JP2008174457A - 2-(2-METHYLPROPANOYL)PHLOROGLUCINOL-1,5-DI-O-beta-D-GLUCOPYRANOSIDE HAVING ANTIOXIDANT ACTION - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new compound having an antioxidant action and usable as medicines, cosmetics, foods, drinks and degradation prevention agents effective for the prevention and treatment of various diseases hindering the health maintenance such as adult diseases and malignant rheumatoid arthritis and cosmetic troubles e.g. lesion such as scald, acne, and pigmented spot. <P>SOLUTION: The compound is 2-(2-methylpropanoyl)phloroglucinol-1,5-di-O-β-D-glucopyranoside expressed by chemical formula [1] or its pharmacologically allowable salt. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention has an anti-oxidant effect and is effective in preventing and treating various diseases that are impaired in health maintenance such as adult diseases and malignant rheumatoid arthritis, trauma such as burns, acne and stains, and other cosmetic disorders. The present invention provides novel 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside having an antioxidant action as a cosmetic and food / beverage product and serving as a deterioration inhibitor.

  In recent years, it has been clarified that lipid peroxides generated from active oxygen and unsaturated lipids have adverse effects on the body such as aging, adult diseases, and intractable diseases. For so-called antioxidants, and atherosclerosis, hyperlipidemia, cerebral infarction caused by them, malignant rheumatism, Behcet's disease, Crohn's disease, ulcerative colitis, hepatitis, gastritis, Parkinson's disease, etc. Some of their applications as therapeutic agents for diseases caused by chemical substances such as agricultural chemicals, photosensitivity, and radiation damage have been attempted. In addition to being a member of active oxygen and lipid peroxide stains, freckles, acne, skin ulcers, etc., it has also been known to degrade food and cause a decline in taste quality and nutrition, but these are still The present condition is that the substance which can fully solve a problem is not found.

  Examples of antioxidant compounds include water-soluble antioxidant compounds such as vitamin C (ascorbic acid), fat-soluble antioxidant compounds such as vitamin E (α-tocophenol), and BHA (3-tert-butyl-4-hydroxyanisole). Synthetic compounds such as) are known. However, since the synthetic compound BHA has been reported to be carcinogenic, it is desirable to ingest ingredients from food materials as raw materials from the viewpoint of safety. When a useful antioxidant is invented, it has utility value as a pharmaceutical, cosmetic, health food, food additive and the like.

  The present inventors searched for a compound derived from a natural product and having an antioxidant action, and developed a novel 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-derived from hops. It has been found that D-glucopyranoside has an excellent antioxidant action, and the present invention has been completed.

Although various fluoroacylphenone glycosides are disclosed in Patent Document 1 and Non-Patent Documents 1 to 3, 2- (2-methylpropanoyl) phloroglucinol-1,5-di- of the present invention is disclosed. O-β-D-glucopyranoside has a different structure.
WO 2006/093202 pamphlet J. Nat. Prod. 2005, 68, 1545-1548 Chinese Chemical Letters Vol. 15, No. 3, pp313-315, 2004 Planta Med 2004: 70: 1128-1134

The present invention relates to the following.
(1) Chemical formula [1]

2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside or a pharmaceutically acceptable salt thereof, wherein
(2) The 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside according to (1), which is obtained from a plant belonging to the genus Asperaceae.
(3) 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside according to (2), wherein the plant belonging to the genus Asperaceae is a hop (Humulus lupulus).
(4) An antioxidant comprising the compound according to (1) as an active ingredient.
(5) A pharmaceutical comprising the compound according to (1) as an active ingredient.
(6) A cosmetic comprising the compound according to (1) as an active ingredient.
(7) A food or drink comprising the compound according to (1) as an active ingredient.

  The pharmaceuticals, cosmetics, foods and drinks containing 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside or pharmaceutically acceptable salts thereof of the present invention are Since it has an oxidizing action, it has the effect of suppressing damage caused by active oxygen and lipid peroxide generated in the living body. Therefore, it is effective for the prevention and treatment of health disorders and cosmetic disorders, and it is useful for stabilizing foods and beverages and improving storage stability. It is industrially valuable.

  The present inventors obtained a novel 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D- represented by the above general formula [1] from hop (Humulus lupulus). Glucopyranoside was isolated and found to have strong antioxidant activity, thereby completing the present invention.

  As a method for isolating 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside from hop (Humulus lupulus), for example, it was described in WO 2004/052898 pamphlet. What is necessary is just to extract by a well-known method.

  When 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside of the present invention is used as a pharmaceutical, food, drink or cosmetic, it can be used in an advantageous form, It can also be used in the form of a pharmaceutically acceptable salt.

  Examples of the pharmaceutically acceptable salt include alkali metal salts such as sodium salt, potassium salt and lithium salt, alkaline earth metal salts such as calcium salt and magnesium salt, iron salt and copper salt. Examples include transition metal salts, ammonium salts, ethylamines, triethylamines, diethanolamine salts, triethanolamine salts, dicyclohexylamine salts, dimethylaminoethanol, arginine salts, ethylenediamine salts, and the like.

  These salts can be produced by conventional means, for example in the form of the advantageous phenolic acid in a solvent or solute in which the salt is insoluble or in a solvent such as water that can be removed by vacuum drying or freeze drying. The compound of the present invention can be produced by reacting the compound of the present invention with one equivalent or more of a suitable base, or by exchanging existing salt ions with other ions using a suitable ion exchange resin.

  Alternatively, 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside or a pharmaceutically acceptable salt thereof of the present invention is a commonly used carrier, auxiliary agent, It can be formulated together with additives and the like, and can be used in the fields of pharmaceuticals, quasi-drugs, cosmetics and the like as oral and parenteral products according to a standard method. Or as a food material, it can mix and can be made into food-drinks.

  Pharmaceutical products include tablets, capsules, granules, syrups and the like as oral preparations, and external preparations such as ointments, creams and liquids as parenteral preparations, and injections such as sterile solutions and suspensions. When these products are administered to a disease as a medicine, 2 mg to 500 mg can be administered once to several times a day, that is, a total daily dose of 2 mg to 1000 mg, and the effect can be sufficiently exerted.

  2- (2-Methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside or a pharmaceutically acceptable salt thereof of the present invention is a physiologically recognized vehicle, carrier, The required unit volume can be taken together with the form, binder, stabilizer, flavoring agent and the like. The adjuncts mixed in tablets and capsules are as follows. Binders such as tragacanth gum, gum arabic, starch, gelatin, excipients such as microcrystalline cellulose, starch, pregelatinized starch, swelling agents such as alginic acid, lubricants such as magnesium stearate, Sweeteners such as sugar, lactose, saccharin, flavoring agents such as peppermint, red mono oil, and cherry. In addition, the mixture of capsules can contain a liquid carrier such as fats and oils in addition to the above materials, and other materials can be used as coating agents or to change the physical form of the formulation in other ways. Can exist. For example, tablets can be coated with shellac, sugar. A syrup or elixir may contain the active compound, sucrose as a sweetening agent, methyl or propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor.

  Sterile compositions for injections dissolve or suspend active substances in vehicles such as water for injection, naturally occurring vegetable oils such as sesame oil, coconut oil, peanut oil, cottonseed oil, or synthetic fat vehicles such as ethyl oleate. It can be formulated by the usual method of turbidity. Alternatively, a buffer material, an antiseptic, an antioxidant and the like can be blended as necessary.

  As an external preparation, petrolatum, paraffin, oils and fats, lanolin, macrogol and the like are used as a base, and an ointment, a cream and the like are obtained by a usual method.

  The food and drink to which 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside of the present invention or a pharmaceutically acceptable salt thereof is added may be in the form of the above preparation. Although it is good, it is processed and manufactured by a general manufacturing method by adding a predetermined amount to each food ingredient in the form of candy, rice cracker, cookie, juice or the like. Ingestion as a health food or functional food is used for disease prevention and health maintenance, so it is taken as a processed product containing 5 mg to 500 mg as a total daily dose, divided into several times a day for oral intake.

Further, 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside of the present invention includes compounds having antioxidant activity such as vitamin C, vitamin E, and coenzyme Qn. Can be used together. Moreover, these substances have an effect | action also as a stabilizer of this food-drinks, and it is useful to use together. These antioxidants are added below the normal dose.
EXAMPLES The present invention will be described more specifically with reference to the following examples. However, the scope of the present invention is not limited to these examples.

2- (2-Methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside purified hop (Humulus lupulus) obtained from cocoon (hereinafter referred to as hop cocoon) known literature (WO2004) / 052898 pamphlet), a hop koji polyphenol fraction was prepared. That is, the obtained hop koji was extracted with 50% ethanol at 50 ° C. for 10 hours, the precipitate was removed by centrifugation, and the supernatant was concentrated under reduced pressure. The obtained extract was redissolved in water, applied to a column packed with synthetic resin SEPABEADS SP850 (Mitsubishi Chemical Corporation), washed with water, and eluted with 50% ethanol. This fraction was spray-dried, and the resulting brown powder was used as the hop straw polyphenol fraction. 150 g of hop koji polyphenol fraction was dissolved in 350 ml of distilled water, and the solvent was fractionated three times with 350 ml of hexane, and then the remaining aqueous layer was three times with 350 ml of diethyl ether. Next, the water fraction was pulverized by lyophilization under reduced pressure. The obtained powder was applied to a SEPABEADS SP850 resin column (Mitsubishi Chemical Corporation) and eluted stepwise with water, 25% ethanol, and 50% ethanol. The 25% ethanol fraction was concentrated under reduced pressure and lyophilized. 2.6 g of the residue was subjected to high performance liquid chromatography (HPLC) under the following conditions, and a peak at 60 minutes was separated and purified.
HPLC condition column: Inertsil ODS-3 (φ20 × 250 mm, GLsciences)
Flow rate: 10 ml / min
Mobile phase: A solution 0.1% formic acid aqueous solution, B solution acetonitrile: 0.1% formic acid aqueous solution = 1: 10 → 70 min: 90% A → 70% A
Detection: UV280nm

  Lyophilization gave 9.28 mg of 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside as a pale yellow powder.

The molecular formula of 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside and the structure of the purified compound were determined using TOF-MS (time-of-flight mass spectrometry). The structural formula was determined using NMR (nuclear magnetic resonance).

TOF-MS measurement "UPLC / TOF-MS conditions"
HPLC: ACQUITY UPLC (Waters)
Column: ACQUITY UPLC BEH C18 (50 × 2.1 MM, 1.7 μm: Waters)
Mobile phase: A liquid-methanol B liquid-0.1 (V / V)% formic acid aqueous solution
10 (V / V) A liquid (0 min) → 50 (V / V) A liquid (5 min)
Flow rate: 0.6 ml / min
Column temperature: 40 ° C
Injection volume: 5 μl
Detector: LCT Premier (Waters)
Ion source: ESI (positive, negative),
Cone voltage: 30V
Capillary voltage: 2500V
Ion source temperature: 120 ° C
Desolvation temperature: 300 ° C
MS scan: m / z 100-1000
The signal of [M + H] ⁺ = 521 was observed in the positive ion mode measurement, and the signal [M−H] ⁻ = 519 was observed in the negative ion mode measurement. Therefore, the integer molecular weight was determined to be 520. Moreover, the molecular formula was determined as C ₂₂ H ₃₂ 0 ₁₄ from the results of high-resolution mass spectrometry.

The compound purified by NMR measurement was dissolved in CD3OD. Various NMR spectra (1H, 13C, HSQC, HMBC) were measured using XWIN-NMR AV600 (Bruker Biospin) for NMR measurement. Table 1 shows the measurement results. As a result, the compound was determined to be 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside (hereinafter sometimes abbreviated as MPOG).

(Measurement of antioxidant activity)
DPPH radical scavenging activity

For the known acylphloroglucinol monoglycoside 2- (2-methylpropanoyl) phloroglucinol-1-O-β-D-glucopyranoside (Chemical Formula 2), a new compound MPOG, DPPH (1,1-diphenyl- (2-picrylhydrazyl) radical scavenging activity was measured. Specifically, a dilution series of samples was prepared in a 96-well plate, 100 μl of 1 well was added, 100 μl of 750 μM DPPH solution (50% methanol solution) was added, and the mixture was allowed to stand at room temperature for 30 minutes, and then the absorbance at 550 nm was measured. The final concentration (IC ₅₀ ) of the test substance that reduces the absorbance by half was determined and used as an index of antioxidant activity. The results are shown in FIG. MPOG showed stronger activity than 2- (2-methylpropanoyl) phloroglucinol-1-O-β-D-glucopyranoside. (IC ₅₀ are 0.10 mM and 1.01 mM, respectively).

  From these results, it was revealed that a novel acyl phloroglucinol disaccharide glycoside having a high antioxidant action could be obtained.

(Tablets and capsules)
10% of the compound (MPOG) obtained in Example 1
Lactose 75%
Magnesium stearate 15%
The above ingredients were uniformly mixed to obtain tablets and capsules.

(Powder, granule)
Compound obtained in Example 1 (MPOG) 20%
Starch 30%
Lactose 50%
The above ingredients were mixed uniformly to obtain a powder and granules.

(Injection)
Compound obtained in Example 1 (MPOG) 1%
Surfactant 9%
Saline 90%
The above ingredients were mixed by heating and sterilized to give an injection.

(cookie)
A wheat flour containing 2% by weight of the compound (MPOG) obtained in Example 1 and seasoned with salt, sucrose, butter, etc., is thoroughly stirred with an appropriate amount of water, baked at 190-200 ° C. for 30 minutes, and cookie. It was.

(jelly)
13 g of agar was dissolved by heating in 1 liter of water, 500 g of sucrose, 150 g of starch syrup and a little salt were added, and the mixture was heated and dissolved with stirring. Then, 2% by weight of the compound (MPOG) obtained in Example 1 and fruit juice Then, coloring and flavoring were added and cooled to make a jelly.

(Rain)
20 parts by weight of sucrose and 10 parts by weight of starch syrup (75% solids) were added and mixed with 10 parts of water. After heating and stirring at 150 ° C., 2% by weight of the compound (MPOG) obtained in Example 1 and coloring Add fragrance, fragrance, etc. and cool to make candy.

(Hand lotion)
15 parts by weight of carbon wax 1500, 8 parts by weight of alcohol and 90 parts by weight of propylene glycol were mixed and dissolved well, 52.5 parts by weight of water, 2 parts by weight of the compound (MPOG) obtained in Example 1, and a fragrance and preservative. An appropriate amount was added to obtain a hand lotion.

(External preparation)
Ethyl paraoxybenzoate 0.1%, butyl paraoxybenzoate 0.1%, lauromacrogol 0.5%, cetanol 18%, white petrolatum 40%, water 37.3%, the compound obtained in Example 1 ( MPOG) 4% was mixed to make an ointment according to a conventional method.

  According to the present invention, a new hop-derived compound having an antioxidant action can be provided. Since it has an antioxidant action, it has an effect of suppressing damage caused by active oxygen and lipid peroxide generated in the living body. Therefore, it is effective for the prevention and treatment of health disorders and cosmetic disorders, and can be applied and processed in medicines, foods and drinks, and cosmetics.

The graph which shows the result of a DDPH assay.

Claims (7)

Chemical formula [1]

2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside or a pharmaceutically acceptable salt thereof, wherein The 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside according to claim 1, which is obtained from a plant belonging to the genus Asperaceae. The 2- (2-methylpropanoyl) phloroglucinol-1,5-di-O-β-D-glucopyranoside according to claim 2, wherein the plant of the genus Asaaceae is a hop (Humulus lupulus). An antioxidant comprising the compound according to claim 1 as an active ingredient. A pharmaceutical comprising the compound according to claim 1 as an active ingredient. A cosmetic comprising the compound according to claim 1 as an active ingredient. A food or drink comprising the compound according to claim 1 as an active ingredient.

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