Isothiocyanate derivative

Abstract

To provide isothiocyanates (ITCs) or derivatives thereof which are stable in an aqueous solution and also stable to heat.SOLUTION: There is provided a compound of the formula or a pharmaceutically acceptable salt thereof. In the formula, R1, R2, and R3 are an acetyl group or H, R4 is OH or amino acid of 1 to 3 residues, and R5 is H or amino acid of 1 to 3 residues. The present invention provides: isothiocyanate derivatives caused to be stable in an aqueous solution and also during heating them by reacting isothiocyanates (ITCs) derived from Moringa with a peptide having a cysteine group such as glutathione; and a production method thereof.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an isothiocyanate derivative or a pharma- ceutically acceptable salt thereof, in particular to an isothiocyanate derivative or a pharma- ceutically acceptable salt thereof derived from Moringa, and a method for preparing the same. [Background technology]

[0002] Moringa is the common name for the Moringa oleifera, a tree of the Moringaceae family, and is a medicinal plant native to northwestern India. In recent years, it has also been attracting attention in Japan as a superfood due to its high nutritional value and rich content of a wide range of nutrients. Moringa is not only highly nutritious, but is also said to have physiological functions such as antioxidant effects, anti-inflammatory effects, antihypertensive effects, antispasmodic effects, and cholesterol-reducing effects, and its use as a health food is recommended worldwide as it is useful for preventing disease and maintaining health.

[0003] In particular, isothiocyanates (ITCs) derived from Moringa are less volatile and more chemically stable than isothiocyanates derived from the Brassicaceae family, and the present applicant has found that they suppress fat accumulation in the liver and have an anti-obesity effect in mouse experiments (2019 Annual Meeting of the Agricultural Chemical Society of Japan). ITCs are produced when glucosinolates contained in Moringa react with endogenous myrosinase and are enzymatically decomposed when cells are destroyed during the process of crushing or solvent extraction. However, ITCs are unstable in aqueous solution and have poor thermal stability in aqueous solution, so the content of ITCs produced decreases when the product is sterilized by heating. Summary of the Invention [Means for solving the problem]

[0004] The present invention provides an isothiocyanate derivative that is stable in an aqueous solution or when heated, and a method for producing the same, by reacting moringa-derived isothiocyanates (ITCs) with a peptide having a cysteine ​​group, such as glutathione.

[0005] Thus, the present invention provides the following: (Item 1) formula [ka] or a pharma- ceutically acceptable salt thereof, wherein R1, R2 and R3 are acetyl groups or H; R4 is OH or an amino acid having 1 to 3 residues; R5 is H or 1 to 3 amino acid residues; A compound, or a pharma- ceutically acceptable salt thereof, is provided. (Item 2) R1, R2 and R3 are H or R1 is an acetyl group, and R2 and R3 are H; or R2 is an acetyl group and R1 and R3 are H; or R3 is an acetyl group, R1 and R2 are H; R4, [ka] (where A represents the point of attachment) and R5, [ka] (where A represents the point of attachment) That is, A compound according to any one of the preceding items, or a pharma- ceutically acceptable salt thereof. (Item 3) R1, R2 and R3 are H or R1 is an acetyl group, and R2 and R3 are H; or R2 is an acetyl group and R1 and R3 are H; or R3 is an acetyl group, R1 and R2 are H; R4 is OH; R5 is H; A compound according to any one of the preceding items, or a pharma- ceutically acceptable salt thereof. (Item 4) R1, R2 and R3 are H or R1 is an acetyl group, and R2 and R3 are H; or R2 is an acetyl group and R1 and R3 are H; or R3 is an acetyl group, R1 and R2 are H; R4 is OH; R5 [ka] (where A represents the point of attachment) 8. The compound of any one of the preceding items, wherein: (Item 5) 2. A method for making the compound of any one of the preceding claims, or a pharma- ceutically acceptable salt thereof, comprising reacting an extract of a plant of the Moringa family and a yeast extract containing cysteine ​​peptides in an aqueous medium. (Item 6) Item 11. The method of any one of the preceding items, wherein the cysteine ​​peptide is glutathione. (Item 7) 13. The method of any one of the preceding items, wherein the extract of a plant of the Moringa family is a Moringa extract. (Item 8) 10. The method of any one of the preceding items, wherein the plant extract of the Moringaceae family comprises moringin and / or an acetylated form of moringin. (Item 9) 2. The method of any one of the preceding items, wherein the amount of yeast extract added comprises about 10% by weight or more relative to the amount of plant extract of the Moringa family added. (Item 10) 10. The method according to any one of the preceding claims, wherein the yeast extract is a yeast extract from Torula yeast or a yeast extract from Saccharomyces cerevisiae. (Item 11) 13. The method of any one of the preceding items, further comprising the step of adding dextrin. (Item 12) 2. The method of any one of the preceding items, wherein the amount of dextrin added comprises about 20% by weight or more relative to the amount of the plant extract of the Moringa family added. (Item 13) 2. The method of any one of the preceding items, further comprising the step of heat sterilizing the aqueous solution obtained by the reacting step at about 65° C. or higher. (Item 14) A composition comprising the compound according to any one of the preceding items or a pharma- ceutically acceptable salt thereof, which has anti-obesity, weight gain suppression, lipid metabolism improvement, liver function improvement, and / or fatty liver prevention / improvement effects.

[0006] It is contemplated that one or more of the above features may be provided in combinations other than those specifically described. Still further embodiments and advantages of the present invention will be recognized by those skilled in the art upon reading and understanding the following detailed description, if necessary.

[0007] Furthermore, other features and notable actions and effects of the present invention besides those described above will become apparent to those skilled in the art by referring to the following detailed description of the preferred embodiments of the present invention and the drawings. Effect of the Invention

[0008] The present invention provides a stable isothiocyanate derivative having useful physiological functions, which can provide a functional material that is useful for preventing diseases and supporting healthy longevity. [Brief description of the drawings]

[0009] [Figure 1]FIG. 1 shows the procedure for producing powdered ITCs in Example 1. [Diagram 2] FIG. 2 is a graph showing the remaining amount of each isothiocyanate derivative after heating when cysteine, oxidized glutathione (GSSG), or reduced glutathione (GSH) was added as a stabilizer for isothiocyanate in one embodiment of the present invention. [Diagram 3] FIG. 3 is a graph showing the results of investigating the amount of yeast extract added in one embodiment of the present invention. [Figure 4] FIG. 4 is a graph showing the results of investigating the extraction temperature in one embodiment of the present invention. [Diagram 5] FIG. 5 is a graph showing the results of examining extraction time in one embodiment of the present invention. [Figure 6] FIG. 6 is a graph showing the results of examining the type of filter aid in one embodiment of the present invention. [Figure 7] FIG. 7 is a graph showing the results of examining the sterilization mode in one embodiment of the present invention. [Figure 8] FIG. 8 is a graph showing the results of examining the powderization process in one embodiment of the present invention. [Figure 9] FIG. 9 is a graph showing the anti-obesity effect of an isothiocyanate derivative in one embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] The following provides an explanation of the definitions of terms particularly used in this specification and / or basic technical content as appropriate. In addition, an embodiment and an example of the present invention will be described with reference to the drawings.

[0011] As used herein, "about" means ±10% of the preceding numerical value.

[0012] As used herein, a "derivative" refers to a compound or portion of a compound that is derived from or theoretically derivable from a parent compound.

[0013] As used herein, the term "isothiocyanate" refers to a compound having an isothiocyanate group (-NCS group).

[0014] As used herein, the term "cysteine ​​peptide" refers to a tripeptide consisting of the three amino acids glutamic acid, cysteine, and glycine, and includes, for example, glutathione.

[0015] As used herein, "anti-obesity" refers to the promotion of reduction in body fat and / or inhibition of accumulation of body fat, thereby relieving or preventing obesity. In addition, in the present invention, "obesity" refers to a state in which fat is excessively accumulated in adipose tissue in the body, and includes, for example, a body mass index (BMI) of 25 or more.

[0016] In this specification, the term "suppression of weight gain" is used to mean not only suppression of weight gain, but also maintenance or reduction of weight. In addition, suppression of weight gain also includes slimming. The degree of suppression of weight gain can be evaluated using body weight and BMI as indicators.

[0017] In this specification, "improving lipid metabolism" means, in lipid metabolism, decomposing (promoting decomposition) lipids and their components (fatty acids) present in body tissues, blood, and lymph (lymphatic tissue), preventing or suppressing the accumulation of lipids and their components in body tissues (particularly, adipose tissue), and reducing lipids and their components accumulated in body tissues (particularly, adipose tissue). Specifically, the term "improving lipid metabolism" includes preventing or suppressing the high concentration of lipids and their components ingested by eating in blood (particularly, serum) and lymph (lymphatic tissue), reducing lipids present at high concentrations in blood and lymph (lymphatic tissue), preventing or suppressing the accumulation of lipids and their components ingested by eating in body tissues, decomposing or reducing lipids and their components accumulated in body tissues, or decomposing lipids and their components present in blood (particularly, serum) or reducing their concentrations when lipids and their components accumulated in body tissues are released into the blood.

[0018] As used herein, "improving liver function" means improving liver function, and for example, the function can be indicated by an indicator (e.g., AST: aspartate aminotransferase). For example, "improving liver function" includes lowering or alleviating an increase in blood AST levels in individuals with high blood AST levels (including those who are normal but have high levels), maintaining blood AST levels in individuals with normal or low blood AST levels, and preventing or alleviating an increase in blood AST levels.

[0019] In this specification, "fatty liver prevention and improvement" means that fatty liver is prevented and / or improved. The blood ALT concentration is a value used as an index of liver function, and can be used to evaluate the effect of preventing and / or improving fatty liver. In addition, since the blood ALT concentration increases in fatty liver, the blood ALT concentration may increase as fat accumulation, such as visceral fat, increases. For this reason, the blood ALT concentration can also be used as a measure of visceral fat mass. In this specification, the term "fatty liver" is defined as a state of excessive accumulation of fat in liver cells caused by lipid metabolism disorder, where fat reaches 5% or more of the liver weight. This causes various diseases such as angina pectoris, myocardial infarction, stroke, arteriosclerosis, fatty liver, and pancreatitis. Fatty liver is divided into alcoholic fatty liver caused by alcohol consumption and non-alcoholic fatty liver disease (NAFLD) not caused by alcohol.

[0020] In one aspect of the invention, the formula [ka] or a pharma- ceutically acceptable salt thereof, wherein R1, R2 and R3 are acetyl groups or H; R4 is OH or an amino acid having 1 to 3 residues; R5 is H or 1 to 3 amino acid residues; The present invention provides, without limitation, a compound, or a pharma- ceutically acceptable salt thereof.

[0021] In one embodiment, R1, R2 and R3 are H or R1 is an acetyl group, and R2 and R3 are H; or R2 is an acetyl group and R1 and R3 are H; or R3 is an acetyl group, R1 and R2 are H; R4, [ka] (where A represents the point of attachment) and R5, [ka] (where A represents the point of attachment) That is, A compound, or a pharma- ceutically acceptable salt thereof, is provided.

[0022] In one embodiment, R1, R2 and R3 are H or R1 is an acetyl group, and R2 and R3 are H; or R2 is an acetyl group and R1 and R3 are H; or R3 is an acetyl group, and R1 and R2 are H; A compound, or a pharma- ceutically acceptable salt thereof, is provided.

[0023] In one embodiment, a compound is provided, wherein R4 is OH and R5 is H, or a pharma- ceutically acceptable salt thereof.

[0024] In one embodiment, R1, R2 and R3 are H or R1 is an acetyl group, and R2 and R3 are H; or R2 is an acetyl group and R1 and R3 are H; or R3 is an acetyl group, R1 and R2 are H; R4 is OH; R5, [ka] (where A represents the point of attachment) That is, A compound, or a pharma- ceutically acceptable salt thereof, is provided.

[0025] In one aspect of the invention, the formula [ka] or a pharma- ceutically acceptable salt thereof, wherein R1, R2 and R3 are H or R1 is an acetyl group, and R2 and R3 are H; or R2 is an acetyl group and R1 and R3 are H; or R3 is an acetyl group, and R1 and R2 are H; A compound, or a pharma- ceutically acceptable salt thereof, is provided.

[0026] In another aspect of the present invention, there is provided a method for producing such a compound or a pharma- ceutically acceptable salt thereof, comprising reacting a plant extract of the Moringa family and a yeast extract containing a cysteine ​​peptide in an aqueous medium.Preferably, the cysteine ​​peptide is glutathione.

[0027] In one embodiment, the plant extract of the family Moringaceae may be any, for example, a Moringa extract may be used. The Moringa extract may be obtained by extracting Moringa using various solvents by known methods. The whole plant or any part of the plant may be crushed, for example, by a blender, and then a solvent extract may be obtained using an extraction solvent. Moringa may be dried before crushing and then crushed. The solvent extract may also be produced in a powder state by additional processes such as vacuum distillation and freeze-drying or spray-drying. The Moringa powder may also be obtained by crushing Moringa using a known crusher or mill.

[0028] In one embodiment, a method for making such a compound or a pharma- ceutically acceptable salt thereof comprises reacting Moringa leaves and yeast extract containing glutathione in water. Preferably, the cysteine ​​peptide is glutathione.

[0029] The moringa to be extracted or pulverized is not particularly limited, but examples thereof include Moringa oleifera, Moringa concanensis, Moringa drouhardii, etc. Moringa oleifera is preferably used because it is widely cultivated and can be easily harvested.

[0030] The parts of moringa that can be extracted or crushed include leaves, stems, pods (flesh), and seeds. These parts may be used raw or after drying. From the viewpoint of storage stability as a raw material and yield in producing an extract, dried moringa is preferably used.

[0031] The solvent used for extraction is not particularly limited as long as it can extract an extract from Moringa, and for example, water, an organic solvent, or a mixed solvent of water and an organic solvent can be used. From the viewpoint of safety, it is preferable to extract with water only. Examples of organic solvents include lower alcohols that can be mixed with water (monohydric or polyhydric alcohols having C1 to C4, such as methanol, ethanol, propanol, propylene glycol, butylene glycol, and glycerin), acetone, benzene, hexane, diethyl ether, and dichloromethane. When using a mixed solvent of these organic solvents and water, they may be mixed with water before use, or two or more of them may be mixed with water and used. When using a mixed solvent of an organic solvent and water as a solvent, the mixing ratio of the organic solvent may be more than 0% to less than 100%. The amount of the solvent used for extraction is not particularly limited, but can be appropriately set depending on the type of extraction solvent used. For example, the amount can be about 1 to about 20 times, or about 5 to about 20 times, more preferably about 5 to about 10 times, and most preferably about 5 to about 8 times, based on the dry weight of the moringa to be extracted. The temperature of the solvent during extraction is not particularly limited, but can be, for example, about 20 to about 95°C. The extraction time is not particularly limited, but can be, for example, about 30 to about 150 minutes from the viewpoint of production efficiency. Extraction can be performed under stirring or in a stationary state. After extraction, a process such as filtration or centrifugation is performed to remove residues, and the extraction solvent can then be removed by reducing pressure or the like. In addition, when the extract is made into a powder, a dried extract obtained by distilling the solvent extract under reduced pressure and then freeze-drying or spray-drying can be produced as necessary.

[0032] In other embodiments, Moringa extracts can be obtained using extraction processes known in the art, such as digestion, infusion, percolation, digestion, decoction, high temperature continuous extraction, aqueous-alcoholic extraction, countercurrent extraction, microwave assisted extraction, ultrasonic extraction, subcritical fluid extraction, tissue piece extraction, etc., either alone or in combination.

[0033] In the method for producing the compound of the present invention, each substance may be derived from a natural product or may be prepared by a known method. Each raw material compound may be used as a salt. The compound obtained in each step of the method of the present invention may be used in the next reaction as a reaction solution or as a crude product, or may be isolated and purified from the reaction solution by known separation and purification means, such as concentration, vacuum concentration, solvent extraction, crystallization, recrystallization, transfer, chromatography, etc., and used in the next reaction. When the compound is commercially available, the commercially available product may be used as it is.

[0034] In one embodiment, the compound of the present invention is an isothiocyanate derivative obtained by covalently bonding a peptide having a cysteine ​​group to isothiocyanates (ITCs) derived from Moringa via a dithiocarbamate structure (herein referred to as the "compound of the present invention" or the "isothiocyanate derivative of the present invention").

[0035] In one embodiment, the compound of the present invention is an isothiocyanate derivative obtained by covalently bonding glutathione to isothiocyanates (ITCs) derived from Moringa via a dithiocarbamate structure (herein referred to as the "compound of the present invention" or the "isothiocyanate derivative of the present invention").

[0036] In Moringa, isothiocyanates are generated from glycated precursor glucosinolates by the action of myrosinase (thioglucoside glucohydrolase, β-thioglucosidase), an enzyme that is activated upon wounding or digestion of plant tissue. Myrosinase cleaves the thio-linked glucose of glucosinolates to generate aglycones and subsequent formation of active ITCs.

[0037] Four types of isothiocyanates derived from Moringa are known (compounds 5-8) obtained from the following Moringa glucosinolates (compounds 1-4) (Phytochemistry, Volume 103, July 2014, Pages 114-122). [ka]

[0038] Of these, compound 5 (4-[(α-L-rhamnosyloxy)benzyl]isothiocyanate) and compound 8 (4-[(4'-O-acetyl-α-L-rhamnosyloxy)benzyl]isothiocyanate) are the most abundant isothiocyanates formed from glucosinolates 1 and 4, and typically account for more than 95% of the total ITCs present. In contrast, compounds 6 (4-[(2'-O-acetyl-α-L-rhamnosyloxy)benzyl]isothiocyanate) and 7 (4-[(3'-O-acetyl-α-L-rhamnosyloxy)benzyl]isothiocyanate) are formed in small amounts from the precursors of the respective glucosinolates, compounds 2 and 3 (Phytochemistry, Volume 103, July 2014, Pages 114-122). Compound 5 is also called moringin, and compounds 6-8 are the acetylated forms of moringin.

[0039] In one embodiment, the compound of the present invention is obtained by covalently bonding a cysteine-containing compound to the isothiocyanate as described above via a dithiocarbamate structure. The bond between the isothiocyanate and the cysteine-containing compound is formed by reacting the isothiocyanate group of the isothiocyanate with the thiol group of the cysteine.

[0040] In one embodiment, the compound of the present invention is obtained by covalently binding glutathione to the isothiocyanate as described above via a dithiocarbamate structure. The bond between the isothiocyanate and glutathione is formed by reacting the isothiocyanate group of the isothiocyanate with the cysteine ​​group of the glutathione.

[0041] In one embodiment of the present invention, the glutathione that reacts with isothiocyanate is preferably derived from a yeast extract, and examples of such yeast extracts include those derived from fermentation products of the genus Saccharomyces, Schizosaccharomyces, Candida, etc. Among these, those derived from fermentation products of the genus Saccaromyces are preferred. Examples of those derived from fermentation products of the genus Saccaromyces include yeast extract and yeast extract with high γ-glutamylcysteine ​​content.

[0042] As the yeast extract that can be used in the production method of the present invention, yeast extracts derived from yeasts belonging to the genus Saccharomyces such as Saccharomyces cerevisiae, the genus Schizosaccharomyces such as Schizosaccharomyces pombe, and the genus Candida such as Candida utilis are usually used, and yeast extracts derived from yeasts belonging to the genus Saccharomyces or Candida are preferred. Preferably, yeast extracts derived from Torula yeast or yeast extracts derived from Saccharomyces cerevisiae are used.

[0043] The compound of the present invention can be produced by reacting a plant extract of the family Moringaceae and a yeast extract containing glutathione in an aqueous medium. Preferably, the compound can be produced by reacting moringa leaves and a yeast extract containing glutathione in an aqueous medium. For example, yeast extract is added to moringa leaves in an amount of usually about 5% by weight to about 30% by weight, preferably about 10% by weight to about 20% by weight, for example, at least about 10% by weight or more, about 15% by weight or more, about 20% by weight or more, and then water is added, for example, about 15 times the amount of water relative to the amount of moringa leaves, and the mixture is mixed and adjusted to a pH of usually about 6 to about 11, preferably pH 7 to 9 to obtain an aqueous solution. The amount of solvent (water) required relative to the weight of moringa is usually about 2.5 to about 20 times the amount of water, preferably about 2.5 to about 15 times the amount of water relative to the weight of moringa, and mixed. The resulting mixture (aqueous solution) may be adjusted to the above pH after adding yeast extract or the like.

[0044] Then, the mixture is left to stand or stirred at about 8 to about 40° C., preferably about 10 to about 25° C., for about 0.25 to about 12 hours, preferably about 1 to 6 hours. If necessary, a filter aid such as radiolite is added, and the mixture is filtered to obtain a reaction liquid. The product obtained by the reaction can be purified by a conventional method such as chromatography.

[0045] In one embodiment, dextrin such as dextrin, maltodextrin, cyclodextrin γ, cluster dextrin, etc. may be further added to the filtered reaction solution. In this case, the amount of dextrin added is usually about 5% by weight to about 70% by weight, preferably about 10% by weight to about 30% by weight, for example, about 10% by weight or more, about 15% by weight or more, about 20% by weight or more, based on the amount of the plant extract of the Moringa family added.

[0046] In one embodiment, in the method of the present invention, the reaction solution obtained by reacting the yeast extract containing the plant extract of the Moringa family and glutathione in an aqueous medium, the reaction solution obtained by filtering the reaction solution, or the reaction solution to which dextrin has been added can be heat sterilized. A known method can be used for heat sterilization, and the heating temperature and time may be such that the bacterial count test results in a negative result for Escherichia coli and / or the general viable bacterial count is about 3000 CFU or less. For example, heat sterilization can be performed at about 60°C to 150°C, preferably about 65°C to 130°C, for about 1 to about 30 minutes, preferably about 10 minutes, but the sterilization temperature and sterilization time can be appropriately set so as to pass the above-mentioned bacterial count test. In one embodiment, heat sterilization may be performed at about 90°C or higher for about 10 minutes or at about 121°C or higher for about 2 minutes, etc.

[0047] The aqueous medium to be used may be any medium that falls within the category known as an aqueous medium, such as distilled water, ion-exchanged water, industrial water, and tap water, and may further contain an organic solvent.

[0048] As the organic solvent, lower alcohols such as methanol, ethanol, propanol, isopropanol, etc., polyhydric alcohols such as 1,3-butanediol, propanediol, dipropanediol, glycerin, etc., ethers such as diethyl ether, dipropyl ether, etc., esters such as ethyl acetate, butyl acetate, etc., ketones such as acetone, ethyl methyl ketone, etc., chloroform, dichloromethane, acetonitrile, hexane, etc. can be used, and one or more of these can be selected and used. In addition, physiological saline, phosphate buffer, phosphate buffered physiological saline, etc. may be used. Among them, as the organic solvent, lower alcohols, 1,3-butanediol, glycerin, ethers, ethyl acetate, acetone, chloroform, dichloromethane, acetonitrile, and hexane are preferred, lower alcohols, ethers, etc. are more preferred, and methanol, ethanol, and diethyl ether are particularly preferred.

[0049] In one embodiment, the isothiocyanate derivative of the present invention can be used for anti-obesity, weight gain suppression, lipid metabolism improvement, liver function improvement, and / or fatty liver prevention. Therefore, in another aspect of the present disclosure, a composition for preventing or treating obesity, or having anti-obesity, weight gain suppression, lipid metabolism, liver function improvement, and / or fatty liver prevention and improvement effects, comprising the compound of the present invention or a pharma- ceutical acceptable salt thereof, is provided.

[0050] Another aspect of the present invention includes a method for preventing or treating obesity in a subject, or having anti-obesity, weight gain suppression, lipid metabolism improvement, liver function improvement, and / or fatty liver prevention effects, comprising administering to the subject an effective amount of a compound of the present invention or a pharma- ceutical acceptable salt thereof.

[0051] The compound of the present invention can also be used in the form of a salt. Examples of such salts include salts with inorganic bases, salts with inorganic acids, and salts with organic acids, and it is preferable to select a salt that is acceptable for use in medicines, foods, beverages, etc. Examples of salts with inorganic bases include alkali metal salts such as sodium, potassium, and lithium, alkaline earth metal salts such as calcium and magnesium, and ammonium salts. Examples of salts with inorganic acids include salts with hydrohalic acids (hydrochloric acid, hydrobromic acid, hydroiodic acid, etc.), sulfuric acid, nitric acid, phosphoric acid, etc. Examples of salts with organic acids include salts with formic acid, acetic acid, propionic acid, oxalic acid, succinic acid, maleic acid, fumaric acid, and citric acid.

[0052] In one embodiment of the present invention, the isothiocyanate derivative of the present invention may be prepared as is as described above, or may be concentrated, dried, and then dissolved again in water or an organic solvent, or may be purified by decolorization, deodorization, desalting, or fractionation by column chromatography using ion exchange resins. For storage, the isothiocyanate derivative may be freeze-dried after purification and dissolved in a solvent when used. In one embodiment of the present invention, the isothiocyanate derivative of the present invention may be prepared as described above and isolated, or may be purified as described above and used, or may be modified appropriately according to the purpose.

[0053] The isothiocyanate derivative of the present invention can be used as it is, or can be incorporated into an aqueous carrier such as water or a lower alcohol, or into a base such as an emulsion, gel, or cream, or can be powdered or granulated, or can be encapsulated in a vesicle such as a liposome, or a microcapsule, etc.

[0054] The isothiocyanate derivatives of the present invention may contain additives such as pharma- ceutically acceptable excipients, binders, lubricants, surfactants, disintegrants, coating agents, thickeners, pH adjusters, antioxidants, preservatives, colorants, etc. In other embodiments, the isothiocyanate derivatives of the present invention may further contain other active ingredients.

[0055] The dosage form (or intake form as food) of the isothiocyanate derivative of the present invention may be oral administration, or parenteral administration (ingestion) such as drip infusion, injection (intravenous administration), or external use, and is not particularly limited. When administered orally, general pharmaceutical dosage forms such as granules, fine granules, powders, coated tablets, tablets, powders, (micro)capsules, chewable tablets, syrups, juices, liquids, suspensions, emulsions, and other preparations that extend the release of active substances can be adopted. Furthermore, it can be formulated using various carriers or bases and additives, and administered in the form of sugar-coated tablets or tablets containing film coating agents, pills, capsules, ampoules, syrups, suspensions, emulsions, elixirs, drops, lozenges, chewable tablets, powders, granules, and other forms. When administered orally, it can be administered before, after, or between meals.

[0056] When administering parenterally, for example, a solution containing the active ingredient can be sprayed as a nasal drop or administered as an injection.Alternatively, when the isothiocyanate derivative of the present invention is used as a skin external preparation, the active ingredient can be dispersed in various bases and formulated by a conventional method, and examples of such bases include petrolatum, liquid paraffin, higher fatty acid esters such as isopropyl myristate and octyldodecyl myristate, higher alcohols such as squalane, lanolin, and cetanol, oily bases such as silicone oils and animal and vegetable oils, lower alcohols such as ethanol, polyhydric alcohols such as polyethylene glycol and propylene glycol, emulsifiers or emulsion stabilizers such as α-monoglyceryl ether, lecithin, sorbitan fatty acid esters, dextrin fatty acid esters, fatty acid monoglycerides, fatty acid metal salts, and magnesium sulfate, various medicinal agents such as fragrances, preservatives, pigments, thickeners, antioxidants, ultraviolet protection agents, wound healing agents, anti-inflammatory agents, and moisturizing agents, and water. Furthermore, the isothiocyanate derivative of the present invention can be added to external preparations as an antioxidant for the skin, and can be provided as external medicines, quasi-drugs, cosmetics, and the like.

[0057] The isothiocyanate derivative of the present invention is useful for application to humans, non-human animals (e.g., non-human mammals (livestock and pets such as pigs, cows, horses, and dogs), birds (poultry and pets such as turkeys and chickens), etc.), etc. It can also be added to veterinary medicines and animal feed.

[0058] The food to which the isothiocyanate derivative of the present invention is added is not particularly limited, but examples include those in which the active ingredient is added to food such as powdered drinks, beverages, confectionery, etc. The food may be food in general, and includes general foods including health foods, foods for specified health uses, foods with nutritional functions, and supplements.

[0059] In this specification, "or" is used when "at least one or more" of the items listed in the sentence can be employed. The same applies to "alternative." In this specification, when "within the range of" two values ​​is specified, the range includes the two values ​​themselves.

[0060] All references cited herein, including scientific literature, patents, patent applications, and the like, are hereby incorporated by reference in their entirety to the same extent as if each was specifically set forth.

[0061] The present disclosure has been described above by showing preferred embodiments for ease of understanding. The present disclosure will be described below based on examples, but the above description and the following examples are provided for illustrative purposes only and are not provided for the purpose of limiting the present disclosure. Therefore, the scope of the present disclosure is not limited to the embodiments or examples specifically described in this specification, but is limited only by the claims. In addition, unless otherwise specified, "%" means "% by weight". EXAMPLES

[0062] Example 1: Preparation of powdered ITCs 100 g of unsterilized dried moringa leaves were placed in a container, and 20 g of yeast extract (Mitsubishi Corporation Life Sciences Co., Ltd.) was added so that the amount was 0.2 times the weight of the moringa leaves. Furthermore, 1500 g of distilled water was added so that the amount was 15 times the weight of the moringa leaves. (extraction) The resulting mixture was stirred at room temperature for 60 minutes with constant stirring. (filtration) Then, 12 g of Radiolite #100 was added as a filter aid so that the weight ratio was 10% based on the weight of the raw materials, moringa leaves and yeast extract, and filtration was performed. (Input) Furthermore, 20 g of dextrin (Sandec #100, Sanwa Starch Industry Co., Ltd.) was added so that the weight ratio was 20% based on the weight of the moringa leaves. (sterilization) It was then heated to 90°C for 10 minutes. (Powdered) Finally, the mixture was powdered (spray dried: SD) using a spray dryer (SD-1010 type) to obtain stabilized ITCs (GSH-ITCs).

[0063] Example 2: Selection of stabilizers Since ITCs bind to cysteine ​​groups, cysteine, oxidized glutathione (GSSG), and reduced glutathione (GSH) were selected as candidates for stabilizers. Each stabilizer was added to a water extract of moringa leaves to prepare a mixed solution. With the aim of industrial application, the mixed solution was heated at 100°C for 10 minutes. The remaining rate was calculated from the total amount of ITCs before and after heating, and stabilizers were selected.

[0064] Moringa leaves were extracted with GSH, the solution was sterilized by heating, and dextrin was added to make powder (GSH-ITCs).GSH-ITCs were stored at 40℃ and 75% humidity for 45 days, and the total ITCs amount was measured before and after storage.

[0065] The results of the stabilizer selection test are shown in Figure 2. Comparing the three stabilizers, GSH showed the highest retention rate of ITCs. On the other hand, when GSSG was used, no binding to ITCs occurred. This is thought to be because GSSG cannot bind to ITCs because two GSHs are linked by disulfide bonds.

[0066] (Example 3: Examination of the amount of yeast extract added) 20 g of unsterilized dried Moringa leaves were placed in a container, and 0, 0.5, 1, 2, 4, or 6 g of yeast extract was added to the Moringa leaves so that the amount was 0, 0.025, 0.05, 0.1, 0.2, or 0.3 times the amount by weight of the Moringa leaves, respectively. In addition, 300 g of distilled water was added to the Moringa leaves so that the amount was 15 times the amount by weight of the Moringa leaves. (extraction) The resulting mixture was stirred at room temperature for 15 minutes with constant stirring. (Powderization) After extraction, the mixture was filtered and finally powdered using a freeze dryer (freeze drying: FD). The resulting composition was dissolved in water and heated at 100°C for 10 minutes to measure the residual isothiocyanate rate. The relationship between the amount of yeast extract added and the residual rate is summarized in Figure 3.

[0067] (Example 4: Study of extraction temperature) 20 g of unsterilized dried moringa leaves were placed in a container, and 4 g of yeast extract was added so that the amount was 0.2 times the weight of the moringa leaves. Furthermore, 300 g of distilled water was added so that the amount was 15 times the weight of the moringa leaves. (extraction) The resulting mixture was stirred for 15 minutes at low temperature (water temperature 5.7°C), room temperature (water temperature 21.7°C) or heated (water temperature 35.9°C) with constant stirring. (Powderization) After extraction, the mixture was filtered and finally powdered using a freeze dryer (freeze drying: FD), and the amount of isothiocyanate in the resulting composition was measured. The relationship between the extraction temperature and the amount of isothiocyanate is summarized in Figure 4. Isothiocyanates could be extracted at any extraction temperature.

[0068] (Example 5: Study of extraction time) 20g or 40g of unsterilized dried Moringa leaves were placed in a container, and 4g or 8g of yeast extract was added so that the amount was 0.2 times the weight of the Moringa leaves. Furthermore, 300g or 600g of distilled water was added so that the amount was 15 times the weight of the Moringa leaves. (extraction) The resulting mixture was stirred at room temperature for 15, 60 or 180 minutes with constant stirring. (filtration) Thereafter, Radiolite #100 was added as a filter aid at 2.4 g or 4.8 g so as to be 10% by weight based on the raw material moringa leaves and yeast extract, and filtration was performed. (Input) Furthermore, cyclodextrin γ was added at 4 g or 8 g so that the amount was 0.2 times the weight of the moringa leaves. (Powdered) Finally, the mixture was powdered using a spray dryer (spray drying: SD), and the amount of isothiocyanate in the obtained composition was measured. The relationship between the extraction time and the amount of isothiocyanate is summarized in Figure 5. Isothiocyanates could be extracted at any extraction time.

[0069] (Example 6: Examination of filter aids) 20 g of unsterilized dried moringa leaves were placed in a container, and 4 g of yeast extract was added so that the amount was 0.2 times the weight of the moringa leaves. Furthermore, 300 g of distilled water was added so that the amount was 15 times the weight of the moringa leaves. (extraction) The resulting mixture was stirred at room temperature for 15 minutes with constant stirring. (filtration) Thereafter, filtration was performed without adding any filter aid, or with the addition of 2.4 g of Radiolite #100 or Radiolite #700 as a filter aid, so that the weight of the moringa leaves and yeast extract used as raw materials was 10%. (Powdered) Finally, the mixture was powdered using a freeze dryer (freeze drying: FD), and the amount of isothiocyanate in the resulting powder was measured. The relationship between the amount of filter aid and the amount of isothiocyanate is summarized in Figure 6. None of the filter aids affected the amount of isothiocyanate extracted.

[0070] (Example 7: Study of dextrin) 50g or 40g of unsterilized dried Moringa leaves were placed in a container, and 10g or 8g of yeast extract was added so that the amount was 0.2 times the weight of the Moringa leaves. Furthermore, 750g or 600g of distilled water was added so that the amount was 15 times the weight of the Moringa leaves. (extraction) The resulting mixture was stirred at room temperature for 15 minutes (180 minutes for dextrin) with constant stirring. (filtration) Then, 6g or 4.8g of Radiolite #100 was added as a filter aid to the raw materials, the chopped moringa leaves and yeast extract, so that the weight ratio was 10%, and filtration was performed. (Input) Further, the dextrins shown in the table below were added to the moringa leaves in amounts per weight shown in the table below. (Powdered) Finally, the mixture was powdered using a freeze dryer (freeze drying: FD), and the resulting powder was stored at 40°C and 75% humidity for 25 days (45 days for dextrin), and the storage stability was confirmed and summarized in Table 1. None of the dextrins caking occurred when added in an amount of 0.2 or more times the original amount. [Table 1]

[0071] (Example 8: Study of sterilization mode) 100 g of unsterilized dried moringa leaves were placed in a container, and 20 g of yeast extract was added so that the amount was 0.2 times the weight of the moringa leaves. Furthermore, 1500 g of distilled water was added so that the amount was 15 times the weight of the moringa leaves. (extraction) The resulting mixture was stirred at room temperature for 15 minutes with constant stirring. (filtration) Then, 12 g of Radiolite #100 was added as a filter aid so that the weight ratio was 10% based on the weight of the raw materials, moringa leaves and yeast extract, and filtration was performed. (Input) Furthermore, 20 g of cyclodextrin γ was added so that the weight ratio was 20% relative to the weight of the moringa leaves. (sterilization) The sample was then either not heated, heated at 90°C for 10 minutes, or heated at 121°C for 2 minutes. (Powderization) Finally, the mixture was pulverized using a freeze dryer, and the amount of isothiocyanate in the resulting powder was measured. The relationship between the state of sterilization and the amount of isothiocyanate is summarized in Figure 7. It was confirmed that heat sterilization for 10 minutes after reaching at least 90°C, or heat sterilization for 2 minutes at 121°C, resulted in a negative result for E. coli and a general viable cell count of 3000 CFU or less. In addition, heat sterilization at 90°C for 10 minutes was able to suppress the reduction in the amount of isothiocyanate to about 70% compared to no sterilization.

[0072] (Example 9: Examination of powderization) The same operations as in Example 1 were carried out up to the (sterilization) step prior to the (powdering) step. (Powderization) Next, the mixture was powdered using a freeze dryer (freeze dryer: FD) or a spray dryer (spray dryer: SD), and the amount of isothiocyanate in the resulting powder was measured. The relationship between the powdering process and the amount of isothiocyanate is summarized in Figure 8. There was no change in the amount of isothiocyanate with either powdering method.

[0073] Example 10: Anti-obesity effect The anti-obesity effect of GSH-ITCs was confirmed. C57BL / 6J mice (male, 5 weeks old) were given 1. a standard diet (STD), 2. a high-fat diet (HFD), 3. an HFD + stabilized ITCs (GSH-ITCs), or 4. an HFD + Moringa-derived ITCs ad libitum for 4 weeks, and their body weight was measured every week (3, 4: total ITCs (including isothiocyanate derivatives) approximately 0.37 mg / kg BW).

[0074] The results are shown in Figure 9. GSH-ITCs significantly suppressed weight gain due to HFD intake (p<0.05), with an effect similar to that of Moringa-derived ITCs. Moringa-derived ITCs maintained their original anti-obesity effect even when bound to GSH, while increasing their stability against heat sterilization, and are therefore expected to contribute to healthy longevity and be applied in industry.

[0075] (Note) While the present invention has been illustrated by way of preferred embodiments thereof, it is understood that the scope of the present invention should be interpreted only by the claims. It is understood that the patents, patent applications, and other documents cited in this specification are incorporated by reference into this specification in the same manner as if the contents themselves were specifically set forth herein. [Industrial Applicability]

[0076] INDUSTRIAL APPLICABILITY The present invention can provide a stable isothiocyanate having useful physiological functions, and is therefore useful in the functional material industry for preventing diseases and supporting healthy longevity.

Claims

1. formula 【Chemistry 11】 or a pharma- ceutically acceptable salt thereof, wherein R １ , R ２ and R ３ is an acetyl group or H, R ４ is OH or an amino acid having 1 to 3 residues, R ５ is H or 1-3 amino acid residues; The compound, or a pharma- ceutically acceptable salt thereof.

2. R １ , R ２ and R ３ is H or R １ is an acetyl group, and R ２ and R ３ is H or R ２ is an acetyl group, and R １ and R ３ is H, or R ３ is an acetyl group, and R １ and R ２ is H, R ４ but, 【Chemistry 12】 (where A represents the attachment point) and R ５ but, 【Chemistry 13】 (where A represents the attachment point) That is, 2. The compound of claim 1, or a pharma- ceutically acceptable salt thereof.

3. R １ , R ２ and R ３ is H or R １ is an acetyl group, and R ２ and R ３ is H or R ２ is an acetyl group, and R １ and R ３ is H, or R ３ is an acetyl group, and R １ and R ２ is H, R ４ is OH, R ５ is H, 2. The compound of claim 1, or a pharma- ceutically acceptable salt thereof.

4. R １ , R ２ and R ３ is H or R １ is an acetyl group, and R ２ and R ３ is H or R ２ is an acetyl group, and R １ and R ３ is H, or R ３ is an acetyl group, and R １ and R ２ is H, R ４ is OH, R ５ but 【Chemistry 14】 (where A represents the attachment point) 2. The compound of claim 1, wherein:

5. 5. A method for producing the compound of any one of claims 1 to 4 or a pharma- ceutically acceptable salt thereof, comprising reacting an extract of a plant of the Moringa family and a yeast extract containing cysteine ​​peptides in an aqueous medium.

6. The method of claim 5 , wherein the cysteine ​​peptide is glutathione.

7. 6. The method of claim 5, wherein the extract of a plant from the Moringa family is an extract of Moringa oleifera.

8. The method of claim 5 , wherein the plant extract of the Moringaceae family contains moringin and / or an acetylated form of moringin.

9. 6. The method according to claim 5, wherein the amount of the yeast extract added is about 10% by weight or more relative to the amount of the extract of a plant of the Moringa family added.

10. The method according to claim 5 , wherein the yeast extract is a yeast extract derived from Torula yeast or a yeast extract derived from Saccharomyces cerevisiae.

11. 6. The method of claim 5 further comprising the step of adding dextrin.

12. The method according to claim 11, wherein the amount of dextrin added is about 20% by weight or more relative to the amount of the plant extract of the Moringa family added.

13. The method according to claim 5, further comprising the step of heat sterilizing the aqueous solution obtained by the reacting step at about 65°C or higher.

14. A composition comprising the compound according to any one of claims 1 to 4 or a pharma- ceutical acceptable salt thereof, and having anti-obesity, weight gain suppression, lipid metabolism improvement, liver function improvement, and / or fatty liver prevention / improvement effects.

Images