JP2009196902A - New saponin compound containing ilex paraguayensis and its use - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To develop a novel use of Ilex paraguayensis. <P>SOLUTION: An extract of Ilex paraguayensis that can be used safely is provided. A free fatty acid production inhibitor and an antiobesitic or antihyperlipidemic composition comprising as an effective ingredient at least one of saponin compounds contained in the extract are provided. A health food comprising the composition is also provided. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

The present invention relates to a matecha extract and a novel saponin compound contained in the extract.
More specifically, the present invention relates to a water- or water-containing lower alcohol extract of dried leaves of matea, which is a genus Ilexaceae, and / or a novel saponin compound contained in the extract and use thereof.

  More specifically, the present invention comprises the above-mentioned extract or the extract, and a novel fatty acid production inhibitory action of a novel saponin compound contained in the extract, a pharmaceutical composition having the action, and the composition. Related to health food.

Aquifoliaceae plant Ilex paraguaiensis is an evergreen tree native to the mountains from southern Brazil to northeastern Argentina. The material is also referred to as material and paraguayan tree.
Since ancient times, this material has been used as drinking tea (mate tea) by drying the leaves with hot air and crushing them in South America (for example, Non-Patent Document 1).

  In addition, the material has been used in the manufacture of herbal medicines as a central nervous stimulant, diuretic and anti-rheumatic agent in South America since ancient times. From this material, uronic acid-3-O- [β -D-glucopyranosyl- (1 → 3) -α-L-arabinopyranosyl]-(28 → 1) -β-D-glucopyranosyl ester matesaponin 1 having an ester skeleton has been found (for example, non-patent Reference 2).

Furthermore, it is known that mate tea contains calcium, iron, anhydrous caffeine, tannin, vitamin A, vitamin B ₁ and vitamin B ₂ (for example, Patent Documents 1 and 2).
Mate tea also has tonicity, irritation, diuretic action; calms the nerves and raises the mind; works to compensate for malnutrition, stops weakness and promotes recovery; and helps digestion by promoting peristaltic movement of the intestines; It is known to have an effect of promoting respiratory exercise; an effect of enhancing the amount of labor of muscles; an effect of supplementing vitamin C; an effect of eliminating active oxygen and an effect of suppressing the onset of allergy (for example, Patent Documents 1 and 2).

  However, it has never been reported that the material has a lipase inhibitory activity effective for hyperlipidemia, obesity, or metabolic syndrome.

Incidentally, lipase is known to be an enzyme that hydrolyzes fat (triglycerides) contained in diets into free fatty acids and monoglycerides that can be absorbed by the body.
Thus, by inhibiting this lipase activity, the non-hydrolyzed triglyceride is excreted without being absorbed into the body, and as a result, it is possible to reduce fat intake, that is, reduce calorie intake, diabetes, It is expected to be effective in reducing calorie or fat intake in humans corresponding to hyperlipidemia and metabolic syndrome.

JP-A-6-135848 JP 2004-248601 A Edited by Mitsuru Hotta et al., World useful plant dictionary, page 549 (1989) Grace Gosmann et al., Journal of Natural Producets, 52 (6), pp. 1367-1370.

  An object of the present invention is to develop a new use of the material.

The present inventor started research to clarify the novel use of the matecha, and diligently researched the components contained in the pharmacological action of the extract of the matecha water or water-containing lower alcohol and the mateture extract at each purification stage, A new saponin compound was found.
The present inventor has also found that the extract of matecha and a novel saponin contained in the extract have lipase inhibitory activity.
Therefore, according to the present invention, the chemical structure of a novel saponin compound is determined, knowledge is also obtained about the pharmacological action of the compound, a novel use of the material is established, and the above-mentioned problems are solved.

［式中、
Ｒ¹は、α−Ｌ−アラビノピラノシル基、β−Ｄ−グルコピラノシル−(１→３)−α−Ｌ−アラビノピラノシル基またはα−Ｌ−ラムノピラノシル−(１→２)−Ｏ−[β−Ｄ−グルコピラノシル−(１→３)]−α−Ｌ−アラビノピラノシル基であり、Ｒ²は、β−Ｄ−グルコピラノシル基またはβ−Ｄ−グルコピラノシル−(１→６)−β−Ｄ−グルコピラノシル基であり、Ｒ³およびＲ⁴は、それぞれ独立して水素原子またはメチル基であり、Ｒ⁵はメチル基またはヒドロキシメチル基である］
で表されるサポニン化合物が提供される。 That is, according to the present invention, the following general formula (1):

[Where:
R ¹ is an α-L-arabinopyranosyl group, β-D-glucopyranosyl- (1 → 3) -α-L-arabinopyranosyl group or α-L-rhamnopyranosyl- (1 → 2)- O- [β-D-glucopyranosyl- (1 → 3)]-α-L-arabinopyranosyl group, R ² is β-D-glucopyranosyl group or β-D-glucopyranosyl- (1 → 6) ) -Β-D-glucopyranosyl group, R ³ and R ⁴ are each independently a hydrogen atom or a methyl group, and R ⁵ is a methyl group or a hydroxymethyl group.
The saponin compound represented by these is provided.

  In addition, the matecha water or water-containing lower alcohol extract and the matecha extract at each purification stage have a lipase inhibitory activity.

  Therefore, the material extract or composition containing the saponin compound according to the present invention can suppress the absorption of triglyceride by its lipase inhibitory activity, and is a pharmaceutical composition intended for anti-obesity, anti-hyperlipidemia and fatty liver improvement or It is used for health foods to which the composition is added.

  In addition, the material extract according to the present invention and the novel saponin compound contained in the extract can be safely used because they are obtained from the dried leaf portion of the material that has been used as a favorite product since ancient times.

  The present invention relates to a water or water-containing lower alcohol extract of a material containing a saponin compound represented by the above general formula (1) and / or the use of a material extract obtained by further purifying the extract as necessary.

The production area of matecha (Ilex paraguaiensis) used in the present invention may be any of the production areas of Southeast Brazil, Paraguay, Uruguay and Argentina where matecha grows naturally. It can be used without limitation.
In the present invention, the leaf portion of the texture is usually dried, but the collected leaf can be used as it is or after being made by a method known to those skilled in the art.

Regarding the solvent used for the preparation of the extract of the present invention, examples of the lower alcohol other than water include C1-C4 monoalcohols or dialcohols.
Specific examples include methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, t-butanol, a mixed solution thereof, or a hydrous alcohol in any ratio thereof. Furthermore, ethanediol, propanediol, butanediol, and their positional isomers or mixtures thereof, or hydrous alcohols in any proportions thereof.

However, from the viewpoint of ease of concentration after extraction, water, methanol or ethanol is preferably used. More preferably, water or about 80% ethanol is used, but in the present invention, the amount of ethanol in water-containing ethanol is not particularly limited to 80%.
These extraction solvents are used in an amount of about 1 to 50 times (volume), preferably about 2 to 10 times (volume) with respect to the extraction material.

  The extraction can be performed at room temperature or hot, and the extraction temperature can be arbitrarily set between room temperature and the boiling point of the solvent. In the case of hot extraction, for example, by immersing the dried leaf part of the texture in the extraction solvent at a temperature between 50 ° C. and the boiling point of the extraction solvent under shaking (or stirring) or non-shaking or refluxing. Is appropriate. When the extraction material is immersed under shaking, it is appropriate to carry out for about 30 minutes to 5 hours, and when it is immersed under non-shaking, it is appropriate to carry out for about 1 hour to 20 days. Moreover, when extracting under reflux of an extraction solvent, it is preferable to heat to reflux for 30 minutes to several hours.

  It is also possible to extract by dipping at a temperature lower than 50 ° C. In this case, it is preferable to immerse for a longer time than the above time. Although extraction operation may be performed only once about the same material, it is preferable from the point of extraction efficiency to repeat several times, for example, about 2 to 5 times.

The extract obtained by filtering the solid after extraction may be concentrated by an ordinary method to obtain an extract. Concentration is preferably performed under reduced pressure. Concentration may be performed until the extract is dry.
The extract may be used as it is to prepare the composition of the present invention, but may also be used as a powder or lyophilized product. A method known in the art can be adopted as the method for forming these solids.

  Therefore, the extract in the present invention means any of an extract, an extract, a concentrated dried product or a lyophilized product, but the extract according to the present invention can be used as it is without being purified. Part of.

  However, in the present invention, the extract obtained by concentrating the extract is subjected to partition extraction with a solvent, that is, partition extraction using water and a non-hydratable organic solvent one or more times, and the organic solvent-soluble fraction is extracted. And water-soluble fraction.

Examples of the non-hydratable organic solvent include ethyl acetate, n-butanol, hexane, chloroform and the like, and among them, ethyl acetate is preferable.
That is, the extract obtained by concentrating matecha water or water-containing lower alcohol extract is distributed using ethyl acetate and water as necessary to obtain an ethyl acetate-soluble fraction and a water-soluble fraction. Can do.

In addition, the water-soluble fraction obtained above can be further subjected to partition extraction using water and a non-hydratable organic solvent to separate the organic solvent-soluble fraction and the water-soluble fraction.
In this case, examples of the non-hydratable organic solvent include n-butanol, hexane, chloroform, and the like. Among them, n-butanol is preferable.
That is, the water-soluble fraction after partitioning between ethyl acetate and water is directly subjected to partitioning with n-butanol, or the residue obtained by concentrating the water-soluble fraction is further mixed with water and n-butanol. N-butanol fraction and water-soluble fraction can be obtained.

  The partition extraction can be performed according to a conventional method such as a stirring or shaking distribution method or a droplet counter-current distribution method usually performed in the art. For example, it is performed by adding about 1 to 10 times (volume) (1:10 to 10: 1) of a non-hydratable organic solvent and water to an extract or the like at room temperature, with shaking or without shaking. Is appropriate.

  Therefore, the material extract used in the present invention refers to the water extract of the material or the concentrate thereof, the ethyl acetate-soluble fraction or the concentrate thereof, or the n-butyl soluble fraction or the concentration thereof. Includes any of the objects.

The above water or lower alcohol extract and each partition extract can be subjected to purification treatment before and after concentration in any of the above steps.
For the purification treatment, in addition to the partition extraction of the water extract with water and a non-hydratable organic solvent, a chromatographic method, an ion exchange chromatographic method, etc. known to those skilled in the art may be used alone or in combination. Can do. For example, as a chromatographic method, a column chromatography, a thin layer chromatography, a high performance liquid chromatography, a centrifugal liquid chromatography or the like using a normal phase or reverse phase carrier or an ion exchange resin, or a combination thereof is performed. Is mentioned. In this case, purification conditions such as a carrier and an elution solvent can be appropriately selected according to various chromatographies.

  The present inventor examined the action of the purified product at the same time as the material extract was purified by combining the above purification methods.

  Therefore, the present inventor repeatedly performed chromatography and HPLC purification using a reverse phase column on the ethyl acetate fraction and the n-butanol fraction to isolate the contained components.

As a result, the present inventor obtained 10 kinds of known triterpene glycosides matesaponin 1, matesaponin 2, matesaponin 3, matesaponin 5, matesaponin J3a, matesaponin J3b, 3-O-β-D- from the ethyl acetate fraction. Glucopyranosyl (1 → 3) -α-L-2-O-acetylarabinopyranosyl ursolic acid 28-O-β-D-glucopyranoside, 3-O-α-L-rhamnopyranosyl (1 → 2) -α- L-arabinopyranosyl oleanolic acid 28-O-β-D-glucopyranosyl (1 → 6) -β-D-glucopyranoside, nudicaucin C, two known flavonoid glycosides rutin, kaempferol-3-O- Rutinoside, four known chlorogenic acids 3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid, 5-O-caffeoylquinic acid, 3,5-O-dicaffeoylquinic acid and cuff Give linalyl 6-O-α-L- arabinofuranosyl pyranosyl-beta-D-glucopyranoside as a known substance - in, theobromine and (R).
In addition, the present inventor has obtained from the n-butanol fraction one known triterpene glycoside matesaponin, one known triterpene, ursolic acid, 3-β-hydroxyurs-11-ene-13β (28)- In addition to obtaining orido, two known flavonoid glycosides rutin and kaempferol-3-O-rutinoside as known substances, the following novel compounds were also found.

［式中、
Ｒ¹は、α−Ｌ−アラビノピラノシル基、β−Ｄ−グルコピラノシル−(１→３)−α−Ｌ−アラビノピラノシル基またはα−Ｌ−ラムノピラノシル−(１→２)−Ｏ−[β−Ｄ−グルコピラノシル−(１→３)]−α−Ｌ−アラビノピラノシル基であり、
Ｒ²は、β−Ｄ−グルコピラノシル基またはβ−Ｄ−グルコピラノシル−(１→６)−β−Ｄ−グルコピラノシル基であり、
Ｒ³およびＲ⁴は、それぞれ独立して水素原子またはメチル基であり、
Ｒ⁵はメチル基またはヒドロキシメチル基である］
で表されるサポニン化合物が提供される。 That is, according to the present invention, the following general formula (1):

[Where:
R ¹ is an α-L-arabinopyranosyl group, β-D-glucopyranosyl- (1 → 3) -α-L-arabinopyranosyl group or α-L-rhamnopyranosyl- (1 → 2)- O- [β-D-glucopyranosyl- (1 → 3)]-α-L-arabinopyranosyl group,
R ² is a β-D-glucopyranosyl group or β-D-glucopyranosyl- (1 → 6) -β-D-glucopyranosyl group,
R ³ and R ⁴ are each independently a hydrogen atom or a methyl group,
R ⁵ is a methyl group or a hydroxymethyl group]
The saponin compound represented by these is provided.

で表わされるマテノシドＡが提供される。 In particular, in the compound of the above formula (1), in the compound of the above formula (1), R ¹ is a β-D-glucopyranosyl- (1 → 3) -β-D-arabinopyranosyl group, Formula (2), wherein R ² is a β-D-glucopyranosyl group, R ³ is a methyl group, R ⁴ is a hydrogen atom, and R ⁵ is a hydroxymethyl group:

Matenoside A represented by is provided.

で表わされるマテノシドＢが提供される。 According to the invention, in the compound of formula (1), R ¹ is an α-L-arabinopyranosyl group, R ² is a β-D-glucopyranosyl group, and R ³ is a methyl group. R ⁴ is a hydrogen atom, R ⁵ is a hydroxymethyl group, and the formula (3):

The matenoside B represented by is provided.

で表わされるマテノシドＣも提供される。 According to the invention, in the compound of formula (1), R ¹ is α-L-rhamnopyranosyl- (1 → 2) -O- [β-D-glucopyranosyl- (1 → 3)]-α-L. -Arabinopyranosyl group, R ² is β-D-glucopyranosyl- (1 → 6) -β-D-glucopyranosyl group, R ³ is a hydrogen atom, R ⁴ is a methyl group, R ⁵ is a methyl group, and the formula (4):

Matenoside C represented by is also provided.

Preparation of mate tea extract and isolation and purification of the extract-containing components For example, dry leaf (3.0 kg) of Brazilian mate (Ilex paraguariensis) was prepared using methanol (MeOH), and the mate extract was prepared, Examples of purification of the extract are shown in Table 1 and below.

(1) Preparation of matecha extract The dried leaf part (3.0 kg) of Brazilian matecha (Ilex paraguariensis) was extracted with MeOH (30 L) while hot and filtered, and the residue was subjected to the same operation twice. The solvent was distilled off under reduced pressure to obtain a MeOH extract (1.0 kg, yield 33.3%). 500.0 g of the obtained MeOH extract was suspended in H ₂ O ( ₂ L) and extracted three times with EtOAc (2 L). The H ₂ O soluble fraction was then extracted three times with n-BuOH (2 L), and each soluble fraction was treated according to a conventional method to obtain an EtOAc soluble fraction (168.3 g, 11.2%), n-BuOH. A soluble fraction (251.1 g, 16.7%) and an H ₂ O soluble fraction (96.5 g, 6.4%) were obtained.
The search for the components contained in the EtOAc-soluble fraction and n-BuOH-soluble fraction in which activity was observed was performed according to the procedure shown below.

(2) Purification of ethyl acetate (EtOAc) soluble fraction The EtOAc soluble fraction was repeatedly separated and purified by normal phase silica gel, reverse phase ODS column chromatography and HPLC to produce one novel triterpene glycoside matenoside B (0.0013). %), One known triterpene glycoside matesaponin 1 (0.010%), two known triterpene ursolic acids (1.02%), 3-β-hydroxyurs-11-ene-13β (28) -olide (0.039%) ) Two known flavonoid glycoside rutins (0.012%), kaempferol-3-O-rutinoside (0.0079%) were isolated.

(3) Purification of n-BuOH soluble fraction The n-BuOH soluble fraction obtained above was repeatedly separated and purified by normal phase silica gel, reverse phase ODS column chromatography and HPLC, and two new saponin compounds, matenoside. A (0.020%), matenoside C (0.20%), 10 known triterpene glycosides matesaponin 1 (0.080%), matesaponin 2 (0.029%), matesaponin 3 (0.0099%), matesaponin 4 (0.034%), matesaponin 5 (0.0094%), matesaponin J3a (0.10%), matesaponin J3b (0.060%), 3-O-β-D-glucopyranosyl (1 → 3) -α-L-2-O-acetylarabinopyranosyl ursol Acid 28-O-β-D-glucopyranoside (0.047%), 3-O-α-L-rhamnopyranosyl (1 → 2) -α-L-arabinopyranosyl oleanolic acid 28-O-β-D-glucopyranosyl (1 → 6) -β-D-glucopyranoside (0.29%), Nudicaucin C (0.12%), two known flavonoid glycosides rutin (0.50%), kaempferol-3-O-rutinoside (0.17%), four known chlorogenic acids 3-O-caffeoylquinic acid (0.16) %) 4-O-caffeoylquinic acid (0.15%), 5-O-caffeoylquinic acid (0.046%), 3,5-O-caffeoylquinic acid (0.32%) and caffeine (0.55%) ), Theobromine (0.15%), (R) -linalyl 6-O-α-L-arabinopyranosyl-β-D-glucopyranoside (0.090%) was isolated.

All of the above known compounds were identified by comparing ¹ H-NMR and ¹³ C-NMR spectrum data of these compounds with literature values.
However, the yield of these isolated components is the isolated yield from Brazilian material.

(4) Structural analysis of matenoside A and B Matenoside A was obtained as a white powder exhibiting positive optical rotation ([α] _D ²⁵ + 18.7 °, MeOH). The IR spectrum suggested the presence of hydroxyl group (3600 cm ^-1 ), carbonyl group (1716 cm ^-1 ), olefin (1650 cm ^-1 ), and ether (1070 cm ^-1 ).
Furthermore, pseudo-molecular ion peaks were observed at c / 951 951 (M + Na) ⁺ and 927 (M−H) ⁻ in the cation and anion FAB-MS, and the molecular formula C ₄₇ H ₇₆ O ₁₈ was observed from the high-resolution FAB-MS. It was found to be a compound having

By acid hydrolysis of matenoside A with 5% H ₂ SO ₄ aqueous solution-1,4-dioxane (1: 1, v / v, 1 ml), the known compound 23-hydroxyursolic acid is obtained, and as a constituent sugar L-arabinose and D-glucose were detected in the optical rotation detector.
Further, ¹ H-NMR and ¹³ C-NMR spectrum data (Tables 2 and 3) using pyridine-d _{5 of} matenoside A as a solvent and various two-dimensional NMR data were analyzed in detail.

The structure analysis of matenoside B was also conducted by the same method as described above. Matenoside B is obtained by acid hydrolysis with 5% H ₂ SO ₄ aqueous solution-1,4-dioxane (1: 1, v / v, 1 ml) to give the known compound 23-hydroxyursolic acid in the same manner as Matenoside A. As a constituent sugar, L-arabinose and D-glucose were detected by an optical rotation detector.
Further, ¹ H-NMR and ¹³ C-NMR spectrum data (Tables 1 and 2) of matenoside B using pyridine-d ₅ as a solvent and various two-dimensional NMR data were analyzed in detail.
From the above results, the chemical structures of matenosides A and B were determined as shown in the above formulas (2) and (3).

A schematic diagram of the above structure determination technique is shown below.

(4) Structural analysis of matenoside C The structural analysis of matenoside C was also carried out by the same method. Matenoside C was obtained by acid hydrolysis with 5% H ₂ SO ₄ aqueous solution-1,4-dioxane (1: 1, v / v, 1 ml) to obtain a known compound oleanolic acid and optical rotation as a constituent sugar. L-arabinose, D-glucose and L-rhamnose were detected in the detector.

Further, ¹ H-NMR and ¹³ C-NMR spectrum data (Tables 2 and 3) using pyridine-d _{5 of} matenoside C as a solvent and various two-dimensional NMR data were analyzed in detail.
From the above results, the chemical structure of matenoside C was determined as shown in the formula (4).

A schematic diagram of the above structure determination technique is shown below.

The present inventor has found that matecha extract, ie, methanol extract, ethyl acetate fraction, n-butanol fraction and water fraction, and matenoside C all have lipase inhibitory activity.
Further, the present inventor has also found that the known substances metasaponin 1 and nudicaucin C isolated from matecha extract have a lipase inhibitory activity.

  Further, the present inventor releases the material extract according to the present invention, that is, the methanol extract, the ethyl acetate fraction, the n-butanol fraction and the water fraction, and the matenoside C, based on the lipase inhibitory activity described above. The present invention was completed by discovering that it can be used for a pharmaceutical composition intended to suppress fatty acid production, anti-obesity, anti-hyperlipidemia, or a fatty liver ameliorating effect and a health food containing the composition.

  Therefore, according to the present invention, a free fatty acid based on a lipase inhibitory activity that contains a matecha extract, that is, a methanol extract, an ethyl acetate fraction, an n-butanol fraction, a water fraction, and / or matenoside C as an active ingredient. Production inhibition, and thus anti-obesity compositions are provided.

Further, according to the present invention, there is provided a composition for antihyperlipidemia comprising a extract of mate, ie, a methanol extract, an ethyl acetate fraction, an n-butanol fraction and a water fraction, and matenoside C as active ingredients. Is done.
In addition, according to the present invention, there is provided a composition for improving fatty liver comprising a extract of mate, ie, a methanol extract, an ethyl acetate fraction, an n-butanol fraction and a water fraction, and matenoside C as active ingredients. .

Therefore, according to the present invention, a high-calorie diet resulting from Westernization of dietary life, which includes a material extract, ie, a methanol extract, an ethyl acetate fraction, an n-butanol fraction and a water fraction, and matenoside C as active ingredients. A pharmaceutical composition intended for the prevention or treatment of lifestyle-related diseases such as obesity, hyperlipidemia, and fatty liver, which are representative of modern diseases based on the intake of food, and prevention or treatment of intestinal obstruction and constipation due to the promotion of intestinal motility Intended pharmaceutical compositions are provided.
Moreover, according to the present invention, a health food containing the composition is provided.

  The material extract of the present invention, that is, the methanol extract, the ethyl acetate fraction, the n-butanol fraction and the water fraction, or the concentrated dry solids thereof, or the matenoside C contained in the extract is in an intact state, or It can be used after being diluted with an appropriate medium or formulated into various pharmaceutical forms such as powders, granules, tablets, capsules or liquids by methods known in the field of pharmaceutical production.

  In these pharmaceutical forms, an appropriate medium may be added. Such vehicles include pharmaceutically acceptable excipients such as binders (e.g. syrup, gum arabic, gelatin, sorbitol, tragacanth or polyvinylpyrrolidone), fillers (e.g. lactose, sugar, corn starch, calcium phosphate, Sorbitol or glycine), lubricants (eg magnesium stearate, talc or polyethylene glycol), disintegrants (eg potato starch) or wetting agents (eg sodium lauryl sulfate).

  Tablets may be coated by conventional methods. Liquid formulations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, and may be provided as a dry product that is regenerated with water or other suitable excipients prior to use. Good.

  Such liquid preparations are made up of conventional additives such as suspending agents (e.g. sorbitol, syrup, methylcellulose, glucose syrup, gelatin edible fat), emulsifiers (e.g. lecithin, sorbitan monooleate or gum arabic), (edible fats). Non-aqueous excipients (e.g. almond oil, fractionated coconut oil or oily esters such as glycerin, propylene glycol or ethyl alcohol), preservatives (e.g. methyl or propyl p-hydroxybenzoate, or Sorbic acid) and, if desired, colorants or fragrances.

  Moreover, what added the composition by this invention to food as an active ingredient can be utilized as health food.

Health food means food with a more positive meaning than normal food for the purpose of health, health maintenance and promotion, for example, solid, semi-solid or liquid products, specifically powders, granules In addition to pills, tablets, capsules, liquids, and the like, confectionery such as cookies, rice crackers, jelly, yokan, yogurt, and manju, soft drinks, teas, nutritional drinks, soups, drinks, and the like.
In the production process of these foods or to the final product, the above-mentioned extract can be added to make a health food.

  The amount of matecha extract, that is, methanol extract, ethyl acetate fraction, n-butanol fraction and water fraction, and the amount of the compound according to the present invention varies depending on the age, symptoms, etc. When the product is used for prevention / treatment, it can be used at a dose of about 50 mg to 10 g, preferably about 100 mg to 3 g per adult. Moreover, when using as a health food, the said dry-solid thing can be used in the quantity which does not have a bad influence on the taste and external appearance of a foodstuff, for example, the said dried solid substance with respect to 1 kg of target foodstuffs.

  Examples of the material extract of the present invention, methods for purifying matenosides A, B, and C represented by formulas (2) to (4) and their actions will be specifically described below. It is for explaining the present invention and does not limit the present invention.

In the examples, unless otherwise specified, the following various solvents, chromatographic carriers, HPLC columns, and various analyzers were used:
Methanol (MeOH): Nacalai Tesque, special grade Ethanol (EtOH): Nacalai Tesque, special grade Chloroform: Nacalai Tesque, special grade Ethyl acetate (AcOEt): Nacalai Tesque, special grade

n-butanol (BuOH): manufactured by Nacalai Tesque, special grade Arabic gum: manufactured by Wako Pure Chemical Industries, Ltd. Carboxymethylcellulose sodium salt (CMC-Na): manufactured by Wako Pure Chemical Industries, Ltd. carbon powder (activated carbon; powder): Japanese Porcine pancreatic lipase manufactured by Kojun Pharmaceutical Co., Ltd .: Sigma-Aldrich

Silica gel for column chromatography (SiO ₂ ): Fuji Silysia Chemical, BW-200, 150 to 350 mesh Reversed phase silica gel for column chromatography: Fuji Silysia Chemical, Chromatorex ODS DM1020T, 100 to 200 mesh
ODS column for HPLC: Intertsil ODS-3 (250 × 20mm (inner diameter)) manufactured by GL Sciences Inc.

NMR: JEOL Datum (JEOL), ECA-600K (600 MHz)
IR: FTIR-8100, manufactured by Shimadzu Corporation
HPLC: Shimadzu Corporation detector, differential refractive index detector RID-6A
UV detector SPD-10A
Liquid feeding unit; LC-6AD
GC: Shimadzu GC-14A
MS: JEOL (JEOL), FABMS, HRFABMAS; JMS-SX 102A
EIMS, HREIMS; JMS-GCMATE

  In nuclear magnetic resonance (NMR) spectra, chemical shift δ is expressed in parts per million (ppm), and abbreviations have the following meanings: s: singlet; d: doublet; dd: double doublet; t : Triplet; q: quartet; dq: double quartet; Ara; α-L-arabinopyranosyl group; Glc: β-D-glucopyranosyl group, Rha; α-L-rhamnopyranosyl group.

Example 1
(1) Extraction of mate and preparation of ethyl acetate soluble fraction and n-butanol soluble fraction A dry leaf part (3.0 kg) of mate (Ilex paraguariensis) was repeatedly extracted with MeOH (30 L) three times with heat. The extract was filtered, MeOH was newly added to the residue, and the same extraction operation was performed three times. The MeOH extracts were combined and the solvent was distilled off under reduced pressure to obtain a MeOH extract (1.0 kg, yield 33.3%).
500.0 g of the obtained MeOH extract was suspended in H ₂ O ( ₂ L) and then partitioned and extracted three times with EtOAc (2 L). The H ₂ O soluble fraction was extracted three times with n-BuOH (2 L). Partition extraction was performed, and each soluble fraction was evaporated under reduced pressure to obtain an EtOAc-soluble fraction extract (168.3 g, 11.2%), an n-BuOH soluble fraction extract (251.1 g, 16.7%), H ₂ O soluble fraction extract (96.5 g, 6.4%) was obtained.

(2) Purification of AcOEt soluble fraction The obtained EtOAc soluble fraction extract (150.0 g) was subjected to normal phase silica gel column chromatography [3.0 kg, hexane: ethyl acetate = (5: 1 → 1: 1) → acetic acid. Fr. 1 (295.7 mg), Fr. 2 (15.5 g), Fr. 3 (1.5 g), fractionated with ethyl → CHCl ₃ → CHCl ₃ : MeOH = (5: 1 → 1: 1) → MeOH] , Fr. 4 (2.7 g), Fr. 5 (1.1 g), Fr. 6 (13.3 g), Fr. 7 (0.1 g), Fr. 8 (6.8 g), Fr. 9 (3.2 g), Fr 10 (16.2 g), Fr. 11 (6.7 g), Fr. 12 (3.1 g), Fr. 13 (20.3 g), and Fr. 14 (0.1 g) were obtained.

Fr. 6 (13.3 g) was fractionated by reverse phase ODS column chromatography [390.0 g, MeOH: H ₂ O = (90:10) → MeOH], and Fr. 6-1 (599.2 mg), Fr. 6-2 (10.2 g), Fr. 6-3 [= ursolic acid (900.0 mg, 0.10%)], Fr. 6-4 (312.0 mg), Fr. 6-5 (151.3 mg), Fr. 6- 6 (993.2 mg) was obtained.
Fr. 6-2 (400.0 mg) was reversed-phase HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (90:10), flow rate : 9.0 mL / min], and isolation of 3-β-hydroxyurs-11-ene-13β (28) -orido (13.1 mg, 0.039%) and ursolic acid (308.0 mg, 0.92%) did.

Fr. 13 (20.3 g) was fractionated by reverse phase ODS column chromatography [600.0 g, MeOH: H ₂ O = (90:10) → MeOH], and Fr. 13-1 (994.1 mg), Fr. 13-2 (197.7 mg), Fr. 13-3 (506.8 mg), Fr. 13-4 (10.6 g), Fr. 13-5 (509.4 mg), Fr. 13-6 (1.1 g), Fr. 13-7 (477.0 mg), Fr. 13-8 (662.8 mg), Fr. 13-9 (1.2 g), and Fr. 13-10 (3.5 g) were obtained.
Fr. 13-6 (500.0 mg) was reversed-phase HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (45:55), flow rate : 9.0 mL / min], and rutin (48.7 mg, 0.012%) and kaempferol-3-O-rutinoside (32.8 mg, 0.0079%) were isolated.

Fr. 13-8 (662.8 mg) was reversed-phase HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (35:65), flow rate : 9.0 mL / min], matenoside B (11.3 mg, 0.0013%) was isolated. Fr. 13-9 (250.0 mg) was reversed-phase HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (35:65), flow rate : 9.0 mL / min], matesaponin 1 (21.6 mg, 0.010%) was isolated.
All of the above known compounds were identified by comparing ¹ H-NMR and ¹³ C-NMR spectrum data of these compounds with literature values.

(3) Purification of n-BuOH soluble fraction The obtained n-BuOH soluble fraction extract (150.0 g) was subjected to normal phase silica gel column chromatography [3.0 kg, CHCl ₃ → CHCl ₃ : MeOH: H ₂ O = Fr. 1 (14.7 g), Fr. 2 (26.9 g), Fr. 3 Fr. 1 (14.7 g), Fr. 2 (26.9 g), Fr. 3 (30.8 g), Fr. 4 (41.3 g), Fr. 5 (34.5 g), and Fr. 6 (5.5 g) were obtained.

Fr. 1 (14.0 g) was subjected to reverse phase ODS column chromatography [420.0 g, MeOH: H ₂ O = (10:90) → (30:70) → (50:50) → (70:30) → MeOH] Fr. 1-1 (438.4 mg), Fr. 1-2 [= Theobromine (1.3 g, 0.15%)], Fr. 1-3 [= Caffeine e (3.3 g, 0.38%) ], Fr. 1-4 [= Caffeine (928.5 mg, 0.38%)], Fr. 1-5 (1.2 g), Fr. 1-6 (755.7 mg), Fr. 1-7 (1.0 g), Fr. 1-8 (282.9 mg), Fr. 1-9 (352.3 mg), Fr. 1-10 (70.3 g), Fr. 1-11 (1.5 g), Fr. 1-12 (430.0 mg) Obtained.
Caffeine (567.6 mg, 0.065%) was isolated from the Fr. 1-5 (1.2 g) by recrystallization (MeOH).

Fr. 2 (25.0 g) was subjected to reverse phase ODS column chromatography [750.0 g, MeOH: H ₂ O = (10:90) → (30:70) → (50:50) → (70:30) → (80 Fr. 2-1 (663.2 mg), Fr. 2-2 (4.4 g), Fr. 2-3 (829.0 mg), Fr. 2-4 (1.7 g ), Fr. 2-5 (1.1 g), Fr. 2-6 (748.4 mg), Fr. 2-7 (157.0 mg), Fr. 2-8 (446.2 mg), Fr. 2-9 (442.0 mg) ), Fr. 2-10 (260.2 mg), Fr. 2-11 (9.0 g), Fr. 2-12 (1.7 g), Fr. 2-13 (1.5 g), 2-14 (237.5 mg) Obtained.

Fr. 2-7 (157.0 mg) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (60:40), flow rate: 9.0 (R) -linalyl 6-O-arabinopyranosyl glucopyranoside (74.8 mg, 0.090%) was isolated. Fr. 2-11 (500.0 mg) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (70:30), flow rate: 9.0 mL / min] was used for isolation and purification to isolate matesaponin 1 (31.4 mg, 0.051%).

Fr. 2-14 (237.5 mg) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inside diameter)), eluent: CH ₃ CN: MeOH: [H ₂ O: EtOAc = (99 : 1)] = (35:16:49), flow rate: 9.0 mL / min], matesaponin 1 (27.7 mg, 0.029%), matesaponin J3b (17.4 mg, 0.060%), matesaponin J3a ( 30.3 mg, 0.10%), 3-O-β-D-glucopyranosyl (1 → 3) -α-L-2-O-acetylarabinopyranosyl ursolic acid 28-O-β-D-glucopyranoside (13.6 mg , 0.047%) was isolated.

Fr. 3 (30.0 g) was subjected to reverse phase ODS column chromatography [900.0 g, MeOH: H ₂ O = (10:90) → (30:70) → (50:50) → (70:30) → (80 Fr. 3-1 (404.4 mg), Fr. 3-2 (1.8 g), Fr. 3-3 (1.7 g), Fr. 3-4 (426.1 mg ), Fr. 3-5 [= rutin (2.2 g, 0.25%)], Fr. 3-6 [= kaempferol-3-O-rutinoside (1.5 g, 0.17%)], Fr. 3-7 (2.4 g), Fr. 3-8 (1.2 g), Fr. 3-9 (5.1 g), Fr. 3-10 (11.0 g), Fr. 3-11 (1.7 g), Fr. 3-12 (465.0 mg).

Fr. 3-8 (1.2 g) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inside diameter)), eluent: MeOH: H ₂ O = (65:35), flow rate: 9.0 The product was separated and purified using [mL / min] to isolate matenoside A (177.2 mg, 0.020%). Fr. 3-9 (1.0 g) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inside diameter)), eluent: MeOH: H ₂ O = (70:30), flow rate: 9.0 mL / min] was used for separation and purification, and 3-O-α-L-rhamnopyranosyl (1 → 2) -α-L-arabinopyranosyl oleanolic acid 28-O-β-D-glucopyranosyl (1 → 6 ) -β-D-glucopyranoside (363.1 mg, 0.21%) was isolated.

Fr. 3-10 (2.0 g) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (70:30), flow rate: 9.0 mL / min], and purified Fr. 3-10-1 (198.6 mg), Fr. 3-10-2 (237.1 mg), Fr. 3-10-3 [= nudicaucin C (215.9 mg, 0.10%)], Fr. 3-10-4 (560.4 mg) was obtained.

Fr. 3-10-1 (198.6 mg) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (70:30), flow rate : 9.0 mL / min], and 3-O-α-L-rhamnopyranosyl (1 → 2) -α-L-arabinopyranosyl oleanolic acid 28-O-β-D-glucopyranosyl (1 6) -β-D-glucopyranoside (67.3 mg, 0.007%) was isolated.

Fr. 3-10-2 (237.1 mg) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (70:30), flow rate : 9.0 mL / min] and purified Fr. 3-10-2-1 (53.5 mg) and Fr. 3-10-2-2 [= nudicaucin C (36.4 mg, 0.017%)] Released. Fr. 3-10-2-1 (53.5 mg) by HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (69:31) , Flow rate: 9.0 mL / min], and matesaponin 2 (6.3 mg, 0.0029%) was isolated.

Fr. 4 (30.0 g) was subjected to reverse phase ODS column chromatography [1.2 kg, MeOH: H ₂ O = (10:90) → (30:70) → (50:50) → (70:30) → (80 Fr. 4-1 (2.5 g), Fr. 4-2 (4.3 g), Fr. 4-3 (13.2 g), Fr. 4-4 (5.2 g ), Fr. 4-5 [= rutin (2.2 g, 0.25%)], Fr. 4-6 (1.2 g), Fr. 4-7 (956.8 mg), Fr. 4-8 (2.7 g), Fr 4-9 (2.9 g), Fr. 4-10 (1.7 g), Fr. 4-11 (786.2 mg), Fr. 4-12 (601.4 mg), Fr. 4-13 (1.6 g) were obtained. It was.

Fr. 4-3 (4.0 g) was subjected to reverse phase ODS column chromatography [120.0 g, MeOH: H ₂ O = (10:90) → (20:80) → (30:70) → (40:60) → (50:50) → MeOH], and 3,5-O-dicaffeoylquinic acid (2.6 g, 0.32%) was isolated.
Fr. 4-7 (500.0 mg) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inside diameter)), eluent: MeOH: H ₂ O = (60:40), flow rate: 9.0 mL / min] was used to isolate and purify matesaponin 3 (44.8 mg, 0.0099%).

Fr. 4-8 (2.4 g) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (70:30), flow rate: 9.0 mL / min], and then purified using Fr. 4-8-1 (53.8 mg), Fr. 4-8-2 (489.8 mg), Fr. 4-8-3 (1.2 g), Fr. 4 -8-4 (56.5 mg), Fr. 4-8-5 (82.2 mg), and Fr. 4-8-6 (392.5 mg) were obtained.

Fr. 4-8-2 (489.8 mg) by HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: CH ₃ CN: H ₂ O = (30:70) , Flow rate: 9.0 mL / min], and matesaponin 5 (72.4 mg, 0.0094%) was isolated. Fr. 4-8-3 (500.0 mg) by HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (65:35), flow rate : 9.0 mL / min], matenoside C (55.6 mg, 0.074%) was isolated.

Fr. 4-9 (1.5 g) with HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: CH ₃ CN: H ₂ O = (30:70), flow rate : 9.0 mL / min], matesaponin 5 (97.0 mg, 0.021%) and matenoside C (457.1 mg, 0.10%) were isolated.
Fr. 4-11 (786.2 mg) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (30:70), flow rate: 9.0 mL / min] was used for separation and purification. Fr. 4-11-1 (115.4 mg), Fr. 4-11-2 (58.7 mg), Fr. 4-11-3 (262.9 mg), Fr. 4 -11-4 (53.7 mg) and Fr. 4-11-5 (197.6 mg) were obtained.

Fr. 4-11-1 (115.4 mg) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (68:32), flow rate : 9.0 mL / min], and matenoside C (20.8 mg, 0.024%) was isolated.
Fr. 4-11-2 (58.7 mg) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (70:30), flow rate : 9.0 mL / min], matesaponin 4 (29.7 mg, 0.034%) was isolated.
Fr. 4-11-3 (262.9 mg) was HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (65:35), flow rate : 9.0 mL / min], and 3-O-α-L-rhamnopyranosyl (1 → 2) -α-L-arabinopyranosyl oleanolic acid 28-O-β-D-glucopyranosyl (1 → 6) -β-D-glucopyranoside (65.7 mg, 0.07%) was isolated.

Fr. 5 (33.0 g) was subjected to reverse phase ODS column chromatography [1.2 kg, MeOH: H ₂ O = (10:90) → (30:70) → (50:50) → (70:30) → MeOH] Fr. 5-1 (6.8 g), Fr. 5-2 (9.4 g), Fr. 5-3 [= 5-O-caffeoylquinic acid (3.5 g, 0.40%)], Fr. 5-4 (6.7 g), Fr. 5-5 (3.1 g), Fr. 5-6 (4.4 g), Fr. 5-7 (287.9 mg), Fr. 5-8 (1.2 g) Obtained.
Fr. 5-4 (500.0 mg) by HPLC [detection: RI, column: Inertsil ODS-3 (250 × 20 mm (inner diameter)), eluent: MeOH: H ₂ O = (10:90), flow rate: 9.0 mL / min], and purified by using 5-O-caffeoylquinic acid (20.8 mg, 0.032%), 3-O-caffeoylquinic acid (105.9 mg, 0.16%), 4-O-caffeoylquinine The acid (94.2 mg, 0.15%) was isolated.

All of the above known compounds were identified by comparing ¹ H-NMR and ¹³ C-NMR spectrum data of these compounds with literature values.
The physical property values of matenoside B found from the ethyl acetate soluble fraction and matenoside A and C found from the n-butanol soluble fraction according to the present invention as described above are shown below.

(4) Various physical properties of matenosides A to C matenoside A: yellow powder
Optical rotation: [α] _D ²⁵ + 18.7 ° (c = 0.13, MeOH)
High resolution cation FAB-MS
Theoretical value: C ₄₇ H ₇₆ O ₁₈ Na (M + Na) ⁺ : 951.4933
Actual value: 951.4938
IR (KBr, cm ^-1 ): 3600, 1716, 1650, 1070
Cation FAB-MS: m / z 951 (M + Na) ⁺
Anion FAB-MS: m / z 927 (MH) ^-

Matenoside B: White powder
Optical rotation: [α] _D ²⁹ + 47.3 ° (c = 0.63, MeOH)
High resolution cation FAB-MS
Theoretical value: C ₄₁ H ₆₆ O ₁₃ Na (M + Na) ⁺ : 789.4412
Actual value: 789.4407
IR (KBr, cm ^-1 ): 3507, 1653, 1072
Cation FAB-MS: m / z 789 (M + Na) ⁺

Matenoside C: White powder
Optical rotation: [α] _D ²⁵ −5.56 ° (c = 0.64, MeOH)
High resolution cation FAB-MS
Theoretical value: C ₅₉ H ₉₆ O ₂₆ Na (M + Na) ⁺ : 1243.6089
Actual value: 1243.6094
IR (KBr, cm ^-1 ): 3432, 1080, 1036
Cation FAB-MS: m / z 1243 (M + Na) ⁺
Anion FAB-MS: m / z 1219 (MH) ^-

Known compounds were identified by comparison of literature values of ¹ H-NMR and ¹³ C-NMR spectral data.

(5) Identification of sugar by HPLC Matenoside A (17.0 mg) was dissolved in 5% H ₂ SO ₄ aqueous solution-1,4-dioxane (1: 1, v / v, 1 ml) and heated under reflux for 2 hours. The reaction solution was neutralized with anion exchange resin Amberlite IRA-400 (trade name) (OH ^- type), the resin was filtered off, and the solvent was distilled off under reduced pressure to obtain a crude product.
The obtained product was partitioned between EtOAc and H ₂ O, and the aqueous phase fraction was subjected to HPLC analysis with an optical rotation detector under the conditions shown below, and the sugar obtained by hydrolysis was compared with the standard product. Identified. As a result, it was found that the constituent sugars of matenoside A were L-arabinose and D-glucose.
The same treatment was performed for matenoside B (3.5 mg) and matenoside C (3.3 mg). As a result, it was found that the constituent sugars of matenoside B were L-arabinose and D-glucose.
It was also found that the constituent sugars of matenoside C were L-arabinose, D-glucose and L-rhamnose.

HPLC conditions
Detector: Shodex OR-2
Column: Kaseisorb LC NH2-60-5 (4.6 x 250 mm (inner diameter), 5μm)
Solvent: MeCN-H ₂ O (80:20, v / v)
Flow rate: 0.8 ml / min
Injection volume: 20 μL
Column temperature: room temperature

Holding time
L-arabinose: 8.3 minutes
D-glucose: 9.5 minutes
L-rhamnose: 6.5 minutes

Matenoside A, B and C ¹ H-NMR (600 MHz) data and ¹³ C-NMR (150 MHz) data are summarized in the following table.

Regarding the novel saponin compound, matenoside A, matenoside B, and matenoside C found by the present invention, a structural formula in which chemical shift values of ¹ H-NMR and ¹³ C-NMR are entered in the respective chemical structural formulas; The structural formula showing the H correlation is shown below together with the respective physical data.

Test example 1
Method for evaluating lipase inhibitory activity of matecha extract Triolein solution (triolein 80 mg, phosphatidylcholine 10 mg, taurocholic acid 5 mg as a substrate, add 9 mL of 0.1 M Tris buffer (containing 0.1 M NaCl, pH 7.0), 10 The solution was sonicated for 5 minutes] 5 μL of the test sample solution was added to 100 μL, and 95 μL of Tris buffer (pH 7.0) was added to make a total volume of 200 μL. After preheating at 37 ° C. for 3 minutes, 50 μL of lipase solution (derived from porcine pancreas) was added and allowed to react for 30 minutes, and then placed in a boiling water bath (90 to 100 ° C.) for 2 minutes to stop the reaction.
Separately, 50 μL of Tris buffer was added to each sample in place of the lipase solution, and the same operation was performed as a blind preparation.
The amount of oleic acid produced was measured by the NEFA C kit method (NEFA C-Test Wako: manufactured by Wako Pure Chemical Industries).
The lipase was dissolved in Tris buffer and adjusted to a concentration that produced about 0.7 meq / L oleic acid under the experimental conditions. The test sample was dissolved and diluted with DMSO. Orlistat (Xenical: Trademark) was used as a comparative control drug.

  In the following, matecha methanol extract, ethyl acetate soluble fraction, n-butanol soluble fraction, matenoside C, matenosaponin 1, nudicaucin C and 3-O-α-L-rhamnopyranosyl (1 → 2) -α- The results of lipase inhibitory activity at various concentrations of L-arabinopyranosyl oleanolic acid 28-O-β-D-glucopyranosyl (1 → 6) -β-D-glucopyranoside are shown.

  From the above results, in the lipase inhibitory activity test, matecha methanol extract, ethyl acetate soluble fraction, n-butanol soluble fraction, matenoside C, matenosaponin 1, nudicaucin C and 3-O-α-L-rhamnopyranosyl (1 → 2) -α-L-arabinopyranosyl oleanolic acid 28-O-β-D-glucopyranosyl (1 → 6) -β-D-glucopyranoside has lipase inhibitory activity and is a free fatty acid It was found to have a production-inhibiting effect.

Example 2
In accordance with a method known in the art, 10 parts by weight of extract extract was mixed with 25 parts by weight of lactose and filled into a gelatin capsule to obtain a gelatin capsule containing 500 mg of the extract in one capsule.

Example 3
A gelatin capsule was obtained in the same manner as in Example 2, except that the ethyl acetate-soluble fraction obtained in Example 1 was replaced with the extract of Example 2.

Example 4
A gelatin capsule was obtained in the same manner as in Example 2, except that the concentrate of the n-BuOH soluble fraction obtained in Example 1 was replaced with the extract of Example 2.

Example 5
50 parts by weight of rice flour, 5 parts by weight of sugar, 10 parts by weight of whole eggs, and 1 part by weight of the extract of the material obtained in Example 1 were weighed.
After mixing sugar with whole eggs, rice flour previously passed through a sieve was added and lightly mixed to make a dough, which was shaped into an appropriate shape and baked in an oven to make a rice cracker.

Example 6
A rice cracker was prepared in the same manner as in Example 5, except that the ethyl acetate soluble fraction obtained in Example 1 was replaced with the extract of Example 5.

Example 7
A rice cracker was produced in the same manner as in Example 5 except that the concentrate of the n-BuOH soluble fraction obtained in Example 1 was replaced with the extract of the texture of Example 5.

Example 8
100 parts by weight of flour, 4 parts by weight of salt, 5 parts by weight of the extract of the texture obtained in Example 1 and 45 parts by weight of water were weighed and mixed well according to a conventional method to produce udon.

Example 9
Udon was produced in the same manner as in Example 8, except that the ethyl acetate-soluble fraction obtained in Example 1 was replaced with the extract of Example 8.

Example 10
Udon was produced in the same manner as in Example 8, except that the concentrate of the n-BuOH soluble fraction obtained in Example 1 was replaced with the extract of the texture of Example 8.

Example 11
According to the following prescription, 100 mL of soft drink (drink) was prepared.
Matecha extract 1 (weight / volume)%
Dietary fiber 5 (weight / volume)%
Vitamin B ₂ 0.002 (weight / volume)%
Vitamin B ₆ 0.005 (weight / volume)%
Vitamin C 0.1% (weight / volume)
Fructose dextrose liquid sugar 5 (weight / volume)%
Appropriate amount of water

Example 12
A drink was produced in the same manner as in Example 11, except that the ethyl acetate-soluble fraction obtained in Example 1 was replaced with the extract of Example 11.

Example 13
A drink was produced in the same manner as in Example 11, except that the n-butanol soluble fraction obtained in Example 1 was replaced with the extract of Example 11.

Example 14
According to the following prescription, 100 g of health food (capsule) was prepared.
Matecha extract 30% by weight
GABA 30% by weight
Theanine 30% by weight
Vitamin C 30% by weight
Lactose 30% by weight

Example 15
Capsules were produced in the same manner as in Example 14, except that the ethyl acetate-soluble fraction obtained in Example 1 was replaced with the material extract of Example 14.

Example 16
Capsules were produced in the same manner as in Example 14, except that the n-butanol-soluble fraction obtained in Example 1 was replaced with the extract of Example 14.

  A composition comprising at least one of the matecha extract according to the present invention or the saponin compound according to the present invention contained in the extract has a lipase inhibitory activity, and is a free fatty acid production inhibitor, anti-obesity It can be safely used as a composition for antilipidemia and antimetabolic syndrome.

Claims (10)

The following general formula (1):

[Where:
R ¹ is an α-L-arabinopyranosyl group, β-D-glucopyranosyl- (1 → 3) -α-L-arabinopyranosyl group or α-L-rhamnopyranosyl- (1 → 2)- O- [β-D-glucopyranosyl- (1 → 3)]-α-L-arabinopyranosyl group,
R ² is a β-D-glucopyranosyl group or β-D-glucopyranosyl- (1 → 6) -β-D-glucopyranosyl group,
R ³ and R ⁴ are each independently a hydrogen atom or a methyl group,
R ⁵ is a methyl group or a hydroxymethyl group]
A saponin compound represented by: In the compound of the formula (1), R ¹ is β-D-glucopyranosyl- (1 → 3) -β-D-arabinopyranosyl group, R ² is β-D-glucopyranosyl group, R The saponin compound according to claim 1, wherein ³ is a methyl group, R ⁴ is a hydrogen atom, and R ⁵ is a hydroxymethyl group. In the compound of the formula (1), R ¹ is an α-L-arabinopyranosyl group, R ² is a β-D-glucopyranosyl group, R ³ is a methyl group, and R ⁴ is a hydrogen atom. The saponin compound according to claim 1, wherein R ⁵ is a hydroxymethyl group. In the compound of the above formula (1), R ¹ is α-L-rhamnopyranosyl- (1 → 2) -O- [β-D-glucopyranosyl- (1 → 3)]-α-L-arabinopyranosyl group R ² is a β-D-glucopyranosyl- (1 → 6) -β-D-glucopyranosyl group, R ³ is a hydrogen atom, R ⁴ is a methyl group, and R ⁵ is a methyl group. The saponin compound according to claim 1.   Extract the material with water or water-containing lower alcohol to obtain an extract, and if necessary, contain an organic solvent soluble fraction obtained by partitioning this extract with ethyl acetate / water and n-butanol / water. A mate extract comprising at least one compound according to claims 1-4.   The extract according to claim 5, wherein the material is a dried leaf portion of a material that is a genus Ilexaceae.   A pharmaceutical composition comprising at least one of the compounds according to claims 1 to 4 or the extract according to claim 5 or 6 as an active ingredient.   The composition of Claim 7 used for free fatty acid production suppression.   The composition according to claim 7, which is used for anti-obesity, anti-hyperlipidemia or fatty liver improvement.   A health food comprising at least one of the compounds according to claims 1 to 4, the extract according to claim 5 or 6, or the composition according to claim 7.