WO2007095389A2

WO2007095389A2 - Treatment of hyperproliferative diseases with camptothecine n-oxide and analogs

Info

Publication number: WO2007095389A2
Application number: PCT/US2007/004253
Authority: WO
Inventors: John G. Curd; John F. W. Keana; Alshad S. Lalani; Paul B. Westberg; Bradford S. Goodwin
Original assignee: Novacea, Inc.
Priority date: 2006-02-17
Filing date: 2007-02-20
Publication date: 2007-08-23
Also published as: WO2007095389A3

Abstract

The invention relates to N-oxides of camptothecin analogs having activity for treating hyperproliferative disorders. Pharmaceutical compositions comprising therapeutically effective amount of camptothecin analog N-oxides and bis N-oxides, or a pharmaceutically acceptable salt or prodrug thereof, are disclosed. Further, the invention relates to methods of using the compounds, alone or in combination with one or more other active agents or treatments, to treat hyperproliferative disorders.

Description

TREATMENT OF HYPERPROLIFERATTVE DISEASES WITH CAMPTOTHECIN N-OXIDE AND ANALOGS

BACKGROUND OF THE INVENTION

Field of the Invention

[0001] The present invention relates to camptothecin analog N-oxides having activity for treating hyperproliferative disorders. Further, the invention relates to methods of using the compounds, alone or in combination with one or more other active agents or treatments, to treat hyperproliferative disorders.

Related Art

[0002] One in every four deaths in the United States is due to cancer, and cancer is the second leading cause of death. U.S. Cancer Statistics Working Group; United States Cancer Statistics: 1999-2001 Incidence, Atlanta (GA): Department of Health and Human Services, Centers for Disease Control and Prevention, and National Cancer Institute (2004). The National Cancer Institute reports that almost 10 million Americans have a history of invasive cancer, while the American Cancer Society estimates that in the year 2004, over 1.3 million Americans will receive a diagnosis of invasive cancer with over a half million cases resulting in death. American Cancer Society, Cancer Facts & Figures 2004. These statistics exclude the 1 million cases of basal and squamous cell skin cancers that are expected to be diagnosed in the United States.

[0003] Cancers are classified based on the organ and cell tissue from which the cancer originates, including: (i) carcinomas (most common kind of cancer which originates in epithelial tissues, the layers of cells covering the body's surface or lining internal organs and various glands); (ii) leukemias (origination in the blood-forming tissues, including bone marrow, lymph nodes and the spleen); (iii) lymphomas (originates in the cells of the lymph system); (iv) melanomas (originates in the pigment cells located among the epithelial cells of the skin); and (v) sarcomas (originates in the connective tissues of the body, such as bones, muscles and blood vessels). {See Molecular Biology of the Cell: Third ^"Edition, "Cancer," Chapter 24, pρ.1255-1294, B. Alberts et ah, (eds.), Garland Publishing, Inc., New York (1994); and Stedman's Pocket Medical Dictionary; Williams and Wilkins, Baltimore (] 987)). Within these broad cancer classifications, there are over one hundred cancer subclassifications, such as breast, lung, pancreatic, colon, and prostate cancer, to name a few.

[0004] Cancer cells develop as a result of damage to a cell's DNA {i.e., altered

DNA sequence or altered expression pattern) from exposure to various chemical agents, radiation, viruses, or when some not-yet-fully-understood internal, cellular signaling event occurs. Most of the time when a cell's DNA becomes damaged, the cell either dies or is able to repair the DNA. However, for cancer cells, the damaged DNA is not repaired and the cell continues to divide, exhibiting modified cell physiology and function.

[0005] Neoplasms, or tumors, are masses of cells that result from an aberrant, accelerated rate of growth {i.e., hyperproliferative cell growth). As long as the tumor cells remain confined to a single mass, the tumor is considered to be benign. However, a cancerous tumor has the ability to invade other tissues and is termed malignant. In general, cancer cells are defined by two heritable properties: the cells and their progeny 1) reproduce in defiance of normal restraints, and 2) invade and colonize the territories of other cells.

(0006] Cancerous tumors are comprised of a highly complex vasculature and differentiated tissue. A large majority of cancerous tumors have hypoxic components, which are relatively resistant to standard anti-cancer treatment, including radiotherapy and chemotherapy. Brown, Cancer Res. 59:5863 (1999); and Kunz, M. et al, MoI. Cancer 2:1 (2003). Thomlinson and Gray presented the first anatomical model of a human tumor that describes a 100 to 150 μm thick hypoxic layer of tissue located between the blood vessels and necrotic tumor tissues. 10007] Research has shown that the hypoxic tissues within a number of cancerous tumors promote the progression of the cancer by an array of complex mechanisms. See, Brown., supra, and Kunz et al., supra. Among these are activation of certain signal transduction pathways and gene regulatory mechanisms, induction of selection processes for gene mutations, tumor cell apoptosis and tumor angiogenesis. Most of these mechanisms contribute to tumor progression. Therefore, tissue hypoxia has been regarded as a central factor for tumor aggressiveness and metastasis. Therapies that target hypoxic tissues within a tumor would certainly provide improved treatments to patients suffering from tumor-related cancers and/or disorders.

[0008] In addition to cancer, there exist a number of hyperproliferative diseases and/or disorders that are associated with the onset of hypoxia in a given tissue. For example, Shweiki et al. explain that inadequate oxygen levels often lead to neovascularization in order to compensate for the needs of the hypoxic tissue. Neovascularization is mediated by expression of certain growth factors, such as vascular endothelial growth factor (VEGF). Shweiki et al., Nature 359:843 (1992). However, when certain tissues or growth factors are either directly or indirectly upregulated in response to hypoxia without sufficient feedback mechanisms for controlling tissue expression, various diseases and/or disorders may ensue (i.e., by^' hypoxia-aggravated hyperproliferation).

[0009] Camptothecin is a naturally occurring cytotoxic alkaloid which is known to inhibit the enzyme topoisomerase I and is a potent anti-tumor agent. Camptothecin was isolated from the wood and bark of Camptotheca acuminata. See U.S. patent 5,932,588.

[0010] U.S. Patent No. 6,350,756 describes a group of camptothecin analogs having the following structure:

wherein R is Ri -O-(CH₂)_m-, m is an integer of 1-10 (preferably 1-5); and

Ri is lower alkyl; phenyl optionally substituted with from one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, formyl, lower alkyl carbonyl, hydroxycarbonyl, lower alkylcarbonyloxy, benzyloxy, optionally substituted piperazino, lower alkoxycarbonyl, and lower alkylcarbonylamino; cycloalkyl of 3-7 carbons, optionally substituted with one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino; a fused, 2-, 3-, or 4-riήg heterocyclic system optionally substituted with one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino;

1- or 2-naphthyl optionally substituted with from one to four substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonyl amino; a 5 or 6 membered heterocyclic ring containing one or two nitrogen atoms, which ring is optionally substituted with one or two substituents selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino ;

R₂ is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, RC(O)O (R defined hereinbefore), cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, -C(O)H, lower alkoxycarbonyl, tri-lower alkylsilyl, lower alkylcarbonyloxy, lower alkylcarbonylamino, lower alkylcarbonyloxymethyl, substituted vinyl, 1 -hydroxy-2-nitroethyl, alkoxycarbonylethyl, aminocarbonyl, mono- or di-alkylcarbonyl, alkylcarbonyloxymethyl, benzoyl methyl, benzylcarbonyloxymethyl, or mono- or di-lower alkoyxymethyl.

R₃ is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, RC(O)O (R defined hereinbefore) cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, CH₂NR₇Rs (where each of R₇ and Rs is independently H-, alkyl of 1-6 carbons, optionally substituted phenyl, hydroxy lower alkyl, amino lower alkyl, or mono- or dialkylamino lower alkyl-, or R₇ and Rs taken together with -N- represent a cyclic amino-), -C(O)H, CH2R₉ (where R₉ is lower alkoxy, CN, amino lower alkoxy, mono- or di-lower alkylamino lower alkoxy, lower alkylthio, amino lower alkylthio, or mono- or di-lower alkylamino lower alkylthio), or NR₁₀R_{1 1} (where each of Rio and Rn is independently hydrogen, lower alkyl, phenyl, hydroxy lower alkyl, amino lower alkyl, or mono- or di- lower alkyl, or Rio and Rn taken together with -N- represent a cyclic amino), dialkylamino alkyl, lower alkylcarbonyloxy, or lower alkylcarbonylamino; and R4 is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, RC(O)O (R defined hereinbefore) cyano, nitro, amino, amino lower alkyl, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, carbamoyloxy, lower alkylcarbonyloxy, or lower alkylcarbonylamino, or R₄ together with R₃ is methylenedioxy;

R₅ is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, RC(O)O (R defined hereinbefore), cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, or lower alkylcarbonylamino; and

Re is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, RC(O)O (R defined hereinbefore), cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxcarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, or lower alkylcarbonylamino.

[0011] Among camptothecin analogs disclosed in U.S. Patent No. 6,350,756 are P-dimethylaminomethyl-lO-hydroxycamptothecin (topotecan),

V-ethyl-lOf^l-piperidino^l-piperidinoJcarbonyloxycamptothecin (irinotecan), 9-aminocamptothecin, 10-aminocamptothecin,

10, 1 1 -methyl enedioxycamptothecin and 7-ethyl- 10-hyrdoxycamptothecin (SN-38) having (20S) configuration.

[0012] Rahier N. J. et al. (Org. Lett. 6(3):32l-24 (2004)) also reported camptothecin esterifled with water solubilizing groups such RPO₃H₂ wherein R is OH, OCH₃, CH₃ or PH.

BRIEF SUMMARY OF THE INVENTION

[0013] The present invention is related to compounds, pharmaceutical compositions and methods for treating hyperproliferative disorders, such, as cancer and inflammation. One aspect of the invention is drawn to compounds having Formula I:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

R' is hydrogen, Ri-O-(CH₂)_m-CO- or Ri₆PO₂H-, m is an integer of 0-10 (preferably 1-5); and

Ri is lower alkyl; phenyl optionally substituted with from one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, formyl, lower alkyl carbonyl, hydroxycarbonyl, lower alkylcarbonyloxy, benzyloxy, optionally substituted piperazino, lower alkoxycarbonyl, and lower alkylcarbonylamino; cycloalkyl of 3-7 carbons, optionally substituted with one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower- alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino; a fused, 2-, 3-, or 4-ring heterocyclic system optionally substituted with one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino; 1- or 2-naphthyl optionally substituted with from one to four substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino; a 5 or 6 membered heterocyclic ring containing one or two nitrogen atoms, which ring is optionally substituted with one or two substituents selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino;

R₂ is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, R¹C(O)O (R' defined hereinbefore), cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, -C(O)H, lower alkoxycarbonyl, tri-lower alkylsilyl, lower alkylcarbonyloxy, lower alkylcarbonylamino, lower alkylcarbonyloxymethyl, substituted vinyl, 1 -hydroxy-2-nitroethyl, alkoxycarbonylethyl, aminocarbonyl, mono- or di-alkylcarbonyl, alkylcarbonyloxymethyl, benzoylmethyl, benzylcarbonyloxymethyl, or mono- or di-lower alkoyxymethyl.

R3 is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, R¹C(O)O (R' defined hereinbefore) cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, CH₂NR₇RgRi ₂ (where each of R₇ and R₈ is independently H-, alkyl of 1-6 carbons, optionally substituted phenyl, hydroxy lower alkyl, amino lower alkyl, or mono- or dialkylamino lower alkyl, or R₇ and Rg taken together with -N- represent a cyclic amino-), -C(O)H, -CH2R9 (where R₉ is lower alkoxy, CN, amino lower alkoxy, mono- or di-lower alkylamino lower alkoxy, lower alkylthio, amino lower alkylthio, or mono- or di-lower alkylamino lower alkylthio), or NR₁₀R_{1 1}R₁₂ (where each of Rio and Rn is independently hydrogen, lower alkyl, phenyl, hydroxy lower alkyl or amino lower alkyl, or Rio and Rn taken together with -N- represent a cyclic amino), dialkylamino alkyl, lower alkylcarbonyloxy, or lower alkylcarbonylamino; and

R₄ is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, amino lower alkyl, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, carbamoyloxy, lower alkylcarbonyloxy, lower alkylcarbonylamino or RnCOO- wherein Ri₇ is a 5 or 6 membered heterocyclic ring containing one or two nitrogen atoms and substituted with one or two substituents selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, NRi₀RnRi₂ (where each of Rio and Ru is independently H-, lower alkyl, phenyl, hydroxy lower alkyl, amino lower alkyl, or Rio and Rn taken together with -N- represent a cyclic amino-), lower alkylcarbonyloxy, or lower alkylcarbonylamino; or R₄ together with R₃ is methylenedioxy;

R₅ is hydrogen, halo, lower alkyl, lower, alkoxy, hydroxy, R¹C(O)O (R' defined hereinbefore), cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, or lower alkylcarbonylamino; and

Rβ is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, R¹C(O)O (R¹ defined hereinbefore), cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxcarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, or lower alkylcarbonylamino;

Ri₆ is hydroxy, alkyl, alkoxy or phenyl; and each of R₁₂ and Rj ₅ is O or is absent, provided that at least one OfR]₂ and R₁₅ is O. In one embodiment, the compounds having formula I are N_t-oxides of camptothecin analogs having both the 20(RS) and 20(S) configurations. In another embodiment, the compounds having formula I are bis N-oxides, one of the N-oxide oxygen atoms being located on Ni and the other N-oxide oxygen atom being located on a tertiary amino group on a side chain. The camptothecin analog N-oxide and bis N-oxide compounds of the present invention are selected from the group consisting of: (7-ethyl- 10-[4-(l-piperidino)- 1 -piperidino]carbonyloxy)-camptothecin (irinotecan);

9-nitro camptothecin;

7-chloro camptothecin;

7-n-propyldimethylsilyl camptothecin;

10-hydroxy-7-n-propyldimethylsilyl camptothecin; lO-hydroxy-7-chlorocamptothecin;

10-acetoxy-7-n-propyldimethylsilyl camptothecin;

10-acetoxy-7-chlorocamptothecin;

7-tert-butyldimethylsilyl camptothecin;

10-hydroxy-7-tert-butyldimethylsilyl camptothecin;

1 O-acetoxy-7-tert-butyldimethylsilyl camptothecin;

9-hydroxy camptothecin;

9-amino camptothecin;

10-amino camptothecin;

9-amino- 10-hydroxy camptothecin;

9-amino- 10, 11 -methylenedioxy camptothecin;

9-methylamino camptothecin;

9-methyl camptothecin;

9-dimethylaminomethyl camptothecin;

9-chloro camptothecin;

9-fluoro camptothecin;

9-piperidino camptothecin;

9-dimethylaminomethyl- 10-hydroxy camptothecin (topotecan);

9-morpholinomethyl camptothecin;

10-hydroxy camptothecin;

9,10-dichloro camptothecin;

10-bromo camptothecin;

10-chloro camptothecin;

10-methyl camptothecin;

10-fluoro camptothecin; 10-nitro camptothecin; 10,11 -methyl enedioxy camptothecin; 10-formyl camptothecin; 10-nonylcarbonyloxy camptothecin; 10-undecylcarbonyloxy camptothecin; 10-pentadecylcarbonyloxy camptothecin; 10-heptadecylcarbonyloxy camptothecin; 10-nonadecylcarbonyloxy camptothecin; 9-nitro-10,l 1 -methylenedioxy camptothecin; 9-(4-methylpiperazinylmethyl)- 10-hydroxy camptothecin; 9-[4-(l -piperidino)- 1-piperidinomethyl]- 10-hydroxy camptothecin; 9-methyl-l 0,11 -methylenedioxy camptothecin; 9-chloro-10,l 1 -methylenedioxy camptothecin; 9-cyano-10,l 1 -methylenedioxy camptothecin; 9-acetoxy-10,l 1 -methylenedioxy camptothecin; 9-acetylamino-10,l 1 -methylenedioxy camptothecin; 9-aminomethyl- 10-hydroxy camptothecin; 9-ethoxymethyl- 10-hydroxy camptothecin; 9-methylaminomethyl-l 0-hydroxy camptothecin; 9-n-propylaminomethyl-l 0-hydroxy camptothecin; 9-dimethylaminomethyl-l 0-hydroxy camptothecin; 9-cyclohexylaminomethyl-l 0-hydroxy camptothecin; 9-(2-hydroxyethyl)aminomethyl-10-hydroxy camptothecin; 9-(trimethylammonio)methyl- 10-hydroxy camptothecin, methanesulfonate;

9-morpholinomethyl-l 0-hydroxy camptothecin;

9-cyanomethyl- 10-hydroxy camptothecin; camptothecin-7-aldehyde;

10-methoxy camptothecin-7-aldehyde;

7-acetoxymethyl camptothecin;

7-acetoxym ethyl- 10-methyl camptothecin; 7-cyano-10-methoxy camptothecin;

7-cyano camplothecin;

7-formylethenyl camptothecin;

7-ethoxycarbonylethenyl camptothecin;

7-cyanoethenyl camptothecin;

7-(2,2-dicyanoethenyl) camptothecin;

7-(2-cyano-2-ethoxycarbonyl)ethenyl camptothecin;

7-ethoxycarbonylethyl camptothecin;

7-ethyl camptothecin;

7-n-propyl camptothecin;

7-acetoxymethyl camptothecin;

7-n-ρropylcarbonyloxymethyl camptothecin;

7-ethoxycarbonyl camptothecin;

7-ethyl-10-hydroxy camptothecin (SN-38);

7-ethyl-lO-acetyloxy camptothecin;

7 -methyl- 10-aminocarbonyloxy camptothecin;

7-n-propyl-10-piperidinocazbonyloxy camptothecin;

7-ethyl- 10-(2-dimethylamino)ethyl camptothecin;

7-ethyl-10-[4(l-piperidino)-piperidino carbonyloxy camptothecin;

7-ethyl- 10-( 1 -piperazine)carbonyloxy camptothecin;

7-ethyl- 10-(4-i-propylaminocarbonylmethylpiperazine)carbonyloxy camptothecin;

7-ethyl- 10-[4( 1 -pyrrolidinyl)piperazine]carbonyloxy camptothecin ;

7-ethyl-10-[(4-(dimethylamino)-l-piperidino]carbonyloxy camptothecin;

7-ethyl-10-[4-(di-n-propylamino)-l-piperidinol]carbonyloxy camptothecin;

7-ethyl- 10-[(4-(di-n-butyl amino)- 1 -piperidinojcarbonyloxy camptothecin;

7-ethyl-10-[4-(l-pyrrolidino)-l-piperidino)]carbonyloxy camptothecin; 7-ethyl-10-[4-(l-piperidino)-l-piperidino]carbonyloxy camplothecin; and

7-ethyl-10-[N-mcthyl-N-2-(dimethylamino)ethylamino]carbonyloxy camptothecin.

[0015] According to another aspect of the invention, a therapeutically effective amount of a compound having Formula I is provided in the form of a pharmaceutical composition having at least one pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition comprises a therapeutically effective amount of an N-oxide of camptothecin or camptothecin analog. In another embodiment, the camptothecin analog is selected from the group consisting of 9-dimethylaminomethyl- 10-hydroxycamptothecin N-oxide (topotecan N-oxide),

9-dimethylaminomethyl-lO-hydroxycamptothecin bis N-oxide (topotecan bis N-oxide), 7-ethyl-10[4-(l-piperidino)-l-piperidino]carbonyloxycamptothecin N-oxide (irinotecan N-oxide), 7-ethyl-10[4-(l-piperidino)- l-piperidino]carbonyloxycamptothecin bis N-oxide (irinotecan bis N-oxide), 9-aminocamptothecin N-oxide, 10-aminocamptothecin N-oxide, 10,11-methylenedioxycamptothecin N-oxide and 7-ethyl-10- hyrdoxycamptothecm N-oxide (SN-38 N-oxide). ^•

[0016] Another aspect of the present invention is related to methods for treating hyperproliferative disorders. In certain aspects of the invention, the hyperproliferative disorder is cancer. In one embodiment, the cancer is a solid tumor. In another embodiment, the cancer is selected from the group consisting of colon cancer, brain cancer, glioma, multiple myeloma, head and neck cancer, hepatocellular cancer₅ melanoma, ovarian cancer, cervical cancer, renal cancer, and non-small cell lung cancer. In a further embodiment, the cancer is acute and chronic lymphocytic leukemia, acute granulocytic leukemia, adrenal cortex carcinoma, bladder carcinoma, breast carcinoma, cervical carcinoma, cervical hyperplasia, choriocarcinoma, chronic granulocytic leukemia, chronic lymphocytic leukemia, colon carcinoma, endometrial carcinoma, esophageal carcinoma, essential thrombocytosis, genitourinary carcinoma, hairy cell leukemia, head and neck carcinoma, Hodgkin's disease, Kaposi's sarcoma, lung carcinoma, lymphoma, malignant carcinoid carcinoma, malignant hypercalcemia, malignant melanoma, malignant pancreatic insulinoma, medullary thyroid carcinoma, melanoma, multiple myeloma, mycosis fungoides, myeloid and lymphocytic leukemia, neuroblastoma, non-Hodgkin's lymphoma, osteogenic sarcoma, ovarian carcinoma, pancreatic carcinoma, polycythemia vera, primary brain carcinoma, primary macroglobulinemia, prostatic carcinoma, renal cell carcinoma, rhabdomyosarcoma, skin cancer, small-÷cell lung carcinoma, soft-tissue sarcoma, squamous cell carcinoma, stomach carcinoma, testicular carcinoma, thyroid carcinoma, or Wilms' tumor.

[0017] In further aspects of the invention the hyperproliferative disorder is any one of age-related macular degeneration, Crohn's disease, cirrhosis, chronic inflammatory-related disorders, proliferative diabetic retinopathy, proliferative vitreoretinopathy, retinopathy of prematurity, granulomatosis, immune hyperproliferation associated with organ or tissue transplantation, an immunoproliferative disease or disorder, e.g., inflammatory bowel disease, psoriasis, rheumatoid arthritis, systemic lupus erythematosus (SLE), vascular hyperproliferation secondary to retinal hypoxia, or vasculitis.

[0018] In one embodiment the invention is drawn to methods of treating, ameliorating, or preventing hyperproliferative disease in a subject comprising administering to said subject a therapeutically effective amount of an N-oxide of camptothecin or analog thereof. In another embodiment, the camptothecin analog is selected from the group consisting of 9-dimethylaminomethyl- lOrhydroxycamptothecin N-oxide (topotecan N-oxide),

9-dimethylaminomethyl-lO-hydroxycamptothecin bis N-oxide (topotecan bis N-oxide), 7-ethyl-10[4-(l-piperidino)-l-piperidino]carbonyloxycamptothecin N-oxide (irinotecan N-oxide), 7-ethyl-10[4-(l-piperidino)- l-piperidiπo]carbonyloxycamptothecin bis N-oxide (irinotecan bis N-oxide), 9-aminocamptothecin N-oxide, 10-aminocamptothecin N-oxide, 10,11-methylenedioxycamptothecin • N-oxide and 7-ethyl-10- hyrdoxycamptothecin N-oxide (SN-38 N-oxide).

[0019] An additional aspect of the present invention is a method for treating, ameliorating, or preventing hyperproliferative disorders in an animal comprising administering to the animal a therapeutically effective amount of a compound having Formula I in combination with one or more active agents or treatments. In one embodiment, the one or more active agent or treatment is a chernotherapeutic agent, a radiotherapeutic agent/treatment, an anti-angiogenesis agent, a vascular targeting agent, a hypoxia-inducible factor 1 (HIFl) inhibitor, an Hsp90 inhibitor, a tyrosine kinase inhibitor, a serine/threonine kinase inhibitor, a proteasome inhibitor, an HDAC inibitor, a caspase inducer, a CDK inhibitor, and a proapoptotic molecule. In another embodiment, the one or more active agent or treatment is used, has been used, or is known to be useful for the treatment of the hyperproliferative disorder.

[0020] In one embodiment, the method of treating, ameliorating, or preventing hyperproliferative disorder in an animal comprises administering to the animal a therapeutically effective amount of camptothecin N-oxide or bis N-oxide or analog thereof. In particular embodiments, the camptothecin analog N-oxide or bis N-oxide is selected from the group consisting of P-dimethylaminomethyl-lO-hydroxycamptothecin N-oxide (topotecan N-oxide) P-dimethylaminomethyl-lO-hydroxycamptothecin bis N-oxide (topotecan bis N-oxide), 7-ethyl-10[4-(l-piperidino)- l-piperidino]carbonyloxycamptothecin N-oxide (irinotecan N-oxide), 7-ethyl-10[4-(l-piperidino)-l-piperidino]carbonyloxycamptothecin bis

N-oxide (irinotecan bis N-oxide), 9-aminocamptothecin N-oxide, 10-aminocamptothecin N-oxide, 10,11-methylenedioxycamptothecin N-oxide and 7-ethyl-lO-hyrdoxycamptothecin N-oxide (SN-38 N-oxide) in combination with one or more active agents or treatments, for example, chemotherapeutic agents or radiotherapeutic agents/treatments.

[0021] In preferred embodiments of the invention, the one or more chemotherapeutic agents can be any chemotherapeutic agent which is used, has been used, or is known to be useful for the treatment of hyperproliferative disorders.

[0022] In preferred embodiments of the invention, the one or more radiotherapeutic agents or treatments can be external-beam radiation therapy, brachytherapy, thermotherapy, radiosurgery, charged-particle radiotherapy, neutron radiotherapy, photodynamic therapy, or radionuclide therapy.

[0023] In one embodiment of the invention, the compound having Formula T can be administered prior to, during, and/or beyond administration of the one or more chemotherapeutic agents or radiotherapeutic agents or treatments. In another embodiment of the invention, the method of administering a compound having Formula I in combination with one or more chemotherapeutic agents or radiotherapeutic agents or treatments is repeated more than once.

[0024] The combination of a compound having Formula I and one or more chemotherapeutic agents or radiotherapeutic agents or treatments of the present invention will have additive potency or an additive therapeutic effect. The invention also encompasses synergistic combinations where the therapeutic efficacy is greater than additive. Preferably, such combinations will reduce or avoid unwanted or adverse effects. In certain embodiments, the combination therapies encompassed by the invention will provide an improved overall therapy relative to administration of a compound having Formula ϊ or any chemotherapeutic agent or radiotherapeutic agent or treatment alone. In certain embodiments, doses of existing or experimental chemotherapeutic agents or radiotherapeutic agents or treatments will be reduced or administered less frequently which will increase patient compliance, thereby improving therapy and reducing unwanted or adverse effects.

[0025] Further, the methods of the invention will be useful not only with previously untreated patients but also will be useful in the treatment of patients partially or completely refractory to current standard and/or experimental cancer therapies, including but not limited to radiotherapies, chemotherapies, and/or surgery. In a preferred embodiment, the invention will provide therapeutic methods for the treatment or amelioration of hyperproliferative disorders that have been shown to be or may be refractory or non-responsive to other therapies.

[0026] While not wishing to be bound by any theory, it is believed that some of the N-oxide compounds of the invention will function as prodrugs with greatly diminished cytotoxicity. It is believed that these N-oxide compounds will be activated under hypoxic conditions within the target tissues (i.e., reduced at the nitrogen atom), followed by, for example, intercalation with DNA and/or inhibition of DNA topoisomerase I, diminishing cells' ability to replicate. Other N-oxide compounds of the invention may have intrinsic cytotoxic activity. Since a number of pathological tissues have significant hypoxic components which promote hyperproliferation, it is believed that this portion of tissue will be preferentially targeted.

DETAILED DESCRIPTION OF THE INVENTION

[0027] One aspect of the invention is drawn to compounds having Formula I:

I or a pharmaceutically acceptable salt or prodrug thereof, wherein: R¹, R₂-R₆ and Rj₅ are as defined above. [0028] Another aspect of the invention is drawn to the compound of Formula π:

π or a pharmaceutically acceptable salt or prodrug thereof, wherein R', R₂, R₄ and Ri ₅ are as defined above. [0029] Another aspect of the invention is drawn to the compound of Formula in:

in or a pharmaceutically acceptable salt or prodrug thereof, wherein R' is hydrogen, R₂, R₄ and R₁₅ are as defined above.

[0030] According to another aspect of the invention, a therapeutically effective amount of a compound having Formula I, or a pharmaceutically acceptable salt thereof, and at least one other active agent is provided in the form of a pharmaceutical composition having at least one pharmaceutically acceptable carrier. In certain instances, the at least one other active agent is a chemotherapeutic agent (including an active vitamin D compound). Compounds having Formula I may be formulated in a single formulation with the other active agent(s), or formulated independently.

[0031] According to one aspect of the invention, methods for treating, ameliorating, or preventing hyperproliferative disorders are provided, wherein a therapeutically effective amount of a compound having Formula I, or a pharmaceutically acceptable salt thereof, is administered to an animal in need thereof. In certain aspects of the invention, the hyperproliferative disorder is cancer.

[0032] A further aspect of the invention relates to methods for treating, ameliorating, or preventing a hyperproliferative disorder comprising administering a therapeutically effective amount of a compound having Formula I, or a pharmaceutically acceptable salt thereof, in combination with at least one other active agent or treatment to a patient in need thereof. In certain embodiments, combinations of a compound having Formula I with a chemotherapeutic agent are administered. In one embodiment, the chemotherapeutic agent is selected from gemcitabine and irinotecan.

[0033] Hyperproliferative disorders which can be treated with the compounds having Formula I include any hypoxia-aggravated hyperproliferative disease and/or disorder, such as any number of cancers. Generally, such cancers include, without limitation, cancers of the bladder, brain, breast, cervix, colon, endometrium, esophagus, head and neck, kidney, larynx, liver, lung, oral cavity, ovaries, pancreas, prostate, skin, stomach, and testis. Certain of these cancers may be more specifically referred to as acute and chronic lymphocytic leukemia, acute granulocytic leukemia, adrenal cortex carcinoma, bladder carcinoma, breast carcinoma, cervical carcinoma, cervical hyperplasia, choriocarcinoma, chronic granulocytic leukemia, chronic lymphocytic leukemia, colon carcinoma, endometrial carcinoma, esophageal carcinoma, essential thrombocytosis, genitourinary carcinoma, hairy cell leukemia, head and neck carcinoma, Hodgkin's disease, Kaposi's sarcoma, lung carcinoma, lymphoma, malignant carcinoid carcinoma, malignant hypercalcemia, malignant melanoma, malignant pancreatic insulinoma, medullary thyroid carcinoma, melanoma, multiple myeloma, mycosis fungoides, myeloid and lymphocytic leukemia, neuroblastoma, non-Hodgkin's lymphoma, osteogenic sarcoma, ovarian carcinoma, pancreatic carcinoma, polycythemia vera, primary brain carcinoma, primary macroglobulinemia, prostatic carcinoma, renal cell carcinoma, rhabdomyosarcoma, skin cancer, small-cell lung carcinoma, soft-tissue sarcoma, squamous cell carcinoma, stomach carcinoma, testicular carcinoma, thyroid carcinoma, and Wilms¹ tumor. In one embodiment, the cancer is a solid tumor. In another embodiment, the cancer is selected from the group consisting of colon cancer, brain cancer, glioma, multiple myeloma, head and neck cancer hepatocellular cancer, melanoma, ovarian cancer, cervical cancer, renal cancer, and non-small cell lung cancer.

[0034] Animals which may be treated according to the present invention include all animals which may benefit from administration of compounds having Formula I. Such animals include humans, pets such as dogs and cats, and veterinary animals such as cows, pigs, sheep, goats and the like.

[0035] The term "alkyl" refers to a monovalent, saturated aliphatic hydrocarbon radical having the indicated number of carbon atoms. For example, a "C 1-6 alkyl" or an "alkyl of 1-6 carbons" or "AIk 1-6" would refer to any alkyl group containing one to six carbons in the structure. "C 1-20 alkyl" refers to any alkyl group having one to twenty carbons. Alkyl may be a straight chain (i.e. linear) or a branched chain. Lower alkyl refers to an alkyl of 1-6 carbons. Representative examples lower alkyl radicals include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, isopentyl, amyl, sec-butyl, tert-butyl, tert-pentyl and the like. Higher alkyl refers to alkyls of seven carbons and above. These include n-heptyl, n-octyl, n-nonyl, n-decyl, n- dodecyl, n-tetradecyl, n-hexadecyl, n-octadecyl, n-eicosyl, and the like, along with branched variations thereof. The radical may be optionally substituted with substituents at positions that do not significantly interfere with the preparation of compounds falling within the scope of this invention and that do not significantly reduce the efficacy of the compounds. The alkyl is optionally substituted with one to five substituents independently selected from the group consisting of halo, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alky!, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino. f 0036] The term "alkoxy" refers to a monovalent radical of the formula RO-, where R is an alkyl as defined herein. Lower alkoxy refers to an alkoxy of 1 -6 carbon atoms, with higher alkoxy is an alkoxy of seven or more carbon atoms. Representative lower alkoxy radicals include methoxy, ethoxy, n-propoxy, n- butoxy, n-pentyloxy, n-hexyloxy, isopropoxy, isobutoxy, isopentyloxy, amyloxy, sec-butoxy, tert-butoxy, tert-pentyloxy, and the like. Higher alkoxy radicals include those corresponding to the higher alkyl radicals set forth herein. The radical may be optionally substituted with substituents at positions that do not significantly interfere with the preparation of compounds falling within the scope of this invention and that do not significantly reduce the efficacy of the compounds. The alkyl is optionally substituted with one to five substituents independently selected from the group consisting of halo, lower akyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino.

[0037] The term "cycloalkyl" refers to a monovalent, alicyclic, saturated hydrocarbon radical having three or more carbons forming the ring. While known cycloalkyl compounds may have up to 30 or more carbon atoms, generally there will be three to seven carbons in the ring. The latter include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. The radical may be optionally substituted with substituents at positions that do not significantly interfere with the preparation of compounds falling within the scope of this invention and that do not significantly reduce the efficacy of the compounds. The cycloalkyl is optionally substituted with one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino.

[0038] The term "hydroxycarbonyl" is a monovolent radical having the formula -C(O)OH.

J0039] The term "lower alkoxycarbonyl" is a monovalent radical having the formula -C(O)OAIk, where AIk is lower alkyl.

[0040] The term "lower alkylcarboxyloxy" is a monovalent radical having the formula -OC(O)AIk, where AIk is lower alkyl.

[0041] The term "lower alkylcarbonylamino" is a monovalent radical having the formula -NHC(O)AIk, where AIk is lower alkyl.

[0042] A "halo" substitutent is a monovalent halogen radical chosen from chloro, bromo, iodo, and fluoro. A "halogenated" compound is one substituted with one or more halo substituent.

[0043] A "1-naphthyl" or "2-naphthyl" is a radical formed by removal of a hydrogen from the 1- or 2-position of a naphthalene structure, respectively. It is optionally substituted with from one to four substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, formyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino.

[0044] A "phenyl" is a radical formed by removal of a hydrogen from a benzene ring. The phenyl is optionally substituted with from one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, carbonyl, hydroxycarbonyl, lower alkylcarbonyloxy, benzyloxy, optionally substituted piperidino, lower alkoxycarbonyl, and lower alkylcarbonylamino.

[0045] A "cyclic amino" is a monovalent radical of a saturated 5-, 6-, or

7-membered cyclic amine ring having no more than one additional hetero atom such as nitrogen, oxygen, or sulfur. Representative examples include, e.g., 1- pyrrolidino, 1 -piperidino, morpholino, piperazino, and the like. These may be substituted or unsubstituted. If substituted, generally they will have no more than 2 substituents chosen from lower alkyl, lower cycloalkyl, hydroxy lower alkyl, phenyl (substituted or unsubstituted), behyzl (substituted or unsubstituted), aminocarbonylmethyl, lower alkylaminocarbonylmethyl, amino, mono- or di-lower alkylamino, or cyclic amino.

[0046] A "carbamoyloxy" is a monovalent radical of the formula

R_t3 R₁₄ NC(O)O- (i.e. an aminocarbonyloxy) where Rn and Rj₄ together form a cyclic amino with the nitrogen atom, or each of R13 and R₁₄ is independently hydrogen, lower alkyl, hydroxy lower alkyl, hydroxy lower alkyl, amino lower alkyl, lower cycloalkyl, phenyl (substituted or unsubstituted), or benzyl (substituted or unsubstituted). Examples include aminocarbonyloxy, methylaminocarbonyloxy, dimethyl aminocarbonyloxy, [4-(l-piperidino)-l- piperidinojcarbonyloxy, l-morpholinocarbonyloxy, 1-pyrrolidinyl, 1- piperazinecarbonyloxy, and others delineated herein.

[0047] A "5-membered heterocyclic ring" is a monovalent radical of a

5-member closed ring containing carbon and at least one other element, generally nitrogen, oxygen, or sulfur and may be fully saturated, partially saturated, or unsaturated (i.e. aromatic in nature). Generally the heterocycle will contain no more than two hetero atoms. Representative examples of unsaturated 5-membered heterocycles with only one hetero atom include 2- or 3-pyrrolyl, 2- or 3-furanyl, and 2- or 3-thiophenyl. Corresponding partially saturated or fully saturated radicals include 3-pyrrolin-2-yl, 2- or 3- pyrrolidinyl, 2- or 3-tetrahydrofuranyl, and 2- or 3-tetrahydrothiophenyl. Representative unsaturated 5-membered heterocyclic radicals having two hetero atoms include imidazolyl, oxazolyl, thiazolyl, pyrazolyl, and the like. The corresponding fully saturated and partially saturated radicals are also included. The heterocyclic radical is bonded through an available carbon atom in the heteocyclic ring. The radical may be optionally substituted with substituents at positions that do not significantly interfere with the preparation of compounds falling within the scope of this invention and that do not significantly reduce the efficacy of the compounds. The ring is optionally substituted with one or two substituents selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino. A "6-membered heterocyclic ring" is a monovalent radical of a

6-member closed ring containing carbon and at least one other element, generally nitrogen, oxygen, or sulfur and may be fully saturated, partially saturated, or unsaturated (i.e. aromatic in nature). Generally the heterocycle will contain no more than two hetero atoms. Representative examples of unsaturated 6-membered heterocycles with only one hetero atom include 2-,

3-, or 4-pyridinyl and 2H-pyranyl. Corresponding partially saturated or fully saturated radicals include 2-, 3-, or 4-piperidinyl, 2-, 3-, or 4-tetrahydropyranyl and the like. Representative unsaturated 6-membered heterocyclic radicals having two hetero atoms include 3- or 4-pyridazinyl, 2-, 4-, or 5-pyrimidinyl,

2-pyrazinyl, and the like. The corresponding fully saturated and partially saturated radicals are also included, e.g. 2-piperazine. The heterocyclic radical is bonded through an available carbon atom in the heterocyclic ring. The radical may be optionally substituted with substituents at positions that do not significantly interfere with the preparation of compounds falling within the scope of this invention and that do not significantly reduce the efficacy of the compounds. The ring is optionally substituted with one or two substituents selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino . [0049] Useful heteroaryl groups include thienyl, benzo[b]thienyl, naphtho[2,3- b]thienyl, thianthrenyl, furyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxanthenyl, 2H-pyrrolyl₅ pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl_J isoquinolyl, quinolyl, phthalzinyl, naphthyridinyl, quinozalinyl, cinnolinyl, pteridinyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl, phenazinyl, isothiazolyl, phenothiazinyl, isoxazolyl, furazanyl, phenoxazinyl, 1,4- dihydroquinoxaline-2,3-dione, 7-aminoisocoumarin, pyrido[l ,2~a]pyrimidin-4- one, l,2-benzoisoxazol-3-yl, benzimidazolyl, 2-oxindolyl, and 2- oxobenzimidazolyl. Where the heteroaryl group contains a nitrogen atom in a ring, such nitrogen atom may be in the form of an N-oxide, e.g., a pyridyl N- oxide, pyrazinyl N-oxide, pyrimidinyl N-oxide, and the like.

[0050] A "fused 2-, 3-, or 4-ring heterocyclic radical" is polynuclear in that the adjacent rings share a pair of atoms, generally carbon atoms. At least one of the rings will be heterocyclic in that it will have a noncarbon atom such as nitrogen, oxygen, or sulfur. The ring system may contain from 9 to 18 atoms. A 2-ring heterocyclic system will generally have 9 or 10 atoms included in the ring. Examples of such a 2-ring system include quinoline, isoquinoline, purine, indolizine, 4H-quinolizine, 3H-pyrrolizine, coumaran, coumarin, isocoumarin, 4-methylcoumarin, 3 -chloro-H-methyl coumarin, chromone, benzofuran, benzothiophene, benzothiazole, indole, and the like. A 3-ring system will generally have 12 to 14 atoms included in the ring. Examples of such a 3-ring system include carbazole, acridine, and the like. A 4-ring fused system will generally have 16 to 18 atoms included in the chain. Examples of such a 4- ring system include isothebaine and the like. The ring is bonded through a ^• carbon in the ring system. The radical may be optionally substituted with substituents at positions that do not significantly interfere with the preparation of compounds falling within the scope of this invention and that do not significantly reduce the efficacy of the compounds. The radical is optionally substituted with one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino,- halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkyl carbonyloxy, and lower alkylcarbonylamino.

[0051] The term "pharmaceutical composition" as used herein, is to be understood as defining compositions of which the individual components or ingredients are themselves pharmaceutically acceptable, e.g., where oral administration is foreseen, acceptable for oral use; where topical administration is foreseen, topically acceptable; and where intravenous administration is foreseen, intravenously acceptable.

[0052] As used herein, the term "therapeutically effective amount" refers to that amount of the therapeutic agent sufficient to result in amelioration of one or more symptoms of a disorder, or prevent advancement of a disorder, or cause regression of the disorder. For example, with respect to the treatment of cancer, a therapeutically effective amount preferably refers to the amount of a therapeutic agent that decreases the rate of tumor growth, decreases tumor mass, decreases the number of metastases, increases time to tumor progression, or increases survival time by at least 5%, preferably at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%.

[0053] The terms "prevent," "preventing," and "prevention," as used herein, refer to a decrease in the occurrence of pathological cells (e.g., hyperproliferative or neoplastic cells) in an animal. The prevention may be complete, e.g., the total absence of pathological cells in a subject. The prevention may also be partial, such that the occurrence of pathological cells in a subject is less than that which would have occurred without the present invention.

[0054] Compounds having Formula I can be provided as pharmaceutically acceptable salts. Examples of pharmaceutically acceptable salts (i.e., addition salts) include inorganic and organic acid addition salts such as hydrochloride, hydrobromide, phosphate, sulphate, citrate, lactate, tartrate, maleate, fumarate, mandelate, benzoate and oxalate; and inorganic and organic base addition salts with bases such as sodium hydroxyide, Tris(hydroxymethyl)aminomethane (TRIS, tromethane) and N-methyl-glucamine. Although the salts typically have similar physiological properties compared to the free base, certain acid addition salts may demonstrate preferred physicochemical properties, e.g., enhanced solubility, improved stability. One particular pharmaceutically acceptable salt is derived from maleic acid, the salt being either a hydrogen maleate or a dimaleate salt.

{0055] Certain of the compounds of the present invention may exist as stereoisomers including optical isomers. The invention includes all stereoisomers and both the racemic mixtures of such stereoisomers as well as the individual enantiomers that may be separated according to methods that are well known to those of ordinary skill in the art. For example, camptothecin analogs of the present invention may have 20(R), 20(S) or 20(RS) configuration (i.e., a racemic mixture). Similarly, camptothecin analog Nj-oxides and bis N-oxides of the present invention may have 20(R), 20(S) or 20(RS) configuration (i.e., a racemic mixture). Certain of the compounds of the present invention may also exist as diasteroisomers wherein one or more substituents on the camptothecin analog contain one or more chiral centers.

[0056] In certain embodiments of the invention, compounds having Formula I are administered in combination with one or more other active agents (e.g., chemotherapeutic agents) or treatments. By way of non-limiting example, a patient may be treated for a hyperproliferative disorder, such as cancer, by the administration of a therapeutically effective amount of a compound having Formula I in combination with radiotherapy agent/treatment or the administration of a chemotherapeutic agent.

[0057] In other embodiments, compounds of the invention are administered in combination with agents, such as anti -angiogenic agents, that block inhibit or modulate tumor neovascularization. In preferred embodiments, anti- angiogenesis agents can be any anti-angiogenesis agent which is used, has been used, or is known to be useful for the treatment of hyperproliferative disorders. Examples of anti-angiogenesis agents include bevacizumab (Avastin™), VEGF-TRAP, anti-VEGF-receptor antibodies, angiostatin, endostatin, batimastat, captopril, cartilage derived inhibitor, genistein, interleukin 12, lavendustin, medroxypregesterone acetate, recombinant human platelet factor 4, tecogalan, thrombospondin, TNP-470, VEGF antagonists, anti-VEGF monoclonal antibody, soluble VEGF-receptor chimaeric protein, antisense oligonucleotides, antisense oligodexoynucleotides, siRNAs, anti- VEGF aptamers, pigment epithelium derived factor, a tyrosine kinase inhibitor, an inhibitor of epidermal-derived growth factor, an inhibitor of fibroblast-derived growth factor, an inhibitor of platelet derived growth factor, an MMP (matrix metalloprotease) inhibitor, an integrin blocker, interferon-α, pentosan polysulfate, a cyclooxygenase inhibitor, carboxyamidotriazole, combretastatin A-4, squalamine, 6-O-chloroacetyl-carbonyl)-fumagillol, thalidomide, troponin- 1, indolinethiones, pyridopyrimidines, quinoazolines, phenyl-pyrrolo-pyrimidines, trastuzumab, calcium influx inhibitor (CAI), neomycin, squalamine, marimastat, prinomastat (AG-3340), metastat (COL-3) and cinnoline derivatives. Additional an ti -angiogenic compounds that may be administered in combination with the compounds of the present invention are described in U.S. Patent Nos. 5,192,744, 5,426,100, 5,733,876, 5,840,692, 5,854,205, 5,990,280, 5,994,292, 6,342,219, 6,342,221, 6,346,510, 6,479,512, 6,719,540, 6,797,488, 6,849,599, 6,869,952, 6,887,874, 6,958,340 and 6,979,682. In certain embodiments, the compounds of the present invention are administered in combination with a vascular targeting agent (also known as vascular damaging agents). In one embodiment, the vascular targeting agent is for the treatment of malignant or non-malignant vascular proliferative disorders. In other embodiments, vascular targeting agents can be any vascular targeting agent which is used, has been used, or is known to be useful for the treatment of hyperproliferative disorders. Examples of vascular targeting agents that may be administered in combination with the compounds of the present invention include DMXAA 5,6-dimethylxanthenone-4-acetic acid, ZD6126, (5S)-5-(acetylamino)-9,10,l l-tτimethoxy-6,7-dihydro-5i7- dibenzo[a,c]cyclohepten-3-yl dihydrogen phosphate, also known as JV-acetylcolchinol-O-phosphate (see, for example, U.S. Patent No. 6,906,048); functionalized stilbene derivatives such as combretastatin A4 and its prodrugs (see, e.g., U.S. Patent Nos. 6,919,324 and 6,773,702); dioleoyltrimethyl- ammonium propane (DOTAP), N-[l-(2,3-dioleoyloxy)-propyl]-N,N,N- trimethylammonium chloride (DOTMA), dimethyldioctadecylammonium bromide (DDAB), l,2-dimyristyloxypropyl-3-dimethylhydroxyethyl (DMRJE), dioleoyl-3-dimethylammonium propane (DODAP), N,N-dioleyl-N,N- dimethylammonium chloride (DODAC), or N-(l-(2,3-dioleyloxy)propyl)-N- (2-(sperminecarboxamido)ethyl)-N,N-dimethyl ammonium trifluoroacetate (DOSPA), or any other natural or synthetic cationic lipids, including, for example, dioleoylphosphatidyl-choline (DOPC), dipalmitoylphosphatidylcholine (DPPC), disteroylphosphatidylcholine (DSPC), dimyristoylphosphatidylcholine (DMPC), or 1,2-sn- dioleoylphosphatidylcholine (DOPE), or any other natural or synthetic electrostatically neutral lipids (see, for example, U.S. patent No. 6,680,068); vascular targeting agents which incorporate benzo[6]thiophene, indole, and benzofuran molecular skeletons such as those described in U.S. Patent No. 6,593,374. In other embodiments, the compounds of the present invention are administered in combination with a hypoxia-inducible factor 1 (HIFl) inhibitor. In one embodiment, the HEFl inhibitor is for the treatment of malignant or non-malignant vascular proliferative disorders. In other embodiments, HEFl inhibitors can be any HIFl inhibitor which is used, has been used, or is known to be useful for the treatment of hyperproliferative disorders. Examples of HIFl inhibitors suitable for use in combination with compounds of the present invention include topotecan, Pl 3 kinase inhibitors; LY294002; rapamycin; histone deacetylase inhibitors such as [(E)- (lS,4S,10S,21R)-7-[(Z)-ethylidene]-4,21 -diisopropyl-2-oxa-12,13-dithia- 5,8,20,23-tetraazabicyclo-[8,7,6]-tricos-16-ene-3,6,9,19,22-pentanone (FR901228, depsipeptide); heat shock protein 90 (Hsp90) inhibitors such as geldanamycin, 17-allylamino-geldanamycin (17-AAG), and other geldanamycin analogs, and radicicol and radicicol derivatives such as KF58333; genistein; indanone; staurosporin; protein kinase-1 (MEK-I) inhibitors such as PD98059 (2'-amino-3'-methoxyflavone); PX-12 (1- methylpropyl 2-imidazolyl disulfide); pleurotin PX478; quinoxaline 1,4- dioxides; sodium butyrate (NaB); sodium nitropurruside (SNP) and other NO donors; microtubule inhibitors such as novobiocin, panzem (2- melhoxyestradiol or 2-ME2), vincristines, taxanes, epothilones, discodermolide, and derivatives of any of the foregoing; coumarins; barbituric and thiobarbituric acid analogs; camptothecins; and YC-I. See U.S. Patent No. 6,979,675.

[0060] In certain embodiments, the compounds of the present invention are administered in combination with an Hsp90 inhibitor. In one embodiment, the Hsp90 inhibitor is for the treatment of malignant or non-malignant vascular proliferative disorders. In other embodiments, Hsp90 inhibitors can be any Hsp90 inhibitor which is used, has been used, or is known to be useful for the treatment of hyperproliferative disorders. Examples of Hsp90 inhibitors that may be combined with the compounds of the present invention include geldanamycin, 17-allylamino-17-demethoxygeldanamycin, geldanamycin derivatives such as those described in U.S. patent No. 6,890,917, dexamethasone and benzoquinone ansamycins such as those described in U.S. patent No. 6,872,715. Additional Hsp90 inhibitors are disclosed in U.S. Patent Nos. 6,613,780, 6,281,229 and 6,903,116.

[0061] In other embodiments, the compounds of the present invention are administered in combination with an inhibitor of tyrosine and/or serine/threonine kinases and tyrosine kinase receptors involved in cellular signaling. These include tyrosine kinase inhibitors of Src, AbI, Platelet Derived Growth Factor Receptors, Vascular Endothelial Growth Factor Receptors, c-Met, Fibroblast Growth Factor receptors, Epidermal Growth Factor Receptors, Insulin Growth Factor Receptors, mTOR, Flt-3, CSF-I Receptor, AKT, Polo kinases, Aurora Kinases, STAT-3, PI-3 Kinase, Ras, Raf and Mitogen Activated Kinases, MEK, ERK. Examples of tyrosine kinase and serine/threonine kinase inhibitors include (but not limited to): AMG706, ZA6474, BAY 43-9006, Dasatinib, CEP-701, XL647, XL999. Lapatinb, MLN5187CT53518, PKC412, ST1571, AMN107, AEE 788, OSI-930, OSI- 817, SU11248, AG-03736, GW-786034m , CEP-7055.

[0062] In other embodiments, the compounds of the present invention are administered in combination with HDAC inhibitors. Examples include (but noflimited to) SAHA, MS-275, MGCD0103, LBH589, PXDlOl, FK228.

[0063] In other embodiments, the compounds of the present invention are administered in combination with proteasome inhbitors such as Velcade.

[0064] In other embodiments, the compounds of the present invention are administered in combination with pro-apoptotic agents such as TRAIL, anti- DR4/DR5 (TRA8) antibodies, IAP, Survivin or small molecules that stimulate caspase activation.

[0065] In other embodiments, the compounds of the present invention are administered in combination with inhibitors of cell cycle regulators such as CDK inhibitors.

[0066] "In combination" refers to the use of more than one treatment. The use of the term "in combination" does not restrict the order in which treatments are administered to a subject being treated for a hyperproliferative disorder. A first treatment can be administered prior to, concurrently with, after, or within any cycling regimen involving the administration of a second treatment to a subject with a hyperproliferative disorder. For example, the first treatment can be administered .5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before a treatment; or the first treatment can be administered 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after a second treatment. Such treatments include, for example, the administration of compounds having Formula I in combination with one or more chemotherapeutic agents or radiotherapeutic agents/treatments.

[0067] The term "chemotherapeutic agent," as used herein, is intended to refer to any chemotherapeutic agent known to those of skill in the art to be effective for the treatment, prevention or amelioration of hyperproliferative disorders such as cancer. Chemotherapeutic agents include, but are not limited to, small molecules, synthetic drugs, peptides, polypeptides, proteins, nucleic acids {e.g., DNA and RNA polynucleotides including, but not limited to, anti sense nucleotide sequences, triple helices and nucleotide sequences encoding biologically active proteins, polypeptides or peptides), antibodies, synthetic or natural inorganic molecules, mimetic agents, and synthetic or natural organic molecules. Any agent which is known to be useful, or which has been used or is currently being used for the treatment or amelioration of a hyperproliferative disorder can be used in combination with a compound having Formula I. See, e.g., Hardman et ah, eds., 2002, Goodman & Gilman's The Pharmacological Basis Of Therapeutics 10th Ed, Mc-Graw-Hill, New York, NY for information regarding therapeutic agents which have been or are currently being used for the treatment or amelioration of a hyperproliferative disorder.

[0068] Particular chemotherapeutic agents useful in the methods and compositions of the invention include alkylating agents, antimetabolites, antimitotic agents, epipodophyllotoxins, antibiotics, hormones and hormone antagonists, enzymes, platinum coordination complexes, anthracenediones, substituted ureas, methylhydrazine derivatives, imidazotetrazine derivatives, cytoprotective agents, DNA topoisomerase inhibitors, biological response modifiers, retinoids, therapeutic antibodies, differentiating agents, immunomodulatory agents, angiogenesis inhibitors and anti-angiogenic agents.

[0069] Certain chemotherapeutic agents include, but are not limited to, abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic trioxide, asparaginase, BCG live, bevaceizumab, bexarotene, bleomycin, bortezomib, busulfan, calusterone, camptothecin, capecitabine, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cinacalcet, cisplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, darbepoetin alfa, daunorubicin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone, Elliott's B solution, epirubicin, epoetin alfa, estramustine, etoposide, exemestane, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gemcitabine, gemtuzumab ozogamicin, gefitinib, goserelin, hydroxyurea, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib, interferon alfa-2a, interferon alfa-2b, irinotecan, letrozole, leucovorin, levamisole, lomustine, meclorethamine, megestrol, melphalan, mercaptopurine, mesna, methotrexate, methoxsalen, methylprednisolone, mitomycin C, mitotane, mitoxantrone, nandroione, nofetumomab, oblimersen, oprelvekin, oxaliplatin, paclitaxel, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed, pentostatin, pipobroman, plicamycin, polifeprosan, porfimer, procarbazine, quinacrine, rasburicase, rituximab, sargramostim, streptozocin, talc, tamoxifen, tarceva, temozolomide, teniposide, testolactone, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, and zoledronate. In certain embodiments, chemotherapeutic agents are selected from gemcitabine and irinotecan.

[0070] Chemotherapeutic agents may be administered at doses that are recognized by those of skill in the art to be effective for the treatment of the hyperproliferative disorder. In certain embodiments, chemotherapeutic agents may be administered at doses lower than those used in the art due to the additive or synergistic effect of the compounds having Formula I.

[0071] Therapeutic agents useful in the methods and compositions of the invention include active vitamin D compound or mimics thereof, antineoplastic agents (e.g., actinomycin D, irinotecan, vincristine, vinorelbine, SN-38, azacitidine (5-azacytidine, 5AzaC), thalidomide vinblastine, methotrexate, azathioprine, fluorouracil, doxorubicin, mitomycin, docetaxel. paclitaxel), angiogenic inhibitors (e.g., VEGF-TRAP, angiostatin, endostatin, aptamer antogonist of VEGF, batimastat, captopril, cartilage derived inhibitor, genistein, interleukin 12, lavendustin, medroxypregesterone acetate, ^• recombinant human platelet factor 4, tecogalan, thrombospondin and TNP-470), serine/threonine kinase inhibitors, tyrosine kinase inhibitors, HDAC inhibitors, Proteasome inhibitors, CDK inhibitors, HSP inhibitors, vasodilators (e.g., nitrates, calcium channel blockers), anticoagulants {e.g., heparin), anti-platelet agents (e.g., aspirin, blockers of Eb/IIIa receptors, clopidogrel), anti-thrombins (e.g., hirudin, iloprost), immunosuppressants (e.g., sirolimus, tranilast, dexamethasone, tacrolimus, everolimus, A24), collagen synthetase inhibitors (e.g., halofuginone, propyl hydroxylase, C- proteinase inhibitor, metalloproteinase inhibitor), antiinflammatories (e.g., corticosteroids, non-steroidal anti-inflammatory drugs), 17β-estradio], angiotensin converting enzyme inhibitors, colchicine, fibroblast growth factor antagonists, histamine antagonists, lovastatin, nitroprusside, phosphodiesterase inhibitors, prostaglandin inhibitors, suramin, serotonin blockers, thioprotease inhibitors, platelet-derived growth factor antagonists, nitric oxide, and angiopeptin. hi one embodiment, the therapeutic agent is a taxane, e.g., paclitaxel or docetaxel.

[0072] In certain embodiments, patients are subjected to a hypoxia imaging technique prior to administration of the compositions comprising the compounds of the present invention. Examples of imaging techniques suitable for the determination of the presence of hypoxic tumor cells include computed tomography (CT), magnetic resonance imaging (MRI), single photon emission computer tomography (SPECT), and positron emission tomography (PET). Use of such visualization methods can advantageously be used to select a subset of patients that are particularly suitable for treatment with hypoxia activated antiproliferative compositions of the present invention.

[0073] In this embodiment, the invention is directed to a method of treating, preventing or ameliorating a hyperproliferative disease in an animal in need thereof, comprising determining whether said hyperproliferative disease is characterized by hypoxic tissue, and treating said animal with an effective amount of a compound of the invention.

[0074] The term "radiotherapeutic agent," as used herein, is intended to refer to any radiotherapeutic agent known to one of skill in the art to be effective to treat or ameliorate cancer, without limitation. For instance, the radiotherapeutic agent can be an agent such as those administered in brachytherapy or radionuclide therapy. Such methods can optionally further comprise the administration of one or more additional cancer therapies, such as, but not limited to, chemotherapies, surgery, and/or another radiotherapy.

[0075] In certain embodiments involving radiotherapeutic agents or treatments, the present invention relates to a method for treating cancer comprising the administration of camptothecin or analog N-oxide having Formula I, in combination with a treatment comprising a therapeutically effective dose of brachytherapy. The brachytherapy can be administered according to any schedule, dose, or method known to one of skill in the art to be effective in the treatment or amelioration of cancer, without limitation. In general, brachytherapy comprises insertion of radioactive sources into the body of a subject to be treated for cancer, preferably inside the tumor itself, such that the tumor is maximally exposed to the radioactive source, while preferably minimizing the exposure of healthy tissue.

[0076] In certain embodiments, the brachytherapy can be intracavitary brachytherapy. In other embodiments, the brachytherapy can be interstitial brachytherapy. Whether the brachytherapy is intracavitary brachytherapy or interstitial brachytherapy, the brachytherapy can be administered at a high dose rate, a continuous low dose rate, or a pulsed dose rate. For example, and not by way of limitation, a high dose rate brachytherapy regimen can be a dose of 60 Gy administered in ten fractions over six days, while a continuous low dose rate brachytherapy regimen can be a total dose of about 65 Gy, administered continuously at about 40 to 50 cGy per hour. Other examples of high, continuous low, and pulsed dose rate brachytherapy are well known in the art. See, e.g., Mazeron et al, Sent. Rad. One. 12:95-108 (2002). [0077] Representative radioisotopes that can be administered in any of the above-described brachytherapies include, but are not limited to, phosphorus 32, cobalt 60, palladium 103, ruthenium 106, iodine 125, cesium 137, indium 192, xenon 133, radium 226, californium 252, or gold 198. Other radioisotopes may be selected for administration in brachytherapy according to the desirable physical properties of such a radioisotope. One of skill in the art will readily recognize that many properties will affect a radioisotope's suitability for use in brachytherapy, including, but not limited to, the radioisotope's half-life, the degree to which emitted radiation penetrates surrounding tissue, the energy of emitted radiation, the ease or difficulty of adequately shielding the radioisotope, the availability of the radioisotope, and the ease or difficulty of altering the shape of the radioisotope prior to administration.

[0078] Additional methods of administering and apparatuses and compositions useful for brachytherapy are described in U.S. Patent Nos. 6,319,189, 6,179,766, 6,168,777, 6,149,889, and 5,611,767, each of which is incorporated herein by reference in its entirety.

[0079] In certain embodiments involving radiotherapeutic agents or treatments, the present invention relates to a method for treating cancer comprising the administration of camptothecin or analog N-oxide having Formula I, in combination with a treatment comprising a therapeutically effective dose of a radionuclide. The radionuclide therapy can be administered according to any schedule, dose, or method known to one of skill in the art to be effective in the treatment or amelioration of cancer, without limitation. In general, radionuclide therapy comprises systemic administration of a radioisotope that preferentially accumulates in or binds to the surface of cancerous cells. The preferential accumulation of the radionuclide can be mediated by a number of mechanisms, including, but not limited to, incorporation of the radionuclide into rapidly proliferating cells, specific accumulation of the radionuclide by the cancerous tissue without special targeting (e.g., iodine 131 accumulation in thyroid cancer), or conjugation of the radionuclide to a biomolecule specific for a neoplasm.

[0080J Representative radioisotopes that can be administered in radionuclide therapy include, but are not limited to, phosphorus 32, yttrium 90, dysprosium 165, indium 111, strontium 89, samarium 153, rhenium 186, iodine 131, iodine 125, lutetium 177, and bismuth 213. While all of these radioisotopes may be linked to a biomolecule providing specificity of targeting, iodine 131, indium 111, phosphorus 32, samarium 153, and rhenium 186 may be administered systemically without such conjugation. One of skill in the art may select a specific biomolecule for use in targeting a particular neoplasm for radionuclide therapy based upon the cell-surface molecules present on that neoplasm. For example, hepatomas may be specifically targeted by an antibody specific for ferritin, which is frequently over-expressed in such tumors. Examples of antibody-targeted radioisotopes for the treatment of cancer include ZEV ALEST (ibritumomab tiuxetan) and BEXXAR (tositumomab), both of which comprise an antibody specific for the B cell antigen CD20 and are used for the treatment of non-Hodgkin lymphoma.

[0081 J Other examples of biomolecules providing specificity for particular cell are reviewed in an article by Thomas, Cancer Biother. Radiopharm. J 7:71-82 (2002), which is incorporated herein by reference in its entirety. Furthermore, methods of administering and compositions useful for radionuclide therapy may be found in U.S. Patent Nos. 6,426,400, 6,358,194, 5,766,571, and 5,563,250, each of which is incorporated herein by reference in its entirety.

[0082] In certain embodiments involving radiotherapeutic agents or treatments, the present invention relates to a method for treating cancer comprising the administration of camptothecin or analog N-oxide having Formula I, in combination with a treatment comprising a therapeutically effective dose of external-beam radiation therapy. The external-beam radiation therapy can be administered according to any schedule, dose, or method known to one of skill in the art to be effective in the treatment or amelioration of cancer, without limitation. In. general, external-beam radiation therapy comprises irradiating a defined volume within a subject with a high energy beam, thereby causing cell death within that volume. The irradiated volume preferably contains the entire cancer to be treated, and preferably contains as little healthy tissue as possible.

[0083] In certain embodiments, the external-beam radiation therapy can be three-dimensional conformal radiotherapy. In other embodiments, the external-beam radiation therapy can be continuous hyperfractionated radiotherapy. In still other embodiments, the external-beam radiation therapy can be intensity-modulated radiotherapy. In yet other embodiments, the external-beam radiation therapy can be helical tomotherapy. In still other embodiments, the external-beam radiation therapy can be three-dimensional conformal radiotherapy with dose escalation. In yet other embodiments, the external-beam radiation therapy can be stereotactic radiotherapy, including, but not limited to, single fraction stereotactic radiotherapy, fractionated stereotactic radiotherapy, and fractionated stereo tactically guided conformal radiotherapy.

[0084] The external-beam radiation therapy can be generated or manipulated by any means known to one of skill in the art. For example, the photon beam used in external-beam radiation therapy can be shaped by a multileaf collimator. Other examples of suitable devices for generating a photon beam for use in external-beam radiation therapy include a gamma knife and a linac- based stereotactic apparatus. In certain embodiments, administration of the external-beam radiation therapy is controlled by a computer according to a three-dimensional model of the patient in the treatment position. Such a model can be generated, for example, by computed tomography (CT), magnetic resonance imaging (MRI), single photon emission computer tomography (SPECT), and positron emission tomography (PET). Use of such visualization methods can advantageously minimize the volume of healthy tissue treated, thereby allowing higher total doses of radiation to be administered to the patient. [0085] In addition, healthy tissues can optionally be protected from the effects of the external-beam radiation therapy by placing blocking devices such as, e.g., lead shields, in locations where such protection is needed. Alternatively or additionally, metal reflecting shields can optionally be located to reflect the photon beam in order to concentrate the radiation on the cancerous tissue to be treated and protect healthy tissue. Placement of either shield is well within the knowledge of one of skill in the art.

[0086] Methods of administering and apparatuses and compositions useful for external-beam radiation therapy can be found in U.S. Patent Nos. 6,449,336, 6,398,710, 6,393,096, 6,335,961, 6,307,914, 6,256,591, 6,245,005, 6,038,283, 6,001,054, 5,802,136, 5,596,619, and 5,528,652, each of which is incorporated herein by reference in its entirety.

[0087] In certain embodiments involving radiotherapeutic agents or treatments, the present invention relates to a method for treating cancer comprising the administration of camptothecin or analog N-oxide having Formula I, in combination with a treatment comprising a therapeutically effective dose of thermotherapy. The thermotherapy can be administered according to any schedule, dose, or method known to one of skill in the art to be effective in the treatment or amelioration of cancer, without limitation. In certain embodiments, the thermotherapy can be cryoablation therapy. In other embodiments, the thermotherapy can be hyperthermic therapy. In still other embodiments, the thermotherapy can be a therapy that elevates the temperature of the tumor higher than in hyperthermic therapy.

[0088] Cryoablation therapy involves freezing of a neoplastic mass, leading to deposition of intra- and extra-cellular ice crystals; disruption of cellular membranes, proteins, and organelles; and induction of a hyperosmotic environment, thereby causing cell death. Cryoablation can be performed in one, two, or more freeze-thaw cycles, and further the periods of freezing and thawing can be adjusted for maximum tumor cell death by one of skill in the art. One exemplary device that can be used in cryoablation is a cryoprobe incorporating vacuum-insulated liquid nitrogen. See, e.g., Murphy et al., Sent. Urol. Oncol. 79:133-140 (2001). However, any device that can achieve a local temperature of about -180⁰C to about -195⁰C can be used in cryoablation therapy. Methods for and apparatuses useful in cryoablation therapy are described in U.S. Patent Nos. 6,383,181, 6,383,180, 5,993,444, 5,654,279, 5,437,673, and 5,147,355, each of which is incorporated herein by reference in its entirety.

[0089] Hyperthermic therapy typically involves elevating the temperature of a neoplastic mass to a range from about 42°C to about 44⁰C. The temperature of the cancer may be further elevated above this range; however, such temperatures can increase injury to surrounding healthy tissue while not causing increased cell death within the tumor to be treated. The tumor may be heated in hyperthermic therapy by any means known to one of skill in the art without limitation. For example, and not by way of limitation, the tumor may be heated by microwaves, high intensity focused ultrasound, ferromagnetic thermoseeds, localized current fields, infrared radiation, wet or dry radiofrequency ablation, laser photocoagulation, laser interstitial thermic therapy, and electrocautery. Microwaves and radiowaves can be generated by waveguide applicators, horn, spiral, current sheet, and compact applicators.

[0090] Other methods of and apparatuses and compositions for raising the temperature of a tumor are reviewed in an article by Wust et at, Lancet Oncol. 3:487-97 (2002), and described in U.S. Patent Nos. 6,470,217, 6,379,347, 6,165,440, 6,163,726, 6,099,554, 6,009,351, 5,776,175, 5,707,401, 5,658,234, 5,620,479, 5,549,639, and 5,523,058, each of which is incorporated herein by reference in its entirety.

[0091] In certain embodiments involving radiotherapeutic agents or treatments, the present invention relates to a method for treating cancer comprising the administration of camptothecin or analog N-oxide having Formula I, in combination with a treatment comprising a therapeutically effective dose of radiosurgery. The radiosurgery can be administered according to any schedule, dose, or method known to one of skill in the art to be effective in the treatment or amelioration of cancer, without limitation. In general, radiosurgery comprises exposing a defined volume within a subject to a manually directed radioactive source, thereby causing cell death within that volume. The irradiated volume preferably contains the entire cancer to be treated, and preferably contains as little healthy tissue as possible. Typically, the tissue to be treated is first exposed using conventional surgical techniques, then the radioactive source is manually directed to that area by a surgeon. Alternatively, the radioactive source can be placed near the tissue to be irradiated using, for example, a laparoscope. Methods and apparatuses useful for radiosurgery are further described in Valentini et al., Eur. J. Surg. Oncol. 28:180-185 (2002) and in U.S. Patent Nos. 6,421,416, 6,248,056, and 5,547,454, each of which is incorporated herein by reference in its entirety.

[0092] In certain embodiments involving radiotherapeutic agents or treatments, the present invention relates to a method for treating cancer comprising the administration of camptothecin or analog N-oxide having Formula I, in combination with a treatment comprising a therapeutically effective dose of charged-particle radiotherapy. The charged-particle radiotherapy can be administered according to any schedule, dose, or method known to one of skill in the art to be effective in the treatment or amelioration of cancer, without limitation. In certain embodiments, the charged-particle radiotherapy can be proton beam radiotherapy. In other embodiments, the charged-particle radiotherapy can be helium ion radiotherapy. In general, charged-particle radiotherapy comprises irradiating a defined volume within a subject with a charged-particle beam, thereby causing cellular death within that volume. The irradiated volume preferably contains the entire cancer to be treated, and preferably contains as little healthy tissue as possible. A method for administering charged-particle radiotherapy is described in U.S. Patent No. 5,668,371, which is incorporated herein by reference in its entirety.

[0093] In certain embodiments involving radiotherapeutic agents or treatments, the present invention relates to a method for treating cancer comprising the administration of camptothecin or analog N-oxide having Formula I, in combination with a treatment comprising a therapeutically effective dose of neutron radiotherapy. The neutron radiotherapy can be administered according to any schedule, dose, or method known to one of skill in the art to be effective in the treatment or amelioration of cancer, without limitation.

[0094] In certain embodiments, the neutron radiotherapy can be a neutron . capture therapy. In such embodiments, a compound that emits radiation when bombarded with neutrons and preferentially accumulates in a neoplastic mass is administered to a subject. Subsequently, the tumor is irradiated with a low energy neutron beam, activating the compound and causing it to emit decay products that kill the cancerous cells. Such compounds are typically boron containing compounds, but any compound that has a significantly larger neutron capture cross-section than common body constituents can be used. The neutrons administered in such therapies are typically relatively low energy neutrons having energies at or below about 0.5 eV. The compound to be activated can be caused to preferentially accumulate in the target tissue according to any of the methods useful for targeting of radionuclides, as described below, or in the methods described in Laramore, Semin. Oncol. 24:672-685 (1997) and in U.S. Patents Nos. 6,400,796, 5,877,165, 5,872,107, and 5,653,957, each of which is incorporated herein by reference in its entirety.

[0095] In other embodiments, the neutron radiotherapy can be a fast neutron radiotherapy. In general, fast neutron radiotherapy comprises irradiating a defined volume within a subject with a neutron beam, thereby causing cellular death within that volume. The irradiated volume preferably contains the entire cancer to be treated, and preferably contains as little healthy tissue as possible. Generally, high energy neutrons are administered in such therapies, with energies in the range of about 10 to about 100 million eV. Optionally, fast neutron radiotherapy can be combined with charged-particle radiotherapy in the administration of mixed proton-neutron radiotherapy.

[0096] In certain embodiments involving radiotherapeutic agents or treatments, the present invention relates to a method for treating cancer comprising the administration of camptothecin or analog N-oxide having Formula I, in combination with a treatment comprising a therapeutically effective dose of photodynamic therapy. The photodynamic therapy can be administered according to any schedule, dose, or method known to one of skill in the art to be effective in the treatment or amelioration of cancer, without limitation. In general, photodynamic therapy comprises administering a photosensitizing agent that preferentially accumulates in a neoplastic mass and sensitizes the neoplasm to light, then exposing the tumor to light of an appropriate wavelength. Upon such exposure, the photosensitizing agent catalyzes the production of a cytotoxic agent, such as, e.g., singlet oxygen, which kills the cancerous cells.

[0097] Representative photosensitizing agents that may be used in photodynamic therapy include, but are not limited to, porphyrins such as porfimer sodium, 5-aminolaevulanic acid and verteporfin; chlorins such as temoporfin; texaphyrins such as lutetium texephyrin; purpurins such as tin etiopurpurin; phthalocyanines; and titanium dioxide. The wavelength of light used to activate the photosensitizing agent can be selected according to several factors, including the depth of the tumor beneath the skin and the absorption spectrum of the photosensitizing agent administered. The period of light exposure may also vary according to the efficiency of the absorption of light by the photosensitizing agent and the efficiency of the transfer of energy to the cytotoxic agent. Such determinations are well within the ordinary skill of one in the art.

[0098] Methods of administering and apparatuses and compositions useful for photodynamic therapy are disclosed in Hopper, Lancet Oncol. 7:212-219 (2000) and U.S. Patent Nos. 6,283,957, 6,071,908, 6,011,563, 5,855,595, 5,716,595, and 5,707,401, each of which is incorporated herein by reference in its entirety.

[0099] It will be appreciated that both the particular radiation dose to be utilized in treating a hyperproliferative disorder and the method of administration will depend on a variety of factors. Thus, the dosages of radiation that can be used according to the methods of the present invention are determined by the particular requirements of each situation. The dosage will depend on such factors as the size of the tumor, the location of the tumor, the age and sex of the patient, the frequency of the dosage, the presence of other tumors, possible metastases and the like. Those skilled in the art of radiotherapy can readily ascertain the dosage and the method of administration for any particular tumor by reference to Hall, E. J., Radiobiology for the Radiologist, 5th edition, Lippincott Williams & Wilkins Publishers, Philadelphia, PA, 2000; Gunderson, L. L. and Tepper J. E., eds., Clinical Radiation Oncology, Churchill Livingstone, London, England, 2000; and Grosch, D. S., Biological Effects of Radiation, 2nd edition, Academic Press, San Francisco, CA, 1980. In certain embodiments, radiotherapeutic agents and treatments may be administered at doses lower than those known in the art due to the additive or synergistic effect of the compound having Formula 1. ] . Compositions in accordance with the present invention may be employed for administration in any appropriate manner, e.g., oral or buccal administration, e.g., in unit dosage form, for example in the form of a tablet, in a solution, in hard or soft encapsulated form including gelatin encapsulated form, sachet, or lozenge. Compositions may also be administered parenterally or topically, e.g., for application to the skin, for example in the form of a cream, paste, lotion, gel, ointment, poultice, cataplasm, plaster, dermal patch or the like, or for ophthalmic application, for example in the form of an eyedrop, -lotion or -gel^' formulation. Readily flowable forms, for example solutions, emulsions and suspensions, may also be employed e.g., for intralesional injection, or may be administered rectally, e.g., as an enema or suppository, or intranasal administration, e.g., as a nasal spray or aerosol. Microcrystalline powders may be formulated for inhalation, e.g. , delivery to the nose, sinus, throat or lungs. Transdermal compositions/devices and pessaries may also be employed for delivery of the compounds of the invention. The compositions may additionally contain agents that enhance the delivery of the compounds having Formula I (or other active agents), e.g., liposomes, polymers or co-polymers (e.g., branched chain polymers). Preferred dosage forms of the present invention include oral dosage" forms and intravenous dosage forms.

[00101] Intravenous forms include, but are not limited to, bolus and drip injections. In preferred embodiments, the intravenous dosage forms are sterile or capable of being sterilized prior to administration to a subject since they typically bypass the subject's natural defenses against contaminants. Examples of intravenous dosage forms include, but are not limited to, Water for Injection USP; aqueous vehicles including, but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water-miscible vehicles including, but not limited to, ethyl alcohol, polyethylene glycol and polypropylene glycol; and non-aqueous vehicles including, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate and benzyl benzoate.

[00102] The pharmaceutical compositions of the present invention may further comprise one or more additives. Additives that are well known in the art include, e.g., detackifiers, anti-foaming agents, buffering agents, antioxidants (e.g., ascorbic acid, ascorbyl palmitate, sodium ascorbate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate, malic acid, fumaric acid, potassium metabisulfite, sodium bisulfite, sodium metabisulfite, and tocopherols, e.g., α-tocopherol (vitamin E)), preservatives, chelating agents, viscomodulators, tonicifiers, flavorants, colorants, odorants, opacifϊers, suspending agents, binders, fillers, plasticizers, lubricants, and mixtures thereof. The amounts of such additives can be readily determined by one skilled in the art, according to the particular properties desired, and can be formulated such that compounds having Formula I are stable, e.g., not reduced by antioxidant additives.

[00103] The additive may also comprise a thickening agent. Suitable thickening agents may be of those known and employed in the art, including, e.g., pharmaceutically acceptable polymeric materials and inorganic thickening agents. Exemplary thickening agents for use in the present pharmaceutical compositions include polyacrylate and polyacrylate co-polymer resins, for example poly-acrylic acid and poly-acrylic acid/methacrylic acid resins; celluloses and cellulose derivatives including: alkyl celluloses, e.g., methyl-, ethyl- and propyl-celluloses; hydroxyalkyl-celluloses, e.g., hydroxypropyl- celluloses and hydroxypropylalkyl-celluloses such as hydroxypropyl-methyl- celluloses; acylated celluloses, e.g., cellulose-acetates, cellulose- acetatephthallates, cellulose-acetatesuccinates and hydroxypropylmethyl- cellulose phthallates; and salts thereof such as sodium-carboxymethyl- celluloses; polyvinylpyrrolidones, including for example poly-N- vinylpyrrolidones and vinylpyrrolidone co-polymers such as vinylpyrrolidone- vinylacetate co-polymers; polyvinyl resins, e.g., including polyvinylacetates and alcohols, as well as other polymeric materials including gum traganth, gum arabicum, alginates, e.g., alginic acid, and salts thereof, e.g., sodium alginates; and inorganic thickening agents such as atapulgite, bentonite and silicates including hydrophilic silicon dioxide products, e.g., alkylated (for example methylated) silica gels, in particular colloidal silicon dioxide products.

[00104] Such thickening agents as described above may be included, e.g., to provide a sustained release effect. However, where oral administration is intended, the use of thickening agents may not be required. Use of thickening agents is, on the other hand, indicated, e.g., where topical application is foreseen.

[001051 In one embodiment of the invention, compounds having Formula I are formulated as described, for example, in U.S. patent 6,653,319.

[00106] Although the dosage of the compound having Formula I will vary according to the activity and/or toxicity of the particular compound, the condition being treated, and the physical form of the pharmaceutical composition being employed for administration, it may be stated by way of guidance that a dosage selected in the range from 0.1 to 20 mg/kg of body weight per day will often be suitable, although higher dosages, such as 0.1 to 50 mg/kg of body weight per day may be useful. Those of ordinary skill in the art are familiar with methods for determining the appropriate dosage. Methods for assessing the toxicity, activity and/or selectivity of the compounds having Formula I may be carried out using any of the methods known in the art, including the antiproliferative activity test.

[00107] In certain instances, the dosage of the compounds having Formula I will be lower, e.g., when used in combination with at least a second hyperproliferative disorder treatment, and may vary according to the activity and/or toxicity of the particular compound, the condition being treated, and the physical form of the pharmaceutical composition being employed for administration.

[00108] When the composition of the present invention is formulated in unit dosage form, the compound having Formula I will preferably be present in an amount of between 0.01 and 2000 mg per unit dose. More preferably, the amount of compound having Formula I per unit dose will be about 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, 1950, or 2000 mg or any amount therein.

(00109] When the unit dosage form of the composition is a capsule, the total quantity of ingredients present in the capsule is preferably about 10-1000 μL. More preferably, the total quantity of ingredients present in the capsule is about 100-300 μL. In another embodiment, the total quantity of ingredients present in the capsule is preferably about 10-1500 mg, preferably about 100- 1000 mg.

[00110] The relative proportion of ingredients in the compositions of the invention will, of course, vary considerably depending on the particular type of composition concerned. The relative proportions will also vary depending on the particular function of ingredients in the composition. The relative proportions will also vary depending on the particular ingredients employed and the desired physical characteristics of the product composition, e.g., in the case of a composition for topical use, whether this is to be a free flowing liquid or a paste. Determination of workable proportions in any particular instance will generally be within the capability of a person of ordinary skill in the art. AU indicated proportions and relative, weight ranges described below are accordingly to be understood as being indicative individually inventive teachings only and not as limiting the invention in its broadest aspect.

[00111] The amount of compound having Formula I in compositions of the invention will of course vary, e.g., depending on the intended route of administration and to what extent other components are present. In general, however, the compound having Formula I will suitably be present in an amount of from about 0.005% to 20% by weight based upon the total weight of the composition. In certain embodiments, the compound having Formula I is present in an amount of from about 0.01% to 15% by weight based upon the total weight of the composition.

[00112] In addition to the foregoing, the present invention also provides a process for the production of a pharmaceutical composition as hereinbefore defined, which process comprises bringing the individual components thereof into intimate admixture and, when required, compounding the obtained composition in unit dosage form, for example filling said composition into tablets, gelatin, e.g., soft or hard gelatin, capsules, or non-gelatin capsules.

[00113] The starting materials of the N-oxides of the present invention are known and described, for example, in U.S. patent Nos. 6,933,302, 6,893,668, 6,855,720, 6,825,207, 6,825,206, 6,821,982, 6,815,546, 6,809,103, 6,797,715, 6,794,370. 6,762,301, 6,759,416, 6,743,918, 6,743,917, 6,723,849, 6,723,729, 6,716,982, 6,699,876, RE38,408, 6,660,861, 6,629,995, 6,624,170, 6,620,937, 6,617,456, 6,608,076, 6,593,334, 6,589,939, 6,559,309, 6,548,488, 6,545,010, 6,512,118, 6,509,345, 6,506,734, 6,503,889, 6,500,953, 6,492,335, 6,486,320, 6,476,225, 6,476,043, 6,462,196, 6,455,699, 6,444,820, 6,436,951, 6,410,731, 6,407,118, 6,403,604, 6,403,603, 6,403,569, 6,376,676, 6,376,617, 6,352,996, 6,350,756, 6,313,135, 6,310,210, 6,310,207, 6,306,868, 6,288,072, 6,284,891, 6,281,223, 6,265,413, 6,252,079, 6,242,457, 6,239,278, 6,218,540, 6,218,399, 6,214,836, 6,211,371, 6,207,832, 6,194,579, 6,191,119, 6,177,568, 6,177,439, ,169,080,6,166,029, 6,156,897, 6,156,754, 6,150,343, 6,143,891, 6,136,978, 6,130,227, 6,121,451, 6,121,277, 6,107,486, 6,100,273, 6,096,336, 6,093,721, 6,080,751, 6,063,923, 6,057,303, 6,046,209, 6,040,313, 6,034,243, 6,028,078, 5,998,426, 5,990,120, 5,981,542, 5,972,955, 5,968,943, 5,932.709, 5,932,588, 5,922,877, 5,916,897, 5,916,896, 5,910,491, 5,892,043, 5,889,017, 5,880,133, 5,859,023, 5,859,022, 5,856,333, 5,843,954, 5,840,899, 5,837,673, 5,801,167, 5,786,344 ,5,773,522, 5,744,605, 5,736,156, 5,731,316, 5,726,181, 5,700,939, 5,677,286, 5,674,874, 5,674,873, 5,670,500, 5,663,177, 5,652,244, 5,646,159, 5,633,260, 5,614,628, 5,614,529, 5,608,066, 5,604,233, 5,602,141, 5,597,829, 5,559,235, 5,552,156, 5,552,154, 5,541,329, 5,541,327, 5,527,913, 5,525,731, 5,496,952, 5,496,830, 5,491,237, 5,486,615, 5,478,943, 5,475,108, 5,468,859, 5,468,754, 5,447,936, 5,446,047, 5,428,166, 5,405,963, 5,401,747, 5,395,939, 5,391,745, 5,364,858, 5,352,789, 5,342,947, 5,321,140, 5,315,007, 5,264,579, 5,258,516, 5,254,690, 5,247,089, 5,244,903, 5,243,050, 5,212,317, 5,200,524, 5,191,082, 5,180,722, 5,162,532, 5,122,526, 5,106,742, 5,061,800, 5,053,512, 5,049,668, 5,004,758, 4,981,968, 4,943,579, 4,939,255, 4,914,205, 4,894,456, RE32,518, 4,604,463, 4,545,880, 4,513,138, 4,473,692, 4,399,282, 4,399,276 and 4,031,098. The tertiary and/or heteroaromatic amine groups in camptothecin and analogs can selectively be oxidized using known oxidizing agents. Certain oxidizing agents that are known in the art for preparing the iV-oxides from aromatic and tertiary amine groups include, without limitation, potassium monopersulfate, monoperoxyphthalic acid, magnesium monoperoxyphthalate (MMPP), hydrogen peroxide, peracetic acid, trifluoroperacetic acid, perbenzoic acid, 3-chloroperbenzoic acid (CPBA), and 2-phenylsulfonyl-3- phenyloxaziridine (Davis reagent). The oxidation reaction can be carried out in a solvent such as chloroform, methylene chloride, 1 ,2-dichloroethane, or acetic acid, optionally in the presence of an alkali or alkaline-earth metal carbonate or bicarbonate. The reaction can be run from about 1 to 48 hours at a temperature of 0 ⁰C to reflux temperature, and checked periodically for the presence of the desired TV-oxide. Depending on the groups bound to the amine, reaction times may need to be adjusted accordingly to obtain appropriate quantities of the desired bis N-oxide product. See also Lee et al., "Nitracrine N-oxides: effects of variations in the nature of the side chain N- oxide on hypoxia-selective cytotoxicity" Anticancer Drug Des. 14(6):487-497 (1999).

[00115] In certain situations, more than one of the nitrogen atoms of the camptothecin analog may be oxidized simultaneously. In certain cases, one or more of the multiple N-oxide groups may be reduced selectively, leaving one or more of the other N-oxide groups in place. Thus, the present invention contemplates the preparation of N-oxide analogs in which one or more of the nitrogen atoms that are suitable for N-oxide formation are present as the N-oxide without regard to the susceptibility of a particular nitrogen atom to N-oxide formation or the susceptibility of a particular N-oxide group to reduction. It is envisaged to employ a combination of suitable protecting groups (see: Greene, T. W.; Wuts, P. G. M. Protective Groups in Organic Synthesis, second edition, Wiley Interscience, 1991) to protect those nitrogen atoms not undergoing oxidation.

[00116] By way of an example, primary and secondary amines that are present in a camptothecin analog may be protected using, for example, ter/-butyl sulfonyl (BUS) group. Jarowϊcki, K. and Kocienski, P., J. Chem. Soc, Perkin Trans 1, 4005-4037, 4029 (1998); Sun, P. and Weinreb, S. M. J. Org. Chem. (52:8604-08 (1997). The BUS protecting group is introduced by reaction of the amine with rerϊ-butylsulfinyl chloride followed by oxidation of the sulfinyl amide with, for example, dimethyldioxirane, m-chloroperbenzoic acid or RuCl₃ catalyzed NaIO₄. The oxidation step in the preparation of the BUS-protected primary or secondary amines may also oxide any tertiary amine and heteroaromatic nitrogen present in the compounds. Thus, this protecting group may be introduced into primary and secondary amines while simultaneously oxidizing tertiary and heteroaromatic nitrogen atoms. [00117] The BUS protecting group is stable towards strong reagents such as alkyllithium, Grignard reagents, 0.1M HCl in MeOH (20° C, 1 hr), 0.1M TFA in dichloromethane (20° C, 1 hr) and pyrolysis at 180° C. The BUS-protected secondary amines can be cleaved with 0.1 M triflic acid in dichloromethane containing anisole as a cation scavenger at 0° C for 15-30 minutes while primary amines are released more slowly at room temperature. If desired, both BUS-protected primary and secondary amines may be deprotected with 0.1 M triflic acid in dichloromethane containing anisole as a cation scavenger at 25° C for 2.5 hours, Thus, the BUS protecting group may allow protecting primary and secondary amines simultaneously while also oxidizing tertiary amines and heteroaromatic nitrogen atoms to the N-oxides. Moreover, the BUS protecting group may allow protecting primary and secondary amines simultaneously, oxidizing tertiary amines and heteroaromatic nitrogen atoms to N-oxides, deprotect the secondary amine selectively, alkylate the secondary amine to a tertiary amine, oxidize the resulting tertiary amine and deprotect the primary amine. Alternatively, a primary and a secondary amine may be protected with BUS protecting group, the secondary amine may be deprotected selectively, the secondary amine may be protected with, for example, Boc protecting group, and then the primary amine may be deprotected selectively followed by alkylation and oxidation. Thus, when a primary amine and a secondary amine are present in a camptothecin analog, a BUS protecting group may be used to transform one of amines to an N-oxide without affecting the other.

[00118J Recent development in the use of Boc group to protect amines allows introduction and removal of the group under mild conditions. For example, a camptothecin analog amine group may be protected with Boc group by simply mixing the analog and Boc-ON (2-(Boc-oxyimino)-2-phenylacetonitrile, available from Aldrich Co.) in benzene at 25° C for 20 minutes (or 6 hours if the amine is an electron deficient aniline) in the presence of powdered zinc. See Spivey, A. C. and Maddaford A. Anna. Rep. Prog. Chem., Sect. B, 95:83- 95 (1999). Alkyl esters are tolerated. [00119] Boc-protected amines are generally deprotected using triflic acid although recent developments generally use mildly acidic conditions that leave acid-labile groups unaffected. For example, heating Boc protected p-anisidines at 180° C in the presence of 4-chlorophenol deprotects the amine group without affecting acid sensitive methoxy enols (-CH=C(OCH₃)-). Jarowicki, K. and Kocienski, P., J. Chem. Soc, Perkin Trans 1, 4005-4037, 4025 (1998). Thus, primary and secondary amines in camptothecin may be protected with Boc group followed by oxidation of the tertiary amines and deprotection of the primary and secondary amines.

[001201 It has also recently been reported a new base-sensitive amino protecting group l,l-dioxobenzo[&]thiophene-methoxycarbonyl (Bsmoc). Bsmoc is introduced via its chloroformate or N-hydroxy-succinimide derivative. The Bsmoc group is stable towards tertiary amines for 24 hours but is removed within 3-5 minutes using piperidine. Jarowicki, K. and Kocienski, P., J. Chem. Soc, Perkin Trans 1, 4005-4037, 4027 (1998). Thus, primary and secondary amines present in camptothecin analogs may conveniently be protected with Bsmoc protecting group followed by oxidation of the tertiary amines and removal of the protecting group under mild conditions.

[00121] It has also been reported that certain heteroaromatic nitrogen atoms can be oxidized selectively in the presence of certain aromatic primary amines and certain secondary amines adjacent to a double bond. Delia, T. J. et al. J. Org. Chem. 30:2766-68 (1965). For example, oxidation of cytosine with m-chloroperbenzoic acid results in cytosine 3-N-oxide despite the presence of aromatic primary amine and a secondary amine. Thus, heteroaromatic nitrogen atoms and tertiary amines may be oxidized in the presence of certain aromatic primary amines and secondary amines.

[00122] The following examples are illustrative, but not limiting, of the method and compositions of the present invention. Other suitable modifications and adaptations of the variety of conditions and parameters normally encountered in clinical therapy and which are obvious to those skilled in the art are within the spirit and scope of the invention.

EXAMPLES

[00123] ¹H NMR spectra were obtained from INOVA-300 MHz or INOVA-

500 MHz spectrometers using CDCI3 as a solvent (s, d, t, dd and m indicate singlet, doublet, triplet, doublet of doublet, and raultiplet, respectively). Analytical thin-layer chromatography was performed on polyester-backed plates from EMD Chemicals, Inc., precoated with silica gel 60 F2₅₄. Radial thin-layer chromatography was performed on a Harrison Research Chromatron (7924T) using 2 mm thick silica plates coated in our laboratory with silica gel 60 PF254 containing gypsum (EMD Chemicals, Inc.), Analytical scale high- performance liquid chromatography (HPLC) was performed on a 4.6 mm x 250 mm MICROSORB Ci₈ column using a pressure of 1800-2100 psi.

EXAMPLE 1

OXIDATION OF 20(S) -CAMPTOTHECIN

[00124] 3.0 mL of dimethyldioxirane (3.0 mL) was added in excess to 20(S)- camptothecin (20.0 mg, Aldrich, used as received, Lot #1621 ILI) in a tared, 10 mL, round-bottomed flask. The reaction was stirred for an hour at 0⁰C. The progress of the reaction was monitored by TLC (10:0.35 CH₂Cl₂: CH₃ OH). The solution was concentrated at reduced pressure. Crystallization of 20(S)-Camptothecin iV-oxide occurred from 4:1 ethyl acetate:hexanes. Recrystallization from 4:1 chloroform:hexanes yielded a yellow-orange powder. Physical properties are as follows: Rf (20(S)-Camptothecin, 0.70; 20(S)-Camptothecin N-oxide, 0.30), mp 254-256°C, [Lit. 254⁰C]. EXAMPLE 2

OXIDATION OF 7-ETHYL-10-HYDROXY CAMPTOTHECIN (SN-38)

[00125] 7-Ethyl-lO-hydroxycamptothecin (200 mg, 0.51 mmol, used as received from Abatra, COA No.0407051) was dissolved in 44 mL of glacial acetic acid. To this solution, 3.4 rnL of 30% H₂O₂ was added. The solution was heated at 70-80⁰C for 3 hours. TLC (10:1 CH₂Cl₂ICH₃OH)^' was used to monitor the reaction to completion. The solution was removed from heat and concentrated at low pressure at 45-50⁰C. The resulting dark orange liquid was dissolved in methanol (about 30 mL) and the desired product precipitated out of the solution as an orange powder. The supernatant liquid was decanted and the residue was washed with ether (2 x 5 mL), and then dried under vacuum to give SN-38 N-oxide (70 mg, 33%) as an orange powder, which was pure by TLC and NMR. The purity was 96% (area under the curve) by HPLC analysis. ¹H-NMR (DMSO-d₆) δ: 0.86 (t, J - 7.5, 3H), 1.26 (t, J = 7.5, 3H)₅ 1.78-1.95 (m, 2H)₃ 3.00 (q, J = 7.5, 2H), 5.28 (s, 2H), 5.41 (s, 2H)₅ 6.50 (s, IH)₅ 7.40 (s, IH), 7.41 (dd, J = 2, 9.3, IH), 8.02 (s, IH), 8.55 (d, J = 9.3, IH), 10.62 (s, IH).

EXAMPLE 3

CYTOTOXICITY OF CAMPTOTHECIN AND ANALOG N-OXIDES THEREOF IN LYMPHOMA₅ LEUKEMIA, AND MULTIPLE MYELOMA

[00126] The cytotoxicity of camptothecin and analog N-oxides thereof on different lymphoma, leukemia, and multiple myeloma cell lines will be tested in vitro under normoxic as well as 0.1-5% O2 hypoxic conditions. Standard cytotoxicity assays using MTS dye will be run to determine the IC50 f°^{r eacn} compound. Cells will be exposed to the compounds for 24, 48 or 72 hours and cells will be stained 24-72 hours post-drug exposure. Positive controls will use chemotherapeutic agents at doses shown in the art to be effective. The results should indicate that camptothecin analogs are cytotoxic to many of the cell lines, with IC50 values in the nanomolar to sub-nanomolar range.

Camptothecin analog N-oxides are expected to be less active or inactive compared to non-N-oxide under normoxic conditions. However, under 0.1-5% O2 hypoxic conditions, camptothecin analog N-oxides are expected to be converted to the corresponding parent non-N-oxide, which are expected to be cytotoxic with IC50 values in the millimolar to sub-nanomolar range.

EXAMPLE 4

CYTOTOXICITY OF CAMPTOTHECIN ANALOGS AND N-OXIDES THEREOF IN SOLID TUMOR LINES

[00127] The cytotoxicity of camptothecin and analog N-oxides thereof on different solid tumor cell lines will be tested ^" in vitro under normoxic conditions and 0.1-5% O2 hypoxic conditions. Standard cytotoxicity assays using MTS dye will be run to determine the IC50 for each compound. Cells will be exposed to the compounds for 24, 48 or 72 hours and cells will be stained 24-72 hours post-drug exposure. Chemotherapeutic agents at doses shown in the art to be effective will be used as positive controls. The results are expected to indicate that camptothecin analogs are cytotoxic to many of the cell lines, with IC50 values in the nanomolar to sub-nanomolar range.

Camptothecin and analog N-oxides thereof are expected to be less active or inactive compared to the corresponding camptothecin analogs.

EXAMPLE 5

ANTIPROLIFERATIVE ACTIVITY OF CAMPTOTHECIN AND ANALOG N-OXIDES THEREOF IN CANCER CELLS

[00128] The anti-proliferative activity of camptothecin and analog N-oxides thereof on established and primary tumor cell lines will be tested in vitro under normoxic and 0.1-5% O2 hypoxic conditions at concentrations ranging from 1 nM to 10 mM. The an ti -proliferative effect will be measured using the 5-bromo-2'-deoxyuridine ("BrDU") incorporation technique. The cells will be exposed to the compounds in the presence of BrDU for 24, 48 or 72 hours. BrDU is incorporated into the replicating cellular DNA. After cell fixation and washing, the incorporated BrDU is determined in a specific ELISA using an antibody specific to BrDU coupled to peroxidase. The N-oxides are expected not to have significant anti-proliferative activity in cancer cells at concentrations of up to 10 mM under normoxia. However, the N-oxides are expected to exhibit significant anti-proliferative effect on the cancer cell lines under 0.1-5% O2 hypoxia.

EXAMPLE 6

Anti-Tumor Activity of Camptothecin and Analog N-Oxide in Murine

Tumor Models

[00129] The in vivo antitumor efficacy of camptothecin and analog N-oxides will be evaluated using xenograft murine models. For example, male 5 to 6 week old nude mice will be inoculated subcutaneously in the mammary fat pad on each side with an injection of a human cancer cell line, for example about 1x10⁶ MDA-MB-231 (2LMP) in 0.3 ml serum free medium. The best xenograft recipients will be used. Treatments with camptothecin and analog N-oxides will begin when tumors averaged about 5-7 mm in diameter and will be continued for 4 weeks with a 2 month follow up period.

[00130] Test animals will be divided into cohort groups of 5-8 animals into the following treatment groups: Control (Group 1); Vehicle Control (Group 2), daily administration of the vehicle only; camptothecin and analog N-oxide (Group 3), 0.1 mg/kg administered orally per day for 4 weeks; Camptothecin and analog N-oxide (Group 4), 1 mg/kg administered orally per day for 4 weeks; Camptothecin and analog N-oxide (Group 5), 10 mg/kg administered orally per day for 4.

[00131] The test doses may further be increased or decreased upon recognition that such a modification is warranted for a particular camptothecin analog N-oxide. For example, a particular camptothecin analog N-oxide may exert its own, for example, antiproliferative effect, making it more effective than it would have been had camptothecin analog N-oxide been biologically inactive. In such a circumstance, lower doses of the camptothecin analog N-oxide may prove effective. Moreover, more cohort groups may be added to test additional doses of various camptothecin analog N-oxides.

[00132] During the treatment course, tumor sizes and animal weights will be measured periodically, for example 1-3 times per week for each animal. Following treatment, tumor sizes will be measured periodically, for example, 1-3 times per week, and animal weights once per week. Tumor sizes and weight observations will be made without knowledge of the animal's treatment group.

[00133] The data will be modeled using, for example, the natural logarithm of tumor volume versus time. Comparisons of the growth rates of animals administered with varying dosages will be performed. For example, Tumor Growth Delay and Tumor Growth Inhibition for varying dosages will be compared to the control.

[00134] Further, the in vivo antitumor activity of camptothecin and analog

N-oxide in combination with chemotherapeutic agents and/or radiotherapy will be evaluated using a xenograft model in nude mice.

[00135] Having now fully described this invention, it will be understood by those of ordinary skill in the art that the same can be performed within a wide and equivalent range of conditions, formulations and other parameters without affecting the scope of the invention or any embodiment thereof. All patents, patent applications and publications cited herein are fully incorporated by reference herein in their entirety.

Claims

WHAT IS CLAIMED IS:

1. A compound of Formula I:

or a pharmaceutically acceptable salt or prodrug thereof, wherein:

R' is hydrogen,Ri-O-(CH₂)_m-CO or Ri₆PO₂H-, m is an integer of 0-10; and

Ri is lower alkyl; phenyl optionally substituted with from one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, formyl, lower alkyl carbonyl, hydroxycarbonyl, lower alkylcarbonyloxy, benzyloxy, optionally substituted piperazino, lower alkoxycarbonyl, and lower alkylcarbonylamino; cycloalkyl of 3-7 carbons, optionally substituted with one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino; a fused, 2-, 3-, or 4-ring heterocyclic system optionally substituted with one to five substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, ammo, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino;

1- or 2-naphthyl optionally substituted with from one to four substituents independently selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino; a 5 or 6 membered heterocyclic ring containing one or two nitrogen atoms, which ring is optionally substituted with one or two substituents selected from the group consisting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, and lower alkylcarbonylamino;

R₂ is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, R¹C(O)O, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, -C(O)H, lower alkoxycarbonyl, tri-lower alkylsilyl, lower alkylcarbonyloxy, lower alkylcarbonylamino, lower alkylcarbonyloxymethyl, substituted vinyl, l-hydroxy-2-nitroethyl, alkoxycarbonylethyl, aminocarbonyl, mono- or di-alkylcarbonyl, alkylcarbonyloxymethyl, benzoylmethyl, benzylcarbonyloxymethyl, or mono- or di lower alkoyxymethyl.

R₃ is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, R¹C(O)O, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, CH₂NR₇RsRi₂ (where each of R₇ and R₈ is independently H-, alkyl of 1-6 carbons, optionally substituted phenyl, hydroxy lower alkyl, amino lower alkyl, or mono- or dialkylamino lower alkyl, or R₇ and Rg taken together with -N- represent a cyclic amino-), -C(O)H, -CH₂R₉ (where R₉ is lower alkoxy, CN, amino lower alkoxy., mono- or di- lower alkylamino lower alkoxy, lower alkylthio, amino lower alkylthio, or mono- or di-lower alkylamino lower alkylthio), or NRioRπRi₂ (where each of Rio and Rn is independently hydrogen, lower alkyl, phenyl, hydroxy lower alkyl or amino lower alkyl, or R₁₀ and Ru taken together with -N- represent a cyclic amino), dialkylamino alkyl, lower alkylcarbonyloxy, or lower alkylcarbonylamino;

R₄ is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy cyano, nitro, amino, amino lower alkyl, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, carbamoyloxy, lower alkylcarbonyloxy, lower alkylcarbonylamino, or Ri₇COO- wherein Ri₇ is a 5 or 6 membered heterocyclic ring containing one or two nitrogen atoms and which is substituted with obe or two substituents selected from the group consisiting of halo, lower alkyl, lower alkoxy, hydroxy, cyano, nitro, amino, NR₁0R1₁R12 (where each of Rio and Rn is independently H-, lower alkyl, phenyl, hydroxy lower alkyl, amino lower alkyl, or Rio and R_H taken together with -N- represent a cyclic amino-), dialkylamino alkyl, lower alkylcarbonyloxy, or lower alkylcarbonylamino; or R₄ together with R₃ is methylenedioxy;

R₅ is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, R¹C(O)O, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, or lower alkylcarbonylamino; and

Rn is hydrogen, halo, lower alkyl, lower alkoxy, hydroxy, R¹C(O)O, cyano, nitro, amino, halogenated lower alkyl, halogenated lower alkoxy, hydroxcarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, or lower alkylcarbonylamino;

R₁₆ is hydroxy, alkyl, alkoxy or phenyl; and each Of R]₂ and Ri₅ is O or is absent provided that at least one Of Ri₂

2. The compound of claim 1 , wherein each of R₃, R₅ and R₆ is hydrogen.

3. The compound of claim 1, wherein said compound of Formula I is selected from the group consisting of N-oxides and bis N-oxides of: (7-ethyl-10-[4-(l-piperidino)-l-piperidino]carbonyloxy)-camptothecin; 9-nitro camptothecin; 7-chloro camptothecin; 7-n-propyldimethyisilyl camptothecin; 10-hydroxy-7-n-ρropyldimethylsilyl camptothecin; 10-hydroxy-7-chlorocamptothecin; ' 10-acetoxy-7-n-propyldimethylsilyl camptothecin; lO-acetoxy-7-chlorocamptothecin; 7-tert-butyldimethylsilyl camptothecin; 10-hydroxy-7-tert-butyldimethylsilyl camptothecin; 10-acetoxy-7-tert-butyldimethylsilyl camptothecin; 9-hydroxy camptothecin; 9-amino camptothecin; 10-amino camptothecin; 9-amino-l 0-hydroxy camptothecin; 9-amino-10,l 1-methylenedioxy camptothecin; 9-methylamino camptothecin; 9-methyl camptothecin; 9-dimethy laminomethyl camptothecin ; 9-chloro camptothecin; 9-fluoro camptothecin; 9-piperidino camptothecin;

9-dimethylaminomethyl-l 0-hydroxy camptothecin; 9-morpholinomethyl camptothecin; 10-hydroxy camptothecin; 9,10-dichloro camptothecin; 10-bromo camptothecin; 10-chloro camptothecin; 0-methyl camptothecin; 0-fluoro camptothecin; 0-nitro camptothecin; 0,11 -methyl enedioxy camptothecin; 0-formyl camptothecin; 0-nonylcarbonyloxy camptothecin;

10-undecylcarbonyloxy camptothecin;

10-pentadecylcarbonyloxy camptothecin;

10-heptadecylcarbonyloxy camptothecin;

10-nonadecylcarbonyloxy camptothecin; -nitro- 10, 11 -methylenedioxy camptothecin; -(4-methylpiperazinylmethyl)- 10-hydroxy camptothecin; -[4-(l -piperidino)-l-piperidinomethyl]-10-hydroxy camptothecin; -methyl-10,l 1 -methylenedioxy camptothecin; -chloro-10,l 1 -methylenedioxy camptothecin; -cyano-10,l 1 -methylenedioxy camptothecin;

9-acetoxy-l 0,11 -methylenedioxy camptothecin;

9-acetylamino-10,l 1 -methylenedioxy camptothecin;

9-aminomethyl-l 0-hydroxy camptothecin;

9-ethoxymethyl-l 0-hydroxy camptothecin;

9-methylaminomethyl-l O-hydro^fy camptothecin;

9-n-propylaminomethyl-10-hydroxy camptothecin;

9-dimethylaminomethyl- 10-hydroxy camptothecin;

9-cyclohexylaminomethyl- 10-hydroxy camptothecin;

9-(2-hydroxyethyl)aminomethyl- 10-hydroxy camptothecin;

9-(trimethylammonio)methyl- 10-hydroxy camptothecin, methanesulfonate;

9-moφholinom ethyl- 10-hydroxy camptothecin;

9-cyanomethyl-lO-hydroxy camptothecin; camptothecin-7-aldehyde;

10-methoxy camptothecin-7-aldehyde;

7-acetoxymethyl camptothecin; -acetoxymethyl-l 0-methyl caπiptothecin; -cyano-lO-raethoxy camptothecin; -cyano camptothecin; -formyJethenyl camptothecin; -ethoxycarbonylethenyl camptothecin; -cyanoethenyl camptothecin;

7-(2,2-dicyanoethenyl) camptothecin;

7-(2-cyano-2-ethoxycarbonyl)ethenyl camptothecin;

7-ethoxycarbonyl ethyl camptothecin ;

7-ethyl camptothecin;

7-n-propyl camptothecin;

7-acetoxymethyl camptothecin;

7-n-propylcarbonyloxymethyl camptothecin;

7-ethoxycarbonyl camptothecin;

7-ethyl- 10-hydroxy camptothecin;

7-ethyl- 10-acetyloxy camptothecin;

7-methyl-l O-aminocarbonyloxy camptothecin;

7-n-propyl-l O-piperidinocazbonyloxy camptothecin;

7-ethyl- 10-(2-dimethylamino)ethyl camptothecin;

7-ethyl- 10-[4(l-piperidino)-piperidino carbonyloxy camptothecin;

7-ethyl- 10-(l -piperazine)carbonyloxy camptothecin;

7-ethyl- 10-(4-i-proρylaminocarbonylmethylpiperazine)carbonyloxy camptothecin;

7-ethyl- 10-[4(l -pyrrolidinyl)piperazine]carbonyloxy camptothecin;

7-ethyl-10-[(4-(dimethylamino)-l-ρiperidino]carbonyloxy camptothecin;

7-ethyl- 10-[4-(di-n-propylamino)- 1 -piperidinol]carbonyloxy camptothecin;

7-ethyl- 10-[(4-(di-n-butylamino)- 1 -piperidino]carbonyloxy camptothecin;

7-ethyl-10-[4-(l-pyrrolidino)-l-piperidino)]carbonyloxy camptothecin;

7-ethyl-10-[4-(l-piperidino)-l-piperidino]carbonyloxy camptothecin; and

7-ethyl-10-[N-methyl-N-2-(dimethylamino)ethylamino]carbonyloxy camptothecin; wherein said compound has 20(S) or 20(RS) configuration.

4. The compound of claim 1 , wherein said compound of Formula I is selected from the group consisting of 9-dirnethylaminornethyl- 10-hydroxycamptothecin N-oxide, 9-dimethylaminornethyl- 10-hydroxycamptothecin bis N-oxide, 7-ethyl-10[4-(l-piperidino)- l-piperidino]carbonyloxycamptothecin N-oxide, 7-ethyl-10[4-(l-piperidino)- l-piperidino]carbonyloxycamptothecin bis N-oxide, 9-aminocamptothecin N-oxide, 10-aminocamptothecin N-oxide, 10,11-methylenedioxycamptothecin N-oxide and 7-ethyl-lO-hyrdoxycamptothecin N-oxide or a pharmaceutically acceptable salt or prodrug thereof.

5. A method of treating, ameliorating, or preventing a hyperproliferative disorder comprising administering to an animal in need thereof a therapeutically effective amount of a compound of claim 1.

6. A method of treating, preventing or ameliorating a hyperproliferative disorder in an animal in need thereof, comprising

(a) determining whether said hyperproliferative disorder is characterized by hypoxic tissue, and

(b) treating said animal with an effective amount of a compound of claim 1.

7. The method of claim 5 or 6, wherein said compound of Formula I is selected from the group consisting of N-oxides and bis N-oxides of:

(7-ethyl-l 0-[4-(l -piperidino)-l -piperidino]carbonyloxy)-camptothecin;

9-nitro camptothecin;

7-chloro camptothecin;

7-n-propyldimethylsilyl camptothecin;

10-hydroxy-7-n-ρroρyldimethylsilyl camptothecin; 10-hydroxy-7-chlorocamptothecin; lO-acetoxy-7-n-propyldiraethylsilyl camptothecin;

10-acetoxy-7-chlorocaraptothecin;

7-tert-butyldim ethyl si IyI camptothecin ;

10-hydroxy-7-tert-butyldimethylsilyl camptothecin;

10-acetoxy-7-tert-butyldimethylsilyl camptothecin;

9-hydroxy camptothecin;

9-amino camptothecin;

10-amino camptothecin;

9-amino-10-hydroxy camptothecin;

9-amino-10,l 1 -methyl enedioxy camptothecin;

9-methylamino camptothecin;

9-methyl camptothecin;

9-dimethylaminomethyl camptothecin;

9-chloro camptothecin;

9-fluoro camptothecin;

9-piperidino camptothecin;

9-dimethylaminomethyl- 10-hydroxy camptothecin;

9-morpholinomethyl camptothecin;

10-hydroxy camptothecin;

9,10-dichloro camptothecin;

10-bromo camptothecin;

10-chloro camptothecin;

10-methyl camptothecin;

10-fluoro camptothecin;

10-nitro camptothecin;

10, 11 -methylenedioxy camptothecin;

10-formyl camptothecin;

10-nonylcarbonyloxy camptothecin;

10-undecylcarbonyloxy camptothecin;

10-pentadecylcarbonyloxy camptothecin; 0-heptadecylcarbonyloxy camptothecin; 0-nonadecylcarbonyloxy camptothecin; -nitro-10,l 1-methylenedioxy camptothecin; -(4-methylpiperazinylmethyl)- 10-hydroxy camptothecin; -[4-(l -piperi dino)- 1 -piperidinomethyl]- 10-hydroxy camptothecin; -m ethyl- 10,11-methylenedioxy camptothecin; -chloro- 10,11-methylenedioxy camptothecin; -cyano-10,l 1-methylenedioxy camptothecin; -acetoxy-10,l 1-methylenedioxy camptothecin; -acetylamino- 10,11 -methylenedioxy camptothecin; -aminomethyl- 10-hydroxy camptothecin;

9-ethoxymethyl-l 0-hydroxy camptothecin;

9-methylarainomethyl-l 0-hydroxy camptothecin;

9-n-propylaminomethyl-l 0-hydroxy camptothecin;

9-dimethylaminomethyl- 10-hydroxy camptothecin;

9-cyclohexylaminomethyl- 10-hydroxy camptothecin;

9-(2-hydroxyethyl)aminomethyl-10-hydroxy camptothecin;

9-(trimethylammonio)methyl-l 0-hydroxy camptothecin, methanesυlfonate;

9-morpholinomethyl- 10-hydroxy camptothecin;

9-cyanomethyl- 10-hydroxy camptothecin; camptothecin-7-aldehyde;

10-methoxy camptothecin-7-aldehyde;

7-acetoxymethyl camptothecin;

7-acetoxym ethyl- 10-methyl camptothecin;

7 -cyano- 10-methoxy camptothecin ;

7-cyano camptothecin;

7-formylethenyl camptothecin;

7 -ethoxycarbonylethenyl camptothecin ;

7-cyanoethenyl camptothecin;

7-(2,2-dicyanoethenyl) camptothecin;

7-(2-cyano-2-ethoxycarbonyl)ethenyl camptothecin; -ethoxycarbonylethyl camptothecin;

7-ethyl camptothecin; ^'

7-n-propyl camptothecin;

7-acetoxymethyl camptothecin;

7-n-propylcarbonyloxymethyl camptothecin;

7-ethoxycarbonyl camptothecin;

7-ethyl- 10-hydroxy camptothecin;

7-ethyl- 10- acetyloxy camptothecin;

7-methyl-l 0-aminocarbonyloxy camptothecin;

7-n-propyl- 10-piperidinocazbohyloxy camptothecin;

7-ethyl-l 0-(2-dimethylamino)ethyl camptothecin;

7-ethyl-10-[4(l-piperidino)-piperidino carbonyloxy camptothecin;

7-ethyl- 10-(l -piperazine)carbonyloxy camptothecin;

7-ethyl- 10-(4-i-propylaminocarbonylmethylpiperazine)carbonyloxy camptothecin;

7-ethyl-10-[4(l-ρyrrolidinyl)piperazine]carbonyloxy camptothecin;

7-ethyl- 10-[(4-(dimethylamino)-l -piperidino]carbonyloxy camptothecin;

7-ethyl-10-[4-(di-n-propylamino)-l-piperidinol]carbonyloxy camptothecin;

7-ethyl- 10-[(4-(di-n-butylamino)- 1 -piperidino]carbonyloxy camptothecin;

7-ethyl-10-[4-(l-pyrrolidino)-l-piperidino)]carbonyloxy camptothecin;

7-ethyl-l 0-[4-(l -piperidino)-l -piperidino] carbonyloxy camptothecin; and

7-ethyl-10-[N-methyl-N-2-(dimethylamino)ethylarnino]carbonyloxy camptothecin; or a pharmaceutically acceptable salt or prodrug thereof, wherein said compound has 20(S) or 20(RS) configuration.

8. The method of claim 5, further comprising subjecting said animal to an imaging technique selected from the group consisting of computed tomography, magnetic resonance imaging, single photon emission computer tomography and positron emission tomography prior to or during administration of said compound.

9. The method of claim 5 or 6, wherein said hyperproliferative disorder is cancer.

10. The method of claim 9, wherein the cancer is of the bladder, brain, breast, cervix, colon, endometrium, esophagus, head and neck, kidney, larynx, liver, lung, oral cavity, ovaries, pancreas, prostate, skin, stomach, or testis.

11. The method of claim 9, wherein the cancer is selected from the group consisting of acute and chronic lymphocytic leukemia, acute granulocytic leukemia, adrenal cortex carcinoma, bladder carcinoma, breast carcinoma, cervical carcinoma, cervical hyperplasia, choriocarcinoma, chronic granulocytic leukemia, chronic lymphocytic leukemia, colon carcinoma, endometrial carcinoma, esophageal carcinoma, essential thrombocytosis, genitourinary carcinoma, hairy cell leukemia, head and neck carcinoma, Hodgkin's disease, Kaposi's sarcoma, lung carcinoma, lymphoma, malignant carcinoid carcinoma, malignant hypercalcemia, malignant melanoma, malignant pancreatic insulinoma, medullary thyroid carcinoma, melanoma, multiple myeloma, mycosis fungoides, myeloid and lymphocytic leukemia, neuroblastoma, non-Hodgkin's lymphoma, ■ osteogenic sarcoma, ovarian carcinoma, pancreatic carcinoma, polycythemia vera, primary brain carcinoma, primary macroglobulinemia, prostatic carcinoma, renal cell carcinoma, rhabdomyosarcoma, skin cancer, small-cell lung carcinoma, soft-tissue sarcoma, squamous cell carcinoma, stomach carcinoma, testicular carcinoma, thyroid carcinoma, and Wilms' tumor.

12. The method of claim 5 or 6, wherein said hyperproliferative disorder is age-related macular degeneration, Crohn's disease, cirrhosis, chronic inflammatory-related disorders, proliferative diabetic retinopathy, proliferative vitreoretinopathy, retinopathy of prematurity, granulomatosis, immune hyperproliferation associated with organ or tissue transplantation, an immunoproliferative disease or disorder, psoriasis, rheumatoid arthritis, systemic lupus erythematosus, vascular hyperproliferation secondary to retinal hypoxia, or vasculitis.

13. The method of claims 5 or 6, further comprising administering one or more' other active agents or treatments to the animal.

14. The method of claim 13, wherein said one or more other active agents or treatments are independently selected from the group consisting of a chemotherapeutic agent, a radiotherapeutic agent/treatment, an anti-angiogenesis agent, a vascular targeting agent, an HIFl inhibitor, an Hsp90 inhibitor, a tyrosine kinase inhibitor, a serine/threonine kinase inhibitor, a proteasome inhibitor, an HDAC inibitor, a caspase inducer, a CDK inhibitor, and a proapoptotic molecule.

15. The method of claim 14, wherein the chemotherapeutic agent is selected from, the group consisting of abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic trioxide, asparaginase, BCG live, bevaceizumab, bexarotene, bleomycin, bortezomib, busulfan, calusterone, camptothecin, capecitabine, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cinacalcet, cisplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, darbepoetin alfa, daunorubicin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone, Elliott's B solution, epirubicin, epoetin alfa, estramustine, etoposide, exemestane, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gemcitabine, gemtuzumab ozogamicin, gefitinib, goserelin, hydroxyurea, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib, interferon alfa-2a, interferon alfa-2b, irinotecan, letrozole, leucovorin, levamisole, lomustine, meclorethamine, megestrol, melphalan, mercaptopurine, mesna, methotrexate, rnethoxsalen, methylprednisolone, mitomycin C, mitotane, mitoxantrone, nandrolone, nofetumomab, oblimersen, oprelvekin, oxaliplatin, paclitaxel, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed, pentostatin, pipobroman, plicamycin, polifeprosan, porfimer, procarbazine, quinacrine, rasburicase, rituximab, sargramostim, streptozocin, talc, tamoxifen, tarceva, temozolomide, teniposide, testolactone, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, and zoledronate.

16. The method of claim 14, wherein said anti-angiogenesis agent is selected from the group consisting of bevacizumab, angiostatin, endostatin, batimastat, captopril, cartilage derived inhibitor, genistein, interleukin 12, lavendustin, medroxypregesterone acetate, recombinant human platelet factor 4, tecogalan, thrombospondin, TNP-470, anti-VEGF monoclonal antibody, soluble VEGF-receptor chimaeric protein, anti-VEGF receptor antibodies, anti-PDGF receptors, inhibitors of integrins, tyrosine kinase inhibitors, serine/threonine kinase inhibitors, antisense oligonucleotides, antisense oligodexoynucleotides, siRNAs, anti-VEGF aptamers and pigment epithelium derived factor.

17. The method of claim 13, wherein said compound is administered prior to the administration of said active agents or treatments.

IS. The method of claim 13, wherein said compound is administered concurrently with the administration of said active agents or treatments.

19. The method of claim 18, wherein the administration of said compound is continued beyond the administration of said active agents or treatments.

20. ^■ The method of claim 13, wherein said compound is administered after the administration of said active agents or treatments.

21. The method of claim 13, wherein the method is repeated at least once.

22. A pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically acceptable carrier.

23. The pharmaceutical composition of claim 22, wherein said compound is selected from the group consisting of N-oxides and bis N-oxides of:

(7-ethyl-10-[4-(l-piperidino)-l-piperidino]carbonyloxy)-camptothecin; 9-nitro camptothecin;

7-chloro camptothecin;

7-n-propyldimethylsilyl camptothecin; lO-hydroxy-7-n-propyldimethylsilyl camptothecin;

10-hydroxy-7-chloro camptothecin;

10-acetoxy-7-n-propyldimethylsilyl camptothecin;

10-acetoxy-7-chlorocamptothecin;

7-tert-butyldimethylsilyl camptothecin;

10-hydroxy-7-tert-butyldimethylsilyl camptothecin; lO-acetoxy-7-tert-butyIdimethylsilyl camptothecin;

9-hydroxy camptothecin;

9-amino camptothecin;

10-amino camptothecin;

9-amino- 10-hydroxy camptothecin;

9-amino- 10,11-methylenedioxy camptothecin;

9-methylamino camptothecin;

9-m ethyl camptothecin;

9-dimethylaminomethyl camptothecin; -chloro camptothecin; -fluoro camptothecin; -piperidino camptothecin; -dimethylaminomethyl-l O-hydroxy camptothecin; -morpholinomethyl camptothecin; O-hydroxy camptothecin; ,10-dichloro camptothecin; 0-bromo camptothecin; 0-chloro camptothecin; 0-methyI camptothecin; 0-fluoro camptothecin; 0-nitro camptothecin; 0, 11 -methylenedioxy camptothecin; 0-formyl camptothecin; 0-nonylcarbonyloxy camptothecin; 0-undecylcarbonyloxy camptothecin; 0-pentadecylcarbonyloxy camptothecin; 0-heptadecylcarbonyloxy camptothecin; 0-nonadecylcarbonyloxy camptothecin; -nitro- 10,11 -methylenedioxy camptothecin; -(4-methylpiperazinylmethyl)- 10-hydroxy camptothecin; -[4-(l-piperidino)-l -piperidinomethyl]-l O-hydroxy camptothecin; -methyl-10,l 1 -methylenedioxy camptothecin; -chloro-10,l 1 -methylenedioxy camptothecin; -cyano-10,l 1 -methylenedioxy camptothecin; -acetoxy-10,l 1 -methylenedioxy camptothecin; -acetylamino-10,l 1 -methylenedioxy camptothecin; -aminomethyl-l O-hydroxy camptothecin; -ethoxymethyl-l 0-hydroxy camptothecin; -methylaminomethyl-l O-hydroxy camptothecin; -n-propylaminomethyl-l O-hydroxy camptothecin; 9-dirnethylaminomethyl-l 0-hydroxy camptothecin;

9-cyclohexylaminomethyl- 10-hydroxy camptothecin;

9-(2-hydroxyethyl)aminomethyl- 10-hydroxy camptothecin ;

9-(trimethylaτnmonio)methyl- 10-hydroxy camptothecin, methanesulfonate;

9-morpholinomethyl- 10-hydroxy camptothecin;

9-cyanomethyl- 10-hydroxy camptothecin; camptothecin-7-aldehyde;

10-methoxy camptothecin-7-aldehyde;

7-acetoxymethyl camptothecin;

7-acetoxym ethyl- 10-methyl camptothecin;

7-cyano-lO-methoxy camptothecin;

7-cyano camptothecin;

7-formylethenyl camptothecin;

7-ethoxycarbonylethenyl camptothecin;

7-cyanoethenyl camptothecin;

7-(2,2-dicyanoethenyl) camptothecin;

7-(2-cyano-2-ethoxycarbonyl)ethenyl camptothecin;

7-ethoxycarbonylethyl camptothecin;

7-ethyl camptothecin;

7-n-propyl camptothecin;

7-acetoxymethyl camptothecin;

7-n-propylcarbonyloxymethyl camptothecin ;

7-ethoxycarbonyl camptothecin;

7-ethyl- 10-hydroxy camptothecin;

7-ethyl- 10-acetyloxy camptothecin;

7-methyl- 10-aminocarbonyloxy camptothecin ;

7-n-propyl- 10-piperidino cazbonyloxy camptothecin;

7-ethyl- 10-(2-dimethylamino)ethyl camptothecin;

7-ethyl- 10-[4(l-piperidino)-piperidino carbonyloxy camptothecin;

7-ethyl- 10-(l -piperazine)carbonyloxy camptothecin; 7-ethyl- 10-(4-i-propylaminocarbonylmethylpiperazine)carbonyloxy camptothecin;

7-ethyl-10-[4(l-pyrrolidinyl)piperazine]carbonyloxy camptothecin; 7-ethyl- 10-[(4-(dimethylamino)-l -piperidino]carbonyloxy camptothecin;

7-ethyl- 10-[4-(di-n-propylamino)- 1 -piperidinol]carbonyloxy camptothecin;

/ 7-ethyl-10-[(4-(di-n-butylamino)-l-piperidino]carbonyloxy camptothecin;

7-ethyl- 10-[4-(l-pyrrolidino)-l-piperidino)]carbonyloxy camptothecin;

7-ethyl- 10-[4-(l -piperidino)-l -piperidino]carbonyloxy camptothecin; and

7-ethyl- 10-[N-methyl-N-2-(dimethylamino)ethyl amino]carbonyloxy camptothecin; or a pharmaceutically acceptable salt or prodrug thereof, wherein said compound has 20(S) or 20(RS) configuration.

24. The pharmaceutical composition of claim 22, further comprising one or more active agents independently selected from the group consisting of chemotherapeutic agents, anti-angiogenesis agents, vascular targeting agents, HEFl inhibitors, Hsp90 inhibitors, a tyrosine kinase inhibitor, a serine/threonine kinase inhibitor, a proteasome inhibitor, an HDAC inibitor, a caspase inducer, a CDK inhibitor, and a proapoptotic molecule.

25. The pharmaceutical composition of claim 24, wherein said chemotherapeutic agent is selected from the group consisting of abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic trioxide, asparaginase, BCG live, bevaceizumab, bexarotene, bleomycin, bortezomib, busulfan, calusterone, camptothecin, capecitabine, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cinacalcet, cisplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, darbepoetin alfa, daunorubicin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone, Elliott's B solution, epirubicin, epoetin alfa, estramustine, etoposide, exemestane, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gemcitabine, gerntuzumab ozogamicin, gefitmib, goserelin, hydroxyurea, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib, interferon alfa-2a, interferon alfa-2b, irinotecan, letrozole, leucovorin, levamisole, lomustine, meclorethamine, megestrol, melphalan, mercaptopurine, mesna, methotrexate, methoxsalen, methylprednisolone, mitomycin C, mitotane, mitoxantrone, nandrolone, nofetumomab, oblimersen, oprelvekin, oxaliplatin, paclitaxel, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed, pentostatin, pipobroman, plicamycin, polifeprosan, porfimer, procarbazine, quinacrine, rasburicase, rituximab, sargramostim, streptozocin, talc, tamoxifen, tarceva, temozolomide, teniposide, testolactone, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, and zoledronate.

26. The pharmaceutical composition of claim 24, wherein said anti-angiogenesis agent is selected from the group consisting of bevacizumab, angiostatin, endostatin, batimastat, captopril, cartilage derived inhibitor, genistein, interleukin 12, lavendustin, medroxypregesterone acetate, recombinant human platelet factor 4, tecogalan, thrombospondin, TNP-470, anti-VEGF monoclonal antibody, soluble VEGF-receptor chimaeric protein, anti-VEGF receptor antibodies, anti-PDGF receptors, inhibitors of integrins, tyrosine kinase inhibitors, serine/threonine kinase inhibitors, antisense oligonucleotides, antisense oligodexoynucleotides, siRNAs, anti-VEGF ap tamers and pigment epithelium derived factor.