200412989 (1) 玖、發明說明 【發明所屬之技術領域】 本發明爲提供一種含香菇菌絲體萃取物，r 6 τ細胞 的活性增強劑之開發，即，關於免疫活性劑的開發以及提 供。 本發明更係含香菇菌絲體萃取物，r <5 τ細胞的活性 增強作用，即，關於提供具有免疫活性作用的食品、飲料 以及飼料的開發。 本發明進一步地爲關於提供含香菇菌絲體萃取物，抗 腫瘤劑、抗細菌感染治療劑以及抗病毒感染治療劑之開發 【先前技術】 r 5 τ細胞的特徵 於動物的血液中存在的末梢τ細胞，由其細胞表面抗 原的丁細胞受容體（T C R )的種類，大略分爲二種類之 τ細胞，其存在係公知的事。其中一種爲於細胞表面具有 T C R α鏈以及/3鏈的α /9 丁細胞，另一種爲T C R 7鏈 以及具有5鏈7 (5 τ細胞。對於人類，r 6 τ細胞對一般 末梢血液以及淋巴組織僅存在數％至1 0 %程度，其爲完 全與α /3 T細胞具有相異特徵的傷害殺傷細胞。 7 <5 Τ細胞存在於人的腸管、皮膚以及末梢血液中等 ’作用於局部免疫。作爲r 6 τ細胞的功能，有對癌細胞 傷害活性、細菌或病毒的感染防禦活性等至今被報告出。 (2) (2)200412989 7 5 T細胞的感染防禦活性 脾臟等被認定的r 5 τ細胞中，存在著感染時產生 I L 一 4或I FN-α等cytokine的細胞。在實驗上作出 不存在這些細胞的狀況時，已知會對細菌感染抵抗性減弱 。例如，有報告指出，對投與7 6型T C R抗體時可過渡 性地抑制r 6 T細胞機能的老鼠或τ c R 6基因缺失老鼠 而言，可降低 Mycobacterium tuberculosis的感染之抵抗 性（Ladel C ? e t al .，Ευ r J Immunol, 1995,25:2877-2881 ). 。又有報告指出Listeria monocytogenes的感染的初期有 7 (5T 細胞出現（Hiromatsu K，et al.，J Exp Med, 1992, 175:4 9- 5 6 )。由這些可知，r (5 T細胞對.於細菌感染爲 生物體防禦的重要細胞。 且，有報告指出B型肝炎病毒的慢性感染，肝臟以及 月卑臟的7 5 T細胞會被誘發增殖（Ο z a k i S ; e t a ] · 5 J M e d I nveSi; 1 9 9 8,44:2 1 5 -2 17)。又，與正常老鼠比較，缺欠r 5 T 細胞的老鼠體內，對於牛痘病毒感染初期，有報告指出會有 顯著地增力D (Welsh RM，et al .，Immunol Rev， 1 997, 1 5 9 : 7 9 - 9 3 )。可此可知，7 (5 T細胞不僅對於上述細菌 感染症有作用，對於病毒的感染亦顯出有作用。 7 (5 T細胞的癌細胞傷害活性 r 5 T細胞係對自身癌細胞有著特異殺傷性，且係對 於自身的正常淋巴球（例如a /3 T細胞）卻完全無顯示殺 -6 - (3) (3)200412989 傷的T細胞。此點表示，使用活性7 (5 T細胞對癌作治療 時，有著極少產生副作用的危險性之效果。與r 6 τ細胞 相反的a /3 T細胞因已知其對自身癌細胞無殺傷，而會殺 傷自身白血球，對於癌的治療因活性化a /3 Τ細胞而有著 產生嚴重副作用的危險性。因此，對癌的治療考慮使用活 性化的7 6 Τ細胞爲可期待的。 且7 6 Τ細胞爲Μ H C非限制性地殺死癌細胞，有著 與Ν Κ細胞類似的性質。r (5 Τ細胞於幼兒末梢血液中約 有1 0 %存在著，但隨著年齡的增加其數會減少。因此有 報告指出隨著年齡的增長癌的發生便提高其爲原因，此與 體內的7 (5 T細胞減少有關。又，雞、綿羊、牛等其末稍 血中1 5 %至5 0 %者有r (5 T細胞存在。由這些動物中 較少發生癌症的事看來，顯示末梢r (5 τ細胞的存在與癌 症的抑制有著密切的關係。 香菇的藥理效果 香菇（Leutinus edodes )於日本、中國爲代表性的食 用菇，日本約3 0 0年前便進行人工栽培。日常食用的菇 類爲子實體，其爲菌類爲使下一代能留下而產生胞子的生 殖體，係爲營養體的菌絲細胞於地中或原木中經常時間的 生長。 香菇自古便有傳說對各種疾病或症狀有治療效果，但 對其藥理作用的解明僅爲最近的事。對於香菇菌絲體萃取 物的小白鼠、家鼠之引發癌症實驗中，可抑制動物的大腸 -7- (4) (4)200412989 、肝臟等腫瘤形成或移植腫瘤細胞的增殖，提升動物的生 存率(N . S u g a η 〇 e t al.，Cancer Letter, 27:1，1985;鈴 木康將們，日本大腸肛門病會誌，43 :1 7 8,1 990等），顯示促進 細胞分裂活性（T . Tabata et al . ? Im mu nopharmacology, 24:5 7， 1 9 9 2; Y . Hibino et al . ，Immunopharmacology, 2 8:7 7, 1 9 94 等）、增強抗體的產生，藉著抗體 ADCC(antibody-dependentcell-mediated cytotoxicty)顯示 免疫學上對肝細胞障礙的抑制效果（溝口靖紘們，肝膽塍 ，1 5 : 1 2 7，1 9 8 7 )等種種有報告。 由此見解可知香菇成分的藥理成份之硏究，已進行至 醫學藥學的分野中。其結果，香菇成份的一部分可使用於 人類免疫能力回復進而對癌症等之治療，又已知可能具有 抑制引發癌症之事。 本發明的目的爲，對香菇菌絲體萃取物的藥理作用更 詳細說明，則爲探索香菇菌絲體萃取物的新穎性醫藥、食 品、飮料、飼料等用途。 本發明的目的爲開發、提供一種含有香菇菌絲體萃取 物的r δ τ細胞活性增強物質，並且又開發、提供一種免 疫活性劑，例如爲抗腫瘤劑、抗細菌感染治療劑以及抗病 毒感染治療劑。 本發明另一目的爲，使用含有香菇菌絲體萃取物的 7 δ Τ細胞活性增強物質，或者使用含有香菇菌絲體萃取 物的免疫活性劑，治療被檢驗體的腫瘤。 (5) (5)200412989 【發明內容】 〔發明的揭示〕 本發明者爲解決上述課題而g羊細硏究結果’發現香链 菌絲體萃取物顯示有顯著增強r 6 T細胞活性的作用，便 完成本發明。 即，本發明的目的爲開發、提供一種含有香蘇菌絲體 萃取物的7 (5 T細胞活性增強物質’並且又開發、提供一 種免疫活性劑，例如爲抗腫瘤劑、抗細菌感染治療劑以及 抗病毒感染治療劑。 本發明的7 5 T細胞活性增強物質或免疫活性劑係， 可含有香薛菌絲體萃取物以及作爲任一成份之藥劑上可容 許的載體，以醫藥組成物的型態。 又，本發明的7 5 τ細胞活性增強物質或免疫活性劑 ，可經口投與用、注射用、經黏膜投與用、經消化管投與 用或經皮投與用任一種皆可。 又，本發明的7 6 Τ細胞活性增強物質或免疫活性劑 可以食品、飮料、或飼料任一種型態皆可。 【實施方式】 〔實施發明的最佳型態〕 使用於本發明的7 (5 Τ細胞活性增強的香菇菌絲體萃 取物，係由香菇菌於固體培養基下培養所得的菌絲體，較 佳爲含有菌絲體的固體培養基，於水與酵素存在下粉碎， 分解所得的萃取物而言。 -9- (6) (6)200412989 香菇菌絲體萃取物較佳爲使用以下的方法所得者，但 不限定於此。即’蔗渣（甘蔗渣）與脫脂米糠爲基材的固 體培養基上接種香菇菌，再將其菌絲體增殖所得的含有菌 絲體的固體培養基通過1 2篩孔（每平方英吋孔眼數）的 份量爲3 0重量％以下而解束。此被解束的固體培養基中 添加水以及選自於纖維酶、蛋白酶或葡萄糖酶的一種或以 上的酵素，前述固體培養基保持3 0至5 5 °C的溫度而添 加的同時，前述的固體培養基於前述酵素存在下粉碎，禱 爛使蔗渣最少以7 0重量％以上的份量通過1 2篩孔。再 加熱至9 5 °C的溫度使其酵素失去活性而同時滅菌，所得 的懸濁狀液經濾過而得到香菇菌絲體萃取物。此香菇菌絲 體萃取物可直接使用於本發明的7 6 T細胞活性增強物質 ，亦可將此濃縮、冷凍乾燥成爲粉末保存，使用時可因種 種的型態而便利使用。冷凍乾燥所得的粉末爲褐色粉末， 具吸濕性，且具特異味道與臭味。 香薛菌絲體萃取物的體內（i n v i v 〇 )中7 T細胞活 性增強物質以下述的實施例所記載的方法進行試驗時，觀 察到顯著的7 (5 T細胞活性增強效果。 本發明的7 5 T細胞活性增強物質對因7 (5 T細胞對 腫瘤細胞進行傷害而對腫瘤之治療以及/或預防具有效果 。本發明的7 5 T細胞活性增強物質並無對特定的腫瘤細 胞有特性的治療，其特徵爲以提高r 6 T細胞的活性，因 7 <5 T細胞的活性化其結果可以破壞腫瘤細胞爲目的。即 ，使用本發明的7 (5 T細胞活性增強物質對作爲治療對象 -10- (7) (7)200412989 的腫瘤細胞之治療，不僅爲惡性腫瘤細胞，良性腫瘤細胞 即可，不限定特定腫瘤細胞。因此，含有香菇菌絲體萃取 物以及作爲任意成份而藥劑上可容許的載體之r δ τ細胞 活性增強物質，可作爲任一種腫瘤治療用以及/或預防用 組成物而使用。 本發明的r (5 τ細胞活性增強物質，可作爲細菌感染 或病毒感染的治療用以及/或預防用組成物使用。本發明 的7 5 τ細胞活性增強物質非對特定細菌或病毒有者特異 的作用，而以提高7 (5 T細胞的活性，以其結果被感染細 菌或病毒由體內排除爲目的。本發明的7 (5 T細胞活性增 強物質可治療的細菌性或病毒性疾病，例如有結梭菌（ Mycobacterium sp ·)、單核白血球增多性李士德菌 (Listeria monocytogenes)、肝炎病毒（A 型、B 型、C 型） 、人類免疫不全病毒（Human Immunodeficiency Virus) > 牛痘病毒（Vaccinia virus)等可舉出，但不侷限於此。 作爲治療用以及/或預防用組成物的投與路線，雖經 口投與最佳，但以靜脈內投與、腹腔內投與、皮下投與、 肌肉內投與、經鼻投與、經皮投等亦可。作爲經口投與較 佳的製劑含有，錠劑、膠囊、散劑、顆粒劑、溶液劑、糖 漿劑等，但不侷限於此。經鼻投與或經皮投與較適合的製 劑含有貼劑、貼布劑等，但不侷限於此。 藥劑可容許的載體含有斯業界公知的適當賦形劑、分 解劑、滑潤劑、香料、著色劑、溶解補助劑、懸濁劑、塗 布劑等。但不侷限於此。 -11 - (8) (8)200412989 作爲本發明的L A K活性增強用製劑可任意混合的藥 學上容許之載體，斯業界中公知的賦形劑（例如有乳糖、 葡萄糖、澱粉、結晶纖維等）、結合劑（澱粉、明膠、甲 基纖維、聚乙烯比各烷酮等）、分解劑（澱粉、羧基甲基 纖維鈣、羧基甲基澱粉等）、潤滑劑（滑石、硬脂酸鹽等 )、塗布劑（白糖、滑石、明膠等）等，且各種光澤化劑 、香料、著色劑、矯味劑、溶解補助劑、安定化劑、懸濁 劑、吸收促進劑等可因應目的而添加，但不侷限於此。又 ，作爲注射劑使用時，對於此分野可使用一般慣用的各種 稀釋劑（例如有水、乙醇等）。 本發明的r 5 τ細胞活性增強物質的投與量依被檢驗 者的年齡、體重、症狀、投與途徑等被考慮而由醫師決定 。本發明的7 5 T細胞活性增強物質所含有的香菇菌絲體 萃取物原本即可作爲食品使用，由其爲極爲安全的事看來 ’雖無需嚴格限定其投與量，但香廷菌絲體萃取物1日數 次（2至3次左右），1次量爲1〇0 m g至1〇〇 〇〇 m g ( 1曰的總投與量約爲2〇〇m g至3 0 0〇0 m g )爲佳’且較佳爲1曰3次，1日約5〇0 m g至 5 0 0 0 m g (總投與量爲1日約1 5 0 0 m g至 15000 mg)，且更佳爲1曰3次，1日約100〇 m g至1 5 0 〇 m g (總投與量爲1日約3 〇 0 0 m g至 4 5 0 0 m g )。且，可與其他抗腫瘤劑合倂使用。 本發明的r (5 τ細胞活性增強細胞物質爲，提供一種 治療腫瘤時於被動免疫法所使用的製劑。作爲被動免疫法 -12- (9) (9)200412989 ，於被感染的細胞中一般將淋巴球移入生物體內，達到殺 死腫瘤細胞爲目的的腫瘤治療方法。本發明的情況，首先 將來自被檢驗體的的末梢血分離7 5 T細胞，分離後的 r δ τ細胞於體外中以本發明的r (5 τ細胞活性增強物質 活性化後，活性化後的r (5 τ細胞再次還回被檢驗體的體 內。結果，因活性化的r (5 τ細胞的作用，將可破壞體內 的腫瘤細胞。 本發明的r 6 τ細胞活性增強物質，可直接使用香菇 菌絲體萃取物亦可，或含有香菇菌絲體萃取物的r 5 τ細 胞活性增強物質，以及含有藥學上許可的載體之醫藥，或 獸醫藥用組成物即可。 本發明的7 δ T細胞活性增強物質可用食品的型態提 供。較佳的食品型態爲粉末、顆粒、糊狀、膠狀等可舉出 。且使用顆粒等時，作爲可添加的甘味料，可添加乳糖等 糖類。又，本發明的7 (5 丁細胞活性增強物質可以飮料的 型態提供。如此之食品或飮料中，含有特定保健用食品、 病者用食品等範圍。 本發明的7 5 Τ細胞活性增強物質，做爲飼料或添加 於飼料中添加劑的形式提供亦可。作爲家畜的飼料或飼料 中的添加劑’因使用本發明的7 (5 Τ細胞活性增強物質， 可治療以及/或預防家畜發生腫瘤，或可對於家畜的細菌 或病毒性感染症之治療以及/或預防。其結果，對於家畜 現今所使用的治療藥，例如可減少抗生素物質等的使用量 ，並可減低隨之的飼料成本。且，具有可降低因抗生物質 -13- (10) (10)200412989 投與時生產物無法出貨的時間之驚人效果。 對於人類被檢驗體，體內中的r (5 τ細胞活性增強效 果1 1式驗法如以下進行。首先將香菇菌絲萃取物粉末3 . 6 g經7天每天接種於人類被檢驗體（全量爲2 5 . 2 g ) °再以香菇菌絲體萃取物服用後的末梢血中7 3 T細胞頻 $與服用前的當該細胞頻率以使用流動細胞測試儀測定並 作比較。 本發明雖於下述的實施例中更詳細說明，但其不能限 定本發明的範圍。本發明的方法可由斯業者作種種變更、 修飾而使用，這些則包含於本發明中。 實施例 實施例1 :香菇菌絲體萃取物的調製法 蔗渣9 0重量份，米糠1 0重量份所成的固體培養基 使其含適當水後，接種香菇種菌，放置於調整溫度以及濕 度的培養室內，使菌絲體增殖。菌絲體於固體培養基下聚 集後，解開蔗渣基材的纖維素束，使其成爲1 2篩孔（每 平方英吋的孔眼數）可通過份量爲2 4重量％以下。此解 束之後的培養基1 · 0 k g中，純水3 · 5 L以及純化的 纖維酵素2 · 0 g ’以邊保持固體培養基爲4 〇 °c邊加入 ，成爲含有培養基的混合物。 其次’含有培養基的混合物以附有變速齒輪的幫浦一 邊將其循環’一邊將固體培養基於齒輪部分進行粉碎以及 揆爛作用2 0 0分鐘的程度’使其蔗渣纖維能約以8 〇重 -14- (11) (11)200412989 量％作爲1 2篩孔的通過份量，含有培養基的混合物進行 粉碎與揆爛爲，邊徐徐上升該混合物的溫度邊實施。其後 ’含有培養基的混合物再加溫至9 0 °C放置3 〇分鐘。使 酵素失去活性同時進行滅菌，於9 0 °C下放置3 0分鐘。 因9 0 °C的加熱使得酵素失去活性，且實施殺菌過程。所 得的含有培養基混合液以6 0篩孔濾布過濾作爲香菇菌絲 體萃取物，將此濃縮後經凍結乾燥，得到香菇菌絲萃取物 原始粉末。 如此所得的香菇菌絲體萃取物以苯酚硫酸法進行糖値 分析得知含有糖値爲2 5 · 3 % (重量/重量）、以洛瑞 法（Lowry法）進行蛋白質分析得知含有蛋白質.i 9 . 7 % (重量/重量），以沒食子酸爲基準的佛隆-丹尼斯法 (Fol on-Denis法）得知含有聚苯酚2 · 6 % (重量/重 量）。晉菇菌絲體萃取物中亦含有其他粗脂肪8 % 、粗灰 份2 2 % ，糖質以外的可溶性無氨物約2 0 % 。200412989 (1) 发明. Description of the invention [Technical field to which the invention belongs] The present invention is to provide an activity enhancer for r 6 τ cell-containing extract of lentinus edodes mycelium extract, that is, the development and provision of an immunologically active agent. The present invention further relates to a mushroom mycelium-containing extract, and r < 5 τ cell activity-enhancing effect, that is, the development of providing food, beverage, and feed having an immune-active effect. The present invention is further related to the development of providing extracts of mycelia containing shiitake mushrooms, antitumor agents, antibacterial infection treatment agents, and antiviral infection treatment agents. [Previous technology] r 5 τ cells are characterized by the presence of peripheral blood in animals Tau cells are roughly divided into two types of tau cells by the type of T cell receptor (TCR) on their cell surface antigens, and their existence is well known. One type is α / 9 D cells with TCR α chain and / 3 chain on the surface of the cell, and the other is TCR 7 chain and 5 chain 7 (5 τ cell. For humans, r 6 τ cells are more common for peripheral blood and lymph. Tissues exist only to a few% to 10%, which is a killer killer cell with completely different characteristics from α / 3 T cells. 7 < 5 T cells are present in the human intestines, skin and peripheral blood, etc. Immune. As a function of r 6 τ cells, noxious activity against cancer cells, infection defense activity against bacteria or viruses, etc. have been reported so far. (2) (2) 200412989 7 5 T cell infection defense activity is recognized Among r 5 τ cells, there are cells that produce cytokine such as IL-4 or I FN-α upon infection. It is known that resistance to bacterial infection is weakened when experimental conditions are made in the absence of these cells. For example, reports indicate that For mice that can transiently inhibit r 6 T cell function or mice with τ c R 6 gene deletion when administered with a TCR antibody of type 7 6, it can reduce the resistance to Mycobacterium tuberculosis infection (Ladel C? Et al Ευ r J Immunol, 1995, 25: 2877-2881). It was also reported that 7 (5T cells appeared in the early stage of infection of Listeria monocytogenes (Hiromatsu K, et al., J Exp Med, 1992, 175: 4 9- 5 6). From these, r (5 T cell pairs are important cells for biological defense against bacterial infection. In addition, there are reports of chronic infection with hepatitis B virus, liver and 7 5 T Cells will be induced to proliferate (〇 zaki S; eta] · 5 JM ed InveSi; 1 9 9 8,44: 2 1 5 -2 17). Compared with normal mice, mice lacking r 5 T cells, In the early stage of vaccinia virus infection, there have been reports of a significant increase in D (Welsh RM, et al., Immunol Rev, 1 997, 1 5 9: 7 9-9 3). It can be seen that 7 (5 T cells Not only has an effect on the above bacterial infections, but also has an effect on virus infection. 7 (5 T cell cancer cell damaging activity r 5 T cell line has specific killing properties to its own cancer cells, and it is also its own normal lymph Spheres (such as a / 3 T cells) are completely absent from killing -6-(3) (3) 200412989 injured T cells. This point table It has been shown that the use of active 7.5 T cells for the treatment of cancer has very little risk of side effects. As opposed to r 6 τ cells, a / 3 T cells are known to have no killing effect on their own cancer cells, but will kill their own white blood cells. There is a risk of serious side effects in the treatment of cancer by activating a / 3 T cells. Therefore, considering the use of activated 76 T cells for cancer treatment is expected. And 7 6 T cells are M H C kill cancer cells without limitation, and have similar properties to N K cells. r (5 T cells are present in about 10% of young children's peripheral blood, but their number will decrease with age. Therefore, there are reports that the incidence of cancer increases with age. This is related to the body It is related to the decrease of 7 (5 T cells. In addition, 15 to 50% of the terminal blood of chickens, sheep, cattle, etc. have r (5 T cells). It seems that less cancer occurs in these animals. It shows that the presence of peripheral r (5 τ cells) is closely related to the suppression of cancer. The pharmacological effects of shiitake mushrooms (Leutinus edodes) are representative edible mushrooms in Japan and China. Artificial cultivation was carried out in Japan about 300 years ago. The daily edible mushrooms are the fruiting bodies, which are the germ cells of the fungi that produce the spores for the next generation to leave, and the mycelial cells, which are vegetative, have been growing in the ground or logs for a long time. It is said to have a therapeutic effect on various diseases or symptoms, but the explanation of its pharmacological effects is only recent. It has been shown to inhibit the animal's large intestine in mice and house mice induced by lentinus mycelium extract. (4) (4) 200412989 Formation of tumors such as liver and liver or the proliferation of transplanted tumor cells to improve the survival rate of animals (N. Suga eta et al., Cancer Letter, 27: 1, 1985; General Suzuki, Japan Journal of Colorectal and Anal Diseases, 43 : 1 7 8, 1 990, etc.), showing cell division promoting activity (T. Tabata et al.? Im mu nopharmacology, 24: 5 7, 1 9 9 2; Y. Hibino et al., Immunopharmacology, 2 8: 7 7, 1 9 94, etc.), enhance the production of antibodies, and demonstrate immunologically inhibitory effects on hepatocyte disorders through antibody ADCC (antibody-dependentcell-mediated cytotoxicty) (Mizoguchi Yasugi, Hepatobiliary, 1 5: 1 2 7 There are reports from various sources such as 19 8 7). From this knowledge, we can know that the research on the pharmacological ingredients of the mushroom ingredients has been carried out in the field of medical pharmacy. As a result, a part of the mushroom ingredients can be used to restore human immunity and thus to cancer. It is also known that other treatments may have the effect of inhibiting the initiation of cancer. The purpose of the present invention is to explain the pharmacological effects of the shiitake mushroom mycelium extract in more detail, and to explore the novel medicine and food of the shiitake mushroom mycelium extract Material The purpose of the present invention is to develop and provide an r δ τ cell activity-enhancing substance containing a mycelium extract of Lentinus edodes, and to develop and provide an immunoactive agent such as an antitumor agent and antibacterial infection treatment. Agent and antiviral infection treatment agent. Another object of the present invention is to use a 7 δ T cell activity-enhancing substance containing an extract of Lentinus edodes mycelium, or to use an immunoactive agent containing an extract of Lentinus edodes mycelium to treat the test subject Tumor. (5) (5) 200412989 [Summary of the invention] [Disclosure of the invention] In order to solve the above-mentioned problems, the present inventors have studied the results carefully and found that the extract of Streptomyces mycelium showed a significant effect of enhancing r 6 T cell activity. This completes the present invention. That is, the object of the present invention is to develop and provide a 7 (5 T cell activity enhancing substance) containing an extract of Basil mycelium, and to develop and provide an immunoactive agent such as an antitumor agent and an antibacterial infection treatment agent. And a therapeutic agent for anti-viral infection. The 7 5 T cell activity-enhancing substance or immune-active agent system of the present invention may contain the extract of Chancel mycelium and a pharmaceutically acceptable carrier as any component, and a pharmaceutical composition In addition, the 7 5 τ cell activity-enhancing substance or immunological agent of the present invention can be used for oral administration, injection, transmucosal administration, digestive tract administration, or transdermal administration. In addition, the 7 6 T cell activity-enhancing substance or immunoactive agent of the present invention may be in any form of food, food, or feed. [Embodiment] [The best form for carrying out the invention] Use in the present invention 7 (5 T cell activity-enhanced lentinus mycelium extract is mycelium obtained by culturing lentinus in a solid medium, preferably a solid medium containing mycelium, which is stored in water and enzymes. For the extract obtained by pulverizing and decomposing below. -9- (6) (6) 200412989 The mushroom mycelium extract is preferably obtained by the following method, but is not limited thereto. That is, 'bagasse (bagasse) The solid medium containing defatted rice bran as a substrate was inoculated with Lentinus edodes, and then the mycelia-producing solid medium containing the mycelium was passed through 12 sieve openings (eyes per square inch), and the weight was 30% by weight. The disintegration is performed in the following manner. Water and one or more enzymes selected from the group consisting of cellulase, protease, and glucosease are added to the disintegrated solid medium, and the solid medium is added while maintaining a temperature of 30 to 55 ° C. The aforementioned solid medium was pulverized in the presence of the aforementioned enzymes, and prayed to rot the bagasse through at least 70% by weight of the sieve through 12 sieve. Then heated to a temperature of 95 ° C to deactivate the enzyme and sterilize it simultaneously. The suspension-like liquid is filtered to obtain a shiitake mushroom mycelium extract. The shiitake mushroom mycelium extract can be directly used in the 7 6 T cell activity enhancing substance of the present invention, or it can be concentrated and freeze-dried to a powder It can be stored and used conveniently due to various types. The powder obtained by freeze-drying is a brown powder, hygroscopic, and has a specific taste and odor. In vivo (inviv 〇) 7 T cell activity-enhancing substances When tested by the method described in the following Examples, a significant 7 (5 T cell activity-enhancing effect was observed. The 7 5 T cell activity-enhancing substance of the present invention has a factor of 7 (5 T cell pairs). The tumor cells are injured to have an effect on the treatment and / or prevention of tumors. The 7 5 T cell activity enhancing substance of the present invention has no specific treatment for specific tumor cells, and is characterized by increasing r 6 T cell activity, The purpose of activating 7 < 5 T cells is to destroy tumor cells as a result. That is, the treatment of the tumor cells to be treated with the 7 (5 T cell activity enhancing substance of the present invention -10- (7) (7) 200412989) is not limited to malignant tumor cells and benign tumor cells, and the specific tumor is not limited. Therefore, an r δ τ cell activity-enhancing substance containing an extract of shiitake mushroom mycelium and an pharmaceutically acceptable carrier as an arbitrary component can be used as any composition for tumor treatment and / or prevention. The r (5 τ cell activity enhancing substance can be used as a composition for the treatment and / or prevention of bacterial or viral infections. The 7 5 τ cell activity enhancing substance of the present invention does not have a specific effect on specific bacteria or viruses In order to increase the activity of 7 (5 T cells), the purpose is to exclude infected bacteria or viruses from the body. The 7 (5 T cell activity enhancing substance) of the present invention can treat bacterial or viral diseases, such as Clostridium (Mycobacterium sp.), Listeria monocytogenes, Hepatitis virus (types A, B, C), human immune system Examples include but are not limited to Human Immunodeficiency Virus & Vaccinia virus etc. The route of administration of the therapeutic and / or prophylactic composition is best though oral administration, but Intravenous administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, nasal administration, transdermal administration, etc. are also acceptable. As a preferred preparation for oral administration, tablets, capsules, powders, Granules, solutions, syrups, etc. are not limited to this. Suitable preparations for nasal or transdermal administration include, but are not limited to, patches. Permissible carriers include Suitable excipients, disintegrating agents, lubricants, perfumes, colorants, dissolution aids, suspending agents, coating agents, etc. are well known in the industry. However, it is not limited to this. -11-(8) (8) 200412989 as the present The formulation for enhancing the LAK activity of the present invention can be arbitrarily mixed with a pharmaceutically acceptable carrier, excipients (such as lactose, glucose, starch, crystalline fiber, etc.) and binders (starch, gelatin, methyl fiber, Polyethylene than alkanone , Decomposing agents (starch, carboxymethyl cellulose calcium, carboxymethyl starch, etc.), lubricants (talc, stearate, etc.), coating agents (sugar, talc, gelatin, etc.), and various glossing agents, spices , Colorants, flavoring agents, dissolution aids, stabilizers, suspending agents, absorption enhancers, etc. can be added according to the purpose, but it is not limited to this. In addition, when used as an injection, generally used for this field Various diluents (for example, water, ethanol, etc.) The dose of the r 5 τ cell activity-enhancing substance of the present invention is determined by the physician depending on the age, weight, symptoms, and route of administration of the test subject. The shiitake mushroom mycelium extract contained in the 7 5 T cell activity enhancing substance of the present invention can be used as a food originally. From the point of view of being extremely safe, 'though it is not necessary to strictly limit the dosage, Shantin hypha Body extract several times a day (about 2 to 3 times), the amount of each time is 100 mg to 10,000 mg (the total amount of administration is about 2000 mg to 3 000 mg) is better 'and is preferably 3 times a day, about 500 mg to 5000 mg per day (total dosage is about 15 000 mg to 15000 mg per day), and more preferably Three times a day, about 100 mg to 1 500 mg per day (the total dose is about 300 mg to 4 500 mg per day). In addition, it can be used in combination with other antitumor agents. The r (5 τ cell activity enhancing cell material) of the present invention is to provide a preparation for passive immune method for treating tumors. As a passive immune method-12- (9) (9) 200412989, it is generally used in infected cells. A tumor treatment method for transferring lymphocytes into a living body to achieve the purpose of killing tumor cells. In the case of the present invention, peripheral blood from a test subject is firstly separated into 7 5 T cells, and the isolated r δ τ cells are in vitro. After activation of the r (5 τ cell activity-enhancing substance of the present invention, the activated r (5 τ cells are returned to the body of the test subject again. As a result, the effect of the activated r (5 τ cells will be Destroy tumor cells in the body. The r 6 τ cell activity-enhancing substance of the present invention can be used directly from the shiitake mushroom mycelium extract, or the r 5 τ cell activity-enhancing substance containing the shiitake mushroom mycelium extract, and contains pharmaceutical The approved carrier medicine or veterinary medicinal composition can be used. The 7 δ T cell activity enhancing substance of the present invention can be provided in the form of food. The preferred food form is powder, granule, paste, gum Examples include the shape and the like. When granules are used, sugars such as lactose can be added as a sweetener. In addition, the 7-cell activity enhancing substance of the present invention can be provided in the form of a raw material. Such a food or a raw material It contains a range of foods for specific health care, foods for patients, etc. The 7 5 T cell activity-enhancing substance of the present invention can also be provided as a feed or an additive added to feed. It can also be used as a feed for livestock or an additive in feed. 'The use of the 7T cell activity-enhancing substance of the present invention can treat and / or prevent tumors in livestock, or can treat and / or prevent bacterial or viral infections in livestock. As a result, for livestock today, The therapeutic drugs used, for example, can reduce the amount of antibiotic substances used, and can reduce the associated feed costs. Moreover, it can reduce the production of antibiotics due to anti-biomass 13- (10) (10) 200412989 The amazing effect of time. For human subjects, the r (5 τ cell activity enhancement effect in the body of the test method 11 is performed as follows. First, the mushroom mycelium is extracted 3.6 g of powder was inoculated into human subjects every day for 7 days (the total amount was 25.2 g) ° 7 3 T cells in peripheral blood after taking shiitake mushroom mycelium extract were more frequent than before The cell frequency is measured and compared using a flow cell tester. Although the present invention is described in more detail in the following examples, it cannot limit the scope of the present invention. The method of the present invention can be changed and modified by industry practitioners. These are used in the present invention. Examples Example 1: Preparation method of 90 parts by weight of bagasse and 10 parts by weight of rice bran in a solid culture medium made of a suitable medium containing water, and then inoculating the mushrooms The inoculum is placed in an incubation chamber with temperature and humidity adjusted to proliferate the mycelium. After the mycelium was aggregated in a solid medium, the cellulose bundles of the bagasse substrate were unraveled to make it into 12 sieve openings (the number of perforations per square inch), and the passing amount was 24% by weight or less. In this 1.0-g medium after disintegration, 3.5-L of pure water and 2.0-g of purified cellulase were added while maintaining the solid medium at 40 ° C to form a mixture containing the medium. Next, 'the mixture containing the culture medium was circulated by a pump with a gear shifting gear' while pulverizing and rotting the solid culture medium to the gear part to the extent of 200 minutes', so that the bagasse fiber could be about 80 weight- 14- (11) (11) 200412989 The amount% is a passing amount of 12 sieve holes. The mixture containing the culture medium is pulverized and rotted, and the temperature of the mixture is gradually increased while the temperature is increased. Thereafter, the mixture containing the culture medium was further warmed to 90 ° C and left for 30 minutes. The enzyme was deactivated and sterilized, and left at 90 ° C for 30 minutes. Enzymes are deactivated by heating at 90 ° C and a sterilization process is performed. The obtained mixed liquid containing the culture medium was filtered through a 60-mesh filter cloth as an extract of lentinus mycelium, which was concentrated and freeze-dried to obtain an original powder of lentinus mycelium extract. The thus obtained lentinus mycelium extract was analyzed for glycocalyx by the phenol sulfuric acid method and found that the content of glycocalyx was 25.3% (w / w), and the protein was analyzed by the Lowry method (Lowry method). i 9.7% (w / w), based on the Fol on-Denis method based on gallic acid, was found to contain 2.6% (w / w) of polyphenol. Mushroom mycelium extract also contains 8% of other crude fats, 22% of crude ash, and about 20% of soluble ammonia-free substances other than sugar.
其中香菇菌絲體萃取物中所構成的糖組成物如以下所 示：x y 1 : 1 5 · 2 ; A a : 8 · 2 ; M a η : 8-4；Gul：39.4；Gal：5.4；GlcN • 12-0;GLuUa :11.3。 貫施例2 ··杳菇菌絲體萃取物的7 5 T細胞活性增強體內 試驗 人類被檢驗者3名（被檢驗者Α〜C )以香菇菌絲體 萃取物原始粉末3 . 6 g /日經7天每天經口服用（全量 -15- (12) (12)200412989 爲2 5 · 2 g )。經過香菇菌絲體萃取物的服用期間後， 自人類被檢驗體取出末梢血。服用後所取的末梢血中之 r 5 T細胞頻率，與服用前所取的末梢血中之細胞頻率作 比較，以流動細胞測試器測定。所得的結果如圖1以及圖 2所示。 3名全部人員皆與香菇菌絲體萃取物的服用前作比較 ，服用後的其末梢血中T (5 T細胞的比率平均上升4 0 % 以上。 表1 :對服用前後，末梢血r T細胞比率的增加率The sugar composition formed in the extract of Lentinus edodes mycelium is as follows: xy 1: 1 5 · 2; A a: 8 · 2; M a η: 8-4; Gul: 39.4; Gal: 5.4; GlcN • 12-0; GLuUa: 11.3. Example 2 · 7 5 T cell activity enhancement of Mycelium edodes mycelium extract in vivo test 3 human subjects (subjects A to C) with lentinus mycelium extract original powder 3.6 g / For daily use of Nikkei for 7 days (full amount -15- (12) (12) 200412989 is 2 5 · 2 g). After the administration period of the shiitake mushroom mycelium extract, the peripheral blood is removed from the human subject. The frequency of r 5 T cells in peripheral blood taken after taking was compared with the frequency of cells in peripheral blood taken before taking, and was measured with a flow cell tester. The obtained results are shown in Figs. 1 and 2. All 3 persons were compared with before the administration of Lentinus edodes mycelium extract, and the T (5 T cell ratio in peripheral blood increased by an average of more than 40% after taking it. Table 1: Peripheral blood T cells before and after administration Rate of increase
被檢驗者 被檢驗者 被檢驗者 平均±SEM A B C 增力D 1 2 4.3 9% 146.15% 1 50.00% 1 4 0 . 1 8 % ± 7.7 9 % 率 另一方面，對由同一被檢驗者中取出服用前後的末梢 血液，對r 5 τ細胞以外的標示物作調查。其結果，香菇 菌絲體萃取物中未見到對7 δ τ細胞的增値活性，其平均 値卻顯示細胞比率的減少（圖3 )。 產業上可利用性 含有本發明的香菇菌絲體萃取物之r (5 τ細胞活性增 強物質，得知實際上可活性化7 (5 T細胞。由此可知，本 發明的r 5 T細胞活性增強物質因會誘導具有7 6 T細胞 -16- (13) (13)200412989 的腫瘤細胞傷害活性、抗細菌感染以及抗病毒感染作用， 而具有生物體對腫瘤、細菌感染以及病毒感染的防禦效果 ，可使用於腫瘤、細菌感染以及病毒感染的預防、治療。 又，r τ細胞對本發明的r (5 τ細胞活性增強物質，因 無副作用故可安全地使用，期待可利用於龐大產業上。 又，對於引起細菌感染以及/或病毒感染的家畜而言 ，現今所使用的治療藥，例如抗生物質等可減少其使用量 ，隨之可降低飼養成本。且，因不使用抗生物質，故可減 短生產品無法出貨的時間之如此進一步效果。 【圖式簡單說明.】 圖1爲表示，服甩本發明的香菇菌絲體萃取物後， 7 (5 T細胞於末梢血中比率的提高。 圖2爲表示，服用香菇菌絲體萃取物的前後，7 5 丁 細胞的流動位置檢測解析（Π 〇 w s i t e m e t r y )之，結果；。 圖3爲表示，服用香‘薛菌絲體萃取物後α点T細胞於 末梢血中的比率，換言之爲因減少而作爲照圖表示。 -17-Testees Testees Testees average ± SEM ABC increase D 1 2 4.3 9% 146.15% 1 50.00% 1 4 0. 1 8% ± 7.7 9% Peripheral blood before and after the administration was examined for markers other than r 5 τ cells. As a result, no increase activity of 7 δ τ cells was observed in the extract of Lentinus edodes mycelium, but the average 平均 showed a decrease in the cell ratio (FIG. 3). Industrial availability The r (5 τ cell activity enhancing substance containing the shiitake mushroom mycelium extract of the present invention was found to actually activate 7 (5 T cells. From this, it can be seen that the r 5 T cell activity of the present invention The enhancement substance can induce tumor cell nociceptive activity, antibacterial infection and antiviral infection with 7 6 T cells-16- (13) (13) 200412989, and has the defense effect of the organism against tumor, bacterial infection and viral infection It can be used for the prevention and treatment of tumors, bacterial infections, and viral infections. Furthermore, r τ cells can safely be used for the r (5 τ cell activity-enhancing substance of the present invention without side effects, so it can be used in a large industry. In addition, for domestic animals that cause bacterial and / or viral infections, currently used therapeutic drugs, such as antibiotics, can reduce the amount used, and can reduce the cost of breeding. Moreover, because antibiotics are not used, they can be used. Such a further effect of shortening the time during which raw products cannot be shipped. [Brief description of the drawing.] Figure 1 shows that after taking the shiitake mushroom mycelium extract of the present invention, 7 ( The increase in the ratio of 5 T cells to peripheral blood. Figure 2 shows the results of detection and analysis of the flow position of the 7 5 D cells before and after taking the mycelium extract of Lentinula edodes, and the results are shown in Figure 3. The ratio of α-point T cells in peripheral blood after taking Xiang'Xue mycelium extract, in other words, it is shown as a picture because it is reduced. -17-