Prodrugs of ester derivatives of inotodiol

Abstract

Regarding a prodrug of an ester derivative of inotoxin and its use, the prodrug not only has increased solubility in inorganic and organic solvents but also has improved stability, and can be used to treat various inflammatory diseases, including inflammatory or allergic diseases.

Description

[Technical field]

[0001] This relates to prodrugs of ester derivatives of inotodiol. [Background technology]

[0002] A prodrug is a drug that is initially inactive, but becomes active through a metabolic process after being administered to the body, and exerts its effects. A prodrug combines a therapeutic agent (A), which is broken down immediately after being administered into the body, with another substance (B) and delivers it to target cells or tissues. After that, B is excreted through the body's metabolic process, leaving only component A, which allows the treatment to be carried out.

[0003] On the other hand, inotodiol is a compound isolated from the alcohol extract of chaga and has the characteristics of an unsaturated diol. Inotodiol is not water-soluble, has a relatively low solubility in organic solvents, is unstable and easily precipitates, but has a low absorption rate in the body when administered orally. In addition, the fraction that does not precipitate when administered orally is easily absorbed in the small intestine, so it cannot reach the terminal small intestine or large intestine, and the precipitated inotodiol does not show biological activity and is excreted from the body.

[0004] Therefore, in order to overcome the above problems, it is necessary to develop a prodrug of inotodiol having improved physicochemical, physiological and pharmaceutical properties. Summary of the Invention [Problem to be solved by the invention]

[0005] One aspect is to provide a prodrug of an ester derivative of inotodiol or a pharma- ceutically acceptable salt thereof.

[0006] Another aspect is to provide a pharmaceutical composition for preventing or treating an inflammatory disease or an allergic disease, comprising the prodrug or a pharma- ceutically acceptable salt thereof as an active ingredient.

[0007] Another aspect is to provide a health functional food for preventing or ameliorating inflammatory diseases or allergic diseases, which contains the prodrug or a pharma- ceutically acceptable salt thereof as an active ingredient.

[0008] Another aspect is to provide a method of preventing or treating an inflammatory or allergic disease comprising administering to an individual in need thereof an effective amount of said prodrug or a pharma- ceutically acceptable salt thereof. [Means for solving the problem]

[0009] One embodiment provides prodrugs of ester derivatives of inotodiol.

[0010] One embodiment provides an ester derivative compound of inotodiol formed by ester condensation reaction between a hydroxyl group of an inotodiol compound of the following chemical formula 1 and a carboxyl group of a fatty acid, glucuronic acid, alkenyl succinic anhydride, or phenolic acid, or a pharma- ceutically acceptable salt thereof. [ka]

[0011] One embodiment provides a compound represented by the following chemical formula 2, or a pharma- ceutically acceptable salt thereof: [ka] In the above formula, R 1 and R 2 are each independently OH or -OC(O)-R 3 and R 3 is a linear or branched alkyl, alkenyl, or alkynyl chain having 1 to 30, 4 to 30, or 6 to 30 carbon atoms; R 1 and R 2 At least one of them is -OC(O)-R 3 It is.

[0012] In one embodiment, the R 3 is CH3(CH2) a - or CH3(CH2) b (CH=CH[CH2]) c (CH2) d -, wherein a is an integer from 8 to 24, b is an integer from 1 to 5, c is an integer from 1 to 6, and d is an integer from 3 to 7.

[0013] In the present specification, the term "prodrug" may refer to a drug that may be biologically inactive by itself, but exhibits effective pharmacological effects after chemical / biochemical structural modification during its residence time in the body. That is, a drug that is useful but has unsuitable properties in terms of side effects, stability, solubility, absorbability, duration of action, etc., is chemically modified to enable clinical use. In one embodiment, the inotodiol derivative is a compound formed through an ester bond between inotodiol and a fatty acid, and not only has increased solubility in inorganic solvents and / or organic solvents and improved stability, but is also easily decomposed into inotodiol and a fatty acid in the small intestine, making it easily absorbed in the small intestine. For example, the inotodiol derivative may be formed through an ester bond between inotodiol and succinic acid. Specifically, the inotodiol derivative can be synthesized by condensation reaction of alkenyl succinic anhydride and inotodiol under p-toluenesulfonic acid catalyst, and succinic acid ester bond can be induced to all or at least one hydroxyl group of inotodiol by adding alkyl succinic anhydride in excess of inotodiol. After completion of the reaction, p-toluenesulfonic acid is neutralized and removed with sodium bicarbonate. The inotodiol derivative compound synthesized by the above method is R of Chemical Formula 2. 1 and R 2Both or at least one of the inotodiol derivatives form a succinic acid ester bond, improving water solubility, and the ester bond is easily decomposed to facilitate absorption in the small intestine. In addition, the inotodiol derivative according to another embodiment not only increases the water solubility of inotodiol through the ester bond between inotodiol and a fatty acid, but also reduces hydrolysis in the small intestine and absorption in the body, increasing the possibility that the inotodiol derivative reaches the large intestine, and when the inotodiol derivative is decomposed by various microorganisms in the large intestine, inotodiol can directly act pharmacologically. For example, the inotodiol derivative may be formed through an ester bond between inotodiol and a phenolic acid. Specifically, the inotodiol derivative is expected to have various physiological activities of inotodiol and phenolic acid by hydrolyzing the ester bond in the large intestine to release inotodiol and phenolic acid. Therefore, the prodrug according to one embodiment can not only reduce the dosage of inotodiol, but also reduce side effects that may occur when it is taken at a high concentration.

[0014] The inotodiol, as the main physiologically active component of chaga (Inonotus obliquus), can be chemically synthesized according to conventional methods, can be produced from a pharma- ceutically acceptable salt, or can be isolated and purified from an extract of chaga.

[0015] As used herein, the term "inotodiol" refers to the aryl ester having the IUPAC name (3S,5R,10S,13R,14R,17R)-17-[(2S,3R)-3-hydroxy-6-methylhept-5-en-2-yl]-4,4,10,13,14-pentamethyl-2,3,5,6,7,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthrene. nthren-3-ol((3S,5R,10S,13R,14R,17R)-17-[(2S,3R)-3-hydroxy-6-methylhept-5-en-2-yl]-4,4,10, 13,14-pentamethyl-2,3,5,6,7,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-3-ol).

[0016] The inotodiol derivative can be synthesized by chemical / biological methods. Specifically, the inotodiol derivative can be synthesized by esterification of inotodiol with a fatty acid. The fatty acid can be C 10 ~C 30 It can be unsaturated or saturated fatty acid, C1-C 10The unsaturated fatty acid may be selected from the group consisting of carboxylic acids and phenolic acids. The unsaturated fatty acid may be, for example, myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, etc. The saturated fatty acid may be, for example, caprylic acid, capric acid, lauric acid, palmitic acid, stearic acid, arachic acid, behenic acid, lignoceric acid, cerotic acid, and myristic acid, etc. The saturated carboxylic acid may be, for example, formic acid, acetic acid, propionic acid, butyric acid, succinic acid, etc. The phenolic acid may be, for example, p-coumaric acid, cinnamic acid, ferulic acid, 3,4-dihydroxy benzoic acid, p-hydroxy benzoic acid, vanillic acid, caffeic acid, syringic acid, sinapinic acids, etc. In one embodiment, the inotodiol derivative is a mixture of inotodiol and an unsaturated fatty acid, a saturated fatty acid, and a C1 to C 10The esterification reaction may be carried out at 55 to 65° C. for 48 to 96 hours. If the esterification reaction conditions are below or above the range, it is difficult to completely dissolve the fatty acid, which is the reaction substrate, and the reaction hardly occurs, or the enzyme activity is reduced, which reduces the esterification reaction efficiency, and the final product, the inotodiol derivative, cannot be synthesized in high yield. After the esterification reaction is completed, the purity of the ester contained in the product can be further increased by various known distillation or purification methods.

[0017] In addition, the inotodiol derivative can be synthesized by adding a biological enzyme to the mixture of the inotodiol and fatty acid. The biological enzyme may be, for example, Candida antarctica Lipase B (CalB), which is an enzyme suitable for producing the inotodiol derivative with high efficiency. The Candida antarctica Lipase B may be an immobilized enzyme rather than a general enzyme in order to increase the yield of the final product, the inotodiol derivative. The immobilized enzyme may be a commercially available product or may be prepared by a conventional method. In addition, the inotodiol derivative can be synthesized by adding 400 to 500 parts by weight of the Candida antarctica Lipase B based on 100 parts by weight of inotodiol. In this case, if the content of Candida antarctica Lipase B is less than the above range, there is a problem that the esterification reaction does not occur sufficiently and the efficiency of producing the ester is low, and if the content exceeds the above range, there is a problem that the final product, the inotodiol derivative, cannot be synthesized in high yield and is not economical.

[0018] One embodiment provides a composition for preventing or treating an allergic disease, which comprises the prodrug as an active ingredient.

[0019] The term "inflammatory disease" as used herein includes acute and / or chronic inflammatory diseases. The inflammatory disease may be selected from the group consisting of, for example, dermatitis, conjunctivitis, periodontitis, rhinitis, otitis media, pharyngitis, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, shoulder periarthritis, tendonitis, tendonitis, peritendinitis, myositis, hepatitis, cystitis, nephritis, Sjogren's syndrome, arteriosclerosis, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), multiple sclerosis, acute inflammatory disease, and chronic inflammatory disease.

[0020] The term "allergy" as used herein refers to a phenomenon in which an acute reaction occurs in vivo through an antigen-antibody reaction via contact with an allergen, which is an allergy-inducing substance. The allergy may be an allergic disease induced by, for example, pollen, dust mites, drugs, plant fibers, bacteria, food, dyes, chemicals, food, etc.

[0021] The allergic diseases are diseases caused by allergies, such as urticaria, anaphylaxis, allergic rhinitis, bronchial asthma, atopic dermatitis, drug rash, drug allergy, serum sickness, etc., and can refer to various types of diseases depending on the type of allergen and the tissue that causes the allergic reaction.

[0022] The pharmaceutical composition for preventing or treating an allergic disease according to one embodiment may be formulated and used in the form of oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, topical preparations, suppositories, and sterile injectable solutions, each according to a conventional method, and may contain suitable carriers, excipients, or diluents that are conventionally used in the manufacture of pharmaceutical compositions for formulation.

[0023] The carriers or excipients and diluents may include various compounds or mixtures including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like.

[0024] When the pharmaceutical preparation is prepared, it can be produced using a diluent or excipient that is commonly used, such as a filler, weighting agent, binder, wetting agent, disintegrant, or surfactant.

[0025] A solid preparation for oral administration can be prepared by mixing the inotodiol, a prodrug thereof, or a pharma- ceutically acceptable salt thereof with at least one excipient, such as starch, calcium borate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used.

[0026] Oral liquid preparations include suspensions, liquid preparations, emulsions, syrups, etc., and in addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, flavorings, preservatives, etc. may be contained.

[0027] Preparations for parenteral administration include sterile aqueous solutions, water-insoluble agents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can include witepsol, macrogol, tween 20, cacao butter, laurin butter, and glycerin gelatin.

[0028] The preferred dosage of the pharmaceutical composition for preventing or treating inflammatory or allergic diseases according to one embodiment varies depending on the patient's condition, body weight, degree of disease, drug form, administration route and period, but can be appropriately selected by those skilled in the art. However, for a favorable effect, it can be administered at 0.0001 to 2,000 mg / kg per day, preferably 0.001 to 2,000 mg / kg per day. The administration may be once a day or in several divided doses. However, the scope of the present invention is not limited by the above dosage.

[0029] The pharmaceutical composition for preventing or treating an inflammatory or allergic disease according to one embodiment can be administered to mammals such as rats, mice, livestock, humans, etc., by various routes. All of the methods of administration can be, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura or intracerebroventricular injection.

[0030] Another aspect provides a health functional food composition or cosmetic composition for preventing or improving inflammatory diseases or allergic diseases, comprising the compound or a pharma- ceutically acceptable salt thereof as an active ingredient.

[0031] The specific details of the above compounds are as described above.

[0032] In the health functional food for preventing or improving inflammatory disease or allergic disease according to one embodiment, when the compound is used as an additive of the health functional food, it can be added as it is or can be used in combination with other foods or food ingredients, and can be used appropriately according to conventional methods. The amount of the active ingredient mixed can be appropriately determined according to each purpose of use, such as prevention, health or treatment.

[0033] Health functional foods can be formulated not only in the form of powders, granules, pills, tablets, or capsules, but also in the form of general foods or beverages.

[0034] The type of food is not particularly limited, and examples of foods to which the substance can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, drinking water, tea, energy drinks, alcoholic beverages, and vitamin complexes, and may include all foods in the usual sense.

[0035] In general, when producing food or beverage, the compound can be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, per 100 parts by weight of raw material. However, in the case of long-term intake for the purpose of health and hygiene or health regulation, the amount may be less than the above range, and since there is no safety problem in using fractions from natural products, the amount may be more than the above range.

[0036] Among the health functional foods according to one embodiment, the beverage may contain various flavoring agents or natural carbohydrates as additional ingredients, as in the case of ordinary beverages. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as thaumatin and stevia extract, and artificial sweeteners such as saccharin and aspartame may be used. The proportion of natural carbohydrates may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 mL of the beverage according to the present invention.

[0037] In addition to the above, the health functional food according to one embodiment may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonation agents used in carbonated drinks. Furthermore, the sleep-improving composition of the present invention may contain fruit pulp for the production of natural fruit juices, fruit juice drinks, and vegetable drinks. These ingredients may be used independently or in mixture. The proportion of such additives is not limited, but is generally selected from the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health functional food of the present invention. Effect of the Invention

[0038] The prodrug according to one embodiment not only has increased solubility in inorganic and organic solvents but also has improved stability, and can be used to treat various inflammatory diseases, including inflammatory or allergic diseases. [Brief description of the drawings]

[0039] [Figure 1] FIG. 1 shows the HPLC-ELSD results of the oleic acid prodrug of inotodiol. [Diagram 2] FIG. 2 shows the HPLC-ELSD results of the linoleic acid prodrug of inotodiol. [Diagram 3] FIG. 3 shows the HPLC-ELSD results of the palmitic acid prodrug of inotodiol. [Figure 4] FIG. 4 shows the HPLC-ELSD results of the succinic acid prodrug of inotodiol. [Figure 5A] Figure 5 shows the LC-MS / MS results of the oleic acid prodrug of inotodiol. Figure 5A shows the molecular weight of the oleic acid ester of inotodiol. [Figure 5B] FIG. 5B is a diagram confirming the fragment pattern of the oleate ester of inotodiol. [Figure 6A] Figure 6 shows the results of LC-MS / MS of the succinic acid prodrug of inotodiol. Figure 6A shows the molecular weight of the succinic acid ester of inotodiol. [Figure 6B] FIG. 6B is a diagram confirming the fragmentation pattern of the succinate ester of inotodiol. [Figure 7A] Figure 7 shows the results of LC-MS / MS of the linoleic acid prodrug of inotodiol. Figure 7A shows the molecular weight of the linoleic acid ester of inotodiol. [Figure 7B] FIG. 7B is a diagram confirming the fragment pattern of the linoleic acid ester of inotodiol. [Figure 8A]Figure 8 shows the LC-MS / MS results of the palmitic acid prodrug of inotodiol. Figure 8A shows the molecular weight of the palmitic acid ester of inotodiol. [Figure 8B] FIG. 8B is a diagram confirming the fragment pattern of the palmitate ester of inotodiol. [Figure 9A] Figure 9 shows the 1H-NMR results of the oleic acid prodrug of inotodiol. Figure 9A shows the 1H-NMR of inotodiol. [Figure 9B] FIG. 9B is a diagram confirming 1H-NMR of 22-O-oleyl-inothion diol; [Figure 9C] FIG. 9C shows the 1H-NMR of 3-O-oleyl-inothionyldiol. [Figure 10A] FIG. 10 shows the results of 13C-NMR of the oleic acid prodrug of inotodiol, and FIG. 10A shows the 13C-NMR of inotodiol. [Figure 10B] FIG. 10B is a diagram confirming 13C-NMR of 22-O-oleyl-inothion diol; [Figure 10C] FIG. 10C shows the 13C-NMR of 3-O-oleyl-inothionyldiol. [Figure 11A] FIG. 11 shows the results of two-dimensional heteronuclear multicomponent correlation (HMBC) of the oleic acid prodrug of inotodiol. FIG. 11A shows the two-dimensional 1H-13C HMBC of 22-O-oleyl-inotodiol. [Figure 11B] FIG. 11B shows the two-dimensional 1H-13C HMBC of 3-O-oleyl-inotodiol. [Figure 12A] FIG. 12 shows the DSC results of an oleic acid prodrug of inotodiol; FIG. 12A shows the DSC results of inotodiol; [Figure 12B] FIG. 12B shows the DSC results of the oleate ester of inotodiol. [Figure 13A]Figure 13 shows the DSC results of the succinic acid prodrug of inotodiol. Figure 13A shows the DSC results of inotodiol. [Figure 13B] FIG. 13B shows the DSC results of inotodiol succinate. [Figure 14A] FIG. 14 shows the FT-IR results of the oleic acid prodrug of inotodiol, and FIG. 14A shows the FT-IR results of inotodiol. [Figure 14B] FIG. 14B shows the FT-IR results of inotodiol oleate. [Figure 15A] Figure 15 shows the FT-IR results of the succinic acid prodrug of inotodiol. Figure 15A shows the FT-IR results of inotodiol. [Figure 15B] FIG. 15B shows the FT-IR result of inotodiol succinate. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0040] In the following, preferred examples are presented to aid in understanding the present invention. However, the following examples are provided to facilitate understanding of the present invention, and are not intended to limit the scope of the present invention. EXAMPLES

[0041] Example 1 Measurement of water solubility of inotodiol To measure the water solubility of inotodiol, the following procedure was carried out. First, 50 mg of inotodiol sample was mixed with 1 ml of organic solvent. Then, it was vortexed and placed in a water bath sonicator for 15 minutes. After leaving it at 25°C for 1 hour, it was centrifuged at 13,000 rpm for 5 minutes. Then, the supernatant was collected and subjected to HPLC-UV-VIS analysis, and the results are shown in Table 1 below. [Table 1]

[0042] Example 2 Enzymatic synthesis and purification of fatty acid esters of inotodiol To prepare esters of inotodiol, various acyl donors including oleic acid, linoleic acid, palmitic acid and succinic acid were esterified with inotodiol (molar ratio inotodiol / acid = 1:4) by lipase reaction. First, 0.1 mmol of inotodiol and 0.4 mmol of acid were prepared in a 50 mL glass bottle with a screw cap and dissolved in 5 mL of t-butanol for 30 min at 60 °C using a magnetic stirrer (400 rpm). Then, 0.5 mL of enzyme solution (Candida antarctica lipase A) was added to the mixture, and the reaction was carried out at 60 °C for 72 h with stirring at 400 rpm during incubation. Ethanol (10 mL) was added to the reaction solution to stop the reaction and kept at 60 °C for 30 min to precipitate the lipase. The solution was then filtered to remove the enzyme. Finally, the supernatant was collected and concentrated to a small volume under vacuum using a rotary evaporator (Eyela, Tokyo, Japan). Next, the concentrated mixture (1 mL) was loaded onto a glass column (15x250 mm) filled with ODS-AQ C18 (YMCKOREA, Seongnam, Korea) via a preparative HPLC system (LC-Forte / R, YMCKOREA) at a flow rate of 10 mL / min to purify the esterified products. For the purification of inotodiol oleate ester, inotodiol linoleate ester, and inotodiol palmitate ester, the mobile phase was distilled water (solvent A) and methanol:ethanol = 1:1 (solvent B), and eluted with the following gradient elution method: (0-10 min, 90-95% B), (10-25 min, 95-100% B), (25-80 min, 100% B). For the purification of inotodiol succinate ester, the following gradient elution was used: (0-10 min, 0-10% B), (10-30 min, 10-50% B), (30-80 min, 100% B). The effluent was continuously monitored at a detection wavelength of 210 nm. The peak fractions of inotodiol ester were manually collected by observing the chromatogram.

[0043] Experimental Example 1: High-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) analysis HPLC analysis was performed on an Ultimate 3000 UHPLC system (Ultimate 3000, Dionex, Idstein, Germany) equipped with an RS diode array detector (Dionex) and a YMC-triart C18 column (4.6 mm × 250 mm, YMCKOREA, Sungnam, South Korea). Chromatographic separation was performed using a binary mobile phase gradient consisting of water (solvent A) and methanol (solvent B). For inotodiol oleate ester, inotodiol linoleate ester, and inotodiol palmitate ester, the solvent B gradient was 85% at 0 min, 92% at 7 min, 94% at 15 min, 100% at 20 min, 100% at 50 min, 85% at 50.1 min, and 85% at 55 min. For inotodiol succinate ester, the solvent B gradient was 0% at 0 min, 5% at 5 min, 50% at 12 min, 100% at 15 min, 100% at 30 min, 0% at 30.1 min, and 0% at 40 min. The injection volume was 10 μL and elution was monitored at 210 nm. The flow rate was set at 1 mL / min with a column temperature of 50 °C. ELSD analysis was performed on a Waters 1525 Binary HPLC system (Waters Corp.) equipped with an evaporative light scattering detector (Alltech2000, Deerfield, IL, USA) and a YMC-triart C18 column (4.6 mm × 250 mm, YMCKOREA, Sungnam, South Korea). The injection volume was 10 μL, and the eluent was monitored by ELSD with a drift tube temperature of 90 °C, the transport gas flow rate was 2.0 L / min, and the detector was set at 1. The flow rate was set at 1 mL / min with a column temperature of 50 °C. As a result, as shown in Figures 1 to 4, it was found that the synthesis of inotodiol prodrugs, i.e., inotodiol oleate ester, inotodiol linoleate ester, inotodiol palmitate ester, and inotodiol succinate ester, was carried out successfully. FIG. 1 shows the HPLC-ELSD results of the oleic acid prodrug of inotodiol. FIG. 2 shows the HPLC-ELSD results of the linoleic acid prodrug of inotodiol. FIG. 3 shows the HPLC-ELSD results of the palmitic acid prodrug of inotodiol. FIG. 4 shows the HPLC-ELSD results of the succinic acid prodrug of inotodiol.

[0044] Experimental Example 2 High-performance liquid chromatography tandem mass spectrometry (LC / MS) Esters of inotodiols were identified using an Agilent-1290 UPLC / 6470A and Agilent-1290 UPLC / 6430A QQQ system (Agilent Technologies Korea, Seoul, South Korea) equipped with an ESI (electrospray ionization) source. The mobile phase consisted of solution A (distilled water) and solution B (methanol) with 5 mM ammonium acetate and 0.1% formic acid in each solvent. Separation was performed on a column equipped with a Zorbax Eclipse Plus C18 column (2.1 × 50 mm, 3.5 μm) (Agilent Technologies Korea). The solution B gradient was 85% from 0 to 2 min, 90% from 7 min, 100% from 18 to 25 min, and 85% from 26 to 35 min. The injection volume was 2 μL, and the flow rate was set at 0.2 mL / min with a column temperature of 40 °C. The mass spectrometer was run in positive mode using the following parameters: Drying gas temperature: 270°C, drying gas flow rate: 10 L / min, spray pressure: 40 psi, sheath gas temperature: 300°C, sheath gas flow rate: 11 L / min, capillary voltage: 3500 V (positive), nozzle voltage: 500 V (positive). As a result, as shown in Figures 5 to 8, specific peaks were confirmed for each of the prodrugs of inotodiol: inotodiol oleate ester, inotodiol linoleate ester, inotodiol palmitate ester, and inotodiol succinate ester. FIG. 5A is a diagram confirming the molecular weight of the oleic acid ester of inotodiol, and FIG. 5B is a diagram confirming the fragment pattern of the oleic acid ester of inotodiol. FIG. 6A is a diagram confirming the molecular weight of the succinate ester of inotodiol, and FIG. 6B is a diagram confirming the fragment pattern of the succinate ester of inotodiol. FIG. 7A is a diagram confirming the molecular weight of linoleic acid ester of inotodiol, and FIG. 7B is a diagram confirming the fragment pattern of linoleic acid ester of inotodiol. FIG. 8A is a diagram confirming the molecular weight of the palmitate ester of inotodiol, and FIG. 8B is a diagram confirming the fragment pattern of the palmitate ester of inotodiol.

[0045] Experimental Example 3 Nuclear magnetic resonance (NMR) analysis For NMR analysis, the oleate ester of inotodiol (5 mg) was dissolved in 600 μL of CDCl3 and then transferred to an NMR tube (Eurisotop, Saint Aubin, France). 1 H NMR and 13 C NMR spectra were obtained on a Bruker Avance-III-600 (Bruker, Seongnam, South Korea) instrument at frequencies of 600.23 MHz and 150.93 MHz, respectively. Two-dimensional heteronuclear multibond correlation (HMBC) was used to identify the ester bond formation and chemical structure of the oleate ester of inotodiol. The two-dimensional HMBC NMR spectrum was obtained using 32 scans for 13 h. The data were analyzed using TopSpin TM They were processed using the educational version of 4.0 software (Bruker). As a result, the structure of the oleic acid ester of inotodiol was confirmed, as shown in Figs. FIG. 9A shows the structure of inotodiol. 1 FIG. 9B shows the H-NMR of 22-O-oleyl-inothionyl diol. 1 9C is a diagram showing the H-NMR of 3-O-oleyl-inothionyldiol. 11H-NMR confirmed. FIG. 10A shows the structure of inotodiol. 13 FIG. 10B shows the C-NMR of 22-O-oleyl-inothionyl diol. 13 FIG. 10C shows the C-NMR spectrum of 3-O-oleyl-inothionyl diol. 13 This is a diagram confirming C-NMR. FIG. 11A shows a two-dimensional representation of 22-O-oleyl-inothion diol. 1 H- 13 11C HMBC. FIG. 11B shows the 2D structure of 3-O-oleyl-inothionyldiol. 1 H- 13 C HMBC confirmed.

[0046] Experimental Example 4 Differential Scanning Calorimetry (DSC) Thermal analysis of inotodiol oleate and inotodiol succinate was performed on a DSC-1 instrument (Mettler Toledo, Greifensee, Switzerland). Approximately 2 mg of each dried sample was placed in a sealed aluminum pan. The pan was then heated from 25 °C to 400 °C at a rate of 10 °C / min under nitrogen atmosphere. An empty pan was used as a reference. As a result, as shown in FIG. 12 and FIG. 13, it was confirmed that the oleic acid ester of inotodiol and the succinic acid ester of inotodiol had lower melting points than inotodiol. FIG. 12A shows the DSC results of inotodiol, and FIG. 12B shows the DSC results of the oleate ester of inotodiol. FIG. 13A shows the DSC results of inotodiol, and FIG. 13B shows the DSC results of the succinate ester of inotodiol.

[0047] Experimental Example 5 Fourier transform infrared spectroscopy (FT-IR) FT-IR analysis was performed on the oleate ester of inotodiol and the succinate ester of inotodiol. Infrared spectra were recorded on a vacuum infrared spectrometer (VERTEX 80v) with a ratio of approximately 1 mg of sample to 100 mg of KBr (1:100). The spectra were measured from 4000 to 400 cm. -1 The average scan was obtained over a frequency range of 100 Hz to 100 Hz. As a result, as shown in Figs. 14 and 15, the structures of the oleic acid ester of inotodiol and the succinic acid ester of inotodiol were confirmed. FIG. 14A shows the FT-IR results of inotodiol, and FIG. 14B shows the FT-IR results of the oleate ester of inotodiol. FIG. 15A shows the FT-IR results of inotodiol, and FIG. 15B shows the FT-IR results of the succinate ester of inotodiol.

[0048] Experimental Example 6 Confirmation of the solubility of fatty acid esters of inotodiol To measure the solubility of inotodiol esters in water, 5 mg of sample was suspended in 1 mL of distilled water and then treated in an ultrasonic water bath for 30 min. The mixture was then treated in an ultrasonic water bath (Power sonic 410) for 30 min. The mixture was then incubated at 30 °C for 3 h. The supernatant obtained after centrifugation (20,000 x g, 10 min) was filtered through a 0.45 μm membrane filter from ADVANTEC (Tokyo, Japan) to remove undissolved crystals. The measurement of inotodiol succinate esters was performed with an Agilent-1290 UPLC / 6470A (Agilent Technologies Korea, Seoul, South Korea). The concentration of inotodiol succinate esters in the supernatants was measured according to an established standard curve (0–5 mg / mL, R 2 = 0.9998). As a result, as shown in Tables 2 and 3 below, it was confirmed that the solubility of the fatty acid esters of inotodiol, which are the precursors, was higher than that of inotodiol. [Table 2] [Table 3]

[0049] The above description of the present invention is for illustrative purposes only, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical idea or essential features of the present invention. Therefore, it should be understood that the above-described embodiments are illustrative in all respects and are not limiting.

Claims

1. An ester derivative compound of inotodiol represented by the following chemical formula or a pharma- ceutical acceptable salt thereof 【Chemistry 1】 [In the above formula, R １ and R ２ are each independently OH or -OC(O)-R ３ and R ３ is a linear or branched alkyl chain having 2 to 30 carbon atoms; R １ and R ２ At least one of -OC(O)-R ３ is.]

2. 2. The method for producing an ester derivative compound of inotodiol or a pharma- ceutically acceptable salt thereof according to claim 1, wherein the ester derivative compound of inotodiol is represented by the following chemical formula: 【Chemistry 2】 and a carboxyl group of a C 2 -C 30 saturated fatty acid by ester condensation reaction.

3. 3. The method of claim 2, wherein the saturated fatty acid is selected from the group consisting of caprylic acid, capric acid, lauric acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, cerotic acid, and myristic acid.

4. The method of claim 2, wherein the inotodiol and fatty acid are mixed in a molar ratio of 1:10-30.

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