471972 AT —_ B7五、發明説明（1 ) 經濟部中央標率局員工消費合作社印製 發明領域 本發明係關於一種紫杉烷類組成物，尤指一種適於靜 脈注射之醫藥用紫杉烷類組成物。 發明背景 紫杉醇（Taxol®)，亦稱作太平洋紫杉醇（paclitaxeO，是一種 由西方紫杉(Taxus brevifolia)樹皮中萃取而得的化合物。紫杉醇 以作爲抗癌藥劑使用而知名。以紫杉醇治療那些經cisplatin 或長春花屬生物鹼（vinca alkaloid)治療無效的卵巢癌和乳癌病 人，顯示有良好的反應。紫杉醇亦被硏究用來治療各種其 它的癌症，如黑色素瘤（melanoma)、淋巴瘤（lymphoma)和肺 癌。 設計紫杉醇劑型的主要困難處在於其於水溶液中之溶 解度低。此因紫杉醇缺乏在藥理上可接受之範圍內能夠離 子化的官能基，而控制改變其pH値並無法增加其溶解性， 做成鹽類或加入帶電荷之螯合劑亦不適用（R. M. Straubinger， ”Taxol: Science and App丨ications”（M. SufTness編輯），C RC 刊印， New York，NY，第237-239頁，1 9 9 5 )。因此，如何將紫杉醇 與具生體可溶性之載送體製成配方，成爲其治療發展上的 一項挑戰。 過去在改善紫杉醇劑型溶解性和毒性的過程中，一些 藥物載送體（vehicles)曾被硏究過。一般而言，這些載送體包 括一些溶劑，如乙醇、二甲亞4風（dimethylsulfoxide)或低分子量 的聚乙烯醇（例如P E G 4 0 0 )。部分劑型中添加了油類或 ___2 本紙張尺度適用中國國家標率（CNS ) A4規格（210X297公釐） (請先閱讀背面之注意事項再填寫本頁) !·. 装！·471972 AT —_ B7 V. Description of the invention (1) Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economics Field of Invention The present invention relates to a taxane composition, especially a pharmaceutical taxane suitable for intravenous injection Class composition. BACKGROUND OF THE INVENTION Taxol®, also known as paclitaxeO, is a compound extracted from the bark of Taxus brevifolia. Paclitaxel is well known for its use as an anticancer agent. Paclitaxel is used to treat those treated with cisplatin Or vinca alkaloid has not responded well to patients with ovarian and breast cancer. Paclitaxel has also been studied to treat various other cancers, such as melanoma and lymphoma. And lung cancer. The main difficulty in designing a paclitaxel formulation is its low solubility in aqueous solutions. This is because paclitaxel lacks functional groups that can be ionized within a pharmacologically acceptable range, and controlling changes in its pH 値 does not increase its solubility It is also not suitable to make salts or add charged chelating agents (RM Straubinger, "Taxol: Science and App 丨 ications" (edited by M. SufTness), printed by C RC, New York, NY, pp. 237-239, 1 9 9 5). Therefore, how to formulate paclitaxel with a carrier that is soluble in organisms has become its therapeutic development A challenge in the past. Some drug carriers have been investigated in the past to improve the solubility and toxicity of paclitaxel dosage forms. Generally, these carriers include solvents such as ethanol, dimethylamine 4 wind (dimethylsulfoxide) or low molecular weight polyvinyl alcohol (such as PEG 4 0 0). Oil or ___2 is added to some dosage forms. The paper size is applicable to China National Standard (CNS) A4 specification (210X297 mm) (please (Please read the notes on the back before filling out this page)!
In n PI! 471972 A7 __ —— —________ B7五、發明説明（2 ) 經濟部中央標隼局員工消費合作社印製 界面活性劑，例如一種聚氧乙烯山梨醇之脂肪酸酯（亦即 "TWEEN 80”，亦稱作p〇lySOrbate)，聚乙氧基化之篦麻油（亦 即”CreniophorEL”），大豆油或三醋酯《但是這些劑型不是 遭遇到低溶解性（特別是在生理食鹽水中稀釋以進行靜脈 注射時），就是遭遇到高毒性的問題。高毒性的問題是由 油類或界面活性劑所引起。特別是當施用”TWEEN 80”之濃度 高到足以溶解高濃度之紫杉醇時，臨床上會引起胸膜積液 (pleural effusions)及水腫（edema)，而”Cremophor EL”則可導致嚴重 甚至致命的過敏（hypersensitivity)反應（R. M. Straubinger，"Taxol: Science and Applications”（M. Stiffness 編輯}，C R C 刊印，New York，NY，第 241 及 244頁，1 9 9 S )。 因此，亟需改善紫杉醇之劑型，使其在保有長期儲存 之穩定性的前提下，能具有低毒性。 發明槪述 本發明之目的，係提供一種紫杉烷類組成物，其因安 定性高、毒性低而適於靜脈注射之醫藥用途，可用以阻止 生體癌症之繼續成長^ 本發明之-方面係關於一種紫杉烷類組成物，其係將 紫杉烷類化合物儲存於一具有較佳溶解度和較低毒性之儲 存溶液中而成，其中該儲存溶液包括：（a)聚氧乙烯山梨 醇之脂肪酸單酯；（b)聚乙氧基化之篦麻油；以及（c>乙 醇；在此溶液中，單酯和聚乙氧基化之篦麻油共存，其 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） (請先閱绩背面之注意事項再填寫本頁) :# ............................................. 0 471972 A7 B7 ,^ ..................... ..... LUI. 11«»"——r —I---1 — .............................................-..........._l I ΙΙ.·»·_·.Ι’τ._..·Ι ..1,.ΙΙ.ΙΙΙΙ.ΊΙ.! ......................u,„ ^ _上.五、發明説明（3 ) 經濟部中央標隼局員工消費合作社印製 量足以有效降低紫杉烷類組成物之毒性，且該儲存溶液之 p Η値以在約1 ~ 8之間爲佳。 其中’該聚氧乙烯山梨醇之脂肪酸單酯較佳爲聚氧乙 烯（20)山梨醇單油酸酯；而該儲存溶液較佳更包括低分子 量之聚乙烯乙二醇，例如FEG 30 0 »於一較佳實施例中， 該紫杉烷類組成物包含4毫克/毫升至8毫克/毫升之紫杉 烷類化合物、20至30 % (ν/ν)聚乙氧基化篦麻油、s至 lS%(v/v)聚氧乙烯（20)山梨醇單油酸酯、15至 3❹％ ( v / v >乙醇和4 0至6 0 % ( v / v >之低分子量聚乙烯乙二 醇。 其中，該儲存溶液較佳更包括-·種酸當作緩衝劑，而 使其中的p Η値保持在大約4到6之間。該酸可爲無水檸檬 酸。 其中，該紫杉烷類化合物可爲紫杉醇或紫杉德 (decetaxel) ° 本發明之另一方面係關於一種適於靜脈注射之醫藥用 紫杉烷類組成物，該紫杉烷類組成物係將紫杉烷類化合物 儲存於一具有較佳溶解度和較低毒性之儲存溶液中，再以 一適於靜脈注射之稀釋劑稀釋而成，其中該儲存溶液包 括：（a>聚氧乙烯山梨醇之脂肪酸單酯；（b>聚乙氧基化之 篦麻油；以及（c)乙醇；在此溶液中，單酯和聚乙氧基化之 篦麻油共存，其量足以有效降低紫杉烷類組成物之毒性， 且該儲存溶液之pH値以在約1〜8之間爲佳。 其中，該稀釋劑係生理食鹽水。 4 本紙張尺度適用中國國家標隼（CNS ) A4規格（210X297公釐） (讀先閱讀背面之注意事項再填寫本頁) if 裝--- 88»:'· - 1Β 8ϋ 訂 tor • m -ml f «i 8*1 ί_1 I, : . SI > 471972 A7 _ ——— B7 五、發明説明（4 ) 經濟部中央標準局員工消費合作社印製 本發明之又一方面係關於一種利用紫杉烷類組成物治 療哺乳類動物癌症之方法，包括將一適於靜脈注射之醫藥 用紫杉烷類組成物以醫藥上可接受之量注射入生體中之步 驟’其中該紫杉烷類組成物係將紫杉烷類化合物儲存於一 具有較佳溶解度和較低毒性之儲存溶液中，再以一適於靜 脈注射之稀釋劑稀釋而成，其中該儲存溶液包括：（3>聚 氧乙烯山梨醇之脂肪酸單酯；（b)聚乙氧基化之篦麻油； 以及 (C>乙醇；在此溶液中，單酯和聚乙氧基化之篦麻油共存， 其量足以有效降低紫杉烷類組成物之毒性，且該儲存溶液 之P Η値以在約1 ~8之間爲佳。 本發明的其它目的及特性將在下文中詳述之。 本發明之詳細說明 I ·定義 以下所用術語，其意義如下： ”紫杉烷類”意指任何可抑制微小管之去聚合 (depo丨ymerization)之具有紫杉醇6-8-6稠環（fused ring)骨架之化合 物’該骨架包括紫杉醇活性所必需之附加的取代基或鍵結 (例如，9-酮基或9_羥基、4，5-氧四園、4-乙薩氧基、以 及2-卞醯氧基）。紫杉烷類化合物之範例爲紫杉醇和紫杉 德。 5 本紙張尺度適用中國國家標準（CNS ) Α4規格（210X297公—） (鬩讀背面之注意事項再填寫本頁) 訂 II IV nn _ 471972 A7 __—— _ _ B7五、發明説明（5 ) 經濟部中央標率局員工消費合作社印製 ”聚氧乙烯山梨醇之脂肪酸單酯”意指一種具有脫水 山梨醇核心（1,4-山梨醇環醚）之化合物，其中2,3,5-羥 基上各自有一或多個環氧乙烷單體衍生物，並且6 -羥基上 有一或多個末端爲脂肪酸單酯之環氧乙烷單體衍生物。此 化合物中，環氧乙烷單體的數目一般在10至50之間，而以 1 〇至3 〇爲佳。聚氧乙烯山梨醇之脂肪酸單酯之範例爲 ，，TWEEN 80”，亦稱爲聚氧乙烯（2 0 )山梨醇單油酸酯，其中 "(2 〇 r是指接在脫水山梨醇核心之環氧乙烷單體總數爲 20 » ”脂肪酸”意指C-16至C-22之羧酸酯，其爲完全飽 和之脂肪族或可包含一或多個碳一碳雙鍵。脂肪酸之範例 包括掠橋酸（palmitic acid)( C-16)、硬脂酸（stearic acid}( C -18>、以及油酸（okkacid)(順-9-十八烯酸> » ”聚氧乙烯山梨醇脂肪酸單酯及聚乙氧基化之篦麻油 共存，其量足以有效降低紫杉烷類化合物之毒性”意指此 單酯及篦麻油係共存，且其量足以降低紫杉烷類組成物之 毒性（尤其對稀釋後用於靜脈注射之紫杉烷類組成物而 言），此毒性乃相對於單獨使用單酯或篦麻油，取代本發 明中之單酯/篦麻油合倂使用，而達到相同程度溶解性時 之毒性。 6 本紙張尺度適用中國國家標率（CNS ) A4規格（210X 297公釐） (讀先閱讀背面之注意事項再填寫本頁) ----- 訂 • fit. 1 11 -- -i —ϋ »s-«i ·ΐί· —·— HI 1_*1 471972 A7 — B7五、發明説明（6 ) 經濟部中央標率局員工消費合作社印製 ”低分子量聚乙烧乙二醇”意指具有200至looo道爾 頓（da丨tons)平均分子量之聚乙烯乙二醇（PEG>。 ”哺乳類生體”爲其傳統上之含意，涵括例如：貓、 狗、羊、馬，以及特別是人類。 Π .紫杉烷類儲存溶液 本發明所改進之處，爲藉由一具有較低毒性的載送體 運送高劑量的紫杉烷類給癌症病人》本發明利用聚氧乙烯 山梨醇之脂肪酸單酯和聚乙氧基化之篦麻油作爲紫杉烷類 的溶解劑，可有效地提高紫杉烷類的溶解度和安定性，但 具有較低的毒性。 本發明係將具有醫藥品級純度之紫杉烷類化合物儲存 於一具有較佳溶解度和較低毒性之儲存溶液中而成，該紫 杉烷類化合物之濃度較佳爲2毫克/毫升至2 0毫克/毫 升，尤佳爲4毫克/毫升至8毫克/毫升。 該儲存溶液包括一種聚氧乙烯山梨醇脂肪酸單酯與一 種以乙醇爲基本之聚乙氧基化之篦麻油。單酯與聚乙氧基 化箆麻油共存，較諸只有聚氧乙烯山梨醇脂肪酸單酯或是 只有聚乙氧基化篦麻油的存在，其毒性可有效降低。聚乙 氧基化篦麻油之來源，只要醫藥上可接受者均可。一種適 宜的製備方法可經由BASF(Wyandotte，MD)的製程產品，其商 標爲"CREMOPHOR EL”。通常聚乙氧基化篦麻油是以 大約1 0 %到大約4 0 % ( v : v )的濃度存在，較佳的濃度爲在 7 本紙張尺度適用中國國家標準（CNS ) A4規格「210X2^7公釐） (讀先閲讀背面之注意事項再填寫本頁) 裝· 丁 責 471972 經濟部中央檬率局員工消費合作社印製 A7 B7 __Letllll__ _ ι'·π"*^*~,'*~Κπ'·-*"·" *JLWT •^•^―. --— * ΒΑ^-^··-* '' - -.-..norn^ ^ Γ ^――·. Jli* ΙΠ ~五、發明説明（7 ) 大約2 0 %到3 0 %之間。聚氧乙烯山梨醇脂肪酸單酯通常以 大約5 %到大約20 %(ν:ν)的濃度存在’較佳的濃度爲在大 約5 %到ί 5 %之間。其中較佳的聚氧乙餘山梨醇脂肪酸單 酯爲"TWEEN 80”。 聚乙氧基化篦麻油和聚氧乙烯山梨醇脂肪酸單酯共同 組成的儲存溶液，相加的濃度在大約15%到大約60% ’較 佳的濃度爲從大約25 %到45 % (v:v)。除此之外’聚乙氧 基化篦麻油和聚氧乙烯山梨醇脂肪酸單酯之混合比例爲 (篦麻油：山梨醇脂肪酸單酯’ v:v)在大約0.5到6，較 佳的比例爲大約1.3到6 ’更佳的比例爲在大約2到3 °在此 必須注意的是，聚乙氧基化篦麻油和聚氧乙嫌山梨醇脂肪 酸單酯不但可增加紫杉烷類的溶解度’同時可增加紫杉烷 類使用於治療惡性腫瘤時的抗癌效果。依據本發明的一重 要特色，利用聚乙氧基化篦麻油和聚氧乙烯山梨醇脂肪酸 單酯共存，所導致的毒性比只有聚氧乙烯山梨醇脂肪酸單 酯或是只有聚乙氧基化篦麻油的存在時所導致之毒性爲 低。 本，發明的儲存溶液，也可包含一低分子量之聚乙烯乙 二醇（P E G )，其平均分子量在大約2 0 0到1 0 0 0道爾頓 (daltoiO，較佳在約2 0 0到5 0 0道爾頓。較佳的聚乙烯乙二醇 (平均分子量在大約200到500道爾頓）在攝氏15度以上 爲液體。聚乙烯乙二醇是包含在儲存液中，以改善紫杉烷 類組成物的溶解度和安定性。在一較佳實施例中，聚乙烯 8 本紙張尺度適用中國國家標率（CNS ) A4規格(210X297公釐） (請先閱讀背面之注意事項再填寫本頁) 10. asi— mf HI a— a—·—— til-"V 、言 A « I mmtMSMW m^i —US -m 9m 經濟部中央標準局員工消費合作社印製 471972 A7 B7 一―— ____________ ______ 五、發明説明（8 ) 乙二醇之濃度爲從大約1 〇 %到6 〇 % ( v : v )之間’更佳的組 成係在約4 0 %到6 0 % ( v : v )之間。 儲存溶液中也可選擇包含一緩衝物質（buffering agent) ’此 物質可維持儲存溶液之氫離子指數（P Η値）在大約1到大約 8之間，較佳者爲大約4到大約6之間。緩衝物質的較佳選 擇爲配藥上可接受的酸，更佳的選擇爲一羧酸’例如檸檬 酸、醋酸、順丁烯二酸、丁二酸、乳酸、抗壞血酸、銨基 戊二酸，或是天門冬銨酸（aspartic add)。其中尤以無水檸檬 酸爲佳。緩衝物質可存在的濃度大約在2到大約200mM之 間，一般是在大約5到大約2 0 0 m Μ之間。 至於該儲存溶液之其它部分則包含乙醇，溶液中以不 包含水爲佳。 本發明的儲存溶液可以任何合適的方法溶解紫杉烷類 化合物，包括利用超音波和加熱。後文中將舉例說明之實 例1中提供了一種配製本發明溶液的方法。此溶液可在室 溫下儲存，但較佳儲存於4 °C或更低的溫度下。溶液最好 經過一過瀘膜（filter membrane)以除去小的顆粒，例如大小爲 0.22μηι的過濾膜。該溶液亦可藉由通入氮氣以移去氧 氣。 本發明中儲存液之安定性，將於實例2與實例3中加以 說明。在實例2之相關說明中，按照本發明所配製的兩種 儲存溶液，放置在一高壓滅菌器內，在加壓之250 °C溫度 條件下加熱2 0分鐘。樣品經由乙腈稀釋，以高效能液相層 9 本紙張尺度適用中國國家標準（CNS ) A4规格（21〇X297公釐） .1 I 1 I ! ................ li I . (閱讀背面之注意事項再填寫本頁)In n PI! 471972 A7 __ —— —________ B7 V. Description of the invention (2) The surfactant cooperative agent printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs, such as a fatty acid ester of polyoxyethylene sorbitol (also known as " TWEEN 80 ", also known as polySOrbate), polyethoxylated ramie oil (aka" CreniophorEL "), soybean oil or triacetate" but these dosage forms do not experience low solubility (especially in physiological salt When diluting in water for intravenous injection), you are experiencing high toxicity problems. High toxicity problems are caused by oils or surfactants. Especially when the concentration of "TWEEN 80" is high enough to dissolve high concentrations of paclitaxel In clinical practice, it can cause pleural effusions and edema, and "Cremophor EL" can cause severe and even fatal hypersensitivity reactions (RM Straubinger, " Taxol: Science and Applications "(M Stiffness Editor}, CRC Press, New York, NY, pp. 241 and 244, 199 S). Therefore, there is an urgent need to improve the formulation of paclitaxel to maintain its longevity. Under the premise of storage stability, it can have low toxicity. The object of the present invention is to provide a taxane composition, which is suitable for intravenous pharmaceutical use due to its high stability and low toxicity, and can be used for Preventing the continued growth of biological cancer ^ One aspect of the present invention relates to a taxane composition, which is obtained by storing a taxane compound in a storage solution with better solubility and lower toxicity, wherein The storage solution includes: (a) a fatty acid monoester of polyoxyethylene sorbitol; (b) polyethoxylated ramie oil; and (c >ethanol; in this solution, the monoester and polyethoxylated The coexistence of ramie oil, the paper size of this paper applies the Chinese National Standard (CNS) A4 specification (210X297 mm) (please read the precautions on the back of the results before filling out this page): # ............ ............. 0 471972 A7 B7, ^ ........... .......... ..... LUI. 11 «» " —— r —I --- 1 — ........ ...........................-..........._ l I ΙΙ. · »· _ · .Ι 'τ ._ .. Ι..1, .ΙΙ.ΙΙΙΙ.ΊΙ.! ..... ....... u, „^ _. V. Description of the invention (3) The amount printed by the employee consumer cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs is sufficient to effectively reduce taxanes The composition is toxic, and the p Η 値 of the storage solution is preferably between about 1 and 8. Wherein, the fatty acid monoester of the polyoxyethylene sorbitol is preferably polyoxyethylene (20) sorbitol monooleate; and the storage solution preferably further includes a low molecular weight polyethylene glycol, such as FEG 30 0 » In a preferred embodiment, the taxane composition comprises 4 mg / ml to 8 mg / ml of a taxane compound, 20 to 30% (ν / ν) polyethoxylated ramie oil, s To 1S% (v / v) polyoxyethylene (20) sorbitol monooleate, 15 to 3% (v / v > ethanol and 40 to 60% (v / v & gt) low molecular weight polyethylene Among them, the storage solution preferably further comprises-· acid as a buffering agent, so that p 其中 therein is maintained between about 4 and 6. The acid may be anhydrous citric acid. Among them, the purple The taxane compound may be paclitaxel or decetaxel. Another aspect of the present invention relates to a pharmaceutical taxane composition suitable for intravenous injection. The taxane composition is a taxane Compounds are stored in a storage solution with better solubility and lower toxicity, and then diluted with a diluent suitable for intravenous injection. The storage solution includes: (a > a fatty acid monoester of polyoxyethylene sorbitol; (b > polyethoxylated ramie oil; and (c) ethanol; in this solution, the monoester and polyethoxylated The coexistence of ramie oil is sufficient to effectively reduce the toxicity of the taxane composition, and the pH of the storage solution is preferably between about 1 and 8. Among them, the diluent is physiological saline. 4 papers The scale is applicable to China National Standard (CNS) A4 (210X297 mm) (read the precautions on the back before filling this page) if installed --- 88 »: '·-1Β 8ϋ order tor • m -ml f« i 8 * 1 ί_1 I,:. SI > 471972 A7 _ ——— B7 V. Description of the invention (4) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs Another aspect of the present invention relates to the use of taxanes A method for treating mammalian cancers with the composition, comprising the step of injecting a pharmaceutical taxane composition suitable for intravenous injection into a living body in a pharmaceutically acceptable amount 'wherein the taxane composition is Taxanes are stored in a form with better solubility and lower toxicity It is diluted with a suitable diluent suitable for intravenous injection, and the storage solution includes: (3 > fatty acid monoester of polyoxyethylene sorbitol; (b) polyethoxylated ramie oil ; And (C >ethanol; in this solution, the monoester and polyethoxylated ramie oil coexist in an amount sufficient to effectively reduce the toxicity of the taxane composition, and the P of the storage solution should be within about It is preferably between 1 and 8. Other objects and characteristics of the present invention will be described in detail below. Detailed description of the present invention I. Definitions of the terms used below have the following meanings: "Taxanes" means anything that can inhibit micro A depoymerized compound having a paclitaxel 6-8-6 fused ring skeleton 'the skeleton includes additional substituents or bonds necessary for paclitaxel activity (e.g., 9-keto or 9-hydroxy, 4,5-oxotetracycline, 4-ethexyloxy, and 2-fluorenyloxy). Examples of taxanes are paclitaxel and taxol. 5 This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 male —) (read the notes on the reverse side and fill out this page) Order II IV nn _ 471972 A7 __—— _ _ B7 V. Description of the invention (5) The "Fatty Acid Monoester of Polyoxyethylene Sorbitol" printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs means a compound with a sorbitan core (1,4-sorbitol cyclic ether), of which 2,3,5- Each of the hydroxyl groups has one or more ethylene oxide monomer derivatives, and the 6-hydroxyl group has one or more ethylene oxide monomer derivatives terminated with a fatty acid monoester. In this compound, the number of ethylene oxide monomers is generally between 10 and 50, and preferably between 10 and 30. An example of a fatty acid monoester of polyoxyethylene sorbitol is, TWEEN 80 ", also known as polyoxyethylene (20) sorbitol monooleate, where " (2 〇 means refers to the sorbitan core The total number of ethylene oxide monomers is 20 »" Fatty acid "means C-16 to C-22 carboxylic acid esters, which are fully saturated aliphatic or may contain one or more carbon-carbon double bonds. Examples include palmitic acid (C-16), stearic acid (C-18), and okkacid (cis-9-octadecenoic acid) »" " The coexistence of sorbitol fatty acid monoester and polyethoxylated ramie oil in an amount sufficient to effectively reduce the toxicity of taxane compounds "means that the monoester and ramie oil coexist in an amount sufficient to reduce the composition of taxanes Toxicity (especially for the taxane composition diluted for intravenous injection), this toxicity is relative to the monoester or ramie oil used instead of the monoester / ramie oil combination in the present invention, The toxicity when reaching the same degree of solubility. 6 This paper size applies Chinese National Standard (CNS) A4 regulations (210X 297mm) (Read the precautions on the back before filling this page) ----- Order • fit. 1 11--i —ϋ »s-« i · ΐί · — · — HI 1_ * 1 471972 A7 — B7 V. Description of the invention (6) “Low molecular weight polyethylene glycol” printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs means that it has an average molecular weight of 200 to looo daltons. Polyethylene glycol (PEG). "Mammal" is its traditional meaning, including, for example, cats, dogs, sheep, horses, and especially humans. Π. Taxane storage solutions improved by the present invention In order to deliver high doses of taxanes to cancer patients through a carrier with lower toxicity, the present invention uses polyoxyethylene sorbitol fatty acid monoester and polyethoxylated ramie oil as purple Taxane dissolving agents can effectively improve the solubility and stability of taxanes, but have low toxicity. The present invention is to store taxane compounds with pharmaceutical grade purity in a better solubility And less toxic storage solutions. The concentration is preferably 2 mg / ml to 20 mg / ml, particularly preferably 4 mg / ml to 8 mg / ml. The storage solution includes a polyoxyethylene sorbitol fatty acid monoester and an ethanol-based polyethylene. Oxylated ramie oil. The co-existence of monoester and polyethoxylated ramie oil, its toxicity can be effectively reduced compared to the presence of only polyoxyethylene sorbitol fatty acid monoester or only polyethoxylated ramie oil. The source of ethoxylated ramie oil is acceptable as long as it is medically acceptable. A suitable preparation method can be manufactured by BASF (Wyandotte, MD). Its trademark is "CREMOPHOR EL". Polyethoxylated ramie oil is usually present at a concentration of about 10% to about 40% (v: v). The preferred concentration is 7 paper sizes, which is applicable to the Chinese National Standard (CNS) A4 specification "210X2 ^ 7mm) (Read the precautions on the back before filling in this page) Packing · Ding 471972 Printed by the Consumers' Cooperative of the Central Bureau of the Ministry of Economic Affairs A7 B7 __Letllll__ _ ι '· π " * ^ * ~,' * ~ Κπ '·-* " · " * JLWT • ^ • ^ ―. --- * ΒΑ ^-^ ··-*' '--.- .. norn ^ ^ Γ ^ —— ·. Jli * ΙΠ ~ V. Description of the invention (7) About 20% to 30%. Polyoxyethylene sorbitol fatty acid monoester usually exists at a concentration of about 5% to about 20% (ν: ν). The preferred concentration is Between about 5% and 5%. The preferred polyoxyethylene sorbitan fatty acid monoester is " TWEEN 80 ". The storage solution composed of polyethoxylated ramie oil and polyoxyethylene sorbitol fatty acid monoester is added at a concentration of about 15% to about 60%. The preferred concentration is from about 25% to 45% (v: v). In addition, the mixing ratio of 'polyethoxylated ramie oil and polyoxyethylene sorbitol fatty acid monoester is (ramie oil: sorbitol fatty acid monoester' v: v) is about 0.5 to 6, and the preferred ratio is About 1.3 to 6 'The better ratio is at about 2 to 3 °. It must be noted here that polyethoxylated ramie oil and polyoxyethylene sorbitan fatty acid monoester can not only increase the solubility of taxanes' At the same time, it can increase the anti-cancer effect of taxanes in the treatment of malignant tumors. According to an important feature of the present invention, the coexistence of polyethoxylated ramie oil and polyoxyethylene sorbitol fatty acid monoester results in a toxicity ratio that is only polyoxyethylene sorbitol fatty acid monoester or only polyethoxylated rhenium The toxicity caused by sesame oil is low. The storage solution of the present invention may also contain a low molecular weight polyethylene glycol (PEG), the average molecular weight of which is about 200 to 100 Dalton (daltoiO, preferably about 200 to 500 Daltons. The preferred polyethylene glycol (average molecular weight between about 200 and 500 Daltons) is liquid above 15 degrees Celsius. Polyethylene glycol is contained in a storage solution to improve violet Solubility and stability of taxane composition. In a preferred embodiment, the paper size of polyethylene 8 is applicable to China National Standard (CNS) A4 specification (210X297 mm) (Please read the notes on the back before filling (This page) 10. asi— mf HI a— a— · —— til- " V, say A «I mmtMSMW m ^ i —US -m 9m — ____________ ______ 5. Description of the invention (8) The concentration of ethylene glycol is from about 10% to 60% (v: v). The better composition is about 40% to 60% (v: v). A buffering agent can also be included in the storage solution. 'This substance can maintain the storage solution. The hydrogen ion index (P Η 値) is between about 1 and about 8, preferably between about 4 and about 6. The preferred choice of buffer material is a pharmaceutical acceptable acid, and the more preferred choice is monocarboxylic acid. Acids such as citric acid, acetic acid, maleic acid, succinic acid, lactic acid, ascorbic acid, ammonium glutaric acid, or aspartic add. Among them, anhydrous citric acid is preferred. Buffer substances Concentrations that can exist are between about 2 and about 200 mM, typically between about 5 and about 200 mM. As for the other parts of the storage solution, they contain ethanol, and the solution preferably contains no water. The present invention The storage solution can dissolve taxane compounds by any suitable method, including the use of ultrasound and heating. A method for preparing the solution of the present invention is provided in Example 1 which will be exemplified later. This solution can be stored at room temperature. However, it is best stored at 4 ° C or lower. The solution is best passed through a filter membrane to remove small particles, such as a 0.22 μm filter membrane. The solution can also be passed through Nitrogen shift Oxygen. The stability of the storage solution in the present invention will be described in Examples 2 and 3. In the related description of Example 2, two storage solutions prepared according to the present invention are placed in an autoclave, Heated under pressure at 250 ° C for 20 minutes. The sample was diluted with acetonitrile to form a high-performance liquid layer. 9 This paper is sized to the Chinese National Standard (CNS) A4 (21 × 297 mm). 1 I 1 I ! ...... li I. (Read the notes on the back and fill in this page)
、tT 471972 A7 B7 經濟部中央梂準局員工消費合作社印製 五、發明説明（ 析儀（HPLC)分析，沒有偵測到任何紫杉醇降解（degradation) 的情況發生。 在實例3所描述之硏究中，樣品溶液在培養箱內，以 3 7 °C培養1 2週，定時取出少量水溶液，利用高效能液相層 析儀分析紫杉醇的降解情形。受測試的樣品溶液包含實例 1中的劑型1、劑型2和一包含紫杉醇之聚乙氧基化篦麻油 和乙醇以1 : 1比例混合之溶液（劑型3 )。由實例3之結果 可看出，按照本發明所配製的紫杉醇溶液至少和劑型3 — 樣安定，在1 2週內之降解比例少於2 %。 依照本發明之另一重要特色爲，本發明之紫杉烷類組 成物經由稀釋後，和藥物靜脈注射標準溶液是相容的。在 實例四所說明的硏究中，將實例1中之劑型以生理食鹽水 (0.9%氯化鈉水溶液）稀釋之，稀釋量爲5倍、10倍、25 倍和5 0倍，在1小時、2小時、4小時、8小時、2 4小時和 4 8小時後，檢查溶液是否有沉澱或是混濁。結果發現劑型 1在4 8小時後仍保持澄淸。由此結果顯示，依本發明所配 製的儲存溶液適合靜脈注射使用。 實例5所說明之硏究，爲單獨之儲存載送體（無紫杉 醇之儲存溶液），和一由聚乙氧基化篦麻油和乙醇以1:1 比例混合之溶液（劑型3 )比較其相對毒性在一實驗 中，2或3隻老鼠爲一組，注射一劑被稀釋的測試劑型，觀 察老鼠在21天內因注射所引起之無耐受性（intolerance)。無耐 受性的跡象包括以下所述（1)明顯的體重減輕（>2〇%)，（2> 豎毛（pi丨©erection)，（ 3 }長期虛脫（prolonged prostration)和（4 )死 10 本紙張尺度適用中國國家標率（CNS ) A4规格（2107^7公釐） (請先閱讀背面之注意事項再填寫本頁) S3S8· ·1· mi-_·— am— In· V .· 缘 、\呑 471972 A7 B7五、發明説明（10 ) 經濟部中央標準局員工消費合作社印製 亡。將不會產生無耐受性之最高注射劑量（MTD，最大容 忍劑量）記錄下來。由實例SA的結果看出，依本發明所配 製的劑型，其最大容忍劑量是單獨使用聚乙氧基化箆麻油 而不含山梨醇單油酸酯者之兩倍。當同樣的劑型係以小劑 量/體積在5天內每6小時注射一次時，亦可得到和上述者 相近之結果。同樣的，依本發明所配製的劑型，其最大容 忍劑量是單獨使用聚乙氧基化篦麻油者之兩倍。這些結果 顯示本發明的載送體，和單獨使用聚乙氧基化篦麻油而不 含山梨醇單油酸酯者相比較，具有較低的故有毒性（inherent toxicity)。如此便可容許較大之紫杉烷類注射量；或是在注 射等量紫杉烷類之條件下，其毒性副作用較輕。由此可 知，習用紫杉醇類劑型具有載送體本身具有毒性之副作 用，以及紫杉烷類組成物注射劑量受限制的缺點》而本發 明則在以上兩方面明顯優於習用的劑型。 皿·治療方法（Treatment Method} 另一方面，本發明關於治療哺乳類動物癌症之方法。 在此方法中，將一適於靜脈注射之醫藥用紫杉烷類組成 物，以醫藥上可接受之量注射入生體中，以粗止生體癌症 之繼續成長。其中該紫杉烷類組成物係將紫杉烷類化合物 儲存於前述儲存溶液中，再以一適於靜脈注射之稀釋劑稀 釋而成。 11 本紙張尺度適用中國國家標準（CNS ) A4規袼（2i〇x297公釐） (請先閱讀背面之注意Ϋ項再填寫本頁), TT 471972 A7 B7 Printed by the Consumer Cooperatives of the Central Bureau of Associate Bureau of the Ministry of Economic Affairs 5. Analysis of the invention (HPLC) analysis, did not detect any degradation of paclitaxel. The investigation described in Example 3 In the incubator, the sample solution was cultured at 37 ° C for 12 weeks. A small amount of aqueous solution was periodically taken out, and the degradation of paclitaxel was analyzed by high-performance liquid chromatography. The sample solution tested contained dosage form 1 in Example 1. Formulation 2 and a solution of paclitaxel-containing polyethoxylated ramie oil and ethanol in a 1: 1 ratio (Formulation 3). From the results of Example 3, it can be seen that the paclitaxel solution prepared according to the present invention is at least the same as the dosage form. 3 — Stable, with a degradation rate of less than 2% within 12 weeks. According to another important feature of the present invention, the taxane composition of the present invention is compatible with a standard drug intravenous solution after dilution. In the research described in Example 4, the dosage form in Example 1 was diluted with physiological saline (0.9% sodium chloride aqueous solution), and the dilutions were 5 times, 10 times, 25 times, and 50 times. 1 hour, 2 After 4 hours, 8 hours, 24 hours, and 48 hours, the solution was checked for precipitation or turbidity. As a result, it was found that dosage form 1 remained clear after 48 hours. The results showed that the formulation according to the present invention The storage solution is suitable for intravenous injection. The investigation described in Example 5 is a separate storage carrier (a paclitaxel-free storage solution) and a mixture of polyethoxylated ramie oil and ethanol in a 1: 1 ratio. The relative toxicity of the solution (formulation 3). In one experiment, 2 or 3 mice were used as a group, and one dose of the diluted test formulation was injected. The mice were observed for intolerance caused by the injection within 21 days. Signs of intolerance include the following (1) significant weight loss (> 20%), (2) vertical hair (pi 丨 © erection), (3) long-term prostration, and (4) Dead 10 This paper size applies to China National Standards (CNS) A4 specification (2107 ^ 7 mm) (Please read the precautions on the back before filling this page) S3S8 ·· 1 · mi-_ · — am— In · V . · Yuan, \ 呑 471972 A7 B7 V. Description of Invention (10) Central Ministry of Economic Affairs Printed by the Consumer Bureau of Standards Bureau. The highest injection dose (MTD, maximum tolerated dose) that does not produce intolerance is recorded. From the results of Example SA, it can be seen that the dosage form formulated according to the present invention has the maximum tolerance The dose is twice that of polyethoxylated ramie oil without sorbitol monooleate. When the same dosage form is injected every 6 hours in a small dose / volume over 5 days, and The above results are similar. Similarly, the maximum tolerated dose of the dosage form formulated according to the present invention is twice that of the polyethoxylated ramie oil alone. These results show that the carrier of the present invention has lower inherent toxicity than the case where polyethoxylated ramie oil is used alone without containing sorbitol monooleate. This allows a larger injection volume of taxanes, or it has less toxic and side effects when the same amount of taxanes are injected. From this, it can be known that the conventional taxol-based dosage form has the side effect of the carrier itself being toxic, and the disadvantages of the limited dosage of the taxane composition. The present invention is significantly superior to the conventional dosage form in the above two aspects. [Treatment Method] On the other hand, the present invention relates to a method for treating mammalian cancer. In this method, a pharmaceutical taxane composition suitable for intravenous injection is used in a pharmaceutically acceptable amount. It is injected into the living body to stop the growth of cancer in the crude. The taxane composition is to store the taxane compound in the aforementioned storage solution, and then dilute it with a diluent suitable for intravenous injection. 11 This paper size is subject to Chinese National Standard (CNS) A4 (2i0x297 mm) (Please read the note on the back before filling this page)
471972 A7 B7 經濟部中央標準局員工消费合作社印製 五、發明説明（11 ) 藉由稀釋後紫杉烷類組成物之注射，可有效阻止或消 滅癌症的成長，以治療任何的癌症狀況，包括如卵巢癌、 乳癌、膀胱癌、肺癌、黑色素癌和淋巴瘤癌等。 在本發明中使用的稀釋劑，可爲任何適用於靜脈注射 者。一般而言，此稀釋溶液包括氯化鈉，以建立一選擇性 的生理滲透克分子濃度（physio丨ogica丨osmolarity)，例如0.9 % (重量/體積）氯化鈉溶液。此稀釋溶液可另外加上合適 的補充劑’例如葡萄糖和/或一抗生素，例如盤尼西林或 四環素。此溶液較佳以非塑膠容器承裝，以免塑化劑浸瀝 (leaching)入溶液中。此稀釋紫杉烷類溶液以一選擇的速度注 射，直到所需要的藥量完全注入。此劑型是以定期的注射 方式注射，直到獲得緩解（remission)的狀態，或是癌症腫瘤停 止成長（proliferation)。在癌症手術後一段時間，顯示癌瘤已成 功的割除時，亦可注射此劑型以阻止癌症的再生》利用紫 杉醇和紫杉醇衍生物來治療癌症病人，記錄在S. G. Arbuck and B. A. Biaylock，”Taxol: Science and Applications”（M. Stiffness編輯）， C R C 刊印，New York，NY，第 379-415頁，1 9 9 5 )中。 本發明之醫藥用紫杉烷類組成物之另一優點爲其可與 其它適用於癌症病人之抗癌藥物合併使用。例如，其可與 西伯拉丁（ cisplatin)，亞達去散（edatrexate)，必塞寧亞諷（L-buthionine sulfoxide}，三唾块靈（triazofurin)，硝酸鎵（gaHium nitrate),多索如比辛 (doxorubicin),依托普塞（etoposide)或環磷醯胺（cyclophosphamide)—起 使用，亦可和放射線治療合倂使用。不僅如此，先前的討 論中，描述本發明載送體在紫杉醇利用上之優點，然而， 12 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) "5---!二-=s_ ---Ms-—--· ......----- -sin- . - UK--SJ-.1_—I —I— ii . p 471972 A7 B7 _ u , τ„·,...... ........... τ - I111'亂 L_ ------------—— — 一 _ 五、發明説明（I2) 本發明載送體並已被考慮到應用於其它非極性紫杉醇/紫 杉烷類衍生物（例如d 〇 c e t a X e丨），無論其爲合成物或天 然產物。 以下的實例係用以舉例說明本發明，而非意欲限制任 何本發明之利用。 實例1 紫杉醇劑型 依照以下比例配製了兩種劑型，以作爲以·F硏究之 用。 (請先聞讀背面之注意事項再填寫本頁} .裝. 劑型1 劑型2 PEG 2 0 ml 2 5 ml 絕對乙醇 10 ml 10 ml 無水檸檬酸 10 0 mg 10 0 mg Cremophore EL 15 ml 10 ml Tween 80 5 ml 5 ml 太平洋紫杉醇 3 0 0 mg 3 0 0 mg 最終體積 5 0 ml 50 ml -訂--- 經濟部中央標準局員工消費合作社印製 爲了製備以上的劑型，檸檬酸和乙醇利用一高速混合 器或一攪拌匙混合，直到完全溶解爲止。必要的話，混合 液可加熱至S 0 °c或利用超音波使其完全溶解。將聚氧乙烯 山梨醇之脂肪酸單酯和聚乙氧基化之篦麻油加入混合液 中，以高速攪拌器攪拌30分鐘。將紫杉醇加入混合液中’ 13 本紙張尺度適用中國國家檩準（CNS ) A4規格（2ί0Χ29ί7公釐） 471972 經濟部中央標準局員工消費合作杜印製 A7 __^ B7 五、發明説明（13) 攪拌直到紫杉醇完全溶解。此溶液以乾燥氮氣通過後’經 過〇·22微米過濾器（"MILLIPACK” 200>。以上兩劑型 所得之紫杉醇最終濃度爲6毫克/毫升。 實例2溫度的安定性 劑型1和劑型2各取200微升，置入一 2毫升琥珀小瓶 中，以乾燥氮氣通過後，使用包有聚四氟乙烯的橡皮塞 住，然後利用鋁箔密封。此密封的小瓶置入高壓滅菌器’ 在壓力下於華式2 5 G度加熱2 Q分鐘。經過高壓滅菌的樣 品，以乙腈稀釋（1 ·· 2 0 )，利用Waters C8 Novapak管柱，以高 效能液相層析儀分析（8mm I.D. X 10cm，緩衝溶液A: 乙腈/水/三氟乙酸= 20/80/0.1 ;緩衝溶液B:乙腈/水 /三氟乙酸= 80/20/0.1; 45 % B溶液之恆常梯度； 230nm下偵測）。高效能液相層析儀分析結果顯示，紫杉 醇沒有降解的跡象。 實例3 劑型長時期安定性之比較 劑型1和劑型2各取2 00微升和劑型3 (包括6毫克/毫 升紫杉醇，在1/1的聚乙氧基化篦麻油和乙醇混合液）置 入一 2毫升琥珀小瓶中，以乾燥氮氣通過後密封，置入一 37°C的加熱箱。50微升的溶液分別在1，3,6和12週取 出，以HPLC級乙腈稀釋（1:20)，利用高效能液相層析儀 分析。結果詳列如下： 14 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） 請先閱讀背面之注意事項再填寫本頁) « 111 HB^l Ilf nn * .11 1=11=1-1 · 訂 471972 A7 B7 五、發明説明（14 ：) 時間（週） 劑型 紫杉醇殘餘百分比 # 1 #2 #3 1 1 0 0 loo 10 0 6 9 8.8 9 8.8 98.7 12 98.7 9 8.8 9 7.8 實例4 紫杉醇劑型的安定性 依照劑型1 和劑型2所配製的紫杉醇組成物 ’以 0.9 % 食鹽水稀釋5、 iO ' 25和 50倍 ’得到紫杉醇的濃度分別爲 ....... ! = -- I .......... ......... ........ .......... * (讀先閱讀背面之注意Ϋ項再填寫本頁) 1.2、0.6、0.24和〇·12毫克/毫升。這些溶液分別在1 ' 2、4、8、24和48小時，查看是否有沉澱或是混濁度。 所有劑型1的稀釋溶液，在4 8小時內都保持澄淸，沒 有沉澱或混濁的跡象。所有劑型2的稀釋溶液，在24小時 內都保持澄淸，但在4 8小時內都有沉澱或混濁的跡象。以 稀釋5倍的劑型2，有最多的沉澱。 實例S 紫杉醇劑型毒性的比較 A .未稀釋樣品毒性的比較 紫杉醇劑型1、2和3未稀釋樣品毒性的比較’以 B a丨b / c老鼠測試急性毒性試驗。樣品以固定範圍體積’靜 脈注射到以2或3隻爲一組，18至20克重的老鼠。在注射 後21天內觀察老鼠無耐受性的程度。無耐受性的跡象包括 以下所述（1)明顯的體重減輕（>20% >，（2>豎毛 15 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） -訂--- 經濟部中央標準局員工消費合作社印製 471972 A7 ______ B7 五、發明説明（is) (piloerection}，（ 3 )長期虛脫（pro丨⑽ged prostration)和（4 )死亡。結 果列在下表。最大容忍劑量（Μ T D )以毫升/公斤爲單位。 型 最大容忍劑量（毫升/公斤） 老鼠數目 # 1 5.0 2 #2 5.0 3 #3 <2.5 3 Β.長期注射的毒性試驗 紫杉醇劑型1、2和3，以生理食鹽水1 : 1稀釋樣品。以 靜脈注射注入18至20克之Balb/c老鼠，一天注射4次，共 注射五天。在注射後2 1天內觀察老鼠無耐受性的程度。最 大容忍累積劑量列於下表： (請先閱讀背面之注意事項再填寫本頁) ϋ ml I— nn 111 —I— V .* 03. 、言 型 最大容忍劑量（毫升/公斤） 老鼠數目 # 1 10 5 # 2 10 5 #3, 5.0 5 經濟部中央標準局員工消費合作社印製 本發明得藉熟習此技藝之人士任施匠思而爲諸般修 飾，然皆不脫如附申請範圍所欲保護者。471972 A7 B7 Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 5. Description of the Invention (11) Injection of diluted taxane composition can effectively prevent or eliminate the growth of cancer to treat any cancer condition, including Such as ovarian cancer, breast cancer, bladder cancer, lung cancer, melanoma cancer and lymphoma cancer. The diluent used in the present invention may be any suitable for intravenous injection. Generally, the diluted solution includes sodium chloride to establish a selective physiological osmolarity, such as a 0.9% (w / v) sodium chloride solution. This diluted solution may be additionally supplemented with a suitable supplement ' such as glucose and / or an antibiotic such as penicillin or tetracycline. This solution is preferably contained in a non-plastic container to prevent the plasticizer from leaching into the solution. This diluted taxane solution is injected at a selected rate until the required dose is fully injected. This dosage form is injected by regular injections until remission is achieved, or cancer tumors stop growing (proliferation). After a period of cancer surgery, showing that the cancer has been successfully removed, this dosage form can also be injected to prevent cancer from regenerating. "Using paclitaxel and paclitaxel derivatives to treat cancer patients, recorded in SG Arbuck and BA Biaylock," Taxol: Science and Applications "(edited by M. Stiffness), CRC Press, New York, NY, pp. 379-415, 195 5). Another advantage of the pharmaceutical taxane composition of the present invention is that it can be used in combination with other anticancer drugs suitable for cancer patients. For example, it can be combined with cisplatin, edatrexate, L-buthionine sulfoxide, triazofurin, gaHium nitrate, dosorubicin Doxorubicin, etoposide or cyclophosphamide can be used in combination with radiation therapy. Moreover, in the previous discussion, the carrier of the present invention was described as being used in paclitaxel. Advantages, however, 12 paper sizes are applicable to China National Standard (CNS) A4 specifications (210X297 mm) (Please read the precautions on the back before filling this page) " 5 ---! 二-= s_ --- Ms ----- · ......----- -sin-.-UK--SJ-.1_—I —I— ii. P 471972 A7 B7 _ u, τ „·, ... ... ........... τ-I111 'chaos L_ ------------—— — _ 5. Description of the invention (I2) The carrier of the invention and It has been considered for application to other non-polar paclitaxel / taxane derivatives (such as doceta X e 丨), regardless of whether they are synthetic or natural products. The following examples are intended to illustrate the present invention and are not intended to be Restricted What is the use of the present invention. Example 1 Two formulations of paclitaxel were prepared according to the following ratios for research purposes. (Please read the precautions on the back before filling out this page}. Packing. Formulation 1 Formulation 2 PEG 2 0 ml 2 5 ml absolute ethanol 10 ml 10 ml anhydrous citric acid 10 0 mg 10 0 mg Cremophore EL 15 ml 10 ml Tween 80 5 ml 5 ml paclitaxel 3 0 0 mg 3 0 0 mg final volume 50 ml 50 ml -Order --- Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs To prepare the above dosage forms, citric acid and ethanol are mixed with a high-speed mixer or a stirring spoon until completely dissolved. If necessary, the mixture can be heated to S 0 ° c or use ultrasound to completely dissolve it. Add the fatty acid monoester of polyoxyethylene sorbitol and polyethoxylated ramie oil to the mixture and stir with a high-speed stirrer for 30 minutes. Add paclitaxel to the mixture Medium '13 This paper size is applicable to China National Standards (CNS) A4 (2ί0 × 29ί7mm) 471972 Employees' cooperation of the Central Standards Bureau of the Ministry of Economic Affairs Du printed A7 __ ^ B7 V. Description of the invention (13) Stirring Until paclitaxel is completely dissolved. This solution was passed through dry nitrogen through a 22-micron filter (" MILLIPACK "200 >. The final concentration of the paclitaxel obtained in the above two dosage forms was 6 mg / ml. Example 2 Temperature stability formulation 1 and dosage form 2 were each taken 200 microliters, placed in a 2 ml amber vial, passed with dry nitrogen, sealed with a polytetrafluoroethylene rubber stopper, and sealed with aluminum foil. This sealed vial was placed in an autoclave under pressure. Chinese-style heating at 2 5 G for 2 Q minutes. The autoclaved samples were diluted with acetonitrile (1 ·· 20) and analyzed using a Waters C8 Novapak column with high performance liquid chromatography (8mm ID X 10cm, Buffer solution A: acetonitrile / water / trifluoroacetic acid = 20/80 / 0.1; buffer solution B: acetonitrile / water / trifluoroacetic acid = 80/20 / 0.1; constant gradient of 45% B solution; detection at 230nm) HPLC analysis showed no signs of paclitaxel degradation. Example 3 Comparison of Long-Term Stability of Dosage Forms Take Form 200 and 2 each of Form 1 and Form 3 (including 6 mg / ml paclitaxel, in 1/1 polyethoxylated ramie oil mixed with ethanol Mixture) into a 2 ml amber vial, sealed with dry nitrogen, and placed in a 37 ° C heating box. 50 microliters of solution were taken out at 1, 3, 6 and 12 weeks, respectively, and HPLC grade acetonitrile Diluted (1:20) and analyzed by high performance liquid chromatography. The results are detailed as follows: 14 This paper size applies to China National Standard (CNS) A4 (210X297 mm) Please read the precautions on the back before filling in this Page) «111 HB ^ l Ilf nn * .11 1 = 11 = 1-1 · Order 471972 A7 B7 V. Description of the invention (14 :) Time (week) Residual percentage of paclitaxel in dosage form # 1 # 2 # 3 1 1 0 0 loo 10 0 6 9 8.8 9 8.8 98.7 12 98.7 9 8.8 9 7.8 Example 4 Stability of paclitaxel dosage form Paclitaxel composition formulated according to dosage form 1 and dosage form 'diluted with 0.9% saline 5, iO' 25 and 50 times' The concentration of paclitaxel is .........! =-I .......... ............... ..... * (read the note on the back first and then fill out this page) 1.2, 0.6, 0.24 and 〇12 mg / ml. These solutions are at 1 '2, 4, 8, 24 and 48 hours, respectively. See if there is precipitation Is the turbidity. The solution was diluted 1 All formulations, Cheng Qing remains within 48 hours, and no signs of cloudiness or precipitation. The diluted solutions of all dosage forms 2 remained clear for 24 hours, but showed signs of precipitation or turbidity within 48 hours. With dosage form 2 diluted 5 times, there was the most precipitation. Example S Comparison of Toxicity of Paclitaxel A. Comparison of Toxicity of Undiluted Samples Comparison of Toxicity of Paclitaxel 1, 2, and 3 Undiluted Samples' Acute toxicity tests were performed with Baab / c mice. Samples were injected intravenously in a fixed range volume ' to mice in groups of 2 or 3, weighing 18 to 20 grams. The degree of intolerance of the mice was observed within 21 days after the injection. Signs of intolerance include the following: (1) Significant weight loss (> 20% >, (2 > vertical hair 15) This paper size applies Chinese National Standard (CNS) A4 specifications (210X297 mm)-order --- Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 471972 A7 ______ B7 V. Description of the invention (is) (piloerection), (3) prolonged prostration and (4) death. The results are listed in the table below. The maximum tolerated dose (M TD) is expressed in ml / kg. The maximum tolerated dose (ml / kg) is the number of rats # 1 5.0 2 # 2 5.0 3 # 3 < 2.5 3 Β. Toxicity test for long-term injection Paclitaxel dosage form 1, 2 and 3, dilute the sample 1: 1 with saline. Intravenous injection of 18 to 20 g of Balb / c mice, 4 times a day for a total of 5 days. Observe the mice for tolerance within 21 days after injection The maximum tolerated cumulative dose is listed in the following table: (Please read the precautions on the back before filling this page) ϋ ml I— nn 111 —I— V. * 03. 、 Typical maximum tolerated dose (ml / kg) Number of rats # 1 10 5 # 2 10 5 # 3, 5.0 5 Warp Central Bureau of Standards portion printed consumer cooperative employees obtained by the present invention, those skilled in the art of this application to any thinking Carpenter and repair of all sorts of ornaments, however neither application range off as desired attachment protector.
Mr -I tit ϋϋ I ml , 16 本紙張尺度適用中國國家標準（CNS ) 21〇χ297公釐）Mr -I tit ϋϋ I ml, 16 sizes of paper are applicable to Chinese National Standard (CNS) 21〇 × 297mm