JP2005132810A - Physiologically active substance from nameko and/or maitake mushrooms and producing method and agency - Google Patents
Physiologically active substance from nameko and/or maitake mushrooms and producing method and agency Download PDFInfo
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Abstract
Description
本発明は、ナメコ、および/または、マイタケの食用茸から得られた、抗癌活性、抗アレルギー、免疫賦活性、抗酸化活性などの生理活性を持つ物質およびその製造方法並びに当該生理活性物質を含有する製剤に関するものである. The present invention relates to a substance having a physiological activity such as anticancer activity, antiallergic activity, immunostimulatory activity, antioxidant activity, etc., obtained from an edible candy of sea cucumber and / or maitake, a method for producing the same, and the physiologically active substance. It relates to the preparations contained.
本発明者らは、永年にわたり茸類の生理活性について詳しく研究を行っており、その結果、ある種の茸類に、抗癌作用、免疫賦活作用、抗酸化作用、血圧及び血糖低下作用などの生理活性を示すことがしられるようになっている.しかし、茸の種類によっては、それらの活性がないか、非常に弱いにも係らず、よく効くと過度に効果が宣伝され、発売されている. The present inventors have been studying in detail the physiological activity of mosses for many years. As a result, anti-cancer action, immunostimulatory action, antioxidant action, blood pressure, blood glucose lowering action, etc. It is designed to show physiological activity. However, depending on the type of cocoon, although it is not active or very weak, if it works well, the effect is over-advertised and released.
本発明者らは、膨大な研究データに基づく確実な研究結果から、特に茸の抗癌作用が、含まれる多糖体によるものであることを知り、茸の多糖体であるグルカンについて研究した.しかしながら、純粋な単純多糖体、グルカンは、注射によっては活性を示すが、経口では効果を示さないことが判明した。
注射による投与は、病院でしか行い得ず、また、経口での投与の方が投与を受けるものにとって苦痛が少ないため、経口で有効な茸の活性成分の提供が強く求められている.The present inventors have studied the glucan which is a polysaccharide of sputum, knowing that the anticancer action of sputum is due to the contained polysaccharide, especially from the reliable research results based on a huge amount of research data. However, the pure simple polysaccharide, glucan, was found to be active by injection but not to be effective orally.
Administration by injection can only be performed in hospitals, and since oral administration is less painful for those receiving the administration, there is a strong need to provide an active ingredient for orally effective sputum.
本発明の課題は、経口投与によって抗癌作用、免疫賦活作用などの生理活性を有する物質を高濃度に含有する茸類を選択し、それらの生理活性物質を効率よく採取する方法を提供するとともに、それら得られた活性物質を使用に適するようにした製剤を提供するものである. An object of the present invention is to provide a method for selecting moss containing a high concentration of substances having physiological activities such as anticancer action and immunostimulatory action by oral administration, and efficiently collecting those physiologically active substances. The present invention provides a preparation in which the obtained active substances are suitable for use.
本発明者らは、上記の課題を解決するため種々検討した結果、食用に供されている茸類、ナメコ、および/または、マイタケの子実体および/または菌糸体が、経口で有効な活性物質を多量に含有し、それらの茸類の水もしくは炭素数1から4の低級アルコールのような親水性溶媒またはこれらの混合溶媒での抽出物には経口で有効な活性物質が含有されており、これらの抽出物をさらに分子フルイ法により処理し、低分子部分及び高分子部分を除いた、一定分子量範囲のものは、優れた生理活性を示すことを見出し、本発明を完成した. As a result of various studies to solve the above-mentioned problems, the present inventors have found that fruit bodies and / or mycelia of maitake, which are used for food, are orally active active substances. And the extract of the moss with a hydrophilic solvent such as water or a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof contains an orally active active substance, These extracts were further processed by the molecular sieve method, and it was found that those having a certain molecular weight range excluding the low molecular part and the high molecular part showed excellent physiological activity, and the present invention was completed.
すなわち本発明は、ナメコ、および/または、マイタケの茸を水、親水性溶媒もしくはこれらの混合溶媒で抽出し、次いで、これらの抽出物を分子フルイ法により処理し、分子量6,000〜100,000の部分を採取することにより得られる生理活性物質EEM−Pを提供するものである. That is, the present invention is to extract nameko and / or maitake mushrooms with water, a hydrophilic solvent or a mixed solvent thereof, and then treat these extracts by a molecular sieve method to obtain a molecular weight of 6,000 to 100, A bioactive substance EEM-P obtained by collecting 000 parts is provided.
また本発明の、生理活性物質EEM−Pはナメコ、および/または、マイタケのような食用に供される茸類の子実体および/または菌糸体の少なくとも一つの微粉砕物(以下、「茸」と記載する)を水および/または親水性溶媒を用いて抽出することにより得られる.この抽出に用いられる親水性溶媒としては、炭素数1から4の低級アルコールが好ましく使用される. In addition, the bioactive substance EEM-P of the present invention is a pulverized fruit body and / or mycelium of moss used for edible use such as nameko and / or maitake (hereinafter referred to as “茸”). Is extracted with water and / or a hydrophilic solvent. As the hydrophilic solvent used for this extraction, a lower alcohol having 1 to 4 carbon atoms is preferably used.
上記茸から生理活性物質EEM−Pを含む画分の抽出は、例えば、原料である茸に適量の水またはアルコール類の親水性溶媒を添加し、例えば、80ないし98℃程度、好ましくは、水の場合は90から98℃の温度条件で、還流下に、1ないし4時間程度、好ましくは、2〜4時間抽出することにより行われる.抽出にあたっては、茸の重量に対して、3ないし10重量倍程度の溶媒が使用される. Extraction of the fraction containing the physiologically active substance EEM-P from the cocoon, for example, by adding an appropriate amount of water or a hydrophilic solvent of alcohols to the cocoon as a raw material, for example, about 80 to 98 ° C., preferably water In this case, the extraction is performed at 90 to 98 ° C. under extraction under reflux for about 1 to 4 hours, preferably 2 to 4 hours. In the extraction, about 3 to 10 times by weight of solvent is used with respect to the weight of the soot.
上記のようにして得られた抽出液は、必要によりろ過等で固形不純物を分離した後、減圧下に溶媒を溜去することにより、固形物として抽出物を採取することができる. The extract obtained as described above can collect the extract as a solid by separating solid impurities by filtration or the like, if necessary, and then distilling off the solvent under reduced pressure.
このようにして得られた固形物は、それ自体でも経口投与で活性を示し、そのままでも使用できるが、活性を高めるため、抽出液を分子フルイ処理して不要の部分を除去し、再度減圧下に濃縮して得られた分子量6,000〜100,000の部分を、生理活性物質EEM−Pとして使用する. The solid obtained in this way is active by oral administration and can be used as it is. However, in order to increase the activity, the extract is subjected to molecular sieve treatment to remove unnecessary parts, and again under reduced pressure. A portion having a molecular weight of 6,000 to 100,000 obtained by concentrating to 1 is used as the bioactive substance EEM-P.
上記の目的のために使用される分子フルイとしては、分子フルイ膜(平膜)、中空ろ過膜、透析膜、分子フルイクロマト等を挙げることができ、生理活性物質EEM−Pの分取取得は、これらを用いる方法を適宜応用することができる. Examples of molecular sieves used for the above purpose include molecular sieve membranes (flat membranes), hollow filtration membranes, dialysis membranes, molecular sieve chromatography, etc. The methods using these can be applied as appropriate.
以下、モジュールタイプの中空ろ過を用いた場合を例にとって生理活性物質EEM−Pの分取取得方法を説明すれば次の通りである.すなわち、前記のように抽出して得た茸の水および/または親水性溶媒抽出液(生理活性物質EEM−Pを含む画分)を、まず、低分子分画モジュール、例えば、AIP−3013、AIP−2013,またはAIP−1010(旭化成工業社製)で処理して低分子物質(分子量6,000以下の物質)を除く、次いで、低分子物質が除去された濃縮液を高分子分画モジュール、例えば、AIP−3013、AIP−2013,またはAIP−1010(旭化成製)で処理して高分子物質(分子量100,000以上の物質)を除き、外液を再び低分子分画モジュールを用いて濃縮する.かくするころにより、分子量が6,000〜100,000の生理活性物質EEM−Pを得ることができる. Hereinafter, taking a case where module type hollow filtration is used as an example, a fractionation acquisition method of the physiologically active substance EEM-P will be described as follows. That is, the water and / or hydrophilic solvent extract obtained by extraction as described above (a fraction containing the bioactive substance EEM-P) is first subjected to a low molecular fraction module such as AIP-3013, A low molecular weight material (a material having a molecular weight of 6,000 or less) is removed by treatment with AIP-2013 or AIP-1010 (manufactured by Asahi Kasei Kogyo Co., Ltd.). For example, by treating with AIP-3013, AIP-2013, or AIP-1010 (manufactured by Asahi Kasei) to remove the high molecular substance (substance having a molecular weight of 100,000 or more), the external liquid is again used with the low molecular fraction module. Concentrate. By doing so, a physiologically active substance EEM-P having a molecular weight of 6,000 to 100,000 can be obtained.
なお、一般に分子フルイ法によるときは、分子の形状によってフルイにかかり方が異なり、他の方法で求めた分子量と異なる場合がある.従って、本発明における、分子量とは、分子フルイ法によるものを意味し、より正確にいうなら、低分子量の分子量は、AIP−3013ないしはこれと同等の低分子分画モジュールを使用して測定した値である。同様、高分子分側の分子量は、AIP−3013ないしはこれと同等の低分子分画モジュールを使用して測定した値である. In general, the molecular sieve method differs depending on the shape of the molecule and may differ from the molecular weight obtained by other methods. Accordingly, the molecular weight in the present invention means the molecular sieve method, and more precisely, the low molecular weight molecular weight was measured using AIP-3013 or a low molecular fraction module equivalent thereto. Value. Similarly, the molecular weight on the high molecular weight side is a value measured using AIP-3013 or a low molecular weight fraction module equivalent thereto.
このようにして得られた茸類の生理活性物質EEM−Pと記したが、茸からの生理活性物質は、茸菌糸体を原料として使用する場合、培養条件によってその収率が異なり、培地、培養温度などを最適に選ぶ必要がある.また、茸の種類によっても若干異なる物理化学的性状を示すことがある. Although the bioactive substance EEM-P of moss obtained in this manner was described, the yield of the bioactive substance from cocoons varies depending on the culture conditions when the koji mycelium is used as a raw material, It is necessary to select the optimal culture temperature. In addition, physicochemical properties may be slightly different depending on the type of cocoon.
例えば、ナメコを原料として製造した生理活性物質EEM−P(以下、EEM−PNと記す)は、後記実施例1に示す物理化学的性状を示す.また、マイタケを原料として製造した生理活性物質EEM−P(以下、EEM−PMと記す)は、後記実施例2に示す物理化学的性状を示すものである. For example, a bioactive substance EEM-P (hereinafter referred to as EEM-PN) produced from nameko as a raw material exhibits the physicochemical properties shown in Example 1 below. Moreover, the bioactive substance EEM-P (hereinafter referred to as EEM-PM) produced using maitake as a raw material exhibits the physicochemical properties shown in Example 2 below.
このように、物理化学的性質においては、若干の相違があっても、いずれも後記実施例3に示すように、各茸の単純な熱水抽出物と比べより高い生理活性を有するものであって、共通な性質を保有するものといえる. Thus, even if there is a slight difference in physicochemical properties, as shown in Example 3 below, each has a higher physiological activity than the simple hot water extract of each cocoon. Therefore, it can be said that they possess common properties.
以上説明した生理活性物質EEM−Pは、経口投与により生理活性を発揮しうるものが、所期の効果を発揮させるための経口摂取量は、人の年齢、体重、目的などにより異なるが、一般的に、成人1日あたりEEM−Pとして200〜5000mgの範囲、好ましくは、1000〜3000mgの範囲が好ましく、この量を1日数回に分けて服用することが適当であろう. Although the biologically active substance EEM-P described above can exert physiological activity by oral administration, the oral intake for exerting the desired effect varies depending on the age, weight, purpose, etc. of the person, In particular, EEM-P per day for an adult is in the range of 200 to 5000 mg, preferably 1000 to 3000 mg, and it is appropriate to take this amount divided into several times a day.
なお、本発明の生理活性物質EEM−Pは、常法により錠剤、顆粒剤、カプセル剤、液剤として使用できるが、そのまま大気中に放置すると吸湿して変色、変質することがあるので、フィルムコートを施した錠剤として使用することが好ましい.この目的のために使用されるフィルムコート材としては、大豆ペプチド、貝殻樹脂材等が好ましく、他のフィルムコート材では十分に吸湿を防止し得ない場合があり、変色、変質を生じることもあり得る。 The physiologically active substance EEM-P of the present invention can be used as tablets, granules, capsules, and liquids by a conventional method. However, if it is left in the air as it is, it may absorb moisture and change color or change its quality. It is preferable to use as tablets with The film coating material used for this purpose is preferably soy peptide, shell resin material, etc., and other film coating materials may not be able to sufficiently prevent moisture absorption and may cause discoloration and alteration. obtain.
以下に実施例を示して本発明を更に具体的に説明する.なお実施例においては、生理活性として抗癌効果を示したが、本願物質は、抗癌性に限定されるものでなく、前記の生理活性を有するものであることはいうまでもない. The present invention will be described more specifically with reference to the following examples. In the examples, the anticancer effect was shown as the physiological activity. However, it goes without saying that the substance of the present application is not limited to the anticancer property and has the aforementioned physiological activity.
実施例 1
ナメコ1000グラムを細かく粉砕した後、熱水抽出し、得られた抽出液から沈殿物をPS−88膜(大塚実業社製)で除去する.この熱水抽出液を低分子分画用モジュールタイプの中空ろ過膜、AIP−3013(旭化成工業社製)を用いて高分子物質(分子量100,000以上の物質)を含有する画分を濃縮した.その濃縮液を高分子分画用モジュールタイプの中空ろ過膜、AIP−3013(旭化成工業社製)を用いて高分子物質(分子量100,000以上)除き、その外液を再び低分子分画膜モジュールAIP−3013を用いて、濃縮した.このように連続的処理して、分子量4,000〜100,000め生理活性物質EEM−PNを得た.
[EEM−PNの物理化学的性質]
(1)紫外線吸収(水溶液中370〜190nmのUV吸収を測定):
極大値:258.0nm
(2)赤外線吸収(ATR法で測定):
極大値(cm−1):3235,1556,1396,1033
(吸収曲線を図1として示す)
(3)1H−NMR(D2O中で測定):
チャートを図3として示す.
(4)13C−NMRD2O中で測定):
チャートを図5に示す.
(5)蛋白含量
42.9%
(6)糖含量
19.1%
(7)主たる糖の構成比
グルコース:ガラクトース:マンノース=3:1:1
(8)アミノ酸組成
アスパラギン酸:12.2%、グルタミン酸:13.9%、グリシン:10.9%
、アラニン:11.3%、バリン:6.8%、セリン:6.5%、ロイシン:6.
9%。
こうして得られたEEM−PNは、凍結乾燥することにより固形粉末とすることができ、更にそれを打錠することにより、経口投与可能な錠剤となしえた.Example 1
After pulverizing 1000 grams of nameko finely, hot water extraction is performed, and a precipitate is removed from the obtained extract with a PS-88 membrane (manufactured by Otsuka Business Co., Ltd.). Using this hot water extract, a fraction containing a high molecular weight substance (a substance having a molecular weight of 100,000 or more) was concentrated using a low molecular fraction module-type hollow filtration membrane, AIP-3013 (manufactured by Asahi Kasei Corporation). . The concentrated solution was removed from the polymer material (molecular weight of 100,000 or more) using a module-type hollow filtration membrane for polymer fractionation, AIP-3013 (manufactured by Asahi Kasei Kogyo Co., Ltd.), and the external solution was again removed from the polymer solution with a low molecular fraction. Concentrated using module AIP-3013. In this way, a bioactive substance EEM-PN having a molecular weight of 4,000 to 100,000 was obtained.
[Physicochemical properties of EEM-PN]
(1) UV absorption (measurement of UV absorption at 370 to 190 nm in an aqueous solution):
Maximum value: 258.0 nm
(2) Infrared absorption (measured by ATR method):
Maximum value (cm −1 ): 3235, 1556, 1396, 1033
(The absorption curve is shown as FIG. 1)
(3) 1 H-NMR (measured in D 2 O):
The chart is shown in Fig. 3.
(4) Measured in 13 C-NMRD 2 O):
Figure 5 shows the chart.
(5) Protein content 42.9%
(6) Sugar content 19.1%
(7) Composition ratio of main sugars Glucose: galactose: mannose = 3: 1: 1
(8) Amino acid composition Aspartic acid: 12.2%, Glutamic acid: 13.9%, Glycine: 10.9%
, Alanine: 11.3%, valine: 6.8%, serine: 6.5%, leucine: 6.
9%.
The thus obtained EEM-PN could be made into a solid powder by lyophilization, and further tableted to form an orally administrable tablet.
実施例 2
マイタケ1000グラムを熱水で還流しながら抽出し、熱水抽出物107グラムを得る.これを水に溶解した後、透析チューブに入れて透析し、透析内膜の高分子物質(分子量60,000以上)を含む画分を得た.この高分子物質を含む画分を分子フルイ膜PM−10で処理して高分子物質(分子量50,000以上)を除き、凍結乾燥して、分子量が6,000〜50,000の生理活性物質EEM−PM高含量物質を得た.これを粉末剤とした.
[EEM−PMの物理化学的性質]
(1)紫外線吸収(水溶液中370〜190nmのUV吸収を測定):
極大値:258.4nm
(2)赤外線吸収(ATR法で測定):
極大値(cm−1):3220,1572,1395,987
(吸収曲線を図2に示す)
(3)1H−NMR(D2O中で測定):
チャートを図4に示す.
(4)蛋白含量
64.8%
(5)糖含量
15.7%
(6)主たる糖の構成比
グルコース:ガラクトース:マンノース=1:2:1
(7)アミノ酸組成
アスパラギン酸:12.6%、グルタミン酸:13.2%、グリシン11.2%、アラニン:10.1%、バリン:6.7%、セリン:6.6%、ロイシン:6.7%。Example 2
Extract 1,000 grams of maitake with refluxing with hot water to obtain 107 grams of hot water extract. This was dissolved in water, then put into a dialysis tube and dialyzed to obtain a fraction containing a polymer substance (molecular weight of 60,000 or more) in the dialysis inner membrane. The fraction containing the polymer substance is treated with a molecular sieve membrane PM-10 to remove the polymer substance (molecular weight of 50,000 or more), freeze-dried, and a physiologically active substance having a molecular weight of 6,000 to 50,000. A material with a high EEM-PM content was obtained. This was used as a powder.
[Physicochemical properties of EEM-PM]
(1) UV absorption (measurement of UV absorption at 370 to 190 nm in an aqueous solution):
Maximum value: 258.4 nm
(2) Infrared absorption (measured by ATR method):
Maximum value (cm −1 ): 3220, 1572, 1395, 987
(The absorption curve is shown in FIG. 2)
(3) 1 H-NMR (measured in D 2 O):
Figure 4 shows the chart.
(4) Protein content 64.8%
(5) Sugar content 15.7%
(6) Composition ratio of main sugars Glucose: galactose: mannose = 1: 2: 1
(7) Amino acid composition Aspartic acid: 12.6%, glutamic acid: 13.2%, glycine 11.2%, alanine: 10.1%, valine: 6.7%, serine: 6.6%, leucine: 6 .7%.
実施例 3
実施例1または2で得た生理活性物質EEM−PNまたはPMを用いて粉末剤を調製し、下記のような抗癌試験を行った.抗癌試験における生存率を対照群と比較して抗癌活性を決定した.この結果を表1に示す.
(抗癌試験)
ルイス肺癌の生細胞をBDF1雌マウスの皮下に移植し、翌日よりそれぞれの実施例で得た生理活性物質EEM−PMまたはPNを1日1回500mg/kgを精製水に懸濁して20日間経口投与した.そして精製水だけを投与した対照群と茸の熱水抽出物投与群及び実施例1または2で得た生理活性物質を投与した群について、それぞれの群のマウスの平均生存日数から延命率を算出した結果を表1に示した.Example 3
A powder was prepared using the bioactive substance EEM-PN or PM obtained in Example 1 or 2, and the following anticancer test was performed. The anticancer activity was determined by comparing the survival rate in the anticancer test with that of the control group. The results are shown in Table 1.
(Anti-cancer test)
Live cells of Lewis lung cancer were transplanted subcutaneously into BDF1 female mice, and from the following day, the physiologically active substance EEM-PM or PN obtained in each example was suspended once a day in 500 mg / kg in purified water and orally administered for 20 days. It was administered. For the control group administered with purified water only, the group treated with the hot water extract of sputum and the group administered with the physiologically active substance obtained in Example 1 or 2, the survival rate was calculated from the average survival days of the mice in each group. The results are shown in Table 1.
本発明により、効率良く生理活性の強い物質が食用茸ナメコ、および/または、マイタケから簡単に得られ、薬品または健康食品として有用な製品が得られる. According to the present invention, a substance having high physiological activity can be easily obtained from edible sea cucumber and / or maitake, and a product useful as a medicine or health food can be obtained.
Claims (11)
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008050910A1 (en) * | 2006-10-27 | 2008-05-02 | Yukiguni Maitake Co., Ltd. | Grifola frondosa-origin substance having antiinfluenza virus activity and method of producing the same |
CN104225395A (en) * | 2013-07-27 | 2014-12-24 | 郭香荣 | Tumor-resistant traditional Chinese medicine and preparation method |
-
2003
- 2003-10-28 JP JP2003402253A patent/JP2005132810A/en active Pending
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008050910A1 (en) * | 2006-10-27 | 2008-05-02 | Yukiguni Maitake Co., Ltd. | Grifola frondosa-origin substance having antiinfluenza virus activity and method of producing the same |
JP2008106018A (en) * | 2006-10-27 | 2008-05-08 | Yukiguni Maitake Co Ltd | Substance having anti-influenza virus activity derived from grifola frondosa and its manufacturing method |
EP2087899A4 (en) * | 2006-10-27 | 2010-04-07 | Yukiguni Maitake Co Ltd | Grifola frondosa-origin substance having antiinfluenza virus activity and method of producing the same |
US8168196B2 (en) | 2006-10-27 | 2012-05-01 | Yukiguni Maitake Co., Ltd. | Grifola frondosa-derived substance having anti-influenza virus activity and method for producing the same |
CN104225395A (en) * | 2013-07-27 | 2014-12-24 | 郭香荣 | Tumor-resistant traditional Chinese medicine and preparation method |
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