JP4928060B2 - イザリア型虫草由来のサイトカイン産生増強剤 - Google Patents
イザリア型虫草由来のサイトカイン産生増強剤 Download PDFInfo
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- JP4928060B2 JP4928060B2 JP2003336435A JP2003336435A JP4928060B2 JP 4928060 B2 JP4928060 B2 JP 4928060B2 JP 2003336435 A JP2003336435 A JP 2003336435A JP 2003336435 A JP2003336435 A JP 2003336435A JP 4928060 B2 JP4928060 B2 JP 4928060B2
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- isaria
- cytokine production
- interleukin
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Description
(1)天然の虫草菌を人工培養して発生するイザリア型虫草、その培養物もしくは抽出物、又はこれらの処理物を有効成分として含有するサイトカイン産生増強剤。
(2)サイトカインがインターロイキン、インターフェロン及びCSFから選ばれる少なくとも1種である前記(1)に記載のサイトカイン産生増強剤。
(3)インターロイキンがインターロイキン−2である前記(2)に記載のサイトカイン産生増強剤。
(4)インターロイキンがインターロイキン−6である前記(2)に記載のサイトカイン産生増強剤。
(5)インターロイキンがインターロイキン−10である前記(2)に記載のサイトカイン産生増強剤。
(6)インターフェロンがインターフェロンγである前記(2)に記載のサイトカイン産生増強剤。
(7)CSFがGM−CSFである前記(2)に記載のサイトカイン産生増強剤。
(8)イザリア型虫草の培養物又はその処理物を有効成分として含有する前記(1)〜(7)のいずれかに記載のサイトカイン産生増強剤。
(9)天然の虫草菌を人工培養して発生するイザリア型虫草の菌糸体、分生胞子又は二次代謝産物を有効成分として含有するサイトカイン産生増強剤。
(10)イザリア型虫草がハナサナギタケ(Isaria japonica Yasuda)又はツクツクボウシタケ(Isaria sinclairii (Berk.) Lloyd)である前記(1)〜(9)のいずれかに記載のサイトカイン産生増強剤。
(11)イザリア型虫草由来でサイトカイン産生増強活性を有する物質を有効成分として含有するサイトカイン産生増強剤。
(12)食品に添加するための前記(1)〜(11)のいずれかに記載のサイトカイン産生増強剤。
(13)前記(1)〜(11)のいずれかに記載のサイトカイン産生増強剤を有効成分として含有する、サイトカインの産生又は活性が低下した疾患の治療剤。
野外で発生する虫草菌の中には、子実体に子嚢を形成しないで胞子(分生子)が直接、子実体に付着する種類もあり、これを不完全世代型の虫草菌という。
子嚢菌亜門:ASCOMYTINA
核菌網:Pyrenomycetes
麦角菌科(肉座菌科):Claviciptaceae
ノムシタケ属:Cordyceps
(1)コルジセプス(Cordyceps = セミタケ、サナギタケ、冬虫夏草など)
(2)トルビエラ(Torrubiella= 節足動物クモに寄生のサンゴクモタケ、カイガラムシ寄生のカイガラムシタケ)
(3)ポドネクテリオデス(Podonectrioides= ヨコバイに寄生するヨコバイタケなど)
完全世代型の虫草菌は前記の3種の属型が多く、代表とされている。
(1)イザリア(Isaria = ハナサナギタケ、コナサナギタケ、セミに寄生するツクツクボウシタケなど)
(2)ギベルラ(Gibellula= 節足動物のクモに寄生するギベルラタケ)
(3)ヒメノスチベ(Hymenostilbe= トンボに寄生するヤンマタケなど)
(4)ポリセファロマイセス(Polycephalomyces= 鱗翅目の幼虫に寄生するマユダマタケなど)
不完全世代型はおおよそ前記の4種の属型に代表されるが、野外ではイザリア型(Isaria)の虫草菌が最も多い。
乾燥酵母0.6%(W/V)、植物タンパクのプロビアン(協和発酵)1.5%(W/V)、ペプトン0.2%(W/V)、寒天粉末2.2%(W/V)を蒸留水で溶解し、オートクレーブ滅菌し(120℃、20分)、40℃前後に冷却した段階で、メンブランフィルター(孔径0.2μm)で濾過滅菌し、グルコース1.5%(W/V)と、イノシン0.01%(W/V)水溶液を添加する。滅菌した培養液を高圧蒸気滅菌した三角フラスコ(100ml)に20mlずつ分注し、固化させて寒天培地を作成する。
イザリア型虫草の培養方法は、特に制限はなく、公知の方法、例えば、静置液体培養(例えば、特公昭61−53033号公報、特許第2746532号公報)を用いることができる。
この種の製剤には、適宜前記賦形剤の他に、結合剤、崩壊剤、界面活性剤、滑沢剤、流動性促進剤、矯味剤、着色剤、香料等を使用することができる。
また、本発明のサイトカイン産生増強剤及び治療剤は、液剤、シロップ剤、懸濁剤、乳剤、エリキシル剤として投与する場合には、矯味矯臭剤、着色剤を含有してもよい。
イザリア型虫草の一例であるハナサナギタケ(Isaria japonica Yasuda)又はツクツクボウシタケ(Isaria sinclairii (Berk.) Lloyd)の原菌をグルコース0.1〜2.0%、乾燥酵母(岩城製薬社)0.1〜2.0%、イノシン0.01〜0.1%、寒天1.0%の組成よりなる培地に接種し、試験管内で斜面、分離培養を行った。10〜30℃で14〜21日間培養した結果、得られた子実体は天然界のものと形態的、顕微鏡的のいずれにおいても同一であった。ここに得られた子実体を人口培養用の接種用原菌とした。
製造例1で得られたハナサナギタケ(Isaria japonica Yasuda)又はツクツクボウシタケ(Isaria sinclairii (Berk.) Lloyd)の培養物の凍結乾燥粉末を用い、消化管にある特殊なリンパ組織のパイエル板で実験を行った。
雄性のC57BL/6Jマウス(8〜9週齢)の小腸からパイエル板を摘出し、コラゲナーゼで結合組織を分解してからパイエル板構成リンパ球を採取した。次いで、パイエル板細胞を2×106個/mLになるように培地(5%ウシ胎児血清含有RPMI1640培地)に懸濁し、24穴プラスチックプレートにまいた。これに10又は100μg/mLの濃度になるように調製した前記ハナサナギタケ又はツクツクボウシタケの培養物の凍結乾燥粉末を添加し、更に5μg/mLの濃度のT細胞活性化タンパクであるコンカナバリンAを添加あるいは添加しないで96時間培養した。なお、パイエル板に存在するリンパ球は、T細胞を刺激する条件がなければサイトカイン産生を起こさないので、培養系にはコンカナバリンAを検体(ハナサナギタケ又はツクツクホウシタケの培養物)を添加するときと同時に添加し、サイトカインの産生を促す実験条件も加えた。
雄性C57BL/6Jマウス(8〜9週齢)に、生理食塩水のみ(コントロール群)、ハナサナギタケ培養物の凍結乾燥粉末(10mg/kg/日)、又はツクツクボウシタケ培養物の凍結乾燥粉末(10mg/kg/日)を7日間経口投与した後、小腸からパイエル板を摘出し、コラゲナーゼで結合組織を分解してからパイエル板構成リンパ球を採取した。次いで、パイエル板細胞を2×106個/mLになるように培地(5%ウシ胎児血清含有RPMI1640培地)に懸濁し、24穴プラスチックプレートにまいた。これに5μg/mLの濃度のT細胞活性化タンパクであるコンカナバリンAを添加あるいは添加しないで96時間培養した。
NS 培養系に何も添加しないで培養したときの値と対比して、有意な差が認められなかったことを示す。
Claims (14)
- ツクツクボウシタケ(Isaria sinclairii(Berk.) Lloyd)、その培養物もしくは抽出物、又はこれらの処理物を有効成分として含有し、インターロイキン−2、インターロイキン−6、インターロイキン−10、インターフェロンγ及びGM−CSFから選ばれるサイトカインのパイエル板構成細胞による産生を増強するためのサイトカイン産生増強剤。
- インターロイキン−2、インターロイキン−6、インターロイキン−10、インターフェロンγ及びGM−CSFのパイエル板構成細胞による産生を増強するために用いられる請求項1記載のサイトカイン産生増強剤。
- サイトカインがインターロイキン−2である請求項1記載のサイトカイン産生増強剤。
- サイトカインがインターロイキン−6である請求項1記載のサイトカイン産生増強剤。
- サイトカインがインターロイキン−10である請求項1記載のサイトカイン産生増強剤。
- サイトカインがインターフェロンγである請求項1記載のサイトカイン産生増強剤。
- ハナサナギタケ(Isaria japonica Yasuda)又はツクツクボウシタケ(Isaria sinclairii(Berk.) Lloyd)から選ばれるイザリア型虫草、その培養物もしくは抽出物、又はこれらの処理物を有効成分として含有し、GM−CSFのパイエル板構成細胞による産生を増強するためのサイトカイン産生増強剤。
- ハナサナギタケ(Isaria japonica Yasuda)又はツクツクボウシタケ(Isaria sinclairii (Berk.) Lloyd)から選ばれるイザリア型虫草の培養物又はその処理物を有効成分として含有する請求項7記載のサイトカイン産生増強剤。
- ハナサナギタケ(Isaria japonica Yasuda)又はツクツクボウシタケ(Isaria sinclairii (Berk.) Lloyd)から選ばれるイザリア型虫草の菌糸体、分生胞子又は二次代謝産物を有効成分として含有し、GM−CSFのパイエル板構成細胞による産生を増強するためのサイトカイン産生増強剤。
- イザリア型虫草がハナサナギタケ(Isaria japonica Yasuda)であり、経口投与用である請求項7〜9のいずれか1項に記載のサイトカイン産生増強剤。
- イザリア型虫草がツクツクボウシタケ(Isaria sinclairii(Berk.) Lloyd)である請求項7〜9のいずれか1項に記載のサイトカイン産生増強剤。
- ツクツクボウシタケ(Isaria sinclairii(Berk.) Lloyd)由来でサイトカイン産生増強活性を有する物質を有効成分として含有し、インターロイキン−2、インターロイキン−6、インターロイキン−10、インターフェロンγ及びGM−CSFから選ばれるサイトカインのパイエル板構成細胞による産生を増強するためのサイトカイン産生増強剤。
- 食品に添加するための請求項1〜12のいずれか1項に記載のサイトカイン産生増強剤。
- 請求項1〜12のいずれか1項に記載のサイトカイン産生増強剤を有効成分として含有する、サイトカインの産生又は活性が低下した疾患の治療剤。
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