Novel compound, use thereof, and method for producing novel compound

Abstract

To provide a novel compound exhibiting DPP-4 inhibitory activity and a method for producing a composition comprising the compound.SOLUTION: Provided is a compound represented by the following formula (1), obtained by a method including a step of extracting from fruits of Kochia scoparia, where X represents any one of a monosaccharide, a disaccharide, and a trisaccharide.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a novel compound, its use, and a method for producing the novel compound. [Background technology]

[0002] Mountain caviar is the fruit of the Kochia scoparia, which has recently been widely cultivated for ornamental purposes under the name Kochia. Odate City in Akita Prefecture also produces a specialty called "Tonburi" (a processed product made from the mature fruit of the Kochia), which has a crunchy texture and looks similar to caviar, earning it the nickname "caviar from the fields." Meanwhile, in China, it is called "jifushi" and is described in the Shennong Materia Medica, a book from the Later Han Dynasty, as having the following effects: "Relieves heat from the bladder, improves urination, replenishes kidney energy, and if taken over a long period of time, improves ear and eye clarity, makes the body lighter, and slows aging," and it has been used as a medicinal herb for over 2,000 years. It is believed to have been introduced to Japan from China during the Heian period, and there are records of the cultivation of broom trees in the Edo period documents "Comprehensive Agriculture Book" and "Pop Songs." Since the Heian period, the stems have been used to make brooms (hence the name broom tree), and the fruits have been eaten on special occasions such as vegetarian cuisine. In the Heisei era, a group led by Professor Emeritus Yoshikawa and Professor Matsuda of Kyoto Pharmaceutical University, a leading authority on saponin research in Japan, conducted vigorous research on the components and physiological activities of mountain caviar. Their results showed that the saponin momordin Ic is the active ingredient and suppresses blood glucose levels in a glucose tolerance test in rats (Non-Patent Document 1). Its mechanism of action is based on the inhibition of carbohydrate transport from the stomach to the small intestine (Non-Patent Documents 2 and 3) and the inhibition of glucose absorption by suppressing the sodium-dependent glucose transporter (SGLT1) in the small intestinal mucosa (Non-Patent Document 4). Therefore, unlike blood sugar control foods that inhibit α-glucosidase (e.g., Salacia, mulberry leaf extract), it is characterized by its ability to suppress the absorption of glucose, the main component of "liquid sugar" used in soft drinks and other beverages. In other words, it exhibits broad carbohydrate absorption inhibition by suppressing the absorption of oligosaccharides and glucose with lower molecular weights than maltose. The saponins in mountain caviar are oleanane-type triterpene saponins. In addition, it contains trace amounts of saponin components such as kochianosides (Non-Patent Document 5) and scoparianosides (Non-Patent Document 1). Its blood sugar level-inhibiting effect has been confirmed in rats (Non-Patent Document 5). The functional components are momordin Ic and 2'-O-glucopyranosyl momordin Ic.

[0003] In diabetes, the body's ability to produce insulin is significantly reduced, or even if insulin is produced, the insulin does not act effectively due to reduced insulin sensitivity, etc., resulting in an increase in blood glucose levels and various hyperglycemic symptoms. Currently, a widely used treatment, in addition to insulin administration, is to control blood glucose levels by administering hypoglycemic agents.

[0004] On the other hand, with the increasing number of type II diabetes patients, various treatments other than conventional insulin administration are being investigated, and among them, incretins secreted by the body have been attracting attention.

[0005] Incretins are a general term for gastrointestinal hormones known to lower blood glucose levels by stimulating insulin secretion. Among incretins, GLP-1 analogs and the like have been developed as pharmaceuticals. Furthermore, since GLP-1 is degraded and inactivated by the peptide-degrading enzyme dipeptidyl peptidase 4 (DPP4), substances that inhibit the action of DPP4 are also being developed as new diabetes treatments (Patent Document 1).

[0006] [Non-Patent Document 1] Yoshikawa M., Shimada H., Morikawa T., Yoshizumi S., Matsumura N., Murakami T., Matsuda H., Hori K., Yamahara J. Medicinal foodstuffs. VII. On the saponin constituents with glucose and alcohol absorption-inhibitory activity from a food garnish "Tonburi", the fruit of Japanese Kochia scoparia (L.) Schrad.: Structures of scoparianosides A, B, and C. Chem. Pharm. Bull., 45, 1300-1305 (1997). [Non-Patent Document 2] Matsuda H., Li Y., Yamahara J., Yoshikawa M. Inhibition of gastric emptying by triterpene saponin, momordin Ic, in mice: roles of blood glucose, capsaicin-sensitive sensory nerves, and central nervous system. J. Pharmacol. Exp. Ther., 289, 729-734 (1999). [Non-Patent Document 3] Matsuda H., Li Y., Murakami T., Yamahara J., Yoshikawa M. Structure-related inhibitory activity of oleanolic acid glycosides on gastric emptying in mice. Bioorg. Med. Chem., 7, 323-327 (1999). [Non-Patent Document 4] Matsuda H., Li Y., Murakami T., Matsumura N., Yamahara J., Yoshikawa M. Antidiabetic principles of natural medicines. III. Structure-related inhibitory activity and action mode of oleanolic acid glycosides on hypoglycemic activity. Chem. Pharm. Bull., 46, 1399-1403 (1998). [Non-patent document 5] Yoshikawa M., Dai Y., Shimada H., Morikawa T., Matsumura N., Yoshizumi S., Matsuda H., Matsuda H., Kubo M. Studies on Kochiae Fructus. II. On the saponin constituents from the fruit of Chinese Kochia scoparia (Chenopodiaceae): chemical structures of kochianosides I, II, III, and IV. Chem. Pharm. Bull., 45, 1052-1055 (1997). [Patent Document 1] Japanese Patent Application Laid-Open No. 2014-019657 DISCLOSURE OF THE INVENTION [Problem to be solved by the invention]

[0007] The above saponins are contained in the butanol-soluble fraction of the extract. On the other hand, the water-soluble fraction is present in an amount equal to or greater than that of the butanol fraction, but little research has been done on its components. Against this background, the present inventor discovered that the water-soluble fraction of mountain caviar contains three new components, isolated these three components, and examined the inhibitory activity of these new components against DPP-4, an enzyme related to diabetes. As a result, it was found that all three new components have inhibitory activity, thereby completing the present invention. That is, an object of the present invention is to obtain a novel compound from the fruit of Kochia scoparia and to provide uses thereof. [Means for solving the problem]

[0008] The features of the present invention for solving the above problems are as follows. 1. A compound represented by the following chemical formula (1): [ka] X in the above chemical formula (1) is substituted with any one of monosaccharides, disaccharides, and trisaccharides. 2. Compounds represented by the following chemical formulas (2) to (4): [ka] [ka] [ka] 3. A DPP-4 inhibitor containing the compound shown in 1 or 2 above as an active ingredient. 4. A method for producing a composition containing the compounds represented by chemical formulas (2) to (4) described in 2 above, characterized by comprising step (1) of extracting the fruits of Kochia scoparia with a polar solvent to obtain a Kochia scoparia fruit extract, and step (2) of concentrating the Kochia scoparia fruit extract after step (1). [Effects of the Invention]

[0009] The compounds of the present invention represented by chemical formulas (1) to (4) are novel and have high inhibitory activity against DPP-4. Therefore, they are useful as therapeutic agents for diabetes containing them as active ingredients, and can be used in pharmaceuticals, food compositions, etc. [Brief explanation of the drawings]

[0010] [Figure 1] FIG. 1 shows a method for isolating a novel component from Kochia spp. fruits. [Figure 2] FIG. 1 shows a chart of an HPLC chromatogram of an isolated component of a novel component. [Figure 3] FIG. 1 is a diagram showing two-dimensional NMR correlation of the compound represented by chemical formula (2). [Figure 4] FIG. 1 is a diagram showing two-dimensional NMR correlation of the compound represented by chemical formula (3). [Figure 5] FIG. 1 is a diagram showing two-dimensional NMR correlation of the compound represented by chemical formula (4). DETAILED DESCRIPTION OF THE INVENTION

[0011] The present invention will be described in detail below. The present invention is a compound represented by the following chemical formula (1). [ka] In the above chemical formula, X is substituted with either a monosaccharide, disaccharide, or trisaccharide. Examples of monosaccharides include ribose, arabinose, xylose, lyxose, psicose, fructose, sorbose, tagatose, allose, altrose, glucose, mannose, gulose, idose, galactose, talose, apiose, and rhamnose, with glucose and galactose being preferred. Further, examples of disaccharides include sucrose, trehalose, and lactose, and those composed of any two of galactose, apiose, and rhamnose are preferred, but are not limited to these. Examples of trisaccharides include raffinose, melezitose, and maltotriose, and those composed of galactose, apiose, and rhamnose are preferred, but are not limited to these.

[0012] The present invention relates to compounds represented by the following chemical formulas (2) to (4). [ka] [ka] [ka]

[0013] The compounds represented by the above chemical formulas (1) to (4) can be isolated by extracting the fruits of Kochia scoparia with a solvent.

[0014] The extraction solvent may be a polar solvent such as water, methanol, ethanol, isopropyl alcohol, 1,3-butylene glycol, ethylene glycol, propylene glycol, glycerin, ethyl acetate, etc. Two or more of these solvents may be mixed. Preferably, water, ethanol or a mixture thereof, such as aqueous ethanol, is used as the extraction solvent, which allows for efficient extraction of the active ingredients.

[0015] When water is used as the extraction solvent, the type of water is not particularly limited, and tap water, distilled water, mineral water, alkaline ionized water, deep sea water, etc. can be used.

[0016] When aqueous ethanol is used as the extraction solvent, the ethanol concentration is not particularly limited, but it is particularly preferable that the ethanol concentration be 10 to 90% (wt / wt), and preferably 20 to 80% (wt / wt).

[0017] The extraction temperature should be 20 to 80° C., preferably about 40 to 50° C. If the extraction temperature is too low, it becomes difficult to extract the active ingredients, and if the extraction temperature is too high, the active ingredients will decompose, resulting in a decrease in physiological activity (health functionality).

[0018] As the extraction method, any method such as stirring extraction, continuous extraction, immersion extraction, countercurrent extraction, and supercritical extraction can be employed, and any apparatus can be used at room temperature or under reflux heating.

[0019] Specifically, the extraction method involves placing the crushed raw material (fruit of Kochia scoparia) in a treatment tank filled with an extraction solvent and stirring to extract the active ingredients. For example, when using aqueous ethanol as the extraction solvent, an amount of extraction solvent approximately 2 to 100 times (by weight) the amount of extraction raw material is used, and extraction is carried out for approximately 30 minutes to 2 hours. After the active ingredients have been extracted into the solvent, the extract is obtained by filtering to remove the extraction residue.

[0020] Thereafter, the extract is subjected to treatments such as dilution, concentration, purification, and drying according to conventional methods to obtain the agent according to the present invention. The purification method may be such that the extract is passed through a synthetic adsorption resin, a gel filtration resin, or the like to adsorb the active ingredient, and then the active ingredient is eluted with methanol, ethanol, or the like to concentrate it.

[0021] After obtaining an extract from the fruits of Kochia scoparia using the above-mentioned method, the extract can be obtained by solid-liquid separation from the residue using conventional methods such as filtration and centrifugation. In the present invention, the obtained extract can be used directly as a DPP-4 inhibitor, but since the activity may be low, it can also be used in the form of an extract or powder by appropriate concentration or solvent removal. Furthermore, active fractions can be isolated from the obtained extract by solvent fractionation using one or more organic solvents such as methanol, ethanol, propanol, butanol, dichloromethane, chloroform, ethyl acetate, toluene, hexane, benzene, and acetone. Furthermore, if necessary, purification can be performed using one or a combination of suitable separation and purification methods such as alumina column chromatography, silica gel chromatography, gel filtration chromatography, ion exchange chromatography, hydrophobic chromatography, and high-performance liquid chromatography.

[0022] The compounds represented by the chemical formulas (1) to (4) can be isolated and identified from the fruits of Kochia scoparia, specifically according to the description in the Examples.

[0023] As described above, the compounds represented by chemical formulas (1) to (4) of the present invention can be isolated and purified from natural products, but the method for producing the compounds is not limited thereto. The compounds may be synthesized based on known chemical synthesis methods, or they may be produced by subjecting substances obtained from natural products as raw materials to treatment such as reaction.

[0024] As shown in the test examples described below, the compounds of the present invention represented by chemical formulas (1) to (4) have DPP-4 inhibitory activity and are useful as active ingredients of DPP-4 inhibitors. Specifically, the DPP-4 inhibitors of the present invention are useful as therapeutic agents for diabetes (including those at risk of diabetes).

[0025] The compounds represented by chemical formulas (1) to (4) of the present invention can be used as ingredients in various foods and beverages. Examples of foods and beverages include general foods such as confectioneries (gum, candy, caramel, chocolate, cookies, snacks, jellies, gummies, tablets, etc.), noodles (soba, udon, ramen, etc.), dairy products (milk, ice cream, yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, beverages (juice, coffee, black tea, green tea, carbonated drinks, sports drinks, etc.), as well as health foods (tablets, capsules, etc.), and nutritional supplements (nutritional drinks, etc.). The agent of the present invention can be appropriately incorporated into these foods and beverages. In this case, isolated compounds of the above chemical formulas (1) to (4) may be added, or an extract containing the above chemical formulas (1) to (4) as active ingredients, such as an extract of the fruit of Kochia scoparia, may be used.

[0026] These foods and beverages can contain various ingredients depending on their type, such as glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid esters, polyglycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, gum arabic, carrageenan, casein, gelatin, pectin, agar, B vitamins, nicotinamide, calcium pantothenate, amino acids, calcium salts, colorants, flavorings, and preservatives.

[0027] Specifically, the agent of the present invention can be spray-dried or freeze-dried together with powdered cellulose, and then formed into a powder, granules, tablets, or solution, which can be easily incorporated into foods and beverages (e.g., instant foods). The agent of the present invention can also be dissolved in, for example, fats and oils, ethanol, glycerin, or a mixture thereof to form a liquid, which can then be added to beverages or solid foods. If necessary, the agent can also be mixed with a binder such as gum arabic or dextrin to form a powder or granules, which can then be added to beverages or solid foods.

[0028] When the agent of the present invention is applied to food and drink, the amount of active ingredient added is preferably 1 to 20 wt % in total relative to the food and drink, since the main purpose is disease prevention and health maintenance.

[0029] The compounds of the present invention represented by chemical formulas (1) to (4) may be used as raw materials for pharmaceuticals (including pharmaceuticals and quasi-drugs). Pharmaceuticals can be produced by appropriately blending the agent of the present invention with raw materials for pharmaceutical preparations. Examples of pharmaceutical raw materials that can be incorporated into the agent of the present invention include excipients (glucose, lactose, sucrose, sodium chloride, starch, calcium carbonate, kaolin, crystalline cellulose, cocoa butter, hardened vegetable oil, kaolin, talc, etc.), binders (distilled water, saline, ethanol water, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.), disintegrants (sodium alginate, agar, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, stearate monoglyceride, starch, lactose, gum arabic powder, gelatin, ethanol, etc.), disintegration inhibitors (sucrose, stearin, cocoa butter, hydrogenated oil, etc.), absorption enhancers (quaternary ammonium base, sodium lauryl sulfate, etc.), adsorbents (glycerin, starch, lactose, kaolin, bentonite, silicic acid, etc.), lubricants (purified talc, stearates, polyethylene glycol, etc.). In this case, the compounds of the above chemical formulas (1) to (4) may be isolated and added, or an extract containing the compounds of the above chemical formulas (1) to (4) as active ingredients, such as an extract of the fruit of Kochia scoparia, may be used.

[0030] The compounds of the present invention represented by chemical formulas (1) to (4) can generally be administered orally in the form of tablets, pills, soft or hard capsules, fine granules, powders, granules, liquids, etc., but may also be administered parenterally. When administered parenterally, they can be administered in the form of a solution or in the form of a dispersant, suspending agent, stabilizer, etc., by local tissue administration, intradermal, subcutaneous, intramuscular, or intravenous injection. They may also be in the form of suppositories, etc.

[0031] The dosage may vary depending on the administration method, the condition, the age of the patient, etc., but typically, adults can be given 1 to 500 mg of the active ingredient per day, and children can be given 0.5 to 300 mg. The compounding ratio of the compounds represented by chemical formulas (1) to (4) can be varied depending on the dosage form, but is typically about 0.3 to 15.0 wt% when administered orally or via mucosal absorption, and about 0.01 to 1 wt% when administered parenterally. The dosage varies depending on various conditions, and a smaller amount than the above may be sufficient, or an amount exceeding the range may be required. In this case, the compounds represented by chemical formulas (1) to (4) may be added in their isolated form, or an extract containing the compounds represented by chemical formulas (1) to (4) as active ingredients, such as an extract of the fruit of Kochia scoparia, may be used. [Example]

[0032] Examples of the present invention will be described below. Note that the examples shown below are provided to confirm the various actions and effects of the agent of the present invention obtained by the present invention, and the scope of the present invention is not limited to these products and manufacturing methods.

[0033] Example: Isolation of a novel component The novel component was isolated as shown in Figure 1. A 65% ethanol extract (1674 g, including water) of the fruit of Chinese kochia (Kochia scoparia) was suspended in water (9 L) and washed successively with ethyl acetate (10 L, twice) and butanol (10 L, twice). The remaining liquid was concentrated to obtain an aqueous fraction (196 g). The entire aqueous fraction was applied to an ODS (Fuji Silysia, 730 g) column, washed with 10% methanol (2 L) and 20% methanol (2 L), and then eluted with 30% methanol (2 L) and 40% methanol (2 L). The resulting solution was concentrated to give a 30% methanol fraction (4.5 g) and a 40% methanol fraction (6.55 g). Next, the 40% methanol fraction (2.75 g) was fractionated by HPLC [column: Cosmosil C18 PAQ (inner diameter 20 mm × length 250 mm, Nacalai Tesque), mobile phase: 40% methanol] and separated into fractions (Fr.) 1 to 5. The resulting Fraction 4 was purified by HPLC [Column: Inertsil NH2 (inner diameter 20 mm × length 250 mm (GL Sciences)), Mobile phase: 80% acetonitrile] and fractionated into Fractions 4-1 to 4-3. Fraction 4-2 was purified by HPLC [Column: Capcellpak C18 SG120 (inner diameter 20 mm × length 250 mm, Osaka Soda), Mobile phase: 40% methanol] to obtain kochiaflavonoside A (15.1 mg). Fraction 5 was purified by HPLC [column: Inertsil NH2 (inner diameter 20 mm × length 250 mm), mobile phase: 80% acetonitrile] to yield fractions 5-1 to 5-3. Fraction 5-2 was purified by HPLC [column: Capcellpak C18 SG120 (inner diameter 20 mm × length 250 mm), mobile phase: 40% methanol] to yield kochiaflavonoside B (13 mg). Fraction 5-1 was purified by HPLC [column: Capcellpak C18 SG120 (inner diameter 20 mm × length 250 mm), mobile phase: 40% methanol] to yield kochiaflavonoside C (3 mg). Figure 2 shows the HPLC charts of the obtained single substances, which show that they are all single substances.

[0034] Identification of new components The obtained components were analyzed by optical rotation, mass spectrometry, infrared absorption, UV absorption spectrometry, 1 H and 13 C-NMR spectra were measured. 1. The compound represented by chemical formula (2) (kochiaflavonoside A) 1) Physicochemical data Yellow powder. [α] D 25 -57.4° (c=1.02, MeOH). UV [MeOH, nm, (log ε)]:205 (3.75), 255 (3.51), 271 (3.40), 296 (3.32), 335 (3.52), 361 (3.38). High resolution ESI-MS Calcd for C 48 H 56 O 29 Na (M+Na) + : ; Found: 1119.2777, IR (KBr, cm -1 ): 3370, 2920, 2850, 1701, 1597, 1271, 1193, 1074. 1H-NMR (DMSO-d6, 800 MHz) δ: 3.19 (1H×2, m, 4''''', 5'''''-H), 3.23 (1H, m, 4''''-H), 3.28 (1H, dd, J=6.0, 11.0 Hz, 6''a-H), 3.33 (1H, m, 5''-H), 3.39 (1H, brd, 5a'''-H), 3.40 (1H, brd, 6b''-H), 3.41 (1H, m, 3'''''-H), 3.42 (1H, m, 3''''-H), 3.43 (1H, brd, 6a'''''-H), 3.44 (1H×2, m, 5b''', 5''''-H), 3.49 (1H, d, J=9.6 Hz, 4a'''-H), 3.50 (1H, dd, J=5.0, 11.0 Hz, 6a''''-H), 3.57 (1H×2, m, 3'', 6b''''-H), 3.59 (1H, dd, J=7.7, 8.8 Hz, 2''''-H), 3.64 (1H, m, 4''-H), 3.66 (1H, brd, 6b'''''-H), 3.78 (1H, dd, J=7.7, 9.3 Hz, 2''-H), 3.81 (1H, d, J=1.4 Hz, 2'''-H), 3.81 (3H, s, -OCH3), 3.84 (1H, d, J=9.6 Hz, 4b'''-H), 4.61 (1H, dd, J=8.0, 9.3 Hz, 2'''''-H), 4.90 (1H, d, J=8.0 Hz, 1'''''-H), 5.20 (1H, d, J=7.7 Hz, 1''''-H), 5.32 (1H, d, J=1.4 Hz, 1'''-H), 5.58 (1H, d, J=7.7 Hz, 1''-H), 6.40 (1H, d, J=15.9 Hz, 8''''''-H), 6.42 (1H, d, J=2.2 Hz, 6-H), 6.73 (1H, d, J=2.2 Hz, 8-H), 6.78 (1H, d, J=8.2 Hz, 5''''''-H), 6.81 (1H, d, J=8.5 Hz, 5'-H), 7.08 (1H, dd, J=1.9, 8.2 Hz, 6''''''-H), 7.25 (1H, d, J=1.9 Hz, 2''''''-H), 7.51 (1H, d, J=15.9 Hz, 7''''''-H), 7.53 (1H, d, J=2.2 Hz, 2'-H), 7.76 (1H, dd, J=2.2, 8.5 Hz, 6'-H), 12.68 (brs, 4-OH). 13 C-NMR (DMSO-d6, 200 MHz) δ c : 155.9 (C-2), 133.6 (C-3), 177.6 (C-4), 160.9 (C-5), 99.5 (C-6), 162.6 (C-7), 94.3 (C-8), 156.4 (C-9), 105.6 (C-10), 121.2 (C-1'), 116.0 (C-2'), 145.0 (C-3'), 148.7 (C-4'), 115.2 (C-5'), 122.5 (C-6'), 99.0 (C-1''), 75.0 (C-2''), 73.9 (C-3''), 68.4 (C-4''), 75.8 (C-5''), 60.1 (C-6''), 108.9 (C-1'''), 76.3 (C-2'''), 79.2 (C-3'''), 74.0 (C-4'''), 64.5 (C-5'''), 97.8 (C-1''''), 79.6 (C-2''''), 76.2 (C-3''''), 70.2 (C-4''''), 76.8 (C-5''''), 60.8 (C-6''''), 100.4 (C-1'''''), 74.1 (C-2'''''), 74.6 (C-3'''''), 69.9 (C-4'''''), 76.9 (C-5'''''), 60.6 (C-6'''''), 125.9 (C-1''''''), 111.5 (C-2''''''), 148.0 (C-3''''''), 149.2 (C-4''''''), 115.7 (C-5''''''), 122.8 (C-6''''''), 144.5 (C-7''''''), 115.3 (C-8''''''), 165.9 (C-9''''''), 55.9 (-OCH3). Positive-ion ESI-MS: m / z 1119 (M+Na) + .

[0035] 2) Two-dimensional NMR HMQC ( 1 H-detected multiple quantum coherence spectrum), HMBC ( 1 H-detected multi-bond heteronuclear multiple quantum coherence spectrum) and 1 H- 1 As a result of H COSY (Correlation spectroscopy) analysis, the correlation between protons and carbons shown by the arrows in Figure 3, the correlation between protons and protons shown by the thick lines, and the spatial proximity between protons and protons shown by the wavy arrows were observed.

[0036] 3) Identification of acyl groups and sugars First, to identify the acyl group, alkaline hydrolysis was performed as follows. Kochiaflavonoside A (5.0 mg) was dissolved in 0.5% potassium hydroxide solution (500 μL) and reacted at 40°C for 1 hour. After the reaction, the solution was diluted with Dowex HCR W2 (H + The solution was neutralized with ethanol and filtered through a cotton plug. The solution was subjected to reversed-phase HPLC [column: Cosmosil πNAP, 250 mm × 4.6 mm inner diameter (Nacalai Tesque)], eluent: acetonitrile: 1% acetic acid = 25:75, detection: UV (254 nm), flow rate: 1 mL / min]. The retention time (t R The acyl group was identified as ferulic acid by comparing with the acetylcholinesterase activity (9.5 min). Next, to identify the sugars, acid hydrolysis of the desacyl forms after acyl decomposition was carried out as follows. The reaction product after acyl decomposition was dissolved in 1.0 M hydrochloric acid (1.0 mL) and reacted at 80°C for 3 hours. After cooling, the reaction mixture was filtered with Amberlite IRA-400 (OH). -The sugar fraction was subjected to HPLC [column: Kaseisorb LC NH2-60-5, 250 mm × 4.6 mm inner diameter (Tokyo Kasei), eluent: acetonitrile:water = 85:15, detection: optical rotation detection, flow rate: 1 mL / min], and the retention time [t R : D-apiose; 8.9 min (positive), D-glucose; 13.8 min (positive), D-galactose; 14.8 min (positive)]. Based on the above physicochemical data, the structure of this novel component was determined to be the compound (kochiaflavonoside A) shown in chemical formula (2).

[0037] 2. Compound represented by chemical formula (3) (kochiaflavonoside B) 1) Physicochemical data Yellow powder. [α] D 25 -50.1° (c=0.1, MeOH). UV [MeOH, nm, (log ε)]:206 (3.62), 256 (3.39), 272 (3.27), 298 (3.19), 333 (3.38), 374 (3.15). High resolution ESI-MS Calcd for C 49 H 58 O 29 Na (M+Na) + ; Found: 1133.2958, IR (KBr, cm -1 ): 3383, 2930, 1699, 1653, 1597, 1506, 1273, 1072. 1H-NMR (DMSO-d6, 800 MHz) δ: 1.07 (3H, d, J=6.1 Hz, 6'''-H), 3.11 (1H, dd, J=9.2, 9.5 Hz, 4'''-H), 3.20 (1H, dd, J=9.2, 9.5 Hz, 4'''''-H), 3.20 (1H, m, 5'''''-H), 3.24 (1H, dd, J=8.9, 9.5 Hz, 4''''-H), 3.27 (1H, dd, J=3.9, 10.1 Hz, 6''a-H), 3.32 (1H, dd, J=3.4, 9.5 Hz, 3'''-H), 3.38 (1H, qd, J=6.1, 9.2 Hz, 5'''-H), 3.40 (1H, dd, J=9.2, 9.5 Hz, 4''''-H), 3.40 (1H, dd, J=9.2, 9.5 Hz, 4'''''-H), 3.40 (1H, m, 4''-H), 3.42 (1H, dd, J=1.2, 3.4 Hz, 2'''-H), 3.43 (1H×2, m, 5'''', 6'''''a-H), 3.51 (1H, dd, J=4.9, 11.6 Hz, 6''''a-H), 3.56 (1H, ddd, J=3.9, 6.4, 9.7 Hz, 5''-H), 3.57 (1H, m, 2''-H), 3.58 (1H, dd, J=7.3, 9.2 Hz, 2''''-H), 3.58 (1H, m, 6''''b-H), 3.61 (1H, dd, J=2.4, 9.7 Hz, 4''-H), 3.62 (1H, dd, J=6.4, 10.1 Hz, 6''b-H), 3.66 (1H, brd, 6'''''b-H), 3.80 (3H, s, -OCH3), 4.43 (1H, d, J=1.2 Hz, 1'''-H), 4.61 (1H, dd, J=7.9, 9.2 Hz, 2'''''-H), 4.91 (1H, d, J=7.9 Hz, 1'''''-H), 5.21 (1H, d, J=7.3 Hz, 1''''-H), 5.33 (1H, d, J=7.6 Hz, 1''-H), 6.37 (1H, d, J=15.9 Hz, 8''''''-H), 6.43 (1H, d, J=2.1 Hz, 6-H), 6.73 (1H, d, J=2.1 Hz, 8-H), 6.77 (1H, d, J=8.2 Hz, 5''''''-H), 6.83 (1H, d, J=8.2 Hz, 5'-H), 7.05 (1H, dd, J=1.8, 8.2 Hz, 6''''''-H), 7.22 (1H, d, J=1.8 Hz, 2''''''-H), 7.49 (1H, d, J=15.9 Hz, 7''''''-H), 7.56 (1H, d, J=2.1 Hz, 2'-H), 7.64 (1H, dd, J=2.1, 8.2 Hz, 6'-H), 12.53 (brs, 4-OH). 13 C-NMR (DMSO-d6, 200 MHz) δ c: 157.1 (C-2), 133.9 (C-3), 177.7 (C-4), 160.9 (C-5), 99.7 (C-6), 162.7 (C-7), 94.5 (C-8), 156.1 (C-9), 105.7 (C-10), 121.1 (C-1'), 116.4 (C-2'), 144.9 (C-3'), 148.8 (C-4'), 115.3 (C-5'), 122.1 (C-6'), 102.2 (C-1''), 71.3 (C-2''), 73.2 (C-3''), 68.2 (C-4''), 73.7 (C-5''), 65.3 (C-6''), 100.1 (C-1'''), 70.5 (C-2'''), 70.8 (C-3'''), 72.1 (C-4'''), 68.4 (C-5'''), 18.0 (C-6'''), 97.8 (C-1''''), 79.6 (C-2''''), 76.0 (C-3''''), 70.2 (C-4''''), 76.8 (C-5''''), 60.8 (C-6''''), 100.2 (C-1'''''), 74.1 (C-2'''''), 74.6 (C-3'''''), 69.9 (C-4'''''), 76.9 (C-5'''''), 60.6 (C-6'''''), 125.9 (C-1''''''), 111.4 (C-2''''''), 148.0 (C-3''''''), 149.3 (C-4''''''), 115.7 (C-5''''''), 122.8 (C-6''''''), 144.5 (C-7''''''), 115.3 (C-8''''''), 165.9 (C-9''''''), 55.9 (-OCH3). Positive-ion ESI-MS: m / z 1133 (M+Na) + .

[0038] 2) Two-dimensional NMR HMQC ( 1 H-detected multiple quantum coherence spectrum), HMBC ( 1H-detected multi-bond heteronuclear multiple quantum coherence spectrum) and 1 H- 1 As a result of H COSY (Correlation spectroscopy) analysis, the correlation between protons and carbons shown by the arrows in Figure 4, the correlation between protons and protons shown by the thick lines, and the spatial proximity between protons and protons shown by the wavy arrows were observed.

[0039] 3) Identification of acyl groups and sugars First, to identify the acyl group, alkaline hydrolysis was performed as follows. Kochiaflavonoside B (2.7 mg) was dissolved in 0.5% potassium hydroxide solution (500 μL) and reacted at 40°C for 1 hour. After the reaction, the solution was diluted with Dowex HCR W2 (H + The solution was neutralized with ethanol and filtered through a cotton plug. The solution was subjected to reversed-phase HPLC [column: Cosmosil πNAP, 250 mm × 4.6 mm inner diameter (Nacalai Tesque)], eluent: acetonitrile: 1% acetic acid = 25:75, detection: UV (254 nm), flow rate: 1 mL / min]. The retention time (t R The acyl group was identified as ferulic acid by comparing with the acetylcholinesterase activity (9.5 min). Next, to identify the sugars, acid hydrolysis of the desacyl forms after acyl decomposition was carried out as follows. The reaction product after acyl decomposition was dissolved in 1.0 M hydrochloric acid (1.0 mL) and reacted at 80°C for 3 hours. After cooling, the reaction mixture was filtered with Amberlite IRA-400 (OH). - The sugar fraction was subjected to HPLC [column: Kaseisorb LC NH2-60-5, 250 mm × 4.6 mm inner diameter (Tokyo Kasei), eluent: acetonitrile:water = 85:15, detection: optical rotation detection, flow rate: 1 mL / min], and the retention time [t RThe kinetics of the enzymes were identified by comparing the enzyme activity with the enzymes listed below: L-rhamnose; 7.4 min (negative), D-glucose; 13.8 min (positive), D-galactose; 14.8 min (positive). Based on the above physicochemical data, the structure of this novel component was determined to be the compound (kochiaflavonoside B) shown in chemical formula (3).

[0040] 3. Compound represented by chemical formula (4) (kochiaflavonoside C) 1) Physicochemical data Yellow powder. [α] D 25 -81.2° (c=0.05, MeOH). UV [MeOH, nm, (log ε)]:206 (3.37), 256 (3.15), 272 (3.04), 298 (2.94), 333 (3.12), 372 (2.92). High resolution ESI-MS Calcd for C 43 H 48 O 25 Na (M+Na) + : ; Found: 987.2377, IR (KBr, cm -1 ): 3383, 2920, 1699, 1653, 1597, 1506, 1273, 1072. 1H-NMR (DMSO-d6, 800 MHz) δ: 3.20 (1H×2, m, 3'''', 4''''-H), 3.24 (1H, dd, J=8.9, 9.5 Hz, 4'''-H), 3.31 (1H, dd, J=6.1, 10.7 Hz, 6''a-H), 3.34 (1H, brdd, 5''-H), 3.38 (1H, dd, J=3.4, 9.5 Hz, 3''-H), 3.41 (1H, dd, J=8.8, 9.5 Hz, 3'''-H), 3.41 (1H, dd, J=9.2, 9.5 Hz, 3''''-H), 3.43 (1H, ddd, J=1.8, 8.8, 9.5 Hz, 5'''-H), 3.43 (1H, m, 6''''a-H), 3.46 (1H, dd, J=5.8, 10.7 Hz, 6''b-H), 3.51 (1H, dd, J=4.9, 11.6 Hz, 6'''a-H), 3.58 (1H, dd, J=7.6, 9.5 Hz, 2''-H), 3.58 (1H, dd, J=1.8, 11.6 Hz, 6'''-H), 3.59 (1H, dd, J=7.6, 8.8 Hz, 2'''-H), 3.65 (1H, m, 4''-H), 3.65 (1H, brd, 6''''b-H), 3.80 (3H, s, -OCH3), 4.61 (1H, dd, J=8.2, 9.5 Hz, 2''''-H), 4.91 (1H, d, J=8.2 Hz, 1''''-H), 5.21 (1H, d, J=7.6 Hz, 1'''-H), 5.37 (1H, d, J=7.6 Hz, 1''-H), 6.38 (1H, d, J=15.9 Hz, 8'''''-H), 6.43 (1H, d, J=2.1 Hz, 6-H), 6.74 (1H, d, J=2.1 Hz, 8-H), 6.77 (1H, d, J=8.2 Hz, 5'''''-H), 6.82 (1H, d, J=8.2 Hz, 5'-H), 7.05 (1H, dd, J=1.8, 8.2 Hz, 6'''''-H), 7.22 (1H, d, J=2.1 Hz, 2'''''-H), 7.49 (1H, d, J=15.9 Hz, 7'''''-H), 7.56 (1H, d, J = 2.1 Hz, 2'-H), 7.65 (1H, dd, J = 2.1, 8.2 Hz, 6'-H), 12.58 (brs, 4-OH). 13 C-NMR (DMSO-d6, 200 MHz) δ c : 157.1 (C-2), 133.9 (C-3), 177.7 (C-4), 160.9 (C-5), 99.6 (C-6), 162.6 (C-7), 94.4 (C-8), 156.0 (C-9), 105.7 (C-10), 121.0 (C-1'), 116.3 (C-2'), 144.9 (C-3'), 148.9 (C-4'), 115.3 (C-5'), 122.1 (C-6'), 101.9 (C-1''), 71.4 (C-2''), 73.3 (C-3''), 68.1 (C-4''), 75.9 (C-5''), 60.2 (C-6''), 97.8 (C-1'''), 79.6 (C-2'''), 76.0 (C-3'''), 70.2 (C-4'''), 76.8 (C-5'''), 60.8 (C-6'''), 100.2 (C-1''''), 74.1 (C-2''''), 74.6 (C-3''''), 69.9 (C-4''''), 76.9 (C-5''''), 60.6 (C-6''''), 125.9 (C-1'''''), 111.4 (C-2'''''), 148.0 (C-3'''''), 149.3 (C-4'''''), 115.7 (C-5'''''), 122.1 (C-6'''''), 144.5 (C-7'''''), 115.3 (C-8'''''), 165.9 (C-9'''''), 55.8 (-OCH3). Positive-ion ESI-MS: m / z 987 (M+Na) + .

[0041] 2) Two-dimensional NMR HMQC ( 1 H-detected multiple quantum coherence spectrum), HMBC ( 1H-detected multi-bond heteronuclear multiple quantum coherence spectrum) and 1 H- 1 As a result of H COSY (Correlation Spectroscopy) analysis, the correlation between protons and carbons shown by the arrows in Figure 5, the correlation between protons and protons shown by the thick lines, and the spatial proximity between protons and protons shown by the wavy arrows were observed.

[0042] 3) Identification of acyl groups and sugars First, to identify the acyl group, alkaline hydrolysis was performed as follows. Kochiaflavonoside C (2.3 mg) was dissolved in 0.5% potassium hydroxide solution (500 μL) and reacted at 40°C for 1 hour. After the reaction, the solution was diluted with Dowex HCR W2 (H + The solution was neutralized with ethanol and filtered through a cotton plug. The solution was subjected to reversed-phase HPLC [column: Cosmosil πNAP, 250 mm × 4.6 mm inner diameter (Nacalai Tesque)], eluent: acetonitrile: 1% acetic acid = 25:75, detection: UV (254 nm), flow rate: 1 mL / min]. The retention time (t R The acyl group was identified as ferulic acid by comparing with the acetylcholinesterase activity (9.5 min). Next, to identify the sugars, acid hydrolysis of the desacyl forms after acyl decomposition was carried out as follows. The reaction product after acyl decomposition was dissolved in 1.0 M hydrochloric acid (1.0 mL) and reacted at 80°C for 3 hours. After cooling, the reaction mixture was filtered with Amberlite IRA-400 (OH). - The sugar fraction was subjected to HPLC [column: Kaseisorb LC NH2-60-5, 250 mm × 4.6 mm inner diameter (Tokyo Kasei), eluent: acetonitrile:water = 85:15, detection: optical rotation detection, flow rate: 1 mL / min], and the retention time [t R: D-glucose; 13.8 min (positive), D-galactose; 14.8 min (positive)]. Based on the above physicochemical data, the structure of this novel component was determined to be the compound (kochiaflavonoside C) represented by chemical formula (4).

[0043] Test example: DPP-4 inhibitory activity test of new ingredients Insulin is a hormone that lowers blood sugar levels and is secreted by the pancreas. The body also contains incretins (a general term for gastrointestinal hormones such as GLP-1) that stimulate insulin secretion after food ingestion. GLP-1 is one of the incretins and is broken down in the body by an enzyme called DPP-4 (dipeptidyl peptidase-4). Therefore, inhibiting DPP-4 can prevent the weakening of GLP-1's action. DPP-4 inhibitors, a diabetes treatment, inhibit DPP-4 to enhance the activity of GLP-1 and lower blood sugar levels by promoting insulin secretion when blood sugar levels are high. Therefore, the DPP-4 inhibitory activity of the obtained novel compounds was examined. The test was performed using Cayman Chemical's DPP(IV) Inhibitor Screening Assay Kit. Sitagliptin, a known DPP-4 inhibitor, was also evaluated as a positive control. The results are shown in Table 1 below.

[0044] [Table 1]

[0045] As a result of the test, as shown in Table 1, the inhibition rate of all three novel compounds improved in a concentration-dependent manner, and the IC50, the concentration at which the inhibition rate reached 50%, was 100 (μg / mL) or less. This confirmed that the compounds represented by chemical formulas (2) to (4) have DPP-4 inhibitory activity, and therefore are useful as DPP-4 inhibitors.

[0046] Measurement results and effects of the embodiment

[0047] The DPP-4 inhibitor according to the present invention (containing the compounds represented by the chemical formulas (2) to (4)) Kochia scoparia fruit The following are examples of blends of the extracts. Note that the following blend examples do not limit the scope of the present invention. Formulation example 1: Chewing gum Sugar 53.0wt% Gum base 20.0 Glucose 10.0 Starch syrup 16.0 Fragrance 0.5 DPP-4 inhibitor 0.5 100.0wt%

[0048] Mixing example 2: Gummies Reduced starch syrup 40.0wt% Granulated sugar 20.0 Glucose 20.0 Gelatin 4.7 Wednesday 9.68 Yuzu juice 4.0 Yuzu flavor 0.6 Dye 0.02 DPP-4 inhibitor 1.0 100.0wt%

[0049] Mixing example 3: Candy Sugar 50.0wt% Starch syrup 33.0 Wednesday 14.4 organic acid 2.0 Fragrance 0.2 DPP-4 inhibitor 0.4 100.0wt%

[0050] Blending example 4: Yogurt (hard / soft) Milk 41.5wt% Skim milk powder 5.8 Sugar 8.0 Agar 0.15 Gelatin 0.1 Lactic acid bacteria 0.005 DPP-4 inhibitor 0.4 fragrance trace amount water residue 100.0wt%

[0051] Formulation example 5: Soft drink High fructose corn syrup 30.0wt% Emulsifier 0.5 DPP-4 inhibitor 0.3 Fragrance (appropriate amount) Purified water remainder 100.0wt%

[0052] Formulation example 6: Tablet confectionery Sugar 76.4wt% Glucose 19.0 Sucrose fatty acid ester 0.2 DPP-4 inhibitor 0.5 Purified water 3.9 100.0wt%

[0053] Formulation example 7: Soft capsule Brown rice germ oil 47.0wt% Yuzu seed oil 40.0 Emulsifier 12.0 DPP-4 inhibitor 1.0 100.0wt%

[0054] Formulation example 8: Tablets Lactose 54.0wt% Microcrystalline cellulose 30.0 Starch decomposition product 10.0 Glycerin fatty acid ester 5.0 DPP-4 inhibitor 1.0 100.0wt% [Industrial Applicability]

[0055] As described above, the present invention can provide a novel compound and a therapeutic agent for diabetes using the same.

Claims

1. A compound represented by the following chemical formula (1): 【Chemical 1】 X in the above chemical formula (1) is substituted with any one of monosaccharides, disaccharides, and trisaccharides.

2. Compounds represented by the following chemical formulas (2) to (4): 【Chemistry 2】 【Chemistry 3】 【Chemistry 4】

3. A DPP-4 inhibitor comprising the compound of claim 1 or 2 as an active ingredient.

4. 3. A method for producing a composition containing a compound represented by any one of chemical formulas (2) to (4) according to claim 2, characterized in that it comprises: a step (1) of extracting the fruit of Kochia scoparia with a polar solvent to obtain a Kochia scoparia fruit extract; and a step (2) of concentrating the Kochia scoparia fruit extract after the step (1).

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