1270379 玖、發明說明： [發明所屬之技術領域] 本發明關於製造脂質體多索如必辛（doxorubicin)藥學組成 物之方法，及其用於治療具有胰臟癌的用途。 [先前技術] 在2003年，在美國，約30,000病患死於胰臟癌，以及診斷 出約30,700名新病例，使得胰臟癌成為與癌症有關之死亡率的十 大最常見原因之一。對許多病患而言，手術、放射線治療，以及 化學療法為延續生命及/或減輕症狀的治療選擇，但很少治癒 (Cancer Facts and Figures，American Cancer Society 2003 )。 與非脂質體藥物相較，脂質體藥物在靜脈注射至動物及人類 後’可相當成功地到達原發性腫瘤及其轉移，以及顯示出較強的 治療效果，以及較輕微的副作用。最廣為發展之脂質體藥物之一 為脂質體多索如必辛（doxorubicin)。在台灣，此藥物最常見的 品牌名為力得（Lipo-Dox®，由台灣東洋藥品工業股份有限公司 獨家製造及銷售），及一類似產品，已知為在美國的都可喜 (Doxil®)。脂質體多索如必辛（doxorubicin)常常用於治療特 定的癌症，例如乳癌、卵巢癌，以及一種稱為與AIDS相關之 Kaposi’s惡性腫瘤的惡性腫瘤。 對製藥工業而言，製備脂質體藥物是困難的。許多用於製備 脂質體及/或脂質體藥物的方法已被揭露，例如Szoka教授的發 明。 授與 Francis C· Szoka，Jr·之美國專利第 5,077,057、5,277,914 及5,549,910號揭露一種用於製備脂質體懸浮液的方法，該脂質 1270379 體具有所界定之驗尺寸且包封有—有㈣化合物，該化合物於 水:醇類«化財解度不佳。此轉度不_化合物以及 足量的適當脂_容於例如D_之非質子性溶劑中，若有需要 的話，可包含-簡量的舰_ (例如乙醇），錢接著將混 合物經擠壓姐魅―賴㈣核财。所得_質體懸浮液 可、、、二透析或牆，若有需要的話。顯可彻注㈣、多孔盤或 贿其他具有約_醜㈣5麵孔洞尺寸的適當裝置來進 行在此一件美國專利的實施例2中，多素如必辛（d〇x〇r薇化） 係溶解於DMSO中，並添加含有印磷脂酿甘油（EpG) ··㈣脂 :膽驗（EPC) ·•翻醇（Chd) (7:3:6)的乙醇溶液内。脂 質體係藉由將脂質-多索如必辛（dG_bidn)混合物，在贼 下，注射至一由 140mMNaCl_10mMTriS_HCl (ΡΗ4·0)組成的水 相中。將脂質體紐析，並藉由;I；柱層析法，將經脂質體包 封的多索如必辛（doxorubicin)與未被包封的物質分離。所得的 脂質體的顆粒直徑為277 nm，以及41.2%的多索如必辛 (doxorubicin)係包封在脂質體顆粒内。 然而，前述方法無法弭除某些問題，例如毒性有機溶劑、無 菌條件的維持，以及脂質體的均一性及品質，因此前述方法不適 合用於大規模製造。 習知技術已揭露一種利用脂質體多索如必辛（doxorubicin) 治療胰臟癌的方法，例如“評估康利斯（CAELYX，脂質體多 索如必辛（d〇x〇rubicin)，都可喜（Doxii))在治療無法切除之 胰臟癌之耐受性及功效的第II期研究” （Ann Oncol. 2001 〇ct ; 12(10) :1399_402)。然而，此研究得到CAELYX® (脂質 1270379 體多索如必辛（dox〇rubicin)，都可喜（Doxil®))未見客觀反 應的結論。然而，所使用之療程無法有效地治療此疾病。 本發明除了提供適於工業製造之製備脂質體多索如必辛 (doxorubicin)的方法之外，亦提供利用脂質體多索如必辛 (doxorubicin)治療胰臟癌的方法。為了評估功效，研究經過及 未經過脂質體多索如必辛（doxorubicin)投藥之帶有胰臟腫瘤的 大白鼠的腫瘤尺寸及存活時間。 所有上述文獻及專利，以及下文中所提及的文獻，皆全文併 入本說明書甲以供參考。 [發明内容] 因此，本發明之一目的為製造一脂質體多索如必辛 (doxorubicin)藥學組成物以及有效地使用此組成物來治療胰臟 癌。 在一方面，本發明提供一種治療患有胰臟癌之哺乳動物的方 法，包含投藥一治療上有效量之脂質體多索如必辛（doxorubicin) 藥學組成物的步驟。 在另一方面，本發明提供一種製造脂質體多索如必辛 (doxorubicin)藥學組成物的方法。 [實施方式] 本發明之較佳實施例的詳細說明 本發明提供一種製造脂質體多索如必辛（doxorubicin)的方 法，包含下述步驟··（a)在約55-65°C下，提供包含下述化合 物之預混合物：40-70%二硬脂醯基磷脂醯膽鹼（DSPC)、10-30 %膽固醇，以及15-30%甲氧基-聚乙二醇-二硬脂醯基磷脂醯乙 1270379 醇胺（mPEG-DSPE)與一醇類溶劑之預混合物/醇類溶液，其 中該預混合物與醇類溶劑的比例為約1 : 5 (w/v) ; (b)將預 混合物/醇類溶液與0.2-0.8 N硫酸銨溶液，以約1: 2-10( v/v) 的比例混合，以形成一混合物；（c)在50_7(rCT，較佳為在60 C下’使步驟（b)所得的混合物經過〇·〇5-〇·45μιη之孔洞的孔 洞擠壓處理，以形成一預脂質體懸浮液；（d)在室溫下，利用5 %至15%蔗糖水溶液，透析該預脂質體懸浮液，以致能獲得脂 質體懸浮液；以及（e)在55-65°C下，較佳為在60°C下，將多 索如必辛（doxorubicin)與步驟（d)自蔗糖水溶液所獲得的該 脂質體懸浮液混合。 當與習知方法比較時，本發明可在低壓下（約40至140 psi) 下進行，但得到較高的產率（約2至10 L/min)，其比在約1〇〇 至200 psi下進行且產率為1至5 L/min的習知方法好。 利用力得（Lipo_Dox®)，以靜脈注射（I.V·)的方式治療 患有胰臟癌的大白鼠，劑量為10 mg/kg。結果顯示於下述說明 中〇 經過生理食鹽水或力得（Lipo-Dox®)投藥後，體重、腫瘤 尺寸及存活時間的改變係顯示於表1及2中。 第1圖顯示在投藥劑量為10 mg/kg生理食鹽水（對照組） 或力得（Lipo_Dox®)後，相對腫瘤體積隨著時間的改變。治療 是在將腫瘤植入大白鼠後第〇天及第13天開始。大白鼠是在第 0、3、7及10天時，利用生理食鹽水或力得（Lipo-Dox®) (1〇 mg/kg)治療。依所指定的天數測量腫瘤。以Γ/%指數表示相 對腫瘤體積，其中K為所指定天數的腫瘤體積，以及K為治療 1270379 開始時同一腫瘤的體積。 第2圖顯示在投藥劑量為10mg/kg生理食鹽水（斜昭故）， (Lipo-Dox®) ^0 入大白鼠後第0天及第13天開始。大白鼠是在第〇、3、7及⑴天护 利用生理食鹽水或力得（LiP〇-D〇X®) (10mg/kg)治療。丁 表1於經過或未經過力得（Lipo-Dox®)治療後，體重的改變 _ _體重(g)_ 編號 注射前 注射後 --- -----— 3天 7天 10天 14天 17 天 21 夭：Μ 1 生理食鹽水組 303 180 180 220 230 死亡 死亡 死亡 死亡 308 180 180 240 225 240 死亡 死亡 死亡 力得（LiPODOX)組 304 170 180 170 120 110 110 110 110 305 180 190 180 120 110 110 110 110 311 160 170 170 120 110 110 110 110 表2 於經過或未經過力得（Lipo-Dox®)治療後，腫瘤體積及 存活時間的改變 1270379 腫瘤體積(mm3) 編號 注射前 注射後 減少或增 加％ 反應 存活時 間(d) 3天 7天 10天 生理食鹽水組 303 397.55 2771.72 6885.52 11258.26 增加 391.92% 不良 13 308 595.73 3934.75 7601.64 12050.18 增加 291.11% 不良 15 力得（Lipo-Dox®) 組 304 774.04 1195.91 318.21 0 減少 100% 良好 26 305 785.7 671.62 338.2 0 減少 100% 良好 27 311 727.59 1139.37 397.89 0 減少 100% 良好 27 當在第0、3、7及10天靜脈注射時，力得（Lipo-Dox®) 對抗皮下（sx·)植入的AR43J胰臟腫瘤是高度有效的。在10 mg/kg之劑量下，所有動物皆治癒，以及經治療的大白鼠的存活 率呈現增加（參見第1圖及第2圖）。於經由靜脈注射四次投藥 生理食鹽水後，大白鼠的體重穩定地增加，然而，利用力得 (Lipo-Dox®)治療的大白鼠之體重呈現下降高達33.33百分 比。此結果可得到下述結論：力得（Lipo-Dox®)的投藥為胰臟 癌的有效治療方法。 實施例 下述實施例例示說明製備、特性描述及使用本發明之組成物 的方法。此等實施例並非意欲限制本發明之範圍。 實施例1脂質體多索如必辛（多索如必辛（doiombicirO)華學 組成物，力得（Lipo-Dox®)的製備 1270379 將 16.8 克之 peg_2〇〇〇-DSPE (購自 Genzyine Co·，美國）、 27·4克之膽固醇（購自N〇F c〇·，日本）以及38·2克之dSPC (購自NOF Co·，曰本）添加至玻璃容器内之600毫升乙醇中。 在60 C下使混合物混合良好。當持續地攪拌混合物並將混合物 維時在60°C，將4升之硫酸敍水溶液添加至混合物中。在此溫 度下，乙醇幾乎全部蒸發。對混合物進行孔洞擠壓處理，使用 1·5升之渡膜（購自T〇y〇Kaisha，Ltd·，日本）。孔洞擠壓處理包 含： (1 )使用第一過濾膜（142毫米，0.1微米），將混合物過 濾10次；以及 (2 )使用第二過濾膜（142毫米，〇·〇5微米），將混合物 再過遽10次。 擠壓壓力係保持在3至1〇公斤/平方公分，以及流速為約 2至10升/分鐘。收集4500毫升濾液並接著利用3〇升(w /w)之蔗糖溶液透析，該蔗糖溶液係製備於3〇 中空纖維中 (A/G Technology，UFP-30心6Α，3〇，_ 麵，似⑻平方公 分）。殘留的乙醇係藉由透析去除。所收集的溶液的體積約3〇〇〇 笔升，以及所收集的溶液為不含有乙醇的脂質體懸浮液。 將所製造之3000笔升的脂質體懸浮液添加至含有8_毫克 多索如必辛（cb_bidn)鹽酸鹽（紅色粉末）的玻璃容器中， 接著持續地添加預先製備的細毫升組胺酸—餘溶液。將混 合物置於6(TC的水浴中並獅3G a鐘，接著冷卻至約机， 利用9%之蔗糖溶液稀釋至4升，並混合良好。 將產物包裝在用於作為可注射製劑的無菌玻璃小管瓶中，該 1270379 製劑包含2.0¾克多索如必辛（d〇x〇rubicin)鹽酸鹽/毫升。 實施例2 製備ΑΙΜΖΓ細啤懸浮液 衍生型且為重氮絲胺酸引發之大白鼠胰臟腫瘤的AR42J細 胞株（ATCC，CRL 1492)係購自ATCC。在37°C下以及95% 氧氣及5%二氧化碳的濕度環境下，將細胞係培養在 Ham’sF_12K (GibcoBRL)培養基中，該培養基含有20%胎牛 白蛋白、2mM之L-谷胺醯胺，以及ι·5克/升之碳酸氫鈉。細 胞例行地以lx 1〇6濃度培養在Τ-燒瓶中。藉由主要培養，利用 胰蛋白酶-EDTA (GibcoBRL)收取藉由細胞，以及在lOOOrpm 下離心細胞懸浮液1〇分鐘，並調整至1χ 1〇7細胞/毫升。使用 錐蟲藍（Trypanblue)來評估AR42J胰臟腫瘤細胞的生命力。 實施例3 植入AR42J胰臟癌細胞 使用5隻路易士大白鼠(Lewisrats，由台灣國家科學會提供） 來進行實驗，每隻大白鼠重約160克至18〇克，以及所有大白 鼠皆約3週大。食物及水係無限制地供給。動物照料及利用程 序係根據原子能研究委員會的實驗室動物的照料及利用指引 (Guide for the Care and Use of Laboratory Animals from the1270379 发明, DESCRIPTION OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to a method for producing a pharmaceutical composition of liposomal doxorubicin, and a use thereof for treating pancreatic cancer. [Prior Art] In 2003, in the United States, approximately 30,000 patients died of pancreatic cancer, and approximately 30,700 new cases were diagnosed, making pancreatic cancer one of the ten most common causes of cancer-related mortality. For many patients, surgery, radiation therapy, and chemotherapy are treatment options for life-threatening and/or symptom-relieving, but rarely cured (Cancer Facts and Figures, American Cancer Society 2003). Compared with non-liposomal drugs, liposomal drugs can reach the primary tumor and its metastasis quite successfully after intravenous injection into animals and humans, and show a strong therapeutic effect, as well as mild side effects. One of the most widely developed liposome drugs is the liposome doxorubicin. In Taiwan, the most common brand name for this drug is Lipo-Dox®, which is manufactured and sold exclusively by Taiwan Toyo Pharmaceutical Industries Co., Ltd., and a similar product known as Doxil® in the United States. . The liposomal doxorubicin is often used to treat specific cancers, such as breast cancer, ovarian cancer, and a malignant tumor called Kaposi's malignant tumor associated with AIDS. For the pharmaceutical industry, the preparation of liposome drugs is difficult. A number of methods for preparing liposomal and/or liposome drugs have been disclosed, for example, by Professor Szoka. A method for preparing a liposome suspension having a defined size and encapsulating a compound having (iv) is disclosed in U.S. Patent Nos. 5, 077, 057, 5, 277, 914, and 5, 549, 910 to the disclosure of U.S. Pat. The compound is in water: alcohol «decomposition is not good. This rotation is not a compound and a sufficient amount of the appropriate fat is contained in an aprotic solvent such as D_, if necessary, may contain - a simple ship _ (such as ethanol), the money is then squeezed Sister charm - Lai (four) nuclear wealth. The resulting plastid suspension can be, dialysis or wall, if necessary. It is obvious that (4), a porous disk or a bribe other suitable device having a size of about _ ugly (four) 5 face holes is carried out in the embodiment 2 of this U.S. patent, which is a multi-sugar (d〇x〇r Weihua) It is dissolved in DMSO and added to an ethanol solution containing phosphatidylcholine (EpG) · (iv) lipid: biliary test (EPC) · decyl alcohol (Chd) (7:3:6). The lipid system was injected into an aqueous phase consisting of 140 mM NaCl 10 mM TriS_HCl (ΡΗ4·0) by a mixture of lipid-dose bis (dG_bidn) under thief. The liposomes were subjected to neoprecipitation and the liposomally encapsulated doxorubicin was separated from the unencapsulated material by column chromatography. The resulting liposomes had a particle diameter of 277 nm, and 41.2% of doxorubicin was encapsulated within the liposome particles. However, the foregoing methods are not capable of eliminating certain problems such as toxic organic solvents, maintenance of sterile conditions, and homogeneity and quality of liposomes, and thus the foregoing methods are not suitable for large-scale production. Conventional techniques have disclosed a method for treating pancreatic cancer using the liposome doxorubicin, for example, "evaluation of CAELYX (liposome dorsine (d〇x〇rubicin) is pleasing ( Doxii)) Phase II study in the treatment of unresectable pancreatic cancer tolerance and efficacy" (Ann Oncol. 2001 〇 ct; 12(10): 1399_402). However, this study yielded the conclusion that CAELYX® (Dolly 1270379, dox〇rubicin, Doxil®) did not show an objective response. However, the course of treatment used does not effectively treat the disease. In addition to providing a method for preparing liposomal doxorubicin suitable for industrial production, the present invention also provides a method for treating pancreatic cancer using the liposome doxorubicin. To assess efficacy, tumor size and survival time of pancreatic tumor-bearing mice that had been administered without and without liposomal doxorubicin were studied. All of the above documents and patents, as well as the documents mentioned hereinafter, are hereby incorporated by reference in their entirety. SUMMARY OF THE INVENTION Accordingly, it is an object of the present invention to produce a liposomal doxorubicin pharmaceutical composition and to effectively use the composition to treat pancreatic cancer. In one aspect, the invention provides a method of treating a mammal having pancreatic cancer comprising the step of administering a therapeutically effective amount of a liposomal doxorubicin pharmaceutical composition. In another aspect, the invention provides a method of making a liposomal doxorubicin pharmaceutical composition. [Embodiment] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE INVENTION The present invention provides a method for producing a liposome, doxorubicin, comprising the following steps: (a) at about 55-65 ° C, A premix comprising the following compounds: 40-70% distearyl phospholipid choline (DSPC), 10-30% cholesterol, and 15-30% methoxy-polyethylene glycol-distearate a premix/alcohol solution of a phospholipid Ethylamine 1270379 alkanolamine (mPEG-DSPE) with an alcohol solvent, wherein the ratio of the premix to the alcohol solvent is about 1:5 (w/v); (b) The premix/alcohol solution is mixed with a 0.2-0.8 N ammonium sulfate solution at a ratio of about 1:2-10 (v/v) to form a mixture; (c) at 50-7 (rCT, preferably at 60 C) Next, the mixture obtained in the step (b) is subjected to pore extrusion through a hole of 〇·〇5-〇·45 μιη to form a pre-liposome suspension; (d) at room temperature, using 5% to 15% An aqueous solution of sucrose, dialysis of the pre-liposome suspension to obtain a liposome suspension; and (e) at 50-65 ° C, preferably at 60 ° C, doxorubib (doxorub) Icin) is mixed with the liposome suspension obtained from the aqueous sucrose solution in step (d). When compared to conventional methods, the invention can be carried out at low pressure (about 40 to 140 psi), but with higher yields. Rate (about 2 to 10 L/min), which is better than the conventional method of performing at about 1 to 200 psi and yielding 1 to 5 L/min. Using Lipo_Dox® for intravenous injection The method of (IV·) was used to treat rats with pancreatic cancer at a dose of 10 mg/kg. The results are shown in the following instructions: body weight, tumor after administration of physiological saline or Lipo-Dox® Changes in size and survival time are shown in Tables 1 and 2. Figure 1 shows the relative tumor volume over time after administration of 10 mg/kg saline (control) or Lipo_Dox®. The treatment was started on the third day and the 13th day after the tumor was implanted into the rats. The rats were treated with physiological saline or Lipo-Dox® on days 0, 3, 7 and 10. 1〇mg/kg) treatment. Tumors were measured according to the number of days specified. Relative tumor volume was expressed as Γ/% index, where K is the specified number of days The tumor volume, and K is the volume of the same tumor at the start of treatment 1270379. Figure 2 shows that the dose of 10 mg/kg of physiological saline (inclined), (Lipo-Dox®) ^0 after entering the mouse 0 Days and Day 13. The rats were treated with physiological saline or LiP〇-D〇X® (10 mg/kg) on Days 3, 7, and (1). Ding Table 1 changes in body weight after treatment with or without Lipo-Dox® _ _ weight (g) _ number before injection before injection ----- ----- 3 days 7 days 10 days 14 Day 17 Day 21 夭: Μ 1 Physiological saline group 303 180 180 220 230 Death death death 308 180 180 240 225 240 Death death death force (LiPODOX) group 304 170 180 170 120 110 110 110 110 305 180 190 180 120 110 110 110 110 311 160 170 170 120 110 110 110 110 Table 2 Changes in tumor volume and survival after treatment with or without Lipo-Dox® 1270379 Tumor volume (mm3) Number decreased after injection before injection Or increase % reaction survival time (d) 3 days 7 days 10 days physiological saline group 303 397.55 2771.72 6885.52 11258.26 increase 391.92% bad 13 308 595.73 3934.75 7601.64 12050.18 increase 291.11% bad 15 Lipo-Dox® group 304 774.04 1195.91 318.21 0 reduction 100% good 26 305 785.7 671.62 338.2 0 reduction 100% good 27 311 727.59 1139.37 397.89 0 reduction 100% good 27 when on the 0th, 3rd, 7th and 10th day Injection, to give a force (Lipo-Dox®) against subcutaneously (sx ·) implanted AR43J pancreatic tumors are highly effective. At 10 mg/kg, all animals were cured and the survival of treated rats increased (see Figures 1 and 2). The body weight of the rats was steadily increased after intravenous administration of the physiological saline four times. However, the body weight of the rats treated with Lipo-Dox® decreased by as much as 33.33. This result leads to the conclusion that administration of Lipo-Dox® is an effective treatment for pancreatic cancer. EXAMPLES The following examples illustrate the preparation, characterization, and methods of using the compositions of the present invention. These examples are not intended to limit the scope of the invention. Example 1 Preparation of liposomal polysozoic ruthenium (doiombicirO), preparation of Lipo-Dox® 1270379 16.8 g of peg_2〇〇〇-DSPE (purchased from Genzyine Co· , USA), 27. 4 grams of cholesterol (purchased from N〇F c〇·, Japan) and 38.2 grams of dSPC (purchased from NOF Co., 曰本) were added to 600 ml of ethanol in a glass container. The mixture was well mixed under C. When the mixture was continuously stirred and the mixture was maintained at 60 ° C, 4 liters of a sulfuric acid aqueous solution was added to the mixture. At this temperature, almost all of the ethanol was evaporated. For the treatment, a 1.5 liter film (available from T〇y〇 Kaisha, Ltd., Japan) was used. The hole extrusion treatment consisted of: (1) using a first filtration membrane (142 mm, 0.1 μm) to filter the mixture 10 times; and (2) using a second filter membrane (142 mm, 〇·〇 5 μm), the mixture was further tween 10 times. The extrusion pressure was maintained at 3 to 1 kg/cm 2 and the flow rate was about 2 to 10 liters / minute. Collect 4,500 ml of filtrate and then use 3 liters (w /w) dialysis of sucrose solution prepared in 3 〇 hollow fiber (A/G Technology, UFP-30 core 6 Α, 3 〇, _ surface, like (8) cm ^ 2 ). Residual ethanol by dialysis The volume of the collected solution was about 3 liters, and the collected solution was a liposome suspension containing no ethanol. The 3000 liters of liposome suspension produced was added to contain more than 8 mg. In a glass container of sobbisbine (cb_bidn) hydrochloride (red powder), the previously prepared fine milliliter of histidine-residue solution was continuously added. The mixture was placed in a 6 (TC water bath and lion 3 G a clock) , then cooled to about 10,000 liters, diluted to 4 liters with 9% sucrose solution, and mixed well. The product was packaged in a sterile glass vial for use as an injectable preparation containing 2.03⁄4 gram of Doxor Indole (d〇x〇rubicin) hydrochloride/ml. Example 2 Preparation of AR42J cell line (ATCC, CRL 1492) purchased from the squid suspension-derived type and which is a diazo seric acid-induced pancreatic tumor of rats. From ATCC. At 37 ° C and 95% oxygen and 5 In a humidity environment of % carbon dioxide, the cell line was cultured in Ham's F_12K (GibcoBRL) medium containing 20% fetal bovine albumin, 2 mM L-glutamine, and 1 MPa/liter of hydrogen carbonate. Sodium was routinely cultured in a Τ-flask at a concentration of lx 1 〇 6. By primary culture, trypsin-EDTA (GibcoBRL) was used to collect the cells, and the cell suspension was centrifuged at 1000 rpm for 1 minute, and Adjust to 1χ1〇7 cells/ml. Trypanblue was used to assess the viability of AR42J pancreatic tumor cells. Example 3 Implantation of AR42J pancreatic cancer cells Five Lewis rats (Lewisrats, provided by the National Science Society of Taiwan) were used for experiments. Each rat weighed about 160 to 18 grams, and all rats were approximately 3 weeks old. Food and water are supplied without restriction. Animal Care and Utilization Programs are based on the guidelines for the care and use of laboratory animals by the Atomic Energy Research Council (Guide for the Care and Use of Laboratory Animals from the
Institute of Nuclear Energy Research)。一百萬個 AR42J 細胞係懸 浮在0·1耄升培養基中，並皮下注射至大白鼠的右下肢。植入二 週後，藉由公式：長度（毫米）X寬度2 (毫米2 ) / 2，估算 腫瘤體積。Vt/VG指數表示相對腫瘤體積，其中vt為在所指定 的天數測置的腫瘤體積，以及V〇為在測量開始時同一腫瘤的體 積。 f施例4 投藥力得（Lipo-Dox®)的治唪钕爭 12 Ϊ270379 將帶有AR42J胰臟腫瘤的大白鼠分為二組，其中二隻大白 氣為對照組’以及另外三隻為力得（你〇彻的組。此二組的 大白氣係分別錄生理食鹽水或力得，投藥量為5 至^毫克/公斤，較佳為1〇 $克/公斤），每週二次^星期 -及星期六），共靜脈注射四次。在投藥後需立即小心地觀察， 例如腫瘤體積、體重、活動力程度，以及毛髮掉落情況，並監 測直至沒有大白鼠存活。 现 雖然本發明已連同所謂最佳實施具體例來說明，但應瞭解 =，本發明並不限於上述的具體例，且纽涵蓋包括於本發敗 # 最廣解釋的精神及範圍中的任何不同的改良。 [圖式簡單說明] 第1圖顯示力得（Lipo_Dox@)對帶有趣2;胰臟腫瘤之大 白鼠在腫瘤體積方面的效果；以及 顯示力得（Lipo-Dox®)對帶有AR42J騰臟腫瘤之大 白鼠在存活時間方面的效果。 [圖式之主要元件代表符號表] 鲁 13Institute of Nuclear Energy Research). One million AR42J cell lines were suspended in 0.1 ml of medium and injected subcutaneously into the right lower extremity of rats. Two weeks after implantation, the tumor volume was estimated by the formula: length (mm) X width 2 (mm 2 ) / 2. The Vt/VG index represents the relative tumor volume, where vt is the tumor volume measured over the specified number of days, and V〇 is the volume of the same tumor at the beginning of the measurement. f Example 4 Administration of Lipo-Dox® 12 Ϊ 270379 Rats with AR42J pancreatic tumors were divided into two groups, two of which were the control group and the other three were (You have a thorough group. The two groups of large white gas are recorded with physiological saline or force, the dosage is 5 to ^ mg / kg, preferably 1 〇 $ g / kg), twice a week ^ Week-and-Saturday, a total of four intravenous injections. Immediate attention should be paid immediately after administration, such as tumor volume, body weight, degree of activity, and hair loss, and monitored until no rats survive. Although the present invention has been described in connection with the so-described preferred embodiments, it should be understood that the invention is not limited to the specific examples described above, and that the present invention encompasses any differences in the spirit and scope of the present invention. Improvement. [Simple diagram of the diagram] Figure 1 shows the effect of Lipo_Dox@ on the tumor volume with interesting 2; pancreatic tumors; and Lipo-Dox® pair with AR42J The effect of tumor white rats on survival time. [The main components of the diagram represent the symbol table] Lu 13