經濟部中央標準局員工消費合作社印製 577745 __----- 五、發明説明（/ ) 發明領域 本發明關於一種具有抗癌活性及止痛活性之腫瘤血管 生成抑制劑，一種包括該腫瘤血管生成抑制劑之藥物組成 物及一種藉由使用該腫瘤血管生成抑制劑來治療癌症的方 法。 技藝背景 鯊魚是一種屬於軟骨魚網（Chondrichthyes)且出 現於近海之魚類，其骨骼爲軟骨，且已知鯊魚軟骨’其經 由捕獲和屠宰鯊魚及分離該軟骨而被製備，其具有抗腫瘤 活性且在癌症的治療上有效。例如，Car ti laid ’其爲一種 來自美國鯊魚之軟骨產品，在市場上作爲一種抗腫瘤物 質。 當生長至大約一至二立方毫米大小時，腫瘤產生一種 血管生成因子，因而腫瘤藉由其本身而能提供營養素及氧 作爲其生長所需。既然此新血管增生的抑制有助於腫瘤細 胞生長的抑制及在癌症治療上有效，許多尋找具有血管生 成抑制活性之物質的硏究正在進行中且已知鯊魚軟骨具有 此血管生成抑制活性。 然而，鯊魚軟骨對血管生成抑制之有效每天劑量爲5 0至6 0公克之大量且其味道及氣味令人厭惡及，因此’ 其口服施藥在許多實例上是困難的。既然鯊魚軟骨是一種 黏多糖的混合物，除了口服的途徑外，並沒有其他的施藥 途徑可用且所需的是，在口服施藥的時候，在強酸的狀況 下，鯊魚軟骨在胃中保持不溶，但在吸收位置，其溶解於 ______3___ 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） -- (請先閲讀背面之注意事項再填寫本頁)Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 577745 __----- V. Description of the Invention (/) Field of the Invention The present invention relates to a tumor angiogenesis inhibitor with anticancer activity and analgesic activity, and a tumor angiogenesis including the tumor angiogenesis Inhibitor pharmaceutical composition and a method for treating cancer by using the tumor angiogenesis inhibitor. Technical background Shark is a fish belonging to the Chondrichthyes and found offshore, its bones are cartilage, and shark cartilage is known to be prepared by capturing and slaughtering sharks and isolating the cartilage. It has antitumor activity and is It is effective in treating cancer. For example, Car ti laid ’is a cartilage product from American sharks, which is used as an anti-tumor substance in the market. When it grows to about one to two cubic millimeters in size, the tumor produces an angiogenic factor, so the tumor itself can provide nutrients and oxygen as its growth needs. Since this inhibition of neovascularization contributes to the inhibition of tumor cell growth and is effective in the treatment of cancer, many investigations for finding substances with angiogenesis inhibitory activity are ongoing and it is known that shark cartilage has this angiogenesis inhibitory activity. However, the effective daily dose of shark cartilage to inhibit angiogenesis is a large amount of 50 to 60 grams and its taste and smell are disgusting and therefore, its oral administration is difficult in many cases. Since shark cartilage is a mixture of mucopolysaccharides, there are no other routes of administration other than the oral route. What is needed is that, during oral administration, under strong acid conditions, shark cartilage remains insoluble in the stomach. , But in the absorption position, it is dissolved in ______3___ This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm)-(Please read the precautions on the back before filling this page)
IT 經濟部中央標準局員工消費合作社印製 577745 A7 B7 五、發明説明（2 ) 腸內。對癌症病人而言其已經變得非常虛弱，口服施藥常 常很困難去進行。在沒有解決如味道、氣味及劑量問題 前，鯊魚軟骨不能夠被應用作爲一種有效的抗癌藥劑而被 廣泛地使用。 由上所述，一種使鯊魚軟骨可供實務使用作爲一種有 效的腫瘤血管生成抑制劑之技術已經被強烈地需要。 另一方面，白血球間質1 2 ( I L — 1 2)被發現爲 一種具有活化NK (天然殺傷）細胞效果之細胞素。隨後 之硏究顯示其可刺激生長及活化具有專一性細胞毒素活性 之T細胞（殺傷性T細胞）以對抗腫瘤細胞且，此外，其 促進可活化殺傷性τ細胞之干擾素r ( I FN— r)之產 生。因此，其可作爲一種有效用來治療癌症病人之物質已 引起很多注意。 最近，在美國，藉由使用基因遺傳工程技術（重組白 血球間質1 2 ( Γ t — I L 一 1 2 ))已使得大量生產白 血球間質1 2成爲可能。藉由使用基因轉移技術已嘗試將 此白血球間質1 2之基因密碼直接導入癌細胞，因而白血 球間質1 2可被直接地施於癌細胞。 當藉由證實白血球間質12的使用其包括自活體生物 的外部施與白血球間質12可用來抑制腫瘤生長或致使其 消失的一種方法時，另一種方法爲包括致使生物體誘導自 體白血球間質1 2。此自體白血球間質1 2其優勢在於其 不會招致引起異常免疫反應的危險。此外，其具有高敏感 度。因此，其被期望能產生一種顯著的腫瘤縮小或根除的 _____4__ 本紙張尺度適用中國國家標準（CNS ) A4規格（210x 297公釐1 ： 一 -- (請先閲讀背面之注意事項再填寫本頁) 計 辞 577745 A7 B7 五、發明説明（3 ) 效果。 發明摘述 (請先閱讀背面之注意事項再填寫本頁) 參考前述之先前技藝，本發明的一個目的爲提供能夠 施與鯊魚軟骨之藥物組成物，其爲一種腫瘤血管生成抑制 劑，可作爲一種抗癌藥劑。本發明的另一目的爲提供一種 非常有用的抗癌組成物，其中腫瘤血管生成抑制劑和白血 球間質1 2誘導劑合倂使用。 本發明關於一種腫瘤血管生成抑制劑其包括包埋於脂 肪或油基質之精細分割鯊魚軟骨，該基質的表面被進一步 以另一種與脂肪或油基質不同熔點之脂質所包覆。 發明詳述 以下爲本發明詳細說明。 經濟部中央標準局員工消費合作社印製 鯊魚軟骨，其爲本發明之腫瘤血管生成抑制劑的主要 成分，必須是精細分割的形式。在本說明書中，該“精細 分割的形式”之名詞其意思爲該粒徑爲1至9 0微米。爲 製備如此精細分割之鯊魚軟骨，其須要精細地細分商業等 級之鯊魚軟骨。其精細細分必須在低溫進行乃因爲當於超 過6 0°C之溫度加熱時，包含在鯊魚軟骨中作爲有效成分 之蛋白質及其他活性成分會熱變性且失去它們的效果。該 低溫精細細分之進行可藉由，例如，使鯊魚軟骨接受一 1 9 6 °C之液態氮瞬間冷凍，以截切器型式粉碎機如定位礙 磨機（O r i e n t m i 11 )約略地細分冷凍塊，然後在冷凍的狀 態中以高速旋轉型式粉碎機如.葉輪碾磨機（ImPellef Mill)細分該約略細分塊及進一步以空氣一噴射型式粉碎 _ 5 _ 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） 經濟部中央標準局員工消費合作社印製 577745 A7 B7 五、發明説明（f) 機如噴射碾磨機（j e t m i 11 )細分該結果塊。 在製備本發明之腫瘤血管生成抑制劑中，該前述之精 細分割鯊魚軟骨爲包埋於一脂肪或油基質中。 構成該脂肪或油基質之脂肪或油並未限定在任何所提 供之特別種類，其爲一般使用作爲藥物目的之一種。因 此，其可適當地選擇，例如，自硬蔬菜油如大豆油、菜 油、棕櫚油、玉蜀黍油及棉子油；硬動物油如牛脂、豬油 及魚油；及其混合物中。在它們之中，具有熔點4 2 °C至 5 6 °C之脂肪及油類爲較佳。如果需要，一種抗氧化劑如 維生素E或兒茶素（catechin)可合倂使用。 用來於作爲基質之脂肪或油內包埋該精細分割鯊魚軟 骨的方法並未特別限定。例如，該包埋係藉由使在液化狀 態之脂肪或油之攪動混合物及精細分割之鯊魚軟骨在高速 攪動型式製粒機內製成粒狀而完成。 以上述方法產生之包含包埋於其中之精細分割鯊魚軟 骨的脂肪或油基質然後進一步接受以另一種與該脂肪或油 不同熔點之脂質包覆表面。該使用於本發明實施之脂質可 選擇自那些與進行步驟中使闱於製備脂肪或油基質之脂肪 或油不同熔點之脂質中，且普遍地那些脂質具有較高熔 點。具體實施例中，所描述的可爲蠟如巴西棕櫚蠟、米蠟 及蜂蠟；硬動物或植物油；脂肪酸如硬脂酸及棕櫚酸；天 然樹脂如蟲膠片；及長鏈醇類如棕櫚醇及硬脂醇。在進行 包覆時，例如，粉末塗敷技術，可被使周。 另一種方法可被使用於製備本發明之腫瘤血管生成抑 6 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） «I--·----ΦII C请先閱讀背面之注意事項再填寫本頁) 訂 #1 577745 Α7 Β7 五、發明説明（c) 制劑包括’例如，將該精細分割鯊魚軟骨分散在事先熔解 之脂肪或油內，使膠體溶液受噴霧冷卻及然後包覆該產生 之粉末，其係在一滾動製粒機內，以另一種不同於該脂肪 或油之熔點的脂質進行。 因此製備之本發明的腫瘤血管生成抑制劑其包含鯊魚 軟骨作爲主要的活性成分且其味道及氣味完全地被遮蔽， 藉此使口服施藥的困難能被完全地淸除。此外，製備之腫 瘤血管生成抑制劑顯示出減低的表面摩擦且，因此，其具 有改善的流動性及減低的粘連性質，因而其非常適合口服 施藥。此外，其能被設計成在胃內之強酸條件下不溶及在 腸內其吸收位置可溶。 在被使用於本發明實行之精細分割鯊魚軟骨中，粒子 具有粒徑不大於3 2微米佔至少9 9 · 5重量％。在此處後 之實施例中可作詳細解釋，該精細分割鯊魚軟骨之粒徑對 本發明的效果具有很大的影響。當粒徑太大時，該腫瘤血 管生成抑制活性降低。本發明提到之一種用來使鯊魚軟骨 粒徑一致的方法，例如，爲一種方法其包括將精細分割鯊 魚軟骨通過一具有所欲篩孔之篩子。 經濟部中央標準局員工消費合作社印製 (請先閱讀背面之注意事項再填寫本頁) 雖然被使用在本發明之實施中作爲鯊魚軟骨原料之鯊 魚並未被限制在任何特定種類，但獲得自藍黨（大靑鯊 (P r i ο n a c e g 1 a u c a ))之繁軟骨爲最佳。藍黨之軟骨包 含大量活性成分蛋白質及共多糖ly s a c c h a r i d e ) 且特別地適合使用。美國產爲產生自許多種類 之鯊魚的混合物，因此其顯示偏差。另一缺點爲它 ' 、...../---------… _7__________ 本紙張尺度適用中國國家標準（ CNS ) Λ4規格（210Χ 297公釐1 ^ ~ 577745 Λ7 B7 五、發明説明（心） 的效果相當低。 本發明已建立出本發明之腫瘤血管生成抑制劑不只具 有腫瘤血管生成抑制活性但亦有可靠的止痛活·注。患有癌 症之病患，特別是末期癌症，其普遍垲抱怨劇痛。因此， 當本發明之腫瘤血管生成抑制劑被作爲抗癌藥劑而施與於 抱怨劇痛的癌症病患時，其抗癌活性及止痛活性可產生出' 一種協乘效果，藉此可獲得非常有效的治療效杲。 本發明之腫瘤血管生成抑制劑被硏究其如何產生腫瘤 血管生成抑制活性及，因此，如此處後於實施例中之詳細 解釋，其暗示包含脫作用（apoptosis)。 當本發明之腫瘤血管生成抑制劑作爲一種抗癌藥劑非 常有效時，其同樣地具有止痛效果，能被使用在其他藥物 用途之領域。因此，在第二個觀點內，本發明提供一種藥 物組成物其包括本發明之腫瘤ί&管生成抑制劑。 此外，一種用來治療癌症的方法，其包括施與一足以 抑制腫瘤血管生成之劑量的本發明之腫瘤血管生成抑制 劑，該方法亦落於本發明之範疇內。 經濟部中央標準局員工消費合作社印製 (請先閱讀背面之注意事項再填寫本頁) 本發明之藥物組成物其特徵在於其包含腫瘤血管生成 抑制劑及白ιΐα球間質1 2誘導劑。該藥物組成物反映本發 明之第二觀點。以下詳述本發明之第二觀點。 在說明書中，該“白血球間質1 2誘導劑”之名詞包 括，在其意思內，不只是指能誘導在活體內之白血球間質 1 2 ( I L — 1 2 )之物質但苏包括白血球間質1 2本 身，白血球間質1 2基因引入癌細胞，r t 一 I L — 1 2 ______8 _ 本紙張尺度適用中國國家標準（CNS ) A4規格（210X 297公釐） 經濟部中央標準局員工消費合作社印製 577745 Λ 7 Β7 '——— --------------— -—-- 五、發明説明（Ί ) 及，總而言之，所有能在活體內誘導產生白血球間質1 2 之物質，並沒有任何特別限制。 如此之實施例，本發明提到活性半纖維素化合物（Α H C C s )及其類似物。該活性半纖維素化合物早已知其 可獲得自經由酵素處理包含在蕈類菌絲細胞壁之植物纖維 而作爲生理活性物質及包含雜葡聚糖，例如’沒一（1 — 3) D—葡聚糖，彡一（1 — 6) D —葡聚糖及α — (1 —4 ) D —葡聚糠；葡聚糖肽，蛋白葡聚糖，外源凝集 素，核酸，不消化的多醣等等。 當白血球間質1 2誘導劑被使用於本發明內，除了上 述活性半纖維素化合物外，其可提到蕈類菌絲成分。該覃 類菌絲成分並未限定於任何特別種類但包括，其中，P S Κ，其爲變色多孔菌（Polyporus versicolor)之菌絲 成分可使用作爲抗癌藥劑，S P G ，其爲裂褶菌 (Schizophyllum commune )之 菌絲成分，及香菇多 糖，其爲香薛（Lentinula edodes)之菌絲成分。 作爲如此之蕈類菌絲成分，其可進一步提到松蕈 (agarics ) ，靈芝（Ganoderma lucidum ) ，Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of IT and Economy 577745 A7 B7 V. Description of the invention (2) Intestine. It has become very weak for cancer patients, and oral administration is often difficult. Without addressing issues such as taste, odor, and dosage, shark cartilage could not be widely used as an effective anticancer agent. From the above, a technique for making shark cartilage available for practical use as an effective tumor angiogenesis inhibitor has been strongly demanded. On the other hand, Leukocyte Interstitial 1 2 (IL-12) was found to be a cytokine with the effect of activating NK (natural killer) cells. Subsequent research has shown that it can stimulate growth and activate T cells (killer T cells) with specific cytotoxic activity to fight tumor cells and, in addition, it promotes interferon r (I FN— r). Therefore, it has attracted much attention as a substance effective for treating cancer patients. Recently, in the United States, the use of genetic engineering technology (recombinant white blood cell interstitial 12 (Γ t-I L-1 2)) has made it possible to mass produce white blood cell interstitial 12. Attempts have been made to directly introduce the genetic code of this white blood cell stroma 12 into cancer cells by using gene transfer technology, so white blood cell stroma 12 can be directly applied to cancer cells. When the use of leukocyte mesenchyme 12 is demonstrated to include the use of an external application of leukocyte mesenchyme 12 from a living organism to inhibit tumor growth or cause it to disappear, another method includes causing the organism to induce the autologous leukocyte Quality 1 2. This autologous leukocyte mesenchyme 12 has the advantage that it does not incur the risk of causing an abnormal immune response. In addition, it is highly sensitive. Therefore, it is expected to produce a significant tumor shrinkage or eradication. _____4__ This paper size applies the Chinese National Standard (CNS) A4 specification (210x 297 mm1: one-(Please read the precautions on the back before filling in this (Page) Words 577745 A7 B7 V. Description of the invention (3) Effect of the invention (please read the notes on the back before filling this page) With reference to the previous art, one object of the present invention is to provide shark cartilage The pharmaceutical composition is a tumor angiogenesis inhibitor and can be used as an anticancer agent. Another object of the present invention is to provide a very useful anticancer composition, wherein the tumor angiogenesis inhibitor and the white blood cell interstitial 1 2 An inducer is used in combination. The present invention relates to a tumor angiogenesis inhibitor comprising finely divided shark cartilage embedded in a fat or oil matrix, the surface of which is further coated with another lipid having a melting point different from that of the fat or oil matrix Detailed description of the invention The following is a detailed description of the invention. Shark cartilage is printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs, The main component of the tumor angiogenesis inhibitor of the present invention must be in a finely divided form. In this specification, the term "finely divided form" means that the particle size is 1 to 90 micrometers. In order to prepare this Finely divided shark cartilage needs to finely subdivide commercial grade shark cartilage. Its fine subdivision must be performed at low temperature because when heated at a temperature exceeding 60 ° C, proteins and other ingredients contained in shark cartilage as an active ingredient and others The active ingredients will be thermally denatured and lose their effect. The low temperature fine subdivision can be performed, for example, by subjecting shark cartilage to a liquid nitrogen of 196 ° C for instant freezing, and using a cutter-type grinder such as positioning to prevent grinding Machine (Orientmi 11) to roughly divide the frozen block, and then in a frozen state, use a high-speed rotating type pulverizer such as an impeller mill (ImPellef Mill) to divide the roughly divided block and further pulverize it with an air-jet type _ 5 _ This paper size applies to China National Standard (CNS) A4 (210X297 mm) Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 577745 A7 B7 V. Description of the invention (f) A machine such as a jet mill 11 (jetmi 11) subdivides the result block. In preparing the tumor angiogenesis inhibitor of the present invention, the aforementioned finely divided shark cartilage is embedded in a fat Or oil base. The fat or oil constituting the fat or oil base is not limited to any particular kind provided, and it is one that is generally used for pharmaceutical purposes. Therefore, it can be appropriately selected, for example, a self-hardening vegetable oil Such as soybean oil, vegetable oil, palm oil, corn oil, and cottonseed oil; hard animal oils such as tallow, lard, and fish oil; and mixtures thereof. Among them, fats having a melting point of 4 2 ° C to 56 ° C and Oils are preferred. If desired, an antioxidant such as vitamin E or catechin can be used in combination. A method for embedding the finely divided shark cartilage in a fat or oil as a matrix is not particularly limited. For example, the embedding is accomplished by making the agitation mixture of fat or oil in a liquefied state and finely divided shark cartilage into granules in a high-speed agitation type granulator. The fat or oil matrix containing finely divided shark cartilage embedded in the above-mentioned method is then further subjected to coating the surface with another lipid having a melting point different from that of the fat or oil. The lipids used in the practice of the present invention may be selected from lipids having a melting point different from that of fats or oils that are used in the preparation of fats or oil bases, and generally those lipids have higher melting points. In specific embodiments, the descriptions may be waxes such as carnauba wax, rice wax, and beeswax; hard animal or vegetable oils; fatty acids such as stearic acid and palmitic acid; natural resins such as shellac; and long-chain alcohols such as palmitol and Stearyl alcohol. When coating, for example, powder coating techniques can be applied. Another method can be used to prepare the tumor angiogenesis inhibitor of the present invention. 6 The paper size is applicable to China National Standard (CNS) A4 (210X297 mm). «I-- · ---- ΦII C Please read the note on the back first Please fill in this page again) Order # 1 577745 Α7 Β7 V. Description of the invention (c) The preparation includes' for example, dispersing the finely divided shark cartilage in previously melted fat or oil, subjecting the colloidal solution to spray cooling and then coating The resulting powder is processed in a rolling granulator with another lipid that has a melting point different from the fat or oil. Therefore, the tumor angiogenesis inhibitor of the present invention is prepared which contains shark cartilage as a main active ingredient and its taste and odor are completely masked, so that the difficulty of oral administration can be completely eliminated. In addition, the prepared tumor angiogenesis inhibitor exhibits reduced surface friction and, therefore, it has improved fluidity and reduced adhesion properties, making it very suitable for oral administration. In addition, it can be designed to be insoluble under strong acid conditions in the stomach and soluble in its absorption site in the intestine. In the finely divided shark cartilage used in the practice of the present invention, the particles have a particle size of not more than 32 microns and occupy at least 99.5% by weight. As will be explained in detail in the following examples, the particle size of the finely divided shark cartilage has a great influence on the effect of the present invention. When the particle size is too large, the tumor angiogenesis inhibitory activity decreases. The invention refers to a method for making shark cartilage uniform in particle size, for example, a method which includes passing finely divided shark cartilage through a sieve having a desired sieve opening. Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the notes on the back before filling this page) Although the sharks used as raw materials for shark cartilage in the implementation of the present invention are not restricted to any specific species, they are obtained from The blue carcass (P ri ο naceg 1 auca) has the best cartilage. The cartilage of the blue party contains a large amount of active ingredient proteins and copolysaccharides (ly s a c c h a r i d e) and is particularly suitable for use. Produced in the United States is a mixture produced from many types of sharks, so it shows deviations. Another disadvantage is that it ', ..... / ---------... _7__________ This paper size applies the Chinese National Standard (CNS) Λ4 specification (210 × 297 mm 1 ^ ~ 577745 Λ7 B7 V. Invention The effect of (heart) is quite low. The present invention has established that the tumor angiogenesis inhibitor of the present invention not only has tumor angiogenesis inhibitory activity, but also has a reliable analgesic activity · injection. Patients with cancer, especially terminal cancer It generally complains of severe pain. Therefore, when the tumor angiogenesis inhibitor of the present invention is administered as an anticancer agent to cancer patients complaining of severe pain, its anticancer activity and analgesic activity can produce a kind of synergy This effect can be used to obtain a very effective therapeutic effect. The tumor angiogenesis inhibitor of the present invention has been investigated how to produce tumor angiogenesis inhibitory activity, and therefore, as explained in detail in the examples hereafter, it implies that Including Apoptosis. When the tumor angiogenesis inhibitor of the present invention is very effective as an anticancer agent, it also has an analgesic effect and can be used in other pharmaceutical applications. Therefore, in a second aspect, the present invention provides a pharmaceutical composition comprising the tumor angiogenesis inhibitor of the present invention. In addition, a method for treating cancer, which comprises administering a tumor vessel sufficient to inhibit tumor blood vessels. The generated dose of the tumor angiogenesis inhibitor of the present invention is also within the scope of the present invention. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) The pharmaceutical composition is characterized in that it contains a tumor angiogenesis inhibitor and an inducer of white interstitial stromal 12. The pharmaceutical composition reflects the second aspect of the present invention. The second aspect of the present invention is described in detail below. In the specification, The term "leukocyte interstitial 12 inducer" includes, within its meaning, not only substances capable of inducing leukocyte interstitial 1 2 (IL-12) in vivo, but Su includes leukocyte interstitial 1 2 itself , Leukocyte Interstitial 1 2 Genes Introduced into Cancer Cells, rt-IL — 1 2 ______8 _ This paper size applies to China National Standard (CNS) A4 (210X 297 mm) Economy Printed by the Consumer Standards Cooperative of the Ministry of Standards of the People's Republic of China 577745 Λ 7 Β7 '———— --------------— -——- V. Description of the Invention (Ί) And, all in all, all The substance that induces the production of leukocyte mesenchyme 12 in vivo is not particularly limited. In this embodiment, the present invention refers to active hemicellulose compounds (A HCC s) and analogs thereof. The active hemicellulose compounds have been known for a long time. It can be obtained from the treatment of plant fibers contained in the cell wall of mushroom mycelium by enzymes as a physiologically active substance and contains heteroglucan, such as' Michi (1-3) D-glucan, Zanyi (1-6 ) D-glucan and α- (1-4) D-glucan bran; glucan peptide, protein glucan, exogenous lectin, nucleic acid, non-digestible polysaccharide and so on. When the leukocyte interstitial 12 inducer is used in the present invention, in addition to the above-mentioned active hemicellulose compound, it may mention a mushroom mycelium component. The mycelium mycelium component is not limited to any particular species but includes, among them, PS K, which is a hyphae component of Polyporus versicolor, can be used as an anticancer agent, SPG, which is Schizophyllum commune), and shiitake mushroom polysaccharide, which is the mycelium component of Lentinula edodes. As such a mushroom mycelium component, it may further mention agarics, Ganoderma lucidum,
Alb atr e llu s confluens，洋薛(Agaricus b 1 a z e i )， 巨大口磨（Tricholoma giganteum ) ’ 纖扎囷 (Inonotus obliquus ) ，奇果菌（Grifola frondosa)，猴頭蘇（Hericium erinaceum)，鮑魚 薛（Pleurotus ostreatus )，靈芝（Ganoderma lucidum)，扇薛（Panellus serotinus) ’ 層孔菌 -______9__ 本紙張尺度適用中國國家標準（CNS ) A4規格（210X Μ?公釐） --.--·-----Φ-- (請先閲讀背面之注意事項再填寫本頁) 訂 577745 A7 __—_BT___ 五、發明説明（2 ) (請先閱讀背面之注意事項再填寫本頁) (Phellinus )，樺革樞菌（Lenzites betulinus )，松 口磨(Tricholoma matsutake )，多孔菌 (Perenniporia fraxinea ) ，F 1 a m m u 1 i n a v e i u t i p e s 等等之菌絲成分。 被使用於本發明之白血球間質1 2誘導劑其進一步包 括釀騰鍵球菌（Streptococcus pyogenes)或溶血性鍵 球菌（Streptococcus haemolitycus)之細胞成分。， 其提到已知抗癌藥劑，例如，0 K — 4 3 2 (p i c i b a n i 1 )，作爲細胞成分。 有物質已被知可作爲生物反應調節物（BRMs )。 上述之白血球間質1 2誘導劑具有殺傷性T細胞活化促進 效果。因此，在活體內產生之殺傷性T細胞藉由該白血球 間質1 2誘導劑而被活化。 經濟部中央標準局員工消費合作社印製 另一方面，前面詳述之腫瘤血管生成抑制劑抑制由癌 細胞（腫瘤細胞）導致之新血管增生。當新血管增生被抑 制時，該腫瘤細胞，在血液供應抑制之壓力下，表現於其 表面之黏連因子如Fa s抗原、CD80及CD86。殺 傷性T細胞能識別如F a s抗原、C D 8 0、C D 8 6及 其他作爲抗原之黏連因子。因此，可藉由殺傷性τ細胞容 易地識別出那些在一狀態內之表現黏連因子，且腫瘤細胞 因此修飾及現在可容易地識別並容易地被殺傷性Τ細胞攻 擊。 若存在於如此之狀況，上述之白血球間質1 2與只使 甩白血球間質1 2誘導劑之病例相比能更顯著地攻擊腫瘤 本紙張尺度適用中國國家標準（CNS ) Α4規格（210Χ 297公釐）、 _10____ 577745 __B7__ 五、發明説明（7 ) 細胞。本發明之腫瘤血管生成抑制劑及白血球間質1 2誘 導劑之協乘作用爲基於上述之機制且運用強抗癌效果’其 能導致脫噬作用而使腫瘤細胞被破壞。此已被本案發明人 首先發現。 本發明之藥物組成物之劑量形式爲施與人類或其他動 物，其並未限定在任何特別之一種形式’但可是任何一種 普遍周於不同的藥物及被載劑所支持之劑量形式。 作爲此種載劑，至少一種之固體，半固體，液體稀釋 劑，塡充劑及其他配方輔助物被以比例使用，例如，0 . 1 %至99. 5%，較佳地爲，0.5%至90%。本發阴之 藥物組成物可被完全地以口服或非口服使用。其所提到， 作爲非口服途徑，例如，局部（如組織內），皮下，肌 內，動脈內或靜脈內，及直腸施藥。該適合作爲此施藥途 徑之劑量形式可經由傳統技術裝置製備。 經濟部中央標率局員工消費合作社印製 (請先閱讀背面之注意事項再填寫本頁) 當該組成物被使用作爲抗癌藥劑時，例如，其所欲決 定之劑量經由考慮病患之年齡及體重，施藥途徑，疾病， 其嚴重及其他因子而決定。一般地，然而，對成入之口服 劑量’以活性物質之方式表示爲在1至1 〇克/天之範圍 內，較佳地爲’ 5至6克/天。該非口服劑量，其根據施 藥途徑而可能大大地不同，一般可在1 0 0至1 0，Q 0 0毫克/天之範圍內，較佳地爲，2至5克/天。在一些 例子中、較小之劑量可能足夠而，相反地，在特定例子內 可能需要較大劑量。每天劑量可以二或四分次劑量施與。 該口服施藥可被瞭解爲使用固體或液體單位劑量形 ___11__ 尺度適用中國國家標準（CNS ) Λ4規格（210X 297公趁）、 一 經濟部中央標準局員工消費合作社印製 577745 A7 B7 五、發明説明（π ) 式，例如，以大體積粉末，粉末，顆粒，錠劑，膠囊，糖 漿，酏劑，懸浮液等等之形式。 該大體積粉末可經由分割活性物質至一合適精細而製 造。該粉末可經由分割活性物質至一合適精細及混合以同 樣方式精細分割之藥物載劑，例如，一種可食的碳水化合 物如澱粉或甘露醇或一些其他合適之賦形劑而製造。若必 要，可加入其他的添加物如調味劑，保存劑，分散劑，著 色劑，香料及其類似物。 該膠囊可經由製備來自上述方式所獲得之大體積粉末 或粉末之細粒而製造，或錠劑及包裝該產生之細粒於膠囊 殼內，如明膠膠囊殼。在塡充之前，一種潤滑藥及/或流 動性改進劑，例如，膠質二氧化矽，滑石，硬脂酸鎂，硬 脂酸鈣或固體聚乙二醇，可選擇性地加入。該藥物功效， 當使用膠囊時，可經由加入崩解劑或增溶劑而改善，其例 如，羧甲基纖維素，羧甲基纖維素鈣，低取代羥基丙基纖 雜素，croscarmellose sodium，竣基澱粉鈉，碳酸 鈣，碳酸鈉或其類似物。 該軟膠囊可經由於植物油，聚乙二醇，甘淄或表面活 性劑內懸浮及分散該精細分割之活性物質及包裝該產生之 膠溶液於明膠薄片內而製備。 該細粒可經由混合粉末形式之活性物質及賦形劑及上 述所提之崩解劑，加入濕潤劑（例如，糖漿，澱粉糊劑， 阿拉伯樹膠，纖維素溶液，聚合物溶液），若必要可一起 加入結合劑（例如，羧甲基纖維素鈉，羥基丙基纖維素， ______12_ 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） -----.---------.--了______I (請先閱讀背面之注意事項再填寫本頁) 577745 Λ7 B7 __ —— ' _______ 五、發明説明（I丨） (請先閱讀背面之注意事項再填寫本頁) 甲基纖維素，羥基丙基S基纖維素，明膠，聚乙烯比咯烷 酮，聚乙烯醇），揉捏該產生之混合物及強迫地將該混合 物通過一篩子而製備。除了此粉末製粒法外，細粒亦可經 由初步將該混合物送至打錠機及磨碎該產生之不完全形式 之渣而製造。可預先加入溶解延緩劑（例如，石蠟，蠟， 硬化蓖麻油），再吸收劑（例如，四級鹽）或吸附劑（例 如，自土，高嶺土，磷酸二鈣）。 該錠劑可經由加入硬脂酸，硬脂酸鹽，滑石，礦油或 其類似潤滑劑至以上述方法獲得之細粒及然後將該產生之 混合物打錠而製備。因而製造之素錠可被提供膜包衣或糖 衣。 經濟部中央標準局員工消費合作社印製 錠劑製造亦可經由混合本發明之活性物質及可流動情 性載體，直接打錠，而沒有如上述之顆粒化或渣形成之步 驟。亦可使用由密閉蟲膠片製成之透明或半透明防護包 衣，由糠或聚合物製成之包衣，及由蠟製成之磨光包衣。 其他口服劑量形式，例如，糖漿，藥液酏及懸浮液，亦可 被製成單位劑量形式如一定量藥物其包含一定量之活性物 質。該糖漿經由溶解活性物質於適當調味之水性溶液而被 製造。該藥液酏經由使用非毒性醇類載體而被製造。該懸 浮液經由分散活性物質於毒性載體而製成配方。可同樣地 任意加入懸浮劑或乳化劑（例如，乙氧基異硬脂基醇，多 氧乙烯山梨糖醇酯），保存劑，調味劑（例如，薄荷油， 糖精）及/或其類似物。 如必須，該用來口服施藥之單位劑量配方可被微包 ___13_____ 本紙張尺度適用中國國家標準（CNS ) A4規格（210X 297公趁) " — 57774sAlb atr e llu s confluens, Agaricus b 1 azei, Tricholoma giganteum 'Inonotus obliquus, Grifola frondosa, Hericium erinaceum, Abalone Xue (Pleurotus ostreatus), Ganoderma lucidum, Panellus serotinus' Shell-fungus -______ 9__ This paper standard applies to China National Standard (CNS) A4 specification (210X Μ? Mm) -------- --- Φ-- (Please read the notes on the back before filling this page) Order 577745 A7 __—_ BT___ V. Description of the invention (2) (Please read the notes on the back before filling this page) (Phellinus), birch Mycelium components of Lenzites betulinus, Tricholoma matsutake, Perenniporia fraxinea, F 1 ammu 1 inaveiutipes, etc. The leukocyte interstitial 12 inducer used in the present invention further includes cell components of Streptococcus pyogenes or Streptococcus haemolitycus. It mentions known anticancer agents, such as 0K-4 3 2 (p i c i b a n i 1), as cellular components. Some substances have been known as biological response modifiers (BRMs). The aforementioned leukocyte interstitial 12 inducer has a killer T cell activation promoting effect. Therefore, the killer T cells produced in vivo are activated by the leukocyte interstitial 12 inducer. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs On the other hand, the tumor angiogenesis inhibitors detailed above inhibit the proliferation of new blood vessels caused by cancer cells (tumor cells). When neovascular hyperplasia is suppressed, the tumor cells, under the pressure of blood supply inhibition, show adhesion factors such as Fas antigen, CD80 and CD86 on their surface. Killer T cells recognize antigens such as Fas antigen, CD80, CD86 and other antigens. Therefore, killer τ cells can easily recognize those expressing adhesion factors in a state, and tumor cells are thus modified and can now be easily identified and easily attacked by killer T cells. If existing in such a condition, the above-mentioned leukocyte interstitial 12 can attack the tumor more significantly than the case where only the leukocyte interstitial 12 inducer is attacked. This paper applies the Chinese National Standard (CNS) A4 specification (210 × 297). Mm), _10____ 577745 __B7__ 5. Description of the invention (7) Cell. The synergistic effect of the tumor angiogenesis inhibitor and the leukocyte interstitial 12 inducer of the present invention is based on the above-mentioned mechanism and uses a strong anti-cancer effect ', which can lead to a dephagizing effect and destroy tumor cells. This has been first discovered by the inventor of this case. The dosage form of the pharmaceutical composition of the present invention is for administration to humans or other animals, and it is not limited to any particular form 'but may be any dosage form generally supported by different drugs and carriers. As such a carrier, at least one kind of solid, semi-solid, liquid diluent, tincture and other formulating aids are used in proportion, for example, 0.1% to 99.5%, preferably 0.5%. To 90%. The pharmaceutical composition of the present Yin can be used completely orally or parenterally. It is mentioned as a non-oral route, for example, topical (such as intra-tissue), subcutaneous, intramuscular, intra-arterial or intravenous, and rectal administration. The dosage form suitable for this route of administration can be prepared by means of conventional technology. Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the notes on the back before filling this page) And weight, application route, disease, its severity and other factors. Generally, however, the oral dose for ingestion is expressed in the form of an active substance in the range of 1 to 10 g / day, preferably '5 to 6 g / day. The non-oral dosage may vary greatly depending on the route of administration, and may generally be in the range of 100 to 100 mg / day, preferably 2 to 5 g / day. In some cases, smaller doses may be sufficient, and conversely, larger doses may be required in specific cases. The daily dose can be administered in two or four divided doses. The oral administration can be understood as the use of solid or liquid unit dosage forms ___11__ scales applicable to Chinese National Standards (CNS) Λ4 specifications (210X 297) while being printed by a Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 577745 A7 B7 Description of the invention The formula (π) is, for example, in the form of a large volume of powder, powder, granules, lozenges, capsules, syrups, elixirs, suspensions and the like. The bulk powder can be made by dividing the active substance to a suitable fineness. The powder can be made by dividing the active substance into a suitable finely divided and mixed pharmaceutical carrier which is finely divided in the same manner, for example, an edible carbohydrate such as starch or mannitol or some other suitable excipient. If necessary, other additives such as a flavoring agent, a preserving agent, a dispersing agent, a coloring agent, a fragrance, and the like may be added. The capsules can be manufactured by preparing large-volume powders or fine particles obtained from the above methods, or tablets and packaging the resulting fine particles in a capsule shell, such as a gelatin capsule shell. Prior to filling, a lubricant and / or flow improver, such as colloidal silica, talc, magnesium stearate, calcium stearate, or solid polyethylene glycol, may be optionally added. The efficacy of the drug can be improved when capsules are used by adding disintegrants or solubilizers, such as carboxymethyl cellulose, carboxymethyl cellulose calcium, low-substituted hydroxypropylcellulose, crocarmellose sodium, end Sodium starch, calcium carbonate, sodium carbonate or the like. The soft capsule can be prepared by suspending and dispersing the finely divided active substance in vegetable oil, polyethylene glycol, ganzi or surfactant, and packaging the resulting gel solution in a gelatin sheet. The fine particles can be mixed with the active substance and excipients in powder form and the disintegrating agents mentioned above, and added with a wetting agent (for example, syrup, starch paste, gum arabic, cellulose solution, polymer solution), if necessary Binders can be added together (for example, sodium carboxymethyl cellulose, hydroxypropyl cellulose, ______12_ This paper size applies to China National Standard (CNS) A4 specifications (210X297 mm) -----.----- ----.-- ______I (Please read the notes on the back before filling this page) 577745 Λ7 B7 __ —— '_______ V. Description of the Invention (I 丨) (Please read the notes on the back before filling in this Page) Methyl cellulose, hydroxypropyl S-based cellulose, gelatin, polyvinylpyrrolidone, polyvinyl alcohol), prepared by kneading the resulting mixture and forcibly passing the mixture through a sieve. In addition to this powder granulation method, fine granules can also be produced by initially feeding the mixture to a beater and grinding the resulting incomplete form of slag. Dissolution retarders (e.g., paraffin, wax, hardened castor oil), reabsorbents (e.g., quaternary salts) or adsorbents (e.g., clay, kaolin, dicalcium phosphate) can be added beforehand. The lozenges can be prepared by adding stearic acid, stearates, talc, mineral oil or similar lubricants to the fine particles obtained in the above method, and then tableting the resulting mixture. The manufactured tablets can thus be provided with a film coating or a sugar coating. The manufacturing of tablets by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs can also be performed directly by mixing the active substance of the present invention and the flowable emotional carrier, without the steps of granulation or slag formation as described above. Transparent or translucent protective coatings made of closed shellac, coatings made of bran or polymer, and polished coatings made of wax can also be used. Other oral dosage forms, such as syrups, medicinal solutions, and suspensions, can also be made into unit dosage forms such as a certain amount of a drug which contains a certain amount of active substance. This syrup is produced by dissolving an active substance in a suitably flavored aqueous solution. This medicinal liquid tincture is manufactured by using a non-toxic alcohol carrier. This suspension is formulated by dispersing the active substance in a toxic carrier. Suspensions or emulsifiers (for example, ethoxyisostearyl alcohol, polyoxyethylene sorbitol esters), preservatives, flavoring agents (for example, peppermint oil, saccharin) and / or the like can be similarly added arbitrarily. . If necessary, the unit dose formula used for oral administration can be micro-packaged. ___13_____ This paper size applies to China National Standard (CNS) A4 specification (210X 297). &Quot; — 57774s
五、發明説明（A ) 膜。配方藉由進一歩包衣或將活性物質嵌入聚合物或蠟， 其作用的期間可被延長且可達到持續釋放。 該皮下，肌內，動脈內或靜脈內施藥可經由製造液態 單位劑量形式，例如，溶液或懸浮液之可注射組成物而進 行。其可經由溶解或懸浮一定量之活性物質於適合注射目 的之非毒性液體載體，例如，水性或油脂性溶劑及滅菌該 產生之溶液或懸浮液而被製造。其亦可分配一定量之活性 物質以粉末或冷凍乾燥形式進入小瓶及然後使該小瓶及其 內容物滅菌及密閉。在此例子中，在施藥前，溶解或混合 儲備小瓶及載體。可加入非毒性鹽類或鹽類溶液使該可注 射組成物等張。此外，可附加使周穩定劑，保存劑，懸浮 劑，乳化劑及/或其類似物。 該直腸劑量形式可經由混合活性物質與疏水性或親水 性坐藥基劑，例如，聚乙二醇，可可脂，高級酯類（例 如，棕櫚酸肉豆蔻酯）或其混合物及揉捏該產生之混合物 而被製備。 如上述，本發明之腫瘤血管生成抑制劑無鯊魚軟骨之 使人厭惡之味道及氣味，可抗酸及只在腸內溶解，且可方 便地經由口服途徑施藥。此外，其具有可靠的抗癌活性及 止痛活性及作爲藥物組成物非常有效。 本發明之腫瘤血管生成抑制劑及白血球間質一 1 2誘 導物合倂使用顯示出非常高的抗癌活性且因此製備成藥物 組成物非常有效。 本發明進行最佳模式 _u___ 本紙張尺度適用中國國家標準（CNS ) A4規格（210X 297公釐） " ~ ϋϋ r I - : I - I 1 二 ϋ (請先閱讀背面之注意事項再填寫本頁) 订 經濟部中央標準局員工消費合作社印製 577745 B7 _ _ 五、發明説明（Ο ) 下列生產實施例及實施例作進一步詳細列舉說明本發 明。它們，然而，絕不因而限制本發明的範圍。 生產實施例1 包埋鯊魚軟骨之油或脂肪基質之包覆配方的製造 將平均粒徑2 7微米之6 0重量份之精細分割鯊魚軟 骨裝入高速攪袢製粒機（高速混合器，深江工業社製，） 及，保持製粒機之機內溫度爲1 〇°C及在2 0至1 0 0 r p m持續攪袢，噴霧溶融態之1 0重量份之牛脂衍生硬化 油（熔點4 8 °C )。溶融油之小滴溶解精細分割鯊魚軟骨 及形成基質。在此階段，然而，鯊魚軟骨表面部份露出且 包衣進行不充分。因此，每一初步基質的表面包覆以3 0 重量份之具有平均粒徑5微米之精細分割硬化菜油（熔點 6 8 °C )，其被選擇作爲不同熔點之脂質，經由裝入同一 攪拌製粒機及以攪拌速度約1 0 0 0 r p m攬拌該混合物 三十分鐘，藉以使該油配置及展開於初步基質表面。可因 此獲得具有充分廠酸性及持續釋放性質之非常滿意的油脂 基質包覆配方。 實施例1 經濟部中央標準局員工消費合作社印製 (請先閱讀背面之注意事項再填寫本頁) 老鼠內腫瘤血管生成抑制活性試驗 (對鯊魚軟骨粒徑之評價） 該試驗係經由背部氣囊法（d 〇 r s a 1 a i r s a c method)進行。一室經由使用具有直徑5 mm之兩.面塑膠 環之微過濾器而製備及注入該室內1 X 1 0 6MH— 1 3 4腫瘤細胞及培養基。老鼠被分成六組且將上述之室皮下 __15___ 本紙張尺度適用中國國家標準（CNS ) Λ4規格(210X 297公釐） ~ 577745 A7 B7 五、發明説明（κ) 植入每一隻老鼠之背部。 下述之鯊魚軟骨爲□服施藥，其劑量爲1 0 0 0毫克 /公斤，對四組老鼠施藥四天。在第五天，評佶血管生成 抑制程度。在一補充組（控制組）內，不施與鯊魚軟骨 且，以同樣的方法，在第五天評估抑制的程度。來自腫瘤 所漏出之血管生成促進物質透過微過濾器而造成新生血管 形成於與室接觸之表面。這些腫瘤誘導新生血管具有螺旋 狀形式β該血管生成之抑制程度根據以下之標準進行評 價。 (++) 3分：顯著之血管生成 (+ ) 2分：明確之血管生成 (± ) 1分：輕微之血管生成 (一）0分：無血管生成 抑制百分率以與控制組比較之分數減少率（％)表 7J\ 。 黨魚軟骨（1 ) : Cartilaid，爲一種美國黨魚軟骨產 經濟部中央標準局員工消費合作社印製 (請先閱讀背面之注意事項再填寫本頁) 品，被使用作爲鯊魚軟骨原料。該產物以液態氮（一 1 9 6 °C )瞬間冷凍及於截切碾磨機（定位碾磨機（Orient mill ))約略地粉碎。然後，仍然在冷凍的狀態中，以高 速旋轉式碾磨機（葉輪碾磨機（Impeller Mill))進一 步磨碎該粉碎物及最後以空氣一噴射式碾磨機（噴射碾磨 機（jet mill))磨碎。使用因此獲得之產物。 鯊魚軟骨（2 ):上述之鯊魚軟骨（1 )以3 2微米 篩子篩選及使用在篩子上之部份。 ___1Q_ 本紙張尺度適用中國國家標準（CNS ) A4規格（21〇X 297公釐) 577745 A7 B75. Description of the Invention (A) Film. The formulation can be extended and sustained release can be achieved by further coating or embedding the active substance in a polymer or wax. Such subcutaneous, intramuscular, intraarterial or intravenous administration can be performed by making a liquid unit dosage form, e.g., an injectable composition in solution or suspension. It can be manufactured by dissolving or suspending a certain amount of active substance in a non-toxic liquid carrier suitable for injection purposes, for example, an aqueous or greasy solvent, and sterilizing the resulting solution or suspension. It can also dispense a certain amount of active substance into a vial in powder or freeze-dried form and then sterilize and seal the vial and its contents. In this example, the vial and carrier are dissolved or mixed prior to application. Non-toxic salts or salt solutions can be added to make the injectable composition isotonic. In addition, a peripheral stabilizer, a preservative, a suspending agent, an emulsifier and / or the like may be added. The rectal dosage form can be produced by mixing the active substance with a hydrophobic or hydrophilic drug base, for example, polyethylene glycol, cocoa butter, higher esters (for example, myristyl palmitate) or a mixture thereof and kneading the The mixture is prepared. As described above, the tumor angiogenesis inhibitor of the present invention has no unpleasant taste and odor of shark cartilage, is acid-resistant and is only dissolved in the intestine, and can be conveniently administered by an oral route. In addition, it has reliable anticancer activity and analgesic activity and is very effective as a pharmaceutical composition. The combined use of the tumor angiogenesis inhibitor and the leukocyte interstitial- 12 inducer of the present invention shows very high anticancer activity and is therefore very effective as a pharmaceutical composition. The best mode for carrying out the invention _u___ This paper size applies Chinese National Standard (CNS) A4 specification (210X 297mm) " ~ ϋϋ r I-: I-I 1 2ϋ (Please read the notes on the back before filling (This page) Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 577745 B7 _ _ V. Description of the invention (0) The following production examples and examples are used to further illustrate the present invention in detail. They, however, in no way limit the scope of the invention. Production Example 1 Manufacturing of coated formula of shark cartilage-embedded oil or fat base. Fine-divided shark cartilage with an average particle diameter of 27 microns and 60 parts by weight was charged into a high-speed stirrer granulator (high-speed mixer, Shenjiang Industrial Co., Ltd.) And, keep the internal temperature of the granulator at 10 ° C and continuously stir at 20 to 100 rpm, spraying melted 10 parts by weight of tallow-derived hardened oil (melting point 4 8 ° C). The droplets of the melted oil dissolve the finely divided shark cartilage and form a matrix. At this stage, however, the surface of the shark cartilage was partially exposed and the coating was insufficient. Therefore, the surface of each preliminary matrix is coated with 30 parts by weight of a finely divided hardened vegetable oil (melting point 68 ° C) having an average particle diameter of 5 microns, which is selected as a lipid with a different melting point and prepared by loading the same agitation Granulator and stir the mixture for 30 minutes at a stirring speed of about 1000 rpm, so that the oil is deployed and spread on the surface of the preliminary substrate. This results in a highly satisfactory grease base coating formulation with full plant acidity and sustained release properties. Example 1 Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) Test of tumor angiogenesis inhibitory activity in mice (evaluation of shark cartilage particle size) This test is performed by the back airbag method (D rsa 1 airsac method). One chamber was prepared and injected into the chamber with 1 × 106MH—134 tumor cells and culture medium by using a microfilter with two-sided plastic rings with a diameter of 5 mm. The rats were divided into six groups and the above-mentioned chambers were subcutaneously __15___ This paper size applies the Chinese National Standard (CNS) Λ4 specification (210X 297 mm) ~ 577745 A7 B7 V. Description of the invention (κ) Implanted into the back of each mouse . The shark cartilage described below was administered at a dose of 1000 mg / kg and administered to four groups of mice for four days. On the fifth day, the degree of angiogenesis inhibition was evaluated. In a supplementary group (control group), shark cartilage was not administered and, in the same manner, the degree of inhibition was evaluated on the fifth day. An angiogenesis-promoting substance leaked from a tumor passes through the microfilter, causing new blood vessels to form on the surface in contact with the chamber. These tumor-induced neovascularization has a spiral form β. The degree of inhibition of angiogenesis is evaluated according to the following criteria. (++) 3 points: significant angiogenesis (+) 2 points: clear angiogenesis (±) 1 point: slight angiogenesis (1) 0 point: no angiogenesis inhibition percentage decreased as compared with the control group Rate (%) Table 7J \. Dangyu Cartilage (1): Cartilaid, printed by the Consumer Cooperative of the Central Standards Bureau of the Department of Economics, Dangyu Cartilage, U.S.A. (Please read the precautions on the back before filling this page) is used as a raw material for shark cartilage. The product was instantly frozen with liquid nitrogen (196 ° C) and roughly crushed in a cutting mill (Orient mill). Then, while still frozen, the pulverized material was further pulverized with a high-speed rotary mill (impeller mill) and finally an air-jet mill (jet mill) ))grinding. The product thus obtained is used. Shark cartilage (2): The above shark cartilage (1) is sieved through a 32-micron sieve and used on the sieve. ___1Q_ This paper size applies to Chinese National Standard (CNS) A4 (21〇X 297 mm) 577745 A7 B7
五、發明説明（J 鯊魚軟骨（3 ):上述之鯊魚軟骨（1 篩子篩選及使用通過篩子之部份。 結果如表1所示。 表1 以3 2微米 Η~ ~h (3分） + (2分） 土 (1分） (0分） 搬 控制組 鯊魚軟骨(1) 鯊魚軟骨I(2) 鯊魚軟骨 (3) 4隻麵 2隻 2隻 1集 2隻老鼠 0隻老鼠 om 0隻老鼠 2·7土 0.5 2·2土0.8 18.5 0集 0隻親 2.3土 0.5 14.8 .--1----0 — I (請先閱讀背面之注意事項再填寫本頁) 4隻老鼠 im om 2.〇士 0.8 25.9 *Ρ<0.05~ 上述結果指出，在測試之鯊魚軟骨粉末中，通過3 2 微米篩子之精細分割鯊魚軟骨顯示出明顯高度血管生成抑 制活性。 實施例2 老鼠內腫瘤血管生成抑制活性試驗 (Cartilaid 對 Bettershark) 以如實施例1之同樣方法使用背部氣囊法比較美國黨 魚軟骨Cartilaid及藍鯊衍生軟骨Bettershark。結果如表 2所示。 在1 0 0毫克/公斤C a r t i 1 a i d組中，抑制百分率（與 控制組比較之分數減少率（％ ))爲3 · 6 %但，在1 〇 0 0毫克/公斤Cartilaid組中，其爲2 1 ♦ 4%具有顯著赛 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） 訂 -tf! 經濟部中央標準局員工消費合作社印製 577745 A7 B7 五、發明説明（|k ) 異（Ρ<0·05)。反之，在 Bettershark組中，在 1〇 0毫克/公斤組中之抑制百分率爲1 7 · 9 %具有顯著差異 (P < 0 . 0 5 )且在1 〇 〇 〇毫克/公斤組中之抑制百分 率爲高達3 5.7%，爲統計上明顯差異（P<0.0 〇 2)。在〇31：1丨13丨(1與66 1“以1^^間，亦，發現明顯差異 (P < 0 · 0 5 )。 表2 劑量（毫 克/公 斤） 1 ++ (3分） + (2分） 土 (1分） (0分） 分數 mm 控制組 — 9隻親 1隻雜1 0隻纖 0隻纖 2.8 土 0.4 Cartilaid 1000 3mm 6隻纖 imM, 0隻老鼠 2.2± 0.8 21.4* Caitilaid 100 6隻纖 4隻雜 〇隻纖 0隻老鼠 2.7± 0.5 3.6 Better — shark 1000 3隻老鼠 2mm 5隻老鼠 0隻老鼠 1.8± 1.0 35.7* 氺 Better — shade 100 5隻親 4隻織 1隻纖 0隻親 2.3土 0.8 17.9* *Ρ<〇·〇5，"Ρ<0·002。 上述結果顯示Bettershark比Cartilaid具有較高之腫V. Description of the invention (J Shark cartilage (3): The above mentioned shark cartilage (1 sieve screened and used through the sieve. The results are shown in Table 1. Table 1 with 3 2 micron Η ~ ~ h (3 points) + (2 points) Soil (1 point) (0 points) Shark cartilage (1) Shark cartilage I (2) Shark cartilage (3) 4 noodles 2 2 1 set 2 mice 0 mouse om 0 Rat 2 · 7 soil 0.5 2 · 2 soil 0.8 18.5 0 episodes 0 pro 2.3 soil 0.5 14.8 .-- 1 ---- 0 — I (Please read the precautions on the back before filling this page) 4 mice im om 2.00 0.8 25.9 * P < 0.05 ~ The above results indicate that in the shark cartilage powder tested, the finely divided shark cartilage passed through a 32 micron sieve showed a significantly high angiogenesis inhibitory activity. Example 2 Tumor angiogenesis in mice Inhibitory activity test (Cartilaid vs. Bettershark) The same method as in Example 1 was used to compare the US party fish cartilage Cartilaid and Blue Shark-derived cartilage Bettershark using the back airbag method. The results are shown in Table 2. At 100 mg / kg Carti 1 In the aid group, the percentage of inhibition (fraction reduction rate (%) compared with the control group) was 3 · 6%, but in the Cartilaid group of 1000 mg / kg, it is 2 1 ♦ 4% has a significant paper size Applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) Order -tf! Ministry of Economic Affairs Printed by the Consumer Standards Cooperative of the Central Bureau of Standards 577745 A7 B7 V. Description of the Invention (| k) Different (P < 0.05). Conversely, in the Bettershark group, the inhibition percentage in the 100 mg / kg group was 17 9% had a significant difference (P < 0.05) and the percentage inhibition was as high as 3 5.7% in the 1000 mg / kg group, which was a statistically significant difference (P < 0.02). 31 ： 1 丨 13 丨 (1 and 66 1 "with 1 ^^, also, significant differences were found (P < 0 · 0 5). Table 2 Dose (mg / kg) 1 ++ (3 points) + ( 2 points) Soil (1 point) (0 points) Score mm Control group — 9 pros, 1 miscellaneous, 1 fiber, 0 fiber, 2.8 soil, 0.4 Cartilaid 1000 3mm, 6 fiber imM, 0 mouse 2.2 ± 0.8 21.4 * Caitilaid 100 6 fibers 4 hybrids 0 fibers 0 mice 2.7 ± 0.5 3.6 Better — shark 1000 3 mice 2mm 5 mice 0 mice 1.8 ± 1.0 35.7 * etterBetter — shade 100 5 pros, 4 weaving, 1 fiber, 0 pros, 2.3 soil 0.8 17.9 * * P < 〇 · 05, " P < 0 · 002. The above results show that Bettershark has a higher swelling than Cartilaid
(請先閲讀背面之注意事項再填寫本頁) 瘤血管生成抑制活性。 經濟部中央標準局員工消費合作社印製 實施例3 老鼠內腫瘤生長抑制試驗（B e 11 e Γ s h a r k有效劑量發現之 試驗） 腫瘤細胞（肉瘤1 8 Ο ) ，1 x 1 〇 6，皮下植入二 組中之每一隻老鼠（I c R老鼠.，雄性，六週大）的背 部，每一組有十隻。在一組中，立即在其後，以1 0 0 0 ____18 __ —- 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） 577745 經濟部中央標準局員工消費合作社印製 Λ7 B7 五、發明説明（Π ) 毫克/公斤之劑量一天一次口服施與(:81:1:1131(1 (美國產 品）二十天。另一組’以同樣劑量同樣期間施與 Bettershark (具有調整至不大於3 2微米之粒徑；確定 在P Η値2之三小時期間能穩定）°在第三組’動物餵養 二十天不施與任何藥物。在經過二十五天後，殺死所有動 物及將腫瘤稱重。獲得之結杲如表3所示。抑制百分率以 與控制組比較之腫瘤重量減少百分率顯示。然而，在 C a r t i i a i d組中，與控制組比較之抑制百分率爲1 1 . 1 %，在Bettershark組中之抑制百分率爲4 7 · 0%具有 顯著差異（Ρ<〇·〇〇1)。在 Cartilaid 與 Bettershark間之比較顯示Bettershark具有較高之抗 癌活性（P < 0 · 0 5 )。 表3 平均驄t*量（克） 抑制％ 控制組 2.752±1·388 一 Caitilaid 2·446±1·1 39 11.1* Bettashatk 1·457±1·965 47.0** *Ρ<0·05，"Ρ<0· D01 上述之結果建議Bettershark (具有不大於3 2微米之 粒徑），每個個體每天劑量2 0克，在臨床實務上將足夠 有效。 實施例4 老鼠內止痛效果試驗 經由醋酸扭轉法（acetic acid writhing method) 評估Bettershark之止痛效果。I C R老鼠（雄性，五週 19 本紙張尺度適用中國國家標準（CNS ) Λ4規格（210 X 297公釐） (請先閱讀背面之注意事項再填寫本頁) 訂 577745 A7 B7 五、發明説明（) 大）分成五組，每一組有十隻。根據下列所述之表對每一 組施與每一種測試物質且，在經過四十天後，腹膜內施與 2 %醋酸。在施與醋酸後自十天至二十天期間，計算每一 隻老鼠之扭轉反射。在施與醋酸後，每天施與測試物質直 到扭轉反射計算終止。 在Cartilaid組中，口服施與每天劑量1 0 0 0毫克/ 公斤之Cartilaid ;在Bettershark組中，口服施與每天劑 量1 0 0 0毫克/公斤之Bettershark ;在Indacin組 中，口服施與劑量2 5毫克/公斤之Indacin (消炎痛，非 類固醇止痛；商業產品）；在硫酸軟骨素組中，口服施與 劑量3 3 3毫克/公斤之硫酸軟骨素（商業產品）；及， 在控制組中，餵養之動物不施與任何藥物β抑制百分率以 與控制組比較之該反射次數減少百分率表示。結果如表4 所示。同樣地關於止痛效果，Bettershark之優越性可因 而被建立。 表4 mm^rn 「麵％ 控制組 3 1·2±9·6 — Cartilaid 24·3±3.9 23.6* Bdlershaik 21·2±10·4 32.1* Indadn 1 0·3±5·6 67.0** 23·4±4·5 25.0* *Ρ<0·05，**Ρ<0·( D01 經濟部中央標準局員工消費合作社印製 (請先閱讀背面之注意事項再填寫本頁) 實施例5 免疫專一性抗原在B e 11 e r s h a r k施.藥之臨床病例的表現 20 本紙張尺度適用中國國家標準（CNS ) Α4規格（210X 297公釐） 577745 Λ7 B7 _ 五、發明説明（θ ) 一個8 0歲男性抱怨其上腹部有不快感。基於胃鏡檢 查發現，其被診斷出在胃角上部位（胃角切迹）具有I I c進行型胃癌（印戒細胞癌）。使用活體組織檢查樣本作 電子顯微鏡及組織檢查（免疫檢查）。他因爲心肌梗塞而 不宜動手術。 使用於實施例1中之鯊魚軟骨（3 )(此後，在實施 例5中，稱爲“Bettershark” ）施與上述病患每天每日 劑量2 0克。電子顯微鏡及免疫特殊染色（第一次胃鏡檢 查）在Bettershark施藥前僅有通常癌細胞之一般的發 現，脫噬作用指示之發現非常輕微。沒有觀察到粘連因子 (螢光抗體法）或F a s抗原（特殊染色）及沒有觀察到 HSP6 0 (應激性蛋白，熱休克蛋白；螢光抗體法）之 表現。在第一次胃鏡檢查二個半月後進行之胃鏡檢查發現 指出癌的周圍明顯地被正常黏膜所覆蓋，周圍癌突出其顯 示出傾向平坦的趨勢。電子顯微鏡發現在第二次活體組織 檢查中顯示細胞膜及細胞質沒有特別改變但，淸楚地顯示 癌細胞之狹縮及分裂，可觀察到脫噬體。即，獲得於癌細 胞脫噬作用之陽性發現指示。 經濟部中央標準局員工消費合作社印製 (請先閱讀背面之注意事項再填寫本頁) 在此時期，活體組織檢查樣本進行免疫特殊染色。經 由使用抗HSP60抗體，抗CD80抗體，抗CD86 抗體及，此外，抗F a s抗體檢查癌細胞。施與 Bettershark配方之結果，(^— 原成陽性且，此外， HSP 6 0抗原及CD80及CD 8 6之粘連因子成陽 性。 2J____ 本紙張尺度適用中國國家標準（CNS ) A4規格（210X 297公發) ^ 577745 A7 經濟部中央標準局員工消費合作社印製 __B7 _五、發明説明（>。） 在第三次胃鏡檢查（約在第一次胃鏡檢查五個月後） 中，在上部胃角之癌已完全地消失。胃癌部位之活體組織 檢查樣本已被發現對癌細胞皆呈陰性。 •上述之發現摘述如下。F a s抗原，HSP 6 0抗原 及C D 8 0及C D 8 6之粘連因子，其在Bettershark配 方施與之前沒有被觀察到，在施與Betters hark配方二個 半月皆成陽性。其後，第三次胃鏡檢查顯示癌已完全消 失。此現象可由下列所解釋。經由Bettershark配方之施 與使供給腫瘤之血液被抑制且，因此，應激性蛋白（HS P60)被表現，及Fas抗原及CD80及CD86之 粘連因子呈陽性。其後，在第三次胃鏡檢查中，證實癌完 全退化。 腫瘤免疫學之硏究已顯示腫瘤退化包含壞死及脫噬。 壞死爲由非特異的免疫作用之發炎或其他因子所導致之腫 瘤消失現象，其中腫瘤細胞被破壞爲以順序發生在細胞 膜，細胞質及核變更之結果。組織學地，在壞死的例子 中，其發生嗜中性白血球，巨噬細胞及纖維組織母細胞之 浸潤，其中，其發生石灰化及纖維生成，造成鹽細胞之消 失。因此，在此位置仍有痕跡殘留。 另一方面，在源自脫噬之腫瘤消失的例子中，沒有發 炎產生及沒有痕跡殘留但正常組織被發現取代。其已知， 在此例子中，殺傷性T細胞識別在腫瘤細胞表面之F a s 抗原及對其黏著且與粘連因子如C D 8 0或/及C D 8 6 共同作用同時地誘導脫噬作用。核變質（核分裂及脫噬體 ____22____ 本紙張尺度適用中國國家標準（CNS ) A4規格（210X 297公浚) ' (請先閱讀背面之注意事項再填寫本頁) 577745 A7 B7 五、發明説明（ >丨） 形成）首先發生及然後細胞質及細胞膜被巨噬細胞吞_ ° 因此，沒有痕跡殘留。 在上述及其他Bettershark配方之施藥的例子中’在 癌消失部位並沒有獲得例如，石灰化或纖維生成等發炎痕 跡之暗示發現及，因此，其暗示癌之消失應是由於脫π遼作 用。其顯然在非常短的期間（一個月至一個半月）該癌已 經消失。 實施例6 老鼠內免疫專一性試驗 經濟部中央標準局員工消費合作社印製 (請先閱讀背面之注意事項再填寫本頁) 肉瘤1 8 0腫瘤細胞植入每一隻老鼠的背部。比較腫 瘤生長抑制效果，其中一組腹膜內施與1 0毫克ΤΝΡ 4 7 0 (武田藥品工業社製），其中一組口服施與1 〇 〇 〇 毫克/公斤之Bettershark，其中一組腹膜內施與2 4微 克angiostatin (化學及血淸療法硏究所製）及控制組不施 與藥劑。在二星期及三星期後，在所有劑量組中發現腫瘤 生長被抑制及與控制組相比具有顯著差異（Ρ < 〇 . 〇 5)。在這時期免疫組織檢查該腫瘤組織之HSP60, Fa s抗原，CD80及CD86之表現，及發現HSP 60，Fa s抗原，CD80及CD86之表現程度明顯 地增加。 個別地，使用缺少T細胞老鼠，即裸鼠，進行上述同 樣的實驗。每一藥劑顯示腫瘤生長抑制作用於特定範圍但 沒有任何顯著差異。根據腫瘤組織之免疫組織檢查，發現 HSP60，Fa s抗原，CD80及CD86之表現爲 ____23__ 本紙張尺度適用中國國家標準（CNS ) A4規格（210X 297公釐） 57^745 經濟部中央標準局員工消費合作社印製 A7 B7_ 五、發明説明（/) 陽性。然而，並沒有獲得任何暗示脫噬之發現。 上述結果暗示血管生成抑制劑之抗腫瘤活性並不只基 於僅停止血液供應至腫瘤但亦在於殺傷性τ細胞識別在抑 制血液烘應至腫瘤的狀態中應激性蛋白表現或在F a s抗 原，C D 8 0及C D 8 6表現中之定性及定量之改變，其 中，且展示它們的抗腫瘤活性，藉此誘導脫噬作用。 實施例7至1 0 AHC C及Bettershark合倂施與之病例 表5所示之四個癌症病患，每天施與每日劑量3 · 0克 之AH C C且同時每天口服施與每日劑量2 0克之 Bettershark。在施與三個月後，評估治療結果及測定N K活性，白血球間質一 1 2數値及C D 4 / C D 8値。在 實施例7及1 0中，腫瘤縮小5 0 %或以上。N K活性， 然而，仍然在正常範圍內，並沒有顯示任何顯著活性增 加。在每一實施例7至1 0中，白血球間質一 1 2數値顯 然地高於正常範圍。因而，可推測白血球間質一 1 2包含 在腫瘤減小作用內。 在表5中，CD4爲輔助T細胞標誌，及CD8爲殺 傷性T細胞標誌，及C D 4 / C D 8値反映殺傷性T細胞 數目增加的範圍。小於1之數値表示殺傷性T細胞數目增 力口。其正常範圍是1.0至1.5。 24 本紙張尺度適用中國國家標準（CNS ) A4規格（210X29*7公釐） (請先閱讀背面之注意事項再填寫本頁)(Please read the precautions on the back before filling out this page) Tumor angiogenesis inhibitory activity. Printed in Example 3 by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs, a tumor growth inhibition test in mice (a test for the effective dose of Be 11 e Γ shark) Tumor cells (sarcoma 1 8 〇), 1 x 10, subcutaneously implanted There are ten mice in each group on the back of each mouse (IcR mouse., Male, six weeks old). In one group, immediately after this, 1 0 0 0 ____18 __ —- This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) 577745 Printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs Λ7 B7 5 2. Description of the invention (Π) mg / kg orally administered once a day (: 81: 1: 1131 (1 (US product) for twenty days. Another group was administered to Bettershark at the same dose and the same period (with adjustment to not Particle size greater than 32 microns; determined to be stable during three hours of P Η 値 2) ° In the third group, animals are fed for 20 days without any medication. After 25 days, all animals are killed The tumors were weighed. The obtained scabs are shown in Table 3. The percentage of inhibition is shown as a percentage reduction in tumor weight compared to the control group. However, in the Cartiiaid group, the percentage of inhibition compared to the control group was 1.1. 1 %, The percentage of inhibition in the Bettershark group was 47. 0% with a significant difference (P < 〇〇〇〇1). A comparison between Cartilaid and Bettershark showed that Bettershark has a higher anticancer activity (P < 0 · 0 5). 3 Mean 骢 t * amount (g) Inhibition% Control group 2.752 ± 1 · 388-Caitilaid 2.446 ± 1 · 1 39 11.1 * Bettashatk 1.457 ± 1 · 965 47.0 ** * P < 0 · 05, " P < 0 · D01 The above results suggest that Bettershark (with a particle size of not more than 32 microns), a daily dose of 20 grams per individual, will be effective in clinical practice. Example 4 Test of analgesic effect in rats was reversed by acetic acid Method (acetic acid writhing method) to evaluate the analgesic effect of Bettershark. ICR mice (male, 19 weeks, 5 weeks) This paper size applies the Chinese National Standard (CNS) Λ4 specification (210 X 297 mm) (Please read the notes on the back before filling This page) Order 577745 A7 B7 V. Description of the invention () Large) is divided into five groups of ten. Each test substance is administered to each group according to the table described below, and after forty days have passed. 2% acetic acid was administered intraperitoneally. The twisted reflex of each mouse was calculated from 10 to 20 days after the acetic acid was administered. After the acetic acid was applied, the test substance was administered daily until the twisted reflex calculation was terminated. In the Cartilaid group, Cartilaid is administered daily at a dose of 1000 mg / kg; Bettershark is administered orally in the Bettershark group; Bettershark is administered orally at a dose of 100 mg / kg daily; in the Indacin group, it is administered orally at a dose of 25 mg / kg Indacin (indomethacin, non-steroidal analgesic; commercial product); in the chondroitin sulfate group, chondroitin sulfate (commercial product) was administered orally at a dose of 33 mg / kg; and, in the control group, the animals fed were not The percentage inhibition of any drug administered was expressed as a percentage reduction in the number of reflections compared to the control group. The results are shown in Table 4. As for the analgesic effect, Bettershark's superiority can therefore be established. Table 4 mm ^ rn "Face% control group 3 1 · 2 ± 9 · 6 — Cartilaid 24 · 3 ± 3.9 23.6 * Bdlershaik 21 · 2 ± 10 · 4 32.1 * Indadn 1 0 · 3 ± 5 · 6 67.0 ** 23 · 4 ± 4 · 5 25.0 * * P < 0 · 05, ** P < 0 · (D01 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) Example 5 Immunization Performance of specific antigens in clinical cases administered by Be 11 ershark. 20 This paper size applies the Chinese National Standard (CNS) A4 specification (210X 297 mm) 577745 Λ7 B7 _ V. Description of the invention (θ) A 80 years old The male complained of unpleasantness in his upper abdomen. Based on gastroscopy, he was diagnosed with type II c progressive gastric cancer (signet ring cell carcinoma) in the upper part of the stomach corner (stomach corner notch). Biopsy samples were used for electron microscopy and Histological examination (immunological examination). He is not suitable for surgery due to myocardial infarction. The shark cartilage (3) used in Example 1 (hereinafter, referred to as "Bettershark" in Example 5) was administered to the above patients every day Daily dose of 20 grams. Electron microscope and special immunostaining (section Sub-gastroscopy) There were only common findings of cancer cells before Bettershark administration, and the discovery of phagocytosis was very slight. No adhesion factor (fluorescent antibody method) or Fas antigen (special staining) was observed and no observation To HSP60 0 (stress protein, heat shock protein; fluorescent antibody method). Gastroscopy performed two and a half months after the first gastroscopy revealed that the surrounding area of the cancer was clearly covered by normal mucosa, and the surrounding cancer It shows that it tends to be flat. Electron microscopy showed that the cell membrane and cytoplasm were not particularly changed in the second biopsy. However, it clearly showed the narrowing and division of cancer cells, and dephagosomes could be observed. That is, Obtained a positive detection indication for cancer cell phagocytosis. Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling out this page). During this period, biopsy samples were subjected to special immunostaining. Anti-HSP60 antibody, anti-CD80 antibody, anti-CD86 antibody and, in addition, anti-Fas antibody to check cancer cells. Administration to Bett The results of the ershark formula, (^-original positive and, in addition, HSP 60 0 antigen and CD80 and CD 86 6 adhesion factors were positive. 2J____ This paper size applies Chinese National Standard (CNS) A4 specifications (210X 297) 577745 A7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs __B7 _ V. Description of Invention (>. In the third gastroscopy (about five months after the first gastroscopy), the cancer in the upper gastric horn has completely disappeared. Biopsy samples from gastric cancer sites have been found to be negative for cancer cells. • The findings above are summarized below. Fas antigen, HSP 60 antigen, and CD 80 and CD 86 adhesion factors were not observed before the administration of Bettershark formula, and were positive for two and a half months after the administration of Bettersark formula. Thereafter, a third gastroscopy revealed that the cancer had completely disappeared. This phenomenon can be explained as follows. The administration of the Bettershark formula inhibited blood supply to the tumor and, therefore, stress protein (HS P60) was expressed, and Fas antigen and CD80 and CD86 adhesion factors were positive. Thereafter, in the third gastroscopy, it was confirmed that the cancer had completely degenerated. Research in tumor immunology has shown that tumor regression includes necrosis and phagocytosis. Necrosis is the disappearance of tumors caused by non-specific immune-induced inflammation or other factors, in which tumor cells are destroyed as a result of sequential changes in the cell membrane, cytoplasm, and nucleus. Histologically, in the case of necrosis, it occurs infiltration of neutrophils, macrophages, and fibroblasts. Among them, calcification and fibrogenesis occur, resulting in the loss of salt cells. Therefore, traces remain at this location. On the other hand, in the case of the disappearance of the tumor from dephagy, no inflammation was generated and no trace remained but normal tissue was found to be replaced. It is known that, in this example, killer T cells recognize the Fas antigen on the surface of tumor cells and adhere to it and co-act with adhesion factors such as CD80 or CD50 and induce dephagy simultaneously. Nuclear Metamorphism (Nuclear fission and phagosome ____22____ This paper size applies Chinese National Standard (CNS) A4 specification (210X 297 Gong Jun)) (Please read the precautions on the back before filling this page) 577745 A7 B7 V. Description of the invention ( > 丨) Formation) occurs first and then the cytoplasm and cell membrane are swallowed by macrophages. Therefore, no trace remains. In the above and other examples of the application of the Bettershark formula, 'the hint that no signs of inflammation such as calcification or fibrogenesis have been obtained at the site where the cancer disappeared, and therefore, it suggests that the disappearance of the cancer should be due to the effect of de-πliao. It was clear that the cancer had disappeared in a very short period of time (one to one and a half months). Example 6 Immune Specificity Test in Rats Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling out this page) Sarcoma 180 tumor cells were implanted into the back of each mouse. To compare the tumor growth inhibitory effects, one group was administered intraperitoneally with 10 mg of TNP 47 (manufactured by Takeda Pharmaceutical Industry Co., Ltd.), one group was administered orally with 1,000 mg / kg of Bettershark, and one group was administered intraperitoneally. And 2 4 μg angiostatin (manufactured by the Chemical and Hemostasis Therapy Laboratory) and the control group did not administer any medication. After two and three weeks, tumor growth was suppressed and significantly different from the control group in all dose groups (P < 0.05). During this period, the immune tissues examined the expression of HSP60, Fas antigen, CD80 and CD86 in the tumor tissue, and found that the expression of HSP60, Fas antigen, CD80 and CD86 increased significantly. Individually, the same experiment was performed using a T cell-deficient mouse, that is, a nude mouse. Each agent showed tumor growth inhibition in a specific range without any significant difference. According to the immune tissue examination of the tumor tissue, it was found that the performance of HSP60, Fas antigen, CD80 and CD86 is ____23__ This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X 297 mm) 57 ^ 745 Employees of the Central Standards Bureau of the Ministry of Economic Affairs Printed by Consumer Cooperative A7 B7_ V. Invention Description (/) Positive. However, no findings suggestive of phagocytosis were obtained. The above results suggest that the anti-tumor activity of angiogenesis inhibitors is not only based on stopping blood supply to tumors only, but also on the killer τ cells recognizing stress protein expression in inhibiting blood-to-tumor states or on Fas antigen, CD Qualitative and quantitative changes in the expression of 80 and CD 86, among which they exhibit their antitumor activity, thereby inducing dephagocytosis. Examples 7 to 10 Cases in which AHC C and Bettershark were administered in combination The four cancer patients shown in Table 5 were administered a daily dose of 3.0 g of AH CC daily and were administered orally daily at a daily dose of 20 Bettershark. Three months after the administration, the treatment results were evaluated and the NK activity was measured. The leukocyte interstitial count was 12 and the CD 4 / CD 8 were measured. In Examples 7 and 10, the tumor shrank by 50% or more. NK activity, however, was still in the normal range and did not show any significant increase in activity. In each of Examples 7 to 10, the leukocyte interstitial number is significantly higher than the normal range. Therefore, it can be presumed that leukocyte mesenchyme-1 2 is included in the tumor reducing effect. In Table 5, CD4 is a helper T cell marker, CD8 is a killer T cell marker, and CD 4 / CD 8 (値) reflects the range of increase in the number of killer T cells. A number less than 1 indicates an increase in the number of killer T cells. Its normal range is 1.0 to 1.5. 24 This paper size applies to China National Standard (CNS) A4 (210X29 * 7mm) (Please read the precautions on the back before filling this page)
577745 A7 B7 五、發明説明（β ) 經濟部中央標準局員工消費合作社印製 表5 實麵 病患 NK活注 血淸中之白 CD4/C 血·質一 D8 12 7 72歲，男 編生胃癌 24% 2 4 0 π g CR | 0.34 性 /ml 1 ....二 8 5了歲，男 盲 62% 2 3 0 n g CR 0.42 性 癌性愎麟 /ml 9 69歲，男 胃癌 6 2% 1 0 3 n g CR 0.62 性 mm /ml 10 71歲，女 肝癌 68% 7 8 n g/ PR 0.84 性 wm 纖 m 1 CR :完麵七 PR :咨讓化 (請先閱讀背面之注意事項再填寫本頁) 本紙張尺度適用中國國家標準（CNS ) A4規格（210X 297公釐）577745 A7 B7 V. Description of the invention (β) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. 5 Real patients NK live injection of white CD4 / C in blood. Blood quality D8 12 7 72 years old, male editor Gastric cancer 24% 2 4 0 π g CR | 0.34 Sex / ml 1 .... 2 8 5 years old, male blind 62% 2 3 0 ng CR 0.42 Sexual cancerous lin / ml 9 69 years old, male gastric cancer 6 2 % 1 0 3 ng CR 0.62 Sex mm / ml 10 71 years old, female liver cancer 68% 7 8 ng / PR 0.84 Sex wm Fiber m 1 CR: Finish 7 PR: Consultation (please read the precautions on the back before filling (This page) This paper is sized for China National Standard (CNS) A4 (210X 297mm)