TW200831525A

TW200831525A - Compositions including triciribine and one or more platinum compounds and methods of use thereof

Info

Publication number: TW200831525A
Application number: TW096146510A
Authority: TW
Inventors: Lawrence Akinsanmi
Original assignee: Vioquest Pharmaceuticals Inc
Priority date: 2006-12-06
Filing date: 2007-12-06
Publication date: 2008-08-01
Also published as: WO2008070136A1

Abstract

This application encompasses combination therapies including triciribine and related compounds and one or more platinum compounds and compositions with reduced toxicity for the treatment and prevention of tumors, cancer, and other disorders associated with abnormal cell proliferation.

Description

200831525 九、發明說明：【發明所屬之技術領域】本案請求美國臨時專利申請案第60/872,990號，申請曰 2006年12月6曰之權益，該案全文以引用方式併入此處。 • 5 發明領域本發明係關於具有較低毒性用於腫瘤、癌症及其它與 … 異常細胞增生相關病症之治療及預防之包括曲西立濱化合物及一種或多種鉑化合物之組合治療及組成物。 # 【先前技術】 10 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，無法適當發揮功犯，且可能傳播至身體的多個區域。組織的異常 ; 15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、入侵且摧毀鄰近的異常組織且傳播遍及身體。癌症可根據其來源體液種類或組織種類而歸類，或可 20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。癌瘤為身體組織中稱作為上皮組織，上皮組織覆蓋或襯塾於器官、腺體或身體結構表面之癌症。舉例言之，胃内概的 5 200831525 癌症稱作為癌瘤。多種癌瘤影響涉及分泌之裔s或腺體，諸如製造乳汁的乳腺。癌射食部癌症病例之80%至90%。肉瘤為由結締組織諸如軟骨、脂肪、肌肉、肌鍵、及硬骨所長出的惡性腫瘤。最常見的肉濟是長在硬骨上的腫瘤’ 5常見於年輕成人。肉瘤之實例包拉骨肉瘤(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 10 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。白血病之實例包括急性骨髓性白血病、慢性骨髓性白血病、急性淋巴細胞性白血病、及慢性淋巴細胞性白血病。骨髓性 15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—根硬骨而形成單一腫瘤稱作為漿細胞瘤。但於其它病例中，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。 20 賴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^瘤阻遏子基因的去活化，以及包括細胞生長促進基因或致癖基因的活化。今日於動物模型中已經識別出數百種活化: 細胞致癌基因，但已經證實只有極少數基因係與人類癌症 200831525 - 5 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，199〇, 63 .p 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· pp 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。 10 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，20〇〇 100(1): ρ· 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 " 15 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/Raf7Mek/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/3及其受體之表現降低），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 20 路的過度活化），d)持續血管新生（亦即VEGF之分泌濃度高）以及〇組織入侵及轉移（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· 57_700)。受體酪胺酸激酶諸如EGFR、ErbB2、VEGFR及胰島素 7 200831525</ RTI> </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> </ RTI> </ RTI> </ RTI> </ RTI> <RTIgt; • 5 FIELD OF THE INVENTION The present invention relates to combination therapies and compositions comprising tricineridine compounds and one or more platinum compounds having lower toxicity for the treatment and prevention of tumors, cancer and other disorders associated with abnormal cell proliferation. # [Prior Art] 10 Background of the Invention Cancer is an abnormal growth of cells. Cancer cells replicate rapidly despite spatial 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 tissue; 15 growth is referred to as a cluster of cells that are 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 adjacent abnormal tissues and spread throughout the body. Cancer can be classified according to the type of body fluid or tissue it is derived from, or can be classified according to 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. Carcinoma is a cancer in the body tissue called epithelial tissue that covers or lining the surface of organs, glands, or body structures. For example, in the stomach, 5 200831525 cancer is called cancer. A variety of cancerous effects involve the secretion of s or glands, such as the mammary glands that make milk. 80% to 90% of cancer cases in the cancer injection department. Sarcomas are malignant tumors that are grown by connective tissues such as cartilage, fat, muscle, muscle bonds, and hard bones. The most common form of meat is a tumor that grows on a hard bone. 5 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, which is responsible for the production of white blood cells and cleansing body fluids or lymphomas 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 is also known as 10. 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 diseases include acute myeloid leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia, and chronic lymphocytic leukemia. The terms bone marrow 15 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 hard bone to form a single tumor called a plasmacytoma. In other cases, however, myeloma cells are aggregated into multiple hard bones to form multiple bone tumors. Called as multiple myeloma. 20 Lai induction and progression are often the result of changes in the accumulation of tumor cell genomes. Such changes may include deactivation of a cell growth inhibitory factor or a tumor repressor gene, as well as activation of a cell growth promoting gene or a sputum gene. Hundreds of activations have been identified in animal models today: cell oncogenes, but only a very small number of genes have been shown to be associated with human cancer 200831525-5 (Weinberg et al., the molecular cause of oncogenes and cancer, 1989, Cold Spring Harbor Experiment Room, New York; Stanbridge J. et al., Cell, 199〇, 63.p 867-874; Godwin et al., Gynecologic Oncology: Principles and Practice, Hoskins WJ” Perez CA· and Young RC· (eds.), 1992· Pp 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 cause a number of cellular effects, such as this Factors may lead to overexpression of gene products. Several oncogenes involved in human cancers can be activated by over-expression of cytokines. 10 It is clear that the continuous gene disorder obtained by cancer genes leads to the regulation of normal cell proliferation, differentiation and planning. Defects in the regulation of apoptosis in signal transduction pathways (Hanahan, D. and Weinberg RA·, cells, 20〇〇100(1): ρ· 57-700). This leads to the basic defects of cell physiology leading to malignant diseases. These defects include: a) self-deficiency of growth signals (ie, growth factors & 15 sub-receptor tyrosine kinases such as EGFR overexpression, And downstream signal transduction pathways such as Ras/Raf7Mek/Erk 1/2 and Ras/PI3K/Akt are out of sequence), b) resistant to growth signals (ie, decreased expression of TGF/3 and its receptors) c) evade apoptosis (ie, loss of pre-apoptotic p53; excessive expression of pre-existing Bcl-2; survival pathway such as PI3K/Akt-mediated survival of 20 pathways of overactivation), d) persistence Angiogenesis (ie, high secretion of VEGF) and invasion and metastasis of sputum tissue (ie, extracellular protease and pre-transfer lignin) (Hanahan, D. and Weinberg RA, Cell, 2000·100(1): ρ· 57_700) Receptor tyrosine kinases such as EGFR, ErbB2, VEGFR and insulin 7 200831525

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 K等人，癌症發生，2004· 25(2); ρ·241-8; SekharamM·等人，癌症研究，2003, 63(22): p.7708-16)。配體諸如EGF、VEGF 5及IGF_1結合至其受體，促成特有酪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 (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）·· 10 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 15 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)。The series of growth factor I receptors (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 K et al., Cancer Occurrence, 2004. 25(2); ρ· 241-8; SekharamM et al., Cancer Research, 2003, 63(22): p. 7708-16). The binding of ligands such as EGF, VEGF 5 and IGF_1 to their receptors contributes to the stimulation of specific tyrosine kinase activity, the autophosphorylation of specific tyrosines at the cytoplasmic functional site of the receptor and the triggering of many different complex signal transductions. Recruitment of signaling proteins for pathways (Olayioye MA et al., Embo J, 2000· 19(13): ρ·3159-67; Porter AC· and Vaillancourt RR·, Oncogenes, 1998·17 (11 )··· 10 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 ovarian cancer, the path of Akt media has been shown to be critical in multiple steps of malignant transformation, malignant transformation. The steps include fine 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 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-20 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，PIP3將Akt募集至漿膜，於漿膜，Akt藉於 Thr308及Ser473 (Aktl)、Thr308及Ser474 (Akt2)及Thr308及 Ser472 (Akt3) (Datta S.R·等人，基因Dev，1999· 13(22): p.2905-27)之填酸化活化。如此，PI3K係經由構酸化PIP2 200831525 " 5 r • 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。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·29〇5-27 ; BellacosaA·等人，國際癌症期刊，1995. 64(4): p.280-5 ; Sim Μ.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/或高度活化(Roy H.K.等人，癌症發生，2002. 23(1): ρ·201_5 ; Altomare D.A.等人，J細胞Biochem，2003. 88(1): ρ· 10 470-6 ; Sun Μ·等人，癌症研究，2001· 61(16): P.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): ρ· 9267-71; 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 15 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); ρ·241-8 ; Sekharam M·等人，癌症研究， 2003，63(22): ρ·7708·16 ; Cohen B.D.等人，Biochem Soc Symp，1998.63: p.199-210 ; Muller W.J·等人，Biochem Soc 20 Symp，1998.63: p.149-57 ; Miller W.E·等人，J Virol，1995· 69(7): ρ·4390-8 ; SlamoiiDj·等人，科學，1987. 235(4785): p.177-82 ; Andmlis.LL·等人，j Clin Oncol，1998.16(4): p.1340-9)。Akt涉及腫瘤發生的構想證實，已經於臨床前期獲得驗證，顯示Akt之異位表現誘導惡性轉形與促成細胞存 9 200831525 活（Sun Μ.等人，Am J Pathol，2001· 159(2): ρ· 431-7; Cheng J.Q·等人，致癌基因，1997.14(23)·· p.2793-801);及Akt徑路的摧毁抑制細胞生長及誘導細胞凋亡(JetztA·等人，癌症研究，2003.63 (20): P.6697-706)。 5 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劑的臨床發展中斷。此外，受體酪胺酸激酶諸如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 10 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。如此，曲西立濱化合物或其衍生物與一種或多種鉑化合物之組合物具有展望可用作為腫瘤、癌症及異常細胞增 15 生之可能之組合治療。 L發明内容2 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鉑化合物之組合治療來於個體治療腫瘤或癌 20 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及相關化合物之胞毒性效應特別敏感，經由一種或多種鉑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鉑化合物藉以下一種方式或其組合 200831525 來治療腫瘤及癌症：⑴將曲西立濱及一種或多種鉑化合物投予對曲西立濱化合物或一種或多種鉑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/或一種或多種鉑化合物之毒性，同時仍然具有功 5 效；或(iii)使用所述用藥計劃，其可最小化曲西立潰化合物及/或一種或多種翻化合物之毒性。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：Akt is a serine/threonine protein kinase (also known as PKB), and Akt has three family members Aktl, Akt2 and Akt3. Stimulation of cells with growth factors or survival factors results in recruitment to the receptor for phosphokinase phosphoinositide-3-OH-20 kinase (PI3K), which phosphorylates phosphoinositide-4,5-diphosphate (PIP2) As PIP3, PIP3 recruits Akt to the serosa, in the serosal membrane, Akt by Thr308 and Ser473 (Aktl), Thr308 and Ser474 (Akt2) and Thr308 and Ser472 (Akt3) (Datta SR et al., Gene Dev, 1999·13 ( 22): p. 2905-27) is acidified and activated. Thus, PI3K activates Akt by converting acidification PIP2 200831525 " 5 r • and PIP2 into PIP3. The phosphatase PTEN dephosphorylates PIP3 to PIP2, thus preventing the activation of Akt. Most human cancers contain over-activated Akt (Datta SR et al., Gene Dev, 1999. 13(22): ρ·29〇5-27; BellacosaA· et al., International Journal of Cancer, 1995. 64(4): p .280-5 ; Sim Μ. 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 is over-expressed and/or highly activated (Roy HK et al., Cancer, 2002. 23 ( 1): ρ·201_5 ; Altomare DA et al., J Cell Biochem, 2003. 88(1): ρ· 10 470-6 ; Sun Μ· et al., Cancer Research, 2001· 61(16): P.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): ρ· 9267-71; 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 15, including the degree of overexpression of receptor tyrosine kinase and/or its ligands and deletion of phosphatase PTEN Loss (Khaleghpour K et al., Cancer Development' 2004. 25(2); ρ·241-8; Sekharam M· et al., Cancer Research, 2003, 63(22): ρ·7708·16; Cohen BD et al. Biochem Soc Symp, 1998.63: p.199-210; Muller WJ et al, Biochem Soc 20 Symp, 1998. 63: p. 149-57; Miller WE et al, J Virol, 1995 · 69(7): ρ·4390 -8; Slamoii Dj et al., Science, 1987. 235 (4785): p. 177-82; Andmlis. LL et al, j Clin Oncol, 1998. 16(4): p. 1340-9). The concept of Akt involved in tumorigenesis confirms that it has been validated in the preclinical phase, showing that ectopic manifestations of Akt induce malignant transformation and contribute to cell survival (Sun Μ. et al, Am J Pathol, 2001·159(2): ρ· 431-7; Cheng JQ· et al., Oncogene, 1997.14(23)·· p. 2793-801); and destruction of the Akt pathway inhibits cell growth and induces apoptosis (JetztA et al., Cancer Research) , 2003.63 (20): P.6697-706). 5 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 clinical disruption of a variety 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 for chemotherapy Drug-resistant and cancer patients have short survival times. Current research strategies emphasize the study of effective treatment modalities with lower risks. Thus, a combination of a trichostatin compound or a derivative thereof and one or more platinum compounds has a combination therapy that is expected to be useful as a tumor, cancer, and abnormal cell. SUMMARY OF THE INVENTION 2 SUMMARY OF THE INVENTION The present invention provides a combination therapy of triclinbine, triclinide and related compounds in combination with one or more platinum compounds to treat a tumor or cancer in an individual while limiting systemic toxicity. The present invention is based on the discovery that tumors or cancers that overexpress Akt kinase are particularly sensitive to the cytotoxic effects of TCN and related compounds, and synergistic effects can be obtained via a combination of one or more platinum compounds. Contrary to prior art and experience, the inventors have determined how to successfully use triclinidine and one or more platinum compounds to treat tumors and cancer in one of the following ways or a combination thereof: (1) to treat triclinide and one or more platinum The compound is administered to a patient having an increased sensitivity to the triclinic compound or one or more platinum compounds; (ii) using a prescribed dosage level that minimizes the toxicity of the tricinemide compound and/or one or more platinum compounds While still having a work efficiency; or (iii) using the drug use schedule, it minimizes the toxicity of the tromethamine compound and/or one or more compounds. In one aspect of the invention, the invention encompasses a composition comprising: (1) a compound of formula I-IV:

CHiCHi

9¾93⁄4

11 200831525 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）；酿基（包括低碳醯基）；烷基（包括低碳烷基）；醯胺、確酸醋包括烧基或芳基烷基；磺醯基包括甲磺醯基及节 · 5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、酿基；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R，、R3，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、烷基（包括低碳烷基）；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，知貝包括填脂質；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，_磷醚脂質或5，_醚脂質投藥；，11 200831525 wherein R2', R3, and r5 are respectively hydrogen, optionally substituted phosphate or phosphonate (including monophosphate, diphosphate, or triphosphate or stabilized phosphate prodrug); Carboalkyl); alkyl (including lower alkyl); decylamine, acid vinegar including alkyl or arylalkyl; sulfonyl including methylsulfonyl and benzyl group, wherein the phenyl is as needed Substituted by one or more substituents as exemplified in the definition of aryl group as described herein; optionally substituted aryl sulfonate, brewing group; lipids including phospholipids; amino acids; carbohydrates; a peptide; or, cholesterol; or other pharmaceutically acceptable leaving group, which is available in vivo for a compound wherein R, R3, or r5 are hydrazine or monophosphate, diphosphoric acid, Or triphosphate; wherein Rx and Ry are respectively hydrogen, optionally substituted phosphate; sulfhydryl (including low carbon fluorenyl); decylamine, alkyl (including lower alkyl); aromatic, deoxygenated An alkyl group such as polyethylene glycol, an optionally substituted arylsulfonyl group, which includes a fat-filling Amino acid; carbohydrate; peptide; or cholestrol; or other pharmaceutically acceptable leaving group; in one embodiment, - the compound is a 5, _phosphonate lipid 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、鲁或％狀烷基（包括低碳烷基）、烯基或炔基、c〇_烷基、c〇_ 歸基、CO·炔基、CO-芳基或雜芳基、co_烧氧基烷基、c〇_ 20芳氧基烷基、c〇-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； (ii)如式V之一種或多種鈾化合物： 12 200831525 I白化合物Ri and R2 are each a linear, branched, ru or hydroxy-type alkyl group (including a lower alkyl group), an alkenyl group or an alkynyl group, a c〇-alkyl group, and a c〇_ group, respectively, which may be substituted. , CO. alkynyl, CO-aryl or heteroaryl, co-alkoxyalkyl, c〇 20 aryloxyalkyl, c〇-substituted aryl 'sulfonyl, alkyl sulfonium a arylsulfonyl group, an arylalkylsulfonyl group; (ii) a uranium compound of the formula V: 12 200831525 I white compound

式v 其中： 5 R!、R2、R3及I各自分別為氫、視需要可經取代之胺、Wherein: 5 R!, R2, R3 and I are each hydrogen, optionally substituted amines,

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、Ri、R2、R3及R4間所形成之芳香環、雜芳香環或環狀環； R5及R6各自分別為鹵素、視需要可經取代之烧氧基、 10 視需要可經取代之酯、視需要可經取代之烷基胺、芳香族胺、雜芳香族胺； (iii)藥學上可接受之載劑。於另一個實施例中，本發明涵蓋一種於一哺乳動物治療腫瘤或癌症之方法，包括對該哺乳動物投予有效量之一 15 種組成物包括： ⑴式I-IV化合物：宇 Ηΐ ΟΜιAn aromatic amine, a heteroaromatic amine, an optionally substituted straight chain, branched or cyclic alkyl group, an aromatic ring, a heteroaromatic ring or a cyclic ring formed between Ri, R2, R3 and R4; R5 and R6 Each is a halogen, an optionally substituted alkoxy group, 10 optionally substituted esters, optionally substituted alkylamines, aromatic amines, heteroaromatic amines; (iii) pharmaceutically acceptable Carrier. In another embodiment, the invention encompasses a method of treating a tumor or cancer in a mammal comprising administering to the mammal an effective amount of one of 15 compositions comprising: (1) a compound of formula I-IV: 宇 Ηΐ ΟΜι

13 20083152513 200831525

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（包括一麟酸、二構酸、或三鱗酸或安定化構酸 5 前藥）；醯基（包括低碳醯基）；烧基（包括低碳烧基）；醯胺、石黃酸酯包括烧基或芳基烧基；績蕴基包括曱石黃醢基及节基，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'或三鱗酸；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$炭驢基）；醯胺、烧基（包括低碳烧基）；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； 14 200831525Wherein R2', R3' and R5' are respectively hydrogen, optionally substituted phosphate or phosphonate (including a linonic acid, a dibasic acid, or a tribasic acid or a hydrating acid 5 prodrug); (including low carbon sulfhydryl groups); alkyl groups (including low carbon alkyl groups); decylamines, retinyl esters including alkyl or aryl alkyl groups; the base includes fluorite and sulfhydryl groups, wherein the phenyl group Desirable may be substituted with one or more substituents as described in the definition of aryl as described herein; optionally substituted arylsulfonyl; lipids including phospholipids; amino acids; carbohydrates; a peptide; or 10 cholesterol; or other pharmaceutically acceptable leaving group which provides a compound in vivo wherein R2', R3' or R5' are hydrazine or monophosphate, diphosphate or trisquaric acid, respectively; Rx and Ry are respectively hydrogen, optionally substituted phosphate; sulfhydryl (including lower than anthracenyl); decylamine, alkyl (including low carbon alkyl); aromatic, 15 deoxyalkylene such as Polyethylene glycol, optionally substituted arylsulfonyl; lipids including phospholipids; amino acids; Carbohydrate; peptide; or cholesterol; or other pharmaceutically acceptable leaving group; in one embodiment, the phosphorus compound is the 5'-ether lipid, or as a 5'-ether lipid administration; 14200831525

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、CO-烷基、CO-稀基、CO-炔基、CO-芳基或雜芳基、c0_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、芳基石買酿基、芳烧基確醯基；以及 (11)如式V之一種或多種鉑化合物：鉑化合物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 CO-alkyl group, a CO-saturated group, or a CO- group which may be substituted as needed. Alkynyl, CO-aryl or heteroaryl, c0_alkoxyalkyl, co-aryloxyalkyl, CO-substituted aryl, sulfonyl, alkylsulfonyl, aryl stone a aryl group; and (11) one or more platinum compounds of the formula V: a platinum compound

式V 10 其中：Formula V 10 where:

Ri、R2、R3及R4各自分別為氫、視需要可經取代之胺、芳香無胺、雜芳香族胺、視需要可經取代之直鏈、分支或 %狀院基、Rl、R2、R3及R4間所形成之芳香環、雜芳香環或環狀環；尺5及R6各自分別為鹵素、視需要可經取代之烧氧基、視而要可經取代之酯、視需要可經取代之烷基胺、芳香族 fee、雜芳香族胺。忒等方法可用於治療對TCN、TCN-p及/或相關化合物 2〇 =毒性效應特別敏感之腫瘤及癌症。於另一個實施例中， ;甫乳動物特別為人類治療腫瘤之方法包括⑴由腫瘤獲传生物樣本；⑼判定軸瘤是㈣度表現Akt激酶；以 ()乂如此處所述之曲西立濱、磷酸曲西立濱或相關化合 15 200831525 物組合-種或多種鈾化合物來治療過度表現施激酶之腫瘤於另個貝施例中’备由檢定分析腫瘤或癌症是否有雜化施激酶的存在，例如經由使用可檢_酸化形式之抗體來判定Akt激酶之表現程度。於另—個實施例中，經纟 - 5才欢疋刀析得自個體之腫瘤細胞或癌細胞且比較該含量與對 - 照組織之含量，可判定Akt之過度表郷度。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磷酸化之Akt激酶。馨 10 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、血中澱粉酶過 15高、腹瀉、胃炎及/或發燒。於另一個實施例中，TCN、TCN-P * 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%反應或完全反修應0 於另一個實施例中，提供一種方法來治療已經診斷為 20患有腫瘤之個體，係經由對該個體投予有效量之TCN、 TCN-P或相關化合物及一種或多種鉑化合物，該投藥係根據一種給藥計劃，包括約略每週一次投予曲西立濱化合物及一種或多種鉑化合物約經3週，接著為未投予曲西立濱化合物及一種或多種鉑化合物之一週時間。另一個實施例 16 200831525 10 中’提供-種於-個體治療腫瘤或癌症之方法，係經由對該個體投予-種給藥計劃，每週各—次㈣碱克/平方米或以下之TCN、TCN-P或才目關化合物及—種或多種翻化合物。於另-個實施例中，曲西立濱化合物及一種或多種翻化合物可以短時間例如約5、1〇或15分鐘時間以單一大劑里投予。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-種或多種鉬化合物係透過連續輸注經歷至少24、48、72、96或12()小時時間奸。於若干實施例中，連續投藥可每週至少重複—次、每兩週重複一次、及/或每個月重複-次。於其它實施例中，曲西立濱化合物及一種或多種鉑化合物可每三週至少投予—次。於額外實施例中，化合物可每曰至少一次投藥經歷至少2、3、4或5 曰。於名員外實施例中，如此處揭示之曲西立濱化合物及一 15種或多種鉑化合物可以有效造成腫瘤退行之數量投予病人。曲西立濱化合物及一種或多種鉑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 20 如此處揭示之一種或多種鉑化合物可投予一個體。曲西立濱化合物及一種或多種鉑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20毫克/平方米曲西立濱化合物及一種或多種鉑化合物。於一個實施例中，每週一次投予低於10毫克/平方米 17 200831525 曲西立濱化合物及少於約ίο毫克/千克之一種或多種鉑化合物。於額外實施例中，低於2毫克/平方米、5毫克/平方米、 10毫克/平方米、15毫克/平方米、20毫克/平方米、25毫克/ 平方米、35毫克/平方米、45毫克/平方米、55毫克/平方米、 5 65毫克/平方米、75毫克/平方米、85毫克/平方米、及/或95 毫克/平方米劑量之曲西立濱化合物及低於2毫克/平方米、5 毫克/平方米、10毫克/平方米、15毫克/平方米、20毫克/平方米、25毫克/平方米、35毫克/平方米、45毫克/平方米、 55毫克/平方米、65毫克/平方米、75毫克/平方米、85毫克/ 10 平方米、及/或95毫克/平方米劑量之一種或多種鉑化合物投予一個體。於另一個實施例中，低於100毫克/平方米曲西立濱化合物可透過連續輸注經歷至少5日投予一個體。於特定實施例中，如此處揭示之曲西立濱化合物及一種或多種鉑化合物可用於胰癌、攝護腺癌、大腸直腸癌及/或卵巢癌 15 之治療。於另一個實施例中，曲西立濱化合物及一種或多種鉑化合物及/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/或骨髓瘤。於本發明之其它實施例中，曲西立濱化合物及一種或多種鉑化合物可用於治療實體腫 20 瘤。又有其它實施例中，此處揭示之曲西立濱化合物及一種或多種鉑化合物及組成物可用於治療腫瘤或癌症，諸如但非限於下列器官或組織之癌症：***、攝護腺、硬骨、肺、大腸包括但非限制大腸直腸、尿路、膀胱、非何杰金氏淋巴瘤、黑素瘤、腎臟、腎上腺、胰臟、咽頭、曱狀腺、 200831525 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鉑化合物可用於胰癌、乳癌、大腸直腸癌、及/或卵巢癌之治療。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鉑化合物可用於治療血管新生 . 5相關疼病。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 … 多種鋼化合物來治療白血病。於其它實施例中，連續輸注例如町重複每兩週、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 1〇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鉑化合物視需要可組合藥學上可接受之載劑。於一個實施例中，曲西立濱化合物及一種或多種鉑化合物及組成物可組合投予，形成該組成物之一部分，或可 ' 15同時或不同時呈分開組成物投予。於其它貫施例中，如此處揭示之曲西立濱化合物及一鲁種或多種顧化合物可用來治療對-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 2〇化合物及一種或多種鉑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腫瘤或癌症之病人之有效量投予’包括單獨對紫杉醇（tax〇l)、拉帕黴素 (rapamycm)、塔莫西芬(tamoxifen)、西鉑汀㈣叩以㈤及/或傑費堤尼（gefitimb)(艾瑞莎（iressa))有抗藥性之腫瘤或癌 19 200831525 症。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鉑化合物；或以可有效於宿主治療腫瘤、癌症及其它與異常 5 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顧化合物之藥學組成物。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S旨投予有需要之個體，其中該癌症例如為癌瘤、 10 肉瘤、淋巴瘤、白血病、或骨髓瘤。該化合物或其鹽、異構物、前藥或酯視需要可呈藥學上可接受之組成物提供，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 15 投予。本發明也包括如此處揭示之化合物或其鹽、前藥或酯用於治療腫瘤或癌症之用途，視需要可於藥學上可接受之載劑；以及如此處揭示之曲西立濱化合物及一種或多種鉑化合物或其鹽、前藥或酯視需要可於藥學上可接受之載 20 劑，用於製造癌症或腫瘤治療用藥之用途。，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。A顯示API-2 (曲西立濱）之化學結構式。B顯示API-2抑制於AKT2轉形NIH3T3細胞中之 200831525 AKT2辑度。a_(Wile)型Am轉顧腿η細胞以 API 2 (ΙμΜ)處理I曰不之時間，且接受以抗石舞酸旭_τ獅 _ 及_8473抗體（上®及巾圖）之免疫墨點分析。下圖顯示總 ΑΚΤ2之表現° C顯不αρι_2抑制Akt之三個同質異形體。 :5 HEK293 細胞於 EGF 刺激前以 HA-Aktl、HA_AKT2、及 HA-AKT3轉移感染且以Αρι·2 (1_)或瓦特曼寧 “ (wortmannin) (15μΜ)處理，細胞經溶解且以anti HA抗體免疫沉澱。免疫沉殿物接受試管内激酶檢定分析（上)及以抗碟鲁酸_Akt_T3G8⑺抗體進行免疫墨點分析。巾圖顯示經轉移 10感染之Aktl、AKT2、及aKT3之表現。D顯示Αρι·2於試管内不會抑制Akt。於試管内組成活性八1^丁2重組蛋白質於激酶緩衝液之激酶檢定分析含有ΙμΜ API-2 (線道3)。第2圖驗證ΑΠ-2不會抑制PI3K、PDK1及AGC激酶家族中之緊德相關成員。A顯示試管内ρΐ3Κ激酶檢定分析。於 : 15 EGF刺激前，HEK293細胞經血清匱乏且以API-2 (1 _)或瓦特曼寧（15μΜ)處理30分鐘。細胞經溶解且以anti-pll〇〇；抗 ® 體免疫沉澱。免疫沉澱物使用PI-4-P作為酶基質接受活體内激酶檢定分析。B顯示API-2對試管内PDK1活化之效應（上圖），實心圓顯示藉API-2抑制。空心圓顯示藉陽性對照史 20 妥洛史寶靈(staurosporine)抑制，史妥洛史寶靈為強力pDKi 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-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。c 顯示PKCa以抗磷酸-PKCa _T638(上圖）及總PKCa (下圖） 21 200831525 抗體接著以API-2或非選擇性PKC抑制劑Ro3丨_8220處理之石粦酸化程度之免疫墨點分析。D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 5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。E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 Biotechnology Inc))中培養。定量激酶活性。於ρ中 10 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 (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凋亡。A為使用API-2處理後 15 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）。於B，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 (CellTiter) 96細胞增生檢定分析套件組（普米嘉公司 20 (Promega))分析。C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。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A，驗證API-2可抑制結節素(tuberin)、Bad、AFX及GSK-3 200831525 /3之Akt磷酸化。於以ΑΠ-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。Β顯示Αρι_2可抑制腫瘤生長。腫瘤細胞注射入裸小鼠體内，左侧Akt細胞濃度低，而右側Akt細胞濃度高。當腫瘤達到約立方毫 5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10個腫瘤之平均值。c顯示帶有以API-2 或載媒劑（對照組)處理之OVCAR3 (右)及OVCAR5 (左）之小鼠之代表圖。D顯示於實驗結束時之腫瘤大小（下）及重量（上) 之實例。E中，腫瘤溶解產物之免疫墨點分析係使用經以 10 ΑΠ-2處理(T3及T4)及未經處理(T1及T2)之〇 VC AR-3 -衍生腫瘤中之抗填-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PDK1與Akt之重組株於含磷酸基肌糖醇-3,4,5-P3 (PIP3)、ΑΠ-2及組織呋H2B作 15 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。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 20 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。Ri, R2, R3 and R4 are each independently hydrogen, optionally substituted amine, aromatic amine-free, heteroaromatic amine, optionally substituted linear, branched or %-like, Rl, R2, R3 And an aromatic ring, a heteroaromatic ring or a cyclic ring formed between R4; each of the feet 5 and R6 is a halogen, an alkoxy group which may be substituted as needed, an ester which may be substituted, and may be substituted as needed Alkylamine, aromatic fee, heteroaromatic amine. Methods such as sputum can be used to treat tumors and cancers that are particularly sensitive to TCN, TCN-p and/or related compounds 2 〇 toxic effects. In another embodiment, the method for treating a tumor, particularly for a human, comprises (1) obtaining a biological sample from the tumor; (9) determining that the axon is a (four) degree of Akt kinase; (), as described herein, Qu Cixi Parabens, Tricineribin phosphate or related compounds 15 200831525 Combination of species or multiple uranium compounds to treat tumors that overexpressed the application of kinases in another benfactory example - to determine whether a tumor or cancer has hybridized kinases by assay There is, for example, the extent to which Akt kinase is expressed by the use of antibodies in a detectable-acidified form. In another embodiment, the tumor cells or cancer cells obtained from the individual are analyzed by 纟-5, and the content of the content and the content of the tissue are compared to determine the excessive expression of the Akt. In several embodiments, Akt can be at least 2, 2.5, 3, or 5 times overexpressed in a comparison of cancer samples to a control group. In several embodiments, the overexpressed Akt kinase can be an overactive and phosphorylated Akt kinase. Xin 10 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 reduces or eliminates toxic side effects including, but not limited to, hepatotoxicity, thrombocytopenia, hyperglycemia, vomiting, hypocalcemia, anemia, hypoalbuminemia, myelosuppression, blood Triglyceride is too high, blood amylase is 15 high, diarrhea, gastritis and/or fever. In another embodiment, the administration of TCN, TCN-P* or related compounds can provide at least a portion, such as at least 15%, 20% or 30%, in vivo or 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 20 having a tumor by administering to the individual an effective amount of TCN, TCN-P or related compound and one or more platinum compounds, The administration is based on a dosing schedule comprising administering the triclinibine compound and one or more platinum compounds approximately once a week for about 3 weeks, followed by one week of not administering the tricilide compound and one or more platinum compounds. time. Another embodiment 16 200831525 10 'providing - a method for treating a tumor or cancer in an individual, by administering a dose to the individual, a weekly dose of TCN for each (four) base grams per square meter or less TCN-P or the compound and one or more compounds. In yet another embodiment, the triclinide compound and one or more compounds can be administered in a single bolus for a short period of time, e.g., about 5, 1 or 15 minutes. In still other embodiments, a dosing schedule is provided wherein the triclinide compound and the one or more molybdenum compounds are administered through a continuous infusion for at least 24, 48, 72, 96 or 12 () hours. In several embodiments, continuous administration can be repeated at least once a week, repeated every two weeks, and/or repeated every other month. In other embodiments, the triclinide compound and one or more platinum compounds can be administered at least once every three weeks. In additional embodiments, the compound may be administered at least 2, 3, 4 or 5 Torr per at least one administration. In the examples outside the celebrity, the tromethamine compound and one or more platinum compounds as disclosed herein can be administered to a patient in an amount effective to cause tumor regression. Administration of the triclinide compound and one or more platinum 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 subject in vivo. In some embodiments, at least 2, 5, 10, 15, 20, 30 or 50 mg/m2 of the triclinide compound and 20 one or more of the platinum compounds as disclosed herein can be administered to one body. Administration of the tripholidine compound and one or more platinum 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 20 mg/m2 of the triclinbine compound and one or more platinum compounds. In one embodiment, less than 10 mg/m 2 17 200831525 trichostatin compound and less than about ί mg/kg of one or more platinum compounds are administered once a week. In additional embodiments, less than 2 mg/m 2 , 5 mg/m 2 , 10 mg/m 2 , 15 mg/m 2 , 20 mg/m 2 , 25 mg/m 2 , 35 mg/m 2 , 45 mg/m2, 55 mg/m2, 5 65 mg/m2, 75 mg/m2, 85 mg/m2, and/or 95 mg/m2 dose of trichostatin compound and less than 2 Mg/m2, 5 mg/m2, 10 mg/m2, 15 mg/m2, 20 mg/m2, 25 mg/m2, 35 mg/m2, 45 mg/m2, 55 mg/ One or more platinum compounds of a square meter, 65 mg/m 2 , 75 mg/m 2 , 85 mg / 10 m 2 , and/or 95 mg/m 2 dose are administered to one body. In another embodiment, less than 100 mg/m2 of the triclopibine compound can be administered to a body for at least 5 days through continuous infusion. In a particular embodiment, the triclinide compound and one or more platinum compounds as disclosed herein are useful in the treatment of pancreatic cancer, prostate cancer, colorectal cancer, and/or ovarian cancer. In another embodiment, the triclinide compound and one or more platinum compounds and/or the therapeutic plan of the invention can be used to treat cancer, sarcoma, lymphoma, leukemia and/or myeloma. In other embodiments of the invention, the triclinide compound and one or more platinum compounds are useful for treating a solid tumor. In still other embodiments, the tromethamine compound disclosed herein and one or more platinum compounds and compositions are useful for treating a tumor or cancer, such as, but not limited to, cancer of the following organs or tissues: breast, prostate, bony , lung, large intestine including but not limited to the large intestine rectum, urinary tract, bladder, non-Hodgkin's lymphoma, melanoma, kidney, adrenal gland, pancreas, pharynx, sacral gland, 200831525 stomach, brain and / or ovary. In a particular embodiment, the triclinide compound and one or more platinum compounds are useful in the treatment of pancreatic cancer, breast cancer, colorectal cancer, and/or ovarian cancer. In other embodiments of the invention, the tromethamine compound disclosed herein and one or more platinum compounds are useful in the treatment of angiogenesis. In several embodiments, the treatment of leukemia via continuous infusion of the triclinide compound and one or more steel compounds is provided by continuous infusion for at least 24, 48, 72 or 96 hours. In other embodiments, a continuous infusion, such as a town, is repeated at least once every two weeks, every three weeks, or every four weeks. • In a specific embodiment, a method of treating a tumor and other disorders associated with abnormal cell proliferation in a host, the method comprising administering to the host an effective amount of a triclinide compound and one or more platinum compounds A pharmaceutically acceptable carrier can be combined as needed. In one embodiment, the triclinide compound and one or more platinum 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 tromethamine compound as disclosed herein and a genus or a plurality of compounds can be used to treat tumors or cancers that are resistant to one or more known anticancer drugs, including those disclosed herein. Examples of tumors or cancers and compounds. In one embodiment, an effective amount of a tromethamine 2 oxime compound and one or more platinum compounds as disclosed herein for treating a patient having a drug resistant tumor or cancer, such as a multi-drug resistant tumor or cancer, is disclosed. Invested 'includes paclitaxel alone, rapamycm, tamoxifen, xiplatin (four) ( (5) and/or gefitimb (area) (iressa)) A drug-resistant tumor or cancer 19 200831525. In several embodiments, a method is provided comprising administering to a host in need thereof an effective amount of a triclinide compound and one or more platinum compounds as disclosed herein; or effective treatment of a tumor, cancer, and other abnormalities with the host 5 A therapeutically effective amount of a condition associated with cell proliferation includes a citrilide compound and one or more pharmaceutical compositions of the compound. 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 S thereof, to a subject in need thereof, wherein the cancer is For example, cancer, 10 sarcoma, lymphoma, leukemia, or myeloma. The compound or a salt, isomer, prodrug or ester thereof may optionally be provided as a pharmaceutically acceptable composition, and the composition comprises a suitable carrier such as water, which is formulated for administration to a patient in need thereof. use. Optionally, the compound can be administered in combination with an additional therapeutic agent of at least one tumor or cancer, or alternately administered with the additional therapeutic agent. The invention also includes the use of a compound as disclosed herein, or a salt, prodrug or ester thereof, for the treatment of a tumor or cancer, a pharmaceutically acceptable carrier, if desired; and a tromethamine compound as disclosed herein and a Or a plurality of platinum compounds or salts, prodrugs or esters thereof, if necessary, can be used in the manufacture of a cancer or a tumor therapeutic drug in a pharmaceutically acceptable carrier. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 shows the identification of API-2 (Quxiribin) identified as a candidate for Akt inhibitors in the NCI diversity set. A shows the chemical structure of API-2 (Quxi Libin). B shows that API-2 is inhibited in the 200831525 AKT2 series in AKT2 transformed NIH3T3 cells. A_(Wile) type Am refers to the leg η cells treated with API 2 (ΙμΜ) for a period of time, and receives immunoblotting with anti-Shi Wuxu _ _ _ _ _ and _8473 antibodies (upper and towel map) Point analysis. The figure below shows the performance of total ΑΚΤ2 ° C 不αρι_2 inhibits three homomorphisms of Akt. :5 HEK293 cells were infected with HA-Akt1, HA_AKT2, and HA-AKT3 before EGF stimulation and treated with Αρι·2 (1_) or wortmannin (15μΜ), cells were lysed and anti-anti-H antibody Immunoprecipitation. Immunosuppressive substances were subjected to in vitro kinase assay (top) and immunoblot analysis was performed with anti-luruic acid _Akt_T3G8(7) antibody. The towel map showed the performance of Aktl, AKT2, and aKT3 infected by metastasis. Αρι·2 does not inhibit Akt in the test tube. The kinase assay for the active octapeptide 2 in the test tube contains the ΙμΜ API-2 (line 3) in the test tube. Figure 2 Verify ΑΠ-2 No It inhibits the related members of the PI3K, PDK1, and AGC kinase families. A shows an in vitro assay for ρΐ3Κ kinase assay. After: 15 EGF stimulation, HEK293 cells are serum-deficient and API-2 (1 _) or Watmanning (15 μΜ) for 30 minutes. The cells were lysed and immunoprecipitated with anti-pll〇〇; anti-body. Immunoprecipitates were assayed for in vivo kinase assay using PI-4-P as the enzyme matrix. B shows API-2 test The effect of PDK1 activation in the tube (above) The circle shows inhibition by API-2. The open circle shows inhibition by the positive control history of 20 staurosporine, and Stallosporin is a potent pDKi 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. Immunoblots were detected with the indicated antibodies. c shows PKCa with anti-phospho-PKCa _T638 (top panel) and total PKCa (bottom) 21 200831525 The antibody was then analyzed by immunoblotting of the degree of osmotic acidification treated with API-2 or the non-selective PKC inhibitor Ro3丨_8220. D shows in vitro SGK kinase assay. HEK293 cells were stimulated before EGF Infected with HA-SGK and treated with API-2 or Watmaning. The kinase of the in vitro test was immunized with ha-SGK 5 precipitation using MBP as the enzyme substrate (top panel). The figure below shows HA-SGK transfected with infection. Performance. E shows the results of PKA kinase assay. Immunopurified PKA in ADB buffer containing the appropriate inhibitor (API_2 or PKAI) and enzyme matrix Kemptide (Upstate Biotechnology) Inc)) Quantitative kinase activity. Western blots were shown in ρ 10 . OVCAR3 cells were treated with API-2 for the indicated time. The cell lysate is immunized with the appropriate anti-phospho antibody (1 - 4 map) and anti-actin antibody (bottom panel). Figure 3 demonstrates that API-2 induces apoptosis in Akt-expressing human cancer cells by inhibiting Akt activity and cell growth. A is a Western blot after treatment with API-2. The degree of phosphorylation of Akt is detected by an anti-phospho-Akt-T308 antibody in a suitable human cancer cell line. The ink spots were again probed with anti-total Akt antibodies (bottom panel). At B, a cell proliferation assay was performed. 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 CellTiter 96 Cell Proliferation Assay Kit (Promega). C provides apoptosis analysis. Cells were treated with API_2, stained with annexin V and PI, and analyzed by FACScan. Figure 4 shows in a mouse xenograft sheet. In a cancer cell line with elevated Akt, API-2 inhibits downstream targets of Akt and has antitumor activity. At A, validation of API-2 inhibited Akt phosphorylation of tuberin, Bad, AFX and GSK-3 200831525 /3. After treatment with ΑΠ-2, OVCAR3 cells were lysed and subjected to immunoblot analysis with applicable antibodies. Β shows that Αρι_2 can inhibit 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 cubic millimeters, the animal was treated with vehicle or at 1 mg/kg/day API-2. Each measurement represents the average of 10 tumors. c shows a representative of mice with OVCAR3 (right) and OVCAR5 (left) treated with API-2 or vehicle (control). D shows examples of tumor size (bottom) and weight (top) at the end of the experiment. In E, the immunoblot analysis of tumor lysates was performed using anti-fill-Akt- in VC AR-3-derived tumors treated with 10 ΑΠ-2 (T3 and T4) and untreated (T1 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 was performed using a recombinant strain of PDK1 and Akt in a kinase buffer containing phospho-inositol-3,4,5-P3 (PIP3), guanidine-2 and tissue furose H2B as the enzyme substrate. After 30 minutes of 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. Figures 7a-7d provide mRNA and amino acid sequences of human Akt2, also indicating the position of the 20-cutase. Figures 8a-8c provide mRNA and amino acid sequences of human Akt3, also indicating the position of the restriction enzyme.

第9圖顯示西顧汀（cisplatin) (CDDP)抗性細胞系内生性表現超活性Akt。第9A圖顯示兩種卵巢癌細胞系（A2780S 23 200831525 及OV2008)及以遞增量西鉑汀處理之其西鉑汀抗性對偶（分別為A2780CP及C13)之線圖。西舶汀抗性變異株亦即 A2780CP及C13驗證回應於西鉑汀之存活率增高。第9β圖暴員 5 示西鉑汀敏感性卵巢癌細胞亦即A2780S及OV2008與其西鈿汀抗性對偶之比較。上圖顯示於西鉑汀抗性細胞系及A2780S中之超活性Akt。下圖顯示於各類細胞中存在之 Akt總量。第10圖顯示曲西立濱（TCN) (ΑΠ-2)可克服西翻汁抗 _ 性。西鉑汀抗性細胞系亦即A2780S及C13係以〇、5、1〇及馨 10 20μΜ西鉑汀處理。白色條紋柱表示只以西鉑汀處理。黑灰柱表示以西鉑汀與10μΜΊ^Ν處理。以TCN共同處理可降低細胞存活率，指示TCN可克服西鉑汀抗性。第11圖顯示使用T C N及西鉑汀對C丨3卵巢癌細胞之細胞存活率所見協同效應。C13細胞以…卜卜1〇&2(^mtcn - 15含及未含ΐ〇μΜ西齡處理。西財與Τ(：Ν之組合物可協@ 性地降低細胞存活率。，第I2圖顯示TCN可提升西財抑制於裸小鼠異種移植 ^ 片中C13印巢癌細胞生長之能力。裸小鼠接受⑶異種移植片，接著以載媒劑(DMSO)、單獨西如丁、單獨TCN或西鉑㈣TCN處理’接著分析於接種後7、i4、21、或時之腫瘤退行。左圖（A)顯示一線圖驗證西銘汀與tcn之組合物顯著㈣腫瘤之生長。右_)顯示於接受代表性處理之接種小鼠體内之腫瘤質量。第圖，員不於人印巢癌細胞中他徑路藉抑制劑 24 200831525 之快速活化。上圖（A)顯示於使用1 nM mTOR抑制劑拉帕黴素(rapamycin)處理指示之時間後，OV3細胞溶解產物之西方墨點分析。膜係以對磷酸化同基因型及對總蛋白質具有 ^ 特異性之抗體探測用於Akt、p70S6K及FKHRL 1之比較。下 - 5 圖（B)顯示於使用mTOR抑制劑HAD001處理指定之時間後，MCF7細胞溶解產物之西方墨點分析。膜係以對磷酸化 … 同基因型及對總蛋白質具有特異性之抗體探測用於Akt及 p70S6K之比較。 • 第14圖顯示透過mTOR抑制劑AKT徑路之活化係藉 10 TCN衰減。左圖顯示於〇V3細胞中TCN對於以拉帕黴素處理之Akt磷酸化之效應。右圖（B)顯示於MCF7細胞中TCN對以RAD001處理之Akt磷酸化之相同效應。第15圖顯示經由TCN與RAD001或拉帕黴素之組合對細胞生長之效應提升。左上圖（A)為線圖，顯示以對照組、 * 15 單獨拉帕黴素、單獨TCN或TCN與拉帕黴素處理6曰， DU-145細胞之細胞數目。右上圖（B)為線圖，顯示以對照 • 組、單獨RAD001、單獨TCN或TCN與RAD001處理6日， MCF7細胞之細胞數目。下圖（〇為線圖，顯示以對照組、單獨拉帕黴素、單獨TCN或TCN與拉帕黴素處理6日，OV3 20 細胞之細胞數目。第16圖顯示經由奥羅拉(Aurora)-A活化Akt誘導細胞存活及化學抗性。上圖（A)顯示於A2780S、A2780CP及OV2008 卵巢癌細胞系中奥羅拉-A成功地轉移感染及表現。第二圖 (B)為線圖顯示於A2780S細胞中表現奥羅拉-A提高細胞存 25 200831525 活率及對太平洋紫杉醇(Paclitaxel)之抗性。第三圖（C)為線圖顯示於A2780S細胞中表現奥羅拉-A提高細胞存活率及對西鉑汀之抗性。下圖（D)顯示於〇¥2〇〇8細胞中奉現奥羅拉-A提高細胞存活率及對西鉑汀之抗性。 · 5 第17圖顯示奥羅拉-A對胞色素c釋放及Akt活化之效 - 應。第17A圖顯示於使用空白載體或使用或未使用西鉑汀處理之奥羅拉-A轉移感染之A2780S細胞中胞色素c之免疫螢：光。西鈾汀誘導胞色素c的釋放受奥羅拉—A表現之抑制。第 17B圖為西方墨點顯示奥羅拉_A之表現可提高於西鉑、汀敏鲁 10感細胞中之Akt之磷酸化。第17C圖顯示提高奥羅拉-a濃度可增加Akt之磷酸化及活化。第18圖顯示TCN(API-2)對奥羅拉_八誘導西鉑汀抗性之效應。上二圖（A及B)顯示以空白載體或奥羅拉_A轉移感染，接著單獨使用西鉑汀、單獨API-2或API-2及西鉑、;丁處 15理之A2780S細胞。下二圖（C及D)顯示以空白載體或奥羅拉 * -A轉移感染，接著單獨使用西鉑汀、單獨API-2或API-2及西麵、/丁處理之OV2〇〇8細胞。TCN降低於八278〇§及〇乂2〇〇8 兩種細胞中經由奥羅拉-A表現所提供之西鉑、汀抗性。第19圖顯示API-2、奥羅拉_A、p53及Akt於細胞存活所 20 扮演之角色之示意圖。第20圖顯示Src家族酪胺酸激酶抑制劑雷沙堤尼 (dasatinib)(史普利索（Sprycel))對八375細胞中之§!^麟酸化及Akt磷酸化之效應。Src酪胺酸激酶之抑制可提高Akt之鱗酸化。 26 200831525 • 5 【實施方式3 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翻化合物藉以下一種方式或其組合來治療腫瘤及癌症：⑴將曲西立濱及一種或多種鉑化合物只投予根據後述診斷試驗對曲西立濱化合物及/或一種或多種鉑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/或一種或多 • 種始化合物之毒性，同時仍然具有功效；或(iii)使用所述用 10 藥計劃，其可最小化曲西立濱化合物及/或一種或多種鉑化合物之毒性。 5.1定義如此處使用，「本發明化合物」一詞係指涵蓋式I-IV、式V化合物、結構式A-Ι化合物及其組合。 ^ 15 如此處使用，「癌症」及/或「癌性」等詞述及或說明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、 20 大腸癌、鱗狀細胞癌、小細胞肺癌、非小細胞肺癌、胃腸癌、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、腎癌及甲狀腺癌。其它癌症之非限制性實例為基底細胞癌；膽管癌；骨癌，腦及中搞神經糸統(CNS)癌；脈絡膜癌；結締組織癌； 27 200831525 食道癌：眼癌；頭頸癌；胃癌；上皮内腫瘤；咽癌；琳巴瘤包括何杰金氏淋巴瘤及非何杰金氏淋巴瘤；黑素瘤；骨髓瘤；神經母細胞瘤；口腔癌（例如唇、壬、π B备_s、. ± 方、口及喉頭），胰癌；視網膜母細胞瘤；橫紋肌肉瘤；直腸癌；呼吸系統癌； 5肉瘤；皮膚癌；胃癌；睪丸癌，·子宮癌；泌尿系統癌及其它癌瘤及肉瘤。如此處使用，「腫瘤」一詞係指全部贅生性細胞生長及增生，包括惡性或良性及全部癌前及癌性細胞及組織。例如’特定癌症可以實體團塊腫瘤為特徵。實體腫瘤團塊當 10存在時可為原發性腫瘤團塊。原發性腫瘤團塊係指由該組織之正常細胞轉形所導致於該組織内之癌細胞生長。於大部分情況下，原發性腫瘤團塊係以囊腫的存在來識別，可透過視覺方法或觸診方法來發現囊腫的存在，或藉由組織形狀、紋理或重量的不規則來發現囊腫。但若干原發性腫 15瘤無法觸診，只能經由醫療成像技術諸如X光(例如***攝影）或藉針刺抽吸來檢測。使用後述技術較為常見於早期檢測。組織内部之癌細胞的分子分析及表現型分析通常可驗證癌症是否為組織所内生的癌症，或病灶是否係來自於另一個部位的癌症轉移。 20 如此處使用，除非另行指示，否則烷基一詞包括例如為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、 200831525 2,2-二甲基丁基、及2,3_二甲基丁基。烷基視需要例如可經、乂個或夕個取代基取代，取代基諸如鹵原子(ρ、ci、Br 或 I)(例如 cf3、2泰乙基、ch2F、CH2a、CH2CF3、或 CF2CF3)、每基（例如CH2〇H)、胺基（例如CH2NH2、 5 ch2nhch3 '或CH2N(CH3)2)、烧基胺基、芳基胺基、烧氧基、芳氧基、硝基、疊氮基(例如CH2N3)、氰基(例WCH2CN)、礦酸、硫酸根、膦酸、磷酸根、或膦酸根，或為未經保護，或如热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， 10紐約之教示，以引用方式併入此處。除非另行陳明，否則如此處使用，低碳烷基一詞係指 (^至口飽和直鏈、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一個或兩個烧基 15 或芳基取代基之胺基。胺基酸一詞包括天然及合成α、/5、r或請基酸，包括但非限於蛋白質中出現之胺基酸，亦即甘胺酸、丙胺酸、绳胺酸、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、半胱胺酸、酿胺酸、 20天冬賴、麵胺、天冬酸、麵胺酸、離胺酸、精胺酸、及組胺酸。於較佳實施例中，胺基酸為L-組態。另外，胺基酸可為丙胺酿基、線胺醯基、白胺醯基、異白胺醯基、脯胺醯基、苯基丙胺醯基、色胺酸基、蛋胺醯基、甘胺酿基、 4胺&&基、蘇胺醯基、半胱胺醯基、路胺酿基、天冬酿胺 29 200831525 基、麵胺基、天冬醯基、戊二醯基、離胺醯基、精胺醯基' 組胺醯基、万_丙胺醯基、纈胺醯基、白胺醯基、召· 異白胺醯基、万-脯胺醯基、点—苯基丙胺醯基、万，色胺醯基、石-蛋胺醯基、召-甘胺醯基、点―絲胺醯基、冷―蘇胺醯 · 5基、召·半胱胺醯基、召路胺醯基、/5 _天冬醯胺基、/5-麵胺基、天冬醯基、点_戊二醯基、々_離胺醯基、万―精胺、醯基、或/3-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 '· 括但非限於呈D組態及L組態之α、冷、τ或5甘胺酸、丙 _ 1〇胺酸、纈胺酸、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、半胱胺酸、酪胺酸、天冬胺、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 - 括加成至氧、氮、硫或碟原子來防止其進一步反應或用於 · 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等人，有機合成保護鲁基，約翰威利父子公司）。除非另行陳明，芳基一詞如此處使用包括苯基、聯苯基或萘基且較佳為笨基。芳基視需要可經以一個或多個部取代忒等取代部分諸如鹵原子、魏基、胺基、烧基胺 ^、芳基胺基、烷氧基、芳氧基、硝基、氮基、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，例如如參考Greene等人， 30 200831525 有機合成保護基，約翰威利父子公司教示。烷芳基或烷基芳基等詞包括有芳基取代基之烷基。芳炫基或务基燒基專詞包括有烧基取代基之芳基。鹵原子一詞用於此處包括氯、溴、碘、及氟。 5 醯基一詞包括羧酸酯，其中酯基之非羰基部分係選自於直鏈、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、芳燒基包括节基、芳氧基烷基諸如苯氧基甲基、芳基包括視需要可經以鹵素、CiSC4烷基或(^至匕烷氧基取代之苯基、磺酸酯諸如烷基磺醯基或芳烷基磺醯基 10包括曱磺醯基、一磷酸酯、二磷酸酯、或三磷酸酯、三苯甲基或一甲氧基三苯甲基、經取代之节基、三烷基矽烷基 (例如二甲基·第三丁基矽烷基）或二苯基甲基矽烷基。酯之芳基最佳包含苯基。「低碳醯基」一詞係指其中非羰基部分為低碳烧基之酸基。 15 如此處使用，就對映異構純度而言，「實質上不含」或「貫質上不存在有」一詞係指一種組成物包括至少85%或 90/。重里比，較佳95%至98%重量比，又更佳99%至100%重里比所指疋之對映異構物。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。 20 同理’「分離」一詞係指一種化合物組成物包括至少 85%或90%重量比，車交佳95%至98%重量比，及又更佳㈣至100%重ϊ比之化合物，差額包含其它化學種類或對映異構物。「各自分別」一詞用於此處係指分開施用之變數，該 31 200831525 5 10 變數因應用用途而異有獨立變化。之化合物中，其中八 fc，於諸如R，，XYIT兩個R，，可為氫，戈γ刀別為石灭我氦J，兩個R”可為碳，了為見，或-她，，為碳而巧火「藥學切射藥」用來說明化合物之㈣上可接受II於-文說明書中上;接其當投予病人時可_化合物。藥學及盈機酸戍生自藥學上可接受之無機驗或有機驗 ‘…或有機酸之鹽。適當鹽類包括衍生自驗金屬及納之鹽及衍生自驗土金屬如約及鎂之鹽，多種其它酸為熟者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、，Figure 9 shows that the cisplatin (CDDP) resistant cell line endogenously exhibits superactive Akt. Figure 9A shows a line graph of two ovarian cancer cell lines (A2780S 23 200831525 and OV2008) and their western platinum-resistant dual (respectively A2780CP and C13) treated with increasing amounts of cepacidine. The Westbinder resistance mutant, A2780CP and C13, showed an increase in survival rate in response to West Platinum. The 9th figure shows that the phenotype of the ovarian cancer cells, ie, A2780S and OV2008, is compared with the resistance of the statin. The top panel shows superactive Akt in the West Platinum resistant cell line and A2780S. The figure below shows the total amount of Akt present in various types of cells. Figure 10 shows that Qucilide (TCN) (ΑΠ-2) overcomes the resistance of the West. The western platinum-resistant cell lines, namely A2780S and C13 lines, were treated with 〇, 5, 1 〇 and xin 10 20 μ Μ xiplatin. White striped bars indicate treatment with only platinum. The black ash column indicates treatment with dextroplatin and 10 μM. Co-treatment with TCN reduced cell viability and indicated that TCN overcomes the resistance of cetyltin. Figure 11 shows the synergistic effect of cell survival on C丨3 ovarian cancer cells using T C N and cetamine. C13 cells are treated with ... Bu Bu 1〇 & 2 (^mtcn - 15 containing and not containing ΐ〇μΜ West age. Xicai and Τ (: Ν composition can synergistically reduce cell viability. , I2 The figure shows that TCN can enhance the ability of Xicai to inhibit the growth of C13-implanted cancer cells in nude mice xenografts. Nude mice receive (3) xenografts, followed by vehicle (DMSO), sedative alone, alone TCN or Western Platinum (IV) TCN treatment 'Subsequent analysis of tumor regression after 7, 4, 21, or hour after inoculation. Left panel (A) shows a one-line diagram to verify the significant (four) tumor growth of the combination of Xi Mingting and tcn. Right _) display Tumor quality in vaccinated mice receiving representative treatment. Figure 1, the rapid activation of his path-inhibiting inhibitor 24 200831525. The above figure (A) shows the use of 1 nM mTOR West dot analysis of lysate of OV3 cells after treatment with the inhibitor of rapamycin. Membrane is probed for phosphorylated isoforms and antibodies specific for total protein for Akt, p70S6K Comparison with FKHRL 1. Lower - 5 Figure (B) shows the use of mTOR inhibitors Western blot analysis of MCF7 cell lysate after HAD001 treatment for a specified period of time. Membrane is used for phosphorylation...Isotypic and antibody specific for total protein for Akt and p70S6K comparisons. • Figure 14 shows The activation of the AKT pathway through the mTOR inhibitor was attenuated by 10 TCN. The left panel shows the effect of TCN on Akt phosphorylation treated with rapamycin in 〇V3 cells. The right panel (B) shows the TCN pair in MCF7 cells. The same effect of Akt phosphorylation treated with RAD001. Figure 15 shows the effect of cell growth on the combination of TCN with RAD001 or rapamycin. The top left panel (A) is a line graph showing the control group, * 15 alone The number of cells in DU-145 cells treated with rapamycin, TCN alone or TCN and rapamycin. The top right panel (B) is a line graph showing the control group, RAD001 alone, TCN alone or TCN and RAD001. The number of cells in MCF7 cells was treated on the 6th day. The following figure (〇 is a line graph showing the number of cells in the OV3 20 cells treated with the control group, laparomycin alone, TCN alone or TCN and rapamycin for 6 days. Figure 16 shows via Aurora (Aurora -A activates Akt to induce cell survival and chemoresistance. The above figure (A) shows that Aurora-A successfully transferred infection and expression in A2780S, A2780CP and OV2008 ovarian cancer cell lines. Figure 2 (B) is a line graph It is shown in A2780S cells that Aurora-A enhances cell viability and resistance to paclitaxel. The third panel (C) is a line graph showing that Aurora-A enhances cell viability and resistance to cetillin in A2780S cells. The following figure (D) shows that Aurora-A enhances cell viability and resistance to cetidine in 〇¥2〇〇8 cells. · 5 Figure 17 shows the effect of Aurora-A on cytochrome c release and Akt activation. Figure 17A shows immunofluorescence of cytochrome c in A2780S cells infected with a blank vector or with Aurora-A metastasis treated with or without western platinum. The release of cytochrome c induced by uranyl is inhibited by Aurora-A. Figure 17B shows that the western blot shows that the performance of Aurora _A can be increased in the phosphorylation of Akt in the cells of West Platinum and Tingmin. Figure 17C shows that increasing the concentration of Aurora-a increases the phosphorylation and activation of Akt. Figure 18 shows the effect of TCN (API-2) on Aurora-8 induction of West-platin. The first two panels (A and B) show the infection with a blank vector or Aurora-A transfer, followed by the use of cetrolidine alone, API-2 or API-2 alone and cisplatin, and A2780S cells. The next two panels (C and D) show infection with a blank vector or Aurora *-A transfer, followed by cisplatin alone, API-2 or API-2 alone, and OV2〇〇8 cells treated with serotonin or butyl. TCN was reduced in Western Platinum and Ting resistance provided by Aurora-A performance in both 278〇§ and 〇乂2〇〇8 cells. Figure 19 shows a schematic representation of the role of API-2, Aurora-A, p53, and Akt in cell survival. Figure 20 shows the effect of the Src family tyrosine kinase inhibitor dasatinib (Sprycel) on § ^ ^ ^ and Akt phosphorylation in eight 375 cells. Inhibition of Src tyrosine kinase increases the acidification of Akt. 26 200831525 • 5 [Embodiment 3] Detailed Description of the Preferred Embodiments In contrast to the prior art and experience, the inventors have determined how to successfully use a tripecline compound in combination with one or more compounds in one of the following ways or a combination thereof. Tumors and Cancers: (1) Trifluralin and one or more platinum compounds are administered only to patients who are more sensitive to the tricinemide compound and/or one or more platinum compounds according to the diagnostic test described later; (ii) the use regulations a dose level that minimizes the toxicity of the tricinemide compound and/or one or more germinal compounds while still having efficacy; or (iii) uses the 10-drug regimen to minimize Quci Toxicity of the compound and/or one or more platinum compounds. 5.1 Definitions As used herein, the term "compounds of the invention" is used to encompass compounds of formula I-IV, formula V, structural formula A-quinone compounds, and combinations thereof. ^ 15 As used herein, the terms "cancer" and/or "cancerous" describe or describe a physiological condition characterized by the growth of unregulated cells in a mammal', that is, 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, 20 colon cancer, squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, gastrointestinal cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer such as liver cancer , bladder cancer, colorectal cancer, endometrial cancer, kidney cancer and thyroid cancer. Non-limiting examples of other cancers are basal cell carcinoma; cholangiocarcinoma; bone cancer, brain and middle nervous system (CNS) cancer; choroidal carcinoma; connective tissue cancer; 27 200831525 esophageal cancer: eye cancer; head and neck cancer; Intraepithelial neoplasms; pharyngeal carcinoma; lymphoma including Hodgkin's lymphoma and non-Hodgkin's lymphoma; melanoma; myeloma; neuroblastoma; oral cancer (eg lip, sputum, π B preparation _ s, ± square, mouth and throat), pancreatic cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; respiratory cancer; 5 sarcoma; skin cancer; gastric cancer; testicular cancer, uterine cancer; urinary system cancer and other cancer Tumor and sarcoma. As used herein, the term "tumor" refers to all neoplastic cell growth and proliferation, including malignant or benign and all precancerous and cancerous cells and tissues. For example, a particular cancer can be characterized by a solid mass of tumor. A solid tumor mass can be a primary tumor mass when present. Primary tumor mass refers to the growth of cancer cells within the tissue resulting from normal cell transformation of the tissue. In most cases, primary tumor masses are identified by the presence of cysts, which can be found by visual methods or palpation methods, or by the irregular shape, texture, or weight of the tissue. However, several primary tumors cannot be palpated and can only be detected by medical imaging techniques such as X-rays (such as breast photography) or by needle aspiration. The use of the techniques described later is more common in early detection. Molecular analysis and phenotypic analysis of cancer cells within the tissue can often be used to verify whether the cancer is cancer within the tissue, or whether the lesion is from another site of cancer metastasis. 20 As used herein, unless otherwise indicated, the term alkyl includes, for example, straight, branched or cyclic first hydrocarbons, second hydrocarbons, or third hydrocarbons, such as Cl to C24, specifically including methyl, trifluoromethyl. , ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, pentyl, cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl, Cyclohexylmethyl, 3-methylpentyl, 200831525 2,2-dimethylbutyl, and 2,3-dimethylbutyl. The alkyl group may be optionally substituted, for example, with one or more substituents such as a halogen atom (ρ, ci, Br or I) (for example, cf3, 2-ethylidene, ch2F, CH2a, CH2CF3, or CF2CF3), Per group (eg CH2〇H), amine group (eg CH2NH2, 5ch2nhch3' or CH2N(CH3)2), alkylamino, arylamine, alkoxy, aryloxy, nitro, azide (eg CH2N3), cyano (eg WCH2CN), mineral acid, sulphate, phosphonic acid, phosphate, or phosphonate, either unprotected, or as known to those skilled in the art, as needed, for example, Greene Et al., Organic Synthetic Protection, 1991, 2nd ed., John Wiley & Sons, 10 New York teach, incorporated herein by reference. As used herein, the term lower alkyl, as used herein, refers to a straight-chain, branched, or, if appropriate, cyclic (eg, cyclopropyl) alkyl group, including substituted forms and Unsubstituted form. The term "alkylamino" or arylamino includes the radicals having one or two alkyl or aryl substituents. The term amino acid includes both natural and synthetic alpha, /5, r. Or a base acid, including but not limited to amino acids present in proteins, namely glycine, alanine, lysine, leucine, isoleucine, methionine, phenylalanine, tryptamine Acid, valine, serine, threonine, cysteine, tyrosine, 20 days of winter, face amine, aspartic acid, face acid, lysine, arginine, and histamine In a preferred embodiment, the amino acid is in the L-configuration. In addition, the amino acid can be a propylamine aryl group, a linear amine sulfhydryl group, an arginyl fluorenyl group, an iso-amine amide group, an amidoxime group, Phenylalanine oxime, tryptophanyl, egg amine sulfhydryl, glycine broth, 4 amine && base, sulphate, cysteamine, sulphate, aspartame 29 200831525 , face amine, aspartame, pentane, guanidinium, arginyl thiol 'histamine thiol, phenanthrenyl amide, amidoxime, leucine, singular Amidoxime, phenanthrenyl fluorenyl, phenyl-phenylamine hydrazino, phenanthrene, tryptamine sulfhydryl, sulphate-glycosyl fluorenyl, s-glycine thiol, s-silamine sulfhydryl, cold-sulphide Amine 5·5 base, cyanocysteine, hydrazine, /5 _ aspartate, /5- face amine, aspartate, _ 戊醯, 々 Derivatives of amidoxime, virginamine, sulfhydryl, or /3-histamine thiol. When the term amino acid is used, it is regarded as the natural amino acid or the synthetic amino acid ester. Specific independent disclosure, package '· including but not limited to alpha, cold, tau or 5 glycine, propionate, prolyl, leucine, isoleucine in D configuration and L configuration , methionine, phenylalanine, tryptophan, valine, serine, threonine, cysteine, tyrosine, aspartame, face amine, aspartic acid, glutamic acid, Amino acid, arginine, and histidine. Unless otherwise defined, "protected as used herein." The term package includes groups that are added to oxygen, nitrogen, sulfur or dish atoms to prevent further reactions or for other purposes. A wide variety of oxygen and nitrogen protecting groups are organic synthesis techniques, and those skilled in the art have Know (see Greene et al., Organic Synthesis Protection, Luki, John Wiley & Sons). Unless otherwise stated, the term aryl is used herein to include phenyl, biphenyl or naphthyl and is preferably stupid. The base is required to be substituted with one or more moieties such as a halogen atom, a thiol group, an amine group, an alkylamine group, an arylamine group, an alkoxy group, an aryloxy group, a nitro group, a nitrogen group, or the like. Sulfonic acid, sulfate, phosphonic acid, phosphate or phosphonate, either unprotected or as known to those skilled in the art, may be protected as needed, for example, as described in Greene et al., 30 200831525 Organic Synthetic Protecting Group, John Willie Father and son company teaching. The terms alkaryl or alkylaryl include alkyl groups having an aryl substituent. The aryl or ketone group includes an aryl group having a substituent. The term halogen atom is used herein to include chlorine, bromine, iodine, and fluorine. The term "fluorenyl" includes carboxylic acid esters wherein the non-carbonyl moiety of the ester group is selected from linear, branched or cyclic alkyl or lower alkyl, alkoxyalkyl including methoxymethyl, aryl The alkyl group includes a benzyl group, an aryloxyalkyl group such as a phenoxymethyl group, and an aryl group including a phenyl group, a sulfonate such as an alkyl group which may be optionally substituted by halogen, CiSC4 alkyl or (^ to alkoxyl group). Sulfoinyl or aralkylsulfonyl 10 includes sulfonyl sulfhydryl, monophosphate, diphosphate, or triphosphate, trityl or monomethoxytrityl, substituted benzyl, a trialkylalkylene group (for example, dimethylbutoxybutylenealkyl) or a diphenylmethyldecylalkyl group. The aryl group of the ester preferably contains a phenyl group. The term "low carbon fluorenyl group" means a non-carbonyl group thereof. Part of the acid group of a low carbon alkyl group. 15 As used herein, the term "substantially free" or "permanently absent" in terms of enantiomeric purity means that a composition includes at least 85%. Or 90/. by weight ratio, preferably 95% to 98% by weight, and more preferably 99% to 100% by weight of the enantiomer of the indicated. In a preferred embodiment, in the present invention In the methods and compounds, the compound is substantially free of other enantiomers. 20 Similarly, the term "separation" means that a compound composition includes at least 85% or 90% by weight, and the car is preferably 95% to 98%. Weight ratio, and more preferably (d) to 100% by weight of the compound, the difference includes other chemical species or enantiomers. The term "respectively separate" is used herein to mean a variable to be applied separately, the 31 200831525 5 10 Variables vary independently depending on the application. Among the compounds, eight fc, such as R, XYIT, two R, can be hydrogen, Ge γ knife is stone, I, J, two R" For carbon, for the sake of seeing, or - her, for the carbon and fire "pharmaceutical injection" to indicate the compound (4) acceptable II in the manual; when it is administered to the patient can be _ compound Pharmacy and sputum sputum is derived from a pharmaceutically acceptable inorganic or organic test.... or a salt of an organic acid. Suitable salts include salts derived from metals and salts and derived from soils such as magnesium and magnesium. Salt, a variety of other acids known to the pharmaceutical industry, is well known. Pharmaceutically acceptable A 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 a functional moiety of the active compound. Redox, amination, deamination, hydroxylation, dehydroxylation, hydrolysis, dehydrolysis,

15 化 >丁、取ΓΊυ、云胺化、羥化、去羥亿、不解、紊水解、$ 、去烧化、酿化、去醯化、磷酸化、去填酸化來製造性化合物之化合物「藥學上可接受之酯類」〆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内，適合用來與宿主組織接觸而無不當毒 20 性、刺激性、過敏反應等，具有合理效益/風險比，可有效用於其期望用途。「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、驟、馬、猶及其它非人靈長類、15] butyl, oxime, cloud amination, hydroxylation, dehydroxylation, insoluble, turbid hydrolysis, $, de-sintering, brewing, deuteration, phosphorylation, de-acidification to produce compounds The term "pharmaceutically acceptable ester" is used herein to include, unless otherwise stated, esters of one or more compounds which are suitable for use in contact with host tissues within the context of prudent medical judgment. No irrational toxicity, irritant, allergic reaction, etc., with reasonable benefit/risk ratio, can be effectively used 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, spurs, horses, and other non-human primates,

32 200831525 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及相關化合物組合一種或多 5 種顧化合物用於治療增生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：32 200831525 A canine, cat, rat, mouse, rabbit or any other mammal known or as disclosed herein. 5.2 Compounds of the Invention The present invention provides the use of TCN, TCN_P and related compounds in combination with one or more of the compounds of the formula for the treatment of 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:

10 其中R2’、R3’及R5’分別為氫、視需要可經取代之磷酸 33 200831525 根或鱗酸根（包括一鱗酸、二鱗酸、或三填酸或安定化鱗酸前藥）；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 5 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<提供一種化合物其中R2,、R3,或R5,分別為Η或一磷酸、；璘酸、或三磷酸； 10 其中Rx及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、去氧伸烧基諸如聚乙二醇、視需要可經取代之芳基讀醯基；脂質包括磷脂質；胺基酸；碳水化合物；胜肽；戒膽固醇；或其它藥學上可接受之離去基；於一個實施例中’ 15 該化合物係呈5’-磷醚脂質或5，-醚脂質投藥。10 wherein R2', R3' and R5' are respectively hydrogen, optionally substituted phosphoric acid 33 200831525 root or sulphate (including mono sulphate, di squaric acid, or tri-acid or stabilized sulphate prodrug); Sulfhydryl (including lower fluorenyl); alkyl (including lower alkyl); decylamine, sulfonate including alkyl or arylalkyl; sulfonyl including methanesulfonyl and benzyl, wherein the benzene The base view may be substituted with one or more substituents as described in the definition of the aryl 5 group described herein; optionally substituted arylsulfonyl; lipids including phospholipids; amino acids; carbon water a compound; a peptide; or a cholesterol; or other pharmaceutically acceptable leaving group, in vivo <providing a compound wherein R2, R3, or R5, respectively, hydrazine or monophosphate; citric acid, or Phosphoric acid; 10 wherein Rx and Ry are respectively hydrogen, optionally substituted phosphate; sulfhydryl (including low carbon fluorenyl); decylamine, alkyl (including lower alkyl); aromatic, deoxygenated Base such as polyethylene glycol, optionally substituted aryl read thiol; lipids including phospholipids; amino acids Carbohydrate; peptide; cholesterol ring; or on other pharmaceutically acceptable leaving group; Example '15 This compound was 5'-phosphorus-based ether lipid to one embodiment 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、或環狀烷基（包括低碳烷基）、烯基或快基、CO-烷基、烯基、CO-炔基、CO-芳基或雜芳基、co_烷氧基烷基、C〇芳氧基烧基、CO-經取代之芳基、磺醯基、烷基磺醯基、芳 20 基磺醯基、芳烷基磺醯基。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,、m，為氮。於又另一個實施例中，R2’、r3,、r5,、Ri&r2為氫。於另一個實施例中，曲西立濱化合物具有如下結構式： 34 200831525Each of Ri and R2 is independently a linear, branched, or cyclic alkyl group (including a lower alkyl group), an alkenyl group or a fast group, a CO-alkyl group, an alkenyl group, or a CO-alkynyl group which may be substituted. , CO-aryl or heteroaryl, co-alkoxyalkyl, C aryloxyalkyl, CO-substituted aryl, sulfonyl, alkylsulfonyl, aryl 20 sulfonyl , aralkyl sulfonyl group. In one embodiment, R2' and R3 are hydrogen. In another embodiment, r2' and r5' are hydrogen. In yet another embodiment, R2, m is nitrogen. In yet another embodiment, R2', r3, r5, Ri&r2 are hydrogen. In another embodiment, the triclinide compound has the formula: 34 200831525

其中I為H、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、芳基、烧氧基烷基、芳氧基烷基、或經取代之芳基；以及 5 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、烧乳基、烧氧基烧基、毯基烧基、或方基。於一次實施例中，R3為直鏈ci-ii烷基、異丙基、第三丁基或苯基。 10 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 I is H, optionally substituted straight chain, branched or cyclic alkyl (including lower alkyl), alkenyl, or alkynyl, NH2, NHR4, N(R4)2, aryl, oxygenated An alkyl group, an aryloxyalkyl group, or a substituted aryl group; and 5 each R 4 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, a calcined base group, an alkoxy group, a carpet base group, or a aryl group. In one embodiment, R3 is a linear ci-ii alkyl, isopropyl, tert-butyl or phenyl group. In one embodiment, the triclinide compound provided herein has the formula:

35 200831525 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：35 200831525 In another embodiment, the triclinide compound provided herein has the following structural formula:

5 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：In another embodiment, the triclinide compound provided herein has the following structural formula:

其中R6為Η、炫基（包括低碳烧基）、稀基、快基、烧氧基烷基、羥基烷基、芳基烷基、環烷基、ΝΗ2、NHR4、NR4R4、 CF3、CH2OH、CH2F、CH2C1、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，此處各個Y3分別為H或鹵原子；以及 36 10 200831525 各個R4分別為Η、醯基包括低碳酸基、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、環烷基、烷氧基、烷氧基烷基、羥基烷基、或芳基。於一次實施例中’ 116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 anthracene, stilbene (including low carbon alkyl), dilute, fast radical, alkoxyalkyl, hydroxyalkyl, arylalkyl, cycloalkyl, hydrazine 2, NHR4, NR4R4, CF3, CH2OH, CH2F, CH2C1, 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, where each Y3 is H or halogen Atom; and 36 10 200831525 Each R4 is fluorene, fluorenyl, including lower alkyl, decyl including low carbon alkyl such as, but not limited to, methyl, ethyl, propyl, and cyclopropyl, alkenyl, alkynyl, A cycloalkyl, alkoxy, alkoxyalkyl, hydroxyalkyl, or aryl group. In one embodiment '116 is ethyl, CH2CH2OH or CH2-phenyl. In another embodiment, the triclinide compound provided herein has the following structural formula:

其中R7為Η、函原子、烷基（包括低碳烷基）、烯基、炔基、烷氧基、烷氧基烷基、羥基烷基、環烷基、硝基、氰基、OH、OR4、厕2、NHR4、NR4R4、SH、SR4、CF3、CH2〇H、 ch2f、ch2ci、ch2cf3、c(y3)3、c(y3)2c(y3)3、C(=〇)〇H、 C(二0)0R4、C(=0)-烧基、c卜0)-芳基、C(=0)-院氧基烷基、 C(=0)NH2、C(=〇)NHR4、C(=0)N(R4)AN3，此處各個 γ3 分別為11或_原子；以及各個R分別為Η、醯基包括低碳醯基、烧基包括低碳院基諸如但非限於曱基、乙基、丙基、及環丙基、烯基、炔基、環烧基、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。 37 200831525 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 functional atom, an alkyl group (including a lower alkyl group), an alkenyl group, an alkynyl group, an alkoxy group, an alkoxyalkyl group, a hydroxyalkyl group, a cycloalkyl group, a nitro group, a cyano group, an OH group, OR4, toilet 2, NHR4, NR4R4, SH, SR4, CF3, CH2〇H, ch2f, ch2ci, ch2cf3, c(y3)3, c(y3)2c(y3)3, C(=〇)〇H, C (2) 0R4, C(=0)-alkyl, cBu0)-aryl, C(=0)-homoyloxyalkyl, C(=0)NH2, C(=〇)NHR4, C (=0) N(R4)AN3, where each γ3 is 11 or _ atoms, respectively; and each R is Η, 醯, including a low carbon fluorenyl group, and the alkyl group includes a low carbon yard group such as, but not limited to, a fluorenyl group, Ethyl, propyl, and cyclopropyl, alkenyl, alkynyl, cycloalkyl, alkoxy, alkoxyalkyl, hydroxyalkyl, or aryl. In one embodiment, R7 is methyl, ethyl, phenyl, chloro or hydrazine. 37 200831525 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:

Cl%Cl%

如此處使用且除非另行指示，「鉑化合物」一詞係指式 38 200831525 v化合物： I白化合物As used herein and unless otherwise indicated, the term "platinum compound" refers to the formula 38 200831525 v Compound: I White Compound

Hi % %Hi % %

式V 二 5其中： R!、R2、R3及R4各自分別為氫、視需要可經取代之胺、 Φ 芳香族胺、雜芳香族胺、視需要可經取代之直鏈、分支或 ί哀狀烧基、Ri、R2、R3及R4間所形成之芳香環、雜芳香環或環狀環。Formula V 2: wherein R!, R2, R3 and R4 are each independently hydrogen, optionally substituted amine, Φ aromatic amine, heteroaromatic amine, linear chain, branch or sorrow which may be substituted as needed An aromatic ring, a heteroaromatic ring or a cyclic ring formed between Ri, R2, R3 and R4.

10 於該具體實施例中，鉑化合物包括但非限於結構式A-I 之鉑化合物：In this particular embodiment, the platinum compound includes, but is not limited to, a platinum compound of the formula A-I:

NH3 NHg mNH3 NHg m

39 200831525 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)組態或可為其混合物。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疑光形式’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： (i) 晶體之物理分離-可手動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； (111)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-至少一個合成步驟係使用 0崎催化反應來獲得期望之對映異構物之對映異構純質或對映異構豐富合成前驅物之合成技術； (V)化本非對稱性合成-於可於產物中產生非對稱性 (亦即對掌性)之條件下，由非對倾驅物合成射之對映異冓物之泛成技術’该條件可使用對掌催化劑或對掌輔劑來 40 200831525 達成； …（卜對映異構物分離·外、消旋化合物與對映異構純。κ f革輔奶反應’將個別對映異構物轉成非對映異構物 ;ϊ所得非對映異構物隨後由於現在已經變成有較明 5顯的結構差異，故藉層析術或藉結晶分離，對掌輔劑隨後被去除來獲得期望之對映異構物； (νΐ1)第-級及第二級非對稱性轉換_來自於外消旋物之非對映異構物平衡來於得自期望之對映異構物之非對映異構物办體中獲得優勢，或得自期望對映異構物之非對映 10異構物優先結晶擾亂平衡，最終原則上全部材料皆轉成得自期望之對映異構物之結晶性非對映異構物之技術。期望之對映異構物隨後由非對映異構物中釋放； (观）動態光學分割_本技術係指於動態條件下，藉由對映異構物與對掌非外消旋試劑或催化劑之不等反應速率， b來達成外消旋物之部分光學分割或完全光學分割（或部分光學分割化合物之進一步光學分割）； (ix)由非外消旋前驅物之對映異構特1性人成-由非對掌起始物料獲得期望之對映異構物之_種合成技術，此處立體化學之完好不因合成過程受損，或只有極少受損； 2〇 (X)財液相層析術-外消旋物之對映異構物藉由其與靜相間之差異交互作用而被分離入液體動相之技術。靜相係由對掌材料所形成，或動相可含有額外對掌材料來激發差異交互作用；㈣對掌氣相層析術-外消旋物氣化，對映異構物藉由 41 200831525 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_藉由一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¥膜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，一種含有外消旋物，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: p. 4757-4760。 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：Gudmiindsson 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 42 20083152539 200831525 It is to be understood that the compounds disclosed herein may contain a palm center. Such a palm center can be either (R) configured or (S) configured or can be a mixture 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 racemic form, optically active form, polymorphic form or stereoisomeric form, or mixtures thereof, preferably having the useful properties described herein, and the art is well known in the art of how to prepare a suspected form. How to use the standard assays described herein to determine activity, or to use other similar assays known to the art. Examples of methods which can be used to obtain optical isomers of the compound include the following: (i) Physical separation of crystals - a technique for manually 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 the individual enantiomers separately from the racemic solution by crystallization; (111) Enzymatic catalyzed optical cleavage _ via the different reaction rates of the enantiomer and the enzyme to achieve elimination a technique for partial or complete separation of a spirulina; (iv) enzymatic catalyzed asymmetric synthesis - at least one synthetic step using a zero-synthesis reaction to obtain the enantiomerically pure or enantiomer of the desired enantiomer Synthetic technology of heterogeneous and rich synthetic precursors; (V) asymmetry synthesis - under the condition that asymmetry (ie, palmarity) can be produced in the product, the pair is synthesized by non-dip The generalization technique of the sputum sputum 'this condition can be achieved using the palm catalyst or the palm adjuvant 40 200831525; ... (the enantiomer separation, the outer, the racemic compound and the enantiomerically pure. κ f Leather-assisted milk reaction 'converts individual enantiomers to diastereomers The resulting diastereomers are subsequently converted to a clearer 5 structural difference, whereby the palm adjuvant is subsequently removed by chromatography or by crystallization to obtain the desired enantiomer; (νΐ1) Stage- and second-stage asymmetry conversions - from the diastereoisomer balance of the racemates in the diastereomers from the desired enantiomer The advantage is obtained, or the diastereomeric 10 isomers from the desired enantiomers are preferentially crystallized to disturb the equilibrium. Finally, in principle all materials are converted into crystalline diastereoisomers derived from the desired enantiomer. Technology of the object. The desired enantiomer is subsequently released from the diastereomer; (view) dynamic optical segmentation - this technique refers to the enantiomeric and non-external The unequal reaction rate of the racemic reagent or catalyst, b to achieve partial optical splitting or complete optical splitting of the racemate (or further optical splitting of the partially optically split compound); (ix) by a pair of non-racemic precursors Amino-specific, from the non-palm starting material to obtain the desired pair The synthesis technique of isomers, where the stereochemistry is intact and not damaged by the synthesis process, or only minimally damaged; 2〇(X)Qi liquid chromatography-enantiomer of racemate a technique of separating into a liquid mobile phase by interaction with a static phase. The static phase system is formed by the palm material, or the moving phase may contain additional palm material to excite the differential interaction; Chromatography-racemate gasification, the enantiomer is separated by 41 200831525 in the gas phase with the difference between the column containing the fixed non-racemic to palm adsorption phase; Palm solvent extraction - a technique for separating enantiomers by dissolving one enantiomer in a specific solvent; 5 (Xm) technology for transporting racemates to membrane barriers across the membrane . The barrier typically separates two miscible fluids, one containing a racemate, 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, Tricinelbine, Tricineribine Phosphate (TCN-P), 5'-Trimethoprimate (TCN_p), or Trifluralin (TCN-DMF) is provided. The TCN can be synthesized by any of the techniques known to those skilled in the art, for example, in a tetrahedral letter, 1971.49: p. 4757-4760. 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: Gudmiindsson 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 42 200831525

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.4l_44;及/或Wotring LL·等人，癌症治療Rep, 1986. 70(4): 5 ρ·491·7 〇 5.3藥學上可接受之鹽、水合物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Townsend LB et al., Nucleic Acid Symp Ser, 1986·1986(17): p.4l_44; and/or Wotring LL. et al., Cancer Therapy Rep, 1986. 70(4): 5 ρ·491·7 〇5.3 Pharmacy The acceptable salts, hydrates and prodrugs are sufficiently basic or acidic to form stable non-toxic acid salts or bases.

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酸。適當鹽類包括衍生自鹼金屬諸如鉀及鈉、鹼土金屬諸如舞及鎮之鹽，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，例如甲苯磺酸鹽、甲磺酸鹽、乙酸鹽、檸檬酸鹽、丙二酸鹽、酒石酸鹽、丁二酸鹽、苯 15甲酸鹽、抗壞血酸鹽、酮基戊二酸鹽、及α-甘油基磷酸鹽。也可形成適當之無機酸鹽包括硫酸鹽、硝酸鹽、重碳酸鹽、及碳酸鹽。於另-個實施例中，曲西立濱化合物為溶劑合物例如磷酸曲西立濱一水合物。 20 _學上可接受之鹽也可使用技藝界眾所周知之標準程序獲得，例如充分驗性化合物諸如胺與適當之形成I理上可接受之陰離子之酸反應。也可製造_之驗金屬（例如鈉、鉀或鐘）鹽或驗土金屬（例如甸）鹽。此處所述之任-種核㈣可呈核誓酸前藥投予來提高 43 200831525 活性、生物利用率、安定性或以其它方式變更核苔之性質。多種核苷酸前藥配體為已知。大致上核苷之一麟酸、二填酸或三磷酸之烷化、醯化或其它親脂改性將可提高核苔酸之安定性。可置換填酸部分上之一個或多個氫之取代基之 - 5實例為烧基、芳基、類固醇、碳水化合物 '包括糖類、1，2_ ，二醯基甘油及醇類。多種取代基說明於R. J0nes&N. 〜In the case of a salt, a pharmaceutically acceptable salt is administered as a suitable compound. Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic bases or 10 acids. Suitable salts include salts derived from alkali metals such as potassium and sodium, alkaline earth metals such as maiden and towns, 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 tosylate, methanesulfonate, acetate, citrate, C. Diacid salts, tartrates, succinates, benzene 15 formates, ascorbates, ketoglutarate, and alpha-glyceryl phosphates. Suitable inorganic acid salts can also be formed including sulfates, nitrates, bicarbonates, and carbonates. In another embodiment, the triclinbine compound is a solvate such as trifluralin phosphate monohydrate. 20 _Studyly acceptable salts can also be obtained using standard procedures well known to the artisan, for example, by fully testing a compound such as an amine with an appropriate acid which forms an acceptable anion. It is also possible to produce a metal (for example sodium, potassium or bell) salt or a soil test metal (for example). Any of the cores (4) described herein may be administered as a nuclear antacid prodrug to increase the activity, bioavailability, stability, or otherwise alter the properties of the nucleus. A variety of nucleotide prodrug ligands are known. Substantially alkylation, deuteration or other lipophilic modification of one of the nucleosides of the nucleoside, the second acid or the triphosphate will increase the stability of the nucleate. Substituents for replacing one or more hydrogens on the acid-filling moiety - 5 examples are alkyl, aryl, steroid, carbohydrate 'including saccharides, 1,2_, dimercaptoglycerol and alcohols. A variety of substituents are described in R. J0nes & N. ~

Bischofberger，抗病毒研究 ’ 1995.27:p-l-i7。全部皆可與、所揭示之核苔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，_ · 10 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，較佳掺混於核苷製劑或親脂製劑之5’-0H位置之美國專利案之非限制性實例包括美國專利案5，149,794 (Yatvin 等人）；5，194,654 (Hostetler等人）；5,223,263 (Hostetler等人）；5,256,641 (Yatvin等人）；5,411，947 (Hostetler等人）； 15 5,463,092 (Hostetler 等人）；5,543,389 (Yatvin 等人）； 5,543,390 (Yatvin 等人）；5,543,391 (Yatvin 等人）；及 5,554,728 (Basava等人），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取代基或親脂製劑之外國專利申請案包括W0 89/02733、 20 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 91/19721。曲西立濱或相關化合物之衍生物之額外非限制性實例為含有如下公告案所述之取代基。此等經衍生之曲西立濱 44 200831525 - 5 • 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: p.2816-2820; Holy Α·於抗病毒藥物設計進階，Vol.I，De Clercq (編輯），JAI出版社，ρρ· 179-231; Hong C.L 等人，Biochem Biophys Rs Commun, 1979.88: p.1223-1229; Hong C.L等人，J Med Chem，1980. 28: p.171-177; Hostetler Κ·Υ·等人，J Biol Chem，1990· 266: p.11714-11717; Hostetler K.Y.等人，Antiviral Res，1994.24: p.59-67; Hostetler Κ·Υ·等人，抗微生物劑化學治療，1994· 38: 10 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 Chern Soc Perkin Trans，1984. I: p.1471-1474; Juodka Β·Α·及 Smart J·，Coll Czech Chem Comm，1974· 39: ρ·363-968; KataokaS·等人，核酸研究Sym ' 15 Ser，1989· 21: p.1-2; Kataoka S.等人，雜環，1991· 32: p.1351-1356; Kinchington D.等人，抗病毒Chem Chemother， 1992. 3: ρ·107-112; Kodama K.等人，日本癌症研究期刊， 1989， 80: p.679-685; Korty M.及 Engels J·， N auny n- S chmi e deb erg5 s Arch Pharmacol, 1979. 10: 20 ρ·103_111; Kumar A·等人，J Med Chem，1990· 33: p.2368-2375; LeBec C·及Huynh-dinhT·，四面體函件，1991· 32: ρ·6553_6556; Lichtenstein J·等人，J Biol Chem，I960. 235: p.457-465; Lucthy J·等人，Mitt Geg Lebensmittelunters Hyg，1981.72: p.131-133 (Chem· Abstr. 95，127093); 45 200831525Bischofberger, Antiviral Research ' 1995.27: p-l-i7. All can be combined with the disclosed nuclear moss to achieve the desired effect. In one embodiment, 'tuclide or a related compound is provided as a 5, 10-hydroxy hydroxy lipophilic prodrug. A suitable lipophilic substituent is disclosed which can be covalently incorporated into a nucleoside, non-limiting examples of which are preferably incorporated in the 5'-0H position of a nucleoside formulation or a lipophilic formulation, including U.S. Patent 5, 149,794 (Yatvin et al.); 5,194,654 (Hostetler et al.); 5,223,263 (Hostetler et al.); 5,256,641 (Yatvin et al.); 5,411,947 (Hostetler et al.); 15 5,463,092 (Hostetler et al.); 5,543,389 (Yatvin 5,543,390 (Yatvin et al.); 5,543,391 (Yatvin et al.); and 5,554,728 (Basava et al.), each of which is incorporated herein by reference. _ Revealing a lipophilic substituent or a lipophilic preparation that can be attached to the tromethine or related compound of the present invention. Patent applications include W0 89/02733, 20 W0 90/00555, W0 91/16920, W0 91/ 18914, WO 93/00910, W0 94/26273, WO/15132, EP 0 350 287, EP 93917054.4, and WO 91/19721. An additional non-limiting example of a derivative of triclinide or a related compound is a substituent as described in the following publication. Such derived triclinibs 44 200831525 - 5 • or related compounds can be used for the indications described herein, or as anti-viral agents, including as anti-HIV agents or anti-HBV agents. Ho DHW·, Cancer Res·, 1973 33: p.2816-2820; Holy 进·Advanced in Antiviral Drug Design, Vol.I, De Clercq (ed.), JAI Press, ρρ· 179-231; Hong CL Et al, Biochem Biophys Rs Commun, 1979.88: p. 1223-1229; Hong CL et al, J Med Chem, 1980. 28: p. 171-177; Hostetler Κ·Υ· et al, J Biol Chem, 1990·266 : p.11714-11717; Hostetler KY et al., Antiviral Res, 1994.24: p.59-67; Hostetler Κ·Υ· et al., Antimicrobial Chemotherapy, 1994· 38: 10 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 AS· et al, J Chern Soc Perkin Trans, 1984. I: p.1471-1474; Juodka Β·Α·and Smart J·, Coll Czech Chem Comm, 1974· 39: ρ·363-968; KataokaS· et al., Nucleic Acids Research Sym ' 15 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: ρ·107-112; Kodama K. Et al, Journal of Cancer Research in Japan, 1989, 80: p.679-685 Korty M. and Engels J., N auny n- S chmi e deb erg5 s Arch Pharmacol, 1979. 10: 20 ρ·103_111; Kumar A· et al, J Med Chem, 1990· 33: p.2368-2375 LeBec C· and Huynh-dinhT·, tetrahedral letter, 1991· 32: ρ·6553_6556; Lichtenstein J· et al, J Biol Chem, I960. 235: p.457-465; Lucthy J. et al., Mitt Geg Lebensmittelunters Hyg, 1981.72: p. 131-133 (Chem. Abstr. 95, 127093); 45 200831525

McGuigan C·等人，核酸研究，1989· 17: p.6065-6075; McGuigan C.等人，抗病毒 Chem Chemother，1990. 1: p.107-113; McGuigan C.等人，抗病毒Chem Chemother，1990· 1: p.355-360; McGuigan C·等人，抗病毒Chem Chemother， 5 1990. 1: ρ·25-33; McGuigan C.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·Η)1; McGuigan C·等人，J Med Chem，1993. 36: p.1048-1052 。 10 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; 15 Nargeot J·等人，Natl. Acad. Sci. U.S.A·，1983· 80: p.2395-2399; Nelson K.A.等人，J Am Chem Soc，1987· 109: ρ·4〇58·4064; Nerbonne J.M·等人，自然，1984· 301: p.74-76;Neumann J.M.等人，J Am Chem Soc，1 1989.111: p.4270-4277; Ohno R.等人，腫瘤學，1991. 48: p.451_455;McGuigan C. et al., Nucleic Acids Res., 1989, 17: p. 6065-6075; McGuigan C. et al., Antiviral Chem Chemother, 1990. 1: p. 107-113; McGuigan C. et al., Antiviral Chem Chemother , 1990· 1: p.355-360; McGuigan C· et al., Antiviral Chem Chemother, 5 1990. 1: ρ·25-33; McGuigan C. 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·Η)1; McGuigan C· et al. J Med Chem, 1993. 36: p.1048-1052. 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. Human, J Med Chem, 1992· 35: ρ·3939-3044; 15 Nargeot J. et al., Natl. Acad. Sci. USA·, 1983· 80: p. 2395-2399; Nelson KA et al., J Am Chem Soc, 1987· 109: ρ·4〇58·4064; Nerbonne JM· et al., Nature, 1984· 301: p. 74-76; Neumann JM et al., J Am Chem Soc, 1 1989. 111: p. 4270-4277 Ohno R. et al., Oncology, 1991. 48: p.451_455;

20 Palomino Ε·等人，J Med Chem，1989· 32: ρ·622-625; Perkins R.M·等人，抗病毒研究，1993· 20 (Suppl.I): ρ·84; Piantadosi C.等人，J Med Chem, 1991· 34: 1408-1414; Pompon A.等人，抗病毒Chem Chemother，1994· 5: ρ·91-98; Postemark T·， Anu Rev Pharmacol，1974· 14: p.23-33; Prisbe E.J.等人，J 20083152520 Palomino Ε· et al, J Med Chem, 1989· 32: ρ·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; Postemark T·, Anu Rev Pharmacol, 1974· 14: p.23-33 ; Prisbe EJ et al, J 200831525

- 5 • Med Chem，1986. 29: ρ·671·675; Pucch F.等人，抗病毒研究，1993. 22: ρ·155_174; Pugaeva VeR等人，Gig Trf Prof Zabol，1969，13: ρ·47-48 (Chem· Abstr. 72, 212); Robins R.K·，Pharm Res，1984· 11-18; Rosowsky A·等人，J Med Chem，1982· 25: p.171-178; Ross W.，Biochem Pharm，1961. 8: p.235-240; Ryu Ε·Κ·等人，J Med Chem，1982. 25: p.1322-1329; Saffhill R.及Hume WJ·，Chem Biol.Interact， 1986. 57: p.347-355; Saneyoshi M·等人，Chem Pharm Bull， 1980. 28: ρ·2915-2923; Sastry J.K.等人，Mol Pharmacol， 10 1992· 41: p.441-445; Shaw J.R 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，Chem Pharm Bull，1988. 36: p.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」。 15 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、 20 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、5,512,671、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 47 200831525 93/08807 。 5·4活體内功效/投藥計刻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 - 5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、南也糖、σ區吐、低血妈、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、高、腹渴、胃炎及/或發燒。於另一個實施例中，投予TCN、TCN-P或相關化合物籲 10及一種或多種鉑化合物提供於活體内於至少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、20、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15、15、20、25、30、35、40、 15 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。 Φ 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-P、 20 TCN-PM或相關化合物及一種或多種鉑化合物投予該個體’該投藥計劃包括投予曲西立濱化合物及/或一種或多種鉑化合物每週一次計三週，接著為_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 '或5次，或重複至腫瘤之退行明顯為止。 48 200831525 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/或一種或多種鉑化合物之兩週時間。於其它實施例中，此種42天週期可重複至少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、TCN-PM或相關化合- 5 • Med Chem, 1986. 29: ρ·671·675; Pucch F. et al., Antiviral Research, 1993. 22: ρ·155_174; Pugaeva VeR 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.171-178; Ross W., Biochem Pharm, 1961. 8: p. 235-240; Ryu Ε·Κ· 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: ρ·2915-2923; Sastry JK et al., Mol Pharmacol, 10 1992· 41: p.441-445; Shaw JR 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: p. 209-217. A preferred phosphoric acid prodrug group is S-mercapto-2-thioethyl, also known as "SATE". 15 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, 20 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, 5,512,671, 6,030,960, 2004/0259845, 6,670,34, 2004/0161398, 2002/082242, 5,512,671, 2002/0082242 and/or PCT Publications WO 90/11079, WO 96/39197, and / or WO 47 200831525 93/08807. 5.4 In vivo efficacy/dosing schedule 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, the five-dosing schedule reduces or eliminates toxic side effects including, but not limited to, hepatotoxicity, thrombocytopenia, nanose, sigma sputum, hypoxemia, anemia, low albumin Hypertension, myelosuppression, hypertriglyceridemia in the blood, amylase in the blood, high, thirst, gastritis and/or fever. In another embodiment, administration of TCN, TCN-P or related compounds and one or more platinum compounds are provided in vivo for at least partial or complete reaction of at least the individual. In a particular embodiment, the partial response can be at least 15, 20, 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 15, 15, 20, 25, 30, 35, 40, 15 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. Φ In other embodiments, providing an individual who has been diagnosed with cancer by administering an effective amount of TCN, TCN-P, 20 TCN-PM or related compound and one or more platinum compounds to the individual according to a dosage regimen The dosing schedule includes administration of the triclinide compound and/or one or more platinum compounds once a week for three weeks, followed by _ weeks of undosing (ie, through a 28 day period). In other embodiments, such a 28 day cycle can be repeated at least 2, 3, 4 ' or 5 times, or repeated until the tumor is significantly degraded. 48 200831525 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 two weeks of non-administration of the triclinide compound and/or one or more platinum compounds. . In other embodiments, such a 42 day cycle can be repeated at least 2, 3, 4, or 5 times, or repeated until the tumor is regressed significantly. In a particular embodiment, less than 12, less than 11 or less than 10 mg/m 2 of TCN, TCN-P, TCN-PM or related compounds

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、1〇、11毫克/平方米1〇^、丁(：沁1>、1(：^_刚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1¾克至約50毫克鉑化合物。於特定實施例中，了根據42天週期投予 1、5、1〇、15、2〇、25、30、35、40、 5〇、60、70、80、90、或1〇〇毫克鉑化合物。 15The substance can be administered according to a 42-day cycle. In other specific embodiments, 2, 3, 4, 5, 6, 7, 8, 9, 1 , 11 mg / m 2 〇 ^, D (: 沁 1 >, 1 (: ^ _ just or related The compound can be administered according to a 42 day period. In another specific embodiment, from about 13⁄4 grams to about 50 mg of the platinum compound is administered. In a particular embodiment, 1, 5, 1 , 15, etc. are administered according to a 42 day period. 2 〇, 25, 30, 35, 40, 5 〇, 60, 70, 80, 90, or 1 〇〇 gram of platinum compound.

仍力一调貫施例中，提供於個菔冶潦癌症之方法，經由對該铺投予-種給藥賴，每週—次投予1〇毫克 :米或以下之TCN、TCN-P或相關化合物及低於約則 -種或多齡化合物。於特定實施例中，每週一 0.5 1·5、2、2·5、3、3.5、4、4.5 5.5 20 7 ν 一，υ、6.5 •、、8.5、9、9.5或1G毫克/平方米如此處揭示之Τ( CN_P或相關化合物。於另一個特定實施例中，可每Γ:1、5，、15、20、25、3。、35、4。、5。^、、90、或100毫克鉑化合物。之化合物可於短時間單一大劑量於本發明之實施例中，此處揭示例如約〜卜…^加分鐘時間同時呈單二投予。於料實_巾，提供„㈣，I大劑量 -』/、甲礒等化合物 49 200831525 係透過速續輸注歷至少24、48、72、96或120小時時間同時投予。於芳干實施例中，透過連續注射或大劑量注射投予曲西立濱化合物及/或一種或多種銘化合物可於某個頻率重複，該投藥頻率至少··每週一次、每兩週一次、每三週 5 一次、每個月一次、每五週一次、每六週一次、每八週一次、每十通一次、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/或一種或多種鉑化合物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。投藥 10計劃可投予至少歷時：1、2、3、4、5、6、7、8、9、1〇、 11、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/或一種或多種翻化合物可根據此處揭示之任一種組合投予，例如曲西立濱化合物及/或一種或多種鉑化 15 合物可每週一次’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1〇曰。此種投藥接著為未投予曲西立濱化合物及/或一種或多種鉑化合物之相對應週期時間。 20 如此處揭示之TCN、TCN-P及相關化合物及一種或多種鉑化合物可以可有效造成腫瘤退行之數量投予病人。 TCN、TCN-P及相關化合物及一種或多種鉑化合物之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， 50 200831525 至少2、5、10、15、20、30或50毫克/平方米此處揭示之曲西立續化合物可技予一個體。於若干實施例中，至少約 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、1〇、π、15、17、20、25、30、35、 • 5 4〇、45、50、55、60、65、70、75、80、85、90、95、100、 150、165、175、200、250、300、或35〇毫克/平方米如此處揭示之TCN、TCN-P、TCN-PM或相關化合物可投予一個體。於若干實施例中，1〇、20、50、100、150、200、250、 φ 300、35〇或4〇〇毫克鉑化合物可投予一個體。 10 化合物之投予可根據此處揭示之任一種治療計劃進Still in a continuation of the application, it is provided in a method for cancer, by administering a dose of 1 〇 milligrams per meter or less to TCN, TCN-P. Or related compounds and less than about one or more compounds. In a particular embodiment, 0.5 1·5, 2, 2·5, 3, 3.5, 4, 4.5 5.5 20 7 ν, υ, 6.5, 8.5, 9, 9.5 or 1 G mg/m 2 per week. As disclosed herein (CN_P or related compounds. In another particular embodiment, each Γ: 1, 5, 15, 20, 25, 3, 35, 4, 5. ^, 90, or 100 mg of the platinum compound. The compound can be administered in a single large dose in a short period of time in the examples of the present invention, and it is disclosed here that, for example, about 〜。。。。。。。。。。。。。。。。。。。。 , I large dose - 』 /, thyroid and other compounds 49 200831525 through a continuous infusion for at least 24, 48, 72, 96 or 120 hours at the same time. In the fang dry example, through continuous injection or high dose injection The administration of the triclinide compound and/or one or more of the indicating compounds can be repeated at a frequency that is at least once a week, once every two weeks, once every three weeks, once every month, every five times, every five times. Once a week, once every six weeks, every eight weeks, every ten times, and/or every twelve weeks. Type of drug and frequency The administration cycle can be combined in any of the ways described herein. The triclinide compound and/or one or more platinum compounds can be administered repeatedly through a certain administration cycle, for example, in a large dose injection every two weeks for three times. Month. The Dosing 10 program can be administered for at least one, two, three, four, five, six, seven, eight, nine, one, eleven, eleven, eight, eight, or four months. At least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15 or 20 administration cycles. The triclinide compound and / or one or more compounds can be disclosed according to the disclosure herein. A combination administration, such as a triplicidin compound and/or one or more platinum compounds, can be administered once a week for three cycles every three weeks. In other embodiments, the compound can be administered at least once a day for a duration. At least 2, 3, 4, 5, 6, 7, 8, 9, or 1. This administration is followed by a corresponding cycle time for the unadministered triclinide compound and/or one or more platinum compounds. TCN, TCN-P and related compounds and one or more platinum compounds as disclosed herein can effectively cause tumors The amount administered is administered to the patient. Administration of TCN, TCN-P and related compounds and one or more platinum compounds can be provided in at least a partial response of at least 15-20% of the individual in vivo, such as at least 15%, 20% or 30. % reaction or complete reaction. In several embodiments, 50 200831525 at least 2, 5, 10, 15, 20, 30 or 50 mg/m 2 of the Qucilic compound disclosed herein can be applied to one body. In the example, at least 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, 1 〇, π, 15, 17, 20, 25, 30, 35, • 5 4〇, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 165, 175, 200, 250, 300 Or, 35 mg/m 2 as disclosed herein, TCN, TCN-P, TCN-PM or related compounds may be administered to one body. In several embodiments, 1 〇, 20, 50, 100, 150, 200, 250, φ 300, 35 〇 or 4 〇〇 mg of the platinum compound can be administered to one body. 10 The administration of the compound can be based on any of the treatment plans disclosed herein.

行。於特定實施例中，投藥計劃包括同時、循序或於一段時間投予低於約20毫克/平方米TCN及相關化合物及少於約 1〇〇毫克鉑化合物。於一個實施例中，低於2〇毫克/平方米 TCN或相關化合物可與少於約1〇〇毫克鉑化合物同時每週 15 一次投予。於另一個實施例中，低於約20毫克/平方米TCN 或相關化合物可每週一次投予，而少於約100毫克鉑化合物 ^ 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、1〇耄克/平方米及/或15毫克/平方米TCN或相關化合物及少於 20約30、25、20、50或100毫克鉑化合物可投予一個體。於另一個實施例中，少於10毫克/平方米曲西立濱化合物及少於約100*克鉑化合物可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髮、頻率、週期數及劑量之任一種組合。 51 200831525 5.5病人族群之篩檢於本發明之另一個態樣中，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。於一個實施例中，提供於哺乳動物治療癌症或腫瘤之方法，係 5經由⑴由該腫瘤獲得生物樣本；（ii)判定該癌症或腫瘤是否過度表現Akt激酶或南度活化之且填酸化之Akt激酶，以及 (iii)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。於一個實施例中，生物樣本可為活體組織檢查。於其它實施例中，生物樣本可為得自該腫瘤或癌症之流體、 10 細胞及/或抽吸物。生物樣本可根據熟諳技藝人士已知之任一種技術獲得。於一個實施例中，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，作有其它 15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（_根細長薄管於末端有密切對焦望遠鏡供觀看）穿過自然身體孔口^亦 20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，俾便獲得小旦組織供研究。内視鏡手術係依據欲觀看及/或欲治療之哭官或身體區域來命名。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）、月勺 200831525 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。此類型活體組織檢查可由胸骨或髂脊髖骨（後背骨盆兩侧的骨區） 5 施行。清潔皮膚，給予局部麻醉劑來麻痒該區。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。於另一實施例中，切除或切開活體組織檢查可於哺乳 10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（使用手術縫線來缝合關閉）。當整個腫瘤被移除時，稱作為切除活體組織檢查技術。當只移除部分腫瘤時’稱作為切開活體組織檢查技術。例如當懷疑為黑素 15瘤（一類皮膚癌）時經常以切除活體組織檢查為佳。又有其它實施例中，可使用細針抽吸(FNA)活體組織檢查。此型活體組織檢查使用一根細針來由腫瘤移出極小瑰組織。偶爾使用局部麻醉來麻痺該區，但本程序罕見造成重大不適且不留疤。例如FNA並未用於可疑的痣的檢查， 2〇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）玎用來將細針導引入内臟器官諸如肺臟或肝臟的腫瘤。於其它實施例中，可進行衝孔、剃除及/或皮膚活體錐、織檢查。衝孔活體組織檢查涉及使用生檢儀器取出一段短 53 200831525 圓柱體或稱作「蘋果組織來取麟皮膚樣本。投予局部麻醉後，儀器於皮膚表面上旋轉至儀器穿過各層，真皮、表皮、和皮下的最表淺部分（皮下脂肪）。剃除活體組、我榀查涉及藉刮除來移出皮膚的頂 5 10 15 層。剃除活體組織檢查也係Μ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。活體組織才欢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激酶之方法。Akt激酶之過度表現可參照激酶之磷酸化狀態。Akt 之回度磷酸化可根據此處所述方法檢測。於一個實施例中i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填酸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激酶碟酸化，諸如比對照組織所含之尬激酶至少約 h5、2、2.25、2.5、2.75、3、3·25、3·5、3·75、 4、4.25、4.5 '4 75、< ^ ^ r n 0 。 5、5·5、6、7、8、9、或1〇倍大量之 Akt激酶。於一個實施例中，本發明提供一種於個體或於 2〇個體之生物樣本中檢測失序的Akt激梅表現之方法，嗲、Λ 係經由讓得自該個體之細胞、細胞萃取物、血清或二= 本或讓該生物樣本與對該Akt激酶或其抗原部分為特= 之免疫交互作用分子接觸，以及篩檢免疫交互作用= -Akt激酶複體的形成程度，其中複體存在量相對於正^ 54 200831525 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。於另一個實施例中，經由篩檢編碼Akt激酶基因之表現 5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可得自個體之細胞或得自個體之生物樣本，視需要可產生cDNA。然後 10 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。本發明之又另一個實施例涵蓋於定量或半定量診斷套 15 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。試劑可包括例如洗滌緩衝液、抗體稀釋緩衝液、阻斷緩衝液、 20 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 55 200831525Row. In a particular embodiment, the dosing schedule comprises administering less than about 20 mg/m2 of TCN and related compounds and less than about 1 mg of platinum compound simultaneously, sequentially or over a period of time. In one embodiment, less than 2 mg/m2 of TCN or related compound can be administered once a week with less than about 1 mg of platinum compound. In another embodiment, TCN or related compounds below about 20 mg/m 2 can be administered once a week, while less than about 100 mg of platinum compound ^ can be administered next week. In additional embodiments, 2 mg/m 2 , 5 mg/m 2 , 1 g/m 2 and/or 15 mg/m 2 of TCN or related compounds and less than 20 about 30, 25, 20, 50 or 100 mg of platinum compound can be administered to one body. In another embodiment, less than 10 mg/m2 of tricineridine compound and less than about 100*gram of platinum compound can be administered to a body for at least 5 days through continuous infusion. The present invention provides any combination of administration, frequency, number of cycles, and dosage as disclosed herein. 51 200831525 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 one embodiment, a method of treating cancer or a tumor in a mammal is provided by (1) obtaining a biological sample from the tumor via (1); (ii) determining whether the cancer or tumor overexpresses Akt kinase or Southern activation and is acidified. Akt kinase, and (iii) treating the cancer or tumor with tripceine 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, 10 cells, and/or aspirate obtained 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 clinic, and other 15 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 performed through a fiberoptic endoscope (a thin tube with a close-focus telescope at the end for viewing) through a natural body orifice (also known as a rectum) or a small incision (ie an arthroscope). Check) to proceed. Endoscopy is used to see if the problematic organ has an abnormal or suspicious area, and the small Dan tissue is available for research. Endoscopic surgery is named after the crying officer or body area that is to be viewed and/or to be treated. The physician inserts the endoscope into the gastrointestinal tract (digestive tractoscopy bladder (cytoscopic examination), abdominal cavity (laparoscopic examination), joint cavity (arthroscopy), lunar 200831525 middle cavity (longitudinal examination), or tracheal and Bronchial system (pharyngoscopy and bronchoscopy). In another embodiment, a bone marrow biopsy is performed. This type of biopsy can be performed by the sternum or sacral hip bone (the bone area on both sides of the back pelvis). The skin is given a local anesthetic to itch the area. A slender, 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 be followed by aspiration Procedure. In another embodiment, excisional or incisional biopsy can be performed on 10 animals. This type of biopsy is often used when a wider or deeper skin portion is required. Use a scalpel to remove the full skin thickness. Further examination, and suturing the wound (using surgical sutures to suture off). When the entire tumor is removed, it is called excisional biopsy When only a part of the tumor is removed, it is called a biopsy technique. For example, when it is suspected that it is a melanoma 15 (a type of skin cancer), it is better to use a biopsy. In other embodiments, fine Needle aspiration (FNA) biopsy. This type of biopsy uses a fine needle to remove very small rose tissue from the tumor. Occasionally local anesthesia is used to paralyze the area, but this procedure rarely causes significant discomfort and does not leave any defects. FNA is not used for suspicious sputum examinations, but can be used to examine large lymph nodes near melanoma to understand whether melanoma has metastasized (diffusion). Computerized tomography (CT scan or CAT scan) is used. Introducing fine needles into tumors of internal organs such as the lungs or liver. In other embodiments, punching, shaving, and/or skin cone and woven examination may be performed. The punch biopsy involves taking a section using a biopsy instrument. Short 53 200831525 Cylinder or "Apple tissue to take a skin sample. After local anesthesia, the instrument rotates on the surface of the skin until the instrument passes through the layers, the dermis, The epidermis, and the superficial part of the skin (subcutaneous fat). Shaving the living group, I investigated the top 5 10 15 layers that were removed by scraping to remove the skin. The shaving biopsy was also performed by local anesthesia. Tissue examination involves taking a skin sample for microscopic examination to determine the presence of melanoma. The living tissue is only under local anesthesia. In a specific embodiment, a method for determining whether a tumor overexpresses Akt kinase is provided. Akt kinase The overexpression can be referenced to the phosphorylation state of the kinase. The phosphorylation of Akt can be detected according to the methods described herein. In one embodiment, the i tumor biopsy can be compared to the control tissue. The control tissue can be from Normal tissue of a mammal in a biopsy, or normal tissue obtained from a healthy mammal. Akt kinase overexpression or high acid filling can be determined that the tumor biopsy contains a greater amount of Akt kinase and/or Akt kinase acidification than the control tissue. , such as at least about h5, 2, 2.25, 2.5, 2.75, 3, 3, 25, 3. 5 than the 尬 kinase contained in the control tissue. 3 · 75, 4,4.25,4.5 '4 75, < ^ ^ r n 0. 5, 5, 5, 6, 7, 8, 9, or 1 〇 a large amount of Akt kinase. In one embodiment, the invention provides a method of detecting a disordered Akt-induced plum expression in an individual or in a biological sample of a 2 〇 individual, the sputum, sputum, by allowing cells derived from the individual, cell extracts, serum or II = or the biological sample is contacted with an immune interaction molecule that is specific for the Akt kinase or antigenic portion thereof, and the degree of formation of the immune interaction = -Akt kinase complex, wherein the amount of the complex is relative to Positive ^ 54 200831525 Cell elevation, indicating a disordered cell 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 in the cell is detected at the gene level by screening for 5 degrees of expression of the Akt kinase gene, 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 a cell of an individual or a biological sample obtained from an individual, and cDNA can be produced as needed. The 10 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 can be evaluated Or whether the concentration of cDNA is higher than that of the normal control group. Still another embodiment of the present invention encompasses antibodies against Akt kinase in a set of 15 quantitative or semi-quantitative diagnostic kits, including monoclonal antibodies or polyclonal antibodies for determining Akt kinase in suspicious cancer cells 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. The reagent may include, for example, a washing buffer, an antibody dilution buffer, a blocking buffer, a 20-cell staining solution, a developing solution, a stopping solution, an anti-phospho-protein-specific antibody, an anti-ubiquitin-specific antibody, a secondary antibody, and distilled water. . The kit group also includes instructions for use, either automated or semi-automated, or in a form compatible with automated machines or software. In one embodiment, the phosphoric acid of the AKT activated form (Akt phosphorylated at serine 474) is detected -ser-473 55 200831525

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: p.2324-2330。 5·6 Akt激梅Akt antibodies can be used as antibodies in the diagnostic kit set. See, for example, Yuan et al., Oncogene, 2000. 19: p. 2324-2330. 5·6 Akt 激梅

Akt也定名為PKB3，表示絲胺酸/蘇胺酸激酶之一個亞 5 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Thr3G8的磷酸化及於C端活化功能部位的 Ser473。Akt藉PTEN腫瘤阻遏子負向調節。PTEN之突變已 10 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 Nicosia S.V·，於癌症百科參考，2001，Schwab D.(編輯） 15 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活性之抑制於嗔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 Nicosia S.V·，於癌症百科參考，2001，Schwab D·(編輯） 20 Springer，柏林、海德堡及紐約；35-37頁；Sim 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之活化顯示與腫瘤的入侵及化學品抗性相關（West Κ·Α·等人’藥物Resist Updat，2002· 5: ρ·234-248)。 200831525Akt is also named PKB3, representing a subfamily of serine/threonine kinases. Three members AKT1, AKT2, and AKT3 have been identified in this subfamily. Akt is activated by extracellular stimulation 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 Thr3G8 in the activation loop and activation of the functional site of Ser473 at the C-terminus. 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 malignant diseases (Datta SR et al., Gene Dev, 1999. 13: ρ·2905-2927; Cheng JQ· and Nicosia SV·, Cancer Encyclopedia Reference, 2001, Schwab D. (ed.) 15 Springer, Berlin, Heidelberg and New York; 35_37). Akt is specifically responsible for the ectopic expression of active Akt, which induces cell survival and malignant transformation; whereas Akt activity inhibits apoptosis in the range of mammalian cells (Datta S.R_ et al., Gene Dev, 1999· 13: ρ·2905-2927 ; Cheng JQ· and Nicosia SV·, Reference for Cancer Encyclopedia, 2001, Schwab D· (eds.) 20 Springer, Berlin, Heidelberg and New York; 35-37; Sim Μ. et al., Am J Path, 2001. 159: ρ·431-437; Cheng JQ· et al., Oncogene, 1997. 14: p. 2793-2801). In addition, activation of Akt has been shown to be associated with tumor invasion and chemical resistance (West Κ·Α· et al. Drug Resist Updat, 2002·5: ρ·234-248). 200831525

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 extent 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、施2、Akt3。人Aktl、Akt2、及The Akt kinase can be any of the known Akt family kinases, or its related kinases 'including but not limited to Aktl, Shi 2, Akt3. Human Aktl, Akt2, and

Akt3之mRNA及胺基酸序列分別顯示於第以气、乃^及8a_c 圖0The mRNA and amino acid sequences of Akt3 are shown in the first gas, the gas, and the 8a_c, respectively.

5·7·診斷檢定分析 1〇免疫檢定分析、於-個實施例中，提供-種於哺乳動物細胞或於得自哺礼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其它樣本或生物樣本與對Akt激崎或其抗原部分具有特異 15性之免疫交互作用分子接觸；以及筛檢免疫交互作用分子5.7. Diagnostic assay analysis 1 〇 immunoassay analysis, in one embodiment, provides a method for detecting the disordered expression of Akt kinase in a mammalian cell or in a biological sample obtained from a feeding animal, the method being Exposing cells, cell extracts or serum or other samples or biological samples obtained from the mammal to molecules that are immunologically interacting with Akt akisaki or its antigenic portion; and screening for immunological interaction molecules

Akt激酶複體形成程度；以及狀是否存在有相對於正常細胞複體存在量之升高。 20 疫交互作用 > 子可為對Akt激酶或其抗原部分或盆同系物或触物具有特異性及結合親和力個實施例中’免疫交互翻分何為免疫球蛋自分子。於其它實施例中，免疫交互_分子可為抗㈣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 (ΤΑΒΜ)。於-特定實施例中，抗體可為單株抗體。於另— 特定實施财，抗體可為多株抗體。免疫交互仙分子對 57 200831525The extent of Akt kinase complex formation; and whether there is an increase in the presence of a normal cell complex. 20 Quarantine interactions > Sub-specificity and binding affinity for Akt kinase or its antigenic portion or potent homologue or touch. In an example, the immunological interaction is an immunoglobulin self-molecule. In other embodiments, the immunological interaction molecule can be an anti-(four) segment, a single chain antibody, and/or a de-immunized molecule comprising a humanized antibody and an antigen binding molecule associated tau cell (ΤΑΒΜ). In a particular embodiment, the antibody can be a monoclonal antibody. In another case, the antibody may be a plurality of antibodies. Immune interaction fairy pair 57 200831525

Akt激峰有特異性，或更特別對Akt激酶上的抗原定子或抗原決定部位有特異性。Akt激酶上的抗歧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部位可為B細胞抗原決定部位，或若屬適細胞抗原決 5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、、二由讓彳于自该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 1〇體或其上之抗原定子或抗原決定部位接觸；以及然後定量或疋性測定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備。例如用於人Akt激酶之檢測，抗體通常但非必要衍生自 15 非人動物諸如靈長類、牲口動物(例如羊、牛、豬、山羊、馬）、貫驗室試驗動物(例如小鼠、大鼠、天竺鼠、兔)及/或伴倍動物（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於試管内於細胞或組織之生檢進行。但若抗體經過適當脫 2 〇 > 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激酶抗體可為癌症把定劑。如此’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。 58 200831525 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#又製造則可來自於細胞培養。Akt激酶可藉任一種適當手段而分離自生物樣本。例如分離可利用Akt激酶表面 5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成：超離心、離子交換層析術(例如陰離子交換層析術、陽 10離子交換層析術）、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、免疫沉澱）。Akt 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 15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之任種彳示準協疋方案來進行，例如說明於Kohler及 20 Milstein (Kohler及Milstein，自然，1975· 256: ρ·495-499 ; Kohler及Milstein，Eur J Immimol，1976. 6(7): p.511-519);Akt peaks are specific, or more specifically specific for antigenic stator or antigenic epitopes on Akt kinase. The anti-alias or epitope on the Akt kinase includes the molecular portion to which the immune response is directed. The antigenic stator or antigenic epitope can be a B cell epitope, or a suitable cell antigen. In one embodiment, the antibody is a phospho-ser 473 Akt antibody. One embodiment of the present invention provides a method of developing a cancer growth or a series of cancer growth in a mammal in which disordered Akt activity is present, which is derived from or derived from a cell or cell extract of the mammal. The biological sample of the individual binds to an Akt kinase-effective amount of an Akt kinase-specific anti-steroid or an antigenic stator or epitope thereof; and then quantitatively or sputumly determines the content of the Akt kinase-antibody complex, wherein It was judged that the complex had a higher content than normal cells. 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 kinase, antibodies are usually, but not necessarily, derived from 15 non-human animals such as primates, livestock animals (eg sheep, cattle, pigs, goats, horses), laboratory test animals (eg mice, Rats, guinea pigs, rabbits) and/or accompanying animals (eg dogs, cats). Antibodies can also be produced recombinantly in prokaryotic or eukaryotic host cells. In general, antibody-based assays can be performed in vitro in a test tube or tissue. However, if the antibody is subjected to humanization by appropriate depurination and/or for human use, the antibody can be labeled, for example, by nuclear labeling, and administered to a patient to determine the position of the nuclear marker by radioactive techniques. Akt kinase antibodies can be used as a fixative for cancer. Thus, another embodiment of the present invention provides a deimmunized form of an antibody for cancer imaging for use in humans and non-human patients. 58 200831525 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 hand #. Akt kinase can be isolated from biological samples by any suitable means. For example, isolation may utilize any of the Akt kinase surface 5 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) or More than one. 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, positive 10 ion exchange chromatography) , 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 Dynabead ) (trade name) separation 'immunochromatography, immunoprecipitation). The separation of Akt kinase from biological fluids preserves the conformational epitopes present on the kinase, so that techniques for enzymatic denaturation are 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 individual antibodies can be carried out using any of the techniques known to the artisan, such as those described in Kohler and 20 Milstein (Kohler and Milstein, Nature, 1975 256: ρ·495-499; Kohler And Milstein, Eur J Immimol, 1976. 6(7): p. 511-519);

Coligan等人（免疫學之流行方案，1991_1997，約翰威利父子公司）或Toyama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 59 200831525 分或Akt激酶之重組形式藉標準方法免疫接種，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。於若干實施例中，Akt激酶片段可用來產生抗體。該片段可 5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法進行。例如經由使用EB病毒(EBV)之轉形方法可達成永 10生化(Kozbor等人，酶學方法，ΐ986·121:ρ·140)。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，1991-1997，參見上文），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體製造性體細胞，特別為B細胞係與骨髓瘤細胞系融合。此 15 等體細胞可衍生自二次感染動物較佳為齧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髓瘤細胞來用於融合瘤製造之融合程序（Kohler及Milstein， 20 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63-Ag8、P3乘63-AG8.653、P3/NSl-Ag4-l (NS-1)、 Sp2/0_Agl4及S194/5.XXO.Bu.l。P3乘63-Ag8及NS-1 細胞系 200831525 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 ^ 1978· 148(1)·· ρ·313-323)。產生抗體製造性脾細胞或淋巴結 • 5 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，將體細胞與骨二髓瘤細胞以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 _ 文；Kohler及Milstein，1976，參見上文；Gefter等人，體 10細胞遺傳學，1977· 3: ρ·231·236 ; Volk等人，1982，來見上文）。該等研究學者所使用之融合促進劑為山戴病毒(Sendai vims)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交體之手段。大致上，融合細胞雜交體之選擇伴隨有於培 _ 15養基中培養細胞，該培養基可支援融合瘤的生長，但阻止切合骨_細胞之生長，未融合骨_細胞通常會繼續馨減***。融合所使用之體細胞於試管内培養並未保有長期存活力，如此不會成問題。需要數週來選擇性培養融: 細胞雜交體。於本時間早期，需要識別可製造期望之抗體 20之該等雜交體，讓其隨後可被轉殖及繁殖。大致上，約10% 所得雜錢可製造《之㈣，㈣1%至觸％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析方法中之任-種達成，該等方法包括_結免疫檢定^ 析技術及放射性免疫檢定分析技術，例如說明於 61 200831525 人(單株抗體及融合瘤：生物分析之新紀元，[980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)。一旦已經選擇期望之融合細胞雜交體且轉殖入個別抗 — 5 體製造性細胞系，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濃度製造單株抗體。另外，個別細胞系可於試管内於實驗春 10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Akt 激酶之特異性。舉例言之，細胞系可分配入多個孔内及培養，來自各孔之上清液藉酶聯結免疫吸附檢定分析 15 (ELISA)、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弟二過識別，然後於試管内直接培養，或注射入組織可相容馨動物體内來形成腫瘤，以及製造、㈣、及純化㈣抗體。因此，本發明提供-種於-樣本中檢測Akt激酶或其片 20段、、欠異株或街生物之方法，包含該樣本與抗體或其片段或其何生物接觸；檢測比較正常對照組，含有該抗體及塵激酶或其片段變異株或衍生物之複體之含量，其中測以匕激酶之升呵私度。任何測定複體之形成之適當技術皆可使用舉例σ之’根據本發明具有結合之通報子分子之抗體 62 200831525 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 (ELISA)、免疫層析技術(ICT)及西方墨點，此等免卢_定競爭分析為熟諳技藝人士眾所周知。免疫檢定分析也包括 5 檢定分析。本發明涵蓋定性及定量免疫檢定分_。、π罔寻利茶 4,016,043 ; 4,424,279 ·，及4,〇18,653。此等技術包括非競爭型單位置及二位置檢定分析及傳統競爭結合檢定分析。此Coligan et al. (Popular Program for Immunology, 1991_1997, John Wiley & Sons) or Toyama et al. (Handbook for Individual Antibody Experiments, 1987, Science Lecture). In general, an animal is immunized by a standard method using a biological fluid containing Akt kinase or a recombinant form thereof, 2008 20082525 or Akt kinase to produce an antibody-producing cell, particularly an antibody-producing somatic cell (eg, B lymphocyte). ). 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 5 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, by using the EB virus (EBV) transformation method, the chemistry can be achieved (Kozbor et al., Enzymology Method, ΐ 986.121: ρ·140). In another embodiment, antibody-producing cell lines are immortalized using cell fusion methods (described in Coligan et al., 1991-1997, supra), and cell fusion methods are widely used to produce monoclonal antibodies. In such a method, an antibody-producing somatic cell having the potential to produce an antibody, particularly a B cell line, is fused to a myeloma cell line. The somatic cells can be derived from lymph nodes, spleen and peripheral blood of secondary infected animals, preferably rodents such as mice and rats. In a particular embodiment, mouse spleen cells can be used. In other embodiments, rat, rabbit, sheep or goat cells can also be used. Specialized myeloma cells have been developed from lymphocytic tumors for use in fusion procedures for fusion tumor manufacturing (Kohler and Milstein, 20 1976, supra; 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 63-Ag8, P3 by 63-AG8.653, P3/NSl-Ag4-l (NS-1), Sp2/0_Agl4, and S194/ 5.XXO.Bu.l. The P3 by 63-Ag8 and NS-1 cell lines 200831525 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 was reported by Trowbridge (j Exp Med ^ 1978. 148(1)·· ρ·313-323). Production of antibody-producing splenocytes or lymph nodes • The method of hybridization of 5 cells with myeloma cells is usually carried out in the presence of an agent (chemical, viral or electrical) that promotes fusion of cell membranes, and somatic cells and osteomyeloma cells. 10:1 ratio 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); Kohler and Milstein, 1976, supra; Gefter et al, Body 10 cytogenetics, 1977. 3: ρ·231.236; Volk et al. 1982, see above). The fusion promoters used by these researchers are Sendai vims and polyethylene glycol (PEG). In several embodiments, a means of selecting a fused cell hybrid 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 culture, which supports the growth of fusion tumors, but prevents the growth of osteoblasts, and unfused bones usually continue to divide and divide. . In vivo culture of 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 fused: cell hybrid. Early in the day, it is necessary to identify such hybrids that produce the desired antibody 20, which can then be subsequently propagated and propagated. In general, about 10% of the resulting miscellaneous money can be made in the range of (4), (4) 1% to touch % and ° is not uncommon. The detection of antibody-producing hybrids can be achieved by any of several standard assay methods, including _ knot immunoassay techniques and radioimmunoassay analysis techniques, for example, as described in 61 200831525 Human (monoclonal antibody) And fusion tumors: a new era of bioanalysis, [980, Plenum Press, New York, pp. 376-384); and can be reached by FAcs analysis (O'Reilly et al., Biotechnology, 1998 25: P_824-830). Once the desired fused cell hybrid has been selected and transfected into individual anti-5 constructive cell lines, 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 animal that was injected was subsequently developed to secrete a specific monoclonal antibody produced by the fusion cell hybrid. Animal body fluids such as serum or ascites can be tapped to produce monoclonal antibodies at high concentrations. Alternatively, individual cell lines can be propagated in test tubes in a test chamber in a 10-room 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 the specificity of the Akt kinase of interest. For example, the cell line can be distributed into a plurality of wells and cultured, and the supernatant from each well is analyzed by enzyme-linked immunosorbent assay 15 (ELISA) and indirect fluorescent antibody technology. The identifiable target LIM kinase - but does not recognize the non-target epitopes of the individual antibody-producing cell lines, and then directly cultured in a test tube, or injected into a tissue-compatible succulent animal to form Tumors, as well as manufacturing, (d), and purified (iv) antibodies. Accordingly, the present invention provides a method for detecting Akt kinase or a fragment thereof, a septic strain or a street organism in a sample, comprising contacting the sample with an antibody or a fragment thereof or a living organism thereof; and detecting a normal control group, The content of the complex containing the antibody and the dust kinase or a fragment variant or derivative thereof, wherein the concentration of the lumbrokinase is measured. Any suitable technique for determining the formation of a complex can be used for the immunoassay analysis of an antibody having a combined reporter molecule according to the present invention. Such immunoassay analysis includes, but is not limited to, radioimmunoassay analysis (RIA), enzyme-linked immunosorbent assay (ELISA), immunochromatography (ICT), and Western blotting. People are well known. The immunoassay analysis also includes 5 assay analyses. The present invention encompasses qualitative and quantitative immunoassay scores. π罔寻利茶 4,016,043 ; 4,424,279 ·, and 4,〇18,653. These technologies include non-competitive single-position and two-position calibration analysis and traditional competitive combination verification analysis. this

10 抗原等檢定分析包括經過標記之抗原結合分子直接結合至標輕抗原。本發明進-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ώ晰口变曰貝表現程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糸統來定ΐ Akt蛋白質表現或活化程度之方法。成 15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行染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Ακτ蛋白質表現量或活化私度。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AKT1&AKT2蛋白 20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AKT蛋白質數量。可使用活化特徵特異性試劑來對活化AKT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AKT數量及AKT活化狀態的變化。 63 200831525 於本發明方法之一個特定實施例中，於細胞或組織樣本中之AKT蛋白貝表現程度可使用酶聯結免疫吸附檢定分析(ELISA)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2002/0015974。 5 於其它實施例中，酶免疫檢定分析可用來檢測Akt激 - 酶。於此種檢定分析中，酶軛合至第二抗體，通常係利用戊二醛或過碘酸鹽來軛合至第二抗體。用於特定酶之酶基二質通常係於被相對應之酶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，而非馨 10使用產色酶基質。經過酶標記之抗體可添加至第一抗體-抗原複體，允許其結合，然後洗滌去除過量試劑。含有適當酶基質之溶液隨後添加至抗體-抗原-抗體複體。酶基質與聯結至第二抗體之酶反應，產生定量視覺信號，該信號通常可藉分光光度辦法進一步定量，來獲得樣本中所存在之抗 15原量之指示。另外，螢光化合物諸如螢光素、若丹明、及 — 鑭系化合物、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，經過螢光路桿記之翁抗體吸收光能，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。經過勞光標 20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光。觀察得螢光指示存在有感興趣之抗原。免疫鸯光計量檢定分析(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 64 200831525 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_特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激酶抗體制動於固體撐體，且讓其與 • 5 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之任一種檢測，全部變化皆涵蓋於本發明且為熟諳技藝人瞻士已知。此項辦法允許例如使用基於血清之檢定分析來快 10 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 Bioscience)之 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。elisA 15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、抗體稀釋缓衝液、阻斷緩衝液、細胞染色 ^ 溶液、展開溶液、停止溶液、二次抗體及蒸餾水。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 20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熟諳技藝人士已知之任一種適當技術測定。於一個實施例中，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 65 200831525 之多核苷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募核苷酸引子協同用於核酸擴增反應，諸如尺丁 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（1991，科學251:767-777)及Kazal 5等人（1996，自然藥物2:753-759)。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，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TRIZOL試劑(GIBCO-BRL/生命科學，馬里蘭州 10 蓋色堡）分離。寡-dT或隨機序列寡核苔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反應中以特定序列寡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 15 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 20 致上參考Mullis等人（1987, Quant Biol 51:263; Erlich (編輯），PCR技術，1989，史塔克敦出版社，紐約）。如此，藉 PCR擴增特定核酸序列仰賴具有保留的核苔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 66 200831525 * 5 • 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 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可使用與特定募核苷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 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 10 獻Lawn等人，1981，核酸研究2:6103; Goeddel等人，1980，核酸研究8:4057);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（Invitrogen))之轉殖位置；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。 15 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、相鄰内襯及髮夾寡探針及自行發螢光擴增基 20 因。此等化學及即時PCR大致上係討論於Mackay等人， 2002，核酸研究30:1292-1305; Walker，2001，J Biochem Mol 毒理學 15:121-127; Lewis等人，2001，J Pathol 195:66-71。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答酸序列與具有選自於 67 200831525 第6a-c、7a-d、或8a_c圖之募核苷酸序列之Akt序列或其片 4又互補之序列之养核％•酸棟針接觸；以及然後經由該探針/、序列雜父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 5探針而檢測。探針例如可以放射性同位素，或以可取得抗體之生物素、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 10核苷酸限剪技術、點打點檢定分析、反點打點檢定分析、線探針檢定分析及5’磷酸酶檢定分析。另外，探針可使用一：!又適用之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父至選自於第6a_c、7a-d及8a-c圖之核苷酸或其片段 15之約14、15、16、18、20、25或28核苔酸。通常使用長度大於約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10 Antigen and other assays include direct binding of labeled antigen-binding molecules to standard light antigens. The present invention further provides a method for quantifying the degree of expression and degree of activation of a mussel in an automated subject such as a cancer patient or a cell or tissue sample suspected of having a cancer. In one embodiment, the invention provides a method of quantifying the extent of Akt protein expression or activation using an imaging system. The 15-image system can be used to receive, enhance, and process images of cell samples or tissue samples that have been stained with AKT protein-specific staining, and to determine the amount of Ακτ protein expression in cells or tissue samples obtained from such animals or Activate privacy. In one embodiment of the methods of the invention, a calibration curve for AKT1 & AKT2 protein 20 performance 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 AKT protein expressed in the cell or tissue sample. A similar calibration curve can be made for the activated AKT protein using an activation signature specific reagent. It can also be used to determine changes in the number of AKTs and AKT activation status before and after clinical cancer treatment. 63 200831525 In a particular embodiment of the method of the invention, the degree of expression of the AKT protein in a cell or tissue sample can be quantified using enzyme-linked immunosorbent assay (ELISA) to determine the AKT protein content in the sample. Such a method is described, for example, in U.S. Patent Publication No. 2002/0015974. 5 In other embodiments, an enzyme immunoassay assay can be used to detect an Akt-enzyme. In this assay, the enzyme is conjugated to a second antibody, typically conjugated to a second antibody using glutaraldehyde or periodate. The enzyme base used for a particular enzyme is typically selected to produce a detectable color change upon hydrolysis by the corresponding enzyme. It is also possible to use a fluorogenic enzyme substrate that obtains a fluorescent product, rather than a chromogenic enzyme substrate. The enzyme-labeled antibody 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 an enzyme that binds to the second antibody to produce a quantitative visual signal that can be further quantified by spectrophotometry to obtain an indication of the amount of anti-15 original present in the sample. In addition, fluorescent compounds such as luciferin, rhodamine, and lanthanide compounds, ruthenium (EU) can be chemically coupled to the antibody without changing the binding energy. When activated by illumination of a specific wavelength of light, the fluorescent light absorbs the 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 was recorded to the first antibody-antigen complex by a 20-fold antibody. After washing to remove unbound reactants, the remaining ternary complexes are exposed to 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, chemistry 64 200831525 luminescent molecules or bioluminescent molecules may also be employed. In particular embodiments, antibodies to Akt kinase can also be used in Akt kinase-specific sera or other circulating body fluids for detection of Akt kinase by ELISA. This can be achieved by immobilizing the anti-Akt kinase antibody to a solid support 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 the 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 group of signaling cell activation ELISA kits for Akt S473 from SuperArray Bioscience can be used in the present invention. In one embodiment, the antibody can be an anti-pan-antibody that recognizes Akt S473. The elisA 15 assay suite contains anti-Akt antibodies and additional reagents including, but not limited to, wash buffer, antibody dilution buffer, blocking buffer, cell staining solution, developing solution, stop solution, secondary antibody, and distilled water. Nucleotide Detection In another embodiment, a method of detecting Akt kinase by detecting the extent of expression of a polynuclear glycoside encoding an 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. In one embodiment, 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 in conjunction with a message sequence and an anti-message sequence encoding a polynucleotide of the kinase 65 200831525 or a nucleotide primer corresponding to the collateral sequence therein. Nucleic acid amplification reactions, such as ruler PCR. A variety of automated solid phase detection techniques are also available to those skilled in the art, such as those described by Fodor et al. (1991, Science 251: 767-777) and Kazal 5 et al. (1996, Nature Medicine 2: 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, such as total RNA isolated from human cancer tissue. RNA can be isolated by methods known in the art, for example using TRIZOL reagent (GIBCO-BRL/Life Sciences, Fortune, Maryland 10). Oligo-dT or random sequence oligonucleotides and specific sequence oligonucleotides can be used as primers in the reverse enzymatic reaction to prepare the first strand of cDNA from the isolated RNA. The resulting first strand of cDNA is then amplified in a PCR reaction with a specific sequence of oligonucleotide oxalate 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 acid, RNA and/or 15 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 sequence of such primers may be the same 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. The reference is directed to Mullis et al. (1987, Quant Biol 51:263; Erlich (ed.), PCR Technology, 1989, Starkton Press, New York). Thus, amplification of a particular nucleic acid sequence by PCR relies on an oligonucleotide or primer having a retained nucleotide sequence, which is presumed by the alignment of the relevant gene sequence or protein sequence, such as the sequence of a mammalian Akt kinase gene. Compare 66 200831525 * 5 • Presumption. 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 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 hybridization to a Southern blot of a specific nucleotide probe, or its electrophoretic activity can be compared to a dna standard having a 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, see Reference 10, Lawn et al., 1981, Nucleic Acids Research 2: 6103; Goeddel et al. , 1980, Nucleic Acids Research 8: 4057); the amplified product is transferred into a suitable vector such as the pCRII vector (Invitrogen); the inserted insert is sequenced; This DNA sequence is achieved by comparison with known sequences of LIM kinase. The relative amounts of LIM kinase mRNA and cDNA were 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 the possible translational activity of available mRNA transcripts. Real-time PCR programs and other PCR programs use a variety of PCR products for detection chemistry, including DNA-binding fluorophore binding, 5' endonuclease, adjacent lining and hairpin oligo probes, and self-fluorescent amplification 20 reasons. Such chemical and real-time PCRs are generally discussed in Mackay et al., 2002, Nucleic Acids Research 30: 1292-1305; Walker, 2001, J Biochem Mol Toxicology 15: 121-127; Lewis et al., 2001, J Pathol 195 :66-71. In another embodiment, the disordered performance of Akt can be identified by allowing a nucleic acid sequence isolated from a biological sample to have a map selected from 67a 3rd, 7a-d, or 8a_c selected from 67 200831525. The Akt sequence of the nucleotide sequence or the complementarity of the fragment of the fragment 4 is contacted with the nutrient core; and then the sequence is detected via the probe/, the sequence of the parent and comparing the result to the normal sample. Hybridization of the probe to the biological sample can be detected by labeling the 5 probe with any one of the detectable agents. The probe may, for example, be a radioisotope or be labeled with 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 oligo 10 nucleotide restriction techniques, dot-point assays, reverse dot assays, line probe assays, and 5' phosphatase assays. Alternatively, probes can be detected using any of the DNA array technologies, including macro array technology, microarray technology, and DNA microchip technology. Oligonucleotide oxalate probes typically include about 14, 15, 16, 18, 20, 25 or 28 nucleus moss from a parent to a nucleotide selected from the 6a-c, 7a-d, and 8a-c maps or fragment 15 thereof. acid. Probes having a length greater than about 25 or 28 nucleotides are generally 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 20 檢定分析測定。例如但非限制性，可使用Hogg等人（1994，致癌基因 9:98-96)，Mills 等人（1992，J Biol Chem 267:16000·006)及 Tomizawa 等人（2001，FEBS Lett 492:221-7)，Schmandt等人（1994, J Immunol 152:96-105)所述方法。其它絲胺酸、蘇胺酸及酪胺酸激酶檢定分析述於 200831525The activity of Akt kinase can be determined using any of the appropriate kinase 20 assay assays known to the art. For example and without limitation, Hogg et al. (1994, Oncogene 9: 98-96), Mills et al. (1992, J Biol Chem 267: 16000·006) and Tomizawa et al. (2001, FEBS Lett 492: 221) can be used. -7), the method described by Schmandt et al. (1994, J Immunol 152: 96-105). Other assays for serine, threonine and tyrosine kinase assays are described in 200831525

Ausubel等人（分子生物學之短協定方法，i999年單元 17.6)。施激酶檢定分析通常使用施多肽、經標記之施體蹲基質、及受體峰基質，言亥受體酶基質為Akt之特異性或非特 5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絲酶活性多肽可使用多種表 .現系統製造，可純化自細胞，可呈經裂解或未經裂解之重組融合蛋白質形式及/或可有非_施多肽序列例如仙標鐵 1〇或沒-半乳㈣崎於其N端或0端。若癌性細胞系用作為欲檢定分析之施的來源，驗t雜可於癌性、纟遗彳、巾檢定分析。適當Akt檢定分析用之施體酶基質包括對於藉旭去鱗酸化為敏感之任何分子，例如包括經r-標記之ATP及ATP 類似物其中該標記為33p、32p、35s或任何其它放射性同位 15素或適當螢光標記。Akt檢定分析用之適當接受者酶基質包括對於藉Akt㈤酸化為敏感之任—種多肽或其它分子。接受者酶基質係衍生自活體内旗標無片段。接受者酶基質片段之長度可為8至50胺基酸，通常為】〇至3〇胺基酸，特別長約 10 12 15、18、20及25胺基酸。其它接受者酶基質可於 20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—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，此處接受者酶基仏過生物耗，且透過其與麟S抗生物素之交互作用來、、屯化，或可取得特定抗體，該抗體可特異性識別接受者 69 200831525 酶基負。反應係於多種條件下進行，諸如於固體撐體上，於凝膠、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、或螢光攝影術測定所 _ 5 結合的放射性或螢光。 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、癌症及其它與異常細胞增生相關聯之病症之情況。於一個實施例中，本發明化合物可用於治療癌瘤、肉瘤、淋春 10 巴瘤、白血病、及/或骨髓瘤。於本發明之其它實施例中，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，諸如但非限於下列器官或組織之癌症：***、攝護腺、肺、支氣管、大腸、泌尿道、膀胱、非何杰金氏淋巴瘤、黑素瘤、腎、腎上腺、 15 胰、咽頭、甲狀腺、胃、腦、多發性骨髓瘤、食道、肝、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、小鲁腸、慢性骨髓性白血病、急性淋巴性白血病、肛門、肛管、肛門直腸、曱狀腺、陰門、膽囊、胸膜、眼、鼻、鼻腔、 2〇中耳、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、簟樣肉芽膜。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 70 200831525 瘤、血管内皮瘤、基底細胞癌、鱗狀細胞癌、及卡波西氏肉瘤，·及非惡性病或urn p ^ “ 病症諸如乾癖、淋巴血官新生、兒童血管瘤、杯广後姓^ 文σ ·知博氏（Sturge-Weber) _群、4疲、神經纖維瘤、結節性硬化症、產腹）牙腫、隱性營養不良性大泡性表 "κ m鲁 '表皮鬆解症、靜脈潰瘍、痤性角化症。 ^ 1雜肖化症、及光線Ausubel et al. (Short-term approach to molecular biology, i999 unit 17.6). The application of the kinase assay typically employs a polypeptide, a labeled donor matrix, and a receptor peak matrix, which is specific or non-specific for Akt. In this assay, Akt transfers the labeled moiety from the donor enzyme substrate to the receptor enzyme matrix, and the amount of the labeled enzyme is transferred to the receptor enzyme substrate by the number of labeled portions of the enzyme substrate. Table. Manufactured by the current system, purified from cells, may be in the form of a lysed or unlysed recombinant fusion protein and/or may have a non-application polypeptide sequence such as celestial iron 1 〇 or no-half milk (four) End or 0. If the cancerous cell line is used as the source of the assay to be assayed, the test can be analyzed on cancerous, sputum, and towel. Appropriate Akt assays for the application of the enzyme substrate include any molecule that is sensitive to de-squaring, including, for example, r-labeled ATP and ATP analogs wherein the label is 33p, 32p, 35s or any other radioisotope. Or appropriate fluorescent label. The appropriate recipient enzyme matrix for Akt assay analysis includes any polypeptide or other molecule that is susceptible to acidification by Akt (f). The recipient enzyme matrix is derived from the in vivo flag without fragments. The acceptor enzyme substrate fragment can be from 8 to 50 amino acids in length, typically from 〇 to 3 〇 amino acids, particularly about 10 12 15, 18, 20 and 25 amino acids. Other recipient enzyme matrices can be assayed using 20 different sets of polypeptides or other molecules. The TTK acceptor enzyme substrate can be purified from other reaction components after the reaction. This purification is usually carried out via molecular interactions, where the recipient's enzyme base is bio-depleted, and through its interaction with the s-S-biotin, it can be obtained, or it can be obtained, and the antibody can be specific. Sexual recognition recipient 69 200831525 Enzyme-based negative. The reaction is carried out under a variety of conditions, such as on a solid support, in a gel, 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, autoradiography, scintillation counting, scanning, or fluoroscopy to measure the radioactivity or fluorescence of the _5 binding. 6. Methods of Treatment The compounds and pharmaceutical compositions provided herein are useful in the treatment of conditions including swollen tumors, cancer, and other conditions associated with abnormal cell proliferation. In one embodiment, the compounds of the invention are useful in the treatment of carcinoma, sarcoma, lymphoma, leukemia, and/or myeloma. In other embodiments of the invention, the compounds disclosed herein are useful for treating solid tumors. The compounds of the invention are useful in the treatment of cancer, such as, but not limited to, cancers of the following organs or tissues: breast, prostate, lung, bronchi, large intestine, urinary tract, bladder, non-Hodgkin's lymphoma, melanoma, kidney, Adrenal gland, 15 pancreas, pharynx, thyroid, stomach, brain, multiple myeloma, esophagus, liver, intrahepatic bile duct, cervix, larynx, acute myeloid leukemia, chronic myelogenous leukemia, soft tissue such as heart, Hodgkin's lymph Tumor, testis, small intestine, chronic myeloid leukemia, acute lymphocytic leukemia, anal, anal canal, anorectal, sacral gland, vaginal canal, gallbladder, pleura, eye, nose, nasal cavity, middle ear, nasopharynx, Ureter, peritoneum, mesenteric, mesenteric, and gastrointestinal tract, high-grade gangeatoma, glioblastoma, large intestine, rectum, pancreas, gastric cancer, hepatocellular carcinoma; head and neck cancer, carcinoma; renal cell carcinoma; renal cancer; Sarcoma; hemangioendothelioma, lymphoma, leukemia, sputum-like granulosa. In additional embodiments, the compounds of the invention are useful in the treatment of dermatological conditions including, but not limited to, malignant disease vascular 70 200831525 tumor, hemangioendothelioma, basal cell carcinoma, squamous cell carcinoma, and Kaposi's sarcoma, and non-malignant diseases Or urn p ^ " Illness such as dryness, lymphocytic neonatal, hemangioma of the child, surname of the cup 广文σ · Sturge-Weber _ group, 4 fatigue, neurofibromatosis, tuberous sclerosis, Abdominal) edema, recessive dystrophic vesicular table " κ m Lu' epidermolysis, venous ulcer, spastic keratosis. ^ 1 schizophrenia, and light

10 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，包括本發日聽合物之組絲可㈣轉料㈣症及轉移癌症0 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1所列舉之癌症。表1 :癌症類別 ~〜^^--~---10 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. In addition, the composition of the present invention may include (4) transfusion (4) disease and metastatic cancer. 0 In other embodiments of the invention, the compounds described herein may be used to treat cancer, including but not limited to those listed in Table 1 below. Cancer. Table 1: Cancer Category ~~^^--~---

急性淋巴母細胞性白血病，成人急性淋巴母細胞性白血病，兒童急性骨髓性白血病，成人急性骨體性白血病，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’兒童骨癌髮狀細胞白血病頭頸癌肝細胞(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 71 200831525 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企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，參考蕈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5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，兒童多發性骨趙瘤/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，褥#Acute lymphoblastic leukemia, adult acute lymphoblastic leukemia, acute myeloid leukemia in children, acute osteomyelitis in adults, adrenal cortical carcinoma in children, adrenal cortical carcinoma, AIDS-related cancer, AIDS-associated lymphoma, anal cancer, astrocytoma , children's cerebellar stellate cell tumor, children's brain basal cell carcinoma cholangiocarcinoma, extrahepatic cholangiocarcinoma biliary 'children's bone cancer hair cell leukemia head and neck cancer liver cells (liver cancer), adult (primary) liver cells (liver cancer), Children (primary) Hodgkin's lymphoma 'Adult Hodgkin's lymphoma, Hodgkin's lymphoma in children, hypopharyngeal hypothalamic hypothyroidism and visual pathway neuroglioma, children's intraocular melanin Tumor pancreatic islet cell carcinoma (endocrine pancreas) Kaposi's sarcoma kidney (kidney cell) carcinoma Kidney cancer 'Children's laryngeal cancer laryngeal cancer, child 71 200831525 Osteosarcoma/malignant fibrous histiocytoma Brain stem neuroglioma, child brain Tumor, adult brain tumor, brain stem neuroglioma, childhood brain tumor, cerebellar stellate cell tumor, childhood brain tumor, brain astrocytoma / malignant Gelatinous, childhood brain tumor, ependymoma, childhood brain tumor, medulloblastoma, childhood brain tumor, undifferentiated neuroectodermal tumor on the cerebellum, childhood brain tumor, visual pathway and hypothalamus Tumor, childhood brain tumor, childhood breast cancer, childhood breast cancer, male bronchial adenoma/carcinoid, child Burkitt's lymphoma carcinoid, childhood carcinoid, gastrointestinal unknown primary central nervous system cancer Lymphoma, primary cerebellum, stellate cell tumor, childhood brain, astrocytoma/malignant glioma, childhood cervical cancer, childhood cancer, chronic lymphocytic leukemia, chronic myeloablative white disease, chronic myeloproliferative disorders, colorectal cancer Colorectal cancer, children's skin T-cell lymphoma, reference verrucous granuloma and Xie Saray syndrome endometrial cancer ependymoma, esophageal cancer esophageal cancer, Ewing's tumor leukemia, acute lymphoblastic , adult leukemia, acute lymphoblastic, childhood leukemia, acute myeloid, adult leukemia, acute myeloid, childhood leukemia, Lymphocytic leukemia, chronic myelogenous leukemia, B cell lip and oral cancer, adult (primary) liver cancer, childhood (primary) liver cancer, non-small cell liver cancer, small cell lymphoma, AIDS-associated lymphoma, Bergis's lymphoma, cutaneous T cells, reference to sputum granuloma and Sezary syndrome lymphoma, Hodgkin's, adult lymphoma, Hodgkin's, childhood lymphoma, He Jiejin Pregnancy, lymphoma, non-Hodgkin's, adult lymphoma, non-Hodgkin's, childhood lymphoma, non-Hodgkin's, lymphoma of pregnancy, primary central nervous system macroglobulinemia, Waldenstrom's malignant fibrous bone histiocytoma / osteosarcoma medulloblastoma, childhood melanoma melanoma 5 intraocular (eye) Merkel cell carcinoma mesothelioma, adult malignant Skin tumor, children with primary metastatic squamous cell carcinoma, multiple endocrine neoplasia syndrome group, children with multiple bone tumor / plasmacytoma verrucous granuloma bone marrow dysplasia syndrome bone marrow development / Myeloproliferative myelogenous leukemia, mattress #

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顱外胚細胞腫瘤，兒童骨髓性白血病，成人急性性腺外胚細胞腫瘤骨髓性白血病，兒童急性肝外膽管癌骨髓瘤，多發性眼癌，眼内黑素瘤骨髓增生病症，慢性眼癌，視網膜母細胞瘤鼻腔及副鼻竇癌膽囊癌鼻咽癌胃癌鼻咽癌，兒童胃癌，兒童神經母細胞瘤胃腸道類癌腫瘤非何杰金氏淋巴瘤，成人胚細胞腫瘤，顱外，兒童非何杰金氏淋巴瘤，兒童胚細胞腫瘤，性腺外非何杰金氏淋巴瘤，懷孕期胚細胞腫瘤，卵巢非小細胞肺癌姓娠滋養層腫瘤口癌，兒童神經膠瘤，成人口腔癌’唇及口咽癌神經膠瘤，兒童腦幹骨肉瘤/惡性纖維性骨組織細胞瘤神經膠瘤，兒童大腦即秦癌，兒童星狀細胞瘤卵巢上皮癌神經膠瘤，兒童視覺徑路及下視丘卵巢胚細胞腫瘤皮膚癌(黑素瘤）卵巢低惡性潛在腫瘤皮膚癌，默可細胞騰癌小細胞肺癌胰癌，兒童小腸癌胰癌，胰島細胞軟組織肉瘤，成人副鼻竇及鼻腔癌軟組織肉瘤，兒童副曱狀腺癌鱗狀細胞癌，參考皮膚癌(非黑素瘤）陰莖癌有隱藏原發性之鱗狀細胞頸癌，轉移胃癌胃癌，兒童嗜鉻細胞瘤松果體母細胞瘤及小腦幕未分化神經小腦幕上未分化神經外胚層腫瘤，外胚層腫瘤，兒童腦垂腺腫瘤 T細胞淋巴瘤，皮膚，參考蕈狀肉芽腫及謝薩瑞症候群漿細胞瘤/多發性骨髓瘤睪丸癌胸膜肺母細胞瘤懷孕及乳癌胸腺瘤，兒童胸腺瘤及胸腺癌懷孕及何杰金氏淋巴瘤甲狀腺癌甲狀腺癌，兒童懷孕及非何杰金氏淋巴瘤原發性中樞神經系統淋巴瘤攝護腺癌腎盂與輸尿管之過渡細胞癌滋養層腫瘤，妊娠直腸癌未知原發位置，癌，成人腎細胞(腎)癌腎細胞(腎)癌’兒童 73 200831525 未知原發位置，癌，兒童腎盂及輸尿管，過渡細胞癌兒童之非尋常癌視網膜母細胞瘤輸尿管及腎盂，過渡細胞癌橫紋肌肉瘤，兒童尿道癌唾液腺癌子宮癌，子宮内膜唾液腺癌，兒童子宮肉瘤肉瘤，歐文氏腫瘤家族 ***癌肉瘤，卡波西氏視覺控路及下視丘神經膠瘤，兒童肉瘤，軟組織，成人陰門癌肉瘤，軟組織，兒童沃登史壯氏巨球蛋白血症肉瘤，子宮威姆氏(Wilms’)腫瘤謝薩瑞症候群皮膚癌(非黑素瘤）皮膚癌’兒童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 5 經由抑制NFkB發揮作用；2-甲氧基***影響微管的形成及缺氧誘導因子(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麟醯胺、紫杉烧類 — (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. · 10 U.S.A” 1994e 91: ρ·3964-3968; D’Amato RJe等人，proe:Cranial blastoma, childhood myeloid leukemia, adult acute gonadal blast cell tumor myelogenous leukemia, childhood acute extrahepatic cholangiocarcinoma myeloma, multiple eye cancer, intraocular melanoma myeloproliferative disorder, chronic eye cancer, retina Bacteroma, nasal and paranasal sinus cancer, gallbladder cancer, nasopharyngeal carcinoma, gastric cancer, nasopharyngeal carcinoma, childhood gastric cancer, childhood neuroblastoma, gastrointestinal carcinoid tumor, non-Hodgkin's lymphoma, adult blast tumor, extracranial, child Jijin's lymphoma, childhood blastoma, extragonadal non-Hodgkin's lymphoma, gestational blast cell tumor, ovarian non-small cell lung cancer surname gestational trophoblastic cancer, childhood neuroglioma, adult oral cancer 'lip And oropharyngeal cancer neuroglioma, children's brain stem osteosarcoma / malignant fibrous bone histiocytoma neuroglioma, children's brain is Qin cancer, children's stellate cell tumor ovarian epithelial cancer neuroglioma, children's visual path and lower vision Mound ovary blast cell tumor skin cancer (melanoma) ovarian low malignant potential tumor skin cancer, silent cell cancer Lung cancer pancreatic cancer, childhood small intestine cancer pancreatic cancer, islet cell soft tissue sarcoma, adult paranasal sinus and nasal cancer soft tissue sarcoma, child parasitic adenocarcinoma squamous cell carcinoma, reference skin cancer (non-melanoma) penile cancer has hidden primary Squamous cell carcinoma of the squamous cell, metastasis of gastric cancer, pheochromocytoma of children, pediatric pheochromocytoma, and undifferentiated neuroectodermal tumors of the cerebellum, ectodermal tumor, squamous cell tumor of children Lymphoma, skin, reference verrucous granuloma and Xie Saray syndrome plasmacytoma / multiple myeloma testicular cancer pleural pulmonary blastoma pregnancy and breast cancer thymoma, childhood thymoma and thymic cancer pregnancy and Hodgkin's lymphoma Thyroid cancer thyroid cancer, childhood pregnancy and non-Hodgkin's lymphoma primary central nervous system lymphoma prostate cancer renal pelvis and ureteral transitional cell carcinoma trophoblastic tumor, pregnancy rectal cancer unknown primary location, cancer, adult kidney Cell (kidney) cancer, renal cell (kidney) cancer, 'Children 73 200831525 Unknown primary location, cancer, children's pelvis and urinary , transitional cell carcinoma, abnormal cancer, retinoblastoma, ureter and renal pelvis, transitional cell carcinoma, rhabdomyosarcoma, childhood urethral cancer, salivary gland cancer, uterine cancer, endometrial salivary gland cancer, childhood uterine sarcoma sarcoma, Owen's tumor family vaginal carcinosarcoma, Kaposi's visual control and hypothalamic neurofibroma, childhood sarcoma, soft tissue, adult genital sarcoma, soft tissue, children's Woden's giant globulin sarcoma, uterus Wilms' tumor Sari syndrome skin cancer (non-melanoma) Skin cancer 'Children In other embodiments of the invention, the compounds disclosed herein are useful for treating diseases associated with angiogenesis. Anti-angiogenic small molecules include thalidomide, part 5 of which acts through inhibition of NFkB; 2-methoxyestradiol affects microtubule formation and activation of hypoxia-inducible factor (HIFla); Enzyme 2 (COX2) inhibitors; and low-dose conventional chemotherapeutic agents include cyclamin, taxanes, and periwinkle bioassay (vincristine, vinbrastine) Vinblastine)) (D'Amato RJ· et al., Proc. Natl. Acad. Sci. · 10 USA” 1994e 91: ρ·3964-3968; D'Amato RJe et al., proe:

Natl· Acad. Sci· U.S.A·，1994. 91: p.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 15 (Glivec))及艾瑞莎(Iressa) (Bergers G.等人，J Clin Invest, 2003· 111: ρ·1287_1295; Ciardiello F.等人，臨床癌症研究， 74 200831525 臨床癌症研究，2003. 2001. 7: ρ·1459-1465; Plum S.M·等人， 9: p.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 5之治療方法。晚近，阿法斯丁 (八观⑷―種對血管内皮生長因子(VEGF)有高度親和力之抗體，阿法斯丁顯示用於亞化的腎細胞癌作為單-藥劑可延長壽命，用於惡化的大腸Natl·Acad. Sci·U.S.A., 1994. 91: p.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. These drugs include Herceptin, imatinib (Glivec) and Iressa (Bergers G. et al., J Clin Invest, 2003. 111: ρ · 1287_1295; Ciardiello F. et al., Clinical Cancer Research, 74 200831525 Clinical Cancer Research, 2003. 2001. 7: ρ·1459-1465; Plum SM· et al, 9: p. 4619-4626). Recently, angiogenesis inhibitors have been moved from animal research models to human patient studies. Angiogenesis inhibitors represent a variety of different cancers with a prospective 5 treatment. Lately, Affostine (Eight View (4) - a highly affinitive antibody to vascular endothelial growth factor (VEGF), Affintin shows that renal cell carcinoma for sub-induction as a single-agent can prolong life and be used for deterioration Large intestine

癌結合化學治療可延長壽命(YangJC等人，新英格蘭期刊Cancer combined with chemotherapy can prolong life (YangJC et al., New England Journal

Med, 2003. 349: p.427-434; Kabbinavar F.^ Λ，J Clin One 10 2003· 21: p.60-65) 〇 ’ 血管新生相關疾病包括但非限於發炎病、自體免疫病及傳染病；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、聽神經瘤、神經纖維瘤、氣管瘤、及產膿性肉芽腫；類風 15濕性關較；乾癖；濕療；目艮企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-偉博(〇sler_Webber)症候群；心肌血管新生；斑塊新生血管化；毛細血管擴張；嗜血性關節；血管纖維瘤；及傷 20 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、疣及肥厚性瘢痕（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 (Rochele minalia quintosa))、潰瘍（胃幽門螺旋桿菌 75 200831525 (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，包括癌症實施例及如此處揭示之曲西立濱化合物及/或一種或多 5 種鈾化合物。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 1〇也可能特有地發生於某些癌症而未先前暴露於化學治療劑。如此，於一個實施例中，本發明提供藉投予如此處揭不之TCN、TCN-P或相關化合物及一種或多種鉑化合物治療患有抗藥性癌症例如多重抗藥性癌症之病人之方法。於若干實施例中，TCN、TCN-P及相關化合物及一種或多種 15鉑化合物可用於治療單獨對紫杉醇、拉帕黴素、塔莫西芬' 西鉑汀及/或傑費堤尼（艾瑞莎）有抗藥性之癌症。於一個實施例中，TCN、Tc财或相關化合物及如此處揭不之-種或多種鈾化合物可用於治療抗藥性結腸癌、骨癌、腎癌、腎上腺癌、胰癌、肝癌及/或任已知或此處所述之癌症。技藝界 6·2·組合治療於一個實施例中，曲西立濱化合物及-種或多種鉬化合物可連同其它細胞毒劑投予。於另一個實施例中，曲西立m化合物及-種或多種麵化合物及其組成物當用於治療 76 200831525 實體腫瘤時，可使用輻射投予。於本1明之另-個實施例中，此處揭示之曲西立濱化口物及m或多種麵化合物及其組成物可組合至少一種額 ’ 耗學治療劑投？。額外_可與此錢示之化合物組合 : 5 &予或又替投予。樂物可構成同種組成物之-部分，或可於同時或不同時呈分開組成物投予。 … 於一個實施例中’如此處揭示之曲西立濱化合物及一種或多種翻化合物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 10另一個實施例中，曲西立濱化合物及一種或多種鉑化合物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（captopryi)、抗腫瘤劑諸如^干擾素、 ‘ 15 COMP (環磷醯胺、文克里斯丁、甲胺喋呤及普尼松 (prednisone))、伊妥普賽(et〇poside)、mBACOD (甲胺嗓呤、 _ 布利歐黴素(bleomycin)、朵索魯冰心（doxorubicin)、環填醯胺、文克里斯丁及德沙美沙松（dexamethasone》、 PRO-MACE/MOPP (普尼松、甲胺喋呤(含路可文(leuc〇vin) 20 救援）、朵索魯冰心、環磷醯胺、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、泰希闌（Techgalan)、沙利寶邁、 77 200831525 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絡元素，包括鬼臼毒或長春花生物鹼(文克里斯丁、文布拉斯丁）；抗代謝藥物（諸如 5 5-氟尿嘧啶、賽它拉冰（Cytarabine)、傑西塔賓 (gemcitabine)、嘌呤類似物諸如戊抑制素(pent〇statin)、甲胺喋呤）；烷化劑或氮芥子氣(諸如亞硝基脲、環構醯胺、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、 10 法母魯冰心(pharmombicin)或伊皮魯冰心(epirubicin);靶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（f]utamide)、路普瑞林 (leupimelin)、葛瑟瑞林(goserelin)、賽普左（cypr〇tr〇㈣或 15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（〇xaliplatin)、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· 20抗體治療；其中海洋來源之生物活性化合物值得注意^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(諸如乙酽胺酚(acetaminophen)或伊布普芬(ibupr〇fen))或類固醇及^ 何生物特別為德沙美沙松；止吐藥包括5HT_3抑制劑（諸如 78 200831525 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2所揭示之化學治療劑組合投予或交 ’ 替投予。表2 :化學治療劑 -13-順-維生素A酸 -2-胺基-6-疏基嗓呤 -2-CdA -2-氣去氧腺苔-5-氟尿嘴唆 -5-FU -6-TG -6-鳥嘌呤 -6-魏基嘌呤 -6-MP -阿查坦(Accutane) -放線徽素(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酸干擾素 -亞特塔明(Altretamine) 何米索泰林(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 _ -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) -乙酸歐萃泰 -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-天冬醯胺酶 -苯基丙二酸芥子氣 -普拉堤諾(Platinol)_ 79 200831525 -Ara-C -普拉堤諾-AQ -阿拉尼斯普(Aranesp) -普尼松隆(Prednisolone) -阿瑞戴(Aredia) -普尼松(Prednisone) -阿瑞米戴斯(Arimidex) -普隆(Prelone) -阿羅馬辛(Aromasin) -普卡巴辛(Procarbazine) -三氧化砷 -PROCRIT -天冬醯胺酶 -普路金(Proleukin) -ATRA -有卡幕斯丁(Carmustine)植體之普利菲 -阿法斯丁 (Avastin) 普斯潘(Prolifeprospan) 20 -BCG -普瑞尼索(Purinethol) -BCNU -拉羅西芬(Raloxifene) -貝法西組馬(Bevacizumab) _ 魯馬萃(Rheumatrex) -貝薩羅丁(Bexarotene) _瑞土山(Rituxan) •拜卡路塔邁(Bicalutamide) -瑞土西馬(Rituximab) •BiCNU -瑞福隆(Roveron)-A (干擾素a -2a) -布諾山(Blenoxane) -魯貝(Rubex) -布歐黴素(Bleomycin) -鹽酸魯畢朵黴素(Rubidomycin) -布特左米(Bortezomib) -山朵史塔丁 (Sandostatin) -布蘇坊(Busulfan) -山朵史塔丁 LAR -布蘇費(Busulfex) -薩葛幕斯丁 (Sargramostim) -C225 -蘇路-可堤(Solu-Cortef) -路可福瑞(Leucovorin)妈 -蘇路-美左(Solu-Medrof) -坎帕斯(Campath) -STI-571 -坎妥薩(Camptosar) -史翠妥左辛(Streptozocin) -喜樹驗(Camptothecin)-11 -塔莫西芬(Tamoxifen) -開普西塔冰(Capecitabine) 塔可丁 (Targretin) -卡拉(Carac) -紫杉醇(Taxol) -卡朋伯丁(Carboplatin) -紫杉帖(Taxotere) -卡幕斯丁 (Carmusiine) -堤莫達(Ternodar) -卡幕斯丁糊 -堤莫左羅米(Temozolomide) -卡索戴(Casodex) -堤尼普賽(Teniposide) -CCNU -TESPA -CDDP -沙利竇邁(Thalidomide) -西努(CeeNU) -薩羅米(Thalomid) -西魯比丁(Cerubidine) -席拉賽(TheraCys) -西土席馬(cetuximab) -硫鳥嘌呤 -可羅拉布席(Chlorambucil) -硫鳥嘌呤錠 -西 16 汀(Cisplatin) -硫石粦醯胺(Thiophosphoamide) -水化葉酸因子(Citrovorum Factor) -硫普歹1J(Thioplex)Med, 2003. 349: p.427-434; Kabbinavar F.^ Λ, J Clin One 10 2003· 21: p.60-65) 〇' Angiogenesis-related diseases include, but are not limited to, inflammatory diseases, autoimmune diseases and Infectious diseases; angiogenesis-dependent cancers include, for example, solid tumors, hematological tumors such as leukemia and tumor metastasis; benign tumors such as hemangioma, acoustic neuroma, neurofibromatosis, bronchoma, and pyogenic granuloma; Comparison; cognac; moist treatment; witnessing new diseases such as diabetic retinopathy, recurrent retinopathy, macular degeneration, corneal transplant rejection, neovascular glaucoma, posterior lens hyperplasia, iris redness; Oslo - Weibo (〇 sler_Webber) syndrome; myocardial angiogenesis; plaque neovascularization; telangiectasia; blood-sexual joints; angiofibroma; and 20 granulomas. In addition, 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., neoplasms). The composition of the present invention can also be used for the treatment of conditions in which angiogenesis occurs due to pathological consequences, such as cat scratching (Rochele minalia quintosa), ulcer (Helicobacter pylori 75 200831525 (Helobacter pylori)), Tuberculosis and lupus. 6.1 Treatment of Drug-Resistant Tumors or Cancer The present invention provides compounds useful for the treatment of drug-resistant cancers, including cancerous embodiments and the triclinide compounds and/or one or more uranium compounds as disclosed herein. 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 may also occur uniquely in certain cancers without prior exposure to chemotherapeutic agents. Thus, in one embodiment, the invention provides a method of treating a patient having a drug-resistant cancer, such as a multi-drug resistant cancer, by administering a TCN, TCN-P or related compound as disclosed herein and one or more platinum compounds. In several embodiments, TCN, TCN-P and related compounds and one or more platinum compounds can be used to treat paclitaxel, rapamycin, tamoxetine, xiplatin and/or gefitini (ai Resica) is a cancer resistant drug. In one embodiment, TCN, Tc or related compounds and one or more uranium compounds as disclosed herein can be used to treat drug resistant colon cancer, bone cancer, kidney cancer, adrenal cancer, pancreatic cancer, liver cancer, and/or any A cancer known or described herein. Technical Fields 6.2 Combination Therapy In one embodiment, the triclinide compound and one or more molybdenum compounds can be administered in conjunction with other cytotoxic agents. In another embodiment, the tricinem compound and the one or more facial compounds and compositions thereof, when used to treat 76 200831525 solid tumors, can be administered using radiation. In another embodiment of the present invention, the tromethonine reticulated substance and the m or a plurality of facial compounds and compositions thereof disclosed herein may be combined with at least one amount of therapeutic agent. . Additional _ can be combined with the compound of this money: 5 & The music may constitute a part of the same composition, or may be administered as separate components at the same time or at different times. In one embodiment, the triclinide compound and one or more compounds disclosed herein may be combined with an anti-angiogenic agent to enhance the effect, or may be administered in combination with other anti-angiogenic agents, along with other cytotoxic agents. In another embodiment, the triclinide compound and one or more platinum compounds and compositions, when used in the treatment of a solid tumor, can be administered with an agent selected from the group consisting of, but not limited to, IL-12, vitamin A, Interferon, angiostatin, endostatin, salipirin, thrombospondin-1, thromboxane-2, captopryi, antitumor agents such as interferon, '15 COMP (cyclophosphamide, vinsamine, methotrexate and prednisone), ettoposide, mBACOD (methylamine guanidine, _ bleomycin) , Doxorubicin, Doxorubicin, Cyclomethamine, Desmetthasone, PRO-MACE/MOPP (Penison, Methotrex (including Leuc〇vin) ) 20 Rescue), Dosso Lu Bing Xin, Cyclophosphamide, Iptosemine, Mechlorethamine, Venlistin, Pnisson and Procarbazine, Wen Christine , vebrabutin, angiostatin, TNP-470, pentosan polysulfate, platelet factor 4, angiostatin, LM- 609, SU-101, CM-101, Techgalan, Shalibaomai, 77 200831525 SP-PG and radiation. In additional embodiments, the compounds and compositions disclosed herein can be combined with the following drugs Pre- or alternating administration: for example, drugs with anti-mitotic effects such as standard cell bone elements, including podophyllotoxin or vinca alkaloids (Wen Christian, von Brastin); antimetabolites (such as 5 5 -Fluorouracil, Cytarabine, gemcitabine, purine analogs such as pent〇statin, methotrexate; alkylating agents or nitrogen mustard gas (such as nitrosourea, Cyclohexylamine or ifosphamide; drugs targeting DNA such as antracycline drug adriamycin, dosoluru, and 10 pharmombicin Or epirubicin; drugs targeted at topoisomerases such as etatop; hormones and hormone agonists or hormone antagonists such as estrogens, antiestrogens (tamoxifen and related) Compound) and androgen, Fotamai (f]utam Ide), leupimelin, goserelin, cyp〇tr〇 (four) or 15 octreotide; drugs that target signal transduction of tumor cells, Including antibody derivatives such as carbamazepine; alkylating drugs such as platinum drugs (xiplatin, carbonplatin, 〇xaliplatin, paraplatin) or nitroso Ureas; drugs that may affect tumor metastasis, such as matrix metalloproteinase inhibitors; gene therapy and anti-reagents 20 antibody therapy; where marine-derived bioactive compounds are worth noting ^ are Didemnins such as Apridine ( Aplidine); a steroid analog, particularly dexamethasone; an anti-inflammatory drug including a non-steroidal agent (such as acetaminophen or ibuprofen) or a steroid and a bio-specific Desha Methadone; antiemetics include 5HT_3 inhibitors (such as 78 200831525 gramisetron or ondasetron) and steroids and their derivatives, especially dexamethasone. In still other embodiments, the compounds and compositions can be administered or administered in combination with the chemotherapeutic agents disclosed in Table 2 below. Table 2: Chemotherapeutic agent-13-cis-vitamin A-2-amino-6-mercaptopurine-2-CdA-2-gas deoxygenated gland-5-fluoropurine 唆-5-FU - 6-TG -6-guanine-6-weiki嘌呤-6-MP-Accutane-Acctinomycin-D•Adriamycin-Adrucil -Agrylin - Ala-Cort - Aldesleukin - Alemtuzumab - Alitretinoin - Alkaban - AQ - Alkeran - All-trans retinoic acid interferon - Altretamine Amethopterin - Amifostine _ Aminoglutethimide - Anagrelide Anandron - Anastrozole - Arabosidinoside - Neosar - Neulasta - Neumega - Nupkin ( Neupogen) - Nilandron • Nilutamide - Nitrogen mustard - Novaldex - Novantrone - Octreotide - Acetate - Oukes Acetate Oncospar _ Oncovin - Ontak - Onxal - Opvi Oprevelkin • Orapred - Orasone - Oxaliplatin - Paclitaxel - Pamidronate - Panretin - Parab Paraplatin - Pediapred - PEG Interferon - Pegaspargase • Pegfilgrastim - PEG-INTRON - PEG-L-aspartate-Benzene Malonic acid mustard gas - Platinol _ 79 200831525 -Ara-C -Platino-AQ -Aranesp -Prednisolone -Aredia - Prednisone - Arimidex - Prelone - Aromasin - Procarbazine - Arsenic Trioxide - PROCRIT - Aspartate - Pulu Proleukin -ATRA - Avastin with Carmustine implants Prolifeprospan 20 -BCG -Purinethol -BCNU - Pull Raloxifene - Bevacizumab _ Rheumatrex - Bexarotene _ Rituxan • Bicalutamide - Swiss Rituximab • BiCNU - Roveron-A (interferon a -2a) - Blenoxane - Rubex - Bleomycin - Rubicillin Hydrochloride Rubidomycin - Bortezomib - Sandostatin - Busulfan - Mountain Stalling LAR - Busulfex - Sargramostim ) -C225 - Solu-Cortef - Leucovorin Ma - Sulu - Solu - Medrof - Campas - STI-571 - Cantosa Camptosar) - Streptozocin - Camptothecin-11 - Tamoxifen - Capecitabine Targretin - Carac - paclitaxel Taxol) - Carboplatin - Taxotere - Carmusiine - Ternodar - Temozolomide - Caso Dai (Casodex) - Teniposide - CCNU -TESPA -CDDP - Thalidomide - CeeNU - Thalomid - Cerubidine - Syrah TheraCys - Cetuximab - Sulfur嘌呤 -Chlorambucil - Thioguanine ingot - Cisplatin - Thiophosphoamide - Citrovorum Factor - Thioplex

80 20083152580 200831525

-可拉左冰(Cladribine) -可體松(Cortisone) -可美金(Cosmegen) -CPT-11 -環磷醯胺(Cyclophosphamide) -賽塔左恩(Cytadren) -賽塔拉賓(Cytarabine) -賽塔拉賓微脂粒 -賽妥色(Cytosar)-U -賽妥山(Cytoxan) -達卡巴辛(Daearbazine) -達堤諾黴素(Dactinomycin) -達本普丁 (Darbepoetin) α -道諸黴素(Daunomycin) -鹽酸道謹魯冰心(paunorubicin) -道諾魯冰心微脂粒 -道諾索(DaunoXome) -德卡左(Decadron) -戴它可泰(Delta-Cortef) -戴它松(Deltasone) -戴尼魯金(Denileukin)戴堤妥(diflitox) -德普赛(DepoCyt) -德沙美沙松(Dexamethasone) -乙酸德沙美沙松 -磷酸德沙美沙松鈉 -德沙松(Dexasone) -戴拉左山(Dexrazoxane)- Cladribine - Cortisone - Cosmegen - CPT-11 - Cyclophosphamide - Cytadren - Cytarabine -赛塔拉宾脂脂脂-Cytosar-U-Cytoxan-Daearbazine-Dactinomycin-Darbepoetin α-Channel Daunomycin - Paunorubicin - Donovan Ice Core - DaunoXome - Decadron - Delta-Cortef - Wear it Deltasone - Denilukin diflitox - DepoCyt - Dexamethasone - Desamethasone acetate - Desamethasone sodium - Dessert (Dexasone) - Dexrazoxane

-DHAD-DHAD

-DIC -戴德 piodex) •朵西紫杉醇(Docetaxel) -朵席(Doxil) -朵索魯冰心(Doxorubicin) -朵索魯冰心微脂粒 -左赛亞(Droxia)-DIC-dade piodex) • Docetaxel - Doxil - Doxorubicin - Dorothy Ice Core - Droxia

-DTIC -DTIC-朵母(Dome) _ 杜拉隆(Duralone) -伊福戴(EfUdex)_ -硫泰帕(Thiotepa)-DTIC -DTIC-Dome _ Duralone - EfUdex _ - Thiotepa

-TICE -妥普薩(Toposar) -妥普堤肯(Topotecan) -妥瑞米芬(Toremifene) -崔茲竹美(Trastuzumab) -崔堤諾(Tretinoin) -崔薩(Trexall) -崔森諾(Trisenox)-TICE - Toposar - Topotecan - Toremifene - Trastuzumab - Tretinoin - Trexall - Cui Senno (Trisenox)

-TSPA-TSPA

-VCR -維班(Velban) -維卡德(Velcade) -維普西(VePesid) -維莎諾(Vesanoid) 維杜爾(Viadur) -硫酸文布拉斯丁 (Vinblastine) -文卡薩(Vincasar) Pfs -文克里斯丁 (Vincristine) -文諾瑞賓(Vinorelbine) -酒石酸文諾瑞賓-VCR -Velban -Velcade -VePesid -Vesanoid Viadur -Vinblastine - Vincasar Pfs - Vincentine - Vinorelbine - Venoribine

-VLB -VP-16 •福蒙(Vumon) -贊羅達(Xeloda) -山諾薩(Zanosar) -山法林(Zevalin) -辛卡(Zinecard) -左拉戴(Zoladex) -左朵尼酸(Zoledronic acid) 左米塔(Zometa) -葛里戴(Gliadel)糊 -葛里費(Glivec)-VLB - VP-16 • Vumon - Xeloda - Zanosar - Zevalin - Zinecard - Zoladex - Zuo Duoni Zoledronic acid Zometa - Gliadel paste - Glivec

-GM-CSF •葛瑟瑞林(Goserelin) •粒狀細胞-群落刺激因子 -粒狀細胞-巨嗟細胞-群落刺激因子 -哈羅泰斯丁 (Halotestin) -贺癌平(Herceptin) 81 200831525 -伊萊嘉(E%ard) -艾倫斯(^Ellence) -艾羅薩丁(Eloxatin) -艾斯巴(Elspar) -安賽特(Emcyt) -伊皮魯冰心(Epirubicin) -伊普丁 (Epoetin) α -爾畢妥(Erbitux) -艾文尼亞(Erwinia)L-天冬醯胺酶 -伊塔幕斯丁 (Estramustine) -伊賽歐(Ethyol) -伊妥普福(Etopophos) -伊妥普賽(Etoposide) -磷酸伊妥普赛 _優列辛(Eulexin) -艾文史塔(Evista) -伊淫米史坦(Exemestane) -法列史東(Fareston) -法史羅戴(Faslodex) -費馬拉(Femara) -費葛拉斯堤(Filgrastim) -福羅蘇瑞丁 (Floxuridine) -福達拉(Fludara) -福達拉賓(Fludarabine) -福羅普列(Fluoroplex) -氟尿嘧咬 -氟尿嘧啶(乳膏） -福賽米史特隆(Fluoxymesterone) -福塔邁(Flutarnide) -酸葉酸(Folinic Acid) -FUDR -福維斯特拉(Fulvestrant) -G-CSF -傑費堤尼(Gefitinib) -傑西塔賓(Gemcitabine) -傑土竹美(Gemtuzumab)歐索嘉米辛 (ozogamicin) -傑薩(Gemzar) -葛歹1J 費(Gleevec)_ -贺薩左(Hexadrol) -賀薩蘭(Hexalen) ••六曱基蜜胺 -HMM -海坎丁 (Hycamtin) -海左(Hydrea) -乙酸經可體(Hydrocort) -經可體松(Hydrocortisone) -填酸經可體松納 -丁二酸羥可體松鈉 -填酸經可酮(Hydrocortone) -經基脲(Hydroxyurea) -伊布土莫馬(Ibritumomab) -土西坦(Tiuxetan) -伊達黴素(Idamycin) -伊達魯冰心(Idarubicin) -伊福(Ifex) -IFN-a -伊福法邁(Ifosfamide) -IL-2 -IL-11 甲磺酸伊馬堤尼(Imatinib) -咪唑羧醯胺 -干擾素α -干擾素a-2b (PEG軛合物） -介白素-2 -介白素-11 -因壯(Intron) A (干擾素 a -2b) -魯可福林(Leucovorin) _魯克蘭(Leukeran) _ 魯凱恩(Leukine) -魯普萊(Leuprolide) -魯羅克里斯丁 (Leurocristine) -魯史塔丁 (Leustatin) -微脂粒Ara-C -液體普來德(Pred) -羅幕斯丁 (Lomustine) -L-PAM -L-莎可萊辛(Sarcolysin)_ 82 200831525 -魯普(Lupron) -米堤可坦(Meticorten) -長效戴普(Depot) -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 •美拉隆(Medralone) -MTX -美左(Medrol) ••幕斯塔金(Mustargen) -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) -巯基嘌呤 -伊瑞諾堤坎(Irinotecan) -美納(Mesna) -伊索特堤諾(Isotretinoin) -美淫(Mesnex) •吉左拉斯(Kidrolase) -曱胺嗓吟(Methotrexate) _拉那可(Lanacort) -曱胺嗓吟納 -L-天冬醯胺酶 •甲基普尼松隆(Methylprednisolone) -LCR -邁羅賽(Mylocel) -利妥左(Letrozole)-GM-CSF •Goserelin •Rye cell-community stimulating factor-granulocyte-maize cell-community stimulating factor-Halotestin-Herceptin 81 200831525 - Eyard - Ellence - Eloxatin - Elspar - Emcyt - Epirubic - Ip Epoetin α Erbitux - Erwinia L-aspartate - Estramustine - Ethyol - Etopophos -Etoposide - Elutinose Eulexin - Evista - Exemestane - Fareston - Fastron (Faslodex) - Femara - Filgrastim - Floxuridine - Fludara - Fludarabine - Fluoroplex - Fluorouracil-Fluorouracil (Cream) - Fluoxymesterone - Flutarnide - Folinic Acid - FUDR - Fulvestrant -G-CSF - Gefitinib - Jemitabin (Gemcitabi Ne) - Gemtuzumab ozogamicin - Gemzar - Gee 1J Fee (Gleevec) _ - Hexadrol - Hexalen • Six Mercapto melamine-HMM-Hycamtin-Hydrea-Hydrocort-Hydrocortisone-acid-filled cortisone-succinic acid hydroxyconductor Hydrocorretone - Hydroxyurea - Ibritumomab - Tiuxetan - Idamycin - Idarubicin - Ipho (Ifex) - IFN-a - Ifosfamide - IL-2 -IL-11 Imatinib mesylate - Imidazole carboxamide - interferon alpha - interferon a-2b (PEG yoke Compound) - Interleukin-2 - Interleukin-11 - Intron A (Interferon a -2b) - Leucovorin _ Leukeran _ Leukine ) - Leuprolide - Lerocristine - Leustatin - Lipid Ara-C - Liquid Pred - Lomustine - L -PAM -L-Sarcolysin_ 82 200831525 -Lupron -Meticorten - Depot - Mitomycin - Matulane - mirtamycin - C - Maxidex - Mitoxantrone - Mercury Samin ( Mechlorethamine) - Prednisol - Mecorosamin Hydrochloride - MTC • Medralone - MTX - Medrol • Mustargen - Megace _ Mustine - Megestrol - Mutamycin - Myleran _ Melphalan - Iressa - 巯基嘌呤-Irinotecan - Mesna - Isotretinoin - Mesnex • Kidrolase - Methotrexate _ Lanaco (Lanacort) - Alanine Cannes - L - Aspartate - Methylprednisolone - LCR - Mylocel - 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a -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素召、干擾素r、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（Human Gen〇me Sciences)製造、瑞畢福(REBIF)(干擾素0-ia)艾瑞斯色隆 10諾公司（Ares-Seirono)製造、ω干擾素生醫公司（BioMedicine) 製造、口服干擾素（2亞馬雷洛生科公司（Amarillo Biosciences)製造及干擾素7、干擾素r、及/或干擾素 83 200831525 点因特幕公司製造。於一個實施例中，如此處揭示之TCN、TCN-P或相關化合物及一種或多種鉑化合物可與額外化學治療劑諸如此處所述或表2所示之額外化學治療劑組合使用或交替使用 5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、月癌、腎癌、腎上腺癌、騰癌、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 10 (verapamil)、賽羅史寶靈（CyCi〇Sp〇rin)(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（dexverapamil)、戴尼谷戴平（dexniguldipine)、凡史普達 (valspodar) (PSC 833)、拜瑞可達(biricodar) (VX-710)、塔瑞奎達（tariquidar) (XR9576)、左蘇奎達（zosuquidar) 15 (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一種或多種鉑化合物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 20 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。曲西立續化合物且與一種或多種翻化合物組合調配成為組成物中之唯一藥學活性成分或可與一種或多種鉑化合物組合調配。 84 200831525 包合物及—種或多賴化合物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、腦池內、胳咖 ^ _ 自也内、腹内、及硬膜外)投藥。較佳組 5 成物係經靜脈投藥。In several embodiments, interferon (IFN) can be used in combination with a compound of the invention. Suitable interferons include: interferon a-2a, interferon a-2b, pegylated interferon alpha including interferon a-2a and interferon alpha _2b, interferon/3, interferon 5, interferon r, interferon ω, INFERGEN (interferon a con-1) manufactured by InterMune, manufactured by OMNIFERON (natural interferon) Veragen, Ou Buffon (ALBUFERON) manufactured by Human Genology Scientific Corporation, REBIF (Interferon 0-ia) manufactured by Ares-Seirono, Omega Interferon Biomedical Manufactured by the company (BioMedicine), oral interferon (2 manufactured by Amarillo Biosciences and interferon 7, interferon r, and/or interferon 83 200831525 manufactured by Deutschland Co., Ltd.) The TCN, TCN-P or related compound and one or more platinum compounds as disclosed herein may be used in combination with an additional chemotherapeutic agent such as the additional chemotherapeutic agents described herein or in Table 2, or used interchangeably to treat the antibody. Drug-induced cancers such as multi-drug resistant cancers. Drug-resistant cancers include large Cancer, monthly cancer, kidney cancer, adrenal cancer, 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-glycoprotein inhibitor. In several non-limiting embodiments, the 'P-glycoprotein inhibitor can be selected from the group consisting of verapamil, CyCi〇Sp〇rin (such as Cyrus Blingling A), and the tower. Moxifene, calmodulin antagonist, dexverapamil, dexniguldipine, valspodar (PSC 833), biricodar (biricodar) VX-710), tariquidar (XR9576), zosuquidar 15 (LY335979), laniqUidar (R101933) and/or ONT-093. A composition comprising a trichostatin compound and one or more platinum compounds can be administered, if desired, with a pharmaceutically acceptable carrier or excipient. Suitable for administration of a pharmaceutically acceptable carrier, such as a compound provided herein. Included by those skilled in the art who are known to be suitable for a particular mode of administration Such a carrier, a compound of trichostatin and formulated in combination with one or more compounds, is the sole pharmaceutically active ingredient in the composition or may be formulated in combination with one or more platinum compounds. 84 200831525 Inclusion compounds and compositions of the compounds or compounds of the compounds are suitable for oral, rectal, nasal, transdermal (including buccal and sublingual), transvaginal or parenteral (including subcutaneous, intramuscular, Subcutaneous, intravenous, intradermal, intraocular, intrabronchial, intracranial, gynecological _ _ self, intra-abdominal, and epidural. Preferably, the group 5 is administered intravenously.

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-種或多種本發明組成物與-種或多種藥學載劑或賦形劑址合之步驟。曲西立濱化口物及—種或多種翻化合物及其組成物可 10調配成適當藥學製劑，諸如溶液劑、懸浮液劑、錠劑、分散鍵劑:丸劑、膠囊劑、散劑、持續釋放配方、或触劑供、、’工口投藥，或可壬無_溶液劑或懸浮液劑供腸道外投藥；以及經皮貼片製劑及乾粉吸入劑。於一個實施例中，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 is配成藥學組成物(例如參考Ansd，#學劑型導論，第*版， 1985 年，126頁）。於該組成物中，有效濃度之-種或多種化合物或其藥學上可接受之衍生物可與一種或多種適當藥學載劑混合。本發明化合物可於調配前衍生成相對應之鹽類、酯類、烯 2〇醇醚類或烯醇酯類、縮醛類、縮酮類、原酸酯類、半縮醛類、半縮酮類、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一種或多種症狀之用 ϊ。於一個實施例中，組成物調配供單劑投藥。為了調配 85 200831525 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，讓所治療的病情被缓解、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 5錠劑、橢圓錠、丸劑、或糖衣劑、膠囊劑、或扁囊劑，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。液體藥學可投予組成物之製法例如可經由將曲西立濱 10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 4，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辅助性物質諸如濕潤劑、乳化劑、增溶劑、pH緩衝劑、保藏劑、矯味劑等，例如乙酸鹽、檸 15檬酸鈉、環糊精衍生物、一月桂酸山梨聚糖酯、三乙醇胺乙酸納、二乙醇胺油酸酯、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。適合局部投予口部之本發明組成物例如包括***錠， 20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一種或多種鉑化合物於惰性基劑諸如明膠及甘油或蔗糠及金合歡膠；及漱口藥，其含有一種或多種本發明之組成物於適當液體載劑投予。 86 200831525 錠劑、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 - 5 膠、葡萄糖溶液、金合歡黏質、明膠溶液、糖蜜、聚乙烯基比嘻唆酮、普維隆(povidone)、交聯普維隆、蔗糖及殿粉二糊。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、 Φ 澱粉、高嶺土、鹽、甘露糖醇、及磷酸二鈣。滑動劑包括 10 但非限於膠體二氧化矽。崩散劑包括交聯曱基纖維素鈉、乙醇酸澱粉鈉、褐藻酸、玉米澱粉、馬鈐薯澱粉、膨潤土、曱基纖維素、瓊脂、及羧甲基纖維素。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；及懸浮於水合礬土上之水不溶性FD&C染料。甜味劑包括蔗 " 15 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 β 劑，其可產生怡人感覺之化合物之合成掺合物，諸如但非限於薄荷腦及水揚酸甲酯。濕潤劑包括一硬脂酸丙二醇酯、一油酸山梨糖醇酯、一月桂酸二乙二醇酯及聚氧伸乙 20 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、及乙酸磷苯二甲酸纖維素。膜衣包括羥乙基纖維素、羥曱基纖維素鈉、聚乙二醇4000及乙酸磷苯二甲酸纖維素。適合局部投予皮膚之組成物可呈有一種或多種組成物 87 200831525 於樂學上可接受之載劑投予之軟膏劑、乳膏劑辞膠劑及糊劑劑型。直腸投藥用組成物可呈含適當基可可脂或水揚酸酯之栓劑劑型投予 5 適合供經鼻投藥之組成物，當載劑為固體時，、、，組成物已括具有20微米至500微米範圍之粒徑之粗粉，以〇 + 1 鼻嗅粉之方式投藥（亦即將粉末容器保持接近鼻孔快迷、、通過 π道投藥）。當載劑為液體(例如鼻噴霧劑或鼻滴劑）時，一種或多種組成物可混合於水性溶液或油性溶液而煲入畠省 10 或噴霧進入鼻道。冗、適合供***投藥之組成物可呈含有一種戋吝於、 7赞組成物及L Μ載劑之子宮托、棉塞、軟膏劑、明膠劑、糊劑、發泡劑或噴霧配方劑型。適合供腸道外投藥之組成物包括水性及非水性無菌、主 15射溶液劑，其可含有抗氧化劑、緩衝劑、制…、囷/主 ^ ΡΗι 方S:成與期望接受者之血液呈等張性之溶質，及水丨生無— 懸浮液劑及非水性無菌懸浮液劑，其包括懸浮劊劑。組成物可呈單劑或多劑容器例如密封安瓿及冒周形式，組成物可儲存於冷凍乾燥（凍乾)條件下，只需恰在使 20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子液劑可由前文說明之該種無菌散劑、粒劑及旋劑製備〇 . 適合供腸道投藥或腸道外投藥之藥用有機或無機固體或液體載劑介質可用來製造組成物。明膠、乳糠、殿粉、 200831525 硬脂_、滑石、植物及動_肪類及油類、樹膠、 :基一醇、水、或其它已知载劑全部皆適合用作為載劑介 5The compositions may conveniently be presented in unit dosage form or may be prepared by conventional pharmaceutical techniques. Such techniques include the step of bringing into association one or more of the compositions of the present invention with one or more pharmaceutical carriers or excipients. Quci Libin sulphate and one or more compounds and compositions thereof can be formulated into appropriate pharmaceutical preparations, such as solutions, suspensions, lozenges, dispersing agents: pills, capsules, powders, sustained release Formulation, or contact agent supply, 'work administration', or 壬 solution or suspension for parenteral administration; and transdermal patch preparation and dry powder inhaler. 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, see Ansd, Introduction to Academic Formulations, ed., 1985, p. 126). In the composition, an effective concentration of the compound or compounds or a pharmaceutically acceptable derivative thereof may be combined with one or more suitable pharmaceutical carriers. The compound of the present invention can be derivatized into corresponding salts, esters, olefinic sterol ethers or enol esters, acetals, ketals, orthoesters, hemiacetals, hemi-condenses before preparation. Ketones, acids, bases, solvates, hydrates, or prodrugs. The concentration of the compound in the composition can be effectively delivered at the time of administration to treat or prevent one or more symptoms of the target disease or condition. In one embodiment, the composition is formulated for administration in a single dose. To formulate the composition of 2008 200831525, the weight component of the compound is dissolved, suspended, dispersed or otherwise mixed at a concentration effective to the selected carrier to delay, prevent, or ameliorate one or more symptoms of the condition being treated. Suitable oral pharmaceutical compositions may be in separate units such as, but not limited to, 5 tablets, elliptical tablets, pills, or dragees, capsules, or cachets, each containing a predetermined amount of one or more of the compositions; Or a granule; a solution or suspension in 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. Liquid pharmaceutical pharmaceutically acceptable compositions can be prepared, for example, by dissolving, dispersing or otherwise mixing the triclinide 10 compound and optional pharmaceutical adjuvants in carriers such as water, saline, aqueous dextrose, glycerol, glycol Class, ethanol 4, 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, solubilizers, pH buffering agents, preservatives, flavoring agents, etc., such as acetate, lemon 15 sodium citrate , cyclodextrin derivatives, sorbitan monolaurate, sodium triethanolamine acetate, diethanolamine oleate, and other such adjuvants. Such dosage forms are known to those skilled in the art or are readily apparent, for example, by reference to Remington's Pharmaceutical Sciences, Merck Publishing Company, Easton, Pennsylvania. Compositions of the invention suitable for topical administration to the mouth include, for example, buccal tablets, 20 which contain the ingredients in a flavoring base, typically sucrose and acacia or tragacanth; soft tablets, which contain one or more of the present invention a sirolimus compound and one or more platinum compounds in an inert base such as gelatin and glycerin or cane and acacia; and a mouthwash containing one or more of the compositions of the invention administered in a suitable liquid carrier . 86 200831525 Tablets, pills, capsules, throat tablets, etc. may contain one or more of the following ingredients or compounds of similar nature: binders; lubricants; diluents; slip agents; disintegrating agents; colorants; sweeteners; Agent; humectant; reminder 'spray coating; and film coat. Examples of the binder include microcrystalline cellulose, scutellaria gel-5, glucose solution, acacia viscous, gelatin solution, molasses, polyvinylpyrrolidone, povidone, crosslinked Pvillon , sucrose and two powder paste. Lubricants include talc, starch, magnesium stearate or calcium stearate, lycopodium, and stearic acid. The diluent includes, for example, lactose, sucrose, Φ starch, kaolin, salt, mannitol, and dicalcium phosphate. The slip agent includes 10 but is not limited to colloidal cerium oxide. Disintegrating agents include crosslinked bismuth cellulose sodium, sodium starch glycolate, alginic acid, corn starch, horse starch starch, bentonite, sulfhydryl cellulose, agar, and carboxymethyl cellulose. The coloring agent includes, for example, any of the approved and approved water-soluble FD&C dyes, a mixture thereof; and a water-insoluble FD&C dye suspended on hydrated alumina. Sweeteners include any of sugar cane <15 sugar, lactose, mannitol, and artificial sweeteners such as saccharin and spray dry flavors. Flavoring agents include flavoring agents derived from plants such as fruits which produce a pleasant blend of compounds such as, but not limited to, menthol and methyl salicylate. The humectant includes propylene glycol stearate, sorbitan monooleate, diethylene glycol monolaurate, and polyoxyethylene 20-based lauryl ether. The emetic coating includes fatty acids, fats, waxes, shellac, ammoniated shellac, and cellulose acetate phthalate. The film coat includes hydroxyethyl cellulose, sodium hydroxymethyl cellulose, polyethylene glycol 4000, and cellulose acetate phthalate. Compositions suitable for topical administration to the skin may be in one or more compositions. 87 200831525 Essences, creams, gelatins and paste formulations for administration of a carrier which is acceptable for learning. The rectal pharmaceutical composition may be administered as a suppository formulation containing a suitable base of cocoa butter or salicylate. 5 A composition suitable for nasal administration. When the carrier is a solid, the composition has been included to have a thickness of 20 μm. The coarse powder of the particle size of 500 micrometers is administered as a sputum + 1 nasal olfactory powder (ie, the powder container is kept close to the nostrils, and is administered by the pi tract). When the carrier is a liquid (e.g., a nasal spray or a nasal drop), one or more of the compositions can be mixed with an aqueous or oily solution to invade or spray into the nasal passages. A composition which is cumbersome and suitable for vaginal administration may be in the form of a pessary, tampons, ointment, gelatin, paste, foaming or spray formulation containing a sputum, a scented composition and an L sputum carrier. Compositions suitable for parenteral administration include aqueous and non-aqueous sterile, primary 15 solutions, which may contain antioxidants, buffers, preparations, 囷/main^ ΡΗι S: into the blood of the intended recipient, etc. The solute of the puberty, and the sputum of the sputum - suspensions and non-aqueous sterile suspensions, including suspension elixirs. The composition may be in the form of a single or multiple dose container such as a sealed ampoule and a weekly form, and the composition may be stored under lyophilization (lyophilization) conditions, just prior to the use of a sterile liquid carrier such as water for injection. The temporary injection solution and the suspension solution can be prepared from the above-mentioned sterile powder, granules and granules. The medicinal organic or inorganic solid or liquid carrier medium suitable for enteral administration or parenteral administration can be used for the manufacture of the composition. Things. Gelatin, milk thistle, temple powder, 200831525 Hard fat _, talc, plant and animal fats and oils, gums, base alcohol, water, or other known carriers are suitable for use as a carrier.

10 包括曲西立濱化合物及一種或多懸化合物可組合—種或多種藥學上可接受之制介質及/錢= 使用。如此處使用，「藥學上可接受之制介f」包括^ 種及全部制、溶劑、稀釋劑、或以其它液體載媒劑、八散助劑或懸浮助劑、表面活性劑、等張劑、增稠劑刀劑、保藏劑、固雜結劑、騎劑、輔劑、載媒劑、糸統、崩㈣、吸㈣、保藏劑、界面活性劑、著、 =味劑、或甜味解皆依其適合用於期望之料劑型^ 15 20 =曲西立濱化合物及-種或多種翻化合物之可與f學上可接受之賦形劑及任選地持續釋放基 :如二生物分解聚合物組合來形成治療性組成物。「藥二上之::劑」包括無毒固體、半固體或液體過濾劑、片、囊封材料或任何類型之配方辅劑。醫療組成物之總每曰使用係由臨床醫師依據完整 :療判任何特定教之特定治療有效 ::::多項因素決定，例如包括所治療之病症及該病症 & ί又’所使用之特定組成物之活性；所使用之特定纪病人年齡、體重、-般健康狀況、性別及飲食；投樂《 ’投_徑；所使用之特定化合物之***速率；治〃所彳*用之特定組成物組合使用朗時使用之曲 200831525 西立濱化合物及/或一種或多種鉑化合物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低的劑量開始投予該組成物，以及徐緩提高劑量至達到期望功效。 5 包括曲西立濱化合物及一種或多種鉑化合物之組成物為了谷易投藥及劑量的均一，較佳係調配成單位劑型。如此處使用之「單位劑型」係指適合用於欲治療之宿主之組成物之貫體上分開單位。各劑量須含有計算來就此、或於所選藥學上載劑介質結合來產生期望之療效之組成物數 10 量。較佳單位劑量配方為含有所投予成分之每日劑量或每曰劑量單位、每曰次劑量或其適當分量之單位劑量配方。例如母日約1_5宅克此處揭示之化合物可縮小小鼠體内之實體腫瘤體積。 15 劑里將依據侣主因素諸如體重、年齡、表面積、代謝、組織分布、吸收速率及***速率決定。於一個實施例中，每曰約0.5克至7克如此處揭示之曲西立濱化合物可投予人類。任選地，母日約1克至4克曲西立濱化合物可投予人類。於若干實施例中，每曰約0.001毫克至5毫克曲西立濱化合 20物可投予人類。治療有效劑量將依據前文說明之多項因素決定。此外，於技藝界技巧範圍内可於相對低劑量開始投予組成物，以及提高劑量至達到期望的效果。包括曲西立濱化合物及一種或多種鉑化合物之組成物可用於持續釋放基體，基體可由可藉酵素水解或基於酸之 90 200831525 水解或藉溶解分解的材料製成，通常為聚合物。—旦*** 身體内部，基體受_及體液的作用。持續釋放基體例如係選自於可生物相容之材料諸如微練類、聚丙交酷類（聚乳酸）、聚乙交醋（乙醇酸聚合物）、聚兩交醋共聚合乙交醋 5 (乳酸與乙醇酸之共聚物）、聚酐類、聚(原酸)醋類、多耿類曰、玻尿酸、膠原蛋白、硫酸軟骨素、緩酸類、脂肪酸類、填脂類、多醣類、核酸類、多胺基酸類、胺基酸類諸如苯基丙胺酸、酪胺酸、異白胺酸m酸類、聚乙稀基丙婦土、聚乙稀基t各咬酮、及聚石夕氧。較佳可生物分解基體為聚 10丙交自旨、聚乙錢、絲丙交s旨共聚合乙錢（乳酸與乙醇酸之共聚物）中之一者之基體。曲西立濱化合物及-種或多種麵化合物也可呈微脂粒形式投予。如技藝界已知，微脂粒通常係衍生自碟脂或其它脂質物質。微脂粒係由單層或多層水合液晶分散於水性 15介質所形成。可使用可形成微脂粒之任—種無毒性生理上可接受性且可代謝之脂質。除了本發明之_種或多種組成物外，微脂粒可含有安定劑、保藏劑、賦形劑等。脂質之實例為天然及合成之磷脂類及_月旨基贍驗類（印彻旨類）。微脂粒之形成方法為技藝界眾所周知。 2〇自西立濱化合物及-種或多種鈾化合物可調配成氣溶膠供例如藉吸入方式施用。投予呼吸道之此等配方可呈氣溶膠或喷霧劑溶液形式或呈微細吹入用粉末形式，可單獨使用或組合惰性載劑諸如乳糠使用。於此種情況下，於一個實施例中，配方顆粒具有小於50微米之直徑，於一個實 91 200831525 施例中小於1 〇微米。包括曲西立濱化合物及一種或多種鈾化合物之組成物可與其它治療前述病情之組成物及/或程序組合使用。舉例言之，腫瘤習知係使用手術、放射性治療或化學治療組合 5 一種或多種本發明組成物處理，隨後一種或多種本發明組成物可投予該病人來延長微小腫瘤轉移的休眠，且穩定、抑制或減少任何殘留原發性腫瘤的生長。 7.1.额外實施例包括曲西立濱化合物及一種或多種翻化合物之藥學組 10成物可根據已知之藥學上有用組成物之製法調配。配方說明於熟諳技藝人士眾所周知且方便易得之多個來源。舉例言之’雷明頓製藥科學，默克出版公司，賓州伊士頓說明可關聯本發明使用之配方。適合投予之配方例如包括水性其可含有抗氧化劑、緩衝劑、10 Including a trichostatin compound and one or more suspension compounds may be combined with one or more pharmaceutically acceptable media and/or money = use. As used herein, "pharmaceutically acceptable medium" includes both complete and complete solutions, solvents, diluents, or other liquid vehicles, octagonal or suspending aids, surfactants, isotonic agents. Thickener paste, preservative, solid binder, riding agent, adjuvant, vehicle, sputum, sputum (four), absorbing (four), preservative, surfactant, scent, scent, or sweetness The solution is suitable for the desired dosage form ^ 15 20 = triclinic compound and one or more compounds which can be combined with excipients and optionally sustained release groups: The polymer combination is broken down to form a therapeutic composition. "Pharmaceuticals:: Agents" include non-toxic solid, semi-solid or liquid filters, tablets, encapsulating materials or formulation adjuvants of any type. The total use of the medical composition is based on the completeness of the clinician: the specific treatment for any particular treatment is effective:::: a number of factors, including, for example, the condition being treated and the specific composition of the condition & The activity of the substance; the age, weight, general health, sex and diet of the particular patient used; the music of the 'injection path; the rate of excretion of the specific compound used; the specific composition used in the treatment The combination of the use of the song 200831525 cilostatin compound and / or one or more platinum compounds is well known in the pharmaceutical industry. For example, it is known within the skill of the art to begin administering the composition at a lower dose than is required to achieve the desired therapeutic effect, and to slowly increase the dosage to achieve the desired efficacy. 5 Including a composition of a trichostatin compound and one or more platinum compounds For the administration of glutinous rice and uniformity of dosage, it is preferred to formulate a unit dosage form. "Unit dosage form" as used herein refers to a unit of separation on a body suitable for use in the composition of the host to be treated. Each dose must contain an amount of the composition calculated to bind to or in combination with the selected pharmaceutical carrier medium to produce the desired therapeutic effect. Preferred unit dosage formulations are unit dosage formulations containing the daily or permissible dosage unit, per unit dosage, or suitable component thereof. For example, a mother's day about 1_5 gram of the compound disclosed herein can reduce the volume of solid tumors in mice. The 15 doses will be determined based on factors such as body weight, age, surface area, metabolism, tissue distribution, rate of absorption, and rate of excretion. In one embodiment, from about 0.5 grams to about 7 grams of the triclinide compound as disclosed herein can be administered to humans. Optionally, from about 1 gram to 4 grams of triclinide compound on the mother's day can be administered to humans. In several embodiments, about 0.001 mg to 5 mg of troxaridin compound 20 per sputum can be administered to humans. The therapeutically effective dose will be determined based on a number of factors as described above. In addition, the composition can be administered at relatively low doses within the skill of the art, as well as increasing the dosage to achieve the desired effect. A composition comprising a trichostatin compound and one or more platinum compounds can be used for sustained release of the matrix, which can be made of a material that can be hydrolyzed by an enzyme or hydrolyzed based on acid, or by decomposition of a solution, usually a polymer. Once inserted into the body, the matrix is affected by _ and body fluids. The sustained release matrix is, for example, selected from biocompatible materials such as microbes, polylactide (polylactic acid), polyglycolic acid (glycolic acid polymer), and polyacetic acid vinegar copolymerized vinegar 5 ( Copolymer of lactic acid and glycolic acid), polyanhydrides, poly(orthoacid) vinegar, polysaccharides, hyaluronic acid, collagen, chondroitin sulfate, acid retardants, fatty acids, fats, polysaccharides, nucleic acids Classes, polyamino acids, amino acids such as phenylalanine, tyrosine, isoleucine m-acid, polyvinyl bromide, polyethylene glycol, and polychlorinated. Preferably, the biodegradable matrix is a matrix of one of polyacrylic acid (copolymer of lactic acid and glycolic acid). The triclinide compound and the one or more flavonoids can also be administered in the form of vesicles. As is known in the art, vesicles are typically derived from dish fats or other lipid materials. The vesicles are formed by dispersing a single layer or a plurality of layers of hydrated liquid crystal in an aqueous medium. Any non-toxic physiologically acceptable and metabolizable lipid which forms vesicles can be used. The vesicles may contain a stabilizer, a preservative, an excipient, etc., in addition to the composition or compositions of the present invention. Examples of lipids are natural and synthetic phospholipids and yue-based assays (India). The formation of microlipids is well known in the art world. 2〇 The compound and the uranium compound may be formulated as an aerosol to be applied, for example, by inhalation. These formulations for administration to the respiratory tract may be in the form of an aerosol or a spray solution or in the form of a powder for fine insufflation, either alone or in combination with an inert carrier such as a mortar. In this case, in one embodiment, the formulation particles have a diameter of less than 50 microns, less than 1 〇 micron in one embodiment of the 2008 200831525 embodiment. Compositions comprising a tricinem compound and one or more uranium compounds can be used in combination with other compositions and/or procedures for treating the aforementioned conditions. For example, tumor know-how is treated with a combination of surgery, radiotherapy or chemotherapy, one or more compositions of the invention, and one or more of the compositions of the invention can be administered to the patient to prolong the dormancy of the microtumor metastasis, and is stable. , inhibit or reduce the growth of any residual primary tumors. 7.1. Additional Examples A pharmaceutical composition comprising a triclinide compound and one or more compounds of the compounds can be formulated according to the known methods for preparing pharmaceutically useful compositions. The formulations are described in a variety of sources that are well known and readily available to those skilled in the art. For example, Remington Pharmaceutical Sciences, Merck Publishing Company, Pennsylvania Easton Description can be used in connection with the formulations used in the present invention. Formulations suitable for administration include, for example, water, which may contain antioxidants, buffers,

法較佳組合至少另一種治療方法，及非水性無菌注射溶液劑， 15 制菌劑、及讓配方變成與丨胞的生長，本發明方包括但非限於化學治 92 200831525 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，諸如投予抗癌劑。Preferably, the method combines at least one other method of treatment, and a non-aqueous sterile injectable solution, a bacteriostatic agent, and the formulation becomes a growth of cells, and the invention includes, but is not limited to, chemical treatment 92 200831525 treatment, radiotherapy, selection Sexually inhibit the treatment 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.

也可進行投予呈鹽之API-2(曲西立濱）。藥學上可接受 5之鹽之實例為與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，例如甲苯磺酸鹽、甲磺酸鹽、乙酸鹽、檸檬酸鹽、丙二酸鹽、酒石酸鹽、丁二酸鹽、笨曱酸鹽、抗壞血酸鹽、α -酮基戊二酸鹽、及α _甘油基磷酸鹽。也可形成適當之無機酸鹽包括硫酸鹽、硝酸鹽、重碳酸鹽、及 10 碳酸鹽。一，狀人〜溫吧^便用技藝界眾所周知之標準程序獲得，例如充分驗性化合物諸如胺與適當之形成生理上可接受之陰離子之酸反應。也可製造㈣之驗金屬（例如鈉、鉀或鋰）鹽或鹼土金屬（例如鈣）鹽。 15 20 曲西立濱化合物及-種❹自化合物可調配成藥學，卫成物且以多種制之投藥形式亦即經口或經腸道外，藉 Γ途徑、肌㈣徑、局部賴或皮下途射好個體諸如曰病人或生病動物。物了Γί:本發明之曲西立濱化合物及—種或多種麵化合 =且5樂學上可接受之媒劑（亦即载劑）諸如惰性稀釋劑 ==，_經系統性投藥。可封裝於硬殼或軟殼日月 =1成_彳或可直接與病人食物摻混。供以㈣’該等化合物組合—種❹種賦了攝食鍵劑、賴用鍵、喉片、膠囊劑、_、懸浮液劑、 93 200831525 製劑須含有至少可改變，方便地畺比之範圍。於 K為可獲得有效糖裝劑、糊劑等劑型使用。此等組成物及〇·ΐ/。活性劑。組成物及製劑之百分比當然係占給定單位劑型重量之約2 %至約6 0 %重 5 此種治療有用之組成物巾之活性化合物含劑量水平。劑諸如膠囊劑等也含有下列成分如一鈣，朋散劑諸如玉米澱粉、馬鈴薯资 10 :或:滑劑諸如硬脂酸鎂;及甜味劑諸如蔗;：果:藻:It is also possible to administer salt-producing API-2 (Quxi Libin). Examples of pharmaceutically acceptable salts of 5 are organic acid addition salts with acids which form physiologically acceptable anions, such as tosylate, methanesulfonate, acetate, citrate, C. Diacid salts, tartrates, succinates, succinates, ascorbates, alpha-ketoglutarate, and alpha-glyceryl phosphates. Suitable inorganic acid salts can also be formed including sulfates, nitrates, bicarbonates, and 10 carbonates. One, the person-like temperature is obtained by standard procedures well known in the art, for example, by fully testing a compound such as an amine with an appropriate acid forming a physiologically acceptable anion. It is also possible to manufacture (iv) a metal (e.g., sodium, potassium or lithium) salt or an alkaline earth metal (e.g., calcium) salt. 15 20 Quxilide compounds and compounds can be formulated into pharmacy, Weicheng and in a variety of forms of administration, that is, oral or parenteral, by means of sputum, muscle (4), local or subcutaneous Shoot individuals such as paralyzed patients or sick animals. : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : : It can be packaged in a hard shell or a soft shell. It can be directly mixed with the patient's food. (4) The combination of these compounds gives a feeding agent, a remedy, a throat tablet, a capsule, a suspension, and a preparation, and the formulation must contain at least a changeable, convenient ratio. It is used in K to obtain a dosage form such as an effective sugar preparation or paste. These compositions and 〇·ΐ/. Active agent. The percentage of the composition and formulation will, of course, be from about 2% to about 60% by weight of the given unit dosage form. 5 The active compound of such a therapeutically useful composition will contain a dosage level. Agents such as capsules and the like also contain the following ingredients such as calcium, a powder such as corn starch, potato 10: or a slip agent such as magnesium stearate; and a sweetener such as sugar cane;

當單劑諸如薄荷腦、冬綠油或樓桃口味制人士囊”除了前述類別之材料之外跃 =:::諸:r或聚乙二醇，有：體形狀。f5 4方式來修飾固體單位劑型的1 15或糖等/如’錠劑、丸劑或膠囊劑可以明劑、蠟、蟲| 、丨可含有本發明化When a single agent such as menthol, winter green oil or scented peach flavors, in addition to the materials of the aforementioned categories, jump =::: various: r or polyethylene glycol, have: body shape. f5 4 way to modify the solid Unit dosage form of 1 15 or sugar, etc. / such as 'tablets, pills or capsules can be used to modify the agent, wax, insects |

===味劑一味或〜且於其==:料_學上，入持續釋放製劑及裝置Γ，。此外，本發明化合物曲西立濱化合物及_籀藉注M r· 合物也可藉1 水中:2^或腹内投予。活性劑或其鹽之私見0可混合無毒製備。分散液飽 /液體聚乙二醇、三醋精、及其混合物製備q 94 200831525 製備。於尋常储存與使用條件下物生長之保藏劑。匕專衣劑含有防止微生 ==射或輸注之藥學劑型包括包含活性成分之 5 或水性分散液劑或無菌散劑，其適合臨時旱備無囷 >主射用溶液劑岑給要可㈣⑽t 溶液劑或分散液劑，視需要叮展封於微脂粒。總而古，曰件下必須為無菌、_二、^劑型於製造與儲存條 ' ㈨°液體戴劑或載媒劑可為溶劑刀政介質包括例如水、乙醇、多元醇(例如甘油、丙 10 15 =、液體聚乙二醇類等）、植物油、無毒甘㈣類、及其適虽混合物。適當流體性質例如可經由微脂粒之形成，經由於分散液之情況下_所“徑，或經由使用界面活性 Μ來、准持。藉各種抗囷劑及抗錢射獲得微生物作用的 Τ!:例如對經基苯甲酸醋類、氯丁醇、I山梨酸、柳瓜水等於夕種情況下，較佳含括等張劑例如糖類、緩衝劑或氯化納。注射驗成物可藉由於組成物巾使用延遲吸收劑例如—硬騎減a轉來獲得延長吸收。 20 無囷注射溶液劑之製法係經由將本發明化合物以需要，視需要於前述㈣成分摻辦適纽_，接著過渡滅菌於用於製備無菌注射用溶液劑之無菌散劑之情況下，製備方法為真轉狀冷料燥技術，獲料性成分加任可額外存在於@述經過無_過渡之溶液巾之期望成分之散劑。供局部投藥用，曲西立濱化合物及一種或多種翻化合物可以純質形歧用，亦即呈液體。但通常期望組合皮膚 95 200831525 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。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、铭氧等。有用之液體載劑包括水、醇類 5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最佳性質。所得液體組成物可由吸收墊施用，用於浸潰端帶及其它敷料，或使用幫浦型喷灑器或氣溶膠喷灑器而喷 10 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、及月曰肪酸酯類、脂肪醇類、改性纖維素類、或改性礦物材料也可用於液體載劑來形成可展開糊料、凝膠劑、軟膏劑、皂劑#供直接施用於使用者皮膚。可用來將本發明化合物 15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)、=== The scent is scented or ~ and in its ==: material _ learning, into the sustained release preparation and device Γ,. Further, the compound of the present invention, the trichostatin compound and the 籀借 borrowing M r compound, can also be administered by 1 in water: 2^ or intraperitoneally. The private agent of the active agent or its salt can be mixed and prepared non-toxic. Preparation of a dispersion of saturated/liquid polyethylene glycol, triacetin, and mixtures thereof prepared as q 94 200831525. Preservative for the growth of substances under normal conditions of storage and use.匕Specializing agent contains a pharmaceutical preparation containing microbial == injection or infusion, including an active ingredient 5 or an aqueous dispersion or a sterile powder, which is suitable for temporary drought and no sputum> Solution or dispersion, if necessary, spread on the liposome. In general, the sputum must be sterile, _2, ^ dosage form in the manufacture and storage of the '(9) ° liquid wear agent or vehicle can be a solvent knife medium including, for example, water, ethanol, polyol (such as glycerin, C 10 15 =, liquid polyethylene glycols, etc.), vegetable oils, non-toxic sweets (four), and their suitable mixtures. Appropriate fluid properties can be obtained, for example, via the formation of vesicles, via the dispersion, or via the use of interfacial activity, by means of various anti-caries agents and anti-money! For example, in the case of benzoic acid vinegar, chlorobutanol, I sorbic acid, and squash water, it is preferable to include an isotonic agent such as a saccharide, a buffer or a sodium chloride. The injection test can be borrowed. Since the composition towel uses a delayed absorbent, for example, hard riding minus a to obtain extended absorption. 20 The innocuous injection solution is prepared by adding the compound of the present invention to the above-mentioned (four) component as needed, and then In the case of the transitional sterilization in the case of a sterile powder for the preparation of a sterile injectable solution, the preparation method is a true-rotation cold-drying technique, and the material-accepting component is added to the expectation that the solution is additionally present in the non-transitioned solution. For the topical administration, the triconide compound and one or more compounds can be used in a pure form, that is, in a liquid form. However, it is generally desirable to combine the skin 95 200831525 acceptable carrier 'carriers can be The composition or formulation is applied to the skin as a solid or a liquid. Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, argon, oxygen, etc. Useful liquid carriers include water, alcohols 5 or a glycol or water-alcohol/glycol blend, wherein the compound of the invention may be dissolved or dispersed in an effective amount by means of a non-toxic surfactant as needed. Adjuvants such as perfumes and additional antimicrobial agents may be added. The best properties are obtained for a given application. The resulting liquid composition can be applied by an absorbent pad for dip end strips and other dressings, or sprayed onto the affected area using a pump spray or aerosol sprayer. Thickeners such as synthetic polymers, fatty acids, fatty acid hydrochloric acid, and montmorillonates, fatty alcohols, modified celluloses, or modified mineral materials can also be used in liquid carriers to form developable pastes. Materials, gels, ointments, soaps # for direct application to the skin of the user. Examples of useful skin compositions useful for delivering the compound 15 of the present invention to the skin are disclosed in Jacquet et al. Patent cases 4,608,392), Geria (US Patent 4,992,478),

Smith等人(美國專利案（娜⑸)及W()ltzman (美國專利案 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 20内之活性及於動物研究模型中之活體内活性來测定。於小鼠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。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(u_25wt_%或由約05_10wt_%: 一個 96 200831525 實施例中，於半固體組成物或固體組成物諸如凝膠劑或散劑中之濃度約為O.U wt··%，較佳約為0.5-2.5 Wt·-%。於一個實施例中，注射、輸注或攝食用之單一劑量通常為5-1500 毫克，可每曰投予μ3次來於成人獲得約0.1-50毫克/千克之 5濃度。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。Smith et al. (U.S. Patent (Na(5)) and W() ltzman (U.S. Patent 4,820,508). The useful dose of the pharmaceutical composition of the present invention can be compared by comparing its activity in test tube 20 with living organisms in animal research models. Intra-activity assays. Methods for extrapolation to humans in effective doses in mice and other animals are known in the art; for example, reference is made to U.S. Patent No. 4,938,949. In one non-limiting example, liquid compositions such as lotions The concentration of the active agent may be about (u_25wt_% or from about 05_10wt_%: in a 96 200831525 embodiment, the concentration in the semi-solid composition or solid composition such as gel or powder is about OU wt··%, Preferably, the dosage is 0.5-2.5 Wt·-%. In one embodiment, the single dose for injection, infusion or ingestion is usually 5 to 1500 mg, and may be administered 3 times per ounce to obtain about 0.1 to 50 mg per adult. The concentration of 5 in kilograms. 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.

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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一種或多種鉑化合物與藥學上可接受之載劑之組合。適合經口、經局部或經腸道外投藥之藥學組成物包括疋ΐ曲西立濱化合物及一種或多種翻化合物，該組成物組成本發明之較佳實施例。於本發明之上下文中，投予個體特別為人體之劑量為足夠於合理時間框影響病人的治療反應。熟諳技藝人士瞭解該劑量將依據多項因素決定，包括動物情況、動物體重及癌症之嚴重程度及癌症階段。適當劑量為於腫瘤組織獲得已知可發揮期望反應之曲西立濱化合物及-種舒雜化合物之濃度。較佳劑量為 =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。ΑΡΙ_2(或其藥學上可接受之鹽)之投予可為連續或以分開間隔投予，如技藝界熟諳技巧人士之判定。可由所揭示之抑制癌細胞生長方法獲益種包括但_於靈長_如猿、黑_、觀動物物居家動蝴如寵_如犬令天#、倉鼠、越南2 豬、兔、及雪紹；農場家畜諸如牛、水牛、野牛、馬諸、羊、及山羊；典型常見於動物園之珍奇異獸諸如7 97 200831525 獅、虎、豹、象、海馬、厚牛、長頸鹿、#羊、樹懶、燈羚二斑馬、牛羚、土撥鼠、無尾熊、袋鼠、負鼠、浣熊、書田能貨狗、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 5人類哺乳動物及非人哺乳動物物種。同理，試管試驗之本卷明方法可用於此種哺乳動物物種之細胞。需要使用本發明方法治療之病人可使用醫界專業人士已知之標準技術識別。 k供下列實例以供舉例說明但非限制性。 1〇 8·實例 8·1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或如前文說明（Cheng J.Q·等人，致癌基因，1997· 14: ρ·2793_2801; West Κ·Α·等人，今日抗藥性，2002. 5: 15 P.234-248; Satyamoorthy Κ·等人，癌症研究，2001· 61: P.7318-7324)。NCI結構分集集合為選自於約140,000化合物 NCI藥物倉儲中之1，992種化合物存庫。有關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NCI發展治療計劃網址。 20 經過Akt轉形細胞生長抑制之篩檢：經AKT2轉形之 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) 96一種溶液細胞增 98 200831525 - 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 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 10 Biotechnology Inc) ° 結果 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(JetztA.等人，癌症研究，2003. 63: 15 p.697-706, 2003)。如此，Akt被視為新穎癌症治療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 20 形細胞抑制生長。其中最強力之化合物API-2 (NCI識別符：NSC 154020)可於50 nM濃度遏止細胞生長。第1A圖顯示API-2也稱作為曲西立濱之化學結構式（Schweinsberg RD. 等人，Biochem Pharmacol，1981· 30: p.2521-2526)。API-2 選擇性抑制A K T- 2轉形細胞優於未經轉形之親代細胞的事 99 200831525 實，提醒發明人判定API-2是否為AKT2激_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 - 5 示，API-2於絲胺酸-309及絲胺酸-474(二者為AKT2完全活 - 化所需）顯著抑制AKT2麟酸化(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、 ^ 10 -AKT2及-AKT3處理，血清匱乏隔夜且以ΑΠ-2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）’提不八卩1-2不會於試管内直接抑 15 制Akt，API-2既非作為ATP競爭者，也非作為結合至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 ⑩ 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，由靶定Akt上游分子將可導 20 致API-2抑制Akt。因PI3K及PDK1為Akt之直接上游調節劑 (Datta S.R·等人，基因Dev，1999· 13: ρ·2905-2927)，檢驗 API-2是否抑制PI3K及/或PDK1。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-ρΐ 10 α抗體免疫沉澱。免疫沉澱物使 100 200831525 用PI-4-P作為酶基質接受試管pi3K激酶檢定分析。如第2A 圖所示，EGF-誘導之ΡΙ3Κ活性藉瓦特曼寧抑制但未受ΑΡΙ-2 抑制。欲評估ΑΡΙ-2對PDK1之效應，重組PDK1促進ΑΚΤ2 多月太之蘇胺酸-309磷酸化之檢定分析係於含磷脂基肌糖醇 - 5之脂質囊存在下使用。如第2Β圖所示，檢定分析藉對照組 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(5.1μΜ)，API-2只顯示21% 檢定分析之抑制。為了進一步評估ΑΡΙ-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 10 之PDK1之自行磷酸化程度，絲胺酸_241為自我磷酸化且對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）。 15 API-2 對透過PKC、PKA、SGK、STAT、JNK、p3820 Thus, the invention includes a pharmaceutical composition comprising a tricilide compound and one or more platinum compounds in combination with a pharmaceutically acceptable carrier. Pharmaceutical compositions suitable for oral, topical or parenteral administration include a tromethamine compound and one or more compounds which are preferred embodiments of the invention. In the context of the present invention, the dosage administered to an individual, particularly a human body, is a therapeutic response sufficient to affect the patient in a reasonable time frame. Skilled practitioners understand that the dosage will be determined based on a number of factors, including animal condition, animal weight and severity of cancer, and stage of cancer. The appropriate dose is to obtain the concentration of the trichostatin compound and the sedative compound which are known to exert the desired reaction in the tumor tissue. The preferred dose is = the maximum inhibition of cancer cell growth without the uncontrolled side effects. The administration of ΑΡΙ_2 (or a pharmaceutically acceptable salt thereof) can be administered continuously or at discrete intervals, as determined by skilled artisans. It can be benefited by the disclosed method for inhibiting the growth of cancer cells, including _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Farm animals such as cattle, buffalo, bison, horses, sheep, and goats; typical exotic animals commonly found in zoos such as 7 97 200831525 lions, tigers, leopards, elephants, seahorses, thick cows, giraffes, #羊,树Lazy, antelope two zebras, wildebeests, groundhog, koalas, kangaroos, possums, raccoons, books, dogs, seals, sea lions, walruses, otters, bottlenose dolphins, dolphins and whales. The terms "patient" and "individual" are used interchangeably herein and are intended to include 5 human mammalian and non-human mammal species. Similarly, the test method of the test tube can be applied to cells of such mammalian species. Patients in need of treatment using the methods of the present invention can be identified using standard techniques known to those skilled in the medical profession. k is provided by the following examples for illustration and not limitation. 1〇8·Example 8.1 Example 1: Intra-tube screening cell line and NCI diversity collection: All cell lines can be purchased from ATCC or as described above (Cheng JQ· et al., Oncogene, 1997·14: ρ· 2793_2801; West Κ·Α· et al., Drug Resistance Today, 2002. 5: 15 P.234-248; Satyamoorthy Κ· et al., Cancer Research, 2001· 61: P.7318-7324). The NCI structure diversity set is a library of 1,992 compounds selected from approximately 140,000 compound NCI drug stores. Further information on the selection, structure and activity of these diversity collection compounds can be found on the NCI Developmental Therapy Program website. 20 Screening for Akt-transformed cell growth inhibition: NIH3T3 cells transfected with AKT2 or NIH3T3 control cells transfected with LXSN vector (Cheng JQ· et al., Oncogene, 1997·14: ρ·2793·2801) Inoculate in a 96-well tissue culture well plate. After treatment with a 5 μΜ NCI diversity pooled compound, the cell growth line was assayed using CellTiter 96, a solution for cell growth (2008). 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 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 are purchased from Upstate 10 Biotechnology Inc. The result is the recognition of the small molecule inhibitor API-2 of the Akt signaling pathway. Human cancer detects Akt changes frequently, destroys the Akt pathway, induces Apoptosis and inhibition of tumor growth (Jetzt A. et al., Cancer Research, 2003. 63: 15 p. 697-706, 2003). As such, Akt is considered an attractive molecular target for the development of novel cancer therapeutics. 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-trans 20 cells. One of the most potent compounds, API-2 (NCI identifier: NSC 154020), inhibits cell growth at a concentration of 50 nM. Fig. 1A shows that API-2 is also referred to as the chemical structural formula of Tricinem (Schweinsberg RD. et al., Biochem Pharmacol, 1981 30: p. 2521-2526). API-2 selectively inhibits A K T-2 transfected cells over untransformed parental cells 99 200831525 Indeed, the inventors were reminded to determine whether API-2 is an AKT2 agonist. To achieve this, AKT2 was immunoprecipitated with anti-AKT2 antibody from NIHT3T3 cells transfected with akt-2 after treatment with API-2. AKT2 immunoprecipitation was performed using an anti-phospho-Akt antibody for immuno dot analysis. As shown in Figure 1B-5, API-2 significantly inhibits AKT2 linonic acidification in dextran acid-309 and serine-474 (both required for AKT2 complete viability) (Datta SR et al., Gene Dev , 1999· 13: ρ·2905-2927). Since the isoforms of the three Akt share a highly homologous, series, and similar structural formula, the effect of API-2 on its kinase activity was evaluated. HEK293 cells were treated with HA-Aktl, ^10-AKT2 and -AKT3 before stimulation with EGF (50 Ng/ml), serum was depleted overnight and treated with ΑΠ-2 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). -2 is neither an ATP competitor nor an enzyme matrix competitor that binds to the active site of Akt. API-2 does not inhibit known Akt upstream activators: it is well 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. Thus, by targeting a molecule upstream of Akt, it will be able to inhibit Akt by API-2. Since PI3K and PDK1 are direct upstream regulators of Akt (Datta S.R. et al., Gene Dev, 1999. 13: ρ. 2905-2927), it was examined whether API-2 inhibits PI3K and/or PDK1. HEK293 cells were depleted in serum and then treated with API-2 or PI3K inhibitor, Watmanin for 30 minutes prior to stimulation with EGF. PI3K was immunoprecipitated with an anti-ρΐ 10 α antibody. The immunoprecipitate was subjected to a test tube pi3K kinase assay using PI-4-P as an enzyme substrate. As shown in Figure 2A, EGF-induced Κ3Κ activity was inhibited by Watmanning but not by ΑΡΙ-2. To assess the effect of ΑΡΙ-2 on PDK1, the assay for recombinant PDK1 to promote 磷酸2 multi-monthly threonine-309 phosphorylation was used in the presence of a lipid vesicle containing phospholipid-based myositol-5. As shown in Figure 2, the assay was strongly inhibited by the control group PDK1 inhibitor staur〇Sp〇rine (ic5〇=5 ... nM). Conversely, at the highest test concentration (5.1 μΜ), API-2 only showed inhibition of 21% assay analysis. To further evaluate the effect of ΑΡΙ-2 on PDK1 activation, the degree of autophosphorylation of PDK1 in serine _241 10 was examined after ΑΡΙ-2 treatment with ΑΡΙ-2 cells, and serine _241 was self-phosphorylated. A residue of critical importance for activity (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). 15 API-2 pairs through PKC, PKA, SGK, STAT, JNK, p38

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 20 PKC相關激酶。於AGC激酶家族中，PKA、PKC及SGK之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ApU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。試管内激酶檢定分析及免疫墨點分析顯示PKA及 101 200831525 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Αρι_2對其活性不具影響（第2c圖及第圖）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 5癌基因存活徑路有影響。使用市售抗磷酸抗體之西方墨點分析顯示Stat3、JNK、p38及Erkl/2之磷酸化程度不受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 10胞生長且誘導細胞凋亡：API-2選擇性抑制Akt徑路之能力，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OVCAR3、OVCAR8、PANC1 及AKT2-15 轉形之 NIH3T3)或 PTEN 基因突變（PC-3、LNCaP、 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， 20 癌症研究，2001.6h ρ·7318-7324)。免疫墨點分析顯示只有於表現升高之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，ΑΠ-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A k t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、 200831525 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， • 5 或對藉IGF-1之生長刺激無反應。此外，API-2誘導細胞凋亡達 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 10 於表現失序Akt之細胞中抑制細胞生長及誘導細胞凋亡。 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 15 FKHRL1)、結節素（tuberlin)/TSC2、p70S6K、GSK-3 /5 、 p21WAF1/Cipl、p27kipl、MDM2、Bad、ASK1 及ΙΚΚα 等。其籲次檢驗API-2是否抑制Akt下游標靶。因抗-磷酸-結節素、 -Bad、-AFX、及-GSK-3石抗體為市面上可得，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 2〇後，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4Α圖顯示ΑΡΙ-2顯著抑制結節素之磷酸化程度，結果導致結節素之穩定與向上調節（Dan，H.C·等人，J Biol Chem, 2002.277: P.35364-35370)。Bad、GSK-3yS、及 AFX之磷酸化程度部分受API-2衰減。此等資料提示API-2 103 200831525 經由抑制其下游標靶之磷酸化來誘導細胞死亡及細胞生長停止。ΑΠ-2以不等程度抑制Akt下游標靶可能原因係在於此等標乾之填酸化位置也受其它激酶的調節，例如Bad絲胺酸-136除了藉Akt磷酸化之外也藉ΡΑΚΙ磷酸化（Schurmann， 5 Α·等人，Mol Cell Biol，2000.20: ρ·453-461)。 8·2實例2:於裸小鼠踵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 10 到約ioo-iso立方毫米時，動物經隨機分配，經每日腹内注射0·2毫升曲西立濱化合物及/或一種或多種鉑化合物之載媒劑。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。 ΑΡΙ_2於過度表現Akt之裸小鼠抑制腫瘤之生長：於人 15 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(SolitD.B. 20 等人，癌症研究，2003· 63: p.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: ρ·697-706, 2003)。因API-2抑制Akt發訊，且只於Akt濃度升高之癌細 200831525 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(OVCAR3、OVCAR8及PANC-1)皮下植 5 入右脅腹，而表現低度Akt之該等細胞系（〇乂〇八115及 C0L0357)則植入小鼠的左脅腹。當腫瘤達到平均大小約為二 100-150立方毫米時，動物隨機分組且使用載媒劑或API-2 (1¾克/千克/日）經腹内注射處理。如第4B圖所示，以載媒 _ 劑處理之OVCAR5及C0L0357腫瘤於腫瘤植入後49日生長Akt with ERK signaling pathway is highly selective: Akt belongs to the AGC ® (PKA/PKG/PKC) kinase family, which also includes PKA, PKC, serum-inducible kinase and glucocorticoid-inducible kinase (SGK), p90 Ribosomal S6 kinase, p70S6K, mitogen- and stress-activated protein kinases and 20 PKC-associated kinases. In the AGC kinase family, the protein structural formula of PKA, PKC and SGK is closer to the protein structure of Akt kinase than other members. Therefore, the effect of ApU on the catalytic activity of the three kinases was examined. HEK293 cells were transfected with HA-tagged PKA, PKCa or SGK. In vitro assay and immunoblotting analysis showed that the kinase activities of PKA and 101 200831525 PKC α were inhibited by a PKC inhibitor, namely PKAI and R〇31-8220, respectively; while Αρι_2 had no effect on its activity (Fig. 2c and Figure VII) In addition, serum-inducible SGK kinase activity is attenuated by Wattmannin 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 blot analysis using commercially available anti-phospho antibodies showed that the degree of phosphorylation 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. Api_2 inhibits fine cell growth and induces apoptosis in Akt overexpressed/activated human cancer cell lines: the ability of API-2 to selectively inhibit Akt pathways, suggesting that this is due to a disordered performance/activation of Akt The tumor cells inhibit proliferation and/or induce apoptosis. API-2 is used to treat overexpression of AKT2 (NIH3T3 transduced with OVCAR3, OVCAR8, PANC1 and AKT2-15) or PTEN gene mutations due to Akt overexpression or PTEN mutations leading to activation of Akt in human malignancies ( PC-3, LNCaP, 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., 20 Cancer Research, 2001.6h ρ·7318-7324). Immunoblotting analysis showed that the degree of phosphorylation of Akt was inhibited by API-2 only in cells with elevated Akt or response to IGF-1 stimulation (Fig. 3A). Thus, ΑΠ-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 overexpression/activated cell line, 200831525 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, • 5 or did not respond to growth stimulation by IGF-1. In addition, API-2 induced cell apoptosis 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 10 to inhibit cell growth and induce apoptosis in cells exhibiting disordered Akt. 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 15 FKHRL1), tuberlin/TSC2, p70S6K, GSK-3 /5, p21WAF1/Cipl, p27kipl, MDM2, Bad, ASK1, and ΙΚΚα. It recalled whether API-2 inhibited Akt downstream targets. Since anti-phospho-nodulin, -Bad, -AFX, and -GSK-3 stone antibodies are commercially available, the effect of API-2 on phosphorylation by Akt was determined. After treatment with API-2 (ΙμΜ) for 2〇, OVCAR3 cells were lysed and immunoblot analysis was performed using individual anti-phospho antibodies. Figure 4 shows that ΑΡΙ-2 significantly inhibits the degree of phosphorylation of nodulin, resulting in the stabilization and upregulation of nodulin (Dan, H. C. et al., J Biol Chem, 2002. 277: P. 35364-35370). The degree of phosphorylation of Bad, GSK-3yS, and AFX is partially attenuated by API-2. These data suggest that API-2 103 200831525 induces cell death and cell growth arrest by inhibiting phosphorylation of its downstream targets. The possible reason for the inhibition of Akt downstream targets by ΑΠ-2 is also regulated by other kinases in the position of this standard dry filling. For example, Bad serine-136 is phosphorylated by Akt phosphorylation. (Schurmann, 5 Α· et al., Mol Cell Biol, 2000.20: ρ·453-461). 8.2 Example 2: Antitumor activity in a nude mouse tumor xenograft study model Tumor cells were harvested, suspended in PBS, and subcutaneously injected into the right and left flank of a 8 week old female nude mouse as previously reported. (2χ1〇6 cells/flank) (Sun J. et al., Cancer Research, 1999. 59: ρ·4919-4926, 1999). When the tumor reaches 10 to about ioo-iso cubic millimeters, the animals are randomly assigned to inject 0.2 ml of the troxaribine compound and/or one or more platinum compound carriers daily. Control animals received DMSO (20%) vehicle, while treatment animals were injected with ΑΠ-2 (1 mg/kg/day) in 20% DMSO. ΑΡΙ_2 inhibits tumor growth in nude mice overexpressing Akt: shows frequent overexpression/activation and/or expansion of AKT1 and AKT2 in human ovarian cancer and pancreatic cancer (Cheng JQ· and Nicosia SV·, in cancer) Encyclopedia Reference, 2001, Schwab D., Editing Springer, Berlin, Heidelberg and New York, pp. 35_37). Akt pathway is inhibited by inhibitors such as PI3K, HSP70, Src and farnesyltransferase, leading to cell growth arrest and induction of apoptosis (Solit D. B. 20 et al., Cancer Research, 2003. 63: p. 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: ρ·697-706, 2003). Because API-2 inhibits Akt signaling and induces apoptosis and cell growth arrest only in cancer cells with elevated Akt concentrations (Fig. 3), tumor growth with elevated Akt content should be compared to nude mice. Tumor growth in tumors with lower Akt levels in the body is more sensitive to API-2. In order to achieve this, subcutaneous injection* Akt overexpressing cells (OVCAR3, OVCAR8, and PANC-1) were subcutaneously implanted into the right flank, and these cell lines exhibiting low Akt (〇乂〇八115 and C0L0357). Then implanted into the left flank of the mouse. When tumors reached an average size of approximately two hundred-150 cubic millimeters, animals were randomized and treated with vehicle or API-2 (13⁄4 g/kg/day) by intra-abdominal injection. As shown in Figure 4B, OVCAR5 and C0L0357 tumors treated with vehicle-based agents grew on day 49 after tumor implantation.

10 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 15 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。 20 8·3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/或蘇胺酸-308。於含有磷脂基肌糖醇-3,4,5-P3 (PIP3)、API-2及 105 200831525 組織呋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。 8.4實例4 : TCN於癌症抗性細胞中有效 5 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。 8.5實例5:於西鉑汀抗性卵巢癌細胞中TCN克服西鉑汀抗性 · 10 對西鉑汀敏感之卵巢癌細胞系、A27 8 0 S及OV200 8及其西鉑汀抗性衍生株、A2780CP及C13分別以0、10、20&30μΜ 西鉑汀處理，接著評估西鉑汀處理存活細胞百分比。於 A2780CP及C13細胞系驗證對西鉑汀毒性之抗性（第9Α 圖）。得自全部四種細胞型之溶解產物藉SDS-PAGE分割，且以抗磷酸化Akt及總Akt之抗體探測。西鉑汀抗性細胞系内生性表現顯著較高濃度之礙酸化Akt (第9B圖）。其次評估TCN對西麵汀抗性產生何種影響。A2780CP Φ 及C13卵巢細胞系以〇、5、10及20μΜ含或未含ΐ〇μΜ TCN 處理，接著測定細胞存活率。添加TCN顯著降低二細胞系 2〇對西鉑汀之抗性（第1〇圖）。然後C13細胞以〇、丨、5、10及 20μΜ TCN含或未含1(¥厘西鉑汀處理，接著測定細胞存活率。TCN與西鉑汀的組合可協同性降低細胞存活率（第u 圖）〇 C13細胞於裸小鼠體内皮下注射，接著以載媒劑 106 200831525 (DMSO)、單獨西鉑汀(2毫克/千克/日）、單獨TCN(2毫克/千克/日）、或TCN及西鉑汀處理。以每週間隔評估腫瘤質量共計4週。西鉑汀與TCN之組合物可提升遏止腫瘤生長及進行的能力（第12圖）。 5 8·6實例6 : TCN提升mTOR抑制效果 ΟV3細胞及MCF7細胞以mTOR抑制劑亦即拉帕徽素及 RAD001分別處理經24小時時間。溶解產物藉Sds-PAGE切10 to about 800-1000 cubic millimeters. 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 15 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. In the treated tumor samples, Akt activity was affected by API-2, 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. 20 8·3 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 threonine-308. Intracellular kinase assay analysis of recombinant strains of PDK1 and Akt in a kinase buffer containing phospholipid-based inositol-3,4,5-P3 (PIP3), API-2 and 105 200831525 tissue furan H2B as enzyme substrate . After 30 minutes of incubation, the reaction was separated by SDS-PAGE and exposed to the membrane. 8.4 Example 4: TCN is effective in cancer-resistant cells. The effect of 5 TCN (API-2) is on West Platinum, Pacific Paclitaxel, and Tasmania, Moxifen resistance A270CP, C-13, OVCAR433, and MCF7/TAM. 'Cellular test. In these cells, API-2 overcomes the resistance of cetamine, paclitaxel, alcohol, and tamoxet. 8.5 Example 5: TCN overcoming cisplatin resistance in cisplatin-resistant ovarian cancer cells · 10 ovarian cancer cell lines sensitive to citratetin, A27 80 S and OV200 8 and their western platinum-resistant derivatives A2780CP and C13 were treated with 0, 10, 20 & 30 μΜ xiplatin, respectively, and then the percentage of viable cells treated with xiplatin was evaluated. Resistance to West Platinum toxicity was verified in the A2780CP and C13 cell lines (Fig. 9). Lysates from all four cell types were digested by SDS-PAGE and probed with antibodies against phosphorylated Akt and total Akt. The western platinum-resistant cell line exhibits a significantly higher concentration of acid-accumulating Akt (Fig. 9B). Secondly, it is evaluated how the effect of TCN on the resistance of the Westing. The A2780CP Φ and C13 ovarian cell lines were treated with 〇, 5, 10, and 20 μM with or without ΐ〇μΜ TCN, followed by cell viability. The addition of TCN significantly reduced the resistance of the two cell line 2〇 to the western platinum (Fig. 1). Then C13 cells were treated with 〇, 丨, 5, 10, and 20μΜ TCN with or without 1 (? cisplatin, and then cell viability was determined. The combination of TCN and citrate can synergistically reduce cell viability ( u Figure 〇C13 cells were injected subcutaneously into nude mice, followed by vehicle 106 200831525 (DMSO), citrate alone (2 mg/kg/day), TCN alone (2 mg/kg/day), or Treatment with TCN and citrate. The tumor mass was assessed at weekly intervals for a total of 4 weeks. The combination of citrate and TCN increased the ability to suppress tumor growth and progression (Fig. 12). 5 8·6 Example 6: TCN boost mTOR inhibitory effect ΟV3 cells and MCF7 cells were treated with mTOR inhibitors, ie, rapamycin and RAD001, for 24 hours. The lysate was cut by Sds-PAGE.

割且移轉至膜上接受西方墨點分析。膜以抗填酸_Akt、總 Akt、磷酸-p70S6K及總p70S6K之抗體探測。帶有衍生自〇V3 10 細胞浴解產物之膜額外使用抗礙酸-FKHRL1及總FKHRL1 之抗體探測。mTOR抑制劑的處理結果導致於兩個細胞系中 Akt徑路之快速活化（第13圖）。然後OV3細胞以對照組、拉帕黴素、或拉帕黴素及處理；MCF7細胞以對照組、RAD001或{^^⑻丨及丁^^處 I5理。TCN顯著降低mTOR抑制劑所導致之处鱗酸化升高而未隨後造成P70S6K活性的增高（第14圖）。然後於對照組、 mTOR抑制劑、TCN、或TCN及mTOR抑制劑存在下，評估 DU-145、MCF7及OV3細胞生長速率經6曰時間。藉mT〇R 抑制對細胞生長之抑制作用可藉TCN提升（第15圖）。 20 8.7實例7 : TCN克服奥羅拉誘導之西鉑汀抗性奥羅拉-A為涉及細胞週期進行之絲胺酸/蘇胺酸激酶。檢驗奥羅拉_八對於對西鉑汀敏感度的影響。Α278〇§及 A2780CP細胞以及OV2008細胞以空白載體轉移感染，或以編碼經HA加標籤之奥羅拉-a蛋白質之組成體轉移感染。奥 107 200831525 羅拉_A之表現可經由西方墨點分析經由SDS_pAGE切割之溶解產物且轉移至膜獲得驗證（第16圖，上圖）。細胞存活率係於太平洋紫杉醇及西鉑汀存在下評估。以奥羅拉_A轉移感采之A2780S及OV2008細胞顯示對以只使用空白載體轉 5和感染之A2780S細胞及OV2008細胞改良之存活率（第16 圖）。其次評估奥羅拉-A對西鉑汀誘導細胞凋亡之影響。 A2780S細胞以空白載體轉移感染或以編碼奥羅拉_A蛋白貝之構成體轉移感染。然後細胞以西翻汀處理。免疫螢光 10染色（第17A圖）顯示奥羅拉-A之表現可抑制胞色素c之釋放。西方墨點分析顯示導入奥羅拉-A可提高Akt之磷酸化及 Akt激酶活性（第17B圖及第17C圖）。其次以空白載體或奥羅拉-A轉移感染之A2780S及 OV2008細胞以西鉑汀、TCN或西鉑汀及TCN處理。表現奥 15羅拉-A之A2780S細胞及OV2008細胞皆顯示於TCN存在下對西鉑汀之抗性降低（第18圖）。已經參照較佳實施例說明本發明。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。 20 【闽式簡單說明】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。A顯示ΑΠ-2 (曲西立濱）之化學結構式。B顯示API-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 200831525 ΑΡΙ-2 (ΙμΜ)處理指示之時間，且接受以抗磷酸_Akt_T3〇8 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。C顯示API-2抑制Akt之三個同質異形體。 HEK293 細胞於 EGF 刺激前以 jjA-Aktl、HA-AKT2、及 5 HA-AKT3轉移感染且以Αρι_2 (1μΜ)或瓦特曼寧 (wortmannm)(15pM)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。D顯示API-2於試管 10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 ΑΡΙ-2 (線道3)。第2圖驗證ΑΡΙ-2不會抑制pI3K、PDK1及agc激酶家族中之緊密相關成員。A顯示試管内PI3K激酶檢定分析。於 EGF刺激前’ ΗΕΚ293細胞經血清匱乏且以API-2 (1 μΜ)或瓦 I5特又f 處理30分鐘。細胞經溶解且以anti_pi 1〇 q抗體免疫沉澱。免疫沉澱物使用PI_4-P作為酶基質接受活體内激酶檢定分析。B顯示API-2對試管内PDK1活化之效應（上圖），實心圓顯示藉API-2抑制。空心圓顯示藉陽性對照史妥洛史寶靈（staurosporine)抑制，史妥洛史寶靈為強力PDKl 20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-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。^ 顯示PKC α以抗構酸-PKC α _T638(上圖）及總PKC α (下圖）抗體接著以ΑΡΙ-2或非選擇性PKC抑制劑Ro31 -8220處理之 109 200831525 磷酸化程度之免疫墨點分析。D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 5之HA_SGK之表現。E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（ΑΠ-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ρ中顯示西方墨點。OVCAR3細胞以ΑΠ-2處理指示時間。細胞 10 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 (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凋亡。A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 15 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）。於B，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 (CellTiter) 96細胞增生檢定分析套件組（普米嘉公司 (Promega))分析。C提供細胞凋亡分析。細胞以API-2處理， 20 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A，驗證API-2可抑制結節素(tuberin)、Bad、AFX及GSK-3 召之Akt磷酸化。於以API-2處理後，OVCAR3細胞經溶解 200831525Cut and transfer to the membrane for Western blot analysis. Membranes were probed with antibodies against acid-Akt, total Akt, phospho-p70S6K and total p70S6K. Membranes with a deuterated product derived from 〇V3 10 cells were additionally probed with antibodies against acid-FKHRL1 and total FKHRL1. Treatment with mTOR inhibitors resulted in rapid activation of the Akt pathway in both cell lines (Fig. 13). The OV3 cells were then treated with the control group, rapamycin, or rapamycin; MCF7 cells were treated with the control group, RAD001 or {^^(8) 丨 and 丁^^. TCN significantly reduced the increase in serotonation caused by mTOR inhibitors without subsequently causing an increase in P70S6K activity (Fig. 14). The growth rates of DU-145, MCF7 and OV3 cells were then assessed over 6 曰 in the presence of control, mTOR inhibitor, TCN, or TCN and mTOR inhibitors. The inhibition of cell growth by mT〇R inhibition can be enhanced by TCN (Fig. 15). 20 8.7 Example 7: TCN Overcomes Aurora-Induced West Platinum Resistance Aurora-A is a serine/threonine kinase involved in the cell cycle. The effect of Aurora _8 on the sensitivity to West Platinum was examined. Α278〇§ and A2780CP cells and OV2008 cells were transfected with a blank vector or infected with a composition encoding the HA-tagged Aurora-a protein. AO 107 200831525 The performance of Rolla_A can be verified by Western blot analysis of the lysate cut by SDS_pAGE and transferred to the membrane (Fig. 16, upper panel). Cell viability was assessed in the presence of paclitaxel and cetidin. The A2780S and OV2008 cells, which were induced by Aurora-A transfer, showed improved survival rates of A2780S cells and OV2008 cells using only blank vector transfer 5 and infection (Fig. 16). Secondly, the effect of Aurora-A on the apoptosis induced by xiplatin was evaluated. A2780S cells were infected with a blank vector or infected with a construct encoding the Aurora-A protein shell. The cells are then treated with Westing. Immunofluorescence 10 staining (Fig. 17A) shows that the presence of Aurora-A inhibits the release of cytochrome c. Western blot analysis showed that the introduction of Aurora-A increased Akt phosphorylation and Akt kinase activity (Fig. 17B and Fig. 17C). Secondly, A2780S and OV2008 cells infected with blank vector or Aurora-A were treated with western platinum, TCN or cetylplatin and TCN. Both A2780S cells and OV2008 cells exhibiting O. 15 roller-A showed reduced resistance to c-platin in the presence of TCN (Fig. 18). The invention has been described with reference to the 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. 20 [Simple description of the 闽] Figure 1 shows the identification of API-2 (Quxi Libin) identified as a candidate for Akt inhibitors in the NCI diversity set. A shows the chemical structure of ΑΠ-2 (Quxi Libin). B shows that API-2 inhibits the degree of AKT2 phosphorylation in AKT2 transduced NIH3T3 cells. Wil-type AKT2_transformed NIH3T3 cells were treated with 200831525 ΑΡΙ-2 (ΙμΜ) for the indicated time and were subjected to immunoblotting analysis with anti-phospho-Akt_T3〇8 and -S473 antibodies (top and bottom) . The figure below shows the performance of the total AKT2. C shows that API-2 inhibits three isoforms of Akt. HEK293 cells were infected with jjA-Aktl, HA-AKT2, and 5 HA-AKT3 prior to EGF stimulation and treated with Αρι 2 (1 μΜ) or wortmannm (15 pM), and the cells were lysed and immunoprecipitated with anti-HA antibody. Immunoprecipitates were subjected to in vitro assays for kinase assays (top) and immunosorbent assays with anti-phosphoric acid-Akt-T308 (bottom) antibodies. The middle panel shows the performance of Aktl, AKT2, and AKT3 transfected. D shows that API-2 does not inhibit Akt in tube 10. The kinase assay for the active AKT2 recombinant protein in a test tube in a kinase buffer containing 1 μΜ ΑΡΙ-2 (lane 3). Figure 2 demonstrates that ΑΡΙ-2 does not inhibit closely related members of the pI3K, PDK1 and agc kinase families. A shows in vitro PI3K kinase assay analysis. Before EB stimulation, ΗΕΚ293 cells were serum-deficient and treated with API-2 (1 μΜ) or watt I5 and f for 30 minutes. The cells were lysed and immunoprecipitated with an anti_pi 1〇 q antibody. The immunoprecipitate was assayed for in vivo kinase assay using PI_4-P as the enzyme substrate. B shows the effect of API-2 on PDK1 activation in vitro (top panel), and the filled circle shows inhibition by API-2. The open circles showed inhibition by the positive control history of Sturosporine, which was a potent PDKl 20 inhibitor (IC50 = 5 nM). The following figure shows the immunoblotting of HEK293 cells. HEK293 cells were infected with Myc-PDK1 prior to EGF stimulation and treated with Watmanin or API-2. The immune dots were detected with the indicated antibodies. ^ Show PKC α with anti-acid-PKC α _T638 (top panel) and total PKC α (lower panel) antibody followed by ΑΡΙ-2 or non-selective PKC inhibitor Ro31-8220. 109 200831525 Immunological ink Point analysis. D shows in vitro SGK kinase assay analysis. HEK293 cells were infected with HA-SGK transfer prior to EGF stimulation and treated with API-2 or Watmanning. 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 after metastasis 5 . E shows the results of the PKA kinase assay. The immunopurified PKA was cultured in ADB buffer (Upstate Biotechnology Inc) containing the appropriate inhibitor (ΑΠ-2 or PKAI) and the enzyme substrate Kemptide. Quantify kinase activity. Show western ink dots in ρ. OVCAR3 cells were treated with ΑΠ-2 for indicated time. Cell 10 lysate was immunized with the appropriate anti-phospho antibody (1-4) and anti-actin antibody (bottom panel). Figure 3 demonstrates that API-2 induces apoptosis in Akt-expressing human cancer cells by inhibiting Akt activity and cell growth. A is a Western blot after treatment with API-2, and the degree of phosphorylation of Akt is detected by an anti-phospho-Akt-T308 antibody 15 in a suitable human cancer cell line. The ink spots were again probed with anti-total Akt antibodies (bottom panel). At B, a cell proliferation assay was performed. 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 CellTiter 96 Cell Proliferation Assay Kit (Promega). C provides apoptosis analysis. Cells were treated with API-2, 20 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. At A, validation of API-2 inhibits Akt phosphorylation by tuberin, Bad, AFX, and GSK-3. After treatment with API-2, OVCAR3 cells were dissolved 200831525

且以適用之抗體進行免疫墨點分析。B顯示API-2可抑制腫瘤生長。腫瘤細胞注射入裸小鼠體内，左侧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 5 理。各個測量值表示10個腫瘤之平均值。C顯示帶有以API-2 或載媒劑（對照組)處理之OVCAR3 (右）及OVCAR5 (左）之小鼠之代表圖。D顯示於實驗結束時之腫瘤大小（下）及重量（上) 之實例。E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OVCAR_3-衍生 10 腫瘤中之抗磷-Akt-S473 (上）及抗-AKT2 (下）抗體進行。第5圖顯示API-2 (曲西立濱)抑制於試管内抑制激酶活性。試管内激酶檢定分析係使用PDK1與Akt之重組株於含磷酸基肌糖醇-3,4,5-P3 (PIP3)、API-2及組織呋H2B作為酶基質之激酶緩衝液進行。培養30分鐘後，藉SDS-PAGE 15 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。第7a-7d圖提供人Akt2之mRNA及胺基酸序列，也標示限剪酶位置。 20 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圖顯示西翻汀（cisplatin) (CDDP)抗性細胞系内生性表現超活性Akt。第9A圖顯示兩種卵巢癌細胞系及OV2008)及以遞增量西麵汀處理之其西顧、汀抗性對偶（分 111 200831525 別為A2780CP及C13)之線圖。西鉑汀抗性變異株亦即 A2780CP及C13驗證回應於西鉑汀之存活率增高。第9B圖顯示西翻汀敏感性卵巢癌細胞亦即A2780S及OV2008與其西翻江抗性對偶之比較。上圖顯示於西鉑汀抗性細胞系C13 5 及A2780S中之超活性Akt。下圖顯示於各類細胞中存在之 Akt總量。第10圖顯示曲西立濱（TCN) (API-2)可克服西鉑汀抗性。西翻汀抗性細胞系亦即A2780S及C13係以0、5、10及 20μΜ西翻、;丁處理。白色條紋柱表示只以西鈷汀處理。黑灰 10 柱表示以西鉑汀與ΙΟμΜ TCN處理。以TCN共同處理可降低細胞存活率，指示TCN可克服西鉑汀抗性。第11圖顯示使用T C Ν及西鉑汀對C13卵巢癌細胞之細胞存活率所見協同效應。C13細胞以〇、1、5、10及2〇μΜ TCN 含及未含ΙΟμΜ西鉑汀處理。西鉑汀與TCN之組合物可協同 15 性地降低細胞存活率。第12圖顯示TCN可提升西鉑汀抑制於裸小鼠異種移植片中C13卵巢癌細胞生長之能力。裸小鼠接受C13異種移植片，接著以載媒劑(DMSO)、單獨西鈾、;丁、單獨tcn或西翻 >丁與TCN處理’接著分析於接種後7、14、21、或28日時之 20腫瘤退行。左圖（A)顯示一線圖驗證西鉑汀與TCN之組合物顯著降低腫瘤之生長。右圖（B)顯示於接受代表性處理之接種小鼠體内之腫瘤質量。第13圖顯示於人卵巢癌細胞中Akt徑路藉mT〇R抑制劑之快速活化。上圖（A)顯示於使用1 nMmT〇R抑制劑拉帕黴 200831525 素(rapamycin)處理指示之時間後，〇V3細胞溶解產物之西方墨點分析。膜係以對磷酸化同基因型及對總蛋白質具有特異性之抗體探測用於Akt、p70S6K及FKHRL 1之比較。下 ‘ 圖（B)顯示於使用mTOR抑制劑RAD001處理指定之時間 • 5 後，MCF7細胞溶解產物之西方墨點分析。膜係以對填酸化同基因型及對總蛋白質具有特異性之抗體探測用於Akt及二 P70S6K之比較。第14圖顯示透過mTOR抑制劑AKT徑路之活化係藉 _ TCN衰減。左圖顯示於OV3細胞中TCN對於以拉帕黴素處 10 理之Akt磷酸化之效應。右圖（B)顯示於MCF7細胞中TCN對以RAD001處理之Akt磷酸化之相同效應。第15圖顯示經由TCN與RAD001或拉帕徽素之組合對細胞生長之效應提升。左上圖（A)為線圖，顯示以對照組、單獨拉帕黴素、單獨TCN或TCN與拉帕黴素處理6曰， ^ 15 DU_145細胞之細胞數目。右上圖（B)為線圖，顯示以對照組、單獨RAD001、單獨TCN或TCN與RAD001處理6曰， • MCF7細胞之細胞數目。下圖（C)為線圖，顯示以對照組、單獨拉帕黴素、單獨TCN或TCN與拉帕黴素處理6日，OV3 細胞之細胞數目。 20 第16圖顯示經由奥羅拉(Aurora)-A活化Akt誘導細胞存活及化學抗性。上圖（A)顯示於A2780S、A2780CP及OV2008 印巢癌細胞系中奥羅拉_A成功地轉移感染及表現。第二圖 (B)為線圖顯示於八27808細胞中表現奥羅拉-八提高細胞存活率及對太平洋紫杉醇(paclitaxel)之抗性。第三圖（C)為線 113 200831525 圖顯示於A2780S細胞中表現奥羅拉-A提高細胞存活率及對西鉑汀之抗性。下圖（D)顯示於OV2008細胞中表現奥羅拉-A提南細胞存活率及對西舶彡丁之抗性。第17圖顯示奥羅拉-A對胞色素c釋放及Akt活化之效 5 應。第ΠΑ圖顯示於使用空白載體或使用或未使用西鉑汀處理之奥羅拉-A轉移感染之A2780S細胞中胞色素c之免疫螢光。西鉑汀誘導胞色素c的釋放受奥羅拉-A表現之抑制。第 17B圖為西方墨點顯示奥羅拉-A之表現可提高於西麵汀敏感細胞中之Akt之磷酸化。第17C圖顯示提高奥羅拉_A濃度 10 可增加Akt之磷酸化及活化。第18圖顯示TCN(API-2)對奥羅拉_A誘導西鉑汀抗性之效應。上二圖（A及B)顯示以空白載體或奥羅拉-A轉移感染，接著單獨使用西鉑汀、單獨API-2或API-2及西鉑汀處理之A2780S細胞。下二圖（C及D)顯示以空白載體或奥羅拉 15 -A轉移感染，接著單獨使用西鉑汀、單獨API-2或API-2及西鉑汀處理之OV2008細胞。TCN降低於A2780S及OV2008 兩種細胞中經由奥羅拉-A表現所提供之西鉑汀抗性。第19圖顯示API-2、奥羅拉-A、p53及Akt於細胞存活所扮演之角色之示意圖。 20 第20圖顯示Src家族酪胺酸激酶抑制劑雷沙堤尼 (dasatinib)(史普利索（Sprycel))對八375細胞中之8代填酸化及Akt填酸化之效應。Src酪胺酸激酶之抑制可提高Akt之磷酸化。【主要元件符號說明】 (無） 114Immunoblot analysis was performed with applicable antibodies. B 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 reaches an average size of about 100-150 cubic millimeters, the animal is treated with vehicle or at 1 mg/kg/day API-2. Each measurement represents the average of 10 tumors. C shows a representative of mice with OVCAR3 (right) and OVCAR5 (left) treated with API-2 or vehicle (control). D shows examples of tumor size (bottom) and weight (top) at the end of the experiment. In E, the immunoblot analysis of tumor lysates used anti-phosphorus-Akt-S473 in OVCAR_3-derived 10 tumors treated with API-2 (T3 and T4) and untreated (T1 and T2) (top) And anti-AKT2 (lower) antibodies were performed. Figure 5 shows that API-2 (Cucidine) inhibits in vitro inhibition of kinase activity. The in-vitro kinase assay was performed using a recombinant strain of PDK1 and Akt in a kinase buffer containing phospho-inositol-3,4,5-P3 (PIP3), API-2 and tissue furose H2B as an enzyme substrate. After 30 minutes of incubation, the reaction was separated by SDS-PAGE 15 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. Figures 7a-7d provide mRNA and amino acid sequences of human Akt2, also indicating the position of the restriction enzyme. 20 Figure 8a-8c provides the mRNA and amino acid sequence of human Akt3, also indicating the position of the restriction enzyme. Figure 9 shows that the cisplatin (CDDP) resistant cell line endogenously exhibits superactive Akt. Fig. 9A shows a line graph of two ovarian cancer cell lines and OV2008) and their treatment with an increasing amount of cisplatin, which is treated with a serotonin and a serotonin-resistant dual (sub. 111 200831525, A2780CP and C13). The West Platinidine-resistant variants, namely A2780CP and C13, demonstrated an increase in survival rate in response to Western Platinum. Figure 9B shows a comparison of the sensitivity of diarrhea-sensitive ovarian cancer cells, namely A2780S and OV2008, to their resistance to Xijiangjiang. The top panel shows superactive Akt in the western platinum-resistant cell lines C13 5 and A2780S. The figure below shows the total amount of Akt present in various types of cells. Figure 10 shows that Tricinem (TCN) (API-2) overcomes the resistance of West Platinum. The anti-resistant cell lines of A. sinensis, namely A2780S and C13, were treated with 0, 5, 10 and 20 μΜ; White striped bars indicate that only Cobaltine is treated. The black ash 10 column indicates treatment with cisplatin and ΙΟμΜ TCN. Co-treatment with TCN reduced cell viability and indicated that TCN overcomes the resistance of cetyltin. Figure 11 shows the synergistic effect of cell survival on C13 ovarian cancer cells using T C Ν and cepacidine. C13 cells were treated with 〇, 1, 5, 10, and 2〇μΜ TCN with and without ΙΟμΜxiplatin. The combination of cisplatin and TCN can synergistically reduce cell viability. Figure 12 shows that TCN enhances the ability of coptin to inhibit the growth of C13 ovarian cancer cells in nude mouse xenografts. Nude mice received C13 xenografts, followed by vehicle (DMSO), uranium alone, butyl, tcn alone or diced with TCN and then analyzed 7, 14, 21, or 28 after inoculation. 20 tumors of the day regressed. The left panel (A) shows a one-line plot to verify that the combination of West Platin and TCN significantly reduced tumor growth. The right panel (B) shows the tumor mass in the recipient mice receiving representative treatment. Figure 13 shows the rapid activation of the Akt pathway by mT〇R inhibitor in human ovarian cancer cells. The above figure (A) shows the western blot analysis of the lysate of 〇V3 cells after the time indicated by the treatment of 1 nMmT〇R inhibitor Laptomycin 200831525. Membrane lines were probed for comparison of phosphorylated isoforms and antibodies specific for total protein for Akt, p70S6K and FKHRL 1. Bottom ‘ Figure (B) shows Western blot analysis of MCF7 cell lysates after treatment with the mTOR inhibitor RAD001 for a specified time • 5 . Membrane lines were probed for the comparison of Akt and P70S6K with antibodies that were acidified isoforms and specific for total protein. Figure 14 shows that the activation of the AKT pathway through the mTOR inhibitor is attenuated by _ TCN. The left panel shows the effect of TCN on Akt phosphorylation with rapamycin in OV3 cells. The right panel (B) shows the same effect of TCN on Akt phosphorylation treated with RAD001 in MCF7 cells. Figure 15 shows the effect of cell growth on the combination of TCN with RAD001 or lapala. The upper left panel (A) is a line graph showing the number of cells treated with the control group, laparomycin alone, TCN alone or TCN and rapamycin at 6曰, ^ 15 DU_145 cells. The upper right panel (B) is a line graph showing the number of cells in the MCF7 cells treated with the control group, RAD001 alone, TCN alone or TCN and RAD001. Figure (C) below is a line graph showing the number of cells in OV3 cells treated with control, laparomycin alone, TCN alone or TCN and rapamycin for 6 days. 20 Figure 16 shows activation of Akt via Aurora-A to induce cell survival and chemoresistance. The above figure (A) shows that Aurora_A successfully transferred infection and performance in the A2780S, A2780CP and OV2008 Insect Cancer Cell lines. The second panel (B) is a line graph showing the presence of Aurora-Augmented cell survival rate and resistance to paclitaxel in eight 27808 cells. Figure 3 (C) is line 113 200831525 The figure shows that Aurora-A enhances cell viability and resistance to cetillin in A2780S cells. Figure (D) below shows the survival rate of Aurora-A Tynan cells and the resistance to G. mellifera in OV2008 cells. Figure 17 shows the effect of Aurora-A on cytochrome c release and Akt activation. The panel is shown in immunofluorescence of cytochrome c in A2780S cells infected with a blank vector or with Aurora-A metastasis treated with or without Western Platinum. The release of cytochrome c induced by citrate was inhibited by Aurora-A. Figure 17B shows that Western blots show that the performance of Aurora-A can increase the phosphorylation of Akt in the Westin-sensitive cells. Figure 17C shows that increasing Aurora _A concentration 10 increases phosphorylation and activation of Akt. Figure 18 shows the effect of TCN (API-2) on Aurora-A-induced resistance to citrate. The above two panels (A and B) show A2780S cells treated with blank vector or Aurora-A transfer, followed by West Platin, API-2 or API-2 alone, and West Platinum. The next two panels (C and D) show infection with a blank vector or Aurora 15-A, followed by cisplatin alone, API-2 alone or API-2 and cisplatin-treated OV2008 cells. TCN was reduced in West Platinidine resistance provided by Aurora-A performance in both A2780S and OV2008 cells. Figure 19 shows a schematic representation of the role of API-2, Aurora-A, p53, and Akt in cell survival. 20 Figure 20 shows the effect of the Src family tyrosine kinase inhibitor dasatinib (Sprycel) on acidification and Akt acidification in eight generations of eight 375 cells. Inhibition of Src tyrosine kinase increases phosphorylation of Akt. [Main component symbol description] (none) 114

Claims

200831525 十、申請專利範圍： 1. 一種組成物，包含： ⑴式I-IV化合物：200831525 X. Patent application scope: 1. A composition comprising: (1) a compound of formula I-IV:

CHiCHi

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）；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 115 200831525 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尺2,、&，或R, 5 分別為Η或一磷酸、二磷酸、或三碟酸；Wherein 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); decylamine, sulfonate includes alkyl or arylalkyl; sulfonyl includes methanesulfonyl and benzyl, wherein the phenyl can be used as needed One or more 115 200831525 substituted 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 the rule 2, &, or R, 5 is hydrazine or monophosphate, diphosphate, or trioctate;

其中R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石買醯基，脂質包括>5粦脂質；胺基酸；破水化合物；胜 1〇肽，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、或環狀烷基（包括低碳烷基）、烯基或炔基、C〇_烷基、co-烯基、co_炔基、C0-芳基或雜芳基、c〇_烧氧 15 基烷基、C〇-芳氧基烷基、CO_經取代之芳基、磺醯基、烧基續醯基、芳基磺醯基、芳烷基磺醯基；Wherein R and Ry are each hydrogen, optionally substituted phosphate; sulfhydryl (including lower sulfhydryl); decylamine, alkyl (including lower alkyl); aromatic, deoxyalkylene such as poly Ethylene glycol, optionally substituted aryl sulphate, sulphonate including > 5 粦 lipid; amino acid; water-breaking compound; 〇 1 〇 peptide, or cholesterol; or other pharmaceutically acceptable leaving group; In one embodiment, the compound is administered as a 5,-phosphorus ether lipid or a 5,-ether lipid; each of Ri and R2 is a linear, branched, or cyclic alkyl group, respectively, which may be substituted, optionally including Lower alkyl), alkenyl or alkynyl, C〇-alkyl, co-alkenyl, co-alkynyl, C0-aryl or heteroaryl, c〇_calcinyl 15 alkyl, C〇- An aryloxyalkyl group, a CO-substituted aryl group, a sulfonyl group, a decyl group, an arylsulfonyl group, an aralkylsulfonyl group;

(H)如式V之一種或多種|自化合物：(H) one or more of formula V | from the compound:

式V 其中： Ri、R2、R3及R4各自分別為氫、視需要可經取代之胺、芳香族胺、雜芳香族胺、視需要可經取代之直鏈、 116 20 200831525 分支或環狀烷基、R!、R2、R3及R4間所形成之芳香環、雜芳香環或環狀環； R5及R6各自分別為鹵素、視需要可經取代之烷氧 ' 基、視需要可經取代之酯、視需要可經取代之烷基胺、 ^ 5 芳香族胺、雜芳香族胺； (iii)藥學上可接受之載劑。二 2.如申請專利範圍第1項之組成物，其中該式I化合物為曲西立濱（triciribine)。 # 3.如申請專利範圍第1項之組成物，其中該式I化合物為磷 10 酸曲西立濱。 4. 如申請專利範圍第1項之組成物，其中該式I化合物為磷酸曲西立濱一水合物。 5. 如申請專利範圍第1項之組成物，其中該式V鉑化合物為Wherein: Ri, R2, R3 and R4 are each independently hydrogen, optionally substituted amines, aromatic amines, heteroaromatic amines, optionally substituted straight chain, 116 20 200831525 branched or cyclic alkane An aromatic ring, a heteroaromatic ring or a cyclic ring formed between R, R, R3 and R4; each of R5 and R6 is a halogen, optionally substituted alkoxy group, optionally substituted An ester, optionally substituted alkylamine, ^5 aromatic amine, heteroaromatic amine; (iii) a pharmaceutically acceptable carrier. 2. The composition of claim 1, wherein the compound of formula I is triciribine. # 3. The composition of claim 1, wherein the compound of formula I is tromethamine. 4. The composition of claim 1, wherein the compound of formula I is triclinbium phosphate monohydrate. 5. The composition of claim 1 wherein the platinum compound is

%%

其中： Ri、R2、R3及R4各自分別為氫、視需要可經取代之胺、芳香族胺、雜芳香族胺、視需要可經取代之直鏈、分支或環狀烧基、Ri、R2、R3及R4間所形成之芳香環、雜芳香環或環狀環；化5及於6各自分別為鹵素。 6.如申請專利範圍第1項之組成物，其中該一種或多種鉑化合物係選自於由下列化合物所組成之組群： 117 20 200831525Wherein: Ri, R2, R3 and R4 are each hydrogen, optionally substituted amine, aromatic amine, heteroaromatic amine, optionally substituted linear, branched or cyclic alkyl, Ri, R2 An aromatic ring, a heteroaromatic ring or a cyclic ring formed between R3 and R4; each of 5 and 6 is a halogen. 6. The composition of claim 1, wherein the one or more platinum compounds are selected from the group consisting of: 117 20 200831525

ο aο a

... , "ΐ —3 ψ% ρι^η2 叫 m$ 叫... , "ΐ —3 ψ% ρι^η2 is called m$

7. 如申請專利範圍第1項之組成物，其中該式I化合物係以至少10毫克/平方米之劑量存在。 8. 如申請專利範圍第1項之組成物，其中該式V化合物係以 5 至少1毫克/平方米之劑量存在。7. The composition of claim 1, wherein the compound of formula I is present in a dose of at least 10 mg/m 2 . 8. The composition of claim 1, wherein the compound of formula V is present at a dose of 5 at least 1 mg/m 2 .

9. 如申請專利範圍第1項之組成物，其係適合供經腸道外投予。 10. 如申請專利範圍第9項之組成物，其中該腸道外投予為經靜脈投予。 10 11.如申請專利範圍第1項之組成物，其係適合供經口投予。 12. 如申請專利範圍第1項之組成物，其係適合供局部投予。 13. —種於一哺乳動物治療腫瘤或癌症之方法，包含對該哺乳動物投予有效量之一種組成物包含： ⑴式I-IV化合物： 118 2008315259. If the composition of claim 1 is suitable for parenteral administration. 10. The composition of claim 9, wherein the parenteral administration is intravenous administration. 10 11. The composition of claim 1 is suitable for oral administration. 12. If the composition of claim 1 is suitable for topical administration. 13. A method of treating a tumor or cancer in a mammal, comprising administering to the mammal an effective amount of a composition comprising: (1) a compound of formula I-IV: 118 200831525

其中R2、RS及Rs’分別為氫、視需要可經取代之磷酸根或膦酸根（包括一碟酸、二碟酸、或三麟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、磺酸酯包括烧基或芳基燒基；磺醯基包括甲磺基及卞基，其中違本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；视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 119 200831525 5 10 15 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，_填醚脂質或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、或環狀烷基（包括低碳烷基）、烯基或炔基、CO-烷基、CO-烯基、CO-炔基、CO-芳基或雜芳基、CO-烷氧基烷基、CO-芳氧基烷基、CO-經取代之芳基、磺醯基、烧基磺醯基、芳基磺醯基、芳烷基磺醯基；以及 (ii)如式V之一種或多種鉑化合物：Wherein R2, RS and Rs' are respectively hydrogen, optionally substituted phosphate or phosphonate (including one-plate acid, two-disc acid, or tri-seminic acid or stabilized phosphoric acid prodrug); mercapto group (including low carbon) Alkyl (including lower alkyl); decylamine, sulfonate including alkyl or arylalkyl; sulfonyl including methylsulfonyl and fluorenyl, which may be subjected to a 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 from 119 200831525 5 10 15 de-based, which provides a compound in vivo wherein R 2 , R 3 , or R 5 are respectively hydrazine or monophosphate, diphosphate, or triphosphate; Ry is hydrogen, optionally substituted sulphate; sulfhydryl (including lower sulfhydryl); decylamine, alkyl (including lower alkyl); aromatic, deoxyalkylene such as polyethylene glycol An arylsulfonyl group which may be substituted as needed; a lipid including a phospholipid; an amino acid Carbohydrate; peptide; or cholesterol; or other pharmaceutically acceptable leaving group; in one embodiment, the compound is administered as a 5,_ether ether lipid or a 5,_ether lipid; Ri and R2 are each直, optionally substituted straight chain, branched, or cyclic alkyl (including lower alkyl), alkenyl or alkynyl, CO-alkyl, CO-alkenyl, CO-alkynyl, CO-aryl Or heteroaryl, CO-alkoxyalkyl, CO-aryloxyalkyl, CO-substituted aryl, sulfonyl, alkylsulfonyl, arylsulfonyl, aralkyl sulfonate a sulfhydryl group; and (ii) one or more platinum compounds of formula V:

式V 其中： Ri、R2、R3及R4各自分別為氫、視需要可經取代之胺、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、、R2、R3及R4間所形成之芳香環、雜芳香環或環狀環； R5及R0各自分別為鹵素、視需要可經取代之燒氧 120 20 200831525 基、視需要可經取代之酯、視需要可經取代之烷基胺、芳香族胺、雜芳香族胺。 14.如申請專利範圍第13項之方法，其中式I-IV化合物及式 V化合物係同時投予。 - 5 15.如申請專利範圍第13項之方法，其中式I-IV化合物經投予後，接著投予式V化合物。二 16.如申請專利範圍第13項之方法，其中式V化合物經投予後，接著投予式I-IV化合物。 • 17.如申請專利範圍第13項之方法，其中該式I-IV化合物係 10 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。 18.如申請專利範圍第13項之方法，其中該式V化合物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。 — 15 19.如申請專利範圍第18項之方法，其中該投藥計劃係重複至少兩次。 ® 20.如申請專利範圍第18項之方法，其中該投藥計劃係重複至少四次。 21. 如申請專利範圍第13項之方法，其中該接受治療之腫瘤 20 為***腫瘤、胰臟腫瘤、卵巢腫瘤及大腸直腸腫瘤。 22. 如申請專利範圍第13項之方法，其中投予至少10毫克/ 平方米式I-IV化合物。 23. 如申請專利範圍第13項之方法，其中投予100毫克/平方米或以下之式I-IV化合物。 121 200831525 24. 如申請專利範圍第13項之方法，其中投予至少1毫克/千克式V化合物。 25. 如申請專利範圍第13項之方法，其中投予10毫克/千克或以下之式V化合物。Wherein: Ri, R2, R3 and R4 are each independently hydrogen, optionally substituted amines, aromatic amines, heteroaromatic amines, optionally substituted linear, branched or cyclic alkyl groups, An aromatic ring, a heteroaromatic ring or a cyclic ring formed between R2, R3 and R4; each of R5 and R0 is a halogen, optionally substituted calcined oxygen 120 20 200831525 base, optionally substituted ester, depending on A substituted alkylamine, an aromatic amine, or a heteroaromatic amine is required. 14. The method of claim 13, wherein the compound of formula I-IV and the compound of formula V are administered simultaneously. The method of claim 13, wherein the compound of the formula I-IV is administered, followed by the administration of the compound of the formula V. A method of claim 13, wherein the compound of formula V is administered, followed by administration of a compound of formula I-IV. 17. The method of claim 13, wherein the compound of formula I-IV is administered once a week for three weeks, followed by one week of administration of no compound. 18. The method of claim 13, wherein the compound of formula V is administered once a week for three weeks, followed by one week of no administration of any compound. — 15 19. The method of claim 18, wherein the dosing plan is repeated at least twice. ® 20. The method of claim 18, wherein the dosing schedule is repeated at least four times. 21. The method of claim 13, wherein the treated tumor 20 is a breast tumor, a pancreatic tumor, an ovarian tumor, and a colorectal tumor. 22. The method of claim 13, wherein at least 10 mg/m 2 of the compound of formula I-IV is administered. 23. The method of claim 13, wherein a compound of formula I-IV of 100 mg/m2 or less is administered. 121 200831525 24. The method of claim 13, wherein at least 1 mg/kg of the compound of formula V is administered. 25. The method of claim 13, wherein a compound of formula V of 10 mg/kg or less is administered.

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