (EPA), (DHA), a-linoleic (ALA)), omega6 acids (such as arachidonic (ARA), gamma- linoleic (GLA), dihomo-gamma-linoleic (DGLA)) acid, − amino acids, preferably selected in the group comprising proline and phenylalanine, − ectoine and 5-hydroxyectoine, − compounds comprising a tetramethyl ammonium moeity, preferably selected in the group comprising betaine, sarcosine, choline, carnitine, trimethylamine N-oxide (TMAO), glycerophosphorylcholine (GPC) and phosphatidylcholine (PC) and their salts, − phenols, − glycols, i.e. compounds having two hydroxyl (―OH) groups attached to different carbon atoms, preferably ethylene glycol, propylene glycol, butylene glycol, polyethylene glycol (PEG), − glycerol, − sugar alcohols, such as sorbitol, xylitol and similar compounds, − simple saccharides, preferably selected in the group comprising glucose, fructose, sorbose, sucrose, trehalose, − terpenoids, such as camphor, menthol, and thymol, − chaotropic agents, preferably selected in the group comprising urea, thiourea, guanidine, arginine and their salts, wherein the above-mentioned compounds may be mixed in any combination with the selected isothiocyanate. The combination of compounds to be mixed with the ITC may form a eutectic or a eutectic-like mixture. In some embodiments, the combination of compounds to be mixed with the ITC forms a deep eutectic mixture (DES). In the formulations according to the invention, the isothiocyanate is of natural, synthetic, biosynthetic or semi-synthetic origin. According to preferred embodiments the selected ITC is SFN or SFEN. However, the invention covers ITCs with different carbon chain lengths. These formulations represent novel combinations of ITCs with natural, renewable, sustainable compounds, which also represent different active ingredients that show suitability for use in various products, such as pharmaceutical, nutritional or cosmetic products. The ITC concentration in the formulations according to the invention is any suitable concentration, preferably up to 60% (w / w), most preferably between 20 and 60% (w / w). The combinations of components other than ITC may be prepared in any molar ratios, wherein the molar ratio of each component in the combination (excluding the ITC) with respect to the total moles of the combination (excluding the ITC) may range from 0.01 to 0.9. The preferred combinations of ITC with one or more of the above-mentioned compounds are: − ITC : Me : C18:2, − ITC : Me : C10 − ITC : Ty: C18:2, − ITC : Ty: C8, − ITC : Ty: C10, − ITC : Pro: C8, − ITC : Pro : C18:2 : C8, − ITC : Cam : PA, − ITC : Me : Cam, − ITC : Ty : PG : C18:2, − ITC : Bet : PG, − ITC : Ect : PG, − ITC : PG : C18:2, − ITC : Bet : Ect : PG : C18:2, − ITC : Glc : CA, − ITC : LA : Gly, − ITC : LA : Pro, − ITC : Bet : LA, − ITC : Bet : LA : Gly, − ITC : Bet : LA : U, − ITC: Ect : LA, − ITC: Ect : LA : Gly, − ITC : MA : Gly, − ITC : CA : Gly, − ITC : TA : Gly, − ITC : Bet : Ect : LA : Gly, − ITC : Bet : Ect : MA : Gly, − ITC : Bet : Ect : CA : Gly, − ITC : Bet : Ect : TA : Gly, − ITC : Bet : Suc : Pro : LA : Gly, − ITC : Bet : Suc : Phe : MA : Gly, − ITC : Bet : Suc : Ect : CA : Gly, − ITC : Bet : Treh : Phe : MA : Gly, and − ITC : Bet : Treh : Ect : LA : Gly. The ITC concentration in the above-given examples is any suitable concentration, preferably up to 60% (w / w), most preferably between 20 and 60% (w / w). The combinations of components are in any molar ratio, wherein the molar ratio of each component (for example Me or C18:2) of the combination (for example Me and C18:2) with respect to the total moles of the combination may range from 0.01 to 0.9. The preferred combinations of SFN with one or more of the above-mentioned compounds are: − SFN : Me : C18:2, − SFN : Me : C10, − SFN : Ty: C18:2, − SFN : Ty: C8, − SFN : Ty: C10, − SFN : Pro: C8, − SFN : Pro : C18:2 : C8, − SFN : Cam : PA, − SFN : Me : Cam, − SFN : Ty : PG : C18:2, − SFN : Bet:PG, − SFN : Ect : PG, − SFN : PG : C18:2, − SFN : Bet : Ect : PG : C18:2, − SFN : Glc : CA, − SFN : LA : Gly, − SFN : LA : Pro, − SFN : Bet : LA, − SFN : Bet : LA : Gly, − SFN : Bet : LA : U, − SFN : Ect: LA, − SFN : Ect : LA : Gly, − SFN : MA : Gly, − SFN : CA : Gly, − SFN : TA : Gly, − SFN : Bet : Ect : LA : Gly, − SFN : Bet : Ect : MA : Gly, − SFN : Bet : Ect : CA : Gly, − SFN : Bet : Ect : TA : Gly, − SFN : Bet : Suc : Pro : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, − SFN : Bet : Suc : Ect : CA : Gly, − SFN : Bet : Treh : Phe : MA : Gly, and − SFN : Bet : Treh : Ect : LA : Gly. The formulations as described above may comprise other components, which may be selected in the group disclosed above for the preparation of the formulations or any other components, not used in the preparation of formulations described above. The inventors have furthermore surprisingly found that some formulations according to the invention stabilize SFN of natural or synthetic origin, such that storage at temperatures higher than 0 °C is facilitated. The prepared formulations with SFN showed stabilization of SFN under 4°C and / or bench conditions (20-25°C) over a 28-day period. A composition is defined as »stabilized«, when the isothiocyanate in said composition degrades less or more slowly than it does on its own, under the same conditions. These formulations are: − SFN : Me : C18:2, − SFN : Me : C10, − SFN : Ty : C18:2, − SFN : Ty : C8, − SFN : Ty : C10, − SFN : Me : Cam, − SFN : Ty : PG:C18:2, − SFN : Ect : PG, − SFN : Bet : Ect : PG: C18:2, − SFN : Glc : CA, − SFN : LA : Gly, − SFN : Bet : LA : Gly, − SFN : MA : Gly, − SFN : CA : Gly, − SFN : TA : Gly, − SFN : Bet : Ect : TA : Gly, − SFN : Bet : Suc : Pro : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, − SFN : Bet : Suc : Ect : CA : Gly, − SFN : Bet : Treh : Phe : MA : Gly, and − SFN : Bet : Treh : Ect : LA : Gly. In preferred embodiments, hydrophobic combinations of compounds to be mixed with the isothiocyanate, are anhydrous, whereas hydrophilic ones may be prepared with a minimal amount of water to achieve a liquid state at room temperature. Even more surprisingly, some embodiments of the formulations showed improved long-term stability monitored for 60 and 90 days, respectively. Said embodiments are for example: − SFN : Me : C18:2, − SFN : Me : C10, − SFN : Ty : C18:2, − SFN : LA : Gly − SFN : MA : Gly, − SFN : CA : Gly, − SFN: TA : Gly, − SFN : Bet : Suc : Pro : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, − SFN : Bet : Suc : Ect : CA : Gly, − SFN : Bet : Treh : Phe : MA : Gly, and − SFN : Bet : Treh : Ect : LA : Gly. Furthermore, some formulations allow increased SFN concentrations, such 40% (w / w) or 60% (w / w). Some examples of formulations showed improved stability at these higher loadings as well, which is particularly useful for the preparation of pharmaceutical products. These formulations with SFN at concentrations up to 60% (w / w) are selected in the group comprising: − SFN : LA : Gly, − SFN : Ty : C18:2, − SFN : MA : Gly, − SFN : TA : Gly, − SFN : CA : Gly, − SFN : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, − SFN : Bet : Suc : Ect : CA : Gly. Stability was observed at both 4 °C and 28 °C, wherein stability at 4 °C was higher. The formulations as described above may comprise other components, which may be selected in the group disclosed above for the preparation of the formulations or any other components, not used in the preparation of formulations described above. Among above-described formulations are composed solely of components deemed safe for human use. The majority is comprising bioactive compounds preferable for topical or oral use (such as betaine, ectoine and linoleic acid), as the priority was given to components that offer benefits for dietary or other health-related purposes, such as linoleic acid, betaine and ectoine, trehalose and essential amino acids. In one aspect of the invention a method of preparing a formulation comprising an isothiocyanate is provided. The method comprises the steps of: i. Preparing a combination of at least two components selected in the group consisting of: o organic acids, preferably selected in the group comprising decanoic acid, octanoic acid, linoleic acid, citric acid, lactic acid, malic acid, tartaric acid, oleic, other omega3 acids (such as (EPA), (DHA), a-linoleic (ALA)), omega6 acids (such as arachidonic (ARA), gamma-linoleic (GLA), dihomo-gamma-linoleic (DGLA)) acid, o amino acids, preferably selected in the group comprising proline and phenylalanine, o ectoine and 5-hydroxyectoine, o compounds comprising a tetramethyl ammonium moiety, preferably selected in the group comprising betaine, sarcosine, choline, carnitine, trimethylamine N-oxide (TMAO), glycerophosphorylcholine (GPC) and phosphatidylcholine (PC) and their salts, o phenols, o glycols, i.e. compounds having two hydroxyl (―OH) groups attached to different carbon atoms, preferably ethylene glycol, propylene glycol, butylene glycol, polyethylene glycol (PEG), o glycerol, o sugar alcohols, such as sorbitol, xylitol and similar compounds, o simple saccharides, preferably selected in the group comprising glucose, fructose, sorbose, sucrose, trehalose, o terpenoids, such as camphor, menthol, and thymol, o chaotropic agents, preferably selected in the group comprising urea, thiourea, guanidine, arginine and their salts, ii. Mixing the prepared combination and the selected isothiocyanate, preferably sulforaphane or sulforaphene at the desired concentration, and iii. Optionally adding any further components. It is further possible to perform a step of water removal, either before or after step ii. In one aspect of the invention a method of stabilizing an isothiocyanate is provided. The method comprises the steps of: a) Preparing a combination of at least two components selected in the group consisting of: o organic acids, preferably selected in the group comprising decanoic acid, octanoic acid, linoleic acid, citric acid, lactic acid, malic acid, tartaric acid, oleic, other omega3 acids (such as (EPA), (DHA), a-linoleic (ALA)), omega6 acids (such as arachidonic (ARA), gamma-linoleic (GLA), dihomo-gamma-linoleic (DGLA)) acid, o amino acids, preferably selected in the group comprising proline and phenylalanine, o ectoine and 5-hydroxyectoine, o compounds comprising a tetramethyl ammonium moiety, preferably selected in the group comprising betaine, sarcosine, choline, carnitine, trimethylamine N-oxide (TMAO), glycerophosphorylcholine (GPC) and phosphatidylcholine (PC) and their salts, o phenols, o glycols, i.e. compounds having two hydroxyl (―OH) groups attached to different carbon atoms, preferably ethylene glycol, propylene glycol, butylene glycol, polyethylene glycol (PEG), o glycerol, o sugar alcohols, such as sorbitol, xylitol and similar compounds, o simple saccharides, preferably selected in the group comprising glucose, fructose, sorbose, sucrose, trehalose, o terpenoids, such as camphor, menthol, and thymol, o chaotropic agents, preferably selected in the group comprising urea, thiourea, guanidine, arginine and their salts, b) Mixing the prepared combination and the selected isothiocyanate, preferably sulforaphane or sulforaphene at the desired concentration, c) Optionally adding further components, and d) Storing at any temperature, preferably higher than 0 °C. Advantageously, the compositions disclosed in the present invention may be used either alone or as a component in the compositions of pharmaceutical, food or drink products, nutraceutical products, dietary supplements, food additives, cosmetic, skin or hair products, feed additives, and agricultural products. Detailed description of the invention The invention will be described in further detail based on exemplary embodiments and examples. The exemplary embodiments of the formulations according to the invention, wherein the concentration of SFN is arbitrary, preferably from 20 to 60% (w / w), are: − SFN : Me : C18:2, wherein the molar ratio of Me and C18:2 is preferably 1:1, 1:3 or 1:5, − SFN : Me : C10, − SFN : Ty: C18:2, wherein the molar ratio of Ty and C18:2 is preferably 1:3, 1:5, or 1:7, − SFN : Ty: C8, − SFN : Ty: C10, − SFN : Pro: C8, − SFN : Pro : C18:2 : C8, − SFN : Cam : PA, − SFN : Me : Cam, − SFN : Ty : PG : C18:2, − SFN : Bet:PG, − SFN : Ect : PG, − SFN : PG : C18:2, − SFN : Bet : Ect : PG : C18:2, wherein the molar ratio of Bet:Ect:PG:C18:2 is preferably 1:1:15:6 or 1:1:15:15, − SFN : Glc : CA, − SFN : LA : Gly, wherein the molar ratio of LA and Gly is preferably 1:1 or 1:2, − SFN : LA : Pro, − SFN : Bet : LA, − SFN : Bet : LA : Gly, − SFN : Bet : LA : U, − SFN : Ect: LA, − SFN : Ect : LA : Gly, − SFN : MA : Gly, wherein the molar ratio of MA and Gly is preferably 1:2.5 or 1:3, − SFN : CA : Gly, wherein the molar ratio of CA and Gly is preferably 1:1; 1:2.5 or 1:3, − SFN : TA : Gly, wherein the molar ratio of TA and Gly is preferably 1:2.5 or 1:3, − SFN : Bet : Ect : LA : Gly, − SFN : Bet : Ect : MA : Gly, − SFN : Bet : Ect : CA : Gly, − SFN : Bet : Ect : TA : Gly, − SFN : Bet : Suc : Pro : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, − SFN : Bet : Suc : Ect : CA : Gly, − SFN : Bet : Treh : Phe : MA : Gly, and − SFN : Bet : Treh : Ect : LA : Gly. The possible water content in the formulations is given in the table below: Formulation with SFN molar ratio water content (additional components) (w / w) Me : C18:2 1:1 - Me : C18:2 1:3 - Me : C18:2 1:5 - Me : C8 1:1 - Ty : C18:2 1:1 - Ty : C18:2 1:3 - s Ty : C18:2 1:5 - noitTy : C8 1:3 - aluTy : C10 1:1 - mrof Pro : C8 1:4 - ciboPro: C18:2:C8 1:4:2 - hporCam : PA 1:1 - dyMe : Cam 1:1 - HdesTy : PG : C18:2 1:1:1 -ab-lBet : PG 1:4 -ocylEct : PG 1:12 -gPG : C18:2 1:1 - se nEct : PG : C18:2 1:12:6 -n oiel ty alBet : Ect : PG: C18:2 1:1:15:6 -puor mrBet : Ect : PG: C18:2 1:1:15:15PofGlc : CA 1:1:6.5(H2O)24 LA : Pro 2:1:1(H2O)6 Bet : LA : U 1:2:0.5:1(H2O)7 Ect : LA 1:2:1(H2O)7 Glc : LA 1:2:1(H2O)5snLA : Gly 1:1:1o(H2O)6italLA : Gly 1:2:1 5u (H2O)mrBet : LA 1:4:0.5 5o(H2O)ftnBet : LA : Gly 1:1:2:0.5 3e(H2O)noEct : LA : Gly 1:3:2:2p (H2O)6 moBet : Ect : LA : Gly 0.5:0.5:3:2:2(H2O)6c-eeMA : Gly 1:2.5:2(H2O)9rht MA : Gly 1:3 0dnaCA : Gly 1:1:1.5 -(H2O)9owtCA : Gly 1:2.5:2(H2O)8cilliCA : Gly 1:3 0hporTA : Gly 1:2.5:2(H2O)11dyTA : Gly 1:3 0 HtneEct : Bet : MA : Gly 0.5:0.5:1:2:2(H2O)7,5nopEct : Bet : CA : Gly 0.5:0.5:1:2:2(H2O)7mocEct : Bet : TA : Gly 0.5:0.5:1:3:3 i(H2O)7tluBet : Suc : Pro : LA : Gly 0.4:0.1:0.1:2:0.5:3(H215 mO)Bet : Suc : Phe : MA : Gly* 0.4:0.1:0.1:2:0.6:4(H2O)15 Bet : Suc : Phe : MA : Gly** 0.4:0.1:0.1:2:5 0cil sli noiBet : Treh : Phe : MA : Gly 0.4:0.1:0.1:2:0.6:4(H2O)14hptoarluBet : Tre : Ect : LA : Gly 0.4:0.1:0.1:2:0.6:1(H2O)5dymrBet : Suc : Ect : CA : Gly 0.25:0.1:0.1:1:1:2.5 11 Hof (H2O) *components were mixed in a suitable molar ratio, followed by freeze-drying of the mixture before the addition of SFN ** hydrophillic DES prepared without water The method of preparing the above-mentioned formulations comprises the steps of: i. Preparing a combination of at least two components for mixing with the SFN as described above, and ii. Mixing the prepared combination and the SFN at the desired concentration. It is further possible to perform a step of water removal, either before step ii. or after it. In some embodiments the combination of at least two components in step i. results in formulation of a eutectic or eutectic-like mixture, in some cases a DES. The mixtures were prepared as follows: the appropriate amount of the components was placed in a 10 mL flask and the mixture was heated at 60 °C with stirring for 1 or 2 hours until a clear homogeneous liquid was formed. Upon cooling to room temperature, the mixture was left on a bench for a few days in order to observe possible solidification or precipitation. Before use, the mixture forming compounds were dried under vacuum. In some embodiments, prepared mixtures were prepared as described and then freeze-dried. Some of the above-mentioned formulations mentioned above stabilize SFN at temperatures higher than 0 °C, which was tested at 4°C and bench conditions (room temperature, 20-25°C) over a 28-day period. A composition is defined as "stabilized", when the isothiocyanate in said composition degrades less or more slowly than it does on its own, under the same conditions. The general procedure for SFN stability screening is as follows: in a dark vial the appropriate amount of SFN was added to the respective amount of the eutectic medium, to obtain a total mass 240 mg of the mixture, which was thoroughly homogenised, capped, and stored at selected temperatures. At specific intervals of time (up to three months), 5 µL of mixture was withdrawn, diluted with dimethyl sulfoxide or acetonitrile (V = 995 µL) and analysed by HPLC according to the method described in WO 2021 / 214188 A1. The stability of SFN was expressed as a ratio of the initial SFN concentration and the concentration after incubation under certain conditions (SFN retention, %). Neat SFN was used as control, and it was noted that when neat SFN was stored over a 28-day period at 4°C or under bench conditions (incubation at temperature from 20 to 25 °C), its concentration decreased to 75% or 51% of the initial concentration, respectively. A positive control was used according to EP4138579 disclosing stabilization of SFN in fatty acids, such as C18:2. This control was used to evaluate the level of SFN stabilization. The amount of SFN in the formulation was 20% (w / w). These preferred embodiments of the formulations and the SFN retention after 28 days are: Formulation or control SFN retention (%) after 28 days at 4 °C SFN (control) 75.2 SFN : C18:2 (positive control) 101.5 SFN : Me : C18:2 in ratio Me : C18:21:1 99.9 SFN : Me : C18:2 in ratio Me : C18:21:3 85.5 SFN : Me : C18:2 in ratio Me : C18:21:5 83.6 SFN : Me : C10 in ratio Me : C101:1 79.9 SFN : Ty: C18:2 in ratio Ty : C18:21:3 101.8 SFN : Ty: C18:2 in ratio Ty : C18:21:5 88.5 SFN : Ty: C18:2 in ratio Ty : C18:21:7 102.1 SFN : Ty: C8 in ratio Ty : C8 1:3 82.3 SFN : Ty: C10 in ratio Ty : C101:1 79.3 SFN : Me : Cam in ratio Me : Cam 1:1 59.4 SFN : Ty : PG : C18:2 in ratio Ty : PG : C18:21:1:1 74.1 SFN : Ect : PG in ratio Ect : PG 1:12 74.1 SFN:Bet:Ect: PG: C18:2 in ratio Bet:Ect:PG:C18:2 81.7 1:1:15:15 SFN : LA : Gly in ratio LA : Gly 1:1 (freeze dried*) 105.0 SFN : LA : Gly in ratio LA : Gly 1:2 97.4 SFN : MA : Gly in ratio MA : Gly 1:2.5 98.0 SFN : MA : Gly in ratio MA : Gly 1:3 105.2 SFN : CA : Gly in ratio CA : Gly 1:1 100.0 SFN : CA : Gly in ratio CA : Gly 1:2.5 103.8 SFN : CA : Gly in ratio CA : Gly 1:3 98.2 SFN : TA : Gly in ratio TA : Gly 1:2.5 102.2 SFN : TA : Gly in ratio TA : Gly 1:3 102.3 SFN : Bet : Ect : TA : Gly 86.8 in ratio Bec : Ect : TA : Gly 0.5:0.5:1:3 SFN : Bet : Suc : Pro : LA : Gly 103.6 in ratio Bet : Suc : Pro : LA : Gly 0.4:0.1:0.1:2:0.5 SFN : Bet : Suc : Phe : MA : Gly 105.5 in ratio Bet : Suc : Phe : MA : Gly 0.4:0.1:0.1:2:0.6) SFN : Bet : Suc : Ect : MA : Gly 105.2 in ratio Bec : Suc : Ect : MA : Gly 0.4:0.1:0.1:2:5 SFN : Bet : Suc : Ect : CA : Gly in ratio Bet : Suc : Ect : 86.4 CA : Gly 0.25:0.1:0.1:1:1 SFN : Bet : Treh : Phe : MA : Gly 100.9 in ratio Bet : Treh : Phe : MA : Gly 0.4:0.1:0.1:2:0.6 SFN : Bet : Treh : Ect : LA : Gly 95.2 in ratio Bet : Treh : Ect : LA : Gly 0.4:0.1:0.1:2:0.6 *LA (in a water solution containing 90% LA, w / w) and Gly were mixed in a 1:1 molar ratio, followed by freeze-drying of the mixture before the addition of SFN) Formulation or control SFN retention (%) after 28 days at room temperature SFN (control) 51.2 SFN : C18:2 (positive control) 93.7 SFN : Me : C18:2 in ratio Me:C18:21:1 87.9 SFN : Me : C18:2 in ratio Me:C18:21:3 73.6 SFN : Me : C18:2 in ratio Me:C18:21:5 71.0 SFN : Me : C10 in ratio Me:C101:1 72.1 SFN : Ty: C18:2 in ratio Ty:C18:21:3 101.2 SFN : Ty: C18:2 in ratio Ty:C18:21:5 89.1 SFN : Ty: C18:2 in ratio Ty:C18:21:7 97.6 SFN : Ty: C8 in ratio Ty:C81:3 75.8 SFN : Ty: C10 in ratio Ty:C101:1 85.5 SFN : Me :Cam in ratio Me : Cam 1:1 53.2 SFN : Ty : PG : C18:2 in ratio Ty : PG : Lin A 1:1:1 64.2 SFN : Ect : PG in ratio Ect : PG 1:12 68.2 SFN : LA : Gly in ratio LA : Gly 1:1 (freeze dried*) 100.1 SFN : LA : Gly in ratio LA : Gly 1:2 83.8 SFN : MA : Gly in ratio MA : Gly 1:2.5 53.2 SFN : CA : Gly in ratio CA : Gly 1:2.5 85.3 SFN : CA : Gly in ratio CA : Gly 1:3 102.6 SFN: TA : Gly in ratio TA : Gly 1:2.5 74.0 SFN: TA : Gly in ratio TA : Gly 1:3 85.1 *LA (in a water solution containing 90% LA, w / w) and Gly were mixed in a 1:1 molar ratio, followed by freeze-drying of the mixture before the addition of SFN). Even more surprisingly, some embodiments of the formulations showed improved long-term stability monitored for 60 and 90 days, respectively. Stability testing was performed in the same manner as described above; the only difference was the length of incubation. Said embodiments are: − SFN : Me : C18:2, wherein the molar ratio of Me and C18:2 is preferably 1:1, 1:3 or 1:5, − SFN : Me : C10, − SFN : Ty : C18:2, wherein the molar ratio of Ty and C18:2 is preferably 1:3, 1:5 or 1:7, − SFN : LA : Gly, wherein the molar ratio of LA and Gly is preferably 1:1 or 1:2 − SFN : MA : Gly, wherein the molar ratio of MA and Gly is preferably 1:2.5 or 1:3, − SFN : CA : Gly, wherein the molar ratio of CA and Gly is preferably 1:1; 1:2.5 or 1:3, − SFN : TA : Gly, wherein the molar ratio of TA and Gly is preferably 1:2.5 or 1:3, − SFN : Bet : Suc : Pro : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, − SFN : Bet : Suc : Ect : CA : Gly, − SFN : Bet : Treh : Phe : MA : Gly, and − SFN : Bet : Treh : Ect : LA : Gly. Results for this example are shown in the table below for 4 °C, wherein incubation was 60 and 90 days: Formulation or control SFN retention (%) SFN retention (%) after 60 days at 4 °C after 90 days at 4 °C SFN (control) 68.2 65.1 SFN : Me : C18:2 in ratio Me : C18:21:1 98.00 89.7 SFN : Ty: C18:2 in ratio Ty : C18:21:3 104.0 103.6 SFN : Ty: C18:2 in ratio Ty : C18:21:7 104.8 103.2 SFN : LA : Gly in ratio LA : Gly 1:1 98.2 / SFN : LA : Gly in ratio LA : Gly 1:1 (freeze 105.1 102.3 dried*) SFN : LA : Gly in ratio LA : Gly 1:2 101.2 / SFN : MA : Gly in ratio MA : Gly 1:3 85.3 69.1 SFN : CA : Gly in ratio CA : Gly 1:1 105.2 103.5 SFN : CA : Gly in ratio CA : Gly 1:2.5 104.3 105.5 SFN : CA : Gly in ratio CA : Gly 1:3 101.3 105.3 SFN : TA : Gly in ratio TA : Gly 1:2.5 105.3 99.1 SFN : TA : Gly in ratio TA : Gly 1:3 102.4 103.3 SFN : Bet : Suc : Phe : MA : Gly 99.3 / in ratio Bet : Suc : Phe : MA : Gly 0.4:0.1:0.1:2:0.6 SFN : Bet : Suc : Ect : MA : Gly 94.1 90.5 in ratio Bet : Suc : Ect : MA : Gly 0.4:0.1:0.1:2:5 SFN : Bet : Suc : Ect : CA : Gly in ratio Bet 90.3 78.2 : Suc : Ect : CA : Gly 0.25:0.1:0.1:1:1 SFN : Bet : Treh : Phe : MA : Gly 75.5 / in ratio Bet : Treh : Phe : MA : Gly 0.4:0.1:0.1:2:0.6 *LA (in a water solution containing 90% LA, w / w) and Gly were mixed in a 1:1 molar ratio, followed by freeze-drying of the mixture before the addition of SFN). / : measurement not performed or not possible These formulations were particularly stable at 4 °C, whereas best long-term stability at room temperature was observed for formulations SFN : Me : C18:2 in any ratio (retention at 60 days was 65.9%), preferably in ratio of Me and C18:21:1, 1:3 or 1:5, and SFN : Ty: C18:2 in any ratio, preferably in ratio of Ty and C18:21:1, or 1:7 (retention at 60 days was 89.2 and 87.2, respectively). At room temperature the neat SFN had a retention % at 60 days and 90 days, respectively, 45.5 and 42.2%. The stable SFN formulations are suitable for long term storage without complex techniques, such as encapsulation or complexation. The compositions provided by this invention stabilize SFN at temperatures higher than the temperature required to preserve neat SFN (-15°C). Furthermore, some formulations allow increased SFN concentrations, such 40% (w / w) or 60% (w / w), which is particularly useful for the preparation of pharmaceutical products. Stability testing at higher SFN loadings was performed in the same manner as described above, wherein the incubation period was 28 days. These formulations with SFN at concentrations up to 60% (w / w) are selected in the group comprising: − SFN : LA : Gly, − SFN : Ty: C18:2, − SFN : MA : Gly, − SFN : TA : Gly, − SFN : CA :Gly, − SFN : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, − SFN : Bet : Suc : Ect : CA : Gly, Stability was observed at both 4 °C and 28 °C, wherein stability at 4 °C was higher. Some formulations were not tested (indicated by NA), while some formulations showed immeasurable SFN amounts (indicated by / ). The results are shown in the tables below: SFN retention (%) after 28 days at 4 °C Formulation SFN loading 40% 60% SFN : LA : Gly in ratio LA : Gly 1:1 (60% freeze dried*) 94.9 100.2 SFN : Ty : C18:2 in ratio Ty : C18:21:1 74.9 61.6 SFN : MA : Gly in ratio MA : Gly 1:3 NA 90.8 SFN : TA : Gly in ratio TA : Gly 1:3 NA 101.2 SFN : CA : Gly in ratio CA : Gly 1:1 89.1 103.3 SFN : LA : Gly in ratio LA : Gly 1:1 (freeze dried, SFN added 103.2 102.5 later*) SFN : Bet : Suc : Phe : MA : Gly in ratio Bet : Suc : Phe : MA 83.7 103.4 : Gly 0.4:0.1:0.1:2:0.6 SFN : Bet : Suc : Ect : CA : Gly in ratio Bet : Suc : Ect : CA 89.7 95.1 : Gly 0.25:0.1:0.1:1:1 *LA (in a water solution containing 90% LA, w / w) and Gly were mixed in a 1:1 molar ratio, followed by freeze-drying of the mixture before the addition of SFN). NA; Not analysed SFN retention (%) after 28 days at room T Formulation SFN loading 40% 60% SFN : LA : Gly in ratio LA : Gly 1:1 (60% freeze dried*) 46.4 89.2 SFN : Ty: C18:2 in ratio Ty : C18:21:1 54.8 59.7 SFN : TA : Gly in ratio TA : Gly 1:3 NA 94.9 SFN : CA : Gly in ratio CA : Gly 1:1 / 45.1 SFN : LA : Gly in ratio LA : Gly 1:1 (freeze dried, SFN 45.2 49.3 added later*) *LA (in a water solution containing 90% LA, w / w) and Gly were mixed in a 1:1 molar ratio, followed by freeze-drying of the mixture before the addition of SFN). NA: not analysed / : measurement not possible FTIR spectroscopy was performed for some formulations to further explore SFN stability in mixtures of two or more components that may form a eutectic or eutectic-like mixture or a DES. Spectra of neat SFN, formulations, particular combinations of components, etc. Briefly, SFN can act as hydrogen bond acceptor (N=C=S and S=O). Shifting in the peak position compound (in some solvent), compared to the peak of a pure compound, is generally marker for the hydrogen bond formation. The results are shown in the table below. Peak wavenumber (+ shifts d in DES) neat SFN + SFN + CA:Gly SFN + SFN + SFN CA:Gly 1:3 1:2.5 Ty:C18:21:1 Ty:C81:3 stretching 2177 2185 (d=8) 2184 (d=7) 2187 (d=10) 2190 (d=13) of N=C=S functional group 2097 2109 (d=12) 2110 (d=13) 2098 (d=1) 2110 (d=13) stretching 1019 1034 (d=15) 1039 (d=20) 999 (d=-20) 995 (d=-24) vibration of S=O The above-shown results indicate a solvent induced wavenumber shifts in the table imply the solute-solvent interaction. This suggests that the higher stability of SFN in tested formulations is due to formation of hydrogen bonds. Furthermore, some prepared formulations, namely Me:C18:2 (1:1), Ty:C18:2 (1:3), MA:Gly (1:3), LA:Gly (1:3), TA:Gly (1:3), CA:Gly (1:3), Bet:Ect:PG:C18:2 (1:1:15:15), Bet:Suc:Phe:MA:Gly (0.4:0.1:0.1:2:0.6) sndBet:Suc:Ect:CA:Gly (0.25:0.1:0.1:1:1) were tested for their biocompatibility at three different concentrations (500 mg / L, 1000 mg / L, and 2000 mg / L) on two human adherent cell lines: cancer cells derived from the colorectal adenocarcinoma (Caco-2 – ATCC No. HTB-37™) and normal human keratinocyte cells (HaCaT – CVCL No. 0038), to simulate conditions during topical or oral use. Cells were maintained in DMEM – Dulbecco's modified Eagle's medium (Capricorn Scientific GmbH), supplemented with 5% (v / v) FBS – fetal bovine serum (GIBCO by Life Technologies) in an incubator with 5% CO2and a humidified atmosphere at 37 °C. The impact of synthesized DESs on cellular proliferation was investigated in vitro utilizing the CellTiter96® AQueous One Solution Cell Proliferation assay (Promega), also known as the MTS assay. Briefly, cells were seeded in 96-well plates at a density of 3 × 104cells per well in 100 μl of media. Following overnight incubation, cells were exposed to the prepared DES at three concentrations spanning from 500 to 1000, and ultimately 2000 mg / L, and then incubated for 72 hours. A 10 μl volume of MTS reagent was added to each well and the absorbance was measured at 492 nm on the microplate reader after the 3-hour incubation period. Cell viability percentage was calculated by comparing the absorbance of treated cells to that of untreated control cells. The experiments were conducted in triplicate with four replicates for each concentration. The corresponding EC50values, defined as the concentration of tested compounds leading to 50% growth inhibition, were calculated from the dose–response curves. The EC50 values were determined to be greater than 2000 mg / L, suggesting that the tested DES exhibit no or low cytotoxicity. Moreover, all DES showed a slight stimulatory effect on cell proliferation at concentrations up to 1000 mg / L, while for HaCaT cells, the two hydrophobic DES (Me:C18:2 1:1 and Ty:C18:21:3) exhibited a pronounced stimulatory effect at 2000 mg / L, with cell viability reaching up to 160%. These findings suggest that the tested DES could be promising candidates for applications in the cosmetic and / or pharmaceutical sectors. The method for preparation of the above-formulations and thus for stabilizing SFN comprises the steps of: a) preparing a combination of at least two components selected in the group comprising: o organic acids, preferably selected in the group comprising decanoic acid, octanoic acid, linoleic acid, citric acid, lactic acid, malic acid, tartaric acid, oleic, other omega3 acids (such as (EPA), (DHA), a-linoleic (ALA)), omega6 acids (such as arachidonic (ARA), gamma-linoleic (GLA), dihomo-gamma-linoleic (DGLA)) acid, o amino acids, preferably selected in the group comprising proline and phenylalanine, o ectoine and 5-hydroxyectoine, o compounds comprising a tetramethyl ammonium moiety, preferably selected in the group comprising betaine, sarcosine, choline, carnitine, trimethylamine N-oxide (TMAO), glycerophosphorylcholine (GPC) and phosphatidylcholine (PC) and their salts, o phenols, o glycols, i.e. compounds having two hydroxyl (―OH) groups attached to different carbon atoms, preferably ethylene glycol, propylene glycol, butylene glycol, polyethylene glycol (PEG), o glycerol, o sugar alcohols, such as sorbitol, xylitol and similar compounds, o simple saccharides, preferably selected in the group comprising glucose, fructose, sorbose, sucrose, trehalose, o terpenoids, such as camphor, menthol, and thymol, o chaotropic agents, preferably selected in the group comprising urea, thiourea, guanidine, arginine and their salts, b) mixing the prepared combination and sulforaphane at the desired concentration, c) optionally adding further components, and d) storing at any temperature, preferably higher than 0 °C.

Claims

Patent claims 1. A formulation comprising an isothiocyanate (ITC) with a sulfoxide moiety and a combination of at least two of the following compounds: − organic acids, − amino acids, − ectoine and 5-hydroxyectoine, − compounds comprising a tetramethyl ammonium moiety, − phenols, − glycols, − glycerol, − sugar alcohols, − simple saccharides, − terpenoids, − chaotropic agents, wherein the above-mentioned compounds may be mixed in any combination with the selected isothiocyanate.

2. The formulation according to any of the preceding claims, wherein the organic acid is selected in the group comprising decanoic acid, octanoic acid, linoleic acid, citric acid, lactic acid, malic acid, tartaric acid, oleic, other omega3 acids, such as EPA, DHA, a-linoleic, omega6 acids, such as arachidonic, gamma-linoleic, dihomo-gamma-linoleic acid.

3. The formulation according to any of the preceding claims, wherein the compounds comprising a tetramethyl ammonium moiety is selected in the group comprising betaine, sarcosine, choline, carnitine, trimethylamine N-oxide (TMAO), glycerophosphorylcholine (GPC) and phosphatidylcholine (PC) and their salts.

4. The formulation according to any of the preceding claims, wherein the amino acid is proline or phenylalanine.

5. The formulation according to any of the preceding claims, wherein the glycol is selected in the group comprising propylene glycol, ethylene glycol, polyethylene glycol (PEG), and butylene glycol.

6. The formulation according to any of the preceding claims, wherein simple saccharide is selected in the group comprising glucose, fructose, sorbose, sucrose, trehalose.

7. The formulation according to any of the preceding claims, wherein chaotropic agent is selected in the group comprising urea, thiourea, guanidine, arginine and their salts.

8. The formulation according to any of the preceding claims, wherein sugar alcohol is sorbitol or xylitol.

9. The formulation according to any of the preceding claims, wherein the terpenoid is selected in the group comprising camphor, menthol, and thymol.

10. The formulation according to any of the preceding claims, wherein the isothiocyanate is of natural, synthetic, biosynthetic or semi-synthetic origin.

11. The formulation according to any of the preceding claims, wherein the formulation is selected in the group consisting of: − ITC : Me : C18:2, − ITC : Me : C10 − ITC : Ty: C18:2, − ITC : Ty: C8, − ITC : Ty: C10, − ITC : Pro: C8, − ITC : Pro : C18:2 : C8, − ITC : Cam : PA, − ITC : Me : Cam, − ITC : Ty : PG : C18:2, − ITC : Bet : PG, − ITC : Ect : PG, − ITC : PG : C18:2, − ITC : Bet : Ect : PG : C18:2, − ITC : Glc : CA, − ITC : LA : Gly, − ITC : LA : Pro, − ITC : Bet : LA, − ITC : Bet : LA:Gly,− ITC : Bet : LA : U, − ITC: Ect : LA, − ITC: Ect : LA : Gly, − ITC : MA : Gly, − ITC : CA : Gly, − ITC : TA : Gly, − ITC : Bet : Ect : LA : Gly, − ITC : Bet : Ect : MA : Gly, − ITC : Bet : Ect : CA : Gly, − ITC : Bet : Ect : TA : Gly, − ITC : Bet : Suc : Pro : LA : Gly, − ITC : Bet : Suc : Phe : MA : Gly, − ITC : Bet : Suc : Ect : CA : Gly, − ITC : Bet : Treh : Phe : MA : Gly, and − ITC : Bet : Treh : Ect : LA : Gly.

12. The formulation according to any of the preceding claims, wherein it comprises additional components.

13. The formulation according to any of the preceding claims, wherein the molar ratio of each component of the combination with respect to the total moles of the combination may range from 0.01 to 0.

9.

14. The formulation according to any of the preceding claims, wherein the isothiocyanate is sulforaphane (SFN) or sulforaphene (SFEN).

15. The formulation according to any of the preceding claims, wherein the combination of said compounds to be mixed with SFN or SFEN forms a eutectic-like mixture.

16. The formulation according to the preceding claim, wherein the eutectic-like mixture is a deep eutectic system.

17. The formulation according to any claim from 13 to 16, wherein the formulation is selected in the group consisting of: − SFN : Me : C18:2,− SFN : Me : C10 − SFN : Ty: C18:2, − SFN : Ty: C8, − SFN : Ty: C10, − SFN : Pro: C8, − SFN : Pro : C18:2 : C8, − SFN : Cam : PA, − SFN : Me : Cam, − SFN : Ty : PG : C18:2, − SFN : Bet : PG, − SFN : Ect : PG, − SFN : PG : C18:2, − SFN : Bet : Ect : PG : C18:2, − SFN : Glc : CA, − SFN : LA : Gly, − SFN : LA : Pro, − SFN : Bet : LA, − SFN : Bet : LA:Gly, − SFN : Bet : LA : U, − SFN : Ect : LA, − SFN : Ect : LA : Gly, − SFN : MA : Gly, − SFN : CA : Gly, − SFN : TA : Gly, − SFN : Bet : Ect : LA : Gly, − SFN : Bet : Ect : MA : Gly, − SFN : Bet : Ect : CA : Gly, − SFN : Bet : Ect : TA : Gly, − SFN : Bet : Suc : Pro : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, − SFN : Bet : Suc : Ect : CA : Gly, − SFN : Bet : Treh : Phe : MA : Gly, and − SFN : Bet : Treh : Ect : LA : Gly.

18. The formulation according to the preceding claim, wherein the formulation is selected in the group consisting of following formulations with given molar ratios: SFN + additional components molar ratio of additional components Me: C18:2 1:1 Me : C18:2 1:3 Me : C18:2 1:5 Me :C8 1:1 Ty : C18:2 1:1 Ty : C18:2 1:3 Ty : C18:2 1:5 Ty :C8 1:3 Ty : C10 1:1 Pro : C8 1:4 Pro : C18:2 : C8 1:4:2 Cam : PA 1:1 Me : Cam 1:1 Ty : PG: C18:2 1:1:1 Bet : PG 1:4 Ect : PG 1:12 PG : C18:2 1:1 Ect : PG: C18:2 1:12:6 Bet : Ect : PG : C18:2 1:1:15:6 Bet : Ect : PG : C18:2 1:1:15:15 Glc : CA 1:1:6.5(H2O)LA : Pro 2:1:1(H2O)Bet : LA : U 1:2:0.5:1(H2O)Ect : LA 1:2:1(H2O)Glc : LA 1:2:1(H2O)LA : Gly 1:1:1(H2O)LA : Gly 1:2:1(H2O)Bet : LA 1:4:0.5(H2O)Bet : LA : Gly 1:1:2:0.5(H2O)Ect : LA : Gly 1:3:2:2(H2O)Bet : Ect : LA : Gly 0.5:0.5:3:2:2(H2O)MA : Gly 1:2.5:2(H2O)MA : Gly 1:3 CA : Gly 1:1:1.5(H2O)CA : Gly 1:2.5:2(H2O)CA : Gly 1:3 TA : Gly 1:2.5:2(H2O)TA : Gly 1:3 Ect : Bet : MA : Gly 0.5:0.5:1:2:2(H2O)Ect : Bet : CA : Gly 0.5:0.5:1:2:2(H2O)Ect : Bet : TA : Gly 0.5:0.5:1:3:3(H2O)Bet : Suc : Pro : LA : Gly 0.4:0.1:0.1:2:0.5:3(H2O)Bet : Suc : Phe : MA : Gly* 0.4:0.1:0.1:2:0.6:4(H2O)Bet : Suc : Phe : MA : Gly* 0.4:0.1:0.1:2:5 Bet : Treh : Phe : MA : Gly 0.4:0.1:0.1:2:0.6:4(H2O)Bet : Tre : Ect : LA : Gly 0.4:0.1:0.1:2:0.6:1(H2O)Bet : Suc : Ect : CA : Gly 0.25:0.1:0.1:1:1:2.5(H2O)19. The formulation according to any of the preceding claims, wherein the isothiocyanate is stable at 4 and / or 28 °C and the formulation is selected in the group consisting of: − SFN : Me : C18:2, − SFN : Me : C10, − SFN : Ty : C18:2, − SFN : Ty : C8, − SFN : Ty : C10, − SFN : Me : Cam, − SFN:Ty:PG:C18:2, − SFN : Ect : PG, − SFN : Bet : Ect : PG: C18:2, − SFN : Glc : CA, − SFN : LA : Gly, − SFN : Bet : LA : Gly, − SFN : MA : Gly, − SFN : CA : Gly,− SFN : TA : Gly, − SFN : Bet : Ect : TA : Gly, − SFN : Bet : Suc : Pro : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, − SFN : Bet : Suc : Ect : CA : Gly, − SFN : Bet : Treh : Phe : MA : Gly, and − SFN : Bet : Treh : Ect : LA : Gly.

20. The formulation according to the preceding claim, wherein the formulation is prepared in the following ratios and the SFN concentration is arbitrary, preferably up to 60% (w / w): − SFN : Me : C18:2 in ratio Me : C18:21:1, 1:3 or 1:5, − SFN : Me : C10 in ratio Me : C101:1, − SFN : Ty : C18:2 in ratio Ty : C18:21:3, 1:5 or 1:7, − SFN : Ty : C8 in ratio Ty : C8 1:3, − SFN : Ty : C10 in ratio Ty : C101:1, − SFN : Me : Cam in ratio Me : Cam 1:1, − SFN : Ty : PG : C18:2 in ratio Ty : PG : C18:21:1:1, − SFN : Ect : PG in ratio Ect : PG 1:12, − SFN : Bet : Ect : PG : C18:2 in ratio Bet : Ect : PG : C18:21:1:15:15, − SFN : LA : Gly in ratio LA : Gly 1:1, or 1:2, − SFN : M A : Gly in ratio MA : Gly 1:2.5 or 1:3, − SFN : CA : Gly in ratio CA : Gly 1:1, 1:2.5, or 1:3, − SFN : TA : Gly in ratio TY : Gly 1:2.5 or 1:3, − SFN : Bet : Ect : TA : Gly in ratio Bet : Ect : TA : Gly 0.5:0.5:1:3, − SFN : Bet : Suc : Pro : LA : Gly in ratio Bet : Suc : Pro : LA : Gly 0.4:0.1:0.1:2:0.5, − SFN : Bet : Suc : Phe : MA : Gly in ratio Bet : Suc : Phe : MA : Gly 0.4:0.1:0.1:2:0.6, − SFN : Bet : Suc : Ect : MA : Gly in ratio Bet : Suc : Ect : MA : Gly 0.4:0.1:0.1:2:5, − SFN : Bet : Suc : Ect : CA : Gly in ratio Bet : Suc : Ect : CA : Gly 0.25:0.1:0.1:1:1, − SFN : Be : Treh : Phe : MA : Gly in ratio Bet : Treh : Phe : MA : Gly 0.4:0.1:0.1:2:0.6, − SFN : Bet : Treh : Ect : LA : Gly in ratio Bet : Treh : Ect : LA : Gly 0.4:0.1:0.1:2:0.

6.

21. The formulation according to any of the preceding claims, wherein the isothiocyanate is stable for at least 60 days and the formulation is selected in the group consisting of: − SFN : Me : C18:2,− SFN : Me : C10, − SFN : Ty : C18:2, − SFN : LA : Gly − SFN : MA : Gly, − SFN : CA : Gly, − SFN : TA:Gly, − SFN : Bet : Suc : Pro : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, − SFN : Bet : Suc : Ect : CA : Gly, − SFN : Bet : Treh : Phe : MA : Gly, and − SFN : Bet : Treh : Ect : LA : Gly.

22. The formulation according to the preceding claim, wherein the formulation is selected in the group of formulations prepared in the following ratios and the SFN concentration is arbitrary, preferably up to 60% (w / w): − SFN : Me : C18:2, wherein the molar ratio of Me and C18:2 is preferably 1:1, 1:3 or 1:5, − SFN : Me : C10, − SFN : Ty : C18:2, wherein the molar ratio of Ty and C18:2 is preferably 1:3, 1:5 or 1:7, − SFN : LA : Gly, wherein the molar ratio of LA and Gly is preferably 1:1 or 1:2, − SFN : MA : Gly, wherein the molar ratio of MA and Gly is preferably 1:2.5 or 1:3, − SFN : CA : Gly, wherein the molar ratio of CA and Gly is preferably 1:1; 1:2.5 or 1:3, − SFN : TA : Gly, wherein the molar ratio of TA and Gly is preferably 1:2.5 or 1:3, − SFN : Bet : Suc : Pro : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, in ratio Bet:Suc:Phe:MA:Gly 0.4:0.1:0.1:2:0.6 − SFN : Bet : Suc : Ect : CA : Gly, in ratio Bet:Suc:Ect:CA:Gly 0.25:0.1:0.1:1:1, − SFN : Bet : Treh : Phe : MA : Gly, in ratio Bec:Suc:Ect:MA:Gly 0.4:0.1:0.1:2:5 and − SFN : Bet : Treh : Ect : MA : Gly. Bet:Treh:Phe:MA:Gly 0.4:0.1:0.1:2:0.

6.

23. The formulation according to any of the preceding claims, wherein it comprises up to 60% isothiocyanate, preferably up to 60% of sulforaphane.

24. The formulation according to the preceding claims, wherein the concentration of SFN is from 20 to 60% (w / w).

25. The formulation according to any of the preceding claims, wherein the concentration of SFN is from 20 to 60% (w / w), preferably 20%, 40% or 60% (w / w) and the SFN is stable at temperatures higher than 0 C, wherein the formulation is selected in the group comprising: − SFN : LA : Gly, − SFN : Ty : C18:2, − SFN : MA : Gly, − SFN : TA : Gly, − SFN : CA : Gly, − SFN : LA : Gly, − SFN : Bet : Suc : Phe : MA : Gly, − SFN : Bet : Suc : Ect : CA : Gly.

26. The formulation according to the preceding claim, wherein it is selected in the group consisting of formulations prepared in the following ratios and the SFN concentration is arbitrary, preferably up to 60% (w / w): − SFN : LA : Gly in ratio LA : Gly 1:1, − SFN : Ty : C18:2 in ratio Ty : C18:2, − SFN : MA : Gly in ratio MA : Gly 1:3, − SFN : TA : Gly in ratio TA : Gly 1:3, − SFN : CA : Gly in ratio CA : Gly 1:1, − SFN : LA : Gly in ratio LA : Gly 1:1, − SFN : Bet : Suc : Phe : MA : Gly in ratio Bet : Suc : Phe : MA : Gly 0.4:0.1:0.1:2:0.6, − SFN : Bet : Suc : Ect : CA : Gly in ratio Bet : Suc : Ect : CA : Gly 0.25:0.1:0.1;1:

1.

27. The formulation according to any of the preceding claims, wherein the formulation is a pharmaceutical preparation, a food or drink product, a nutraceutical product, a dietary supplement, a food additive, a cosmetic product, a skin product, a hair product, a feed additive, or an agricultural product.

28. A pharmaceutical preparation, a food or drink product, a nutraceutical product, a dietary supplement, a food additive, a cosmetic product, a skin product, a hair product, a feed additive or an agricultural product comprising the formulation according to any of the preceding claims,29. A method for preparation of the formulation comprising an isothiocyanate according to any of the preceding claims comprises the steps of: i. preparing a combination of at least two components selected in the group consisting of: − organic acids, − amino acids, − ectoine and 5-hydroxyectoine, − compounds comprising a tetramethyl ammonium moiety, − phenols, − glycols, − glycerol, − sugar alcohols, − simple saccharides, − terpenoids, − chaotropic agents, ii. mixing the prepared combination and the selected isothiocyanate, preferably sulforaphane or sulforaphene, and iii. optionally adding any further components.

30. The method according to the preceding claim, wherein a step of water removal is performed either before or after step ii.

31. The method according to claim 29 or 30, wherein the prepared combination or the prepared combination mixed with ITC, preferably SFN, is freeze-dried.

32. A method for stabilizing an isothiocyanate, characterized in that the method comprises the steps of: a) preparing a combination of at least two components selected in the group consisting of: − organic acids, − amino acids, − ectoine and 5-hydroxyectoine, − compounds comprising a tetramethyl ammonium moiety, − phenols, − glycols,− glycerol, − sugar alcohols, − simple saccharides, − terpenoids, − chaotropic agents, b) mixing the prepared combination and the selected isothiocyanate, preferably sulforaphane or sulforaphene at the desired concentration, wherein water removal may optionally be performed before or after step b), c) optionally adding further components, and d) storing at any temperature, preferably higher than 0 °C.

33. Use of the method and / or formulation according to any of the preceding claims in preparation of formulations, products or any other components.

34. Use according to claim 33 in preparation of a pharmaceutical product, a food or drink product, a nutraceutical product, a dietary supplement, a food additive, a cosmetic product, a skin product, a hair product, a feed additive or an agricultural product.

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