WO2003075828A2

WO2003075828A2 - Compounds useful in the treatment of cancer

Info

Publication number: WO2003075828A2
Application number: PCT/IL2003/000198
Authority: WO
Inventors: Dan Jacob Gelvan; Lev Goltsman; Alexander Chausovsky
Original assignee: Zetiq Technologies Ltd.
Priority date: 2002-03-11
Filing date: 2003-03-11
Publication date: 2003-09-18
Also published as: WO2003075828A3; AU2003212634A1; AU2003212634A8

Abstract

The present invention provides a) a method of inhibiting the growth of a cancer cell; b) a method of inhibiting cell proliferation; and c) a method of inducing cell death. The methods comprise treating the cell with a compound represented by any of the structures of formulas I-XV, as defined herein. The present invention further provides to a) a method of treating a subject having cancer; and b) a method of delaying the progression of cancer in a subject.

Description

COMPOUNDS USEFUL IN THE TREATMENT OF CANCER

FIELD OF INVENTION

[0001] The present invention relates to compounds, compositions and methods of use for the treatment of cancer. More specifically, the present invention relates to compounds, which specifically inhibit the growth of cancer cells, and to the use of these compounds and pharmaceutical compositions comprising these compounds for the treatment of cancer.

BACKGROUND OF THE INVENTION

[0002] Cancer is a disorder in which a population of cells has become, in varying degrees, unresponsive to the control mechanisms that normally govern proliferation and differentiation. The leading therapies to date are surgery, radiation and chemotherapy.

[0003] Traditionally, chemotherapeutic treatment of cancer has focused on killing cancer cells directly by exposing them to cytotoxic substances. Ideally cytotoxic agents are specific for cancer and tumor cells while not affecting, or having a mild effect on normal cells. Unfortunately, most cytotoxic agents target especially rapidly dividing cells (both tumor and normal) and thus injure both neoplastic and normal cell populations.

[0004] There is an urgent and ongoing need to develop new therapeutic approaches to the treatment of cancer, particularly chemical compounds which are easily obtainable, and which inhibit the growth/proliferation of cancer tissues, while having little or no effect on normal tissue.

SUMMARY OF THE INVENTION

[0005] In one embodiment, the present invention provides a method of inhibiting the growth of a cancer cell, comprising the step of contacting the cell with a compound represented by the structure of formula I, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to inhibit the growth of the cancer cell

wherein Ri is H, a Cι-C₄ linear or branched alkyl or — N _ wherein

Rs'

R₅ and R₅' are independently of each other H, a Cι-C linear or branched alkyl,

_/=v^R- or R₅ and R₅' together with the nitrogen to which they are attached represent //

a group of the .

R₆ is H, a Cι-C linear or branched alkyl, hydroxy, alkoxy, halogen, carboxy or

COORu, wherein

Ru is a C₁-C4 linear or branched alkyl;

R₇ and R₇' are independently of each other H, a Cι-C linear or branched alkyl, hydroxy, alkoxy, halogen or carboxy; R₂ and R₃ are independently of each other H, a C1-C4 linear or branched alkyl, — N .

R₈'

— OR₉, , wherein

R₈ and R₈' are independently of each other H, O, a Cι-C linear or branched alkyl, / , -(CH₂) _nOR, wherein n is an integer of 1-4 and R is H or a C1-C₄

linear or branched alkyl, or R₈ and R₈' together with the nitrogen to which they are attached represent a group of the formula:

R is H, a C₁-C4 linear or branched alkyl, or / K ;

Rio and Rio' are independently of each other H, a C1-C4 linear or branched alkyl, or — /^~ ;

and

R₄ is H, a C₁-C4 linear or branched alkyl, phenyl or NO₂.

[0006] In another embodiment, the present invention provides a method of inducing cell death, comprising the step of contacting a cell with a compound represented by the structure of formula I, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to induce the death of said cell.

I wherein

Ri is H, a C1-C₄ linear or branched alkyl or — N , wherein

R₅'

R₅ and R₅' are independently of each other H, a C₁-C₄ linear or branched alkyl, / K , or R₅ and R5' together with the nitrogen to which they are

attached represent a group .

R_δ is H, a Cι-C₄ linear or branched alkyl, hydroxy, alkoxy, halogen, carboxy or COOR₁₁ wherein Ru is a C₁-C4 linear or branched alkyl;

R and R ' are independently of each other H, a C₁-C₄ linear or branched alkyl, hydroxy, alkoxy, halogen or carboxy;

R₂ and R₃ are independently of each other H, a Cι-C linear or branched alkyl,

-N rein

R₈ and R₈' are independently of each other H, O, a C₁-C₄ linear or branched alkyl, / K , -(CH₂) _nOR wherein n is an integer of 1-4 and R is H or a Cι-C₄ linear

or branched alkyl, or R₈ and R₈' together with the nitrogen to which they are attached represent a group of the formula

R₉ is H, a C₁-C₄ linear or branched alkyl, or_/ ;

Rio and Rio' are independently of each other H, a C1-C₄ linear or branched alkyl, or /^{^}X^;

and R4 is H, a C₁-C₄ linear or branched alkyl, phenyl or NO₂.

[0007] In another embodiment, the present invention provides a method of treating a subject having cancer, comprising the step of administering to said subject a compound represented by the structure of formula I, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to treat cancer in said subject.

I wherein 5 Ri is H, a C₁-C₄ linear or branched alkyl or — N , wherein

R₅'

R₅ and R5' are independently of each other H, a C1-C₄ linear or branched alkyl, or R₅ and R₅' together with the nitrogen to which they are attached

represent a group -

Rs is H, a Cι-C₄ linear or branched alkyl, hydroxy, alkoxy, halogen, carboxy or COOR1₁, wherein Ru is a Cι-C₄ linear or branched alkyl;

R and R₇' are independently of each other H, a Cι-C₄ linear or branched alkyl, hydroxy, alkoxy, halogen or carboxy; R₂ and R₃ are independently of each other H, a C1-C4 linear or branched alkyl,

— , wherein

R₈ and R₈' are independently of each other H, O, a C₁-C₄ linear or branched alkyl, / /=y^Rβ , -(CH₂) _nOR wherein n is an integer of 1-4 and R is H or a C₁-C₄ linear

R₉ is H, a C1-C4 linear or branched alkyl, or / K ;

are independently of each other H, a Cι-C linear or branched alkyl,

and R-₄ is H, a C₁-C₄ linear or branched alkyl, phenyl or NO₂.

[0008] In another embodiment, the present invention provides a method of delaying the progression of cancer in a subject having cancer, comprising the step of administering to said subject a compound represented by the structure of formula I, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to delay the progression of cancer in said subject

wherein

Ri is H, a C₁-C₄ linear or branched alkyl

R₅ and R₅' are independently of each other H, a Cι-C₄ linear or branched alkyl, / K , or R₅ and R₅' together with the nitrogen to which they are

₅ is H, a C₁-C₄ linear or branched alkyl, hydroxy, alkoxy, halogen, carboxy or

COOR1₁, wherein Ru is a C1-C4 linear or branched alkyl;

R and R₇' are independently of each other H, a C₁-C₄ linear or branched alkyl, hydroxy, alkoxy, halogen or carboxy; linear or branched alkyl,

R₈ and R₈' are independently of each other H, O, a C1-C₄ linear or branched alkyl, / K ,-(CH₂) „OR wherein n is an integer of 1-4 and R is H or a C1-C₄ linear

=\,Rs

R is H, a C₁-C₄ linear or branched alkyl, or.

^■- ^;

Rio and R₁₀' are independently of each other H, a C₁-C4 linear or branched alkyl, or — ^~~\ ⁶;

and

R₄ is H, a C₁-C4 linear or branched alkyl, phenyl or NO₂.

[0009] In another embodiment, the present invention provides a method of inhibiting the growth of a cancer cell, comprising the step of contacting said cell with a compound represented by the structure of formula XII, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to inhibit the growth of said cancer cell.

xπ wherein

Ri is a C₁-C₄ linear or branched alkyl, or Ri is

R₂ and R₃ are independently of each other H, a Cι-C₄ linear or branched alkyl, a C₁-C₄ linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, COR, COOR, COOH, NHCOR, CF₃, NO₂, or R₂ and R₃ together with the benzene group to which they are attached represent a fused ring system represented by the structure:

R4 and R₅ are independently of each other H, a Cι-C₄ linear or branched alkyl, a C₁-C₄ linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, CF₃ or NO₂;

R-s is H or a C1-C4 linear or branched alkyl;

R₇ and R₈ are independently of each other H, a Cι-C₄ linear or branched alkyl, halogen, hydroxy, alkoxy, thio, alkylthio, CF₃, or O₂; and R is H or a C1-C₄ linear or branched alkyl.

[00010] In another embodiment, the present invention provides a method of inducing cell death, comprising the step of contacting a cell with a compound represented by the structure of formula XII, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to induce the death of said cell.

xπ wherein

Ri is a Cι-C linear or branched alkyl, or Ri is

R₂ and R₃ are independently of each other H, a C₁-C₄ linear or branched alkyl, a C₁-C₄ linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, COR, COOR, COOH, NHCOR, CF₃, NO₂, or R₂ and R₃ together with the benzene group to which they are attached represent a fused ring system represented by the structure:

R₄ and R₅ are independently of each other H, a C1-C4 linear or branched alkyl, a Cι-C linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, CF₃, or NO₂;

Rδ is H or a C1-C4 linear or branched alkyl;

R₇ and R₈ are independently of each other H, a Cι-C linear or branched alkyl, halogen, hydroxy, alkoxy, thio, alkylthio, CF₃, or NO₂; and

R is H or a C1-C4 linear or branched alkyl.

[00011] In another embodiment, the present invention provides a method of treating a subject having cancer, comprising the step of administering to said subject a compound represented by the structure of formula XII, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to treat cancer in said subject

XII wherein

Ri is a C₁-C₄ linear or branched alkyl, or Ri

R₂ and R₃ are independently of each other H, a C₁-C4 linear or branched alkyl, a C₁-C₄ linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, COR, COOR, COOH, NHCOR, CF₃, NO₂, or R₂ and R₃ together with the benzene group to which they are attached represent a fused ring system represented by the structure:

R4 and R5 are independently of each other H, a C₁-C₄ linear or branched alkyl, a C1-C4 linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, CF₃ or NO₂;

Rδ is H or a Cι-C linear or branched alkyl;

R₇ and R₈ are independently of each other H, a C₁-C₄ linear or branched alkyl, halogen, hydroxy, alkoxy, thio, alkylthio, CF₃, or NO₂; and

R is H or a C₁-C4 linear or branched alkyl.

[00012] In another embodiment, the present invention provides a method of delaying the progression of cancer in a subject having cancer, comprising the step of administering to said subject a compound represented by the structure of formula XII or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to delay the progression of cancer in said subject.

XII wherein

Ri is a C₁-C₄ linear or branched alkyl, or Ri is

R₂ and R₃ are independently of each other H, a C1-C4 linear or branched alkyl, a C₁-C4 linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, COR, COOR, COOH, NHCOR, CF₃, NO₂, or R₂ and R₃ together with the benzene group to which they are attached represent a fused ring system represented by the structure:

R₄ and R₅ are independently of each other H, a C₁-C₄ linear or branched alkyl, a C₁-C₄ linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, CF₃ or NO₂;

Rg is H or a C₁-C₄ linear or branched alkyl;

R and R₈ are independently of each other H, a C₁-C₄ linear or branched alkyl, halogen, hydroxy, alkoxy, thio, alkylthio, CF₃, or NO₂; and

R is H or a C₁-C₄ linear or branched alkyl.

BRIEF DESCRIPTION OF THE FIGURES

[00013] The present invention will be understood and appreciated more fully from the following detailed description taken in conjunction with the appended drawings in which:

Figure 1. Structures, chemical formulas and molecular weights for ZTQIOOI - ZTQ1008.

Figure 2 A. Effect of increased concentration of ZTQIOOI on the optical density at 630 nm of HT-29, WiDr, and CCD-33Co cells stained with Methylene Blue.

Figure 2B. Effect of increased concentration of ZTQ1002 on the optical density at 630 nm of HT-29, WiDr, and CCD-33Co cells stained with Methylene

Blue.

Figure 2C. Effect of increased concentration of ZTQ1003 on the optical density at 630 nm of HT-29, WiDr, and CCD-33Co cells stained with Methylene Blue.

Figure 2D. Effect of increased concentration of ZTQ1004 on the optical density at 630 nm of HT-29, WiDr, and CCD-33Co cells stained with Methylene Blue.

Figure 2E. Effect of increased concentration of ZTQ1005 on the optical density at 630 nm of HT-29, WiDr, and CCD-33Co cells stained with Methylene Blue. Figure 3 A. Effect of increased concentration of ZTQIOOI respectively ZTQ1002 on the growth of highly proliferative HT-29 cells, and on non-proliferating CCD-33Co cells.

Figure 3B. Effect of increased concentration of ZTQ1003 respectively ZTQ1004 on the growth of highly proliferative HT-29 cells, and on non-proliferating

CCD-33Co cells.

Figure 3C. Effect of increased concentration of ZTQ1005 respectively ZTQ1006 on the growth of highly proliferative HT-29 cells, and on non-proliferating CCD-33Co cells.

Figure 3D. Effect of increased concentration of ZTQ1007 respectively ZTQ1008 on the growth of highly proliferative HT-29 cells, and on non-proliferating CCD-33Co cells. Figure 4. GI₅₀ values (the drug concentration resulting in a 50% reduction in the net protein increase relative to the net protein increase in untreated cells). NT = not tested. Empty cells indicate a GI50 value higher than 30 μM.

Figure 5. Effect of ZTQ1001-ZTQ1005 on the growth of HT-29 cells under non- solid support conditions. The treatments were carried out at the indicated concentrations and the cells were grown in soft agar for 13 days after treatment. *Colonies counted after 10 days of growth in a separate experiment. ⁺Colonies counted after 11 days of growth in a separate experiment.

Figure 6. Effect of treatment on hemoglobin production in K-562 cells. The cells were treated for 72 h with 0.1 μM ZTQIOOI, 0.5 μM ZTQIOOI or Gleevec™, respectively, followed by retraction for 174 h. Untreated cells were used as negative control.

Figure 7A. Formazan deposits in non-treated U-937 human leukemia cells. Figure 7B. Formazan deposits in U-937 human leukemia cells treated with 300 nM

ZTQIOOI.

Figure 7C. Formazan deposits in U-937 human leukemia cells treated with 0.8 mM sodium butyrate.

Figure 8 A. Untreated MCF-7 cells stained with Nile Red, lipid stain.

Figure 8B. MCF-7 cells treated with 2μM ZTQIOOI and stained with Nile Red, lipid stain.

Figure 8C. MCF-7 cells treated with 2.5 mM sodium butyrate and stained with

Nile Red, lipid stain.

Figure 9 A. DNA stained MCF-7 cells treated with 1 μM ZTQ 1001 for 72 h.

Figure 9B. DNA stained MCF-7 cells treated with 2.5 mM sodium butyrate for 72 h.

Figure 9C. DNA stained untreated MCF-7 cells.

Figure 10 A. Microtubule network visualization by immunofluorescence staining with anti-α-tubulin antibodies of MCF-7 cells treated with 1 μM ZTQIOOI for 72 h. Figure 10B. Microtubule network visualization by immunofluorescence staining with anti-α-tubulin antibodies of MCF-7 cells treated with 2.5 mM sodium butyrate for 72 h. Figure IOC Microtubule network visualization by immunofluorescence staining with anti-α-tubulin antibodies of untreated MCF-7 cells

Figure 11 A Microtubule network visualization by immunofluorescence staining with anti-α-tubulin antibodies of MCF-7 cells treated with 2μM ZTQIOOI for

30 min

Figure 11B Microtubule network visualization by immunofluorescence staining with anti-α-tubulin antibodies of untreated MCF-7 cells

Figure 12 Effect of ZTQIOOI, Nincristine and Taxol on the polymerization of purified tubulin

Figure 13 Effect of increasing concentrations of ZTQIOOI and ZTQ 1005 on the de-polymerization of Taxol stabilized tubulin

Figure 14 Effect of ex- vivo treatment of HT-29* cells with ZTQ 1003 on tumor growth after subcutaneous implantation of treated cells into nude mice

Cells treated with Genistein and untreated cells were used for positive and negative controls, respectively n = 7-10 mice per group Mean ± SE

For each treatment the tumor volume was significant different from the untreated control, P < 0 05, t-test

Figure 15 Effect of ex- vivo treatment of HT-29 * cells with ZTQ 1002 on tumor growth after subcutaneous implantation of treated cells into nude mice

Cells treated with Genistein and untreated cells were used for positive and negative controls, respectively n = 7-10 mice per group Mean ± SE For each treatment the tumor volume was significant different from the untreated control, P < 0 05, t-test

DETAILED DESCRIPTION OF THE INVENTION

[00014] The present invention provides a) a method of inhibiting the growth of a cancer cell; b) a method of inhibiting cell proliferation; and c) a method of inducing cell death. The methods comprise treating the cell with a compound represented by any of the structures of formulas I-XV, as defined herein. The present invention further provides to a) a method of treating a subject having cancer; and b) a method of delaying the progression of cancer in a subject. The methods comprise administering to the subject a compound represented by any of the structures of formulas I-XV, as defined herein.

[000151 In one embodiment, the present invention provides a compound represented by any of the structures of formulas I-XV.

[00016] In one embodiment of the present invention, the compound which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula I. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula I. In another embodiment, the compound is a hydrate of the compound of formula I. In another embodiment, the compound is a combination of any of compound I, its pharmaceutically acceptable salt and/or hydrate thereof.

[00017] In one embodiment of the present invention, the compound which is useful in selectively inhibiting the growth of a cancer cell, in selectively inhibiting cell proliferation, in selectively inducing cell death, is a compound of formula I.

I wherein Ri is H, a C₁-C₄ linear or branched alkyl

R₅ and R5' are independently of each other H, a C1-C₄ linear or branched alkyl, / K , or R₅ and R5' together with the nitrogen to which they are

attached represent a group of the or — N b .

Rδ is H, a C₁-C₄ linear or branched alkyl, hydroxy, alkoxy, halogen, carboxy or COOR11, wherein Ru is a C₁-C₄ linear or branched alkyl;

R₇ and R₇' are independently of each other H, a C1-C4 linear or branched alkyl, hydroxy, alkoxy, halogen or carboxy; H, a C1-C4 linear or branched alkyl, wherein

Rs and R₈' are independently of each other H, O, a C1-C₄ linear or branched alkyl,_/ , -(CH₂) _nOR, wherein n is an integer of 1-4 and R is H or a C₁-C₄ linear

R9 is H, a C₁-C₄ linear or branched alkyl, or / K ;

Rio and Rio' are independently of each other H, a C₁-C₄ linear or branched alkyl, or

and

R₄ is H, a C₁-C₄ linear or branched alkyl, phenyl or NO₂.

[00018] In one embodiment of the present invention, Ri in compound I is CH₃.

[00019] In another embodiment, Ri is NH₂.

[00020] In another embodiment, Ri in compound I is

[00021] In another embodiment, Ri in compound I is

[00022] In one embodiment of the present invention, R₂ in compound I is CH₃.

[00023] In another embodiment, R₂ is

[00024] In another embodiment, R₂ is —

[00025] In another embodiment, R₂ in compound I

[00026] In another embodiment, R₂ in compound I

[00027] In one embodiment of the present invention, R₃ in compound I is phenyl. In another embodiment, R₃ in compound I is — OR₉, wherein R₉ is / K .

O II

[00028] In another embodiment, R₃ in compound I is — o — — C— OCH₃.

[00029] In another embodiment, R₃ in compound I is ^■ — NR₈R₈\ wherein one of R₈ and R₈' is H and the other is

[00030] n compound I is

[00031] In another embodiment, R₃

[00032] In another embodiment, R₃ in compound I

[00033] In another embodiment, the compound, which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula II. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula II. In another embodiment, the compound is a hydrate of the compound of formula II. In another embodiment, the compound is a combination of any of compound

II, its pharmaceutically acceptable salt and/or hydrate thereof.

[00034] In one embodiment of the present invention, R₇ and R₇- in compound II are both CH₃.

[00035] In another embodiment, one of Rio and Rio' in compound II is CH₃ and the other is 4-methylphenyl.

[00036] In another embodiment, the compound which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula III. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula III. In another embodiment, the compound is a hydrate of the compound of formula III. In another embodiment, the compound is a combination of any of compound

III, its pharmaceutically acceptable salt and/or hydrate thereof.

[00037] In another embodiment, the compound which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula IN. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula IN. In another embodiment, the compound is a hydrate of the compound of formula IN. In another embodiment, the compound is a combination of any of compound IN, its pharmaceutically acceptable salt and/or hydrate thereof.

[00038] In one embodiment of the present invention, Rδ in compound IV is H. In another embodiment, the compound which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula N. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula N. In another embodiment, the compound is a hydrate of the compound of formula N. In another embodiment, the compound is a combination of any of compound N, its pharmaceutically acceptable salt and/or hydrate thereof.

[00039] In another embodiment, the compound which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula VI. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula VI. In another embodiment, the compound is a hydrate of the compound of formula VI. In another embodiment, the compound is a combination of any of compound VI, its pharmaceutically acceptable salt and/or hydrate thereof.

[00040] In one embodiment of the present invention, R-_δ in compound VI is COOCH₃. In another embodiment, R and Rr in compound VI are both CH₃. In another embodiment, the compound which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula VII. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula NIL In another embodiment, the compound is a hydrate of the compound of formula NIL In another embodiment, the compound is a combination of any of compound Nil, its pharmaceutically acceptable salt and/or hydrate thereof.

[00041] In another embodiment, the compound, which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula NIII. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula VIII. In another embodiment, the compound is a hydrate of the compound of formula NIII. In another embodiment, the compound is a combination of any of compound NIII, its pharmaceutically acceptable salt and/or hydrate thereof.

[00042] In one embodiment of the present invention, R_δ in compound VIII is H. In another embodiment, the compound, which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula IX. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula IX. In another embodiment, the compound is a hydrate of the compound of formula IX. In another embodiment, the compound is a combination of any of compound IX, its pharmaceutically acceptable salt and/or hydrate thereof.

[00043] In another embodiment, the compound, which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula X. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula X. In another embodiment, the compound is a hydrate of the compound of formula X In another embodiment, the compound is a combination of any of compound X, its pharmaceutically acceptable salt and/or hydrate thereof.

X R ^N ₇7

[00044] In one embodiment of the present invention, Rg in compound X is OCH₃. In another embodiment, R₇ and R - in compound X are both CH₃. In another embodiment, the compound, which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula XI. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula XI. In another embodiment, the compound is a hydrate of the compound of formula XI. In another embodiment, the compound is a combination of any of compound XI, its pharmaceutically acceptable salt and/or hydrate thereof.

[00045] In another embodiment, the compound, which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula XII. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula XII. In another embodiment, the compound is a hydrate of the compound of formula XII. In another embodiment, the compound is a combination of any of compound XII, its pharmaceutically acceptable salt and/or hydrate thereof.

[00046] In one embodiment of the present invention, the compound which is useful in selectively inhibiting the growth of a cancer cell, in selectively inhibiting cell proliferation, in selectively inducing cell death, is a compound of formula XII.

xn wherein

Ri is a C₁-C4 linear or branched alkyl, or Ri

R₂ and R₃ are independently of each other H, a C₁-C4 linear or branched alkyl, a Cι-C linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, COR, COOR, COOH, NHCOR, CF₃, NO₂, or R₂ and R₃ together with the benzene group to which they are attached represent a fused ring system represented by the structure:

R₄ and R₅ are independently of each other H, a C₁-C₄ linear or branched alkyl, a C₁-C4 linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, CF₃ or NO₂;

Rδ is H or a C1-C₄ linear or branched alkyl;

R₇ and R₈ are independently of each other H, a C1-C4 linear or branched alkyl, halogen, hydroxy, alkoxy, thio, alkylthio, CF₃, or NO₂; and

R is H or a C₁-C₄ linear or branched alkyl. [00047] In one embodiment of the present invention, Ri is CH₃. In another embodiment, Ri is phenyl. In another embodiment, R₂ is ethoxy and R₃ is H. In another embodiment, R₂ is bromo and R₃ is H. In another embodiment, R₂ is chloro and R₃ is H. In another embodiment, R4 is methoxy and R₅ is H. In another embodiment, R₄ is ethoxy and R5 is H.

[00048] In another embodiment, the compound, which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula XIII. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula XIII. In another embodiment, the compound is a hydrate of the compound of formula XIII. In another embodiment, the compound is a combination of any of compound XIII, its pharmaceutically acceptable salt and/or hydrate thereof.

[00049] In another embodiment, the compound, which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula XIV. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula XIV. In another embodiment, the compound is a hydrate of the compound of formula XIV. In another embodiment, the compound is a combination of any of compound XIV, its pharmaceutically acceptable salt and/or hydrate thereof.

[00050] In another embodiment, the compound, which is useful in inhibiting the growth of a cancer cell, in inhibiting cell proliferation, in inducing cell death, in treating cancer and in delaying the progression of cancer, is a compound of formula XV. In another embodiment, the compound is a pharmaceutically acceptable salt of the compound of formula XV. In another embodiment, the compound is a hydrate of the compound of formula XV. In another embodiment, the compound is a combination of any of compound XV, its pharmaceutically acceptable salt and/or hydrate thereof.

[00051] The following definitions are used in the present invention:

[00052] An "alkyl" group refers to a saturated aliphatic hydrocarbon, including straight-chain, branched-chain and cyclic alkyl groups. In one embodiment of the present invention, the alkyl group has 1-4 carbons. In another embodiment, the alkyl group has 1 carbon (methyl). In another embodiment, the alkyl group has 2 carbons (ethyl). In another embodiment, the alkyl group has 3 carbons (such as propyl or isopropyl). In another embodiment, the alkyl group has 4 carbons (such as butyl, isobutyl, sec-butyl and tert-butyl). The alkyl group may be unsubstituted or substituted by one or more groups selected from halogen, hydroxy, alkoxy carbonyl, amido, alkylamido, dialkylamido, nitro, amino, alkylamino, dialkylamino, carboxyl, haloalkyl, aryl, thio and thio alkyl.

[00053] A "haloalkyl" group refers to an alkyl group as defined above, which is substituted by one or more halogen atoms, e.g. by F, CI, Br or I.

[00054] An "aryl" group refers to an aromatic group having at least one carbocyclic aromatic group or heterocyclic aromatic group, which may be unsubstituted or substituted by one or more groups selected from halogen, haloalkyl, hydroxy, alkoxy carbonyl, amido, alkylamido, dialkylamido, nitro, amino, alkylamino, dialkylamino, carboxy or thio or thioalkyl. Nonlimiting examples of aryl rings are phenyl, naphthyl, pyranyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyrazolyl, pyridinyl, furanyl, thiophenyl, thiazolyl, imidazolyl, isoxazolyl, and the like.

[00055] A "hydroxyl" group refers to an OH group. An "alkoxy" group refers to an O-alkyl group, wherein alkyl has the same definition as described above. A "phenoxy" group refers to an O-phenyl group. A "thio" group refers to an SH group.

[00056] An "alkylthio" group refers to an S-aryl group wherein aryl has the same definition as described above. [00057] A halogen or halo group refers to F, CI, Br or I.

[00058] As contemplated herein, the present invention relates to the use of any one of compounds of formulas I-XV and/or their pharmaceutically acceptable salts or hydrates for preparing a medicament for inhibiting the growth of a cancer cell, inhibiting cell proliferation, inducing cell death, treating a subject having cancer and/or delaying the progression of cancer in a subject. In another embodiment, the present invention relates to the use of an analog, derivative, isomer, metabolite, N-oxide or any combination thereof of any of the compounds of formulas I-XV. In one embodiment, the invention relates to the use of an analog of a compound according to any of formulas I-XV. In another embodiment, the invention relates to the use of a derivative of a compound according to any of formulas I-XV. In another embodiment, the invention relates to the use of an isomer of a compound according to any of formulas I-XV. In another embodiment, the invention relates to the use of a metabolite of a compound according to any of formulas I-XV. In another embodiment, the invention relates to the use of a pharmaceutically acceptable salt of a compound according to any of formulas I-XV. In another embodiment, the invention relates to the use of a hydrate of a compound according to any of formulas I-XV. In another embodiment, the invention relates to the use of an N-oxide of a compound according to any of formulas I-XV.

[00059] As defined herein, the term "isomer" includes, but is not limited to, optical isomers and analogs, structural isomers and analogs, conformational isomers and analogs, and the like.

[00060] In one embodiment, this invention encompasses the use of various optical isomers of the compounds of the present invention. It will be appreciated by those skilled in the art that the compounds of the present invention may contain at least one chiral center. Accordingly, the compounds used in the methods of the present invention may exist in, and be isolated in, optically-active or racemic forms. Some compounds may also exhibit polymorphism. It is to be understood that the present invention encompasses any racemic, optically-active, polymorphic, or stereroisomeric form, or mixtures thereof, which form possesses properties useful in the treatment of cancer-related conditions described herein. In one embodiment of the present invention, the compounds are the pure (R)-isomers. In another embodiment, the compounds are the pure (S)-isomers. In another embodiment, the compounds are a mixture of the (R) and the (S) isomers. In another embodiment, the compounds are a racemic mixture comprising an equal amount of the (R) and the (S) isomers. It is well known in the art how to prepare optically-active forms (for example, by resolution of the racemic form by recrystallization techniques, by synthesis from optically-active starting materials, by chiral synthesis, or by chromatographic separation using a chiral stationary phase).

[00061] In one embodiment, this invention encompasses the use of various structural isomers of the compounds of the present invention. It will be appreciated by those skilled in the art that the compounds of the present invention may exist as the (Z)- or the (E)-isomers. The invention encompasses pure (Z)- and (E)- isomers of the compounds defined herein and mixtures thereof.

[00062] The invention includes pharmaceutically acceptable salts of amino-substituted compounds with organic and inorganic acids, for example, citric acid and hydrochloric acid. The invention also includes N-oxides of the amino substituents of the compounds described herein. Pharmaceutically acceptable salts can also be prepared from the phenolic compounds by treatment with inorganic bases, for example, sodium hydroxide. Also, esters of the phenolic compounds can be made with aliphatic and aromatic carboxylic acids, for example, acetic acid and benzoic acid esters.

[00063] This invention further includes derivatives of the compounds of the present invention. The term "derivatives" includes but is not limited to ether derivatives, acid derivatives, amide derivatives, ester derivatives and the like. In addition, this invention further includes hydrates of the compounds of the present invention. The term "hydrate" includes but is not limited to hemihydrate, monohydrate, dihydrate, trihydrate and the like.

[00064] This invention further includes metabolites of the compounds of the present invention compounds. The term "metabolite" means any substance produced from another substance by metabolism or a metabolic process.

[00065] Thus, in accordance with one embodiment of the present invention, there is provided a method of inhibiting the growth of a cancer cell, comprising the step of contacting the cell with a compound represented by the structure of formula I as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to inhibit the growth of the cancer cell. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula FI as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula III as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula IV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula V as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula VI as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula VTI as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula VIII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula IX as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula X as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XI as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XIII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XIV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof

[00066] Furthermore, in accordance with another embodiment of the present invention, there is provided a method of inhibiting cell proliferation, comprising the step of contacting a cell with a compound represented by the structure of formula I as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to inhibit proliferation of the cell. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula II as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula III as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula IV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula V as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula VI as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula VII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula VIII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula IX as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula X as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XI as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XIII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XIV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof.

[00067] Furthermore, in accordance with another embodiment of the present invention, there is provided a method of inducing cell death, comprising the step of contacting a cell with a compound represented by the structure of formula I as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to induce the death of the cell. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula II as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula III as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula IV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula V as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula VI as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula VII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula VIII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula IX as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula X as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XI as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XIII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XIV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises contacting the cell with a compound represented by the structure of formula XV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. [00068] Furthermore, in accordance with another embodiment of the present invention, there is provided a method of treating a subject having cancer, comprising the step of administering to the subject a compound represented by the structure of formula I as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to treat cancer in the subject. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula II as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula III as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula IV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula V as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula VI as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula VII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula VIII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula IX as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula X as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula XI as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula XII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula XIII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula XIV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula XV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof.

[00069] Furthermore, in accordance with another embodiment of the present invention, there is provided a method of delaying the progression of cancer in a subject having cancer, comprising the step of administering to said subject a compound represented by the structure of formula I as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to delay the progression of cancer in the subject. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula II as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula III as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula IV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula V as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula VI as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula VII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula VIII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula IX as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula X as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula Xt as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula XII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula XIII as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula XIV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof. In another embodiment, the method comprises administering to the subject a compound represented by the structure of formula XV as defined hereinabove, or its pharmaceutically acceptable salt, hydrate or any combination thereof.

[00070] In one embodiment of the present invention, the cell is a cancer cell. In another embodiment, the cancer cell is a colon cancer cell. In another embodiment, the cancer is colon cancer.

[00071] A "cancer cell" is defined herein as a neoplastic cell, a pre-malignant cell, a metastatic cell, a malignant cell, a tumor cell, an oncogenic cell, a cell with a cancer genotype, a cell of malignant phenotype, a cell with a malignant genotype, a cell displaying cancer associated metabolic atypia, an oncogene transfected cell, a virus transformed cell, a cell which expresses a marker for an oncogene, a cell which expresses a marker for cancer, or a combination thereof.

[00072] In accordance alternative embodiments of the present invention, malignant cell is an adenocarcinoma cell, an adrenal gland tumor cell, an ameloblastoma cell, an anaplastic cell, anaplastic carcinoma of the thyroid cell, an angiofibroma cell, an angioma cell, an angiosarcoma cell, an apudoma cell, an argentaffmoma cell, an arrhenoblastoma cell, an ascites tumor cell, an ascitic tumor cell, an astroblastoma cell, an astrocytoma cell, an ataxia-telangiectasia cell, an atrial myxoma cell, a basal cell carcinoma cell, a benign tumor cell, a bone cancer cell, a bone tumor cell, a brainstem glioma cell, a brain tumor cell, a breast cancer cell, a Burkitt's lymphoma cell, a cancerous cell, a carcinoid cell, a carcinoma cell, a cerebellar astrocytoma cell, a cervical cancer cell, a cherry angioma cell, a cholangiocarcinoma cell, a cholangioma cell, a chondroblastoma cell, a chondroma cell, a chondro sarcoma cell, a chorioblastoma cell, a choriocarcinoma cell, a colon cancer cell, a common acute lymphoblastic leukaemia cell, a craniopharyngioma cell, a cystocarcinoma cell, a cystofibroma cell, a cystoma cell, a cytoma cell, a ductal carcinoma in situ cell, a ductal papilloma cell, a dysgerminoma cell, an encephaloma cell, an endometrial carcinoma cell, an endothelioma cell, an ependymoma cell, an epithelioma cell, an erythroleukaemia cell, an Ewing's sarcoma cell, an extra nodal lymphoma cell, a feline sarcoma cell, a fibroadenoma cell, a fibrosarcoma cell, a follicular cancer of the thyroid cell, a ganglioglioma cell, a gastrinoma cell, a glioblastoma multiforme cell, a glioma cell, a gonadoblastoma cell, an haemangioblastoma cell, an haemangioendothelioblastoma cell, an haemangioendothelioma cell, an haemangiopericytoma cell, an haematolymphangioma cell, an haemocytoblastoma cell, an liaemocytoma cell, a hairy cell leukaemia cell, a hamartoma cell, an hepatocarcinoma cell, an hepatocellular carcinoma cell, an hepatoma cell, an histoma cell , a Hodgkin's disease cell, an hypernephroma cell, an infiltrating cancer cell, an infiltrating ductal cell carcinoma cell, an insulinoma cell, a juvenile angiofibroma cell, a Kaposi sarcoma cell, a kidney tumour cell, a large cell lymphoma cell, a leukemia cell, a chronic leukemia cell, an acute leukemia cell, a lipoma cell, a liver cancer cell, a liver metastases cell, a Lucke carcinoma cell, a lymphadenoma cell, a lymphangioma cell, a lymphocytic leukaemia cell, a lymphocytic lymphoma cell, a lymphocytoma cell, a lymphoedema cell, a lymphoma cell, a lung cancer cell, a malignant mesothelioma cell, a malignant teratoma cell, a mastocytoma cell, a medulloblastoma cell, a melanoma cell, a meningioma cell, a mesothelioma cell, a metastatic cell, a metastasis cell, a metastatic spread cell, a Morton's neuroma cell, a multiple myeloma cell, a myeloblastoma cell, a myeloid leukemia cell, a myelolipoma cell, a myeloma cell, a myoblastoma cell, a myxoma cell, a nasopharyngeal carcinoma cell, a neoplastic cell, a nephroblastoma cell, a neuroblastoma cell, a neurofibroma cell, a neurofibromatosis cell, a neuroglioma cell, a neuroma cell, a non-Hodgkin's lymphoma cell, an oligodendroglioma cell, an optic glioma cell, an osteochondroma cell, an osteogenic sarcoma cell, an osteosarcoma cell, an ovarian cancer cell, a Paget's disease of the nipple cell, a pancoast tumour cell, a pancreatic cancer cell, a phaeochromocytoma cell, a pheochromocytoma cell, a plasmacytoma cell, a primary brain tumour cell, a progonoma cell, a prolactinoma cell, a renal cell carcinoma cell, a retinoblastoma cell, a rhabdomyosarcoma cell, a rhabdosarcoma cell, a solid tumor cell, sarcoma cell, a secondary tumour cell, a seminoma cell, a skin cancer cell, a small cell carcinoma cell, a squamous cell carcinoma cell, a strawberry haemangioma cell, a T-cell lymphoma cell, a teratoma cell, a testicular cancer cell, a thymoma cell, a trophoblastic tumour cell, a tumourigenic cell, a tumour initiation cell, a tumour progression cell, a vestibular schwannoma cell, a Wilm's tumour cell, or a combination thereof.

[00073] As defined herein, "contacting" means that a compound of the present invention is introduced into a sample containing the cell in a test tube, flask, tissue culture, chip, array, plate, microplate, capillary, or the like, and incubated at a temperature and time sufficient to permit inhibition of cell growth, inhibition of cell proliferation and/or induction of cell death. Methods for contacting the samples with the compounds are known to those skilled in the art and may be selected depending on the type of assay protocol to be run. Incubation methods are also standard and are lαiown to those skilled in the art.

[00074] In another embodiment, the term "contacting" means that a compound of the present invention is introduced into a subject receiving treatment, and the compound is allowed to come in contact with the cancer cell in vivo.

[00075] As used herein, the term "treating" includes preventative as well as disorder remitative treatment. As used herein, the term "inhibiting" has its commonly understood meaning of lessening or decreasing. As used herein, the term "progression" means increasing in scope or severity, advancing, growing or becoming worse. As used herein, the term "delaying" means postponing, setting back, slowing down.

[00076] As used herein, the term "administering" refers to bringing a subject in contact with a compound of the present invention. As used herein, administration can be accomplished in vitro, i.e. in a test tube, or in vivo, i.e. in cells or tissues of living organisms, for example humans. In one embodiment, the present invention encompasses administering the compounds of the present invention to a subject.

[00077] In one embodiment, the compounds of the present invention are selective inhibitors of cancer cell growth and proliferation, i.e. they inhibit the growth of cancer cells while having little effect on normal cells. The term "normal cell" is defined herein as a biological cell that does not express a malignant phenotype. A normal phenotype is defined herein as a phenotype which is not malignant, i.e. not characterized by an aberrant structure of the nucleus, nucleolus and cytoplasm, nucleus-to-plasma ratio, nuclear and chromosomal aberrations decreased cytoplasmic-nuclear ratio, an irregular chromatin network, larger nucleoli than normal, etc.

As defined herein, the term "selective" or "selectively" means that the compounds of the present invention are effective against cancer cells, while having no effect or a minimal effect on normal cells. As defined herein the term "no effect" also includes a small or minimal effect, for example a 1-20% inhibition of growth of normal cells. In one embodiment, the compounds of the present invention have no effect on the growth of normal cells. In another embodiment, the compounds of the present invention inhibit the growth of normal cells by 1-5%. In another embodiment, the compounds of the present invention inhibit the growth of normal cells by 5-10%. In another embodiment, the compounds of the present invention inhibit the growth of normal cells by 10-20%. Methods for testing cell growth inhibition, inhibition of cell proliferation and/or induction of cell death are known to those skilled in the . Any apropriate assay protocol known in the art may be used. In a non-limiting embodiment of the present invention, cell growth inhibition may be tested by a Methylene Blue assay, in accordance with embodiments of the present invention. In a further non-limiting embodiment of the present invention, cell growth inhibition may be tested by a Sulforhodamine B assay, in accordance with embodiments of the present invention.

[00078] The term " Effective amounts" are those amounts of a candidate substance effective to reproducibly decrease, reduce, inhibit or otherwise abrogate the growth of a cancer cell in comparison to levels in untreated cells.

[00079] In one embodiment, the methods of the present invention comprise administering a compound according to any of formulas I-XV as the sole active ingredient. However, also encompassed within the scope of the present invention are methods for inhibition of cancer cell growth, for inhibition of cell proliferation, for induction of cell death, for treatment of cancer and for delaying the progression of cancer, which comprise administering the compound in combination with one or more therapeutic agents. These agents may include any anticancer drug, cytotoxic drug, differentiation agents or any other agent which is useful in inhibition of cancer cell growth, inhibition of cell proliferation, induction of cell death, treatment of cancer and/or delaying the progression of cancer.

[00080] In another embodiment, the present invention further relates to the use of a pharmaceutical composition comprising a) as an active ingredient one or more compounds represented by the structure of any of formulas I-XV; and b) a pharmaceutically acceptable carrier in the treatment of cancer.

[00081] As used herein, "pharmaceutical composition" means therapeutically effective amounts of the compounds of the present invention, together with suitable diluents, preservatives, solubilizers, emulsifiers, adjuvant and/or carriers. A "therapeutically effective amount" as used herein refers to that amount which provides a therapeutic effect for a given condition and administration regimen. Such compositions are liquids or Lyophilized or otherwise dried formulations and include diluents of various buffer content (e.g., Tris-HCL, acetate, phosphate), pH and ionic strength, additives such as albumin or gelatin to prevent absorption to surfaces, detergents (e.g., Tween 20, Tween 80, Pluronic F68, bile acid salts), solubilizing agents (e.g., glycerol, polyethylene glycerol), anti-oxidants (e.g., ascorbic acid, sodium metabisulfite), preservatives (e.g., Thimerosal, benzyl alcohol, parabens), bulking substances or tonicity modifiers (e.g., lactose, mannitol), covalent attachment of polymers such as polyethylene glycol to the protein, complexation with metal ions, or incorporation of the material into or onto particulate preparations of polymeric compounds such as polylactic acid, polglycolic acid, hydrogels, etc, or onto liposomes, microemulsions, micelles, unilamellar or multilamellar vesicles, erythrocyte ghosts, or spheroplasts. Such compositions will influence the physical state, solubility, stability, rate of in vivo release, and rate of in vivo clearance. Controlled or sustained release compositions include formulation in lipophilic depots (e.g., fatty acids, waxes, oils).

[00082] Also comprehended by the invention are particulate compositions coated with polymers (e.g., poloxamers or poloxamines). Other embodiments of the compositions of the invention incorporate particulate forms, protective coatings, protease inhibitors or permeation enhancers for various routes of administration, including parenteral, pulmonary, nasal and oral. In one embodiment of the present invention, the pharmaceutical composition is administered parenterally, paracancerally, transmucosally, transdermally, intramuscularly, intravenously, intradermally, subcutaneously, intraperitonealy, intraventricularly, intracranially and intratumorally.

[00083] Further, as used herein "pharmaceutically acceptable carriers" are well known to those skilled in the art and include, but are not limited to, 0.01-0.1M and preferably 0.05M phosphate buffer or 0.8%> saline. Additionally, such pharmaceutically acceptable carriers may be aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media.

[00084] Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers such as those based on Ringer's dextrose, and the like. Preservatives and other additives may also be present, such as, for example, antimicrobials, antioxidants, collating agents, inert gases and the like.

[00085] Controlled or sustained release compositions include formulation in lipophilic depots (e.g. fatty acids, waxes, oils). Also comprehended by the invention are particulate compositions coated with polymers (e.g. poloxamers or poloxamines) and the compound coupled to antibodies directed against tissue-specific receptors, ligands or antigens or coupled to ligands of tissue-specific receptors.

[00086] Other embodiments of the compositions of the invention incorporate particulate forms, protective coatings, protease inhibitors or permeation enhancers for various routes of administration, including parenteral, pulmonary, nasal and oral.

[00087] Compounds modified by the covalent attachment of water-soluble polymers such as polyethylene glycol, copolymers of polyethylene glycol and polypropylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone or polyproline are known to exhibit substantially longer half- lives in blood following intravenous injection than do the corresponding unmodified compounds Such modifications may also increase the compound's solubility in aqueous solution, eliminate aggregation, enhance the physical and chemical stability of the compound, and greatly reduce the immunogenicity and reactivity of the compound. As a result, the desired in vivo biological activity may be achieved by the administration of such polymer-compound abducts less frequently or in lower doses than with the unmodified compound.

[00088] In yet another embodiment, the pharmaceutical composition can be delivered in a controlled release system. For example, the agent may be administered using intravenous infusion, an implantable osmotic pump, a transdermal patch, liposomes, or other modes of administration. In one embodiment of the present invention, a pump may be used. In another embodiment, polymeric materials can be used. In yet another embodiment, a controlled release system can be placed in proximity to the therapeutic target, i.e., the brain, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984)). Preferably, a controlled release device is introduced into a subject in proximity to the site of inappropriate immune activation or a tumor. Other controlled release systems are discussed in the review by Langer (Science 249:1527-1533 (1990)).

[00089] The pharmaceutical preparation can comprise a compound of any of formulas I-XV alone, or can further include a pharmaceutically acceptable carrier, and can be in solid or liquid form such as tablets, powders, capsules, pellets, solutions, suspensions, elixirs, emulsions, gels, creams, or suppositories, including rectal and urethral suppositories. Pharmaceutically acceptable carriers include gums, starches, sugars, cellulosic materials, and mixtures thereof. The pharmaceutical preparation containing the active compound can be administered to a subject by, for example, subcutaneous implantation of a pellet; in a further embodiment, the pellet provides for controlled release of the active compound over a period of time. The preparation can also be administered by intravenous, intraarterial, or intramuscular injection of a liquid preparation, oral administration of a liquid or solid preparation, or by topical application. Administration can also be accomplished by use of a rectal suppository or a urethral suppository.

[00090] The pharmaceutical preparations of the invention can be prepared by known dissolving, mixing, granulating, or tablet-forming processes. For oral administration, the active compound or their physiologically tolerated derivatives such as salts, esters, N-oxides, and the like are mixed with additives customary for this purpose, such as vehicles, stabilizers, or inert diluents, and converted by customary methods into a suitable form for administration, such as tablets, coated tablets, hard or soft gelatin capsules, aqueous, alcoholic or oily solutions. Examples of suitable inert vehicles are conventional tablet bases such as lactose, sucrose, or cornstarch in combination with binders like acacia, cornstarch, gelatin, or with disintegrating agents such as cornstarch, potato starch, alginic acid, or with a lubricant like stearic acid or magnesium stearate.

[00091] Examples of suitable oily vehicles or solvents are vegetable or animal oils such as sunflower oil or fish-liver oil. Preparations can be effected both as dry and as wet granules. For parenteral administration (subcutaneous, intravenous, intraarterial, or intramuscular injection), the compounds of the present invention or their physiologically tolerated derivatives such as salts, hydrates and the like are converted into a solution, suspension, or emulsion, if desired with the substances customary and suitable for this purpose, for example, solubihzers or other auxiliaries. Examples are: sterile liquids such as water and oils, with or without the addition of a surfactant and other pharmaceutically acceptable adjuvants. Illustrative oils are those of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, or mineral oil. In general, water, saline, aqueous dextrose and related sugar solutions, and glycols such as propylene glycols or polyethylene glycol are preferred liquid carriers, particularly for injectable solutions.

[00092] The preparation of pharmaceutical compositions which contain an active component is well understood in the art. In one embodiment of the present invention, such compositions are prepared as aerosols delivered to the nasopharynx or as injectables, either as liquid solutions or suspensions, however, solid forms suitable for solution in, or suspension in, liquid prior to injection can also be prepared. The preparation can also be emulsified. The active therapeutic ingredient is often mixed with excipients that are pharmaceutically acceptable and compatible with the active ingredient. Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol, or the like and combinations thereof.

[00093] In addition, if desired, the composition can contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents, which enhance the effectiveness of the active ingredient.

[00094] An active component can be formulated into the composition as neutralized pharmaceutically acceptable salt forms. Pharmaceutically acceptable salts include the acid addition salts (for e.g. with amine groups) which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed from the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, 2-ethylamino ethanol, histidine, procaine, and the like.

[00095] For topical administration to body surfaces using, for example, creams, gels, drops, and the like, the compounds of the present invention or their physiologically tolerated derivatives such as salts, hydrates, and the like are prepared and applied as solutions, suspensions, or emulsions in a physiologically acceptable diluent with or without a pharmaceutical carrier.

[00096] In another embodiment, the active compound can be delivered in a vesicle, in particular a liposome (see Langer, Science 249:1527-1533 (1990); Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez- Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; see generally ibid).

[00097] For use in medicine, the salts of the compounds will be pharmaceutically acceptable salts. Pharmaceutically acceptable salts include the acid addition salts which are formed by the reaction of free amino groups with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Other salts may, however, be useful in the preparation of the compounds according to the invention or of their pharmaceutically acceptable salts. Suitable pharmaceutically acceptable salts of the compounds of this invention include acid addition salts, which may, for example, be formed by mixing a solution of the compound according to the invention with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulphuric acid, methanesulphonic acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, oxalic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. Salts formed from free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, 2-ethylamino ethanol, histidine, procaine, and the like. [00098] The following examples are presented in order to more fully illustrate certain embodiments of the invention. They should in no way, however, be construed as limiting the broad scope of the invention.

EXPERIMENTAL DETAILS SECTION

Materials and methods.

Materials.

[0099] Cell lines were from American Type Culture Collection, Manassas, VA, USA. Growth media, fetal calf serum, donor horse serum, mycoplasma test kits, bovine insulin and PBS (Dulbecco's phosphate buffered saline) were from Biological Industries, Kibbutz Beit Haemek, Israel. Fetal bovine serum was from Gibco, Grand Island, NY, USA. Sulforhodamine B, sodium butyrate, hemoglobin, Genistein, 3,3',5,5'-tetramethylbenzidine, phorbol 12-myristate 13-acetate, May-Grunwald- Giemsa stain, Nile Red, DAPI (4'6'-diamidino-2-phenylindole dihydrochloride), microtubules from calf brain and monoclonal primary antibody to α-tubulin (clone DM 1A) were from Sigma-Aldrich, St. Louis, MO, USA. Mitomycin-C was from Kyowa Hakko Kogyo Co., Tokyo, Japan. Highly purified tubulin was from Cytoskeleton, Denver, CO, USA. Cy3 -conjugated (cyanine) goat anti-mouse secondary antibodies were from Jackson ImmunoResearch Laboratories, West Grove, PA, USA. Alexa Fluor 488 conjugated goat anti-mouse secondary antibodies were from Molecular Probes, Eugene, OR, USA. Nitroblue tetrazolium chloride was from Merck KGaA, Darmstadt, Germany. Gleevec™ Imatinib mesylate was from Novartis International AG, Basel, Switzerland. Hydromount was from National Diagnostic, Atlanta, Georgia, USA. ZTQ 1001 -ZTQ 1008, as summarized in Figure 1, were from a commercially available chemical library (ChemBridge Corporation; San Diego, CA, USA). Other chemicals, reagents and supplies were obtained from standard sources. All chemicals including water were tissue culture grade whenever needed; otherwise chemicals were at least reagent grade.

Cell culture standard growth conditions.

[00100] All cell lines were grown in 5% CO₂ at 37°C in a fully humidified atmosphere and were routinely tested for mycoplasma contamination using an EZ-PCR mycoplasma test kit. All growth media were supplemented with 100 U/ml penicillin G, 0.1 mg/ml streptomycin sulfate and 0.25 μg/ml amphotericin B.

[00101] CaCO2 cells (human colorectal adenocarcinoma) were grown in MEM Eagle medium supplemented with 20% fetal calf serum, 2 mM L-glutamine and 1 mM sodium pyruvate. CCD- 33 Co cells (normal primary colonocytes) were grown in MEM Eagle medium supplemented with 10%) fetal calf serum, 2 mM L-glutamine and 1 mM sodium pyruvate. HL-60 cells (acute promyelocytic leukemia) were grown in Iscove's modified Dulbecco's medium supplemented with 20% fetal calf serum and 4 mM L-glutamine. HT-29 (human colorectal adenocarcinoma) cells were grown in McCoy's medium supplemented with 10% fetal calf serum and 1.5 mM L-glutamine while HT-29* cells (a higher passage-number sub-clone with shorter doubling time) were grown in DMEM medium supplemented with 10% fetal calf serum and 2 mM L-glutamine. K-562 cells (human erythroleukemia) were grown in RPMI 1640 medium supplemented with 10%> fetal calf serum and 2 mM L-glutamine. LL/2 cells (mouse Lewis lung carcinoma) were grown in DMEM medium supplemented with 10% fetal bovine serum, 4 mM L-glutamine and 1.5 g/L sodium bicarbonate. MCF-7 cells (human breast cancer) were grown in MEM Eagle medium supplemented with 10% fetal calf serum, 2 mM L-glutamine, 1 mM sodium pyruvate and 0.25 U/ml bovine insulin (the MCF-7 cells used in all experiments were a higher passage number sub-clone). MIA-PaCa2 cells (pancreatic cancer) were grown in DMEM medium supplemented with 10% fetal calf serum, 2.5% donor horse serum and 4 mM L-glutamine. Rat-2 cells (rat fibroblast) were grown in DMEM medium supplemented with 10% fetal calf serum and 2 mM L-glutamine. SW-480 cells (human colorectal adenocarcinoma) were grown in DMEM medium supplemented with 10% fetal calf serum and 2 mM L-glutamine. U-937 cells (histiocytic lymphoma) were grown in RPMI 1640 supplemented with 10% fetal bovine serum, 10 mM HEPES, 1 mM sodium pyruvate, 2 mM L-glutamine and 4.5 g/L glucose. WiDr cells (human colorectal adenocarcinoma) were grown in MEM Eagle medium supplemented with 10% fetal calf serum, 2 mM L-glutamine and 1 mM sodium pyruvate.

Growth inhibition I (Methylene Blue assay).

[00102] Cells were seeded on day 1 into 384 well microtiter plates at concentration of 6,000 cells per well for the HT-29* cell line, and 10,000 cells per well for the WiDr and CCD-33Co cell lines. The following day, a plate for each cell line was fixed and kept at 4°C. Those plates were used to represent the cell density at time 0 of treatment. Cells were then treated with ZTQ1001-ZTQ1005, respectively, at various concentrations. The cells were exposed to treatment for 72 h with partially refreshment of media every 24 h. At the end of the treatment, cells were fixed and stained using the Methylene Blue assay (H. Ben-Bassat et al. 1997. Cancer Research, 57, 3741-3750.) as follows: Cells were fixed with 0.05% glutaraldehyde for 10 minutes at room temperature and then washed with water. The cells were then stained for 1 h with 0.1% methylene blue in 0.1M borate buffer pH=8.5. Excess dye was washed extensively with water and the plates were left to dry. The dye was eluted in 0.1 N HCl, 30 μl per well, for 1 h at 37°C, and the optical density was measured at 630 nm.

Growth inhibition H (Sulforhodamine B assay). [00103] HT-29* cells and CCD-33Co cells were seeded into 384 well microtiter plates with 6,500 HT-29* cells and 3,000 CCD-33Co cells per well respectively. The next day, 2-fold dilution series of each drug compound, ZTQ 1001 -ZTQ 1008, were individually introduced to the cells, and a sample plate was fixed to calculate the cell density at "time zero" of the experiment. The HT-29* cells were incubated with compounds for 72 h while the CCD-33Co cells were incubated with compounds for 216 h with media refreshing after 72 h. After incubation, the media was aspirated and the cells were fixed with cold 10%) (w/v) trichloroacetic acid (final concentration). The plates were then incubated for 60 minutes at 4°C after which the fixative was removed. The plates were washed 5 times with tap water and air-dried over night. The following day, 30 μl of 0.4%) (w/v) Sulforhodamine B in 1 % acetic acid was added to each well and the plates were incubated for 10 minutes at room temperature. After staining, unbound dye was removed by washing 3 times with 1% acetic acid and the plates were air-dried. Subsequently, bound stain was solubilized with 30 μl 10 mM trizma base and the optical density was read at 490 nm with reduction of background at 630 nm.

Cell line Panel.

[00104] Each of the cell lines CaCO2, CCD-33Co, HT-29*, MCF-7, MIA-PaCa2, Rat-2, SW-480 and WiDr cells were grown under standard conditions, seeded into microtiter plates, treated with increasing concentrations of ZTQ 1001 -ZTQ 1005, respectively, and assayed for total cell protein after drug treatment, all as described under "Growth inhibition II". In a parallel experiment, the leukemic cell lines K-562, HL-60 and U-937 were seeded into 96 well microtiter plates with 20,000 K-562 and U-937 cells and 40,000 HL-60 cells per well respectively. 2.5-fold dilution series (from 0.01 to 1 μM) of each drug compound, ZTQ1001-ZTQ1005, were individually introduced to the cells. The cells were then treated and stained as described under "Growth inhibition II" with adjustments of reagent volumes according to the larger well format.

Soft Agar. [00105] 1,000,000 HT-29* cells were seeded into 5-6 ml growth media and incubated under standard growth conditions for 24 h. The cells were then treated for 72 h with each investigated compound, ZTQ1001-1008, at indicated concentrations and under standard growth conditions. The cells were then trypsinized, counted, and re-suspended to a concentration of 10,000 cells per 50μl. Cell viability was verified with Trypan Blue and cell concentration was calculated for the viable cells only. 10,000 cells were then suspended in 1.5 ml 0.5% soft agar in complete medium and plated onto a 1.0 % agarose underlayer in 35x10 mM Petri plates. The plates were incubated at standard growth conditions for 10-13 days (as indicated) with addition of 200-300 μl of media to the top of each plate every 3-4 days. The colonies were then counted and compared to an untreated control.

Terminal differentiation of K-562 cells (erythroid cell differentiation).

[00106] Exponentially growing K-562 leukemic cells at an initial concentration of 150,000 cells/ml were incubated with ZTQIOOI, Gleevec™, or without compound addition for 3 days at the indicated concentrations without medium changes. 100,000 cells from each treatment were removed, pelleted by centrifugation, and washed with

PBS. The cells were then re-suspended in 100 μl of water, vortexed, freeze/thawed 3 times, and thereby lysed. The lysate was stored at -80°C until usage. The lysates were thawed and vortexed, and cellular debris was removed by centrifugation. 50 μl of lysates was reacted with 200 μl of 5 mg/ml 3,3',5,5'-tetramethylbenzidine, 0.5%o hydrogen peroxide in 50% acetic acid. The assay mixture was incubated for 20 min in the dark after which the optical density at 515 nm was measured. The assay was carried out in a

96 well microtiter plate and a 0 to 20- μg dilution series of purified hemoglobin were used for the standard curve.

Nitrobluetetrazolium test.

[00107] U-937 leukemic cells at an initial concentration of lxlO⁵ cells/ml were incubated under standard growth conditions with 1 μM all-trans retinoic acid and 0.1-0.5 μM ZTQIOOI for 6 days. Then lxlO⁶ cells in 1 ml of growth media were incubated at 37°C for 30 min in the presence of 0.1% nitroblue tetrazolium chloride and 100 ng of phorbol 12-myristate 13-acetate. After incubation, the cells were cytospinned and the slides stained with May-Grunwald-Giemsa stain. Cells were scored for the presence of blue-black formazan granules, while treatment with 0.8 mM sodium butyrate was used for positive control.

Nile Red test.

[00108] 3xl0⁵ MCF-7 cells were plated on sterile coverslips in 100-mm² dishes and grown for 96 h under standard growth conditions in the presence of either 2.5 mM sodium butyrate (in water) or 2 μM ZTQIOOI (in 0.06% DMSO). The cells were then simultaneously fixed and permeabilized in PBS containing 3% paraformaldehyde and 0.5%) Triton X-100 for 2 min, post-fixed in 3% paraformaldehyde for 20 min, and incubated for 5 min at room temperature with the lipid stain, Nile Red (1:1000 dilution of a 1 mg/ml acetone solution). Coverslips were rinsed in PBS and mounted with Hydromount. Images were obtained using an Olympus BX51 microscope (x60 objective).

Micronucleation.

[00109] MCF-7 cells were incubated for 72 h under standard growth conditions with lμM ZTQIOOI or 2.5 mM sodium butyrate and then fixed with ice-cold methanol. For DNA staining, cells were incubated with PBS containing 4'6'-diamidino-2-phenylindole dihydro chloride at a dilution of 1 :20,000 for 30 min and rinsed with two changes of PBS.

Microtubule disruption.

[00110] In one experiment, MCF-7 cells were incubated for 30 min in growth media supplemented with 2 μM ZTQIOOI. In another experiment, MCF-7 cells were incubated for 72 h in growth media supplemented with 1 μM ZTQIOOI or 2.5 mM sodium butyrate. In another experiment, MCF-7 cells, which overexpressed FTaseβ (farnesyltransferase subunit β), were incubated in growth media supplemented with 2 μM ZTQ 1005 for 30 min. All experiments were carried out under standard growth conditions and control experiments were carried out in growth media without drug supplementation. For tubulin labeling, the cells were fixed with cold 100%) methanol at -20°C for 7 min. The fixed cells were rinsed with PBS, incubated for 30 min with primary antibody to α-tubulin (1 :500) in PBS, rinsed three times in PBS, and then incubated with Cy3 -conjugated or Alexa Fluor 488 conjugated goat anti-mouse secondary antibodies for 30 min at room temperature. The labeled coverslips were rinsed in PBS, mounted with Hydromount and examined using an Olympus BX51 microscope (x60 objective).

Inhibition of microtubule polymerization.

[00111] Microtubules from calf brain were depolymerized according to the manufacturer's protocol. Tubulin heterodimers (10 μM) were incubated with 2 μM ZTQIOOI, 1 μM Taxol or 2 μM Vincristine, respectively, in PEMT buffer (100 mM PIPES, pH 7.5, 1 mM EGTA, 1 mM MgCl2 and 0.05% Triton-X-100) containing 1 mM GTP in a total volume of 100 ml at 37°C for 1 h. Samples (75 ml) were then transferred to centrifugal 0.22-μm pore size filter units, which had been previously washed with 200 ml PEM buffer (100 mM PIPES, pH 7.5, 1 mM EGTA and 1 mM MgCl₂). Recovered microtubules on the filters were stained with 50 ml of amido black solution (0.1%) naphthol blue black, 45% methanol and 10% acetic acid) for 2 min. Unbound dye was removed by two additions of 200 ml of destaining solution (90% methanol and 2% acetic acid). The microtubule-bound dye was then eluted by incubation with elution solution (25 mM NaOH, 0.05 mM EDTA and 50% ethanol) for 10 min. The elution solution was then transferred to new Eppendorf tubes and the absorbance measured at 600 nm. In a parallel assay, highly purified tubulin (1 mg/ml) was incubated with 0.5 μM Taxol in 80 mM PIPES, pH6.9, 1 mM EGTA and 1 mM MgCl₂ and 1 mM GTP for 1 hour in order to induce polymerization and stabilization of the microtubules. The Taxol stabilized microtubules were then treated with increasing concentrations of ZTQIOOI and ZTQ 1005 as indicated. Detection of microtubules was performed as described above.

Ex- Vivo colon tumor growth inhibition.

[00112] Two ex- vivo experiments were conducted using the HT-29 colon tumor model. In brief, HT-29* cells were incubated for three days in growth media containing the experimental compounds as described below. The growth media containing the experimental compounds were changed daily. On the fourth day the cells were harvested and suspended in PBS to a final concentration of 2.5xl0⁷ cells per ml. Immediately following harvest and suspension 0.2 ml of HT-29* cells from each test group were injected subcutaneously into the dorsal side of 7-10 ICR CDl nude mice (5xl0⁶ cells per mouse). During the assay period tumor size and mice body weight were recorded twice a week. Tumor volume (mm³) was estimated according to the formula: length (mm) x [width (mm)]² x 0.5. In the first assay, HT-29* cells were incubated every day for three days with: 100 μM Genistein (positive control), 1 μM ZTQ1003, or medium alone. In the second assay, HT-29* cells were incubated every day for three days with: 100 μM Genistein (positive control), 2 μM ZTQ 1002, or medium alone.

Results:

Growth inhibition 1 (Methylene Blue assay).

[00113] ZTQ1001-ZTQ1005 showed growth inhibition of the HT-29* and WiDr cell lines (both human colorectal adenocarcinoma) and were not cytotoxic to the CCD-33Co cell line (normal colonocytes). The compounds showed dose dependency and were active below 2 μM. Percentage growth inhibition was calculated as: (Ti/C) x 100, where C = optical density in untreated cells (control growth), and Ti = optical density in test growth in the presence of each investigated compound. Graphs showing the effect on cell numbers are depicted in Figures 2A-2E.

Growth inhibition LI (Sulforhodamine B assay).

[00114] ZTQ 1001 -ZTQ 1008 showed growth inhibition of the HT-29* cell line (human colorectal adenocarcinoma) and were not cytotoxic to the CCD-33Co cell line (normal colonocytes, which in some experiments did not proliferate during the assay period). The compounds showed dose dependency and were active below 2 μM. Percentage growth inhibition was calculated as: [(Ti-Tz/(C-Tz)] x 100, where C = optical density in untreated cells (control growth), Tz = optical density at time zero, and Ti = optical density in test growth in the presence of each investigated compound. Graphs showing the effect on cell numbers are depicted in Figures 3 A-3D.

Cell line Panel. [00115] ZTQ 1001 -ZTQ 1005 showed growth inhibition of the CaCO2, HL-60, HT-29*, K-562, MCF-7, MIA-PaCa2, Rat-2, SW-480, U-937 and WiDr cell lines. Percentage growth inhibition for each experimental agent for each mentioned cell line were calculated as described under "Specific growth inhibition". GI50 values (the drug concentration resulting in a 50%o reduction in the net protein increase) for each experimental agent are shown in Figure 4.

Soft Agar.

[00116] ZTQ 1001 -ZTQ 1008 inhibited the colony formation of HT-29* cells (colonic adenocarcinoma) under non-solid support conditions (growth in soft agar). Figure 5 represents this inhibition of colony formation in soft agar by ZTQ 1001 -ZTQ 1008. ZTQ1001-ZTQ1008 thus inhibited the anchorage-independent growth ability of untreated HT-29* cells. Some malignant cells, such as HT-29, have lost their anchorage dependency and are able to form colonies when grown in agar. The inhibition of the ability of the HT-29* cells to grow in soft agar indicates the potential anti-cancer activity of ZTQ 1001 -ZTQ 1008.

Terminal differentiation of K-562 cells (erythroid cell differentiation).

[00117] The effect of 72 h treatment with 0.1 μM and 0.5 μM ZTQIOOI followed by 174 h retraction on hemoglobin production in K-562 cells is illustrated in Figure 6. ZTQIOOI induced differentiation in K-562 cells, which was detected by hemoglobin production. In control cells, a low basal level of hemoglobin was detected. 72 h treatment with ZTQIOOI or Gleevec™ resulted in comparable increases of hemoglobin levels. The stability and duration of the phenotype normalization is an important parameter of the drug activity. Differentiation induced after 72 h treatment with Gleevec™ was stable after treatment retraction for 138 h and decreased almost 2 fold after 174 h. Treatment with ZTQIOOI resulted in a completely different and unexpected mode of behavior. Hemoglobin production increased almost 3-fold after 90 h of treatment retraction and continued to increase up to almost 5-fold at 138 h of retraction. Only a minor decrease in hemoglobin level was detected at 174 h.

Nitrobluetetrazolium test.

[00118] Tetrazolium salts reduction to formazan by dehydrogenases and reductases is used as an indicator of mitochondrial metabolism and is a relevant test for differentiation in a number of cellular systems. The effect of ZTQIOOI on cell differentiation in U-937 human leukemia cells was examined by a nitroblue tetrazolium reduction test (Figure 7A-C). In non-treated control cells only few formazan deposits were detected (Figure 7A, a', a"). ZTQIOOI at a concentration of 300 nM induced massive increase in formazan deposition and as a result, the percentage of nitroblue tetrazolium positive cells increased (Figure 7B, b', b"). The number of such positive cells and intensity of the deposits were used as a criterion of cell differentiation. Sodium butyrate induces the phenotypic differentiation in many cellular systems (U-937, MCF-7, HT-29) and it was therefore used as a positive control for the test (Figure 7C, c', c"). The pattern of the formazan aggregates was different between ZTQIOOI and sodium butyrate treated cells.

ZTQIOOI induced formation of two types of the pattern: Dot-like (Figure 7B, b') and massive homogenous distribution (Figure 7B, b"). In the same experiment, sodium butyrate had a preference to trigger the formation of the second type of the pattern

(Figure 7C, c\ c").

Nile Red test.

[00119] Lipid droplets are found in a variety of differentiating systems and in the cytoplasm of normal mammary epithelium. Differentiation triggering in human breast cancer cell lines by a number of compounds is also associated with lipid drop accumulation in the cellular cytoplasm. Based on the previous technique, we used a fluorescent stain, Nile Red, to visualize the lipid drop formation and accumulation in MCF-7 cells in response to the treatment with ZTQIOOI (Figure 8B). Treatment with sodium butyrate was used as positive control (Figure 8C). Lipid droplet accumulation was weak or absent in the untreated control cells (Figure 8A). A dramatic increase in drops per cell as well as the percent of the droplet-positive cells was detected as a result of the ZTQIOOI and sodium butyrate treatments.

Micronucleation.

[00120] As a result of the treatment with ZTQIOOI, MCF-7 cells became much bigger than untreated MCF-7 cells, their spreading increased dramatically, and the shape of the nucleus changed. The specific pattern described in the literature as micronucleation was detected as a result of ZTQIOOI treatment (Figure 9 A). Sodium butyrate treatment did not lead to micronucleation of MCF-7 cells (Figure 9B) and we did not detect these nuclear changes in the control cultures either (Figure 9C). These results indicate that the potential mechanism of differentiation induced by ZTQIOOI and butyrate is different.

Microtubule disruption. [00121] The effects of ZTQIOOI and ZTQ 1005 on the microtubule network were studied on MCF-7 cells (Figure 10A-C and 11 A-B). Treatment of MCF-7 cells for 30 min as well as for 72 h with ZTQIOOI resulted in microtubule disruption (Figures 10A and 11 A). Microtubule disruption was likewise seen after 30 min treatment with ZTQ 1005 (data not shown). Sodium butyrate treatment did not result in such an effect (Figure 10B). The microtubule density and pattern after treatment with sodium butyrate was similar to the untreated control (Figure 10C).

Inhibition of microtubule polymerization.

[00122] ZTQlOOl's disruption of microtubules in living cells may be a result of a direct effect of the drug on the tubulin or of an indirect mode of activity. To determine whether

ZTQIOOI interferes directly or indirectly with microtubule stability, its effect on the microtubule formation and stability in vitro was examined. For this purpose ZTQIOOI

(2 μM) was added to a purified tubulin solution under polymerization-induction conditions. Controls used were Taxol (1 μM) for hyperstabilization, Vincristine (2 μM) for depolymerization, and an untreated sample. The effect of each treatment is illustrated in Figure 12, where the OD 600 nm represents the extent of microtubule polymerization.

ZTQIOOI addition resulted in a more than 3 -fold decrease in the degree of polymerization compared to the untreated control. Taxol addition resulted in an almost

3-fold increase in the microtubule polymerization compared to the untreated control, and Vincristine resulted in a 10-fold decrease. These results indicated that ZTQIOOI directly targets the microtubule stability.

[00123] ZTQIOOI and ZTQ 1005 depolymerized microtubules formed from highly purified tubulin that had been stabilized with Taxol. The effect of ZTQIOOI and ZTQ 1005 on Taxol-stabilized microtubules is illustrated in Figure 13.

Ex- ivo colon tumor growth inhibition.

[00124] Ex-vivo treatment of HT-29* cells with ZTQ1002 and-ZTQ1003 prior to the cells' subcutaneous implantation into nude mice slowed the growth of the resulting tumors when compared to implantation with untreated HT-29* cells. The tumor growth curves are illustrated in Figures 14 and 15. The tumor growth rate for the cells treated with ZTQ1002, ZTQ1003, or Genistein(^*) was significantly slower than for the untreated control (P<0.05, t-test). [00125] () Positive control: Bennink, M et al. 1998. In: Bailliere's Clinical Endocrinology and Metabolism, 12, 707-728. London: Bailliere Tindall press.

[00126] It will be appreciated by a person skilled in the art that the present invention is not limited by what has been particularly shown and described hereinabove. Rather, the scope of the invention is defined by the claims which follow:

Claims

WHAT IS CLAIMED IS:

1. A method of inhibiting the growth of a cancer cell, comprising the step of contacting said cell with a compound represented by the structure of formula I, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to inhibit the growth of said cancer cell

I wherein

/^R5 Ri is H, a C₁-C₄ linear or branched alkyl or — N , wherein

R-' R₅ and R5' are independently of each other H, a C1-C4 linear or branched alkyl, / K , or R₅ and R5' together with the nitrogen to which they are

< attached represent a .

Rβ is H, a C₁-C₄ linear or branched alkyl, hydroxy, alkoxy, halogen, carboxy or COORu, wherein Ru is a C₁-C₄ linear or branched alkyl;

R₇ and R₇' are independently of each other H, a C₁-C₄ linear or branched alkyl, hydroxy, alkoxy, halogen or carboxy;

R and R₃ are independently of each other H, a C1-C4 linear or branched alkyl,

^/ in

R₈ and R₈' are independently of each other H, O, a C1-C₄ linear or branched alkyl, __/ , -(CH₂) _nOR wherein n is an integer of 1-4 and R is H or a C₁-C₄ linear

or branched alkyl or Rs and R₈' together with the nitrogen to which they are attached represent a group of the formula

R is H, a C₁-C₄ linear or branched alkyl, or / K ;

Rio and Rio' are independently of each other H, a C₁-C₄ linear or branched alkyl, or — ^~~K ; and

R₄ is H, a C₁-C₄ linear or branched alkyl, phenyl or NO₂.

2. The method of claim 1, wherein Ri is CH .

The method of claim 1, wherein Ri

6. The method of claim 1, wherein R₂ is methyl.

The method of claim 1,

The method of claim 1, wherein R₂ is — N

9. The method of claim 1, wherein R₂

10. The method of claim 1, wherein >

11. The method of claim 1 , wherein R₃ is

12. The method of claim 1, wherein R₃ is

13. The method of claim 1, wherein R₃ is

14. The method of claim 1, wherein R₃ is — NR₈R₈' wherein one of R₈ and R₈' is H and the other is . ^^{^}^

^■x^

15. The method of claim 1, wherein R₃

16. The method of claim 1, wherein R₃

17. The method of claim 1, wherein R₃

18. The method of claim 1, wherein said compound is represented by the structure of formula II.

19. The method of claim 18, wherein R₇ and R - are both CH₃. 20. The method of claim 18, wherein one of Rio and RIO' is CH₃ and the other is 4-methylphenyl. 21. The method of claim 1, wherein said compound is represented by the structure of formula III.

22. The method of claim 1, wherein said compound is represented by the structure of formula IV.

23. The method of claim 22, wherein Re is H.

24. The method of claim 1, wherein said compound is represented by the structure of formula V.

25. The method of claim 1, wherein said compound is represented by the structure of formula NI.

26. The method of claim 25, wherein R_δ is COOCH₃.

27. The method of claim 25, wherein R₇ and R₇> are both CH₃.

28. The method of claim 1, wherein said compound is represented by the structure of formula NIL

vπ

29. The method of claim 1, wherein said compound is represented by the structure of formula NIII.

30. The method of claim 29, wherein R is H.

31. The method of claim 1, wherein said compound is represented by the structure of formula IX.

32. The method of claim 1, wherein said compound is represented by the structure of formula X.

33. The method of claim 32, wherein Rg is OCH₃. 34. The method of claim 32, wherein R₇ and R₇> are both CH₃.

35. The method of claim 1, wherein said compound is represented by the structure of formula XI.

XI

36. The method of claim 1, wherein said cancer cell is a colon cancer cell.

37. A method of inducing cell death, comprising the step of contacting a cell with a compound represented by the structure of formula I, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to induce the death of said cell

I wherein

/^R5 Ri is H, a C₁-C₄ linear or branched alkyl or — N , wherein

Rs' R₅ and R5' are independently of each other H, a C₁-C₄ linear or branched a, , _ ^R' , or R₃ and V together with the nitrogen to which they are

attached represent .

Rg is H, a C₁-C4 linear or branched alkyl, hydroxy, alkoxy, halogen, carboxy or

COOR₁₁ wherein Ru is a C₁-C₄ linear or branched alkyl;

R and R₇' are independently of each other H, a C₁-C₄ linear or branched alkyl, hydroxy, alkoxy, halogen or carboxy;

R₂ and R₃ are independently of each other H, a C₁-C₄ linear or branched alkyl,

Rs and Rs' are independently of each other H, O, a C1-C4 linear or branched -(CH₂) _nOR wherein n is an integer of 1-4 and R is H or a Cι-C linear

or branched alkyl, or R₈ and Rs' together with the nitrogen to which they are attached represent a group of the formula

R is H, a C1-C₄ linear or branched alkyl, or / K ;

Rio and Rio' are independently of each other H, a C1-C4 linear or branched alkyl, or — /^{^} ;

and R₄ is H, a C1-C₄ linear or branched alkyl, phenyl or NO .

38. The method of claim 37, wherein Ri is CH₃.

39. The method of claim 37, wherein Ri is NH₂.

40. The method of claim 37, wherein Ri

41. The method of claim 37, wherein Ri

42. The method of claim 37, wherein R₂ is met thnyyli

43. The method of claim 37, wherein R₂

44. The method of claim 37, wherein R₂ is — N

45. The method of claim 37, wherein R₂ is

46. The method of claim 37, wherein R₂ is

47. The method of claim 37, wherein R₃ is phenyl.

48. The method of claim 37, wherein R₃ is ⁹ wherein R₉ is ^ ■

49.

50. The method of claim 37, wherein R₃ is — NR₈Rs' wherein one of R₈ and R₈' is H and the other is . //

51. The method of claim 37, wherein R₃ is

52. The method of claim 37, wherein R₃ is

53. The method of claim 37, wherein R₃

54. The method of claim 37, wherein said compound is represented by the structure of formula II.

55. The method of claim 54, wherein R₇ and R_7' are both CH₃.

56. The method of claim 54, wherein one of Rio and RIO' is CH₃ and the other is 4-methylphenyl.

57. The method of claim 37, wherein said compound is represented by the structure of formula III.

58. The method of claim 37, wherein said compound is represented by the structure of formula IV.

59. The method of claim 58, wherein s is H.

60. The method of claim 37, wherein said compound is represented by the structure of formula V.

61. The method of claim 37, wherein said compound is represented by the structure of formula VI.

62. The method of claim 61, wherein 5 is COOCH₃. 63. The method of claim 61, wherein R₇ and Rr are both CH₃.

64. The method of claim 37, wherein said compound is represented by the structure of formula VII.

vπ

65. The method of claim 37, wherein said compound is represented by the structure of formula NIII.

66. The method of claim 65, wherein Rs is H.

67. The method of claim 37, wherein said compound is represented by the structure of formula IX.

68. The method of claim 37, wherein said compound is represented by the structure of formula X.

69. The method of claim 67, wherein s is OCH₃.

70. The method of claim 67, wherein R₇ and R₇> are both CH₃.

71. The method of claim 37, wherein said compound is represented by the structure of formula XI.

XI

72. The method of claim 37, wherein said cell is a cancer cell.

73. The method of claim 37, wherein said cell is a colon cancer cell.

74. A method of delaying the progression of cancer in a subject having cancer, comprising the step of administering to said subject a compound represented by the structure of formula I, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to delay the progression of cancer in said subject

wherein

X

Ri is H, a Cι-C linear or branched alkyl or — N . wherein

R₅'

Rs and R₅' are independently of each other H, a C₁-C₄ linear or branched alkyl, / K , or R₅ and R₅' together with the nitrogen to which they are

attached represent a group of the or — N o .

Rg is H, a C1-C4 linear or branched alkyl, hydroxy, alkoxy, halogen, carboxy or

COOR₁₁, wherein Ru is a C1-C4 linear or branched alkyl;

R₇ and R ' are independently of each other H, a C₁-C₄ linear or branched alkyl, hydroxy, alkoxy, halogen or carboxy; R and R₃ are independently of each other H, a C1-C4 linear or branched alkyl,

— N , wherein

Rs and R₈' are independently of each other H, O, a C₁-C₄ linear or branched alkyl,_/ K ,-(CH₂) „OR wherein n is an integer of 1-4 and R is H or a C₁-C₄ linear

R₉ is H, a C₁-C₄ linear or branched alkyl, or / K ;

Rio and Rio' are independently of each other H, a Cι-C₄ linear or branched alkyl, or /^"^;

and

R₄ is H, a C₁-C₄ linear or branched alkyl, phenyl or NO₂.

75. The method of claim 74, wherein Ri is CH₃. 76. The method of claim 74, wherein Ri is NH₂.

77. The method of claim 74, wherein Ri

78. The method of claim 74, wherein Ri

79. The method of claim 74, wherein R₂ is methyl.

80. The method of claim 74, wherein R₂ is

81. The method of claim 74, wherein R is

82. The method of claim 74, wherein R₂ is

83. The method of claim 74, wherein R₃ is phenyl.

84. The method of claim 74, wherein R₃ is ⁹ wherein R₉ is ------/ X .

85. The method of claim 74, wherein R₃ is — O — — C— OCH₃.

86. The method of claim 74, wherein R₃ is — NRsR₈' wherein one of R₈ and Rs' is H and the other is {^{~^}X ■

87. The method of claim 74, wherein R₃ is — — /~ °^_CH ₃ .

88. The method of claim 74, wherein R₃

89. The method of claim 74, wherein R₃

90. The method of claim 74, wherein said compound is represented by the structure of formula II.

91. The method of claim 90, wherein R₇ and R₇' are both CH₃.

92. The method of claim 90, wherein one of Rio and Rio' is CH₃ and the other is 4-methylphenyl.

93. The method of claim 74, wherein said compound is represented by the structure of formula III.

94. The method of claim 74, wherein said compound is represented by the structure of formula IV.

95. The method of claim 94, wherein R is H.

96. The method of claim 74, wherein said compound is represented by the structure of formula N.

97. The method of claim 74, wherein said compound is represented by the structure of formula VI.

98. The method of claim 97, wherein Rs is COOCH₃. 99. The method of claim 97, wherein R₇ and R₇- are both CH₃.

100. The method of claim 74, wherein said compound is represented by the structure of formula VII.

vπ 101. The method of claim 74, wherein said compound is represented by the structure of formula VIII.

102. The method of claim 101, wherein Re is H.

103. The method of claim 74, wherein said compound is represented by the structure of formula IX.

104. The method of claim 74, wherein said compound is represented by the structure of formula X.

105. The method of claim 104, wherein R is OCH₃.

106. The method of claim 104, wherein R₇ and R₇> are both CH₃.

107. The method of claim 74, wherein said compound is represented by the structure of formula XL

108. The method of claim 74, wherein said cancer is colon cancer.

109. A method of inhibiting the growth of a cancer cell, comprising the step of contacting said cell with a compound represented by the structure of formula XII, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to inhibit the growth of said cancer cell.

Ri

XII wherein

Ri is a C₁-C₄ linear or branched alkyl, or Ri

R₄ and R₅ are independently of each other H, a Cι-C linear or branched alkyl, a C₁-C₄ linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, CF₃ or NO₂;

R₆ is H or a C1-C₄ linear or branched alkyl;

R is H or a C1-C₄ linear or branched alkyl.

110. The method of claim 109, wherein Ri is CH₃.

111. The method of claim 109, wherein Ri is phenyl.

112. The method of claim 109, wherein R₂ is ethoxy and R₃ is H.

113. The method of claim 109, wherein R₂ is bromo and R₃ is H.

114. The method of claim 109, wherein R₂ is chloro and R₃ is H.

115. The method of claim 109, wherein K is methoxy and R5 is H.

116. The method of claim 109, wherein R4 is ethoxy and R₅ is H.

117. The method of claim 109, wherein said compound is represented by the structure of formula XIII:

XIII

118. The method of claim 109, wherein said compound is represented by the structure of formula XIV:

119. The method of claim 109, wherein said compound is represented by the structure of formula XV:

120. The method of claim 109, wherein said cancer cell is a colon cancer cell.

121. A method of inducing cell death, comprising the step of contacting a cell with a compound represented by the structure of formula XII, or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to induce the death of said cell.

Ri xn wherein

Ri is a C₁-C4 linear or branched alkyl, or Ri is

R₂ and R₃ are independently of each other H, a C₁-C₄ linear or branched alkyl, a C₁-C4 linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, COR, COOR, COOH, NHCOR, CF₃, NO₂, or R₂ and R₃ together with the benzene group to which they are attached represent a fused ring system represented by the structure:

R-₄ and R5 are independently of each other H, a C₁-C₄ linear or branched alkyl, a C1-C4 linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, CF₃, or NO ;

R_δ is H or a C1-C4 linear or branched alkyl;

R₇ and R₈ are independently of each other H, a C₁-C₄ linear or branched alkyl, halogen, hydroxy, alkoxy, thio, alkylthio, CF₃, or NO₂; and R is H or a C₁-C₄ linear or branched alkyl.

122 The method of claim 121, wherein Ri is CH₃. 123 The method of claim 121, wherein Ri is phenyl. 124 The method of claim 121, wherein R₂ is ethoxy and R₃ is H. 125 The method of claim 121, wherein R₂ is bromo and R₃ is H. 126 The method of claim 121, wherein R₂ is chloro and R₃ is H. 127 The method of claim 121, wherein R₄ is methoxy and R5 is H. 128 The method of claim 121, wherein R4 is ethoxy and R₅ is H. 129 The method of claim 121, wherein said compound is represented by the structure of formula XIII:

xm 130. The method of claim 121, wherein said compound is represented by the structure of formula XIN:

131. The method of claim 121, wherein said compound is represented by the structure of formula XN:

132. The method of claim 121, wherein said cell is a cancer cell. 133. The method of claim 121, wherein said cell is a colon cancer cell.

134. A method of delaying the progression of cancer in a subject having cancer, comprising the step of administering to said subject a compound represented by the structure of formula XII or its pharmaceutically acceptable salt, hydrate or any combination thereof, in an amount effective to delay the progression of cancer in said subject

XII wherein

Ri is a C₁-C₄ linear or branched alkyl, or Ri

R₂ and R₃ are independently of each other H, a C₁-C₄ linear or branched alkyl, a C1-C4 linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, COR, COOR, COOH, NHCOR, CF₃, NO₂, or R₂ and R₃ together with the benzene group to which they are attached represent a fused ring system represented by the structure:

R4 and R₅ are independently of each other H, a C₁-C₄ linear or branched alkyl, a C1-C4 linear or branched haloalkyl, halogen, hydroxy, alkoxy, phenoxy, thio, alkylthio, arylthio, CF₃ or NO₂;

Rs is H or a C₁-C₄ linear or branched alkyl; R₇ and R₈ are independently of each other H, a C₁-C₄ linear or branched alkyl, halogen, hydroxy, alkoxy, thio, alkylthio, CF₃, or NO₂; and R is H or a C1-C₄ linear or branched alkyl.

135 The method of claim 134, wherein Ri is CH₃.

136 The method of claim 134, wherein Ri is phenyl.

137 The method of claim 134, wherein R₂ is ethoxy and R₃ is H.

138 The method of claim 134, wherein R₂ is bromo and R₃ is H.

139 The method of claim 134, wherein R₂ is chloro and R₃ is H.

140 The method of claim 134, wherein R4 is methoxy and R₅ is H.

141 The method of claim 134, wherein R4 is ethoxy and R₅ is H.

142. The method of claim 134, wherein said compound is represented by the structure of formula XIII:

XIII

143. The method of claim 134, wherein said compound is represented by the structure of formula XIV:

144. The method of claim 134, wherein said compound is represented by the structure of formula XV:

145. The method of claim 134, wherein said cancer is colon cancer.