JP2016196413A - Monieroside a and derivative thereof, or pharmaceutical composition, health food or cosmetic comprising the same as active ingredient - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide Monieroside A and a specific derivative thereof, or useful materials comprising them, such as pharmaceutical compositions, health foods, food additives, and supplements.SOLUTION: Bioactivities of Monieroside A and a specific derivative thereof, such as acetylcholine esterase inhibitory activity and AGEs (advanced glycation end products) production inhibitory activity, are applied for the industry fields such as cosmetics, food additives, supplements, health foods, and pharmaceuticals.SELECTED DRAWING: Figure 1

Description

  The present invention relates to moniloeroside A and derivatives thereof, or a pharmaceutical composition, health food or cosmetic containing the same as an active ingredient.

  Bacopa monniera is a wet perennial widely distributed in Europe, North Africa, Asia, and the Americas. It is used as an ornamental aquatic plant in Japan and is one of the herbs used in traditional medicine (Ayurveda) in India. Otomeazena is said to have an effect (Patent Document 1) such as “Improving memory” and “Reducing anxiety”, and has been used for a long time as a tonic of the brain. Applications are also being studied (Patent Document 2). As the contained component, a dammarane type triterpene glycoside component, a cucurbitan type triterpene glycoside component, and the like have been reported.

JP 2005-73504 Gazette Japanese Patent Laid-Open No. 3-58939

  Regarding the pharmacological activity of otomezena, the extract has been reported to have neuroprotective action, antidepressant action, antioxidant action (Patent Document 1) and the like. For the first time, the present inventors isolated Monielloside A from an extract of whole Indian otameazena. As a result of further intensive studies, the present inventors have found that monielloside A has an acetylcholinesterase inhibitory action, is useful for the prevention and treatment of Alzheimer's disease, is useful as an appetite regulator, and is used for the prevention or treatment of aging. Obtained various unexpected new findings such as being useful, useful as cosmetics, and further useful as pharmaceutical compositions, health foods, food additives, supplements, etc. It came to complete.

  This monieroside A is considered to have some contribution to the various pharmacological activities of the above-mentioned otomeazena, but it is completely known about pharmaceutical compositions, cosmetics and cosmetics containing monieroside A itself and its derivatives as active ingredients. It is not done. That is, an object of the present invention is to provide monieroside A and a specific derivative thereof, and provide a useful pharmaceutical composition, health food, food additive, supplement and the like containing monieroside A and the specific derivative thereof. There is.

That is, the present invention relates to the following inventions.
[1] Structural formula (1)
Or a pharmacologically acceptable salt, ester or etherified product thereof.
[2] The compound according to [1], wherein the esterified product is acylated on one or more hydroxyl groups in the structural formula (1).
[3] The compound according to [1], wherein the etherified product is obtained by etherifying one or more hydroxyl groups in the structural formula (1).
[4] A pharmaceutical composition for preventing or treating a disease, comprising the compound according to any one of [1] to [3].
[5] The pharmaceutical composition according to [4], which is an acetylcholinesterase inhibitor.
[6] The pharmaceutical composition according to claim 4, wherein the disease is Alzheimer's disease.
[7] A food additive, supplement or health food comprising the compound according to any one of [1] to [3].
[8] The food additive, supplement or health food according to [7], which is an appetite regulator.
[9] A cosmetic comprising the compound according to any one of [1] to [3].
[10] The cosmetic product according to [9], which is an AGEs (advanced glycation end products: hereinafter also referred to as “terminal glycation product”) production inhibitor.

  ADVANTAGE OF THE INVENTION According to this invention, the pharmaceutical composition, health food, food additive, supplement, cosmetics etc. which have the outstanding physiological activity can be provided. In addition, monieroside A and its derivatives useful as raw materials for producing pharmaceutical compositions, health foods, food additives, supplements, cosmetics, and the like can be provided. The above-mentioned pharmaceutical composition, health food, food additive, supplement, cosmetics, etc., and a method for producing Monieroside A and its derivatives useful as production raw materials can be provided. The physiological activity includes acetylcholinesterase inhibitory action, appetite regulating action, AGEs production inhibitory action, and the like.

FIG. 5 is a diagram showing an outline of a process for determining the structure of monielloside A.

  Monielloside A is obtained from Otomeazena. Without being limited to otomezena, any plant containing monieroside A can be used as a production raw material. The place of origin may be from any place of origin, such as Europe, North Africa, Asia, and the Americas, but Indian Otome Azena is used for convenience.

  Monieroside A is solvent-extracted from the above plant, and Monielloside A is obtained from the extract and, if desired, is subjected to purification means and isolated. Otome Zena used for extraction may be whole plant, or a part thereof or a processed product thereof. Examples of the extract include flowers, fruits, leaves, stems, roots, and the like, and may be a cut product of the plant, but is preferably a dried product.

  The solvent used for the solvent extraction may be any solvent that dissolves monieroside A. Examples of excellent extraction solvents include water, alcohols such as methanol, ethanol, isopropanol, and n-butanol. For example, ethers such as dioxane, methyl ethyl ether, diethyl ether, etc., ketones such as acetone, methyl ethyl ketone, halogenated hydrocarbons such as dichloromethane, dichloroethane, etc., polar solvents such as dimethylformamide, dimethyl sulfoxide, etc. Etc. are exemplified.

  In the extraction operation, for example, the extract and the extraction solvent are mixed, preferably stirred, and the extract and the extraction residue are separated. Although the extraction temperature cannot be generally specified, it is preferably 15 ° C. to 150 ° C., and the extraction time cannot be generally specified, but is preferably about 30 minutes to 1 week. The separation of the extraction liquid and the extraction residue is not particularly limited, and is preferably performed by means such as filtration and centrifugation. By removing the extraction solvent from the extract, an otomezena extract can be obtained. The extraction solvent is removed under normal pressure or reduced pressure, preferably under reduced pressure.

Otomeazena extract contains monielloside A. Monieroside A can be obtained by subjecting the extract of otomezena to a purification means. Such purification means may be appropriately selected or combined, for example, from partition extraction, concentration, chromatography, crystallization, recrystallization and the like. Such purification means have been well developed in the art, and may be followed in the present invention. As a solvent combination for partition extraction, a combination of ethyl acetate and water or a combination of n-butanol and water is preferable. Chromatography includes normal phase silica gel, reverse phase ODS column chromatography, and reverse phase HPLC, but these may be repeated in appropriate combinations.
In this way, Monielloside A can be acquired, but confirmation of acquisition can be performed by, for example, optical rotation or MS spectrum.

  Examples of the salt of monieroside A include alkali metal salts such as sodium salt and potassium salt, and alkaline earth metal salts such as magnesium salt and calcium salt. The preparation of the salt will be obvious to those skilled in the art. The esters of monieroside A are obtained by esterifying the phenolic hydroxyl group of monieroside A with carboxylic acid, sulfonic acid, or phosphonic acid. The esterification may be carried out by a means known per se, for example, by reacting monieroside A with the acid chloride of the above acid. Examples of the ester residue include alkylcarbonyl, alkylsulfonyl, alkylphosphoryl, and the like. As the alkyl group, for example, C1-C4 lower alkyl such as methyl, ethyl, propyl, and butyl is preferable.

The ether of monielloside A can be obtained, for example, by reacting monielloside A or a pharmacologically acceptable salt thereof with a halide (for example, alkyl halide). Since the etherification reaction has been well established in the past, it can be followed in the present invention. As the alkyl group in this case, for example, C1-C4 lower alkyl such as methyl, ethyl, propyl, and butyl is preferable.
Acquisition of the esters or ethers from the reaction and reaction mixture may be carried out by means known per se, for example, by reacting Monielloside A with the halide, partition extraction, concentration, chromatography, crystallization, The target substance can be obtained by appropriate selection or combination from recrystallization and the like.

[Pharmaceutical]
The present invention provides a medicine containing monieroside A and its derivatives. Since monieroside A has acetylcholinesterase inhibitory activity, it can be used as a drug for regulating appetite or a drug for Alzheimer-type dementia. Moreover, since monieroside A has an AGEs production | generation inhibitory effect, it can be used as pharmaceuticals, such as aged skin defense and an internal use or external preparation for improvement.

  The medicament of the present invention comprises monieloside A and its derivatives as active ingredients, and a pharmaceutically acceptable carrier or additive according to a known method for producing a pharmaceutical preparation (for example, a method described in Japanese Pharmacopoeia). It can be formulated by mixing as appropriate. Specifically, for example, tablets (including sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets, buccal tablets, etc.), pills, powders, granules, capsules (including soft capsules and microcapsules) , Lozenges, syrups, solutions, emulsions, suspensions, controlled-release preparations (eg, immediate-release preparations, sustained-release preparations, sustained-release microcapsules), aerosols, films (eg, oral disintegration) Film, oral mucosa adhesive film), injection (eg, subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection), drip, transdermal preparation, ointment, lotion, patch Suppositories (eg, rectal suppositories, vaginal suppositories), pellets, nasal preparations, pulmonary preparations (inhalants), ophthalmic preparations, and oral or parenteral preparations. What is necessary is just to set suitably about the mixture ratio of a carrier or an additive based on the range normally employ | adopted in the pharmaceutical field | area. Carriers or additives that can be blended are not particularly limited. For example, various carriers such as water, physiological saline, other aqueous solvents, aqueous or oily bases; excipients, binders, pH adjusters, disintegrants, absorption Various additives such as an accelerator, a lubricant, a colorant, a corrigent, and a fragrance are included.

  Additives that can be mixed into tablets, capsules and the like include binders such as gelatin, corn starch, tragacanth, gum arabic, excipients such as crystalline cellulose, corn starch, gelatin, alginic acid and the like. Leavening agents, lubricants such as magnesium stearate, sweeteners such as sucrose, lactose or saccharin, flavorings such as peppermint, red oil and cherry. When the dispensing unit form is a capsule, a liquid carrier such as fats and oils can be further contained in the above type of material. Sterile compositions for injection can be prepared according to normal pharmaceutical practice (for example, dissolving or suspending an active ingredient in a solvent such as water for injection or natural vegetable oil). As an aqueous solution for injection, for example, isotonic solutions (eg, D-sorbitol, D-mannitol, sodium chloride, etc.) containing physiological saline, glucose and other adjuvants are used. For example, alcohol (eg, ethanol), polyalcohol (eg, propylene glycol, polyethylene glycol), nonionic surfactant (eg, polysorbate 80TM, HCO-50) and the like may be used in combination. As the oily liquid, for example, sesame oil, soybean oil and the like are used, and they may be used in combination with solubilizing agents such as benzyl benzoate and benzyl alcohol. Buffers (eg, phosphate buffer, sodium acetate buffer), soothing agents (eg, benzalkonium chloride, procaine, etc.), stabilizers (eg, human serum albumin, polyethylene glycol, etc.), storage You may mix | blend with an agent (for example, benzyl alcohol, phenol, etc.), antioxidant, etc.

  Since the preparation thus obtained is safe and has low toxicity, for example, it is administered to humans and other mammals (eg, rats, mice, rabbits, sheep, pigs, cows, cats, dogs, monkeys, etc.) can do.

  The medicament of the present invention varies depending on the dosage form, administration method, carrier, etc., but monieroside A or a derivative thereof is usually 0.01 to 100% (w / w), preferably 0.1 to 95%, based on the total amount of the preparation. It can manufacture in accordance with a conventional method by adding in the ratio of (w / w).

  The dose varies depending on the administration subject, symptom, administration route and the like, but in the case of oral administration, for example, generally about 0.01 to 1000 mg, preferably about 0.1, per day for a human body weight of about 60 kg. 1 to 100 mg, more preferably about 0.5 to 50 mg. The total daily dose may be a single dose or divided doses.

  The medicament of the present invention can be additively or synergistically used in combination with other medicaments for Alzheimer-type dementia, or other medicaments for protecting against aging skin, for improving internal use, or for external use. The improvement of the effect can be expected.

[Food and Drink]
This invention provides the food-drinks containing the monelloyside A and its derivative (s). The food / beverage product of the present invention is suitable as an appetite regulating food / beverage product because Monieroside A has acetylcholinesterase inhibitory activity, and Monielloside A is suitable as a food product for preventing aging skin and improving, since it has an AGEs production inhibitory action. The food and drink includes health food, functional food, food for specified health use, and food for the sick. The form of the food or drink is not particularly limited. For example, beverages such as tea beverages, soft drinks, carbonated beverages, nutritive beverages, fruit beverages, lactic acid beverages, buckwheat noodles, Chinese noodles, instant noodles and other noodles, rice cakes, candy, gum, chocolate, snacks, biscuits, jelly, jam , Cream, baked confectionery, confectionery such as bread, and fishery products such as kamaboko, ham, sausage, processed food, dairy products such as processed milk, fermented milk, salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream And fats and fat processed foods such as dressings, seasonings such as sauces and sauces, retort pouch foods such as curry, stew, rice cakes, rice cakes and miscellaneous foods, and frozen desserts such as ice cream, sherbet and shaved ice.

  The present invention provides supplements containing monieroside A and its derivatives. The supplement of the present invention is particularly suitable as a supplement for appetite regulation, or as a supplement for preventing and improving aging skin. Supplements can be provided in the form of tablets, granules, powders, drinks, and the like.

  The present invention provides food additives containing monielloside A and its derivatives. The food additive of the present invention is suitable as a food additive for regulating appetite, or a food additive for preventing and improving aging skin. The form of the food additive of the present invention is not particularly limited, and examples thereof include a liquid form, a paste form, a powder form, a flake form, and a granular form. The food additive of this invention can be manufactured according to the manufacturing method of a general food additive. Furthermore, it can also be implemented as a feed or feed additive containing monielloside A and its derivatives.

  The present invention provides a cosmetic containing Monielloside A and its derivatives. The cosmetic product of the present invention is particularly suitable as a cosmetic product for protecting and improving aging skin. Cosmetics include so-called medicinal cosmetics (quasi-drugs). Cosmetics include, for example, cleaning agents, shampoos, rinses, hair nicks, hair lotions, after shave lotions, body lotions, cosmetic lotions, cleansing creams, massage creams, emollient creams, aerosol products, deodorants, fragrances, deodorants or Examples include bath salts. The cosmetic product of the present invention is a component commonly used as a cosmetic product other than the lecithin of the present invention, for example, surfactants, moisturizers, animal and plant derived oils and fats, silicones, higher alcohols, lower alcohols, animal and plant derived extracts, ultraviolet absorption Agents, flame retardants, metal sequestering agents, vitamins, antioxidants, thickeners, preservatives, bactericides, pH adjusters, colorants, various fragrances and the like can be appropriately blended depending on the purpose.

<Extraction and purification operations>
A dried whole plant (2.0 kg) of Indian Otomeazena (Bacopa monniera) was extracted with methanol when heated (80 ° C.). The extract was filtered, methanol was added to the residue, and the same extraction operation was performed 3 times. The methanol extracts were combined, and the solvent was distilled off under reduced pressure to obtain a methanol extract (608.82 g, yield 30.4%).

The methanol extract (608.82 g) was subjected to solvent partitioning with ethyl acetate, n-butanol, and water. As a result, the ethyl acetate fraction (31.8 g, yield 4.8%), the n-butanol fraction (57.94 g, yield 8.8%), and the water fraction (110.3 g, yield 16.). 8%).
Thus, the ethyl acetate fraction was repeatedly separated and purified by normal phase silica gel, reverse phase ODS column chromatography and reverse phase HPLC, and as a result, a new compound, Monieroside A was obtained.
These compounds were identified by ¹ H-NMR and ¹³ C-NMR spectrum data, optical rotation and MS spectrum.

<Structural analysis of Monielloside A>
Monieroside A was obtained as an amorphous powder exhibiting negative optical rotation ([α] _D ²⁷ -3.9 °, methanol). The IR spectrum suggested the presence of hydroxyl group (3415 cm ⁻¹ ), ester (1705 cm ⁻¹ ) and aromatic (1609 cm ⁻¹ ). Furthermore, a pseudo-molecular ion peak was observed at m / z 473 (M + Na) ⁺ in positive-ion FAB-MS, and it was found from high resolution FAB-MS that the compound had the molecular formula C ₂₁ H ₂₂ O ₁₁ .

Monieroside A was hydrolyzed with 5% by weight aqueous potassium hydroxide to obtain p-hydroxybenzoic acid and identified by HPLC. Monieroside A was further added with 5% by weight hydrous sulfuric acid-1,4-dioxane. When acid hydrolysis was performed with (1: 1), D-glucose was confirmed. Identification including the absolute configuration of the constituent sugars was performed by derivatizing into tolylthiocarbamoyl thiazolidine and analyzing by HPLC [Tanaka T., Nakashima T., Ueda T., Tomii K., Kouno I., Chem. Pharm. Bull., 55, 899-901 (2007)]. From the HMBC correlation, it was confirmed that glucose was bonded to the 8th position of aglycone and p-hydroxybenzoic acid was bonded to the 2nd position thereof.
From the above results, the above chemical structure [Structural Formula (1)] of Monielloside A was determined.

<Evaluation of acetylcholinesterase inhibitory activity>
Acetylcholinesterase inhibitors are generally used as symptomatic drugs for Alzheimer's symptoms, but the main side effects of cholinesterase inhibitors in the center are common symptoms of digestive system such as anorexia, nausea and vomiting, Drugs that improve anorexia by promoting the secretion of acetylcholine, which is less secreted, and increasing gastrointestinal motility in the vicinity of the gastrointestinal tract. It is also known that the appetite can be controlled by adjusting the amount of acetylcholine depending on the patient's condition.

When the inhibitory action of acetylcholinesterase was examined for monieroside A, the results shown in Table 1 were obtained, and a significant inhibitory action was observed. This suggests that monielloside A significantly regulates appetite, and supports its usefulness as a medicine for regulating appetite. Moreover, the usefulness as said Alzheimer therapeutic agent is also supported from this effect | action.

<AGEs production inhibitory action>
In recent years, new cosmetic materials have been developed for the purpose of “anti-aging”, and in particular, AGEs, which are reaction products of protein saccharification reactions, are attracting attention as substances involved in aging. The search for the substance that suppresses is widely conducted. In this study, we examined the AGEs production inhibitory action of the extract of otomeazena and the new ingredient monielloside A. The specific experimental procedure is as follows.

  100 μL of BSA solution [bovine serum albumin (manufactured by Nacalai Tesque) 20 mg / mL, sodium azide (manufactured by Sigma Ardrich) 2 mg / mL], 80 μL of D-glucose solution (concentration: 250 mg / mL) and 20 μL of the prepared sample were mixed. . This was reacted at 60 ° C. for 72 hours, then transferred to a 96-well micro black plate (Nunc), and the fluorescence intensity was measured (Ex: 355 nm, Em: 440 nm). After adding the sample to the reaction product prepared at the same time, the fluorescence intensity was measured, the fluorescence quenching ability of the sample was also measured, and the reaction inhibition rate was corrected. Table 2 shows the AGEs inhibitory activity of the methanol extract and each fraction, and Table 3 shows the AGEs inhibitory activity of Monielloside A.

On the other hand, when a similar test was carried out using aminoguanidine hydrochloride, which is widely known as a substance that suppresses the generation of AGEs, as a control substance, the production inhibition rate at 100 μg / mL under the same experimental conditions was 29%. From this, it was found that the methanol extract of otomezena, each extract, and the new component monelloyside A can strongly suppress the generation of AGEs and contribute to the suppression of various in vivo aging phenomena.

  Monieroside A and its derivatives, obtained from Bacopa monniera, have physiological activities such as acetylcholinesterase inhibitory activity and AGEs production inhibitory activity, so they are used in cosmetics, food additives, supplements, health foods, pharmaceutical fields, etc. Can be used in a wide range of fields.

Claims (10)

The following structural formula (1)
Or a pharmacologically acceptable salt, ester or etherified product thereof.   The compound according to claim 1, wherein one or two or more hydroxyl groups in the structural formula (1) are acylated in the esterified product.   2. The compound according to claim 1, wherein the etherified product is obtained by etherifying one or more hydroxyl groups in the structural formula (1).   A pharmaceutical composition for preventing or treating a disease, comprising the compound according to any one of claims 1 to 3.   The pharmaceutical composition according to claim 4, which is an acetylcholinesterase inhibitor.   The pharmaceutical composition according to claim 4, wherein the disease is Alzheimer's disease.   A food additive, supplement or health food comprising the compound according to any one of claims 1 to 3.   The food additive, supplement or health food according to claim 7, which is an appetite regulator.   A cosmetic comprising the compound according to any one of claims 1 to 3.   The cosmetic according to claim 9, which is an AGEs production inhibitor.