TW200831111A - Compositions including triciribine and anthracyclin analogs and methods of use thereof - Google Patents

Abstract

This application relates to combination therapies including triciribine and related compounds and anthracycline analogs and salts thereof and compositions with reduced toxicity for the treatment and prevention of tumors, cancer, and other disorders associated with abnormal cell proliferation.

Description

。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 FIELD OF THE INVENTION The present application relates to low toxicity combination therapies suitable for the treatment and prevention of tumors, cancer, and other disorders associated with abnormal cell proliferation, including tricinel and related compounds and Onioncycline analogues and their salts, '10%. [Prior Art 1 BACKGROUND OF THE INVENTION Cancer is an abnormal growth of cells. Despite the limitations of space, the nutrients shared by other sputum or the body's signal to stop the regeneration, the cancer regenerates at a speed of 15 speeds. The shape of cancer cells is usually different from healthy cells, and its function is

Normal and can spread into many parts of the body. Abnormal tissue: A tumor, called a tumor, is a population of cells that can grow uncontrollably and divide. The tumor can be bipolar (non-cancerous) or malignant (cancerous). Benign tumors tend to be slow to produce ... and + will govern. Malignant tumors can grow rapidly, invade and destroy, normal -20 and can spread throughout the body. 'The classification of cancer is based on the body part from which the fluid or tissue or root from which it originates. In addition, some cancers are mixed. Cancer can be classified into five types: generalized types: carcinoma, sarcoma, lymphoma, leukemia, and myeloma, and the types of cancers and blood groups (4). Carcinomas are tissues found in the body 5 200831111, that is, the epithelial tissues that cover the surface of organs, glands or body structures or as the lining of them. For example, a cancer of the stomach lining is called a cancer. Many cancers affect organs or glands involved in secretion, such as the mammary glands that produce breast milk. Cancer is the main cause of about 80 to 90% of all cancer cases. Sarcomas are 5 self-connecting tissues, such as cartilage, fat, muscle, tendon, and bone, and malignant tumors that grow. The most common sarcoma, a tumor on the bone, usually exists in young people. Examples of sarcomas include osteosarcoma (bone) and chondrosarcoma (cartilage). Lymphoma refers to a cancer that originates from the knot or gland of lymphocytes, which functions to produce white blood cells and purify body fluids, or organs such as the brain and breast. Lymphomas fall into two categories: 10 Hodgkin's lymphoma and non-Hodgkin's lymphoma. Leukemia, also known as blood cancer, is a bone marrow cancer that prevents the bone marrow from producing 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 negative blood. Platelets prevent the body from easily stabbing and bleeding. Examples of leukemia include acute myeloid leukemia, chronic bone white leukemia, acute lymphocytic leukemia, and chronic lymphocytic leukemia. The terms "myeloid" and "lymphocyte" refer to the type of cell involved. Finally, the myeloma line grows in the plasma cells of the bone marrow. In some cases, the myeloma cells can accumulate in a bone and form a single tumor, i.e., a cell tumor. In other cases, however, the myeloma cells may accumulate within the 20th multi-bone, thereby forming a bone tumor, also known as a multi-myeloma. Tumor induction and evolution are usually the result of changes in the accumulation of the tumor cell genome. Such changes may include inactivation of a cell growth inhibitory gene or a tumor suppressor gene, and activation of a cell growth promoting gene or oncogene. Many activated cell oncogenes have been identified in animal models to date, 200831111, and only a few of these genes have been shown to be associated with human cancer (Weinberg), 19 9 Oncogenes and the Molecular Origins of Cancer Cold Spring Harbor, NY, Stanbridge and Nowell 1990 Cell 63 867-874, Godwin et al., 1992 Oncogenes and antioncogenes in 5 gynecological malignancies. In WJ Hoskins, CA Perez and RC Young (eds), Gynecological oncology principles and Rarhce, pp 87-116, Lippincott, Philadelphia). The activation of oncogenes in human cancer can result from factors such as increased gene replication or structural changes. These factors can lead to many cellular actions, such as their ability to cause overexpression of gene products. Several oncogenes involved in human cancer can be activated via gene overexpression. It is clear that continuous genetic abnormalities in cancer cells lead to defects in regulatory signal transduction circuits that manage normal cell proliferation, differentiation, and programmed cell death (Hanahan, D. and RA Weinberg, Cell, 2000. 15 1〇〇 ( 1): Ρ· 57-700). This in turn leads to a fundamental defect in cell physiology, which can lead to malignancy. These defects include: a) self-sufficiency of growth signals (ie, growth factor receptor tyrosine acid, such as EGFR overexpression and downstream signal transduction pathways such as Ras/Raf/Mek/Erk% and Ras/PI3K/Akt Abnormal activation), b) resistance to growth signals (ie, low expression of 2〇TGFB and its receptors), c) escape from apoptosis (ie loss of pro-apoptotic p53; pro-survival Bcl- Excessive performance of 2; survival pathways, such as the high activation of the survival pathway of the media by P13K/Akt, continuous angiogenesis (ie, high secretion of VEGF) and f) tissue invasion and metastasis (ie, extracellular proteases and protoplasts) Metastatic integrin) (Hanahan, D. and RA 7 200831111

Weinberg, Cell, 2000·100(1): ρ· 57-700) 0 Receptor tyrosine kinases, such as EGFR, ErbB2, VEGFR and insulin-like growth factor I receptor (IGF-1R), are closely involved in many humans Cancer, which includes the formation of colorectal cancer, pancreatic cancer, breast cancer, and ovarian cancer 5 (Khaleghpour, K., et al, Carcinogenesis, 2004. 25(2)····················· Human, Cancer Res, 2003. 63(22): ρ· 7708-16) 〇 ligands, such as EGF, VEGF and IGF-1, bind to their receptors to promote the stimulation of endogenous tyrosine kinase activity The autophosphorylation of the action and specific enzymes in the somatic pulp domain of these receptors and the recruitment of signaling proteins that trigger 10 complex signal transduction pathways (Olayioye, MA, et al, Embo J , 2000· 19(13): ρ·3159-67, Porter, AC and RR Vaillancourt, Oncogene, 1998·17 (11 Reviews): p· 1343-52). This in turn leads to many tumor viability and tumorigenic pathways, such as the activity of these Ras/Raf/Mek/Erk%, JAK/STAT3 and PI3K/Art pathways. 15 Although all three pathways are involved in tumorigenesis involving the colon, pancreas, breast, and ovary, these pathways by Akt have been shown to be malignant, including cell proliferation, anti-apoptosis/survival, invasion, and metastasis. It is important in many steps with angiogenesis (Datta, SR et al. Genes Dev, 1999. 13(22): ρ. 2905-27). 20 Akt is a serine/threonine protein kinase (also known as guanidine) with three family members: AkU, Akt2 and Akt3. Stimulate cells with growth or survival factors to recruit lipid kinase, phosphoinositide-3-OH-kinase (PI3K), which phosphorylates phosphoinositide-4,5-diphosphate (PIP2) to form PIP3, which can help The cell membrane is supplemented with Akt, in which Akt can be activated by the acidification reaction 200831111 of Thr308 and Ser473 (Aktl), Thr308 25 and Ser474 (Akt2) and Thr308 and Ser472 (Akt3) (Datta, SR· et al. Genes Dev, 1999) , 13(22): ρ· 2905-27). Therefore, PI3K can activate Akt by phosphorylating PIP2 and converting it to PIP3. The phosphatase, guanidine, dephosphorylates ΡΙΡ3 to form ΡΙΡ2, thus preventing Akt activation. 5 Most human cancers contain highly activated Akt (Datta, S.R. et al.

Genes Dev, 1999·13(22): ρ· 2905-27, Bellacosa, A·, et al, Int J Cancer, 1995·64(4): ρ. 280-5; Sim, M·, et al, Am J Pathol, 2001. 159(2): p. 431-7). In more detail, Akt 10 is over-expressive and/or highly active in 57%, 32%, 27%, and 36% of human colorectal cancer, pancreatic cancer, breast cancer, and ovarian cancer, respectively (Roy, Η.K , et al. Carcinogenesis, 2002·23(1): ρ. 201·5·, Altomare, DA·, et al, J Cell Biochem, 2003.88(l): p. 470-6·, Sun, M·, etc. People, Cancer Res,

2001.6l(16): p. 5985-91·, Stal, O” et al. Breast Cancer Res, 2003· 5(2): p· R37-44, Cheng. J·Q·, et al., Proc Natl Acad Sci 15 USA, 1992· 89(19): p. 9267-71, Yuan, ZQ·, et al., Oncogene, 2000· 19(19): p. 2324-30). The high activation of Art is caused by Akt itself. Amplification and/or overexpression and genetic alterations upstream of Akt, including overexpression of receptor tyrosine kinases and/or their ligands (Khaleghpour, K., et al. Carcinogenesis, 2004. 25(2) )·_ρ· 241_8; 20 Sekharam, Μ·, et al, Cancer Res, 2003. 63(22): ρ· 7708-16, Cohen, BD·, et al, Biochem Soc Symp, 1998· 63: ρ· 199 -210·, Muller, WJ· et al. Biochem Soc Symp, 1998. 63: ρ· 149-57, Miller, WE” et al. J Virol, 1995·69(7): ρ· 4390-8, Slamon, DJ” Et al., Science, 1987. 235 (4785): p. 177-82, Andmlis, IL, 25 et al, J Clin Oncol, 1998. 16(4): p· 1340-9) for the phosphatase PTEN 9 200831111 Deletion. It has been proved that the ectopic expression of Akt induces malignant deformation and promotes cell survival (Sun, M” et al. Am J Pathol, 2001· 159(2): ρ· 431-7, Cheng, J.Q” et al., 〇ncogene, 1997·14(23): ρ· 2793-801) and

Division of the Akt pathway inhibits cell growth and induces apoptosis (Jetzt, Α·, 5 et al. Cancer Res, 2003· 63(20): ρ·6697-706) and preclinical instructions

Akt is involved in proof of concept for tumor formation. Current treatments for cancer and related diseases are limited in effectiveness and have many serious unintended side effects. Although many anticancer drugs have proven to be clinically effective, severe systemic toxicity usually halts clinical development of 10 successful chemotherapeutic agents. In addition, overexpression of receptor tyrosine kinases (such as EGFR) and their ligands (such as IGF-1), excessive expression and/or loss of Akt in PTEN (which all lead to high activation of Akt) and cancer The patient's poor prognosis, resistance to chemotherapy, and shortened survival. Current research strategies emphasize the search for effective treatments with low risk. 15 The dose-limiting toxicity and chemical resistance of onionine analogs can force optimization of therapeutic therapies. Therefore, the synergistic effect of the combination of the trichostatin compound and the onioncycline analog can be used as a combination therapy for treating tumors, cancer, and abnormal cell proliferation. SUMMARY OF THE INVENTION 3 In one embodiment, the present invention provides a combination of triclinide, tricineridine phosphate and related compounds with onioncycline to treat a tumor or cancer in a patient and to limit systemic toxicity Novel treatment options. The present invention is based on the discovery that tumors or cancers that overexpress Akt kinase are particularly sensitive to the potentiating effects of cells of TCN and related compounds 200831111 and the synergistic effects produced by the use of onioncycline analogs. In contrast to prior art and experience, the inventors have determined successful methods of using triclinide and onioncycline analogs to treat tumors and cancer by one or a combination of the following: (i) to Quxili The compounds of the compound and the onion 5 cyclin analogs exhibit enhanced sensitivity are administered to the drug; (ii) the toxicity of the triclinide compound and/or the onionine analog can be minimized but still Said dosage which shows efficacy; or (iii) the use of said administration regimen which minimizes the toxicity of the triclinide compound and/or the onionine analog. In an illustrative embodiment of the invention, the invention encompasses a composition comprising: (1) a compound of formula I to IV or a combination thereof:

11

2008311H

Wherein R3' and R5' are each independently hydrogen, optionally substituted phthalic acid or a phosphonate (which includes a mono-, di- or triphosphate or a stabilized phosphate 5); It includes a lower fluorenyl group; an alkyl group (which includes a lower alkyl group); a decylamine, a sulfonate including an alkyl group or an aralkyl group; a sulfonyl group including a sulfonium group and a sulfhydryl group, wherein The phenyl group may be optionally substituted with one or more substituents as described, for example, in the definition of aryl groups as taught herein; optionally substituted arylsulfonyl; lipids, including phospholipids; amino acids; a carbohydrate compound; a peptide; or cholesterol; or other pharmaceutically acceptable cleavage group in which R2, 113, and 115 are independently a compound of hydrazine or mono-, di- or tri-phosphate, wherein Rx and Ry are independently hydrogen, optionally substituted phosphate; sulfhydryl (which includes lower sulfhydryl); decylamine, alkyl (which includes lower alkyl); aromatic 15 polyoxyalkylene, such as poly Ethylene glycol, optionally substituted arylsulfonyl; lipids, including phospholipids; amino acids; carbohydrates Peptide; or cholesterol; or other pharmaceutically acceptable exfoliating group. In one embodiment, the 12 200831111 compound is administered as a 5,-phosphate ether lipid or a 5,_ether lipid;

Ri and R2 are each independently Η, optionally substituted straight chain, branched chain or % alkyl group (which includes lower alkyl group), alkenyl or alkynyl group, c〇-alkyl group, c〇_ alkenyl group , CO-alkynyl, CO-aryl or heteroaryl, c〇-alkoxyalkyl, c〇_aryloxyalkyl, CO-substituted aryl, sulfonyl, alkanesulfonyl, arylsulfonate a aryl sulfonyl group; (ii) an analog of the formula V or an anthracycline of the formula VI,

Wherein the compound of formula VI has the structure:

10 type V

Ri, R4, R6 and R7 are each independently hydrogen, hydroxy and alkoxy; R2 and R3 are each independently hydrogen, alkyl, alkoxy, light, _; R5 is hydrogen, optionally substituted alkane a base chain, a branched chain or a cyclic alkyl group, a hydroxyl group, an alkoxy group; 15 Rs is a CO-alkyl group, a CO-halogen-substituted alkyl group, a CO-aryl group or a heteroaryl group; and wherein the compound of the formula VI has the following structure : 13 200831111

The core is independently hydrogen, alkyl, alkoxy, hydroxy, halogen; r3 and r4 are each independently hydrogen, optionally substituted alkyl chain, branched 5-chain or cyclic alkyl, optionally a substituted alkyl chain, a branched chain or a cyclic alcohol, an optionally substituted alkyl chain, a branched chain or a cyclic amine; and (m) a pharmaceutically acceptable carrier. In another illustrative embodiment, the invention encompasses a method of treating a tumor or cancer in a mammal comprising administering to the mammal an effective amount of a composition comprising less than 10 compounds: (1) a compound of formulas I to IV or Combination:

14 200831111

Wherein 2', R3' and R5' are each independently hydrogen, optionally substituted phosphate or phosphonate (which includes mono-, di- or tri-phosphate or stabilized phosphate prodrugs); (which includes a lower fluorenyl group); an alkyl group (which includes a lower alkyl group); a decylamine, a terephthalate, which includes an alkyl or aralkyl group; a sulfonyl group, which includes a methylsulfonyl group and a benzyl group, Wherein the phenyl group is optionally substituted with one or more substituents as described, for example, in the definition of an aryl group as taught herein; a selectively substituted aromatic acid group; a lipid comprising a phospholipid; an amine group Acid; carbohydrate

1°, peptide; or cholesterol; or R2, in vivo. Other pharmaceutically acceptable, cleavable groups of compounds which are either hydrazine or mono-, di- or tri-phosphate; wherein Rx and V are independently hydrazine, optionally substituted phosphate; thiol (which includes low Carbenyl); decylamine, alkyl (which includes lower alkyl); aromatic 15 oxime to oxidize olefins, such as polyethylene glycol, optionally substituted arylsulfonyl 'barium' which includes phospholipids; Amino acids; carbohydrates; peptides; or cholesterol, or other pharmaceutically acceptable exfoliating groups. In one embodiment, the 15 200831111 compound is administered as a 5'-linoleate lipid or a 5,-ether lipid; in a particular embodiment, the core and each are independently H, a selectively substituted straight chain , branched or cyclic alkyl (which includes lower alkyl), alkenyl or alkynyl, CO-alkyl, CO-alkenyl, C0_fast, CO-aryl or heteroaryl, 5 C ( > alkoxyalkyl, c〇-aryloxyalkyl, co-substituted aryl, sulfonyl, alkanesulfonyl, arylsulfonyl, aralkylsulfonyl; and indium ring of formula V or formula VI A analogue wherein the compound of formula VI has the structure:

10 type V

R1, R4, 尺6, and R7 are each independently hydrogen, a trans group, and an alkoxy group; R2 and R3 are each independently hydrogen, an alkyl group, an alkoxy group, a sulfhydryl group, a halogen group; and R5 is a hydrogen group, which is optionally substituted. a base bond, a branched or a ring alkyl group, a mesogenic group, an alkoxy group; 15 R8 is a CO-alkyl group, a CO-halogen substituted alkyl group, a CO-aryl group or a heteroaryl group; and wherein the compound of the formula VI has The following structure: 16 200831111

1 and 112 are independently hydrogen, alkyl, alkoxy, hydroxy, halogen; R3 and R4 are each independently hydrogen, a selectively substituted alkyl group, a branched 5-chain or a ring-based alkyl group, and optionally Substituted alkyl, branched or cyclic alcohols, optionally substituted alkyl chains, branched chains or cyclic amines. The methods available for the treatment of tumors and cancer are particularly affected by the toxic effects of guanidine, TCN-P and/or related compounds. In another embodiment, a method for treating a tumor in a mammal, particularly a human, comprising: (1) obtaining a biological sample from the tumor; (ii) determining whether the tumor overexpresses Akt kinase, and Iii) treating a tumor that overexpresses Akt kinase using a combination of triclinbine, tricineridine phosphate or a related compound and one or more onioncycline analogs as described herein. In another embodiment, the degree of Akt kinase expression can be determined, for example, by assaying the antibody in the acidified form to detect the 15 sputum acidified Akt kinase contained in the tumor or cancer. In another embodiment, the extent of Akt expression can be determined by examining the tumor or cancer cells obtained from the patient and comparing to the extent of the control tissue. In a particular embodiment, the Akt is at least 2, 2.5, 3 or 5 times overexpressed in the cancer sample as compared to the control tissue. In particular embodiments, the overexpressing Akt kinase 20 can be a highly active and phosphorylated Akt kinase. In another aspect of the invention, there is provided a dosing regimen that limits the toxic side effects of the associated compound of 2008 200811111. In another embodiment, such dosing regimens may minimize or remove toxic side effects including, but not limited to, hepatotoxicity, thrombocytopenia, hyperglycemia, diarrhea, gastritis And / or fever. In another embodiment, administration of TCN, 5 TCN-P or related compounds may result in at least a portion, such as at least 15, 20 or 30% or complete in vivo reaction, in at least 15, 20 or 25% of patients. In another embodiment, a method of treating a patient diagnosed with a tumor is provided according to a dosing schedule (which includes administering the troxaribine compound and the onioncyclin analog about once a week, which is time consuming An effective amount of TCN, TCN-P or related compound and one or more onioncycline is administered to the patient for about 3 weeks, followed by no administration of the triclinide compound and the onionine analog over a period of 10 weeks. analog. In another embodiment, there is provided a method of treating a tumor or cancer in a patient by administering to the patient a dosing regimen of 10 mg/m2 or less per week. TCN, TCN-P or phase 15 compounds and one or more anthracycline analogs. In another embodiment, the tricineridine compound and one or more onioncycline analogs can be administered in a single large dose over a short period of time, such as about 5, 10 or 15 minutes. In a further embodiment, there is provided a 20 dosing schedule wherein the tricineridine compound and one or more onioncyclins are administered by continuous infusion for at least 24, 28, 72, 96 or 120 hours. In a particular embodiment, the continuous administration can be repeated at least once a week, once every two weeks, and/or once a month. In other embodiments, the tricitin compound and one or more onioncycline analogs can be administered at least once every three weeks. In other embodiments, the compounds can be administered at least once a day for at least 2, 3, 4 or 5 days. 18 200831111 In a further embodiment, the patient can be administered the trozalbine compound and/or various onioncycline analogs as disclosed herein, in an amount effective to cause tumor regression. The administration of the triclinide compound and one or more onioncyclines may be at least partially obtained in at least 15 to 20% of the patient's tissues, such as at least 15, 20 or 3 %, or completely in vivo. . In a particular embodiment, the patient may be administered at least 2, 5, 1 〇, 15, 2 〇, 3 〇 or 5 〇 mg/m 2 of the triclinic compound disclosed in the text and one or more of the onioncyclines. Things. Administration of the triclinide compound and/or various onioncycline analogs can be carried out according to any of the treatment regimens disclosed herein. In a particular embodiment, the administration can include administration of less trimethoprim compounds less than 20 mg/m2 and one or more encycline analogs. In one embodiment, the trichostatin compound and one or more onioncycline analogs less than mg/m2 may be administered once a week. In other embodiments, the patient may be administered a dose of less than 2 mg/m 2, 5 mg/m 2, 10 mg/m 2, and/or 15 mg/m 2 of triclinbine compound and more than 15 types. Onioncycline analogue. In another embodiment, the patient may be dosed to less than 10 mg/m2 by continuous infusion for a period of at least 5 days. In a particular embodiment, the triclinide compound and one or more onionine analogs as disclosed herein can be used to treat pancreatic cancer, prostate cancer, colorectal cancer, and/or ovarian cancer. In another embodiment, the triclinide compound of the invention and one or more of the onioncycline analogs and/or treatment regimens can be used to prevent and/or treat cancer, sarcoma, lymphoma, leukemia, and/or Or myeloma. In other embodiments of the invention, the triclinide compound and one or more onioncycline analogs can be used to treat solid tumors. In still other embodiments, the tromethamine compound and one or more onioncycline analogs and compositions disclosed herein can be used in the treatment of a tumor or cancer, such as, but not limited to, the following organs or tissues: breast, Prostate, bone, lung, colon (including but not limited to colorectal), urinary system, bladder, non-Hodgkin's lymphoma, melanoma, kidney, adrenal gland, viscera, pharynx, thyroid, stomach, brain And/or ovaries. In 5, the triclinide compound and/or a plurality of cyclulin analogs can be used to treat pancreatic cancer, breast cancer, colorectal cancer, and/or ovarian cancer. In a further embodiment of the invention, the trichostatin compound and/or the onionine analog disclosed herein may be used to treat angiogenesis-related diseases. In a particular embodiment, a method of treating leukemia via continuous infusion of a triclinide compound and one or more anthracycline derivatives for at least 24, 48, 72 or 96 hours is provided. In other embodiments, continuous infusion may be repeated, such as at least once every 2, 3, or 4 weeks. In a specific implementation, a method for treating a tumor, cancer, and other disorders associated with abnormal cell proliferation in a host, the method comprising administering to the 15 host an effective amount of a pharmaceutically acceptable carrier The trichostatin compound and one or more onioncycline analogs. In the aspect of the invention, the triclinic compound and one or more onioncyclin analogs and compositions may be administered and may form part of the same composition, or may be adapted to be at the same time or 20 different compositions are available for administration at different times. In other embodiments, the tromethamine compound and one or more onionine analogs disclosed herein can be used to treat tumors or cancers that are resistant to - or a variety of known anticancer drugs, including tumors disclosed herein or Examples of cancers and compounds. In an embodiment, the amount of the trichostatin compound disclosed herein and 20 200831111 one or more onioncycline analogs are effective for treating a patient with a drug-resistant tumor or cancer, such as, for example, a drug-resistant tumor or cancer. Multidrug-resistant tumors or cancers, including, limited to: only anti-purple (four), anti-Raypa sin, 5 10 20 anti-tamoxifen, anti-shun #, and / or anti-gefitinib ( Gefltinib) (iressa) tumor or cancer. In a particular embodiment, a method is provided which comprises administering to a host in need of treatment an effective amount of a tripecline compound disclosed in the text and one or more onioncyclin derivatives, or a level thereof, effective to treat the host A pharmaceutical composition of a trichostatin compound and one or more onioncycline analogs of tumors, cancer, and other disorders associated with abnormal cell proliferation. In another embodiment, a method for treating a tumor or cancer is provided, which comprises a compound disclosed in the text, or a salt thereof, an isomer thereof, a prodrug 34, which is effective in the treatment of an individual. The cancer is, for example, a cancer, a sarcoma, a lymphoma, a leukemia or a myeloma. The compound or the tread, prodrug or s can be optionally selected to contain a suitable carrier (such as water); the formula = the composition of the pure supply, the complication of the compound is administered to the individual in need of treatment. The compound can be selectively treated with at least a therapeutic agent or in turn to treat a tumor or cancer. The scope of the present invention also includes a compound or a salt, prodrug or s thereof which is selectively disclosed in a pharmaceutically acceptable carrier, which is intended to treat a tumor or a dual use; and is optionally pharmaceutically acceptable. Revealing the use of a ciliben compound and/or a plurality of anthracycline analogs or salts thereof, for the preparation of a medicament for the treatment of cancer or a tumor.简单 Simple description of the diagram 21 200831111 Figure 1 illustrates the identification of API_2 (Querizine) as a suitable substance for the Akt inhibitor from the NCI Diversity Set. Figure 1A illustrates the chemical structure of API-2 (Quxi Libin). Figure 1B illustrates the phosphorylation sequence of API_2 inhibiting AKT2 in AKT2-transformed NIH3T3 cells. NIH3T3 cells transfected with wild-type AKT2 were treated with ΑΡΙ·2 (1 μΜ), time-consuming, and anti-phospho-Akt_T308 and -S473 antibodies (top group and intermediate group) were used for immunoblot analysis. The bottom group shows the performance of the total AKT2. In Figure 1C, it is shown that API-2 inhibits three variants of Akt. HEK293 cells were transfected with HA-Akt-AKT2 and -AKT3 10 and treated with either ΑΡΙ-2 (1 μΜ) or wortmannin (15 pM) before EGF stimulation, so that the cells were solubilized and anti-spasm Antibody immunoprecipitation. The immunoprecipitates were subjected to an in vitro kinase assay (top) and immunoblot analysis was performed using an anti-phospho-Akt-T308 (bottom) antibody. The middle group shows the expressiveness of the metastatically infected Aktl, AKT2 and AKT3. Figure 1D Figure 15 illustrates that API-2 does not inhibit Akt in vitro. An in vitro kinase assay of the recombinantly active AKT2 recombinant protein in a kinase buffer containing ΙμΜ API-2 (lane 3) was performed. Figure 2 illustrates that API-2 does not inhibit pi3K, PDK1, and closely related members of the AGC kinase family. Figure 2A illustrates an in vitro pI3K kinase assay. 20 Serum deficiencies of ΗΕΚ293 cells were treated with ΑΡΙ-2 (1 μΜ) or cleavage guanidine (15 μΜ) before E〇F stimulation, which took 3 minutes. The cells were lysed and immunized with an anti-ΡΙΙΟα antibody. The immunoprecipitates are subjected to an in vitro kinase assay using a matrix. The second picture shows the person ^. For the effect of in vitro activation of PDK1 (the top group), the filled circle indicates inhibition by 22 200831111 * 5 • API-2. The open circle indicates inhibition by the positive control staurosporine' which is a potent PDK1 inhibitor (iC50 = 5 nM). The bottom panel was an immunoblot analysis of HEK293 cells infected with Myc_PDK4 and treated with wortmannin or API-2 prior to EGF stimulation. The immunoblots are detected by the designated antibodies. Figure 2C illustrates the immunoblotting analysis of the degree of phosphorylation of PKCa using anti-phosphorylated PKCa T638 (top) and assembly PKCa (bottom) antibodies after treatment with ΑΠ_2 or non-selective PKC inhibitor R〇31 -8220. . Figure 2D shows the in vitro SGK kinase assay. HEK293 cells were transfected with HA-SGK and treated with API-2 or wortmannin prior to 10 EGF stimulation. In vitro kinase assays were performed using HAP as a substrate (top) with HA-SGK immunoprecipitates. The bottom panel represents the expressiveness of HA-SGK transfected with infection. Figure 2E illustrates the results of the PKA kinase assay. Immunopurified PKA was grown in ADB buffer (Upstate Biotechnology 15 Inc) containing the indicated inhibitor (API-2 or AKAI) and the substrate, Kemptide. The kinase activity was quantified. The 2F figure represents the Western dot method. Treatment of OVCAR3 cells with API-2 took time to specify. Immunoblots of cell lysates were performed with the indicated anti-phospho antibodies (Groups 1 to 4) and anti-actin antibodies (bottom). Figure 3 is a diagram showing that API-2 inhibits Akt activity and cell growth and induces apoptosis of human cancer cells with high Akt. Figure 3A shows Western blotting. After treatment with API-2, the phosphorylation of Akt was detected by anti-phospho-Akt-T308 antibody in designated human cancer cell lines. These dot methods were then probed with anti-total Akt antibodies (the bottom panel). Figure 3B shows a cell proliferation assay. The cell lines shown in the figure were treated with different doses of API-2 for a time of 24 23 200831111 and 48 hours, and then analyzed by the CellTiter 96 Cell Proliferation Assay Kit (Promega). Figure 3C provides a graph of cell death. The cells were treated with ΑΠ-2 and stained with annexin V and PI, and then analyzed by FACScan. Figure 4 shows that API-2 inhibits downstream targets of Akt and displays anti-tumor organisms in cancer cell lines with high Akt in mouse xenografts. Figure 4A shows that API-2 inhibits Akt acidification of potatorin, Bad, AFX and GSK-3/3. After treatment with API-2, OVAR3 cells were lysed and the ink spots were immunized with the indicated antibodies. Figure 4B shows that A PI-2 inhibits tumor growth. Tumor cells were injected subcutaneously into nude mice with low Akt cell content on the left and high Akt cell content on the right 10 sides. When the tumors reached an average size of about 100 to 150 mm 3 , the animals were treated with vehicle or 1 mg/kg/day of API-2. Each measured value represents the average of 10 tumors. Figure 4C is a graphical representation of mice treated with API-2 or vehicle (control) with VCAR3 (right) and OVCAR5 (left) xenografts. Figure 4D shows the tumor size (bottom) and weight (top) at the end of the experiment. In Figure 4E, anti-acidified Akt-S473 (top) and anti-AKT2 (bottom) antibodies were used in treated (T3 and T4) and untreated (T1 and T2) OVCAR-3-derived tumors. Immunoblot analysis of tumor lysates. Figure 5 shows that API-2 (Cucidine) inhibits in vitro kinase activity. In vitro kinase assays were performed with a recombinant form of P D K1 and A k t in a kinase buffer containing phospholipid creatinine-3,4,5-P3 (PIP3), API-2 and tissue protein as matrix. After 30 minutes of incubation, the aliquots were separated by SDS_PAGE and exposed to the film. Figures 6a to c provide the mRNA and amino acid sequence of human Aktl, also recorded in 2008 200811111. Figures 7a to d provide human Akt2 and mRNA amino acid sequences, as well as restriction enzyme positions. Figures 8a to c provide the mRNA and amino acid sequence of human Akt3, and also record the position of the restriction enzyme. c embodiment] detailed description of the preferred embodiment

In contrast to prior art and experience, the inventors have identified a method for successfully treating a combination of a triclinide compound and one or more cyclulin analogs for treating tumor 10 and cancer, by one or a combination of the following (1) Tritsulferine and/or a plurality of cyclulin analogs are administered only to patients who exhibit enhanced sensitivity according to the following diagnostic test for the triclinide compound and/or i-cyclin analog; (ii) use The dose which minimizes the toxicity of the tricineridine compound and/or the onioncycline analogs but which still has efficacy; or (d) the use of 15 to make the tricineridine compound and/or the like The dosage regimen wherein the toxicity of the cyclulin analog is minimized. DEFINITIONS As used herein, the term "compounds of the invention" is a salt of the compound VI and the salts thereof. 20 * The term is used in the text "cancer," and "cancerous," refers to or describes (4) the physiological condition of a whey animal with the characteristics of undeveloped cell growth (ie, augmentation disorder). (d) The real range of proliferative disorders, such as carcinomas, lymphomas, blastomas, sarcomas, and white ash; and other cancers described herein. More specific examples of such cancers include breast cancer, prostate cancer, knot 25 200831111 intestinal cancer, squamous cell carcinoma, small face, face n M, cell lung cancer, gastrointestinal cancer, neck cancer, ovarian cancer, liver cancer (eg Hepatocarcinoma), bladder cancer, endometrial cancer, kidney cancer, and thyroid cancer. * 5 10 15 20 Other non-limiting examples of cancer are basal cell carcinoma; biliary tract cancer; quaternary cancer; brain and CNS cancer; velvet larvae: osteochondral carcinoma; esophagus: cancer of the head and neck; Carcinoma; epithelial 赘m 艮, including Hodgkin's and non-Hodgkin's lymphoma; black phase; neuroblastoma; oral cancer (eg lip, tongue, mouth, and pharynx, sarcoma; Skin cancer; gastric cancer, Biwan II: = mouth, call the sputum cancer '· and other cancers and sarcomas. Sub-Lu cancer; urinary system cancer; as used in the text, the Lai, Xian all twins and hyperplasia (regardless of malignant $liang <|±), ,,,, cytogenesis " Chuansheng) and all precancerous and cancerous cell fish tissues. For example, a particular cancer may be characterized by a solid agglomeration. If present, when the solid tumor mass is present to the right, it is like a primary _ mass. The primary tumor mass ‘ is caused by the normal mass of the tissue | u. Mu a in the _ cancer cell material ί, can be by the existence of cysts (which can be found through visual or discovery) or by the shape of ribs, weight, structure or weight . However, some primary tumors are not tactile and can only be detected via medical imaging techniques, such as X-ray (e.g., breast radiography) or by needle aspiration. 'Second technology is more commonly used in early detection. Molecular and phenotypic analysis of cancer cells in a tissue can generally also determine whether the cancer is an endogenous cancer of the tissue or whether the lesion is due to metastasis from another site. 26 200831111

Unless otherwise specified, as used herein, the term "alkyl" includes, for example, <^ to (: 24 saturated linear, branched or cyclic, first, second or third hydrocarbons and more clearly married , including methyl, trifluoromethyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, pentyl, cyclopentyl, 5-isopentyl, Neopentyl, hexyl, isohexyl, cyclohexyl, cyclohexyldecyl, 3-methylpentyl, 2,2-dimercaptobutyl, and 2,3-dimethylbutyl. Substituting for, for example, one or more substituents such as halogen (F, C, Br or 1) (eg, CF3, 2·Βρ ethyl, CH2F, CH2Cb CH2CF3 or cf2cf3), hydroxyl (eg, ch2OH), amine (e.g., ch2nh2, 10 CH2NHCH3 or CH2N(CH3)2, an alkylamino group, an arylamino group, an alkoxy group, an aryloxy group, a nitro group, an azide group (e.g., CH2N3), a cyano group (e.g., ch2cn), a sulfonic acid, Sulfate, phosphonic acid, phosphate or phosphonate, and if necessary, as known to those skilled in the art' for example, as Greene et al. in Protective Groups ir> Omanic Synthesis, John Wiley and Sons 2 As taught in (1991), these groups may be unprotected or protected, and the document is incorporated herein by reference. Unless otherwise specified, the term "lower alkyl," Or a saturated alkyl group, a branched chain or, if appropriate, a cyclic alkyl group (e.g., a cyclopropyl group) which includes both substituted and unsubstituted forms. 20 The term "alkali-based, or,,, Aromatic amine groups, including amine groups having a - or a two alkyl or aryl substituent, respectively. The term "amino acid, including naturally occurring and synthetic alpha, stone or 5 amine groups & L and including' However, it is not limited to: amino acid found in protein (10), namely, glycine, alanine, lysine, leucine, isoleucine, and a broken 27 200831111 aminic acid, phenylalanine, tryptophan , valine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamic acid, aspartate, sulphate, lysine, essence Aminic acid, and histidine. In a preferred embodiment, the amino acid is in the L-configuration. Alternatively, the amino acid may be the following 5 derivatives: Sulfhydryl, amidino, leucine, iso-araminyl, amidino, phenylpropylamine, tryptamine, methionine, glycidinyl, amidino , sulphate, cysteamine, tyrosine, aspartame, glutamine, aspartame, lysine, spermine sulfhydryl, histamine sulfhydryl , /3-propylamine thiol, /3-amidoxime, leucine oxime 10, leucoylamine thiol, /3 yiisoguanamine sulfhydryl, phenyl-phenylpropylamine thiol, yS-color Amidoxime,/5-mercaptothioguanidino, /3-glycine thiol, S-seramine oxime, phenanthrenyl thiol, /3-cysteine thiol, /5-carbamide Sulfhydryl, S-aspartate amidoxime, /5-glutamamine oxime, aspartame, lysine, /3-spermine thiol, or /3 _ histamine sulfhydryl. When the term "amino acid" is used, its 15 is considered to be a specific and independent disclosure of each of the natural or synthetic amino acid esters in the D and L-configurations, including but not limited to: , Θ, r or glycine, alanine, valine, leucine, isoleucine, guanidine, phenylalanine, tryptophan, valine, serine, sulphamine Acid, cysteine, tyrosine, aspartic acid, glutamic acid, aspartate, glutamine 20 acid ester, lysine, arginine and histidine. The term "protected" as used herein, unless otherwise defined, includes a group that is intended to be added to an oxygen, nitrogen, sulfur or phosphorus atom to prevent its further reaction or for other uses. A variety of oxygen and nitrogen protecting groups are known to those skilled in the art of organic synthesis (see Greene and Wuts, Protective Groups in Organic 28 200831111).

Syntbesi 3rd Edition, John Wiley & Sons, Inc., New York, NYJ991) 〇

Unless otherwise specified, the term ''aryl, as used herein, includes phenyl, biphenyl or naphthyl' and is preferably phenyl. The aryl group may be optionally substituted by one or more molecular groups such as halogen, amine group, aromatine group, arylamine group, alkyl group, aryloxy group, nitro group, cyano group, and acid group. Sulfate, scaly acid, acid acid or phosphonate, and if necessary, as known to those skilled in the art, for example, in Greene et al., Protective Groups in Organic Synthpg^s John Wiley and Sons, 3rd Edition (1999). As taught, these molecular groups can be unprotected or protected. The term "alkaryl" or "alkyl", includes alkyl having an aryl substituent. The term "aralkyl" or "arylalkyl" includes aryl having an alkyl substituent. The term "halogen" as used herein includes chlorine, bromine, iodine, and oxygen. 15 The base includes citric acid' wherein the acetamino group is selected from a linear, branched or cyclic alkyl or lower alkyl group; an alkoxyalkyl group comprising a methoxymethyl group; An aralkyl group comprising a benzyl group; an aryloxyalkyl group, such as a phenoxymethyl group; an aryl group comprising a phenyl group optionally substituted by halogen, toxin (: 4 alkyl or (^ to alkoxy) Continuation of acid, such as alkyl or aryl sulfonyl, which includes methanesulfonyl, mono, di or triphosphate, trityl or monomethoxytriphenyl, substituted segments a trialkylcarbinyl group (e.g., dimethyl-tert-butylmethyl decyl) or a diphenylmethyl decyl group. The aryl group in the esters preferably contains a phenyl group. "Carbomethyl" means a fluorenyl group wherein the non-Weiyl group is a lower alkyl group. 29 200831111 As used herein, in terms of the purity of the mirror image, "substantially free, or lack of mussels," By means of a composition comprising at least 85 or 9% by weight, preferably from 9% to 5% by weight and more preferably from 99 to 8% by weight of the specified mirror isomer. In a preferred embodiment, The compounds and the compounds in the compounds are substantially free of other mirror image isomers. ▲ Similarly, the term "isolated" means at least 85 or 9% by weight, preferably 95 to 98% by weight and Still more preferably 99 to (10)% by weight of the compound and the compound composition containing other chemical species or mirror image isomers. 10 15 20 The term "independent" is used herein to mean the variable, which is applied independently. Independently different. Therefore, in a compound (such as R XYR'), 'where R' is "independently carbon or nitrogen, and both R" may be carbon, and two R" may be nitrogen 'or -R" may be Carbon and R- is nitrogen. The term "pharmaceutically acceptable salt or prodrug" is used throughout this specification to describe any pharmaceutically acceptable compound of the compound when administered to a patient. Acceptable forms (salts of (four), dish (iv), vinegar or related groups). Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic bases and acids. Suitable salts include those derived from the following salts. · Many of the pharmacy skills are well known to us Metals of other acids, such as unloading and sodium 'soil test metals' such as magnesium. Pharmaceutically acceptable prodrugs are compounds which can be metabolized, for example hydrolyzed or oxidized, in the host to form a compound of the invention. Typical examples include compounds having a biologically labile protecting group on a functional molecular group of the active compound. Prodrugs include oxidative, reductive, amination, deamination, hydroxylation, dehydroxylation, hydrolysis, dehydrolysis , burnt, de-sintering, brewing, deuteration, acidification, dephosphorylation to produce a compound of the active compound. 30 25 200831111 Unless otherwise specified, the term "pharmaceutically acceptable ester" as used herein is included Within the scope of normal medical judgment, it is applicable to one or more of the tissues that are in contact with the host and that are not toxic, irritating, allergic, etc., which are commensurate with reasonable benefit/risk ratio and can be effectively used for their purposes. The purpose of the five compounds. The term "patient" as used herein refers to an animal, preferably a mammal, preferably a human. Mammals may include non-human mammals including, but not limited to, pigs, goats, sheep, cattle (bovines), deer, lynx, horses, monkeys; and other non-human primates, such as dogs, Cat, rat, 10 mouse, rabbit or any other animal known or disclosed herein. Compounds of the Invention The present invention provides the use of TCN, TCN-P, a combination of a related compound and one or more onionine analogs, for the treatment of a particular therapeutic regimen for a proliferative disorder. 15 Unless otherwise specified, the term "tricilidine compound" and "tricilidine and related compounds" as used herein means a compound having the following structure:

31 200831111

Vk

Bazhonghan 2, R3' and R5' are each independently hydrogen, optionally substituted acid or phosphonate (including mono-, di- or tri-phosphate or stabilized phosphate prodrug); S4 a group (which includes a lower sulfhydryl group); an alkyl group (which includes a lower alkyl group); a decylamine, a sulphuric acid vinegar, which includes an alkyl group or an aralkyl group; a sulfonyl group including a methylsulfonyl group and a sulfhydryl group Wherein the phenyl group is optionally substituted with one or more substituents as described, for example, in the definition of an aryl group as taught herein; optionally substituted by 10; a lipid comprising a phospholipid; Amino acid; carbohydrate or cholesterol; or other pharmaceutically acceptable cleavage group in which R2, R3, and R5 are obtained in vivo as a compound of hydrazine or sulphur, -, " or tris-phosphate; Wherein JJX β 〇y and R are independently hydrogen, a selectively substituted phosphate; including low-stone anti-synthesis; _, a hospital base (which includes a low-carbon alkyl group); an aromatic salt oxidized olefin, such as Polyethylene glycol, optionally substituted aryl sulfhydryl, lipid, leucovorin, phospholipid; amino acid; carbohydrate Acceptable leaving group on a kill or other school;; a peptide; or cholesterol. In one embodiment, 32 15 200831111 the compound is administered as a 5,-phosphate ether lipid or s,_ether lipid.

Ri and R2 are each independently a linear, a branched or a cyclic alkyl group (which includes a lower alkyl group), an I α # 柯基基基基基基基基,CO-alkane a base, a CO-alkenyl group, a CO-alkynyl group, a CO-aryl group or a heterocyclic group, a CO group, a CO-alkoxyalkyl group, a CO-substituted aryl group, a CO-substituted aryl group, 碏酼苴^ * only sulfhydryl, alkanesulfonyl-arylsulfonyl, aromatic calcining base;

10 In the case of the shell, R2 and R3 are also chlorine. In another embodiment, R2, and R5 are hydrogen. In yet another embodiment, ν, R3, and R5 are hydrogen. In still another embodiment, r2', r3, R5, and RiAr^a. In another embodiment, the triclinic compound has the following structure: Ψ%

Wherein R 3 is a linear, optionally substituted straight chain, branched or cyclic alkyl group (which includes a low sulphonyl group), an alkenyl group or a blocked group, NH 2 , NHR 4 , N(R 4 ) 2 , 5 aryl, An oxo, aryloxy or substituted aryl; and each R4 is independently fluorene; fluorenyl, which includes a lower fluorenyl group; an alkyl group, including a low carbon alkyl group such as, but not limited to, methyl , ethyl, propyl and cyclopropyl; 33 200831111 alkenyl; alkynyl; cycloalkyl; alkoxy; alkoxyalkyl; hydroxyalkyl or aryl. In a sub-embodiment, it is a linear C1 to C11 alkyl, isopropyl, tert-butyl or phenyl group. In one embodiment, the kocillin compounds provided herein have a structure of 5 or less:

Has the following structure:

34 200831111 In another embodiment, the trichostatin compounds provided herein have the following structure:

5 wherein R6 is H, alkyl (which includes lower alkyl), alkenyl, alkynyl, alkoxyalkyl, hydroxyalkyl, aralkyl, cycloalkyl, NH2, NHR4, NR4R4, CF3, CH2OH, CH2F, CH2a, CH2CF3, c(y3)3, c(y3)2c(y3)3, C(=0)0H, C(=0)〇R4, c(=0)-alkyl, C(=0 - aryl, c(=0)-alkoxyalkyl, C(=0)NH2, C(=0)NHR4, C(=0)N(R4), wherein each 10 Y3 is independently H or halogen; And each R4 is independently H; fluorenyl, which includes a lower fluorenyl group; an alkyl group including a lower alkyl group such as, but not limited to, methyl, ethyl, propyl and cyclopropyl; alkenyl; alkyne a cycloalkyl group; an alkoxy group; an alkoxyalkyl group, a hydroxyalkyl group or an aryl group. In a sub-embodiment, 116 is ethyl, CH2CH2OH or CH2-styl. In another embodiment, the trichostatin compounds provided herein have the following structure: 35 200831111

Wherein R7 is hydrazine, halogen, alkyl (which includes lower alkyl), alkenyl, alkynyl, alkoxy, oxyalkyl, carbyl, cycloalkyl, decyl, cyano, 5 OH, OR4, NH2, NHR4, NR4R4, SH, SR4, CF3, CH2OH, CH2F, CH2a, 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 or N3, wherein each Y3 is independently H or halogen; and 10 each R4 is independently hydrazine; fluorenyl, which includes a lower fluorenyl group; an alkyl group, including a lower alkyl group, such as but not limited to Methyl, ethyl, propyl and cyclopropyl; alkenyl; alkynyl; cycloalkyl; alkoxy; alkoxyalkyl; hydroxyalkyl. In a sub-embodiment, R7 is methyl, ethyl, phenyl, chloro or hydrazine. In another embodiment, the Quercetin compound 15 provided herein has the following structure: 36 200831111

〇Ha

Formula XIII Formula XIV In one embodiment, the trichostatin compounds provided herein have the following structure:

37 200831111 The term "onionin analogue" as used herein, unless otherwise specified, refers to a compound of formula V or formula VI, wherein the compound of formula V has the structure:

The formula V 5 Ri, R 4 , the sigh 6 , and the R 7 are each independently a chlorine group, a base group, an alkoxy group; R 2 and each independently are hydrogen, an alkyl group, an alkoxy group, a hydroxyl group, a halogen; R 5 is a hydrogen, optionally substituted An alkyl chain, a branched or cyclic alkyl group, a trans group, an alkoxy group; R8 is a CO-alkyl group, a CO-halogen-substituted alkyl group, a CO-aryl group or a heteroaryl 10 group; and wherein the formula VI The compound has the following structure:

The formulae VI 1 and 112 are each independently hydrogen, alkyl, alkoxy, hydroxy, halogen; R 3 and R 4 are each independently hydrogen, optionally substituted alkyl chain, branched 15-chain or cyclic alkyl, optionally Substituted alkyl chain, branched chain or ring system 38 200831111 Alcohol, optionally substituted alkyl chain, branched chain or cyclic amine. In an illustrative embodiment, the term "formula v or formula VI onionine compound" encompasses, but is not limited to, compounds having any structure A to g:

5 A Idambidi B (Da deleted rubicin) 〇 small cranberries (D〇x〇nibicin)

D Epirubicin E Pimrubicin F Vairubicin

G Mitoxantrone 10 It should be understood that the compounds disclosed herein may contain a palm center. Such palm center may have a (R) or (S) configuration or may be a mixture thereof. Thus, the compounds provided herein may be mirror image-isomerically pure or may be a mixture of stereoisomerism or diastereomers. It is to be understood that the disclosure of the compounds herein encompasses racemic, optically active, polymorphic or stereoisomeric forms or mixtures thereof 39 200831111, preferably having the useful properties described herein, and methods for preparing optically active forms. Methods for determining activity using standard assays described herein or using other similar assays that are well known in the art are well known in the art. Examples of methods which can be used to obtain the optical isomers of the compounds include the following: Physical separation of crystals: A technique for manually separating macroscopic crystals of individual mirrored isomers. If the crystals of the individual mirror image isomers exist, that is, the substance is a crystal group and the crystals are visually different: simultaneous crystallization: only when the solution of the racemate is a solid crystal group, 10 can be respectively This solution crystallizes the technique of individual mirror image isomers. Enzymatic resolution: The technique of partially or completely separating the racemates by using enzymes at different rates of the reaction of the mirror image isomers. Enzymatic asymmetric synthesis: At least one step of the synthesis is the use of a catalytic reaction to obtain a synthesis technique of a pure or enriched hexa-form precursor of the mirror image isomer of the desired mirror image. Chemical asymmetric synthesis: a synthetic technique for synthesizing a desired image-isomer from a non-p-precursor precursor under conditions in which asymmetry (ie, palmarity) can be produced within the product. Catalytic or palmitic adjuvant; diastereomeric separation, reaction of the racemic compound with the mirror image isomer 20 pure reagent (the pair of palm adjuvant) to convert individual mirror image isomers A technique that is not a corresponding isomer. Chromatography or crystallization is then utilized to separate the diastereomers formed by their more pronounced structural differences, and the pair of palmitic adjuvants are later removed to obtain the desired mirror image isomers. First- and second-order asymmetry transformation: a diastereomer that balances the self-racemate 40 200831111 to the diastereomers from the desired mirror image isomer The preferential crystallization of the diastereomer in the solution or from the desired enantiomer of the mirror image will disturb the equilibrium so that in principle all of the material is converted from the desired mirror image to the crystalline diastereomeric Structure. Then 5 is the technique for releasing the desired mirror image isomer from the diastereomer. Kinetic Resolution Method · This technique refers to the partial or complete racemate of the racemic species under the kinetic conditions by the unique reaction rate of the mirror image isomers with palmitic, non-racemic reagents or catalysts. Resolution or further resolution of partially resolved compounds; 10 mirror-specific synthesis from non-racemic precursors: synthesis techniques for obtaining the desired mirror image isomers from non-preferential starting materials, and The stereochemical integrity of the synthesis process is not impaired or minimally impaired; for palm liquid chromatography: the separation of the racemate in the liquid mobile phase by the different interaction of the mirror image isomer and the stationary phase 15 techniques for these mirror image isomers. The stationary phase may be made of a palmitic substance or the mobile phase may contain another pair of palmar substances that can cause different interactions; for the palm gas layer method: volatilization of the racemate and borrowing of the mirror isomer in a gaseous state a technique for separating the mirror image isomers in a mobile phase with a different interaction with a column containing a fixed non-racemic palmitic absorber phase; 20 using a palmitic solvent extraction method, by means of a mirror image isomer A technique for preferentially dissolving the isomers in a specific pair of palm solvents; k is a technique for contacting a racemate with a film barrier by a method of carrying it on a military film. The barrier typically separates the two miscible fluids (one of which contains the racemate) and the driving force, such as concentration or pressure differential, can result in preferential transport by the film barrier via 41 200831111. In certain embodiments, a DMF adduct of triclinbine, tricineridine (TCN-P), trifluraline 5,-phosphate (TCN_P), or trifluraline is provided. (TCN-DMF). The TCN can be synthesized by any of the techniques known to those skilled in the art, for example, as described in Tetrahedron Letters, vol. 49, ρρ. 4757-4760 (1971). TCN-P can be prepared by any of the techniques known to those skilled in the art, for example, as described in U.S. Patent No. 4,123,524. The synthesis of TCN-DMF is described, for example, in inSERM, νο1·81, ρρ·37_82 (1978). Other compounds related to the TCN as described herein can be synthesized according to, for example, the method disclosed in the following document:

Gudmundsson, KS·, et al” “Synthesis of carbocyclic analogs of 2',3,-dideoxysangivamycin, 2',3,_dideoxytoyocamycin, and 25535-dideoxytriciribine/5 Nucleosides Nucleotides Nucleic, Acids, 20(10-11):1823- 1830 October-November 15 2001); Porcari, AR? et al.f a6-N-Acyltriciribine analogues: structure-activity relationship between acyl carbon chain length and activity against HIV-I/5 J. Med. Chem., 43(12 ): 2457-2463 (June 15? 2000); Porcari, AR? et aL, uAcyclic sugar analogs of triciribine: lack of antiviral and 20 antiproliferative activity correlate with low intracellular phosphorylatioa/9 Nucleosides Nucleotides, 18(11-12): 2475 -2497 (November-December 1999), Porcari, AR, et al. 9 6'Deoxy sugar analogues of triciribine: correlation of antiviral and antiproliferative activity with intracellular 42 200831111 phosphorylation," J. MM C7^m·, 43(12) :2438-2448(June 15, 2000),Porcari, AR,,a/" "Synthesis and antiviral activity of 2-substituted analogs of Triciribine," AW/eosiiies Nucleotides Nucleic Acids, 22(12): 2171-2193 (December 5 2003), Porcari, AR·, ei fl/, "An improved total synthesis of triciribine: a tricyclic nucleoside with antineoplastic and antiviral properties, " Nucleosides Nucleotides Nucleic, Acids, 23(1-2): 31-39 (2004), Schweinsberg, PD·, βία/.

"Identification of the metabolites of an antitumor tricyclic 10 nucleoside (NSC-154020) 5M Biochem. Pharmacol., 30(18): 2521-2526 (September 15? 1991)., Smith, KL? et aL, "Synthesis of new 2 '-beta-C-methyl related triciribine analogues as anti-HCV agents/9 Bioorg. Med. Chem. Lett., 14(13): 3517_3520 (July 5, 2004), Townsend, LB·, ei a/” “The 15 synthesis and biological activity of certain pentaazaacenaphthylenes, hexaazaacenaphthylenes and their corresponding nucleosidesZ, Nucleic Acids Symp. Ser.y 1986(17)··41-44 (1986), and/or Wotring, L, L·, with al·, “ Mechanism of activation of triciribine phosphate (TCN-P) as 20 a prodrug form of TCN/9 Cancer Treat Rep., 70(4): 491-7 (April 1986). pharmaceutically acceptable salts and prodrugs Where the nature or acidity is sufficient to form a stable non-toxic acid or base salt, the compound is preferably administered as a pharmaceutically acceptable salt. Pharmacy 43 200831111 The above acceptable salts include those derived from pharmaceutically acceptable inorganic or organic bases and acids Salt Included are alkali metals derived from many of the acids well known in the pharmaceutical art, such as potassium and sodium; and alkaline earth metals such as calcium and magnesium. In more detail, examples of pharmaceutically acceptable salts are those formed using acids. An organic 5 acid addition salt which forms a physiologically acceptable anion such as toluene, mesylate, acetate, citrate, malonate, tartrate, succinate, benzoate, ascorbate, ketone A glutaric acid, and an alpha-glycerophosphate. Also suitable are inorganic salts which include sulfates, nitrates, bicarbonates, and carbonates. ίο Standards that are well known in the art can be used. The procedure is to obtain a pharmaceutically acceptable salt, for example, by reacting a basic compound, such as an amine, with a suitable acid to give a physiologically acceptable anion. It can also be made into a test acid (such as sodium, potassium or a clock) or an alkaline earth metal (such as a mother). Any of the nucleotic acids described herein may be added with a nucleotate prodrug to increase the activity, bioavailability, stability, or properties of the nucleotide. Many nucleotide prodrugs are known for their coordination systems. In general, the alkylation, deuteration or other lipophilic modification of the mono-, di- or tri-structural acid salt of the nucleotide increases the stability of the nucleotate. Examples of substituents which may replace one or more hydrogens on the phosphate molecular group are alkyl groups, aryl groups, steroids, carbon 20 water compounds (which include sugars), 1,2-dimercaptoglycerols and alcohols. Many of these substituents are described in R· Jones and Ν· Bischofberger, Antiviral

Research, 27 (1995) 1-17. Either of these can be used in combination with the disclosed nucleoside acid to obtain the desired effect. In one embodiment, the triplicidin or related compound is provided as a 5'-trans base 44 200831111 lipophilic prodrug. Non-limiting examples of U.S. patents which disclose covalently incorporated into the nucleotide, preferably a suitable lipophilic substituent at the 5,_0H position of the nucleotide or lipophilic formulation, include U.S. Patent No. 5,丨49,794 (issued to % stealing et al. on September 22, 1992); 5,194,654 (issued to Hostetler et al., March 16, 1993); 55,522,263 (issued June 29, 1993) Give

Hostetler et al.; 5, 256, 641 (issued to Yatvin et al., October 26, 1993); 5, 411, 947 (to Hostetler et al., May 2, 1995); 5, 463, 092 (1 month, 1995) Presented to Hostetler et al. on the 31st; 5,543,389 (Yatvin et al., August 6, 1996); 5,543,390, 10 (issued to Yatvin et al. on August 6, 1996); 5,543,391 (1996) August 6 曰 to Yatvin et al.); and 5,554,728 (issued to Basava et al. on September 10, 1996), all of which are incorporated herein by reference. The disclosure of a lipophilic substituent or a lipophilic preparation which can be linked to the tromethine or related compound of the present invention includes W〇15 89/02733, WO 90/00555, WO 91/16920, W0 91/18914 WO 93/00910, WO 94/26273, WO/15132, EPO 350 287, EP 93917054.4, and WO 91/19721. A further non-limiting example of a derivative of tricineribine or a related compound is a derivative containing a substituent as described in the following publication. These derivatized trichostatin or related compounds can be used in the conditions described herein or as an antiviral agent, including anti-HIV or anti-HBV agents. The publications are as follows: Ho, DHW (1973) Distribution of Kinase and deaminase of Ιβ-D-arabinofuranosylcytosine in tissues of man and mouse. Cancer Res. 33? 2816-2820; Holy, A. (1993) Isopolar 45 200831111 phosphorous -modified nucleotide analogues. In: De Clercq (ed.), Advances in Antiviral Drug Design, Vol. I, JAI Press, pp. 179-231; Hong, CI·, Nechaev, A., and West, CR (1979a) Synthesis and antitumor activity of 1β-3-5 arabinofuranosylcytosine conjugates of cortisol and cortisone. Blochem. Biophys, Rs, Commun, 88, 1223-1229; Hong, CI5 Nechaev, A_, Kirisits, AJ Duchheit, DJ and West CR· ( 1980) Nucleoside conjugates as potential antitumor agents. 3. Synthesis and antitumor activity of 1-(β-ϋ-10 arabinofuranosyl) cytosine conjugates of corticosteriods and selected lipophilic alcohols. J, Med. Chem. 28? 171-177; Hostetler, KY ·, Stubmiller, LM·, Lenting, HBM van den Bosch, H. and Richman, DD (1990) Synthesis and antiretroviral activity of ph Ospholipid analogs of 15 azidothymidine and other antiviral nucleosides. J Biol Chem. 266, 11714-11717, Hostetler, Κ·Υ·, Korba, B. Sridhar, C., Gardener, M. (1994a) Antiviral activity of phosphatidyl-dideoxycytidine in Hepatitis B-infected cells and enhanced hepatic uptake in mice. Antiviral Res, 24? 59-67; Hostetler, 20 Κ·Υ·, Richman, DD, Sridbar, CN·Feigner, PL·, Feigerer, J·,

Ricci, J., Gerdencr, MF. Selleseth, DW· and Ellis, MN (1994b) Phosphatidylazidothymidine and phosphatidyl-ddC:Assessment of uptake in mouse lymphoid tissues and antiviral activities in human immunodeficiency 46 200831111 virus-infected cells and in rauscher leukemia virus -infected

Antimicrobial, Agents Chemother. 58, 2192-2191 \ Himston, RN·, Jones, AA· McGuigan, C·, Walker, RT·, Balzarini, J.? and De Clercq, E. (1984) Synthesis and 5 biological Properties of some cyclic phosphotriesters derived from 2,-deoxy-5-fluorouridine. J. Med. Chem. 27,440-444; Ji9 YH·, Moog, C·, Schmitt, G·, Bischoff, R and Luu, B· (1990 Monophosphoric acid diesters of 7P-hydroxycholesterol and of pyrimidine nucleosides as potential antitumor agents; 10 synthesis and preliminary evaluation of antitumor activity J. Med. Chem. 33? 2264-2270; Jones, AS, McGuigan, C.? waiter, RT ·, Balzarini, J. and DeClercq, E. (1984) Synthesis, properties, and biological activity of some nucleoside cyclic phosphoramidates. J. Chem. Soc. Perkin Trans. I? 1471-1474; 15 Juodka? BA and Smart, J (1974) Synthesis of ditribonucleoside a (PON) amino acid derivatives. Coll.

Czech. Chem, Comm. 39, 363-968; Kataoka, S., Imai, J., Yamaji, N., Kato, M., Saito, M., Kawada, T. and Imai, S. (1989) Alkylacted cAMP Dependent synthesis and 20 biological activities. Nucleic.Acids Res. Sym. Serf 21, 1-2; Kataoka, S·, Uchida, R. and Yamaji, N. (1991) A readily synthesis of adenosine 3', 5' Cyclic phosphate (cAMP) benzyl and methyl triesters, Heterocycles 32, 1351-1356; Kinchington, D., Harvey .J, J, O'Connor, TJ, Jones, 47 200831111 BCNM·, Devine, KG·, Taylor -Robinson, D., Jeffries, DJ and McGuigan, C. (1992) Comparison of antiviral effects of zidovudine phosphoramidate and phosphorodiamidate derivatives against HIV and ULV in vitro. Antiviral Chem. 5 Chemother· 3,107-112; Kodama, K· , Morozumi, M., Saitoh, Κ·Ι·, Kuninaka, H., Yoshino, H. and Saneyoshi, M. (1989) Antitumor activity and pharmacology of Ι-β-D-arabinofuranosylcytosine-55-stearylphosphate; an orally active Derivati Ve of Ι-β-D-arabinofuranosylcytosine. Jpn. J. 10 Cance Res. 80, 679-685; Korty, M· and Engels, J, (1979) The effects of adenosine- and guanosine 3”, 5'-phosphoric And acid benzyl esters on guinea-pig ventricular myocardium. Naunyn-Schmiedeberg's Arch· Pharmacol· 310, 103-111; Kumar, A·, Goe, RL., Jones, AS Walker, RT Balzarini, J. 15 and De Clercq, E (1990) Synthesis and biological evaluation of some cyclic phosphoramidate nucleoside derivatives. J. Med, Chem. 335 2368-2375; LeBec, C.? and Huynh-dinh, T. (1991) Synthesis of lipophilic phosphate triester derivatives of 5- 20 fluorouridine and arabinocytidine as anticancer prodrugs. 20 Tetrahedron Lett. 32, 6553-6556; Lichtenstein, J. Barner, HD and Cohen SS (1960) The metabolism of exogenously supplied nucleotides by Escherichia coli, J Biol. Chem. 235, 457- 465; Lucthy, J., Von Dacniken, A., Friederich, J.

Manthey, B., Zweifel, J., Schlatter, C. and Benn, Μ·Η· (1981) 48 200831111

Synthesis and toxicological properties of three naturally occurring cyanoepithioalkanes. Mitt. Geg. Lensmittelunters. Hyg. 12, 131-133 (Chem. Ahstn 95? 127093); McGuigan, C. Tollerfield, SM and Riley, PA (1989) Synthesis and 5 biological Evaluation of some phosphate triester derivatives of the anti-viral drug Ara. Nucleic, Acids Res. 17? 6065-6075; McGuigan, C·, Devine, KG, O'Connor, TJ, Galpin, SA·, Jeffries, DJ and Kinchington D. (1990a) Synthesis and evaluation of some novel phosphoramidate derivatives of 10 3,-azido-3 '-deoxythymidine (AZT) as anti-HIV compounds.

Antiviral Chem. Chemother 15 107-113; McGuigan, C.? O'Connor, TJ·, Nicholls, SR Nickson, C. and Kinchington, D. (1990b) Synthesis and anti-I-HIV activity of some novel substituted dialkyl phosphate Derivatives of AZT and ddCyd. 15 Antiviral Chem. Chemother 1? 355-360; McGuigan, C.9

Nicholls, SR·, O'Connor, TJ·, and Kinchington, D. (1990c) Synthesis of some novel dialkyl phosphate derivative of 3'-modified nucleosides as potential anti-AIDS drugs. Antiviral Chem. Chemother 1, 25-33; McGuigan, C.9 Devine, 20 KG, O'Connor, TJ·, and Kinchington, D· (1991) Synthesis and anti-HIV activity of some haloalky phosphoramidate derivatives of 3, azido-3'deoxythylmidine (AZT); potent activity Of the trichloroethyl methoxyalaninyl compound. Antiviral Res. 15? 255-263; McGuigan, C.? Pathirana, RN, 49 200831111

Mahmood, N” Devine, KG and Hay, AJ· (I 992) Aryl phosphate derivatives of AZT retain activity against HIVI in cell lines which are resistant to the action of AZT. Antiviral and Lie 17, 311-321; McGuigan, C · Pathirana, R, N·, Choi, SM·, 5 Kinchington, D· and O'Connor, TJ (1993a)

Phosphoramidate derivatives of AZT as inhibitors of HIV, studies on the caroxyl terminus. Antiviral Chem. Chemother 4, 97-101; McGuigan, C., Pathirana, RN·, Balzarini, J. and De Clercq, E. (1993b) Intracellular delivery The bioactive AZT 10 nucleolides by aryl phosphate derivatives of AZT. J Med Chem. 36, 1048-1052. The anti-HIV agent AZT hydrogen hydride alkylate derivative is less toxic than the parent nucleotide analog. Heart r·· 5, 271-277; Meyer, RB·, Jr., Shuman, DA· and Robins, RK (1973) 15 Synthesis of purine nucleoside 3', 5'-cyclic phosphoramidates· Tetrahearon Lett. 269-272; Nagyvary, J. Gohil, RN,

Kirchner, CR and Stevens, JD (1973) Studies on neutral esters of cyclic AMP? Biochern. Biophys. Res. Commun. 55? 1072-1077; Namane, A. Goyette, C·, Fillion, MP·, Fillion, G· 20 and Huynh-Dinh? T. (1992) Improved brain delivery of AZT using a glycosyl phosphotriester prodrug. J. Med. Chem. 35, 3939-3044; Nargeot J. Nerbonne, JM Engels, J. and Leser, HA (1983) Natl. Acad. Sci. USA 80, 2395-2399; Nelson, KA, Bentrude, WG, Stser, WN and Hutchinson, JR (1987) 50 200831111

The question of chairtwist equilibria for the phosphate rings of nucleoside cyclic 39,59-monophosphates. ^NMR and x-ray crystallographic study of the diasteromers of thymidine phenyl cyclic 39,59-monophosphate. J. Am. Chem. Soc. 109? 4058-4064; Nerbonne, JM” Richard, S., Nargeot J. and

Lester, H.A. (1984) New photoactivatable cyclic nucleotides produce intracellular jumps in cyclic AMP and cyclic GMP concentrations. Nature 301, 74-76; Neumann, J.M., Herve,

M·, Debouzy, JC·, Guerra, FI·, Gouyette, C., Dupraz, B. and 10 Huynh-Dinh, T (1989) Synthesis and transmembrane transport studies by NMR of a glucosyl phospholipid of thymidine· J· Am· Chem·Soc· 111,4270-4277; Ohno, R·, Tatsumi, N·, Hirano, M·, Imai, K. Mizoguchi, H·, Nakamura, T·, Kosaka, M·, Takatuski, K., Yamaya , T·, Toyarna, K·, 15 Yoshida, T·, Masaoka, T·, Hashimoto, S·, Ohshima, T·,

Kimura, I.9 Yarnada, K. and Kimura, J. (1991) Treatment of myelodyspastic syndromes with orally administered 1 -P-D-rabinofuranosylcytosine-59 -stearylphosphate.

Oncology 48, 451-455· Palomino, E., Kessle, D. and Horwitz, 20 J?P. (1989) A dihydropyridine carrier system for sustained delivery of 2,?3,dideoxynucleosides to the brain. J. Med, Chem 32, 622-625; Perkins, RM·, Bamey, S., Wittrock, R., Clark, PH·, Levin, R. Lambert, DM, Petteway, S, R., Serafinowska, HT·, Bailey, SM ·, Jackson, S., Harnden, MR·, 51 200831111

Ashton, R., Sutton, D., Harvey, JJ and Brown, AG (1993) Activity of BRL47923 and its oral prodrug, SB203657A against a rauscher murine leukemia virus infection in mice. Antiviral Res. 20 (Suppl. I). 84 Piantadosi, C" Marasco, CJ" 5 Jr·, Morris-Natscbke, SL·, Meyer, KL·, Gumus, F·, Surles, JR·, Ishaq, KS·, Kucera, LS lyer, N·, Wallen, CA·, Piantadosi, S. and Modest, EJ (1991) Synthesis and evaluation of novel ether lipid nucleoside conjugates for anti-HIV-1 activity. J. Med. Chem. 34? 1408-1414; Pompon, 10 A·, Lefebvre , I·, Imbach, JL·, Kahn, S. and Farquhar, D. (1994) Decomposition pathways of the mono- and bis (pivaloyloxyrnethyl) esters of azidothymidine-5,-monophosphate in cell extract and in tissue culture medium; Application of the on-line ISRP-cleaning9 HPLC technique. 15 Antiviral Chem. Chemother. 5, 91-98; Postemark, T. (1974) Cyclic AMP and cyclic GMP. Anu. Rev. Pharmacol. 14, 23-33; Prisbe ,EJ·,Martin,J .CM·, McGee, DPC·, Barker, MF·, Smee, DF Duke, AE·, Matthews, TR and Verheyden, JPJ (1986) Synthesis and antiherpes virus activity of phosphate 20 and phosphonate derivatives of 9-[(l? 3-dihydroxy-2-propoxy) methyl] guanine· J. Med, Chem· 29, 671-675; Pucch, F·, Gosselin, G, Lefebvre, I, Pompon, A·, Aubertin, AM· Dirn, A And Imbach, JL (1993) Intracellular delivery of nucleoside monophosphate through a reductase-mediated 52 200831111

Antiviral Res. 225 155-174; Pugaeva, VP? Kochkeva, SI5 Mashbits, FD and Eizengart, RS (1969). Toxicological assessment and health standard ratings for ethylene sulfide in the industrial atmosphere. Gig. Trf Prof 5 Zabol 13, 47-48 (Chem. Abstr. 72? 212); Robins, RK (1984) The potential of nucleotide analogs as inhibitors of retroviruses and tumors. Pbarm. Res. 11-18; Rosowsky, A.? Kim5 SH? Ross and J. Wick, MM (1982) Lipophilic 55-(alkylphosphate) esters of Ι-β-D-arabinofuranosylcytosine 10 and its TV^-acyl and 2.25-anhydro-3O-acyl derivatives as potential prodrugs· J· Med,Chem · 25, 171-178; Ross, W· (1961) Increased sensitivity of the walker turnout towards aromatic nitrogen mustards carrying basic side proteins following glucose pretreatment. Biochem. Pharm. 8? 235-240; 15 Ryu, EK·, Ross, RJ·, Matsushita, T., MacCoss, M., Hong, CI and West, CR (1982). Phospholipid-nucleoside conjugates 3. Synthesis and preliminary biologica l evaluation ofl-PD-arabinofuranosylcytosine5'diphosphate[-],2-diacylglycerols. J.Med.Chem. 25,1322-329; Safthill, R. and 20 Hume, WJ (1986) The degradation of 5-iododeoxyurindine and 5- Bromoeoxyuridine by serum from different sources and its consequences for the use of these compounds for incorporation into DNA. Chem. Biol. Interact. 57, 347-355; Saneyoshi, M., Morozumi, M., Kodama, K., Machida, J ·, 53 200831111

Kuninaka, A. and Yoshino, H. (1980) Synthesis and biological evaluations of a series of Ι-β-D-arabinofuranosylcytosine 55-alkyl or arylphosphates. Chem5 Pharm. Bull. 28? 2915-2923; Sastry, 5 JK·, Nehete, PN·, Khan, S., Nowak, BJ., Plunkett, W_,

Arlinghaus, RB and Farquhar, D. (1992) Merabrane-permeable dideoxyuridine 5'-monophosphate analogue inhibits human immunodeficiency virus infection. Mol. Pharmacol, 41, 441-445; Shaw, JP, Jones, RJArimilli, 10 M, N· , Louie, MS,, Lee, WA and Cundy, KC (1994) Oral bioavailability of PMEA from PMEA prodrugs in male Sprague-Dawley rats. 9th Annual AAPS Meeting. San Diego, CA (Abstract)· Shuto, S·, Ueda , S., Imamura, S., Fukukuawa, K. Matsuda, A. and Ueda5 T. (1987) A facile one-step 15 synthesis of S ^ phosphatidyl nucleosides by an enzymatic two-phase reaction. Tetrahedron Lett. 28? 199- 202; Shuto, S.? Itoh,.H., Ueda, S., Imamura, S., Kukukawa, K., Tsujino, M. Matsuda, A. and Ueda, T. (1988) A facile enzyatic synthesis of 5 ,-(3-sn-phosphatidyl) nucleosides and their antileukemic 20 activities. Chem. Pharm· Bull· 36, 209-217. A preferred orthoester prodrug group is S-mercapto-2-thioethyl, which is also known as "SATE". Additional examples of prior drugs may be used 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 54 200831111

No. 6,252,060, 6,448,392, 5,411,947, 5,744,592, 5,484,809, 5,827,831, 5,696,277, 0,022,029, 5,780,617, 5,194,654, 5,463,092, No. 5, 744, 461, 4, 444, 766, 4, 562, 179, 5, 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, 6,670,341, 2004/0161398, 2002/082242, 5,512,671, 10 2002/0082242, and/or PCT Publication No. WO 90/11079, WO 96/39197' and/or WO 93/08807. In vivo efficacy/dosing regimen In another aspect of the invention, there is provided a dosing regimen that limits the toxic side effects of TCN and related compounds. In one embodiment, such administrations may minimize toxic side effects including, but not limited to, hepatotoxicity, thrombocytopenia, hyperglycemia, vomiting, hypo-5emia , anemia, hypoalbuminemia, inhibition of gastric myeloid activity, hypertriglyceridemia, high amylase whiteness, diarrhea, osteitis and/or fever. In another embodiment, administration of TCN, TCN-P or related compounds and one or more than 20 onioncycline analogs may result in at least partial or complete in vivo response in at least 15 to 20% of patients, in a particular implementation In one embodiment, a portion of the 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 present invention can be found in at least 15, 20, 25, 30, 35, 40, 50, 55, 60, 65, 70, 75, 55 200831111 80, 85, or 90% of the treated patients. reaction. In other embodiments, such response rates can be obtained by any of the treatment regimens disclosed herein. In its mussel case, a method for treating a disease diagnosed as having cancer is provided by administering an effective amount of TCN, TCN_P or related human private 7 u ^ according to a dosing schedule. σ and one or a mixture of onioncycline analogs, the dosing schedule comprising administering the trichostatin compound and/or the onioncycline analog once a week for 3 weeks, followed by no vote for a week The drug is administered (ie via a 28 day cycle). In other embodiments, such 28-day week 10 h 2, 3, 4, or 5 times may be repeated or until the tumor is clearly found to subside. In another embodiment, a 42-day cycle is provided, the sulphate disclosed in the Chinese may be administered once per week, taking 4 weeks, and then the tromethamine is not administered during 2 weeks. : € cyclin analogues. In other embodiments, such a 42 day cycle can be repeated for at least 2, 3, 4 or 5 times or until the apparent 15 is found to subside. In a particular embodiment, less than 12, less than η, or less than 1 〇 mg/m 2 of D, TCN_p or related compounds are added. In other specific embodiments, 2, 3, 4, 5, 6'7, 8'9, 1 or 11 mg/m2 may be added according to the day of the day (:.1(::1) ^1> or a related compound. In another specific embodiment, about 1 to about 50 gram of cyclidine derivative is administered. In a particular embodiment, it can be administered according to a periodicity. 15, 15, 20, 25, 30, 35, 40, 50 or 100 mg of cyclcyclin derivatives. In the case of "Beibei &", it provides a method for treating cancer in a patient, which is administered to the patient once a week. Dosing regimen of 1 mg/m 2 or less of TCN, TCN_p 56 200831111 or related compounds and less than about 30 mg of one or more onioncycline analogs. In a particular embodiment, it may be administered once a week. 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4, 5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 mg/m 2 as disclosed herein TCN, TCN-P or related compound. In another specific embodiment, 1, 5, 10, . 15, 20, 25, 30, 35, 40, 50 or 100 mg may be administered once a week. An onion ring derivative. In an embodiment of the invention, In a short period of time, for example about 5, 10, 15, 20, 30 or 60 minutes. The gastric compounds disclosed herein are administered simultaneously in a single large dose. In further embodiments, there are provided in which the compounds are infused by 10 consecutive infusions. a dosing schedule that takes at least 24, 48, 72, 96, or 120 hours to be administered at the same time. In a particular embodiment, the tricineridine compound can be administered by repeated or bulk injections at a specific frequency as described below. / or onionin analogues: at least once a week, at least once every two weeks, at least once every three weeks, at least once a month, at least once every five weeks, at least once every six weeks, at least once every six weeks At least once every 10 weeks and/or at least once every 12 weeks. ^ The type and frequency of administration can be combined to create a dosing cycle using any of the methods disclosed herein. The Quxi can be repeatedly administered by a specific dosing cycle. The compound and/or the onionine analog, for example, is administered in a large injection every two weeks, and takes 3 weeks. The administration period can be administered, and the time is at least: 1, 2, 20 3, 4 , 5, 6, 7, 8, 9, 10, 1 1, 12, 18 or 24 months. Alternatively, the patient may be administered at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15 or 20 dosing cycles. The tricitin compound and/or the onioncycline analog is administered in any combination disclosed, for example, the triclinide compound and/or the onioncycline analog can be administered once a week, once every 3 weeks. 3 cycles. 57 200831111 In further embodiments, the compounds may be administered separately once a day for at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 days. After such administration, the triclinide compound and/or the onioncycline analog was not administered during the same period of time. 5 The patient may be administered TCN, TCN-P and related compounds as disclosed herein with one or more anthracycline analogs, the amount of which is effective to cause tumor regression. Administration of the TCN, TCN-P or related compound and one or more anthracycline analogs may result in at least a portion, such as at least 15, 20 or 30%, or at least in vivo in at least 15 to 20% of such patients. reaction. In certain embodiments, the patient may be administered at least 2, 5, 1 〇, 15, 2 〇, 25, 3 〇 or 503 ⁄4 g/m 2 of the triclinide compound disclosed in the text. In a particular embodiment, the patient may be administered 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, 12, 15, 17, 20, 25, 3〇, 35, 40, 45, 50, 55, 60 '65, 70, 15 75, 8〇, 85, 90, 95, 100 TCN, TCN-P or related compounds disclosed in the text 150, 165, 175, 200, 250, 300 or 35 mg/m 2 can be administered to patients in specific embodiments, 5, ^, 2 〇, 25, 3〇, 35, 4G, 5G or 1GG mg of onioncycline derivatives. Administration of the compound is carried out according to the treatment regimen disclosed herein. In the case of the special scorpion, the dosing regimen comprises simultaneously, continuously or over a period of time at about 2 oz/m 2 of TCN and related compounds with less than about 3 〇 克 克 克 素. In the embodiment, the hormone can be administered at the same time once a week. In another embodiment, less than 2〇58 200831111 mg/m2 of TCN or related compound may be administered weekly and less than about 3 mg of onioncycline derivative may be administered in the next week. In further embodiments, the patient may be administered 2 mg/m 2, 5 mg/m 2, 10 mg/m 2, and/or 15 mg/m 2 of TCN4-related compound and less than about 5 30, 25, 20 , 15 or 10 mg of onioncycline derivatives. In another embodiment, the patient can be administered less than 10 mg/m2 of tripellidine compound and less than about 30 mg of onioncyclin derivative via continuous infusion for a period of at least 5 days. The invention provides

Any combination of the type of administration, frequency, number of cycles, and dosage disclosed in the text. In addition to the patient population, in one aspect of the invention, methods are provided for identifying cancers or tumors that are susceptible to the toxic effects of Tricinem (TCN) and related compounds. In one embodiment, a method of treating a cancer or tumor in a mammal by (1) obtaining a biological sample from the tumor; (9) determining whether the cancer or tumor overexpresses Akt kinase or a highly activated and acidified tyrosine kinase And (iv) treating the cancer or tumor with the use of triclinide or a related compound as described in the text. In one embodiment, the biological sample can be a biopsy. In other embodiments, the biological sample can be a fluid derived from the tumor or cancer, and the biological sample can be obtained according to any technique known to those skilled in the art. In the implementation, a biopsy can be performed to obtain the biological sample. Procedures for biopsy to remove tissue or cells from the body for testing Some biopsies can be performed in the physician's office, while other biopsies: To be smashed in::1⁄4. In addition, some biopsies require the use of anesthesia for the hemp phase area while other biopsies do not require any sedation for use. In a particular embodiment, endoscopic biopsy can be performed. Such biopsies are performed through a natural body hole (i.e., a rectum) or a small incision (i.e., arthroscopy) via a light mirror (a long, thin tube suitable for viewing in a lens having the closest focus). The endoscope is used to view an abnormality 5 or suspicious area of the organ to obtain a small amount of tissue suitable for the study. Endoscopy procedures are used for organs or body areas that are to be developed and/or treated. The physician can insert the endoscope into the gastrointestinal tract (endoscopic gastrointestinal examination), bladder (cytoscopic examination), abdominal cavity (laparoscopic examination), joint cavity (joint examination), middle part of the chest (longitudinal examination) or Within the gas officer and bronchial system (laryngeoscopy and bronchoscopy). In another embodiment, bone marrow biopsies can be performed. Such biopsies can be performed from the sternum or iliac spine (the area of the bone on either side of the pelvis on the lower back area). Wash the skin and provide a local anesthetic to paralyze the area. Insert a long hard needle into the marrow and aspirate the cells for study; this step is occasionally uncomfortable. After the aspiration step, a core living body 15 slice can be taken (from the pulp to remove the small bone "fragment,"). In another embodiment, the mammal can be subjected to a resectable or incisional biopsy. When more width is required Or a deeper skin area, usually this biopsy. Use a scalpel (scalpel) to remove the full thickness of the skin for further examination, and suture (using surgical sutures) the wound. When removing the entire tumor 20 When it is called a resectable biopsy technique, if only one part of the tumor is removed, it is called a biopsy technique. For example, when a melanoma (a kind of skin cancer) is suspected, the resected living body Sectioning is usually a method that is often preferred. In still other embodiments, fine needle aspiration (FNA) biopsies can be used. 2〇〇83liii. 5 • This method of biopsy involves the use of fine needles to move from the tumor. Except for small tablets. Sometimes local anesthetics are used to itch this area, but this test rarely causes discomfort and leaves no symptoms. FNA is not suitable, such as the diagnosis of suspected pigmented nevus, but can be used, for example, live Check for large lymph nodes close to melanoma to see if the melanoma has metastasized (diffusion). A computed tomography scan (CT or CAT scan) can be used to introduce the needle into a tumor in an internal organ such as the lung or liver. In other embodiments, drilling, shaving, and/or biopsy of the skin may be performed. The drill biopsy includes a deep biopsy instrument using a short cylinder of removable tissue or a 'small core core'. Skin sample. After administration of the topical anesthetic, the instrument is rotated on the surface of the skin until it passes through all layers (including the dermis, the epidermis, and the most surface portion of the subcutaneous tissue (fat)). A shaved living section includes removing the layer by shaving off the uppermost layer of the skin. A local anesthetic is also used for shaving biopsy. Skin biopsy 15 • Tablets include removal of skin samples for examination under a microscope to determine, for example, the presence of melanoma. This biopsy method is performed under local anesthesia. In a particular embodiment, a method is provided to determine if the tumor overexpresses Akt kinase. Overexpression of Akt kinase may indicate the phosphorylation status of the kinase. The high phosphorylation of Akt can be detected according to the methods described herein. In one embodiment, tumor biopsies and control tissues can be compared. The control tissue may be a normal tissue obtained from a mammal obtaining the biopsies or a normal tissue obtained from a healthy mammal. Akt kinase overexpression or high acid nucleation can be determined to understand whether the content of the tumor biopsy and/or Akt kinase phosphorylation is higher than the control tissue, such as Akt 61 200831111 kinase content than the control tissue The Akt kinase content is at least about i5, 2, 2.25, 2.5, 2.75, 3, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.5, 6, 7, 8, 9, or 10 times. In one embodiment, the invention provides a method of detecting abnormal Akt kinase expression in a patient or a biological sample obtained from the patient, by using a specificity for Akt kinase or an antigenic portion thereof The immunointeractive molecule contacts cells, cell extracts, serum or other samples or biological samples obtained from such patients and screens for immunocompetent molecule-Akt kinase complex formation, wherein the relative cellular cells The high presence of the complex indicates abnormal cells that can express or overexpress Akt. In one embodiment, the amount of high Akt kinase present in cells or cell extracts can be screened immunologically. In another embodiment, the degree of expression of the gene encoded by Akt kinase can be screened to detect abnormal expression of Akt in the cell at the gene level, wherein the transcriptional expression product (ie, mRNA) is compared with normal cells. A high content of 15 indicates abnormal cells. In particular embodiments, real-time PCR as well as other PCR programs can be used to determine transcriptional activity. In one embodiment, the mRNA can be obtained from a patient's cell or a biological sample obtained from the patient and can selectively produce cDNA. The mRNA or cDNA can then be contacted with a gene probe capable of hybridizing and/or amplifying all or part of the nucleotide sequence encoding the Akt kinase-encoding nucleotide sequence or its complementary nucleotide 20, and then detecting the mRNA or cDNA content. , wherein the presence of high levels of mRNA or cDNA compared to the normal control group can be assessed. Still another embodiment of the present invention contemplates the use of antibodies (single or multiple strains) of Akt kinase in a quantitative or semi-quantitative diagnostic kit to determine the relative amount of Akt kinase in a patient's suspected cancer cell 62 200831111 * 5 cells, The kit can include all of the reagents required to perform the assay. In one embodiment, a kit for using the reagents and materials required for performing an ELISA assay is provided. The reagent may include, for example, a buffer for washing, an antibody dilution buffer, a buffer for blocking, a cell staining solution, a developing solution, a stop solution, an anti-phosphoprotein specific antibody, an anti-Pan protein-specific antibody, a secondary antibody. And distilled water. The kit may also include instructions for use and may be selectively automated or semi-automated or in a form that is compatible with automated machinery or software. In one embodiment, a phospho-serine 10 473Akt antibody capable of detecting an activated form of AKT (phosphorylated Akt of serine 474) can be used as an antibody in a diagnostic kit. See, for example, Yuan et al. (2000) uFrequent Activation of AKT2 and induction of apoptosis by inhibition of phosphinositide-3-OH kinase/Art pathway in human ovarian cancer, '' Oncogene 19:2324-2330 ° Akt kinase 15 • Akt, also Known as PKB3, it represents a subfamily of serine/threonine kinases. Three members have been identified in this subfamily, AKT1, AKT2, and AKT3. In the PI3K-dependent manner, Akt is activated by extracellular stimuli (Datta, S. R., et al. Genes Dev. / 3: 2905-2927, 1999). Complete activation of Akt requires phosphorylation of Thr3〇8 in the activation loop and phosphorylation of Ser473 in the C-terminally active domain. Akt is poorly regulated by PTEN tumor suppressor. Mutations within PTEN have been identified in various tumors that can lead to activation of the Akt pathway (Datta, S. R., et al. Genes Dev. 73: 2905-2927, 1999). In addition, amplification, overexpression and/or activation of Akt has been detected in many human malignancies (Datta, SR, et al. Genes Dev. 63 200831111 73: 2905-2927, 1999, Cheng, J. Q., And Nicosia, S. V. AKT signal transduction pathway in oncogenesis. In Schwab D, editor. Encyclopedic Reference of Cancer. Berlin Heidelberg and New York: Springer; 2001. pp35-7). Akt, especially for the ectopic expression of the orthologous 5-akt, induces cell survival and malignant transformation, whereas inhibition of Akt activity stimulates apoptosis in a range of mammalian cells (Datta, SR, et al. Dev. 73:2905-2927, 1999, Cheng, J. Q., and Nicosia, S. V. AKT signal transduction pathway in oncogenesis. In Schwab D? editor. Encyclopedic Reference of 10 Cancer. Berlin Heidelberg and New York: Springer 2001. pp35-7, Sun, M., et al. Am. J. Path., 159: 431-437, 2001, Cheng, JQ" et al. 〇nc〇gene, 14: 2793-2801, 1997). Activation of Akt has been shown to be associated with tumor invasiveness and chemical resistance (West, KA·, et al. Dmg Resist. Updat., 5: 234-248, 2002). 15 Activation of the Akt pathway in malignant transformation and Chemical resistance plays a key role in inducing cell remnant, growth, migration, and angiogenesis. The present invention provides methods for determining the overexpression and/or high activation of Akt kinase and the content of alkalylated Akt kinase. Akt kinase can be any known Akt kinase or Related kinases, 20 include, but are not limited to, Aktl, Akt2, Akt3. The mRNAs of human Aktl, Akt2, Akt3 and amino acid sequences are elucidated in the maps of ruthenium, 7a_d, and 8a-c, respectively. Immunological Assay 64 200831111 In one embodiment, a method for detecting abnormal expression of Akt kinase in a mammalian cell or in a biological sample obtained from the mammal is provided by using Akt kinase Or an immunologically interacting molecule having specificity in contact with the mammalian cells, cell extract 5 or serum or other sample or biological sample, and screening for immunocompetent molecule-Akt kinase complex The extent of the formation of the substance, and then whether the complex is present in a large amount relative to the normal cells. The immunointeractive molecule may be specific to and bind to Akt kinase or an antigenic portion thereof or a homolog thereof or a derivative thereof. Affinity is divided into 10. In one embodiment, the immunointeractive molecule can be an immunoglobulin molecule. In other embodiments, the immunoglobulin molecule can Antibody fragments, single chain antibodies, and / or deimmunized molecules including humanized antibodies and T-cell associated antigen-binding molecules (TABM). In a specific embodiment, the antibody can be a monoclonal antibody. In another specific embodiment, 15 the antibody can be a plurality of antibodies. The immunologically interacting molecule is specific for Akt kinase or, more specifically, an epitope or epitope on Akt kinase. An epitope or epitope on an Akt kinase includes a portion of the molecule that is guided by an immune response. The antigenic epitope or epitope antigen may be a B cell epitope antigen or, if appropriate, a T cell epitope antigen. In one embodiment, the anti-20 is originally a tyrosine 473 Akt antibody. An embodiment of the present invention provides a method for diagnosing the presence or cancer-like growth of a cancer in a mammal, wherein the Akt activity is abnormal, and the Akt kinase or the antigen thereon is effectively used by Akt kinase binding. An antibody that determines the specificity of a body or a table antigen is contacted with the cell sample obtained from the mammalian cell or the cell extract obtained from the patient, and then quantitatively or qualitatively determined the content of the Akt kinase-antibody complex. The presence of this high content complex compared to normal cells was determined. Any of the 5 10 antibodies known to those skilled in the art can be utilized. For example, in the detection of human Akt kinase, antibodies are usually, but not necessarily, derived from non-human animals, such as primates, livestock animals (eg, sheep, cattle, pigs, goats, horses), laboratory test animals (eg, mice, large Rats, guinea pigs, rabbits) and/or companion animals (eg dogs, cats). Antibodies can also be made recombinantly in prokaryotic or eukaryotic host cells. In general, antibody-based assays can be performed in vitro on cell or tissue biopsies. However, if the right antibody is suitably deimmunized or, for human use, the antibody is humanized, the antibody can be administered, e.g., by nuclear labeling, administered to the patient and the location of the nuclear marker accumulation determined by radiation techniques. The Akt kinase antibody can be a Z-tagged agent. Accordingly, another embodiment of the present invention provides antibodies of such deimmunized forms suitable for imaging cancer in a human or non-human patient. In the case of antibody-producing kinases, if the anti-systems are made by recombinant methods, it is usually necessary to extract the enzyme from the animal (which includes human tissue) or from a cell culture. The Akt can be isolated from the organism by any suitable method. For example, the separation method can utilize any one or more of the following types: surface charge properties, size, density, t activity, and 'Affinity of an entity (eg, another protein shell or chemical compound to which it is associated or associated). Therefore, for example, the fine separation can be separated from the workpiece by any one or more of the following methods: super-exclusion, ion exchange chromatography (for example, anion exchange chromatography, cation exchange chromatography), electrophoresis 66 200831111 (for example, poly Acrylamide gel electrophoresis, isoelectric focusing method), size separation method (such as gel filtration method, ultrafiltration method) and affinity media separation method (such as immunoaffinity separation method, including, but not limited to, magnetic beads Separation methods (such as Dynabead (trademark) separation method, immunochromatography, immunosuppression method). The method for isolating Akt kinase from the '5 biological fluid can maintain the conformational surface antigen present on the kinase and, therefore, It is preferred to avoid the use of techniques that can cause denaturation of the enzyme. In another embodiment, the separation of the peaks from the biological fluid can be performed using any one or more of affinity separation, gel filtration, and/or ultrafiltration. Standard procedures known in the art can be used, for example by Kohler and 10 Milstein (Kohler and Milstein? Nature 256: 495-499, 1975;

Kohler and Milstein, Eur. J. Immunol. 6(7): 511-519, 1976), Coligan et al. ("Current Protocols in Immunology, John Wiley & Sons, Inc., 1991-1997" or Toyama et al. Monoclonal Antibody, Experiment Manual 5, published by Kodansha 15 Scientific, 1987), for the immunological reaction and subsequent preparation of monoclonal antibodies. Basically, animals are cultured with Akt kinases by standard methods. The fluid or a portion thereof or a recombinant Akt kinase is immunized to produce a cell capable of producing an antibody, particularly a somatic cell capable of producing an antibody (e.g., B lymphocytes), which can then be removed from the immunized animal. The cells are immortalized to the cells 20. In a particular embodiment, a fragment of Akt kinase can be used to produce an antibody. The fragment can be associated with a vector. The vector can be native to a non- or low immunogenic material (eg, a hapten) or Any substance that is artificially linked to enhance its immunogenicity, typically of high molecular weight. Can be produced using methods well known in the art to produce 67 200831111 antibody Immortal response of cells. For example, Epstein-Barr virus (EBV) can be used for immortalization of cells (Kozbor et al., Methods in Enzymology 121: 140, 1986). In another embodiment, it is widely used. A cell fusion method for the production of a monoclonal antibody (described in Coligan et al., 199 M997, supra) to immunize the antibody-producing cells. In the present method, an antibody-producing somatic cell having the potential to produce an antibody, particularly The B cells are fused by a myeloma cell line. These somatic cells may be derived from lymph nodes, spleen and peripheral blood of animals that have been exposed to the antigen, preferably to dentate animals such as mice and rats. In this case, mouse spleen cells can be used. In other embodiments, rat, amniotic, sheep or goat cells can also be used. Self-adapted fusion procedures for producing hybridomas (Kohler and Milstein, 1976, supra; Shulman et al, Nature 276: 269-270, 1978; Volk et al, J. Virol 42(1): 220-227, 1982) Lymphoma neoplasms developed a specific myeloma cell line. Many myeloma cells 15 strains can also be used to make fusion cell hybrids, including, for example, P3 multiplied by 63-Ag8, P3 multiplied by 63-AG8.653, P3/NSl-Ag4-l (CN-l), Sp2/0-Agl4, and S194/5.XXO.Bu.l. By Kohler and

Such sp3/〇-Agl4 myeloma 20 cell strain can be formed by multiplying the P3 multiplied by the 63-Ag8 and NS-1 cell strains described by Milstein (1976, supra), Shulman et al. (1978, supra). The sl94/5.XX〇.Bu.1 cell line was reported by Trowbridge (J. Exp. Med. 148(1): 313-323, 1978). A method for producing a hybrid of a spleen or lymph node cell capable of producing an antibody and a myeloma cell usually comprises a ratio of 10:1 in the presence of a drug or a drug group (chemical, viral or electrical) capable of promoting cell membrane fusion, respectively. (Although the ratio can vary from about 2〇·· i 68 200831111 to about i ·· i) mixed somatic cells and myeloma cells. Fusion methods are described (Kohler and Milstein, 1975, supra; Kohler and Milstein, 1976, supm; Gefter et al, Somatic Cell Genet. 3: 231-236 1977; Volk et al., 1982, supra). The fusion agents used by their researchers promoted 5 doses of Sendai virus and polyethylene glycol (PEG). In a particular embodiment, a method of selecting a fused cell hybrid is provided from the remaining unfused cells, particularly unfused myeloma cells. In general, 10 cells of the fused cell hybrid can be cultured in a medium that maintains the growth of the hybridoma but prevents the growth of the unfused myeloma cells, which can generally continue to divide indefinitely. select. Such somatic cells for fusion in in vitro cultures do not maintain long-term viability. It takes several weeks to selectively culture the fused cell hybrids. At the beginning of the period, it is necessary to identify such hybrids which produce the desired antibodies, whereby the hybrids can be subsequently colonized and propagated. In general, about 10% of the obtained hybrids produce 15 antibodies, but ranges from about 1 to about 30% are not uncommon. Hybrids capable of producing antibodies can be detected by any of several standard assay methods, such as, for example, by Kennet et al. (Monoclonal Antibodies and Hybridomas: A New Dimension in Biological Analyses, pp 376-384, Plenum Press , New Yark, 1980) Enzyme-linked 2 plague assay and radioimmunoassay techniques and FACS analysis (O'Reilly et al, Biotechniques 25: 824-830, 1998). Once the desired fusion cell hybridization site has been selected and cloned in individual cell lines capable of producing antibodies, each cell line can be propagated by either of two standard methods. The suspension of such hybridoma cells can be injected into a tissue phase of a tissue phase 69 200831111. The injected animal can then form a tumor that secretes a specific monoclonal antibody produced by the hybrid cell hybrid. Body fluids of the animal, such as serum or ascites fluid, can be released to provide a high concentration of monoclonal antibodies. Alternatively, each of the cell lines can be subjected to in vitro 5 proliferation in a laboratory culture vessel. High concentrations of mono-specific monoclonal antibodies can be collected by decantation, filtration or telecentric methods and subsequently purified. The specificity of the Akt kinase associated with the detection of such cell lines can then be tested by any suitable immunoassay. For example, the cell strain can be divided into several wells and cultured in a plurality of wells, and then the clarified liquid obtained from each well can be analyzed by enzyme-linked immunosorbent assay 10 (ELISA), indirect fluorescent antibody technique or the like. The cell strain (group) producing a monoclonal antibody capable of recognizing the target LIM kinase but not recognizing the non-target epitope antigen can be identified and then directly cultured or injected into a histocompatible animal to form a tumor and be produced and collected. Purify the necessary antibodies. 15 Accordingly, the present invention provides a method for detecting Akt kinase or a fragment, variant or derivative thereof in a sample, which comprises contacting the sample with an antibody or a fragment thereof: a Go derivative and a normal control group (high content thereof) For comparison, to detect the content of the complex containing the antibody and Akt kinase or a fragment, variant or organism thereof, any method capable of determining the formation of the complex can be used, for example, having a related reporter group. The molecules of the antibodies of the invention can be used in immunoassays. Such immunoassays include, but are not limited to, radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), immunochromatography (ICT), and Western blotting methods are well known to those skilled in the art. . The immunization inspection method may also include a competitive verification method. The invention covers both qualitative and quantitative 25 immunoassays. 70 200831111 Suitable immunoassay techniques are described, for example, in U.S. Patent Nos. 4,016,043, 4,424,297, and 4,018,653. These assays include both non-competitive and single-site and dual-site assays as well as traditional competitive assays. These assays also include the direct binding of the labeled antigen-binding molecule 5 to the target antigen. The present invention further provides a method for quantifying the degree of expression or activation of Akt protein in a cell or tissue sample of an animal, such as a human cancer patient or an individual suspected of having cancer. In one embodiment, the invention provides a method for quantitatively quantifying the degree of expression or activation of Akt protein using an imaging system. The imaging system can be used to receive, enhance, and process images of cells or tissue samples stained with Akt protein-specific stains to determine the amount of AKT protein expressed in cells or tissue samples obtained from such animals or The degree of activation. In an embodiment of such methods of the invention, a calibration curve for the expression of AKT1 and AKT2 15 proteins of at least two cell lines exhibiting different levels of AKT protein can be produced. This calibration curve can then be used to quantitatively determine the AKT protein content expressed in a cell or tissue sample. A similar calibration curve suitable for activating the Ακτ protein can be made using reagents specific to these activation properties. It can also be used to determine changes in sputum content and activation status before and after treatment of clinical cancer. In a particular embodiment of one of the methods of the invention, the prion protein expression in a cell or tissue sample can be quantified using an enzyme-linked immunosorbent assay (ELISA) to determine the amount of AKT protein in the sample. Such methods are described, for example, in U.S. Patent Publication No. 2002/0015974. In other embodiments, an enzyme immunoassay can be used to detect the Akt 25 kinase. In such assays, the enzyme complex 71 200831111 is typically combined with a second antibody by glutaraldehyde or periodate. It is generally preferred to use a matrix which can be used in conjunction with such specific enzymes once hydrolyzed by the corresponding enzyme produces a detectable color change. It is also possible to use a matrix which produces fluorescence which produces a fluorescent product rather than a chromogen matrix. The enzyme-labeled antibody can be added to the first antibody-antigen complex to make it adhere, and then the excess reagent is washed away. A solution containing the appropriate substrate can then be added to the antibody-antigen-antibody complex. The matrix can be reacted with an enzyme linked to the second antibody to produce a qualitatively visible signal, which can generally be further quantified by spectrometry to indicate the amount of antigen present in the sample. Alternatively, fluorescent compounds such as luciferin, codamine and lanthanide (铕 10 (EU)) can be chemically coupled to the antibody without altering their binding ability. When activated by illumination of a particular wavelength of light, the fluorescent dye-labeled antibody absorbs light energy, induces an excitable state within the molecule, and then illuminates in a characteristic color that can be visually detected by an optical microscope. The fluorescently labeled antibody is bound to the first antibody-antigen complex. After the unbound reagent 15 is washed away, the remaining tertiary complex is then exposed to light of a suitable wavelength. The observed fluorescence indicates the presence of the relevant antigen. The Immunofluorescence Assay (IFMA) has been well established in this art and is particularly suitable for use in this method. However, other reporter group molecules such as radioisotopes, chemiluminescent or bioluminescent molecules can also be used. 2〇 In certain embodiments, antibodies to Akt kinase can also be used for ELISA vector detection of Akt kinase in serum or other circulating fluids. This is carried out 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 sections. A labeled anti-Akt kinase antibody can then be used to detect Akt kinase as defined in 200872111. This assay can vary in many ways and all variations are within the scope of the present invention and are known to those skilled in the art. The method can be used, for example, in a serum-based assay to rapidly detect and quantify Akt kinase levels. In one embodiment the 'Akt Elisa assay kit can be used in the present invention. Example 5 A Cellular Activation of Signaling ELISA for Akt S473 from SuperArray Bi〇science can be used in the present invention. In one embodiment, the antibody may be an anti-pan antibody that recognizes Akt S473. The Elisa assay kit contains anti-Akt antibodies and additional reagents including, but not limited to, wash buffers, antibody dilution buffers, blocking buffers, cell staining solutions, imaging solutions, stop solutions, secondary antibodies, and Distilled water. Nucleotide Detection In another embodiment, a method for detecting gossin kinase by detecting the degree of expression of a multinuclear > acid encoding an Akt kinase in a cell is provided. Any of the suitable techniques known to those skilled in the art can be used to determine the expression of the 15 polynucleotide. In one embodiment, a labeled polynucleic acid that encodes Akt kinase can be used as a probe in a northern ink spot derived from the cellular iRNA extract. In other embodiments, in a nucleic acid amplification reaction (such as RTPCR), the nucleic acid extract of the animal may be combined with a message relative to the polynucleotide encoding the kinase and an anti-message sequence or a flanking sequence thereof. 20 oxalate primer. Those skilled in the art can use various automated solid phase detection techniques, for example as described by F〇d〇r et al. (Science 251: 767-777, 1991) and Kazal et al. (Nature Medicine 2: 753-759, 1996). Detection technology. In other embodiments, methods are provided for detecting an RNA transcript encoding an Akt kinase. Cellular assays that can be suspected of Akt kinase RNA isolate 25 RNA, for example, from human cancer tissue. RNA can be isolated by the method known in the art, 73, 31,111, for example using TRIZOL reagent (GIBCO-BRL/Life Technologies, Gaithersburg, Md). The first strand of 5 cDNAs can be prepared from the isolated RNA using Oligo-dT or a random sequence nucleotate and sequence-specific oligosole as the primer in the reverse transcriptase reaction. The first cdNAs formed in the PCR reaction can then be amplified by sequence-specific oligonucleotides to produce amplified products. Polymerase chain reaction or "PCR" refers to a procedure or technique in which a large number of pre-selected nucleic acids, RNA and/or DNA fragments are amplified as described, for example, in U.S. Patent No. 4,683,195. In general, sequence information from the ends of the relevant region 10 or outside is used to design oligonucleotide primers. The sequence of these primers is identical or similar to the reverse strand of the template to be amplified. PCR can be used to amplify specific RNA sequences and cDNAs transcribed from total cellular RNA. general

See, for example, Mullis et al. (Quant. Biol. 51:263, 1987; Erlich, eds., PCR Technology, Stockton Press, NY, 1989). Thus, amplification of a specific nucleic acid sequence by PCR 15 relies on oligonucleotides or "introductions" having a reserving nucleotide sequence that is traced back to the arrangement of the relevant gene or protein sequence. For example, sequence comparison of mammalian Akt kinase genes. For example, a primer that is expected to bind to the counter message strand is prepared and another primer that is expected to bind to a 20-mer strand of the cdNA molecule encoding the Akt kinase can be prepared. To detect the amplified product, the reaction mixture is typically subjected to agarose gel electrophoresis or other suitable hybrid technique and the relative presence of the kinase-specific amplified DNA of the Akt is detected. Akt kinase 74 200831111 amplified DNA can be detected, for example, by using a specific oligonucleotide probe for Southern hybridization or by comparing its electrophoretic mobility to DNA standard mobility of known molecular weight. The isolation, purification and characterization of the amplified Akt kinase DNA is carried out by gel excision or elution of fragments (see, for example, Lawn et al, Nucleic Acids Res. 2:6103, 1981; Goeddel et al. Nucleic Acids Res 8:4057-1980), the amplified product is selected in a selection site of a suitable vector, such as the 5 pCRII vector (Invitrogen), sequence analysis of the selected insert is performed and the DNA sequence is compared Known sequences with LIM kinase. The relative amount of LIM kinase mRNA and cDNA can then be determined. In one embodiment, time-consuming PCR can be used to determine the Akt nuclear oxalate conversion procedure. The determination of transcriptional activity also includes the determination of potential transcriptional activity based on the availability of mRNA transcripts. Real-time PCR and other PCR programs use many chemical methods that can be used to detect PCR products, including DNA-binding fluorophores (5' endonuclease), adjacent lining and hairpin oligonucleotide probes, and spontaneous fluorescence A combination of amplicons. These chemical methods and real-time PCR are discussed in the following literature: Mackay et al., Nucleic Acids Res 30(6): 1292-1305, 2002; Walker, J. Biochem. Mol.

Toxicology 15(3): 121-127, 2001; Lewis et al., j. path〇i· 195: 66-71, 200. In another embodiment, the abnormal expression of Akt is identified by having 20 A nucleus citrate probe capable of complementing a sequence of an Akt sequence selected from a nucleotide sequence of the 6a-c, 7a-d or 8a-c or a fragment thereof, is contacted with a nuclear sulphate isolated from a biological sample The sequence is then detected by hybridizing the probe for the sequence and comparing the results to the normal sample. Hybridization of the probe for the biological sample can be detected by labeling the probe with any detectable agent. 75 200831111 The probe may be labeled with, for example, a radioisotope, a biotin, a fluorescent dye, an electronic cryptic agent, an enzyme, a hapten or a protein that can be utilized. The detectable 5 degree can be evaluated in any desired manner, including spectrometry, photochemical, biochemical, immunochemical, radioisotope or chemical methods. The probe can also be used to detect such probes: such as oligomer restriction techniques, dot ink assays, reverse dot ink assays, multi-column probe assays, and 5, nuclease assays. Alternatively, any of a wide variety of widely used DNA array technologies can be used, including giant arrays, microarrays, and DNA microchip technology to detect the probe. The oligonucleotide probe typically comprises about at least 14, 15, 16, 18, which can hybridize to a nucleotide selected from the group consisting of 6a-c, 10 7a-d, and 8a-c or a fragment thereof. 20, 25 or 28 nuclear oxalate. It is generally preferred not to use probes that are greater than about 25 or 28 nucleotides in length. This oligonucleotide probe is used to determine the Akt nucleotide sequence. Kinase assays 15 Any suitable kinase assay known in the art can be used to determine the activity of such Akt kinases. For example, Hogg et al. (Oncogene 1994 9: 98-96), Mills et al. (J. Biol. Chem. 1992 267: 16000-006) and Tomizawa et al. 2001 (FEBS Lett. 2001 492: 221-7) can be used. The method described in Schmandt et al. (J. Immunol. 1994, 152: 96-105), but is not limited thereto. In addition, the serine, threonine and tyrosine kinase assays are described in Ausubel et al. (Short Protocols in Molecular Biology, 1999, unit 17-6).

Akt kinase assays typically employ an Akt polypeptide, a labeled donor matrix, and a receptor matrix that is specific or non-specific for Akt. The kinase is determined by the number of labeled molecular groups in which the Akt can transfer a labeled molecular group from the donor matrix to the acceptor base and transfer from the donor matrix to the acceptor matrix in the method of 2008 2008111111. active. A variety of expression systems can be used to make Akt polypeptides which can be purified from cells, can be in the form of a split or unsplit recombinant 5 fusion protein and/or can have a non-Akt polypeptide sequence, for example, in its N- or C - The end has a His tag or a galactosidase. If a cancerous cell line is used as a source of Akt to be assayed, Akt activity can be assayed in these cancerous cell lines. A donor matrix suitable for Akt assays includes any molecule that is susceptible to dephosphorylation by Akt, such as labeled Ατρ and Ατρ analogs, 10 wherein the label is 33P, 32P, 35S or any other radioisotope or suitable fluorescing mark. Acceptor matrices suitable for Akt assays include any polypeptide or other molecule that is susceptible to phosphorylation by Akt. The receptor matrix can be derived from a fragment of an in vivo target of Akt. The acceptor matrix fragment may be 8 to 5 amino acids in length, typically 1 to 3 amino acids, and preferably has about 1 , 12, 15, 1 5 18, 20 and 25 amino acids. . A range of different peptides or other molecules can be used empirically to determine additional receptor matrices. Once the reaction is carried out, the target of the acceptor matrix suitable for TTK can be purified from the other components of the reaction. The purification reaction is usually carried out by molecular interaction, wherein the receptor matrix is subjected to biotinylation reaction and by using it with streptavidin 2〇 or a specific antibody which can be used to uniquely identify the receptor substrates Purification by interaction. The reaction can be carried out under various conditions, such as on a solid carrier, in a gel, in a solution, or in a living cell. The choice of detection method depends on the type of label used for the donor molecule and may include, for example, hybrid autoradiography, flash, scanning, or fluorescence radiography. . Methods of Treatment The compounds and pharmaceutical compositions provided herein can be used to treat a condition, including tumors, cancer, and other conditions associated with abnormal cell proliferation. 5 In one embodiment, the compounds of the invention are useful for treating cancer, sarcoma, lymph, white disease, and/or myeloma. In other embodiments of the invention, the compounds disclosed herein can be used to treat solid tumors. The compounds of the invention are useful in the treatment of cancer, such as, but not limited to, cancer of the following organs or tissues: breast, prostate, lung, bronchi, colon, urinary system, bladder, non-Hodgkin's lymphoma, melanoma , renal tumor, kidney system, pancreas, pharynx, thyroid, stomach, brain, multiple myeloma, esophagus, liver, intrahepatic biliary tract, neck, larynx, acute myeloid leukemia, chronic lymphatic leukemia, soft tissue (such as heart) , Hodgkin's lymphoma, testicular, small intestine, chronic spastic leukemia, acute lymphocytic leukemia, anal, anal canal, anal 15 and rectal system, thyroid, female genital, gallbladder, pleura, eye, nose, nasal cavity, Middle ear, nasopharyngeal, ureter, peritoneum, omentum, mesentery, and gastrointestinal tract, high degree glioma, glioblastoma, colon, rectum, pancreas, stomach cancer, hepatocellular carcinoma; head and neck cancer, carcinoma Renal cell carcinoma; adenocarcinoma; sarcoma; hemangioendothelioma; lymphoma; leukemia, verrucous granuloma. In other embodiments, the compounds of the invention may be used to treat dermatological conditions including, but not limited to, the following malignant diseases: angiosarcoma, hemangioendothelioma, basal cell carcinoma, squamous cell carcinoma, malignant melanoma, and card Persist's sarcoma; and non-malignant diseases or conditions, such as psoriasis, lymphangiogenesis, hemangioma in children, Sturge-Weber syndrome, 78 200831111 gynecological night fiber, scorpion facial tumor, nodular sclerosis , purulent granuloma, recessive dystrophic large foamy epidermis loose, venous ulcer, hemorrhoids, rosacea, ', treatment pass 1 ± soft night, sebum leakage keratosis, and actinic keratosis. Compositions comprising such compounds of the invention may be used to treat these cancers and other cancers from any stage of the discovery of the cancer 5 to the advanced stage. In addition, compositions comprising the compounds of the invention may be used to treat primary cancer and its metastasis. In other embodiments of the invention, the compounds described herein are useful for treating cancer, including, but not limited to, the cancers disclosed in Table 1 below. □ Table 1: Cancer type □ Adult acute platinum disease Child acute lymphoblastic leukemia Adult acute myeloid leukemia Child Acute mineral leukemia Adrenal cortical cancer Child Adrenal cortical cancer AIDS Related cancer AIDS Associated lymphoma Anal cancer Child Cerebellar star Astrocytoma children cerebral astrocytoma □ basal cell carcinoma extrahepatic biliary tract cancer bladder cancer children bladder cancer bone cancer osteosarcoma / malignant fibrous histiocytoma children brain stem glioma adult brain tumor children brain tumor, Brain stem

□ Hair cell leukemia head and neck cancer Adult liver cell (liver) cancer (primary) Child liver cell (liver) cancer (primary) Adult Hodgkin's lymphoma child Hodgkin's lymphoma during pregnancy Jijin's lymphoma hypophal carcinoma children's hypothalamus and optic nerve bundle glioma □ intraocular melanoma islet cell carcinoma (endocrine pancreas) □ Kaposi's sarcoma (Kaposi, s Sarcoma) kidney (kidney cell) cancer children's kidney Cancer, laryngeal cancer, laryngeal lymphoblastic cell leukemia, f-lymphoma, myeloid leukemia, myeloid leukemia, 79, 3131, 2011, glioma, cerebellar brain tumor, astrocytoma, cerebral cerebral tumor, astrocytoma, malignant nerve Glioma children brain tumors, ependymoma children brain tumors, neural tube blastoma on-screen brain tumors children primitive neuroectodermal tumors children brain tumors, optic nerve bundles and hypothalamic glioma children brain tumor breast cancer Childhood breast cancer, male breast cancer, bronchial adenoma, carcinoid Burkitfs Lymphoma, pediatric carcinoid gastrointestinal carcinoid tumor Known primary central nervous system cancer children cerebellar astrocytoma children cerebral astrocytoma / malignant glioma children glioma cervical cancer children cancer chronic lymphocytic leukemia chronic spinal hyperplasia disease colorectal cancer children Colorectal cancer cutaneous T-cell lymphoma, see verrucous granuloma and zeazel syndrome chronic lymphocytic leukemia chronic inflammatory leukemia B cell leukemia lip and oral cancer adult liver cancer (primary) child liver cancer (primary) non-small Cell lung cancer small cell lung cancer AIDS-associated lymphoma Burkitt's lymphoma cutaneous T-cell lymphoma, see verrucous granuloma and Szezly syndrome (S0zary) syndrome adult Hodgkin's lymphoma children Hodgkin's lymphoma pregnancy Hodgkin's lymphoma adult non-Hodgkin's lymphoma children non-Hodgkin's lymphoma pregnancy non-Hodgkin's lymphoma primary central nervous system lymphoma □ macroglobulinemia, bone Waldenstr0m's malignant fibrous histiocytoma/osteosarcoma children's neural tube blastoma melanoma Tumor (eye) melanoma, Merkel cell carcinoma, adult malignant mesothelioma, mesothelioma, recessive primary metastatic squamous cell carcinoma, multiple endocrine neoplasia syndrome, multiple myeloma/plasma Cellular squamous granuloma myeloproliferative syndrome myelodysplastic/myeloproliferative 80 200831111

Children's ependymoma esophageal cancer children esophageal cancer Ewing's family tumor children gonad blasts tumor gonad blasts tumors extrahepatic squamous cell carcinoma of the eye, intraocular melanoma eye cancer, retinoblastoma □ Gallbladder cancer Gastric cancer Childhood Gastric cancer Gastrointestinal carcinoma Gu blast cell tumor Childhood Glandular blast cell tumor Ovarian blast cell tumor Gestational trophoblastic tumor Adult glioma Child brain stem Glioma Child cerebral glioma Astrocytoma Children Vision nerve bundle and hypothalamic glioma □ skin cancer (melanoma) Mock cell skin cancer small cell lung cancer small bowel cancer adult soft tissue sarcoma children soft tissue sarcoma squamous cell carcinoma, see skin cancer (non-melanoma) stability Primary metastatic squamous cell carcinoma, gastric cancer, children with gastric cancer, supratentorial primitive chronic myeloid leukemia, acute myeloid leukemia, childhood myeloid leukemia, malignant myeloma, malignant myelodysplastic disease, paranasal sinus cancer, nasopharyngeal Cancer children, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, child Non-Hodgkin's lymphoma, non-Hodgkin's lymphoma, non-small cell lung cancer during pregnancy, children's oral cancer, I cavity cancer, lip and oropharyngeal cancer, osteosarcoma, bone malignant fibrous histiocytoma, ovarian cancer Nest epithelial cancer egg blast cell tumor ovarian low malignant potential tumor □ pancreatic cancer child pancreatic cancer, island cell pancreatic cancer sinus and nasal cancer parathyroid carcinoma yin cancer pheochromocytoma Cell tumor and supratentorial primitive neuroectodermal tumor pituitary tumor plasma cell tumor / multiple myeloma pleural and lung blastoma pregnancy breast cancer pregnant period Hodgkin's lymphoma pregnancy non-Hodgkin's lymph, System 81 200831111

Verrucous granuloma and Xizelai syndrome group sputum cancer children thymoma thymoma and thymic carcinoma thyroid cancer children thyroid cancer sputum and ureteral migration cell carcinoma surname yin trophoblast tumor □ adult unknown primary European cancer Unknown primary site of cancer in children, children with ureter and renal pelvis, abnormal cancer, transitional cell carcinoma, urethral cancer, endometrial uterine cancer, uterine sarcoma, vaginal cancer, children's optic nerve bundle, and hypothalamic glioma, female squamous cell carcinoma Wilm's Tumor, prostate cancer, renal cell (kidney) cancer, renal cell (kidney) cancer, stenosis and ureteral transitional cell carcinoma, retinoblastoma, striated muscle Sarcoma salivary gland cancer, salivary gland carcinosarcoma, eus family tumor, Kaposi's sarcoma, adult soft tissue sarcoma, uterine sarcoma, zeezel syndrome, skin cancer (non-melanoma), child skin cancer, in another embodiment of the invention, The compounds disclosed herein can be used to treat angiogenesis-associated diseases. Angiogenic small molecules include thalidomide, which has five parts that inhibit NFkB; 2-methoxyestradiol, which affects microtubule activation and hypoxia-inducible factor (HIF la) Activation; cyclooxygenase 2 (COX2) inhibitors; and low doses of conventional chemotherapeutic agents, including cyclophosphamide, anthracycline analogs, and vinca alkaloids (vincristine) Vinblastine (D, Amato, RJ et al. 82 200831111 (i994) Proc. Natl. Acad. Sci. USA91, 3964-3968, D, Amato? RJ· et al. (1994) Proc· NatL Acad · Sci. USA91, 4082-4085). In addition, specific tyrosine kinase inhibitors can indirectly reduce angiogenesis by reducing VEGF and other pro-angiogenic factors produced by tumors and stromal cells. Hexceptin, imatinib, (Ghvec), and Iressa (Bergers, G et al. (2003) are 0/ /πνα% as (10) ill, 1287-1295, Ciardiello, F. et al. (2001) Clinical Cancer Research 7, 1459-1465, Plum, SM Specialist (2003) 9, 4619-4626) o 10 Recent ' Tube-forming inhibitors have evolved into animal models. The blood-forming inhibitors represent promising treatments for various cancers. Recently, arvastatin, which is an anti-vascular endothelial growth factor (VEGF), It has been shown to be a single agent in advanced renal cell carcinomas to prolong life in advanced colon cancer and to use chemotherapy to prolong life (Yang, JC et al. 15 (2003) New England Journal of Medicine 349? 427-434,

Kabbinavat, F. et al. (2003) journal 〇f Clinical 〇nc〇1〇gy 21, 60-65) Related diseases of the sputum include, but are not limited to, inflammation, autoimmune disease and sensation of wood disease, Angiogenesis-dependent cancers include, for example, solid tumors, 2 consecutive tumors (such as leukemia), and tumor metastasis; good tumors such as blood & tumor 1, see God, neurofibromatosis, trachoma, and purulent granulation Swollen; rheumatoid arthritis; psoriasis; moist treatment; ocular angiogenic diseases, such as diabetic retinopathy, premature retinopathy, macular degeneration, t 4 rejection of neovascular glaucoma, increased fibrous tissue after lensing 83 200831111 Redness; Osler-Weber syndrome; myocardial angiogenesis, plaque new blood production; microvascular dilatation; hemophilic joints; angiofibroma; and wound granulation. In addition, the compositions of the present invention may be used to treat diseases such as, but not limited to, intestinal adhesions, atheroma hardening, night, and 5 hypertrophic scars (i.e., tumors). The compositions of the invention may also be used to treat diseases which have angiogenesis due to pathogenic consequences, such as R0chele minalia qmntosa, Helobacter pyl〇ri, tuberculosis, and leprosy. Therapeutic Tumors or Pain Relief + The present invention provides compounds useful for the treatment of drug-resistant cancers, including the cancer and triclinibine compounds disclosed in Figure 10 and/or such onioncycline analogs. Multidrug resistance (MDR) occurs in human cancer and can be a major obstacle to the success of chemotherapy. Multidrug resistance is a phenomenon in which tumor cells exposed to cytotoxic agents form cross-resistance to a range of structurally and functionally unrelated compounds. In addition, MDR can be endogenous in some cancers that were not exposed to chemotherapeutic agents before 15 . Accordingly, in one embodiment, the invention provides a method for 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 Or a variety of onioncycline analogs. In a particular embodiment, TCN, TCN-P, and related compounds and one or more onionine analogs can be used to treat 2 〇 only for Taxol, rapamycin, Tamoxifen, Cisplatin, and/or gefitinib (Aressa) resistant cancer. In one embodiment, TCN, TCN-P or related compounds and one or more onioncycline analogs as disclosed herein can be used to treat the following drug-resistant cancers: colon, bone, kidney, adrenal gland, pancreas, liver, and / or any other cancer described in this technique 84 200831111. Known in the art or in combination with a towel, in one embodiment, the compound can be called I 'benfax _# 曲 赛 滨 化合物 化合物 化合物 化合物 化合物 化合物 化合物 化合物 化合物 化合物 化合物 化合物 化合物 化合物 化合物 化合物 化合物 化合物. In another embodiment, etc., the composition of the compound and trastuzumab as astuzumab) and in the case of the stomach for use in the treatment of solid tumors, the use of radiation

In another embodiment of the invention, the escitalin and the auxin compound and composition disclosed herein may be used in combination with at least one additional chemotherapeutic agent. The additional agents may be administered together with the compounds disclosed herein or by transfusion - the drugs may form part of the same composition or may be provided in individual compositions suitable for administration at the same time or at different times. In one embodiment, the trehalin compound and the lycopene a substance disclosed herein may be combined with an anti-angiogenic agent to enhance their potency, or 15 may be combined with other anti-angiogenic agents and sterilized with other cells. Sex agents are administered together. In other embodiments, when used to treat a solid tumor, the tresibine compound and the onionine compound may be administered together with an agent selected from the group consisting of IL-12, retinoic acid. (retinoids), interferon, vasostat, angiostatin, endostatin, diazepam, thrombospondin-1, thrombospondin-2, captopril (captopryl), anti-tumor agents (such as alpha interferon), COMP (cyclophosphamide, periwinkle, methrexate and prednisone), etoposide ), mBACOD (american, bleomycin, cranberry, 85 200831111 ring-filled guanamine, periwinkle new test and dexamethasone (dexamethasone), PRO-MACE/MOPP (prednisone) , Ami Sujing (w/ kecofu (leucovin) rescue agent), cranberry, cyclophosphamide, scorpion scorpion / mechlorethamine, vinca alkaloid, procarbazine), Changchun Huaxin base, 5 vinblastine, angiostatin (antioinhibins), TNP-470, polypentose (pentosan) , polysulfate, platelet factor 4, vasstatin, LM-609, SU_101, CM-101, Techgalan, Shali Doumai, SP-PG, and radiation. In other embodiments, the disclosure The compound and composition may be administered together or in a rotating manner: for example, a drug having anti-mitosis for 10, such as a drug capable of calibrating an element of a cytoskeleton, including podophylotoxin or vinca alkaloid (vincristine, vinca); antimetabolite drugs (such as 5-fluorouracil, cytarabine, gemcitabine, purine analogs, such as pentostatin, Amesu烷); alkylating agent or nitrogen mustard (such as leucouret, cyclamate or ifosphamide); drugs that can be labeled with DNA, such as the anthracycline drugs, leucomycin (adriamycin), cranberry, pharmorubicin or epirubicin; a drug that can be labeled with topoisomerase, such as etoposide; hormones and hormone agonists or antagonists, Such as estrogen, anti-estrogen (temoxifen and Compounds and males 2 hormones, flutamide, leuprorelin, goserelin, cyprotrone or octreotide; signal transduction in tumor cells Labeled drugs, including antibody derivatives, such as oxetine; alkylating drugs, such as platinum drugs (cisplatin, carboplatin, oxaliplatin, paraplatin or 86 200831111) Nitrourea; a drug that can potentially affect tumor metastasis, such as matrix metalloproteinase inhibitors; gene therapy agents and anti-message agents; antibody therapeutic agents; other biologically active compounds of marine origin, especially didemnin, such as Apo Aplidine; a steroid analog, especially dexamethasone; 5 an antiemetic, including a 5HT-3 inhibitor (such as gramisetron or ondasetron, with steroids and Derivatives such as dexamethasone. In still other embodiments, the compounds and compositions can be used with or in turn with the chemotherapeutic agents disclosed in Table 2 below. Table 2: chemotherapeutic agent --------13-cis-retinoic acid-2-amino-6-Weiylindole-2-CdA-2-gas deoxygenated gland-5-gas urinal π定-5-FU -6 - TG -6-sulfostigmine-6-weiki嘌呤-6_MP -Accutane - actinomycin-D - Ai Huisu - Adrucil - Ann Agrylin - Ala-Cort - Aldesleukin - Alemtuzumab - Alitretinoin - Alkaban - AQ - Wicker Alkeran) - All-trans retinoic acid-α-interferon-Altretamine-Neosar-Neulasta-Neumega-Neupogen- Nilandron - Nilutamide - Novaldex - Novantrone - Octreotide - Octreotide Acetate - Oncospar - Ontak - Oncology Onxal - Oprevelkin • Orapred - Orasone - Oxaliplatin - Pacific Paclitaxel - Pamidronate - Panretin -Pediapred 87 2o〇8311ii - Amethopterin - Amine Phosphorus Amifostine - Aminoglutethimide 'Anagrelide' - Anandron "Anastrozole -Acytosylcytosine -Allah Ara)-C -Aranesp -Aredia -Arimidex -Aromasin - Arsenic Trioxide Aspartate - ATRA - Avastin -BCG -BCNU -Bevacizumad -Bexarotene -Bicalutamide -BiCNU Blenoxane -Bleomycin -Bortomycin (Bortezomib) - Busulfan - Busulfex - C225 - Calcium Leucovorin - Campos - Camptosar - Camptothecin )-11 - Capecitabine - Carac - Carboplatin - Carmustine - Carmustine Wafer - Casodex - CCNU - PEG Interferon - Pegaspargase - Pegfilgrastin PEG Insertion (Intron) -PEG-L-Aspartame-Phenylalanine-Phosphate-Platinol-Phosphoric Chloride amine -AQ -Prednisolone -Prepone -Prokrit - Proleukin -Pfizers with Carmustine Implants (Prolifeprospan) 20 - Purinethol - Raloxifene - Rheumatrex Rinuxan - Rituximab - Roveron -A (a-2a interferon) - Rub ex - Rubidomy cin hydrochloride - Sandostatin - Good LAR - Sargramostim Solu-Cortef - Solu-Medrol - STI-571 - Streptozocin - Tamoxifen - Targretin (Taxol) - Taxotere - Temodar - Temozolomide - Teniposide 88 200831111

-CDDP -CeeNU -Cerubidine _Cetuximab -Chlorambucil •Cisplatin -Citrovorum factor -Cladribine - Cortisone - Cosmegen -CPT-11 -Cytadren - Cytadren Cytarabine - Cytarabine -Cytosar )-U - Cytoxan - Daccarbazine - Dactinomycin - Darbepoetin alfa • Daunomy cin 'Daimombicin 'Danobicin hydrochloride-Danobiocin liposome-DaimoXome-Decadron-5-Delta-Cortef-Deltasone-Dinisin Denileukin difitox) - DepoCyt - Dexamethasone "• Dexamethasone Acetate Samidt Sodium - Dexasone - Dexrazoxane -DHAD -DIC -TESPA Thalidomide - Thalomid - TheraCys - Thioguanine - Thioguanine Pill - Thiophosphamide - Sepp Thioplex - Thiotepa - TICE - Toposar • Topotecan - Toremifene - Trastuzumab - Vitamin Octaic Acid (1^^ 11〇1|1) -Trexall -Trisenox -TSAP -VCR -Velban -Velcade -VePesid -Vesanoid ) -Viadur - Changchun Flower Test - Vinca Base Sulfate Salt - Vincasar Pfs Long Fragrance New Test - Venofolbine - Venokine Tartrate - VLB - VP-16 - Vumon - Xeloda - Zanosar - Zevalin - Zinecard - Zoladex - Zoledronic acid 89 200831111 -Diodex -Docetaxel -Doxil -Doxorubicin -Cranberry Dlipid -Droxia -DTIC -DI1C-Hemisphere - Duralone - Efudex - Eligard - Ellence - Eloxatin - Elspar - Emcyt - Table Epinubicin -α Epoetin alfa - Erbitux • Io Erwinia L-aspartate - Estramustine - Ethyol - Etopophos - Sneaky Alfalfa - Etoposide Phosphate Eulexin - Evista - Exemestane - Fareston - Faslodex - Femara - Filgrastin - Say Fluxuridine - Fludara - Fludarabine - Fluoroplex - Fluorouracil - Fluorescent Cancer Star - Emulsion - Fluoxymesterone - Zometa - Gliadel wafer • Glivec - CM-CSF - Goserelin - Granulocyte - Colony Stimulating Factor Cell Macrophage Colony Stimulating Factor - Fluoride Halotestin - Hepatotine - Liu Zhuolong, Hexadrol _ Hexalen - Hexamel melamine - HMM - Hycamtin • Hydroea - Hydrocort Acetate • Hydrocorticone (hydrocorcortisone) - Hydrocorticosterone Sodium Phosphate &Hydrochloride; Hydrocorticosterone Succinate - Hydrocorticosterone Phosphate • Light-based Urea • Ibritumomab - Tiemo Monoclonal antibody (Ibritumomab Tiuxetan) - Idamycin, Idarubicin • Ifex • a -IFN - Ifosfamide - IL^2-IL- Li -Imatinib mesylate-imidazole carboxamide interferon•a-2b interferon (PEG conjugate) - baibaisu-2 - baibaisu-11 beta seeding (Introm ) A(a -2b interferon) - ercofu 90 200831111

- Flutamide

-Folinic Acid -FUDR -Fulvestrant -G-CSF -Gefitinib -Jessitabine - Gemtuzumab ozogamicin -Gemzar - Gili Gleevec - Lupron - Lupron Depot - Matlane - Maxidex - Mechlorethamine - Mecamylamine - Medralone -Medrol -Megace - Megestrol - Megestrol acetate - Melphalan - Seki σ 呤 - Mesna - Beauty Mesnex _ Amesu sulphate - Amesu sulphate - methyl adrenocortic ketone - Mylocel - Leukeran _ Leukine - Liu Peilin " Changchun New test (Leurocristine) "Lusit stop (Leustatin)

• Liposomal Ara-C - Liquid Pred - Lomustine "L-DAM -L-Sharc〇iySin - Meticorten -Mitomycin-Mitomycin-C-Mito Onion

Prednisol - MTC -MIX • Nistar mustard (Mustargen) - Mustard, Mustine - Mutamycin - Myleran - Aretha Elaine Dickon Irinotecan) - Isotretinoin - Kidrolase

- Lanaket-L-aspartate -LCR - Letrozole - In a particular embodiment, interferon (IFN) can be used with the compounds of the invention. Suitable interferons include: _2a interferon, a-2b interferon, pegylated alpha interferon (which includes _2a interferon and alpha-2b interferon), /3 interferon, 7 interferon, τ interference , ω interferon, INFERGEN (alphacon-l interferon) manufactured by InterMune, OMNIFERON (natural interferon) manufactured by Virage 91, 200831111, and Russian Buffer made by Human Genome Science ALBUFERON, REBIF (cold-1 a interferon) manufactured by Ares-Serono, omega interferon manufactured by BioMedicine, oral interferon alpha manufactured by Amarillo Biosciences, and interferon manufactured by InterMime 5, τ interferon, and / or r-1/3 interferon. In one embodiment, a TCN, TCN-P or related compound and an onionine compound as disclosed herein may be used in combination with another chemotherapeutic agent, such as the chemotherapeutic agents described herein or in Table 2, to treat drug-resistant cancer. For example, multidrug resistant cancer. Drug resistant cancers can include colon cancer, bone cancer, kidney cancer, 10 adrenal cancer, pancreatic cancer, liver cancer, and/or any other cancer known in the art or described herein. In one embodiment, the additional chemotherapeutic agent can be a P-glycoprotein inhibitor. In a specific non-limiting embodiment, the p-glycoprotein inhibitor may be selected from the group consisting of verapamil, cyclosporine (such as cyclosporine A), temoxim, calcium Protein kinase (calm〇dulin) antagonizes 15 doses, dexverapamil, dexniguldipine, vaiSp〇dar (PSC 833), biricodar ( VX-710), tariquidar (XR9576), zosuquidar (LY335979), laniquidar (Rl〇i933), and/or ONT_093. 20 Pharmaceutical Compositions These compositions comprising a tripamicin compound and one or more anthracycline analogs can be optionally administered with a pharmaceutical carrier or excipient. Pharmaceutical carriers suitable for administration of the compounds provided herein include any such carriers known to those skilled in the art to be suitable for the particular mode of administration. The esculin compound 92 200831111 and one or more cycline analogs may be formulated together into the only active ingredient in the composition or may be combined with one or more anthracycline analogs. Compositions comprising the tresibine compound and one or more onioncycline analogs may be suitable for oral, rectal, nasal, topical administration (including 5-frequency and sublingual administration), vaginal administration or parenteral administration (It includes subcutaneous, intramuscular, intravenous, intradermal, intraocular, intratracheal, intracisternal, intraperitoneal, and epidural) administration. Preferably, the compositions are administered intravenously. These compositions may preferably be in unit dosage form and may be prepared by conventional pharmaceutical techniques. Such techniques include associating one or more compositions of the invention with one or more 10 pharmaceutical carriers or excipients. The citrifloxacin compound and one or more onioncyclin analogs and compositions thereof may be formulated into a pharmaceutical preparation suitable for oral administration, such as a solution, a suspension, a lozenge, a dispersible lozenge, a pill, A capsule, a powder, a sustained release formulation or an elixir, or a sterile solution or suspension 15 suitable for parenteral administration, and a transdermal patch and a dry powder inhaler. In one embodiment, the above-described triclinic compound is converted into a musical composition using techniques and procedures well known in the art (see, for example, Ansel Introduction to

Pharmaceutical Dosage Forms, 4th edition, 1985, 126). In such compositions, one or more of the effective concentrations of Compound 20, or a pharmaceutically acceptable derivative thereof, can be combined with one or more suitable pharmaceutical carriers. The compound of the present invention can be derivatized into a corresponding salt, ester, dilute alcohol ether or ester, acetal, ketal, orthoester, hemiacetal, hemiacetal, acid, alkali, / granule, hydration before preparation. Or prodrug. Once administered, the concentration of such compounds in the composition is effective to deliver an amount that can treat, prevent, or ameliorate the target disease or the symptoms of the disease of 93 200831111. In the actual - 5 15 20, the money is modulated in order to adjust the limbs, so that: the dissolved compound dissolves, _, disperses the sister in the specific carrier H concentration can effectively transfer, prevent the treatment of the disease or can improve a symptom. The group suitable for oral administration can be used for the description of the money: individual but not limited to the amount of each ingredient - ❹ (4) community = agent, caplet, pill or dragee, capsule or capsule; powder or Granules; assays or stock solutions in aqueous or non-aqueous liquid wipes; or oil-in-water liquid emulsions or water-in-oil emulsions, or doughs. For example, by dissolving, dispersing or mixing tromethine and optionally selecting a pharmaceutical adjuvant to form a solution by using a carrier such as water, a saline solution, aqueous dextrose, glycerin, ethylene glycol, ethanol, or the like (IV) The suspension is made into a liquid pharmaceutically acceptable composition. If necessary, the pharmaceutical composition to be administered may also contain small amounts of non-toxic auxiliary substances such as wetting agents, emulsifiers, solubilizers, pH buffers, preservatives, flavoring agents, and the like, such as acetate, citric acid Sodium, cyclodextrin derivatives, sorbitan monolaurate, triethanolamine acetate, triethanolamine oleate, and other such agents. Methods of preparing such dosage forms are known or known to those skilled in the art; for example, see

Remington s Pharmaceutical Sciences, Mack Publishing

Company, Easton, Pa., 15th edition, 1975. Compositions of the invention suitable for topical administration in the oral cavity include, for example, flavours-based ingredients, typically sucrose and acacia gum or tragacanth; one or more of the triclinide compounds of the invention and one Or a variety of onioncycline analogs in inert ingredients such as gelatin and glycerin, or sucrose and sulphate 94 200831111; and sputum having ~ or more of the compositions of the invention administered in a suitable liquid carrier Mouth liquid. Such lozenges, pills, capsules, buccal tablets and the like may contain ~ or more compounds having a composition of 5 10 15 20 or the like: a binder, a lubricant, a diluent, a slip aid, a decomposing agent, a coloring agent. , sweeteners, flavoring agents, slip agents, emetic coatings, and film coatings. Examples of the binder include microcrystalline cellulose, carrageenan, glucose solution, gum arabic mucilage, gelatin solution, glycocalyx, κ 乙 唆 唆 唆, polyethylene 吼 目 同 ( ( ( ( ( ( ( ( ( ( (联之|乙浠口比咯_, sucrose and starch paste. Lubricants include talc, ,,, sputum, stearin # or hard yue 35,; 6 pine nuts (lyeGpGdiu 啦 hard. Thinners include, for example, lactose, Yan sugar, starch, kaolin, salt, mannitol and: acid = °The slip agent includes 'but is not limited to: Oxygen dioxide. The decomposing agent includes the sodium carboxymethyl cellulose, sodium starch glycolate, Haidosa, the temple powder, the horse mash, the soil, and the Kelin, material and cellulose. Colorants include, for example, any approved water-capacity and C dyes, mixtures thereof; and water suspended on oxidized hydrates:::: FD and C dyes. Sweeteners include Yan sugar, lactose, mannitol and = sweeteners, such as saccharin, and any amount of dry-reading flavoring agents. ^ Agents include days extracted from plants (_fruits) A synthetic compound blend that produces 2 sensations, such as, but not limited to, peppermint and willow. The genus includes propylene glycol monostearate, sorbitol needle monooleate, diethylene glycol single month _ and poly Oxidized acetonitrile _. vomiting coating = fatty acid, m _, worm (tetra) acid cellulose 酞i 曰. binding film coating bag Hydroxyethyl cellulose, sodium carboxymethyl cellulose, poly 95 200831111 ethylene glycol 4000 and cellulose acetate phthalate. _ suitable for local administration of money. (4) with - or multiple can be connected on the table A soft f, emulsion, _, and paste preparation of the composition to which the reader is administered. The composition may be provided as a suppository having a suitable base L % 匕-I λ » », for example, cocoa butter or salicylate. When the carrier is a solid, the composition suitable for nasal administration comprises a coarse agent which can be administered by inhalation (i.e., by inhalation through the nasal passages from the container close to the nose). The particle size may be, for example, in the range of micrometers - or a plurality of such compositions may be incorporated into or absorbed into the nasal passages in an aqueous or oily solution. Compositions suitable for vaginal administration may contain one or more of such materials. And a suitable carrier vaginal suppository, tampon, emulsion, gel, paste, foam or spray. 15 compositions for parenteral administration include antioxidants, buffers, bacteriostats, And making the formulation isotonic with the blood of the predetermined recipient Aqueous and non-aqueous sterile injectable solutions of solute; and aqueous and non-aqueous sterile suspensions which may include suspending agents and granules. These compositions may be in unit dose or multi-dose containers (eg sealed ampoules and vials) Provided and can be stored in only 2 〇 required to add liquid carrier shortly before use, such as cold water for injection; East dry (freeze-dried) conditions can be made from the previously described sterile powders, granules and key types Ready-to-use injections and suspensions. The compositions can be prepared using a pharmaceutically acceptable organic or inorganic solid or liquid carrier medium suitable for enteral or parenteral administration. Gelatin, lactose, starch, 96 200831111 Recording, diol, water or: = vegetable and animal fats and oils, gums, poly-extensions, and known carriers are all suitable as carrier agents. Such as: medium: a variety of pharmaceutically acceptable carrier media and / or shaping is suitable for & </ "> pharmaceutically acceptable carrier media f, including any and

10 agents for specific dosage forms of carriers, solvents, diluents or other liquid vehicles, 4 suspension adjuvants, surfactants, isotonic agents, pastes or milk systems = rot, solid binders, lubricants, Adjuvant, vehicle, delivery sweetener: decomposed absorbent, surfactant, colorant, flavor or

In addition, the compositions comprising the triclinide compound and the one or more onioncyclines may be combined with a pharmaceutically acceptable excipient and, if desired, a release matrix, such as a biodegradable polymer, to form Therapeutic Composition 15 pharmaceutically acceptable excipients, including non-toxic solid, semi-solid or liquid fillers, diluents, encapsulating materials or formulation adjuvants of any type. However, it should be understood that the total daily usage of the constituents is determined by the responsible medical student based on reasonable medical judgment. The particular therapeutically effective dose for any particular host may depend on a variety of factors including, for example, the condition to be treated and the severity of the condition; the specific composition of the particular composition used, the particular composition used by the patient , age, weight, general health status, gender and daily diet; time of administration; mode of administration; excretion rate of the particular compound used; duration of treatment; the Quxixi used with or with the particular composition used Compounds and/or such onioncyclin analogs; and similar factors well known in the art of medical technology. Example 97 200831111 It is preferred to administer the composition at a dose lower than that required to achieve the desired therapeutic effect, within the technical scope of the art, and then gradually increase the dosage until the desired therapeutic effect is obtained. In order to facilitate administration and uniformity of the dosage, it is preferred to modulate the composition comprising the triclinide compound and one or more onioncycline analogs in dosage unit form 5. As used herein, &quot;dosage unit form&quot; refers to the actual individual unit of the composition that is suitable for the host to be treated. Each dose should contain itself or associated with a particular pharmaceutical carrier medium and is expected to produce the desired therapeutic effect. The composition of the quantity. 10 15 The parental early dosage formula is such that it contains a daily dose or unit, a daily sub-dose or a suitable fraction of the administered ingredients. For example, about 0.5 mg per day, the compound disclosed in the text can be reduced by a small amount. The volume of solid tumor in the mouse. ^ 丨 can depend on host factors, such as body weight, age, surface area, metabolism, tissue distribution, absorption rate and excretion rate. In one day, two days = human administration. 5 to 7 grams of the two shoulders revealed in the text. Mother's day can selectively give humans about (1) grams of this person. The therapeutically effective dose can depend on many of the best in this item m, then add the agent 'until to obtain the desired therapeutic effect. Included trellidabine compound and sputum ring health can be used together with a sustained release matrix, the matrix:: role to degrade. - Who is going to do the mind to solve When used or refined into human (4), the matrix stores the body fluid for use. For example, the sustained release matrix is selected from biocompatible substances such as liposome, polylactide (polylactic acid), polyethylidene. Lactide (polymer of glycolic acid), polylactide co-glycolide (copolymer of lactic acid and glycolic acid), polyanhydride, poly(raw) vinegar, peptide, hyaluronic acid, collagen, chondroitin sulfate, carboxy Acids, fatty acids, phospholipids, polysaccharides, nucleic acids, polyamino acids, amino acids (such as phenylalanine, tyrosine, isoleucine), polynucleotides, polyvinyl propylene, polyvinylpyrrolidone And polyoxyl. The preferred biodegradable matrix is a matrix of any of polylactide, polyglycolide or polylactide co-glycolide (copolymer of lactic acid and glycolic acid). The ribine compound and one or more onioncycline analogs may also be administered in the form of a liposome. As is known in the art, the liposome is typically derived from a phospholipid or other lipid. The microlipid system is dispersed in an aqueous medium. Single or multi-layer hydrated liquid crystallized Any non-toxic, physiologically acceptable and metabolizable lipid capable of forming a liposome can be used. In addition to one or more of the compositions of the present invention 15, the liposome can contain a stabilizer, a preservative, and a shape Agents, etc. Examples of lipids are natural and synthetic phospholipids and phospholipids choline (lecithin). Methods for forming liposomes are known in the art. The citrifloxacin compound and one or more onion rings The analog can be formulated into an aerosol suitable for administration by inhalation, and is suitable for administration to the respiratory tract. These formulations may be in the form of an aerosol or a solution suitable for use in an atomizer or in a method suitable for infusion. In the form of a fine powder, and may be used alone or in combination with an inert carrier such as lactose. In this case, in one embodiment, the particles of the formulation have a diameter of less than 50 microns, and in another embodiment, the particles The diameter is less than 10 microns. 99 200831111 The composition of the special cililiate compound and one or more onioncycline analogues is used in combination with other (four) substances and/or procedures to treat the above conditions. For example, it is preferred to have a solution, radiation or chemotherapy and one or more of the compositions of the present invention, and then the patient can be administered one of the inventions or 5 10 15 20 of the plurality of compositions called the long micrometastasis. And stabilize, inhibit or reduce the growth of any residual primary tumor. Further Examples Pharmaceutical compositions comprising a triclinic compound and/or a plurality of vegan analogs can be prepared according to known methods for preparing pharmaceutically useful compositions. Formulations are described in many sources that are well known to us and readily available to those skilled in the art. Such as RemingtQn, s

Sciences (Martin EW[1995] Easton Pennsylvania, Mack

Publishing, Company, 19th Edition describes a formulation that can be used with the present invention. Formulations suitable for administration include, for example, aqueous sterile injection solutions which may contain an antioxidant, a buffer, a bacteriostatic agent, and a solute which renders the formulation isotonic with the blood of the intended recipient; and may include suspending agents and thickening Aqueous and non-aqueous sterile suspensions of the agent. Such formulations may be provided in unit doses or in multiple containers (eg, sealed ampoules and vials) and may be stored in a freeze-dried (dry) strip that is only required to be added prior to use, such as water for injection. under. It can be prepared from sterile powders, granules, tablets, etc., and can be used at any time. It should be understood that in addition to the ingredients specifically disclosed above, the present invention may include other agents of the present invention which have been considered in view of the type of the formula. For example, in order to inhibit the growth of cancerous cells, the present invention may be used in combination with at least one additional treatment method including, but not limited to, chemotherapy, radiation therapy, and a therapy for selectively inhibiting Ras carcinogenicity. Or any other treatment known to those skilled in the art of treating and managing cancer, such as the administration of an anticancer agent. 5 API-2 (Quxi Libin) salt can be administered. Examples of pharmaceutically acceptable salts are organic acid addition salts formed using acids which form physiologically acceptable anions, such as tosylate, mesylate, acetate, citrate, malonate, Tartrate, succinate, benzoate, ascorbate, alpha-ketoglutarate, and glycerol phosphate. Suitable 10 inorganic salts can also be formed, including hydrochlorides, sulfates, nitrates, bicarbonates, and carbonates. The pharmaceutically acceptable salts can be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound, such as an amine, with a suitable acid which produces a physiologically acceptable anion. It is also possible to prepare a test metal (e.g., sodium, potassium or lithium) or an alkaline earth metal (e.g., calcium) salt of carboxylic acid 15. The polymorphic ciliben compound and one or more anthracycline analogs may be formulated to be suitable for the selected mode of administration, that is, various methods of intravenous, intramuscular, topical or subcutaneous administration, either orally or parenterally. The pharmaceutical composition of the form is administered to a patient, such as a human or veterinary patient. 20 Accordingly, the tresibine compound and one or more onioncycline analogs of the invention may be systemed with a pharmaceutically acceptable vehicle (ie, a carrier), such as an inert diluent or an absorbable edible carrier. Sexually administered, such as oral. It can be enclosed in hard or soft shell gelatin capsules, can be squeezed into tablets or used directly with the patient's diet. For therapeutic oral administration, such compounds may be incorporated in the form of tablets, buccal, syrup, flakes, and the like. This: raw agent. The composition and the active compound 5 in a specific unit dosage form weight composition using one or more excipients and in the following forms, such as tablets, buccal tablets, capsules, elixirs, suspensions, and the like, and preparations thereof should contain At least the percentage of live preparations may of course vary and preferably may range from about 2 to about 60%. The effective amount of such therapeutically useful ingredients can be obtained. 10 inch servant can contain the following components · 7 doses such as yellow (four), gum arabic, corn coagulum or gelatin: shaped acid diuretic; decomposing agents, such as corn house powder, horse hall powder, 2 acid core lubricant, Such as hard fat Lai Taohe; moon tree oil or cherry seasoning. When the unit dosage form is a capsule, =Τ::Γ' may contain a liquid_, such as a vegetable oil. 5 body units: The material may be present as a coating or may modify the physical form of the solid. For example, _, pills or capsules can be coated with gelatin, 2, = sugar, and the like. The sugar or the granule may contain the present compound, the recommended sugar or fructose as a sweetener, the acid methyl vinegar and the propyl benzoate vinegar, the bismuth (four) taste; the base citrus (four) Α Α wood 1· Ten and unrecognized, such as cherries or 20 content should be two: used to injure any unit of any material ~ I am acceptable and essentially understand that the compounds can be incorporated into sustained release formulations And inside the device. L-external hair can also be administered by infusion or injection. The various kinds of onioncycline analogues are less (4) oral and/or non-toxic intravenous or intraperitoneal administration. The surfactants may be selectively mixed in water to form a solution of the activators or their salts 102 200831111. The village prepares a dispersion in glycerin, liquid polyethylene glycol, triacetin, and mixtures thereof, and in oil. Under normal conditions of storage and use, these (4) contain preservatives that prevent the growth of microorganisms.

Such pharmaceutical dosage forms suitable for injection or bolus injection may include sterile aqueous solutions or dispersions containing the active ingredient suitable for the preparation of a sterile injectable or infusible solution or suspension and optionally encapsulated in the liposome. Liquid or sterile powder. In the case of the cut, the final dosage form must be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle may be a solvent or liquid dispersion medium including, for example, water, ethanol, polyhydric hydric alcohol (eg, glycerol, propylene glycol, liquid polyethylene glycol, etc.), vegetable oil, non-toxic glycerin, And their suitable mixture. The appropriate rhythm + can be maintained by, for example, by forming a micro-relationship, by maintaining the desired particle size (in the case of dispersions) or by using a surfactant. Microbial action can be prevented by various antibacterial agents and antibacterial agents, for example, benzoic acid vinegar, chlorobutanol, sulphuric acid, sorbic acid, and 15 sulphuric acid. In many cases, it may be preferred to include isotonic agents, for example, sugars, buffers, or sodium chloride. An agent capable of delaying absorption, such as mono-hardy acid I and gelatin, may be used in the composition, and the excipient may be called a sputum. The necessary amount of the compound of the present invention and various other ingredients mentioned above may be used. Incorporate the appropriate solution _, = if necessary, follow the (four) line of correcting sputum injection. In the case of avoiding the preparation of non-k ejaculation domain powder, the preparation method is W and the material surface, and the powder is produced in the previous (4) filter (4) material (10), and any material required to be t 2 〇 powder 103 10331111 For topical administration, the trichostatin compound and one or more halogen analogs can be administered in pure form (i.e., liquid). However, they are preferably administered to the skin in a suitable amount and in a composition or formulation of a human acceptable carrier (which may be a solid or liquid). 5 Useful solid carriers include finely divided solids such as talc, point soil, microcrystalline cellulose, ceria, alumina, and the like. Useful liquid carriers include water, alcohol or ethylene glycol or a water-alcohol/ethylene glycol blend wherein the compound of the invention or the like can be dissolved or dispersed in an effective amount by means of a non-toxic surfactant. Adjuvants, such as perfumes and additional antimicrobial agents, may be added to optimize the properties of the particular application. The resulting liquid composition applied from the absorbent pad can be applied to the infected area by dipping the bandage and other dressings or using a pump or aerosol nozzle. Thickeners may also be used in conjunction with the liquid carrier, such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified cellulose or modified minerals 15 to form directly applied to the skin of the user. Can be spread, paste, gel, ointment, soap, etc. Useful dermal compositions which can be used to deliver the compounds of the present invention to the skin are disclosed in the following materials: Jacquet et al. (U.S. Patent No. 4,608,392), Geria (U.S. Patent No. 4,992,478), Smith et al. No. 4,559,157) and 20 Woltzman (U.S. Patent No. 4,820,508). The useful doses of these pharmaceutical compositions of the present invention can be determined by comparing their in vitro and in vivo activities in vivo in an animal model. Extrapolation of effective doses in mice, and other animals to humans is known in the art, see, for example, U.S. Patent No. 4,938,949. 104 200831111 In one non-limiting embodiment, the concentration of the active agent in the liquid composition (such as an emulsion) can range from about 0.1 to 25% by weight or from about 0.5 to 10% by weight. In one embodiment, the concentration in a semi-solid or solid composition (such as a gel or powder) may be from about 0.1 to 5% by weight, preferably from about 0.5 to 2.5, and from about 0.50%. In one embodiment, in the case of an adult, a single dose suitable for injection, infusion or ingestion typically ranges from 5 to 1500 mg and can be administered from 1 to 3 times per day to produce from about 0.1 to 50 mg/kg. dose. A non-limiting oral dose of the present invention may be between 7.5 and 45 mg per day, as appropriate for determining the weight of the individual. 10 Accordingly, the invention includes a pharmaceutical composition comprising a tricitin compound and one or more onioncycline analogs or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier. A pharmaceutical composition suitable for oral, topical or parenteral administration comprising a quantity of a trichostatin compound and one or more onioncycline analogs or pharmaceutically acceptable salts thereof, constitutes a preferred embodiment of the present invention . For the purposes of this invention, the dose administered to a patient, particularly a human, should be sufficient to affect the patient's therapeutic response within a reasonable time frame. Those skilled in the art will appreciate that the dosage will depend on a variety of factors including the health of the animal, the weight of the animal, and the severity of the cancer and the stage of cancer. A suitable dose is one which will allow the concentration of the trichostatin compound and one or more onion 20 cyclin analogs in known tumor tissues to affect the desired response. The preferred dosage is a dose which results in the greatest inhibition of cancer cell growth without uncontrolled side effects. API-2 (or a pharmaceutically acceptable salt thereof) can be administered continuously or at different intervals, as determined by the ordinarily skilled artisan. Nutrients that are useful for inhibiting the growth of cancer cells 105 200831111 Milk species include, but are not limited to, primates such as lynx, sable, orangutan, human, monkey; domesticated animals (eg, pets) Such as dogs, cats, guinea pigs, hamsters, Vietnamese pot-bellied pigs, rabbits, and ferrets; domesticated farm animals such as cattle, buffalo, bison, horses, donkeys, pigs, sheep, and goats; typical 5 in animal rafts Animals found outside the country, such as bears, lions, tigers, leopards, elephants, hippos, rhinos, giraffes, antelopes, sloths, gazelle, zebras, wild bees, prairie dogs, koalas, kangaroos, possums , raccoons, pandas, hyenas, seals, sea lions, elephant seals, otters, porpoises, dolphins, and recorded fish. The terms "patient" and "patient" are used interchangeably herein and are intended to include such human and non-human mammalian species. Likewise, the in vitro method of the invention can be carried out on cells of such mammalian species. Patients who need to be treated using such methods of the invention can be identified using standard techniques known to the medical professional. The following examples are provided for clarification and are not limiting. 15 Example Example 1 : Live It Screening

Cell lines and NCI diversity molecular libraries: All cell lines can be purchased from ATCC or as described above (Cheng, J. Q., et al. Oncogene, 14: 2793-2801, 1997, West, Κ·Α·, et al. Drug Resist · Updat·, 5: 234-248, 2002, 20 Satyamaorthy, K., et al. Cancer Res. 61: 7318-7324, 2001). The NCI structural demultiplexing molecular library is a repository of 1,992 compounds selected from the NCI drug library of about 140, (10) 0 compounds. Further information on the selection, structure, and activity of compounds in these diverse molecular libraries can be found on the NCI Developmental Therapeutics Program website. 106 200831111

Screening for inhibition of Akt-transformed cell growth: NIH3T3 cells transfected with AKT2-transformed NIH3T3 cells or LXSN mediators (Cheng, JQ et al. Oncogene, 14: 2793-2801, 1997) were inoculated into 96 Well tissue culture plate. After 5 μΜ of NCI diversity molecular library compound treatment 5, cell growth can be detected using CellTier 96 One Solution Cell Proliferation kit (Promega), which can inhibit AKT2 transfected cells instead of LXSN metastatic infection NIH3 T3 Compounds of cells are considered to be suitable for Akt inhibitors and subjected to further analysis. Appropriate _ in vitro protein kinase, cell survival and apoptosis assay: In vitro kinase assays are performed as described in the previous 10 (see, for example, Jiang, K· , Coppola, et al.

Mol. Cell·Biol., 20: 139-148, 2000). Cell survival was assayed by MTS (Promega). Apoptosis is detected by annexin v as described above (Jiang, K., Coppola, et al. Mol. Cell. Biol., 20: 139-148, 2000). Recombinant Akt and PDK1 lines were purchased from Upstate 15 Biotechnology Inc. 〇Conclusion Road 1, a small molecule inhibitor of API-2: Akt is frequently detected in human cancer and the destruction of the Akt pathway induces cell death and Inhibition of tumor growth (Jetzt, A., et al. Cancer Res., 63: 697-706, 20 2003). Therefore, Akt is considered an attractive molecular target for the development of novel cancer therapies. To identify small molecule inhibitors (groups) of Akt, the chemical library of 1,992 compounds from the NCI (NCI Diversity Molecular Library) was evaluated to be suitable for inhibiting AKT2 transfected cell growth rather than emptying media. LχSN transfers the agent of infected NIH3T3 cells. Repeated experiments demonstrated that 32 compounds 107 200831111 only inhibited the growth of AKT-2 transformed cells. One of the most potent compounds, An-2 (NCI identification number: NSC 154020), inhibits cell growth at concentrations of 50 Μ. Figure 1 shows ΑΡΙ-2, also known as the chemical structure of triclinide (Schweinsberg, P. D., et al. Biochem Pharmacol, 5 30: 2521-2526, 1981). API-2 is optional. The fact that inhibition of AKT-2 transfected cells, but not untransformed parental cells, helps us determine whether ΑΠ2 is an inhibitor of ΑΚΤ2 sputum. Accordingly, ΑΚΤ2 was immunoprecipitated using an anti-ΑΚΤ2 antibody obtained from ΑΚΤ-2-transformed Τ3Τ3 cells treated with ΑΡΙ-2. The ΑΚΤ2 immunoprecipitate was immunized with an anti-phospho-acidified Akt antibody. As shown in Figure 10B, ΑΠ-2 significantly inhibits AKT2 phosphorylation of threonine-309 and serine-474 required for complete activation of AKT-2 (Datta, SR, et al. Genes Dev 13:2905-2927, 1999). Since the three variants of Akt share high homology and similar structures, the effect of API-2 on their kinase activity can be assessed. The HEK293 cell line was infected with HA-Akt1, HA-AKT2 and 15 HA-AKT3, and the serum was starved overnight and treated with API-2 before stimulation with EGF (50 ng/ml), which took 60 minutes. Three replicate experiments showed that API-2 inhibited EGF-induced kinase activity and squaric acidification of Aktl, AKT2 and AKT3 (Fig. 1C). However, in the in vitro kinase reaction, the kinase activity of recombinant pro-ogenic AKT2 (Myr-AKT2) was not inhibited by API-2 (Fig. 1D), indicating that API-2 did not directly inhibit in vitro Akt and indicates that API-2 is neither a competitor to ATP nor a matrix competitor that can bind to the active site of Akt. API-2 does not inhibit the known upstream activity of Akt ▲ 丨: It has been clearly documented that it can be via P13K-dependent, extracellular stimuli and intracellular 108 200831111 * 5 signaling molecules, such as active Ras and Src, Activate Akt. Thus, the API-2 inhibition of Akt can result from the labeling of upstream molecules (groups) of Akt. Since P13K and PDK1 are direct upstream regulators of Akt (Datta, S.R., et al. Genes Dev. 13:2905-2927, 1999), it is possible to check whether API-2 can inhibit PI3K and/or PDK1. The serum of HEK293 cells was starved and then treated with APL2 or PI3K inhibitor, wortmannin, for 30 minutes prior to EGF stimulation. PI3K was immunoprecipitated with an anti-ρΠΟα antibody. These immunoprecipitates were assayed for in vitro Κ3Κ kinase using pi-4-P • as a matrix. As shown in Fig. 2, the EGF-induced ΡΙ3Κ activity was inhibited by wortmannin 10 and was not inhibited by ΑΡΙ-2. In order to evaluate the effect of ΑΡΙ-2 on PDK1, a method in which the recombinant PDK1 is used to promote the sulphate-309 acidification reaction of ΑΚΤ2 is carried out in the presence of a liposome containing lipofectin. As shown in Figure 2, this assay was strongly inhibited by the control PDK1 inhibitor, staurosporine (IC50 = 5ηΜ). Conversely, at the highest concentration tested (5.1 μΜ) 15, ΑΠ-2 showed only 21% inhibition of the assay. To further assess the effect of ΑΡΙ-2 on PDK1 activation, after treatment of ΗΕΚ293 cells with ΑΡΙ-2, PDK1 was examined for serine 241, which is a residue that is itself phosphorylated and is important for its activity. Autophosphorylation content (Datta, SR, et al. Genes Dev. 13: 2905-2927, 1999). Three replicate experiments demonstrated that the phosphorylation level of 20 PDK1 was not inhibited by API-2 (Fig. 2B). However, the PI3K inhibitor, wortmannin, inhibits EGF-stimulated PDK1 (Fig. 2B). The selectivity of API-2#Akt is much more selective for PKC, PKA, SGK, STAT, JNK, |) 38, and ERK signaling pathways: Akt belongs to the AGC (PKA/PKG/PKC) kinase family, which also includes PKA, PKC, serum - 109 200831111 and adrenal glucocorticoid-inducible kinase (SGK), p90 ribosomal S6 kinase, p70S6K, mitogen- and stress-activated protein kinases and PKC-related kinases . In the AGC kinase family, the protein structures of PKA, PKC and SGK are relatively close to Akt kinase and not other members. Therefore, 5 next, examine the effect of API-2 on the enzymatic activity of these three kinases. HEK393 cells were transfected with HA-labeled PKA, PKCa or SGK. In vitro kinase assays and immunoblot analysis demonstrated that the kinase activities of PKA and PKC alpha were inhibited by guanidine and Ro 31 -8220, respectively, which are PKC inhibitors, whereas API-2 showed no effect on their activity ( Figures 2C and 2E). Furthermore, serum-induced SGK kinase activity is attenuated by wortmannin but not by API-2 (Fig. 2D). In addition, it has been determined whether API-2 has an effect on other carcinogenic survival pathways. Western blot analysis using commercially available anti-phosphorylated antibodies showed that the acidification levels of Stat3, JNK, p38, and Erkl/2 were not affected by API-2 treatment (Figure 2F. These data indicate that API-2 can be 15 Specifically inhibits the Akt signaling pathway. 4PI-2 inhibits cell growth and induces Akt over-regulation, apoptosis in sexual/activated human cancer cell lines: API-2 selectively inhibits the monthly b-force expression of the Akt pathway It should inhibit apoptosis in these tumor cells that proliferate and/or preferentially induce abnormal expression/activation of Akt. Since Akt is often caused by Akt overexpression or pTEN mutation in human malignancies In vivo, API-2 can be used to treat cells that express the structurally active Akt produced by overexpression of AKT2 (OVCAR3, OVCAR8, PNNC1 and AKT2·transformed NIH3T3) or by the PTEN gene The cells of the activating Akt produced by the mutation (PC-3, LNCaP, 110 200831111 MDA-MB-468), and the cells that do not respond to the growth stimulation of IGF-1 (OVCAR5, DU-145, T47D, C0L0357 and LXSN-NIH3T3) and IGF-1 used to activate Akt Melanoma cells that are activated but do not respond to growth stimulation of IGF-1 (Satyamoorthy, K., et al. Cancer Res. 5 61: 7318-7324, 2001). Immunoblotting analysis showed that the phosphorylation of Akt was only Inhibition of Akt or cells that respond to IGF-1 stimulation is inhibited by API-2 (Fig. 3A). Therefore, in Akt-expressing/activated cells, compared to cells with low Akt content, API-2 can inhibit cell growth to a large extent. As shown in Figure 3B, API in Akt-expressing/activated cell lines, 10 LNCaP, PC-3, OVCAR3, OVCA8, PANCn, MDA-MB-468 and WM35 -2 treatment inhibits about 50 to 60% of cell proliferation, but only inhibits about 10 to DU145, OVCAR5, C0L0357, T47D, and WM852 cells that show low levels of Akt or no response to growth stimulation of IGF-1. 20%. Moreover, API-2 can induce apoptosis of cell lines of 8 times, 6 times, 6 times, and 15 3 times, respectively: OVCAR3, OVCAR8, PANC1, and AKT2-NIH3T3. OVCAR5, C0L0357 and LXSN- The API-2 and vehicle (DMSO) treatment of NIH3T3 cells did not reveal significant differences in apoptosis. Therefore, API-2 can Cell growth can be made priority can induce apoptosis in abnormal cells expressing the Akt. 20 ΑΡ_Ι-2 inhibits the downstream palladium of Akt: Akt has been shown to exert its cellular effects via phosphorylation of many proteins (Datta, S.R., et al. Genes Dev. 13:2905-2927, 1999). More than 20 proteins have been identified as Akt matrices, including members of the Forkhead family of proteins (FKHR, AFX and FKHRL1), potato globulin (TSF2), p70S6K, GSK-3 111 200831111 β, p21WAF1/Clpl, p27kipl, MDM2 , Bad, ASK1, ΙΚΚα, etc. Next, examine whether API-2 can inhibit downstream targets of Akt. Since anti-phospho-male saponin, -Bad, AFX' and -GSK-3 a//3 anti-systems are commercially available, the effect of API-2 on their phosphorylation induced by Akt can be determined. 5 rings. After treatment with ΑΡΙ-2 (1 μΜ), OVAR3 cells were lysed and the dots were immunized with respective anti-phosphorylated antibodies. Figure 4 shows that ΑΡΙ-2 can inhibit the degree of phosphorylation of potato globulin to a large extent, which can lead to the stability and upregulation of potato globulin (Dan, HC, et al. J. Biol. Chem., 277:35364- 35370, 2002). Phosphorylation of Bad, GSK-3y5, and AFX was partially attenuated by API-2. These data indicate that API-2 induces cell death and controls cell growth by inhibiting phosphorylation of its downstream targets. Different degrees of API-2 inhibition of Akt downstream targets may be due to the fact that the phosphorylation sites of these targets are also controlled by other kinases (groups), for example, in addition to Akt, ΡΑΚΙ can also be Bad leucine-136 Acidification (Schurmann, 15 A., et al. Mol. Cell Biol., 20: 453-461, 2000). Example 2: Anti-tumor activity of nude mouse tumor xenograft model As previously reported, tumor cells were taken and suspended in PBS and injected subcutaneously into the right flank and left flank of an 8-week-old female nude mouse (2 x 106) Within the cell/flank (Sun, J., Blaskovic, et al. Cancer Res., 59: 4919-4926, 20 1999). When the tumor reached about 100 to 150 mm 3 , the animals were randomly sampled and administered daily with 2 ml of the tromethamine compound and/or the onioncycline analog vehicle by intraperitoneal injection. Control animals received DMSO (20%) vehicle&apos; However, treated animals were injected with a solution of API-2 (1 mg/kg/day) in 20% DMSO. 112 200831111 API-2 inhibits tumors in mice that overexpress Akt. The county has demonstrated frequent overexpression/activation and/or expansion of AKT1 and AKT2 in human ovarian and pancreatic cancers (Cheng, JQ, and Nicosia, SV AKT signal transduction pathway in oncogenesis. In 5 Schwab D? Editor, Encyclopedic Reference of Cancer. Berlin

Heidelberg and New York: Springer; 2001·pp35-7). Inhibition of Akt pathway by PI3K, HSP70, Src and inhibitors of farnesyltransferase controls cell growth and induces apoptosis (Solit, DB) et al. Cancer Res., 63: 2139-2144, 2003, Xu, W., et al. 10 Cancer Res., 63:7777-7784, 2003). Recent studies have shown that intratumoral injection of adrenal adenovirus with dominant negative Akt can also significantly inhibit xenografts with high Akt. Tumor growth (Jetzt, A., et al. Cancer Res., 63: 697_706, 2003). Because API-2 only inhibits Akt signaling and induces apoptosis and cell growth control in cancer cells with high Akt content, Therefore, tumor growth with high Akt content in 15 nude mice is more sensitive to API_2 than tumor growth with low Akt content. According to this, subcutaneous injection of Akt overexpressing cells (OVCAR3, OVCAR8 and PANC-1) Subcutaneously implanted into the right flank of the mouse and implanted these low-Akt-expressing cell lines (OVCAR5 and C0L0357) into the left flank. When the average tumor size is about 1〇〇 to 15〇mm 3 20 , Animals were randomly sampled and taken via vehicle or API-2 (1 mg/kg/day) The intraperitoneal treatment was performed. As illustrated in Figure 4B, the vehicle-treated OVCAR-5 and C0L0357 tumors grew to approximately 800-1,000 mm 3 at 49 days after tumor implantation. At 49 days after tumor implantation, OVCAR3, OVCAR8 and PANC1 tumors treated with vehicle control can grow to about 700-900 mm 3. 113 200831111 API-2 can inhibit tumor growth of 9% by mole of vcaR3, 88% of OVCAR8 and 80% of PANC1, respectively. Conversely, aPI_2 had minimal effect on the growth of OVCAR5 and C0L0357 cells in nude mice (Fig. 4B to 4D and did not show data). API-2 against mouse blood at a dose of 1 mg/kg/day There was no effect on glucose content, body weight, activity and food intake. In the treated tumor samples, Akt activity was inhibited by API-2 and did not change the total Akt content (Fig. 4E). These results indicate that api_2 selectively inhibits the growth of tumors with high Akt content.Example 3: TCN directly inhibits wild-type Akt kinase activity 10 API_2 (TCN) directly inhibits wild-type Akt kinase activity induced by in vitro PDK1 ( Figure 1) This result confirms that API_2 is direct Akt inhibition The basic mechanism may be confirmed and API-2 binding to the PH domain of Akt and / or threonine -308. In vitro kinase assays were performed using PDK1 and Akt in recombinants containing the phospholipidino-inositol-3,4,5-P3 (PIP3), API-2, and kinase buffer 15 as a tissue protein of the matrix. After incubation for 30 minutes, the reactants were separated by SDS-PAGE and exposed to the film. Example 4: TCN can effectively detect cancer-resistant cells in cisplatin, paclitaxel, and tamoxifen-resistant A270CP, C-13, OVCAR433, and MCF7/TAM cells. utility. API-2 overcomes the cisplatin, paclitaxel, and tamoxifen resistance of these cells. The invention has been described with reference to preferred embodiments thereof. Variations and modifications of the present invention will become apparent to those skilled in the art from this disclosure. All such variations and modifications are intended to be included within the scope of the present invention. 114 200831111 • 5 • 10 [Simple description of the diagram 3 Figure 1 shows the identification of API-2 (Queritebine) as a suitable substance for Akt inhibitors from the NCI Diversity Set. Figure 1A illustrates the chemical structure of API-2 (Quxi Libin). Figure 1B illustrates the phosphorylation of AKT2 in AKT2-transformed NIH3T3 cells by API-2. NIH3T3 cells transfected with wild-type AKT2 were treated with ΑΡΙ-2 (1 μΜ), time-consuming, and anti-phospho-Akt-T308 and -S473 antibodies (top group and intermediate group) were used for immunoblot analysis. The bottom group shows the performance of the total AKT2. In Figure 1C, it is shown that API-2 inhibits three variants of Akt. HEK293 cells were transfected with HA-Akt1, -AKT2 and _AKT3 and treated with ΑΡΙ-2 (1 μΜ) or wortmannin (15 pM) before EGF stimulation, so that the cells were lysed and anti-sputum antibody Immunoprecipitation. The immunoprecipitates were subjected to an in vitro kinase assay (top) and immunoblot analysis was performed using an anti-phospho-Akt-T308 (bottom) antibody. The middle group 15 • Shows the expressiveness of Aktl, AKT2 and AKT3 metastasized. Figure 1D illustrates that ΑΠ-2 does not inhibit Akt in vitro. An in vitro kinase assay of the recombinantly active AKT2 recombinant protein in a kinase buffer containing ΙμΜ API-2 (lane 3) was performed. 20 Figure 2 illustrates that API-2 does not inhibit PI3K, PDK1, and closely related members of the AGC kinase family. Figure 2A illustrates an in vitro PI3K kinase assay. Serum deficiency of HEK293 cells and treatment with ΑΙ&gt;Ι-2 (1 μΜ) or wortmannin (15 μΜ) before EGF stimulation took 3 minutes. The cells were lysed and immunoprecipitated with an anti-ΡΙΙΟα antibody. These immunoprecipitates were subjected to an in vitro kinase assay using ΡΙ-4-Ρ as a substrate. The second figure illustrates the effect of ΑΠ-2 on the activation of PDK1 (the top group) in vivo 115 200831111, and the filled circle indicates the inhibition by API-2. The open circle indicates inhibition by the positive control staurosporine, which is a potent PDK1 inhibitor (lC50 = 5 nM). The bottom panel was an immunoblot analysis of HEK293 cells transfected with Myc-PDK4 and treated with wortmannin or API-2 prior to EGF stimulation. The immunoblots are detected by the designated antibodies. Figure 2C depicts immunoblotting of PKCa phosphorylation using anti-phospho PKCa T638 (top) and assembly PKCa (bottom) antibodies after treatment with API-2 or a non-selective PKC inhibitor R〇31-8220 Point analysis. Figure 2D shows the SGK kinase assay in vivo 10 . HEK293 cells were transfected with HA-SGK and treated with API-2 or wortmannin prior to EGF stimulation. In vitro kinase assays were performed using HAP as a substrate (top) with HA-SGK immunoprecipitates. The bottom panel represents the expressiveness of HA-SGK transfected with infection. Figure 2E illustrates the results of the PKA kinase assay. The immunopurified PKA was incubated in ADB buffer (Upstate Biotechnology Inc) containing the indicated inhibitor (API-2 or AKAI) and the conjugated substance, Kemptide. The kinase activity was quantified. The 2F figure represents the Western dot method. Treatment of 0VCAR3 cells with ΑΠ-2 took time to specify. Immunoblots of cell lysates were performed with the indicated anti-phospho antibodies (Groups 1 to 4) and anti-actin antibodies (bottom). 20 Figure 3 shows that ΑΠ-2 inhibits Akt activity and cell growth and induces apoptosis in human cancer cells with high Akt: Figure 3A shows Western blotting, after treatment with ΑΠ-2, in the designation The degree of phosphorylation of Akt was detected by anti-phospho-Akt-T308 antibody in human cancer cell lines. These dot methods were then probed with anti-total Akt antibodies (the bottom panel). Figure 3B shows cell proliferation 116 200831111 5 • Verification. The cell lines shown in the figure were treated with different doses of API-2, which took 24 hours and 48 hours, and then analyzed by the CellTiter 96 Cell Proliferation Assay Kit (Promega). Figure 3C provides a cell > weekly analysis. Cells were treated with API-2 and stained with annexin V and PI, and then analyzed by FACScan. Figure 4 shows that API-2 inhibits downstream targets of Akt and displays anti-tumor organisms in cancer cell lines with high Akt in mouse xenografts. Figure 4A shows that API-2 inhibits the Akt phosphatization of potatorin, Bad, AFX and GSK-3/5. After treatment with API-2, OVAR3 cells were lysed and the ink spots were immunized with the indicated antibodies. Figure 4B shows that A PI-2 inhibits tumor growth. Tumor cells were injected subcutaneously into nude mice with low Akt cell content on the left and high Akt cell content on the right. When the tumors reached an average size of about 1 to 150 mm 3 , the animals were treated with vehicle or 1 mg/kg/day of API-2. Each measured value represents the average of 10 tumors. Figure 4C is a graphical representation of mice treated with API-2 or vehicle (control) with VCAR3 (right) and 0VcaR5 (left) 15 • xenografts. Fig. 4D shows the tumor size (bottom) and weight (top) at the end of the experiment. In Figure 4E, anti-squamous Akt_S473 (top) and anti-AKT2 (lower) antibodies were used for tumor lysis in treated (T3 and T4) and untreated (T1 and T2) OVCAR-3-derived tumors. Immunoblot analysis of the product. 20 Figure 5 shows that API_2 (Cucidine) inhibits in vitro kinase activity. In vitro kinase assays were performed in recombinant kinases containing PDK1 and Akt in a kinase buffer containing phospholipid creatinine-3,4,5-P3 (PIP3), API-2 and tissue protein as matrix. After 30 minutes of incubation, the aliquots were separated by SDS_PAGE and exposed to the film. 117 200831111 Figures 6a to c provide mRNA and amino acid sequences of human Aktl, and also record restriction enzyme positions. Figures 7a to d provide human Akt2 and mRNA amino acid sequences, as well as restriction enzyme positions. 5 Figures 8a to c provide mRNA and amino acid sequences of human Akt3, as well as restriction enzyme positions. [Main component symbol description] (none) 118

Claims (1)

200831111 X. Patent Application Range: 1. A composition comprising: (i) a compound of the formula I to IV or a salt thereof: Wherein R 2 ', R 3 ' and R 5 ' are each independently hydrogen, optionally substituted phosphate or phosphonate (which includes mono-, di- or triphosphate or stabilized phosphate prodrugs); It includes a low carbon fluorenyl group; an alkyl group (which includes a low carbon alkyl group); a decylamine, a benzoic acid, which includes a decyl or an aryl group; a sulfonyl group, 119 10 , a fluorene group And a benzyl group, wherein the phenyl group may be optionally substituted with one or more substituents such as those described in the definition of aryl groups as described herein, optionally substituted sulfhydryl groups; lipids, including disks Lipid; 2 base acid; carbohydrate; peptide; or cholesterol; or other pharmaceutics in which two feet, 2, R3 and R5' are independently H or mono-, di- or tri-phosphate. Acceptable de-bonding group; long wherein RX&amp;Ry is independently hydrogen, optionally substituted phosphate; $yl (which includes lower sulfhydryl); decylamine, alkyl (which includes lower alkyl); a polyalkylene oxide, such as polyethylene glycol, a selectively substituted * group, which includes a lipid; an amino acid; a carbohydrate; a peptide, Cholesterol; or other pharmaceutically acceptable leaving group. In a reality, the 5H compound is administered as a 5'-acid-lipid or lipid; Ri and R2 are each independently a linear, optionally substituted linear, branched or cyclic alkyl group (including Low alkyl, alkenyl or alkynyl, 〇〇-alkyl, C0-alkenyl, C0-fast, C0-aryl or heteroaryl, c oxyalkyl, C0-arylalkyl a C0-substituted aryl group, a fluorenyl group, a calcined zeolitic group, an aromatic sulfonyl alcohol group, an aromatic fluorinated fluorenyl group; (ii) a compound of the formula V or a formula VI, wherein the compound of the formula VI has the following structure ·· 120 200831111 Formula V Ri, R4, R6, and R7 are each independently nitrogen, a trans group, and an alkoxy group; r2 and each independently are hydrogen, an alkyl group, an alkoxy group, a hydroxyl group, or a halogen; and r5 is a hydrogen, optionally substituted alkyl chain. a branched or cyclic alkyl group, a hydroxyl group, an alkoxy group; R8 is a CO-alkyl group, a CO-halogen-substituted alkyl group, a CO-aryl group or a heteroaryl group; and wherein the compound of the formula VI has the following structure: 1 and 112 are each independently hydrogen, alkyl, alkoxy, hydroxy, halogen; R3 and R4 are each independently hydrogen, optionally substituted alkyl, branched or cyclic alkyl, optionally a substituted alkyl chain, a branched chain or a cyclic alcohol, an optionally substituted alkyl chain, a branched chain or a cyclic amine; and (iii) a pharmaceutically acceptable carrier. 121 15 200831111 2. The composition of claim 1, wherein the compound of formula I is triclinide. 3. The composition of claim 1, wherein the compound of formula I is triclinide phosphate. 4. The composition of claim 1, wherein the compound of formula I is triclinide. 5. The composition of claim 1, wherein the onioncyclin analog is a compound of formula V: 10 Ri, R 4 , R 6 , R 7 are each independently a benzyl or alkoxy group; R 2 and R 3 are each independently hydrogen, alkyl, alkoxy, hydroxy, halogen; R 5 is alkoxy; and R 8 is CO-alkyl , CO-halogen substituted alkyl, CO-aryl or heteroaryl. 6. The composition of claim 1, wherein the onioncyclin analog is a compound of formula VI: 122 200831111 1 and 112 are each independently an anthracene, a decyl group, an alkoxy group, a transatom group, a halogen; and R3 and R4 are each independently hydrogen, a optionally substituted alkyl chain, a branched chain or a cyclic alkyl group, optionally A substituted alkyl chain, a branched chain or a cyclic alcohol, an optionally substituted alkyl chain, a branched chain or a cyclic amine. 7. The composition of claim 1, wherein the onioncyclin analog is a compound selected from the group consisting of: 10 A Idarubicin B Daunorubicin C Cranberry (Doxorubicin) D Epibicin E Pirarubicin F Vaimbicin 123 200831111 M G Mitoxantrone 8. The composition of claim 1, wherein the compounds of formula I to IV are present in a dose of at least 10 mg/m2. 5 9. The composition of claim 1 wherein the compound of formula V to VI It is present in an amount of at least 1 mg. 10. The composition of claim 1 is suitable for parenteral administration. 11. The composition of claim 10, wherein the parenteral administration is intravenous administration. 10 12. The composition of claim 1 of the patent application, which is suitable for oral administration. 13. The composition of claim 1 is suitable for topical administration. 14. A method of treating a tumor or cancer in a mammal comprising administering to the mammal an effective amount of a composition comprising: (1) Compounds of the formula: 124 200831111 Wherein R 2 , R 3 ' and R 5 ' are each independently hydrogen, optionally substituted a sulphate or phosphonate (including mono-, di- or tri-squarate or diazepam 5 acid prodrugs) ); a base (which includes a low-carbon base); a base (which includes a low-carbon base); a sulphate, a sulphuric acid, a sulphur-based or a sulphur-based base, a continuation base, which includes methanesulfonate And a phenyl group, wherein the phenyl group is optionally substituted with one or more substituents as described, for example, in the definition of an aryl group as taught herein; a selectively substituted arylsulfonyl group; a lipid, Including phospholipids; amino acids; carbohydrates; peptides; or cholesterol; or other pharmaceutically active compounds in which R2, R3, and 1, which are independently hydrazine or mono-, di- or tri-phosphate Acceptable leaving group; wherein Rx and Ry are independently hydrogen, optionally substituted sulphate; fluorenyl (which includes lower sulfhydryl); decylamine, alkyl (which includes lower alkyl); Aromatic private polyalkylene oxides, such as polyethylene glycol, selectively substituted aromatic stones, lipids, including phospholipids; amino acids; Carbohydrate; a peptide; or cholesterol; or other pharmaceutically acceptable leaving group. In Yi Shi 125 200831111, the compound is administered as a 5,-phosphate ether lipid or a 5,-ether lipid; Ri and R2 are each independently a linear, optionally substituted straight chain, branched chain or ring system. Alkyl (which includes lower alkyl), alkenyl or alkynyl, CO-alkyl, CO-alkenyl, CO-alkynyl, CO-aryl or heteroaryl, C0-alkoxyalkyl, CO- An aryloxyalkyl group, a CO-substituted aryl group, a sulfonyl group, an alkanesulfonyl group, an arylsulfonate group, an arylsulfonyl group; (ii) an indium cycline analog of the formula V or a formula VI, wherein the compound of the formula I Has the following structure: 10 Formula V Ri, R4, R6, each independently hydrogen, hydroxy, alkoxy; R2 and R3 are each independently hydrogen, alkyl, alkoxy, hydroxy, halogen; R5 is hydrogen, optionally substituted alkane Base group, branched chain or ring system alkyl, hydroxy, alkoxy; 15 R8 is CO-alkyl, CO-halogen substituted alkyl, CO-aryl or heteroaryl; : 126 200831111 1 and 1^2 are each independently hydrogen, alkyl, alkoxy, hydroxy, halogen; R3 and R4 are each independently hydrogen, optionally substituted alkyl chain, branched 5-bond or ring-based alkyl, optionally A substituted alkyl group, a branched bond or a cyclodecyl alcohol, an optionally substituted alkyl chain, a branched chain or a cyclic amine. The method of claim 14, wherein the compounds of the formulae I to IV and the compounds of the formulae V to VI are administered simultaneously. 16. The method of claim 14, wherein the compound is administered to the compound of formula V to VI. 17. The method of claim 14, wherein the compound of formula I to IV is administered once a week for 3 weeks, and then the compound is not administered over a period of one week. 18. The method of claim 14, wherein the compound of formula V to VI is administered once a week for 3 weeks, and then the compound is not administered for a period of one week. 19. The method of claim 14, wherein the compound of formula V is administered once per week for 3 weeks, and then the compound is not administered during a week. 2〇 2〇· The method of claim 14, wherein the dosing schedule is repeated at least twice. 127 200831111 21. The method of claim 14, wherein the dosing schedule is repeated at least four times. 22. The method of claim 14, wherein the treated tumor is a breast, pancreas, ovary, and colorectal tumor. 5. The method of claim 14, wherein the compound of formulas I to IV is administered at least 10 mg/m2. 24. The method of claim 14, wherein the compound of formulas I to IV is administered at a dose of 10 mg/m2 or less. 25. The method of claim 14, wherein at least 1 mg of the compound of formula V to VI is administered. 26. The method of claim 14, wherein at least 10 mg or less of the compound of formula V to VI is administered. 128