TW200902034A - Effective treatment of tumors and cancer with prodrugs of triciribine - Google Patents

Abstract

This application encompasses novel prodrugs of triciribine and related compounds and therapeutic regimens of prodrugs of triciribine and related compounds and compositions for the treatment of tumors, cancer, and other disorders associated with abnormal cell proliferation. The application further encompasses oral formulations of prodrugs with increased bioavailabilty, which allows a new route of administration with the attendant clinical benefits of ease of administration.

Description

。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 5 1. FIELD OF THE INVENTION The present invention encompasses novel triciribine prodrugs and related compounds, as well as triclinib prodrugs and related compounds and compositions for the treatment of tumors, cancer and other disorders associated with abnormal cell proliferation. Treatment plan. The case further covers increased bioavailability, allowing new routes of administration 10 to be accompanied by an oral formulation that is easy to administer. C Prior Art 3 2. Background of the Invention Cancer is an abnormal growth of cells. Despite being limited by space, sharing nutrients with other cells, or signaling from the body to stop replication, cancer cells are still rapidly replicating. Cancer cells often have different shapes from normal healthy cells, fail to function properly, and may spread to multiple areas of the body. Abnormal growth of tissue is referred to as a tumor, which is a cluster of cells that can be uncontrollable for growth and division. The tumor may be a benign (non-cancerous) tumor or a malignant (cancerous) tumor. Benign tumors grow slowly and do not spread. Malignant tumors can grow, invade and destroy adjacent abnormal tissues at a rapid rate and spread throughout the body. Cancer can be classified according to the type of body fluid or tissue from which it is derived, or it can be classified according to the site in which the cancer first appears in the body. In addition, several cancers are mixed. Cancer can be divided into five broad categories, namely cancer, sarcoma, lymphoma, leukemia, and myeloma, indicating tissue classification and blood classification of cancer. 5 200902034 Carcinoma is a cancer called epithelial tissue that appears in body tissues. The epithelial tissue is covered or padded on the surface of organs, glands or body structures. For example, a stomach-lined cancer is referred to as a cancer. A variety of cancerous effects involve the secretion of organs or glands, such as the mammary glands that make milk. Carcinoma accounts for 80% to 90% of all cancer cases. Sarcomas are malignant tumors that are grown by connective tissues such as cartilage, fat, muscle, tendons, and hard bones. The most common sarcoma is a tumor that grows on a hard bone and is common in young adults. Examples of sarcomas include osteosarcoma (hard bone) and soft osteosarcoma (cartilage). Lymphoma refers to a cancer derived from the lymph nodes or lymph nodes of the lymphatic system, which is responsible for the production of white blood cells and cleansing body fluids, or lymphomas derived from organs such as the brain and breast. Lymphomas are divided into two broad categories: Hodgkin's lymphoma and non-Hodgkin's lymphoma. Leukemia, also known as blood cancer, is a bone marrow cancer that causes the bone marrow to fail to produce normal red blood cells and white blood cells and platelets. White blood cells are needed to fight infection. Red blood cells are needed to prevent anemia. Platelets keep the body from dark and 15 blood. Examples of white jk diseases include acute myeloid leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia, and chronic lymphocytic leukemia. The terms bone marrow and lymph are the source of the cells involved. Finally, myeloma is caused by the growth of plasma cells in the bone marrow. In some cases, myeloma cells assemble in a hard bone to form a single tumor, called a plasmacytoma. However, in the other 20 cases, myeloma cells were aggregated into multiple hard bones to form multiple bone tumors. Called as multiple myeloma. Tumor induction and tumor progression are often the result of cumulative changes in the tumor cell genome. Such changes may include deactivation of a cell growth inhibitory gene or tumor repressor gene, as well as activation of the cell growth promoting gene 200902034 or an oncogene. Hundreds of activated cellular oncogenes have been identified in animal models, but only a few genes have been shown to be associated with human cancer (Weinberg et al., the genes of oncogenes and cancer, 1989, Cold Spring Harbor Laboratory, New York; Stanbridge j et al. 丨99〇, ' 5 63: P 867-874; Godwin et al., 1992 'Oncogenes of gynecological malignancies' and anti-oncogenes. In H〇skins WJ·, Perez CA and Young RC (ed.), Gynecologic Oncology: Principles and Practice, pp 87_116, Lippincott, Philadelphia). The activation of oncogenes in human cancer may come from factors such as an increase in the number of gene copies or structural changes. These factors cause a number of cellular effects, such as these factors may lead to excessive expression of the gene product. Several oncogenes involved in human cancer can be activated by gene overexpression. It is clear that the continuous gene disorder obtained by the cancer gene leads to the regulation of normal cell proliferation, differentiation and regulation of apoptosis in the regulation of signal transduction 15 loop defects (Hanahan, D. and Weinberg R_A., Cell, 2000. 100 (1): p. 57-700). This in turn leads to basic defects in cell physiology, resulting in malignant diseases. These defects include: a) self-deficiency in growth signals (ie, excessive expression of growth factor receptors such as EGFR, and downstream signal transduction pathways such as Ras/Raf/Mek/Erk 1/2 and Ras/PI3K/Akt disorder 20 activation) 'b) resistance to growth signals (ie, decreased expression of TGFp and its receptors), c) avoidance of apoptosis (ie loss of pre-apoptotic p53; Overexpression of surviving Bcl-2; survival pathways such as overactivation of the survival pathway mediated by pI3K/Akt), d) persistent angiogenesis (ie, high secretion of VEGF); and e) tissue invasion and metastasis (also That is, extracellular protease and pre-transfer junction 7 200902034 (Hanahan, D. and Weinberg R_A_, Cell, 2000. 100(1): p. 57-700). Receptor tyrosine kinases such as EGFR, ErbB2, VEGFR, and the insulin series growth factor I receptor (IGF-1R) are closely involved in a variety of human cancers, including colorectal cancer, pancreatic cancer, breast cancer, and ovarian cancer (Khaleghpour). Κ· et al., Cancer, 2004.25(2); p.241-8; Sekharam Μ. et al., Cancer Research, 2003, 63(22): ρ_7708-16). Ligands such as EGF, VEGF, and IGF-1 bind to their receptors, contribute to the stimulation of specific tyrosine kinase activity, autophosphorylation of specific tyrosine at the cytoplasmic functional site of the receptor, and trigger a variety of complexities. Signaling of signal transduction pathways (Olayioye Μ·Α. et al, Embo J, 2000. 19(13): p. 3159-67; Porter AC and Vaillancourt RR, oncogenes, 1998.17 (11 ): p.1343-52). This in turn leads to a variety of tumor survival pathways and tumor pathologies such as Ras/Raf/Mek/Erk 1/2, JAK/STAT3 and PI3K/Akt pathways. Although all three pathways are implicated in tumorigenesis of colorectal cancer, pancreatic cancer, breast cancer, and ovarian cancer, the path of Akt media has been shown to be critical in multiple steps of malignant transformation, malignant transformation. The steps include cell proliferation, anti-apoptosis/survival, invasion and metastasis, and tumor neoplasia (Datta S_R. et al., Gene Dev, 1999. 13(22): p_2905-27). 2〇 Akt is a serine/threonine protein kinase (also known as PKB), and Akt has three family members Aktl, Akt2 and Akt3. Stimulation of cells with growth factors or survival factors results in recruitment to the receptor for phosphokinase phosphoinositide-3-〇11-kinase (?13). 13 Phosphorylation of phosphoinositide-4,5-diphosphate (PIP2) into PIP3, PIP3 recruits Akt to the plasma membrane, in the plasma membrane, 200902034

Akt is activated by phosphorylation of Thr308 and Ser473 (Aktl), Thr308 and Ser474 (Akt2) and Thr308 and Ser472 (Akt3) (Datta S.R. et al., Gene Dev, 1999.13(22): ρ·2卯5-27). Thus, PI3K activates Akt by phosphorylating PIP2 and converting PIP2 to PIP3. The phosphatase PTEN 5 dephosphorylates PIP3 to PIP2, thus preventing the activation of Akt. Most human cancers contain over-activated Akt (Datta SR et al., Gene Dev, 1999. 13(22): p. 2905-27; Bellacosa A. et al., International Journal of Cancer, 1995. 64(4): ρ·280- 5; Sun M. et al., Am J Pathol, 2001·159(2): p. 431-7). In particular, Akt was overexpressed and/or highly activated in 57%, 32%, 10 27%, and 36% of human colorectal cancer, pancreatic cancer, breast cancer, and ovarian cancer, respectively (Roy HK et al., Cancer Occurrence, 2002. 23(1): p.201-5; Altomare DA et al., J Cell Alienation. 116111, 2003· 88(1): p. 470-6; Sun M. et al., Cancer Research, 2001_ 61 (16 ): p.5985-91; Stal 0. et al., Breast Cancer Research, 2003.5(2): ρ·R37-44; 15 Cheng JQ· et al, Proc Natl Acad Sci USA, 1992. 89(19): p. 9267-71; Yuan ZQ· et al., Oncogene, 2000. 19(19): p. 2324-30). Overactivation of Akt is due to amplification and/or overexpression of Akt itself, as well as genetic changes upstream of Akt, including overexpression of receptor tyrosine kinase and/or its ligands (Khaleghpour K et al., Cancerogenesis, 2004. 20 25(2); p.241-8; Sekharam M. et al., Cancer Research, 2003. 63(22): p.7708-16; Cohen B_D. et al., Biochem Soc Symp, 1998. 63: P.199-210; Muller W.J_ et al., Biochem Soc Symp, 1998. 63: p-149-57; Miller W. E_ et al, J Virol, 1995. 69(7): p.4390- 8; Slamon D. J. et al., Science, 1987· 235 (4785): p_177-82; Andrulis 9 200902034 1丄· et al, J Clin Oncol, 1998. 16(4): p.1340-9), and Acid _ PTEN's censored. The concept of Akt involved in tumorigenesis confirms that it has been validated in the preclinical phase, showing that ectopic manifestations of Akt induce malignant transformation and contribute to cell survival (Sun M. et al, Am J Pathol, 2001. 159(2): ρ·431 -7; 5 Cheng JQ et al., Oncogene, 1997. 14(23): ρ·2793-801); and the destruction of the Akt pathway inhibits cell growth and induces apoptosis (Jetzt A. et al., Cancer Research, 2003.63 (20): ρ·6697-706). Current treatments for cancer and related diseases have limited efficacy and a variety of serious undesired side effects. Although the clinical efficacy of a variety of anticancer drugs has been validated, its severe systemic toxicity often results in clinical disruption of a variety of promising chemotherapeutic agents. Furthermore, overexpression of receptor tyrosine kinases such as EGFR and its ligands such as IGF-1, overexpression of Akt and/or loss of PTEN (all leading to excessive activation of Akt) result in poor prognosis and resistance to chemotherapy. The medicinal properties and the survival time of cancer patients are shortened. The current research strategy line 15 emphasizes the study of effective treatment modalities with lower risks. 2.1. Quxi $ Quxilide (TCN, NSC-154202, 3-amino-1,-dihydro-5-mercapto-l-β-ribosefuranosyl-1,4,5,6,8 - Wuxi) and its 5'-phosphate, the anticancer effect of Tricinel (TCN-P, NSC-280594) was first identified in the 1970s (Townsend & Miline (1975) Ann NY Acad Sci, 255:92-103). TCN-P becomes a chemical entity that progresses into clinical trials due to the higher solubility of the parental drug. By the early 1980s, TCN-P had been shown to be anti-leukemia and carcinoma and preclinical activity. By the early 1980s, TCN_p had been identified as an inhibitor of DNA, RNA, and protein 10 200902034 synthesis, and TCN-Ρ was selective for cell selection during the s phase of the cell cycle (R〇ti-R〇ti et al' 1978 Proc Am Assoc Cancer Res and ASCO 19:40). It also suggests that unlike other nucleoside antitumor agents at the time, TCN-P is not acidified by the disc to the extent of one disc acid salt and does not bind to the polymer.

5 Acid glycosides (Bennett et al. 1978 Biochem Pharmacol 27:233-241, Plagemann JNCI 1976 57:1283-95). Established in vivo, TCN-P is dephosphorylated to TCN by plasma enzyme and by extracellular-5'-nucleotidase. Within the cell, TCN can be phosphorylated again to TCN-P by adenosine kinase (Wotring et al, 1981 Proc. Am Assoc Cancer Ress 10 22:257, Basseches et al, J Chromatogr 1982 233:227-234). In 1982, TCN-P entered Phase I clinical trials by Mittelman and colleagues for 20 patients with progressive, malignant malignancies (Mittelman et al., 1983, Cancer Therapy, Rep 67: 159-162). . TCN-P was administered by intravenous (i.v.) infusion at a dose of 25 mg/m2 to 350 mg/m2 every 15 weeks. The patient used in the experiment was diagnosed with breast cancer, head and neck cancer, lung cancer, pancreatic cancer, squamous cell carcinoma, melanoma or unspecified cancer. It was found that there was only a limited therapeutic response and the toxicity was significant. Mittelman's group found that the use of its dosing did not guarantee further clinical trials, but provoked other research groups to examine the efficacy of TCN-P 20 for certain cancer categories. In 1983, Lu et al. (ASCO Abstracts, Clinical Pharmacology, 34 C-133) also examined the clinical practice of TCN-P on a 5-day visit to a patient via a continuous infusion of 30-40 mg/m 2 via a venous catheter. Lu et al. reported that TCN contributes to hepatotoxicity and anemia, suggesting that these toxicities should be monitored. 11 200902034

Cobb et al. (Cancer Treatment Report 1983 67: 173) reported the activity of surgical explants of TCN-P against human tumors in a 6 曰 suprarenal capsule assay. A total of 80 tumor types were examined for breast cancer, lung cancer, colorectal cancer, ovarian cancer, and cervical cancer. Cobb et al. reported a responsive response rate of 5 different tumor TCNs ranging from 21% (breast cancer) to 88% (cervical cancer).

Feun et al. reported another phase I clinical trial in 1984 (Cancer Research 44 (8) 3608-12). Feun et al. administered a continuous intravenous infusion of 10, 20, 30, and/or 40 mg/m2 for 5 days every 3 to 4 weeks or every 6 weeks. The patient in the experiment 10 was diagnosed with colorectal cancer, sarcoma, melanoma, lung cancer or "other" cancer. Feun et al. reported significant toxicity observed, including hyperglycemia, hepatotoxicity, and thrombocytopenia. The authors recommend a daily sputum experimental program of 20 mg/m2 for 5 weeks. The author also recommends that patients should be closely monitored for liver function and pancreatic function due to toxicity, and must have 15 diabetes. 'A patient with abnormal liver function or a large number of liver metastases. In 1986, Schilcher et al. (Cancer Research 1986 46: 3147-3151) reported the results of a weekly evaluation of TCN-Ρ using weekly intravenous injections. A total of 24 patients with degenerative solid tumors were enrolled in the 42-day study period on the 1st, 8th, 15th, and 22th, with a slow intravenous injection of 2 〇 for 5 minutes, and rested for two weeks during each week. A total of 106 doses were administered from 3 to 12 patients at each concentration to study a concentration of five doses of 12 mg/m2 to 96 mg/m2. The patient used in the experiment has been diagnosed with colorectal cancer, rectal cancer, cancer, adrenal cancer, nest cancer, pancreatic cancer, sarcoma, melanoma, lung cancer or "other" cancer. Schilcher et al. concluded that “this weekly 12 200902034 dosing program produces unpredictable clinical toxicity that cannot be used.” “This study did not use the weekly or intermittent dosing program to give TCN-Ρ for further study. If TCN-P is significantly active against test tube activity against primary pancreatic tumors and liver tumors that are resistant to treatment, then the use of non-administered dosing (eg, monthly-single application) will be resumed. Phase I-II Study". In 1986, P〇wis et al. (Cancer Treatment Report 7: 359_362) reported the distribution of TCN-P in the blood and plasma of patients during Phase I and Phase I clinical trials. The first phase of the experiment used a dose of 24-55 mg/m2 for 10 cycles, while the second phase of the clinical trial used a single dose of 25 mg/m2. Powis et al. did not recognize the interaction between TCN-P pharmacokinetic parameters and TCN_p toxicity. In the late 1980s and early 1990s, TCN_p progressed to the first stage of metastatic colorectal adenocarcinoma, non-small cell lung cancer, cervical squamous cell carcinoma of the cervix, and metastatic breast cancer. In 1987, 〇, c〇nndi et al. (Cancer Treatment Report 7 Ng 3, 333_34) disclose the results of Phase II clinical trials in patients with metastatic colorectal adenocarcinoma. Patients are given a three-week interval once a week for 15 minutes. Intravenous administration of 165 mg / m 2 or 250 mg square TCN P. 〇c_el ^ people obtained conclusions experiments show that Cui 20 for the treatment of patients with metastatic colorectal adenocarcinoma lack of clinical use.丨 991, Lyss et al. (Ρπκ: ΑMeetAmS〇c

Clm 0, (1996) 15 Α 115ι) reported preliminary results of a 5-day daily dose of 35 mg/m2 for patients with exacerbated non-small cell lung cancer. 13 200902034

Feun et al. (Am J Clin Oncol 1993 16:506-508) reported the results of Phase II trials using TCN-P in patients with progressive cervical squamous cell carcinoma. Continuous infusion of at least 35 mg/m2 for 5 consecutive days every 6 weeks. Of the 21 evaluable patients, only two responded. Feun et al. obtained the theory of the use of TCN-P for metastatic or recurrent cervical squamous cell carcinoma with limited activity. In 1996, Hoffman et al. (Cancer Chemotherapy Pharmacology 37:254) -258) Reported the results of the ΐ-π phase study of TCN-P for metastatic breast cancer. In one study, 14 patients were given a continuous intravenous infusion of 2 〇 10 g / m 2 / calendar for 5 weeks every 6 weeks. The treatment was accepted. When the authors failed to observe the response at this dose, the same 5-day continuous infusion was used but the dose was increased to at least 35 mg/m2. Hoffman et al. concluded that "TCNk is not valid for all test doses," And an unacceptable toxicity at doses greater than or equal to 35 mg/m2." 15 Thus, limited efficacy and toxicity are not acceptable for both, impeding further clinical development of TCN-P and related compounds. The 核3/〇79748 issued to the University of California Board of Directors reveals combinations of certain ZNF217 inhibitors such as tricineribine with additional chemotherapeutic agents such as doxorubicon. The 20 I invention encompasses new pharmaceutical compounds and triclinide prodrugs and compositions for the treatment of tumors, cancer, and other conditions associated with abnormal cell proliferation.

t 明内; J 3. SUMMARY OF THE INVENTION 14 200902034 The present invention encompasses the pre-drugidine prodrug that can be used to modulate Akt kinase activity. In particular, chemical agents that can be used to modulate the activity of Akt kinase include, but are not limited to, compositions and formulations comprising the pre-drugidine. 10 15 20 The present invention encompasses novel therapeutic agents for the administration of tromethine, triclinide phosphorylate and related compounds to treat or prevent cancer or cancer, while limiting systemic toxicity. The present invention is based on the discovery that tumors or cancers that overexpress Akt kinase are particularly susceptible to the cytotoxic effects of the triclinic prodrugs and related compounds. Contrary to prior art and experience, the inventors have determined how to successfully use uranium, travertine and related compounds, to treat tumors and cancer in one or a combination of the following: (1) One or more pre-drugs of tricineridine, triclinide and related compounds are administered to patients with increased sensitivity to the drug; (ii) the prescribed dose level is used to minimize the toxicity of the drug While still having efficacy; or (out) using the dosing schedule, which minimizes the toxicity of the drug. The present invention further encompasses the treatment of tumors and cancers that are particularly sensitive to the toxic effects of triclinbine, triflurexine phosphate, and related compounds, which encompasses the treatment of mammals = tumors containing The mammal in need is administered with one or more kinds of triclinide, sputum sirolimus and related compounds. In another embodiment of the invention, the administration of the drug regimen limits the side effects of the tromethamine prodrug, TCN and related compounds. Such a dosing plan can reduce or eliminate toxic side effects, including but not limited to hepatotoxicity and platelet degeneration:

Sputum, hypocalcemia, anemia, hypoalbuminemia · A 呵 · inhibition, blood triacid 15 200902034 glycerin vinegar too high, A transfer plum too high, simple, gastritis and / or fever. Administration of &lt;RTI ID=0.0&gt;&gt;&gt;&gt;&lt;/RTI&gt; </RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; 5 10 15 20 In another embodiment, a method is provided for treating an individual who has been diagnosed with a tumor by administering to the individual an effective amount of one or more Tricinem via a dosing schedule The drug is pre-druged with trulinidin and related compounds, which includes administration of the drug about once a week for about 3 weeks, followed by one week of not administering the drug. In another embodiment, a method for treating a tumor or cancer in a body is provided, by administering one or more of _g/m2 or less per week to the decoction administration plan. Precursor of tromethine, triterpene phosphate and related compounds. In one embodiment, the prodrug compound can be administered in a single-dose dose for a short period of time, e.g., about 5, for 15 minutes. In the case of % sputum, a dosing schedule is provided wherein the prodrug compound is administered through a continuous infusion for at least 24, 48, 72, 96 or 12 hours. In several embodiments, the continuous administration can be repeated at least once a week, every four times, and/or once a month. In other embodiments, the second drive compound may be at least better than the Lin embodiment every three weeks, and the prodrug compound may be administered at least once a day for at least 2, 3, 4, or 5: in additional embodiments As disclosed herein, one or more of the pre-drugs of tromethamine, troxtiltine and related compounds can effectively cause swelling, and the number of drugs administered to the patient. The administration of one or more of the trichostatin, the acid koxi 16 200902034 5 1〇15 2 〇 滨 and related compounds may provide at least partial reaction in at least 15%-20% of the individual in vivo, such as at least 15%, 2〇% or 3〇°/. Reaction or complete reaction. In several embodiments, at least 2, 5, 丨〇, 2 〇 30 or 50 mg/m 2 of a compound as disclosed herein can be administered = solid. The compound is based on any of the treatment plans disclosed herein. In a particular embodiment, the dosing schedule comprises administering less than 2 mg of turf or a variety of triclinide, trichostatin and related compounds: a drug. In one embodiment, the weekly or sub-administration of itr species or a variety of tricepsin, miscellaneous cililibine and related compounds below the kekeping is repeated. In other embodiments, less than 2 mg/m 2 , 5 mg/m. 10 pg/m 2 and/or 15 mg/m 2 of one or more of trichostatin, triclinylic acid and related compounds ~ 'A '3# 小丨人 j'. In another embodiment, &apos; less than 10 mg/m2 can be administered through a continuous exposure for at least 5 days to the individual. In a specific embodiment, as herein: one or more of trichostatin, triterpene phosphate and related compound bond drugs can be used for cancer, prostate cancer, colorectal cancer and ovarian cancer Treatment. In the present invention, the former (IV) compound and the domain therapeutic agent can be used for preventing and/or treating cancer, sarcoma, lymphoma, white blood or myeloma. In other embodiments of the invention, it is disclosed herein that the Le compound can be used to treat solid tumors. In still other embodiments, the drug-free compound and composition may be used (4) for treatment or cancer, bone, 0, but not limited to cancer of the following organs or tissues, breast, prostate, hard lung, large intestine including But non-restricted large intestine rectum, urinary tract, bladder, non-he 2009 200902034 Jajin's lymphoma, melanoma, kidney, adrenal gland, sputum, pharynx, verrucous gland, stomach, brain and / or ovary. In a particular embodiment, one or more of the precursors of tromethine, triclinide, and related compounds as disclosed herein can be used in the treatment of pancreatic cancer, breast cancer, colorectal cancer, and/or ovarian cancer. In other embodiments of the invention, the compounds disclosed herein are useful in the treatment of angiogenesis-related diseases. In several embodiments, leukemia is treated by continuous infusion of one or more of the precursors of triclinide, triclinide and related compounds prior to continuous infusion for 24, 48, 72 or 96 hours. In other embodiments, continuous infusion can be repeated, for example, every two weeks, every three ten weeks, or at least once every four weeks. In a particular embodiment, a method of treating a tumor, cancer, and other disorder associated with abnormal cell proliferation in a host comprises administering to the host an effective amount of a compound disclosed herein, optionally in combination with a pharmaceutically acceptable form Acceptable carrier. In other embodiments, a prodrug of one or more of triclinide, triclinide, and related compounds as disclosed herein can be used to treat a tumor or cancer that is resistant to one or more drugs, including disclosed herein Examples of tumors or cancers and drugs. In one embodiment, a prodrug of one or more of the sirolimus, tacrolimus and related compounds disclosed herein for use in treating a patient having a drug resistant tumor or cancer, eg, multiple antibodies An effective amount of a drug in a patient with a cancer or cancer, including but not limited to, taxol, rapamycin, tamoxifen, cisplatin, and/or geifeti Gefitinib (iressa) has a drug-resistant tumor or cancer. In one embodiment, one or more of the precursors of tromethine, triclinide, and related compounds disclosed herein, such as 18 200902034, can be administered with an additional chemotherapeutic agent, P- Glycoprotein inhibitors, such as verapamil, cyclosporin (such as cyclosporin A), tamoxetine, calmodulin antagonist, and dessamila Dexverapamil), dexniguldipine, valspodar (PSC 833), biricodar (VX-710), tariquidar (XR9576), left sulqueda (zosuquidar) (LY335979), laniquidar (R101933) and/or ONT-093. In some embodiments, a method is provided comprising administering to a host in need thereof an effective amount of a compound as disclosed herein, or a therapeutically effective amount effective to treat a tumor, cancer, and other disorders associated with abnormal cell proliferation in a host. This includes the pharmaceutical composition of the compound. The invention will be more fully understood by reference to the appended drawings, appended claims, BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the identification of API-2 (Quxiribin) identified as a candidate for Akt inhibitors in the NCI diversity set. Figure 1A shows the chemical structure of API-2 (Quxi Libin). Figure 1B shows the extent of AKT2 phosphorylation in API-2 inhibited by AKT2 transformed NIH3T3 20 cells. The Wile type AKT2-transformed NIH3T3 cells were treated with ΑΡΙ-2 (ΙμΜ) for the indicated time and were subjected to immunoblotting analysis with anti-phosphoric acid-Akt-T308 and -S473 antibodies (top and bottom panels). The figure below shows the performance of the total AKT2. In Figure 1C, three isomorphs of API-2 inhibition of Akt are shown. HEK293 cells were infected with 19 200902034 HA-AkU, HA-AKT2, and HA-AKT3 before EGF stimulation and treated with Αρι 2 (i _ or wortmannin (15 μΜ), cells were lysed and anti-HA antibody Immunoprecipitation. The immunoprecipitate was subjected to in vitro kinase assay (top) and immunoblot analysis with anti-phospho-Akt-T308 (bottom) antibody. Figure 5 shows the performance of Aktl, AKT2, and AKT3 by transfer infection. The id diagram shows that API-2 does not inhibit Akt in the tube. The kinase assay consisting of the active AKT2 recombinant protein in kinase buffer in the assay tube contains API-2 (lane 3). Figure 2 verifies that API-2 will not Inhibition of PI3K, PDK1, and closely related members of the AGC kinase family 10. Figure 2A shows in vitro PI3K kinase assay analysis. In EGF stimulation 4, HEK293 cells were deficient in API-2 (1 μΜ) or Watman The cells were lysed (15 μΜ) for 30 minutes. The cells were lysed and immunoprecipitated with anti-p 11 antibody. The immunoprecipitate was assayed using PI_4_p as the enzyme substrate for in vivo in vivo kinase assay. Figure 2B shows API-2 for in vitro tube 15 PDK1 Effect of activation (above), real The circle shows inhibition by API-2. The open circle shows inhibition by the positive control sturosporine, and Stromostrine is a potent PDK1 inhibitor (IC50 = 5 nM). The following figure shows the immunoblotting analysis of HEK293 cells. HEK293 cells were infected with Myc-PDK1 prior to EGF stimulation and treated with Watmanin or API-2. Immunoblot 20 was detected with the indicated antibody. Figure 2C shows PKCcx with anti-phospho-PKCct-T638 (top panel) And total PKCa (lower panel) antibody followed by immunoblotting analysis of the degree of phosphorylation of API-2 or non-selective PKC inhibitor R〇31 -8220. Figure 2D shows in vitro SGK kinase assay analysis. HEK293 cells EGF was infected with HA-SGK before stimulation and treated with API_2 or Watman 20 200902034. The test tube kinase was immunoprecipitated with HA_SGK using MBP as the enzyme substrate (above). The figure below shows the performance of ha_sgk after metastasis infection. Figure 2E shows the results of the PKA-stimulus assay. The immunopurified PKA is in ADB buffer (Upper Biotech Co., Ltd.) containing the appropriate inhibitor (Αρι_2 or ρκΑΙ) and the enzyme substrate KemPtide ( Ups Cultured in tate Biotechnology Inc)). Quantitative kinase activity. Western blots are shown in Figure 2F. OVCAR3 cells were treated with api-2 for indicated time. The cell-dissolved product was immunized with a suitable anti-phospho antibody (丨 _ 4 map) and an anti-actin antibody (bottom panel). 10 Figure 3 demonstrates that API-2 is inhibited in cancer cells with elevated Akt

Akt activity and cell growth induce apoptosis. Figure 3A shows that the degree of phosphorylation of Western blots 'Akt after treatment with API-2 is detected by anti-phospho-Akt-T308 antibody in a suitable human cancer cell line. The ink spots were again probed with anti-total Akt antibody (bottom panel). In Fig. 3B, the cell proliferation assay is shown. The cell lines indicated in the figure were treated with different doses of API-2 for 24 hours and 48 hours, and then analyzed by the CellTiter 96 Cell Proliferation Assay Kit (Promega). Figure 3C provides an analysis of apoptosis. Cells were treated with API-2, stained with annexin V and PI, and analyzed by FACScan. Figure 4 shows the anti-tumor activity of API-2 in the cancer cell line with elevated Akt, which inhibits Akt downstream targets in mouse xenografts. In Figure 4A, validation of API-2 inhibited Akt phosphorylation of tuberin, Bad, AFX and GSK-30. After treatment with API·2, OVCAR3 cells were lysed and subjected to immunoblot analysis with applicable antibodies. Figure 4B shows 21 200902034 AH-2 inhibits tumor growth. Tumor cells were injected into nude mice with a low left cell concentration and a high right Akt cell concentration. When the tumor reached an average size of about 100-150 cubic millimeters, the animals were treated with vehicle or i mg/kg/曰 API-2. Each measurement represents the average of 1 tumor. Figure 4c 5 shows a representation of mice with 〇vcar3 (right) and OVCAR5 (left) treated with API-2 or vehicle (control). Figure 4D shows an example of tumor size (bottom) and weight (top) at the end of the experiment. In the first plot, the immune dot of the tumor lysate is the anti-phosphorus _Akt_s473 10 in the OVCAR-3-derived tumor treated with APU (T3 and T4) and untreated (T1 and T2). ) and anti-AKT2 (bottom) antibodies were performed. Figure 5 shows that API-2 (Cucidine) inhibits Akt kinase activity in vitro. The in-tube kinase assay was performed using a recombinant strain of PDK1 and Akt in a kinase buffer containing a fat-filled myositol-3,4,5-P3 (PIP3), API-2 and tissue gland H2B as an enzyme substrate. After 30 minutes of incubation, the reaction was separated by SDS-PAGE 15 and exposed to the membrane. Figures 6a-6c provide mRNA and amino acid sequences for human Aktl and also indicate the position of the restriction enzyme. Figures 7a-7d provide mRNA and amino acid sequences of human Akt2, also indicating the position of the restriction enzyme. 20 Figure 8a-8c provides the mRNA and amino acid sequence of human Akt3, also indicating the position of the restriction enzyme. Fig. 9 shows the buffer stability of TCN, TCNP and 5,-0-amidoxime hydrazide tresine citrate (TCNP_Val.). Figure 10 shows the stability of TCN, TCNP and 5,-0- amidoxime-based ursolic acid Quxi 22 200902034 Libin (TCNP_Val.) in liver homogenate. Fig. 11 shows the buffer stability of 5' anthraquinone tromethamine (TCN-Val). Fig. 12 shows the stability of 5'-〇-fibrin fluorenyl triclinide (TCN-Val) in liver homogenate. Figure 13 shows the plasma concentrations of TCN and TCNP after administration of 3 mg TCN (n = 3) in the duodenum. Figure 14 shows the plasma concentrations of TCN and TCNP after administration of 3 mg of τχΝ (η = 3) in the duodenum. 10 Figure 5 shows the plasma concentrations of TCN, TCNP and 6N TCN hexylmercapto derivatives (TCN_1436) after administration of 3 mg of TCN_1436 (n=2) in the duodenum. Figure 16 shows that the plasma concentrations of TCN, TCNP and TCN-Val were 15 degrees after administration of 3 mg of 5,-indole-nonyl fluorenyl triclopibine (TCN-Val) (n=3) via the duodenum. Figure 17 shows the blood coagulation of TCN, TCNP and TCN-Val after administration of 3 mg of 5,0-amidoximephosphoric acid triclinide (TCNP-Val) (n=3) via the duodenum. concentration. [Embodiment; J 20 4. Detailed Description of the Preferred Embodiments In contrast to the prior art and experience, the inventors have determined how to successfully use one or more of tromethine, triterpene phosphate and related compounds before the drug' Treating tumors and cancer in one of the following ways or a combination thereof: (1) one or more of tromethine, trocillin phosphate, and related compounds before 23 200902034 are only administered to a diagnostic test according to the following description. A patient with increased sensitivity; (ii) using a prescribed dosage level that minimizes the toxicity of the drug while still having efficacy; or (iii) using the dosing schedule, which minimizes the toxicity of the drug. 5 The invention encompasses a compound of formula I:

R? and R_2 are each _H, and R2', R3', and R5' are each -H or an amino acid. In some embodiments, R, R2, R2', and R3' are each -H. In other embodiments, R5' is an amino acid. In other embodiments, R2, R2', and R3' are each -H and R5' are amino acids. In another embodiment, the invention encompasses a compound of formula II:

24 15 200902034

Ri and R&gt;2 are each -Η, and each of R2', R3', and R5' is -Η, optionally substituted phosphate or phosphonate (including monophosphate, diphosphate or triphosphate) Or stabilized phosphate precursors). In some embodiments, R!, R2, R2', and R3' are each -Η. In other embodiments, R5' is a phosphate or phosphonate which may be substituted as desired (including monophosphate, diphosphate or triphosphate or a stabilized phosphoric acid precursor). In other embodiments, R!, R2, R2', and R3' are each -H and R5'10 are optionally substituted phosphates or phosphonates (including monophosphate, diphosphate or triphosphate) Or stabilized phosphate precursors). In another embodiment, the invention encompasses a compound of formula III:

15 R3 is a direct bond Ci-C18 alkyl group' and R2', R3', and R5' are each -H. In some embodiments, each of R5', R2', and R3' is -H. In other embodiments, R3 is a linear CrC18 alkyl group. In other embodiments, R5', R2', and R3' are each -H and R3 is a straight 20-bonded Ci_Ci8 alkyl group. In another embodiment, the invention encompasses a pharmaceutical composition comprising a compound of formula I and 25 200902034 pharmaceutically acceptable carrier. In another embodiment, the invention encompasses a pharmaceutical composition comprising a compound of formula II and a pharmaceutically acceptable carrier. In another embodiment, the invention encompasses a pharmaceutical composition comprising Compound 5 of Formula III and a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical composition is suitable for oral administration in another embodiment, and the present invention encompasses a pharmaceutical composition comprising a compound of formula I and a pharmaceutically acceptable carrier, wherein the compound It is present in an amount of from about 1"Nike to about 1000 mg. In another embodiment, the invention encompasses a pharmaceutical composition comprising a compound of formula II and a pharmaceutically acceptable carrier, wherein the compound is present in an amount from about 0.1 nanograms to about 1000 milligrams. In another embodiment, the invention encompasses a pharmaceutical composition comprising Compound 15 of Formula III and a pharmaceutically acceptable carrier, wherein the compound is present in an amount from about 0.1 ng to about 1000 mg. In another embodiment, the invention encompasses a method of treating a tumor or cancer with a mammal comprising administering to a mammal in need thereof an effective amount of a compound of formula I. In another embodiment, the invention encompasses a method of treating a tumor or cancer with a mammal comprising administering to a mammal in need thereof an effective amount of a compound of formula II. In another embodiment, the invention encompasses a method of treating a tumor or cancer with a mammal comprising administering to a mammal in need thereof an effective amount of a compound of formula III 200902034. 5.1 Compounds The present invention provides the use of TCN, TCN-P and related compounds for the treatment of specific treatments for proliferative disorders. In a specific embodiment of the invention, the invention encompasses a composition comprising: (1) a compound of formula IV-VII: f

\

Formula VII 27 10 200902034 wherein R·2', R_3' and R5' are respectively hydrogen, optionally substituted phosphate or root (including an acid-filled, di-phosphoric acid, or tri-acid or stabilized acid-filled prodrug a mercapto group (including a lower carbon group); an alkyl group (including a lower alkyl group); a decylamine, a sulfonate including an alkyl group or an arylalkyl group; a sulfonyl group including a methylsulfonyl group and a benzyl group 5; Wherein, the δ hexyl group may be substituted with one or more substituents as defined in the aryl group described herein; optionally substituted arylsulfonyl; lipids including phospholipids; amines a base acid; a carbohydrate; a peptide; or a cholesterol; or other pharmaceutically acceptable leaving group, which provides a compound in which R2', r3, or heart, respectively, 11 or monophosphate, diphosphorus 10 Acid or triphosphate; wherein R and 1 are respectively hydrogen, optionally substituted phosphate; sulfhydryl (including lower sulfhydryl); decylamine, alkyl (including lower alkyl); aromatic poly An oxygen-expanding group such as polyethylene glycol, an optionally substituted arylsulfonyl group; the lipid includes a wall lipid; Amino acid; carbohydrate; peptide; or cholesterol; 15 or other pharmaceutically acceptable leaving group; in one embodiment, the compound is administered as a 5&apos;-mystery lipid or a 5&apos;-cream lipid. In another embodiment, each of ruthenium and osmium is a linear, branched, or cyclic alkyl group (including a lower alkyl) alkenyl or alkynyl group, a CO-alkyl group, and a CO group, respectively, which may be substituted. -alkenyl, c〇-alkynyl, CO_aryl or heteroaryl, 20 c〇-alkoxyalkyl, CO-aryloxyalkyl, c〇_substituted aryl, sulfonyl, Alkylsulfonyl, arylsulfonyl, or aralkylsulfonyl. In another embodiment 'R2, and R3 are hydrogen. In another embodiment, R2' and R5' are hydrogen. In yet another embodiment, R 2 , , &amp; ' and r 5 are hydrogen. In yet another embodiment, r 2 , r 3 , r 5 , , Ri &amp; r 2 are hydrogen. 28 200902034 in another embodiment Wherein the compound has the structural formula of formula III:

Formula III § κ 10 wherein R3 is hydrazine, optionally substituted straight chain, branched or cyclic alkyl (including lower alkyl), alkenyl, or alkynyl, NH2, NHR4, N(R4)2 An aryl group, an alkoxyalkyl group, an aryloxyalkyl group, or a substituted aryl group; and each R 4 is an anthracene, a fluorenyl group including a lower fluorenyl group, and an alkyl group including a lower alkyl group such as, but not limited to, a fluorenyl group , ethyl, propyl, and cyclopropyl, alkenyl, alkynyl, cycloalkyl, alkoxy, alkoxyalkyl, hydroxyalkyl, or aryl. In one sub-embodiment, R3 is a linear CrC18 alkyl group, an isopropyl group, a tert-butyl group or a phenyl group. In another embodiment, the compounds provided herein have the structural formula of Formula IV and Formula V: 29 200902034

Formula V

In another embodiment, the compounds provided herein have the formula VIII: CH» KK Μ

In another embodiment, the compounds provided herein have the formula IX: 30 200902034

Re I ΚΙ ' M

Wherein 116 is 11, alkyl (including lower alkyl), alkenyl, alkynyl, alkoxyalkyl, hydroxyalkyl, arylalkyl, cycloalkyl, nh2, nhr4, nr4r4, 5 CF3, CH2OH , CH2F, CH2Cb CH2CF3, C(Y3)3, C(Y3)2C(Y3)3, C(=0)0H, C(=0)0R4, C(=0)-alkyl, C(=0) -aryl, C(=0)-alkoxyalkyl, C(=0)NH2, C(=0)NHR4, C(=0)N(R4)2, where each Y3 is H or halo, respectively Atom; and each R4 is fluorene, fluorenyl, including lower fluorenyl, and alkyl includes lower alkyl 10 such as, but not limited to, fluorenyl, ethyl, propyl, and cyclopropyl, alkenyl, alkynyl, cyclic An alkyl group, an alkoxy group, an alkoxy group, a base group, or an aryl group. In one subembodiment, R6 is ethyl, CH2CH2OH or CH2-phenyl. In another embodiment, the compounds provided herein have the formula 15: Formula X: 31 200902034

Wherein R7 is hydrazine, halogen atom, alkyl (including lower alkyl), alkenyl, alkynyl, alkoxy, alkoxyalkyl, hydroxyalkyl, cycloalkyl, nitro, cyanide 5, OH , OR4, NH2, NHR4, NR4R4, SH, SR4, CF3, CH2OH, CH2F, CH2C, 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(di)NH2, C(=0)NHR4, C(= 〇)N(R4)2aN3, where each Y3 is H or a halogen atom; and 10 each R4 is an anthracene, a fluorenyl group includes a lower fluorenyl group, and a sleet group includes a low carbon group such as, but not limited to, a methyl group, a a propyl group, a propyl group, and a cyclopropyl group, an alkenyl group, a fast group, a cyclodextrin group, an alkoxy group, an alkoxy group, a thiol group, or an aryl group. In one sub-example, 'R7 is methyl, ethyl, phenyl, chloro or hydrazine. In another embodiment, the compounds provided herein have the structural formula of formula XI and formula XII: 32 200902034

In another embodiment, the compounds provided herein have the structural formula of formula XIII:

33 200902034 It is to be understood that the compounds disclosed herein may contain the center of the palm. Such a palm center can be either (R) configured or (S) configured or can be a compound thereof. Thus, the compounds provided herein may be enantiomerically pure' or may be a stereoisomeric mixture or a mixture of diastereomers. It is to be understood that the compounds disclosed herein encompass any of the free 5 s form, optically active form, polymorphic form or stereoisomeric form, or mixtures thereof, preferably having the useful properties described herein, and it is well known in the art how to prepare optically active forms, and How to use the standard assays described herein to determine activity, or to use other similar assays known to the art. Examples of methods which can be used to obtain optical isomers of the compounds include the following: 10 (i) Physical separation of crystals A technique for manually separating macroscopic crystals of individual enantiomers. If there are crystals with separate enantiomers, ie the material is a heap and the crystals are visually separated, then this technique can be used; (ii) simultaneous crystallization - only if the material is in a solid state The technique of separating the individual enantiomers from the racemic solution by crystallization; (111) Enzymatic catalyzed optical cleavage - through the different reaction rates of enantiomers and enzymes Techniques for partial or complete separation of racemates; (iv) Enzymatically catalyzed asymmetric synthesis - at least one synthetic step using a 20 enzyme catalyzed reaction to obtain the enantiomerically pure or equivalent of the desired enantiomer Synthetic technology for the synthesis of synthetic precursors; (V) chemical asymmetric synthesis - under the condition that asymmetry (ie, palmarity) can be produced in the product, the desired pair is synthesized by the non-p-precursor precursor a technique for the synthesis of anamorphisms, which can be achieved using a palm catalyst or a palm adjuvant 34 200902034; (vi) separation of diastereomers - racemic compounds with enantiomerically pure reagents (for palms) Adjuvant) reaction 'transfer individual enantiomers to non- Enantiomeric isomers of technology. The diastereoisomers are then converted into more apparent structural differences, so by chromatography or by crystallization separation, the palm adjuvant is subsequently removed to obtain the desired enantiomer; (Vllj趿 趿 趿 笫 及 及 及 及 及 及 及 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称 对称Or the diastereoisomers from the desired enantiomers preferentially crystallize to disturb the equilibrium, and ultimately in principle all materials are expected to be mirrored from the crystallographic diastereomers of the desired enantiomers. The isomer is then released from the diastereomer; / (viii) dynamic optical segmentation ~ this, the enantiomer and the non-external elimination under dynamic conditions, by 15 20 rate, to achieve Part of the optical segmentation compound of the racemate; step light; =) or complete optical segmentation (or (lx) from the non-racemic precursor to the palm starting material to obtain the desired (four) isomer-specific synthesis - by non- At the end of the three-dimensional chemistry, the composition of the composition, the synthesis technology, this (nine) is damaged, Only minimal damage; interaction with the static phase 曰 = remainder 'the enantiomer of the racemate is formed by the palm material by its phase, and the technique of breaking into the liquid phase. Differential interactions; 4 The mobile phase may contain additional pairs of palm materials to stimulate (4) to palm gas chromatography, known as gasification, and enantiomers 35 200902034 by its in the gas phase and containing fixed non- One technique for separating the racemic effect of the column between the palm adsorption phases; (xii) using solvent extraction for palms - separating the enantiomers by dissolving an enantiomer in a specific solvent Structure of the structure; 5 trans-pairing of the palmar membrane - the technique of contact of the racemate with the membrane barrier. The barrier typically separates two miscible fluids, one containing the racemate' driving force such as concentration difference Or a pressure differential causes preferential transport across the membrane barrier. Due to the non-racemic nature of the membrane, only one of the enantiomers in the racemate is allowed to pass through. 10 In several embodiments Offering Quxilide and Phosphoryl Phosphate (TCN-P), 5'-triterpene phosphate (TCN-P), or Trifluendron DMF adduct (TCN-DMF). TCN can be synthesized by any technique known to those skilled in the art, for example In the tetrahedral letter, 49, 4757-4760 (1971). TCN-P can be synthesized by any of the techniques known to those skilled in the art, as illustrated in U.S. Patent No. 4,123,524. Ding 0^-〇] The synthesis of ^? is described, for example, in INSERM, 81, pages 37-82 (1978). Other TCN-related compounds as described herein can be synthesized, for example, according to the methods disclosed below: Gudmundsson KS et al., "2', 3 '-Di-deoxysangivamycin, 2',3'-dideoxy 1 〇丫〇〇 &amp; 11^(^11' and 2',3'-di-deoxytricidin, nucleoside, nucleus 20 Glycosidic acid, nucleic acid '20(10-11): 1823-1830 (October-November 2001); Porcari AR et al., "6-N-fluorenyl triclinbine analogue: thiol carbon chain length and resistance Relationship between structure and activity of HIV-1 activity", J Med Chem, 43(12): 2457-2463 (June 15, 2000); Porcari AR et al. 'Acyclic agarose analogue of Quxilide: Lack of antiviral and anti-proliferative Sex and low intracellular phosphorus 36 200902034 Acidification has an interaction relationship "'nucleosides, nucleotides, 18 (11-12): 2475-2497 (November-December 1999); Porcari AR· et al., "Qu Xi Li Phthalas deoxysaccharide analogues: interaction between antiviral and antiproliferative activities and low intracellular squaring. J Med Chem, 43(12): 2438-2448 (June 15, 2000); 5 P0rcari A. R_ et al. 'Synthesis and antiviral activity of 2-substituted analogues of trifluralin”, nucleosides, nucleotides, nucleic acids, 22(12): 2171-2193 (December 2003); Porcari A_R· et al., "Improved total synthesis of Quxi Libin: Tricyclic nucleosides with anti-proliferative and antiviral properties", nucleosides, nucleotides, nucleic acids, 23(1-2): 31-39 (2004); Schweinsberg PD et al., "Identification of Metabolites of Antiviral Tris 10 Cyclic Nucleosides (NSC-154020)", Biochem Pharmacol, 30(18): 2521-2526 (September 15, 1981); Smith KL et al., "Synthesis of novel 2'-β- (:- mercapto-related synthesis of trifluralin analogs) as an anti-HCV agent, Bioorg Med Chem Lett, 14(13): 3517-3520 (2004 7 Month 5 曰); Townsend LB et al "Synthesis and biological activity of certain nuclear rings corresponding to five-ring, six-ring and its I5", Nucleic Acids Symp. Ser., 1986(17): 41-44 (1986); and/or Wotring L丄· et al., “Tursilyl phosphate (TCN-P) acts as an activation machine for the pre-TCN drug-dissipation form”, Cancer Treatment Rep. '70(4): 491-7 (June 1986). In another preferred embodiment, the compounds provided herein have the structural formula of formula XIV as follows: 37 200902034

Hj

In another preferred embodiment, the compounds provided herein have the structural formula of Formula XV: CM3

Formula XV In another preferred embodiment, the compounds provided herein have the formula of formula XVI: 38 200902034

In the case where the compound is sufficiently basic or acidic to form a stable non-toxic acid salt or an alkali salt, the compound is administered as a pharmaceutically acceptable salt. Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic acids or acids. Suitable salts include salts derived from metals such as I and Na, soils such as earth and magnesium, and various other acids are well known in the art. In particular, examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids which form physiologically acceptable anions, such as 10 sulfonate, methanesulfonate, acetate, citrate , malonate, tartrate, succinate, benzoate, ascorbate, alpha-ketoglutarate, and alpha-glycerophosphate. Suitable inorganic acid salts can also be formed, including sulfates, citrates, bicarbonates, and double salts. Pharmaceutically acceptable salts can also be obtained using standard procedures well known in the art, for example, by fully testing a compound such as an amine with an appropriate acid which forms a physiologically acceptable anion. Alkali metal (e.g., sodium, potassium or lithium) or phytochemical (e.g., mom) salts of carboxylic acids can also be made. Any of the nucleonates described herein may be administered as a precursor of a sulphate acid. 20090203434 High activity, bioavailability, stability, or otherwise altering the nature of the oath. A variety of nucleotide precursor ligands are known. Substantially nucleus: alkylation, deuteration or other lipophilic modification of one of scaly acid, diphosphoric acid or triphosphate will increase the stability of the nucleotide. Examples of substitutions of one or more hydrogens on the replaceable phosphoric acid moiety are based on the group, aryl, steroid, carbohydrate, including sugars, 1,2-bristyl glycerol, and alcohols. A variety of substituents are described in R. j〇nes and N.

Bischofberger, Antiviral Research, 27 (1995) 1-17. Either of these can be used in combination with the disclosed nuclear oath to achieve the desired effect. In one embodiment, triclinide or a related compound is provided as a 5,-10 hydroxy lipophilic precursor. A non-limiting example of a U.S. patent that discloses covalently incorporated in a nucleoside, preferably in a nucleoside or lipophilic formulation, is disclosed in U.S. Patent No. 5,149,794. (September 22, 1992, Yatvin et al.), 5,194,654 (March 16, 1993, Hostetler et al.), 5,223,263 (June 29, 1993, Hostetler et al. 15), 5,256,641 (October 26, 1993) , Yatvin et al., 5,411,947 (May 2, 1995 Host 'Hostetler et al.), 5,463,092 (October 31, 1995, Hostetler et al.), 5,543,389 (August 6, 1996, Yatvin et al.), 5,543,390 ( On August 6, 1996, Yatvin et al., 5,543,391 (Aug. 6, 1996, Yatvin et al.), and 5,554,728 (September 10, 1996, 20 Basava et al.) were incorporated by reference. At the office. The disclosure of a lipophilic substituent or lipophilic preparation that can be attached to the tromethine or related compound of the present invention includes W〇89/02733, W0 90/00555, WO 91/16920, W0 91/18914 , WO 93/00910, W0 94/26273, WO/15132, EP 0 350 287, EP 93917054.4, 40 200902034 and WO 91/19721. Additional non-limiting examples of derivatives of triclinide or related compounds are such compounds containing the substituents described in the following publications. Such derived trichostatin or related compounds can be used for the indications described herein, and 5 for use as an antiviral agent, including as an anti-HIV agent or an anti-HBV agent. Ho DHW (1973) Distribution of kinases and deaminase in 人β-D-arabinofuranosylcytosine in human and mouse tissues' Cancer Research, 33, 2816_282〇; Holy A. (1993) Phosphorus Modified Nucleotide Analogs, De Ciercq (eds.) 'Advances in Antiviral Drug Design', Issue 1, jAi Press, 179-231 10 pages; Hong CI, Nechaev, A., and West, CR (1979a Synthesis of a 1β-3-arabinofuranosylcytosine conjugate of cortisol and corticosterone and its tumor activity, Biochem Biophys Rs Commun, 88: p.1223-1229; Hong CI·, Nechaev, A· , Kirisits, AJ Buchheit, DJ and West, CR (1980) nucleoside conjugates as potential anti-tumor agents; 3. 15 corticosteroids and selected lipophilic alcohols - φ-D-arabinofuranosyl) Synthesis and antitumor activity of pyrimidine conjugates, J. Med. Chem. 28, 171-177; Hostetler Κ_Υ·, Stuhmiller, L_M., Lenting, Η·Β·Μ. van den Bosch, H. and Richman, DD (1990) Synthesis and anti-retroviral activity of azidothymidine and other antiviral nucleoside phospholipid analogs, J. 20 Biol. Chem. 266, 1171 4-11717; Hostetler KY, Korba, B., Sridhar, C., Gardener, M. (1994a) Antiviral activity of fat-filled-dideoxycytidine in Hepatitis B-infected cells and increased liver in mice Ingestion, Antiviral Res, 24, 59-67; Hostetler KY, Richman, DD, Sridhar, CN Feigner, PL, Feigner, J., Ricci, J., Gerdener, 41 200902034 MF, Selleseth, DW and Ellis, MN (1994b Phospholipid azido thymidine and phospholipid-ddC: uptake in mouse lymphoid tissues and evaluation of antiviral activity in human immunodeficiency virus-infected cells and mice infected with Lloyd's disease virus, antimicrobial chemotherapy , 38, 2792-2797; Hunston 5 RN, Jones, AA, McGuigan, C., Walker, RT, Balzarini, J., and De Clercq, E. (1984) Derived from 2'-deoxy-5-fluoro Synthesis and biological properties of several cyclic phosphotriesters of uridine, J Med Chem, 27, 440-444; Ji Υ·Η., Moog, C., Schmitt, G., Bischoff, Ρ··Luu, B. (1990); 7β-hydroxycholesterol and a pyrimidine nucleoside phosphodiester as 10 potential antitumor agents; synthesis and anti-tumor Preliminary assessment of activity, J Med Chem, 33, 2264-2270; Jones AS, McGuigan, C., Walter, RT, Balzarini, J. and DeClercq, E. (1984) Several nucleoside cyclic squamous amino acids Synthesis, properties and biological activity, J. Chem. Soc. Perkin Trans. I, 1471-1474; Juodka BA and Smart J., (1974) ditribonucleoside a (PCN) amino acid derivative Synthesis, Coll. Czech. Chem. Comm. 39 » 363-968 ; Kataoka S. ' Imai, J. &gt; Yamaji, N., Kato, M., Saito, M., Kawada, T. and Imai, S (1989) Alkylation of cAMP derivatives, selective synthesis and biological activity, nucleic acid research Sym. Ser., 21, 1_2; Kataoka S., Uchida, R. and Yamai, Ν· (1991) 2 〇 adenosine 3' Convenient synthesis of 5' cyclic phosphate (cAMP) benzyl triester and methyl triester, heterocycle 32, 1351-1356; Kinchington D., Harvey, JJ, O'Connor, TJ, Jones, BCNM, Devine, KG, Taylor-Robinson, D_, Jeffries, D.J_ and McGuigan, C_ (1992) zidovudine filled with tyrosine and chlorin derivatives in test tubes 42 200902034 Anti-HIV and ULV Disease resistance Comparison of the efficacy of antiviral chem.

Chemother. 3 '107-112, Kodama Κ·, Morozumi, Μ., Saitoh, Κ.Ι., Kuninaka, Η., Yoshino, Η· and Saneyoshi, Μ· (1989) 1 -β-D-arabinose furan Antitumor 5 activity and pharmacology of glycosylcytosine-5,-stearyl phosphate: an orally active derivative of Ι-β-D-arabinofuranosylcytosine, Journal of Cancer Research in Japan, 80, 679-685 Korty M. and Engels J., Adenosine- and guanosine 3', 5'-acid benzyl esters on ventricular myocardium, Naunyn-Schmiedeberg, s Arch Pharmacol 310, 103-111; Kumar A., Goe, PL, J〇nes, aS, Walker, RT, 10 Balzarini, J. and De Clercq, E., (1990) Synthesis and biological evaluation of several cyclic phosphonium nucleoside derivatives, J. Med. Chem. 33, 2368-2375; LeBec C. and Huynh-dinh T. (1991), synthesis of 5-fluorouridine and arabinose-reactive lipophosphate triester derivatives as anticancer prodrugs, tetrahedron Letters 32, 6553-6556; Lichtenstein J., Barner, HD and Cohen 15 SS (1960), exogenously supplied nucleotides by E. coli, J. Biol. Chem. 235, 457-465; Lucthy J., Von Daeniken, A., Friedwich, J. 'Manthey, B., Zweifel, J., Schlatter, C. and Benn, MH (1981), Synthesis of three natural cyanothiosulfans And toxicological properties, Mitt Geg Lebensmittelunters Hyg, 72, 131-133 (Chem. Abstr. 95, 20 127093); McGuigan C., Tollerfield, SM and Riley, PA (1989), several phosphotriesters of the antiviral drug Ara Synthesis and Bioassay of Derivatives, Nucleic Acid Research 17,6065-6075; McGuigan C., Devine, KG, O'Connor, TJ, Galpin, SA, Jeffries, DJ and Kinchington, D. (1990a), as an Anti-HIV Compound 3'-azido-3'-43 200902034 Synthesis and evaluation of several novel phosphoproline derivatives of deoxythymosin (AZT), antiviral chemotherapy 1,107-113; McGuigan C., O 'Connor, TJ, Nicholls, SR, Nickson, C. and Kinchington, D. (1990b), Synthesis of several substituted dialkyl phosphate derivatives of AZT and ddCyd and 5 anti-HIV activities, antiviral chemotherapy 1, 355-360 ; McGuigan C·,

Nicholls, SR, O'Connor, TJ· and Kinchington, D. (1990c) Synthesis of several novel dialkyl phosphate derivatives of 3'-modified nucleosides as potential anti-AIDS drugs, antiviral chemotherapy 1 , 25-33; McGuigan C., Devine, KG, O'Connor, TJ· and Kinchington, D. (1991), several of l〇3'-azido-3'-deoxythymidine (AZT) Synthesis and Anti-HIV Activity of Haloalkylphosphoric Acid Derivatives: Strong Activity of Tri-Ethyl Ethoxypropylamine Mercapto Compound, Antiviral Research 15,255-263; McGuigan C., Pathirana, RN, Mahmood, N., Devine, KG and Hay, AJ (1992) 'AZT's arylphosphoric acid derivatives retain anti-HIV1 activity in cells resistant to AZT, Antiviral Research 17,311-321; McGuigan C., Pathirana, R_N·, Choi, SM'Kinchington, D. and O'Connor, TJ (1993a) 'AZT phosphoproline derivatives as HIV inhibitors: research on reciprocal end groups, antiviral chemotherapy 4,7 -101 ; McGuigan C.,

Pathirana, RN, Balzarini, J. and De Clercq, E_ (1993b), intracellular delivery of a biologically active AZT nucleotide by AZT aryl acid derivative, j. Me (j. Chem. 36, 1048- 1052. The alkyl-hydrogen phosphonic acid derivative of the anti-HIV agent AZT is less toxic than the parent nucleoside analog. Antiviral chemotherapy 5,271-277; Meyer RB, Jr·, Shuman, DA and Robins, RK ( 1973), ° Tickets Reconciliation 3, 5, - Ring 44 200902034 Synthesis of squarenine, tetrahedral letter, 269-272; Nagyvary J., Gohil, R_N., Kirchner, CR and Stevens, JD (1973) Studies on cyclic AMP neutral esters, Biochem Biophys Res Commun. 55, 1072-1077; Namane A., Goyette, C., Fillion, MP, Filion, 5 G and Huynh-Dinh, T. (1992) 'AZT modified brain delivery using a glycosyl-triester precursor, J. Med. Chem. 35'3939-3044; Nargeot J., Nerbonne, JM, Engels, J. and Leser, Η·Α· ( 1983), Natl. Acad. Sci. USA 80, 2395-2399; Nelson, Κ·Α., Bentrude, WG, Stser, WN and Hutchinson, JP (1987), nucleoside ring 3, 5,-10 — Phosphoric acid ring The problem of the balance of the chair-like distortion, 1H NMR and X-ray crystallography of the diastereoisomers of the thymidine 3',5'-monophosphate, J. Am. Chem. Soc. 109,4058 -4064 ; Nerbonne J.M_, Richard, S., Nargeot, J. and Lester, HA (1984), novel photoactivated cyclic nucleate producing intracellular intracellular concentrations of cyclic A Μ P and cyclic G Μ P Jump, 15 Nature 301, 74-76; Neumann J_M., HervS, M., Debouzy, JC, Guerra, FI, Gouyette, C., Dupraz, Β· and Huynh-Dinh, T. (1989), Thymidine Synthesis of Glucosyl Phospholipids and Transmembrane Transport by NMR, J. Am_ Chem. Soc. 1U, 4270-4277; Ohno R., Tatsumi, N., Hirano, M., Imai, K., Mizoguchi, H. , Nakamura, 2〇T., Kosaka, M., Takatuski, K, Yamaya, T., Toyama, K., Yoshida, T., Masaoka, T., Hashimoto, S., Ohshima, T., Kimura, I , Yamada, K. and Kimura, J. (1991) 'Treatment of spinal myelodysplasia with oral Ι-β-D-arabinofuranosylcytosine-5'-stearyl sulphate, Oncology 48,451 -455 ; Palomino E., Kessle, D. 45 200902034 And Horwitz, JP (1989), 2', 3'-dideoxynucleosides delivered to the brain in a dihydrogen 0 to ° carrier system, J. Med. Chem. 32, 622-625; Perkins RM , Barney, S., Wittrock, R., Clark, PH, Levin, R., Lambert, DM, Petteway, SR, Serafinowska, HT, Bailey, 5 SM, Jackson, S., Harnden, MR, Ashton, R· , Sutton, D., Harvey, JJ and Brown, AG (1993), in mice BRL47923 and its oral prodrug SB203657A against Lloyd's murine leukemia virus infection, antiviral study 20 (Suppl.I) 84 ; Piantadosi C ., Marasco, CJ, Jr., Morris-Natschke, SL, Meyer, KL, Gumus, F., Surles, 10 JR·, Ishaq, KS, Kucera, LS, Iyer, N., Wallen, CA, Piantadosi, S And Modest, EJ (1991) Synthesis and Evaluation of Novel Ether Lipid Nucleoside Defects Against HIV-1 Activity, J. Med. Chem. 34, 1408-1414; Pompon A., Lefebvre, I., Imbach, JL , Kahn, S. and Farquhar, D_ (1994), azido thymidine-5'-monophosphate--and 贰15 (p-pentyloxymethyl) ester in cell extracts and tissues Decomposition path of nutrient base: application of online ISRP-clean HPLC technology, antiviral chemotherapy 5 ' 91-98 ; Postemark, T · (1974) cyclic AMP and cyclic GMP, Anu. Rev. Pharmacol. 14,23 -33 ; Prisbe, EJ., Martin, JCM, McGee, DPC·, Barker, MF, Smee, DF, Duke, AE, 20 Matthews, TR and Verheyden, JPJ (1986), 9-[(l, 3- Synthesis and Anti-Tudouvirus Activity of Phosphate and Phosphonic Acid Derivatives of Phenyl-2-propoxy)methyl]guanine, J. Med. Chem. 29,671-675; Pucch F., Gosselin, G. , Lefebvre, I., Pompon, Α·, Aubertin, AM, Dirn, A. and

Imbach, JL (1993), nucleoside monophosphate via reductase vector activation pathway 46 200902034 Intracellular delivery 'antiviral research 22 ' 155-174; Pugaeva VP, Kochkeva, S.I_, Mashbits, FD and Eizengart, R.S_ (1969), Toxicological Evaluation and Health Standard Rating for Ethyl Sulfide in Industrial Atmosphere, 〇1§·Trf. Prof. Zabol. 13 » 47-48 (Chem.Abstr.72, 212) ; Robins 5 RK (1984), the potential of nucleotide analogues as a retrovirus and tumor suppressor, Pharm. Res. 11-18; Rosowsky A., Kim, SH, Ross and J. Wick, MM (1982), pro 5'-(alkylphosphoryl) ester of lipid Ι-β-D-arabinofuranosylcytosine and its N4-mercapto and 2,2'-anhydro-3'0-fluorenyl derivatives as possible precursors Drugs, J. Med. Chem. 25, 171-178; Ross 10 W. (1961), Walker equipment for increased sensitivity to pre-glucose pre-treatment with aromatic branched aromatic nitrogen fenium gas, Biochem. Pharm. 8, 235-240; Ryu Ε·Κ·, Ross, RJ, Matsushita, Τ·, MacCoss, Μ·, Hong, CI and West, C.R_ (1982), phospholipid-nucleoside conjugates 3, Ι-β-D-A Synthesis and preliminary bioassay of primosefuranosylcytosine 5' diphosphate [-],2-dimercaptoglycerol 15 , J. Med. Chem. 25, 1322-1329 ; Saffhill R· and Hume WJ (1986) , 5-iodooxyuridine and 5-bromodeoxyuridine are separated by serum from different sources and the results of their use in binding to DNA, Chem. Biol. Interact. 57, 347-355; Saneyoshi, Μ., Morozumi, Μ., Kodama, K., Machida, J., 20 Kuninaka, A. and Y〇shino, H. (1980) Synthetic nucleosides and nucleosides XVI — Series Ι-β-D - Synthesis and biological evaluation of arabinofuranosylcytosine 5,-alkyl or arylphosphoric acid, Chem. Pharm. Bull. 28, 2915-2923; Sastry, JK, Nehete, Ρ·Ν·, Khan, S. , Nowak, BJ, Plunkett, W., Arlinghaus, RB and Farquhar, D. (1992) 'Transpermeable membranes of deoxygenated 47 200902034 uridine 5'-monophosphate analogues inhibit human immunodeficiency virus infection, Mol Pharmacol. 41,441-445; Shaw JP, J〇nes, RJ, Arimilli, MN, Louie, MS, Lee, WA· and Cundy, KC· (1994), PMEA from PMEA Precursor Oral Bioprofit 5 Usage Rate of the Bora Lali Rat, AAPS 9th Annual Conference, San Diego, CA (Abstract); Shuto S., Ueda, S., Imamura, S., Fukukuawa, K., Matsuda, Α· and Ueda, T. (1987), a convenient one-step synthesis of 5'-phospholipid nucleosides by enzyme-catalyzed two-phase reaction, tetrahedral letter 28, 199-202; Shuto S. 'Itoh, H., Ueda, S., Imamura, S., Kukukawa, K., Tsujino, Μ·, Matsuda, 10 A_ and Ueda, T. (1988), Convenient enzymes for 5'-(3-sn_phospholipid) nucleosides Catalytic Synthesis and Its Anti-Leukemia Activity, Chem. Pharm. Bull. 36, 209-217. A preferred phosphoric acid precursor group is S-mercapto-2-thioethyl, also known as "SATE". Additional examples of prior use of the disclosed drugs are described in the following patents and patent applications: U.S. Patent Nos. 5,614,548, 5,512,671, 5,770,584, 5,962,437, 5,223,263, 5,817,638, 6,252,060, 6,448,392, 5,411,947, 5,744,592, 5,484,809, 5,827,831, 5,696,277, 6,022,029 ' 5,780,617 &gt; 5,194,654 ' 5,463,092 &gt; 5,744,461 ' 4,444,766, 4,562,179,4,599,205, 4,493,832, 4,221,732, 20 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 &lt;6,670,341 &gt; 2004/0161398, 2002/082242, 5,512,671, 2002/0082242 and/or PCT Publication WO 90/11079, WO 96/39197, and/or WO 93/08807. 48 200902034 5.3 Definitions As used herein, the terms "cancer" and/or "cancerous" describe or describe a physiological condition characterized by unregulated cell growth in a mammal, that is, a proliferative disorder. Examples of such proliferative disorders include cancers such as carcinomas, lymphomas, blastomas, sarcomas, and leukemias, as well as other cancers disclosed herein. More specific examples of such cancers include breast cancer, prostate cancer, colorectal cancer, squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, gastrointestinal cancer, sputum cancer, cervical cancer, Indian cancer, liver cancer such as liver cancer , bladder cancer, colorectal cancer, endometrial cancer, kidney cancer and squamous cell carcinoma. 10 Non-limiting examples of other cancers are basal cell carcinoma; cholangiocarcinoma; bone cancer, brain and central nervous system (CNS) cancer; choroidal carcinoma; connective tissue cancer; esophageal cancer; eye cancer; head and neck cancer; gastric cancer; Tumor; pharyngeal carcinoma; lymphoma including Hodgkin's lymphoma and non-Hodgkin's lymphoma; melanoma; myeloma; neuroblastoma; oral cancer (eg lip, tongue, mouth and throat); Carcinoma; retinoblastoma; rhabdomyosarcoma; rectal cancer; respiratory cancer; sarcoma; skin cancer; gastric cancer; testicular cancer; uterine cancer; urinary system cancer and other cancers and sarcomas. As used herein, the term "tumor" refers to all neoplastic cell growth and proliferation, including malignant or benign and all precancerous and cancerous cells and tissues. Example 20 For example, a particular cancer can be characterized by a solid mass of tumor. A solid tumor mass can be a primary tumor mass when present. Primary tumor mass refers to the growth of cancer cells within the tissue resulting from normal cell transformation of the tissue. In most cases, the primary tumor mass is identified by the presence of a cyst, which can be found by visual methods or palpation methods, or by the irregular shape, texture or weight of tissue 49 200902034. Cyst. However, several primary tumors cannot be palpated and can only be detected by medical imaging techniques such as X-rays (such as mammography) or by needle aspiration. The use of the techniques described later is more common in early detection. Molecular analysis and phenotypic analysis of cancer cells within the tissue usually 5 can verify whether the cancer is a cancer that is endogenous to the tissue, or whether the lesion is from another part of the cancer metastasis. As used herein, unless otherwise indicated, the term "alkyl" includes, for example, c, to Cm straight, branched or cyclic first hydrocarbons, second hydrocarbons, or third smoke, particularly including methyl, trifluoromethyl. , ethyl, propyl, isopropyl, cyclopropyl, 10 butyl, isobutyl, tert-butyl, pentyl, cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl , cyclohexylmethyl, 3-methylpentyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl. The alkyl group may, for example, be substituted with one or more substituents such as a halogen atom (F, α, Br or I), for example, CF3, 2-Br-ethyl, CH2F, CH2C1, CH2CF3, or 15 CF2CF3), hydroxyl (eg CH2〇H), amine (eg CH2NH2, CH2NHCH3, or CH2N(CH3)2), alkylamino, arylamine, alkoxy, oxy, nitro, gas Base (e.g., CH2N3), cyano (e.g., CH2CN), sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate, either unprotected, or as known to those skilled in the art, may be protected as desired, e.g., Greene Etc. 2 有机 有机 organic synthesis 蒦 蒦 ,, 1991, 2nd edition, about the round of Willie & Sons, New York's teachings 'by reference here. As used herein, the term lower alkyl, as used herein, refers to a saturated linear, branched, or suitably cyclic (eg, cyclopropyl)alkyl group, including substituted forms, and Unsubstituted form. 50 200902034 The term "alkylamino" or "amino" includes amino groups which have one or two alkyl or aryl substituents, respectively. The term amino acid includes both natural and synthetic (X, β, γ or δ amino acids, including but not limited to amino acids present in proteins, ie glycine, alanine, • 5 valine, white Aminic acid, isoleucine, methionine, phenylalanine, tryptophan, valine, serine, threonine, cysteine, tyrosine, aspartame, glutamine, Aspartic acid, glutamic acid, lysine, arginine, and histamine (acid. In the preferred embodiment, the amino acid is in the L-configuration. In addition, the amino acid can be an alanine, anthracene. Aminyl, leucine, isoleamine, amidoxime, Q, phenylpropylamine, tryptamine, egg oxime, glycidinyl, amidoxime, Susamine, cysteamine, tyramine, aspartame, glutamine, aspartame, pentane, amidoxime, spermine sulfhydryl, histamine sulfhydryl , amphetamine, ρ-amidoxime, β-leucine thiol, β-iso-amine amine, Mt surface group, β-phenyl (tetra) fluorenyl, 卩 _ color _ base, Ρ _ egg Amine • 15 rpm, β·glycidyl sulfhydryl, β-silylamine, β-threonine, β-cysteamine醯 (yl, tyramine sulfhydryl, Ρ-aspartate, β- face amine, β-aspartate, Ρ-pentamethylene, β·aminoxime, β_spermine Derivation of a group or a group of amine sulfhydryl groups. When using amino acid-words, they are specifically disclosed by (IV) L-grade or synthetic amino acid, including but not limited to D configuration and [configuration ^, β , γ or δ glycine, alanine 'imidic acid, leucine, isoleucine, methionine, phenylalanine, tryptophan, valine, serine, threonine, half Cystamine, citrate, aspartate, faceamine, aspartic acid, tyrosine, lysine, arginine and histidine. Unless otherwise defined, "protected" as used herein词包51 200902034 立二κ' nitrogen, sulfur or atom-filling to prevent its further reaction or for the purpose of the group. A wide variety of oxygen and nitrogen protecting groups are known for their organic synthesis skills | Reference G job eAWuts, Organic Synthesis Base '1999, 3rd edition, John Wiley & Sons, New York). ', 5, unless otherwise stated, aryl-words as used herein include stupid, biphenyl Or a naphthyl group and preferably a phenyl group. The aryl group may be optionally substituted with one or more moieties such as a halogen atom, a trans group, an amine group, an alkylamino group, an arylamine group, and a pyrene group. Oxyl, aryloxy, sulphate, cyano, sulphate, sulphate, phosphonic acid, citrate or phosphonate, either unprotected or as known to those skilled in the art, may be protected as desired, for example, as Greene Et al., Organic Synthetic Protection Group, 1991, 2nd ed., John Wiley & Sons, New York teaches that 0 or a aryl group includes a aryl group-based aryl or aryl group. The term "alkyl" includes aryl having an alkyl substituent. The term halo is used herein to include chloro, bromo, iodo, and fluoro. The term thiol includes a carboxylic acid ester wherein the non-carbonyl moiety of the ester group is It is selected from a linear chain, a branched chain or a cyclic alkyl group or a lower alkyl group, an alkoxyalkyl group including a methyl group, a group including a sulfhydryl group, an aryloxy group such as a phenoxy fluorenyl group, The aryl group includes a phenyl group, a sulfonic acid ester such as an alkyl group which may be substituted with a halogen, (^ to (: 4 alkyl or 〇^ to (4 alkane 20 oxy group) as needed. A sulfonyl or aralkyl sulfonyl group includes a fluorenyl group, a mono- sulphonate, a di-salt ester, or a tri-salt ester, a triphenyl fluorenyl group or a monomethoxytriphenyl fluorenyl group, a substituted section a base, a tricalcium base (for example, a dinonyl-t-butyldecylalkyl group) or a diphenylmethyldecyl group. The aryl group of the ester preferably contains a phenyl group. The term "low carbon thiol" refers to a fluorenyl group in which the non-carbonyl moiety 52 200902034 is a lower alkyl group. As used herein, with respect to enantiomeric purity, the term "substantially free" or "substantially absent" means that a composition comprises at least 85% or 90% by weight, preferably 95% to 98. % by weight, more preferably 99% to 100% by weight of the specified enantiomer. In a preferred embodiment, in the methods and compounds of the invention, the compound is substantially free of other enantiomers. Similarly, the term "separation" means a compound composition comprising at least 85% or 90% by weight, preferably 95% to 98% by weight, and still more preferably 99% to 100% by weight of the compound, the difference comprising Other chemical species or enantiomers. The term "respectively separate" is used herein to mean a variable that is applied separately, and the variable varies independently depending on the application. Thus, in a compound such as R''XYR", where R" is "carbon or nitrogen, respectively", two R" may be carbon, two R" may be nitrogen, or one R" is carbon and the other R" is nitrogen. 15 The term "pharmaceutically acceptable salt or prodrug" is used in the preceding description to describe a pharmaceutically acceptable form of a compound (such as an ester, phosphate, ester or related group) which is administered to a patient. This compound is available upon request. Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic bases and inorganic or organic acids. Suitable salts include salts derived from alkali metals 20 such as clocks and sodium, and salts derived from soils such as magnesium and magnesium, and a variety of other acids are well known to those skilled in the pharmaceutical industry. A pharmaceutically acceptable prodrug means a compound which is metabolized in a host, such as by hydrolysis or oxidation, to form a compound of the invention. Typical examples of prodrugs include compounds having a biolabile protecting group on the functional portion of the active compound. Prodrugs include 53 200902034 which can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, hydrolyzed, alkylated, dealkylated, deuterated, deuterated, phosphorylated to produce active compounds. Compound. The term "pharmaceutically acceptable esters" as used herein, unless otherwise stated, includes esters of one or more compounds which are suitable for use in the context of prudent medical judgment. Contact with the host tissue without any major, sexual, irritating, allergic reactions, etc., with reasonable benefits / risks 勹 effective for its intended use. The term "individual" as used herein refers to an animal, preferably mammal 10 is optimally human. Mammals include non-human mammals including, but not limited to, pigs, sheep, goats, cows, deer, baboons, horses, monkeys, and other non-human primates, dogs, cats, rats, mice, rabbits, or any other known Or a mammal as disclosed herein. As used herein, the terms "tremidine prodrug, TCN, TCN-oxime, and related compound 15" refer to such compounds disclosed herein, such as the formula ι_χνΐ compounds, and salts thereof, and hydrates thereof. In some embodiments, the words "tresiformin prodrug" or "cursillabine prodrug" are used interchangeably to mean, for example, the compounds of formulas I, II, III, XIV, XV and XVI and their salts and their hydration. Things. 6. In Vivo Efficacy/Dosing Acupuncture 20 In another aspect of the present invention, a dosing schedule is provided which limits the toxic side effects of the triclinide prodrug, TCN, TCN-P and related compounds. In one embodiment, such a dosing schedule reduces or eliminates toxic side effects including but not limited to hepatotoxicity, thrombocytopenia, hyperglycemia, vomiting, hypoxemia, anemia, hypoalbuminemia, myelosuppression, blood Triglyceride 54 200902034 The ester is too high, the starch in the blood is too high, the spleen, the yin, the yin and/or the hair; In another embodiment, the administration of crystallization 70 cilostane precursor, TCN, TCN-P, and related compounds is provided to at least a partial or complete reaction of at least 15% _2% of the individual in a sturdy manner. For specific 胄 /. Individual to ^ . ις 0Λ .. , in the example, part of the reaction may be at least 15, 2G, 25, 3G, 35, 4 ^ 70, 75, 80 or 85% of the tumor. In Fu Xi, 55, 60, 65, and a consistent example, at least 15 10 15 20 of the treatments were treated with a response of 3 〇 '35, 40, 50, ^ _ or 9 〇 ° / °. In other embodiments, any of the treatment vesicles disclosed herein can be used. W丨 obtains this reaction ratio. .匕 歹 , 1, 日 已经 已经 已经 , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , 个体 , , , 个体 , The dosing schedule includes administering the drug once a week to a week;;, ra, week, followed by a week of undosing (ie, through a 28-day cycle). . In the case of sputum, this 28-day cycle can be repeated at least 2, 3, 4, or 5 MU, or repeated until the tumor is regressed significantly. In other embodiments, a 42-day cycle is provided wherein the compound disclosed herein can be weekly-timed for four weeks, followed by two weeks of unmedicated time. In other embodiments, such a 42 day cycle can be repeated at least 2, 3, 4, or 5 times or repeated until the tumor is significantly degraded. In a particular embodiment, less than 12, less than 11 or less than 10 mg/m2 of trichostatin prodrug, TCN, TCN-P and related compounds can be administered according to a 42 day period. In other specific embodiments, '2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 mg/m 2 of triclinide prodrug, TCN, TCN-P, and related compounds, 15 , 20, 25 55 ' 60 ' 65, 70, 75, 80 55 200902034 According to the 42-day cycle. In another embodiment, the method of treating the cancer by the individual is based on the application of the sedative-type music program 'the mother's weekly dose of 10 mg/m2 or less of Qucilide to 5 10 15 20 Zha Yuxi - every &gt; then drugs, tcn, tcn_p and related combinations

Things. In particular, I and H are difficult to give 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5 5, 8^^6, 6·5, 7 , 7.5 , 8 , 8.5 , 9 , 9.5 or 10 mg / m 2 as herein TCN-P and related compounds. ^ Qucilide prodrug, double, in the embodiment of the present invention, TPM, disclosed herein The triclinide prodrug, the target compound can be administered in a single-dose dose at a short time, for example, about 5, 1 〇, 15, medium: minute time. In other embodiments, a feeding plan is provided, and the towel is provided. The compound is administered by continuous rounds of injection to &gt;, m96 or call, and time (4). In some embodiments, it can be repeated at a certain frequency by continuous injection or high dose (four) administration. The frequency of administration is at least: per Week-time, every two weeks-times, every three weeks, once a month, once every five weeks, every six weeks, every eight weeks - two every ten weeks ~ owed, and / or every twelve weeks - times ^ The type of administration and the frequency of administration;; the combination of any of the ways described in X here to form a drug administration cycle. Quxi Li> Negative Precursor, TCN, TCN-P and Phase The compound can be administered repeatedly through a certain administration cycle, for example, in a large dose, every two weeks - two months. The administration plan can be administered for at least a duration: 2, 3, 4, 5, 6, 7, 8 ' 9, 10, 1 Bu 12, 18, or 24 months. In addition, a patient may be administered at least 2, 3, 4, 5, 6, 7, 8, 9, 10, U, 12, 15 or 2 Dosing cycle. Tricineside prodrug, TCN, TCN-P and related compounds 56 200902034 According to any combination disclosed herein, for example, the drug can be administered once a week for three cycles every three weeks. In other embodiments, 'tricilide prodrug, TCN, TCN-oxime, and related compounds can be administered at least once a day for at least 2, 3, 4, 5, 6, 5, 7, 9, or 10 days. Such administration is followed by the corresponding cycle time of unadministered drug. As shown here, the tromethamine prodrug, TCN, TCN-Ρ and related compounds can be administered to the patient in an amount effective to cause tumor regression. Administration of TCN, TCN-oxime and related compounds may be provided in live 10, at least 15-20% of the individual at least partially reacted, If at least 15%, 20% or 30% reacts or completely reacts. In several embodiments, at least 2' 5, 10, 15, 20, 30 or 50 mg/m 2 of the triclinide compound disclosed herein can be administered To a body, in some embodiments, 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, 15 9.5 , 10, 12, 15, 17, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150 '165, 175, 200 250, 300, or 350 mg/m 2 As described herein, the triclinide prodrug, TCN, TCN-oxime, and related compounds can be administered to one body. Administration of the compound can be carried out according to any of the treatment plans disclosed herein. In a particular embodiment, the dosing schedule comprises administering a Tricinem prodrug, TCN, TCN-oxime, and related compounds of less than about 20 mg/m2 simultaneously, sequentially, or over a period of time. In one embodiment, the triclinide prodrug, TCN, TCN-oxime, and related compounds below 20 mg/m 2 can be administered once a week. In additional examples, 2 mg/m 2 , 5 mg/m 2 , 57 200902034 10 mg/m 2 and/or 15 mg/m 2 of triclinide prodrug, TCN, TCN-P and related compounds may Give a body. In another embodiment, less than 10 mg/m2 of triclinide prodrug, TCN, TCN-P, and related compounds can be administered to a body for at least five weeks by continuous infusion. 5 The present invention provides any combination of the type of administration, frequency, number of cycles, and dosage as disclosed herein. 6.1. Screening of Patient Populations In another aspect of the invention, methods are provided to identify cancers or tumors that are sensitive to the toxic effects of Tricinem (TCN) and related compounds. In one embodiment, a method for treating cancer or a tumor in a mammal is provided by (1) obtaining a biological sample from the tumor; (ii) determining whether the cancer or tumor overexpresses Akt kinase or highly activated and scalloped. Akt kinase, and (iii) treating the cancer or tumor with tripceine or a related compound as described herein. In one embodiment, the biological sample can be a biopsy. In other embodiments, the biological sample can be a fluid, cell, and/or aspirate derived from the tumor or cancer. Biological samples can be obtained according to any technique known to those skilled in the art. In one embodiment, a biopsy can be performed to obtain a biological sample. A biopsy is a procedure performed by the removal of tissue or cells from the body for examination. Such biopsy can be performed in a physician's office, but others may need to be performed in a hospital facility. In addition, some biopsy may require anesthesia to cause loss of consciousness in the area, while other tests do not require any anesthesia. In several embodiments, an endoscopic biopsy can be performed. This type of biopsy is passed through a fiberoptic endoscope (a slender 58 200902034 thin tube with a near (four) telescope at the end for viewing) through the self-supply orifice (also g-rectal) or a small incision (also That is, arthroscopy). The endoscope uses a problematic organ for abnormal or suspicious areas, and a small amount is available for research. Endoscopic surgery is named according to the organ or body area to be viewed and/or to be treated. The physician inserts the endoscope into the gastrointestinal tract (digestive tractoscopy), bladder (bladderoscopy), abdominal cavity (laparoscopic examination), joint cavity (arthroscopy), chest furnace; middle (longitudinal examination), or trachea With the bronchial system (pharyngoscopy and bronchoscopy). In another embodiment, a bone marrow biopsy is performed. This type of 10 biopsy can be performed by the sternum (sternum) or the sacral hip bone (the bone area on either side of the posterior pelvis). Clean the skin and give a local anesthetic to paralyze the area. The slender, rigid needle is inserted into the bone marrow and the cells are aspirated for study; this step occasionally causes discomfort. Central biopsy (removing small bone "shards" from the bone marrow) can be performed after the aspiration procedure. In another embodiment, excision or incision of a biopsy can be performed on a mammal. This plastic biopsy is often used when a wider or deeper skin portion is desired. Using a scalpel, remove the full skin thickness for further examination and suture the wound (using surgical sutures to suture off). When the entire tumor is removed, it is called a biopsy technique. When only a portion of the 20 tumor is removed, it is referred to as a biopsy technique. For example, when it is suspected to be a melanoma (a type of skin cancer), it is preferable to perform a biopsy. In still other embodiments, a fine needle aspiration (FNA) in vivo iron test can be used. This type of biopsy uses a fine needle to move the tumor out of the tiny pieces of tissue. Occasionally, local anesthesia is used to tickle the area, but this procedure is rare. 59 200902034 Major discomfort and no retention. For example, FNA is not used for the examination of suspected symptoms. 彳-叮 is used to loosen large lymph nodes near melanoma to understand whether melanoma has metastasized (diffused). Computed tomography (CT scan or CAT scan) can be used to introduce tumors into the internal organs such as the lungs or liver. In other embodiments, the punching, braking, and/or skin biopsy can be performed. Puncture biopsy involves the use of a biopsy instrument to remove a short cylinder or tissue called "Apple Heart" for deeper skin samples. After the local anesthesia is applied, the instrument is rotated on the surface of the skin until the instrument passes through the layers, including the dermis, epidermis, and the superficial portion of the skin (subcutaneous fat). Peeling the living group 1 The woven inspection involves removing the top layer of the skin by scraping. Puncture biopsy was also performed using local anesthesia. Skin biopsy involves removing the skin sample and examining it under the microscope to determine if melanoma is present. The biopsy is performed under local anesthesia. In a particular embodiment, a method of determining whether a tumor overexpresses Ak k kinase 15 is provided. The overexpression of Akt kinase can be referred to the phosphorylation state of the kinase. Acidification of Akt can be detected according to the methods described herein. In one embodiment, the tumor biopsy can be compared to control tissue. The control tissue may be a normal tissue from a mammal undergoing a biopsy or a normal tissue obtained from a healthy mammal. Akt stimuli overexpression or high phosphorylation can determine that tumor biopsies contain a greater amount of Akt kinase and/or Akt kinase phosphorylation than control tissues, such as at least about 1. 5, 2 than the Akt kinase contained in the control tissue. 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

Akt kinase. 200902034 In one embodiment, the invention provides a method of detecting a disordered octopus kinase expression in an individual or in a biological sample obtained from the individual, by administering cells, cell extracts, Serum or other sample or contact with the immune parent interaction molecule that is specific for the Akt kinase or antigenic portion thereof, and screening for the degree of formation of the immunological interaction molecule-Akt kinase complex, wherein the complex is present The amount is elevated relative to normal cells, indicating a disordered cell that can exhibit or overexpress Akt. In one example, immunocytology can be used to screen cells or cell extracts for the presence of elevated levels of Akt kinase. In another embodiment, the degree of expression of the Akt kinase gene is screened to detect the disordered expression of Akt in the cell at the genetic level, wherein the transcriptional expression product (ie, mRNA) is elevated in comparison with normal cells. Indicates out of order cells. In several embodiments, real-time PCR and other PCR programs can be used to determine transcriptional activity. In one embodiment, the mRNA can be obtained from 15 cells of the individual or from a biological sample obtained from the individual, and cDNA can be produced as needed. The mRNA or cDNA is then contacted with a gene probe that can hybridize to and/or amplify a full or partial nucleotide sequence encoding an Akt kinase or its complementary nucleotide sequence, and then detect the amount of mRNA or cDNA 'where the mRNA or mRNA can be assessed Whether the concentration of cDNA is higher than that of the normal control group. 2. Still another embodiment of the present invention encompasses an antibody against Akt kinase in a quantitative or semi-quantitative diagnostic kit set comprising a monoclonal antibody or a plurality of antibodies for determining Akt kinase in a suspected cancer cell derived from a patient. Relative concentrations, including all reagents required to perform the assay. In one embodiment, a kit of kits and materials required for performing an ELISA assay is provided. Test 61 200902034 agents may include, for example, washing buffer, antibody dilution buffer, blocking buffer, cell staining solution, developing solution, stopping solution, anti-phospho-protein specific antibody, anti-ubiquitin specific antibody, secondary antibody And distilled water. The kit group also includes instructions for use, either automated or semi-automated as needed, or in a form compatible with automated machines or software. In one embodiment, detection of the AKT live 4 匕 form (&amp; cyanoic acid 4740 acidified Akt) t stone H acid-ser-473 A kt antibody can be used as an antibody in the diagnostic kit set. For example, refer to Yu an et al. (2000), "The ovarian cancer frequently activates AKT2 and induces apoptosis via inhibition of the phosphinic acid-3-OH kinase/Akt pathway." Oncogene, 19: 10 2324-2330 . 6.2. Akt motivation

Akt is also known as PKB3, a subfamily of serine/threonine kinases. Three members AKT1, AKT2, and AKT3 have been identified in this subfamily. Akt is activated by extracellular stimulation in a PI3K-dependent manner (Datta S. R. et al. 15, Gene Dev, 13: 2905-2927, 1999). The full activation of Akt requires phosphorylation of Thr3()8 in the activation loop and Ser473 at the C-terminal activation site. Akt is negatively regulated by the PTEN tumor repressor. Mutations in PTEN have been identified in a variety of tumors, resulting in activation of the Akt pathway (Datta S. R. et al., Gene Dev, 13: 2905-2927, 1999). In addition, amplification, overexpression and/or activation of Akt has been detected in a variety of human 20 diseases (Datta SR et al, Gene Dev, 13: 2905-2927, 1999; Cheng J_Q. and Nicosia S_V., tumorigenesis) The AKT signal transduction pathway. For reference in Cancer Encyclopedia, Schwab D. (ed.), Berlin, Heidelberg and New York; Springer; 20CU, pp. 35-37). Akt specifically induces ectopic expression of active Akt, inducing cell survival and malignant transformation of 62 200902034; whereas inhibition of Akt activity induces apoptosis in mammalian cells (Datta SR et al., SgDev, 13: 2905- 2927, 1999; Cheng J_Q· and Nicosia SV, AKT signal transduction pathway for oncogenesis. Reference to Cancer Encyclopedia, Schwab D. (ed.), • 5 Berlin, Heidelberg and New York; Springer; 2001, pp. 35-37; Sun M. et al., Am J Path, 159: 431-437, 2001; Cheng J_Q. et al., Oncogene '14: 2793-2801 '1997). In addition, activation of Akt has been shown to be associated with tumor invasion and chemical resistance (West K.A. et al., Drug Resist Updat, 5: 234-248, 2002). Activation of the 10 Akt pathway plays a key role in malignant transformation and chemical resistance by inducing cell survival, growth, migration, and angiogenesis. The present invention provides methods for determining the extent of Akt kinase overexpression and/or the extent of hyperactivation and phosphorylation of Akt kinase.

The Akt kinase can be any of the known Akt family kinases, or their associated kinases, including but not limited to Aktl, Akt2, Akt3. The mRNA and amino acid sequences of human Aktl ' Akt2 and Akt3 are shown in Figures 6a-c, 7a-d and 8a-c, respectively. 6.3. Diagnostic assay 旮; bf immunoassay analysis 20 In one embodiment, a method for detecting the disordered expression of Akt kinase in a mammalian cell or in a biological sample obtained from a mammal is provided, which is obtained by The mammalian cell, cell extract or serum or other sample or biological sample is contacted with an immunological interaction molecule specific for Akt kinase or an antigenic portion thereof; and screening for an immunological interaction molecule 63 200902034 and determining whether or not there is a relative Increased amount of cell complex in normal-Akt kinase complex formation η 乂 interaction knife can be a specific molecule for the application of kinase or its antigenic part or its homologue or derivative. It can be an immunoglobulin molecule. In other embodiments, the immunologically interacting molecule can be an antibody fragment, a single-chain antibody, and/or a tau cell associated with an antigen-binding molecule. In a particular embodiment, the antibody can be It is a monoclonal antibody. In another specific implementation, the antibody may be a multi-strain antibody. The Qianshen molecule is specific to Shijisaki or more specifically to the antigenic stator or antigenic site on the Akt kinase. The antigenic stator or epitope on the Akt priming includes the molecular portion to which the immune response is directed. The antigenic stator or epitope may be a B cell epitope, or, if appropriate, a cellular antigenic site. In one embodiment The antibody is a phospho-ser 473 Akt antibody. One embodiment of the present invention provides a method for the presence of cancer growth or a series of cancer growth in which mammals have disordered Akt activity, and the cells are passed from the mammal. Or a cell extract or a biological sample obtained from an individual is bound to an Akt®|effective amount of an antibody having the specificity of the agonist or the antigenic stator or antigen thereof Site contact; and then quantitatively or sputum determination of the content of the Akt-stimulus-antibody complex, wherein the presence or absence of a higher level of the normal cell is determined. The k-body can be obtained by a variety of means known to those skilled in the art. Either one is prepared, for example, for the detection of human Akt kinase, which is typically, but not necessarily, derived from non-human animals such as primates, livestock animals (eg, sheep, cattle 1, goats, 64 200902034 horses), laboratory test animals (eg, Mouse, rat, guinea pig, rabbit) and/or accompanying animals (eg, dogs, cats). Antibodies can also be produced recombinantly in prokaryotic or eukaryotic host cells. In general, antibody-based assays can be performed. The test tube is carried out in a biopsy of cells or tissues. However, if the antibody is subjected to humanization by appropriate de-immunization or in the case of human use, the antibody may be labeled with a nuclear label, for example, and administered to a patient. Techniques for determining nuclear marker stacking positions. Akt kinase antibodies can be cancer targeting agents. Thus, another embodiment of the present invention is provided for use in humans and non-human patients. A deimmunized form of an antibody for cancer imaging. 10 In general, in order to produce an antibody against Akt kinase, the enzyme is required to be extracted from a biological sample, and the biological sample may be derived from an animal including human tissue, or may be manufactured by recombinant means. From cell culture. Akt kinase can be isolated from a biological sample by any suitable means. For example, isolation can utilize Akt kinase surface charge properties, size, density, biological activity, and binding to another entity (eg, Akt 15 kinase or other Any one or more of the affinities of another protein or chemical compound associated with the method. Thus, for example, isolation of Akt kinase by a biological fluid can be achieved by any one or more of the following methods: Centrifugation, ion-assisted tomography (eg, anion exchange chromatography, cation exchange chromatography), electrophoresis (eg, polyacrylamide gel electrophoresis, isoelectric poly 20 coke), size separation (eg gel filtration, super Filtration and affinity media separation (eg, immunoaffinity separation including, but not limited to, bead separation such as Dynabead (trade name) separation, Immunochromatography, immunoprecipitation). The separation of the Akt kinase from the biological fluid preserves the conformational epitope of the conformation present on the kinase&apos; so appropriately avoids the technique of causing the hair loss. In yet another embodiment 65 200902034, the agglutination can be separated from the biological fluid using any one or more of affinity separation, gel filtration, and/or ultrafiltration. Immunization and subsequent manufacture of monoclonal antibodies can be carried out using any of the standard protocol protocols known to the art, as described, for example, in Kohler and 5 Milstein (Kohler and Milstein, Nature, 256: 495-499, 1975; Kohler and Milstein, Eur J Immun〇1, 6(7): 511_519, 1976);

Coligan et al. (Popular Program for Immunology, John Wiley & Sons, 1991-1997) or Toyama et al. (Handbook for Individual Antibody Experiments, 1987, published by the Science Society). Generally, an animal is immunized by a standard method using a recombinant form of Akt kinase-containing biological fluid 10 or a portion thereof or Akt kinase to produce an antibody-producing cell, particularly an antibody-producing somatic cell (e.g., B lymphocyte). These cells are then separated from the immunized animal for immortalization. In several embodiments, Akt kinase fragments can be used to produce antibodies. The segment can be attached to a carrier. The carrier can be any of the typical high molecular weight materials of 15 Å, non-immunogenic substances or undesirable immunogenic substances (such as haptens) naturally linked or artificially linked to the carrier to enhance its immunogenicity. Immortalization of antibody-producing cells can be carried out using methods well known in the art. Immortalization can be achieved, for example, via the use of Epstein-Barr virus (EBV) transformation (Kozbor et al., Methods in Enzymology, 121: 14, 1986). In another 20 embodiment, antibody-producing cell lines are immortalized using cell fusion methods (described in Coligan et al., 1991-1997, supra), and cell fusion methods are widely used to produce monoclonal antibodies. In such a method, an antibody-producing somatic cell having the potential to produce an antibody, particularly a B cell line, is fused to a myeloma cell line. Such somatic cells may be derived from secondary infected animals, preferably rodents such as the lymph nodes, spleen and peripheral blood of rats and rats. In a particular embodiment, mouse spleen cells can be used. In other embodiments, rat, rabbit, sheep or goat cells can also be used. Specialized myeloma cells have been developed from lymphocytic tumors for use in fusion procedures for fusion tumor manufacturing (Kohler and Milstein, 5 1976, see above; Shulman et al., Nature, 276: 269-270'1978;

Volk et al, J Virol, 42(1): 220-227, 1982). A variety of myeloma cell lines can also be used in the manufacture of fusion cell hybrids, including, for example, P3 by 63-Ag8, P3 by 63-AG8.653, P3/NSl-Ag4-l (NS-1), Sp2/0-Agl4, and S194/5_XXO.Bu.l. The P3 by 63-Ag8 and NS-1 cell lines 10 have been described by Kohler and Milstein (1976, see above). Shulman et al. (1978 'see above) developed the Sp2/0-Agl4 myeloma cell line. The SI94/5.XXO.Bu.l cell line was reported by Trowbridge (J Exp Med, 148(1): 313-323, 1978). The method of producing antibody-producing spleen cells or a mixture of lymph node cells and myeloma cells is generally involved in the presence of an agent (chemical, viral or electrical) that promotes fusion of the cell membrane 15, and the somatic cells and the tumor cells are 10: 1 ratio mixing (but the ratio can vary from about 2 〇: 1 to about 1:1). Fusion methods have been described (Kohler and Milstein, 1975, supra; Kohler and Milstein, 1976, supra; Gefter et al, I* Cytogenetics, 3: 231-236, 1977; Volk et al., 1982, Springs See 20). The fusion promoters used by these researchers are Sendai virus and polyethylene glycol (PEG). In several embodiments, means are provided for selecting fusions, fine p-crosses derived from the remaining unfused cells, particularly unfused osteogenic tumor cells. In general, the selection of fused cell hybrids is accompanied by the culture of cells in the culture medium, which supports the growth of fusion tumors, and stops the growth of unfused myeloma cells. Will continue to split indefinitely. The somatic cells used for fusion are not cultured in vitro and do not retain long-term viability. This is not a problem. It takes several weeks to selectively culture the fusion, 'field hybrids'. Early in the day, it is necessary to identify such hybrids that produce the desired antibody 5, which can then be subsequently propagated and propagated. In general, about 1 〇〇/0 of the resulting hybrid can produce the desired antibody, but a range of from about 1% to about 30% is not uncommon. The detection of antibody-producing hybrids can be achieved by any of several standard assay methods, including enzyme-linked immunoassay techniques and radioimmunoassay techniques, such as those illustrated in 1^1111, etc. Strain and fusion tumors: a new era in bioanalysis, Plenum Press, New York, pp. 376-384, 1980; and can be reached by faCS analysis (O'Reilly et al., Biotechnology, 25: 824-830, 1998). Once the desired fused cell hybrid has been selected and transfected into individual antibody-producing cell lines, each cell line can be propagated in either of two standard ways. The fusion tumor cell suspension is injected into a tissue compatible animal. The injected animal is then developed to secrete specific monoclonal antibodies produced by the fused cell hybrid. Animal body fluids such as serum or ascites can be tapped to produce monoclonal antibodies at high concentrations. Alternatively, individual cell lines can be propagated in a laboratory culture vessel in a test tube. The culture 20 containing a high concentration of a single specific monoclonal antibody can be harvested by decantation, filtration or centrifugation, and subsequently purified. The cell line is then tested by immunoassay for its specificity for detecting the Akt kinase of interest. For example, the cell line can be distributed into a plurality of wells and cultured, and the supernatant from each well is analyzed by enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody technique. The individual antibody-producing cell line that recognizes the target LIM kinase 200902034 but does not recognize the non-target epitope is identified 'and then cultured directly in a test tube, or injected into a tissue-compatible animal to form a tumor, and The desired antibody is produced, collected, and purified. Accordingly, the present invention provides a method for detecting Akt kinase or a 5 segment thereof, a variant strain or a derivative thereof, which comprises contacting the sample with an antibody or a fragment thereof or a derivative thereof; and detecting a normal control group containing the antibody And a content of a complex of Akt kinase or a variant or derivative thereof, wherein the degree of elevation of Akt kinase is determined. Any suitable technique for determining the formation of a complex can be used. For example, an antibody 10 having a combined reporter molecule according to the present invention can be used in immunoassay analysis. Such immunoassay analysis includes, but is not limited to, radioimmunoassay analysis (RIA), enzyme-linked immunosorbent assay (ELISA), immunochromatography (ICT), and Western blotting. These immunoassays are well known to enthusiasts. . Immunoassay analysis also includes competitive assay analysis. The invention encompasses qualitative and quantitative immunoassay analysis. 15

Suitable immunoassay analysis techniques are described, for example, in U.S. Patent No. 4, (Π 6,043 424,279; and 4, 〇18,653. These techniques include non-competitive single-position and two-position assays and singularity assays. The assay comprises direct binding of the labeled antigen-binding molecule to the target antigen. The invention further provides a method for quantifying the degree of protein expression and degree of activation in an automated subject such as a cancer patient or a cell or tissue sample suspected of having a cancer. In one embodiment, the invention provides a method for quantifying the extent of Akt protein expression or activation using an imaging system. The imaging system can be used to receive, enhance, and process cell samples or tissues that have been stained with Ακτ protein specific 69 200902034 staining. An image of the sample, which is determined by the amount or degree of activation of the AKT protein in a cell or tissue sample obtained from such an animal. In one embodiment of the method of the invention, at least two of the AKT proteins exhibiting unequal amounts of AKT protein The cell line produces a calibration curve of AKT 1 and AKT2 protein 5 quality. The curve is used to quantify the amount of AKT protein expressed in a cell or tissue sample. An activation characteristic-specific reagent can be used to make a similar calibration curve for the activated AKT protein. It can also be used to determine the Ακτ before and after treatment of clinical cancer. A number and a change in the Ακτ activation state. 10 In a particular embodiment of the method of the invention, the degree of expression of the A Κ Τ protein in a cell or tissue sample can be determined by enzyme-linked immunosorbent assay (ELISA) to determine the sample. Ακτ protein content. Such a method is described, for example, in U.S. Patent Publication No. 2〇〇2/〇〇丨5974. In other cases, it can be detected in the analysis of the Akt5 enzyme in this assay, rugged Incorporation with a second antibody, usually by glutarazine or a moth molysate, to a second antibody. The enzyme substrate for a particular enzyme is typically selected to produce a detectable color change upon hydrolysis by the corresponding enzyme. It is also possible to use a light-producing enzyme substrate that obtains a fluorescent product instead of using a chromogenic enzyme substrate. The enzyme-labeled antibody can be added to the first antibody-anti-2〇. The complex is allowed to bind and then washed to remove excess reagent. The solution containing the appropriate enzyme matrix is then added to the antibody-antigen-antibody complex. The enzyme matrix reacts with the binding antibody to the second antibody to generate a quantitative visual signal. Tongshi can use the spectrophotometric method to obtain the indication of the amount of antigen present in the sample towel. In addition, fluorescent compounds such as luciferin, rhodamine, 70 200902034 and lanthanide compounds, pin (EU) can be used. The chemistry of the king is a few bodies without changing its binding ability. When the light is illuminated by a specific wavelength of light, the fluorescently labeled antibody absorbs light energy, induces excitation in the molecule, and then uses it frequently. The optical microscope can visually detect the victory of 6 * hearts will illuminate the color of light. The Lloyd-labeled antibody is allowed to bind to the first anti-髀γ κ &gt; 赗-antigen set. The remaining ternary (qua) (4) family complex is exposed to light of the appropriate wavelength after washing to remove unbound reactants. Observed (4) refers to the antigen of interest in money. The immunofluorescence metrological assay (A) can be used in this method in the “Boundary”, but other notification sub-segments, such as radioisotopes, chemiluminescent molecules or bioluminescent molecules, can also be used. In the case, antibodies to Akt akisaki can also be used for detection of Akt kinase by elisa media in sera or other circulating body fluids. The anti-Akt kinase antibody can be immobilized on a solid target and allowed to be subjected to a biological extract such as Serum, blood, lymph or other body fluids, cell extracts 15 or cell biopsy contacts are achieved, and labeled anti-purine kinase antibodies can be used to detect _braking Akt_. This assay can be performed in a variety of ways, all Variations are encompassed by the present invention and are known to those skilled in the art. This approach allows for the rapid detection and quantification of Akt kinase levels, for example, using serum-based assays. 20 In one embodiment the Akt ELISA assay kit kit can be used in this embodiment. The invention may be used, for example, in a cell-activated elisa kit for Akt S473 from SuperArray Bioscience. In one embodiment, the antibody can be an anti-pan-antibody that recognizes Akt S473. The ELISA assay kit kit contains anti-Akt antibodies and additional reagents, including but not limited to 71 200902034 in wash buffer, antibody dilution buffer, A buffer solution, a cell staining solution, a developing solution, a stop solution, a secondary antibody, and distilled water. Nucleotide detection is provided in another embodiment to provide a degree of expression in a cell by detecting a polynuclear nucleoside encoding Akt. Methods for detecting Akt kinase. The expression of a polynucleotide can be determined using any suitable technique known to those skilled in the art. In one embodiment, a labeled polynucleotide encoding an Akt kinase can be used as a probe for obtaining from a cell. Northern blot analysis of the RNA extract. In other embodiments, the nucleic acid extract of the animal may be associated with a message sequence and an anti-message sequence of a polynucleotide encoding the kinase 10 or a collateral sequence thereof. Oligonucleotide primers are used in synergistically for nucleic acid amplification reactions, such as RT PCR. A variety of automated solid phase detection techniques are also available to those skilled in the art. For example, it is described in Fodor et al. (Science, 251: 767-777, 1991) and Kazal et al. (Nature Drugs, 2: 753-759, 1996). 15 In other embodiments, detection of RNA encoding Akt kinase is provided. A method of transcript. RNA can be isolated from a sample of cells suspected of containing Akt kinase rna, such as total RNA isolated from human cancer tissue. RNA can be obtained by methods known in the art, for example using TRIZ0L reagent (GIBC0-BRL/Life Sciences, Maryland) Separation of oligo-dT or random sequence oligonucleotides and specific sequence oligonuclear 2 nucleosides can be used as primers in the reverse enzymatic reaction to prepare the first strand of cDNA obtained from the isolated RNA. The resulting first stock is then amplified in a pCR reaction with a specific sequence of oligonucleotide oxalate to obtain an amplification product. Polymerase chain reaction or &quot;PCR&quot; refers to a procedure or technique in which the amount of nucleic acid, 11&lt;/RTI&gt;, and/or preselected sputum fragments of DNA is amplified, for example, 72 200902034 to U.S. Patent 4,683,195. In general, sequence information from the end of the region of interest or beyond the region is used to design oligonucleotide citrate primers. The sequence of such primers may be the same or similar to the sequence of the strands to be amplified. PCR can be used to amplify specific RNA sequences and cDNA transcribed from whole cell RNA. Large reference is made to Mullis et al. (Quant Biol, 51: 263, 1987; Erlich, ed., PCR, Starkton, New York, ι 989). Thus, amplification of a particular nucleic acid sequence by PCR relies on an oligonucleotide or primer having a retained nucleotide sequence that is presumed by the alignment of the relevant gene sequence or protein sequence, such as by the sequence of a mammalian Akt kinase gene. Compare 10 presumptions. For example, a primer is prepared which is predicted to be paired with an anti-message strand and another primer is predicted which is predicted to be paired with a message strand encoding a cDNA molecule of Akt kinase. To detect the amplified product, the reaction mixture is typically subjected to agarose gel electrophoresis or other convenient separation technique to detect the relative presence of Akt kinase-specific amplified DNA. For example, 15 Akt kinase-amplified DNA can be detected by Southern hybridization with a specific oligonucleotide probe, or its electrophoretic activity can be compared to a DNA standard of known molecular weight. Isolation, purification and characterization of the amplified Akt kinase DNA can be performed by excising the fragment from the gel or eluting the fragment (for example, reference to Lawn et al, Nucleic Acids Res., 2: 6103, 1981; Goeddel et al. Nuclear 20 Acid Research, 8: 4〇57, 1980); the amplified product is transferred into a suitable vector such as the pCRII vector (jnvitr〇gen); Subsequence; and the DNA sequence is achieved by comparison with known sequences of LIM kinase. The relative amount of LIM kinase mRNA and cDNA was then determined. 73 200902034 In one embodiment, real-time PCR is used to determine the degree of transcription of Akt nucleotides. The determination of transcriptional activity also includes assays based on the possible translational activity of available mRNA transcripts. The real-time PCR program and other PCR programs use a variety of PCR products for detection chemistry, including binding of dna-binding fluorophores, 5, 5 endonucleases, adjacent linings and hairpin oligo probes, and self-fluorescence amplification. gene. These chemical and real-time PCRs are discussed in general by Mackay et al., Nucleic Acids Research, 30(6): 1292-1305, 2002; Walker, J Biochem Mol Toxicology, 15(3): 121-127, 2001; Lewis Et al., J Pathol, 195: 66-71, 2001 〇10 In another embodiment, the disordered expression of Akt can be identified by allowing a nucleotide sequence isolated from a biological sample to have a Contacting the Akt sequence of the oligonucleotide sequence of Figure 6a-c, 7a-d, or 8a-c, or the nucleotide probe of one of the flanking and complementing sequences; and then via the probe The parent compares the result with a normal sample to detect the sequence. The parent of the biological sample can be detected by labeling the probe with any one of the detectable agents. The probe may, for example, be a radioisotope or be labeled with biotin, a fluorescent dye, an electron dense reagent, an enzyme, a hapten or a protein which can be obtained. The detectable label can be assayed by any desired means including spectroscopic means, photochemical means, biochemical means, immunochemical hand &amp; radioactive 20 isotope means, or chemical means. Probes can also be used, for example, in oligo-acid oxalate limiting techniques, dot-point assays, inversion-point assays, line probe assays, and 5, Wei enzyme assays. In addition, the probes can be detected using the most suitable surface array technology, including giant array technology, microarray technology and germanium microchip technology. Oligonucleotide probes typically 74 200902034 include about 14, 15, 16, 18, 20, 25 or 28 hybridized to nucleotides selected from the 6a-c, 7a-d and 8a-c or fragments thereof Nucleotide. Probes having a length greater than about 25 or 28 nucleotides are generally not preferred. Oligonucleotide probes are designed to recognize Akt nucleotide sequences. 5 Kinase assay analysis

The activity of Akt kinase can be determined using any of the appropriate kinase assays known to the art. For example and without limitation, Hogg et al. (oncogene, 1994.9: 98-96), Mills et al. (J Biol Chem, 1992. 267: 16000-006) and Tomizawa et al. (FEBS Lett, 2001, 492: 10 221) can be used. -7) 'Method described by Schmandt et al. (J Immun〇i, 1994. 152: 96-105). Additional assays for serine, threonine, and tyrosine kinase assays are described in Ausubel et al. (Short Protocol Methodology in Molecular Biology, 1999, Unit 17.6).

Akt kinase assays typically employ an Akt polypeptide, a labeled donor enzyme 15 matrix, and a receptor enzyme matrix that is specific or non-specific for Akt. In this assay, Akt transfers the labeled portion from the donor enzyme substrate to the 5:body enzyme matrix, and the kinase activity is determined by the amount of the δ-hex moiety transferred from the donor enzyme substrate to the receptor enzyme substrate. Akt polypeptides can be produced using a variety of expression systems, can be purified from cells, can be in the form of 20 fused proteins that are lysed or uncleaved, and/or can have non-鸠 polymorphic sequences such as celestial or beta-galactose At its N end red end. If the secret cell (4) is the source of the Akt to be assayed, the Akt activity can be assayed in a cancerous cell line. Appropriate application of the enzyme enzyme substrate for analysis includes any sensation that is sensitive to descaling, for example, including the 丫 _ mark and the class 75 200902034, wherein the label is 33P, 32P, 35S or Any other radioisotope or appropriate fluorescent label. The appropriate recipient enzyme matrix for Akt assay analysis includes any polypeptide or other molecule that is sensitive to phosphorylation by Akt. The recipient enzyme matrix is derived from an in vivo Akt target fragment. The acceptor enzyme substrate fragment 5 can be from 8 to 50 amino acids in length, typically from 1 to 30 amino acids, particularly about 10, 12, 15 '18, 20 and 25 amino acids. Other recipient enzyme matrices can be assayed experimentally using a different set of polypeptides or other molecules. The receptor substrate target of TTK can be purified from other reaction components once the reaction is carried out. This purification is usually carried out via molecular interaction, where the recipient enzyme 10 is biotinylated and purified by interaction with Streptavidin; or a specific antibody can be obtained, the antibody can be specific Sexual recognition of the recipient enzyme matrix. The reaction is carried out under a variety of conditions, such as on a solid support, in a gel, in solution or in living cells. The choice of detection method is based on the labeling of the donor molecule. The method of use includes, for example, measurement of bound radioactivity or fluorescence by 15 radiography, scintillation counting, scanning, or fluoroscopy. 7. Methods of Treatment The compounds and pharmaceutical compositions provided herein are useful in the treatment of conditions including tumors and other disorders associated with abnormal cell proliferation. In one embodiment, the compounds of the invention are useful in the treatment of carcinoma, sarcoma, lymphoma, leukemia, and/or myeloma. In other embodiments of the invention, the compounds disclosed herein are useful for treating solid tumors. The compounds of the invention are useful in the treatment of cancer, such as, but not limited to, cancers of the following organs or tissues: breast, prostate, lung, bronchi, large intestine 'Breaking 76 200902034 urethra, bladder, non-Hodgkin's lymphoma, melanoma, Kidney, adrenal gland, pancreas, pharynx, thyroid, stomach, brain, multiple myeloma, esophagus, liver, intrahepatic bile duct, cervix, larynx, acute myeloid leukemia, chronic myelogenous leukemia, soft tissue such as heart, Hodgkin's Lymphoma, testicular, small intestine, chronic myelogenous leukemia, acute lymphocytic leukemia, anal, anal canal, anorectal, thyroid, genital, gallbladder, pleura, eye, nose, nasal cavity, middle; nasopharynx, ureter, abdominal cramps, Mesenteric, mesenteric, and gastrointestinal tract, high-grade gangeatoma, glioblastoma, large intestine, rectum, pancreas, gastric cancer, hepatocellular carcinoma; head and neck cancer, carcinoma; renal cell carcinoma; renal cancer; sarcoma; 15 20 Endothelial; lymphoma; leukemia, mycosis fungoides. In additional embodiments, the compounds of the invention are useful in the treatment of dermatological conditions including, but not limited to, malignant hemangioma, blood &amp; endothelial, basal cell carcinoma, squamous cell carcinoma, malignant melanoma, and card, West's sarcoma; And non-malignant or non-malignant conditions such as cognac, lymphangiogenesis, hemangioma in children, Sturge-Weber syndrome, night, neurofibromatosis, tuberous sclerosis, abdominal granulation Swollen, recessive dystrophic macrobubble epidermolysis, venous pain, hemorrhoids, rosacea, moist therapy, infectious soft night, lipid leakage keratosis' and linear keratosis. Compositions comprising a compound of the invention are useful for treating such cancers and other cancers from any stage of the discovery of cancer to the stage of ignorance. Furthermore, compositions comprising the compounds of the invention are useful in the treatment of primary cancers and metastatic cancers. In other embodiments of the lacquer lamp, the compounds described herein are useful for treating cancer, including but not limited to the cancers listed in Table 1 below. 77 25 200902034 □ Table 1: Acute lymphoblastic leukemia of the cancer category, acute lymphoblastic leukemia in adults, acute myeloid leukemia in children, acute myeloid leukemia in adults, adrenal cortical carcinoma in children, adrenal cortical carcinoma in children, AIDS-related cancer AIDS in children Related lymphoma anal cancer astrocytoma, childhood cerebellar stellate cell tumor, childhood brain basal cell carcinoma cholangiocarcinoma, extrahepatic cholangiocarcinoma, childhood bone cancer osteosarcoma / malignant fibrous histiocytoma brain stem neurotroph , childhood brain tumor, adult brain tumor, brain stem neuroglioma, childhood brain tumor, cerebellar stellate cell tumor, childhood brain tumor, brain astrocytoma / malignant glioma, childhood brain tumor, ependymoma, child Brain tumor, medulloblastoma, childhood brain tumor, undifferentiated neuroectodermal tumor on the cerebellum, childhood brain tumor, visual pathway and hypothalamic neurofibroma, childhood brain tumor, childhood breast cancer, childhood breast cancer, male Bronchial adenoma/carcinoid, child Burkitt's lymphoma, carcinoid tumor, gastrointestinal tract Unknown primary central nervous system cancer lymphoma, primary cerebellum, stellate cell tumor, childhood brain, stellate cell tumor/malignant glioma, child&quot; cervical cancer childhood cancer □ hair cell leukemia head and neck cancer Hepatocyte (liver) cancer, adult (primary) hepatocyte (liver) cancer, child (primary) Hodgkin's lymphoma, adult Hodgkin's lymphoma, child Hodgkin's lymphoma, pregnancy Hypopharyngeal hypothalamic and visual pathway neuroglioma, children with intraocular melanoma and pancreatic islet cell carcinoma (endocrine pancreas) □ Kaposi's sarcoma (kidney cell) cancer, renal cancer, 'children', laryngeal cancer Laryngeal cancer, childhood leukemia, acute lymphoblastic, adult leukemia, acute lymphoblastic, childhood leukemia, acute myeloid, adult leukemia, acute bone riding, childhood leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, B cell lip and oral cancer liver cancer, adult (primary) liver cancer, children (primary) liver cancer, non-small cell liver cancer, small cell lymphoma, AIDS-associated lymphoma, cypress Lymphoma lymphoma 'skin T cells, reference to sputum granuloma and Sezary syndrome lymphoma, Hodgkin's, adult lymphoma, Hodgkin's, childhood lymphoma 'He Jie Jin's, Pregnancy lymphoma 'non-Hodgkin's, adult lymphoma, non-Hodgkin's, childhood lymphoma' non-Hodgkin's, gestational lymphoma 'primary central nervous system □ macroglobulinemia, Wo Waldenstrom's malignant fibrous histiocytoma / osteosarcoma medulloblastoma, childhood melanoma melanoma, intraocular (eye) 78 200902034 Chronic lymphocytic leukemia Chronic myelogenous leukemia Chronic myeloproliferative disorders Colorectal cancer colorectal cancer, children's skin T-cell lymphoma, reference verrucous granuloma and Xie Saray syndrome □ endometrial cancer ependymoma, children's esophageal cancer esophageal cancer, children's Ewen's tumor family cranial embryo Cell tumor, childhood gonadal blast cell tumor, extrahepatic cholangiocarcinoma, eye cancer, intraocular melanoma, cerebral cancer, retinoblastoma, gallbladder, gastric cancer, gastric cancer, gastrointestinal carcinoid Tumor blast cell tumor, Gu Wai, childhood blast cell tumor, gonadal blast cell tumor, ovarian surviving trophoblastic tumor neuroglioma, adult glioma, childhood brain stem neuroglioma, childhood brain astrocytoma neuroglioma , children's visual path and hypothalamus skin cancer (melanoma) skin cancer, silent cell small cell lung cancer small intestine cancer soft tissue sarcoma, adult soft tissue sarcoma, childhood squamous cell carcinoma 'reference skin cancer (non-melanoma) There are hidden primary squamous cell carcinoma, metastatic gastric cancer, 1 child with undifferentiated neuroectodermal tumor on the cerebellum, children's □ T-cell lymphoma, skin, reference to sputum sputum swell and Xie Saray syndrome (Merkel) cells mesothelioma, adult malignant mesothelioma, children with multiple metastatic squamous cell carcinoma, multiple endocrine neoplasia syndrome, children with multiple osteosarcoma/plasma tumor verrucous granulation Bone marrow dysplasia syndrome bone marrow dysplasia / myeloproliferative myeloid leukemia, chronic myelogenous leukemia, adult acute myeloid leukemia, children Childhood acute myeloma, multiple myeloproliferative disorders, chronic nasopharyngeal and paranasal sinus cancer, nasopharyngeal carcinoma, nasopharyngeal carcinoma, childhood neuroblastoma, non-Hodgkin's lymphoma, adult non-Hodgkin's lymphoma, non-children Jajin's lymphoma, non-small cell lung cancer in pregnancy □ oral cancer, oral cancer in children, lip and oropharyngeal cancer osteosarcoma/malignant fibrous bone histiocytoma ovarian cancer, ovarian epithelial cancer ovarian blast cell tumor ovarian low malignant potential Tumors, cancer, pancreatic cancer, childhood pancreatic cancer, islet cells, paranasal sinus and nasal cancer, thyroid cancer, penile cancer, pheochromocytoma, pineal blastoma, and cerebellar undifferentiated neuroectodermal tumor, cerebral sinusoidal tumor cells Tumor/multiple bone thymoma pleural pulmonary blastoma pregnancy and breast cancer pregnancy and Hodgkin's lymphoma pregnancy and non-Hodgkin's lymphoma I ▲ hair, lower central nervous system lymphoma 79 200902034 睪9 cancer thymoma, Children with thymoma and thymic carcinoma, squamous adenocarcinoma, squamous adenocarcinoma, transitional cell carcinoma trophoblastic tumor of children with renal pelvis and ureter, surnamed □ unknown original location, Adult unknown origin, cancer, abnormal cancer of children and children, ureter and renal pelvis, transitional cell carcinoma, urethral cancer, uterine cancer, endometrial uterine sarcoma, vaginal cancer visual path and hypothalamic ganglia, children with genital cancer □ Wharden's giant globulin virus disease Wilms' tumor prostate cancer □ rectal cancer kidney cell (kidney) cancer kidney cell (kidney) cancer, children's pelvis and ureter, transitional cell carcinoma retinoblastoma Rhabdomyosarcoma, Child □ Salivary Gland Salivary Salivary Adenocarcinoma, Childal Sarcoma, Owen's Tumor Family Sarcoma, Kaposi's Sarcoma, Soft Tissue, Adult Sarcoma, Soft Tissue, Child's Sarcoma, Uterus Xie Saray Syndrome Skin Cancer (Non Melanoma) Skin Cancer , Children In other embodiments of the invention, the compounds disclosed herein are useful for treating diseases associated with angiogenesis. Anti-angiogenic small molecules include thalidomide, part 5 of which acts through inhibition of NFkB; 2-methoxyestradiol affects microtubule formation and activation of hypoxia-inducible factor (HIFla); Enzyme 2 (COX2) inhibitors; and low doses of conventional chemotherapeutic agents include cyclophosphamide 'taxanes' and vinca alkaloids (vincristine, vinblastine) )) (D'Amato RJ et al. (1994), Proc. Natl. 10 Acad. Sci. USA, 91, 3964-3968; D'Amato RJ et al. (1994), Proc. Natl. Acad. Sci. USA, 91,4082-4085). In addition, some serotonin inhibitors indirectly reduce angiogenesis by reducing the production of VEGF and other pro-angiogenic factors by tumors and stromal cells. These drugs include Herceptin, imatinib 80 200902034 (Glivec) and iressa (Bergers q et al. (2003), J Clm Invest, 11 1287-1295; Ciardiello F. et al. (2001), Clinical Cancer Research, 7, 1459-1465; Plum SM et al. (2003), Clinical Cancer Research, 9, 4619-4626). 5 late near 'angiogenesis inhibitors have been moved from animal research models to human patient studies. Angiogenesis inhibitors represent a promising therapeutic approach for a variety of different cancers. Recently, Avastin is an antibody with high affinity for vascular endothelial growth factor (VEGF). Avastin shows that renal cell carcinoma for deterioration can be used as a single agent to prolong life and be used for the deterioration of the large intestine. Cancer-binding chemotherapy can prolong life (Yang JC et al. (2003), New England Journal 349, 427-434; Kabbinavar F. et al. (2003), Journal of Clinical Oncology 21, 60-65). Tuberculosis-related diseases include, but are not limited to, inflammatory diseases, autoimmune diseases, and infectious diseases; angiogenesis-dependent cancers include, for example, solid tumors, blood 15 system tumors such as leukemia and tumor metastasis; benign tumors such as hemangiomas, sputum tumors, Neurofibromatosis, tracheal tumor, and pyogenic granuloma; wind, arthritis; dryness; wet therapy; ocular neovascularization such as diabetic retinopathy, premature retinopathy, macular degeneration, corneal transplant rejection , neovascular glaucoma, posterior lens hyperplasia, iris red syndrome; Oslo-Weibo (0sler_Webber) syndrome; myocardial angiogenesis; plaque neovascularization; capillary ash tube dilatation; bloodthirsty joint; And wound granuloma. Furthermore, the compositions of the present invention are useful in the treatment of diseases such as, but not limited to, intestinal adhesion, atherosclerosis, scleroderma, nocturnal and hypertrophic lesions (i.e., neoplasms). The composition of the present invention can also be used for the treatment of angiogenesis due to pathology 81 200902034, such as cat scratching heat (Rochele minalia quintosa), ulcer (Helobacter pylori) , tuberculosis and lupus. 7.1 Drug Resistance Tumor Cancer 5 The present invention provides compounds useful for the treatment of drug resistant cancer, including embodiments of the cancers and medicaments disclosed herein. In one embodiment, the compound as disclosed herein, such as triclinide prodrug, TCN, TCN-P, and related compounds, can be co-administered with a second drug. Multiple drug resistance (MDR) occurs in human cancer and may be a major obstacle to the success of chemotherapy 10 . Multidrug resistance is a phenomenon in which a tumor cell is exposed to a cytotoxic agent in a test tube to develop a chemically and chemically resistant range of compounds that are structurally and functionally related. In addition, multidrug resistant MDR may also occur uniquely in certain cancers without prior exposure to chemotherapeutic agents. Thus, in one embodiment, the invention provides a method for treating a patient suffering from a drug-resistant cancer, such as a multi-drug resistant cancer, by administering a Tricinem prodrug, TCN, TCN-P, and related compounds as disclosed herein. TCN, TCN-P and

Know or the cancer described here. law. In several embodiments, the triclinide prodrug-related compound can be used to treat paclitaxel, phenanthrene, cepacidine, and/or gefitini (Arisa) alone. 8. Combination therapy 82 200902034 In one aspect of the invention, the compounds and compositions disclosed herein can be administered in combination with at least one additional chemotherapeutic agent. Such additional agents may be administered in combination or alternation with the compounds disclosed herein. The drugs may constitute a part of the same composition, or may be administered as separate compositions 5 at the same time or at different times. In one embodiment, the compounds disclosed herein can be combined with anti-angiogenic I to enhance their effect, or combined with other anti-angiogenic agents and administered together with other cytotoxic agents. In another embodiment, the compounds and compositions, when used in the treatment of solid tumors, are selected from the group consisting of, but not limited to, the following agents: vitamins, vitamins, interferons, angiostatin, endostatin. , Shaly sinensis, thrombospondin-1, thromboxane-2, capt〇pryl, antitumor agents such as interferon, c〇Mp (cyclophosphamide, wenclidin , methotrexate and prednis〇ne), etoposide, mBACOD (methylamine oxime, bleomycin, doxorubicin, Cyclophosphamide, wendysone and dexamethasone, PRO-MACE/MOPP (Punisone, amidoxime (w/leucovin rescue), Dorothy Bixin, cyclophosphamide, mechlorethamine, wennedin, prenisson and procarbazine, wen christian, vinbrastine , angiostatin, TNP-470, pentosan polysulfate, platelet factor 4, angiostatin, LM-609, SU-1 (Π, CM-1 (Π,泰嘉兰(Techga) Lan), salivir, SP-PG, and radiation. In additional embodiments, the compounds and compositions disclosed herein can be administered in combination or alternately with the following drugs: for example, having an anti-silver 83 200902034 splitting effect. Drugs such as standard cytoskeletal elements, including podophyllotoxin or vinca bioassay (Wen Christian, von Brastin); antimetabolites (such as 5-tank π set, cytarabine) , gemcitabine, guanidine analogues such as pentostatin, 曱5 amine oxime); alkylating agents or nitrogen mustard gas (such as nitrosourea, cyclophosphamide, or ifosphamide) )); DNA-based drugs such as the antracycline drug adriamycin, doxorubicin, pharmorubicin or epirubicin; Drugs that are determined by topoisomerase such as etatopse; hormones and hormone agonists or 10 hormone antagonists such as estrogens, antiestrogens (tamoxifen and related compounds) and androgens, Flutamide, leuprorelin, ge G0sereiin, cyprotr〇ne, or octreotide; drugs that target signal transduction in tumor cells, including antibody derivatives such as Hepatic; burnt drugs such as Drugs (West 15 platinum, carbonplatin, oxaiiplatin, Φ white pluck; paraplatin) or subnutrazine; drugs that may affect tumor metastasis, such as matrix Metalloproteinase inhibitors; gene therapy and anti-information agents; antibody therapy; among which marine-derived bioactive compounds are notable for didemnins such as aplidine; steroid analogues 2, especially for Desa Mesa Pine; anti-inflammatory drugs including non-steroidal agents (such as acetaminophen or ibuprofen) or steroids and their derivatives, especially tesamethoxas; antiemetics including 5HT_3 inhibitors (such as gram beige (gramisetron) or Panta (壮ndasetr〇n)) and steroids and their derivatives are especially Desamethasone. In still other embodiments, the composition and composition may be administered in combination or alternately with the chemotherapeutic agents disclosed in Table 2 below.

Chemotherapeutic agents Table 2:

_13_cis-vitamin acid-2-amino-6-mercaptopurine-2-CdA-2-deoxyadenosine-5-fluorourine bite-5-FU -6-TG -6-bird嘌呤-6-疏基嘌吟-6-MP -Accutane - Actinomycin-D - Adriamycin - Adrucil - Agrylin ) - Ala-Cort - Aldesl eukin - Alemtuzumab - Alitretinoin - Alkaban - AQ • Aklan (Alkeran) ) - All-trans retinoic acid-α interferon - Altretamine - Amethopterin - Amifostine - Aminoglutethimide - Anage Anagrelide - Anandron - Anastrozole

- arabinose-Ara-C - Aranesp - Aredia - Arimidex - Aromasin

- Arsenic trioxide - Aspartate - ATRA - Avastin _ - Neosar · Neulasta - Neumega Neupogen - Neland Nilandron - Nilutamide - Nitrogen mustard - No valdex - No vantrone • Octreotide - Acetate Occidental - Oncospar ) - Oncovin - Ontak Onxal - Oprevelkin - Orapred • Orasone - Oxaliplatin - Paclitaxel - Pamidronate - Panretin - Paraplatin - Pediapred - PEG Interferon - Pegaspargase - Pegasus Pegfilgrastim - PEG-INTRON -PEG-L-aspartate - phenylmalonic acid mustard - Platinol - Platino - AQ - Prednisolone - Prednisone - Prelone - Procarbazine - PROCRIT - Proleukin - I with Carmustine implants I Liverpool (Prolifeprospan) ) 20 85 200902034

-BCG

-BCNU - Bevacizumab - Bexarotene

-Bicalutamide -BiCNU -Blenoxane -Bleomycin -Bortezomib -Busulfan -Busulfex -C225 -Leucovorin #5 -Campath - Camptosar - Camptothecin - l 1 • Capecitabine - Carac - Carpenter Carboplatin - Carmustine - Caso Ding - Casodex

-CCNU

-CDDP -CeeNU -Cerubidine •cetuximab -Chlorambucil -Cisplatin -Citrovorum Factor -Kola Cladribine - Cortisone Cosmegen - CPT-11 - Cyclophosphamide - Cytadren - Cytarabine - 赛塔拉宾Liposomes _ Cytosar-U - Cytoxan • Dacarbazine - Dactinomycin - Darbepoetin a - Daunorubicin (Darino) Daunomycin) • 'Daunorubicin' Daunoubi (DaunoXome) _ - Purinethol - Raloxifene - Rheumatrex • Ruitu Mountain (Rituxan) - Rituximab - Roveron - A (interferon a-2a) - Rubex - Rubidomycin - Santos Stading Sandostatin)

-Sanduo Stade LAR - Sargramostim - Solu-Cortef - S〇lU-Medrof - STI-571 - Streito Zuo Xin ( Streptozocin) - Tamoxifen - Targretin - Taxol - Taxotere - Temodar ''Temozolomide - Tunippi Teniposide

-TESPA - Thalidomide - Thalomid - TheraCys - Thioguanine - Thioguantoin - Thiophosphoamide • Thioplex - Thiotepa

-TICE - Toposar - Topotecan - Toremifene - Trastuzumab • Tretinoin - Trexall - Cui Senno (Trisenox)

-TSPA

-VCR -Velban -Velcade -VePesid -Vesanoid - Viadur -Vinblastine 86 200902034 -Deka Left (Decadron) - Vincasar Pfs - Delta-Cortef - Vincentine - Deltasone - Vinorelbine - Dai Nirukin (Denileukin) Dytoto-diftitox -VLB - DepoCyt - VP-16 - Dexamethasone - Vumon - Desa Mesa Pine - Like Xeloda - Zanosar - Desasone - Zevalin - Dexrazoxane - Zinecard - DHAD -Zoladex -DIC -Zoledronic acid -Diodex -Zometa -Docetaxel -Gliadel paste-mat Doxil) - Glivec - Doxorubicin - GM-CSF - Doxorubicin - Goserelin - Droxia - Granular cells - Community Stimulating Factor - DTIC - Granulocyte - Macrophage Cell-Community Stimulating Factor-DTIC-Dome-Hlotestin-Duralone-Herceptin-EfUdex-Hexadrol- Eligard - Hexalen - Ellens - Eloxatin - HMM - Elspar - Hycamtin -Emcyt -Hydrea -Epimbicin -Hydrocort -Epoetin a -Hydrocortisone - Erbitux (Erbitux) - hydroxycortisone sodium - Erwinia L-aspartame-sodium xycodone sodium-Estramustine - acid ketone ( Hydrocortone) - Ethyol - Hydroxyurea - Etopophos - Ibritumomab - Etoposide - Tiuxetan - Phosphate Ito Idamycin - Eul exin - Idarubicin - Evista - Ifex - Yi; Exemestane - IFN- a -Fareston - Ifosfamide ) - Faslodex - IL-2 - Femara - IL-11 - Filgrastim - Imatinib - Floxuri Din) -Imidazole Carboxamide -Fludara -Interferon alpha -Fudadarbin -Interferon a-2b (PEG conjugate) 87 200902034

-Fluoroplex - urinary feeding - fluorouracil (cream) - Fluoxymesterone - Flutamide - Folinic Acid - FUDR - Fu Fulvestrant -G-CSF •Gefitinib -Gemcitabine - Gemtuzumab Ozogamicin - Gemzar - Ge Gleevec - Lupron - Lupron Depot - Matulane - Maxidex - Mechlorethamine - Mecorosamin Hydrochloride -Medralone - Medrol 'Megace' - Megestrol -Melphalan - Mesna - Mesna Mesnex - Methotrexate - Methylprednisolone - Mylocel - Letrozole - Interleukin-2 - Interleukin-11 - Intron A (Interferon a-2b) - Leucovorin - Leukeran _ Leukine - Leuprolide ~ Lu , Leurocristine - Leustatin - Lipid Ara-C - Liquid Pred - Lomustine - L-PAM • L-Sarcolysin - Meticorten • Mitomycin - Mitomycin-C-Mitoxantrone-prednisol-MTC-MTX-Mustargen-Mustine-Mutamycin - Myleran - Iressa - Irinotecan • Isotretinoin - Kidrolase - Lanacort - L-Day Winter Streptokinase-LCR In several embodiments, interferon (IFN) can be used in combination with a compound of the invention. Suitable interferons include: interferon a-2a' interferon a_2b, pegylated interferon a including interferon a-2a and interferon a-2b, interferon beta, interferon beta, interferon tau, interferon ω, INFERGEN (interferon acon_i) (made by InterMune), OMNIFERON (natural 88 200902034 interferon) (made by Virage), Ou Buffon (ALBUFERON) (manufactured by Human Genome Sciences), REBIF (interferon beta-la) (made by Ares-Serono), omega interferon (Manufactured by Bi〇Medicine 5), oral interferon alpha (manufactured by Amarillo Biosciences), and interferon gamma, interferon tau, and/or interferon gamma-1β ( Manufactured by the company.) In one embodiment, the triclinide prodrug, TCN, TCN-P, and related compounds as disclosed herein can be used in combination with an additional chemotherapeutic agent such as the additional chemotherapeutic agent described herein or in Table 3 or Used interchangeably to treat drug-resistant cancers such as multi-drug resistant cancers. Drug-resistant cancers include colorectal cancer, bone cancer, kidney cancer, adrenal cancer, pancreatic cancer, liver cancer, and/or any other cancer known to the artisan or as disclosed herein. In one embodiment, the additional chemotherapeutic agent is a sputum-glycoprotein inhibitor. In several non-limiting example 15, the 'p-glycoprotein inhibitor can be selected from the group consisting of verapamil, cyclosporin (such as cyclosporine), tamoxine, Calmodulin antagonists, dexverapamil, dexniguldipine, valspodar (PSC 833), biricodar (νχ_71〇), Tower 20 tariquidar (XR9576), Zuo Suqida (Z0SUqUidar) (LY335979), aniiqUidar (R1〇1933) and / or ONT-093 〇 9. Pharmaceutical composition

Suitable for administration of the tromethamine prodrug, TCN, TCN-P 89 200902034 and related compounds as provided herein, and the acceptable carrier, including those skilled in the art: &amp; Any such carrier used for the specific administration of the drug *^^. These compounds may be formulated as a single pharmaceutically active ingredient or may be formulated in combination with other active ingredients. 5 10 15 3 The composition of the tromethamine prodrug, TCN, TCN-P and related compounds disclosed herein is suitable for oral, rectal, nasal, trans-cheek and sublingual, and Vaginal or parenteral (including subcutaneous, intramuscular subcutaneous, intradermal, intraocular, intrabronchial, intracisternal, intraabdominal, and epidural) administration. (4) The composition is administered intravenously. The compositions may conveniently be packaged in unit dosage form or may be prepared by conventional pharmaceutical techniques. Such techniques include the step of combining one or more of the present invention groups with one or more pharmaceutical carriers or excipients. The tromethamine prodrug, TCN, TCN-oxime and related compounds can be formulated into appropriate pharmaceutical preparations such as solutions, suspensions, troches, dispersions, pills, capsules, powders, sustained release formulations, or elixirs. The agent is administered orally; or it may be administered as a sterile solution or suspension for parenteral administration; and a transdermal patch preparation and a dry powder inhaler. In one embodiment, the aforementioned triclinide compound can be formulated into a pharmaceutical composition using techniques and procedures well known in the art (for example, see Ansel, Introduction to Pharmaceutical Formulations, 4th Ed., 1985 20, p. 126). In the composition, an effective concentration of one or more of the triclinide prodrug, TCN, TCN-oxime, and related compounds, or a pharmaceutically acceptable derivative thereof, may be combined with one or more suitable pharmaceutical carriers. The compound of the present invention can be derivatized into corresponding salts, esters, enol ethers or enol esters 90 200902034, acetals, ketals, protopiums + s, day, half, Secret, semi-metastatic, acid, alkali, solvate, human hydrate, or prodrug. The concentration of the compound in the composition can be administered to the therapeutically, prophylactically or ameliorating amount of the one or more symptoms of the target disease or condition. In one embodiment, the composition is formulated for administration in a single dose. In order to formulate the composition, the weight component of the compound is mixed with the selected system by other means, so that the condition to be treated is relieved, _, or one or more symptoms are ameliorated. 10 15 20 Suitable oral pharmaceutical compositions may be in separate units such as, but not limited to, a key sword, an elliptical bond, a pill, or a sugar coating, a capsule, or a sachet, and each containing a predetermined amount of one or more of the compositions; A powder or granule, a solution or a liquid solution in a liquid or non-aqueous liquid; or an oil-in-water emulsion or a water-in-oil emulsion or a bolus preparation. The liquid pharmaceutically acceptable composition can be prepared, for example, by dissolving, dispersing or otherwise mixing the triclinide compound and the optional pharmaceutical adjuvant into a carrier such as water, saline, aqueous glucose, glycerol, glycine. Alcohol, = alcohol, etc., II dissolve the solution float. If desired, the pharmaceutical composition to be administered may also contain small amounts of non-toxic auxiliary substances such as wetting agents, emulsifying agents 7, solubilizing agents, pH buffering agents, reducing agents, flavoring agents, etc., such as acetate, lemon, sodium, and rings. A dextrin derivative, sorbitan monolaurate, triethanolamine = sodium, triethanolamine oleic acid vinegar, and other such adjuvants. Such dosage forms are known to those skilled in the art or are readily known, for example, to Remington Pharmaceutical Sciences, Merck Publishing Company, Easton, Pennsylvania, 15th Edition, 1975. Compositions of the invention suitable for topical administration to the mouth include, for example, buccal ingots, 91 200902034 which contains ingredients for the flavoring base which are typically sucrose and acacia or tragacanth; softening agents containing one or more The compositions of the invention are in inert bases such as gelatin and glycerin or sucrose and acacia gum; and mouthwashes containing one or more of the compositions of the invention administered in a suitable liquid carrier. 5 tablets, pills, capsules, throat tablets and the like may contain one or more of the following ingredients or compounds having similar properties: binder; lubricant; diluent; slip agent; disintegrating agent; coloring agent; sweetener; Wetting agent; emetic coating; and film coating. Examples of the binder include microcrystalline cellulose, tragacanth, glucose solution, acacia, gelatin solution, molasses, polyvinyl 10-pyrrolidone, povidone, cross-linked Pvillon, Sucrose and starch paste. Lubricants include talc, starch, magnesium stearate or stearic acid, lycopodium, and stearic acid. The diluents include, for example, lactose, Yan sugar, starch, kaolin, salt, mannitol, and dicalcium phosphate. Sliding agents include, but are not limited to, colloidal cerium oxide. Disintegrating agents include crosslinked methylcellulose sodium, 15 sodium starch glycolate, alginic acid, corn starch, potato starch, bentonite sulfhydryl cellulose, agar, and carboxymethyl cellulose. The coloring agent includes, for example, any of the finely approved and approved water-soluble FD&amp;C dyes, a mixture thereof; a water-insoluble FD&amp;C dye suspended on hydrated alumina. Sweeteners include sucrose, lactose, mannitol, and artificial sweeteners such as saccharin and spray-dried. Bridge odorants include synthetic flavors extracted from plants such as fruits, and synthetic blends of compounds which produce a pleasant sensation, such as, but not limited to, menthol and methyl salicylate. Wetting agents include glyceryl monostearate, sorbitan monooleate, diethylene glycol monolaurate, and polyoxyethylidene lauryl ether. The emetic coating includes fatty acids, fats, waxes, 92 200902034 shellac, ammoniated shellac, and cellulose acetate phthalate. Film coats include hydroxyethyl cellulose, sodium carboxymethyl cellulose, polyethylene glycol 4000, and cellulose acetate phthalate. Compositions suitable for topical administration to the skin may be formulated with one or more of the compositions 5 in ointments, creams, gels, and pastes in a pharmaceutically acceptable carrier. The rectal pharmaceutical compositions can be administered in the form of a suppository containing a suitable base such as cocoa butter or a salicylate. Suitable for nasal administration, when the carrier is a solid, the composition 10 comprises a coarse powder having a particle size ranging from 20 micrometers to 500 micrometers, which is administered as a nasal olfactory powder (ie, the powder container is kept close to the nostrils) Rapid inhalation through the nasal passages). When the carrier is a liquid (e.g., a nasal spray or a nasal drop), one or more of the compositions may be mixed into an aqueous solution or an oily solution to be inhaled into the nasal passages or sprayed into the nasal passages. 15 A composition suitable for vaginal administration may be in the form of a pessary, tampons, ointment, gelatin, paste, foaming or spray formulation containing one or more of the compositions and a suitable carrier. Compositions suitable for parenteral administration include aqueous and non-aqueous sterile injectable solutions, which may contain antioxidants, buffers, bacteriostatic agents, and solutes which allow the 20-part to become isotonic with the blood of the intended recipient, and aqueous Sterile suspensions and non-aqueous sterile suspensions, including suspending and thickening agents. The composition may be in the form of a single or multiple dose containers such as sealed ampoules and sealed vials, and the composition may be stored under lyophilization (lyophilization) conditions by the addition of a sterile liquid carrier such as water for injection just prior to use. Temporary injection solutions 93 200902034 and suspensions can be prepared from the sterile powders, granules and lozenges described above. Medicinal organic or inorganic solid or liquid carrier media suitable for enteral or parenteral administration can be used to make the compositions. Gelatin, lactose, starch, magnesium stearate, talc, vegetable and animal fats and oils, gums, polyalkylene glycols, water, or other known carriers are all suitable as carrier media. The composition can be used in combination with the active ingredient in one or more pharmaceutically acceptable carrier vehicles and/or excipients. As used herein, "pharmaceutically acceptable carrier medium" includes any and all carriers, solvents, diluents, or other liquid vehicles, dispersing or suspending aids, surfactants, isotonics. Agent, thickener or emulsifier, preservative, solid binder, lubricant, adjuvant, vehicle, delivery system, disintegrating agent, absorbent, preservative, surfactant, colorant, flavor, or sweet Flavoring agents and the like are used depending on the particular dosage form that is suitable for use in the 15th. In addition, the compositions can be combined with pharmaceutically acceptable excipients and, optionally, sustained release matrices, such as biodegradable polymers, to form a therapeutic composition. "Pharmaceutically acceptable excipients" include non-toxic solid, semi-solid or liquid filters, diluents, encapsulating materials or formulation adjuvants of any type. 20 However, it is important to understand that the total dose used for each component is determined by the clinician based on the scope of the complete medical decision. The particular therapeutically effective amount for any particular host will be determined by a number of factors, including, for example, the condition being treated and the severity of the condition; the activity of the particular composition employed; the particular composition employed, the patient's age, weight, and general Health status, gender and diet; 94 200902034 Time of administration; route of administration; excretion rate of the particular compound used; time of treatment; factors such as those commonly used in combination with or in combination with the particular composition employed. For example, it is known within the skill of the industry to start administering the composition at a dose that is 5 lower than the dose required to achieve the desired therapeutic effect, and to slowly increase the dosage to achieve the desired efficacy. The composition is preferably formulated into a unit dosage form for ease of administration and uniform dosage. &quot;Unit dosage form&quot; as used herein refers to a physically separate unit suitable for use in the compositions of the host to be treated. Each dose must contain a quantity of the composition calculated to produce a desired therapeutic effect in combination with or in combination with the selected pharmaceutical carrier medium. Preferred unit dosage formulations are unit dosage formulations containing the daily or permissible dosage unit, per unit dosage, or suitable component thereof. For example, about 1-5 mg of the compound disclosed herein can reduce the tumor volume of the mouse. The dosage will be determined by host factors such as body weight, age, surface area, metabolism, tissue distribution, rate of absorption, and rate of excretion. In one embodiment, from about 0.5 grams to about 7 grams of the compound disclosed herein can be administered to a human. Optionally, from about 1 gram to about 4 grams of compound per day can be administered to humans. In several embodiments, about 0.001 mg to 5 mg of triclinbine compound per day can be administered to humans. The effective dose for treatment will be determined based on a number of factors as described above. In addition, within the skill of the art, it is possible to start administering the composition at a relatively low dose and to increase the dosage to achieve the desired effect. A composition comprising a tromethamine prodrug, TCN, TCN-P, and 95 200902034, as disclosed herein, can be used for sustained release of a matrix which can be resolved by acid hydrolysis or by dissolution. Made of decomposed material, subject to _ usually polymer. Once the substrate is embedded in the human body (4), the matrix phase &amp; Saki and body fluids. The sustained release matrix is, for example, selected from the group consisting of: biomaterials such as micro-records, (iv) cross (iv) (10) called polyacetate acetate, 夂t °), polylacide-co-glycolic acid (lactic acid and glycolic acid) Co-white, phthalic anhydride poly (original 酉夂) 5 categories, peptides, hyaluronic acid, collagen egg yolk / &amp; soft osin, New Zealand, fatty acids, coffee 1 class, polysaccharides, ^ polyamino acids 'Amino acids such as phenylalanine, sulphate, sulphate, 12 nuclear acid, polyvinyl propylene, polyvinyl carbaryl ester. The preferred biodegradable matrix is polylactide, poly Ethylene or Polypropylene turtle _ Copolymer · Ethyl alcohol (the base of lactic acid and glycolic acid - 15 materials to receive the West Li) negative precursor drugs, TCN, TCN_P and related compounds can also be used for the month: ° b (four) k know 'Microlipids (4) are often derived _ = their mussels. The vesicles are formed by single or multi-layer hydrated liquid crystals. The nucleus can be used to form any lipid-free granules. Receptive and metabolizable moon essence. In addition to the present invention, the seed or the scorpion stalk may contain stabilizers and deposits. , excipients, etc. Examples of 14 are natural and synthetic phospholipids and phospholipid choline (impressed phosphorus) = lipid granule formation method is well known in the art. Breathing head _ can be formulated into an aerosol for, for example, by inhalation The method of administration can be carried out in the form of an aerosol or a spray solution or in the form of a fine powder for human use, and can be used alone or in combination with an inert carrier such as lactose to use 96 200902034. In one embodiment, the formulation particles have a diameter of less than 50 microns, and in one embodiment, the diameter is less than 1 micron. The inclusion of the triclinide prodrug, TCN, TCN_p, and related compound compositions disclosed herein. It can be used in combination with other compositions and/or procedures for treating the aforementioned conditions. For example, tumors are conventionally treated with a combination of one or more of the compositions of the invention using a 'radiotherapy or chemotherapy combination, followed by one or more of the inventions. The composition can be administered to the patient to prolong the dormancy of microscopic tumor metastasis and to stabilize, inhibit or reduce the growth of any residual primary tumor. The pharmaceutical compositions of the present invention can be formulated according to known methods for pharmaceutically useful compositions. The formulations are described in a variety of sources well known and readily available to those skilled in the art. For example, Remington Pharmaceutical Sciences (Martin EW [1995], Etchton, Pa., 19th edition of Merck Publishing Co., Ltd. describes formulations that can be used in connection with the present invention. Formulations suitable for administration include, for example, aqueous and non-15 aqueous sterile injectable solutions, which may contain antioxidants, buffers, The bacteriostatic agent, and the solute which makes the formulation is isotonic with the blood of the intended recipient, and the aqueous sterile suspension agent and the non-aqueous sterile suspension agent, which comprises a suspending agent and a thickening agent. The formulation may be in a single dose or multiple doses. The container is in the form of, for example, a sealed ampoule and a sealed vial, and the composition can be stored under lyophilization (lyophilization) conditions, requiring only 2 liters of sterile liquid carrier such as water for injection to be added just prior to use. Temporary injection solutions and suspensions can be prepared from sterile powders, powders, lozenges and the like. It will be appreciated that in addition to the ingredients specifically described above, the formulations of the present invention may include other chemical agents known to those skilled in the art of this type of formulation. The method of the present invention is used, for example, to inhibit the growth of cancerous cells, and the method of the present invention is preferably combined with at least one other method of treatment including, but not limited to, chemotherapy, radiation therapy, selective inhibition of Ras oncogene signaling, or Any other treatment known to those skilled in the art of cancer treatment and disposal, such as administration of an anticancer agent.

5 It is also possible to carry out salt formation and to give triclinide prodrug, TCN, TCN-P and related compounds. Examples of pharmaceutically acceptable salts are organic acid addition salts with acids which form physiologically acceptable anions, such as tosylate, methanesulfonate, acetate, citrate, C. Diacid salts, tartrates, succinates, benzoates, ascorbates, (X-ketopentane 10 salts, and a-glyceryl phosphates. Suitable mineral acid salts, including sulfates, may also be formed. Nitrate, bicarbonate, and carbonate. Pharmaceutically acceptable salts can also be obtained using standard procedures well known in the art, such as the reaction of a sufficiently basic compound such as an amine with an appropriate acid which forms a physiologically acceptable anion. It is also possible to produce an alkali metal (for example, 15 sodium, potassium or chain) salt or a soil test metal (for example, mom) salt. The triclinide prodrug, TCN, TCN-P and related compounds of the present invention can be formulated into a pharmaceutical. The composition, and in a plurality of selected administration forms, that is, oral or parenteral, is administered to an individual such as a patient or a sick animal by an intravenous route, a muscle route, a local route or a subcutaneous route. 20 Thus, the Quxi of the present invention Pre-prodrug, TCN, TCN-P and related compounds may be administered in combination with a pharmaceutically acceptable vehicle (ie, a carrier) such as an inert diluent or an assimilable edible carrier. It may be encapsulated in a hard or soft shell. The gelatin capsule can be compressed into a tablet or can be directly mixed with the patient's food. For oral administration, the compound is combined with one or more of the ingredients 98 200902034, and can be used as a feeding lozenge, buccal ingot, and throat. , capsules, elixirs, suspensions, syrups, pastes, etc. These compositions and preparations must contain at least 0.1% active agent. The percentage of the composition and the preparation may of course be changed, conveniently given A range of from about 2% to about 60% by weight of the unit dosage form. The active compound content of such a therapeutically useful composition is an effective dosage level obtainable. Tablets, guillotine, pills, capsules, and the like are also included. The following ingredients: binders such as tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; disintegrating agents such as corn starch, potato starch, alginic acid 10, etc.; lubricants Such as magnesium stearate; and sweeteners such as sucrose, fructose, lactose or aspartame; and flavoring agents such as menthol, wintergreen oil or cherry flavor. When the unit dosage form is a capsule, in addition to the materials of the aforementioned categories In addition, the capsules contain a liquid carrier such as vegetable oil or polyethylene glycol. There may be many other forms of coating, or otherwise modify the shape of the solid unit dosage form. For example, lozenges, pills or capsules It may be coated with a gelatin, wax, shellac or sugar, etc. The syrup or elixir may contain a compound of the invention, sucrose or fructose as a sweetener; decyl paraben and propylparaben as a preservative; Dyes and flavors such as cherry or orange flavor. Of course any material used in the preparation of any unit dosage form must be pharmaceutically acceptable 20 and substantially non-toxic in its amount used. Furthermore, the compounds of the invention may be incorporated into sustained release. Formulation and sustained release device. The active agent (e.g., API-2, a prodrug or a pharmaceutically acceptable salt thereof) can also be administered intravenously or intraperitoneally by infusion or by injection. A solution of the active agent or a salt thereof can be prepared by mixing a non-toxic surfactant in water as needed. Fraction 99 200902034 Dispersing agents can also be prepared from glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof, and in oils. Under ordinary conditions of storage and use, such preparations contain a preservative to prevent the growth of microorganisms. The pharmaceutical dosage form suitable for injection or infusion comprises a sterile aqueous solution or an aqueous dispersion or a sterile powder containing the active ingredient, which is suitable for the preparation of a sterile injectable solution or an infusion solution or dispersion, if necessary. Encapsulated in microlipids. In summary, 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, polyol (for example, glycerol 10, propylene glycol, liquid polyethylene glycol, etc.), vegetable oils, non-toxic glycerides, and suitable mixtures thereof. . Suitable fluid properties can be maintained, for example, via the formation of vesicles, by the maintenance of the desired particle size in the case of dispersions, or by the use of surfactants. Prevention of the action of microorganisms can be obtained by various antibacterial agents and antifungal agents, for example, p-hydroxybenzoic acid esters, chlorobutanol, phenol, sorbic acid, and thimerosal. In many cases, it may be preferred to include an isotonic agent such as a saccharide, a buffer or a sodium chloride = composition for injection by prolonged absorption by the use of a delayed absorbent such as aluminum stearate and gelatin in the composition. Sterile injectable solutions are prepared by incorporating the compound of the present invention in a suitable solvent, if necessary, in the presence of the above ingredients, followed by filtration of the bacteria. In the case of a sterile powder for the preparation of a sterile injectable solution, the preparation method is vacuum drying and freeze-drying techniques to obtain the active ingredient plus any additional ingredients which are desired to be present in the sterile filtered solution. For topical administration, the compounds of the invention may be administered in neat form, also as 100 200902034 in liquid form. However, it is generally desired that the compound of the present invention can be applied to the skin in combination with the skin (4). The elixirs can be solid or liquid and applied to the skin in the form of a composition or formulation. Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, cerium oxide, oxidized imitation, and the like. Useful liquid carriers include water, lysine or gan or L-glyconate, and the compounds of the invention may be dissolved or dispersed in an effective amount by means of a surfactant-free I if desired. Adjuvants such as perfumes and additional antimicrobial agents can be added to achieve optimal properties for a given use. The resulting liquid composition can be applied by an absorbent pad for impregnating the 10-end tape and other dressings, or sprayed onto the affected area using a pump-type sprayer or aerosol sprayer. Thickeners such as synthetic polymers, fatty acids, fatty acid hydrochloric acid, and fatty acid esters, fatty alcohols, modified celluloses, or modified mineral materials can also be used in liquid carriers to form expandable pastes, gels. The agent, ointment, 15 soap, etc. are for direct application to the skin of the user. Examples of useful dermal compositions that can be used to deliver a compound of the invention to the skin are disclosed in Jacquet et al. (U.S. Patent No. 4,608,392), to Geria (U.S. Patent No. 4,992,478), to Smith et al. (U.S. Patent No. 4,559,157). And Woltzman (U.S. Patent 4,820,508). The useful dose of the pharmaceutical composition of the present invention can be determined by comparing its activity in vitro and in vivo activity in an animal research model. Methods for extrapolation to humans in effective doses in mice and other animals are known in the art; for example, reference is made to U.S. Patent No. 4,938,949, to a non-limiting example, in a body composition such as lotion 101 200902034 The concentration of the agent may be about 0·1 -25 wt.-Q/. Or by about 0.5-10 wt_-%. In one embodiment, the concentration in the semi-solid composition or solid composition such as gel or powder is from about 0.1 to about 5 wt.-°/〇, preferably from about 0.5 to 2.5 wt.-%. In one embodiment, a single dose of &apos;injected, infused or ingested is typically 5-1500 54: grams&apos; and may be administered once daily to achieve a concentration of about 0.1-50 milligrams per kilogram in an adult. The non-limiting dose of the present invention is 7.5 mg to 45 mg per ounce of oral administration, and the individual body weight can be appropriately adjusted. Thus, the invention includes a pharmaceutical composition comprising a combination of a triclinide prodrug, a TCN, a TCN-P, and a related compound, or a pharmaceutically acceptable salt thereof, as disclosed herein, and a pharmaceutically acceptable carrier. Pharmaceutical compositions suitable for oral, topical or parenteral administration include a quantitative triclinide prodrug, TCN, TCN-P and related compounds, or a pharmaceutically acceptable salt thereof, which composition preferably comprises the present invention. Example. In the context of the present invention, the dosage administered to an individual, particularly to the human body, is sufficient for a reasonable time frame to affect the therapeutic response of 15 patients. Those skilled in the art will understand that the dosage will be determined based on a number of factors, including the animal's condition, the animal's weight and the severity of the cancer, and the stage of the cancer. The appropriate dose is to obtain the concentration of the active agent known to exert the desired response in the tumor tissue. The preferred dose is that the cockroach has the greatest inhibitory effect on the growth of cancer cells, and there is no uncontrollable side effect. The administration of triclinide prodrug, TCN, TCN-P and related compounds (except for the sputum; the upper salt) can be administered continuously or at separate intervals, as determined by skilled persons in the art world. . Mammals 102 may benefit from the disclosed methods of inhibiting the growth of cancer cells 200902034 - including but not limited to primates such as baboons, chimpanzees, orangutans, humans, monkeys; cloisters such as pets such as dogs, cats, guinea pigs , hamsters, Vietnamese pot-bellied pigs, and ferrets; farm livestock such as cattle, buffalo, bison, horses, donkeys, pigs, sheep, and I are Jiushanping; typical rare animals such as bears, 5 lions, commonly found in zoos, Tiger, heat a, elephant, seahorse, rhino, giraffe, antelope, sloth, gazelle, zebra, earth &amp; „ ^ Ling, groundhog, koala, kangaroo, possum, &gt; Tian j 1 dog, seal, sea lion, walrus, otter, bottlenose dolphins, dolphins contend. ", this 丨... two people" and "individual" are used interchangeably here, with the intention of including human milk animals and non-human Mammalian species. For the same reason, the test tube test can be used for cells of this mammalian species in July and July. Patients to be treated using the methods of the present invention can be identified using standard techniques known to those skilled in the medical arts. The following examples are provided by way of illustration and not limitation. Οο. SM. 15 10丄 Example 1: Tube inside the sieve; ^ Materials laj|_ and NCI diversity sets | : All cell lines are available from ATCC or as previously described (Cheng JQ et al., Oncogene, 14: 2793) -2801, 1997; West KA et al., Drug Resistance Today, 5: 234-248, 2002; 20 Satyamoorthy K. et al., Cancer Research, 61: 7318-7324, 2001). The NCI structure diversity set is a library of 1,992 compounds selected from a pool of approximately 140,000 compound NCI drugs. Further information on the selection, structure and activity of these diversity collection compounds can be found in the NCI Development Treatment Program website. 103 200902034 Screening for cell growth inhibition by Akt translocation: NIH3T3 cells transfected with AKT2 or NIH3T3 control cells infected with LXSN vector (Cheng JQ et al., Oncogene, 14: 2793-28 (H, 1997) was inoculated into a 96-well tissue culture well plate. After treatment with 5 μΜΝα diversity collection compound 5, the cell growth line was detected using Cell Tier 96-solution solution cell proliferation kit group (Promega). In NIH3T3 cells transfected with AKT2-transformed NIH3T3 cells but not infected with LXSN, compounds that inhibit growth were considered candidates for Akt inhibitors for further analysis. 10 Intracellular protein kinases, cell survival, and apoptosis Test scores; ^ : In vitro kinases were performed as described above (for example, refer to Jiang K. et al., Mol Cell Biol, 20: 139-148, 2000). Cell survival was assayed by MTS (Pumiga) assay. It was detected by annexin V as described above (Jiang K., Coppola et al., Mol Cell 15 Biol, 20: 139-148, 2000). Recombinant Akt and PDK1 were purchased from the Upper Biotechnology Company ( Upst Ate Biotechnology Inc) 〇 result

The small molecule of Akt signaling pathway inhibits 刽API-2 as a result of the detection of Akt changes in human cancer, destroying the Akt pathway, inducing apoptosis and inhibiting tumor growth (Jetzt A• et al., Cancer Research, 63: 697-706, 2003). Thus 'Akt is seen as an attractive molecular target for the development of novel cancer therapeutics. In order to identify small molecule inhibitors of Akt, 1,992 compound chemical libraries (NCI diversity sets) from NCI were evaluated for inhibitors of NIH3T3 fine 104 200902034 cells transfected with AKT2-transformed but not transfected with blank vector LXSN. . Repeat experiments showed that 32 compounds could only inhibit growth by AKT2-transformed cells. One of the most potent compounds, API-2 (NCI identifier: NSC 154020), inhibits cell growth at a concentration of 50 nM. Fig. 1A shows that API-2 is also referred to as the chemical structural formula of Tricinem (Schweinsberg P.D. 5 et al., Biochem Pharmacol, 30: 2521-2526, 1981). The fact that API-2 selectively inhibits AKT-2 transfected cells over untransformed parental cells reminds the inventors whether API-2 is an AKT2 kinase inhibitor. To achieve this, AKT2 was immunoprecipitated with anti-AKT2 antibody from NIH3T3 cells transfected with AKT-2 after treatment with API-2. AKT2 immunoprecipitation was performed using an anti-phospho-Akt antibody for immunological dot analysis. As shown in Figure 1B, API-2 significantly inhibits AKT2 phosphorylation in serine-309 and serine-474 (both required for complete activation of AKT2) (Datta SR et al., Gene Dev, 13: 2905- 2927, 1999). Since the isoforms of the three Akt share a highly homologous and similar structural formula, the effect of API-2 on its kinase activity was evaluated. 15 HEK293 cells were treated with HA-Aktl, -AKT2 and -AKT3 prior to stimulation with EGF (50 Ng/ml), serum was depleted overnight and treated with API-2 for 60 minutes. Three experiments were repeated showing that API-2 inhibits EGF-induced kinase activity and phosphorylation of Aktl, AKT2 and AKT3 (Fig. 1C). However, the kinase activity of the recombinant strain consisting of active AKT2 (Myr-AKT2) is inhibited by API-2 in the in vitro kinase reaction (Fig. 1D), suggesting that API-2 does not directly inhibit Akt in the test tube, API-2 Neither as an ATP competitor nor as an enzyme matrix competitor that binds to the active site of Akt. API-2 does not inhibit the known activation of Akt upstream: it is clearly documented in the literature that Akt is activated by extracellular stimuli and intracellular signaling molecules such as active Ras and Src through the 105 200902034 PI3K-dependent manner. Thus, targeting upstream molecules of Akt will result in inhibition of Akt by API-2. Since PI3K and PDK1 are direct upstream regulators of Akt (Datta S. R. et al., Gene Dev, 13: 2905-2927, 1999), it was examined whether API-2 inhibits PI3K and/or PDK1. HEK293 cells were deficient in serum, and then treated with ΑΠ-2 or PI3K inhibitor, Watmanin for EGF stimulation for 30 minutes. PI3K was immunoprecipitated with an anti-ρ11〇α antibody. The immunoprecipitate was assayed by in vitro assay using PI-4-P as the enzyme substrate. As shown in Figure 2, EGF-induced Κ3Κ activity was inhibited by Watmanin but not by ΑΡΙ-2. To assess the effect of ΑΠ-2 on PDK1, assays for recombinant PDK1 to promote the phosphorylation of threonine-309 phosphorylation of ΑΚΤ2 polypeptide were used in the presence of phospholipid-containing myothiol-containing lipid vesicles. As shown in Figure 2, the assay was strongly inhibited by the control PDK1 inhibitor, sturosporine (IC50 = 5 ηΜ). Contrary to the highest test concentration (5·ΐμΜ), ΑΡΙ-2 showed only 21% inhibition of the assay. To further evaluate the effect of ΑΡΙ-2 on PDK1 activation, the degree of autophosphorylation of PDK1 in serine _241 was examined after 15 ΗΕΚ293 cells were treated with ΑΡΙ-2. Serine _241 was autophosphorylated and active. A residue of critical importance (Datta SR et al., Gene Dev, 13: 2905-2927, 1999). Repeated three experiments showed that the degree of phosphorylation of pDK1 was not inhibited by API-2 (Fig. 2B). However, the ρΙ3κ inhibitor Watmanin inhibits 20 EGF-stimulated PDK1 (Fig. 2). △ΡΙ-2对逸过Ρ|ςς^ΚΑ, SGK:, STAt, JNK, Ρ38

After IERK sends out the route Selective: Asahi belongs to the AGC (PKA/PKG/PKC) kinase family, which also includes ρκΑ, PKC, serum-induced lining and glucocorticactivator (SGK), and sucrose 200902034 S6 kinase p7〇, mitogen- and stress-activated protein stimulating plum and PKC-related stimuli. In the AGC kinase family, the structural structures of PKA, PKC and SGK are closer to the protein structure of Akt kinase than other members. Therefore, the second test API_2 affects the catalytic activity of the three kinases. HEK293 cells were infected with HA-tagged PKA, PKCa or SGK. Intracellular kinase assay analysis and immunoblot analysis showed that the kinase activities of pKA and PKCa were inhibited by a PKC inhibitor, namely PKAI and Ro 31 '8220, respectively; API-2 had no effect on its activity (2C and 2E). Figure). In addition, serum-inducible SGK kinase activity was attenuated by Wattmannin but 10 was not attenuated by API-2 (Fig. 2D). In addition, it was determined whether API-2 had an effect on the survival pathway of other oncogenes. Western blot analysis using commercially available anti-phospho antibodies showed that the degree of phosphorylation of Stat3, JNK, p38 and Erkl/2 was not affected by apj_2 treatment (Fig. 2F). These data indicate that API_2 specifically inhibits the Akt signaling pathway. 15 Old 1.4.kt overexpressed/activated human cancer cells API API-2 stops cell growth and induces apoptosis ● API-2 selectively inhibits Akt pathway, suggesting a preference for Akt disorder These tumor cells that express/activate inhibit proliferation and/or induce apoptosis. API-2 is used to treat overexpression of 20 AKT2 (via OVCAR3, OVCAR8, PANC1, and AKT2-transformed NIH3T3) or PTEN gene mutations due to Akt overexpression or PTEN mutations leading to activation of Akt in human malignancies ( PC-3, LNCaP, MDA-MB-468) prime cells that express active Akt, as well as cells that do not express (OVCAR5, DU-145, T47D, C0L0357, and LXSN-NIH3T3), and are activated by IGF-1 activation. Akt or melanoma cells that do not respond to growth stimulation by IGF-1 107 200902034 (Satyamoorthy et al., Cancer Research, 61: 7318-7324, 2001). Immunoblot analysis showed that the degree of phosphorylation of Akt was inhibited by API-2 only in cells that exhibited elevated Akt or responded to IGF-1 stimulation (Fig. 3A). Thus, API-2 inhibits cell growth in Akt 5 overexpressing/activating cells to a much higher degree than cells containing low levels of Akt. As shown in Figure 3B, in the Akt overexpression/activated cell line, LNCaP, PC-3, OVCAR3, OVCAR8, PANC1, MDA-MB-468, and WM35, API-2 treatment inhibited cell proliferation by approximately 50-60. %; cells in DU145, OVCAR5, C0L0357, T47D, and WM852 10 inhibited cell proliferation by only about 10-20%, and these cells contained low concentrations of Akt, or did not respond to growth stimulation by IGF-1. In addition, API-2 induced cell apoptosis by 8-fold (OVCAR3), 6-fold (OVCAR8), 6-fold (PANC1), and 3-fold (AKT2-NIH3T3). In OVCAR5, C0L0357, and LXSN-NIH3T3 cells, no significant differences in apoptosis were observed between API-2 treatment and vehicle (DMSO) treatment. Thus, in the performance of disordered Akt, API-2 prefers to inhibit cell growth and induce apoptosis in cells exhibiting disordered Akt. API-2 inhibits downstream targeting of Akt: Akt has been shown to exert its cellular effects through phosphorylation of various proteins (Datta S. R. et al., Gene Dev, 13: 2905-2927, 1999). More than 20 proteins have been identified as Akt enzyme substrates, 20 including inserter protein family members (FKHR, AFX, and FKHRL1), nodule 〇1^ soil 11)/丁8匚2, 卩7〇, 〇81( : -30, 卩21'^1/(^1, mouth 27, 1, MDM2, Bad, ASK1, ΙΚΚα, etc.. Secondly, whether API-2 inhibits Akt downstream targets. Because of anti-phospho- nodulin, _Bad, The -AFX, and -GSK-3P antibodies are commercially available, and therefore the effect of API-2 on the induction of phospho-108 200902034 by Akt was determined. After treatment with ΑΡΙ-2 (ΙμΜ), OVCAR3 cells were lysed and individual anti-antigens were used. Phosphoric antibodies were used for immunoblotting analysis. Figure 4 shows that ΑΡΙ-2 significantly inhibits the degree of phosphorylation of nodulin, resulting in the stability and upregulation of nodulin (Dan, H.C_ et al, J Biol Chem, 277: 35364- 35370, 5 2002) 〇

The degree of phosphorylation of Bad, GSK-3p, and AFX was partially attenuated by ΑΡΙ-2. These data suggest that ΑΠ-2 induces cell death and cell growth arrest by inhibiting phosphorylation of its downstream targets. The possible reason for API-2 to inhibit Akt downstream targets in varying degrees is that the phosphorylation sites of these targets are also regulated by other kinases 10. For example, Bad serine-136 is also used in addition to Akt phosphorylation. Phosphorylation (Schurmann, A. et al., Mol Cell Biol, 20: 453-461, 2000). 10·2·Example 2: Anti-tumor activity in sputum xenografts in nude mice 15 Tumor cells were harvested, suspended in PBS, and injected subcutaneously into the right side of 8-week-old female nude mice as previously reported. With the left flank (2χ1〇6 cells/flank) (Sun J. ' Blaskovic et al., Cancer Research, 59: 4919-4926, 1999). When the tumor size reached approximately 100-150 mm 3 , the animals were randomly assigned and injected intraperitoneally with 0.2 ml of drug vehicle. Control animals received 20 DMSO (20%) vehicle, while treatment animals were injected with API-2 (1 mg/kg/day) in 20% DMSO. API-2 shows the growth of AKT 1 and AKT2 in human ovarian cancer and pancreatic cancer in the presence of Akt extreme mouse overexpression/activation and/or amplification (〇^11§ 1. (5. and Sichuan (: 08丨3 8.\^, the tumor occurred in the eight feet 7' letter 109 200902034 transduction path, edited in Schwab D, cancer encyclopedia reference, Berlin, Heidelberg and New York' Springer; 2001 The 35-37 hAkt pathway is inhibited by inhibitors such as pI3K, HSP70, Src, and farnesyltransferase, leading to cell growth arrest and induction of apoptosis (Solit D. B. et al., Cancer Research, 63: 5). 2139-2144, 2003; Xu, W. et al., Cancer Research, 63: 7777_7784, 2003). Recent studies have shown that tumor growth in xenografts with elevated Akt is also significantly inhibited by intratumoral injection of dominant negative Akt adenovirus (Jetzt A. et al., Cancer Research '63: 697-706, 2003). Inhibition of Akt by API-2 and induction of apoptosis and cell growth arrest in cancer cells with elevated Akt levels 10 (Fig. 3) Therefore, tumor growth with elevated Akt content should compare tumor growth of tumors with lower Akt in nude mice, and be more sensitive to API-2. In order to achieve this, subcutaneous injection of Akt overexpressing cells (〇vcAR3, OVCAR8 and PANC-1) was implanted subcutaneously into the right flank, while those expressing low Akt (0VCAR5 and COL03 57) were implanted. The left flank of the mouse. When the tumor 15 reached an average size of about 100-150 mm 3 , the animals were randomized and treated with vehicle or API-2 (1 mg/kg/day) by intraperitoneal injection. As shown in Fig. 4B, the vehicle-treated 〇vcAR5 and C0L0357 tumors grew to about 800-1000 mm 3 on the 49th day after tumor implantation. The OVCAR3, OVCAR8, and PANC-1 tumors treated with the vehicle control group, It grew to about 700-900 mm 3 on the day of implantation, and the API-2 inhibited the growth of OVCAR3, OVCAR8 and PANC-1 tumors by 90%, 88% and 80%, respectively. Conversely, 'ΑΠ-2 on nude mice Cell growth of OVCAR5 and C0L0357 was minimally affected (Fig. 4B-4D and data not shown). At 1 mg/kg/day, API-2 had no effect on blood glucose concentration, body weight, activity and food intake in mice. 110 200902034 响. In the treated tumor samples, Akt activity is affected by API-2, but total Akt contains The amount is constant (Fig. 4E). Together, these results indicate that API-2 selectively inhibits the growth of tumors with elevated Akt concentrations. 1〇·3. f Example 3: TCNIL inhibits wild-type Akt stimulating activity 5 API-2 (TCN) directly inhibits wild-type Akt kinase activity induced by PDK1 in vitro (Fig. 1). These results confirm that API_2 is a direct Akt inhibitor. The potential mechanism may be API-2 binding to the PH functional site of Akt and/or threonine-308. Intracellular kinase assays were performed in a kinase buffer containing phospholipid-based inositol-3,4,5-P3 (PIP3), API-2, and tissue gland H2B as an enzyme substrate using PDK1 and Akt 10 recombinant strains. After 30 minutes of incubation, the reaction was separated by SDS-PAGE and exposed to the membrane. 10-4. Example 4: TCN has the effect of extracting TCN (API-2) in cancer-resistant face with West Platinum, paclitaxel, and Tamoxifen resistance A270CP, C-13, OVCAR433, and MCF7 /TAM fine 15 intracellular test. In these cells, API-2 overcomes the resistance of cetamine, paclitaxel, and tamoxifene. 10.5. »^15 : Cell viability assay: Cytotoxicity was measured in A549 and Colo357 using 0.1 μΜ to iOO μΜ TCN, TCNP, or TCN treated with a Gengyl group at position 6 for 48 20 hours. After the drug was cultured, the cells were harvested by centrifugation of the protease. The number was counted using a Coulter electronic particle counter, and about 100 viable cells per well were seeded in a 6-well culture dish. After 〇曰, the cells were fixed with 3:1 methanol:acetic acid, and the cell viability was determined by 4% crystal violet staining and counting the number of communities. Cell viability was calculated as the tillering efficiency of various cell ill 200902034 types and drug concentrations as untreated control cells. Stability study: Stability was assessed at 37 ° C in 100 mM phosphate buffer, pH 6 5 and rat liver homogenate. One part of the buffer stability was taken at 〇, 丨, 25 and 3 hours, mixed with cold 10% TCA and the prodrug and parent compound were analyzed by LC/MS/MS as described. For liver homogenate, samples were taken at 5, 10, 20, 30, and 90 minutes, mixed with cold tca, and then centrifuged for 10 minutes to remove precipitated liver homogenate protein. Samples were analyzed by LC/MS/MS as described. 10 Intestinal absorption - duodenal administration: male white variants of the history of the Boer Lali rat 9-10 weeks old body weight 250-350 grams fasting 18 hours but free water. Rats were anesthetized with 2-5% isoflurane (iS0flU0rane). In order to periodically draw a blood sample, the root cannula is placed in the jugular vein. The abdominal cavity was opened by a 4-5 cm midline, and the duodenum was placed. 5 ml of a 6 mg/ml drug solution (or suspension) was directly injected into the duodenum, and the intestines were placed back into the abdominal cavity, and the incision was covered with gauze. Plasma samples (_05 ml) were withdrawn over a period of 4 hours while using the LC/MS/MS method to quantify the systemic plasma concentrations of the injected prodrugs and/or parental compounds. Sample preparation: The samples were subjected to solid phase extraction using Waters CPE SPE 20 cassette (MCX). The cassette was conditioned with 1 ml of methanol and 1 ml of water. The 25 liter sample was treated with an equal amount of 2% linonic acid and loaded onto the SPE column. The column was washed with 1 ml of a 1% TFA methanol solution and 1 ml of methanol, and then eluted with 1 ml of 2% ammonium hydroxide in methanol. The extract was evaporated to dryness under a stream of nitrogen in a Turbovap solution evaporator (10) and re-modulated on a 25 ml 112 200902034 HPLC phase. The material was transferred to a micro-plug and analyzed on a Micromass Quattro II LC/MS/MS. Analysis by LC/MS/MS: Plasma samples and stability samples were analyzed by LC/MS/MS. For HPLC, 10 microliters of the entire sample was separated at a flow rate of 〇 2 &quot; 5 ml/min over a C18, 2_2 mm χίο cm column (Higgins Analytical) over a 3 minute flow time. The phase system for separation is composed of the following phases: A) 70% 0.1% citric acid in water to B) 30% acetonitrile. The MS/MS detector operates in an MRM positive acquisition mode with a cone voltage of 30 volts, an impact energy of 5 volts, and an impinging pressure of 1 χ 1 〇 3 mbar. 10 Synthesis and synthesis of tricineribine prodrugs: Two kinds of tromethamine-based prodrugs based on the 5' hydroxyl group of tromethamine And b) 5' 〇-hydrazinyl phosphatidyl citrate, which is phosphorylated with amidoxime in the 5' hydroxyl group of tromethamine. The third precursor drug, 6N hexyl fluorenyl TCN, was obtained from Dr. John Drach, University of Michigan. The structural formulas of the three precursor drugs are listed in Figure 1. 5, 〇· amidoxime-based triclopibine (A) and 5'0-amidino-mercaptophosphoric acid tricilide (B) The synthesis of the prodrug is described in the next section. The synthesis of 6N hexyl fluorenyl derivative (C) of TCN is described in Porcari et al. 5'0-Amidinoindolizine [6-Amino-4-mercapto-8-[5-decylamine decyl 20-((3-D-ribosefuranosyl)-pyrrolo[4] ,3,2-de])pyrimido[4,5-e]morphine] As shown in Figure 2 (next page), triclinbine (1 equivalent), n-Boc valine (1.2 equivalents) And DMAP (1.2 eq.) was dissolved in dry DMF and DCC (1.2 eq.) was added dropwise at room temperature in anhydrous DMF. The reaction mixture was stirred for 20 hours and then the solvent was evaporated to dryness. Amidoxime ester use 113 200902034 Rapid cerium oxide gel chromatography with 9: i DCM: methanol as the eluent and preparative HPLC to obtain the pure product. The structure was verified by Hi nmralC/Ms/MS. 5. Amidoxime esters are in equilibrium with 2, or 3, amidoxime esters, possibly as a result of the migration of the amino acid from the 5 position to the 2, and 3 positions. Based on 5 HPLC and H1 NMR analysis, about 85% to 90% of the lines were 5, S was present, and 10% to 15% of the product was 2' vinegar or 3' vinegar. 5'〇-绿绿醯-mercaptophosphoric acid trifluralin [6-amino-4-methyl-8_[5-[(nonylphenylphosphino) PNL-proline)]-( PD-ribosefuranosyl)-»bido[4,3,2-de]pyrimido[4,5-e]ratin]. Phenyl dichlorophosphate 10 (1 equivalent) and methyl guanidate hydrochloride (1 equivalent) were combined in anhydrous methylene chloride. Triethylamine (2 equivalents) was added to the mixture in anhydrous chloroform at -78 °C over 2 hours. The reaction mixture was allowed to return to room temperature and was further incubated for 20 hours. After the cultivation, the solvent was evaporated, and anhydrous diethyl ether was added to the residue. The resulting mixture was filtered under argon and the filtrate was dried to give (1). 15 This intermediate (5 eq.) was dissolved in dry THF and combined with N-methylimidazole (5 eq.) and added to EtOAc (1 eq.) in THF at -78. The reaction mixture was allowed to return to room temperature and stirred for additional 20 hours. The crude product linquied acid precursor was purified using rapid cerium oxide gel chromatography using: DCM:MeOH as eluent and preparative HPLC purification. 20 Cytotoxicity Cytotoxicity was tested in two systems, and HFF cells were visually inspected for 8 hours with compound culture, and cell survival assays were performed using cells that differentially expressed the cytotoxic AKT/PKB activity responsible for TCN. For HFF cells, TCNP and TCN 6N thiol derivatives were more toxic than TCN and confirmed histological studies in Dr. Dr. Dr. 114 200902034. Neither of these precursor drugs showed a cytotoxic effect (Table 3). fc.TCN, TCNP, and its individual precursor drugs are used in HFF fine makeup. The cytotoxicity was determined by visual inspection of HFF cells cultured for 8 days in the presence of various compounds in the concentration range of 100 μM. Table 3 Compound I £5〇iuM) TCN 100 TCNP 32 6N-mercapto-TCN 32 5'-0-Dileamine-based TCN &gt;100 5 '-0-ammonium-mercaptophosphoric acid TCN &gt;100 In the cell viability assay, TCN, TCNP, and 6Ν-mercapto-TCN 10 compounds are also very rare in human 549 cells, which are constitutively high Akt/PKB human non-small cell lung cancer cell lines and C〇1〇3 5 7 Or the pancreatic cancer cell line showing Akt expression (Table 4). In this Akt+ A459 cell, the compound showed cytotoxicity from 7.5 uM (TCN) to 70 uM (6N-mercapto-TCN); while in AV Colo357, none of the compounds had cytotoxicity (Table 4). 15 Table 4. TCNjiJCNP and let _TcN precursors in cell survival assays. The cytotoxicity of cytotoxicity was determined in two cell lines as described in the Methods section. 115 200902034 Table 4 Compound IC50 Α 549 (μΜ) IC50 at (: ο1ο357 (μΜ) TCN 7.5 &gt; 100 μΜ TCNP 27.0 &gt; 100 μΜ 6N-mercapto-TCN 70 &gt; 100 μΜ Therefore, according to structural formula and assay analysis The precursor drug shows unequal cytotoxicity. 5 The stability of the compound test compound in pH buffer 6.5 (intestinal pH) and liver homogenate. In Figure 9 and Figure 10, TCN and TCNP are known. And 5,-〇-delta-mercaptophosphoric acid triptriptone is stable in buffer and hepatic homogenate. But 5'-0- amidoxime-based tricilide prodrug (TCN-Vai) This is not the case. The prodrug showed a slight instability in the buffer and a significant instability in the liver homogenate. The liver homogenate was about 12 minutes (Fig. U and Fig. 12). The result was not unexpected. , 5, ester prodrugs in the physiological ? 11 and tissue homogenate is quite unstable 'and the prolyl linkage is considered to be more robust. TCN, TCNP and its precursors of intestinal 叨 t) 15 compound intestinal The channel absorption system was tested by injecting the compound into the interior of the rat duodenum. For comparison purposes, 3 mg of material was used for all compounds and plasma concentrations were determined 4 hours after administration. For TCN and TCN-N6-SI based alkyl compounds which are not completely soluble, a suspension of the material is injected into the duodenum. The results of these analyses are shown in Figures 13 through 17. 20 It can be seen that for TCN and TCNP (Figs. 13 and 14), the intestinal absorption of the compound is extremely small, and the peak plasma concentration is only about 125 Ng/ml. Phase 116 200902034 Inversely, the drug-drug compound (Figures 15-18) shows a much higher plasma concentration. The use of the three prodrugs resulted in a more complex Cp χ time curve because all of the prodrugs were at least partially hydrolyzed to their parent compound. For example, the Ν6-fluorenyl precursor has a peak concentration of 35 4 ng/ml 5 liters, but it is also not low at 1^1^5·2 Ng/ml). A similar pattern is also found in 5'0-Amidoxime-based Quercetin. 5, 〇-Amidoxime _ phosphatidyl citrate The prodrug showed maximum absorption, but in this case, the predominantly detectable compound was the parent compound TCNP, which had a Cmax of 173.6 ng/ml. The total exposure to TCN-containing compounds as measured by AUCG_4 hr also showed that the prodrugs had the greatest oral absorption potential before the link. These results are summarized in Table 5. Table 5. PK parameters for Cmax and AUC of TCN, TCNP and its individual 孓 孓 + + + ^ ^ ^

The Cmax value and the AUC value are determined by the Cp χ time curve shown in Fig. 13-17. Compound Cmax (Nike/ml) ^ AUC0^h7~^-i (Nike/milli)/hour TCN Below detection limit ----- ND TCNP 2.2 4.4+/-0.9 6N-醯-TCN --- Prodrug 35.4 102.1+/-5.5 TCN 5.2 _ 17.0+/-3.2 5Ό-Amidoxime-based tricilide prodrug 52.2 ------- 161+/-33.7 TCN 6.4 21.5+/- 4.8 Compound Cmax (NEK/ml) AUC0_4hr (Ng/ml)/hour 5Ό-Amidoxime-based guanidine citrate front peony 5.3 -- 17.4+/-2.5 TCNP 173.6 615.1+/-11.4 TCN 13.4 ------ 47.7+/-2.2 ---~~~~~, 117 200902034 Conclusion This feasibility study shows that TNC and TCNP have significant potential to develop oral drug products compared to their parental compounds. In particular, 5, 〇 缬 醯 醯 醯 醯 醯 醯 曲 曲 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得 可获得This result has implications for the development of these compounds towards oral TCN sputum repellents. The invention has been described with reference to preferred embodiments thereof. Variations and modifications of the present invention will be apparent to those skilled in the art from the Detailed Description of the Invention. All such changes and modifications are intended to be included within the scope of the present invention. 10 [Simplified Schematic Description] Fig. 1 shows the identification of API-2 (Quxiribin) identified as a candidate for Akt inhibitors in the NCI diversity set. Figure 丨a shows the chemical structure of API-2 (Quxi Libin). Figure 1B shows that API-2 inhibits the degree of AKT2 phosphorylation in AKT2 transduced NIH3T3 cells. Wil-type AKT2-transformed 15 NIH3T3 cells were treated with API-2 (ΙμΜ) for the indicated time and were subjected to immunoblotting analysis with anti-phospho-Akt-T308 and -S473 antibodies (top and bottom) . The figure below shows the performance of the total AKT2. In Figure 1C, three isomorphs of API-2 inhibition of Akt are shown. HEK293 cells were infected with HA-Akt1, HA-AKT2, and HA-AKT3 prior to EGF stimulation and treated with ΑΡΙ-2 (ΙμΜ) 20 or wortmannin (15 μΜ). Cells were lysed and anti-HA Antibody immunoprecipitation. The immunoprecipitate was subjected to in-vitro kinase assay (top) and immunoblot analysis with anti-phospho-Akt-T308 (bottom) antibody. The middle panel shows the performance of Aktl, AKT2, and AKT3 by transfer infection. Figure 1D shows that API-2 does not inhibit Akt in the tube. Compositional activity in vitro 118 200902034 The kinase assay for AKT2 recombinant protein in kinase buffer contains 1 μΜ API-2 (lane 3). Figure 2 demonstrates that API-2 does not inhibit closely related members of the PI3K, PDK1, and AGC kinase families. Figure 2A shows the PI3K kinase assay in vitro. HEK293 cells were serum-deficient and treated with API-2 (1 μΜ) or Watmanning (15 μΜ) for 30 minutes prior to EGF stimulation. The cells were lysed and immunoprecipitated with an anti-pll〇a antibody. The immunoprecipitate was assayed for in vivo kinase assay using ρι_4_ρ as the enzyme substrate. Figure 2 shows the effect of ΑΡΙ-2 on the activation of PDK1 in vitro (top panel). The filled circle shows inhibition by ΑΡΙ-2. The open circle 10 showed a 鞛% control of staurosporine inhibition, and Stromoxol was a potent PDK1 inhibitor (IC50 = 5 nM). The following figure shows the immunoblotting analysis of HEK293 cells. HEK293 cells were infected with Myc-PDK1 prior to EGF stimulation and treated with Watmanin or ΑΡΙ·2. The immune dot is detected by the indicated antibody. Figure 2C shows immunofluorescence of PKCa with phosphorylation of anti-phospho 15-pKCa-T638 (top panel) and total PKCa (bottom panel) antibodies followed by API-2 or non-selective PKC inhibitor R〇31 -8220 Point analysis. Figure 2D shows an in vitro SGK kinase assay. HEK293 cells were infected with HA-SGK transfer prior to EGF stimulation and treated with API-2 or Watmanin. The test tube kinase was immunoprecipitated with HA-SGK using MBP 20 as the enzyme substrate (top panel). The figure below shows the performance of HA-SGK transfected with infection. Figure 2E shows the results of the PKA stimuli assay. The immunopurified PKA is cultured in ADB buffer (Upstate Biotechnology Inc) containing a suitable inhibitor (API-2 or PKAI) and an enzyme substrate Kemptide. Quantify kinase activity. In the 2F figure, 119 200902034 shows the western ink dot. OVCAR3 cells were timed with API-2 treatment. The cell-dissolved product was immunized with an anti- scaly antibody (1 -4 map) and an anti-actin antibody (bottom panel). Figure 3 demonstrates that API-2 induces apoptosis by inhibiting 5 Akt activity and cell growth in human cancer cells with elevated Akt. Figure 3A shows the western blot after treatment with ΑΠ-2. The degree of phosphorylation of Akt is detected by anti-phospho-Akt-T308 antibody in a suitable human cancer cell line. The ink spots were again probed with anti-total Akt antibody (bottom panel). In Fig. 3B, a cell proliferation assay is shown. The cell lines indicated in the figure were treated with different doses of API-2 for 24 hours and 48 10 hours, and then analyzed by the CellTiter 96 Cell Proliferation Assay Kit (Promega). Figure 3C provides an analysis of apoptosis. Cells were treated with API-2, stained with annexin (v) and pi, and analyzed by FACScan. Figure 4 shows that in a mouse xenograft sheet, API 2 inhibits the downstream target of Akt in a cancer cell line with elevated Akt and has antitumor activity. In Figure 4A, it was verified that API-2 inhibited Akt phosphorylation of tuberin, Ba (j, AFX, and GSK-邛. After treatment with ΑΠ_2, 〇VCAR3 cells were lysed and immunized with applicable antibodies. Ink dot analysis. Figure 4B shows that API-2 can inhibit tumor growth. Tumor cells are injected into human nude mice with a low concentration of 20 Akt cells on the left side and a high concentration of Akt cells on the right side. When the tumor reaches an average of about 100-150 cubic millimeters. For large hours, animals were either loaded or treated with 丨mg/kg/day API-2. Each measurement represents an average of 1 tumor. Figure 4c shows with API-2 or vehicle (control) Representative images of mice treated with vcar3 (right) and OVCAR5 (left). Figure 4D shows an example of tumor size (bottom) and weight (top) at the end of the experiment 120 200902034. In Figure 4E, tumor dissolution The immunoblot analysis of the product used anti-phosphorus-Akt-S473 (top) and anti-AKT2 (under) in OVCAR-3-derived tumors treated with Api_2 (T3 and T4) and untreated (T1 and T2) Antibodies are carried out. 5 Figure 5 shows that ΑΠ-2 (Cucidine) inhibits Akt kinase activity in vitro The in-tube kinase assay was performed using a recombinant strain of PDK1 and Akt in a kinase buffer containing phospholipid-based myositol-3,4,5-P3 (PIP3), API-2 and tissue 脒H2B as an enzyme substrate. After a minute, the reaction was separated by SDS-PAGE and exposed to the membrane. 10 Figure 6a-6c provides the mRNA and amino acid sequence of human Aktl, also indicating the position of the restriction enzyme. Figures 7a-7d provide mRNA and amine of human Akt2. The base acid sequence also indicates the position of the restriction enzyme. Figures 8a-8c provide the mRNA and amino acid sequence of human Akt3, also indicating the position of the 15 restriction enzyme. Figure 9 shows TCN, TCNP and 5'-0-guanamine Buffer stability of mercaptophosphoric acid triterpene (TCNP_Val.) Figure 10 shows TCN, TCNP and 5'-0- amidoxime phosphoryl fluoxetine (TCNPJVal.) Stability in liver homogenate. 20 Figure 11 shows the buffer stability of 5,-0- amidoxime-t-civiline (TCN-Val). Figure 12 shows the 5,-0- amidoxime group. The stability of tricycline (TCN-Val) in liver homogenate.

Figure 13 shows the plasma concentrations of TCN 121 200902034 and TCNP after administration of 3 mg TCN (n = 3) via the duodenum. Figure 14 shows the plasma concentrations of TCN and TCNP after administration of 3 mg of TCN (n=3) via the duodenum. Figure I5 shows the plasma concentration of 5 TCN, TCNP and 6N TCN hexyl fluorenyl derivative (TCN_1436) after administration of 3 mg of TCN_1436 (n=2) in the duodenum. Figure 16 shows the plasma concentrations of TCN, TCNP and TCN-Val after administration of 3 mg of 5'-0-amidoxime tromethamine (TCN-Val) (n = 3) via the duodenum. 10 Figure 17 shows plasma of TCN, TCNP and TCN-Val after administration of 3 mg of 5'-0-amidoximephosphoric acid triclinide (TCNP-Val) (n=3) via the duodenum. concentration. [Main component symbol description] (none) 122

Claims (1)

200902034 X. Patent application scope: 1. A compound of the formula: Each of Ri and R2 is -H, and 5 R2', R3', and R5' are each -H or an amino acid. 2. - Compounds of the following formula: 1^ and 112 are each -H; and R2', R3', and R5' are each -H, optionally substituted 10 phosphate or phosphonate (including monophosphate, diphosphate or triphosphate) Or stabilized phosphate precursors). 3. - Compounds of the following formula: 123 200902034 Production 3 R3 is a linear crc18 alkyl group, and R2', R3', and R5' are each -Η. A pharmaceutical composition comprising a compound 5 as claimed in claim 1 and a pharmaceutically acceptable carrier. 5. A pharmaceutical composition comprising a compound as claimed in claim 2 and a pharmaceutically acceptable carrier. 6. A pharmaceutical composition comprising a compound as claimed in claim 3 and a pharmaceutically acceptable carrier. 10 7. The pharmaceutical composition of claim 4, which is suitable for oral administration. 8. The pharmaceutical composition of claim 5, which is suitable for oral administration. 9. The pharmaceutical composition of claim 6 is suitable for oral administration. 10. The pharmaceutical composition of claim 4, wherein the compound is present in an amount from about 0" to about 1000 mg. 11. The pharmaceutical composition of claim 5, wherein the compound is present in an amount from about 0.1 ng to about 1000 mg. 124 200902034 12. The pharmaceutical composition of claim 6, wherein the compound is present in an amount from about 0.1 ng to about 1000 mg. 13. A method of treating a neoplasm or cancer in a mammal comprising administering to a mammal in need thereof an effective amount of a compound as claimed in claim 1 of the patent. 14. A method of treating a tumor or cancer in a mammal comprising administering to a mammal in need thereof an effective amount of a compound as in claim 2 of the patent application. 15. A method of treating a tumor or cancer in a mammal comprising administering to a mammal in need thereof an effective amount of a compound as in claim 3 of the patent application. 125