TW200835507A

TW200835507A - Compositions including triciribine and epidermal growth factor receptor inhibitor compounds or salts thereof and methods of use thereof

Info

Publication number: TW200835507A
Application number: TW096146309A
Authority: TW
Inventors: Lawrence Akinsanmi
Original assignee: Vioquest Pharmaceuticals Inc
Priority date: 2006-12-05
Filing date: 2007-12-05
Publication date: 2008-09-01
Also published as: WO2008070100A8; WO2008070100A1

Abstract

This application relates to combination therapies including triciribine compounds and epidermal growth factor receptor inhibitor compounds, particularly erlotinib-like compounds and compositions with reduced toxicity for the treatment and prevention of tumors, cancer, and other disorders associated with abnormal cell proliferation.

Description

200835507 九、發明說明·· d日月所屬之技術領域】本案請求美國臨時專利申請案第60/872,778號，申請曰 2006年12月5日之權益，該案全文以引用方式併入此處。 5 發明領域本案係有關具有較低毒性，用於治療及預防腫瘤、癌症及其它與異常細胞增生相關之病症之組合治療，該組合 /口療包括曲西立濱（triciribine)化合物及表皮生長因子受體抑制劑化合物特別為爾洛堤尼(erl〇tinib)系列化合物及組成 10 物。 L· lltr ^ 發明背景癌症為細胞之異常生長。儘管受到空間限制、與其它細胞共享營養素，或由身體發送來停止繁殖的信號，癌細 15 胞快速複製。癌細胞經常與正常細胞有差異分享，無法適當發揮功能，且可能傳播至身體的多個區域。組織的異常生長稱作為腫瘤為可未經控制的生長及***之細胞團簇。腫瘤可能為良性（非癌性)腫瘤或惡性(癌性)腫瘤。良性腫瘤的生長緩慢且不會傳播。惡性腫瘤可快速生長、入侵且摧 20 毀鄰近的異常組織且傳播遍及身體。痴症可根據其來源體液種類或組織種類而歸類，或可根據癌症於身體中首次出現的位置歸類。此外，若干癌症為混合型。癌症可分成五大類，亦即癌瘤、肉瘤、淋巴瘤、白血病、及骨髓瘤，指示癌症之組織歸類及血液歸類。癌 5 200835507 瘤為身體組織中稱作為上皮組織，上皮組織覆蓋或襯墊於器官、腺體或身體結構表面之癌症。舉例言之，胃内襯的癌症稱作為癌瘤。多種癌瘤影響涉及分泌之器官或腺體，諸如製造乳汁的乳腺。癌瘤占全部癌症病例之80%至90%。 5 肉瘤為由結締組織諸如軟骨、脂肪、肌肉、肌腱、及硬骨所長出的惡性腫瘤。最常見的肉瘤是長在硬骨上的腫瘤，常見於年輕成人。肉瘤之實例包括骨肉瘤（硬骨)及軟骨肉瘤 (軟骨）。淋巴瘤係指源自於淋巴系統的淋巴結或淋巴腺之癌症，淋巴結或淋巴腺係負責製造白血球且清潔體液，或淋 10 巴瘤係源自於器官諸如腦部及乳腺。淋巴瘤被分成兩大類：何杰金氏淋巴瘤及非何杰金氏淋巴瘤。白血病也稱作為血癌是一種骨髓的癌症，造成骨髓無法製造正常的紅血球及白血球及血小板。白血球為對抗感染所需。紅血球為預防貧血所需。血小板可保持身體避免瘀青及出血。白血 15 病之實例包括急性骨體性白血病、慢性骨體性白血病、急性淋巴細胞性白血病、及慢性淋巴細胞性白血病。骨聽性及淋巴性等詞係指涉及的細胞來源。最後，骨髓瘤係於骨髓之漿細胞生長。於某些病例中，骨髓瘤細胞集合於一根硬骨而形成單一腫瘤稱作為漿細胞瘤。但於其它病例中， 20 骨髓瘤細胞集合於多根硬骨，形成多個骨腫瘤。稱作為多發性骨髓瘤。腫瘤誘導及進行經常係由於腫瘤細胞基因體中累積的變化的結果。此種變化可能包括細胞生長抑制因子或腫瘤阻遏子基因的去活化，以及包括細胞生長促進基因或致癌 200835507 基因的活化。今日於動物模型中已經識別出數百種活化的細胞致癌基因，但已經證實只有極少數基因係與人類癌症相關（Weinberg等人，致癌基因及癌症之分子緣起，1989，冷泉港實驗室，紐約；Stanbridge J·等人，細胞，1990, 63: p 5 867-874; Godwin等人，婦科腫瘤學：原理與實務，Hoskins W.J·，Perez C.A·及Young R.C·(編輯），1992· ρρ 87-116，里平可特(Lippincott)，費城）。於人類癌症中致癌基因的活化可能來自於諸如基因複本數目的增加或結構改變等因素。此等因素造成多項細胞效應，例如此等因素可能導致基因 10產物的過度表現。若干涉及人類癌症之致癌基因可經由基因過度表現而活化。顯然由癌症基因所獲得之連續基因失序，導致控管正常細胞增生、分化及經過規劃的細胞凋亡之調節信號轉導回路的缺陷（Hanahan，D·及Weinberg R.A·，細胞，2000. 15 1〇0(丨）：P· 57_700)。如此又導致細胞生理的基本缺陷造成惡性病。此等缺陷包括：a)生長信號之自我不足（亦即生長因子受體酪胺酸激酶諸如EGFR的過度表現，及下游信號轉導徑路諸如 Ras/Raf/Mek/Erk 1/2 及 Ras/PI3K/Akt 之失序活化），b)對抗生長信號有抗性（亦即TGF/5及其受體之表現降 20低），c)迴避細胞凋亡（亦即前細胞凋亡p53之喪失；前存活 Bcl-2之過度表現；存活徑路諸如PI3K/Akt所媒介之存活徑路的過度活化），d)持續血管新生（亦即veGF之分泌濃度高）以及f)組織入侵及轉移（亦即胞外蛋白酶及前轉移接合素) (Hanahan，D·及 Weinberg R.A.，細胞，2000· 100(1): ρ· 7 200835507 57-700)。受體酪胺酸激酶諸如EGFR、ErbB2、VEGFR及胰島素系列生長因子I受體（IGF-1R)密切涉及多種人類癌症的發展’包括大腸直腸癌、胰癌、乳癌、及卵巢癌（Khaleghpour 5 K專人’癌症發生 ’ 2004.25(2); ρ·241·8 ; Sekharam Μ·等人，癌症研究，2003, 63(22): ρ·7708-16)。配體諸如EGF、VEGF 及IGF-1結合至其受體’促成特有赂胺酸激酶活性的刺激，於受體之胞質功能部位的特定酪胺酸的自行磷酸化以及觸發多種不同複雜的信號轉導徑路之發訊蛋白質之募集 10 (Olayioye M.A.等人，Embo J，2000. 19(13): ρ· 3159-67 ; Porter A.C·及Vaillancourt R.R.，致癌基因，1998.17(11 綜論）: ρ· 1343-52)。如此又導致多種腫瘤存活徑路及腫瘤發生徑路諸如Ras/Raf/Mek/Erk 1/2、JAK/STAT3及PI3K/Akt徑路之活化。雖然全部三個徑路皆暗示與大腸癌、胰癌、乳癌及卵 15 巢癌之腫瘤發生有關，但藉Akt所媒介之徑路業已顯示於惡性轉形之多個步驟有關鍵重要性，惡性轉形之步驟包括細胞增生、抗細胞凋亡/存活、入侵及轉移及腫瘤新生（Datta S.R.等人，基因 Dev，1999.13(22): ρ·2905-27)。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。。 5 FIELD OF THE INVENTION The present invention relates to combination therapy with lower toxicity for the treatment and prevention of tumors, cancer and other disorders associated with abnormal cell proliferation, including triciribine compounds and epidermal growth factors. The receptor inhibitor compound is specifically a compound of the erlotinib series and a composition of 10. L·lltr ^ Background of the invention Cancer is the abnormal growth of cells. Cancer cells replicate rapidly despite space constraints, sharing nutrients with other cells, or signals sent by the body to stop reproduction. Cancer cells often share differences with normal cells, fail to function properly, and may spread to multiple areas of the body. Abnormal growth of tissue is referred to as a cluster of cells in which the tumor is uncontrolled for growth and division. The tumor may be a benign (non-cancerous) tumor or a malignant (cancerous) tumor. Benign tumors grow slowly and do not spread. Malignant tumors can rapidly grow, invade and destroy 20 adjacent abnormal tissues and spread throughout the body. A disease can be classified according to the type of body fluid or tissue it is derived from, or it can be classified according to the location where the cancer first appears in the body. In addition, several cancers are mixed. Cancer can be divided into five categories, namely cancer, sarcoma, lymphoma, leukemia, and myeloma, indicating tissue classification and blood classification of cancer. Cancer 5 200835507 Tumor is a cancer in the body tissue called epithelial tissue that covers or pads the surface of organs, glands or body structures. For example, a stomach-lined cancer is referred to as a cancer. A variety of cancerous effects involve organs or glands that are secreted, such as the mammary glands that make milk. Cancers account for 80% to 90% of all cancer cases. 5 Sarcomas are malignant tumors that are formed by connective tissues such as cartilage, fat, muscles, tendons, and hard bones. The most common sarcoma is a tumor that grows on a hard bone and is common in young adults. Examples of sarcomas include osteosarcoma (hard bone) and chondrosarcoma (cartilage). Lymphoma refers to cancer of the lymph nodes or lymph nodes derived from the lymphatic system. The lymph nodes or lymph nodes are responsible for making white blood cells and cleaning body fluids, or the tumors are derived from organs such as the brain and breast. Lymphomas are divided into two broad categories: Hodgkin's lymphoma and non-Hodgkin's lymphoma. Leukemia, also known as blood cancer, is a bone marrow cancer that causes the bone marrow to fail to produce normal red blood cells and white blood cells and platelets. White blood cells are needed to fight infection. Red blood cells are needed to prevent anemia. Platelets keep the body from dark and bleeding. Examples of white blood 15 diseases include acute osteomyelitis, chronic osteomyelitis, acute lymphocytic leukemia, and chronic lymphocytic leukemia. Words such as osteosynthesis and lymphatics refer to the source of cells involved. Finally, myeloma is caused by the growth of plasma cells in the bone marrow. In some cases, myeloma cells are aggregated into a single bone to form a single tumor called a plasmacytoma. In other cases, however, 20 myeloma cells assemble multiple hard bones to form multiple bone tumors. Called as multiple myeloma. Tumor induction and progression are often the result of changes in the accumulation of tumor cell genomes. Such changes may include deactivation of cytostatic or tumor repressor genes, as well as activation of the cell growth promoting gene or the oncogenic 200835507 gene. Hundreds of activated cellular oncogenes have been identified in animal models today, but only a few genes have been shown to be associated with human cancer (Weinberg et al., the genes of oncogenes and cancer, 1989, Cold Spring Harbor Laboratory, New York) Stanbridge J. et al., Cell, 1990, 63: p 5 867-874; Godwin et al., Gynecologic Oncology: Principles and Practice, Hoskins WJ, Perez CA· and Young RC (eds.), 1992· ρρ 87 -116, Lippincott, Philadelphia). The activation of oncogenes in human cancer may come from factors such as an increase in the number of gene copies or structural changes. These factors contribute to a number of cellular effects, such as these factors may result in excessive expression of the gene 10 product. Several oncogenes involved in human cancer can be activated by excessive gene expression. It is clear that the continuous gene disorder obtained by the cancer gene leads to defects in regulating the regulation of signal transduction pathways in normal cell proliferation, differentiation and planned apoptosis (Hanahan, D. and Weinberg RA., Cell, 2000. 15 1 〇0 (丨): P· 57_700). This in turn leads to a fundamental defect in cell physiology leading to a malignant disease. These defects include: a) self-deficiency of growth signals (ie, excessive expression of growth factor receptor tyrosine kinases such as EGFR, and downstream signal transduction pathways such as Ras/Raf/Mek/Erk 1/2 and Ras/ PI3K/Akt is out of sequence activation), b) resistant to growth signals (ie, TGF/5 and its receptors are reduced by 20), c) avoidance of apoptosis (ie, loss of pre-apoptotic p53; Excessive survival of pre-existing Bcl-2; survival pathways such as excessive activation of the survival pathway mediated by PI3K/Akt), d) sustained angiogenesis (ie high secretion of veGF) and f) tissue invasion and metastasis (also That is, extracellular protease and pre-transfer lignin) (Hanahan, D. and Weinberg RA, Cell, 2000·100(1): ρ·7 200835507 57-700). Receptor tyrosine kinases such as EGFR, ErbB2, VEGFR and the insulin series growth factor I receptor (IGF-1R) are closely involved in the development of a variety of human cancers including colorectal cancer, pancreatic cancer, breast cancer, and ovarian cancer (Khaleghpour 5 K). Specialized 'cancer occurrence' 2004.25 (2); ρ·241·8; Sekharam Μ· et al, Cancer Research, 2003, 63(22): ρ·7708-16). Ligands such as EGF, VEGF, and IGF-1 bind to their receptors to facilitate stimulation of specific glucuronide kinase activity, self-phosphorylation of specific tyrosine at the cytoplasmic functional site of the receptor and trigger a variety of different complex signals The collection of signaling proteins for transduction pathways 10 (Olayioye MA et al, Embo J, 2000. 19(13): ρ·3159-67; Porter AC· and Vaillancourt RR, oncogenes, 1998.17 (11 summary): ρ· 1343-52). This in turn leads to a variety of tumor survival pathways and tumorigenic pathways such as Ras/Raf/Mek/Erk 1/2, JAK/STAT3 and PI3K/Akt pathways. Although all three pathways are implicated in tumorigenesis of colorectal cancer, pancreatic cancer, breast cancer, and egg 15 nest cancer, the path taken by Akt has been shown to be critical in the multiple steps of malignant transformation, malignant. The steps of transformation include cell proliferation, anti-apoptosis/survival, invasion and metastasis, and tumor neoplasia (Datta SR et al., Gene Dev, 1999. 13(22): ρ. 2905-27).

Akt為絲胺酸/蘇胺酸蛋白質激酶(也稱作為PKB)，Akt 20 有三個家族成員Aktl、Akt2及Akt3。以生長因子或存活因子刺激細胞，結果導致募集至脂質激酶磷酸肌糖苷-3-OH-激酶(PI3K)之受體，PI3K將磷酸肌糖醇-4,5-二磷酸(PIP2)磷酸化成為PIP3，PIPJfAkt募集至漿膜，於漿膜，Akt藉於 Thr308及Ser473 (Aktl)、Thr308及Ser474 (Akt2)及Thr308及 200835507Akt is a serine/threonine protein kinase (also known as PKB), and Akt 20 has three family members Aktl, Akt2 and Akt3. Stimulation of cells with growth factors or survival factors results in the recruitment of a receptor for phosphokinase phosphoinositide-3-OH-kinase (PI3K), which phosphorylates phosphoinositide-4,5-diphosphate (PIP2) into PIP3, PIPJfAkt is recruited to the serosa, in the plasma membrane, Akt is borrowed from Thr308 and Ser473 (Aktl), Thr308 and Ser474 (Akt2) and Thr308 and 200835507

Ser472 (Akt3) (Datta S.R·等人，基因Dev，1999.13(22): p.2905-27)之磷酸化活化。如此，PI3K係經由磷酸化PIP2 及PIP2轉化成為PIP3而活化Akt。磷酸酶PTEN將PIP3去磷酸化成為PIP2，如此阻止Akt的活化。 5 大部分人類癌症含有過度活化之Akt(Datta S.R.等人，基因Dev，1999.13(22): ρ·2905-27 ; BellacosaA.等人，國際癌症期刊，1995.64(4): p.280-5 ; Sun M·等人，Am J Pathol， 2001. 159(2)·· ρ· 431-7)。特別分別於57%、32%、27%及36% 人大腸直腸癌、胰癌、乳癌、及卵巢癌中，Akt過度表現及 10 /或高度活化（Roy H.K.等人，癌症發生，2002.23(1): p.201-5 ; Altomare D.A·等人，J細胞Biochem，2003. 88(1): ρ· 470_6 ; Sun Μ·等人，癌症研究，2001.61(16): ρ·5985-91 ; Stal 0·等人，乳癌研究，2003.5(2): ρ· R37-44 ; Cheng J.Q. 等人，Proc Natl Acad Sci USA，1992. 89(19): p. 9267-71; 15 Yuan Z.Q·等人，致癌基因，2000· 19(19): p· 2324-30)。Akt 的過度活化係由於Akt本身之擴增及/或過度表現，以及Akt 的上游之遺傳變化，包括受體酪胺酸激酶及/或其配體的過度表現及磷酸酶PTEN的刪失(Khaleghpour K等人，癌症發生，2004· 25(2); p.241-8 ; Sekharam M.等人，癌症研究， 20 2003，63(22): ρ·7708_16 ; Cohen B.D·等人，Biochem SocPhosphorylation activation of Ser472 (Akt3) (Datta S. R. et al., Gene Dev, 1999. 13(22): p. 2905-27). Thus, PI3K activates Akt by converting PIP2 and PIP2 into PIP3. The phosphatase PTEN dephosphorylates PIP3 to PIP2, thus preventing the activation of Akt. 5 Most human cancers contain over-activated Akt (Datta SR et al., Gene Dev, 1999. 13(22): ρ·2905-27; Bellacosa A. et al., International Journal of Cancer, 1995. 64(4): p.280-5; Sun M. et al., Am J Pathol, 2001. 159(2)·· ρ· 431-7). Particularly in 57%, 32%, 27%, and 36% of human colorectal cancer, pancreatic cancer, breast cancer, and ovarian cancer, Akt was overexpressed and 10/ or highly activated (Roy HK et al., Cancer, 2002.23 (1) ): p.201-5; Altomare DA et al., J Cell Biochem, 2003. 88(1): ρ· 470_6 ; Sun Μ· et al., Cancer Research, 2001.61(16): ρ·5985-91 ; Stal 0· et al., Breast Cancer Research, 2003.5(2): ρ· R37-44 ; Cheng JQ et al., Proc Natl Acad Sci USA, 1992. 89(19): p. 9267-71; 15 Yuan ZQ· et al. Oncogene, 2000· 19(19): p· 2324-30). Overactivation of Akt is due to amplification and/or overexpression of Akt itself, as well as genetic changes upstream of Akt, including overexpression of receptor tyrosine kinase and/or its ligands and censoring of phosphatase PTEN (Khaleghpour) K et al, Cancer, 2004. 25(2); p.241-8; Sekharam M. et al., Cancer Research, 20 2003, 63(22): ρ·7708_16; Cohen BD et al., Biochem Soc

Symp，1998. 63: ρ·199-210 ; Muller W.J.等人，Biochem Soc Symp，1998· 63: p.149-57 ; Miller W.E·等人，】¥如1，1995· 69(7)·· ρ·4390-8 ; SlamonD.J·等人，科學，1987.235(4785): ρ·177_82 ; Andrulis 1丄·等人，J Clin Oncol，1998· 16(4): 9 200835507 P.1340-9)。Akt涉及腫瘤發生的構想證實，已經於臨床前期獲付驗證’顯示Akt之異位表現誘導惡性轉形與促成細胞存活（Sun M·等人，Am J Pathol，2001. 159(2): p· 431-7 ; Cheng J.Q·等人，致癌基因，1997. 14(23): Ρ·2793-801);及Akt徑 5 路的摧毀抑制細胞生長及誘導細胞〉周亡(Jetzt Α·等人，癌症研究，2003.63 (20): ρ·6697-706)。癌症及相關疾病之現行治療之功效有限且有多種嚴重非期望的副作用。儘管已經驗證多種抗癌藥物之臨床功效，但其嚴重系統性毒性經常造成多種有展望之化學治療 10劑的臨床發展中斷。此外，受體酿胺酸激酶諸如EGFR及其配體諸如IGF-1之過度表現、Akt之過度表現及/或pteN的喪失(全部皆導致Akt的過度活化）造成預後不良，對化學治療有抗藥性及癌症病人的存活時間短。目前研究策略係強調於研究有效治療模式且有較低風險。 15 如此，包括曲西立濱（triciribine)化合物及爾洛堤尼 (erlotinib)系列化合物之組合治療有展望可用作為腫瘤、癌症及異常細胞增生之可能治療，同時協同地降低經由目前投予之抗癌化合物所引發之毒性或不良副作用。【^^明内3 20 發明概要本發明提供曲西立濱、磷酸曲西立濱及相關化合物組合一種或多種爾洛堤尼系列化合物之組合治療來於個體治療腫瘤或癌症，同時限制系統性毒性。本發明係基於發現過度表現Akt激酶之腫瘤或癌症對TCn及相關化合物之胞 200835507 毒性效應特別敏感，經由爾洛堤尼系列化合物之組合可獲得協同效果。與先前技術及經驗相反，發明人已經判定如何成功地使用曲西立濱及一種或多種爾洛堤尼系列化合物藉以下一種方式或其組合來治療腫瘤及癌症：⑴將曲西立 5 濱及爾洛堤尼系列化合物投予對曲西立濱化合物或爾洛堤 . 尼系列化合物之敏感度提升之病人；（ii)使用規定之劑量水 ' 平，其可最小化曲西立濱化合物及/或爾洛堤尼系列化合物之毒性，同時仍然具有功效；或(iii)使用所述用藥計劃，其 Γ 可最小化曲西立濱化合物及/或爾洛堤尼系列化合物之毒 10 性。於本發明之一個態樣中，本發明涵蓋一種組成物包括： ⑴式I-IV化合物：Symp, 1998. 63: ρ·199-210; Muller WJ et al., Biochem Soc Symp, 1998· 63: p.149-57; Miller WE· et al., ¥¥1,1995· 69(7)·· ρ·4390-8 ; Slamon D.J. et al., Science, 1987.235 (4785): ρ·177_82 ; Andrulis 1丄· et al, J Clin Oncol, 1998·16(4): 9 200835507 P.1340-9) . The concept of Akt involved in tumorigenesis confirms that it has been verified in the preclinical period to show that ectopic display of Akt induces malignant transformation and contributes to cell survival (Sun M. et al., Am J Pathol, 2001. 159(2): p· 431-7 ; Cheng JQ· et al., Oncogene, 1997. 14(23): Ρ·2793-801); and the destruction of Akt 5 pathway inhibits cell growth and induces cells> Weekly death (Jetzt Α· et al. Cancer Research, 2003.63 (20): ρ·6697-706). Current treatments for cancer and related diseases have limited efficacy and a variety of serious undesired side effects. Although the clinical efficacy of various anticancer drugs has been validated, its severe systemic toxicity often results in a disruption in the clinical development of a number of promising chemotherapeutic agents. In addition, overexpression of receptor tyrosine kinases such as EGFR and its ligands such as IGF-1, overexpression of Akt and/or loss of pteN (all leading to excessive activation of Akt) result in poor prognosis and resistance to chemotherapy. The survival time of medicinal and cancer patients is short. Current research strategies emphasize the study of effective treatment modalities with lower risks. 15 Thus, combination therapy including triciribine compounds and erlotinib series of compounds is expected to be available as a possible treatment for tumors, cancer and abnormal cell proliferation, while synergistically reducing the resistance through current administration. Toxic or adverse side effects caused by cancer compounds. [^^明内3 20 SUMMARY OF THE INVENTION The present invention provides a combination treatment of one or more erlotinib compounds in combination with triclinbine, triclinide and related compounds to treat a tumor or cancer in an individual while limiting systemicity. toxicity. The present invention is based on the discovery that tumors or cancers which overexpress Akt kinase are particularly sensitive to the toxic effects of TCn and related compound cells 200835507, and synergistic effects can be obtained by a combination of the lonotines series of compounds. Contrary to prior art and experience, the inventors have determined how to successfully use trocitabine and one or more of the erlotinib compounds to treat tumors and cancer in one of the following ways: (1) to Quxi Li 5 and The lodottini series of compounds are administered to patients with increased sensitivity to the triclinide or erlotini series; (ii) using the prescribed dose of water, which minimizes the tricinemide compound and / or the toxicity of the erlotini series of compounds, while still having efficacy; or (iii) the use of the drug regimen, which minimizes the toxicity of the triclinide compound and/or the erlotini series of compounds. In one aspect of the invention, the invention encompasses a composition comprising: (1) a compound of formula I-IV:

CHtCHt

11 200835507 CH3 CH|11 200835507 CH3 CH|

其中r2’、r3’及r5’分別為氫、視需要可經取代之磷酸根或膦酸根（包括一磷酸、二磷酸、或三磷酸或安定化磷酸 5 前藥）；醯基（包括低碳醯基）；烷基（包括低碳烷基）；醯胺、績酸S旨包括烧基或芳基烧基；績醯基包括甲石黃醯基及苄基，其中該苯基視需要可經以一個或多個如於此處所述芳基之定義中說明之取代基取代；視需要可經取代之芳基磺醯基；脂質包括磷脂質；胺基酸；碳水化合物；胜肽；或 10 膽固醇；或其它藥學上可接受之離去基，其於活體内可提供一種化合物其中R2’、R3’或R5’分別為Η或一磷酸、二磷酸、或三填酸；其中Rx及Ry分別為氫、視需要可經取代之磷酸根；醯基（包括低碳醯基）；醯胺、烷基（包括低碳烷基）；芳香族、 15 去氧伸烷基諸如聚乙二醇、視需要可經取代之芳基磺醯基；脂質包括磷脂質；胺基酸；碳水化合物；胜肽；或膽固醇；或其它藥學上可接受之離去基；於一個實施例中，該化合物係呈5’-磷醚脂質或5’-醚脂質投藥； 12 200835507Wherein r2', r3' and r5' are respectively hydrogen, optionally substituted phosphate or phosphonate (including monophosphate, diphosphate, or triphosphate or stabilized phosphoric acid 5 prodrug); sulfhydryl (including low carbon) An alkyl group (including a lower alkyl group); a decylamine, a sulphuric acid S is intended to include a decyl group or an aryl group; the fluorenyl group includes a sulfonium group and a benzyl group, wherein the phenyl group can be used as needed One or more substituents as described in the definition of aryl as described herein; optionally substituted arylsulfonyl; lipids including phospholipids; amino acids; carbohydrates; peptides; Cholesterol; or other pharmaceutically acceptable leaving group, which provides a compound in vivo wherein R2', R3' or R5' are respectively hydrazine or monophosphate, diphosphate, or tribasic acid; wherein Rx and Ry are respectively Hydrogen, optionally substituted phosphate; sulfhydryl (including lower sulfhydryl); decylamine, alkyl (including lower alkyl); aromatic, 15 deoxyalkylene such as polyethylene glycol, An arylsulfonyl group which may be substituted as needed; the lipid includes a phospholipid; an amino acid; carbon a water compound; a peptide; or a cholesterol; or another pharmaceutically acceptable leaving group; in one embodiment, the compound is administered as a 5'-phosphonate lipid or a 5'-ether lipid; 12 200835507

Ri及R2各自分別為Η、視需要可經取代之直鏈、分支、或環狀烷基（包括低碳烷基）、烯基或炔基、C〇-烷基、CO_ 烯基、CO-炔基、CO-芳基或雜芳基、C〇-烧氧基烷基、co_ 芳氧基烷基、CO-經取代之芳基、磺醯基、烷基磺醯基、芳 5 基磺醯基、芳烷基磺醯基； (η)如下式V之爾洛堤尼系列化合物：Ri and R2 are each independently a linear, branched, or cyclic alkyl group (including a lower alkyl group), an alkenyl group or an alkynyl group, a C〇-alkyl group, a CO-alkenyl group, or a CO- group which may be substituted as needed. Alkynyl, CO-aryl or heteroaryl, C〇-alkoxyalkyl, co-aryloxyalkyl, CO-substituted aryl, sulfonyl, alkylsulfonyl, aryl 5 sulfonate Mercapto, aralkylsulfonyl; (η) a compound of the following formula V:

式V 其中 *二=2分別為氫、分別視需要可經取代之燒氧基、視而σ、、工代之胺、芳香族胺、雜芳香族胺、視+ 取代之直鏈、分支或環㈣基；"日祕要可經 15Wherein *2 = 2 are respectively hydrogen, respectively, if desired, a substituted alkoxy group, σ, a working amine, an aromatic amine, a heteroaromatic amine, a linear + substituted straight chain, a branch or Ring (four) base; "Day secret to pass 15

族胺:雜芳香族胺或環狀胺;以及視而要了、〜料㈣藥學上可接受之載劑。 =另個貫施例中，本發明涵療腫瘤或癌症之大& |種於南礼動物治種組成物包括： &括對该哺乳動物投予有致量之_ ⑴式I-IV化合物： 13 0¾ 200835507Aminamine: a heteroaromatic amine or a cyclic amine; and, if desired, a pharmaceutically acceptable carrier. In another embodiment, the invention relates to a large tumor of the tumor or cancer, and the composition of the plant of the Nanli animal includes: & comprises administering to the mammal a measurable amount of _ (1) a compound of the formula I-IV : 13 03⁄4 200835507

5 其中R2,、R3’及R5’分另丨J為氫、視需要可經取代之磷酸根或膦酸根（包括一磷酸、二磷酸、或三磷酸或安定化磷酸前藥）；醯基（包括低碳醯基）；烷基（包括低碳烷基）；醯胺、石黃酸S旨包括烧基或芳基烧基；績醯基包括甲續醯基及节基，其中該苯基視需要可經以一個或多個如於此處所述芳 10 基之定義中說明之取代基取代；視需要可經取代之芳基磺醯基；脂質包括磷脂質；胺基酸；碳水化合物；胜肽；或膽固醇；或其它藥學上可接受之離去基，其於活體内可提 14 200835507 供一種化合物其中R2,、R3,或R5,分別為Η或一磷酸、二磷酸、或三磷酸；其中Rx&Ry分別為氫、視需要可經取代之磷酸根；醢基（包括低碳醯基）；醯胺、烷基（包括低碳烷基）；芳香族、 5 去氧伸烷基諸如聚乙二醇、視需要可經取代之芳基磺醯基；脂質包括磷脂質；胺基酸；碳水化合物；胜肽；或膽固醇；或其它藥學上可接受之離去基；於一個實施例中，該化合物係呈5，-磷醚脂質或5，-醚脂質投藥；5 wherein R2, R3' and R5' are further 丨J is hydrogen, optionally substituted phosphate or phosphonate (including monophosphate, diphosphate, or triphosphate or stabilized phosphate prodrug); Including a low carbon fluorenyl group; an alkyl group (including a lower alkyl group); a decylamine, a tartaric acid S is intended to include a decyl group or an aryl group; the fluorenyl group includes a fluorenyl group and a benzyl group, wherein the phenyl group If desired, may be substituted with one or more substituents as described in the definition of the aryl group 10 as described herein; optionally substituted arylsulfonyl; lipids including phospholipids; amino acids; carbohydrates ; peptide; or cholesterol; or other pharmaceutically acceptable leaving group, which can be raised in vivo 14 200835507 for a compound wherein R 2 , R 3 , or R 5 are respectively hydrazine or monophosphate, diphosphate, or tri Phosphoric acid; wherein Rx&Ry are respectively hydrogen, optionally substituted phosphate; sulfhydryl (including lower sulfhydryl); decylamine, alkyl (including lower alkyl); aromatic, 5 deoxyalkylene Base such as polyethylene glycol, optionally substituted arylsulfonyl; lipids including phospholipids; amines Acid; a carbohydrate; peptide; or cholesterol; or other pharmaceutically acceptable leaving group; in one embodiment, the compound is the form of 5 - ether lipid phosphorus or 5, - ether lipid administration;

Ri及R2各自分別為Η、視需要可經取代之直鏈、分支、 10或環狀烷基（包括低碳烷基）、烯基或炔基、CO-烷基、CO-稀基、CO-炔基、C0_芳基或雜芳基、c〇_烷氧基烷基、CO-芳氧基烷基、CO·經取代之芳基、磺醯基、烷基磺醯基、芳基磺醯基、芳烷基磺醯基；以及 (η)如下式V之爾洛堤尼系列化合物：Ri and R2 are each a linear, branched, 10 or cyclic alkyl group (including a lower alkyl group), an alkenyl group or an alkynyl group, a CO-alkyl group, a CO-saturated group, and a CO which may be substituted as needed. - alkynyl, C0_aryl or heteroaryl, c〇_alkoxyalkyl, CO-aryloxyalkyl, CO. substituted aryl, sulfonyl, alkylsulfonyl, aryl a sulfonyl group, an aralkyl sulfonyl group; and (η) a compound of the following formula V:

其中需要可經取代之胺Which requires a substituted amine

Ri及R2分別錢、分別視需要可經取代之聽基、視芳香族胺、雜芳香族胺、視需要可經取代之直鏈、分支或環狀烧基； 4各自刀別為氫、分別為視需要可經取代之芳香矢月女、雜芳香族胺或環狀胺。忒等方法可用於治療對TCN 、TCN-P及/或相關化合物 15 200835507 之毒性效應特別敏感之腫瘤及癌症。於另一個實施例中，提t、於哺乳動物特別力人類治療腫瘤之方法包括⑴由腫瘤獲得生物樣本：⑼判定賴瘤是㈣度表現Akt激酶；以及㈣以如此處所述之曲西立濱、石舞酸曲西立濱或相關化合 5物組5種或多種爾洛堤尼系列化合物來治療過度表現 Akt激酶之腫瘤。於另—個實施例中，經由檢定分析腫瘤或癌症疋否有麟酸化Akt激酶的存在，例如經由使用可檢測磷酸化形式之抗體來判定Akt激酶之表現程度。於另一個實施例中，經由檢定分析得自個體之腫瘤細胞或癌細胞且比較 1〇該含量與對照組織之含量，可判定Akt之過度表現程度。於若干實施例中，於癌症樣本比較對照組，Akt可至少2、2.5、 3、或5倍過度表現。於若干實施例中，過度表現之Akt激酶可為過度活化且鱗酸化之Akt激酶。於本發明之另一個態樣中，提供用藥計劃其可限制 15 TCN及相關化合物之毒性副作用。於另一個實施例中，此種給藥計劃可減少或消除毒性副作用，包括但非限於肝毒性、血小板減少、高血糖、σ區吐、低企約、貧血、低白蛋白血症、骨髓抑制、血中三酸甘油酯過高、血中澱粉酶過咼、腹瀉、胃炎及/或發燒。於另一個實施例中，tcn、Tcn-p 20或相關化合物之投藥可於至少15%、20%或25%個體於活體内提供至少部分諸如至少15%、20%或30%反應或完全反應0 於另一個實施例中，提供一種方法來治療已經診斷為患有腫瘤之個體，係經由對該個體投予有效量之TCN、 16 200835507 TCN-Ρ或相關化合物及一種或多種爾洛堤尼系列化合物，该投樂係根據〆種給藥計劃’包括約略每週一次投予曲西立濱化合物及爾洛堤尼系列化合物約經3週，接著為未投予曲西立濱化合物及爾洛堤尼系列化合物之一週時間。另一 5個實施例中，提供一種於一個體治療腫瘤或癌症之方法，係經由對該個體投予一種給藥計劃，每週各一次投予1〇毫克/平方米或以下之TCN、TCN-P或相關化合物及一種或多種爾洛堤尼系列化合物。於另一個實施例中，曲西立濱化合物及一種或多種爾洛堤尼系列化合物可以短時間例如約 10 5、10或15分鐘時間以單一大劑量投予。於又有其它實施例中，提供給藥計劃，其中曲西立濱化合物及一種或多種爾洛堤尼系列化合物係透過連續輸注經歷至少24、48、72、 96或120小時時間投予。於若干實施例中，連續投藥可每週至少重複一次、每兩週重複一次、及/或每個月重複一次。 15 於其它實施例中，曲西立濱化合物及一種或多種爾洛堤尼系列化合物可每三週至少投予一次。於額外實施例中，化合物可每日至少一次投藥經歷至少2、3、4或5日。於額外實施例中，如此處揭示之曲西立濱化合物及一種或多種爾洛堤尼系列化合物可以有效造成腫瘤退行之數 20 量投予病人。曲西立濱化合物及一種或多種爾洛堤尼系列化合物之投予可於活體内於至少15-20%個體提供至少部分反應，諸如至少15%、20%或30%反應或完全反應。於若干實施例中，至少2、5、10、15、20、30或50毫克/平方米曲西立濱化合物及至少約10、25、50、75、100、150、2〇〇、 17 200835507 5 10 15 20 250或5GG毫克之如此處揭*之—種或多種爾洛堤尼系列化合物可投予一個體。曲西立濱化合物及一種或多種爾洛堤尼系列化合物之投予可根據此處揭示之任一種治療計劃進行。於特殊實施例中，給藥計劃包括投予低於2〇毫克/平方米曲西立濱化合物及一種或多種爾洛堤尼系列化合物。於一個實施例中，每週一次投予低於1〇毫克/平方米曲西立濱化合物及低於約2GG*克之—種或多種_洛堤尼系列化合物。於其它實施例中，低於2毫克/平方米、口 10W⑽请丨嶋及10毫克、20毫克、50毫克、100毫克、15〇毫克、2〇〇毫克、 250亳克、或500毫克劑量之一種或多種爾洛堤尼系列化合物投予-個體。於另-個實施例中，低於1{)毫克/平方米: 西立濱化合物及低於150毫克ϋ洛堤尼化合物可透過連續輸注經歷至少5日投予-個體。於特定實施例中，如此處揭不之曲西立濱化合物及-種或多種爾洛堤尼系列化合物可用於胰癌、攝護腺癌、大腸直腸癌及/或㈣癌之治療。於另-個實施例中，曲西立濱化合物及—種或多種爾洛堤尼系列化合物及/或本發明之治療計劃可用來治療癌瘤、肉瘤、淋巴瘤 '白血病及/或骨髄瘤。於本發明之豆它㈣财’ ㈣及—種或多種爾洛堤尼㈣化合物可餘治療實體腫瘤。又有其它實施例中，此處揭示之曲西立濱化合物及-種或多種爾洛堤尼系列化合物及植成物可用於治顏m症，諸如但雜於下㈣官或组織之癌症：***、攝護腺、硬骨、肺、大腸包括^非限制 18 200835507 大腸直腸、尿路、膀胱、非何杰金氏淋巴瘤、黑素瘤、腎臟、腎上腺、胰臟、咽頭、甲狀腺、胃、腦及/或卵巢。於特定實施例中，曲西立濱化合物及一種或多種爾洛堤尼系列化合物可用於胰癌、乳癌、大腸直腸癌、及/或卵巢癌之 5 治療。於本發明之其它實施例中，此處揭示之曲西立濱化合物及一種或多種爾洛堤尼系列化合物可用於治療血管新生相關疾病。於若干實施例中，提供透過連續輸注歷至少 24、48、72或96小時，經由連續輸注曲西立濱化合物及一種或多種爾洛堤尼系列化合物來治療白血病。於其它實施 10 例中，連續輸注例如可重複每兩週、每三週或每四週至少一次。於特定實施例中，提供於一宿主治療腫瘤癌症及其它與異常細胞增生相關之病症之方法，該方法包括對該宿主投予有效量之曲西立濱化合物及一種或多種爾洛堤尼系列 15 化合物視需要可組合藥學上可接受之載劑。於一個態樣中，曲西立濱化合物及一種或多種爾洛堤尼系列化合物及組成物可組合投予，形成該組成物之一部分，或可同時或不同時呈分開組成物投予。於其它實施例中，如此處揭示之曲西立濱化合物及一 20 種或多種爾洛堤尼系列化合物可用來治療對一種或多種已知抗癌藥物具有抗藥性之腫瘤或癌症，包括此處揭示之腫瘤或癌症及化合物之實施例。於一個實施例中，如此處揭示曲西立濱化合物及一種或多種爾洛堤尼系列化合物係以用於治療患有藥物抗性腫瘤或癌症之病人例如多重抗藥性 19 200835507 腫瘤或癌症之病人之有效量投予，包括單獨對紫杉醇 (taxol)、拉帕黴素(rapamydn)、塔莫西芬(tam〇xifen)、西鉑汀(dsplatin)及/或傑費堤尼(gefitinib)(艾瑞莎（iressa))有抗藥性之腫瘤或癌症。 5 於若干實施例中，提供一種方法包括對有需要之宿主投予有效量之如此處揭示曲西立濱化合物及一種或多種爾洛堤尼系列化合物；或以可有效於宿主治療腫瘤、癌症及其它與異常細胞增生相關之病症之治療有效量之包括曲西立濱化合物及一種或多種爾洛堤尼系列化合物之藥學組成 10 物。於另一個實施例中，提供一種治療腫瘤或癌症之方法，包括將有效量之如此處揭示之化合物或其鹽、異構物、前藥、或酉旨投予有需要之個體，其中該癌症例如為癌瘤、肉瘤淋巴瘤、白血病、或骨髓瘤。該化合物或其鹽、異 15構物、前藥或醋視需要可呈藥學上可接受之組成物:供:、組成物包括適當載劑諸如水其係調配用於投予有需要之病人之適當投藥途徑使用。任選地，該化合物可組合至少一種腫瘤或癌症之額外治療劑投予，或與該額外治療劑交替投予。 2〇纟發明也包括如此處揭示之化合物或其鹽、前藥或酉旨用於治療腫瘤或癌症之用途，視需要可於藥學上可接受^ 載劑，以及如此處揭示之曲西立濱化合物及一種或多種爾 CT疋匕系歹〗化合物或其鹽、前藥或酯視需要可於藥學上可接又之載^，用於製造癌症或腫瘤治療用藥之用途。σ 20 200835507 圖式簡單說明第1圖顯示由NCI分集集合中識別作為Akt抑制劑候選者之API-2 (曲西立濱）之識別。第1A圖顯示API_2 (曲西立濱）之化學結構式。第1B圖顯示API-2抑制於AKT2轉形NIH3T3 5 細胞中之AKT2磷酸化程度。威爾（Wile)型AKT2-轉形 NIH3T3細胞以API-2 (1 μΜ)處理指示之時間，且接受以抗石粦酸-Akt-T308及-S473抗體(上圖及中圖）之免疫墨點分析。下圖顯示總AKT2之表現。於第1C圖中，顯示API-2抑制Akt 之三個同質異形體。HEK293細胞於EGF刺激前以 10 HA-Akt 卜 HA-AKT2、及 HA-AKT3 轉移感染且以 API-2 (1 μΜ) 或瓦特曼寧（wortmannin) (15μΜ)處理，細胞經溶解且以 anti-HA抗體免疫沉澱。免疫沉澱物接受試管内激酶檢定分析（上)及以抗磷酸-Akt-T308(下)抗體進行免疫墨點分析。中圖顯示經轉移感染之Aktl、AKT2、及AKT3之表現。第1D 15圖顯示API-2於試管内不會抑制Akt。於試管内組成活性 AKT2重組蛋白質於激酶緩衝液之激酶檢定分析含有1μΜ API-2 (線道3) 〇第2圖驗證API-2不會抑制PI3K、PDK1及AGC激酶家族中之緊密相關成員。第2A圖顯示試管内pi3K激酶檢定分 20析。於EGF刺激前，HEK293細胞經血清匱乏且以API-2 (ΙμΜ)或瓦特曼寧（15μΜ)處理30分鐘。細胞經溶解且以 anti-pllOa抗體免疫沉澱。免疫沉澱物使用ΡΙ_4-Ρ作為酶基質接受活體内激酶檢定分析。第2Β圖顯示ΑΡΙ-2對試管内 PDK1活化之效應（上圖），實心圓顯示藉Αρι_2抑制。空心圓 21 200835507 顯不藉陽性對照史妥洛史寶靈(staurosporine)抑制，史妥洛史寶靈為強力PDK1抑制劑(IC50=5 nM)。下圖為HEK293細胞之免疫墨點分析，HEK293細胞於EGF刺激前以 Myc-；PDK1轉移感染且以瓦特曼寧或API_2處理。免疫墨點 5 係以指示之抗體檢測。第2C圖顯示PKCa以抗磷酸-PKCa •T638(上圖）及總PKCa (下圖）抗體接著以API-2或非選擇性 PKC抑制劑r〇31-8220處理之磷酸化程度之免疫墨點分析。第2D圖顯示試管内Sgk激酶檢定分析。HEK293細胞於 EGF刺激前以HA-SGK轉移感染且以API-2或瓦特曼寧處 10 理。試管試驗之激酶係以HA-SGK免疫沉澱使用MBP作為酶基質進行(上圖）。下圖顯示經轉移感染之HA-SGK之表現。第2E圖顯示PKA激酶檢定分析之結果。經過免疫純化之 PKA於含有適用之抑制劑（API-2或PKAI)及酶基質坎普泰 (Kemptide)之ADB緩衝液（上態生技公司（Upstate 15 Biotechnology Inc))中培養。定量激酶活性。於第2F圖中顯示西方墨點。OVCAR3細胞以API-2處理指示時間。細胞溶解產物係以適用之抗磷酸抗體G-4圖）及抗肌動蛋白抗體 (下圖）進行免疫墨點。第3圖驗證API-2於有升高的Akt之人癌細胞中，抑制 20 Akt活性及細胞生長而誘導細胞凋亡。第3A圖為使用API-2 處理後之西方墨點，Akt之磷酸化程度係以抗磷酸 -Akt-T308抗體於適用之人癌細胞系檢測。墨點再度使用抗總Akt抗體探測（下圖）。第3B圖中，顯示細胞增生檢定分析。圖中指示之細胞系以不同劑量之API_2處理24小時及48 22 200835507 小時，然後以細胞力價（CellTiter) 96細胞增生檢定分析套件組（普米嘉公司（Promega))分析。第3C圖提供細胞凋亡分析。細胞以API-2處理，以附著素(annexin) V及PI染色，及藉FACScan分析。 5 第4圖顯示於小鼠異種移植片中，於有升高之Akt之癌細胞系中，API-2抑制Akt之下游標乾，且具有抗腫瘤活性。於第4A圖中，驗證API-2可抑制結節素(tuberin)、Bad、AFX 及GSK-3/3之Akt石舞酸化。於以API-2處理後，OVCAR3細胞經溶解且以適用之抗體進行免疫墨點分析。第4B圖顯示 10 API-2可抑制腫瘤生長。腫瘤細胞注射入裸小鼠體内，左側 Akt細胞濃度低，而右側Akt細胞濃度高。當腫瘤達到約 100-150立方毫米平均大小時，動物以載媒劑或以1毫克/千克/日API-2處理。各個測量值表示1〇個腫瘤之平均值。第4C 圖顯示帶有以API-2或載媒劑（對照組)處理之QVCAR3 (右） 15及0VCAR5 (左）之小鼠之代表圖。第4D圖顯示於實驗結束時之腫瘤大小（下）及重量（上)之實例。第4E圖中，腫瘤溶解產物之免疫墨點分析係使用經以API-2處理（T3及T4)及未經處理（T1及T2)之〇VCAR_3-衍生腫瘤中之抗石粦_Akt-S473 (上)及抗-AKT2 (下)抗體進行。 20 第5圖顯示API-2 (曲西立濱）抑制於試管内抑制Akt激酶活性。試管内激酶檢定分析係使用之重組株於含磷酸基肌糖醇-3,4,5-P3 (PIP3)、API-2及組織腺H2B作為酶基質之激酶緩衝液進行。培養30分鐘後，藉SDS-PAGE 分離反應且暴露於薄膜。 23 200835507 第6a-6c圖提供人Aktl之mRNA及胺基酸序列，也標示限剪酶位置。第7a-7d圖提供人Akt2之mRNA及胺基酸序列，也標示限剪酶位置。第8a-8c圖提供人Akt3之mRNA及胺基酸序列，也標示限剪酶位置。第9圖顯示藉崔茲竹美(trastuzmnab)及Akt/mTOR徑路抑制劑之組合物之生長抑制。PTEN反訊息或非特異性寡核苷酸轉移感染之BT474.ml細胞單獨以Akt/mTOR徑路抑制 10 劑處理，或以與崔茲竹美之組合物處理，及評估相對細胞生長。第9A圖顯示Akt/mTOR抑制劑之示圖。生長抑制係於經PTEN AS轉移感染之BT474.ml細胞中評估。顯示劑量為：曲西立濱（TCN) ΙμΜ ; RAD001 0.2 nM;QLT〇267 ΙΟμΜ ; KP 372-1 〇·〇5μΜ ; 4ADPIB 5μΜ ;艾朵弗新 15 20 (Edelfosine) 7·5μΜ ;及崔茲竹美(Ttzm) 2微克/毫升。指示標準差（SD)，以生長抑制百分比表示。所示結果為由2 3次實驗所得之組合資料，各實驗中之各項處理重複三次。第 9B圖顯示TCN與崔茲竹美組合抑制細胞生長。BT474❿^細胞以PTEN AS寡核苷酸或非特異性(NS)募核苷酸轉移感染，以崔茲竹美及TCN於多劑TCN單獨處理或組合處理，且檢定分析生長抑制。崔茲竹美係以單一濃度投予。第9C 圖顯示RAD001與崔茲竹美組合抑制細胞生長。1 細胞以PTEN AS募核苷酸或非特異性(NS)寡核苷染，以崔茲竹美及RAD001於多劑RAD001單獨處酸轉移感理或組合 24 200835507 處理，且檢定分析生長抑制。用於第9B及9C圖：*指示比較單獨使用崔茲竹美或TCN/TAD001，於組合處理後生長抑制之顯著差異。P<0.05被視為顯著。誤差柱顯示SEM。第10圖顯示用於細胞凋亡之協同效果。於接種後24小 5 時，經PTEN AS及NS轉移感染之BT474.ml細胞如所指示以於下列濃度之崔茲竹美(Ttzm)、TCN及/或RAD001處理··崔茲竹美2微克/毫升；曲西立濱2·5μΜ ; RAD001 0.4 nM。進行Apo-BrdU土諾(Tunel)檢定分析來評估細胞凋亡。實驗進行三次，所示資料為平均細胞凋亡。誤差柱顯示標準差。 10 比較全部其它處理，崔茲竹美+曲西立濱處理顯著誘導細胞凋亡（ρ<0·01)。第11圖顯示Akt及P70S6K活性之抑制。為了評估此等藥物用於Akt/mTOR徑路之效果，PTEN AS及NS寡核苷酸係於 BT474.ml細胞轉移感染。二日後，細胞以崔茲竹美及曲西 15 立濱(TCN)(第11A圖）或崔茲竹美及RAD001(第11B圖）處理 2小時。收集全細胞溶解產物，藉SDS-PAGE分離及如指示進行免疫墨點分析。崔茲竹美之濃度為2微克/毫升，曲西立濱為2·5μΜ及RAD001為〇.4nM。實驗至少重複兩次來確保結果為可再現性。 2〇第12圖顯示於SCID小鼠異種移植片模型中組合治療抑制腫瘤生長。SCID小鼠接受BT474.ml乳癌細胞異種移植片於乳腺脂肪墊。異種移植片成長3週來產生平均大小 100-150立方毫米之腫瘤。投予pTEN反訊息募核苷酸、崔茲竹美、曲西立濱（第12A圖）及RAD001(第12B圖）。使用測 25 200835507 微器每週兩次測量腫瘤大小，對各處理組求取腫瘤大小平均。誤差柱表示平均值之標準差。*指示比較單獨崔茲竹美 (Ttzm)、TCN或DMSO，組合治療後之生長抑制有顯著差異。P<0.05被視為顯著。 5 【實施方式】較佳實施例之詳細說明與先前技術及經驗相反，發明人已經判定如何成功地使用曲西立濱化合物組合一種或多種爾洛堤尼系列化合物藉以下一種方式或其組合來治療腫瘤及癌症：⑴將曲西立 10 濱及一種或多種爾洛堤尼系列化合物只投予根據後述診斷試驗對曲西立濱化合物及/或爾洛堤尼系列化合物之敏感度升高之病人；（ii)使用規定之劑量水平，其可最小化曲西立濱化合物及/或爾洛堤尼系列化合物之毒性，同時仍然具有功效；或(iii)使用所述用藥計劃，其可最小化曲西立濱化 15 合物及/或爾洛堤尼系列化合物之毒性。 5.1定義如此處使用，「本發明化合物」一詞係指此處揭示之化合物包括式I-VIII化合物及其組合。如此處使用，「癌症」及/或「癌性」等詞述及或說明 20 哺乳動物體内典型以未經調節之細胞生長為特徵之生理症狀，亦即增生病症。此種增生病症之實例包括癌症諸如癌瘤、淋巴瘤、母細胞瘤、肉瘤及白血病，以及此處揭示之其它癌症。此等癌症之更特定實例包括乳癌、攝護腺癌、大腸癌、鱗狀細胞癌、小細胞肺癌、非小細胞肺癌、胃腸 26 200835507 癌、胰癌、子宮頸癌、卵巢癌、肝癌例如肝癌瘤、膀胱癌、大腸直腸癌、子宮内膜癌、腎癌及曱狀腺癌。其它癌症之非限制性實例為基底細胞癌；膽管癌；骨癌；腦及中樞神經系統(CNS)癌；脈絡膜癌；結締組織癌； 5食運癌；眼癌；頭頸癌；胃癌；上皮内腫瘤；咽癌；淋巴 ‘匕括何杰金氏淋巴瘤及非何杰金氏淋巴瘤；黑素瘤；骨知瘤，神纟1母細胞瘤，口腔癌（例如唇、舌、口及喉頭）；姨癌，視網膜母細胞瘤；橫紋肌肉瘤；直腸癌；呼吸系統癌；肉瘤，皮膚癌；胃癌；睪丸癌；子宮癌；泌尿系統癌及其 10它癌瘤及肉瘤。、，如此處使用，「腫瘤」一詞係指全部贅生性細胞生長及匕括惡性或良性及全部癌前及癌性細胞及組織。例 σ特定癌症可以實體團塊腫瘤為特徵。實體腫癌闈塊當Ri and R2 respectively, respectively, which can be substituted, such as an amine group, an aromatic amine, a heteroaromatic amine, or a linear, branched or cyclic alkyl group which may be substituted as required; An aromatic sagittal, heteroaromatic or cyclic amine which may be substituted as needed. Methods such as sputum can be used to treat tumors and cancers that are particularly sensitive to the toxic effects of TCN, TCN-P, and/or related compound 15 200835507. In another embodiment, the method for treating a tumor in a mammal, particularly for humans, comprises (1) obtaining a biological sample from the tumor: (9) determining that the tumor is a (four) degree of Akt kinase; and (iv) using Quxili as described herein. 5 or more of the erlotinib compounds of the sulphate, the sirolimus or the related compound 5 group to treat tumors that overexpress Akt kinase. In another embodiment, the presence of a sulphated Akt kinase is analyzed by assay for tumor or cancer, e.g., by using antibodies that detect the phosphorylated form to determine the extent of Akt kinase expression. In another embodiment, the degree of overexpression of Akt can be determined by assaying tumor cells or cancer cells obtained from an individual and comparing the amount of the tumor cells to the control tissue. In several embodiments, Akt can be overexpressed at least 2, 2.5, 3, or 5 fold in a control sample compared to a cancer sample. In several embodiments, the overexpressed Akt kinase can be an overactivated and squamized Akt kinase. In another aspect of the invention, a medication regimen is provided which limits the toxic side effects of 15 TCN and related compounds. In another embodiment, such a dosing schedule reduces or eliminates toxic side effects including, but not limited to, hepatotoxicity, thrombocytopenia, hyperglycemia, sigma sputum, hypovolemia, anemia, hypoalbuminemia, myelosuppression , high triglyceride in the blood, amylase in the blood, diarrhea, gastritis and / or fever. In another embodiment, the administration of tcn, Tcn-p 20 or a related compound provides at least a portion, such as at least 15%, 20% or 30%, of the reaction or complete response in at least 15%, 20% or 25% of the individual in vivo. In another embodiment, a method is provided for treating an individual who has been diagnosed with a tumor by administering to the individual an effective amount of TCN, 16 200835507 TCN-Ρ or related compound and one or more of the Lartini series The compound, according to the sputum dosing plan, includes about once a week of administration of the triclinide compound and the erlotini series of compounds for about 3 weeks, followed by the failure to administer the triclinide compound and erlo One of the Tenny series compounds in the week. In another five embodiments, a method for treating a tumor or a cancer in a body is provided, by administering a dose to the individual, and administering TCN, TCN at a dose of 1 mg/m 2 or less per week. -P or related compounds and one or more erlotini series compounds. In another embodiment, the triclinide compound and one or more erlotinib compounds can be administered in a single large dose over a short period of time, e.g., about 105, 10, or 15 minutes. In still other embodiments, a dosing schedule is provided wherein the triplicidin compound and one or more ritotinic series compounds are administered by continuous infusion for a period of at least 24, 48, 72, 96 or 120 hours. In several embodiments, continuous administration can be repeated at least once a week, repeated every two weeks, and/or repeated once a month. In other embodiments, the triclinide compound and one or more erlotinib compounds can be administered at least once every three weeks. In additional embodiments, the compound can be administered at least once a day for at least 2, 3, 4 or 5 days. In additional embodiments, the triclinide compound and one or more of the erlotinib compounds as disclosed herein are effective to cause tumor regression to be administered to a patient. Administration of a triclinide compound and one or more erlotinib compounds can provide at least a partial response, such as at least 15%, 20% or 30%, of the reaction or complete reaction in at least 15-20% of the individual in vivo. In some embodiments, at least 2, 5, 10, 15, 20, 30 or 50 mg/m 2 of triclinide compound and at least about 10, 25, 50, 75, 100, 150, 2, 17 200835507 5 10 15 20 250 or 5 GG mg of one or more of the erlotini series compounds can be administered to one body as disclosed herein. Administration of the triclinide compound and one or more of the erlotini series compounds can be carried out according to any of the treatment plans disclosed herein. In a particular embodiment, the dosing schedule comprises administering less than 2 mg/m2 of the triclinide compound and one or more of the Lerotines series of compounds. In one embodiment, less than 1 mg/m2 of tricineridine compound and less than about 2 GG*g of one or more _Lotini series compounds are administered once a week. In other embodiments, less than 2 mg/m 2 , mouth 10 W (10), and 10 mg, 20 mg, 50 mg, 100 mg, 15 mg, 2 mg, 250 g, or 500 mg dose One or more of the erlotinib compounds are administered to the individual. In another embodiment, less than 1 {) mg/m 2 : the cililibine compound and less than 150 mg of the zenotinic compound can be administered to the individual via continuous infusion for at least 5 days. In a particular embodiment, the triclinide compound and one or more of the erlotinib compounds as disclosed herein can be used in the treatment of pancreatic cancer, prostate cancer, colorectal cancer, and/or (four) cancer. In yet another embodiment, the triclinide compound and one or more of the erlotini series compounds and/or the treatment plan of the invention can be used to treat cancer, sarcoma, lymphoma 'leukemia and/or osteosarcoma. In the present invention, the bean (four) financial (four) and one or more of the erlotini (d) compounds can be used to treat solid tumors. In still other embodiments, the tromethamine compound disclosed herein and the one or more of the erlotini series of compounds and plantings can be used to treat cancer, such as but mixed with the lower (four) official or tissue cancer. : Breast, prostate, hard bone, lung, large intestine including ^ non-restricted 18 200835507 Large intestine rectum, urinary tract, bladder, non-Hodgkin's lymphoma, melanoma, kidney, adrenal gland, pancreas, pharynx, thyroid, stomach , brain and / or ovary. In a particular embodiment, the triclinide compound and one or more erlotinilin compounds are useful in the treatment of pancreatic cancer, breast cancer, colorectal cancer, and/or ovarian cancer. In other embodiments of the invention, the triclinide compound disclosed herein and one or more of the erlotinib compounds are useful in the treatment of vascular neonatal related diseases. In several embodiments, the treatment of leukemia is provided by continuous infusion of the triclinide compound and one or more erlotinib series compounds by continuous infusion for at least 24, 48, 72 or 96 hours. In other embodiments, continuous infusion can be repeated, for example, every two weeks, every three weeks, or at least once every four weeks. In a specific embodiment, there is provided a method of treating a tumor cancer and other disorders associated with abnormal cell proliferation in a host, the method comprising administering to the host an effective amount of a triplicidin compound and one or more erlotini series 15 The compound may be combined with a pharmaceutically acceptable carrier as needed. In one aspect, the triclinide compound and one or more erlotini series compounds and compositions can be administered in combination to form part of the composition, or can be administered as separate compositions at the same time or at different times. In other embodiments, a tripecline compound as disclosed herein and a 20 or more erlotinib compounds can be used to treat a tumor or cancer resistant to one or more known anticancer drugs, including Examples of tumors or cancers and compounds are disclosed. In one embodiment, a triclinide compound and one or more erlotinib compounds are disclosed herein for use in treating a patient having a drug resistant tumor or cancer, such as a multidrug resistant 19 200835507 tumor or cancer patient An effective amount of administration, including taxol alone, rapamydn, tam〇xifen, dsplatin, and/or gefitinib (ai Ressa (iressa) has a drug-resistant tumor or cancer. In some embodiments, a method is provided comprising administering to a host in need thereof an effective amount of a triclinide compound and one or more erlotinib compounds as disclosed herein; or in treating a tumor, cancer And a therapeutically effective amount of other disorders associated with abnormal cell proliferation comprising a citrilide compound and one or more pharmaceutical compositions of the erlotinib series of compounds. In another embodiment, a method of treating a tumor or cancer comprising administering an effective amount of a compound as disclosed herein, or a salt, isomer, prodrug, or hydrazine thereof, to a subject in need thereof, wherein the cancer is For example, cancer, sarcoma lymphoma, leukemia, or myeloma. The compound or a salt thereof, iso- 15th structure, prodrug or vinegar may optionally be in the form of a pharmaceutically acceptable composition: for: a composition comprising a suitable carrier such as water for its administration to a patient in need thereof Use the appropriate route of administration. Optionally, the compound can be administered in combination with or in addition to an additional therapeutic agent for at least one tumor or cancer. The invention also includes the use of a compound as disclosed herein, or a salt, prodrug or prodrug thereof, for the treatment of a tumor or cancer, a pharmaceutically acceptable carrier, if desired, and triclinide as disclosed herein. The compound and one or more of the compounds of the CT system, or a salt, prodrug or ester thereof, may be pharmaceutically acceptable, and may be used for the manufacture of a cancer or a tumor therapeutic drug. σ 20 200835507 Schematic description of the figure Figure 1 shows the identification of API-2 (Quxi Libin) identified as a candidate for Akt inhibitors in the NCI diversity set. Figure 1A shows the chemical structure of API_2 (Quxi Libin). Figure 1B shows the extent of AKT2 phosphorylation in API-2 inhibited by AKT2 transformed NIH3T3 5 cells. Will-type AKT2-transformed NIH3T3 cells were treated with API-2 (1 μΜ) for the indicated time and received immunostaining with anti-phosphinic acid-Akt-T308 and -S473 antibodies (top and bottom) Point analysis. The figure below shows the performance of the total AKT2. In Figure 1C, three isomorphs of API-2 inhibition of Akt are shown. HEK293 cells were infected with 10 HA-Akt, HA-AKT2, and HA-AKT3 prior to EGF stimulation and treated with API-2 (1 μΜ) or wortmannin (15 μΜ). The cells were lysed and anti-anti- HA antibody immunoprecipitation. The immunoprecipitate was subjected to in-vitro kinase assay (top) and immunoblot analysis with anti-phospho-Akt-T308 (bottom) antibody. The middle panel shows the performance of Aktl, AKT2, and AKT3 by transfer infection. Figure 1D 15 shows that API-2 does not inhibit Akt in the test tube. The kinase assay for the active AKT2 recombinant protein in a kinase buffer containing 1 μΜ API-2 (line 3) 〇 Figure 2 demonstrates that API-2 does not inhibit closely related members of the PI3K, PDK1 and AGC kinase families. Figure 2A shows the pi3K kinase assay in vitro. HEK293 cells were serum-deficient and treated with API-2 (ΙμΜ) or Watmanning (15 μΜ) for 30 minutes prior to EGF stimulation. The cells were lysed and immunoprecipitated with an anti-pllOa antibody. The immunoprecipitate was assayed for in vivo kinase assay using ΡΙ_4-Ρ as the enzyme substrate. Figure 2 shows the effect of ΑΡΙ-2 on the activation of PDK1 in the test tube (above), and the solid circle shows inhibition by Αρι_2. Hollow rounds 21 200835507 were not inhibited by the positive control staurosporine, and Stromoxol was a potent PDK1 inhibitor (IC50 = 5 nM). The following figure shows the immunoblotting analysis of HEK293 cells. HEK293 cells were infected with Myc-; PDK1 prior to EGF stimulation and treated with Watmanin or API_2. The immune dot 5 is detected by the indicated antibody. Figure 2C shows immunofluorescence of PKCa with phosphorylation to anti-phospho-PKCa • T638 (top panel) and total PKCa (bottom panel) antibodies followed by API-2 or non-selective PKC inhibitor r〇31-8220 analysis. Figure 2D shows an in vitro Sgk kinase assay. HEK293 cells were infected with HA-SGK transfer prior to EGF stimulation and treated with API-2 or Watmanin. The test tube kinase was immunoprecipitated with HA-SGK using MBP as the enzyme substrate (top panel). The figure below shows the performance of HA-SGK transfected with infection. Figure 2E shows the results of the PKA kinase assay. The immunopurified PKA was cultured in ADB buffer (Upstate 15 Biotechnology Inc) containing the appropriate inhibitor (API-2 or PKAI) and the enzyme substrate Kemptide. Quantify kinase activity. Western blots are shown in Figure 2F. OVCAR3 cells were timed with API-2 treatment. The cell lysate was immunized with the anti-phospho antibody G-4 map and the anti-actin antibody (bottom panel). Figure 3 demonstrates that API-2 induces apoptosis by inhibiting 20 Akt activity and cell growth in human cancer cells with elevated Akt. Figure 3A shows Western blots treated with API-2. The degree of phosphorylation of Akt is detected by anti-phospho-Akt-T308 antibody in a suitable human cancer cell line. The ink spots were again probed with anti-total Akt antibody (bottom panel). In Fig. 3B, a cell proliferation assay is shown. The cell lines indicated in the figure were treated with different doses of API_2 for 24 hours and 48 22 200835507 hours, and then analyzed by the CellTiter 96 Cell Proliferation Assay Kit (Promega). Figure 3C provides an analysis of apoptosis. Cells were treated with API-2, stained with annexin V and PI, and analyzed by FACScan. 5 Figure 4 shows that in mouse xenografts, API-2 inhibits the downstream of Akt in cancer cell lines with elevated Akt and has antitumor activity. In Figure 4A, validation of API-2 inhibited acidification of Akt stone dance of tuberin, Bad, AFX and GSK-3/3. After treatment with API-2, OVCAR3 cells were lysed and subjected to immunoblot analysis with applicable antibodies. Figure 4B shows that 10 API-2 inhibits tumor growth. Tumor cells were injected into nude mice with a low concentration of Akt cells on the left and a high concentration of Akt cells on the right. When the tumor reached an average size of about 100-150 cubic millimeters, the animals were treated with vehicle or at 1 mg/kg/day API-2. Each measurement represents the average of 1 tumor. Figure 4C shows a representation of mice with QVCAR3 (right) 15 and 0VCAR5 (left) treated with API-2 or vehicle (control). Figure 4D shows an example of tumor size (bottom) and weight (top) at the end of the experiment. In Fig. 4E, the immunoblot analysis of tumor lysates was performed using anti-shik _Akt-S473 in VCAR_3-derived tumors treated with API-2 (T3 and T4) and untreated (T1 and T2). (top) and anti-AKT2 (bottom) antibodies were performed. Figure 5 shows that API-2 (Cucidine) inhibits Akt kinase activity in vitro. The in vitro PCR assay was performed using a recombinant strain containing kinases containing phospho-inositol-3,4,5-P3 (PIP3), API-2 and tissue gland H2B as an enzyme substrate. After 30 minutes of incubation, the reaction was separated by SDS-PAGE and exposed to the membrane. 23 200835507 Figure 6a-6c provides the mRNA and amino acid sequence of human Aktl, also indicating the position of the restriction enzyme. Figures 7a-7d provide mRNA and amino acid sequences of human Akt2, also indicating the position of the restriction enzyme. Figures 8a-8c provide mRNA and amino acid sequences of human Akt3, also indicating the position of the restriction enzyme. Fig. 9 shows growth inhibition by a composition of trastuzmnab and an Akt/mTOR pathway inhibitor. BT474.ml cells infected with PTEN anti-message or non-specific oligonucleotides were treated with Akt/mTOR pathway inhibition alone for 10 doses, or treated with a composition of Tries, and evaluated for relative cell growth. Figure 9A shows a diagram of an Akt/mTOR inhibitor. Growth inhibition was assessed in BT474.ml cells infected with PTEN AS transfer. The displayed doses were: triclinide (TCN) ΙμΜ; RAD001 0.2 nM; QLT〇267 ΙΟμΜ; KP 372-1 〇·〇5μΜ; 4ADPIB 5μΜ; Ai Dufuxin 15 20 (Edelfosine) 7·5μΜ; Bamboo (Ttzm) 2 μg / ml. Indicates the standard deviation (SD), expressed as a percentage of growth inhibition. The results shown are the combined data obtained from 23 experiments, and each treatment in each experiment was repeated three times. Figure 9B shows that TCN combined with Tries Zhumei inhibits cell growth. BT474❿ cells were transfected with PTEN AS oligonucleotides or non-specific (NS) nucleotides, treated with Trizin and TCN in multiple doses of TCN alone or in combination, and assayed for growth inhibition. Triz Bamboo is administered at a single concentration. Figure 9C shows that RAD001 combined with Tries Zhumei inhibits cell growth. 1 Cells were stained with PTEN AS-raised nucleotides or non-specific (NS) oligonucleosides, treated with CuiZhumei and RAD001 in multiple doses of RAD001 alone or with a combination of 24 200835507, and assayed for growth inhibition. For Figures 9B and 9C: * indicates a significant difference in growth inhibition after combination treatment, using Trizut or TCN/TAD001 alone. P < 0.05 was considered significant. The error bars show SEM. Figure 10 shows the synergistic effect for apoptosis. At 24 hours after inoculation, BT474.ml cells infected with PTEN AS and NS were treated as follows at the following concentrations of Ttzm, TCN and/or RAD001. /ml; Quxi Libin 2·5μΜ; RAD001 0.4 nM. Apo-BrdU Tunel assay was performed to assess apoptosis. The experiment was performed three times and the data shown were mean apoptosis. The error bars show the standard deviation. 10 Comparing all other treatments, Cui Zhemei + Quxi Libin treated significantly induced apoptosis (ρ < 0·01). Figure 11 shows inhibition of Akt and P70S6K activity. To assess the effect of these drugs on the Akt/mTOR pathway, PTEN AS and NS oligonucleotides were infected with BT474.ml cells. Two days later, the cells were treated for 2 hours with Cui Zhu Zhumei and Quxi 15 Libin (TCN) (Fig. 11A) or Cui Zhumei and RAD001 (Fig. 11B). Whole cell lysates were collected, separated by SDS-PAGE and subjected to immunoblot analysis as indicated. The concentration of Cuiz Bamboo is 2 μg/ml, Quxi Libin is 2·5 μΜ and RAD001 is 〇.4 nM. The experiment is repeated at least twice to ensure that the result is reproducible. 2〇 Figure 12 shows that combination therapy inhibits tumor growth in a SCID mouse xenograft model. SCID mice received BT474.ml breast cancer xenografts on mammary fat pads. Xenografts were grown for 3 weeks to produce tumors with an average size of 100-150 mm3. PTEN anti-information nucleotides, Cui Zhemei, Quxi Libin (Fig. 12A) and RAD001 (Fig. 12B) were administered. Tumor size was measured twice a week using the 2008 35507 microarray, and tumor size was averaged for each treatment group. The error bars represent the standard deviation of the mean. *Indicating a comparison of Ttzm, TCN or DMSO alone, there was a significant difference in growth inhibition after combination therapy. P < 0.05 was considered significant. 5 DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The detailed description of the preferred embodiments is contrary to the prior art and experience, and the inventors have determined how to successfully use the triclinide compound in combination with one or more of the erlotini series compounds in one of the following ways or a combination thereof. Treatment of tumors and cancer: (1) The addition of tromethamine 10 and one or more of the erlotini series compounds is only increased according to the diagnostic test described below for the sensitivity of the triclinide compound and/or the erlotini series of compounds. a patient; (ii) using a prescribed dosage level that minimizes the toxicity of the tricineribine compound and/or the erlotinib series compound while still having efficacy; or (iii) using the medication regimen, which is minimal Toxicity of sirolimus and/or erlotinib compounds. 5.1 Definitions As used herein, the term "compounds of the invention" means that the compounds disclosed herein include compounds of formula I-VIII and combinations thereof. As used herein, the terms "cancer" and/or "cancerous" describe or describe a physiological condition characterized by unregulated cell growth in a mammal, i.e., a proliferative disorder. Examples of such proliferative disorders include cancers such as carcinomas, lymphomas, blastomas, sarcomas, and leukemias, as well as other cancers disclosed herein. More specific examples of such cancers include breast cancer, prostate cancer, colorectal cancer, squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, gastrointestinal 26 200835507 cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer such as liver cancer Tumor, bladder cancer, colorectal cancer, endometrial cancer, kidney cancer and squamous cell carcinoma. Non-limiting examples of other cancers are basal cell carcinoma; cholangiocarcinoma; bone cancer; brain and central nervous system (CNS) cancer; choroidal carcinoma; connective tissue cancer; 5 food cancer; eye cancer; head and neck cancer; gastric cancer; intraepithelial Tumor; pharyngeal carcinoma; lymphatic sputum including Hodgkin's lymphoma and non-Hodgkin's lymphoma; melanoma; osteoma, scorpion 1 blastoma, oral cancer (eg lip, tongue, mouth and throat) Carcinoma, retinoblastoma; rhabdomyosarcoma; rectal cancer; respiratory cancer; sarcoma, skin cancer; gastric cancer; testicular cancer; uterine cancer; urinary system cancer and its 10 cancer and sarcoma. As used herein, the term "tumor" refers to all neoplastic cell growth and includes malignant or benign and all precancerous and cancerous cells and tissues. Example σ-specific cancers can be characterized by solid mass tumors. Solid tumor

。使用後述技術較為常見於早期檢勺分子分析及表現型分析通常可驗 L的癌症，或病灶是否係來自於另瘤”、、去觸診，只能經由醫〜)或藉針刺抽吸來檢測。. The use of the techniques described later is more common in early detection of molecular analysis and phenotypic analysis of L-cancers, or whether the lesions are from another tumor, to palpation, only through the doctor ~) or by acupuncture suction Detection.

-個部伋的癌症轉移。如此處使用，除丨，除非另行指示，否則烷基一詞包括例如 27 200835507 為Cl至C24之直鏈、分支或環狀第一烴、第二烴、或第三烴，特別包括甲基、三氟甲基、乙基、丙基、異丙基、環丙基、丁基、異丁基、第三丁基、戊基、環戊基、異戊基、新戊基、己基、異己基、環己基、環己基甲基、3-甲基戊基、 5 2,2-一甲基丁基、及2,3-二甲基丁基。烧基視需要例如可經以一個或多個取代基取代，取代基諸如鹵原子(F、C1、Br 或 I)(例如 CF3、2-Br·乙基、CH2F、CH2Q、CH2CF3、或 CF2CF3)、羥基（例如CH2OH)、胺基（例如CH2NH2、 CH2NHCH3、或CH2N(CH3)2)、烷基胺基、芳基胺基、烷氧 10基、芳氧基、峭基、疊氮基(例如CH2N3)、氰基(例如ch2CN)、磺酸、硫酸根、膦酸、磷酸根、或膦酸根，或為未經保護，或如熟諳技藝人士已知可視需要經保護，例如如Greene等人，有機合成保護基，1991年，第2版，約翰威利父子公司，紐約之教示，以引用方式併入此處。 15 除非另行陳明，否則如此處使用，低碳烷基一詞係指 (^至匕飽和直鏈、分支鏈、或若屬適當為環狀(例如環丙基）烷基，包括經取代形式及未經取代形式。烧基胺基或芳基胺基等詞包括分別有—個或兩個烧基或芳基取代基之胺基。 2〇胺基酸一詞包括天然及合成《、yS、胺基酸，包括但非限於蛋白質中出現之胺基酸，亦即甘胺酸、丙胺酸、顯胺酸' 白胺酸、異白胺酸、蛋胺酸、苯基丙胺酸、色胺酸、脯胺酸、絲胺酸、蘇胺酸、半脱胺酸、_酸、天冬醯胺、麩胺、天冬酸、麵胺酸、離胺酸、精胺酸、及 28 200835507 組胺酸。於較佳實施例中，胺基酸為L-組態。另外，胺基酸 < 為丙如:酿基、顯胺驢基、白胺醯基、異白胺醯基、脯胺醢基、笨基丙胺醯基、色胺醯基、蛋胺醯基、甘胺醯基、絲胺酿基、蘇胺醯基、半胱胺醯基、酪胺醯基、天冬醯胺 5 基、糙胺基、天冬醯基、戊二醯基、離胺醯基、精胺醯基、組胺酿基、/3-丙胺醯基、綠胺醯基、点-白胺醯基、0-異白胺醯基、/3-脯胺醯基、/9-苯基丙胺醯基、色胺醯基、/5-蛋胺醯基、甘胺醯基、万_絲胺醯基、石-蘇胺醯基、/5-半胱胺醯基、酪胺醯基、天冬醯胺基、万_麩 10胺基、万―天冬醯基、/5-戊二醯基、/3-離胺醯基、/5-精胺醢基、或点-組胺醯基之衍生物。當使用胺基酸一詞時，被視為天然胺基酸或合成胺基酸酯之各自特定獨立揭示，包栝但#限於呈D組態及L組態之α、召、^或占甘胺酸、丙胺酸、纈胺酸、白胺酸、異白胺酸、蛋胺酸、苯基丙胺酸、 15色胺酸、脯胺酸、絲胺酸、蘇胺酸、半胱胺酸、酪胺酸、天冬醯胺、麵胺、天冬酸、麩胺酸、離胺酸、精胺酸、及、組胺酸。除非另行定義，否則如此處使用之「經保護」一詞包拉加成至氧、氮、硫或磷原子來防止其進一步反應或用於 ⑼其它目的之基團。寬廣多種氧及氮保護基為有機合成技藝界之熟諳技藝人士已知（參考Greene及Wuts，有機合成保護基，1999年，第3版，約輪威利父子公司，紐約）。除非另行陳明，芳基一詞如此處使用包括笨基、聯笨基或萘基且較佳為苯基。芳基視需要可經以〜個或多個部 29 200835507 分取代，該等取代部分諸如原子、經基、胺基、烧基胺基、芳基胺基、烧氧基、芳氧基、墙基、氰基、績酸、硫酸根、膦酸、磷酸根或膦酸根，或為未經保護，或如熟諳技藝人士已知可視需要經保護，例如如Greene等人，有機合成保 5 遵基’ 1991年’第2版’約翰威利父子公司，紐約教示。烷芳基或烷基芳基等詞包括有芳基取代基之烷基。芳烷基或芳基烷基等詞包括有烷基取代基之芳基。鹵原子一詞用於此處包括氯、漠、峨、及氟。醯基一詞包括羧酸酯，其中酯基之非羰基部分係選自 10於直鏈、分支鏈或環狀烷基或低碳烷基、烷氧基烷基包括甲氧基甲基、芳烧基包括苄基、芳氧基烧基諸如苯氧基甲基、芳基包括視需要可經以鹵素、（^至(:4燒基或(^至(:4烧氧基取代之苯基、磺酸酯諸如烷基磺醯基或芳烷基磺醯基包括甲磺醯基、一磷酸酯、二磷酸酯、或三磷酸酯、三苯 15甲基或一甲氧基三苯甲基、經取代之节基、三烷基矽烷基 (例如一甲基-弟二丁基石夕烧基）或二苯基甲基石夕烧基。醋之芳基最佳包含本基。「低碳醯基」一詞係指其中非幾基部分為低碳烷基之醯基。如此處使用，就對映異構純度而言，「實質上不含」或 20 「貫質上不存在有」一詞係指一種組成物包括至少85%或 90%重量比，較佳95%至98%重量比，又更佳99。/〇至100%重 1比所指定之對映異構物。於較佳實施例中，於本發明方法及化合物中，化合物實質上不含其它對映異構物。同理，刀離」一詞係指一種化合物組成物包括至少 30 200835507 85/〇或90/。重里比，較佳95%至98%重量比，及又更佳9州至100%重里比之化合物，差額包含其它化學種類或對映異構物。各自刀別」一詞用於此處係指分開施用之變數，該 5變數因應用用途而異有獨立變化。如此，於諸如R，，XYR” 之化口物中，其中R，，為「分別為碳或氮」，兩個r，，可為碳，兩個R”可為氮，或-個R，，為碳而另_個&，，為氮。藥學上可接受之鹽或前藥」一詞用於前文說明書中用來說明化合物之藥學上可接受之形式(諸如酷、鱗_、 ⑺鹽或相關基團其當投予病人時可提供該化合物。藥學上可接受之鹽包括衍生自藥學上可接受之無機驗或有機驗及無機酸或有機酸之鹽。適當鹽類包括衍生自驗金屬如钟及鈉之鹽及衍生自鹼土金屬如鈣及鎂之鹽，多種其它酸為热諳製藥業界人士眾所周知。藥學上可接受之前藥係指一種化合物其於宿主體内代謝，例如水解或氧化來形成本發明化合物。前藥之典型例包括於活性化合物之官能部分上具有生物不安定性保護基之化合物。前藥包括可經氧化、還原、胺化、去胺化、羥化、去羥化、水解、去水解、烷化、去烷化、醯化、去醯化、磷酸化、去磷酸化來製造活 2〇丨生化合物之化合物。「藥學上可接受之g旨類」一詞用於此處除非另行陳明’否則包括一種或多種化合物之酯類，該等酯類於審慎醫療判定之範疇内，適合用來與宿主組織接觸而無不當毒性、刺激性、過敏反應等，具有合理效益/風險比，可有效 31 200835507 用於其期望用途。「個體」一詞如此處使用係指動物，較佳為哺乳動物，最佳為人類。哺乳動物包括非人哺乳動物包括但非限於豬、羊、山羊、牛、鹿、騾、馬、猴及其它非人靈長類、 5 犬、貓、大鼠、小鼠、兔或任何其它已知或如此處揭示之哺乳動物。 5.2本發明化合物本發明提供TCN、TCN-P及相關化合物組合爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽用於治療增生 10 病症之特殊治療計劃之用途。如此處使用，除非另行指示，否則「曲西立濱化合物」及「曲西立濱及相關化合物」等詞係指有如下結構式之化合物：- A partial cancer metastasis. As used herein, unless otherwise indicated, the term alkyl includes, for example, 27 200835507 is a linear, branched or cyclic first hydrocarbon, second hydrocarbon, or third hydrocarbon of Cl to C24, particularly including methyl, Trifluoromethyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, pentyl, cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl , cyclohexyl, cyclohexylmethyl, 3-methylpentyl, 5 2,2-monomethylbutyl, and 2,3-dimethylbutyl. The alkyl group may be substituted, for example, with one or more substituents such as a halogen atom (F, C1, Br or I) (for example, CF3, 2-Br.ethyl, CH2F, CH2Q, CH2CF3, or CF2CF3). a hydroxyl group (e.g., CH2OH), an amine group (e.g., CH2NH2, CH2NHCH3, or CH2N(CH3)2), an alkylamino group, an arylamine group, an alkoxy 10 group, an aryloxy group, a stearyl group, an azide group (e.g., CH2N3), cyano (e.g., ch2CN), sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate, either unprotected, or as known to those skilled in the art, may be protected as desired, for example, as Greene et al. Organic Synthetic Protection Group, 1991, 2nd ed., John Wiley & Sons, New York's teachings, incorporated herein by reference. 15 Unless otherwise stated, the term lower alkyl as used herein means (^ to a saturated straight chain, a branched chain, or, if appropriate, a cyclic (eg cyclopropyl)alkyl group, including substituted forms. And unsubstituted forms. The term "alkylamino" or arylamino includes the radicals having one or two alkyl or aryl substituents. The term "mercaptoamino acid" includes both natural and synthetic ", yS". Amino acid, including but not limited to amino acids present in proteins, namely glycine, alanine, leucine, leucine, isoleucine, methionine, phenylalanine, tryptamine Acid, valine, serine, threonine, hemi-deaminic acid, _acid, aspartame, glutamine, aspartic acid, face acid, lysine, arginine, and 28 200835507 Amino acid. In a preferred embodiment, the amino acid is in the L-configuration. In addition, the amino acid < is a propyl group: a stilbene, an amidino group, an arginyl group, an isoleamine group, an anthracene Amidoxime, albino propylamine thiol, tryptamine sulfhydryl, egg amine sulfhydryl, glycidyl sulfhydryl, amide amine, sulphate, cysteamine, tyramine, aspartame 5 base Acrylamine, aspartame, pentanethiol, amidoxime, spermine thiol, histamine aryl, /3-propylamine thiol, chloroamine thiol, point-leucine thiol, 0- Isoamyl sulfhydryl, /3-aminoamine oxime, /9-phenylpropylamine oxime, tryptamine sulfhydryl, /5-egamine sulfhydryl, glycidinyl, phenanthine fluorenyl, stone - Sulfaguanidine,/5-cysteine thiol, tyramine sulfhydryl, aspartate, acetonyl 10 amine, 10,000-day sulfhydryl, /5-pentamethylene, /3- a derivative of an amidino group, a/5-spermine thiol group, or a point-histamine thiol group. When the term amino acid is used, it is regarded as a specific independent of a natural amino acid or a synthetic amino acid ester. Reveal that, but not limited to the D configuration, L configuration of glycine, alanine, lysine, leucine, isoleucine, methionine, phenylpropylamine Acid, 15 tryptophanic acid, valine acid, serine acid, threonine, cysteine, tyrosine, aspartame, face amine, aspartic acid, glutamic acid, lysine, spermine Acid, and, histidine. Unless otherwise defined, the term "protected" as used herein is added to oxygen, nitrogen, Or a phosphorus atom to prevent further reaction or for (9) other purposes. A wide variety of oxygen and nitrogen protecting groups are known to those skilled in the art of organic synthesis (see Greene and Wuts, Organic Synthetic Protective Groups, 1999, 3rd edition, about the round of Willie & Sons, New York). Unless otherwise stated, the term aryl is used herein to include stupid, phenyl or naphthyl and preferably phenyl. The aryl group can be used as needed. Substituted by one or more moieties 29 200835507, such substituted moieties such as atom, thiol, amine, alkylamino, arylamine, alkoxy, aryloxy, wall, cyano, acid , sulphate, phosphonic acid, phosphate or phosphonate, either unprotected or as known to those skilled in the art, may be protected as needed, for example, as Greene et al., Organic Synthetic 5 Compliance '1991' 2nd Edition 'John Wiley & Sons, New York teaches. The terms alkaryl or alkylaryl include alkyl groups having an aryl substituent. The terms arylalkyl or arylalkyl include aryl groups having an alkyl substituent. The term halogen atom is used herein to include chlorine, molybdenum, hydrazine, and fluorine. The term thiol includes carboxylic acid esters wherein the non-carbonyl moiety of the ester group is selected from the group consisting of 10 in a straight chain, a branched chain or a cyclic alkyl group or a lower alkyl group, and the alkoxyalkyl group includes a methoxymethyl group, a aryl group. The alkyl group includes a benzyl group, an aryloxyalkyl group such as a phenoxymethyl group, and an aryl group including a phenyl group which may be optionally substituted with a halogen, (^ to (: 4 alkyl group or (^ to (4) alkoxy group). A sulfonate such as an alkylsulfonyl or an aralkylsulfonyl group includes a methylsulfonyl group, a monophosphate, a diphosphate, or a triphosphate, a triphenyl 15 methyl group or a monomethoxytrityl group. a substituted sulfhydryl group, a trialkylalkylene group (for example, monomethyl-dibutyl fluorene) or a diphenylmethyl group; the aryl group of vinegar preferably contains a base. The term "thiol" refers to a fluorenyl group in which the non-base moiety is a lower alkyl group. As used herein, in terms of enantiomeric purity, "substantially free" or 20 "permanent does not exist" The term means a composition comprising at least 85% or 90% by weight, preferably 95% to 98% by weight, more preferably 99% to 100% by weight of the specified enantiomer. In a preferred embodiment, In the methods and compounds of the invention, the compound is substantially free of other enantiomers. Similarly, the term "knife separation" means a compound composition comprising at least 30 200835507 85 / 〇 or 90 /. by weight ratio, preferably 95 % to 98% by weight, and more preferably 9 to 100% by weight of the compound, the difference includes other chemical species or enantiomers. The term "individual" is used herein to mean a variable to be applied separately. The 5 variables vary independently depending on the application. Thus, in a rectification such as R, XYR, where R, is "carbon or nitrogen," respectively, two r, can be carbon, two The R" may be nitrogen, or - R, which is carbon and another _ &, is nitrogen. The term pharmaceutically acceptable salt or prodrug is used in the above description to describe the pharmaceutically active compound. An acceptable form (such as a cool, squamous, (7) salt or related group which provides the compound when administered to a patient. Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic and inorganic acids Or a salt of an organic acid. Suitable salts include derived metals such as clocks and sodium. And salts derived from alkaline earth metals such as calcium and magnesium, a variety of other acids are well known to those skilled in the pharmaceutical industry. A pharmaceutically acceptable prodrug is a compound which is metabolized, for example hydrolyzed or oxidized, to form a compound of the invention in a host. Typical examples of prodrugs include compounds having a biolabile protecting group on the functional moiety of the active compound. Prodrugs include oxidation, reduction, amination, deamination, hydroxylation, dehydroxylation, hydrolysis, dehydrolysis, Alkylation, dealkylation, deuteration, deuteration, phosphorylation, dephosphorylation to produce a compound of a living compound. The term "pharmaceutically acceptable g" is used herein unless otherwise 'Alternatively includes esters of one or more compounds that are suitable for use in contact with host tissues without undue toxicity, irritation, allergic reactions, etc., with reasonable benefit/risk ratio, within the scope of prudent medical judgment. Valid 31 200835507 for its intended use. The term "individual" as used herein refers to an animal, preferably a mammal, and most preferably a human. Mammals include non-human mammals including, but not limited to, pigs, sheep, goats, cows, deer, baboons, horses, monkeys, and other non-human primates, 5 dogs, cats, rats, mice, rabbits, or any other Know or a mammal as disclosed herein. 5.2 Compounds of the Invention The present invention provides the use of a combination of TCN, TCN-P and related compounds, such as gefitini, erlotinib or a salt thereof, for the treatment of a special treatment for a proliferative disorder. As used herein, unless otherwise indicated, the words "tricilide compound" and "tricilide and related compounds" refer to compounds having the following structural formula:

32 200835507 其中R2’、R3’及By分別為氫、視需要可經取代之磷酸根或膦酸根（包括一磷酸、二磷酸、或三磷酸或安定化磷酸前藥）；醯基（包括低碳醯基）；烷基（包括低碳烷基）；醯胺、磺酸酯包括烷基或芳基烷基；磺醯基包括甲磺醯基及苄 5基，其中该笨基視需要可經以一個或多個如於此處所述芳基之定義中說明之取代基取代；視需要可經取代之芳基磺醯基；脂質包括磷脂質；胺基酸；碳水化合物；胜肽；或膽固醇；或其它藥學上可接受之離去基，其於活體内玎提供一種化合物其中R2,、R，或r5,分別為Η或一磷酸、二磷 10 酸、或三磷酸；其中Rx及Ry分別為氫、視需要可經取代之磷酸根；醯基（包括低碳醯基）；醯胺、燒基（包括低碳炫基）；芳香族、去氧伸烷基諸如聚乙二醇、視需要可經取代之芳基磺醯基，脂質包括填脂質；胺基酸；碳水化合物；胜肽；或膽 15固醇；或其它藥學上可接受之離去基；於一個實施例中，該化合物係呈5’-鱗醚脂質或5’-鱗脂質投藥。32 200835507 wherein R2', R3' and By are respectively hydrogen, optionally substituted phosphate or phosphonate (including monophosphate, diphosphate, or triphosphate or stabilized phosphate prodrug); sulfhydryl (including low carbon) Alkyl (including lower alkyl); decylamine, sulfonate includes alkyl or arylalkyl; sulfonyl includes methanesulfonyl and benzyl 5, wherein the stupid group can be Substituted by one or more substituents as exemplified in the definition of aryl as described herein; optionally substituted arylsulfonyl; lipids including phospholipids; amino acids; carbohydrates; peptides; Cholesterol; or other pharmaceutically acceptable leaving group which provides a compound in which R 2 , R, or r 5 are respectively hydrazine or monophosphate, diphosphoric acid, or triphosphate; wherein Rx and Ry Respectively hydrogen, optionally substituted phosphate; sulfhydryl (including low carbon fluorenyl); decylamine, alkyl (including low carbon); aromatic, deoxyalkylene such as polyethylene glycol, An arylsulfonyl group which may be substituted as needed, the lipid includes a lipid-filling; an amino acid; carbon water a compound; a peptide; or a cholesterol steroid; or other pharmaceutically acceptable leaving group; in one embodiment, the compound is administered as a 5'-squaternary ether lipid or a 5'-squaric lipid.

Ri及R2各自分別為Η、視需要可經取代之直鏈、分支、或環狀烷基（包括低碳烷基）、：)：希基或炔基、CO-烷基、CO-烯基、CO-炔基、CO-芳基或雜芳基、c〇-烷氧基烷基、CO-20芳氧基烧基、CO-經取代之芳基、績醯基、炫基續醯基、芳基磺醯基、芳烧基績醯基。於一個實施例中，R2’及R3’為氫。於另一個實施例中， R2’及R5’為氫。於又另一個實施例中，r2,、r3,及r5,為氫。於又另一個實施例中，R2’、R3,、r5,、Ri&r2為氫。 33 200835507 於另一個實施例中，曲西立濱化合物具有如下結構式：Ri and R2 are each a linear, branched, or cyclic alkyl group (including a lower alkyl group) which may be substituted as needed, respectively:,: Hirschine or alkynyl, CO-alkyl, CO-alkenyl , CO-alkynyl, CO-aryl or heteroaryl, c〇-alkoxyalkyl, CO-20 aryloxyalkyl, CO-substituted aryl, fluorenyl, thiol , arylsulfonyl group, aromatic burning base. In one embodiment, R2' and R3' are hydrogen. In another embodiment, R2' and R5' are hydrogen. In yet another embodiment, r2, r3, and r5 are hydrogen. In yet another embodiment, R2', R3, r5, Ri&r2 are hydrogen. 33 200835507 In another embodiment, the triplicidin compound has the following structural formula:

其中R3為Η、視需要可經取代之直鏈、分支或環狀烷基 (包括低碳烷基）、烯基、或炔基、NH2、NHR4、N(R4)2、芳 5 基、烧氧基烧基、芳氧基烧基、或經取代之芳基；以及各個R4分別為Η、醯基包括低碳醯基、烧基包括低碳烧基諸如但非限於甲基、乙基、丙基、及環丙基、烯基、炔基、環烧基、烧氧基、烧氧基烧基、經基烧基、或芳基。於一次實施例中，R3為直鏈C1-11烷基、異丙基、第三丁基 10 或苯基。於一個實施例中，此處提供之曲西立濱化合物具有如下結構式：Wherein R 3 is a linear, branched or cyclic alkyl group (including a lower alkyl group), an alkenyl group, or an alkynyl group, NH 2 , NHR 4 , N(R 4 ) 2 , an aryl 5 group, and may be substituted as needed. An oxyalkyl group, an aryloxyalkyl group, or a substituted aryl group; and each of R 4 is an anthracene, a fluorenyl group including a lower fluorenyl group, and an alkyl group including a lower carbon group such as, but not limited to, a methyl group, an ethyl group, A propyl group, and a cyclopropyl group, an alkenyl group, an alkynyl group, a cycloalkyl group, an alkoxy group, an alkoxy group, a mercapto group, or an aryl group. In one embodiment, R3 is a linear C1-11 alkyl group, an isopropyl group, a tert-butyl group 10 or a phenyl group. In one embodiment, the triclinide compound provided herein has the formula:

34 200835507 於另一個實施例中，此處提供之曲西立濱化合物具有如下結構式：34 200835507 In another embodiment, the triclinide compound provided herein has the following structural formula:

f 於另一個實施例中，此處提供之曲西立濱化合物具有 5 如下結構式： %f In another embodiment, the triclinide compound provided herein has the following structural formula:

其中R6為Η、烷基（包括低碳烷基）、烯基、炔基、烷氧基烷基、羥基烷基、芳基烷基、環烷基、NH2、NHR4、NR4R4、 CF3、CH2OH、CH2F、CH2a、CH2CF3、C(Y3)3、C(Y3)2C(Y3)3、 10 C(=0)0H、C(=0)0R4、C(=0)-烷基、C(=0)·芳基、C(=0)· 烷氧基烷基、C(=0)NH2、C(=0)NHR4、C(=0)N(R4)2，此處各個Y3分別為H或鹵原子；以及 35 200835507 各個R4分別為Η、醯基包括低碳醯基、烷基包括低碳烷基諸如但非限於甲基、乙基、丙基、及環丙基、烯基、炔基、環烧基、烧氧基、烧氧基烧基、經基烧基、或芳基。於一次實施例中，R6為乙基、CH2CH2OH或CH2-苯基。於另一個實施例中，此處提供之曲西立濱化合物具有如下結構式：Wherein R6 is hydrazine, alkyl (including lower alkyl), alkenyl, alkynyl, alkoxyalkyl, hydroxyalkyl, arylalkyl, cycloalkyl, NH2, NHR4, NR4R4, CF3, CH2OH, CH2F, CH2a, CH2CF3, C(Y3)3, C(Y3)2C(Y3)3, 10 C(=0)0H, C(=0)0R4, C(=0)-alkyl, C(=0 · aryl, C(=0)·alkoxyalkyl, C(=0)NH2, C(=0)NHR4, C(=0)N(R4)2, where each Y3 is H or a halogen atom; and 35 200835507 each R4 is an anthracene, a fluorenyl group including a lower fluorenyl group, and an alkyl group including a lower alkyl group such as, but not limited to, a methyl group, an ethyl group, a propyl group, and a cyclopropyl group, an alkenyl group, an alkynyl group. , a cycloalkyl group, an alkoxy group, an alkoxy group, a base group, or an aryl group. In one embodiment, R6 is ethyl, CH2CH2OH or CH2-phenyl. In another embodiment, the triclinide compound provided herein has the following structural formula:

其中R7為Η、鹵原子、烷基（包括低碳烷基）、烯基、炔基、烧氧基、烧氧基院基、經基烧基、環烧基、墙基、氰 10 基、OH、OR4、NH2、NHR4、NR4R4、SH、SR4、CF3、CH2OH、 CH2F、CH2Q、CH2CF3、C(Y3)3、C(Y3)2C(Y3)3、C(=0)0H、 C(=0)0R4、C(=0)-烷基、C(=0)-芳基、C(=0)-烷氧基烷基、 C(=0)NH2、C(=0)NHR4、C(=0)N(R4)2*N3，此處各個 Y3 分別為H或鹵原子；以及 15 各個R4分別為Η、醯基包括低碳醯基、烷基包括低碳烷基諸如但非限於甲基、乙基、丙基、及環丙基、烯基、炔基、環烷基、烷氧基、烷氧基烷基、羥基烷基、或芳基。於一次實施例中，R7為甲基、乙基、苯基、氯或ΝΗ2。 36 200835507 於另一個實施例中，此處提供之曲西立濱化合物具有如下結構式：Wherein R7 is hydrazine, a halogen atom, an alkyl group (including a lower alkyl group), an alkenyl group, an alkynyl group, an alkoxy group, an alkoxy group, a carbyl group, a cycloalkyl group, a wall group, a cyanide group 10, OH, OR4, NH2, NHR4, NR4R4, SH, SR4, CF3, CH2OH, CH2F, CH2Q, CH2CF3, C(Y3)3, C(Y3)2C(Y3)3, C(=0)0H, C(= 0) 0R4, C(=0)-alkyl, C(=0)-aryl, C(=0)-alkoxyalkyl, C(=0)NH2, C(=0)NHR4, C( =0) N(R4)2*N3, where each Y3 is H or a halogen atom; and 15 each R4 is Η, 醯 includes a lower fluorenyl group, and the alkyl group includes a lower alkyl group such as but not limited to A Base, ethyl, propyl, and cyclopropyl, alkenyl, alkynyl, cycloalkyl, alkoxy, alkoxyalkyl, hydroxyalkyl, or aryl. In one embodiment, R7 is methyl, ethyl, phenyl, chloro or hydrazine. 36 200835507 In another embodiment, the triclinide compound provided herein has the following structural formula:

於另一個實施例中，此處提供之曲西立濱化合物具有 5 如下結構式：In another embodiment, the triclinide compound provided herein has the following structural formula:

如此處使用且除非另行指示，否則「表皮生長因子受體抑制劑」一詞係指靶定於表皮生長因子受體(EGFR)酪胺 37 200835507 酸激酶之化合物，該EGFR酪胺酸激酶於各種形式之癌症中高度表現且偶爾突變。於若干實施例中，化合物係以可逆方式結合至受體之腺苷三磷酸(ATP)結合位置。於若干實施例中，EGFR家族的兩個成員共同形成一個同質二元體。然 5後使用ATP分子來自我磷酸化彼此，造成其胞内結構的構型改變’暴露另一個結合位置來結合蛋白質，造成信號串級至細胞核。經由抑制ATP，自我磷酸化變成不可能，而信號中止°表皮生長因子受體抑制劑化合物之說明例包括爾洛堤尼系列化合物例如傑費堤尼及爾洛堤尼。〇如此處使用且除非另行指示，否則「爾洛堤尼系列化合物」一詞係指下式化合物：As used herein and unless otherwise indicated, the term "epidermal growth factor receptor inhibitor" refers to a compound that targets the epidermal growth factor receptor (EGFR) tyramine 37 200835507 acid kinase, which is a variety of compounds. Forms of cancer are highly expressed and occasionally mutated. In several embodiments, the compound binds reversibly to the adenosine triphosphate (ATP) binding site of the receptor. In several embodiments, two members of the EGFR family together form a homogeneous binary. After 5, the ATP molecules were used to phosphorylate each other, causing a change in the conformation of their intracellular structure to expose another binding site to bind the protein, causing the signal to cascade to the nucleus. By inhibiting ATP, autophosphorylation becomes impossible, and signaling halts. Illustrative examples of epidermal growth factor receptor inhibitor compounds include the erlotini series of compounds such as gefitini and erlotini. 〇 As used herein and unless otherwise indicated, the term "Llotini series of compounds" means a compound of the formula:

Ri及R2分別為氫、分別視需要可經取代之烷氧基、視 15需要可經取代之胺、芳香族胺、雜芳香族胺、視需要可經取代之直鏈、分支或環狀烷基； R3及R4各自分別為氫、分別為視需要可經取代之芳香族胺' 雜芳香族胺或環狀胺。於一具體實施例中，爾洛堤尼系列化合物包括但非限於下列結構式：Ri and R2 are each independently hydrogen, optionally substituted alkoxy, optionally substituted amine, aromatic amine, heteroaromatic amine, optionally substituted straight chain, branched or cyclic alkane Each of R3 and R4 is hydrogen, and an optionally substituted aromatic amine 'heteroaromatic amine or cyclic amine, respectively. In one embodiment, the erlotini series of compounds includes, but is not limited to, the following structural formula:

A傑費堤尼 B爾洛堤尼 38 200835507 須瞭解此處揭示之化合物可含有對掌中心。此種對掌中心可為(R)組態或（S)組態或可為其潙合物。如此，此處提供之化合物可為對映異構純質，或為立體異構混合物或非對映異構混合物。須瞭解此處揭示之化合物涵蓋任何外消 5旋形式、旋光形式、多形體形式或立體異構形式或其混合物，較佳具有此處所述之有用性質，技藝界眾所周知如何製備旋光形式，以及如何使用此處所述之標準試驗決定活性，或使用技藝界已知之其它類似試驗。可用於獲得化合物之光學異構物之方法實例包括下列： 10 (i)晶體之物理分離-可手勳分離個別對映異構物之巨觀晶體之技術。若存在有分開對映異構物之晶體，亦即材料為堆集物而各晶體於視覺上為分開，則可使用此項技術； (ii)同時結晶-唯有於材料為呈固態之堆集物時才可 15能之讓個別對映異構物分開由外消旋物溶體中分開結晶之技術； (iii) 酶催化光學分割_經由對映異構物與酶之不同反應速率達成外消旋物之部分分離或完全分離之技術； (iv) 酶催化非對稱性合成-呈少一個合成步驟係使用 20酶催化反應來獲得期望之對映異構物之對映異構純質映異構豐富合成前驅物之合成技術； 3 + ivj化学非對稱性合成_於町於產物中產生非對铲眭 (亦即對掌性)之條件下，由非對掌前驅物合成期望之冉構物之合成技術，該條件可使用對掌催化劑或對掌輔=異 39 200835507 達成； (vi)非對映異構物分離-外消旋化合物與對映異構純試劑(對f辅劑)反應，將個別對映異構物轉成非對映異構物之技術。所得非對映異構物隨後由於現在已經變成有較明顯的結構差異，故藉層析術或藉結晶分離，對掌辅劑隨後被去除來獲得期望之對映異構物；思 (VII； ‘、及非對稱性轉換-來自於外消旋与之非對映異構物平衡來於得自期望之對映異構物之非對日 10 15 20 異構物溶體中獲得優勢，或得自期望對映異構物之非對日異構物優先結晶擾亂平衡，最終原則上全部材料皆轉成斗自期望之對映異構物之結晶性非對映異構物之技術。期I, 之對映異構物隨後由非對映異構物中釋放； (vm)動態光學分割_本技術係指於動態條件下，藉由^ 、去構物與對掌非外消旋試劑或催化劑之不等反應速率光風心卜消旋物之部分群分職完全光學分割（或心尤予$割化合物之進一步光學分割）； :叫由非外消旋前驅物之對映異構特異性合成如處％物料獲得期望之對映異構物之—種合成技術，^ &化學之完好不因合成過程受損，或只有極少受損；盒靜L)#掌液相層析術-外消旋物之對映異構物藉由；相^91之差異交互作用而被分離人液體動相之技術1 發差異^材料所形成，或動相可含有額外對掌材料來^ 、父互作用； (χι)對掌氣相層析術-外消旋物氣化，對映異構物藉^ 40 200835507 其於氣體動相中與含有固定非外消旋對掌吸附相之管柱間之差異作用而分離；㈣使用對掌溶劑萃取_藉由—種對映異構物偏好溶解於-特定對掌溶劑來分離對映異構物之技術； 5 (Xm)跨對萃犋轉運·外消旋物放置與薄膜障壁接觸之技術。障壁典型分離兩種可相溶混之流體，-種含有外消旋物，驅動力諸如濃度差異或壓力差異造成跨該膜障壁之優先轉運。由於該膜之非外消旋對掌本質只允許外消旋物中之一種對映異構物通過的結果出現分離。 10 於若干實施例中，提供曲西立濱、磷酸曲西立濱 (TCN_P)、5’·磷酸曲西立濱（TCN_p)、或曲西立濱之〇娜加合物(TCN-DMF)。TCN可藉熟諳技藝人士已知之任一種技術合成，例如述於四面體函件，1971· 49:ρ· 4757_476〇。 TCN-P可藉熟諳技藝人士已知之任一項技術合成，例如說 15明於美國專利案4，123,524。TCN-DMF之合成例如說明於 INSERM，1978· 81··ρ·37-82。其它如此處所述之TCN相關化合物例如可根據揭示於下列之方法合成：GU(jmundsson K.S. 等人，核苷、核苷酸、核酸，2001. 20(10-11): p.1823-1830; Porcari A.R.等人，J Med Chem，2000. 43(12): ρ·2457-2463; 20 Porcari A.R·等人，核苷、核苷酸，1999. 18(11-12》 p.2475-2497; Porcari A.R·等人，J Med Chem，2000· 43(12): p.2438-2448; Porcari A.R.等人，核苷、核苷酸、核酸，2003, 22(12): p.2171-2193; Porcari A.R·等人，核苷、核苷酸、核酸，2004, 23(1-2): p.31-39; Schweinsberg P.D·等人，Biochem 41 200835507A gefitini B lodini 38 200835507 It is to be understood that the compounds disclosed herein may contain the center of the palm. Such a palm center can be either (R) configured or (S) configured or can be a compound thereof. Thus, the compounds provided herein may be enantiomerically pure or in stereoisomeric or diastereomeric mixtures. It is to be understood that the compounds disclosed herein encompass any of the free 5 s form, optically active form, polymorphic form or stereoisomeric form, or mixtures thereof, preferably having the useful properties described herein, and it is well known in the art how to prepare optically active forms, and How to use the standard assays described herein to determine activity, or to use other similar assays known to the art. Examples of the method which can be used to obtain the optical isomer of the compound include the following: 10 (i) Physical separation of crystals - A technique for separating macroscopic crystals of individual enantiomers. If there are crystals with separate enantiomers, ie the material is a heap and the crystals are visually separated, then this technique can be used; (ii) simultaneous crystallization - only if the material is a solid stack The technique of separating individual enantiomers from the racemic solution by crystallization; (iii) Enzymatic catalyzed optical cleavage _ via the different reaction rates of enantiomers and enzymes Partial separation or complete separation of the spins; (iv) Enzymatically catalyzed asymmetric synthesis - one less synthetic step using a 20 enzyme catalyzed reaction to obtain the enantiomerically pure representation of the desired enantiomer Synthetic technology for the synthesis of synthetic precursors; 3 + ivj chemical asymmetry synthesis _ in the production of non-pairing shovel (that is, the palm of the hand) in the product, the desired structure is synthesized by the non-pseudo-precursor Synthetic technology of the substance, which can be achieved by using the catalyst or the palm-assisted = iso 39 200835507; (vi) separation of diastereomers - racemic compounds and enantiomerically pure reagents (for f adjuvants) Reaction, converting individual enantiomers into diastereomers Surgery. The resulting diastereomers are then subsequently converted to a more pronounced structural difference, so by chromatography or by crystallization separation, the palm adjuvant is subsequently removed to obtain the desired enantiomer; ', and asymmetry conversion - from racemic and diastereomeric equilibrium to gain an advantage in the non-Japanese 10 15 20 isomer solution obtained from the desired enantiomer, or The non-paradox isomers from the desired enantiomers preferentially crystallize and balance the equilibrium, and in principle all materials are converted into a crystalline diastereomer from the desired enantiomer. I, the enantiomer is then released from the diastereomer; (vm) dynamic optical segmentation - the technique refers to the non-racemic reagent by ^, destructor and palm Or the unequal reaction rate of the catalyst, the partial group of the racemate, the racemate, the complete optical division (or the further optical division of the compound); the enantiospecific specificity of the non-racemic precursor Sex synthesis, such as the % material to obtain the desired enantiomer - a synthetic technique , ^ & chemical integrity is not damaged by the synthesis process, or only minimal damage; box static L) # palm liquid chromatography - the enantiomer of the racemate by; The technique of interaction is separated by the liquid phase of the human body. The difference is formed by the material, or the moving phase may contain additional pairs of palm materials to form a parental interaction; (χι) to palm gas chromatography-racemate Gasification, enantiomers are separated by the difference between the gas phase and the column containing the fixed non-racemic adsorption phase of the palm; (4) using the solvent extraction of the palm_ Enantiomers prefer a technique for the separation of enantiomers by the use of a specific solvent for the palm of the hand; 5 (Xm) cross-collection of the transport of the rhodium and the placement of the racemate with the barrier of the membrane barrier. The barrier typically separates two miscible fluids, containing racemates, and driving forces such as concentration differences or pressure differences cause preferential transport across the membrane barrier. Since the non-racemic nature of the membrane allows only one of the enantiomers in the racemate to pass through, the separation occurs. In several embodiments, triclinibine, trimethoprim phosphate (TCN_P), 5'. Tricineribin phosphate (TCN_p), or triclinide adduct (TCN-DMF) is provided. . TCN can be synthesized by any technique known to those skilled in the art, for example, in a tetrahedral letter, 1971 49: ρ· 4757_476〇. TCN-P can be synthesized by any of the techniques known to those skilled in the art, for example, in U.S. Patent No. 4,123,524. The synthesis of TCN-DMF is described, for example, in INSERM, 1978. 81·ρ·37-82. Other TCN-related compounds as described herein can be synthesized, for example, according to the methods disclosed below: GU (jmundsson KS et al., Nucleosides, Nucleotides, Nucleic Acids, 2001. 20(10-11): p. 1823-1830; Porcari AR et al, J Med Chem, 2000. 43(12): ρ·2457-2463; 20 Porcari AR et al., Nucleosides, Nucleotides, 1999. 18(11-12) p.2475-2497; Porcari AR et al, J Med Chem, 2000· 43(12): p. 2438-2448; Porcari AR et al, Nucleosides, Nucleotides, Nucleic Acids, 2003, 22(12): p.2171-2193; Porcari AR et al., Nucleosides, Nucleotides, Nucleic Acids, 2004, 23(1-2): p.31-39; Schweinsberg PD et al., Biochem 41 200835507

Pharmacol，1981· 30(18): p.2521-2526; Smith K.L·等人， Bioorg Med Chem Lett, 2004. 14(13): p.3517-3520;Pharmacol, 1981. 30(18): p.2521-2526; Smith K.L. et al., Bioorg Med Chem Lett, 2004. 14(13): p.3517-3520;

Townsend L.B.等人，核酸Symp Ser，1986· 1986 (17): P.41-44 ;及/或 Wotring LL·等人，癌症治療Rep，1986· 70(4): 5 p.491-7。 5.3藥學上可接受之鹽及前藥於化合物充分鹼性或酸性可形成安定之無毒酸鹽或鹼鹽之情況下呈藥學上可接受之鹽投予化合物為適當。藥學上可接受之鹽包括衍生自藥學上可接受之無機或有機鹼或 10 15 20 酸。適當鹽類包括衍生自鹼金屬諸如鉀及鈉、鹼土金屬諸乙酸鹽、檸檬酸鹽、丙二酸鹽、酒石酸鹽如鈣及鎂之鹽，多種其它酸為技藝界眾所周知。特別，藥學上可接受之鹽之實例為與可形成生理上可接受之陰離子之酸所形成之有機酸加成鹽，例如甲苯績酸鹽、甲續酸鹽、 k酸鹽、苯甲酸鹽、抗壞血酸鹽、α_酮基戊二酸鹽、及α•甘油基顧鹽。也可形成適當之無機酸鹽包括硫酸鹽、魏鹽酸鹽、及碳酸鹽。藥學上可接受之鹽也可使频藝界眾所周知之序獲，，例如充分驗性化合物諸如胺與適當之形成^理二二接文之陰離子之酸反應。也可製造竣酸之驗金屬（例如，，、鉀或鋰）鹽或鹼土金屬（例如鈣）鹽。此處所述之任___料可呈核㈣前藥活性、生物_率、蚊性心其它方式變更料之^同多觸酸前藥配體為已知。大致上㈣之一•、二貝鱗 42 200835507 酸或三磷酸之烷化、醯化或其它親脂改性將可提高核苷酸之安定性。可置換磷酸部分上之一個或多個氫之取代基之實例為烧基、芳基、類固醇、碳水化合物、包括糖類、i 2_ 二隨基甘油及醇類。多種取代基說明於R· Jones&N. 5 Bischofberger，抗病毒研究，1995. 27: ρ-1_Π。全部皆可與所揭示之核苷組合來達成期望之效果。於一個實施例中，曲西立濱或相關化合物提供作為5，_ 羥基親脂前藥。揭示適當親脂性取代基，其可共價摻混於核苷’較佳摻混於核苷製劑或親脂製劑之5’-〇h位置之美國 10 專利案之非限制性實例包括美國專利案5，149,794、 5，194,654、5,223,263、5,256,642、5,411，947、5,463,092、 5,543,389、5,543,390 ' 5,543,39卜及5,554,728，各案以引用方式併入此處。揭示可附接至本發明之曲西立濱或相關化合物之親脂 15 取代基或親脂製劑之外國專利申請案包括W0 89/02733、 W0 90/00555、W0 91/16920、W0 91/18914、W0 93/00910、 W0 94/26273、WO/15132、EP 0 350 287、EP 93917054.4、及 W0 91/1972 卜曲西立濱或相關化合物之衍生物之額外非限制性實例 2〇為含有如下公告案所述之取代基。此等經衍生之曲西立濱或相關化合物可用於内文所述之適應症，否則用作為抗病毒劑包括用作為抗-HIV劑或抗-HBV劑。Ho D.H.W.，Cancer Res” 1973 33: ρ·2816-2820; Holy Α·於抗病毒藥物設計進階，VoLI，De Clercq (編輯），JAI出版社，ρρ· 179-231; Hong 43 200835507 C.I·等人，Biochem Biophys Rs Commun，1979.88: p.l223-1229;Hong C.I.等人，J Med Chem，1980. 28: p.171-177; Hostetler Κ·Υ·等人，J Biol Chem，1990.266: p.11714-11717; Hostetler Κ·Υ·等人，Antiviral Res，1994.24: 5 p.59-67; Hostetler K.Y.等人，抗微生物劑化學治療，1994.38: P.2792-2797; Hunston R.N.等人，J Med Chem，1984.27: p.440-444; Ji Υ·Η·等人，J Med Chem，1990·33:ρ·2264·2270; Jones A.S_ 等人，J Chem Soc Perkin Trans，1984-1: p.1471-1474; Juodka B.A.及 Smart J·，Coll Czech Chem 10 Comm，1974·39:ρ·363-968; Kataoka S·等人，核酸研究Sym Ser, 1989.21: p.1-2; Kataoka S·等人，雜環，1991.32: p.1351-1356; KinchingtonD·等人，抗病毒Chem Chemother， 1992. 3: p.107-112; Kodama K.等人，日本癌症研究期刊， 1989， 80: p.679-685; Korty M.及 Engels J·， 15 Naunyn-Schmiedeberg’s Arch Pharmacol，1979.10: ρ·103-111; Kumar A.等人，J Med Chem，1990.33: ρ·2368·2375; LeBec C. 及 Huynh-dinh T.，四面體函件，1991.32: ρ·6553·6556; Lichtenstein J.等人，J Biol Chem，1960.235: ρ·457·465; Lucthy J·等人，Mitt Geg Lebensmittelunters Hyg，1981.72: 20 p.131-133 (Chem· Abstr· 95, 127093); McGuiganC·等人，核酸研究，1989.17: ρ·6065-6075; McGuigan C·等人，抗病毒 Chem Chemother，1990.1 :ρ· 107-113; McGuigan C•等人，抗病毒Chem Chemother，1990· 1: ρ·355·360; McGuigan C.等人，抗病毒Chem Chemother，1990· 1: ρ·25·33; McGuigan C. 44 200835507 等人，抗病毒研究，1991. 15: p.255-263; McGuigan C.等人，抗病毒研究，1992. 17: p.311-321; McGuigan C.等人，抗病毒Chem Chemother，1993. 4: ρ·97_101; McGuigan C·等人，J Med Chem，1993. 36: p.1048-1052。 5 抗-HIV劑AZT之烷基氫膦酸衍生物可比親代核苷類似物較為不具毒性。抗病毒Chem Chemother，5: 271-277; Meyer R.B.等人，四面體函件，1973· 269-272; NagyvaryJ· 等人，Biochem Biophys Res Commun，1973· 55: ρ·1072-1077; Namane A·等人，J Med Chem，1992· 35: ρ·3939_3044; 10 Nargeot J·等人，Natl. 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Interact， 1986. 57: p.347-355; Saneyoshi M.等人，Chem Pharm Bull， 1980· 28: p.2915-2923; Sastry J.K·等人，Mol Pharmacol， 5 1992· 41: p.441-445; Shaw J.P.等人，第 9屆 AAPS 年度會議， 1994年，加州聖地牙哥(摘要）。Shuto S.等人，四面體函件， 1987. 28: p.199-202; Shuto S·等人，ChemPharm Bull，1988· 36: ρ· 209-217。一種較佳磷酸前藥基團為S-醯基-2-硫乙基，也稱作為「SATE」。 10 可使用之前藥之額外實例係說明於下列專利案及專利申請案：美國專利案 5,614,548、5,512,671、5,770,584、 5,962,437、5,223,263、5,817,638、6,252,060、6,448,392、 5,411，947、5,744,592、5,484,809、5,827,831、5,696,277、 6,022,029、5,780,617、5，194,654、5,463,092、5,744,461、 15 4,444,766、4,562，179、4,599,205、4,493,832、4,221，732、 5，116,992、6,429,227、5，149,794、5,703,063、5,888,990、 4,810,697、5,512,67卜 6,030,960、2004/0259845、6,670,34卜 2004/0161398、2002/082242、5,512,671、2002/0082242及/ 或 PCT 公告案 WO 90/11079、WO 96/39197、及/或 WO 20 93/08807。 5.4活艘内功效/投藥計劃於本發明之另一個態樣中，提供用藥計劃其可限制 TCN及相關化合物之毒性副作用。於另一個實施例中，此種給藥計劃可減少或消除毒性副作用，包括但非限於肝毒 46 200835507 性、血小板減少、高血糖、嘔吐、低血鈣、貧血、低白蛋白血症、骨髓抑制、血中三酸甘油酯過高、血中澱粉梅過高、腹瀉、胃炎及/或發燒。於另一個實施例中，投予TCN、TCN-Ρ或相關化合物 5及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽提供於活體内於至少15%-20°/。個體之至少部分反應或完全反應。於特定實施例中，部分反應可為腫瘤之至少15、2〇、 25、30、35、40、50、55、60、65、70、75、80或 85%退行。於其它實施例中，本反應於接受治療處理病人之至少 10 15、15、20、25、30、35、40、50、55、60、65、70、75、 80、85或90%為顯著。於其它實施例中，可藉此處揭示之任一種治療計劃獲得此種反應率。於其它實施例中，提供已經被診斷患有癌症之個體，係經由根據一種投藥計劃將有效量之TCN、TCN-Ρ或相關 15化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽投予該個體，該投藥計劃包括投予曲西立濱化合物及/ 或爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽每週一次計三週，接著為一週之未投藥時間（亦即透過28天週期）。於其它實施例中，此種28天週期可重複至少2、3、4、 20 或5次，或重複至腫瘤之退行明顯為止。於其它實施例中，提供42天週期，其中此處揭示之化合物可每週一次投予計四週，接著為未投予曲西立濱化合物及/或爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽之兩週時間。於其它實施例中，此種42天週期可重複至 47 200835507 少2、3、4、或5次，或重複至腫瘤之退行明顯為止。於特定實施例中，低於12，低於11或低於10毫克/平方米TCN、 TCN-P或相關化合物可根據42天週期投予。於其它特定實施例中，2、3、4、5、6、7、8、9、10、或15毫克/平方米 5 TCN、TCN-P或相關化合物可根據42天週期投予。於另一個特定實施例中，投予約1毫克至約50毫克爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽。於特定實施例中，可根據42天週期投予1、5、10、15、20、25、30、35、或 40毫克爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其 10 鹽。於另一個實施例中，提供於個體治療癌症之方法，係經由對該個體投予一種給藥計劃，每週一次投予10毫克/平方米或以下之TCN、TCN-P或相關化合物及低於約30毫克爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽。於 15 特定實施例中，每週一次投予0.5、1、1.5、2、2.5、3、3.5、 4、4.5、5、5.5、6、6.5、7、7.5、8、8.5、9、9.5或 10毫克 /平方米如此處揭示之TCN、TCN-P或相關化合物。於另一個特定實施例中，可每週一次投予1、5、10、15、20、25、 30、35、或40毫克爾洛堤尼系列化合物例如傑費堤尼、爾 20 洛堤尼或其鹽。於本發明之實施例中，此處揭示之化合物可於短時間例如約5、10、15、20、30或60分鐘時間同時呈單一大劑量投予。於其它實施例中，提供投藥計劃，其中該等化合物係透過連續輸注歷至少24、48、72、96或120小時時間同時 48 200835507 投予。於若干實施例中，透過連續注射或大劑量注射投予曲西立濱化合物及/或爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可於某個頻率重複，該投藥頻率至少：每週一次、每兩週一次、每三週一次、每個月一次、每五 5 週一次、每六週一次、每八週一次、每十週一次、及/或每十二週一次。投藥類型及頻率可以此處所述之任一種方式組合來形成投藥週期。曲西立濱化合物及/或爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可透過某種投藥週期重複投予，例如呈大劑量注射每兩週一次歷時三個月。 10 投藥計劃可投予至少歷時：1、2、3、4、5、6、7、8、9、 10、U、12、18、或24個月。另外，可投予一病人至少2、 3、4、5、6、7、8、9、10、11、12、15或20個投藥週期。曲西立濱化合物及/或爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可根據此處揭示之任一種組合投予， 15 例如曲西立濱化合物及/或爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可每週一次，每三週共三個週期。於其它實施例中，化合物可每日至少一次投予歷時至少2、3、4、5、6、7、8、9、或10曰。此種投藥接著為未投予曲西立濱化合物及/或爾洛堤尼系列化合物例如傑費 20 堤尼、爾洛堤尼或其鹽之相對應週期時間。如此處揭示之TCN、TCN-P及相關化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可以可有效造成腫瘤退行之數量投予病人。TCN、TCN-P及相關化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽之 49 200835507 投予可提供於活體内於至少15-20%個體之至少部分反應，諸如至少15%、20%或30%反應或完全反應。於若干實施例中，至少2、5、10、15、20、30或50毫克/平方米此處揭示之曲西立濱化合物可投予一個體。於若干實施例中，至少 5 約0.5、1、1.5、2、2.5、3、3.5、4、4.5、5、5.5、6、6.5、 7、7.5、8、8.5、9、9.5、10、12、15、17、20、25、30、 35、40、45、50、55、60、65、70、75、80、85、90、95、 1〇〇、150、165、175、200、250、300、或350毫克/平方米如此處揭示之TCN、TCN-P或相關化合物可投予一個體。 10 於右干實施例中，1、5、10、15、20、25、30、35或40毫克爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可投予一個體。化合物之投予可根據此處揭示之任一種治療計劃進行。於特定實施例中，投藥計劃包括同時、循序或於一段 15 時間投予低於約20毫克/平方米TCN及相關化合物及少於約 30毫克爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽。於一個實施例中，低於20毫克/平方米TCN或相關化合物可與少於約3〇毫克爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽同時每週一次投予。於另一個實施例中’ 20低於20毫克/平方米TCN或相關化合物可每週一次投予’而少於約30毫克爾洛堤尼系列化合物例如傑費堤尼、爾洛权尼或其鹽可於下週投予。於額外實施例中，2毫克/平方米、5毫克/平方米、lG 毫克/平方米及/或15毫克/平方米TCN或相關化合物及少於 50 200835507 約300、250、200、150或loo毫克爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可投予一個體。於另一個實施例中，y於克/平方米曲西立濱化合物及少於約如〇毫克爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可 5透過連續輸注歷至少5日投予一個體。本發明提供如此處揭示之投藥類型、頻率、週期數及劑量之任一種組合。 5.5病人族群之篩檢於本發明之另-個態樣中，提供方法來識別對曲西立濱(TCN)及相關化合物之毒性效應敏感之癌症或腫瘤。於一 10個貫鈀例中，提供於哺乳動物治療癌症或腫瘤之方法，係經由（i)由該腫瘤獲得生物樣本；⑼判定該癌症或腫瘤是否過度表現Akt激酶或高度活化之且磷酸化之Akt激酶，以及㈣以如此處所述之曲西立濱或相關化合物治療該癌症或腫瘤。於-個實施例中，生物樣本可為活體組織檢查。於 15其它實施例中，生物樣本可為得自該腫瘤或癌症之流體、細胞及/或抽吸物。生物樣本可根據Μ技藝人士已知之任-種技術獲得。於-個實施例中，可進行活體組織檢查來獲得生物樣本。活體組織檢查為由體内移出組織或細胞供檢查所進行之程序。此種活體組織檢查可於診療室内進行但有其它可能需要於醫院設備中進行。此外，某些活體組織檢查可能要求麻醉來造成該區域喪失知覺，而其它檢查無需任何麻醉。於若干實施例中，可進行内視鏡活體組織檢查。此類型之活體組織檢查係透過纖維鏡檢内視鏡（一根細長薄 51 200835507 管於末财密切對焦望遠鏡供觀看）穿過自然身體孔口（亦即直腸）或小的切口（亦即關節鏡檢）來進行。内視鏡用來觀看有問題的器官是否有異常區或可疑區，俾便獲得小量組織供研究。内視鏡手術係依據欲觀看及/或欲治療之器官或 5身體區域來命名。醫師將内視鏡***胃腸道（消化道鏡檢）、膀胱(膀胱鏡檢）、腹腔(腹腔鏡檢）、關節腔（關節鏡檢）、胸腔中部（縱膈鏡檢）、或氣管與支氣管系統(咽頭鏡檢及支氣管鏡檢）。於另一個實施例中，施行骨髓活體組織檢查。此類型 1〇活體組織檢查可由胸骨或體脊體骨（後背骨纟兩側的骨區）施行。清潔皮膚，給予局部麻醉劑來麻痺該區。細長剛硬針***骨髓内，抽吸細胞進行研究；此步驟偶爾導致不適。中心活體組織檢查（由骨髓取出小的骨「碎片」）可遵照抽吸程序。 15 於另一實施例中，切除或切開活體組織檢查可於哺乳動物施行。當需要較寬或較深的皮膚部分時常使用此型活體組織檢查。使用手術刀，取下完整皮膚厚度供進一步檢查，及縫合傷口（使用手術縫線來縫合關閉）。當整個腫瘤被移除時，稱作為切除活體組織檢查技術。當只移除部分腫 2〇瘤時’稱作為切開活體組織檢查技術。例如當懷疑為黑素瘤（一類皮膚癌）時經常以切除活體組織檢查為佳。又有其它實施例巾’可使肖細針純(F N A)活體組織檢查。此型活體組織檢查使用-根細針來由腫瘤移出極小塊組織。偶爾使用局部麻醉來麻痺該區，但本程序罕見造成 52 200835507 重大不適且不留疱。例如FNA並未用於可疑的癌的檢查，但可用於檢查黑素瘤附近的大型淋巴結，來瞭解黑素瘤是否已經轉移(擴散）。電腦斷層掃描(CT掃描或CAT掃描）可用來將細針導引入内臟器官諸如肺臟或肝臟的腫瘤。 5 於其它實施例中，可進行衝孔、剃除及/或皮膚活體組織檢查。衝孔活體組織檢查涉及使用生檢儀器取出一段短圓柱體或稱作「蘋果心」組織來取較深皮膚樣本。投予局部麻醉後，儀器於皮膚表面上旋轉至儀器穿過各層，包括真皮、表皮、和皮下的最表淺部分（皮下脂肪）。剃除活體組 10織檢查涉及藉刮除來移出皮膚的頂層。剃除活體組織檢查也係採用局部麻醉進行。皮膚活體組織檢查涉及取出皮膚樣本於顯微鏡下檢查來判定是否存在有黑素瘤。活體組織檢查係於局部麻醉下進行。於特定實施例中，提供判定腫瘤是否過度表現Akt激酶 15 之方法。Akt激酶之過度表現可參照激酶之磷酸化狀態。Akt 之高度鱗酸化可根據此處所述方法檢測。於一個實施例中，腫瘤活體組織檢查可與對照組織作比較。對照組織可為來自於進行活體組織檢查之哺乳動物的正常組織、或得自健康哺乳動物之正常組織。Akt激酶過度表現或高度鱗酸 20 化可判定腫瘤活體組織檢查含有比較對照組織更大量的 Akt激酶及/或Akt激酶鱗酸化，諸如比對照組織所含之Akt 激酶至少約 1.5、2、2.25、2.5、2.75、3、3.25、3.5、3.75、 4、4.25、4.5、4.75 ' 5、5.5、6、7、8、9、或1〇倍大量之Townsend L. B. et al., Nucleic Acid Symp Ser, 1986 1986 (17): P. 41-44; and/or Wotring LL et al., Cancer Therapy Rep, 1986 70(4): 5 p.491-7. 5.3 Pharmaceutically Acceptable Salts and Prodrugs It is appropriate to administer the compound as a pharmaceutically acceptable salt in the case where the compound is sufficiently basic or acidic to form a stable non-toxic acid or base salt. Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic bases or 10 15 20 acids. Suitable salts include those derived from alkali metals such as potassium and sodium, alkaline earth metal acetates, citrates, malonates, tartrates such as calcium and magnesium, and a variety of other acids are well known in the art. In particular, examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids which form physiologically acceptable anions, such as toluic acid salts, methylation salts, k acid salts, benzoates. , ascorbate, α-ketoglutarate, and α•glyceryl salt. Suitable inorganic acid salts can also be formed, including sulfates, sulphates, and carbonates. The pharmaceutically acceptable salts are also well known in the art, for example, by fully reacting a compound such as an amine with an acid which forms an anion of the appropriate composition. It is also possible to produce a metal (e.g., potassium, or lithium) or alkaline earth metal (e.g., calcium) salt of tannic acid. Any of the ___ materials described herein may be nuclear (IV) prodrug activity, bio-rate, mosquito heart, other means of change. The poly-touch acid prodrug ligand is known. Substantially (4) One, two shell scales 42 200835507 Alkylation, deuteration or other lipophilic modification of acid or triphosphate will increase the stability of the nucleotide. Examples of substituents which may replace one or more hydrogens on the phosphoric acid moiety are an alkyl group, an aryl group, a steroid, a carbohydrate, including a saccharide, i 2 -dihexyl glycerin, and an alcohol. A variety of substituents are described in R. Jones & N. 5 Bischofberger, Antiviral Research, 1995. 27: ρ-1_Π. All can be combined with the disclosed nucleosides to achieve the desired effect. In one embodiment, triclinide or a related compound is provided as a 5,-hydroxy lipophilic prodrug. A suitable lipophilic substituent is disclosed which can be covalently incorporated into a nucleoside. A non-limiting example of a US patent that is preferably incorporated in the 5'-〇h position of a nucleoside formulation or a lipophilic formulation includes a US patent case. 5, 149, 794, 5, 194, 654, 5, 223, 263, 5, 256, 642, 5, 411, 947, 5, 463, 092, 5, 543, 389, 5, 543, 390 '5, 543, 39, and 5, 554, 728, each of which is incorporated herein by reference. Disclosed are lipophilic 15 substituents or lipophilic formulations that can be attached to the tromethine or related compounds of the invention, including W0 89/02733, W0 90/00555, W0 91/16920, W0 91/18914 Further non-limiting examples of WZ 93/00910, W0 94/26273, WO/15132, EP 0 350 287, EP 93917054.4, and W0 91/1972 derivatives of flurazepam or related compounds are as follows Substitutes as stated in the announcement. Such derived triclinide or related compounds can be used in the indications described herein, or as anti-viral agents, including as anti-HIV agents or anti-HBV agents. Ho DHW, Cancer Res” 1973 33: ρ·2816-2820; Holy 进·Advanced in the design of antiviral drugs, VoLI, De Clercq (editor), JAI Press, ρρ· 179-231; Hong 43 200835507 CI·etc. Human, Biochem Biophys Rs Commun, 1979.88: p.l223-1229; Hong CI et al, J Med Chem, 1980. 28: p. 171-177; Hostetler Κ·Υ· et al, J Biol Chem, 1990.266: p. 11714-11717; Hostetler Κ·Υ· et al, Antiviral Res, 1994.24: 5 p.59-67; Hostetler KY et al., Antimicrobial Chemotherapy, 1994.38: P. 2792-2797; Hunston RN et al., J Med Chem, 1984.27: p.440-444; Ji Υ·Η· et al, J Med Chem, 1990·33: ρ·2264·2270; Jones A.S_ et al., J Chem Soc Perkin Trans, 1984-1: p .1471-1474; Juodka BA and Smart J·, Coll Czech Chem 10 Comm, 1974·39: ρ·363-968; Kataoka S· et al., Nucleic Acids Research Sym Ser, 1989.21: p.1-2; Kataoka S· Et al., Heterocycle, 1991.32: p. 1351-1356; Kinchington D. et al., Antiviral Chem Chemother, 1992. 3: p. 107-112; Kodama K. et al., J. Cancer Research Journal, 1989, 80: p .679-685; Korty M. And Engels J., 15 Naunyn-Schmiedeberg's Arch Pharmacol, 1979.10: ρ·103-111; Kumar A. et al., J Med Chem, 1990.33: ρ·2368·2375; LeBec C. and Huynh-dinh T., tetrahedron Letter, 1991.32: ρ·6553·6556; Lichtenstein J. et al., J Biol Chem, 1960.235: ρ·457·465; Lucthy J. et al., Mitt Geg Lebensmittelunters Hyg, 1981.72: 20 p. 131-133 (Chem· Abstr. 95, 127093); McGuigan C. et al., Nucleic Acids Research, 1989.17: ρ·6065-6075; McGuigan C· et al., Antiviral Chem Chemother,1990.1: ρ·107-113; McGuigan C• et al., Antiviral Chem Chemother, 1990· 1: ρ·355·360; McGuigan C. et al., Antiviral Chem Chemother, 1990· 1: ρ·25·33; McGuigan C. 44 200835507 et al., Antiviral Research, 1991. 15: P.255-263; McGuigan C. et al., Antiviral Research, 1992. 17: p.311-321; McGuigan C. et al., Antiviral Chem Chemother, 1993. 4: ρ·97_101; McGuigan C· et al. J Med Chem, 1993. 36: p. 1048-1052. 5 The alkyl-hydrogen phosphonic acid derivative of the anti-HIV agent AZT is less toxic than the parent nucleoside analog. Antiviral Chem Chemother, 5: 271-277; Meyer RB et al., Tetrahedron Letter, 1973·269-272; Nagyvary J. et al., Biochem Biophys Res Commun, 1973· 55: ρ·1072-1077; Namane A·etc. Man, J Med Chem, 1992· 35: ρ·3939_3044; 10 Nargeot J. et al., Natl. Acad. Sci. USA, 1983.80: p. 2395-2399; Nelson Κ·Α· et al., J Am Chem Soc , 1987.109: p. 4058-4064; Nerbonne J. M. et al., Nature, 1984· 301: p. 74-76; Neumann JM et al., J Am Chem Soc, 1 1989. Ill: p. 4270-4277; Ohno R. et al., Oncology, 1991. 48: ρ·451-455; 15 Palomino E. et al., J Med Chem, 1989· 32: p. 622-625; Perkins RM· et al., Antiviral Research, 1993 20 (Suppl. I): ρ·84; Piantadosi C. et al., J Med Chem, 1991. 34: 1408-1414; Pompon A. et al., Antiviral Chem Chemother, 1994· 5: ρ·91·98 PostemarkT·, Anu Rev Pharmacol, 1974· 14: ρ·23-33; Prisbe EJ· et al, J 20 Med Chem, 1986· 29: p.671-675; Pucch F. et al., Antiviral Research, 1993 22: p.155-174; Pugaeva VR et al., Gig Trf Prof Zabol, 1969, 13: ρ 47·48 (Chem. Abstr. 72, 212); Robins RK·, Pharm Res, 1984·11-18; Rosowsky A. et al., J Med Chem, 1982. 25: p.l71,178; Ross W·, Biochem Pharm, 1961· 45 200835507 8: p. 235-240; Ryu EK et al, J Med Chem, 1982. 25: p. 1322-1329; Saffhill R. and Hume WJ·, Chem Biol. Interact, 1986. 57 : p.347-355; Saneyoshi M. et al., Chem Pharm Bull, 1980·28: p.2915-2923; Sastry JK et al., Mol Pharmacol, 5 1992· 41: p.441-445; Shaw JP et al. People, 9th AAPS Annual Meeting, 1994, San Diego, California (Abstract). Shuto S. et al., Tetrahedron Letter, 1987. 28: p. 199-202; Shuto S. et al., Chem Pharm Bull, 1988. 36: ρ· 209-217. A preferred phosphoric acid prodrug group is S-mercapto-2-thioethyl, also known as "SATE". 10 Additional examples of prodrugs that can be used are described in the following patents and patent applications: U.S. Patent Nos. 5,614,548, 5,512,671, 5,770,584, 5,962,437, 5,223,263, 5,817,638, 6,252,060, 6,448,392, 5,411,947, 5,744,592, 5,484,809, 5,827,831, 5,696,277, 6,022,029,5,780,617,5,194,654, 5,463,092, 5,744,461, 15 4,444,766, 4,562,179, 4,599,205, 4,493,832, 4,221,732, 5,116,992, 6,429,227, 5,149,794, 5,703,063, 5,888,990, 4,810,697, 5,512,67, 6,030,960, 2004 </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> <RTIgt; 5.4 Live Intra-Effective/Dosing Plan In another aspect of the invention, a medication regimen is provided which limits the toxic side effects of TCN and related compounds. In another embodiment, such a dosing schedule can reduce or eliminate toxic side effects including, but not limited to, hepatotoxicity 46 200835507 Sex, thrombocytopenia, hyperglycemia, vomiting, hypocalcemia, anemia, hypoalbuminemia, bone marrow Inhibition, high triglycerides in the blood, high starch plum in the blood, diarrhea, gastritis and/or fever. In another embodiment, the administration of TCN, TCN-oxime or related compound 5 and the erlotini series of compounds such as gefitini, erlotini or a salt thereof is provided in vivo at least 15%-20°/ . At least partial or complete response of an individual. In a particular embodiment, the partial response can be at least 15, 2, 25, 30, 35, 40, 50, 55, 60, 65, 70, 75, 80 or 85% regression of the tumor. In other embodiments, the response is at least 10 15, 15, 20, 25, 30, 35, 40, 50, 55, 60, 65, 70, 75, 80, 85 or 90% of the patient being treated for treatment is significant . In other embodiments, such a response rate can be obtained by any of the treatment plans disclosed herein. In other embodiments, providing an individual who has been diagnosed with cancer is via an effective dosage of TCN, TCN-Ρ or related 15 compounds and a compound of the Llotini series, such as Jerepini, Erlo, according to a dosing schedule Dini or its salt is administered to the individual, and the dosing schedule includes administration of the triclinide compound and/or the erlotini series of compounds such as gefitini, erlotini or its salt for three weeks a week. This is followed by a week of unmedicated time (ie, through a 28-day cycle). In other embodiments, such a 28 day cycle can be repeated at least 2, 3, 4, 20 or 5 times, or repeated until the tumor is regressed significantly. In other embodiments, a 42-day cycle is provided wherein the compound disclosed herein can be administered once a week for four weeks, followed by failure to administer the triclinib compound and/or the erlotini series of compounds such as gefitini Two weeks of lodoti or its salt. In other embodiments, such a 42-day cycle can be repeated to 47, 2008, 35,507 less than 2, 3, 4, or 5 times, or until the tumor retreat is apparent. In a particular embodiment, less than 12, less than 11 or less than 10 mg/m 2 of TCN, TCN-P or related compounds can be administered according to a 42 day period. In other specific embodiments, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 15 mg/m 2 5 TCN, TCN-P or related compounds can be administered according to a 42 day period. In another specific embodiment, from about 1 mg to about 50 mg of erlotinib series of compounds such as gefitini, erlotini or a salt thereof are administered. In a particular embodiment, 1, 5, 10, 15, 20, 25, 30, 35, or 40 mg of erlotini series compounds such as gefitini, erlotini or its 10 salt. In another embodiment, the method for treating cancer in an individual is by administering to the individual a dosing schedule, once a week, administering TCN, TCN-P or related compounds at 10 mg/m or less and low. About 30 mg of erlotini series compounds such as gefitini, erlotini or a salt thereof. In a particular embodiment, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 mg/m 2 TCN, TCN-P or related compounds as disclosed herein. In another specific embodiment, 1, 5, 10, 15, 20, 25, 30, 35, or 40 mg of erlotini series compounds such as Gefitini and Er 20 Lotini can be administered once a week. Or its salt. In an embodiment of the invention, the compounds disclosed herein can be administered in a single large dose simultaneously for a short period of time, e.g., about 5, 10, 15, 20, 30 or 60 minutes. In other embodiments, a dosing schedule is provided wherein the compounds are administered by continuous infusion for at least 24, 48, 72, 96 or 120 hours while 48 200835507. In several embodiments, the administration of the triclinide compound and/or the erlotinib series compound, such as gefitinib, erlotini or a salt thereof, by continuous injection or bolus injection can be repeated at a certain frequency, Dosing frequency at least: once a week, once every two weeks, once every three weeks, once a month, every five five weeks, every six weeks, every eight weeks, every ten weeks, and / or every twelve Once a week. The type and frequency of administration can be combined in any of the ways described herein to form a dosing cycle. The triclinide compound and/or the erlotini series of compounds such as gefitinib, erlotinib or a salt thereof can be repeatedly administered through a certain administration cycle, for example, in a large dose every two weeks for three months. . 10 The dosing plan can be administered for at least one of the following: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, U, 12, 18, or 24 months. Alternatively, a patient may be administered at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15 or 20 dosing cycles. The triclinide compound and/or the erlotini series of compounds such as gefitinib, erlotinib or a salt thereof can be administered according to any combination disclosed herein, 15 such as triclinide compound and/or er The Lotini series of compounds such as Gefitini, Llotini or their salts can be used once a week for three cycles every three weeks. In other embodiments, the compound can be administered at least once a day for at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 曰. Such administration is followed by a corresponding cycle time for the non-administration of the triclinide compound and/or the erlotini series of compounds such as Jeffrey 20, Leroti or its salt. TCN, TCN-P and related compounds as disclosed herein and the erlotini series of compounds such as gefitinib, erlotinib or a salt thereof can be administered to a patient in an amount effective to cause tumor regression. TCN, TCN-P and related compounds and the erlotini series of compounds such as gefitini, erlotini or its salts 49 200835507 can be administered at least in part to at least 15-20% of the individual in vivo, Such as at least 15%, 20% or 30% reaction or complete reaction. In some embodiments, at least 2, 5, 10, 15, 20, 30 or 50 mg/m 2 of the triclinide compound disclosed herein can be administered to a single body. In some embodiments, at least 5 are about 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 12 , 15, 17, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 1〇〇, 150, 165, 175, 200, 250 300, or 350 mg/m 2 TCN, TCN-P or related compounds as disclosed herein may be administered to one body. 10 In the right-hand embodiment, 1, 5, 10, 15, 20, 25, 30, 35 or 40 mg of the erlotini series of compounds such as gefitini, erlotini or a salt thereof can be administered to one body . Administration of the compound can be carried out according to any of the treatment plans disclosed herein. In a particular embodiment, the dosing schedule comprises administering less than about 20 mg/m2 of TCN and related compounds and less than about 30 mg of erlotini series of compounds, such as Gefitini, at the same time, sequentially or over a period of 15 hours. Lotini or its salt. In one embodiment, less than 20 mg/m 2 of TCN or related compound may be administered once a week with less than about 3 mg of lovastini series of compounds such as gefitini, lodoti or its salt. . In another embodiment, '20 less than 20 mg/m 2 of TCN or related compound can be administered once a week' and less than about 30 mg of erlotini series of compounds such as gefitini, erlocini or its Salt can be administered next week. In additional embodiments, 2 mg/m2, 5 mg/m2, lG mg/m2 and/or 15 mg/m2 TCN or related compounds and less than 50 200835507 about 300, 250, 200, 150 or loo A milligram of the lodotinic series of compounds such as gefitini, erlotini or a salt thereof can be administered to one body. In another embodiment, y gram per square meter of triclinide compound and less than about 〇〇尔尔堤系列系列系列系列系列例如杰杰杰杰或其或其或其或其或其或其或其或其或其或其或其或其或其或其或其或其或其或其或其或其Give one body for at least 5 days. The present invention provides any combination of the type of administration, frequency, number of cycles, and dosage as disclosed herein. 5.5 Screening of Patient Populations In another aspect of the invention, methods are provided to identify cancers or tumors that are sensitive to the toxic effects of Tricinem (TCN) and related compounds. In a method of treating a cancer or a tumor in a mammal, by (i) obtaining a biological sample from the tumor; (9) determining whether the cancer or tumor overexpresses Akt kinase or highly activated and phosphorylated. Akt kinase, and (d) treating the cancer or tumor with triplicin or a related compound as described herein. In one embodiment, the biological sample can be a biopsy. In other embodiments, the biological sample can be a fluid, cell, and/or aspirate derived from the tumor or cancer. Biological samples can be obtained according to any technique known to those skilled in the art. In one embodiment, a biopsy can be performed to obtain a biological sample. Biopsy is the procedure performed by the removal of tissue or cells from the body for examination. Such biopsy can be performed in the treatment room but others may need to be performed in hospital equipment. In addition, some biopsy may require anesthesia to cause loss of consciousness in the area, while other tests do not require any anesthesia. In several embodiments, an endoscopic biopsy can be performed. This type of biopsy is passed through a fiberoptic endoscope (a thin thin 51G3535 in the end of the close focus telescope for viewing) through a natural body orifice (also known as the rectum) or a small incision (ie joint Microscopic examination). Endoscopes are used to see if the problematic organ has an abnormal or suspicious area, and a small amount of tissue is available for research. Endoscopic surgery is named according to the organ or body area to be treated and/or to be treated. The physician inserts the endoscope into the gastrointestinal tract (digestive tract microscopy), bladder (bladder microscopy), abdominal cavity (laparoscopic examination), joint cavity (arthroscopy), middle thoracic cavity (medial examination), or trachea and bronchus System (pharyngoscopy and bronchoscopy). In another embodiment, a bone marrow biopsy is performed. This type of biopsy can be performed by the sternum or vertebral body bone (the bone area on both sides of the posterior epiphysis). Clean the skin and give a local anesthetic to paralyze the area. A thin, rigid needle is inserted into the bone marrow and the cells are aspirated for study; this step occasionally causes discomfort. Central biopsy (removing small bone "shards" from the bone marrow) can follow the aspiration procedure. In another embodiment, excision or incision of a biopsy can be performed on a mammal. This type of biopsy is often used when a wider or deeper skin portion is required. Using a scalpel, remove the full skin thickness for further examination and suture the wound (using surgical sutures to suture off). When the entire tumor is removed, it is called a biopsy technique. When only a part of the swollen tumor is removed, it is called a biopsy technique. For example, when it is suspected to be a melanoma (a type of skin cancer), it is often preferred to remove the biopsy. Still other embodiments of the towel can be used to examine the fine needle neat (F N A) living tissue. This type of biopsy uses a fine needle to remove small pieces of tissue from the tumor. Occasionally, local anesthesia is used to paralyze the area, but this procedure is rare. 52 200835507 Major discomfort and no blistering. For example, FNA is not used for the examination of suspected cancer, but it can be used to examine large lymph nodes near melanoma to see if melanoma has metastasized (diffused). Computed tomography (CT scan or CAT scan) can be used to introduce fine needles into tumors of internal organs such as the lungs or liver. 5 In other embodiments, punching, shaving, and/or skin biopsy can be performed. Puncture biopsy involves the use of a biopsy instrument to remove a short cylinder or tissue called "Apple Heart" for deeper skin samples. After the local anesthesia is applied, the instrument is rotated on the surface of the skin until the instrument passes through the layers, including the dermis, epidermis, and the superficial portion of the skin (subcutaneous fat). Shaving the Living Group 10 Weaving inspection involves removing the top layer of the skin by scraping. Shaving biopsy is also performed using local anesthesia. Skin biopsy involves removing the skin sample and examining it under the microscope to determine if melanoma is present. The biopsy is performed under local anesthesia. In a specific embodiment, a method of determining whether a tumor overexpresses Akt kinase 15 is provided. The overexpression of Akt kinase can be referred to the phosphorylation state of the kinase. The high degree of squaring of Akt can be detected according to the methods described herein. In one embodiment, the tumor biopsy can be compared to control tissue. The control tissue may be a normal tissue from a mammal undergoing a biopsy or a normal tissue obtained from a healthy mammal. Overexpression of Akt kinase or high tartar 20 can be used to determine that tumor biopsy contains a greater amount of Akt kinase and/or Akt kinase squaring than the control tissue, such as at least about 1.5, 2, 2.25, compared to the Akt kinase contained in the control tissue. 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75 '5, 5.5, 6, 7, 8, 9, or 1〇 times a large number

Akt激酶。 53 200835507 於一個實施例中，本發明提供一種於個體或於得自該個體之生物樣本中檢測失序的Akt激酶表現之方法，該方法係經由讓得自該個體之細胞、細胞萃取物、血清或其它樣本或讓該生物樣本與對該Akt激酶或其抗原部分為特異性 5 之免疫交互作用分子接觸，以及篩檢免疫交互作用分子 _Akt激酶複體的形成程度，其中複體存在量相對於正常細胞升高，指示可表現或過度表現Akt之失序細胞。於一個實例中，可藉免疫學篩檢細胞或細胞萃取物是否存在有升高含量之Akt激酶。 10 於另一個實施例中，經由篩檢編碼Akt激酶基因之表現程度，來於基因層面檢測細胞中之Akt之失序表現，其中轉錄表現產物（亦即mRNA)比較正常細胞之濃度升高，係指示失序細胞。於若干實施例中，即時PCR及其它PCR程序可用來判定轉錄活性。於一個實施例中，mRNA可得自個體之 15 細胞或得自個體之生物樣本，視需要可產生cDNA。然後 mRNA或cDNA與可雜交至及/或擴增編碼Akt激酶之全部或部分核苷酸序列或其互補核苷酸序列之基因探針接觸，然後檢測mRNA或cDNA之含量，其中可評估mRNA或cDNA 之濃度是否比較正常對照組升高。 2〇本發明之又另一個實施例涵蓋於定量或半定量診斷套件組中之抗Akt激酶之抗體，包括單株抗體或多株抗體用來判定得自病人之可疑癌細胞中之Akt激酶的相對濃度，包括進行該檢定分析所需之全部試劑。於一個實施例中，提供利用施行ELISA檢定分析所需之試劑及材料之套件組。試 54 200835507 劑可包括例如洗滌緩衝液、抗體稀釋緩衝液、阻斷緩衝液、細胞染色溶液、展開溶液、停止溶液、抗磷酸-蛋白質特異性抗體、抗-泛蛋白特異性抗體、二次抗體及蒸餾水。套件組也包括使用指示’視需要可為自動化或半自動化，或呈 5 與自動化機器或軟體可相容之形式。於一個實施例中，檢測AKT活化形式（於絲胺酸474磷酸化之Akt)之磷酸-ser-473 Akt抗體可用作為診斷套件組中之抗體。例如參考Yuan等人，致癌基因，2000. 19: ρ·2324-2330。 5.6 Akt激酶 10 Akt也定名為PKB3，表示絲胺酸/蘇胺酸激酶之一個亞族。本亞族中已經識別三個成員AKT1、AKT2、及AKT3。 Akt係以PI3K-相依性方式藉胞外刺激活化（Datta S.R.等人，基因Dev，1999. 13: ρ· 2905-2927)。Akt之全活化要求活化回路中之Thr·的磷酸化及於C端活化功能部位的 15 Ser473。Akt藉PTEN腫瘤阻遏子負向調節。PTEN之突變已經於多種腫瘤中識別，結果導致Akt徑路的活化(Datta S.R· 等人，基因Dev，1999· 13: ρ·2905-2927)。此外，於多種人惡性疾病已經檢測出Akt之擴增、過度表現及/或活化(Datta S.R.等人，基因 Dev，1999. 13: ρ·2905-2927 ; Cheng J.Q·及 2〇 Nicosia S.V·，於癌症百科參考，2001，Schwab D.(編輯） Springer ’柏林、海德堡及紐約，35-37頁）。Akt特別為組成活性Akt之異位表現，誘導細胞存活及惡性轉形；而Akt活性之抑制於哺乳動物細胞之範圍内誘導細胞凋亡（Datta S.R.等人，基因 Dev，1999. 13: ρ·2905-2927 ; Cheng J.Q·及 55 200835507Akt kinase. 53 200835507 In one embodiment, the invention provides a method of detecting a disordered Akt kinase expression in an individual or in a biological sample obtained from the individual, by administering cells, cell extracts, serum from the individual Or other sample or contact the biological sample with an immunological interaction molecule specific for the Akt kinase or antigenic portion thereof, and screening for the degree of formation of the immunological interaction molecule _Akt kinase complex, wherein the amount of the complex is relatively As normal cells rise, indicating out-of-order cells that can express or overexpress Akt. In one example, the cells or cell extracts can be screened for the presence of elevated levels of Akt kinase by immunology. In another embodiment, the degree of expression of the Akt kinase gene is screened to detect the disordered expression of Akt in the cell at the genetic level, wherein the transcriptional expression product (ie, mRNA) is elevated in comparison with normal cells. Indicates out of order cells. In several embodiments, real-time PCR and other PCR programs can be used to determine transcriptional activity. In one embodiment, the mRNA can be obtained from an individual 15 cells or a biological sample obtained from an individual, which can produce cDNA as desired. The mRNA or cDNA is then contacted with a gene probe that can hybridize to and/or amplify all or part of the nucleotide sequence encoding the Akt kinase or its complementary nucleotide sequence, and then detect the mRNA or cDNA content, wherein the mRNA or cDNA can be assessed Whether the concentration of cDNA is higher than that of the normal control group. 2. Still another embodiment of the present invention encompasses antibodies against Akt kinase in a quantitative or semi-quantitative diagnostic kit, including monoclonal antibodies or polyclonal antibodies for determining Akt kinase in a suspected cancer cell from a patient. Relative concentrations, including all reagents required to perform the assay. In one embodiment, a kit of kits and materials required for performing an ELISA assay is provided. Test 54 200835507 agents may include, for example, washing buffer, antibody dilution buffer, blocking buffer, cell staining solution, developing solution, stopping solution, anti-phospho-protein specific antibody, anti-ubiquitin specific antibody, secondary antibody And distilled water. The kit group also includes instructions for use 'automatic or semi-automated as needed, or 5 compatible with automated machines or software. In one embodiment, a phospho-ser-473 Akt antibody that detects an activated form of AKT (Akt phosphorylated at serine 474) can be used as an antibody in the diagnostic kit set. See, for example, Yuan et al., Oncogene, 2000. 19: ρ·2324-2330. 5.6 Akt kinase 10 Akt is also designated PKB3 and represents a subfamily of serine/threonine kinases. Three members AKT1, AKT2, and AKT3 have been identified in this subfamily. Akt is activated by extracellular stimuli in a PI3K-dependent manner (Datta S. R. et al., Gene Dev, 1999. 13: ρ 2905-2927). The full activation of Akt requires phosphorylation of Thr· in the activation loop and 15 Ser473 at the C-terminal activation site. Akt is negatively regulated by the PTEN tumor repressor. Mutations in PTEN have been identified in a variety of tumors, resulting in activation of the Akt pathway (Datta S. R. et al., Gene Dev, 1999 13: ρ. 2905-2927). In addition, amplification, overexpression and/or activation of Akt has been detected in a variety of human malignancies (Datta SR et al., Gene Dev, 1999. 13: ρ·2905-2927; Cheng JQ· and 2〇 Nicosia SV·, Reference to Cancer Encyclopedia, 2001, Schwab D. (ed.) Springer 'Berlin, Heidelberg and New York, pp. 35-37). Akt specifically ectopically expresses active Akt, induces cell survival and malignant transformation; whereas inhibition of Akt activity induces apoptosis in mammalian cells (Datta SR et al., Gene Dev, 1999. 13: ρ· 2905-2927 ; Cheng JQ· and 55 200835507

Nicosia S.V·，於癌症百科參考，2001，Schwab D.(編輯） Springer，柏林、海德堡及紐約；35-37頁；Sun Μ·等人， Am J Path，2001· 159: ρ·431-437; Cheng J.Q·等人，致癌基因，1997· 14: p.2793-2801)。此外，Akt之活化顯示與腫瘤 5 的入侵及化學品抗性相關（West K.A.等人，藥物Resist Updat，2002. 5: ρ·234-248)。Nicosia SV·, Reference for Cancer Encyclopedia, 2001, Schwab D. (ed.) Springer, Berlin, Heidelberg, and New York; 35-37; Sun Μ· et al, Am J Path, 2001· 159: ρ·431-437; Cheng JQ et al., Oncogene, 1997·14: p. 2793-2801). Furthermore, activation of Akt has been shown to be associated with tumor 5 invasion and chemical resistance (West K.A. et al., Drug Resist Updat, 2002. 5: ρ. 234-248).

Akt徑路之活化經由誘導細胞存活、生長、遷移及血管新生，而於惡性轉形及化學品抗性之中扮演關鍵性角色。本發明提供測定Akt激酶過度表現程度及/或超活化及磷酸 10 化Akt激酶之程度之方法。Activation of the Akt pathway plays a key role in malignant transformation and chemical resistance by inducing cell survival, growth, migration, and angiogenesis. The present invention provides methods for determining the extent of Akt kinase overexpression and/or the degree of hyperactivation and phosphorylation of Akt kinase.

Akt激酶可為任一種已知之Akt家族激酶，或其相關激酶，包括但非限於Aktl、Akt2、Akt3。人Aktl、Akt2、及 Akt3之mRNA及胺基酸序列分別顯示於第6a-c、7a-(L〇a-c 圖。 15 5·7·診斷檢定分析免疫檢定分析於一個實施例中，提供一種於哺乳動物細胞或於得自哺乳動物之生物樣本檢測Akt激酶之失序表現之方法，該方法係經由讓得自該哺乳動物之細胞、細胞萃取物或血清或 20其匕樣本或生物樣本與對Akt激晦或其抗原部分具有特異性之免疫交互作用分子接觸；以及篩檢免疫交互作用分子 -Akt激酶複體形成程度；以及判定是否存在有相對於正常細胞複體存在量之升高。免疫交互作用分子可為對Akt激酶或其抗原部分或其 56 200835507 同系物或何生物具有特異性及結合親和力。於一個實施例中，免疫交互作用分子可為免疫球蛋白分子。於其它實施例中，免疫交互作用分子可為抗體片段、單鏈抗體及/或去免疫化分子包括人化抗體及抗原結合分子相關之τ細胞 5 (键二)。於一特定實施例中，抗體可為單株抗體。於另一寺疋貝施例中抗體可為多株抗體。免疫交互作用分子對 Akt激酶有特異性，或更特別對恤激酶上的抗原定子或抗原決定部位有特異性。他激酶上的抗原定子或抗原決定部位包括免疫反應所針對的分子部分。抗原定子或抗原決定 10 W位可為B細胞抗原決定部位，或若屬適當為了細胞抗原決定部位。於一個實施例中，抗體為磷酸-ser 473 Akt抗體。本發明之一個實施例提供一種於哺乳動物其中存在有失序Akt活性之癌症生長或癌症系列生長之存在之方法，係經由讓得自該哺乳動物之細胞或細胞萃取物或得自個體之生物樣本與Akt激酶結合有效量之具有Akt激酶特異性之抗體或其上之抗原定子或抗原決定部位接觸；以及然後定量或定性測定Akt激酶-抗體複體之含量，其中判定該複體比較正常細胞存在有較高含量。抗體可藉熟諳技藝人士已知之多種手段中之任一者製 2〇備。例如用於人Akt激酶之檢測，抗體通常但非必要衍生自非人動物諸如靈長類、牲口動物(例如羊、牛、豬、山羊、馬）、實驗室試驗動物（例如小鼠、大鼠、天竺鼠、兔)及/或伴但動物(例如犬、貓）。抗體也可於原核宿主細胞或真核宿主細胞以重組方式製造。大致上，基於抗體之檢定分析可 57 200835507 於試管内於細胞或組織之生檢進行。但若抗體經過適當脫免疫化，或於供人類使用之情況下，抗體經過人化，則該抗體例如可以細胞核標籤標記，投予病人，藉放射性技術來測定核標記堆積位置。Akt激酶抗體可為癌症乾定劑。如 5此，本發明之另一個實施例提供用於人類及非人病人之癌症成像用抗體之脫免疫化形式。大致上，為了產生抗Akt激酶之抗體，要求酶係萃取自生物樣本，而生物樣本可來自於動物包括人組織，或若係藉重組手段製造則可來自於細胞培養。Akt激酶可藉任一種 10適當手段而分離自生物樣本。例如分離可利用Akt激酶表面電荷性質、大小、密度、生物活性及對另一個實體(例如Akt 激酶所結合或以其它方式相關聯之另一個蛋白質或化學化合物）之親和力中之任一者或任多者。如此，例如，由生物流體分離Akt激酶可藉下列方法中之任一者或任多者來達 15成：超離心、離子交換層析術(例如陰離子交換層析術、陽離子父換層析術）、電泳(例如聚丙稀醯胺凝膠電泳、等電聚焦）、尺寸分離(例如凝膠過濾、超濾)及親和力媒介分離(例如免疫親和力分離包括但非限於磁珠分離諸如戴納珠 (Dynabead)(商品名）分離、免疫層析術、免疫沉殿）。Akt 20激酶由生物流體之分離可保有存在於激酶上之隨形抗原決定部位，如此適當避開造成酶變性之技術。於又一實施例中，可使用親和分離、凝膠過濾及/或超濾中之任一者或多者而由生物流體分離激酶。免疫接種以及隨後單株抗體之製造可使用技藝界已知 58 200835507 之任一種標準協定方案來進行，例如說明於K〇hler及The Akt kinase can be any of the known Akt family kinases, or their related kinases, including but not limited to Aktl, Akt2, Akt3. The mRNA and amino acid sequences of human Aktl, Akt2, and Akt3 are shown in Figures 6a-c, 7a-(L〇ac map, respectively. 15 5·7·Diagnostic assay analysis Immunoassay analysis in one embodiment provides one A method for detecting the disordered expression of Akt kinase in a mammalian cell or a biological sample obtained from a mammal by administering cells, cell extracts or serum derived from the mammal or 20 sputum samples or biological samples to Akt The stimulating or antigenic portion has specific immunological interactions with molecular contacts; and screening for immunological interaction molecules - the extent of Akt kinase complex formation; and determining whether there is an increase in the amount of presence relative to normal cell complexes. The acting molecule can be specific for and binding affinity to Akt kinase or an antigenic portion thereof or its 56 200835507 homolog or organism. In one embodiment, the immunologically interacting molecule can be an immunoglobulin molecule. In other embodiments, The immunological interaction molecule can be an antibody fragment, a single-chain antibody, and/or a de-immunized molecule including a humanized antibody and an antigen-binding molecule associated with τ 5 (key 2). In a specific embodiment, the antibody may be a monoclonal antibody. In another case, the antibody may be a plurality of antibodies. The immunological interaction molecule is specific to Akt kinase, or more specific. Specificity of the antigenic stator or epitope on the kinase kinase. The antigenic stator or epitope of the kinase includes the molecular portion to which the immune response is directed. The antigenic stator or antigen determines that the 10 W position can be a B cell epitope. Or, if appropriate, for a cellular epitope. In one embodiment, the antibody is a phospho-ser 473 Akt antibody. One embodiment of the invention provides a cancer growth or cancer series growth in mammals in which disorder Akt activity is present. The method of presenting is by contacting an Akt kinase-binding antibody or an antigenic stator or epitope thereof with an Akt kinase-binding amount of a cell or cell extract obtained from the mammal or a biological sample obtained from the individual. And then quantitatively or qualitatively determining the amount of Akt kinase-antibody complex, wherein it is determined that the complex is present in comparison with normal cells. High levels of antibodies can be prepared by any of a variety of means known to those skilled in the art. For example, for the detection of human Akt kinases, antibodies are usually, but not necessarily, derived from non-human animals such as primates, livestock animals ( For example, sheep, cattle, pigs, goats, horses, laboratory test animals (such as mice, rats, guinea pigs, rabbits) and / or accompanying animals (such as dogs, cats). Antibodies can also be in prokaryotic host cells or true Nuclear host cells are produced recombinantly. In general, antibody-based assays can be performed in vitro on cells or tissues in vitro, but if the antibodies are properly de-immunized or used in humans, antibodies After humanization, the antibody can be labeled, for example, by a nuclear label, and administered to a patient to determine the location of the nuclear marker by radioactive techniques. The Akt kinase antibody can be a cancer dry agent. As a fifth, another embodiment of the present invention provides a de-immunized form of an antibody for cancer imaging for use in humans and non-human patients. In general, in order to produce an antibody against Akt kinase, the enzyme is required to be extracted from a biological sample, and the biological sample may be derived from an animal including human tissue, or may be derived from cell culture if it is produced by recombinant means. Akt kinase can be isolated from biological samples by any suitable means. For example, isolation may utilize any of the Akt kinase surface charge properties, size, density, biological activity, and affinity for another entity (eg, another protein or chemical compound that is associated or otherwise associated with Akt kinase) More. Thus, for example, separation of Akt kinase from a biological fluid can be achieved by any one or more of the following methods: ultracentrifugation, ion exchange chromatography (eg, anion exchange chromatography, cation-parent tomography) ), electrophoresis (eg, polyacrylamide gel electrophoresis, isoelectric focusing), size separation (eg, gel filtration, ultrafiltration), and affinity media separation (eg, immunoaffinity separation including, but not limited to, magnetic bead separation such as Dyna beads ( Dynabead) (trade name) separation, immunochromatography, immunosuppression hall). The separation of the Akt 20 kinase from the biological fluid preserves the conformational epitope of the conformation present on the kinase, so that the technique responsible for enzymatic denaturation is appropriately avoided. In yet another embodiment, the kinase can be isolated from the biological fluid using any one or more of affinity separation, gel filtration, and/or ultrafiltration. Immunization and subsequent manufacture of monoclonal antibodies can be carried out using any of the standard protocol protocols known in the art 58 200835507, for example as described in K〇hler and

Milstein (Kohler及Milstein，自然，1975. 256: ρ·495·499 ; Kohler及Milstein，Eur J Iminun〇l，1976. 6(7): ρ·511-519);Milstein (Kohler and Milstein, Nature, 1975. 256: ρ·495·499; Kohler and Milstein, Eur J Iminun〇l, 1976. 6(7): ρ·511-519);

Coligan等人（免疫學之流行方案，^9^997，約翰威利父 5 子公司）或T〇yama等人（單株抗體實驗手冊，1987年，科學講談社）。大致上，動物係以含Akt激酶之生物流體或其部为或Akt激酶之重組形式精標準方法免疫接種，來產生可製造抗體之細胞，特別為抗體製造性體細胞（例如B淋巴細胞）。此等細胞隨後由免疫接種動物體分離進行永生處理。 10於若干實施例中，Akt激酶片段可用來產生抗體。該片段可與一載劑連結。該載劑可為典型高分子量之任何物質，非免疫原性物質或不良免疫原性物質（例如半抗原）自然鍵聯或以人工方式鍵聯至載劑來提升其免疫原性。抗體製造性細胞之永生化可使用技藝界眾所周知之方 15法進行。例如經由使用EB病毒(EBV)之轉形方法可達成永生化(Kozbor等人，酶學方法，1986· 121: ρ·ΐ4〇)。於另一個實施例中，抗體製造性細胞係使用細胞融合法來永生化(述於Coligan等人，199Μ997，參見上文），細胞融合法廣用於製造單株抗體。於此種方法中，具有潛力製造抗體之抗 20體製造性體細胞，特別為B細胞係與骨髓瘤細胞系融合。2 等體細胞可衍生自二次感染動物較佳為誓齒類動物諸如小鼠及大鼠之淋巴結、脾臟及及周邊血液。於特定實施例中可使用小鼠脾細胞。於其它實施例中，也可使用大鼠、兔羊或山羊細胞。由淋巴細胞性腫瘤已經發展出特化骨骑瘤 59 200835507 細胞來用於融合瘤製造之融合程序（Kohler及Milstein， 1976,參見上文；Shulman等人，自然，1978.276: ρ·269-270 ; Volk等人，J Virol，1982· 42(1): ρ·220-227)。多種骨髓瘤細胞系也可用於融合細胞雜交體之製造，包括例如P3乘 5 63-Ag8、P3 乘 63-AG8.653、P3/NSl-Ag4-l(NS-l)、Coligan et al. (Popular Program for Immunology, ^9^997, John Wiley's Parent 5 Subsidiary) or T〇yama et al. (Single Antibody Laboratory Manual, 1987, Science Lecture). In general, animals are immunized with a biological fluid containing Akt kinase or a recombinant form thereof, or a recombinant form of Akt kinase, to produce cells capable of producing antibodies, particularly antibody-producing somatic cells (e.g., B lymphocytes). These cells are then separated from the immunized animal for immortalization. In several embodiments, Akt kinase fragments can be used to produce antibodies. This fragment can be linked to a carrier. The carrier can be any material of a typical high molecular weight, non-immunogenic or poorly immunogenic (e.g., hapten) naturally linked or artificially linked to the carrier to enhance its immunogenicity. Immortalization of antibody-producing cells can be carried out using methods well known in the art. For example, Kombor et al., Enzymology Method, 1986. 121: ρ·ΐ4〇 can be achieved by a transformation method using Epstein-Barr virus (EBV). In another embodiment, antibody-producing cell lines are immortalized using cell fusion methods (described in Coligan et al., 199, 997, supra), and cell fusion methods are widely used to produce monoclonal antibodies. In this method, an anti-20-body-producing somatic cell having the potential to produce an antibody, particularly a B-cell line and a myeloma cell line, is fused. 2 The somatic cells may be derived from the secondary infected animal, preferably the vagina, such as the lymph nodes, spleen and peripheral blood of the mouse and rat. Mouse spleen cells can be used in certain embodiments. In other embodiments, rat, rabbit or goat cells can also be used. Specialized bone riding tumors have been developed from lymphocytic tumors to produce a fusion procedure for fusion tumor production (Kohler and Milstein, 1976, see above; Shulman et al., Nature, 1978.276: ρ·269-270; Volk et al., J Virol, 1982. 42(1): ρ·220-227). A variety of myeloma cell lines can also be used in the manufacture of fusion cell hybrids, including, for example, P3 by 5 63-Ag8, P3 by 63-AG8.653, P3/NSl-Ag4-l (NS-l),

Sp2/0-Agl4及S194/5.XXO.Bu.l。P3乘63-Ag8及NS-1 細胞系已經由Kohler及Milstein說明（1976，參見上文）。Shulman等人（1978，參見上文）發展Sp2/0-Agl4骨髓瘤細胞系。 S194/5.XXO.Bu.l 細胞系係由 Trowbridge報告（J Exp Med， 10 1978. 148(1): ρ·313-323)。產生抗體製造性脾細胞或淋巴結細胞與骨髓瘤細胞雜交體之方法通常涉及於可促進細胞膜之融合之作用劑（化學、病毒或電氣)存在下，將體細胞與骨髓瘤細胞以10:1比例混合（但該比例可由約20:1變化至約 1:1)。已經說明融合方法（Kohler及Milstein，1975,參見上 15 文；Kohler及Milstein，1976，參見上文；Gefter等人，體細胞遺傳學，1977· 3: p.231-236 ; Volk等人，1982，參見上文）。該等研究學者所使用之融合促進劑為山戴病毒(Sendai virus)及聚乙二醇(PEG)。於若干實施例中，提供選擇得自其餘未融合細胞，特別為未融合骨髓瘤細胞之融合細胞雜 2〇父體之手段。大致上，融合細胞雜交體之選擇伴隨有於培養基中培養細胞’該培養基可支援融合瘤的生長’但阻止未融合骨髓瘤細胞之生長，未融合骨髓瘤細胞通常會繼續無限***。融合所使用之體細胞於試管内培養旅未保有長期存活力，如此不會成問題。需要數週來選擇性培養融合 60 200835507 細胞雜交體。於本時間早期，需要識別可製造期望之抗體之該等雜交體’讓其隨後可被轉殖及繁殖。大致上，約10% 所得雜交體可製造期望之抗體，但約1%至約30%之範圍並非不常見。抗體製造性雜交體之檢測可藉數種標準檢定分 5 析方法中之任一種達成，該等方法包括酶聯結免疫檢定分析技術及放射性免疫檢定分析技術，例如說明於Kennet等人(單株抗體及融合瘤··生物分析之新紀元，1980年，普列能出版社(Plenum Press)，紐約，376-384頁）；且可藉FACS 分析達成(O’Reilly等人，生物技術，1998 25: p.824-830)。 10 —旦已經選擇期望之融合細胞雜交體且轉殖入個別抗體製造性細胞糸’各細胞系可於兩種標準方式之任一種繁殖。融合瘤細胞懸浮液注射組織可相容的動物體内。被注射的動物隨後發展出可分泌由該融合細胞雜交體所製造之特定單株抗體。動物體液諸如血清或腹水可經輕敲來以高 15 濃度製造單株抗體。另外，個別細胞系可於試管内於實驗室培養容器中繁殖。含高濃度單一特定單株抗體之培養基可藉傾析、過濾或離心收穫，以及隨後純化。然後細胞系藉免疫檢測手段測試其檢測感興趣之A k t 激酶之特異性。舉例言之，細胞系可分配入多個孔内及培 20 養’來自各孔之上清液藉酶聯結免疫吸附檢定分析 (EUSA)、間接螢光抗體技術等分析。可辨識標靶LIM激酶但不會辨識非標靶抗原決定部位之單株抗體製造性細胞系經過識別，然後於試管内直接培養，或注射入組織可相容動物體内來形成腫瘤，以及製造、收集、及純化所需抗 61 200835507Sp2/0-Agl4 and S194/5.XXO.Bu.l. The P3 by 63-Ag8 and NS-1 cell lines have been described by Kohler and Milstein (1976, see above). Shulman et al. (1978, supra) developed the Sp2/0-Agl4 myeloma cell line. The S194/5.XXO.Bu.l cell line is reported by Trowbridge (J Exp Med, 10 1978. 148(1): ρ·313-323). A method for producing antibody-producing spleen cells or a mixture of lymph node cells and myeloma cells is generally directed to a 10:1 ratio of somatic cells to myeloma cells in the presence of an agent (chemical, viral or electrical) that promotes fusion of cell membranes. Mixing (but the ratio can vary from about 20:1 to about 1:1). Fusion methods have been described (Kohler and Milstein, 1975, see above 15; Kohler and Milstein, 1976, supra; Gefter et al, Somatic Genetics, 1977. 3: p. 231-236; Volk et al., 1982 , see above). The fusion promoters used by these researchers are Sendai virus and polyethylene glycol (PEG). In several embodiments, a means for selecting a fused cell heterozygote derived from the remaining unfused cells, particularly unfused myeloma cells, is provided. In general, the selection of fused cell hybrids is accompanied by the culture of cells in the medium [this medium supports the growth of fusion tumors] but prevents the growth of unfused myeloma cells, which usually continue to divide indefinitely. The in vivo culture of the somatic cells used for fusion does not retain long-term viability, so this is not a problem. It takes several weeks to selectively culture the fusion 60 200835507 cell hybrid. Early in the day, it is necessary to identify such hybrids that can produce the desired antibodies' so that they can subsequently be propagated and propagated. In general, about 10% of the resulting hybrids produce the desired antibodies, but ranges from about 1% to about 30% are not uncommon. The detection of antibody-producing hybrids can be achieved by any of several standard assays, including enzyme-linked immunoassay techniques and radioimmunoassay techniques, as described, for example, in Kennet et al. And a new era of fusion tumors, bioanalysis, Plenum Press, New York, pp. 376-384, 1980; and can be reached by FACS analysis (O'Reilly et al., Biotechnology, 1998 25: P.824-830). 10 Once the desired fused cell hybrid has been selected and transfected into individual antibody-producing cells, each cell line can be propagated in either of two standard ways. The fusion tumor cell suspension is injected into a tissue compatible animal. The injected animal is then developed to secrete specific monoclonal antibodies produced by the fused cell hybrid. Animal body fluids such as serum or ascites can be tapped to produce monoclonal antibodies at high concentrations of 15 . Alternatively, individual cell lines can be propagated in vitro in a laboratory culture vessel. A medium containing a high concentration of a single specific monoclonal antibody can be harvested by decantation, filtration or centrifugation, and subsequently purified. The cell line is then tested by immunoassay for its specificity for detecting the A k t kinase of interest. For example, the cell line can be dispensed into a plurality of wells and cultured from each well by enzyme-linked immunosorbent assay (EUSA), indirect fluorescent antibody technique, and the like. Individual antibody-producing cell lines that recognize the target LIM kinase but do not recognize non-target epitopes are identified, then cultured directly in vitro, or injected into tissue-compatible animals to form tumors, and manufactured , collection, and purification of the required anti-61 200835507

因此，本發明提供一種於一樣本中檢測Akt激酶或其片段、變異株或衍生物之方法，包含該樣本與抗體或其片段或其衍生物接觸；檢測比較正常對照組，含有該抗體及Akt 5激酶或其片段變異株或衍生物之複體之含量，其中測定Akt 激酶之升局程度。任何測定複體之形成之適當技術皆可使用。舉例g之，根據本發明具有結合之通報子分子之抗體可用於免疫檢定分析。此種免疫檢定分析包括但非限於放射性免疫檢定分析（RIA)、酶聯結免疫吸附檢定分析 10 (ELISA)、免疫層析技術(ICT)及西方墨點，此等免疫檢定分析為熟諳技藝人士眾所周知。免疫檢定分析也包括競爭檢定分析。本發明涵蓋定性及定量免疫檢定分析。適^免疫檢定分析技術例如述於美國專利案 4,016,043，4,424,279 ;及4，G18,653。此等技術包括非競爭 I5型單位置及二位置檢定分析及傳統競爭結合檢定分析。此專才欢疋刀析包括經過標§己之抗原結合分子直接結合至標把抗原。本發明進一步提供於得自動物體諸如癌症病人或懷疑患有癌症個體之細胞或組織樣本中定量Akt蛋白質表現程 2〇度及活化程度之方法。於一個實施例中，本發明提供定量使用成像系統來定量Akt蛋白質表現或活化程度之方法。成像系統可用來接收、提升及處理已經使用AKT蛋白質特異性染色進㈣色之細胞樣本或組織樣本之影像，俾便判定於得自此種動物之細胞或組織樣本中之丁蛋白質表現量 62 200835507 或活化程度。於本發明方法之一個實施例中，可對表現不等量AKT蛋白質之至少兩個細胞系產生八纽及akt2蛋白質表現校準曲線。然後校準曲線用來定量測定於細胞或組織樣本中表現之ΑΚΤ蛋白質數量。可使用活化特徵特異性 5试劑來對活化ΑΚΤ蛋白質製作類似的校準曲線。也可用來判定於臨床癌症治療前與治療後之ΑΚΤ數量及ακτ活化狀態的變化。 ;本毛月方/irt自特定實施例中，於細胞或組織樣本中之AKT蛋白質表現程度可使用酶聯結免疫吸附檢定分 1〇析(EUSA)定量’來測定樣本中之AKT蛋白質含量。此種方法例如述於美國專利公告案2〇〇2/〇〇丨MM。於其它實施例中’ _免疫檢定分析可用來檢測Akt激酶。於此種檢定分析中，酶輛合至第二抗體，通常係利用戍二駿或過峨酸鹽來車厄合至第二抗體。用於特定酶之酶基 15質通常係於被相對應之崎水解時產生可檢測之顏色變化而選出。也可使用獲得螢光產物之螢光產生性酶基質，而非使用產色酶基質。經過崎標記之抗體可添加至第—抗體-抗原複體，允許其結合，然後洗務去除過量試劑。含有適當酶基質之溶液隨後添加至抗體-抗原.抗體複體。酶基質與聯 20結至第二抗體之酶反應，產生定量視覺信號，該信號通常可糟分光光度辦法進-步定量，來獲得樣本中所存在之抗原里之指不。另外，螢光化合物諸如榮光素、若丹明、及鋼系化合物、銪(EU)可化學偶合至抗體而未變更其結合能力。當藉特定波長之光照明而活化時，經過榮光絡標記之 63 200835507 抗體吸收光能，於分子内誘導呈激發狀態，接著發出通常使用光學顯微鏡可視覺檢測之特定色彩之光。經過螢光標記之抗體讓其結合至第一抗體-抗原複體。於洗滌去除未結合之反應劑之後，其餘三元體複合體暴露於適當波長之 5 光。觀察得螢光指示存在有感興趣之抗原。免疫螢光計量檢定分析(IFMA)於技藝界已經明確確立，特別可用於本方法。但也可採用其它通報子分子諸如放射性同位素、化學發光分子或生物發光分子。於特定實施例中，Akt激酶之抗體也可用於Akt激酶特 10別血清或其它循環體液中之Akt激酶藉ELISA媒介之檢測。可經由將抗-Akt激峰抗體制動於固體樓體，且讓其與生物萃取物諸如血清、血液、淋巴或其它體液、細胞萃取物或細胞生檢接觸來達成。然後經過標記之抗Akt激酶抗體可用來檢測經過制動之Akt激酶。本檢定分析可以多種方式 15之任一種檢測，全部變化皆涵蓋於本發明且為熟諳技藝人士已知。此項辦法允許例如使用基於血清之檢定分析來快速檢測與量化Akt激酶含量。於一個實施例中Akt ELISA檢定分析套件組可用於本發明。例如得自超陣列生科公司（SuperArray Bi〇sdence)之 20 Akt S473用之發訊細胞活化ELISA套件組可用於本發明。於一個實施例中，抗體可為識別Akt S473之抗_泛抗體。EUSA 檢定分析套件組含有抗-Akt抗體及額外試劑，包括但非限於洗滌緩衝液、抗體稀釋緩衝液、阻斷緩衝液、細胞染色溶液、展開〉谷液、停止溶液、二次抗體及蒸餾水。 64 200835507 核苷酸檢測於另一個實施例中，提供經由檢測編碼Akt激酶之多核苷酸於細胞中之表現程度來檢測Akt激酶之方法。多核苷酸之表現可使用热諸技藝人士已知之任一種適當技術測定。 5於一個實她例中’編碼Akt激酶之經過標記之多核苷酸可用作為探針用於得自細胞之RNA萃取物之北方墨點分析。於其它實施例中’得自動物之核酸萃取物可與與編碼該激酶之多核甘酸之訊息序列及反訊息序列或由其中之側支序列相對應之券核誓酸引子協同用於核酸擴增反應，諸如RT 10 PCR。多種自動化固相檢測技術也為熟諳技藝人士可利用，例如述於Fodor等人(科學，1991· 251·· ρ·767-777)及Kazal 等人（自然藥物，1996. 2: p.753-759)。於其它實施例中，提供檢測編碼Akt激酶之RNA轉錄本之方法。RNA可分離自懷疑含有Akt激酶rnA之細胞樣本， 15 例如分離自人癌組織之全RNA。RNA可藉技藝界已知之方法例如使用TRIZ0L試劑(GIBC0-BRL/生命科學，馬里蘭州蓋色堡）分離。寡-dT或隨機序列寡核苷酸及特定序列寡核苷酸可用作為反錄酶反應中作為引子，來製備得自經分離之RNA之第一股cDNA。然後所得第一股cDNA於PCR反應 2〇中以特定序列募核苔酸擴增來獲得擴增產物。聚合酶連鎖反應或「PCR」係指其中核酸、RNA及/或 DNA之預選定片段之數量經擴增之程序或技術，例如說明於美國專利案4,683，195。大致上，得自感興趣區末端或超越該區之序列資訊用來設計寡核苷酸引子。此等引子之序 65 200835507 列可與欲擴增之樣板相對股之序列相同或類似。可使用 PCR來擴增特定RNA序列及由全細胞RNA轉錄之cDNA。大致上參考Mullis等人(Quant Biol，1987· 51: ρ·263; Erlich編輯，PCR技術，1989，史塔克敦出版社，紐約）。如此，藉 5 PCR擴增特定核酸序列仰賴具有保留的核苷酸序列之寡核苷酸或引子，其中該保留序列係由相關基因序列或蛋白質序列之校準推定，例如由哺乳動物Akt激酶基因之序列比較推定。舉例言之，製備一個引子，該引子預測可與反訊息股配對；以及製備另一個引子，該引子預測可與編碼Akt 10 激酶之cDNA分子之訊息股配對。欲檢測經擴增之產物，反應混合物典型接受瓊脂糖凝膠電泳或其它方便的分離技術，檢測經過Akt激酶特異性擴增DNA之相對存在。例如，經過Akt激酶擴增之DNA可使用與特定寡核苔酸探針之南方雜交檢測，或將其電泳活動性與具有已知分子量之DNA 15 標準比較。經擴增之Akt激酶DNA之分離、純化及特徵化可經由由凝膠中切除該片段或洗提該片段（例如參考參考文獻Lawn等人，核酸研究，1981. 2: ρ· 6103; Goeddel等人，核酸研究，1980· 8: ρ·4057);將擴增所得產物轉殖入適當載體諸如pCRII載體（因維左金公司（jnvitr〇gen))之轉殖位置； 2〇將轉殖後之插子定序；且將該DNA序列與LIM激酶之已知序列比較來達成。然後測定LIM激酶mRNA及cDNA之相對量。於一個實施例中，即時PCR用來測定Akt核苷酸之轉錄程度。轉錄活性之判定也包括基於可用mRNA轉錄本之可 66 200835507 能轉譯活性之測定。即時PCR程序及其它PCR程序使用多種 PCR產物之檢測化學，包括DNA結合螢光基團之結合、5, 核酸内切酶、相鄰内襯及髮夾募探針及自行發螢光擴增基因。此等化學及即時PCR大致上係討論於Mackay等人，核 5 酸研究 ’ 2002· 30(6): p.1292-1305; Walker，J Biochem Mol 母理學 ’ 2001. 15(3): ρ.121_127; Lewis等人，J Pathol，2001. 195: p.66-71。於另一個實施例中，Akt之失序表現可藉下述方法識別’經由讓分離自一生物樣本之核苷酸序列與具有選自於 10 第6a-c、7a-d、或8a-c圖之寡核苷g曼序列之Akt序列或其片段互補之一序列之寡核誓酸探針接觸；以及然後經由該探針與序列雜交且將結果與正常樣本比較來檢測該序列。探針與生物樣本之雜交可經由使用任一種可檢測劑來標記該探針而檢測。探針例如可以放射性同位素，或以可取得抗 15 體之生物素、螢光染料、電子稠密試劑、酶、半抗原或蛋白質標記。可檢測標記可藉任何期望之手段包括分光手段、光化學手段、生物化學手段、免疫化學手段、放射性同位素手段、或化學手段檢定分析。探針也可使用諸如寡核苷酸限剪技術、點打點檢定分析、反點打點檢定分析、 20 線探針檢定分析及5’磷酸酶檢定分析。另外，探針可使用一般適用之DNA陣列技術中之任一者來檢測，包括巨陣列技術、微陣列技術及DNA微晶片技術。寡核苔酸探針典型包括雜交至選自於第6a_c、7a_d及8a-c圖之核苷酸或其片段之約14、15、16、18、20、25或28核苷酸。通常使用長度 67 200835507 大於約25或28核苷酸之探針並不佳。寡核苷酸探針設計來識別Akt核苷酸序列。激酶檢定分析Accordingly, the present invention provides a method for detecting Akt kinase or a fragment, variant or derivative thereof, which comprises contacting the sample with an antibody or a fragment thereof or a derivative thereof; detecting a normal control group containing the antibody and Akt The content of a complex of 5 kinase or a variant thereof or a derivative thereof, wherein the degree of progression of Akt kinase is determined. Any suitable technique for determining the formation of a complex can be used. For example, an antibody having a combined reporter molecule according to the present invention can be used for immunoassay analysis. Such immunoassay analysis includes, but is not limited to, radioimmunoassay analysis (RIA), enzyme-linked immunosorbent assay (ELISA) 10, immunochromatographic (ICT), and Western blotting, which are well known to those skilled in the art. . Immunoassay analysis also includes competitive assay analysis. The invention encompasses qualitative and quantitative immunoassay analysis. Suitable immunoassay techniques are described, for example, in U.S. Patent Nos. 4,016,043, 4,424,279; and 4, G18,653. These technologies include non-competitive I5 type single position and two position verification analysis and traditional competitive combination verification analysis. This expert analysis involves the direct binding of the antigen-binding molecule to the target antigen. The invention further provides methods for quantifying Akt protein expression and degree of activation in a cell or tissue sample obtained from an automated subject such as a cancer patient or an individual suspected of having cancer. In one embodiment, the invention provides a method of quantitatively using an imaging system to quantify the extent of Akt protein expression or activation. The imaging system can be used to receive, enhance, and process images of cell samples or tissue samples that have been specifically stained with (4) proteins using AKT proteins, and to determine the amount of protein expressed in cells or tissue samples derived from such animals. 62 200835507 Or degree of activation. In one embodiment of the methods of the invention, a calibration curve for eight- and akt2 protein expression can be generated for at least two cell lines exhibiting unequal amounts of AKT protein. The calibration curve is then used to quantify the amount of sputum protein expressed in the cell or tissue sample. A similar calibration curve can be made for the activated prion protein using the Activation Feature Specific 5 Reagent. It can also be used to determine the number of sputum and the activation of ακτ before and after clinical cancer treatment. Ben Maoyue/irt From a specific example, the degree of AKT protein expression in a cell or tissue sample can be determined by enzyme-linked immunosorbent assay (EUSA) quantification to determine the AKT protein content in the sample. Such a method is described, for example, in U.S. Patent Publication No. 2〇〇2/〇〇丨MM. In other embodiments, the '-immunoassay assay can be used to detect Akt kinase. In this assay, the enzyme is coupled to a second antibody, usually by hydrazine or perrhenate to the second antibody. The enzyme base for a particular enzyme is typically selected to produce a detectable color change upon hydrolysis by the corresponding sinus. Instead of using a chromogenic enzyme matrix, a fluorescent-producing enzyme matrix that produces a fluorescent product can also be used. Saki-labeled antibodies can be added to the first antibody-antigen complex, allowed to bind, and then washed to remove excess reagent. A solution containing the appropriate enzyme matrix is then added to the antibody-antigen. antibody complex. The enzyme matrix reacts with the enzyme of the conjugated to the second antibody to produce a quantitative visual signal that is typically quantified by spectrophotometry to obtain an indication of the presence of the antigen in the sample. Further, a fluorescent compound such as glover, rhodamine, and a steel compound, ruthenium (EU) can be chemically coupled to the antibody without changing its binding ability. When activated by illumination of a specific wavelength of light, the luminescence-labeled 63 200835507 antibody absorbs light energy, induces excitation in the molecule, and then emits light of a specific color that is normally visually detectable using an optical microscope. The antibody is conjugated to the first antibody-antigen complex by a fluorescent marker. After washing to remove unbound reactants, the remaining ternary complex is exposed to 5 light of the appropriate wavelength. Fluorescence is observed to indicate the presence of an antigen of interest. Immunofluorescence metrology assays (IFMA) have been clearly established in the art world and are particularly useful in this method. However, other reporter molecules such as radioisotopes, chemiluminescent molecules or bioluminescent molecules may also be employed. In particular embodiments, antibodies to Akt kinase can also be used in Akt kinase-specific serum or other circulating body fluids for detection of Akt kinase by ELISA vectors. This can be achieved by immobilizing the anti-Akt peak antibody to a solid building and contacting it with a biological extract such as serum, blood, lymph or other body fluids, cell extracts or cell biopsy. The labeled anti-Akt kinase antibody can then be used to detect braked Akt kinase. The assay can be detected in any of a variety of ways, all of which are encompassed by the present invention and are known to those skilled in the art. This approach allows for rapid detection and quantification of Akt kinase levels, for example, using serum-based assays. In one embodiment, an Akt ELISA assay kit set can be used in the present invention. For example, a set of signaling cell activation ELISA kits for 20 Akt S473 from SuperArray Bissence can be used in the present invention. In one embodiment, the antibody can be an anti-pan-antibody that recognizes Akt S473. The EUSA assay kit kit contains anti-Akt antibodies and additional reagents including, but not limited to, wash buffer, antibody dilution buffer, blocking buffer, cell staining solution, unfolding > trough, stop solution, secondary antibody, and distilled water. 64 200835507 Nucleotide Detection In another embodiment, a method of detecting Akt kinase by detecting the extent of expression of a polynucleotide encoding Akt kinase in a cell is provided. The performance of the polynucleotide can be determined using any suitable technique known to those skilled in the art. 5 In one example, a labeled polynucleotide encoding an Akt kinase can be used as a probe for northern blot analysis of RNA extracts from cells. In other embodiments, the nucleic acid extract of the animal can be used for nucleic acid amplification in cooperation with a message sequence and an anti-message sequence encoding the polynucleotide of the kinase or a nucleoside primer corresponding to the collateral sequence therein. Reactions such as RT 10 PCR. A variety of automated solid phase detection techniques are also available to those skilled in the art, for example, as described by Fodor et al. (Science, 1991. 251··ρ·767-777) and Kazal et al. (Nature Drugs, 1996. 2: p.753- 759). In other embodiments, methods of detecting an RNA transcript encoding an Akt kinase are provided. RNA can be isolated from a sample of cells suspected of containing Akt kinase rnA, 15 such as total RNA isolated from human cancer tissue. RNA can be isolated by methods known in the art, for example using TRIZ0L reagent (GIBC0-BRL/Life Sciences, Gaiburg, MD). The oligo-dT or random sequence oligonucleotide and the specific sequence oligonucleotide can be used as a primer in the reverse enzymatic reaction to prepare the first strand of cDNA obtained from the isolated RNA. The resulting first strand of cDNA is then amplified in a specific sequence in a PCR reaction to obtain an amplification product. Polymerase chain reaction or "PCR" refers to a procedure or technique in which the number of preselected fragments of nucleic acids, RNA and/or DNA is amplified, as described, for example, in U.S. Patent No. 4,683,195. In general, sequence information from the end of the region of interest or beyond the region is used to design oligonucleotide primers. The order of these primers 65 200835507 The column may be identical or similar to the sequence of the strands to be amplified. PCR can be used to amplify specific RNA sequences and cDNA transcribed from whole cell RNA. Refer to Mullis et al. (Quant Biol, 1987 51: ρ. 263; Erlich, ed., PCR, 1989, Starkton, New York). Thus, 5 PCR amplification of a particular nucleic acid sequence relies on an oligonucleotide or primer having a retained nucleotide sequence, wherein the retention sequence is presumed by the alignment of the relevant gene sequence or protein sequence, eg, by a mammalian Akt kinase gene. The sequence comparison is presumed. For example, a primer is prepared which is predicted to be paired with an anti-information strand; and another primer is prepared which is predicted to be paired with a message strand encoding a cDNA molecule of Akt 10 kinase. To detect the amplified product, the reaction mixture is typically subjected to agarose gel electrophoresis or other convenient separation technique to detect the relative presence of Akt kinase-specific amplified DNA. For example, DNA amplified by Akt kinase can be detected by Southern hybridization with a specific oligo-sera probe or by comparing its electrophoretic activity to a DNA 15 standard of known molecular weight. Isolation, purification and characterization of the amplified Akt kinase DNA can be performed by excising the fragment from the gel or eluting the fragment (for example, reference to Lawn et al., Nucleic Acids Research, 1981. 2: ρ·6103; Goeddel et al. Human, Nucleic Acids Research, 1980·8: ρ·4057); The amplified product is transferred into a suitable vector such as the pCRII vector (jnvitr〇gen); 2〇 will be transferred after transplantation The insert is sequenced; and the DNA sequence is compared to the known sequence of the LIM kinase. The relative amount of LIM kinase mRNA and cDNA was then determined. In one embodiment, real-time PCR is used to determine the degree of transcription of Akt nucleotides. The determination of transcriptional activity also includes assays based on available mRNA transcripts. Real-time PCR programs and other PCR programs use a variety of PCR products for detection chemistry, including binding of DNA-binding fluorophores, 5, endonucleases, adjacent linings and hairpin probes, and self-fluorescent amplification of genes . These chemical and real-time PCRs are generally discussed in Mackay et al., Nuclear 5-Acid Studies '2002·30(6): p.1292-1305; Walker, J Biochem Mol Maternal Science' 2001. 15(3): ρ. 121_127; Lewis et al, J Pathol, 2001. 195: p. 66-71. In another embodiment, the disordered expression of Akt can be identified by the following method: by allowing a nucleotide sequence isolated from a biological sample to have a map selected from 10th 6a-c, 7a-d, or 8a-c The oligonucleotide of the oligonucleoside gman sequence or the fragment thereof is contacted with a oligonucleotide probe of one of the sequences complementary thereto; and then the sequence is hybridized to the sequence via the probe and the result is compared to a normal sample. Hybridization of the probe to the biological sample can be detected by labeling the probe with any one of the detectable agents. The probe may, for example, be a radioisotope or be labeled with a biotin, a fluorescent dye, an electron dense reagent, an enzyme, a hapten or a protein which can be obtained. The detectable label can be assayed by any desired means including spectroscopic means, photochemical means, biochemical means, immunochemical means, radioisotope means, or chemical means. Probes can also be used, such as oligonucleotide restriction techniques, dot-point assays, reverse dot assays, 20-line probe assays, and 5' phosphatase assays. Alternatively, the probes can be detected using any of the commonly used DNA array technologies, including giant array technology, microarray technology, and DNA microchip technology. Oligonucleotide probes typically comprise about 14, 15, 16, 18, 20, 25 or 28 nucleotides that hybridize to nucleotides selected from the 6a-c, 7a-d and 8a-c maps or fragments thereof. Probes generally having a length of 67 200835507 greater than about 25 or 28 nucleotides are not preferred. Oligonucleotide probes are designed to recognize Akt nucleotide sequences. Kinase assay

Akt激酶之活性可使用技藝界已知之任一種適當激酶 5 檢定分析測定。例如但非限制性，可使用Hogg等人(致癌基因，1994· 9: p.98-96)，Mills等人(J Biol Chem，1992· 267: p.16000-006)及 Tomizawa 等人（FEBS Lett，2001，492: ρ·221·7)，Schmandt等人(J Immunol，1994. 152: ρ·96·105)所述方法。其它絲胺酸、蘇胺酸及酪胺酸激酶檢定分析述於 10 Ausubel等人（分子生物學之短協定方法，1999年，單元 17.6)。The activity of Akt kinase can be determined using any of the appropriate kinase 5 assay assays known to the art. For example and without limitation, Hogg et al. (Oncogene, 1994. 9: p. 98-96), Mills et al. (J Biol Chem, 1992. 267: p. 16000-006) and Tomizawa et al. (FEBS) can be used. Lett, 2001, 492: ρ·221·7), Schmandt et al. (J Immunol, 1994. 152: ρ·96·105). Additional assays for serine, threonine and tyrosine kinase assays are described in 10 Ausubel et al. (Short Protocol Methodology in Molecular Biology, 1999, unit 17.6).

Akt激酶檢定分析通常使用Akt多肽、經標記之施體酶基質、及5：體酶基質，該受體酶基質為Akt之特異性或非特 15 20 異性。於此種檢定分析中，Akt將經標記部分由施體酶基質轉移至受體酶基質，藉由施體酶基質轉移至受體酶基質之經過標記部分數量來測定_活性。鳩多肽可使用多種表現系統製造，可純化自細胞，可呈經裂解或未經裂解之組融合蛋白質形式及/或可有非·㈣肽序列例如脱標或_糖《於其N端或C端。若癌性細胞系用作為欲定分析之施的來源，則Akt活性可於癌性細齡中檢定析。適當Akt檢定分析狀施_基f &㈣讀Ak@Akt kinase assays typically employ an Akt polypeptide, a labeled donor enzyme substrate, and a 5:some enzyme matrix that is specific for Akt or non-specific. In this assay, Akt transfers the labeled moiety from the donor enzyme substrate to the acceptor substrate, and the activity is determined by the amount of labeled portion of the donor enzyme substrate transferred to the acceptor substrate. Purine polypeptides can be produced using a variety of expression systems, can be purified from cells, can be in the form of lysed or uncleaved fusion proteins, and/or can have non-(tetra)peptide sequences such as de-labeled or _ sugars at their N-terminus or C end. If a cancerous cell line is used as a source of the desired assay, Akt activity can be assayed for cancerous age. Appropriate Akt assay analysis _ base f & (four) read Ak@

酸化為敏感之任何分子，例如包括經T.標記之ATPAA 類似物，其中該標記為33p、/ b或任何其它放射性同素或適當螢光標記。Akt蚊分_之適t接受者峰基質 68 200835507 括對於藉Akt碎酸化為敏感之任—種乡肽或其它分子。接受者酶基貝係何生自活體内Akt標乾片段。接受者酶基質片段之長度可為8至5〇胺基酸，通常為聰如胺基酸，特別長約 10 12 15、丨8、20及25胺基酸。其它接受者酶基質可於 5實驗上使用—組不同的多肽或其它分子測定。TTK之接受者酶基質標乾可於一旦進行反應後純化自其它反應成分。此項純化通常係經由分子交互作用進行，此處接受者酶基質經過生物素化，且透過其與鏈絲菌抗生物素之交互作用來、屯化，或可取传特定抗體，該抗體可特異性識別接受者 1〇酶基質。反應係於多種條件下進行，諸如於固體撑體上，於謂、於溶液或於活細胞内進行。檢測方法之選擇係依據用於施體分子之標記類型決定，使用方法例如包括藉自動放射性攝影術、閃爍計數、掃描、或榮光攝影術測定所結合的放射性或螢光。 15 6·治療方法此處提供之化合物及藥學組成物可用於治療包括腫瘤症及其它與異常細胞增生相關聯之病症之情況。於 -個實施财’本發明化合物可用於治療_、肉瘤、淋巴瘤、白血病、及/或骨髓瘤。於本發明之其它實施例中， 20 此處揭示之化合物可用於治療實體腫瘤。本發明化合物可用於治療癌症，諸如官或組織之癌症攝護腺、肺、尿運、膀胱、非何杰金氏淋巴瘤、黑素瘤、腎、腎上腺、膜因頭、甲狀腺、胃、腦、多發性骨趟瘤、食道、肝、 69 200835507 肝内膽管、子宮頸、喉頭、急性骨髓性白血病、慢性骨髓性白血病、軟組織諸如心臟、何杰金氏淋巴瘤、睪丸、小腸、慢性骨髓性白血病、急性淋巴性白血病、肛門、肛管、肛門直腸、甲狀腺、陰門、膽囊、胸膜、眼、鼻、鼻腔、 5 中耳、鼻咽、輸尿管、腹膜、胃系膜、腸系膜、及胃腸道、高階神經膠瘤、神經膠母細胞瘤、大腸、直腸、胰、胃癌、肝細胞癌；頭頸癌、癌瘤；腎細胞癌；腎癌；肉瘤；血管内皮瘤；淋巴瘤；白血病、簟樣肉芽腫。於額外實施例中，本發明化合物可用於治療皮膚病包括但非限於惡性病血管 10 瘤、血管内皮瘤、基底細胞癌、鱗狀細胞癌、惡性黑素瘤、及卡波西氏肉瘤；及非惡性病或非惡性病症諸如乾癖、淋巴血管新生、兒童血管瘤、史土吉·偉博氏（Sturge-Weber) 症候群、尋常疣、神經纖維瘤、結節性硬化症、產膿性肉芽腫、隱性營養不良性大泡性表皮鬆解症、靜脈潰瘍、痤 15 瘡、酒潰鼻、濕療、傳染性軟；庞、脂漏性角化症、及光線性角化症。包括本發明化合物之組成物可用於由癌症的發現至惡化階段中之任何階段治療此等癌症及其它癌症。此外，包括本發明化合物之組成物可用於治療原發性癌症及轉移癌 20 症。於本發明之其它實施例中，此處所述化合物可用於治療癌症，包括但非限於下表2所列舉之癌症。 70 200835507Any molecule that is acidified to be sensitive, for example, includes a T.-tagged ATPAA analog, wherein the label is 33p, /b or any other radioactive element or a suitable fluorescent label. Akt Mosquito _ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ ̄ The recipient enzymes are derived from the Akt standard dry fragment in vivo. The acceptor enzyme substrate fragment can be from 8 to 5 amino acids in length, typically a succinct amino acid, particularly about 10 12 15 , 丨 8, 20 and 25 amino acids. Other recipient enzyme matrices can be assayed using 5 different sets of polypeptides or other molecules. The TTK acceptor enzyme substrate can be purified from other reaction components once the reaction is carried out. This purification is usually carried out via molecular interaction, where the recipient enzyme substrate is biotinylated and, through its interaction with streptavidin, sputum, or specific antibodies, the antibody can be specific The sexual recognition recipient 1 enzyme substrate. The reaction is carried out under a variety of conditions, such as on a solid support, in a solution, in solution or in living cells. The choice of detection method is determined by the type of label used for the donor molecule, and the method of use includes, for example, automated radiography, scintillation counting, scanning, or glory to determine the bound radioactivity or fluorescence. 15 6. Methods of Treatment The compounds and pharmaceutical compositions provided herein are useful in the treatment of conditions including oncology and other conditions associated with abnormal cell proliferation. The compounds of the invention may be used in the treatment of _, sarcoma, lymphoma, leukemia, and/or myeloma. In other embodiments of the invention, 20 the compounds disclosed herein are useful for treating solid tumors. The compounds of the present invention are useful for the treatment of cancer, such as cancer or prostate of the official or tissue, lung, urinary bladder, bladder, non-Hodgkin's lymphoma, melanoma, kidney, adrenal gland, membrane head, thyroid, stomach, brain Multiple osteoarthritis, esophagus, liver, 69 200835507 Intrahepatic bile duct, cervix, larynx, acute myeloid leukemia, chronic myelogenous leukemia, soft tissue such as heart, Hodgkin's lymphoma, testicular, small intestine, chronic myeloid Leukemia, acute lymphocytic leukemia, anal, anal canal, anorectal, thyroid, vulva, gallbladder, pleura, eye, nose, nasal cavity, 5 middle ear, nasopharynx, ureter, peritoneum, mesentery, mesentery, and gastrointestinal tract, High-grade neuroglioma, glioblastoma, large intestine, rectum, pancreas, gastric cancer, hepatocellular carcinoma; head and neck cancer, carcinoma; renal cell carcinoma; kidney cancer; sarcoma; hemangioendothelioma; lymphoma; leukemia, sputum-like granulation swollen. In additional embodiments, the compounds of the invention are useful in the treatment of dermatological conditions including, but not limited to, malignant disease vascular 10 tumors, hemangioendothelioma, basal cell carcinoma, squamous cell carcinoma, malignant melanoma, and Kaposi's sarcoma; Non-malignant or non-malignant conditions such as cognac, lymphangiogenesis, hemangioma in children, Sturge-Weber syndrome, vulgaris, neurofibromatosis, tuberous sclerosis, pyogenic granuloma , recessive dystrophic macrofocal epidermolysis, venous ulcers, sputum 15 sores, alcohol ulcers, wet treatment, infectious soft; Pang, lipid leakage keratosis, and linear keratosis. Compositions comprising a compound of the invention are useful for treating such cancers and other cancers at any stage from the discovery of the cancer to the stage of deterioration. Further, the composition comprising the compound of the present invention can be used for the treatment of primary cancer and metastatic cancer. In other embodiments of the invention, the compounds described herein are useful in the treatment of cancer, including, but not limited to, the cancers listed in Table 2 below. 70 200835507

急性淋巴母細胞性白血病，兒童急性骨髓性白血病，成人急性骨髓性白血病，兒童腎上腺皮質癌腎上腺皮質癌，兒童愛滋病相關癌症愛滋病相關淋巴瘤肛門癌星狀細胞瘤’兒童小腦星狀細胞瘤，兒童大腦 □基底細胞癌膽管癌，肝外膽癌膽癌，兒童骨癌骨肉瘤/惡性纖維性組織細胞瘤腦幹神經膠瘤，兒童腦瘤’成人腦瘤，腦幹神經膠瘤，兒童腦瘤，小腦星狀細胞瘤，兒童腦瘤，大腦星狀細胞瘤/惡性神經膠瘤，兒童腦瘤，室管膜瘤，兒童腦瘤，髓母細胞瘤，兒童腦瘤，小腦幕上未分化之神經外胚層瘤，兒童腦瘤，視覺徑路及下視丘神經膠瘤，兒童腦瘤，兒童乳癌乳癌，兒童乳癌，男性支氣管腺瘤/類癌，兒童肝細胞(肝癌），成人(原發性）肝細胞(肝癌），兒童(原發性）何杰金氏淋巴瘤，成人何杰金氏淋巴瘤，兒童何杰金氏淋巴瘤，懷孕期下咽癌下視丘及視覺徑路神經膠瘤，兒童 □眼内黑素瘤胰小島細胞癌(内分泌胰臟） □卡波西氏肉瘤腎(腎細胞)癌腎癌，兒童 □喉癌喉癌，兒童白血病，急性淋巴母細胞性，成人白血病’急性淋巴母細胞性，兒童白血病，急性骨髓性，成人白血病，急性骨髓性，兒童白血病，慢性淋巴細胞性白血病，慢性骨髓性白血病，B細胞唇及口腔癌肝癌，成人(原發性）肝癌，兒童(原發性）肝癌，非小細胞肝癌，小細胞淋巴瘤，愛滋病相關淋巴瘤，柏吉斯氏淋巴瘤淋巴瘤，皮膚T細胞，參考蕈狀肉芽腫及謝薩瑞(Sezary)症候群淋巴瘤，何杰金氏，成人淋巴瘤，何杰金氏，兒童淋巴瘤，何杰金氏，懷孕期 71 200835507 柏吉斯氏(Burkitt’s)淋巴瘤 □類癌瘤，兒童類癌瘤，胃腸道未知原發性中樞神經系統癌淋巴瘤，原發性小腦，星狀細胞瘤，兒童大腦，星狀細胞瘤/惡性神經膠瘤，兒童子宮頸癌兒童期癌症慢性淋巴細胞性白血病慢性骨髓性白血病慢性骨髓增生病症大腸癌大腸直腸癌，兒童皮膚T細胞淋巴瘤，參考輩狀肉芽腫及謝薩瑞症候群 □子宮内膜癌室管膜瘤，兒童食道癌食道癌，兒童歐文氏(Ewing’s)腫瘤家族顱外胚細胞腫瘤，兒童性腺外胚細胞腫瘤肝外膽管癌眼癌，眼内黑素瘤眼癌，視網膜母細胞瘤 □膽囊癌胃癌胃癌，兒童胃腸道類癌腫瘤胚細胞腫瘤，顱外，兒童胚細胞腫瘤，性腺外胚細胞腫瘤，印巢妊娠滋養層腫瘤神經膠瘤，成人神經膠音腦鈐淋巴瘤，非何杰金氏，成人淋巴瘤，非何杰金氏，兒童淋巴瘤，非何杰金氏，懷孕期淋巴瘤，原發性中樞神經系統 □巨球蛋白血症，沃登史壯氏 (Waldenstrom’s)惡性纖維性骨組織細胞瘤/骨肉瘤髓母細胞瘤，兒童黑素瘤黑素瘤，眼内(眼）默可(Merkel)細胞癌間皮瘤，成人惡性間皮瘤，兒童帶有隱藏原發性轉移性鱗狀細胞頸癌多發性内分泌腫瘤生成症狀群，兒童多發性骨髓瘤/漿細胞瘤蕈狀肉芽腫骨髓發育不良症候群骨髓發育不良/骨髓增生病骨髓性白血病，慢性骨鰱性白血病，成人急性骨髓性白血病，兒童急性骨髓瘤，多發性骨髓增生病症，慢性 □鼻腔及副鼻竇癌鼻咽癌鼻咽癌，兒童神經母細胞瘤非何杰金氏淋巴瘤，成人非何杰金氏淋巴瘤，兒童非何杰金氏淋巴瘤，懷孕期非小細胞肺癌 □ 口癌，兒童口腔癌，唇及口咽癌骨肉瘤/惡性纖維性骨組織細胞瘤卵巢癌，兒童 72 200835507 神經膠瘤，兒童大腦卵巢上皮癌星狀細胞瘤印巢胚細胞腫瘤神經膠瘤，兒童視覺徑路及下視丘卵巢低惡性潛在腫瘤 □皮膚癌(黑素瘤） □騰癌皮膚癌，默可細胞胰癌，兒童小細胞肺癌騰癌，騰島細胞小腸癌副鼻竇及鼻腔癌軟組織肉瘤，成人副甲狀腺癌軟組織肉瘤，兒童陰莖癌鱗狀細胞癌，參考皮膚癌(非黑素瘤）嗜鉻細胞瘤有隱藏原發性之鱗狀細胞頸癌，轉松果體母細胞瘤及小腦幕未分化神移經外胚層腫瘤，兒童胃癌腦垂腺腫瘤胃癌，兒童漿細胞瘤/多發性骨髓瘤小腦幕上未分化神經外胚層腫瘤，胸膜肺母細胞瘤兒童懷孕及乳癌 □T細胞淋巴瘤，皮膚，參考簟狀肉懷孕及何杰金氏淋巴瘤芽腫及謝薩瑞症候群懷孕及非何杰金氏淋巴瘤睪丸癌原發性中樞神經系統淋巴瘤胸腺瘤，兒童攝護腺癌胸腺瘤及胸腺癌 □直腸癌甲狀腺癌腎細胞(腎)癌甲狀腺癌，兒童腎細胞(腎)癌，兒童腎盂與輸尿管之過渡細胞癌腎盂及輸尿管，過渡細胞癌滋養層腫瘤，姓娠視網膜母細胞瘤 □未知原發位置，癌，成人橫紋肌肉瘤，兒童未知原發位置，癌，兒童 □唾液腺癌兒童之非尋常癌唾液腺癌，兒童輸尿管及腎盂，過渡細胞癌肉瘤，歐文氏腫瘤家族尿道癌肉瘤，卡波西氏子宮癌，子宮内膜肉瘤，軟組織，成人子宮肉瘤肉瘤，軟組織，兒童 □***癌肉瘤，子宮視覺徑路及下視丘神經膠瘤，兒童謝薩瑞症候群陰門癌皮膚癌(非黑素瘤） □沃登史壯氏巨球蛋白血症威姆氏 (Wilms’)腫瘤皮膚癌，兒童 73 200835507 於本發明之其它實施例中，此處揭示化合物可用於治療血管新生相關之疾病。抗血管新生小分子包括沙利竇邁(thalidomide)，其部分經由抑制NFkB發揮作用；2-曱氧基***影響微管的形成 5 及缺氧誘導因子（HIFla)之活化；環氧合酶2(COX2)抑制劑；及低劑量習知化學治療劑包括環磷醯胺、紫杉烷類 (taxanes)及長春花生物驗（文克里斯丁（vincristine)、文布拉斯丁（vinblastine) (D’Amato R.J·等人，Proc.Natl. Acad· Sci. U.S.A·，1994. 91: p.3964-3968; D，Amato R.J.等人，Proc.Natl. 10 Acad· Sci· U.S.A·，1994· 91: ρ·4082-4085)。此外，某些酪胺酸激酶抑制劑經由減少腫瘤及基質細胞製造VEGF及其它前血管新生因子的產量來間接降低血管新生。此等藥物包括贺癌平(Herceptin)、伊馬堤尼(imatinib)(葛里費（Glivec)) 及艾瑞莎（Iressa) (Bergers G.等人，J Clin Invest，2003.111: 15 P.1287-1295; Ciardiello F.等人，臨床癌症研究，2〇〇ι·7: p.1459-1465; Plum S.M·等人，臨床癌症研究，2〇〇39: ρ·4619_4626)。晚近，血管新生抑制劑已經由動物研究模型移至人類病人的研究。血管新生抑制劑代表多種不同癌症的有展望 20之治療方法。晚近阿法斯丁（Avastin) —種對血管内皮生長因子(VEGF)有高度親和力之抗體，阿法斯丁顯示用於惡化的腎細胞癌作為單一藥劑可延長壽命，用於惡化的大腸癌結合化學治療可延長壽命(Yang j.C·等人，新英格蘭期刊Acute lymphoblastic leukemia, childhood acute myelogenous leukemia, adult acute myelogenous leukemia, adrenocortical carcinoma of the adrenaline, AIDS-related cancer, AIDS-associated lymphoma, anal cancer, astrocytoma, children's cerebellar astrocytoma, children Brain basal cell carcinoma cholangiocarcinoma, extrahepatic cholangiocarcinoma, childhood bone cancer osteosarcoma / malignant fibrous histiocytoma brain stem neuroglioma, childhood brain tumor ' adult brain tumor, brain stem glioma, childhood brain tumor , cerebellar stellate cell tumor, childhood brain tumor, brain astrocytoma / malignant glioma, childhood brain tumor, ependymoma, childhood brain tumor, medulloblastoma, childhood brain tumor, undifferentiated on the cerebellum Neuroectodermal tumor, childhood brain tumor, visual path and hypothalamic glioma, childhood brain tumor, childhood breast cancer, childhood breast cancer, male bronchial adenoma / carcinoid, childhood liver cell (liver cancer), adult (primary Sex) Hepatocytes (liver cancer), children (primary) He Jiejin's lymphoma, adult Hodgkin's lymphoma, child He Jiejin's lymph Tumor, hypopharyngeal hypothalamic and visual pathway neurogyngoma during pregnancy, children with intraocular melanoma and pancreatic islet cell carcinoma (endocrine pancreas) □ Kaposi's sarcoma kidney (kidney cell) cancer, renal cancer, children Throat laryngeal cancer, childhood leukemia, acute lymphoblastic, adult leukemia 'acute lymphoblastic, childhood leukemia, acute myeloid, adult leukemia, acute myeloid, childhood leukemia, chronic lymphocytic leukemia, chronic myelos Leukemia, B cell lip and oral cancer liver cancer, adult (primary) liver cancer, children (primary) liver cancer, non-small cell liver cancer, small cell lymphoma, AIDS-associated lymphoma, baijis lymphoma, Skin T cells, reference to sputum granuloma and Sezary syndrome lymphoma, He Jiejin, adult lymphoma, He Jiejin, childhood lymphoma, He Jiejin, pregnancy 71 200835507 Peregis Burkitt's lymphoma 类 carcinoid tumor, childhood carcinoid tumor, gastrointestinal tract unknown primary central nervous system cancer lymphoma, primary cerebellum, stellate cell tumor Children's brain, astrocytoma/malignant glioma, childhood cervical cancer, childhood cancer, chronic lymphocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorders, colorectal cancer, colorectal cancer, childhood cutaneous T-cell lymphoma, reference generation granulation Swollen and Xie Saray syndrome □ endometrial cancer ependymoma, esophageal cancer esophageal cancer in children, Ewing's tumor family cranial blast cell tumor, childhood gonadal blast cell tumor extrahepatic cholangiocarcinoma eye cancer, eye Endometrial cancer, retinoblastoma, gallbladder carcinoma, gastric cancer, gastrointestinal carcinoid tumor, blast, tumor, extracranial, childhood blast, gonadal blast, tumor, gestational trophoblastic neoplasm , adult nerve gel cerebral palsy lymphoma, non-Hodgkin's, adult lymphoma, non-Hodgkin's, childhood lymphoma, non-Hodgkin's, lymphoma of pregnancy, primary central nervous system □ giant ball Proteinemia, Waldenstrom's malignant fibrous bone histiocytoma / osteosarcoma medulloblastoma, child black Tumor melanoma, intraocular (eye) Merkel cell carcinoma mesothelioma, adult malignant mesothelioma, children with multiple metastatic squamous cell carcinoma, multiple endocrine neoplasia syndrome, children Multiple myeloma/plasma tumor verrucous granuloma myelodysplastic syndrome bone marrow dysplasia/myeloproliferative myeloid leukemia, chronic osteomyelitis, acute myeloid leukemia in adults, acute myeloma in children, multiple myeloproliferative disorders, Chronic nasopharyngeal and paranasal sinus cancer nasopharyngeal carcinoma nasopharyngeal carcinoma, childhood neuroblastoma non-Hodgkin's lymphoma, adult non-Hodgkin's lymphoma, non-Hodgkin's lymphoma in children, non-small cells during pregnancy Lung cancer □ Oral cancer, oral cancer in children, lip and oropharyngeal cancer Osteosarcoma/malignant fibrous bone histiocytoma ovarian cancer, child 72 200835507 Neuroglioma, children's cerebral ovarian epithelial carcinoma, astrocytoma, blastocyst, tumor glial Tumor, children's visual path and hypothalamic ovarian low malignant potential tumor □ skin cancer (melanoma) □ TB cancer skin cancer, Merck cells Cancer, small cell lung cancer in children, Tengdao cell small intestine cancer, paranasal sinus and nasal cancer soft tissue sarcoma, adult parathyroid cancer soft tissue sarcoma, childhood penile cancer squamous cell carcinoma, reference skin cancer (non-melanoma) pheochromocytoma There are hidden primary squamous cell carcinoma, transgenic pineal blastoma and cerebellar undifferentiated God through ectodermal tumor, childhood gastric cancer, sinus adenocarcinoma, gastric cancer, childhood plasmacytoma / multiple myeloma cerebellum Upper undifferentiated neuroectodermal tumor, pleural pulmonary blastoma, childhood pregnancy and breast cancer □ T-cell lymphoma, skin, reference to sputum meat pregnancy and Hodgkin's lymphoma bud and Xie Saray syndrome pregnancy and non-Ho Jerkin Lymphoma, squamous cell carcinoma, primary central nervous system lymphoma, thymoma, childhood prostate cancer, thymoma, thymic carcinoma, rectal cancer, thyroid cancer, renal cell (kidney) cancer, thyroid cancer, childhood renal cell (kidney) cancer, childhood pyelone Transitional cell carcinoma with ureter, renal pelvis and ureter, transitional cell carcinoma trophoblastic tumor, surname retinoblastoma □ unknown original Location, cancer, adult rhabdomyosarcoma, unknown origin of children, cancer, children, salivary adenocarcinoma, abnormal cancer, salivary gland cancer, ureter and renal pelvis, transitional cell carcinoma, Owen's tumor, urethral carcinosarcoma, Kaposi Uterine cancer, endometrial sarcoma, soft tissue, adult uterine sarcoma sarcoma, soft tissue, children □ vaginal carcinosarcoma, uterine visual pathway and hypothalamic glioma, children with Xie Saray syndrome, gland cancer, skin cancer (non-melanoma) □ Warden's giant globulinemia Wilms' tumor skin cancer, child 73 200835507 In other embodiments of the invention, the compounds disclosed herein are useful for treating diseases associated with angiogenesis. Anti-angiogenic small molecules include thalidomide, which partially acts by inhibiting NFkB; 2-methoxy estradiol affects microtubule formation 5 and activation of hypoxia-inducible factor (HIFla); Enzyme 2 (COX2) inhibitors; and low doses of conventional chemotherapeutic agents include cyclophosphamide, taxanes, and periwinkle bioassay (vincristine, vinblastine) (D'Amato RJ et al., Proc. Natl. Acad. Sci. USA, 1994. 91: p. 3964-3968; D, Amato RJ et al., Proc. Natl. 10 Acad·Sci· USA·, 1994· 91: ρ·4082-4085). In addition, certain tyrosine kinase inhibitors indirectly reduce angiogenesis by reducing the production of VEGF and other pro-angiogenic factors by tumors and stromal cells. (Herceptin), Imatinib (Glivec) and Iressa (Bergers G. et al., J Clin Invest, 2003.111: 15 P.1287-1295; Ciardiello F. et al. Human, Clinical Cancer Research, 2〇〇ι·7: p.1459-1465; Plum SM· et al, Clinical Cancer Research, 2〇〇39: ρ· 4619_4626). Recently, angiogenesis inhibitors have been moved from animal research models to human patients. Angiogenesis inhibitors represent a variety of different cancers with a prospective 20 treatment. Late Avastin (Avastin) Growth factor (VEGF) has a high affinity antibody, and Avastin shows that renal cell carcinoma for worsening as a single agent can prolong lifespan, and can be used for prolonged colorectal cancer combined with chemotherapy to prolong life (Yang jC· et al., New English Journal

Med，2003· 349: p.427-434; Kabbinavar F.等人，j Clin One 74 200835507 2003· 21: p.60-65)。血官新生相關疾病包括但非限於發炎病、自體免疫病及傳杂病；血管新生依賴型癌症包括例如實體腫瘤、血液系統腫瘤諸如白血病及腫瘤轉移；良性腫瘤諸如血管瘤、 5聽神經瘤、神經纖維瘤、氣管瘤、及產膿性肉芽腫；類風濕性關節炎；乾癬；濕疹；眼血管新生病例如糖尿病性視網膜病變、早產視網膜病變、黃斑部退化、角膜移植排斥、新生血管性青光眼、晶狀體後纖維增生、虹膜紅變症；奥斯路-¼博(0sler_Webber)症候群；心肌血管新生；斑塊新生 1〇血官化；毛細血管擴張；嗜血性關節；血管纖維瘤；及傷口肉芽腫化。此外，本發明組成物可用於治療下列疾病諸如但非限於腸沾黏、動脈粥狀硬化、硬皮病、；庑及肥厚性瘢痕（亦即瘢瘤）。本發明組成物也可用於治療由於病理後果出現血管新生之病症，諸如貓抓熱（小型五日羅謝爾菌 15 (Rochele minalia quintosa))、潰瘍（胃幽門螺旋桿菌 (Helobacter pylori))、結核病及狼瘡。 6·1抗藥性腫瘤或癌症之治療本發明提供可用於治療抗藥性癌症之化合物，包括癌症實施例及曲西立濱化合物及/或爾洛堤尼系列化合物例 2〇如傑費堤尼、爾洛堤尼或其鹽。多重抗藥性(MDR)出現於人類癌症，可能是化學治療成功的主要障礙。多重抗藥性是一種腫瘤細胞於試管内暴露於一種細胞毒劑發展出對結構上相關及功能上相關之一定範圍之化合物具有交叉抗藥性的現象。此外多重抗藥性 75 200835507 也可能特有地發生 , 《系些癌症而未先前暴露於化學治療剤。如此，於一個實 .M列中，本發明提供藉投予TCN、TCN-P 或相關化合物及爾、、夂 α 口 4 ^ 〜疋尼系列化合物例如傑費堤尼、爾洛土疋尼或其鹽治療s μ〜^藥性癌症例如多重抗藥性癌症之病人之方法。於若干徐汽施例中，TCN、TCN-P及相關化合物及爾洛堤尼系列化八 α物例如傑費堤尼、爾洛堤尼或其鹽可用於治療單獨對紫私、拉帕黴素、塔莫西芬、西鉑汀及/ 或傑:是尼(艾瑞莎)有抗藥性之癌症。於一個實施例中，TCN、TCN-P或相關化合物及爾洛 10 %尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可用於治療抗藥性結腸癌、骨癌、腎癌、腎上腺癌、胰癌、肝癌及/ 或任何其它技藝界已知或此處所述之癌症。 6·2·組合治療於一個實施例中，曲西立濱化合物及爾洛堤尼系列化 15合物例如傑費堤尼、爾洛堤尼或其鹽玎連同其它細胞毒劑投予。於另一個實施例中，曲西立濱化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽及其組成物當用於治療實體腫瘤時，可使用輻射投予。於本發明之另一個實施例中，此處揭示之曲西立濱化 2〇合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽及其組成物可組合至少一種額外化學治療劑投予。額外藥劑可與此處揭示之化合物組合投予或交替投予。藥物可構成同種組成物之一部分，或可於同時或不同時呈分開組成物投予。 76 200835507 於一個實施例中，如此處揭示之曲西立濱化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可組合抗血管新生劑來提升其效果，或組合其它抗血管新生劑且連同其它細胞毒劑一起投予。於另一個實施例中，曲西立 5 濱化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽及組成物當用於治療實體腫瘤時可與選自於但非限於下列藥劑投予·· IL-12、維生素A酸類、干擾素類、血管抑制素、内皮抑制素、沙利竇邁、血栓素 (thrombospondin)-l、血栓素-2、卡普脫普利（captopryl)、抗 10 腫瘤劑諸如α干擾素、COMP(環磷醯胺、文克里斯丁、甲胺嗓呤及普尼松（prednisone))、伊妥普賽（etoposide)、 mBACOD(甲胺嗓吟、布利歐黴素(bleomycin)、朵索魯冰心 (doxorubicin)、環磷醯胺、文克里斯丁及德沙美沙松 (dexamethasone))、PRO-MACE/MOPP(普尼松、甲胺喋呤(含 15 路可文(leucovin)救援）、朵索魯冰心、環麟醯胺、伊妥普賽 /美可羅薩敏(mechlorethamine)、文克里斯丁、普尼松及普卡巴辛(procarbazine))、文克里斯丁、文布拉斯丁、血管抑制素、TNP-470、多硫酸戊聚糖、血小板因子4、血管抑制素、LM-609、SU-101、CM-101、泰嘉蘭（Techgalan)、沙利 20 竇邁、SP-PG及放射線。於額外實施例中，此處揭示之化合物及組成物可與下列藥物組合投予或交替投予：例如具有抗有絲***效果之藥物諸如標把細胞骨胳元素，包括鬼白毒或長春花生物鹼(文克里斯丁、文布拉斯丁）；抗代謝藥物 (諸如5-氟尿喊σ定、赛它拉冰（cytarabine)、傑西塔賓 77 200835507 (gemcitabine)、嘌呤類似物諸如戊抑制素(pent〇statin)、曱胺喋呤）；烷化劑或氮芥子氣(諸如亞硝基脲、環磷醯胺、或伊福法邁（ifosphamide));靶定於DNA之藥物諸如蔥環素 (antracycline)藥物亞利亞黴素(adriamycin)、朵索魯冰心、 5 法母魯冰心(pharmorubicin)或伊皮魯冰心（epirubicin);乾定於拓樸異構酶之藥物諸如伊妥普賽；激素及激素激動劑或激素拮抗劑諸如***類、抗***類（塔莫西芬及相關化合物）及雄激素類、福塔邁（flutamide)、路普瑞林 (leuProrelln)、葛瑟瑞林(goserelin)、賽普左（cyprotrone)、或 10 歐克萃泰（octreotide);靶定於腫瘤細胞之信號轉導之藥物，包括抗體衍生物諸如贺癌平；烷化藥物諸如鉑藥物（西鉑汀、卡朋伯丁（carbonplatin)、歐薩里伯丁（oxaiiplatin)、帕拉伯丁(paraplatin))或亞硝基脲類；可能影響腫瘤轉移之藥物，諸如基質金屬蛋白酶抑制劑；基因治療及反訊息劑； 15抗體治療；其中海洋來源之生物活性化合物值得注意者為戴尼斯（didemnins)諸如阿普里丁（aplidine);類固醇類似物特別為德沙美沙松；抗炎藥物包括非類固醇藥劑(諸如乙醯胺酚(acetaminophen)或伊布普芬(ibUprofen))或類固醇及其何生物特別為德沙美沙松；止吐藥包括5HT-3抑制劑（諸如 2〇葛蘭米色壯(gramisetron)或盎達色壯(ondasetron))及類固醇及其衍生物特別為德沙美沙松。又有其它實施例中，化合物及組成物可與下表3所揭示之化學治療劑組合投予或六替投予。 /乂 78 200835507Med, 2003· 349: p. 427-434; Kabbinavar F. et al., j Clin One 74 200835507 2003· 21: p. 60-65). Related blood diseases include, but are not limited to, inflammatory diseases, autoimmune diseases, and transmission diseases; angiogenesis-dependent cancers include, for example, solid tumors, hematological tumors such as leukemia and tumor metastasis; benign tumors such as hemangiomas, 5 acoustic neuromas, Neurofibromatosis, tracheal tumor, and pyogenic granuloma; rheumatoid arthritis; cognac; eczema; ocular neovascularization such as diabetic retinopathy, recurrent retinopathy, macular degeneration, corneal transplant rejection, neovascularization Glaucoma, posterior lens hyperplasia, iris red syndrome; Oslo-1⁄4 Bo (0sler_Webber) syndrome; myocardial angiogenesis; plaque newborn 1 〇 blood stasis; telangiectasia; bloodthirsty joint; angiofibroma; Granuloma. Furthermore, the compositions of the present invention are useful in the treatment of diseases such as, but not limited to, intestinal adhesion, atherosclerosis, scleroderma, sputum and hypertrophic scars (i.e., tumors). The compositions of the invention may also be used to treat conditions of angiogenesis due to pathological consequences, such as cat scratching (Rochele minalia quintosa), ulcers (Helobacter pylori), tuberculosis And lupus. 6.1 Treatment of Drug-Resistant Tumors or Cancers The present invention provides compounds useful for the treatment of drug-resistant cancers, including cancer embodiments and triclinib compounds and/or erlotini series compounds, such as Gefitini, Erlotini or its salt. Multiple drug resistance (MDR) occurs in human cancer and may be a major obstacle to the success of chemotherapy. Multidrug resistance is a phenomenon in which a tumor cell is exposed to a cytotoxic agent in a test tube to develop a cross-resistance to a compound that is structurally and functionally related. In addition, multiple drug resistance 75 200835507 may also occur uniquely, "the cancer is not previously exposed to chemotherapy." Thus, in a real M column, the invention provides for the administration of TCN, TCN-P or related compounds, and the 4α 4 ^ 疋系列 series of compounds such as gefitini, erlotote or A method in which a salt thereof treats a patient with a drug-resistant cancer such as a multidrug resistant cancer. In several Xu steam applications, TCN, TCN-P and related compounds and erlotini series of eight alpha compounds such as gefitini, erlotini or its salts can be used to treat singly , Tamoxifen, West Platin and/or Jay: It is a cancer that is resistant to Nie (Aresa). In one embodiment, TCN, TCN-P or related compounds and Ello 10% series of compounds such as gefitinib, erlotinib or a salt thereof can be used for the treatment of drug resistant colon cancer, bone cancer, kidney cancer, adrenal gland Cancer, pancreatic cancer, liver cancer, and/or any other cancer known to the artisan or described herein. 6.2 Combination Therapy In one embodiment, the triclinide compound and the erlotinib-like compound such as gefitinib, erlotinib or its salt mash are administered along with other cytotoxic agents. In another embodiment, the triclinide compound and the erlotiniline compound, such as gefitinib, erlotinib or a salt thereof, and compositions thereof, when used to treat a solid tumor, can be administered using radiation. In another embodiment of the present invention, the triclinicin 2 conjugate disclosed herein and the erlotini series of compounds such as gefitini, erlotini or a salt thereof and combinations thereof may be combined at least An additional chemotherapeutic agent is administered. Additional agents can be administered or administered in combination with the compounds disclosed herein. The drug may form part of the same composition or may be administered as a separate composition at the same time or at different times. 76 200835507 In one embodiment, the triclinide compound and the erlotini series of compounds such as gefitinib, erlotinib or a salt thereof as disclosed herein may be combined with an anti-angiogenic agent to enhance the effect, or a combination thereof. Other anti-angiogenic agents are administered along with other cytotoxic agents. In another embodiment, the triclinide compound and the erlotini series of compounds such as gefitinib, erlotinib or a salt thereof and a composition thereof, when used in the treatment of a solid tumor, may be selected from the group consisting of Limited to the following pharmaceutical administrations · IL-12, retinoic acid, interferon, angiostatin, endostatin, salipirin, thromboxane-l, thromboxane-2, captopril (captopryl), anti-10 tumor agents such as alpha interferon, COMP (cyclophosphamide, bicinchine, methotrexate and prednisone), etoposide, mBACOD (methylamine)嗓吟, bleomycin, doxorubicin, cyclophosphamide, christaldine and dexamethasone, PRO-MACE/MOPP (Punisson, A Amine (including 15 leucovin rescue), Dorothy Bing, Halal, Mechlorethamine, Vincent, Prenisson and Puqaba Procarbazine, venscridin, vonbrastin, angiostatin, TNP-470, pentosan polysulfate, platelets Sub-4, angiostatin, LM-609, SU-101, CM-101, Taijia Lan (Techgalan), thalidomide sinus step 20, SP-PG and radiation. In additional embodiments, the compounds and compositions disclosed herein may be administered or administered in combination with the following agents: for example, drugs having anti-mitotic effects such as standard cell bone elements, including ghost white or vinca alkaloids. (Wen Christian, von Brastin); antimetabolites (such as 5-fluorourine sputum, cytarabine, Jessitabin 77 200835507 (gemcitabine), purine analogs such as pentatin (pent〇statin), amidoxime); alkylating agent or nitrogen mustard gas (such as nitrosourea, cyclophosphamide, or ifosphamide); drugs targeting DNA such as onionine ( Antracycline) drug adriamycin (adriamycin), dossolu ice heart, 5 pharmorubicin or epirubicin; a drug that is determined by topoisomerase such as itopride; Hormone and hormone agonists or hormone antagonists such as estrogens, antiestrogens (tamoxifen and related compounds) and androgens, flutamide, leuprorelln, gersui Forest (goserelin), cyprotron (cyprotron e), or 10 octreotide; drugs targeted for signal transduction of tumor cells, including antibody derivatives such as carbamazepine; alkylating drugs such as platinum drugs (xiplatin, carpenter ( Carbonplatin), oxaiiplatin, paraplatin or nitrosourea; drugs that may affect tumor metastasis, such as matrix metalloproteinase inhibitors; gene therapy and anti-message; 15 antibodies Treatment; wherein marine-derived bioactive compounds are notable for didemnins such as aplidine; steroid analogs are particularly desoxasone; anti-inflammatory drugs include non-steroidal agents (such as acetaminophen) Or ibuprol (ibUprofen) or steroids and their organisms especially for dexamethasone; antiemetics include 5HT-3 inhibitors (such as 2 gram gramisetron or ondasetron) And steroids and their derivatives are especially desoxasone. In still other embodiments, the compounds and compositions can be administered in combination or in combination with the chemotherapeutic agents disclosed in Table 3 below. /乂 78 200835507

表3 :化學治療劑 -13-順•維生素A酸 -2-胺基-6-綺基嗓σ令 -2-CdA -2-氯去氧腺苔 -5-氟尿嘴。定 -5-FU -6-TG -6-鳥嘌呤 -6-M基嘌呤 -6-MP -阿奎坦(Accutane) -放線黴素(Actinomycin)-D -亞利亞黴素(Adriamycin) -亞杜習(Adrucil) -亞葛林(Agrylin) -阿拉可(Ala-Cort) -亞德路金(Aldesleukin) -亞蘭土竹美(Alemtuzumab) -亞里堤諾(Alitretinoin) -阿卡班(Alkaban)-AQ _ 阿卡蘭(Alkeran) -全反式維生素A酸 -α干擾素 -亞特塔明(Altretamine) -阿米索泰林(Amethopterin) -阿米福斯丁(Amifostine) -阿米諾谷堤邁(Aminoglutethimide) -阿納葛萊(Anagrelide) -阿納左(Anandron) -阿納史左(Anastrozole) -***糖基胞苷 -Ara-C -阿拉尼斯普(Aranesp) -阿瑞戴(Aredia) -阿瑞米戴斯(Arimidex) -阿羅馬辛(Aromasin) -三氧化砷 -尼薩(Neosar) -努拉斯塔(Neulasta) _努美加(Neumega) -努普金(Neupogen) _尼蘭左(Nilandron) -尼魯塔邁(Nilutamide) -氮芥子氣 -諾法戴(Novaldex) -諾凡左(Novantrone) -歐萃泰(Octreotide) -乙酸歐卒泰 -歐可斯巴(Oncospar) •歐可文(Oncovin) -歐塔(Ontak) -歐薩(Onxal) -歐普維金(Oprevelkin) -歐拉普(Orapred) -歐拉松(Orasone) -歐薩里伯丁(Oxaliplatin) -太平洋紫杉醇(Paclitaxel) -帕里左奈(Pamidronate) -潘瑞丁(Panretin) -帕拉伯丁 (Paraplatin) -皮戴普(Pediapred) -PEG干擾素 -佩嘉史帕蓋(Pegaspargase) -沛福葛拉斯丁 (Pegfilgrastim) -PEG-INTRON -PEG-L-天冬酿胺酶 -苯基丙二酸芥子氣 -普拉堤諾(Platinol) -普拉堤諾-AQ -普尼松隆(Prednisolone) -普尼松(Prednisone) -普隆(Prelone) -普卡巴辛(Procarbazine) -PROCRIT 79 200835507 -天冬醯胺酶 -ATRA -阿法斯丁 (Avastin) -BCG -BCNU -貝法西組馬(Bevacizumab) -貝薩羅丁(Bexarotene) -拜卡路塔邁(Bicalutamide) -BiCNU -布諾山(Blenoxane) -布歐黴素(Bleomycin) -布特左米(Bortezomib) -布蘇坊(Busulfan) -布蘇費(Busulfex) -C225 -路可福瑞(Leucovorin)i丐 -坎帕斯(Campath) -坎妥薩(Camptosar) -喜樹鹼(Camptothecin)-11 -開普西塔冰(Capecitabine) -卡拉(Carac) -卡朋伯丁(Carboplatin) -卡幕斯丁 (Carmustine) -卡幕斯丁糊 -卡索戴(Casodex) -CCNU -CDDP -西努(CeeNU) -西魯比丁(Cembidine) •西土席馬(cetuximab) •可羅拉布席(Chlorambucil) -西鈾 '汀(Cisplatin) -水化葉酸因子(Citrovorum Factor) -可拉左冰(Cladribine) -可體松(Cortisone) -可美金(Cosmegen) -CPT-11 -環碟醯胺(Cyclophosphamide) -赛塔左恩(Cytadren)_ -普路金(Proleukin) -有卡幕斯丁(Carmustine)植體之普利菲普斯潘(Prolifeprospan) 20 -普瑞尼索(Purinethol) •拉羅西芬(Raloxifene) -魯馬萃(Rheumatrex) -瑞土山(Rituxan) -瑞土西馬(Rituximab) -瑞福隆(Roveron)-A(干擾素a -2a) -魯貝(Rubex) -鹽酸魯畢朵黴素(Rubidomycin) -山朵史塔丁 (Sandostatin) -山朵史塔丁 LAR -薩葛幕斯丁 (Sargramostim) -蘇路-可堤(Solu-Cortef) -蘇路-美左(S〇lu-Medrof) -STI-571 -史翠妥左辛(Streptozocin) -塔莫西芬(Tamoxifen) •塔可丁 (Targretin) -紫杉醇(Taxol) -紫杉帖(Taxotere) -堤莫達(Temodar) -堤莫左羅米(Temozolomide) -堤尼普赛(Teniposide) -TESPA -沙利竇邁(Thalidomide) -薩羅米(Thalomid) -席拉赛(TheraCys) -硫鳥嘌呤 -硫鳥嘌呤録； -硫磷醯胺(Thiophosphoamide) -硫普歹J (Thioplex) -硫泰帕(Thiotepa) -TICE -妥普薩(Toposar) -妥普堤肯(Topotecan) -妥瑞米芬(Toremifene) -崔茲竹美(Trastuzumab)_ 80 200835507 -赛塔拉賓(Cytarabine) -崔堤諾(Tretinoin) -赛塔拉賓微脂粒 -崔薩(Trexall) -賽妥色(Cytosar)-U _ 崔森諾(Trisenox) -賽妥山(Cytoxan) -TSPA -達卡巴辛(Dacarbazine) -VCR -達堤諾黴素(Dactinomycin) -維班(Velban) _達本普丁 (Darbepoetin) α -維卡德(Velcade) -道諾黴素(Daunomycin) -維普西(VePesid) -鹽酸道諾魯冰心(Daunorubicin) -維莎諾(Vesanoid) -道諾魯冰心微脂粒 -維杜爾(Viadur) -道諾索(DaunoXome) -硫酸文布拉斯丁 (Vinblastine) -德卡左(Decadron) -文卡薩(Vincasar)Pfs -戴它可泰(Delta-Cortef) -文克里斯丁 (Vincristine) -戴它松(Deltasone) -文諾瑞賓(Vinorelbine) -戴尼魯金(Denileukin)戴堤妥(diftitox) -酒石酸文諾瑞賓 -德普赛(DepoCyt) -VLB -德沙美沙松(Pexamethasone) -VP· 16 -乙酸德沙美沙松 -福蒙(Vumon) -磷酸德沙美沙松鈉 -贊羅達(Xeloda) -德沙松(Dexasone) •山諾薩(Zanosar) -戴拉左山(Dexrazoxane) -山法林(Zevalin) -DHAD -辛卡(Zinecard) -DIC -左拉戴(Zoladex) -戴德(Diodex) -左朵尼酸(Zoledronic acid) -朵西紫杉醇(Docetaxel) -左米塔(Zometa) -朵席(Doxil) -葛里戴(Gliadel)糊 -朵索魯冰心(Doxorubicin) -葛里費(Glivec) -朵索魯冰心微脂粒 -GM-CSF -左賽亞(proxia) •葛瑟瑞林(Goserelin) -DTIC •粒狀細胞-群落刺激因子 -DTIC-朵母(Dome) -粒狀細胞-巨噬細胞-群落刺激因子 -杜拉隆(Duralone) -哈羅泰斯丁 (Halotestin) -伊福戴(EfUdex) -賀癌平(Herceptin) -伊萊嘉^ligard) -賀薩左(Hexadrol) -艾倫斯(Ellence) -贺薩蘭(Hexalen) -艾羅薩丁 (Eloxatin) -六甲基蜜胺 -艾斯巴(Elspar) -HMM •安賽特(Emcyt) -海坎丁 (Hycamtin) -伊皮魯冰心(Epirubicin) -海左(Hydrea) 81 200835507Table 3: Chemotherapeutic agent -13-cis-vitamin A-2-amino-6-indenyl 嗓令 -2-CdA -2-chlorodeoxygenate - 5-fluorourine.定-5-FU -6-TG -6-guanine-6-M-based 嘌呤-6-MP-Accutane - Actinomycin-D-ararimycin (Adriamycin) - Adrucil - Agrylin - Ala-Cort - Aldesleukin - Alemtuzumab - Alitretinoin - Akaban (Alkaban)-AQ _ Alkeran - All-trans retinoic acid-α interferon - Altretamine - Amethopterin - Amifostine - A Aminoglutethimide - Anagrelide - Anandron - Anastrozole - arabinosyl-Ara-C - Aranesp - Aridai (Aranesp) Aredia) - Arimidex - Aromasin - Arsenic - Neosar - Neulasta _ Neumega - Neupogen _ Nilandron - Nilutamide - Nitrogen mustard - Novaldex - Novantrone - Octreotide - Onco, Oscar • Oncovin - Ontak - Onxal - Oprevelkin - Orapred - Orasone - Oxaliplatin - Paclitaxel - Pamidronate - Panretin - Pa Paraplatin - Pediapred - PEG interferon - Pegaspargase - Pegfilgrastim - PEG-INTRON - PEG-L-aspartame -Phenylmalonic acid mustard gas - Platinol - Platino - AQ - Prednisolone - Prednisone - Prelone - Procarbazine -PROCRIT 79 200835507 - Aspartate - ATRA - Avastin - BCG - BCNU - Bevacizumab - Bexarotene - Bicalutamide -BiCNU - Blenoxane - Bleomycin - Bortezomib - Busulfan - Busulfex - C225 - Leucovorin i Camp-Camptosar - Camptothecin-11 - Capecitabine - Carac - Carboplatin - Cardustin (Carmustine) - Casodex - CCNU -CDDP - CeeNU - Cembidine • cetuximab • Chlorambucil - Western Uranium 'Ting ( Cisplatin) - Citrovorum Factor - Cladribine - Cortisone - Cosmegen - CPT-11 - Cyclophosphamide - Saita Zourne Cytadren)_ - Proleukin - Prolifeprospan with Carmustine implants 20 - Purinethol • Raloxifene - Rumah (Rheumatrex) - Rituxan - Rituximab - Roveron - A (interferon a - 2a) - Rubex - Rubidomycin - Sandostatin - Alexandra LAR - Sargramostim - Solu-Cortef - S〇lu-Medrof - STI -571 - Streptozocin - Tamoxifen • Targretin - Taxol - Taxotere - Temodar - Timothy Temozolomide - Tuniposid e) -TESPA - Thalidomide - Thalomid - TheraCys - thioguanine - sulphur bird sputum; - Thiophosphoamide - thiopurine J ( Thioplex) - Thiotepa - TICE - Toposar - Topotecan - Toremifene - Trastuzumab _ 80 200835507 - Saitalabin (Cytarabine) - Tretinoin - 赛塔拉宾微脂粒-Trexall - Cytosar - U _ Trisenox - Cytoxan - TSPA - Dacarbazine - VCR - Dactinomycin - Velban _ Darbepoetin α - Vicade - Daunomycin - Wipxi (VePesid) - Daunorubicin - Vesanoid - Daunuo Bing Liquor - Viadul - DaunoXome - Vinblastine Sulfate ) - Decadron - Vincasar Pfs - Delta-Cortef - Vincristine - Deltasone - Vinorelbine - Dairyukin wears Diftitox - DepoCyt - VLB - Pexamethasone - VP · 16 - Desamethasone - Vumon - Desamethasone Sodium Phosphate - Xeloda - Desasone • Zanosar - Dexrazoxane - Zevalin - DHAD - Zinecard - DIC - Zola (Zoladex) - Diodex - Zoledronic acid - Docetaxel - Zometa - Doxil - Gliadel paste - Dorothy Ice Heart (Doxorubicin) - Glivec - Dossolu Ice Heart Lipid - GM-CSF - Prosy • Goserelin - DTIC • Granular Cell - Community Stimulating Factor - DTIC - Dome - Granulocyte - Macrophage - Community Stimulating Factor - Duralone - Halotestin - EfUdex - Herceptin - Eli嘉 ligard) - Hexadrol - Ellence - Hexalen - Eloxatin - hexamethyl melamine - Elspar - HMM • Ann Emcyt - Hycamtin - Piru Bing Xin (Epirubicin) - Sea left (Hydrea) 81 200835507

-伊普丁 (Epoetin) α -乙酸經可體(Hydrocort) -爾畢妥(Erbitux) -經可體松(Hydrocortisone) -艾文尼亞(Erwinia)L-天冬Si胺酶 -磷酸羥可體松鈉 -伊塔幕斯丁 (Estramustine) -丁二酸羥可體松鈉 -伊賽歐(；Ethyol) -填酸經可酮(Hydrocortone) -伊妥普福(Etopophos) -經基脲(Hydroxyurea) -伊妥普賽(Etoposide) -伊布土莫馬(Ibritumomab) -填酸伊女普赛 -土西坦(Tiuxetan) _優列辛(Eulexin) -伊達黴素(Idamycin) -艾文史塔(Evista) -伊達魯冰心(Idarubicin) -伊淫米史坦(Exemestane) -伊福(Ifex) -法列史東(Fareston) -IFN-a -法史羅戴(Faslodex) -伊福法邁(Ifosfamide) -費馬拉(Femara) -IL-2 -費葛拉斯堤(Filgrastim) -IL-11 -福羅蘇瑞丁 (Floxmidine) -甲石黃酸伊馬堤尼(Imatinib) -福達拉(Fludara) -咪唑羧醯胺 -福達拉賓(Fludarabine) -干擾素α -福羅普列(Fluoroplex) -干擾素a-2b(PEG軛合物） -氟尿嘴咬 -介白素-2 -氟尿嘧咬(乳膏） -介白素-11 -福赛米史特隆(Fluoxymesterone) -因壯(Intron)A(干擾素 a -2b) -福塔邁(Fhitamide) -魯可福林(Leucovorin) -酸葉酸(Folinic Acid) _魯克蘭(Leukeran) -FUDR -魯凱恩(Leukine) -福維斯特拉(Ful vestrant) -魯普萊(Leuprolide) -G-CSF -魯羅克里斯丁 (Leurocristine) -傑費堤尼(Gefitinib) -魯史塔丁 (Leustatin) -傑西塔賓(Gemcitabine) -微脂粒Ara-C -傑土竹美(Gemtuzumab)歐索嘉米辛 -液體普來德(Pred) (ozogamicin) -羅幕斯丁 (Lomustine) -傑薩(Gemzar) -L-PAM -葛列費(Gleevec) -L-莎可萊辛(Sarcolysin) -魯普(Lupron) -米堤可坦(Meticorten) -長效戴普(Dq)〇t) -米妥黴素(Mitomycin) -馬土蘭(Matulane) -米妥黴素-c _ 馬西戴(Maxidex) -米妥山特隆(Mitoxantrone) -美可羅薩敏(Mechlorethamine) -Μ-普尼索(Prednisol) -鹽酸美可羅薩敏 -MTC 82 200835507 -美拉隆(Medralone) -MTX _ 美左(Medrol) 幕斯塔金〇VTustarger〇 -美嘉斯(Megace) -幕斯丁 (Mustine) -美嘉斯左(Megestrol) •幕塔黴素(Mutamycin) -乙酸美嘉斯左 -邁蘭(Myleran) •美法蘭(Melphalan) -艾瑞莎(Iressa) -魏基嘌呤 -伊瑞諾堤坎(Irinotecan) -美納(Mesna) -伊索特堤諾(Isotretinoin) -美:$(Mesnex) -吉左拉斯(Kidrolase) -甲胺嗓呤(Methotrexate) _拉那可(Lanacort) -甲胺喋呤鈉 _L-天冬醯胺酶 -甲基普尼松隆(Methylprednisolone) -LCR -邁羅赛(Mylocel) -利妥左(Letrozole) 於若干實施例中，干擾素(IFN)可組合本發明化合物使用。適當干擾素包括：干擾素a_2a、干擾素α-2b、PEG化干擾素α包括干擾素a -2a及干擾素α _2b、干擾素/3、干擾 5素冷、干擾素Γ、干擾素ω、因法金(INFERGEN)(干擾素 a con-1)因特幕公司（interMune)製造、歐尼費隆 (OMNIFERON)(天然干擾素）維拉金公司（viragen)製造、歐布費隆（ALBUFERON)人基因體科學公司（H疆an Gen〇me Sciences)製造、瑞畢福(REBIF)(干擾素-ia)艾瑞斯-色隆 10諾公司（Ares-Senmo)製造、ω干擾素生醫公司（BioMedicine) 製造、口服干擾素α亞馬雷洛生科公司（Amarill〇 Biosciences)製造及干擾素τ、干擾素7、及/或干擾素 /3因特幕公司製造。於一個實施例中，如此處揭示iTCN、TCN-P或相關 15化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可與額外化學治療劑諸如此處所述或表3所示之額外 83 200835507 化學治療劑組合使用或交替使用來治療抗藥性癌症例如多重抗藥性癌症。抗藥性癌症包括大腸癌、骨癌、腎癌、腎上腺癌、胰癌、肝癌、及/或任何其它技藝界已知或如此處揭示之癌症。於一個實施例中，額外化學治療劑為P_糖蛋 5白抑制劑。於若干非限制性實施例中，P-糖蛋白抑制劑可選自於下列藥物：維拉帕米（verapamil)、賽羅史寶靈 (cyclosporin)(諸如賽羅史寶靈a)、塔莫西芬、鈣調節素 (calmodulin)拮抗劑、戴斯維拉帕米（dexverapamii)、戴尼谷戴平(dexniguldipine)、凡史普達(valspodar) (PSC 833)、拜 10 瑞可達（biricodar) (VX-710)、塔瑞奎達（tariquidar) (XR9576)、左蘇奎達（zosuquidar) (LY335979)、拉尼奎達 (laniquidar) (R101933)及/或ONT-093。 7·藥學組成物包括曲西立濱化合物及爾洛堤尼系列化合物例如傑費 15 堤尼、爾洛堤尼或其鹽之組成物視需要可與藥學上可接受之載劑或賦形劑投予。適合投予如此處提供之化合物之藥學上可接受之載劑包括熟諳技藝人士已知適合用於特定投藥模式之任何此等載劑。曲西立濱化合物且與爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可組合調配成為 2〇組成物中之唯一藥學活性成分或可組合調配。包括曲西立濱化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽之組成物適合用於經口、經直腸、經鼻、經局部（包括頰用及舌下）、經***或經腸道外（包括皮下、肌肉、皮下、靜脈、皮内、眼内、支氣管内、腦池 84 200835507 内、腹内、及硬膜外)投藥。較佳組成物係經靜脈投藥。組成物可方便地呈單位劑型呈現，或可藉習知製藥技術製備。此等技術包括將一種或多種本發明組成物與一種或多種藥學載劑或賦形劑組合之步驟。 5 曲西立濱化合物及爾洛堤尼系列化合物例如傑費堤尼爾洛立疋尼或其鹽及其組成物可調配成適當藥學製劑，諸如溶液劑、懸浮液劑、錠劑、分散錠劑、丸劑、膠囊劑、散劑、持續釋放配方、或醜劑供經口投藥；或可呈無菌溶液劑或懸浮液劑供腸道外投藥；以及經皮貼片製劑域粉 10吸入劑。於-個實施例中，前述曲西立濱化合物可使用技藝界眾所周知之技術及程序而調配成藥學組成物（例如參考Ansel，藥學劑型導論，第4版，1985年，126頁於該組成物中，有效濃度之一種或多種化合物或其藥學上可接叉之何生物可與一種或多種適當藥學載劑混合。 15本發明化合物可於調配前衍生成相對應之鹽類、醋類、稀醇賴或烯醇賴、_類、縮_、原酸賴、半祕類、半縮_類、酸類、鹼類、溶劑合物類、水合物類、或前藥。組成物中之化合物濃度可於投藥時有效遞送可治療、預防或改善目標疾病或病症之_種或多種症狀之用 2〇 Ϊ。於-個實施例中，組成物調配供單劑投藥。為了調配組成物，化合物之重量分量係以有效濃度溶解、懸浮、分散或以其它方式混合於所選之載劑，讓所治療的病情被緩解、預防、或改善一種或多種症狀。適合口服投藥用組成物可呈分開單位，諸如但非限於 85 200835507 錠劑、橢圓錠、丸劑、或糖衣劑、膠囊劑、或爲囊劑，其各自含預定量之一種或多種組成物；呈散劑或粒劑；呈於水性液體或非水性液體之溶液劑或懸浮液劑；或呈水包油型乳液劑或油包水型乳液劑或呈大丸藥等。 5 液體藥學可投予組成物之製法例如可經由將曲西立濱化合物及任選的藥學輔劑溶解、分散或以其它方式混合於載劑諸如水、食鹽水、水性葡萄糖、甘油、甘醇類、乙醇等，藉此溶解溶液或懸浮液。若有所需，欲投藥之藥學組成物也可含有小量無毒輔助性物質諸如濕潤劑、乳化劑、 10 增溶劑、pH緩衝劑、保藏劑、矯味劑等，例如乙酸鹽、擰檬酸鈉、環糊精衍生物、一月桂酸山梨聚糖酯、三乙醇胺乙酸鈉、三乙醇胺油酸醋、及其它此等輔劑。此種劑型之製法為熟諳技藝人士所已知或顯然易知例如參考雷明頓製藥科學，默克出版公司，賓州伊士頓。 15 適合局部投予口部之本發明組成物例如包括***錠，其含有各成分於矯味基劑通常為蔗糖及金合歡膠或西黃蓍膠；軟錠劑，其含有一種或多種本發明之曲西立濱化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽於惰性基劑諸如明膠及甘油或嚴糖及金合歡膠；及漱口藥， 20 其含有一種或多種本發明之組成物於適當液體載劑投予。鍵劑、丸劑、膠囊劑、喉片等可含有一種或多種具有類似性質之下列成分或化合物：黏結劑；潤滑劑；稀釋劑；滑動劑；崩散劑；著色劑；甜味劑；矯味劑；濕潤劑；催吐包衣；及膜衣。黏結劑之實例包括微晶纖維素、西黃蓍 86 200835507 膠、葡萄糖溶液、金合歡黏質、明膠溶液、糖蜜、聚乙烯基吡咯啶酮、普維隆(povidone)、交聯普維隆、蔗糖及澱粉糊。潤滑劑包括滑石、澱粉、硬脂酸鎂或硬脂酸鈣、萊可普登(lycopodium)、及硬脂酸。稀釋劑例如包括乳糖、蔗糖、 5澱粉、高嶺土、鹽、甘露糖醇、及磷酸二鈣。滑動劑包括但非限於膠體二氧化矽。崩散劑包括交聯甲基纖維素鈉、乙醇酸澱粉鈉、褐藻酸、玉米澱粉、馬鈴薯殿粉、膨潤土、甲基纖維素、填脂、及魏甲基纖維素。著色劑例如包括經核准及許可之水溶性FD&C染料中之任一者、其混合物；及 10懸浮於水合礬土上之水不溶性FD&C染料。甜味劑包括蔗糖、乳糖、甘露糖醇、及人工甜味劑諸如糖精及喷乾矯味劑中之任一者。矯味劑包括萃取自植物諸如水果之矯味劑’其可產生怡人感覺之化合物之合成摻合物，諸如但非限於薄荷腦及水楊酸甲酯。濕潤劑包括一硬脂酸丙二醇 15 酯、一油酸山梨糖醇酯、一月桂酸二乙二醇酯及聚氧伸乙基月桂基醚。催吐包衣包括脂肪酸類、脂肪類、蠟類、蟲膠、氨化蟲膠、及乙酸磷苯二曱酸纖維素。膜衣包括羥乙基纖維素、羥甲基纖維素鈉、聚乙二醇4〇〇〇及乙酸磷苯二甲酸纖維素。 20 適合局部投予皮膚之組成物可呈有一種或多種組成物於藥學上可接受之載劑投予之軟膏劑、乳膏劑、凝膠劑、及糊劑劑型。直腸投藥用組成物可呈含適當基劑例如包括可可脂或水楊酸酯之栓劑劑型投予。 87 200835507 適合供經鼻投藥之組成物，當載劑為固體時，組成物包括具有20微米至500微米範圍之粒徑之粗粉，以呈鼻嗅粉之方式投藥（亦即將粉末容器保持接近鼻孔快速吸入通過鼻道投藥）。當載劑為液體(例如鼻噴霧劑或鼻滴劑）時，一 5 種或多種組成物可混合於水性溶液或油性溶液而吸入鼻道或喷霧進入鼻道。適合供***投藥之組成物可呈含有一種或多種組成物及適當載劑之子宮托、棉塞、軟膏劑、明膠劑、糊劑、發泡劑或喷霧配方劑型。 10 適合供腸道外投藥之組成物包括水性及非水性無菌注射溶液劑，其可含有抗氧化劑、緩衝劑、制菌劑、及讓配方變成與期望接受者之血液呈等張性之溶質，及水性無菌懸浮液劑及非水性無菌懸浮液劑，其包括懸浮劑及增稠劑。組成物可呈單劑或多劑容器例如密封安瓿及密封小瓶 15 形式，組成物可儲存於冷凍乾燥（凍乾)條件下，只需恰在使用之前添加無菌液體載劑例如注射用水。臨時注射溶液劑及懸浮液劑可由前文說明之該種無菌散劑、粒劑及錠劑製備。適合供腸道投藥或腸道外投藥之藥用有機或無機固體 20 或液體載劑介質可用來製造組成物。明膠、乳糖、澱粉、硬脂酸鎭、滑石、植物及動物脂肪類及油類、樹膠、聚伸烷基二醇、水、或其它已知載劑全部皆適合用作為載劑介質。包括曲西立濱化合物及爾洛堤尼系列化合物例如傑費 88 200835507 支疋尼、爾洛堤尼或其鹽之組成物可組合〜可接党之载劑介質及/或賦形劑使用 ’夕種糸學上上可接受之_介質」包肺-觀全「藥學釋劑、或以其它液體載媒劑、分散助 1〜卜稀活性劑、等_、增_或乳化劑、保表面潤滑劑、_、«劑、細統、固體黏結劑、藏劑、界面活性劑、著色劑、橋味劑义^ 適合用於期望之特定劑型而使用。_味劑等皆依其 10 15 此外，包括曲西立濱化合物及爾洛如傑費堤尼、爾洛堤尼或其鹽之組成物可：= 之賦形劑及任選地持續釋放基體諸如可 /、予σ文合來形絲雜組成物。「鮮上可錢2分解聚合物組母固體、半固體或㈣過㈣、稀釋囊二括無類型之配方_。細材料或任何較但須瞭解組成物之總每日使用係由臨床醫師依據完整醫療判定之範圍決定。用於任何特定宿主之特定治療有2 二將依據多·素決定，例如包括所治療之病症及該病症嚴重程度；所使用之特定組成物之活性；所使用之特定2 成物、病人年齡、體重、一般健康狀況、性別及飲食；投藥時間；投藥途徑；所使用之特定化合物之***速率；治療時間；與所採用之特定組成物組合使用或同時使用之曲西立濱化合物及/或爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽等藥物業界眾所周知之因素。舉例言之，於業界人士之技巧範圍内已知以比達成期望療效所需劑量更 20 200835507 量開始投予該組成物，以及徐緩提高劑量至達到期包括曲西立濱化合物及爾洛堤尼系列化合物例堤尼、爾洛堤尼或其鹽之組成物為了容易投藥及劑量的均一，較佳刺喊單位㈣。如此處❹之「單位係指適合驗欲治療之宿主之組成物之實體上分料位」。各劑量須含有計算來就此、或於所選藥學上制介質結人來產生期望之療效之組成物數量。、 10 15 車^單位劑量配方為含有所投予成分之每日劑量或每 “早位、母曰次劑量或其適當分量之單位劑。 =如母日約Μ毫克此處揭示之化合物可缩小小鼠體實體腫瘤體積。 _ 之劑量將依據宿主因素諸如體重、年組織分布、魏速率及***鱗決H個實施^射、每曰約0.5克至7克如此處揭示之曲西立物，類。任選地，每日約口物可投予人於若干W 濱化合物可投予人類。於右干貫瓠例中，每曰約〇·_毫克至5毫 =:如療有效劑量將依據前文說明之二、心卜’於技#界技巧範圍内可於相予組成物1及提高劑量至達到期望的效果。· 4始技 ^包=西立濱化合物及爾洛堤尼系列化合物例如傑費 :可由可Γ/是尼或其鹽之組成物可用於持續釋放基體，基 ' _酵素水解或基於酸之水解或藉溶解>_#14 製成，”為聚合物一身雜内部，基體= 20 200835507 體液的作用。持續釋放基體例如係選自於可生物相容之材 ”微脂粒類、聚丙μ旨類（聚乳酸）、聚乙μ旨（乙醇暖聚合物)、聚丙交自旨共聚合乙交酷(乳酸與乙醇酸之共： ^聚軒類、聚(原酸)醋類、多月太類、玻尿酸、膠原蛋白二瓜㈣人骨素、叛酸類、脂肪酸類、填脂類 g音、夕 /呢撕核酸、夕胺基酸類、胺基酸類諸如苯基丙胺酸、㈣酸、里白胺酸、多核細、聚乙稀基丙烯、聚乙稀基吡咯啶_、二=料。較佳可生物分解基體為聚丙交§旨、聚乙交喂、 1或ΛΚ丙父s旨共聚合乙交醋（乳酸與乙醇酸之共聚物）中之— 1〇者之基體。曲西立濱化合物及爾洛堤尼系列化合物例如傑費护尼、爾洛堤尼或其鹽也可呈微脂粒形式投予。如技藝界k U月曰粒通常係衍生自碟脂或其它脂質物質。微骑由單層或多層水合液晶分散於水性介質所形成。可使’、 15 =成微絲之任-種無毒性生理上可接受性且可代謝之月: 貝除了本發明之一種或多種組成物外，微脂粒可含有6 疋劑、保藏劑、賦形劑等。脂質之實例為天然及合成之: 脂類及石粦脂基膽驗類（卵鱗脂類）。微脂粒之形成方法為H 界眾所周知。〜、技藝 20 曲西立濱化合物及爾洛堤尼系列化合物例如傑費抒尼、爾洛堤尼或其鹽可調配成氣溶膠供例如藉吸入方式= 用投予呼吸逼之此等配方可呈氣溶膠或噴霧劑溶液:式或呈U細。人入用粉末形式，可單獨使用或組合惰性栽劑^ 如乳糖使用。於此種情況下，於-個實施例中，配方二粒 200835507 5 10 15 20 具有小於5G微米之直徑，於-個實施例巾小則峨米。包括曲西立濱化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽之組成物可與其它治療前述病情之組成物及/絲雜合使用。舉财之，種瘤f知係使用手術、放射性治療或化學治療組合-種或多種本發明組成物一，隨後-種或多種本發明組成物可投予該病人來延長微小遽瘤轉移的休眠’且穩定、抑制或減少任何殘留原發性腫瘤的生長。 X 7.h額外實施例包括曲西立濱化合物及爾洛堤尼系列化合物例堤尼、爾洛堤尼或其鹽之藥學組成物可根據已知= 有用組之製法概。㈣制於絲 = 知且方便易得之多個來源士眾所周«e古八η — " 田明頓製藥科學， ^克出版“’㈣伊均說明可關聯方。適合投予之配方例如包括水性及非水性益=之配 :’其可含有抗氧化劑、緩衝劑、配二液與期望接受者之血液呈等張性之溶f方變成劑及非水性無菌懸浮液無囷懸浮液可呈單劑或多劑容器例二;包括懸浮劑及增稠劑。配方物可儲存於冷;東乾燥咏^ h瓶及讀小瓶形式，級成無菌液體載劑例如注射用c条件下’只需恰在使用之前添加可由無菌散劑、粒劑、餘:臨時注射溶液劑及懸浮液劑說明之成分之外，本發明=製備。須瞭解除了前文特別藝界習知之其它化學劑/方可包括有__配方之技處理 92 200835507 5 10 15 20 本發明方法例如用於抑制癌性細胞的生長，本發明方法較佳組合至少另-種治療方法，包括但非限於化學治療、放射性治療、選擇性抑制Ras致癌基因發訊之治療、或任何其它熟諳癌症處理及處置技藝界人士已知之治療，^ 如投予抗癌劑。 = 也可進行投予呈鹽之API_2 (曲西立濱）。藥學上可接典之鹽之實例為與與可形成生理上可接受之陰離子之酸所= 成之有機酸加成鹽，例如甲笨俩鹽、甲俩鹽、乙酸臨^ 棒檬酸鹽、丙二酸鹽、酒石酸鹽、丁二酸鹽、苯甲酸趟现、抗壞血酸鹽、酮基戊二酸鹽、及甘油基碟酸臨^可形成適當之無機酸鹽包括硫酸鹽、硝酸鹽、重碳酸趟碳酸鹽。 1久轉上可触之鹽切㈣技藝界眾所料之標準程又传’例如充分祕化合物諸如贿適# =之陰離子之酸反應。也可製繼之驗金屬= ^鉀或鋰）鹽或鹼土金屬（例如鈣）鹽。曲西立濱化合物及爾洛堤尼系列化合物例如傑費户匕、爾洛堤尼或其鹽可調配成藥、疋投藥形式亦即經口或經腸道外，、靜1 1選用之次皮下㈣投予個體諸如病人或生病動物。物例如俾費=明之曲西立濱化合物及爾洛堤尼系列化合媒劑(亦=、爾，性投藥。可^壯為如^性稀釋劍或可同化食用載劑經系統封衣於硬殼紐殼明膠膠囊内，可壓縮成為旋 93 200835507 劑或可直接與病人食物摻混。供口服治療投藥用，該等化合物組合一種或多種賦形劑，且以可攝食錠劑、頰用錠、喉片、膠囊劑、酏劑、懸浮液劑、糖漿劑、糊劑等劑型使用。此等組成物及製劑須含有至少0.1%活性劑。組成物及 5 製劑之百分比當然可改變，方便地係占給定單位劑型重量之約2%至約60%重量比之範圍。於此種治療有用之組成物中之活性化合物含量為可獲得有效劑量水平。錠劑、喉片、丸劑、膠囊劑等也含有下列成分：黏結劑諸如西黃蓍膠、金合歡膠、玉米澱粉或明膠；賦形劑諸 10 如磷酸二鈣；崩散劑諸如玉米澱粉、馬鈐薯澱粉、褐藻酸等；潤滑劑諸如硬脂酸鎂；及甜味劑諸如嚴糖、果糖、乳糖或阿斯巴甜；及矯味劑諸如薄荷腦、冬綠油或櫻桃口味。當單位劑型為膠囊劑時，除了前述類別之材料之外，膠囊劑含有液體載劑諸如植物油或聚乙二醇。可存在有多種其 15 它形式作為包衣，或以其它方式來修飾固體單位劑型的實體形狀。例如，錠劑、丸劑或膠囊劑可以明劑、蠟、蟲膠或糖等包衣。糖漿劑或酏劑可含有本發明化合物，蔗糖或果糖作為甜味劑，對羥基苯曱酸甲酯及對羥基苯甲酸丙酯作為保藏劑，染料及繞味劑諸如櫻桃口味或柳撥口味。當 20 然用於製備任何單位劑型之任何材料須為藥學上可接受性且於其使用量為實質上無毒。此外，本發明化合物可結合入持續釋放製劑及裝置。曲西立濱化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽也可藉輸注或藉注射經靜脈投予或腹 94 200835507 技予。活性劑或其鹽之溶液可界面活性㈣i 要可此合無毒備也可於甘油、賴聚乙二醇、 5 10 15 20 二=物製備’及於油製備。於尋常儲錢使此等製劑含有防止微生物生長之保藏劑。益菌主之_咖包括包含活性成分之二劑或水性分散液劑或無菌散劑，其適〜臺囷/射用溶_或輸注歸液劑或分散液劑，視泰件封於微脂粒。總而言之，最終劑型於製造與儲存：，為無菌、流體安定。液體載劑 , 或^分散介質包括例如水、乙醇、多元醇(例如甘油= 、卜、液體聚乙二義等）、植物油、無毒甘油醋類、及复 =田=合物。適當流體性質例如可經由微脂粒之形成，綾劑液之情況下維持所妹徑，或經由使科面活性、、隹持。藉各種抗菌劑及抗真菌劑可獲得微生物作用 :::例如對羥基苯曱酸酯類、氣丁醇、酚、山梨酸、柳南=等於夕種情況下，較佳含括等張劑例如糖類、緩衝 2氯化鈉。注射用組成物可藉由於組成物中使用延遲吸蜊例如一硬脂酸鋁及明膠來獲得延長吸收。旦無菌注射溶液劑之製法係經由將本發明化合物以需要 ^見需要於前述多種成分摻混於適t溶劑内，接著過:滅 =。於用於製備錢注溶㈣之無®散劑之情況下，卜方法為真空乾_及冷'東乾燥技術，獲得活性成分加任 :碩外存在於前述經過錢過渡之溶液中之期望成分之散片J ο 95 200835507 供局部投藥用，曲西立濱化合物及爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽可以純質形式使用，亦即呈液體。但通常期望組合皮膚可接受之載劑，載劑可為固體或液體而呈組成物或配方施用於皮膚。 5 有用之固體載劑包括細分固體，諸如滑石、黏土、微晶纖維素、矽氧、鋁氧等。有用之液體載劑包括水、醇類或甘醇類或水-醇/甘醇摻合物，其中本發明化合物視需要可藉助於無毒性界面活性劑而以有效含量溶解或分散。可添加輔劑諸如香水及額外抗微生物劑來對一給定之用途獲得 10 最佳性質。所得液體組成物可由吸收墊施用，用於浸潰繃帶及其它敷料，或使用幫浦型喷灑器或氣溶膠喷灑器而噴灑於患部區。增稠劑諸如合成聚合物類、脂肪酸類、脂肪酸鹽酸、及脂肪酸酯類、脂肪醇類、改性纖維素類、或改性礦物材 15料也可用於液體載劑來形成可展開糊料、凝膠劑、軟膏劑、息劑等供直接施用於使用者皮膚。可用來將本發明化合物遞送至皮膚之有用的皮膚用組成物之實例係揭示於Jacquet 等人(美國專利案4,608,392)、Geria(美國專利案4,992,478)、 Smith等人（美國專利案4,559 157)&w〇ltzman (美國專利案 20 4,820,508)。本發明之藥學組成物之有用劑量可經由比較其於試管内之活性及於動物研究模型中之活體内活性來測定。於小乳及其它動物之有效劑量外推至人類之方法為技藝界所已知；例如參考美國專利案4,938,949。 96 200835507 於一個非限制性實施例中，液體組成物諸如洗劑中之活性劑濃度可為約0.1-25 wt·-%或由約0.5-10 wt·-%。於一個貝％例中，於半固體組成物或固體組成物諸如凝膠劑或散 ^中之’辰度約為〇·1-5 wt._%，較佳約為0.5-2.5 wt·-%。於一 5们貫加例中，〉主射、輪注或攝食用之單一劑量通常為5-1500 *克，可每日投予U次來於成人獲得約0.1-50毫克/千克之 /辰度本發明之非限制性劑量為每曰7.5毫克至45毫克口服投藥，對個人體重可做適當調整。- Epoetin alpha-acetic acid Hydrocort - Erbitux - Hydrocortisone - Erwinia L-aspartate Siamine - Hydroxy phosphate Estramustine - Sodium succinate-Essex (Ethyol) - Hydrocortone - Etopophos - urethraquinone (Etopophos) Hydroxyurea) - Etoposide - Ibritumomab - Tised acid - Tixitan _ Eulexin - Idamycin - Ai Wenshi Evista - Idarubicin - Exemestane - Ifex - Fareston - IFN-a - Faslodex - Ilfafa ( Ifosfamide) - Femara - IL-2 - Filgrastim - IL-11 - Floxmidine - Imatinib - Fudala Fludara) -Imidazole Carboxamide -Fudadarabine -Interferon alpha -Fooroplex -Interferon a-2b (PEG conjugate) -Fluorine bite -Interleukin-2 - Fluorouracil bite (cream) - Interleukin-11 - Forsyme history Fluoxymesterone - Intron A (interferon a -2b) - Fhitamide - Leucovorin - Folinic Acid _ Leukeran - FUDR - Leukine - Ful vestrant - Leuprolide - G-CSF - Lerocristine - Gefitinib - Rustaldine (Leustatin) - Gemcitabine - Alipid Ara-C - Gemtuzumab Osogamisin - Liquid Pred (ozdamicin) - Lomustine - Gemzar -L-PAM -Gleevec -L-Sarcolysin -Lupron -Meticorten -Long-term Dyke (Dq)〇t ) - Mitomycin - Matulane - Mirtamycin - c _ Maxidex - Mitoxantrone - Mechlorethamine - Μ - Prednisol - Mecorosamin Hydrochloride - MTC 82 200835507 - Medralone - MTX _ Medrol 斯塔〇〇 VTustarger〇-Megace - Mustin Mustine) - Megestrol • Moutotamycin (Mut Amycin) - Myleran Acetate • Melphalan - Iressa - Irinotecan - Mesna - Isot Isotretinoin - US: $(Mesnex) - Kidrolase - Methotrexate _ Lanacort - Methylammonium _L-aspartate - Methylprednisolone - LCR - Mylocel - Letrozole In several embodiments, interferon (IFN) can be used in combination with a compound of the invention. Suitable interferons include: interferon a_2a, interferon alpha-2b, pegylated interferon alpha including interferon a-2a and interferon alpha _2b, interferon/3, interferon 5 cold, interferon Γ, interferon ω, INFERGEN (interferon a con-1) manufactured by interMune, OMNIFERON (natural interferon) manufactured by Viragen, and Obufferon (ALBUFERON) Manufactured by Human Genome Sciences (H Xinjiang an Gen〇me Sciences), REBIF (Interferon-ia) manufactured by Ares-Senmo, Omega Interferon The company (BioMedicine) manufactures, orally manufactures interferon alpha Amarillo Biosciences and interferon τ, interferon 7, and/or interferon/3. In one embodiment, the iTCN, TCN-P or related 15 compound and the erlotini series of compounds such as gefitinib, erlotinib or a salt thereof can be disclosed herein with additional chemotherapeutic agents such as described herein or The additional 83 200835507 chemotherapeutic agents shown in Table 3 are used in combination or alternately to treat drug-resistant cancers such as multi-drug resistant cancers. Drug-resistant cancers include colorectal cancer, bone cancer, kidney cancer, adrenal cancer, pancreatic cancer, liver cancer, and/or any other cancer known to the artisan or as disclosed herein. In one embodiment, the additional chemotherapeutic agent is a P_sugar egg white inhibitor. In several non-limiting embodiments, the P-glycoprotein inhibitor can be selected from the group consisting of verapamil, cyclosporin (such as celathlon a), tamaxifen, calcium. Calmodulin antagonist, dexverapamii, dexniguldipine, valspodar (PSC 833), and biricodar (VX-710) , tariquidar (XR9576), zosuquidar (LY335979), laniquidar (R101933) and/or ONT-093. 7. A pharmaceutical composition comprising a composition of a triclinide compound and a erlotinib compound such as gefe 15 tiani, erlotini or a salt thereof, optionally with a pharmaceutically acceptable carrier or excipient Cast. Pharmaceutically acceptable carriers suitable for administration of the compounds provided herein include any such carriers known to those skilled in the art to be suitable for the particular mode of administration. The triclinide compound and the erlotiniline compound such as gefitinib, erlotinib or a salt thereof may be formulated in combination to form the sole pharmaceutically active ingredient in the bismuth composition or may be formulated in combination. Compositions including trifluraline compounds and erlotini series compounds such as gefitini, erlotini or their salts are suitable for oral, rectal, nasal, and topical (including buccal and sublingual) ), transvaginal or parenteral (including subcutaneous, intramuscular, subcutaneous, intravenous, intradermal, intraocular, intrabronchial, cerebral cistern 84 200835507, intra-abdominal, and epidural). Preferred compositions are administered intravenously. The compositions may conveniently be presented in unit dosage form or may be prepared by conventional pharmaceutical techniques. Such techniques include the step of combining one or more of the compositions of the present invention with one or more pharmaceutical carriers or excipients. 5 Qucilide compound and erlotini series of compounds such as gefetilil lorinib or its salt and its composition can be formulated into appropriate pharmaceutical preparations, such as solutions, suspensions, lozenges, dispersions The agent, the pill, the capsule, the powder, the sustained release formula, or the ugly agent for oral administration; or the sterile solution or suspension for parenteral administration; and the transdermal patch preparation powder 10 inhalation. In one embodiment, the aforementioned triclinide compound can be formulated into a pharmaceutical composition using techniques and procedures well known in the art (for example, refer to Ansel, Introduction to Pharmaceutical Formulations, 4th Ed., 1985, p. 126). Wherein the effective concentration of one or more compounds or pharmaceutically acceptable organisms thereof may be combined with one or more suitable pharmaceutical carriers. 15 The compounds of the invention may be derivatized into corresponding salts, vinegars, and diluents prior to formulation. Alcohol lysine or enol lysine, _ class, condensed _, ortho sulphate, semi-secret, semi-condensed, acid, base, solvate, hydrate, or prodrug. The drug can be effectively delivered at the time of administration to treat, prevent or ameliorate the symptoms or symptoms of the target disease or condition. In one embodiment, the composition is formulated for single administration. To formulate the composition, the compound The weight component is dissolved, suspended, dispersed or otherwise mixed at a concentration effective to the selected carrier to allow the condition to be treated to be alleviated, prevented, or ameliorated by one or more symptoms. May be in separate units such as, but not limited to, 85 200835507 tablets, elliptical tablets, pills, or dragees, capsules, or sachets, each containing a predetermined amount of one or more of the compositions; in the form of a powder or granule; a solution or suspension of an aqueous liquid or a non-aqueous liquid; or an oil-in-water emulsion or a water-in-oil emulsion or a bolus or the like. 5 A method for preparing a liquid pharmaceutically acceptable composition, for example, The triclinide compound and the optional pharmaceutical adjuvant are dissolved, dispersed or otherwise mixed in a carrier such as water, saline, aqueous dextrose, glycerol, glycols, ethanol, etc., thereby dissolving the solution or suspension. If desired, the pharmaceutical composition to be administered may also contain small amounts of non-toxic auxiliary substances such as wetting agents, emulsifiers, 10 solubilizers, pH buffering agents, preservatives, flavoring agents, etc., such as acetate, sodium citrate, Cyclodextrin derivatives, sorbitan monolaurate, sodium triethanolamine acetate, triethanolamine oleic acid vinegar, and other such adjuvants. The preparation of such dosage forms is known or apparent to those skilled in the art. See, for example, Remington Pharmaceutical Sciences, Merck Publishing Company, Easton, Pennsylvania. 15 Compositions of the invention suitable for topical administration to the mouth include, for example, buccal ingots containing various ingredients in a flavoring base, typically sucrose and acacia a gum or tragacanth; a soft lozenge containing one or more of the triclinic compound of the present invention and a compound of erlotini such as gefitini, erlotini or a salt thereof in an inert base such as gelatin And glycerin or sucrose and acacia gum; and mouthwash, 20 which comprises one or more of the compositions of the invention administered in a suitable liquid carrier. The granules, pills, capsules, throat tablets, etc. may contain one or more The following ingredients or compounds having similar properties: binder; lubricant; diluent; slip agent; disintegrating agent; coloring agent; sweetener; flavoring agent; wetting agent; emetic coating; and film coating. Examples of bonding agent include Microcrystalline cellulose, scutellaria 86 200835507 Glue, glucose solution, acacia viscous, gelatin solution, molasses, polyvinylpyrrolidone, povidone, cross-linked Pirillon, sucrose and starch pasteLubricants include talc, starch, magnesium stearate or calcium stearate, lycopodium, and stearic acid. Diluents include, for example, lactose, sucrose, 5 starch, kaolin, salt, mannitol, and dicalcium phosphate. Sliding agents include, but are not limited to, colloidal cerium oxide. Disintegrating agents include crosslinked methylcellulose sodium, sodium starch glycolate, alginic acid, corn starch, potato powder, bentonite, methylcellulose, fat-filled, and Weimethylcellulose. The colorant includes, for example, any of the approved and approved water-soluble FD&C dyes, mixtures thereof; and 10 water-insoluble FD&C dyes suspended on hydrated alumina. Sweeteners include any of sucrose, lactose, mannitol, and artificial sweeteners such as saccharin and spray dry flavors. Flavoring agents include synthetic blends of compounds extracted from plants such as fruits, which produce a pleasant sensation, such as, but not limited to, menthol and methyl salicylate. The humectant includes propylene glycol stearate 15 ester, sorbitan monooleate, diethylene glycol monolaurate, and polyoxyethylene lauryl ether. The emetic coating includes fatty acids, fats, waxes, shellac, ammoniated shellac, and cellulose acetate phthalate. The film coat includes hydroxyethyl cellulose, sodium carboxymethyl cellulose, polyethylene glycol 4, and cellulose acetate phthalate. 20 A composition suitable for topical administration to the skin may be in the form of an ointment, cream, gel, and paste for which one or more of the compositions are administered in a pharmaceutically acceptable carrier. The rectal pharmaceutical compositions can be administered in the form of a suppository containing a suitable base such as cocoa butter or a salicylate. 87 200835507 Suitable for nasal administration of a composition, when the carrier is a solid, the composition comprises a coarse powder having a particle size ranging from 20 micrometers to 500 micrometers, which is administered as a nasal olfactory powder (ie, the powder container is kept close) Rapid inhalation of the nostrils through the nasal passages). When the carrier is a liquid (e.g., a nasal spray or a nasal drop), one or more of the compositions may be mixed with an aqueous solution or an oily solution to be inhaled into the nasal passages or sprayed into the nasal passages. Compositions suitable for vaginal administration may be presented as a pessary, tampons, ointment, gelatin, paste, foaming or spray formulation containing one or more compositions and suitable carriers. 10 Compositions suitable for parenteral administration include aqueous and non-aqueous sterile injectable solutions, which may contain antioxidants, buffers, bacteriostatic agents, and solutes which render the formulation is isotonic with the blood of the intended recipient, and aqueous sterility Suspension and non-aqueous sterile suspensions, including suspending agents and thickening agents. The composition may be in the form of a single or multiple dose containers such as sealed ampoules and sealed vials 15 and the composition may be stored under lyophilization (lyophilization) conditions by the addition of a sterile liquid carrier such as water for injection just prior to use. The temporary injectable solutions and suspensions can be prepared from the sterile powders, granules and lozenges described above. A medicinal organic or inorganic solid suitable for enteral or parenteral administration 20 or a liquid carrier medium can be used to make the composition. Gelatin, lactose, starch, barium stearate, talc, vegetable and animal fats and oils, gums, polyalkylene glycols, water, or other known carriers are all suitable for use as a carrier medium. Including the composition of the triclinide compound and the erlotini series of compounds such as Jeffrey 88 200835507, the composition of the sputum, erlotini or its salt can be combined ~ the carrier medium and / or excipients can be used夕糸上上」」」包包包包包包包包包包包包「「「「「「「「「「「「「「「「「「「「「「「「「「「「「「「 Lubricants, _, «agents, fines, solid binders, sizing agents, surfactants, colorants, bridge scent agents ^ are suitable for the specific dosage form desired. _ scent agents, etc. according to their 10 15 a composition comprising a triclinide compound and a composition such as erroprofen, erlotinib or a salt thereof, which may be: = an excipient and optionally a sustained release matrix such as a sigma Silk composition. "Freshly soluble 2 decomposition polymer group mother solid, semi-solid or (4) over (four), dilute capsules, including no type of formula _. Fine materials or any of the total daily use of the composition It is determined by the clinician based on the scope of the complete medical decision. It is used for any particular host. Treatment will be determined on the basis of multiple factors, including, for example, the condition being treated and the severity of the condition; the activity of the particular composition used; the specific 2 components used, the age, weight, general health, sex of the patient And diet; time of administration; route of administration; rate of excretion of a particular compound used; time of treatment; tresibine compound and/or erlotini series compound, such as jie, used in combination with or in combination with the particular composition employed Factini, erlotini or its salt are well known factors in the pharmaceutical industry. For example, within the skill of the industry, it is known to start administering the composition at a dose of 20,035,507, which is more than the dose required to achieve the desired therapeutic effect. And slowly increasing the dose to reach the stage including the composition of the triclinide compound and the erlotini series of compounds such as dynasty, erlotini or its salt, in order to facilitate the administration and uniformity of the dose, preferably the screaming unit (four). The term "unit means the physical partitioning of the composition of the host suitable for the treatment of the subject". Each dose must contain a quantity of the composition calculated to produce the desired therapeutic effect in this regard, or in the selected pharmaceutical medium. , 10 15 car ^ unit dose formula is the daily dose containing the ingredients to be administered or each "early dose, mother-in-law dose or its appropriate amount of unit agent. = If the mother day is about Μ mg of the compound disclosed here can be reduced The body volume of the mouse body tumor. The dose of _ will be based on host factors such as body weight, annual tissue distribution, Wei rate, and excretion scales. Each shot is about 0.5 grams to 7 grams per gram, as shown here. Optionally, a daily oral dose can be administered to a human in a number of W compounds to be administered to humans. In the case of the right sputum, each 曰·_mg to 5 milligrams =: According to the second description of the above, the heart of the 'Technology#' can be used in the composition of the composition 1 and increase the dose to achieve the desired effect. · 4 Start Technology ^ package = Cilibin compound and Lerotiini series of compounds For example, Jeffrey: can be used for the sustained release of the matrix, which can be used for the sustained release of the matrix, based on the hydrolysis of the enzyme or the hydrolysis based on acid or by the dissolution >_#14," , matrix = 20 200835507 The role of body fluids. The sustained release matrix is, for example, selected from the group consisting of biocompatible materials, microlipids, polyacrylic acid (polylactic acid), polyethylamine (ethanol warm polymer), and polyacrylic acid copolymerization (Ethylene glycol). A total of lactic acid and glycolic acid: ^ Juxuan, poly (original acid) vinegar, multi-monthly, hyaluronic acid, collagen, melon (four) human bone, tick, fatty acid, fat g, eve / ah Tear nucleic acid, oxime acid, amino acid such as phenylalanine, (tetra) acid, leucine, polynuclear fine, polyvinyl propylene, polyvinylpyrrolidine _, bis = material. The decomposed matrix is a matrix of the one of the ones of the co-polymerized vinegar (copolymer of lactic acid and glycolic acid), which is a combination of polyacrylic acid, polystyrene, or propylene. The Lotini series of compounds such as Jeffrey, erlotinib or a salt thereof can also be administered in the form of microlipids. For example, the kiln granules are usually derived from dish fat or other lipid substances. Single or multi-layer hydrated liquid crystals are dispersed in an aqueous medium to form ', 15 = micro-filaments - non-toxic physiological Acceptable and Metabolizable Months: In addition to one or more of the compositions of the present invention, the vesicles may contain 6 elixirs, preservatives, excipients, etc. Examples of lipids are natural and synthetic: lipids and The sarcophagus-based biliary test (oval squamous lipids). The formation of vesicles is well known in the H world. ~, skill 20 Quci Libin compounds and erlotini series compounds such as Jeffrey, Erlot Nitrogen or its salt can be formulated into an aerosol for use, for example, by inhalation = by breathing, such a formulation can be in the form of an aerosol or spray solution: or as fine as U. The powder can be used alone or in combination. Inert ingredients such as lactose are used. In this case, in one embodiment, the formula two tablets 200835507 5 10 15 20 has a diameter of less than 5G micrometers, and in one embodiment the towel is small in size. The composition of the Ribin compound and the erlotini series of compounds such as gefitini, erlotini or its salt can be used in combination with other compositions for treating the aforementioned conditions and/or silk. Use a combination of surgery, radiation therapy or chemotherapy - Or a plurality of compositions of the invention, followed by one or more of the compositions of the invention may be administered to the patient to prolong the dormancy of microtumor metastasis' and to stabilize, inhibit or reduce the growth of any residual primary tumor. X 7.h Additional examples include the composition of the triclinide compound and the erlotini series of compounds, such as dynasty, erlotinib or a salt thereof, according to the known method of the useful group. (4) Manufactured in silk = knowing and convenient Many sources of easy access to the Department of the People's Week Zhou «e ancient eight η — " Tian Mingdun Pharmaceutical Science, ^ gram published "'(four) Yi are indicative of related parties. Formulas suitable for investment include, for example, water-based and non-aqueous benefits = Formulation: 'It may contain an antioxidant, a buffer, a two-liquid solution and an isotonic solution of the blood of the intended recipient, and a non-aqueous sterile suspension, the sputum-free suspension may be in a single or multi-dose container; Including suspending agents and thickeners. The formula can be stored in cold; East dry 咏 ^ h bottle and read vial form, graded into a sterile liquid carrier such as injection c conditions - just add just before use, can be used as sterile powder, granules, and other: temporary injection solution In addition to the ingredients described for the agents and suspensions, the invention = preparation. It is to be understood that other chemical agents may be included in the art in addition to the prior art. The method of the present invention is for example, for inhibiting the growth of cancerous cells, and the method of the present invention preferably combines at least another A method of treatment, including, but not limited to, chemotherapy, radiation therapy, selective inhibition of Ras oncogene signaling, or any other treatment known to those skilled in the art of cancer treatment and disposal, such as administration of an anticancer agent. = API 2 (Quxi Libin) for salt administration can also be administered. Examples of pharmaceutically acceptable salts are organic acid addition salts with acids which form physiologically acceptable anions, such as methyl phenate, methyl salt, acetic acid, citrate, c. Diacid salts, tartrates, succinates, benzoic acid oxime, ascorbate, ketoglutarate, and glyceryl acid can form suitable mineral acid salts including sulfates, nitrates, and bicarbonate. Bismuth carbonate. 1 For a long time, the salty cut can be touched. (4) The standard course of the art industry is also known as the acid reaction of an anion such as a full secret compound such as bribe. It is also possible to prepare a metal = ^ potassium or lithium salt or an alkaline earth metal (for example, calcium) salt. Qucilide compound and erlotini series compounds such as Jiefeihu, erlotini or its salt can be formulated into medicine, sputum administration form is also oral or parenteral, and static 1 1 subcutaneous (4) Invest in an individual such as a patient or a sick animal. For example, 俾 = = Ming Qu Qu Li Li Bin compound and erlotini series of chemical agents (also = er, sexual, drug. Can be strong as a sexual dilution of the sword or assimilable edible carrier through the system sealed in hard The shell-shaped shell gelatin capsule can be compressed into the spine 93 200835507 or can be directly mixed with the patient's food. For oral treatment, the compound is combined with one or more excipients, and the edible lozenge and cheek ingot are used. , tablets, capsules, elixirs, suspensions, syrups, pastes, etc. These compositions and preparations must contain at least 0.1% active agent. The percentage of the composition and 5 preparations can of course be changed, conveniently It is in the range of from about 2% to about 60% by weight of the given unit dosage form. The active compound level of such therapeutically useful compositions is such that effective dosage levels are available. Lozenges, throat tablets, pills, capsules Etc. also contains the following ingredients: binders such as tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; disintegrating agents such as corn starch, horse starch, alginic acid, etc.; lubricants Such as stearin Magnesium sulphate; and sweeteners such as sucrose, fructose, lactose or aspartame; and flavoring agents such as menthol, wintergreen oil or cherry. When the unit dosage form is a capsule, in addition to the materials of the aforementioned categories, Capsules contain a liquid carrier such as a vegetable oil or polyethylene glycol. There may be a plurality of such forms as coatings, or otherwise modifying the solid shape of the solid unit dosage form. For example, lozenges, pills or capsules may be a coating, a wax, a shellac or a sugar, etc. The syrup or elixir may contain a compound of the invention, sucrose or fructose as a sweetener, methyl p-hydroxybenzoate and propyl p-hydroxybenzoate as a preservative, dye And a flavoring agent such as a cherry flavor or a willing taste. Any material used in the preparation of any unit dosage form must be pharmaceutically acceptable and substantially non-toxic in its amount used. Further, the compounds of the present invention can be incorporated into the continuation. Release preparations and devices. Qucilide compounds and erlotini series compounds such as gefitini, erlotini or their salts can also be administered by intravenous infusion or by injection via intravenous injection or abdomen 94 20 0835507 技。. The active agent or its salt solution can be interface activity (4) i can be combined with non-toxic preparations can also be prepared in glycerol, lysine polyethylene glycol, 5 10 15 20 two = material and oil preparation. These preparations contain a preservative for preventing the growth of microorganisms. The probiotics include a two-agent or aqueous dispersion or a sterile powder containing the active ingredient, which is suitable for use in the preparation of a solution or a reconstitution solution or The dispersion agent is sealed to the vesicles. In summary, the final dosage form is manufactured and stored: sterile, fluid stable. Liquid carrier, or dispersion medium including, for example, water, ethanol, polyol (eg glycerol = , Bu, liquid polyethylene, etc.), vegetable oil, non-toxic glycerin vinegar, and complex = field = compound. Suitable fluid properties can be formed, for example, by the formation of vesicles, in the case of sputum liquids, or by Make the face active and restrained. The microbial action can be obtained by various antibacterial agents and antifungal agents: for example, p-hydroxybenzoic acid esters, gas butanol, phenol, sorbic acid, Liunan = equal to the evening, preferably including an isotonic agent, for example Sugar, buffered with 2 sodium chloride. The composition for injection can be extended by the use of delayed absorption of, for example, aluminum stearate and gelatin in the composition. Once a sterile injectable solution is prepared by incorporating the compound of the present invention into the appropriate t solvent as needed, followed by extinction =. In the case of preparing a non-® powder for use in the preparation of the money injection solution, the method is a vacuum drying method and a cold 'east drying technique, and the active ingredient is added to obtain the desired component which is present in the aforementioned money-transitioned solution. Loose film J ο 95 200835507 For topical administration, the triclinide compound and the erlotini series of compounds such as gefitini, erlotini or its salt can be used in pure form, that is, in liquid form. However, it is generally desirable to combine a carrier acceptable for the skin, which may be a solid or a liquid, and which is applied to the skin as a composition or formulation. 5 Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, helium oxygen, aluminum oxide, and the like. Useful liquid carriers include water, alcohols or glycols or water-alcohol/glycol blends wherein the compound of the invention can be dissolved or dispersed in an effective amount by means of a non-toxic surfactant, if desired. Adjuvants such as perfumes and additional antimicrobial agents can be added to achieve the 10 best properties for a given application. The resulting liquid composition can be applied by an absorbent pad for impregnating bandages and other dressings, or by spraying a sprinkler or aerosol sprayer onto the affected area. Thickeners such as synthetic polymers, fatty acids, fatty acid hydrochloric acid, and fatty acid esters, fatty alcohols, modified celluloses, or modified mineral materials can also be used in liquid carriers to form expandable pastes, Gels, ointments, interest agents and the like are for direct application to the skin of the user. Examples of useful dermal compositions that can be used to deliver a compound of the invention to the skin are disclosed in Jacquet et al. (U.S. Patent No. 4,608,392), Geria (U.S. Patent No. 4,992,478), Smith et al. (U.S. Patent No. 4,559,157) &;w〇ltzman (US Patent No. 20 4,820,508). Useful doses of the pharmaceutical compositions of this invention can be determined by comparing their activity in vitro and in vivo activity in animal research models. Methods for extrapolating to humans for effective doses of small milk and other animals are known to the art; for example, see U.S. Patent 4,938,949. 96 200835507 In one non-limiting embodiment, the concentration of active agent in a liquid composition such as a lotion can range from about 0.1 to 25 wt.-% or from about 0.5 to 10 wt.-%. In a case of a shell, in a semi-solid composition or a solid composition such as a gel or a dispersion, the 'degree of time is about 1-5·1-5 wt.%, preferably about 0.5-2.5 wt·- %. In the case of a total of 5, the single dose of the main shot, the round or the food is usually 5-1500 * g, can be administered U times a day to obtain about 0.1-50 mg / kg / adult for adults The non-limiting dose of the present invention is 7.5 mg to 45 mg per ounce of oral administration, and the individual body weight can be appropriately adjusted.

10 1510 15

20 如此，本發明包括一種藥學組成物，包括曲西立濱化合物及_洛堤尼㈣化合物例如傑f堤尼、爾洛堤尼或其鹽與樂學上可接受之載劑之組合。適合經口、經局部或經，道外投樂之樂學組成物包括定量曲西立濱化合物及爾洛堤尼系列化合物例如傑f堤尼、爾洛堤尼或其鹽，該組成物組成本發明之較佳實施例。於本發明之上下文中f投予個體特別為人體之劑量為足夠於合_間框影響病人二治療反應。熟諳技藝人士瞭解該劑量將依據多項因素決定了包括動物情況、動物體重及癌症之嚴重程度及癌症階適當劑量為於腫瘤組織獲得已知可發揮期望反應:曲西立濱化合物及爾洛堤尼系列化合物例如傑費堤尼:爾洛堤尼或其鹽之濃度。較佳劑量為可獲得癌細胞生長之最大抑制而無無法管控之副作用之用量。他2 (或其藥學:可接受之鹽）之投予可為連續或以分開間隔投予，如技=界熟諳技巧人士之判定。衣’心可由所揭示之抑制癌細胞生長方法獲益之哺艽動斗 97 200835507 種包括但非限於靈長類諸如猿、黑獲猩、彳星彳星、人、猴；居家動物（例如寵物）諸如犬、貓、天竺鼠、倉鼠、越南大肚豬、兔、及雪貂；農場家畜諸如牛、水牛、野牛、馬、驢、豬、羊、及山羊，典型常見於動物園之珍奇異獸諸如熊、 5獅、虎、豹、象、海馬、犀牛、長頸鹿、玲羊、樹懶、燈羚、斑馬、牛羚、土撥鼠、無尾熊、袋鼠、負鼠、浣熊、貓熊、鬣狗、海豹、海獅、海象、水獺、瓶鼻海豚、海豚及鯨。病人」及「個體」等詞於此處互換使用，意圖包括人類哺乳動物及非人哺乳動物物種。同理，試管試驗之本 10發明方法可用於此種哺乳動物物種之細胞。给要使用本發明方法治療之病人可使用醫界專聿人士已知之標準技術識別。提供下列實例以供舉例說明但非限制性。 8.實例 15 實例1 :試管内篩檢細胞系及NCI分集集合：全部細胞系皆可購自ATCC或如鈾文ό兒明（Cheng J.Q·等人，致癌基因，1997 n p.2793-2801; West K.A.等人，今日抗藥性，2〇〇2 5: p.234-248; Satyamoorthy K.等人，癌症研究，2〇〇1 61: 20 ρ·7318·7324)。NCI結構分集集合為選自於約⑽，刪化合物 NCI藥物倉儲中之μ%種化合物存庫。有_此等分集集合化合物之選擇、結構式及活性之深度資料可參考Να發展治療計劃網址。紅過Akt轉形細胞生長抑制之篩檢^經八反丁】轉形之 200835507 NIH3T3細胞或經LXSN載體轉移感染之NIH3T3對照細胞 (Cheng J.Q.等人，致癌基因，1997. 14: ρ·2793-2801)係接種於96孔組織培養孔板。使用5μΜ NCI分集集合化合物處理後，細胞生長係使用細胞力價（CellTiter) 96 —種溶液細胞增 5 生套件組（普米嘉公司（Promega))檢測。於經AKT2-轉形之 NIH3T3細胞但未經LXSN轉移感染之NIH3T3細胞中抑制生長之化合物被視為Akt抑制劑候選者，接受進一步分析。試管内蛋白質激酶、細胞存活及細胞凋亡檢定分析：試管内激酶係如前述進行(例如參考Jiang K·等人，Mol Cell 10 Biol，2000· 20: ρ·139-148)。細胞存活係以MTS(普米嘉公司）檢定分析。細胞凋亡係以附著素（annexin) V檢測，如前文說明進行（Jiang K.等人，Mol Cell Biol，2000· 20: p.139-148)。重組Akt及PDK1係購自上態生技公司（Upstate Biotechnology Inc) 〇 15 結果Thus, the present invention encompasses a pharmaceutical composition comprising a combination of a triclinide compound and a derotinic (d) compound such as jieti, lodoti or a salt thereof and a pharmaceutically acceptable carrier. A music composition suitable for oral, topical or meridian, and including a quantitative triconide compound and a erlotini series compound such as jiedini, erlotini or a salt thereof, the composition of the composition Preferred embodiments of the invention. In the context of the present invention, the dose of the individual, particularly the human body, is sufficient to effect the patient's two therapeutic responses. Those skilled in the art understand that the dosage will be determined by a number of factors including the animal's condition, the animal's weight and the severity of the cancer, and the appropriate dose of the cancer. The known response to the tumor tissue is known to be expected: the triclinide compound and the erlotini The concentration of a series of compounds such as gefitini: erlotini or its salt. Preferred dosages are those which provide the greatest inhibition of cancer cell growth without the uncontrollable side effects. The administration of him 2 (or its pharmacy: acceptable salt) may be administered continuously or at separate intervals, as determined by skilled practitioners. The 'heart' can be benefited from the disclosed methods of inhibiting cancer cell growth. 200835507 includes but is not limited to primates such as cockroaches, black cockroaches, comet comets, humans, monkeys; home animals (such as pets) ) such as dogs, cats, guinea pigs, hamsters, Vietnamese pot-bellied pigs, rabbits, and ferrets; farm livestock such as cattle, buffalo, bison, horses, donkeys, pigs, sheep, and goats, typical of zoos such as exotic animals such as Bear, 5 lion, tiger, leopard, elephant, seahorse, rhino, giraffe, ling sheep, sloth, lamp antelope, zebra, wildebeest, groundhog, koala, kangaroo, possum, raccoon, panda, hyena, Seals, sea lions, walruses, otters, bottlenose dolphins, dolphins and whales. The terms "patient" and "individual" are used interchangeably herein and are intended to include both human mammalian and non-human mammalian species. Similarly, the test method of the invention can be applied to cells of such mammalian species. Patients who are to be treated using the methods of the present invention can be identified using standard techniques known to those skilled in the medical community. The following examples are provided by way of illustration and not limitation. 8. Example 15 Example 1: Intra-tube screening cell line and NCI diversity collection: All cell lines can be purchased from ATCC or as uranium wenming (Cheng JQ et al., Oncogene, 1997 n p. 2793-2801) West KA et al., Drug Resistance Today, 2〇〇2 5: p.234-248; Satyamoorthy K. et al., Cancer Research, 2〇〇1 61: 20 ρ·7318·7324). The NCI structure diversity set is a library of μ% of the compound selected from the (10), deleted compound NCI drug storage. There are _ such diversity sets. The depth of selection, structure and activity of the compounds can be found in the Να Development Treatment Program website. Screening for growth inhibition of red over Akt transfected cells ^ 反丁】 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 35 ) was inoculated into 96-well tissue culture well plates. After treatment with 5 μΜ NCI diversity pooled compounds, cell growth was detected using the CellTiter 96-solution solution cell kit (Promega). Compounds that inhibit growth in NIH3T3 cells transfected with AKT2-transformed NIH3T3 cells but not infected with LXSN were considered candidates for Akt inhibitors and were subjected to further analysis. In vitro assay of protein kinase, cell survival, and apoptosis assay: The in vitro kinase system was performed as described above (for example, refer to Jiang K. et al., Mol Cell 10 Biol, 2000 20: ρ. 139-148). Cell viability was analyzed by MTS (Pumijia) assay. Apoptosis was detected by annexin V as previously described (Jiang K. et al., Mol Cell Biol, 2000 20: p. 139-148). Recombinant Akt and PDK1 were purchased from Upstate Biotechnology Inc 〇 15 Results

Akt發訊徑路之小分子抑制劑API-2之識別於人類癌症檢測出Akt經常改變，摧毁Akt徑路，誘導細胞凋亡且抑制腫瘤生長（Jetzt A.等人，癌症研究， 2003·63:ρ·697-706,2003)。如此，Akt被視為新穎癌症治療Identification of the small molecule inhibitor API-2 of Akt signaling pathways in human cancers detects frequent changes in Akt, destroys Akt pathways, induces apoptosis and inhibits tumor growth (Jetzt A. et al., Cancer Research, 2003.63) : ρ·697-706, 2003). As such, Akt is considered a novel cancer treatment

2〇劑發展之具有吸引力之分子標靶。為了識別Akt之小分子抑制劑，得自NCI之1，992種化合物化學存庫(NCI分集集合)評估於經過AKT2-轉形但未經空白載體LXSN轉移感染之 NIH3T3細胞之抑制劑。重複實驗顯示32種化合物只能於經 AKT2-轉形細胞抑制生長。其中最強力之化合物API-2 (NCI 99 200835507 識別符：NSC 154020)可於50 nM濃度遏止細胞生長。第ία 圖顯不API-2也稱作為曲西立濱之化學結構式 (Schweinsberg P.D·等人，Biochem Pharmacol，1981.30： p.2521-2526)。API-2選擇性抑制AKT-2轉形細胞優於未經轉 5 形之親代細胞的事實，提醒發明人判定API-2是否為AKT2 激酶抑制劑。為了達成此項目的，AKT2於使用API-2處理後，以得自經AKT-2轉形之NIH3T3細胞之抗-AKT2抗體免疫沉澱。AKT2免疫沉澱使用抗磷酸-Akt抗體進行免疫墨點分析。如第1B圖所示，API-2於絲胺酸_309及絲胺酸-474(二 10 者為AKT2完全活化所需）顯著抑制AKT2磷酸化(Datta S.R. 等人，基因Dev，1999.13: ρ·2905·2927)。因三種Akt之同基因形共享高度同系列且類似的結構式，故評估API-2對其激酶活性的影響。HEK293細胞於EGF(50奈克/毫升)刺激前，以HA-Aktl、-AKT2及-AKT3處理，血清匱乏隔夜且以API-2 15 處理60分鐘。重複三次實驗顯示API_2遏止由EGF-誘導之激酶活性以及Aktl、AKT2及AKT3之磷酸化（第1C圖）。但重組株組成活性AKT2 (Myr-AKT2)之激酶活性於試管内激酶反應中不會受API-2抑制（第1D圖），提示API-2不會於試管内直接抑制Akt，API-2既非作為ATP競爭者，也非作為結合 20 至Akt之活性位置的酶基質競爭者。 API-2不會抑制已知之Akt上游活化劑：文獻中明確記載，Akt藉胞外刺激及胞内信號分子諸如活性Ras及Src透過 PI3K相依性方式來活化。因此，由把定Akt上游分子將可導致API-2抑制Akt。因PI3K及PDK1為Akt之直接上游調節劑 100 200835507 (Datta S.R·等人，基因Dev，1999· 13: ρ·2905-2927)，檢驗 API-2是否抑制pi3K及/或pDKl。HEK293細胞經過血清匱乏’然後以API-2或PI3K抑制劑、瓦特曼寧於EGF刺激前處理30分鐘。PI3K以抗-ρΙΙΟα抗體免疫沉澱。免疫沉澱物使 5用1"1-4々作為酶基質接受試管ΡΙ3Κ激酶檢定分析。如第2Α 圖所示，EGF-誘導之PI3K活性藉瓦特曼寧抑制但未受API-2 抑制。欲評估API-2對PDK1之效應，重組PDK1促進AKT2 多肽之蘇胺酸-309磷酸化之檢定分析係於含磷脂基肌糖醇之脂質囊存在下使用。如第2B圖所示，檢定分析藉對照組 10 PDK1抑制劑史妥洛史寶靈（staur〇Sp〇rine)強力抑制（ic5〇=5 nM)。相反地，於最高測試濃度(51μΜ)，απ-2只顯示21% 檢定分析之抑制。為了進一步評估API-2對PDK1活化之影響，於ΗΕΚ293細胞經過ΑΡΙ-2處理後，檢查於絲胺酸_241 之PDK1之自行填酸化程度，絲胺酸_241為自我磷酸化且對 15活性有關鍵重要性之一個殘基(Datta S.R·等人，基因Dev， 1999·13:ρ·2905-2927)。重複三次實驗顯示PDK1之磷酸化程度不受API-2抑制（第2B圖）。但PI3K抑制劑瓦特曼寧可抑制 EGF-刺激 PDK1 (第 2B圖）。 API-2 對透過PKC、PKA、SGK、STAT、JNK、p382 attractive molecular targets for the development of agents. To identify small molecule inhibitors of Akt, 1,992 compound chemical libraries (NCI diversity sets) from NCI were evaluated for inhibitors of NIH3T3 cells that were transfected with AKT2-transformed but not transfected with the blank vector LXSN. Repeat experiments showed that 32 compounds could only inhibit growth by AKT2-transformed cells. One of the most potent compounds, API-2 (NCI 99 200835507 identifier: NSC 154020), inhibits cell growth at a concentration of 50 nM. The ία map shows that API-2 is also referred to as the chemical structural formula of tromethamine (Schweinsberg P. D. et al., Biochem Pharmacol, 1981. 30: p. 2521-2526). The fact that API-2 selectively inhibits AKT-2 transfected cells over parental cells without transgenic forms suggests that the inventors determined whether API-2 is an AKT2 kinase inhibitor. To achieve this, AKT2 was immunoprecipitated with anti-AKT2 antibody from NIH3T3 cells transduced with AKT-2 after treatment with API-2. AKT2 immunoprecipitation was performed using an anti-phospho-Akt antibody for immunoblotting. As shown in Figure 1B, API-2 significantly inhibits AKT2 phosphorylation in serine _309 and serine-474 (two of which are required for complete activation of AKT2) (Datta SR et al., Gene Dev, 1999. 13: ρ · 2905·2927). Since the three Akt homologues share a highly homologous and similar structural formula, the effect of API-2 on its kinase activity was evaluated. HEK293 cells were treated with HA-Aktl, -AKT2 and -AKT3 prior to stimulation with EGF (50 Ng/ml), serum was depleted overnight and treated with API-2 15 for 60 minutes. Three experiments were repeated showing that API_2 inhibits EGF-induced kinase activity and phosphorylation of Aktl, AKT2 and AKT3 (Fig. 1C). However, the kinase activity of recombinant AKT2 (Myr-AKT2) is not inhibited by API-2 in the in vitro kinase reaction (Fig. 1D), suggesting that API-2 does not directly inhibit Akt in vitro, API-2 Not as an ATP competitor, nor as an enzyme matrix competitor that binds to the active site of 20 to Akt. API-2 does not inhibit known Akt upstream activators: it is clearly documented in the literature that Akt is activated by extracellular stimuli and intracellular signaling molecules such as active Ras and Src in a PI3K-dependent manner. Therefore, by targeting the upstream molecule of Akt, it will cause API-2 to inhibit Akt. Since PI3K and PDK1 are direct upstream regulators of Akt 100 200835507 (Datta S.R. et al., Gene Dev, 1999. 13: ρ·2905-2927), it was examined whether API-2 inhibits pi3K and/or pDKl. HEK293 cells were serum depleted and then treated with API-2 or PI3K inhibitor, Watmaning for 30 minutes prior to EGF stimulation. PI3K was immunoprecipitated with an anti-ρΙΙΟα antibody. The immunoprecipitate was assayed by using the 1"1-4々 as the enzyme substrate for the test tube ΡΙ3Κ kinase assay. As shown in Figure 2, EGF-induced PI3K activity was inhibited by Watmanin but not by API-2. To assess the effect of API-2 on PDK1, assays for recombinant PDK1 to promote phosphorylation of sulphate-309 phosphorylation of AKT2 polypeptide were used in the presence of phospholipid-containing myositol-containing lipid vesicles. As shown in Fig. 2B, the assay was strongly inhibited by the control group 10 PDK1 inhibitor staur〇Sp〇rine (ic5〇=5 nM). Conversely, at the highest test concentration (51 μΜ), απ-2 showed only 21% inhibition of the assay. In order to further evaluate the effect of API-2 on PDK1 activation, the ΗΕΚ293 cells were treated with ΑΡΙ-2, and the degree of self-priming of PDK1 of serine _241 was examined. Serine _241 was autophosphorylated and active for 15 A residue of critical importance (Datta SR et al., Gene Dev, 1999·13: ρ·2905-2927). Three experiments were repeated showing that the degree of phosphorylation of PDK1 was not inhibited by API-2 (Fig. 2B). However, the PI3K inhibitor Wattmannin inhibits EGF-stimulated PDK1 (Fig. 2B). API-2 pairs through PKC, PKA, SGK, STAT, JNK, p38

2〇及ERK發訊路徑之Akt有高度選擇性：Akt屬於AGC (PKA/PKG/PKC)激酶家族，該家族也包括PKA、PKC、血清可誘導激酶及糖皮質激素可誘導激酶(SGK)、P90核糖體 S6激酶、p70s6K、有絲***原-及應力-活化之蛋白質激酶及 PKC相關;敫酶。於AGCi敫酶家族中，PKA、PKC及SGK之 101 200835507 蛋白質結構式比其它成員更加接近Akt激酶之蛋白質結構式。因此，其次檢驗API-2對三種激酶之酶催化活性之影響。HEK293細胞以經ha-加標籤之PKA、PKCa或SGK轉移感染。试官内激酶檢定分析及免疫墨點分析顯示pKA及 5 PKC α之激酶活性分別受一種PKC抑制劑亦即PKAI及R〇 31-8220之抑制；而aPI_2對其活性不具影響（第2C圖及第2E 圖）。此外’血清可誘導之SGK激酶活性藉瓦特曼寧衰減但不藉API-2衰減（第2D圖）。此外，判定API_2是否對其它致癌基因存活徑路有影響。使用市售抗磷酸抗體之西方墨點 10 分析顯示Stat3、JNK、p38及Erkl/2之填酸化程度不受API-2 處理之影響（第2F圖）。此等資料指示API-2特異性抑制Akt 發訊徑路。於Akt過度表現之/活化之人癌細胞系中api-2遏止細跑生長且誘導細胞凋亡：API-2選擇性抑制Akt徑路之能 15 力’提示因偏好於有Akt之失序表現/活化之該等腫瘤細胞抑制增生及/或誘導細胞凋亡。因Akt過度表現或PTEN突變常見導致人惡性病中之Akt之活化，故API-2用來治療經由 AKT2過度表現（經〇VcAR3、OVCAR8、PANC1 及AKT2-轉形之NIH3T3)或PTEN基因突變（PC-3、LNCaP、 20 MDA_MB-468)引發表現組成活性Akt之細胞、以及不會表現之細胞（OVCAR5、DU-145、T47D、C0L0357 及 LXSN-NIH3T3)、以及由IGF-1活化來活化Akt或對藉IGF-1 之生長刺激不會反應之黑素瘤細胞（Satyamoorthy K.等人，癌症研究，2001. 61: ρ·7318-7324)。免疫墨點分析顯示只有 102 200835507 於表現升高之Akt或對IGF-1刺激有反應之細胞中，Akt之磷酸化程度才受API-2抑制（第3A圖）。如此，API-2於Akt過度表現/活化細胞中抑制細胞生長達比較含低含量Akt之細胞遠更高程度。如第3B圖所示，於Akt過度表現/活化細胞系、 5 LNCaP、PC-3、OVCAR3、OVCAR8、PANC1 、 MDA-MB-468、及WM35中，API-2處理抑制細胞增生達約 50-60% ;於DU145、OVCAR5、C0L0357、T47D及WM852 細胞只抑制細胞增生達約10_20%，該等細胞含低濃度Akt，或對藉IGF-1之生長刺激無反應。此外，API-2誘導細胞凋 10 亡達 8倍（OVCAR3)、6倍（OVCAR8)、6倍(PANC1)、及3倍 (AKT2-NIH3T3)。於OVCAR5、C0L0357及LXSN-NIH3T3 細胞中，API-2處理與載媒劑(DMSO)處理間並未觀察得細胞凋亡有顯著差異。如此，於表現失序Akt中，API-2偏好於表現失序Akt之細胞中抑制細胞生長及誘導細胞凋亡。 15 API-2抑制Akt下游標靶：業已顯示Akt透過多種蛋白質之磷酸化來發揮其細胞效果（Datta S_R.等人，基因Dev， 1999·13··ρ·2905-2927)。已經識別多於20種蛋白質為Akt酶基質，包括插子頭蛋白質家族成員（FKHR、AFX及FKHRL1)、結節素（tuberlin)/TSC2、p70S6K、GSK-3 /3、p21WAF1/cipl、 20 p27kipl、MDM2、Bad、ASK1 及ΙΚΚα 等。其次檢驗API-2 是否抑制Akt下游標靶。因抗-磷酸_結節素、_Bad、_AFX、及-GSK-3/5抗體為市面上可得，因此判定api_2對於其藉 Akt誘導磷酸化之效果。於API-2 (ΙμΜ)處理後，OVCAR3 細胞經溶解且使用個別抗磷酸抗體進行免疫墨點分析。第 103 200835507 4A圖顯示AI>I-2顯著抑制結節素之填酸化程度，結果導致結節素之穩定與向上調節（Dan，H c，、，jBi〇lChem，2002· 277: ρ·35364-35370)。Bad、GSK-3)S、及AFX之磷酸化程度部分受API-2衰減。此等資料提示API-2經由抑制其下游 5標靶之磷酸化來誘導細胞死亡及細胞生長停止。API-2以不等程度抑制Akt下游標靶可能原因係在於此等標靶之磷酸化位置也受其它激酶的調節，例如Bad絲胺酸-136除了藉 Akt碟酸化之外也藉PAKU粦酸化（Schurmann，A.等人，Mol Cell Biol，2000· 20: ρ·453-461)。 l〇實例2 :於裸小鼠腫瘤異種移植研究模型中之抗腫瘤活性腫瘤細胞經收穫，懸浮於PBS，且如先前報告皮下注射於8週齡雌裸小鼠之右與左脅腹(2χ1〇6細胞/脅腹）（sun j. 等人，癌症研究，1999· 59: ρ·4919-4926, 1999)。當腫瘤達到約100-150立方毫米時，動物經隨機分配，經每日腹内注 15 射0·2毫升曲西立濱化合物及/或爾洛堤尼系列化合物例如傑費堤尼、爾洛堤尼或其鹽之載媒劑。對照動物接受 DMSO(20%)載媒劑，而處理組動物注射ΑΠ-2 (1毫克/千克/ 日）於20% DMSO。 API-2於過度表現Akt之裸小鼠抑制腫瘤之生長：於人Akt and ARK signaling pathways are highly selective: Akt belongs to the AGC (PKA/PKG/PKC) kinase family, which also includes PKA, PKC, serum-inducible kinases, and glucocorticoid-inducible kinase (SGK), P90 ribosomal S6 kinase, p70s6K, mitogen- and stress-activated protein kinases and PKC-related; chymase. In the AGCi chymase family, the structural formula of PKA, PKC and SGK 101 200835507 is closer to the protein structure of Akt kinase than other members. Therefore, the effect of API-2 on the enzymatic activity of the three kinases was examined. HEK293 cells were transfected with ha-tagged PKA, PKCa or SGK. The assay of the kinase assay and the immunoblot analysis showed that the kinase activities of pKA and 5 PKC α were inhibited by a PKC inhibitor, namely PKAI and R〇31-8220, respectively; while aPI_2 had no effect on its activity (Fig. 2C and Figure 2E). In addition, serum-inducible SGK kinase activity was attenuated by Watmanin but not by API-2 (Fig. 2D). In addition, it was determined whether API_2 had an effect on the survival pathway of other oncogenes. Western blots using commercially available anti-phospho-antibody 10 analysis showed that the degree of acidification of Stat3, JNK, p38 and Erkl/2 was not affected by API-2 treatment (Fig. 2F). These data indicate that API-2 specifically inhibits the Akt signaling pathway. In the overexpressed/activated human cancer cell line of Akt, api-2 inhibits fine-grain growth and induces apoptosis: API-2 selectively inhibits the ability of the Akt pathway 15 stress' suggesting a preference for Akt's disordered performance/ The activated tumor cells inhibit proliferation and/or induce apoptosis. API-2 is used to treat overexpression of AKT2 (via VcAR3, OVCAR8, PANC1 and AKT2-transformed NIH3T3) or PTEN gene mutations due to Akt overexpression or PTEN mutations leading to activation of Akt in human malignancies ( PC-3, LNCaP, 20 MDA_MB-468) prime cells expressing active Akt, as well as cells that are not expressed (OVCAR5, DU-145, T47D, C0L0357, and LXSN-NIH3T3), and activation of Akt by activation of IGF-1 Or melanoma cells that do not respond to growth stimulation by IGF-1 (Satyamoorthy K. et al., Cancer Research, 2001. 61: ρ·7318-7324). Immunoblot analysis showed that only the degree of phosphorylation of Akt was inhibited by API-2 in cells with elevated Akt or response to IGF-1 stimulation in 102 200835507 (Fig. 3A). Thus, API-2 inhibits cell growth in Akt overexpressing/activating cells to a much higher degree than cells containing low levels of Akt. As shown in Figure 3B, in the Akt overexpressing/activating cell line, 5 LNCaP, PC-3, OVCAR3, OVCAR8, PANC1, MDA-MB-468, and WM35, API-2 treatment inhibited cell proliferation by approximately 50- 60%; cells in DU145, OVCAR5, C0L0357, T47D and WM852 inhibited cell proliferation by only about 10-20%, and these cells contained low concentrations of Akt, or did not respond to growth stimulation by IGF-1. In addition, API-2 induced cell death by 8 fold (OVCAR3), 6 fold (OVCAR8), 6 fold (PANC1), and 3 fold (AKT2-NIH3T3). In OVCAR5, C0L0357 and LXSN-NIH3T3 cells, no significant differences in apoptosis were observed between API-2 treatment and vehicle (DMSO) treatment. Thus, in the performance of disordered Akt, API-2 prefers to inhibit cell growth and induce apoptosis in cells exhibiting disordered Akt. 15 API-2 Inhibits Akt Downstream Targets: Akt has been shown to exert its cellular effects through phosphorylation of various proteins (Datta S_R. et al., Gene Dev, 1999.13··ρ 2905-2927). More than 20 proteins have been identified as Akt enzyme substrates, including insert protein family members (FKHR, AFX and FKHRL1), tuberlin/TSC2, p70S6K, GSK-3/3, p21WAF1/cipl, 20 p27kipl, MDM2, Bad, ASK1, and ΙΚΚα. Secondly, it was tested whether API-2 inhibits Akt downstream targets. Since anti-phospho- nodulin, _Bad, _AFX, and -GSK-3/5 antibodies are commercially available, the effect of api_2 on phosphorylation by Akt was determined. After treatment with API-2 (ΙμΜ), OVCAR3 cells were lysed and immunoblot analysis was performed using individual anti-phospho antibodies. Section 103 200835507 4A shows that AI>I-2 significantly inhibits the degree of acidification of nodule, resulting in the stability and upregulation of nodulin (Dan, H c,,, jBi〇lChem, 2002· 277: ρ·35364-35370) ). The degree of phosphorylation of Bad, GSK-3)S, and AFX is partially attenuated by API-2. These data suggest that API-2 induces cell death and cell growth arrest by inhibiting phosphorylation of its downstream 5 targets. The possible reason for API-2 to inhibit Akt downstream targets in varying degrees is that the phosphorylation sites of these targets are also regulated by other kinases. For example, Bad serine-136 is acidified by PAKU in addition to acidification by Akt. (Schurmann, A. et al., Mol Cell Biol, 2000·20: ρ·453-461). Example 2: Antitumor activity in a nude mouse tumor xenograft study model Tumor cells were harvested, suspended in PBS, and injected subcutaneously into the right and left flank of 8 week old female nude mice as previously reported (2χ1) 〇6 cells/flank (sun j. et al., Cancer Research, 1999. 59: ρ·4919-4926, 1999). When the tumor reaches about 100-150 cubic millimeters, the animals are randomly assigned, and a daily intraperitoneal injection of 0.2 ml of troxazide compound and / or erlotini series of compounds such as Gefitini, Erlo A carrier agent for Tenny or its salt. Control animals received DMSO (20%) vehicle, while treatment animals injected ΑΠ-2 (1 mg/kg/day) in 20% DMSO. API-2 inhibits tumor growth in nude mice overexpressing Akt: in humans

2〇卵巢癌及胰癌中顯示AKT1及AKT2之經常性過度表現/活化及/或擴增（Cheng J.Q·及Nicosia S.V·，於癌症百科參考， 2001，Schwab D·，編輯Springer，柏林、海德堡及紐約，35_37 頁）。Akt徑路受PI3K、HSP70、Src及法尼基轉移酶等抑制劑的抑制，導致細胞生長停止與細胞凋亡的誘導(S〇litD.B 104 200835507 等人，癌症研究，2003· 63: ρ·2139-2144; XU，W.等人，癌症研究，2003· 63: ρ·7777-7784)。晚近研究顯示Akt升高之異種移植之腫瘤生長也受腫瘤内注射優勢陰性Akt腺病毒的顯著抑制（Jetzt A.等人，癌症研究，2003. 63: p.697-706, 5 2003)。因ΑΠ-2抑制Akt發訊，且只於Akt濃度升高之癌細胞誘導細胞凋亡及細胞生長停止（第3圖），故有升高之Akt 含量之腫瘤生長應比較裸小鼠體内有較低含量Akt之腫瘤之腫瘤生長，對API-2更敏感。為了達到此項目的，皮下注射Akt過度表現細胞（〇vCAR3、OVCAR8及PANC-1)皮下植 10 入右脅腹，而表現低度Akt之該等細胞系（OVCAR5及 C0L0357)則植入小鼠的左脅腹。當腫瘤達到平均大小約為 100-150立方毫米時，動物隨機分組且使用載媒劑或api-2 (1毫克/千克/日）經腹内注射處理。如第4B圖所示，以載媒劑處理之0 VC AR5及C0L03 57腫瘤於腫瘤植入後49曰生長 15 至約800-1000立方毫米。以載媒劑對照組處理之OVCAR3、 OVCAR8及PANC-1腫瘤，於植入後49日生長至約700-900 立方毫米。API_2抑制OVCAR3、OVCAR8及PANC-1腫瘤生長分別達90%、88%及80%。相反地，API-2對裸小鼠之 OVCAR5及C0L0357之細胞生長極少有影響（第4B_4D圖而 20資料未顯示）。於1毫克/千克/日劑量，API-2對小鼠血糖濃度、體重、活動性及食物的攝取無影響。於接受處理之腫瘤樣本中’ Akt活性受API-2影響，但總Akt含量不變（第4E 圖）。共同考量，此等結果指示API-2選擇性抑制有升高之 Akt濃度之腫瘤的生長。 105 200835507 實例3 : TCN直接抑制野生型Akt激酶活性 API-2 (TCN)於試管内直接抑制由PDK1所誘導之野生型Akt激酶活性（第1圖）。此等結果證實API-2為直接Akt抑制劑，潛在機轉可能為API-2結合至Akt之PH功能部位及/或蘇 5 胺酸-308。於含有磷脂基肌糖醇-3,4,5-P3 (PIP3)、API-2及組織腺H2B作為酶基質之激酶緩衝液中，以PDK1與Akt之重組株進行試管内激酶檢定分析。培養30分鐘後，藉 SDS-PAGE分離反應且暴露於薄膜。實例4 : TCN於癌症抗性細胞中有效 10 TCN(API-2)之效果係於西鉑汀、太平洋紫杉醇、及塔莫西芬抗藥性A270CP、C_13、OVCAR433及MCF7/TAM細胞中試驗。於此等細胞中，API-2克服西鉑汀、太平洋紫杉醇、及塔莫西芬抗藥性。實例5: TCN增強由崔茲竹美所抑制之生長且誘導細胞凋亡 15 材料及方法細胞系及細胞培養：腫瘤發生BT474.ml亞系維持於杜別可（DulbeCC〇，s)改性鷹氏（Eagle’s)培養基：漢氏（Ham，s) F-12培養基（1:1)含8-l〇%FBS。抗體及試劑：崔茲竹美由基因科技公司（Genentech)(加 20州舊金山）贈與。RAD〇〇l(艾福羅里幕斯（ever〇iimus))由諾華公司（Novartis)(紐澤西州東漢諾瓦）贈與，qlt〇267及 KP372-1係由QLT公司（BC溫哥華）贈與。曲西立濱（6_胺基 -4-甲基-8-(3-D-核糖呋喃糖基)-4H，8H-吡咯并[4,3,2-de]嘧啶[4,5-c]嗒畊）係購自拜瑞關係企業（Berry & Ass(>dates， 106 2008355072〇 Ovarian cancer and pancreatic cancer show frequent overexpression/activation and/or expansion of AKT1 and AKT2 (Cheng JQ· and Nicosia SV·, in Cancer Encyclopedia Reference, 2001, Schwab D·, Editing Springer, Berlin, Heidelberg And New York, page 35_37). The Akt pathway is inhibited by inhibitors such as PI3K, HSP70, Src, and farnesyltransferase, leading to cell growth arrest and induction of apoptosis (S〇litD.B 104 200835507 et al., Cancer Research, 2003. 63: ρ · 2139-2144; XU, W. et al., Cancer Research, 2003· 63: ρ·7777-7784). Recent studies have shown that tumor growth in xenografts with elevated Akt is also significantly inhibited by intratumoral injection of dominant negative Akt adenovirus (Jetzt A. et al., Cancer Research, 2003. 63: p. 697-706, 5 2003). Because ΑΠ-2 inhibits Akt signaling and induces apoptosis and cell growth only in cancer cells with elevated Akt concentrations (Fig. 3), tumor growth with elevated Akt levels should be compared to nude mice. Tumor growth in tumors with lower Akt levels is more sensitive to API-2. In order to achieve this, subcutaneous injection of Akt overexpressing cells (〇vCAR3, OVCAR8, and PANC-1) subcutaneously implanted into the right flank, while those cells exhibiting low Akt (OVCAR5 and C0L0357) were implanted in mice. The left flank. When the tumors reached an average size of about 100-150 mm3, the animals were randomized and treated with vehicle or api-2 (1 mg/kg/day) by intraperitoneal injection. As shown in Figure 4B, the vehicle-treated 0 VC AR5 and C0L03 57 tumors grew 15 to about 800-1000 mm 3 at 49 肿瘤 after tumor implantation. The OVCAR3, OVCAR8, and PANC-1 tumors treated with the vehicle control group grew to about 700-900 cubic millimeters 49 days after implantation. API_2 inhibited OVCAR3, OVCAR8 and PANC-1 tumor growth by 90%, 88% and 80%, respectively. In contrast, API-2 had little effect on cell growth of OVCAR5 and C0L0357 in nude mice (Fig. 4B_4D and 20 data not shown). At 1 mg/kg/day, API-2 had no effect on blood glucose concentration, body weight, activity and food intake in mice. The Akt activity was affected by API-2 in the treated tumor samples, but the total Akt content was unchanged (Fig. 4E). Taken together, these results indicate that API-2 selectively inhibits the growth of tumors with elevated Akt concentrations. 105 200835507 Example 3: TCN directly inhibits wild-type Akt kinase activity API-2 (TCN) directly inhibits wild-type Akt kinase activity induced by PDK1 in vitro (Fig. 1). These results confirm that API-2 is a direct Akt inhibitor and that the potential mechanism may be the binding of API-2 to the PH functional site of Akt and/or sulphate-308. Intracellular kinase assay was performed using a recombinant strain of PDK1 and Akt in a kinase buffer containing phospholipid-based myositol-3,4,5-P3 (PIP3), API-2 and histidine H2B as an enzyme substrate. After 30 minutes of incubation, the reaction was separated by SDS-PAGE and exposed to the membrane. Example 4: TCN is effective in cancer-resistant cells The effect of 10 TCN (API-2) was tested in citrate, paclitaxel, and tamoxifen-resistant A270CP, C_13, OVCAR433, and MCF7/TAM cells. In these cells, API-2 overcomes the resistance of cetamine, paclitaxel, and tamoxetine. Example 5: TCN enhances growth inhibited by Cui Zhumei and induces apoptosis 15 Materials and methods Cell lines and cell culture: Tumor-generating BT474.ml sub-lines maintained in Dulbec (s) modified eagle Eagle's medium: Hans (Ham, s) F-12 medium (1:1) containing 8-l〇% FBS. Antibodies and Reagents: Cui Zhemei was donated by Genentech (plus San Francisco, 20). RAD〇〇l (ever〇iimus) was donated by Novartis (East Hanover, New Jersey), and qlt〇267 and KP372-1 were donated by QLT (BC Vancouver) . Trifluralin (6-amino-4-methyl-8-(3-D-ribosefuranosyl)-4H,8H-pyrrolo[4,3,2-de]pyrimidine [4,5-c ]嗒耕) is purchased from Berry & Ass (>dates, 106 200835507

Inc·)(密西根州安哈博）。艾朵弗新(Edelfosine)係購自卡爾生化公司（Calbiochem)(加州聖地牙哥）。合成選擇性Akt抑制劑 4ADPIB(4-胺基-2)-(3,4-二氯-苯基)-Ν-(1Η-吲唑 5-基)-丁醯胺）（美國專利6,919,340)。PTEN抗體係得自聖塔克魯 5 兹生技公司（Santa Cruz Biotechnology)(加州聖塔克魯茲）。3-肌動蛋白抗體係得自西革瑪公司（Sigma)(密蘇里州聖路易）。全部其它抗體皆係購自細胞發訊技術公司（Cell Signaling Technology)(麻省丹佛）。 PTEN反訊息及非特異性募核苔酸一過性轉移感染：對 10 PTEN特異性反訊息(AS)募核苷酸、對照組非特異性(NS)寡核苷酸及轉移感染程序係遵照Nagata Y·等人，癌細胞，2〇〇4, 6(2):ρ·117-27 進行。細胞增生檢定分析··經PTEN AS/NS轉移感染之 BT474.ml細胞接種成2500細胞/0.32平方厘米孔。細胞單獨 15 以Akt/mTOR徑路抑制劑或組合崔兹竹美如所述處理5曰，使用細胞力價96AQ非放射性細胞增生檢定分析套件組，根據製造商方案（普米嘉公司，威斯康辛州麥迪遜），藉MTS 檢定分析測定存活細胞。處理後之細胞與對照組經DMSO 處理之BT474.ml細胞比較，計算生長抑制百分比。 20 APO-BRDU TUNEL檢定分析：經PTEN AS及NS轉移感染之BT474.ml細胞接種於6孔孔板(4-6xl05細胞/孔）。於接種後24小時，細胞如指示以崔茲竹美、TCN及/或RAD001 處理72小時。使用APO-BRDU TUNEL檢定分析套件組(鳳凰流動系統公司（Phoenix Flow Systems)，加州聖地牙哥）， 107 200835507 根據製造商方案，收集漂浮細胞及黏著細胞，加標記及染色。使用FAC掃描（FACScan)流動細胞計及CellQuest Pro 4·〇2軟體（貝克頓迪根森公司（Becton Dickinson)，紐澤西州法蘭克林湖）收集與分析資料。至少檢驗1〇,〇〇〇事件。 SDS-PAGE及免疫墨點分析：以PTENAS/NS寡核苷酸轉移感染之細胞係如指示處理。免疫墨點係如所述藉Inc.) (Anhalb, Michigan). Edelfosine was purchased from Calbiochem (San Diego, Calif.). Synthetic Selective Akt Inhibitor 4ADPIB(4-Amino-2)-(3,4-dichloro-phenyl)-indole-(1Η-indazol-5-yl)-butanamine) (U.S. Patent 6,919,340). The PTEN anti-system was obtained from Santa Cruz Biotechnology (Santa Cruz, Calif.). The 3-actin resistance system was obtained from Sigma (St. Louis, Missouri). All other antibodies were purchased from Cell Signaling Technology (Denver, MA). PTEN anti-message and non-specific nucleotide transient transfer infection: compliance with 10 PTEN-specific anti-message (AS) nucleotides, control non-specific (NS) oligonucleotides, and metastatic infection programs Nagata Y. et al., cancer cells, 2〇〇4, 6(2): ρ·117-27. Cell proliferation assay analysis BT474.ml cells infected with PTEN AS/NS were seeded into 2500 cells/0.32 cm2 wells. Cells alone 15 were treated with an Akt/mTOR pathway inhibitor or a combination of Cui Zhemei as described, using a cell-powered 96AQ non-radioactive cell proliferation assay kit, according to the manufacturer's protocol (Pumijia, Wisconsin, McGrady) (), by MTS assay analysis to determine viable cells. The percentage of growth inhibition was calculated by comparing the treated cells with the DMSO-treated BT474.ml cells of the control group. 20 APO-BRDU TUNEL assay: BT474.ml cells infected with PTEN AS and NS were seeded in 6-well plates (4-6 x 105 cells/well). At 24 hours post-inoculation, cells were treated with Triz, TCN and/or RAD001 for 72 hours as indicated. Use the APO-BRDU TUNEL assay suite (Phoenix Flow Systems, San Diego, CA), 107 200835507 Collect floating cells and adherent cells according to the manufacturer's protocol, labeling and staining. Data were collected and analyzed using the FAC Scan (FACScan) flow cytometer and the CellQuest Pro 4·〇2 software (Becton Dickinson, Franklin Lake, New Jersey). At least 1 检验, 〇〇〇 event. SDS-PAGE and immunoblot analysis: Cell lines infected with PTENAS/NS oligonucleotides were treated as indicated. Immune ink dots are as described

NagataY.等人，癌細胞，2〇〇4,6(2)φ·117-27進行。結果於ΡΤΕΝ缺陷細胞曲西立濱及RAD001增強由崔茲竹美Nagata Y. et al., cancer cells, 2〇〇4,6(2)φ·117-27. Results ΡΤΕΝ ΡΤΕΝ defective cell 曲西立滨 and RAD001 enhanced by 崔兹竹美

10之生長抑制：欲尋找克服崔茲竹美抗藥性之策略，特別因 ΡΤΕΝ損失所引起的抗藥性，試驗六種不同小分子抑制劑，其直接或間接靶定PI3K/Akt/mTOR信號轉導徑路，此徑路係由ErbB2過度表現及ΡΤΕΝ損失所活化的主要徑路。目標係識別具有與崔兹竹美協同效應之化合物較佳於低劑量化 15合物來減低毒性。選用之藥物係靶定於Akt、mTOR及接合素鍵聯激酶(ILK)(第9A圖）。BT474.ml細胞為BT474乳癌細胞系之腫瘤發生亞系，且表現高濃度ErbB2。當ΡΤΕΝ濃度係經由以ΡΤΕΝ反訊息募核苷酸(PTEN AS)轉移感染而降低時，BT474.ml乳癌細胞變成比有正常PTEN濃度之細胞對崔 20兹竹美之抗增生效應更具有抗藥性，提供良好乳癌之實驗研究模型，其中崔茲竹美抗藥性係由於ΡΤΕΝ損失所引起 (Nagata Υ·等人，癌細胞，2004, 6(2): ρ·117-27及第 9Β及9C 圖）。非特異性寡核苷酸(NS)經轉移感染作為對照組。以 PTEN AS寡核苷酸處理可有效降低pTEN濃度（第11A及11B 108 200835507 白無法改變細胞的ErbB2 圖）。PTEN AS或NS對照募核誓酸含量。經PTENAS及聰⑽轉移感染之B觀如以六種化合物或«竹美單獨處理及組合處理5日，評估細胞增生且與經過DMSQ處理之對照組作比較。使用生長抑制作為生物端點，發明人比較各種藥物具有與崔兹竹美之協力效Growth inhibition of 10: To find a strategy to overcome the resistance of Cui Zhe Bamboo, especially the drug resistance caused by the loss of ΡΤΕΝ, test six different small molecule inhibitors, which directly or indirectly target PI3K/Akt/mTOR signal transduction Path, which is the main path activated by ErbB2 overexpression and enthalpy loss. The target identifies a compound having a synergistic effect with Tries, preferably a low dose compound to reduce toxicity. The drugs selected are targeted at Akt, mTOR and zygoxin-linked kinase (ILK) (Fig. 9A). The BT474.ml cell is a tumorigenic subline of the BT474 breast cancer cell line and exhibits a high concentration of ErbB2. BT474.ml breast cancer cells become more resistant to anti-proliferative effects of Cui 20zzhumei than cells with normal PTEN concentration when the concentration of sputum is reduced by transfection with sputum-reporting nucleotides (PTEN AS). Provides an experimental model for the study of good breast cancer, in which the Cui Zhemei resistance is caused by loss of sputum (Nagata Υ· et al, cancer cells, 2004, 6(2): ρ·117-27 and Β9 and 9C ). Non-specific oligonucleotides (NS) were metastatically infected as a control group. Treatment with PTEN AS oligonucleotides effectively reduced the concentration of pTEN (11B and 11B 108 200835507 white could not alter the ErbB2 map of cells). PTEN AS or NS control raised nuclear acid content. B-views of PTENAS and Cong (10) metastatic infections were evaluated by six compounds or with Zhumei alone and in combination for 5 days, and cell proliferation was evaluated and compared with the DMSQ-treated control group. Using growth inhibition as a bioend point, the inventors compared various drugs to have synergy with Tries.

果，使用單職藥時可獲得_2(MG%生長㈣之藥物（第9A 圖）〇If you use a single-agent, you can get _2 (MG% growth (4) drug (Figure 9A)〇

幾乎全部化合物皆顯示生長抑制效果特別於高濃度且 10於有完MpTEN之細胞顯示生長抑制效果（第9八圖）。但其中兩種化合物亦即曲西立濱及RAD〇〇1當與崔兹竹美組^時比較崔级竹美或任一種化合物單獨投藥時，組合投藥可顯著提升於PTEN AS細胞之生長抑制（第9八圖）。曲西立濱（也稱作為API-2)為可抑制Akt活化之化合物，增強由崔茲竹美 15產生之生長抑制於20倍濃度範圍（第9B圖）。當以低劑量(<ι nM)投予RAD001時，mTOR抑制劑RAD〇〇1(艾福羅里幕斯) 可提高由崔兹竹美產生之生長抑制（第9C圖）。令人訝異地，曲西立濱及RAD001可與崔茲竹美協力來於PTENAS及 PTEN NS細胞以類似程度抑制細胞生長（第9B圖及第9C 20 圖）。要言之，曲西立濱及RAD001可回復PTEN缺陷細胞之崔茲竹美敏感度。曲西立濱及RAD001也可於PTENAS細胞及PTEN NS細胞二者有效作為單一藥劑，對曲西立濱及 RAD001分別係大於5 nM及1.5 nM (第9B圖及第9C圖）。第三化合物亦即ILK抑制劑QLT0267可於狹窄劑量範 109 200835507 圍（〜5-15 pM)增強由崔茲竹美產生的生長抑制（第9A圖）。因 QLT0267與崔茲竹美具有協力效果之劑量範圍狹窄，故未進一步研究此種化合物。於大於2〇pM之濃度，QLT0267與崔茲竹美不具協力效果，反而呈單一作用劑顯著抑制細胞 5 生長。於組合治療後之細胞凋亡之誘導：為了評估生長抑制是否伴隨細胞凋亡，發明人單獨與曲西立濱、RAD001及崔茲竹美或組合使用來處理經PTEN AS及NS轉移感染之 BT474.ml細胞，且定量細胞凋亡程度（第1〇圖）。RAD001單 10 獨或與崔茲竹美組合並未顯著誘導細胞凋亡。雖然於單獨使用崔茲竹美或曲西立濱處理後，TUNEL-陽性細胞凋亡細胞數目微增，但此種增加不具有統計意義。但比較於經 PTEN AS及NS轉移感染細胞二者中之全部其它處理，曲西立濱與崔茲竹美之組合物可顯著誘導細胞凋亡（第10圖）。 15 實例6 : TCN抑制下游發訊分子之Akt及mTOR抑制之活化：免疫墨點分析證實曲西立濱及RAD001可阻斷定Akt 及mTOR (藉ErbB2活化之重要發訊分子）及曲西立濱及 RAD001之標靶之活化。Akt用於Thr308及Ser473之磷酸化經分析作為Akt活性之指標；mTOR活性係藉p70S6K 20 (70-kDa核糖體蛋白質S6激酶）亦即mTOR標靶之磷酸化評估。於曲西立濱處理後，於兩個位置之Akt磷酸化實質上減低（第11A圖）。於PTEN-缺陷細胞，於曲西立濱與崔茲竹美組合處理後，Akt磷酸化程度係類似於有完好pteN之細胞中所見（第11A圖’線道4及8)。如此，經由有效阻斷Akt活 110 200835507 化，曲西立濱克服PTEN損失之不良影響。RAD001可顯著阻斷P70S6K之磷酸化（第11B圖）。但RAD001與崔茲竹美的組合不會降低P70S6K磷酸化超過單獨使用RAD001所見（第 11B圖）。晚近識別回授回路，其可導致於使用mTOR抑制劑 5 諸如RAD001處理後，Akt之磷酸化及活化(O’Reilly K.E.等人，癌症研究，2006· 66(3): P.1500-8)。也觀察到藉RAD001 以及藉崔茲竹美與RAD001之組合治療回授活化Akt，可消除藉此回授回路之Akt磷酸化（第11B圖，線道3相對於線道 4)，與於mTOR抑制後之Akt活化仰賴上游受體酪胺酸激酶 10 符合一致（〇，Reilly K.E.等人，癌症研究，2006. 66(3): P.1500-8)。要言之，藥物抑制其經預測之標靶激酶，組合治療比較任何單一藥劑對Akt/mTOR發訊徑路有更大抑制效果’即使於PTEN缺陷細胞亦如此。實例7 :於PTEN缺陷腫瘤TCN及崔茲竹美抑制腫瘤生長 15 材料及方法於SCID小鼠異種移植之人類腫瘤研究模型：雌性，6_ 週齡，嚴重組合免疫缺陷（SCID)小鼠係得自塔可尼(Tac〇nic) 農場（紐約州哈德遜）。腫瘤異種移植係如NagataY•等人，癌細胞，2004, 6(2): ρ·117-27所述進行。當異種移植腫瘤達到 2〇 100-150立方毫米平均大小時，小鼠分6組，各組有7頭小鼠及平均分布的腫瘤大小，且如後文說明處理。每週透過腫瘤内注射將PTEN反訊息（30 pg)募核苔酸投予各小鼠。於 PTEN AS募核苗酸投予後一週開始藥物的處理。崔兹竹美以每週兩次G·5毫克/千克劑量於皮升食齡經由多個部 111 200835507 位的腫瘤内注射來投予。崔茲竹美係以0.5毫克/千克/日於 200皮升20% DMSO食鹽水溶液劑量經腹内（ι·ρ·)注射投予。RAD001係透過胃管灌食以1皮克/千克於500皮升5%葡萄糖水劑量每週兩次投予。20% DMSO食鹽水溶液(200皮升 5 /日）經腹内注射投予。使用測微計每週兩次測量腫瘤尺寸，腫瘤體積計算為體積=長X寬2/2。統計分析：使用 GraphPad Prism 3·〇 for Windows (圖墊軟體公司（Graph Pad Software)，加州聖地牙哥）進行單向 ANOVA。 10 結果早期生物資料及分子資料極為具有展望，但活體内研究提供治療效果之最苛刻試驗。因此曲西立濱及RAD〇〇1 係於活體内試驗。BT474.ml細胞異種移植片注射入6週齡 SCID小鼠之***脂肪墊内部。於腫瘤形成後，小鼠每週經 15腫瘤内’主射接受PTEN AS。此方案於活體内可有效作為 PTEN缺陷腫瘤之模型（Nagata γ·等人，癌細胞，2_， 6(2):Ρ· 117-27)。小鼠隨機分組成為單獨或組合接受曲西立濱、RAD00卜崔茲竹美或DMS0之多個處理組。於處理後，單獨以DMSO、崔茲竹美、尺八：〇001或曲西立濱處理之腫瘤 20生長樣本類似（第12A及12B圖）。由於腫瘤的生長不受抑制，於三週後由於腫瘤負擔太大將小鼠安樂死。相反地，使用曲西立濱及崔茲竹美之組合處理可大為顯著抑制腫瘤的生長（第12A圖）。多個腫瘤可作動尺寸的縮小，經過5週處理後7頭小鼠中之4頭不具有可觸診的腫瘤。於使用 112 200835507 RAD001及崔茲竹美處理後，比較單獨RAD〇〇1或崔茲竹美，腫瘤的生長相對緩慢（第12B圖）。如此，崔茲竹美與曲西立濱或RAD001組合可於活體内有效抑制ErbB2過度表現之PTEN缺陷人乳癌異種移植片。 5 已經參照較佳貫施例s兄明本發明。本發明之變化及修改對熟諳技藝人士由前文發明之詳細說明顯然易知。意圖全部此等變化及修改皆係涵蓋於本發明之範圍。【圖式簡單說明3 第1圖顯示由NCI分集集合中識別作為Akt抑制劑候選 10 者之API-2 (曲西立濱）之識別。第1A圖顯示API-2 (曲西立濱）之化學結構式。第1B圖顯示ΑΠ-2抑制於AKT2轉形NIH3T3 細胞中之AKT2磷酸化程度。威爾（Wile)型AKT2-轉形 NIH3T3細胞以API-2 (1 μΜ)處理指示之時間，且接受以抗石粦酸-Akt-T308及-S473抗體（上圖及中圖）之免疫墨點分析。下 15 圖顯示總AKT2之表現。於第1C圖中，顯示API-2抑制Akt 之三個同質異形體。HEK293細胞於EGF刺激前以 HA-Akt 卜 HA-AKT2、及 HA-AKT3 轉移感染且以 API-2 (1 μΜ) 或瓦特曼寧（wortmannin) (15μΜ)處理，細胞經溶解且以 anti-HA抗體免疫沉澱。免疫沉澱物接受試管内激酶檢定分 20 析（上）及以抗磷酸-Akt-T308 (下）抗體進行免疫墨點分析。中圖顯示經轉移感染之Aktl、AKT2、及AKT3之表現。第 1D圖顯示API-2於試管内不會抑制Akt。於試管内組成活性 AKT2重組蛋白質於激酶緩衝液之激酶檢定分析含有ΐμΜ ΑΡΙ_2 (線道3)。 113 200835507 第2圖驗證API-2不會抑制PI3K、PDK1&AGC激誨家族中之緊密相關成員。第2A圖顯示試管内PI3K激酶檢定分析。於EGF刺激前，HEK293細胞經血清匱乏且以API-2 (ΙμΜ)或瓦特曼寧（15μΜ)處理3〇分鐘。細胞經溶解且以 5 anti-p 110 α抗體免疫沉澱。免疫沉澱物使用ΡΙ-4-Ρ作為崦基質接受活體内激酶檢定分析。第2Β圖顯示ΑΡΙ-2對試管内 PDK1活化之效應（上圖），實心圓顯示藉Αρι_2抑制。空心圓顯示藉1¼性對照史妥洛史寶靈（staur〇Sp〇rine)抑制，史妥洛史寶靈為強力PDK1抑制劑(IC50=5 nM)。下圖為HEK293細 10 胞之免疫墨點分析，HEK293細胞於EGF刺激前以 Myc-PDKl轉移感染且以瓦特曼寧或API-2處理。免疫墨點係以指示之抗體檢測。第2C圖顯示PKCa以抗磷酸-PKCa -T638(上圖）及總PKCa (下圖）抗體接著以API-2或非選擇性 PKC抑制劑R〇31_8220處理之磷酸化程度之免疫墨點分 15 析。第2D圖顯示試管内SGK激酶檢定分析。HEK293細胞於 EGF刺激前以HA-SGK轉移感染且以API-2或瓦特曼寧處理。試管試驗之激酶係以HA-SGK免疫沉澱使用MBP作為酶基質進行(上圖）。下圖顯示經轉移感染之HA-SGK之表現。第2E圖顯示PKA激酶檢定分析之結果。經過免疫純化之 20 PKA於含有適用之抑制劑（API-2或PKAI)及酶基質坎普泰 (Kemptide)之ADB緩衝液（上態生技公司（Upstate Biotechnology Inc))中培養。定量激酶活性。於第2F圖中顯示西方墨點。OVCAR3細胞以API-2處理指示時間。細胞溶解產物係以適用之抗磷酸抗體（1-4圖）及抗肌動蛋白抗體 114 200835507 (下圖）進行免疫墨點。第3圖驗證API-2於有升高的Akt之人癌細胞中’抑制 Akt活性及細胞生長而誘導細胞〉周亡。第3 A圖為使用APU 處理後之西方墨點，Akt之鱗酸化程度係以抗磷酸 5 -Akt_T308抗體於適用之人癌細胞系檢測。墨點再度使用抗總Akt抗體探測（下圖）。第3B圖中，顯示細胞增生檢定分析。圖中指示之細胞系以不同劑量之API-2處理24小時及48 小時，然後以細胞力價（CenTiter) 96細胞增生檢定分析套件組（普米嘉公司（Promega))分析。第3C圖提供細胞〉周亡分 10 析。細胞以API-2處理，以附著素（annexin) V及PI染色，及藉FACScan分析。第4圖顯示於小鼠異種移植片中，於有升高之Akt之癌細胞系中，API-2抑制Akt之下游標靶，且具有抗腫瘤活性。於第4A圖中，驗證API-2可抑制結節素(tuberin)、Bad、AFX 15 及GSK-3/3之Akt磷酸化。於以API-2處理後，OVCAR3細胞經溶解且以適用之抗體進行免疫墨點分析。第4B圖顯示 API-2可抑制腫瘤生長。腫瘤細胞注射入裸小鼠體内，左側 Akt細胞濃度低，而右側Akt細胞濃度高。當腫瘤達到約 100-150立方毫米平均大小時，動物以載媒劑或以1毫克/千 20 克/日ApI_2處理。各個測量值表示10個腫瘤之平均值。第4C 圖顯示帶有以API-2或載媒劑（對照組)處理之〇VCAR3 (右）及OVCAR5 (左）之小鼠之代表圖。第4d圖顯示於實驗結束時之腫瘤大小（下）及重量（上)之實例。第4E圖中，腫瘤溶解產物之免疫墨點分析係使用經以API-2處理（T3及T4)及未 115 200835507 經處理（Τ1及T2)之OVCAR-3_衍生腫瘤中之抗碟_Akt-S473 (上）及抗-AKT2 (下）抗體進行。第5圖顯示API-2 (曲西立濱）抑制於試管内抑制Akt激酶活性。試管内激酶檢定分析係使用1>131〇與人]^之重組株 5於含填酸基肌糖醇_3,4,5-p3 (PIP3)、API-2及組織腺H2B作為酶基質之激酶緩衝液進行。培養3〇分鐘後，藉SDS-PAGE 分離反應且暴露於薄膜。第6a-6c圖提供人Aktl之mRNA及胺基酸序列，也標示限剪酶位置。 10 第7a_7d圖提供人Akt2之mRNA及胺基酸序列，也標示限剪酶位置。第8a-8c圖提供人Akt3之mRNA及胺基酸序列，也標示限剪酶位置。第9圖顯示藉崔兹竹美（trastuzuniab)及Akt/mTOR徑路 15抑制劑之組合物之生長抑制。PTEN反訊息或非特異性寡核苷酸轉移感染之BT474.ml細胞單獨以Akt/mTOR徑路抑制劑處理’或以與崔茲竹美之組合物處理，及評估相對細胞生長。第9A圖顯示Akt/mTOR抑制劑之示圖。生長抑制係於經PTEN AS轉移感染之BT474 ml細胞中評估。顯示劑量 20 為：曲西立濱（TCN) ΙμΜ ; RAD001 0.2 nM;QLT〇267 ΙΟμΜ ; KP 372-1 〇·〇5μΜ ; 4ADPIB 5μΜ ;艾朵弗新 (Edelfosine) 7·5μΜ ;及崔茲竹美(Ttzm) 2微克/毫升。指示標準差（SD) ’以生長抑制百分比表示。所示結果為由2_3次實驗所得之組合資料，各實驗中之各項處理重複三次。第 116 200835507 9B圖顯示TCN與崔茲竹美組合抑制細胞生長。BT474.ml細胞以PTEN AS寡核苷酸或非特異性(NS)寡核苷酸轉移感染，以崔茲竹美及TCN於多劑TCN單獨處理或組合處理，且檢定分析生長抑制。崔茲竹美係以單一濃度投予。第9C 5 圖顯示RAD001與崔兹竹美組合抑制細胞生長。BT474.ml 細胞以PTEN AS寡核苷酸或非特異性(NS)寡核苷酸轉移感染，以崔茲竹美及RAD001於多劑RAD001單獨處理或組合處理’且檢定分析生長抑制。用於第9B&9C圖：*指示比較單獨使用崔茲竹美或TCN/TAD001，於組合處理後生長抑 10制之顯者差異。被視為顯著。誤差柱顯示sem。第10圖顯示用於細胞凋亡之協同效果。於接種後24小時，經PTEN AS及NS轉移感染之BT474.ml細胞如所指示以於下列濃度之崔茲竹美(Ttzm)、TCN及/或RAD001處理：崔茲竹美2微克/毫升；曲西立濱2·5μΜ ; RAD001 0.4 nM。進 15行AP0_Brdu土諾(Tunel)檢定分析來評估細胞〉周亡。實驗進行三次’所示資料為平均細胞凋亡。誤差柱顯示標準差。比較全部其它處理，崔茲竹美+曲西立濱處理顯著誘導細胞凋亡（ρ<0·01)。第11圖顯示Akt及P70S6K活性之抑制。為了評估此等藥 20物用MAkt/mT0R徑路之效果，PTEN AS及NS寡核苔酸係於 BT474.ml細胞轉移感染。二日後，細胞以崔茲竹美及曲西立濱（1^^)(第11八圖）或崔兹竹美及11八〇〇〇1(第11_)處理 2小時。收集全細胞溶解產物，藉se>s_PAGE分離及如指示進行免疫墨點分析。崔茲竹美之濃度為2微克/毫升，曲西 117 200835507 立濱為2.5μΜ及RAD001為0.4nM。實驗至少重複兩次來確保結果為可再現性。第12圖顯示於SCID小鼠異種移植片模型中組合治療抑制腫瘤生長。SCID小鼠接受BT474.m 1乳癌細胞異種移植 5片於乳腺脂肪墊。異種移植片成長3週來產生平均大小 100-150立方毫米之腫瘤。投予PTEN反訊息寡核苷酸、崔茲竹美、曲西立濱（第12A圖）及RAD001(第12B圖）。使用測微器每週兩次測量腫瘤大小，對各處理組求取腫瘤大小平均。誤差柱表示平均值之標準差。*指示比較單獨崔茲竹美 10 (Ttzm)、TCN或DMSO，組合治療後之生長抑制有顯著差異。P<0.05被視為顯著。【主要元件符號說明】 (無） 118Almost all of the compounds showed growth inhibition effects in particular at high concentrations and 10 showed growth inhibition effects on cells with MpTEN (Fig. 9). However, when two of the compounds, namely triclinide and RAD〇〇1, were compared with the CuiZhumei group, when Cui class Zhumei or any compound was administered alone, the combination administration could significantly increase the growth inhibition of PTEN AS cells. (Figure 9 eight). Quercetin (also referred to as API-2) is a compound that inhibits Akt activation, and enhances the growth inhibition produced by Triesium 15 in a 20-fold concentration range (Fig. 9B). When RAD001 was administered at a low dose (<1nM), the mTOR inhibitor RAD〇〇1 (Afulololus) increased the growth inhibition produced by Trieste (Fig. 9C). Surprisingly, Quxi Libin and RAD001 can synergize with Tries Zhumei to inhibit cell growth in PTENAS and PTEN NS cells to a similar extent (Fig. 9B and Fig. 9C 20). In other words, Quxi Libin and RAD001 can restore the sensitivity of the PTEN-deficient cells. Tricinem and RAD001 are also effective as single agents in both PTENAS cells and PTEN NS cells, and are greater than 5 nM and 1.5 nM for triclinide and RAD001, respectively (Fig. 9B and Fig. 9C). The third compound, the ILK inhibitor QLT0267, enhanced the growth inhibition produced by Tries Zhumei (Fig. 9A) at a narrow dose range of 109,35,507 (~5-15 pM). The compound was not further studied because of the narrow dose range of synergistic effects between QLT0267 and Cui Sizhu. At concentrations greater than 2〇pM, QLT0267 and Triz Bamboo have no synergistic effect, but a single agent significantly inhibits cell growth. Induction of apoptosis after combination therapy: In order to assess whether growth inhibition is accompanied by apoptosis, the inventors used tromethonine, RAD001, and 崔兹竹美 alone or in combination to treat BT474 transfected with PTEN AS and NS. .ml cells, and quantify the degree of apoptosis (Figure 1). RAD001 alone 10 alone or in combination with Tries Zhumei did not significantly induce apoptosis. Although the number of TUNEL-positive apoptotic cells increased slightly after treatment with Trizut or Trifluralin alone, this increase was not statistically significant. However, compared with all other treatments in both PTEN AS and NS-transfected cells, the combination of tricineribine and triazepam significantly induced apoptosis (Fig. 10). 15 Example 6: TCN inhibits activation of Akt and mTOR inhibition by downstream signaling molecules: Immunoblotting analysis confirmed that triclinide and RAD001 block Akt and mTOR (an important signaling molecule by ErbB2 activation) and Quxili Activation of the targets of Bin and RAD001. Akt was used for phosphorylation of Thr308 and Ser473 as an indicator of Akt activity; mTOR activity was assessed by phosphorylation of p70S6K20 (70-kDa ribosomal protein S6 kinase), also known as mTOR target. After treatment with trifluralin, Akt phosphorylation was substantially reduced at two locations (Fig. 11A). In PTEN-deficient cells, the degree of phosphorylation of Akt was similar to that seen in cells with intact pteN after treatment with tresibine and Cui Zhumei (Fig. 11A's lanes 4 and 8). Thus, Querceptine overcomes the adverse effects of PTEN loss by effectively blocking Akt activity. RAD001 significantly blocked the phosphorylation of P70S6K (Fig. 11B). However, the combination of RAD001 and Tries Zhumei does not reduce P70S6K phosphorylation as seen by RAD001 alone (Fig. 11B). Late recognition of the feedback loop, which can lead to phosphorylation and activation of Akt after treatment with mTOR inhibitor 5 such as RAD001 (O'Reilly KE et al., Cancer Research, 2006. 66(3): P.1500-8) . It has also been observed that the use of RAD001 and the combination of Triz Zhumei and RAD001 to treat activated Akt can eliminate Akt phosphorylation by this feedback loop (Fig. 11B, lane 3 vs. lane 4), and mTOR The inhibition of Akt activation is consistent with the upstream receptor tyrosine kinase 10 (〇, Reilly KE et al., Cancer Research, 2006. 66(3): P.1500-8). In other words, the drug inhibits its predicted target kinase, and the combination therapy has a greater inhibitory effect on the Akt/mTOR signaling pathway than any single agent', even in PTEN-deficient cells. Example 7: Inhibition of tumor growth in PTEN-deficient tumor TCN and Cui Zhemei 15 Materials and methods Human tumor research model in SCID mouse xenograft: female, 6_week old, severe combined immunodeficiency (SCID) mouse line Tac〇nic Farm (Hudson, NY). Tumor xenografts are performed as described by Nagata Y et al., Cancer Cell, 2004, 6(2): ρ·117-27. When xenograft tumors reached an average size of 2〇100-150 mm3, the mice were divided into 6 groups, each group had 7 mice and the average distribution of tumor size, and was treated as described later. PTEN anti-message (30 pg) nucleus citrate was administered to each mouse every week by intratumoral injection. The treatment of the drug was started one week after the PTEN AS nucleation acid administration.崔兹竹美 was administered at a dose of G·5 mg/kg twice a week in an intratumoral injection of multiple parts 111, 200835507. The CuiZhumei system was administered intraperitoneally (ι·ρ·) at a dose of 0.5 mg/kg/day in 200 pL of 20% DMSO saline solution. RAD001 is administered twice a week through a gastric tube at a dose of 1 picogram per kilogram in 500 picoliters of 5% glucose water. 20% DMSO saline solution (200 picolitres 5 / day) was administered by intraperitoneal injection. Tumor size was measured twice a week using a micrometer, and the tumor volume was calculated as volume = length X width 2/2. Statistical analysis: One-way ANOVA was performed using GraphPad Prism 3·〇 for Windows (Graph Pad Software, San Diego, CA). 10 Results Early biometric and molecular data are extremely promising, but in vivo studies provide the most demanding tests for therapeutic effects. Therefore, Quxi Libin and RAD〇〇1 were tested in vivo. BT474.ml cell xenografts were injected into the breast fat pad of 6 week old SCID mice. After tumor formation, mice received PTEN AS via a tumor within 15 tumors per week. This regimen is effective as a model of PTEN-deficient tumors in vivo (Nagata γ· et al., Cancer cells, 2_, 6(2): Ρ·117-27). Mice were randomized into multiple treatment groups receiving either triclinib, RAD00, or DM00 alone or in combination. After treatment, tumor 20 growth samples treated with DMSO, Cui Zhemei, Shakuhachi: 〇001 or triclinide alone were similar (Figures 12A and 12B). Since the growth of the tumor was not inhibited, the mice were euthanized after three weeks due to the large tumor burden. Conversely, treatment with a combination of triclinide and 崔兹竹美 can significantly inhibit tumor growth (Fig. 12A). Multiple tumors were reduced in size, and 4 of the 7 mice did not have palpable tumors after 5 weeks of treatment. After treatment with 112 200835507 RAD001 and Cui Zhemei, the tumor growth was relatively slow compared to RAD〇〇1 or Cui Zhemei alone (Fig. 12B). Thus, the combination of Triz Chumei and Qucilide or RAD001 can effectively inhibit the excessive expression of ErbB2 in vivo in PTEN-deficient human breast cancer xenografts. 5 The invention has been described with reference to preferred embodiments. Variations and modifications of the present invention are apparent to those skilled in the art from the foregoing detailed description of the invention. All such changes and modifications are intended to be included within the scope of the present invention. [Simple Description of the Drawings 3 Figure 1 shows the identification of API-2 (Quxi Libin) identified as a candidate for Akt inhibitors in the NCI diversity set. Figure 1A shows the chemical structure of API-2 (Quxi Libin). Figure 1B shows that ΑΠ-2 inhibits the degree of AKT2 phosphorylation in AKT2 transduced NIH3T3 cells. Will-type AKT2-transformed NIH3T3 cells were treated with API-2 (1 μΜ) for the indicated time and received immunostaining with anti-phosphinic acid-Akt-T308 and -S473 antibodies (top and bottom) Point analysis. Figure 15 below shows the performance of total AKT2. In Figure 1C, three isomorphs of API-2 inhibition of Akt are shown. HEK293 cells were infected with HA-Akt, HA-AKT2, and HA-AKT3 prior to EGF stimulation and treated with API-2 (1 μΜ) or wortmannin (15 μΜ). Cells were lysed and anti-HA Antibody immunoprecipitation. Immunoprecipitates were subjected to in vitro assays for kinase assays (upper) and immunoglobulin assays with anti-phospho-Akt-T308 (bottom) antibodies. The middle panel shows the performance of Aktl, AKT2, and AKT3 by transfer infection. Figure 1D shows that API-2 does not inhibit Akt in the test tube. The kinase assay for the active AKT2 recombinant protein in a kinase buffer containing ΐμΜ ΑΡΙ_2 (lane 3) was constructed in vitro. 113 200835507 Figure 2 verifies that API-2 does not inhibit closely related members of the PI3K, PDK1 & AGC family. Figure 2A shows the PI3K kinase assay in vitro. HEK293 cells were serum-deficient and treated with API-2 (ΙμΜ) or Watmanning (15 μM) for 3 min before EGF stimulation. The cells were lysed and immunoprecipitated with 5 anti-p 110 α antibody. Immunoprecipitates were assayed for in vivo kinase assays using ΡΙ-4-Ρ as the thiol matrix. Figure 2 shows the effect of ΑΡΙ-2 on the activation of PDK1 in the test tube (above), and the solid circle shows inhibition by Αρι_2. The open circle showed inhibition by the 11⁄4 sex control, Staur〇Sp〇rine, which was a potent PDK1 inhibitor (IC50 = 5 nM). The following figure shows the immunoblotting analysis of HEK293 cells. HEK293 cells were infected with Myc-PDK1 prior to EGF stimulation and treated with Watmanin or API-2. The immune dot is detected by the indicated antibody. Figure 2C shows the immunostaining of PKCa with anti-phospho-PKCa-T638 (top panel) and total PKCa (lower panel) antibodies followed by API-2 or non-selective PKC inhibitor R〇31_8220. Analysis. Figure 2D shows an in vitro SGK kinase assay. HEK293 cells were infected with HA-SGK transfer prior to EGF stimulation and treated with API-2 or Watmanin. The test tube kinase was immunoprecipitated with HA-SGK using MBP as the enzyme substrate (top panel). The figure below shows the performance of HA-SGK transfected with infection. Figure 2E shows the results of the PKA kinase assay. The immunopurified 20 PKA was cultured in ADB buffer (Upstate Biotechnology Inc) containing the appropriate inhibitor (API-2 or PKAI) and the enzyme substrate Kemptide. Quantify kinase activity. Western blots are shown in Figure 2F. OVCAR3 cells were timed with API-2 treatment. The cell lysate was immunized with the appropriate anti-phospho antibody (1-4) and anti-actin antibody 114 200835507 (bottom panel). Figure 3 demonstrates that API-2 induces cell death by inhibiting Akt activity and cell growth in human cancer cells with elevated Akt. Figure 3A shows the western blot after treatment with APU. The degree of squamization of Akt is detected by anti-phospho-5-Akt_T308 antibody in a suitable human cancer cell line. The ink spots were again probed with anti-total Akt antibody (bottom panel). In Fig. 3B, a cell proliferation assay is shown. The cell lines indicated in the figure were treated with different doses of API-2 for 24 hours and 48 hours, and then analyzed by the CenTiter 96 Cell Proliferation Assay Kit (Promega). Figure 3C provides the analysis of cells > weeks of death. Cells were treated with API-2, stained with annexin V and PI, and analyzed by FACScan. Figure 4 shows that in mouse xenografts, API-2 inhibits Akt downstream targets in cancer cell lines with elevated Akt and has antitumor activity. In Figure 4A, it was verified that API-2 inhibited Akt phosphorylation of tuberin, Bad, AFX 15 and GSK-3/3. After treatment with API-2, OVCAR3 cells were lysed and subjected to immunoblot analysis with applicable antibodies. Figure 4B shows that API-2 inhibits tumor growth. Tumor cells were injected into nude mice with a low concentration of Akt cells on the left and a high concentration of Akt cells on the right. When the tumor reached an average size of about 100-150 cubic millimeters, the animals were treated with vehicle or at 1 mg/kg 20 g/day of ApI_2. Each measurement represents the average of 10 tumors. Figure 4C shows a representation of mice with sputum VCAR3 (right) and OVCAR5 (left) treated with API-2 or vehicle (control). Figure 4d shows an example of tumor size (bottom) and weight (top) at the end of the experiment. In Figure 4E, the immunoblot analysis of tumor lysates was performed using anti-disc_Akt in OVCAR-3_derived tumors treated with API-2 (T3 and T4) and not 115 200835507 (Τ1 and T2). -S473 (top) and anti-AKT2 (bottom) antibodies were performed. Figure 5 shows that API-2 (Cucidine) inhibits Akt kinase activity in vitro. The in-tube kinase assay uses 1>131〇 and human recombinant strain 5 in the acid-containing inositol_3,4,5-p3 (PIP3), API-2 and tissue gland H2B as the enzyme substrate. Performed in kinase buffer. After 3 min incubation, the reaction was separated by SDS-PAGE and exposed to the membrane. Figures 6a-6c provide mRNA and amino acid sequences for human Aktl and also indicate the position of the restriction enzyme. 10 Figure 7a_7d provides the mRNA and amino acid sequence of human Akt2, also indicating the position of the restriction enzyme. Figures 8a-8c provide mRNA and amino acid sequences of human Akt3, also indicating the position of the restriction enzyme. Figure 9 shows the growth inhibition by a combination of trastuzuniab and Akt/mTOR pathway inhibitors. BT474.ml cells infected with PTEN anti-message or non-specific oligonucleotides were treated with Akt/mTOR pathway inhibitor alone or treated with a composition of Triz, and relative cell growth was assessed. Figure 9A shows a diagram of an Akt/mTOR inhibitor. Growth inhibition was assessed in BT474 ml cells infected with PTEN AS transfer. The dose 20 is shown as: triclinide (TCN) ΙμΜ; RAD001 0.2 nM; QLT〇267 ΙΟμΜ; KP 372-1 〇·〇5μΜ; 4ADPIB 5μΜ; Edelfosine 7·5μΜ; Beauty (Ttzm) 2 μg/ml. The indicated standard deviation (SD)' is expressed as a percentage of growth inhibition. The results shown are the combined data obtained from 2 to 3 experiments, and each treatment in each experiment was repeated three times. No. 116 200835507 9B shows that TCN combined with Tries Zhumei inhibits cell growth. BT474.ml cells were infected with PTEN AS oligonucleotides or non-specific (NS) oligonucleotides, treated with Trizin and TCN in multiple doses of TCN alone or in combination, and assayed for growth inhibition. Triz Bamboo is administered at a single concentration. Figure 9C 5 shows that RAD001 combined with Tries Zhumei inhibits cell growth. BT474.ml cells were infected with PTEN AS oligonucleotides or non-specific (NS) oligonucleotides, treated with Trizut and RAD001 alone or in combination with multiple doses of RAD001' and assayed for growth inhibition. For the 9B & 9C chart: * indicates a significant difference in the growth inhibition system after combination treatment using Trizut or TCN/TAD001 alone. It is considered significant. The error bar shows sem. Figure 10 shows the synergistic effect for apoptosis. At 24 hours after inoculation, BT474.ml cells infected with PTEN AS and NS were treated as indicated by Ttzm, TCN and/or RAD001 at the following concentrations: Trizum 2 μg/ml; Quxi Libin 2·5μΜ; RAD001 0.4 nM. Into 15 rows of AP0_Brdu Tunel assay to assess cell death. The data shown in the experiment three times was average apoptosis. The error bars show the standard deviation. Comparing all other treatments, Cui Zhemei + Quciliden significantly induced apoptosis (ρ < 0·01). Figure 11 shows inhibition of Akt and P70S6K activity. In order to evaluate the effect of these drugs on the MAkt/mT0R pathway, PTEN AS and NS oligosaccharide were in BT474.ml cell transfer infection. Two days later, the cells were treated with Cuiz Bamboo and Quxi Libin (1^^) (Fig. 11) or Cui Zhumei and 11 Oct. 1 (11_) for 2 hours. Whole cell lysates were collected, separated by se>s_PAGE and subjected to immunoblot analysis as indicated. The concentration of Cuiz Bamboo is 2 μg/ml, Quxi 117 200835507 Libin is 2.5 μΜ and RAD001 is 0.4 nM. The experiment is repeated at least twice to ensure that the result is reproducible. Figure 12 shows that combination therapy inhibits tumor growth in a SCID mouse xenograft model. SCID mice received BT474.m 1 breast cancer xenografts 5 tablets in the mammary fat pad. Xenografts were grown for 3 weeks to produce tumors with an average size of 100-150 mm3. PTEN anti-information oligonucleotides, Cui Zhemei, Quxi Libin (Fig. 12A) and RAD001 (Fig. 12B) were administered. Tumor size was measured twice a week using a micrometer, and tumor size was averaged for each treatment group. The error bars represent the standard deviation of the mean. *Indications comparing Tzm, TCN or DMSO alone, there was a significant difference in growth inhibition after combination therapy. P < 0.05 was considered significant. [Main component symbol description] (none) 118

Claims

200835507 十、申請專利範圍： 1. 一種組成物，包含： ⑴式I-IV化合物：200835507 X. Patent application scope: 1. A composition comprising: (1) a compound of formula I-IV:

oh3Oh3

:Ηι:Ηι

其中R2’、R3’及R5’分別為氫、視需要可經取代之磷酸根或膦酸根（包括一磷酸、二磷酸、或三磷酸或安定 10 化磷酸前藥）；醯基（包括低碳醯基）；烷基（包括低碳烷基）；si胺、績酸s旨包括烧基或芳基烧基；績醯基包括甲磺醯基及苄基，其中該苯基視需要可經以一個或多個 119 200835507 如於此處所述芳基之定義中說明之取代基取代；視需要可經取代之芳基磺醯基；脂質包括磷脂質；胺基酸；碳水化合物；胜肽；或膽固醇；或其它藥學上可接受之離去基，其於活體内可提供一種化合物其中r2,、r3’或R5’ 分別為Η或一填酸、二磷酸、或三填酸；其中Rx及Ry分別為氫、視需要可經取代之磷酸根；醯基（包括低碳醯基）；醯胺、烷基（包括低碳烷基）；芳香族、去氧伸烧基諸如聚乙二醇、視需要可經取代之芳基磺醯基；脂質包括磷脂質；胺基酸；碳水化合物；胜肽；或膽固醇；或其它藥學上可接受之離去基；於一個實施例中，該化合物係呈5，-磷醚脂質或5，_醚脂質投藥； Ri及R2各自分別為Η、視需要可經取代之直鏈、分支、或環狀烧基（包括低碳烧基）、稀基或炔基、C〇-烧基、C0_稀基、c〇_炔基、c〇_芳基或雜芳基、c〇_炫氧基烷基、CO-芳氧基烷基、CO-經取代之芳基、磺醯基、烷基磺醯基、芳基磺醯基、芳烷基磺醯基； (H)表皮生長因子受體抑制劑或其鹽；以及 (iii)藥學上可接受之載劑。士申明專利範圍第丨項之組成物，其中式化合物為曲西立濱（tdcidbine)、磷酸曲西立濱、磷酸曲西立濱一水合物或其組合。士申π專利範圍第丨項之組成物，其中該表皮生長因子文體抑制劑為傑費堤尼(gefitinib)。如申π專利範圍第i項之組成物，其中該表皮生長因子 120 200835507 受體抑制劑為爾洛堤尼(erlotinib)。 5.如申請專利範圍第！項之組成物，其中該式ι化合物係以至少20毫克/平方米之劑量存在。 6·如申請專利範圍第i項之組成物，其中該式1化合物係以至少10毫克/平方米之劑量存在。 7·如申請專利範圍第i項之組成物，其係適合供經投予。 8. 如申請專利_第7項之組成物，其巾該腸道經靜脈投予。々 10 15 20 9. 如申請專利範圍第i項之組成物，其係適合供經口 ^如申請專職圍第1項之組成物，其係適合供局部 11.如申請專利範圍第丨項之組成予。 . 其中該崔茲竹1 (trastuziimab)或其鹽係以由約i臺古，τ ^ 量存在。約嶋毫克之數 12·如申請專利範圍第1項之組成物，其中飞崔# 13 鹽係以由約1〇〇毫克至約500毫克之數旦 4竹美或其 •如申請專利範圍第1項之組成物，其里存在。鹽係以由約200毫克至約450毫克之數旦4*從竹美或其 14·如申請專利範圍第1項之組成物，其里^在^ 鹽係以約440毫克之數量存在。輕⑼竹美或其 15· 一種於一哺乳動物治療腫瘤或癌症乳動物投予有效量之一種組成物包含：匕3對该哺 ⑴式I-IV化合物·· 121 200835507Wherein R2', R3' and R5' are respectively hydrogen, optionally substituted phosphate or phosphonate (including monophosphate, diphosphate, or triphosphate or diazepam phosphate prodrug); sulfhydryl (including low carbon) Alkyl (including lower alkyl); si amine, acid s is intended to include alkyl or arylalkyl; methane includes methanesulfonyl and benzyl, wherein the phenyl can be used as needed Substituted by one or more of 119 200835507 as described in the definition of aryl as described herein; optionally substituted arylsulfonyl; lipids including phospholipids; amino acids; carbohydrates; peptides Or cholesterol; or other pharmaceutically acceptable leaving group, which provides a compound in vivo wherein r2, r3' or R5' are respectively hydrazine or an acid-filled, di-phosphoric acid, or tri-acid; And Ry are respectively hydrogen, optionally substituted phosphate; sulfhydryl (including low carbon fluorenyl); decylamine, alkyl (including lower alkyl); aromatic, deoxyalkylene such as polyethylene Alcohol, optionally substituted arylsulfonyl; lipids including phospholipids; amine groups Carbohydrate; peptide; or cholesterol; or other pharmaceutically acceptable leaving group; in one embodiment, the compound is administered as a 5,-phospho-lipid or 5,-ether lipid; Ri and R2 are each a linear, branched, or cyclic alkyl group (including a low carbon alkyl group), a dilute or an alkynyl group, a C〇-alkyl group, a C0_thin group, a c〇-alkynyl group, which may be substituted as needed. C〇_aryl or heteroaryl, c〇_decyloxyalkyl, CO-aryloxyalkyl, CO-substituted aryl, sulfonyl, alkylsulfonyl, arylsulfonyl An aralkylsulfonyl group; (H) an epidermal growth factor receptor inhibitor or a salt thereof; and (iii) a pharmaceutically acceptable carrier. The composition of the ninth aspect of the invention, wherein the compound is tdcidbine, triclinbium phosphate, triclinbine phosphate monohydrate or a combination thereof. The composition of the third aspect of the patent application, wherein the epidermal growth factor streptozotoxin is gefitinib. The composition of the item i of claim π, wherein the epidermal growth factor 120 200835507 receptor inhibitor is erlotinib. 5. If you apply for a patent range! A composition of the formula wherein the compound of formula ι is present in a dosage of at least 20 mg/m 2 . 6. The composition of claim i, wherein the compound of formula 1 is present in a dose of at least 10 mg/m 2 . 7. If the composition of the item i of the patent application is applied, it is suitable for administration. 8. If the composition of the patent_Article 7 is applied, the intestine is administered intravenously. 々10 15 20 9. If the composition of the scope of the application of the scope of the patent item i is suitable for oral administration, for example, the composition of the first item of the full-time enclosure is suitable for the part 11. For example, the scope of the patent application Composition. Among them, the trastuziimab or its salt system exists in the quantity of about i. The number of milligrams is 12. The composition of the first paragraph of the patent application, wherein the Fei Cui # 13 salt is from about 1 gram to about 500 milligrams of the number of 4 bamboo or its • as claimed The composition of item 1 exists in it. The salt is a composition of the first item of the patent range from about 200 mg to about 450 mg of a denier 4*, wherein the salt is present in an amount of about 440 mg. Light (9) Zhumei or 15 thereof. A composition for treating a tumor or cancer in a mammal. An effective amount of a composition comprising: 匕3 to the feeding (1) Formula I-IV Compound·· 121 200835507

9¾93⁄4

MM

其中R2’、R3’及R5’分別為氫、視需要可經取代之磷酸根或膦酸根（包括一磷酸、二磷酸、或三磷酸或安定化磷酸前藥）；醯基（包括低碳醯基）；烷基（包括低碳烷基）；醯胺、磺酸酯包括烷基或芳基烷基；磺醯基包括甲磺醯基及苄基，其中該苯基視需要可經以一個或多個如於此處所述芳基之定義中說明之取代基取代；視需要可經取代之芳基磺醯基；脂質包括磷脂質；胺基酸；碳水化合物；胜肽；或膽固醇；或其它藥學上可接受之離 122 10 200835507 去基’其於活體内可提供一種化合物其中r/、R;，或I，分別為Η或一磷酸、二磷酸、或三磷酸；其中R及Ry分別為氫、視需要可經取代之鱗酸根；醯基（包括低碳醯基）；醯胺、烷基（包括低碳烷基）；芳 5 香族、去氧伸烷基諸如聚乙二醇、視需要可經取代之芳基磺酸基；脂質包括磷脂質；胺基酸；碳水化合物；胜肽；或膽固醇；或其它藥學上可接受之離去基；於一個貫施例中’該化合物係呈5，_鱗醚脂質或5，_醚脂質投藥； Rl及R2各自分別為Η、視需要可經取代之直鏈、分 10 支、或環狀烷基（包括低碳烷基）、烯基或炔基、CO-烷基、CO-稀基、co_炔基、c〇_芳基或雜芳基、c〇_烧氧基烧基、CO-芳氧基烧基、CO-經取代之芳基、橫醯基、烷基磺醯基、芳基磺醯基、芳烷基磺醯基； (H)表皮生長因子受體抑制劑或其鹽。 15 I6·如申請專利範圍第丨5項之方法，其中該式Ι-ΐν化合物及 "亥表皮生長因子受體抑制劑或其鹽係同時投予。 Π·如申請專利範圍第15項之方法，其中該式I-IV化合物係於崔兹竹美或其鹽投予之後投予。 ^ 18·如巾請專利範圍第15項之方法，其中該表皮生長因子受 2〇體抑咖或其龍权後接著奸式I.IV化合物。 19·如申請專利範圍第15項之方法，其中該式I-IV化合物係 :k技予-人计二週，接著為未投予任何化合物之一週時間。 2〇·如申請專利範圍第15項之方法，其中該表皮生長因子受 123 200835507 體抑制劑或其鹽係每週投予一次計三週，接著為未投予任何化合物之一週時間。 21.如申請專利範圍第20項之方法，其中該投藥計劃係重複至少兩次。 5 22.如申請專利範圍第20項之方法，其中該投藥計劃係重複至少四次。 23. 如申請專利範圍第15項之方法，其中該接受治療之腫瘤為***腫瘤、胰臟腫瘤、卵巢腫瘤及大腸直腸腫瘤。 24. 如申請專利範圍第15項之方法，其中投予至少10毫克/ 10 平方米式I_IV化合物。 25. 如申請專利範圍第15項之方法，其中投予至少100毫克表皮生長因子受體抑制劑或其鹽。 26. 如申請專利範圍第15項之方法，其中投予至少200毫克表皮生長因子受體抑制劑或其鹽。 15 27.如申請專利範圍第15項之方法，其中投予至少400毫克表皮生長因子受體抑制劑或其鹽。 124Wherein R2', R3' and R5' are respectively hydrogen, optionally substituted phosphate or phosphonate (including monophosphate, diphosphate, or triphosphate or stabilized phosphate prodrug); sulfhydryl (including low carbon hydrazine) Alkyl (including lower alkyl); decylamine, sulfonate includes alkyl or arylalkyl; sulfonyl includes methanesulfonyl and benzyl, wherein the phenyl can be used as needed Or a plurality of substituents as described in the definition of aryl groups described herein; optionally substituted arylsulfonyl groups; lipids including phospholipids; amino acids; carbohydrates; peptides; or cholesterol; Or other pharmaceutically acceptable excipients from 122 10 200835507 to provide a compound wherein r/, R;, or I, respectively, is hydrazine or monophosphate, diphosphate, or triphosphate; wherein R and Ry Respectively hydrogen, optionally substituted sulphate; sulfhydryl (including lower sulfhydryl); decylamine, alkyl (including lower alkyl); aromatic 5, deoxyalkylene such as polyethylene Alcohol, optionally substituted aryl sulfonate; lipids including phospholipids; amine groups Carbohydrate; peptide; or cholesterol; or other pharmaceutically acceptable leaving group; in one embodiment, the compound is administered as a 5,-squaternary ether lipid or a 5,-ether lipid; Rl and R2 are each A linear, 10 or a cyclic alkyl group (including a lower alkyl group), an alkenyl group or an alkynyl group, a CO-alkyl group, a CO-dense group, a co-alkynyl group, respectively, which may be substituted. , c〇_aryl or heteroaryl, c〇_alkyloxyalkyl, CO-aryloxyalkyl, CO-substituted aryl, hydrazino, alkylsulfonyl, arylsulfonyl a arylalkylsulfonyl group; (H) an epidermal growth factor receptor inhibitor or a salt thereof. 15 I6. The method of claim 5, wherein the Ι-ΐν compound and the "Hyptian epidermal growth factor receptor inhibitor or a salt thereof are administered simultaneously. The method of claim 15, wherein the compound of the formula I-IV is administered after the administration of Triz or its salt. The method of claim 15, wherein the epidermal growth factor is subjected to a compound of the formula I.IV. 19. The method of claim 15, wherein the compound of formula I-IV is: k technology for two weeks, followed by one week without any compound being administered. 2. The method of claim 15, wherein the epidermal growth factor is administered once a week for three weeks by the 123 200835507 body inhibitor or a salt thereof, followed by one week of not administering any compound. 21. The method of claim 20, wherein the dosing schedule is repeated at least twice. 5 22. The method of claim 20, wherein the dosing plan is repeated at least four times. 23. The method of claim 15, wherein the treated tumor is a breast tumor, a pancreatic tumor, an ovarian tumor, and a colorectal tumor. 24. The method of claim 15, wherein at least 10 mg / 10 square meters of the compound I_IV is administered. 25. The method of claim 15, wherein at least 100 mg of an epidermal growth factor receptor inhibitor or a salt thereof is administered. 26. The method of claim 15, wherein at least 200 mg of an epidermal growth factor receptor inhibitor or a salt thereof is administered. The method of claim 15, wherein at least 400 mg of an epidermal growth factor receptor inhibitor or a salt thereof is administered. 124