JPS6356881B2 - - Google Patents
Info
- Publication number
- JPS6356881B2 JPS6356881B2 JP58086470A JP8647083A JPS6356881B2 JP S6356881 B2 JPS6356881 B2 JP S6356881B2 JP 58086470 A JP58086470 A JP 58086470A JP 8647083 A JP8647083 A JP 8647083A JP S6356881 B2 JPS6356881 B2 JP S6356881B2
- Authority
- JP
- Japan
- Prior art keywords
- general formula
- proteoglucan
- bonds
- main chain
- branched chains
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
- 240000001080 Grifola frondosa Species 0.000 claims description 15
- 235000007710 Grifola frondosa Nutrition 0.000 claims description 15
- 235000001674 Agaricus brunnescens Nutrition 0.000 claims description 13
- 229920001503 Glucan Polymers 0.000 claims description 10
- 239000002246 antineoplastic agent Substances 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 238000003809 water extraction Methods 0.000 claims description 7
- 239000004480 active ingredient Substances 0.000 claims description 5
- RJGDLRCDCYRQOQ-UHFFFAOYSA-N anthrone Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3CC2=C1 RJGDLRCDCYRQOQ-UHFFFAOYSA-N 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 3
- 239000003513 alkali Substances 0.000 claims description 3
- 239000000126 substance Substances 0.000 description 18
- 150000004676 glycans Chemical class 0.000 description 14
- 229920001282 polysaccharide Polymers 0.000 description 14
- 239000005017 polysaccharide Substances 0.000 description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 238000000034 method Methods 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 206010028980 Neoplasm Diseases 0.000 description 5
- 239000002244 precipitate Substances 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000002054 transplantation Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 230000001093 anti-cancer Effects 0.000 description 3
- 229940041181 antineoplastic drug Drugs 0.000 description 3
- 238000004817 gas chromatography Methods 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 102000003886 Glycoproteins Human genes 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000009702 cancer cell proliferation Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000002198 insoluble material Substances 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000002195 soluble material Substances 0.000 description 2
- DBTMGCOVALSLOR-DEVYUCJPSA-N (2s,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-3,5-dihydroxy-6-(hydroxymethyl)-4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-6-(hydroxymethyl)oxane-2,3,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](CO)O[C@H](O)[C@@H]2O)O)O[C@H](CO)[C@H]1O DBTMGCOVALSLOR-DEVYUCJPSA-N 0.000 description 1
- 229920002498 Beta-glucan Polymers 0.000 description 1
- 101710154757 Exo-beta-1,3-glucanase Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 229920001543 Laminarin Polymers 0.000 description 1
- 239000005717 Laminarin Substances 0.000 description 1
- 208000006268 Sarcoma 180 Diseases 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000002303 glucose derivatives Chemical class 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- WJLUBOLDZCQZEV-UHFFFAOYSA-M hexadecyl(trimethyl)azanium;hydroxide Chemical compound [OH-].CCCCCCCCCCCCCCCC[N+](C)(C)C WJLUBOLDZCQZEV-UHFFFAOYSA-M 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 230000007721 medicinal effect Effects 0.000 description 1
- 238000006140 methanolysis reaction Methods 0.000 description 1
- SLCVBVWXLSEKPL-UHFFFAOYSA-N neopentyl glycol Chemical compound OCC(C)(C)CO SLCVBVWXLSEKPL-UHFFFAOYSA-N 0.000 description 1
- 229940117969 neopentyl glycol Drugs 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 229910021332 silicide Inorganic materials 0.000 description 1
- FVBUAEGBCNSCDD-UHFFFAOYSA-N silicide(4-) Chemical compound [Si-4] FVBUAEGBCNSCDD-UHFFFAOYSA-N 0.000 description 1
- 229920002379 silicone rubber Polymers 0.000 description 1
- 239000004945 silicone rubber Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Description
〔産業上の利用分野〕
この発明は、サルノコシカケ科に属するマイタ
ケ（Grifola Frondosa）の子実体及び菌糸体を
熱水抽出して得たβ―１，６結合を主鎖とし、β
―１，３分枝鎖を高頻度に有する新規なプロテオ
グルカン及びそれを有効成分とする抗ガン剤に関
するものである。
〔従来の技術〕
従来から各種のキノコより抽出した多糖類が抗
ガン性を有することは知られているが、これらの
多糖類の化学構造は、主としてβ―１，３結合を
主鎖とし、これにβ―１，６分枝鎖が結合された
構造のグルカンである。
一方、キノコの中でもマイタケを原料としてそ
の中の多糖類を抽出する方法も、例えば特開昭58
―236395号において公知である。
〔発明が解決しようとする課題〕
特開昭58―236395号の発明は、マイタケを原料
とし、その菌糸体の培養液中に分泌された多糖類
を純度よく、安価かつ多量に製造することを目的
としたもので、これによつて得られた多糖類は、
アンスロン反応陽性、ニンヒドリン反応が陰性の
タンパク質を含まない多糖類であるが、得られた
多糖類の医薬としての有効性、特に抗ガン効果の
有無は明らかではない。
一方、β―１，３結合を主鎖とし、これにβ―
１，６分枝鎖が結合されたグルカンは、ガン等に
対する抗腫瘍剤としての薬効は認められている
が、さらに強力な抗ガン効果を有するものが日夜
研究されているのが現状である。
この発明の発明者等は、かゝる現状に鑑み、よ
り優れた薬理効果を有するキノコおよびその有効
成分を探索検討することを目的として、多種類の
キノコの中からマイタケを選定し、このマイタケ
の子実体について各種の抽出実験を繰り返し行つ
た結果、前記のグルカンとは化学構造の全く異な
つたプロテオグルカンを得、これが強い抗腫瘍性
を有することを見出してこの発明を完成するに至
つたのである。
〔課題を解決するための手段〕
すなわち、この発明の第１の発明は、マイタケ
を熱水抽出して得た、グルカン部が下記一般式(A)
又は(B)で表され、酸不溶、アルカリ可溶、アンス
ロン反応及びローリー反応が陽性な分子量200×
104のβ―１，６結合を主鎖とし、β―１，３分
枝鎖を高頻度に有することを特徴とするプロテオ
グルカンである。
一般式(A)
一般式(B)
（ただし、各式中の数字は位置を表し、ａ〜ｄ
は側鎖で、その位置は変わつてもよい。またｎは
20〜100の整数である。）
である。
また、第２の発明は、マイタケを熱水抽出して
得た、グルカン部が下記一般式(A)又は(B)で表さ
れ、酸不溶、アルカリ可溶、アンスロン反応及び
ローリー反応が陽性な分子量200×104のβ―１，
６結合を主鎖とし、β―１，３分枝鎖を高頻度に
有するプロテオグルカンを有効成分とすることを
特徴とする抗ガン剤である。
一般式(A)
一般式(B)
（ただし、各式中の数字は位置を表し、ａ〜ｄ
は側鎖で、その位置は変わつてもよい。またｎは
20〜100の整数である。）
である。
〔作用〕
この発明のプロテオグルカンは、サルノコシカ
ケ科に属するマイタケの子実体または菌糸体を熱
水処理して、後記する実施例１に記載した方法に
よつて前記一般式(A)および(B)で表されるβ―１，
６結合を主鎖としβ―１，３分枝鎖とする多糖類
として取得されるものである。
また、かゝる構造の多糖タンパク体は、ガスク
ロマトグラフによる分析等によつて容易に確認す
ることができるものである。
しかして、このような化学構造の多糖タンパク
体は、投与によつて比較的短期間に強いガン細胞
増殖抑制効果を示すと共に、毒性がなく、また投
与による副作用もない。
したがつて、前記β―１，３結合を主鎖とし、
β―１，６結合を分枝鎖が結合された構造の従来
のグルカンと比較して抗ガン剤として圧倒的優位
な作用を発揮することができるものである。
〔実施例〕
以下、実施例及び比較例を揚げてこの発明をよ
り具体的の説明する。
実施例 １
下記(1)〜(5)に従つてこの発明のプロテオグルカ
ンの製造と、これによつて得たプロテオグルカン
の化学構造を明らかにした。
(1) マイタケの子実体を粉砕し、500ｇの乾燥粉
末を得たのち、これに５の蒸留水を加え、
100℃で10時間の熱水抽出、また1.2気圧、温度
120℃で３時間オートクレーブで処理した。
冷却後遠心分離法によつて得た上澄液に等量
のエタノールを加え、温度４℃、20時間放置
後、14.5ｇの沈澱を集めた。
この沈澱１ｇに対し、水440mlを加えて溶解
し、PH12.0以上になるまでセチルトリメチルア
ンモニウムヒドロキシドを添加し、４℃で放置
後13.8ｇの沈澱物を得た。
このものを30％及び50％の酢酸で処理し、酸
可溶物を除いて３ｇの酸不溶物を得た。
この酸不溶物に６％水酸化ナトリウムを加え
てアルカリ可溶物を集め、これに４倍量の99％
エタノールを加えて沈澱を得た。
この物質にクロロホルム―メタノール混液
（２：１）を加え、はげしく撹拌してタンパク
を除いた。
生じた水層を分取し、これにエタノールを４
倍量添加後、温度４℃で20時間放置し、遠心分
離により約１ｇの沈澱を得た。
この物質は冷水に難溶な淡褐色粉末で、アン
スロン反応は陽性であり、ローリー（Lowry）
法により約22％のタンパク質が検出されたこと
から、プロテオグルカンであることが確認され
た。
(2) つぎに、この糖タンパク質をまずセフアロー
ス（sepharose）CL―4B，2.5×45cmカラムを
用いてゲル濾過し、分子量200×104の高分子糖
タンパク質を得た。
さらに、この物質をDEAE―セフアロース
CL―6B2.0×4.5cmカラムに吸着させ、PH7.2の
１／１５Ｍリン酸緩衝液で溶出される物質を得た。
この物質をセルロース透析チユーブを用いて
４℃で精製水に対して透析し、さらにセフアデ
ツクスＧ―25.3×37cmカラムで完全に脱塩し
た。
これを蒸発乾固させ、淡褐色無定形粉末状の
目的物を得た。
この物質は0.6％のタンパク質を含む多糖で
あつた。
(3) つぎに、前記(2)で得た物質の化学構造を明ら
かにした。
すなわち、前記の物質１mgを採り、５％塩酸
―メタノール１mlに溶解し、温度100℃で６時
間封管分解した後、分解物にトルエン−エタノ
ール（１：１）混液を添加して、減圧下で蒸発
乾固させた。
これにトリメチルけい素化剤を添加して温度
70℃で10分間反応させ、SE―20（シリコンゴム
GE）を充鎮剤とするガスクロマトグラフ分析
にかけたところ、グルコースのみが検出され
た。
さらに、この物質をエクソーβ―１，３―グ
ルカナーゼをPH5.0でマツキユルビン緩衝液の
下で反応させたところ、48時間後に構成グルコ
ース残基の約45％が遊離した。
このことから、この物質はβ―１，３結合を
有するグルカンであることが確認された。
(4) つづいて、この物質を箱守法により完全メチ
ル化した後、この完全メチル化物を５％塩酸―
メタノールによつてメタノリシスした。
これをネオペンチルグルコールサクシネート
（Neopentylglycol succinate）を充填剤とする
ガスクロマトグラフ分析したところ、β―１，
３結合に由来する２，４，６―トリ―Ｏ―メチ
ル、β―１，６結合に由来する２，３，６―ト
リ―Ｏ−メチル、１，３，６結合に由来する
２，４―ダイ―Ｏ―メチル及び非還元末端に由
来する２，３，４，６―テトラ―Ｏ―メチル―
グルコース誘導体の存在が認められた。なお、
そのモル比は２：１：４：４であつた。
(5) つぎに、得られた物質をスミス分解処理した
ところ、分子量200×104であつたこの物質は、
分子量３×104に低下した。
以上の結果から、マイタケ子実体を熱水抽出し
て得られたこの物質は、前記の一般式(A)又は(B)で
示されるβ―１，６結合と主鎖とし、これに高頻
度のβ―１，３分枝鎖を有する化学的構成よりな
る多糖タンパクであると結論された。
実施例２，３及び比較例１〜３
ICR5週令雄性マウス腋下にザルコーマ180固型
腫瘍を１×106細胞移植し、移植後に実施例１で
得たこの発明のプロテオグルカンを抗ガン剤とし
て経口投与した。
この投与は、投与量が１mg／Kg／ｄの場合（実
施例２）と、10mg／Kg／ｐの場合（実施例３）に
それぞれ分けて行い、いずれも細胞移植後１日目
及び９日目より各々10日間連続して行つたのち、
投与の最終日、すなわち投与の10日目に腫瘍重量
（ｇ）を測定した。
その結果を第１表に示す。
また、実施例２，３と併行して、蒸留水を与え
た対照例として比較例１、キノコより抽出して得
た抗ガン剤クレスチンPS―Ｋ（呉羽化学(株)製）の
所定量を投与した比較例２、及びβ―１，３結合
を主鎖として、β―１，６を分枝鎖とした抗ガン
剤ラミナリン（β―１，３ｇ lucan）の所定量
を投与した比較例３によつて同様の試験を実施し
たが、その結果も第１表に併記した。
なお、第１表上欄の投与法〔〕は細胞移植後
１日目より10日連続して腹腔内に注入したもので
あり、投与法〔〕は細胞移植後９日目より10日
間連続して腹腔内に注入したもので、各欄のＴ／
Ｃ℃は対照例（比較例１）の腫瘍重量を基準とし
てそれぞれの腫瘍重量の割合を示したものであ
る。
[Industrial Application Field] This invention uses β-1,6 bonds as the main chain obtained by hot water extraction of the fruiting body and mycelium of Maitake (Grifola Frondosa), which belongs to the family Aridaceae, and
This invention relates to a novel proteoglucan having a high frequency of -1,3 branched chains and an anticancer drug containing the same as an active ingredient. [Prior Art] It has been known that polysaccharides extracted from various mushrooms have anticancer properties, but the chemical structure of these polysaccharides is mainly composed of β-1,3 bonds as the main chain, This is a glucan with a structure in which β-1,6 branched chains are bonded to this. On the other hand, among mushrooms, there is also a method of extracting polysaccharides from maitake as a raw material, for example, in JP-A-58
- known in No. 236395. [Problem to be solved by the invention] The invention of JP-A No. 58-236395 is to produce polysaccharides secreted into the culture solution of mycelium using maitake mushrooms as a raw material, with high purity, at low cost, and in large quantities. The polysaccharide obtained by this method is
Although it is a protein-free polysaccharide that is positive for the Anthrone reaction and negative for the ninhydrin reaction, it is not clear whether the obtained polysaccharide is effective as a medicine, especially whether it has an anticancer effect. On the other hand, β-1,3 bonds are used as the main chain, and β-
Glucans with 1,6-branched chains have been recognized to have medicinal effects as antitumor agents against cancer, etc., but at present, products with even stronger anticancer effects are being researched day and night. In view of the current situation, the inventors of this invention selected Maitake mushrooms from among many types of mushrooms with the aim of searching for and investigating mushrooms with superior pharmacological effects and their active ingredients. As a result of repeated various extraction experiments on the fruiting bodies of the above-mentioned proteoglucan, they obtained a proteoglucan with a completely different chemical structure from the glucans mentioned above, and discovered that this had strong antitumor properties, leading to the completion of this invention. be. [Means for Solving the Problems] That is, the first invention of the present invention provides a glucan moiety obtained by hot water extraction of maitake mushrooms having the following general formula (A).
Or (B), acid insoluble, alkali soluble, positive for Anthrone reaction and Lowry reaction, molecular weight 200×
It is a proteoglucan characterized by having 10 4 β-1,6 bonds in its main chain and frequently having β-1,3 branched chains. General formula (A) General formula (B) (However, the numbers in each formula represent the positions, a to d
is a side chain whose position may vary. Also, n is
An integer between 20 and 100. ). Further, the second invention is a glucan moiety obtained by hot water extraction of maitake mushrooms, which is represented by the following general formula (A) or (B), is acid insoluble, alkali soluble, and has positive Anthrone reaction and Lowry reaction. β-1 with molecular weight 200×10 4 ,
It is an anticancer agent characterized by containing as an active ingredient proteoglucan, which has 6-bonds as its main chain and frequently has β-1,3 branched chains. General formula (A) General formula (B) (However, the numbers in each formula represent the positions, a to d
is a side chain whose position may vary. Also, n is
An integer between 20 and 100. ). [Effect] The proteoglucan of the present invention can be obtained by treating the fruiting body or mycelium of Maitake belonging to the family Arunocycolaceae with hot water, and by the method described in Example 1 to be described later. β-1,
It is obtained as a polysaccharide with 6 bonds as the main chain and β-1,3 branched chains. Further, polysaccharide proteins having such a structure can be easily confirmed by analysis using gas chromatography. Therefore, a polysaccharide protein having such a chemical structure exhibits a strong cancer cell proliferation suppressive effect in a relatively short period of time when administered, is not toxic, and has no side effects due to administration. Therefore, with the β-1,3 bond as the main chain,
Compared to conventional glucan, which has a structure in which β-1,6 bonds are linked to branched chains, it can exhibit overwhelmingly superior effects as an anticancer agent. [Example] The present invention will now be described in more detail with reference to Examples and Comparative Examples. Example 1 The proteoglucan of the present invention was produced and the chemical structure of the proteoglucan thus obtained was clarified according to the following (1) to (5). (1) Grind the fruiting body of maitake to obtain 500g of dry powder, add distilled water from step 5 to this,
Hot water extraction for 10 hours at 100℃, also 1.2 atm, temperature
It was autoclaved at 120°C for 3 hours. After cooling, an equal amount of ethanol was added to the supernatant obtained by centrifugation, and after standing at a temperature of 4°C for 20 hours, 14.5 g of precipitate was collected. To 1 g of this precipitate, 440 ml of water was added to dissolve it, and cetyltrimethylammonium hydroxide was added until the pH reached 12.0 or above, and after standing at 4° C., 13.8 g of precipitate was obtained. This product was treated with 30% and 50% acetic acid to remove acid-soluble materials to obtain 3 g of acid-insoluble materials. Add 6% sodium hydroxide to this acid-insoluble material, collect the alkali-soluble material, and add 4 times the amount of 99%
Ethanol was added to obtain a precipitate. A chloroform-methanol mixture (2:1) was added to this material and the protein was removed by vigorous stirring. Separate the resulting aqueous layer and add 4 ethanol to it.
After adding double the amount, it was left to stand at a temperature of 4°C for 20 hours, and about 1 g of precipitate was obtained by centrifugation. This substance is a light brown powder that is sparingly soluble in cold water, and the Anthrone reaction is positive.
Approximately 22% protein was detected using this method, confirming that it was proteoglucan. (2) Next, this glycoprotein was first subjected to gel filtration using a Sepharose CL-4B, 2.5 x 45 cm column to obtain a high molecular weight glycoprotein with a molecular weight of 200 x 104 . In addition, this substance is DEAE-cepharose.
A substance was obtained which was adsorbed onto a CL-6B 2.0 x 4.5 cm column and eluted with 1/15M phosphate buffer at pH 7.2. This material was dialyzed against purified water at 4° C. using a cellulose dialysis tube and completely desalted using a Sephadex G-25.3×37 cm column. This was evaporated to dryness to obtain the target product in the form of a pale brown amorphous powder. This material was a polysaccharide containing 0.6% protein. (3) Next, we clarified the chemical structure of the substance obtained in (2) above. That is, 1 mg of the above substance was taken, dissolved in 1 ml of 5% hydrochloric acid and methanol, and decomposed in a sealed tube at a temperature of 100°C for 6 hours. A mixture of toluene and ethanol (1:1) was added to the decomposed product, and the mixture was dissolved under reduced pressure. It was evaporated to dryness. Add trimethyl silicide to this and
After reacting at 70℃ for 10 minutes, SE-20 (silicone rubber)
When subjected to gas chromatography analysis using GE) as a packing agent, only glucose was detected. Furthermore, when this substance was reacted with exo-β-1,3-glucanase at pH 5.0 under Matsuki Urbin buffer, about 45% of the constituent glucose residues were liberated after 48 hours. From this, it was confirmed that this substance was a glucan having β-1,3 bonds. (4) Next, this substance was completely methylated by the Hakomori method, and then this completely methylated product was mixed with 5% hydrochloric acid.
Methanolysis was carried out with methanol. When this was analyzed by gas chromatography using neopentylglycol succinate as a packing material, β-1,
2,4,6-tri-O-methyl derived from 3 bonds, 2,3,6-tri-O-methyl derived from β-1,6 bonds, 2,4 derived from 1,3,6 bonds -Di-O-methyl and 2,3,4,6-tetra-O-methyl derived from the non-reducing end-
The presence of glucose derivatives was observed. In addition,
The molar ratio was 2:1:4:4. (5) Next, when the obtained substance was subjected to Smith decomposition treatment, this substance with a molecular weight of 200×10 4 was
The molecular weight decreased to 3×10 4 . From the above results, this substance obtained by hot water extraction of maitake fruiting bodies has β-1,6 bonds and main chains represented by the above general formula (A) or (B), and has a high frequency of It was concluded that it is a polysaccharide protein with a chemical composition having β-1,3 branched chains. Examples 2 and 3 and Comparative Examples 1 to 3 1×10 6 cells of Sarcoma 180 solid tumor were transplanted into the armpit of a 5-week-old male mouse with ICR, and after the transplantation, the proteoglucan of the present invention obtained in Example 1 was administered as an anticancer agent. Orally administered as This administration was carried out separately when the dose was 1 mg/Kg/d (Example 2) and when the dose was 10 mg/Kg/p (Example 3), and both were administered on the 1st and 9th day after cell transplantation. After doing this for 10 consecutive days,
Tumor weight (g) was measured on the last day of administration, ie, the 10th day of administration. The results are shown in Table 1. In addition, in parallel with Examples 2 and 3, as a control example in which distilled water was given, Comparative Example 1, a predetermined amount of the anticancer agent Crestin PS-K (manufactured by Kureha Chemical Co., Ltd.) extracted from mushrooms was added. Comparative Example 2, in which a predetermined amount of the anticancer drug laminarin (β-1,3g lucan), which has β-1,3 bonds as the main chain and β-1,6 branched chains, was administered in Comparative Example 3. A similar test was carried out by , and the results are also listed in Table 1. The administration method [ ] in the upper column of Table 1 is intraperitoneal injection for 10 consecutive days from the 1st day after cell transplantation, and the administration method [ ] is for 10 consecutive days from 9th day after cell transplantation. Injected intraperitoneally, T/ in each column
C°C indicates the ratio of each tumor weight based on the tumor weight of the control example (Comparative Example 1).
以上詳述したとおり、この発明はサルノコシカ
ケ科に属するマイタケの子実体を熱水処理して、
得られる前記一般式(A)または(B)で表されるβ―
１，６結合を主鎖としβ―１，３分枝鎖とする多
糖タンパク体からなるプロテオグルカンおよびこ
のプロテオグルカンを有効成分として含有させた
抗ガン剤である。
かゝる化学構造のプロテオグルカンは、抗ガン
剤として従来のβ―１，３結合を主鎖としβ―
１，６分枝鎖とする多糖よりなるグルカンに比べ
てそのガン細胞増殖抑制効果が顕著に優れてお
り、食用とされているマイタケより得られる抽出
物であるため、毒性の影響を何等受けることがな
く、きわめて有用性の高いものである。
さらに、近年特に安価に人工裁培されている前
記のマイタケを原料として抽出処理によつて得ら
れるものであるから、廉価かつ容易に取得するこ
とができるものであり、これたの点でもこの発明
は大きな価値を有するものである。
As detailed above, this invention involves hot water treatment of the fruiting body of maitake, which belongs to the family Salmonaceae.
β- represented by the above general formula (A) or (B) obtained
This product is a proteoglucan consisting of a polysaccharide protein having a main chain of 1,6 bonds and β-1,3 branched chains, and an anticancer agent containing this proteoglucan as an active ingredient. Proteoglucan with such a chemical structure has a β-1,3 bond as the main chain and is used as an anti-cancer drug.
Compared to glucans made of 1,6-branched polysaccharides, it has a significantly superior cancer cell proliferation inhibitory effect, and since it is an extract obtained from maitake mushrooms, which are considered edible, it is not affected by toxicity in any way. It is extremely useful. Furthermore, since it is obtained by extraction treatment using the above-mentioned maitake mushroom, which has been artificially cultivated at a particularly low cost in recent years, it can be obtained inexpensively and easily. is of great value.
Claims (1)
下記一般式(A)又は(B)で表され、酸不溶、アルカリ
可溶、アンスロン反応及びローリー反応が陽性な
分子量200×104のβ―１，６結合を主鎖とし、β
―１，３分枝鎖を高頻度に有することを特徴とす
るプロテオグルカン。 一般式(A) 一般式(B) （ただし、各式中の数字は位置を表し、ａ〜ｄ
は側鎖で、その位置は変わつてもよい。またｎは
20〜100の整数である。） ２ マイタケを熱水抽出して得た、グルカン部が
下記一般式(A)又は(B)で表され、酸不溶、アルカリ
可溶、アンスロン反応及びローリー反応が陽性な
分子量200×104のβ―１，６結合を主鎖とし、β
―１，３分枝鎖を高頻度に有するプロテオグルカ
ンを有効成分とすることを特徴とする抗ガン剤。 一般式(A) 一般式(B) （ただし、各式中の数字は位置を表し、ａ〜ｄ
は側鎖で、その位置は変わつてもよい。またｎは
20〜100の整数である。）[Claims] 1 Molecular weight obtained by hot water extraction of maitake mushrooms, whose glucan moiety is represented by the following general formula (A) or (B), is acid-insoluble, alkali-soluble, and has a positive Anthrone reaction and Lowry reaction. The main chain consists of 200×10 4 β-1,6 bonds, and β
- A proteoglucan characterized by having a high frequency of 1,3 branched chains. General formula (A) General formula (B) (However, the numbers in each formula represent the positions, a to d
is a side chain whose position may vary. Also, n is
An integer between 20 and 100. ) 2 Glucan moiety obtained by hot water extraction of maitake mushrooms, whose glucan moiety is represented by the following general formula (A) or (B), is acid insoluble, alkali soluble, and has a molecular weight of 200 β-1,6 bond is the main chain, β
-An anticancer agent characterized by containing as an active ingredient a proteoglucan having a high frequency of 1,3-branched chains. General formula (A) General formula (B) (However, the numbers in each formula represent the positions, a to d
is a side chain whose position may vary. Also, n is
An integer between 20 and 100. )
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP58086470A JPS59210901A (en) | 1983-05-17 | 1983-05-17 | Glucan having beta-1,6 bond-containing main chain, obtained from maitake and antineoplastic agent comprising same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP58086470A JPS59210901A (en) | 1983-05-17 | 1983-05-17 | Glucan having beta-1,6 bond-containing main chain, obtained from maitake and antineoplastic agent comprising same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS59210901A JPS59210901A (en) | 1984-11-29 |
| JPS6356881B2 true JPS6356881B2 (en) | 1988-11-09 |
Family
ID=13887842
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP58086470A Granted JPS59210901A (en) | 1983-05-17 | 1983-05-17 | Glucan having beta-1,6 bond-containing main chain, obtained from maitake and antineoplastic agent comprising same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS59210901A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0500718A1 (en) * | 1989-11-09 | 1992-09-02 | Byron A Donzis | Insoluble yeast extract. |
| WO2009102008A1 (en) * | 2008-02-14 | 2009-08-20 | Yukiguni Maitake Co., Ltd. | Low-molecular-weight substance derived from maitake mushroom and having immunostimulating activity and anti-tumor activity |
Families Citing this family (24)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6058925A (en) * | 1983-09-09 | 1985-04-05 | Nippon Beet Sugar Mfg Co Ltd | Antineoplastic substance |
| JPS60255733A (en) * | 1984-05-30 | 1985-12-17 | Nippon Beet Sugar Mfg Co Ltd | Beta-d-glucan |
| US5622939A (en) * | 1992-08-21 | 1997-04-22 | Alpha-Beta Technology, Inc. | Glucan preparation |
| WO1991003248A2 (en) * | 1989-09-08 | 1991-03-21 | Alpha Beta Technology, Inc. | Method for immune system activation |
| US5488040A (en) * | 1989-09-08 | 1996-01-30 | Alpha-Beta Technology, Inc. | Use of neutral soluble glucan preparations to stimulate platelet production |
| US5811542A (en) * | 1989-09-08 | 1998-09-22 | Alpha-Beta Technology, Inc. | Method for producing soluble glucans |
| EP0490995A1 (en) * | 1989-09-08 | 1992-06-24 | Alpha Beta Technology | Method for producing soluble glucans |
| JP2689244B2 (en) * | 1993-04-30 | 1997-12-10 | 株式会社雪国まいたけ | Method for producing liver disease improving agent |
| JP2753935B2 (en) * | 1993-04-30 | 1998-05-20 | 株式会社雪国まいたけ | Method for producing immunosuppressant |
| JP2732008B2 (en) * | 1993-04-30 | 1998-03-25 | 株式会社雪国まいたけ | Method for producing hair growth promoter |
| US5622940A (en) * | 1994-07-14 | 1997-04-22 | Alpha-Beta Technology | Inhibition of infection-stimulated oral tissue destruction by β(1,3)-glucan |
| JPH08119874A (en) * | 1994-10-24 | 1996-05-14 | Takumi Sogabe | Production of suppressor of aids virus and cancer cell |
| AUPN166195A0 (en) | 1995-03-13 | 1995-04-06 | Norvet Research Pty Limited | Process for glucan extraction |
| JP2859843B2 (en) * | 1996-03-08 | 1999-02-24 | 株式会社雪国まいたけ | Antitumor substance extracted from Maitake |
| US6046323A (en) * | 1997-07-29 | 2000-04-04 | The Collaborative Group, Ltd. | Conformations of PPG-glucan |
| US6616928B1 (en) | 1998-10-20 | 2003-09-09 | Yukiguni Maitake Co., Ltd. | Active oxygen scavenger and cancer chemopreventer from Grifola |
| AU6261999A (en) | 1998-09-25 | 2000-04-17 | Collaborative Group, Ltd., The | Very high molecular weight beta-glucans |
| US6369216B1 (en) | 1998-09-25 | 2002-04-09 | Biopolymer Engineering Pharmaceutical, Inc. | Very high molecular weight β-glucans |
| JP2001097881A (en) * | 1999-09-28 | 2001-04-10 | Yukiguni Maitake Co Ltd | Cancer prophylaxis agent, and cancer prophylaxis food or feed all derived from grifola frondosa |
| JP2005133069A (en) * | 2003-10-09 | 2005-05-26 | Ichimasa Kamaboko Co Ltd | METHOD FOR ACQUISITION OF MUSHROOM-DERIVED beta-GLUCAN POLYSACCHARIDE AND MUSHROOM-DERIVED BETA-GLUCAN POLYSACCHARIDE |
| JP5153188B2 (en) * | 2007-03-30 | 2013-02-27 | 小林製薬株式会社 | Th1 / Th2 balance improver |
| JP2011184306A (en) * | 2010-03-04 | 2011-09-22 | Yukiguni Maitake Co Ltd | Substance derived from grifola spp. mushroom suppressing elevation of postprandial hyperglycemia |
| CN103304680B (en) * | 2012-03-09 | 2017-02-08 | 中国科学院上海药物研究所 | Beta-glucan, and extraction method and application thereof |
| CN104497161B (en) * | 2015-01-04 | 2016-09-14 | 华东理工大学 | A kind of method of purification of grifolan |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5836395A (en) * | 1981-08-26 | 1983-03-03 | Nippon Beet Sugar Mfg Co Ltd | Preparation of polysaccharide |
-
1983
- 1983-05-17 JP JP58086470A patent/JPS59210901A/en active Granted
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5836395A (en) * | 1981-08-26 | 1983-03-03 | Nippon Beet Sugar Mfg Co Ltd | Preparation of polysaccharide |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0500718A1 (en) * | 1989-11-09 | 1992-09-02 | Byron A Donzis | Insoluble yeast extract. |
| EP0500718A4 (en) * | 1989-11-09 | 1994-09-14 | Byron A Donzis | Insoluble yeast extract |
| WO2009102008A1 (en) * | 2008-02-14 | 2009-08-20 | Yukiguni Maitake Co., Ltd. | Low-molecular-weight substance derived from maitake mushroom and having immunostimulating activity and anti-tumor activity |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS59210901A (en) | 1984-11-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JPS6356881B2 (en) | ||
| EP1027061B1 (en) | Imine-forming polysaccharides, preparation thereof and the use thereof as adjuvants and immunostimulants | |
| US4454315A (en) | Carboxymethylated derivatives of β-1,3-glucan | |
| JPH08259602A (en) | New polysaccharide substance | |
| US3987166A (en) | Treatment of tumors with glucan compositions in mice and rats | |
| US3893996A (en) | Hydroxyalkyl and mercaptoalkyl derivatives of {62 -(1{43 3) glucans | |
| JPH06340701A (en) | Highly branched beta-glucan, its production and use | |
| HU176828B (en) | Process for producing nitrogen-containing polysaccharides of tumor-inhibiting activity | |
| KR870001930B1 (en) | Preparation method of polysaccharides,ron | |
| JPH026761B2 (en) | ||
| US4973581A (en) | Glucan derivatives having tumoricidal activity | |
| JP2630783B2 (en) | Method for producing neutral polysaccharide having antitumor activity | |
| JPH0248000B2 (en) | ||
| US5185327A (en) | Glucan derivatives having tumoricidal activity | |
| JPS6377901A (en) | Novel polysaccharides | |
| EP0173228A2 (en) | Polysaccharide rin substance, production of the same and use of the same | |
| JPH06312934A (en) | Production of substance having immunosuppressive effect | |
| JPH0645645B2 (en) | Chemically modified polysaccharide and method for producing the same | |
| JPS5993092A (en) | Protein polysaccharide | |
| JPS63307825A (en) | Antitumor agent and production thereof | |
| JPH0645644B2 (en) | Chemically modified polysaccharide and method for producing the same | |
| KR19980058723A (en) | New water-soluble polysaccharides with antitumor activity and methods for their preparation | |
| JPS60155202A (en) | Chemically modified polysaccharide and its production | |
| CA1094453A (en) | TREATMENT OF TUMORS AND NEW DERIVATIVES OF .beta.-1,3- GLUCAN | |
| EP0267335B1 (en) | Glucan derivatives |