WO2011066606A1

WO2011066606A1 - Pyranoindole compounds and uses thereof

Info

Publication number: WO2011066606A1
Application number: PCT/AU2010/001609
Authority: WO
Inventors: Taj Khan; David Black; Naresh Kumar; George Jeoffreys
Original assignee: Novogen Research Pty Ltd
Priority date: 2009-12-01
Filing date: 2010-11-30
Publication date: 2011-06-09

Abstract

The present invention relates to pyranoindole compounds and compositions comprising same. The invention further relates to the use of pyranoindole compounds for the treatment of diseases associated with aberrant cell proliferation such as cancer.

Description

Pyranoindole compounds and uses thereof Technical Field

Background of the Invention

Despite ongoing intensive research and development into anti-cancer therapies cancer continues to take a heavy toll of lives. More than 10 million people around the world are diagnosed with cancer every year and it is estimated that this number could increase to as many as 20 million new cases a year by 2030. In addition to the devastating personal and social effects of cancer, the public health costs associated with cancer treatment are estimated to be growing at 10% to 20% per year.

The most common cancers worldwide include lung, breast, prostate, liver, colorectal and stomach cancers. A number of other cancers are increasing in prevalence, such as melanoma, ovarian cancer and cervical cancer. Chemotherapy is typically the preferred method of treatment for the majority of the above cancers, and may be used in conjunction with surgery and/or radiotherapy. However, in an increasing number of patients the cancers develop resistance to the chemotherapeutic agents indicated for the particular cancer meaning that in some patients the treatment options available are severely limited. Furthermore, a number of patients suffer from side effects associated with chemotherapeutic agents (for example neurotoxicity) which may further limit their use. Against this background there is an ever present need for new chemotherapeutic agents that will provide additional treatment options for patients suffering from cancer, and in particular those cancers which are the most prevalent.

The present inventors have surprisingly found that certain pyranoindole compounds possess antiproliferative activity, and hence are useful in the treatment of cancer.

Summary of the Invention

In a first aspect, the present invention provides a compound of the general formula (I):

wherein:

X is O, S or NR₉,

Y is O, S or NRs

Ri is selected from the group consisting of: hydrogen, hydroxy, halogen, optionally substituted aryl, optionally substituted heteroaryl, alkylaryl, C(0)OCi-C₆ alkyl, Ci-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halo Ci-C₆ alkyl, halo C₂-C₆ alkenyl, halo C₂-C₆ alkynyl, Ci-C₆ alkoxy and halo Ci-C₆ alkoxy,

R₂ is selected from the group consisting of: hydrogen, optionally substituted aryl, optionally substituted heteroaryl, Ci-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halo C|-C₆ alkyl, halo C₂-C₆ alkenyl, halo C₂-C₆ alkynyl, Ci-C₆ alkoxy and halo Ci-C_fr alkoxy,

R₃, R4, R5, R6 and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, Ci-C₆ alkoxy, amino, halogen, C_t-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halo Ci-C₆ alkyl, halo C₂-C₆ alkenyl, halo C₂-C₆ alkynyl and halo Ci-C₆ alkoxy,

Rg and R₉ are independently selected from the group consisting of: hydrogen, Q-Q alkyl and alkylaryl,

the drawing "™" represents either a single bond or a double bond, and salts and derivatives thereof.

X may be O.

Y may be NRg.

Ri may be selected from the group consisting of: hydrogen, halogen, optionally substituted aryl, optionally substituted heteroaryl, Ci-C₆ alkyl, halo Ci-C₆ alkyl, Ci-C₆ alkoxy and halo Ci-C₆ alkoxy.

Ri may be selected from the group consisting of: hydrogen, halogen, optionally substituted aryl, optionally substituted heteroaryl, CpC₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 13 halogens. In another embodiment, Ri may be selected from the group consisting of: hydrogen, halogen, optionally substituted aryl, Ci-C₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 13 halogens.

In yet another embodiment, R_\ may be selected from the group consisting of: hydrogen, halogen, phenyl, Ci-C₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens, and wherein the phenyl group may optionally be substituted with Ci -C₆ alkyl, d -C₆ alkoxy, hydroxy or halogen.

In a further embodiment, Ri may be selected from the group consisting of: halogen, phenyl, C1-C4 alkyl and halo Ci-C₄ alkyl, wherein the halo Ci -C₄ alkyl group contains between 1 and 4 halogens, and wherein the phenyl group may optionally be substituted with Ci-C₆ alkyl, Ci-C₆ alkoxy or hydroxy.

R₂ may be selected from the group consisting of: hydrogen, optionally substituted aryl, Ci-C₆ alkyl and halo Ci-C₆ alkyl wherein the halo Ci -C₆ alkyl group contains between 1 and 6 halogens.

In another embodiment, R₂ may be selected from the group consisting of: hydrogen, optionally substituted phenyl, Ci-C₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens.

In an alternative embodiment, R₂ may be selected from the group consisting of: hydrogen, optionally substituted phenyl and C| -C₆ alkyl.

R₃, R4, R5, ¾ and R₇ may be independently selected from the group consisting of: hydrogen, hydroxy, halogen, C] -C₆ alkyl and Ci-C₆ alkoxy.

In another embodiment, R3, R4, R5, Re and R₇ may be independently selected from the group consisting of: hydrogen, hydroxy, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy and halogen.

In a further embodiment, R₃, R4, R5, R5 and R₇ may be independently selected from the group consisting of: hydrogen, hydroxy, methyl, ethyl, methoxy, ethoxy and halogen.

R¾ and R may be independently selected from the group consisting of: hydrogen, methyl, ethyl, propyl and isopropyl.

In a second aspect, the present invention provides a pharmaceutical composition comprising a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined in the first aspect, and a pharmaceutically acceptable carrier, diluent and/or excipient. In a third aspect, the present invention provides a method for inhibiting the proliferation of cells, said method comprising contacting the cells with a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined in the first aspect.

In a fourth aspect, the present invention provides a method for the prevention and/or treatment of cancer in a subject in need thereof, said method comprising ^ administration to the subject of a therapeutically effective amount of a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined in the first aspect.

The cancer may be selected from the group consisting of: breast cancer, ovarian cancer, leukaemia, prostate' cancer, liver cancer, colorectal cancer, gastric cancer, pancreatic cancer, glioma, melanoma and lung cancer.

In another embodiment the cancer is selected from the group consisting of: breast cancer, ovarian cancer, leukaemia, non-androgen dependent prostate cancer, liver cancer, colorectal cancer, gastric cancer, pancreatic cancer, glioma, melanoma and lung cancer.

Irl another embodiment the cancer is selected from the group consisting of: ovarian cancer, leukaemia, non-androgen dependent prostate cancer, liver cancer, colorectal cancer, gastric cancer, pancreatic cancer, glioma, melanoma and lung cancer.

In a further embodiment the cancer is selected from the group consisting of:

leukaemia, non-androgen dependent prostate cancer, liver cancer, colorectal cancer, gastric cancer, pancreatic cancer, glioma, melanoma and lung cancer.

In still another embodiment the cancer is selected from the group consisting of:

colorectal cancer, breast cancer, gastric cancer, liver cancer and lung cancer.

In another embodiment the cancer is selected from the group consisting of:

colorectal cancer, gastric cancer, liver cancer and lung cancer.

. The cancer may be a non-hormone dependent cancer.

The cancer may be a non-androgen dependent cancer.

In a fifth aspect, the present invention provides the use of a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined in the first aspect in the manufacture of a medicament for inhibiting the proliferation of cells.

In a sixth aspect, the present invention provides a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined in the first aspect, for use in inhibiting the proliferation of cells.

In a seventh aspect, the present invention provides the use of a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined in the first aspect in the manufacture of a medicament for the prevention and/or treatment of cancer.

In an eighth aspect, the present invention provides a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined in the first aspect, for use in the prevention and/or treatment of cancer.

These and other aspects of the invention will become evident from the description and claims which follow.

Definitions

Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

In the context of this specification, the terms "treatment" and "treating" refer to any and all uses which remedy a condition, disease, disorder or symptoms thereof, or otherwise prevent, hinder or reverse the progression of a condition, disease, disorder or symptoms thereof, in any way whatsoever. Treatment may be for a defined period of time, or provided on an ongoing basis depending on the particular circumstances of any given individual.

In the context of this specification, the terms "prevent" and "prevention" refer to any and all uses which prevent the establishment or onset of a condition, disease, disorder or symptoms thereof in any way whatsoever.

In the context of this specification, the term "therapeutically effective amount" includes within its meaning a non-toxic amount of a compound of formula (I) sufficient to provide the desired therapeutic effect. The exact amount will vary from subject to subject depending on, for example, the age of the subject, their general health, the severity of the disorder being treated and the mode of administration. It is therefore not possible to specify an exact "therapeutically effective amount", however one skilled in the art would be capable of determining such an amount by routine trial and experimentation. In the context of this specification, the term "salts thereof is understood to include acid addition salts, anionic salts and zwitterionic salts, and in particular pharmaceutically acceptable salts.

In the context of this specification, the term "pharmaceutically acceptable" means that the compound to which it refers is suitable for use in contact with tissues of the body without undue toxicity, incompatibility, instability, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts include salts of acidic or basic groups present in compounds. Examples of salts include but are not limited to, those formed from: acetic, ascorbic, aspartic, benzoic, benzenesulfonic, citric, cinnamic, ethanesulfonic, fumaric, glutamic, glutaric, gluconic, hydrochloric, hydrobromic, lactic, maleic, malic, methanesulfonic, naphthoic, hydroxynaphthoic, naphthalenesulfonic, naphthalenedisulfonic, naphthaleneacrylic, oleic, oxalic, oxaloacetic, phosphoric, pyruvic, p-toluenesulfonic, tartaric, trifluoroacetic, triphenylacetic, tricarballylic, salicylic, sulphuric, sufamic, sulfanilic and succinic acid. Pharmaceutically acceptable salts also include alkali or alkaline earth metal salts of acidic groups (for example, sodium, potassium, calcium and magnesium salts) and also any other non-toxic metal salts.

In the context of this specification, the term "Ci-C₆ alkyl" is taken to include straight chain and branched chain monovalent saturated hydrocarbon radicals having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, secbutyl, tertiary butyl, pentyl, hexyl and the like.

In the context of this specification, the term "Ci-C alkyl" is taken to include straight chain and branched chain monovalent saturated hydrocarbon radicals having 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl and the like.

In the context of this specification, the term "C1-C6 alkoxy" is taken to include alkyl ether radicals of the formula OCi-C₆ alkyl, wherein Ci-C₆ alkyl is as defined above. Examples of Ci-C₆ alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, pentoxy and the like.

In the context of this specification, the term "C₂-C₆ alkenyl" is taken to include straight chain and branched chain monovalent hydrocarbon radicals having 2 to 6 carbon atoms and at least one carbon-carbon double bond, such as vinyl, propenyl, 2-methyl-2- propenyl, butenyl, pentenyl and the like. In the context of this specification, the term "C₂-C6 alkynyl" is taken to include straight chain and branched chain monovalent hydrocarbon radicals having 2 to 6 carbon atoms and at least one carbon-carbon triple bond, such as ethynyl, propynyl, butynyl, pentynyl, hexynyl and the like.

In the context of this specification, the term "halo Ci-C₆ alkyl" has the same meaning as "Ci-C₆ alkyl", except that one or more hydrogens of the Ci-C₆ alkyl group are replaced by a halogen, or combination of halogens. The term "halo Ci-C₆ alkyl" is to be considered in its broadest context and includes mono-halogenated alkyl groups (for example -C¾F, -CH2CH2CI and the like) through to perhalogenated alkyl groups (for example -CF₃, -CF₂CF₃ and the like). In an embodiment of the invention, the halo Ci-C₆ alkyl group is a perfluoroalkyl group,

In the context of this specification, the term "halo C₂-C₆ alkenyl" has the same meaning as "halo Ci-C₆ alkyl", except that the "halo C2-C6 alkenyl" group contains one or more carbon-carbon double bonds.

In the context of this specification, the term "halo C₂-C₆ alkynyl" has the same meaning as "halo Ci-C₆ alkyl", except that the "halo C2-C6 alkynyl" group contains one or more carbon-carbon triple bonds.

In the context of this specification, the term "halo Ci-C₆ alkoxy" has the same meaning as "Ci-C₆ alkoxy", except that one or more hydrogens of the Q-C6 alkoxy group are replaced by a halogen, or combination of halogens. The term "halo Ci-C₆ alkoxy" is to be considered in its broadest context and includes mono-halogenated alkoxy groups (for example -OCH₂F, -OCH2CH2CI and the like) through to perhalogenated alkoxy groups (for example -OCF₃, OCF2CF3 and the like).

In the context of this specification, the term "aryl" is taken to include monovalent aromatic radicals having between 6 and 30, or 6 and 25, or 6 and 20, or 6 and 15, or 6 arid 10 carbon atoms. Aryl groups include, but are not limited to phenyl, biphenyl, naphthyl, pyrenyl, anthracenyl and phenanthrenyl. The aryl group may be unsubstituted or optionally substituted by one or more of: Ci-C₆ alkyl, C2-C₆ alkenyl, C₂-C₆ alkynyl, halogen, hydroxy, Ci-C₆ alkoxy, silyloxy, amino, diCi-C₆ alkylamino and phenyl.

In the context of this specification, the term "alkylaryl" is understood to mean a substituent comprising an aryl group and an alkyl group, wherein the substituent is attached to the remainder of the molecule via the alkyl group. Examples of "alkylaryl" groups include benzyl, ethylphenyl and the like.

In the context of this specification, the term "heteroaryl" is taken to include monovalent aromatic radicals having between 5 and 20, or 5 and 15, or 5 and 10 atoms, wherein 1 to 6, or 1 to 5, or 1 to 4, or 1 to 3, or 1 or 2 atoms are heteroatoms selected from nitrogen, oxygen and sulfur. Examples of heteroaryl groups include, but are not limited to: furanyl, quinazolinyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, pyrazolyl, tetrazolyl, oxazolyl, isoxazolyl, isothiazolyl, thiazolyl, thienyl, imidazolyl, pyrazinyl, pyridazinyl, pyrimidinyl, pyridyl, triazolyl, benzothiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl and triazinyl. The heteroaryl group may be unsubstituted or optionally substituted by one or more of: Ci-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halogen, hydroxy, Ci-C₆ alkoxy, silyloxy, amino, diCi-C₆ alkylamino and phenyl.

In the context of this specification, the terms "halo" and "halogen" are taken to include fluoro, chloro, bromo and iodo.

In the context of this specification, the term "pharmaceutically acceptable derivative" refers to a derivative of the active compound that upon administration to the recipient is capable of providing directly or indirectly, the active compound. Thus, derivatives include solvates, esters, prodrugs and the like, and also compounds possessing leaving groups that can be cleaved in vivo to provide active compounds of the invention. Suitable leaving groups include but are not limited to: acyl, phosphate, sulfate and sulfonate.

Detailed Description of the Invention aspect, the present invention provides a compound of the general formula (I):

wherein:

X is O, S or NR_g,

Y is O, S or NR₈ Ri is selected from the group consisting of: hydrogen, hydroxy, halogen, optionally substituted aryl, optionally substituted heteroaryl, alkylaryl, C(0)OCi-C6 alkyl, Ci-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halo Ci-C₆ alkyl, halo C₂-C₆ alkenyl, halo C₂-C₆ alkynyl, Ci-C₆ alkoxy and halo Ci-C₆ alkoxyj

R₂ is selected from the group consisting of: hydrogen, optionally substituted aryl, optionally substituted heteroaryl, C]-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halo Ci-C₆ alkyl, halo C₂-C₆ alkenyl, halo C₂-C₆ alkynyl, Ci-C₆ alkoxy and halo Ci-C₆ alkoxy,

R₃, R4, RJ, 6 and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, Ci-C₆ alkoxy, amino, halogen, Ci-C₆ alkyl, C2-C₆ alkenyl, C₂-C6 alkynyl, halo Ci-C₆ alkyl, halo C₂-C₆ alkenyl, halo C₂-C₆ alkynyl and halo Ci-C₆ alkoxy,

R_g and R9 are independently selected from the group consisting of: hydrogen and Ci-C₆ alkyl and alkylaryl,

X may be O.

Y may be NR».

Ri may be selected from the group consisting of: hydrogen, halogen, optionally substituted aryl, optionally substituted heteroaryl, Ci-C₆ alkyl, halo Ci-C₆ alkyl, Ci-C₆ alkoxy and halo Ci-C₆ alkoxy. The halogen may be fluoro.

Ri may be selected from the group consisting of: hydrogen, halogen, optionally substituted aryl, optionally substituted heteroaryl, Ci-C₆ .alkyl and halo Ci-C₆ alkyl, wherein the halo C]-C6 alkyl group contains between 1 and 13 halogens. The halogen may be fluoro.

In another embodiment, Ri may be selected from the group consisting of: hydrogen, halogen, optionally substituted aryl, Ci-C₆ alkyl and halo Ci-C₆ alkyl, wherein the Ci-C₆ alkyl group contains between 1 and 13 halogens. The halogen may be fluoro.

In another embodiment,

may be selected from the group consisting of: hydrogen, halogen, phenyl, Ci-C₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens, and wherein the phenyl group may optionally be substituted with Ci-C₆ alkyl, Ci-C₆ alkoxy, hydroxy or halogen. The halogen may be fluoro. In a further embodiment, R| may be selected from the group consisting of: halogen, phenyl, C1-C4 alkyl and halo C1-C4 alkyl, wherein the halo C1-C4 alkyl group contains between 1 and 4 halogens, and wherein the phenyl group may optionally be substituted with Ci-C₆ alkyl, Ci-C₆ alkoxy or hydroxy. The halogen may be fluoro.

In an alternative embodiment, R_\ may be selected from the group consisting of: phenyl, C1-C4 alkyl and halo C1-C4 alkyl, wherein the halo C1-C4 alkyl group contains between 1 and 3 halogens, and wherein the phenyl group may optionally be substituted with Ci-C₆ alkyl, Ci-C₆ alkoxy or hydroxy. The halogen may be fluoro.

In still another embodiment, R_\ may be selected from the group consisting of: phenyl, C1-C4 alkyl and halo C1-C4 alkyl, wherein the halo C1-C4 alkyl group contains between 1 and 3 fluoro substituents, and wherein the phenyl group may optionally be substituted with Ci -C₆ alkyl, Cj -C₆ alkoxy or hydroxy.

In yet a further embodiment, Ri may be selected from the group consisting of: phenyl, C1-C4 alkyl and halo C1-C4 alkyl, wherein the halo C1-C4 alkyl group contains between 1 and 3 fluoro substituents, and wherein the phenyl group may optionally be substituted with one or more of the following substituents: methyl, hydroxy and methoxy.

In another embodiment, Ri may be selected from the group consisting of: phenyl, methyl, ethyl and propyl, wherein the methyl, ethyl and propyl groups may optionally be substituted with between 1 and 3 fluoro substituents, and wherein the phenyl group may optionally be substituted with one of the following substituents: methyl, hydroxy and methoxy.

In yet another embodiment, Ri may be selected from the group consisting of: phenyl, CH₃, CF₃, CHF₂, CH₂F, CH₂CHF₂, CH₂CH₂F and CH₂CF₃₎ and wherein the phenyl group may optionally be substituted with one of the following substituents: methyl, hydroxy arid methoxy.

In another embodiment, Ri may be optionally substituted aryl.

In yet a further embodiment, Ri may be phenyl, wherein the phenyl group may optionally be substituted with methyl, ethyl, methoxy, ethoxy, hydroxy and halogen.

In still a further embodiment, Rj may be selected from the group consisting of: hydrogen, optionally substituted phenyl, Ci-C₆ alkyl, perfluoro Ci-C₆ alkyl, Ci -C₆ alkoxy and perfluoro Ci-C₆ alkoxy. In yet another embodiment, Rj may be selected from the group consisting of: hydrogen, phenyl, methyl, methoxy and perfluoroCi-Gj alkyl.

R₂ may be selected from the group consisting of: hydrogen, optionally substituted aryl and Ci-C₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens.

In an alternative embodiment, R₂ may be selected from the group consisting of: hydrogen, optionally substituted phenyl and Ci-C₆ alkyl.

In a further embodiment, R₂ may be selected from the group consisting of: hydrogen and phenyl, wherein the phenyl group may optionally be substituted with C]-C₆ alkyl, Ci-C₆ alkoxy, hydroxy or halogen.

In still a further embodiment, R₂ may be selected from the group consisting of: hydrogen and phenyl, wherein the phenyl group may optionally be substituted with one or more of the following substituents: methyl, ethyl, methoxy, ethoxy, hydroxy and halogen.

R₃, R4, R₅, R-s and R₇ may be independently selected from the group consisting of: hydrogen, hydroxy, halogen, Ci-C₆ alkyl and Ci-C₆ alkoxy.

In another embodiment, R3, R4, R₅, R and R₇ may be independently selected from the group consisting of: hydrogen, hydroxy, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy and halogen.

In a further embodiment, R₃, R4, R5, ¾ and R₇ may be independently selected from the group consisting of: hydrogen, hydroxy, methyl, ethyl, methoxy, ethoxy and halogen.

In still another embodiment, R₃, R4, R₅, and R₇ may be independently selected from the group consisting of: hydrogen, methyl, ethyl, methoxy and halogen.

In a further embodiment, R₃, R4, R₅, R and R₇ may be independently selected from the group consisting of: hydrogen, methyl, ethyl, methoxy, chloro and bromo.

In yet another embodiment, R₃, R4, R5 and R₇ are hydrogen, and R₅ is selected from the group consisting of: hydroxy, C C6 alkyl, Ci-C₆ alkoxy and halogen.

In still a further embodiment, R₃, R4, Re and R₇ are hydrogen, and R₅ is selected from the group consisting of: hydrogen, hydroxy, methyl, ethyl, methoxy, ethoxy and halogen. ¾ and R₉ may be independently selected from the group consisting of: hydrogen, methyl, ethyl, propyl and isopropyl.

In one embodiment, X is O, Y is NRs, R_\ is selected from the group consisting of: hydrogen, halogen, optionally substituted aryl, optionally substituted heteroaryl, Ci-C₆ alkyl and halo C|-C₆ alkyl, wherein the halo C]-C₆ alkyl group contains between 1 and 6 halogens, R₂ is selected from the group consisting of: hydrogen, optionally, substituted aryl, Ci -C₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens, R₃, R4, R₅, R<s and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, halogen, Ci-C₆ alkyl and Ci-C₆ alkoxy, and R₈ is selected from the group consisting of: hydrogen, methyl, ethyl, propyl and isopropyl.

In another embodiment, X is 0, Y is NR₈, R_\ is selected from the group consisting of: hydrogen, halogen, optionally substituted aryl, C1-C6 alkyl and halo Ci-C₆ alkyl, wherein the halo C]-C₆ alkyl group contains between 1 and 6 halogens, R₂ is selected from the group consisting of: hydrogen, optionally substituted aryl, C1-C6 alkyl, halo Ci-C₆ alkyl, wherein the halo Ci-Q alkyl group contains between 1 and 6 halogens, R₃, R4, R5, R₆ and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, halogen, Ci-C₆ alkyl and Ci-C₆ alkoxy and R₈ is hydrogen or methyl.

In a further embodiment, X is O, Y is NR₈, Ri is selected from the group consisting of: hydrogen, halogen, phenyl, Ci-C₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 4 halogens, and wherein the phenyl group may optionally be substituted with one or more of the following substituents: halogen, hydroxy, Ci-C₆ alkyl and Ci-C₆ alkoxy, R₂ is selected from the group consisting of: hydrogen, phenyl, Ci-C₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens and wherein the phenyl group may optionally be substituted with one or more of the following substituents: halogen, hydroxy, C]-C₆ alkyl and Ci-C₆ alkoxy, R₃, R4, R , R_$ and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, halogen, Ci-C₆ alkyl and Ci-C₆ alkoxy and R₈ is hydrogen or methyl.

In still a further embodiment, X is O, Y is NR₈, Ri is selected from the group consisting of: hydrogen, halogen, phenyl, Ci-C₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens, and wherein the phenyl group may be optionally substituted with one or more of the following substituents: halogen, hydroxy, C1-C6 alkyl and Ci-C₆ alkoxy, R₂ is selected from the group consisting of: hydrogen, phenyl and Ci-C₆ alkyl, wherein the phenyl group may optionally be substituted with one or more of the following substituents: halogen, hydroxy, Ci-C₆ alkyl and Ci-C₆ alkoxy, R₃, R₄, R₅, R6 and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, halogen, Ci-C₆ alkyl and Ci-C₆ alkoxy, and R₈ is hydrogen or methyl.

In yet another embodiment, X is O, Y is NR₈, Ri is selected from the group consisting of: hydrogen, phenyl, Ci-C₆ alkyl and halo Ci-C₆alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 fluoro substituents, and wherein the phenyl group may be substituted with one or more of the following substituents: halogen, hydroxy, methyl, ethyl, propyl, methoxy, ethoxy and propoxy, R₂ is selected from the group consisting of: hydrogen, phenyl and Ci-C₆ alkyl, wherein the phenyl group may optionally be substituted with one or more of the following substituents: methyl, ethyl, hydroxy, methoxy and ethoxy, R₃, R₄, R5, ^ and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, halogen, methyl, ethyl, methoxy and ethoxy, and R₈ is hydrogen.

In yet another embodiment, X is O, Y is NR₈, Ri is selected from the group consisting of: hydrogen, phenyl, CH₃, CF₃, CHF₂, CH₂F, CH₂CHF₂, CH₂CH₂F and CH₂CF , wherein the phenyl group may optionally be substituted with one or more of the following substituents: halogen, hydroxy, methyl, ethyl, methoxy and ethoxy, R₂ is selected from the group consisting of: hydrogen and phenyl, wherein the phenyl group may optionally be substituted with one or more of the following substituents: methyl, ethyl, hydroxy, methoxy and ethoxy, R₃, R₄, R₅, and R7 are independently selected from the group consisting of: hydrogen, hydroxy, bromo, chloro, methyl, ethyl, methoxy and ethoxy, and wherein at least three of R₃, R₄, R₅, ¾ and R₇ are hydrogen, and R₈ is hydrogen.

In yet another embodiment, X is O, Y is NR₈, Ri is selected from the group consisting of: phenyl, CH₃, CF₃, CH₂CH₃ and CF₂CF₃, R₂ is selected from the group consisting of: hydrogen and phenyl, R₃, R4, R₅, R6 and R₇ are independently selected from the group consisting of: hydrogen, bromo and chloro, and R₈ is hydrogen.

In still a further embodiment X is O, Y is NRg, Ri is selected from the group consisting of: phenyl, CH₃, CF₃ and CF₂CF₃, R₂ is selected from the group consisting of: hydrogen and phenyl, R₃, R₄, R₅, R6 and R₇ are independently selected from the group consisting of: hydrogen, chloro and bromo, wherein at least three of R₃, R4, R₅, ¾ and R₇ are hydrogen, and Rg is hydrogen.

In a further embodiment, X is O, Y is NRg, Ri is selected from the group consisting of: optionally substituted aryl, R₂ is selected from the group consisting of: hydrogen, optionally substituted aryl, Ci-C₆ alkyl and halo C]-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens, R₃, R4, R5, R<5 and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, halogen, Ci-C₆ alkyl and CpC₆ alkoxy and Rg is selected from the group consisting of: hydrogen, methyl, ethyl, propyl and isopropyl.

In another embodiment, X is O, Y is NRg, Ri is selected from the group consisting of: optionally substituted phenyl, R₂ is selected from the group consisting of: hydrogen, optionally substituted aryl, C1-C6 alkyl and halo C1-C6 alkyl, wherein.the halo Ci-C₆ alkyl group contains between 1 and 6 halogens, R₃, R4, R₅, R6 and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, halogen, Ci-C₆ alkyl and Ci-C₆ alkoxy and Rg is selected from the group consisting of: hydrogen, methyl, ethyl, propyl and isopropyl.

In a further embodiment, X is O, Y is NRg, Ri is phenyl which may optionally be substituted with one or more of the following substituents: halogen, hydroxy, Ci-C₆ alkyl and Ci-C₆ alkoxy, R₂ is selected from the group consisting of: hydrogen, phenyl, C1-C6 alkyl and halo C1-C6 alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens and wherein the phenyl group may optionally be substituted with one or more of the following substituents: halogen, hydroxy, Ci-C₆ alkyl and Ci-C₆ alkoxy, R , R4, R5, ¾ and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, halogen, Ci-C₆ alkyl and Ci-C₆ alkoxy and Rg is hydrogen or methyl. '

In still a further embodiment, X is O, Y is NRg, Ri is phenyl which may optionally be substituted with one or more of the following substituents: halogen, hydroxy, methyl, ethyl, methoxy and ethoxy, R₂ is selected from the group consisting of: hydrogen, phenyl and Ci-C₆ alkyl, wherein the phenyl group may optionally be substituted with one or more of the following substituents: halogen, hydroxy, Ci-C₆ alkyl and Ci-C₆ alkoxy, R₃, R4, R₅, Re and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, halogen, Ci-C₆ alkyl and C1-C6 alkoxy, and Rg is hydrogen or methyl.

In yet another embodiment, X is O, Y is NRg, Ri is phenyl which may optionally be substituted with, one or more of the following substituents: halogen, hydroxy, methyl, ethyl, methoxy and ethoxy, R₂ is selected from the group consisting of: hydrogen, phenyl and Ci-C₆ alkyl, wherein the phenyl group may optionally be substituted with one or more of the following substituents: methyl, ethyl, hydroxy, methoxy and ethoxy, R₃, R4, R₅, R6 and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, halogen, methyl, ethyl, methoxy and ethoxy, and R₈ is hydrogen.

In yet another embodiment, X is O, Y is NR₈, Ri is phenyl which may optionally be substituted with one or more of the following substituents: hydroxy, methyl, ethyl, methoxy and ethoxy, R₂ is selected from the group consisting of: hydrogen and phenyl, wherein the phenyl group may optionally be substituted with one or more of the following substituents: methyl, ethyl, hydroxy, methoxy and ethoxy, R₃, R4, R₅, and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, bromo, chloro, methyl, ethyl, methoxy and ethoxy, and Rg is hydrogen.

The compounds of formula (I) may possess one or more chiral centres. The present invention includes all enantiomers and diastereoisomers, as well as mixtures thereof in any proportions. The invention also extends to isolated enantiomers or pairs of enantiomers. Enantiomers and diastereoisomers may be separated according to methods well known to those skilled in the art.

Synthesis of compounds of formula (I)

Compounds of the formula (I) may be conveniently prepared from appropriately substituted compounds of the formula (II) by reaction with an appropriately functionalised β-keto ester of the formula R|-C(0)CH₂C(0)-R as depicted in Scheme 1.

(Π) (I)

Scheme 1 - Preparation of compounds of the formula (I)

Compounds of the formula (II) may be prepared for example, by demethylating the corresponding 4,6-dimethoxy analogues. The corresponding 4,6-dimethoxy analogues may be prepared according to the methods described in Black, D. St C, Kumar, N., Wong, L. C. H., Aust. J. Chem., (1986), 39, 15-20, and Black, D. St C, Bowyer, M., Bowyer, P. K., Ivory, A. J., Kim, M., Kumar, N., McConnell, D. B., Aust. J. Chem., (1994), 47, 1741- 1750. Persons skilled in the art will understand that the above synthesis can be modified to use different starting materials and reagents in order to accomplish alternative desired transformations.

The present invention is predicated on the inventor's surprising finding that compounds of the formula (I) are capable of inhibiting the proliferation of cells, and hence are useful in the prevention and/or treatment of diseases and disorders characterised by aberrant cell proliferation, for example cancer. Examples of cancers that may be treated or prevented include, but are not limited to: gastric cancer, cancer of the liver and biliary tract, pancreatic cancer, prostate cancer (including androgen-dependent and androgen- independent), testicular cancer, lung cancer (including small cell and non-small cell), skin cancer (for example melanoma), breast cancer, non-melanoma skin cancer (for example basal cell carcinoma and squamous cell carcinoma), ovarian cancer, uterine cancer, cervical cancer, cancer of the head and neck, bladder cancer, sarcomas and osteosarcomas, Kaposi sarcoma, AIDS-related Kaposi sarcoma, renal carcinoma, leukaemia, colorectal cancer and glioma. The cancer may be a primary or secondary cancer. The cancer may be a non-androgen dependent cancer and/or a non-hormone dependent cancer.

In one embodiment, the methods and compositions of the present invention are particularly suitable for the treatment of the following cancers: liver cancers such as hepatocellular carcinomas, non-small cell lung cancer, breast cancer, gastric cancer and colorectal cancer.

In the treatment or prevention of cancer, therapeutic advantages may be obtained through combination treatment regimens. As such, methods of treatment of cancer according to the present invention may be used in conjunction with other therapies, such as radiotherapy, chemotherapy, surgery, or other forms of medical intervention. Non-limiting examples of suitable chemotherapeutic and other anti-cancer agents include: taxol, fluorouracil, cisplatin, oxaliplatin, cc-interferon, vincristine, vinblastine, angioinhibins, doxorubicin, bleomycin, mitomycin C, phenoxodiol, NV-128, NV-196, methramycin, TNP-470, pentosan polysulfate, tamoxifen, LM-609, CM- 101 and SU-101.

The co-administration of compounds of the formula (I) and chemotherapeutic or other anti-cancer agents may be simultaneous or sequential. Simultaneous administration may be effected by a compound of the formula (I) being in the same unit dose as a chemotherapeutic or other anti-cancer agent, or the compound of the formula (I) and the chemotherapeutic or other anti-cancer agents may be present in individual and discrete unit doses administered at the same, or at a similar time. Sequential administration may be in any order as required, and may require an ongoing physiological effect of the first or initial compound to be current when the second or later compound is administered, especially where a cumulative or synergistic effect is desired.

Pharmaceutical compositions and routes of administration

The compounds of formula (I) are useful as therapeutic agents in the treatment or prevention of various diseases and disorders associated with abberant cell proliferation. The compounds of formula (I) may be administered to a subject in the form of pharmaceutical compositions.

Pharmaceutical compositions include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous and intraarticular), inhalation (including use of metered dose pressurised aerosols, nebulisers or insufflators), rectal and topical (including dermal, buccal, sublingual and intraocular) administration. Generally, an effective dosage of a compound of the formula (I) is expected to be in the range of about O.OOOlmg to about lOOOmg per kg body weight per 24 hours; about O.OOlmg to about 750mg per kg body weight per 24 hours; about O.Olmg to about 500mg per kg body weight per 24 hours; about 0.1 mg to about 500mg per kg body weight per 24 hours; about O.lmg to about 250mg per kg body weight per 24 hours, or about l.Omg to about 250mg per kg body weight per 24 hours. More typically, an effective dose range is expected to be in the range of about l.Omg to about 200mg per kg body weight per 24 hours; about 1.Omg to about lOOmg per kg body weight per 24 hours; about 1.Omg to about 50mg per kg body weight per 24 hours; about 1.Omg to about 25mg per kg body weight per 24 hours; about 5.0mg to about 50mg per kg body weight per 24 hours; about 5. Omg to about 20mg per kg body weight per 24 hours, or about 5. Omg to about 15mg per kg body weight per 24 hours.

Alternatively, an effective dosage may be up to about 500mg/m². Generally, an effective dosage is expected to be in the range of about 25 to about 500mg/m , about 25 to about 350mg/m², about 25 to about 300mg m², about 25 to about 250mg/m², about 50. to about 250mg/m², or about 75 to about 150mg/m².

The therapeutically effective dose level of a compound of the present invention and its route of administration for any particular patient will typically depend upon a variety of factors including any one or more of: the type of cancer being treated and the stage of the cancer; the activity of the compound employed; the rate of metabolism and/or degradation of the compound; the age, body weight, general health, sex and diet of the patient; the time of administration; the rate of sequestration of compounds; the duration of the treatment; drugs used in combination or coincidental with the treatment, together with other related factors well known in medicine.

One skilled in the art would be able, by routine experimentation, to determine an effective, non-toxic dosage of a compound which would be required to treat applicable cancers, and also the most effective route of administration. These will most often be determined on a case-by-case basis.

Pharmaceutical compositions of the present invention may conveniently be prepared by methods well known in the art of pharmacy. All methods incl.ude the step of bringing one or more compounds of the formula (I) into association with one or more pharmaceutically acceptable carrier, diluent and/or excipient. In general, the compositions are prepared by uniformly and intimately bringing into association one or more compounds of the formula (I) with a liquid carrier or finely divided solid carrier.

Examples of pharmaceutically acceptable carriers, diluents and excipients include but are not limited to: demineralised or distilled water, saline solution, vegetable based oils such as peanut oil, safflower oil, olive oil, cottonseed oil, maize oil, sesame oils such as peanut oil, safflower oil, olive oil, cottonseed oil, maize oil, sesame oil, arachis oil or coconut oil; silicone oils, including polysiloxanes, such as methyl polysiloxane, phenyl polysiloxane and methylphenyl polysolpoxane; volatile silicones; mineral oils such as liquid paraffin, soft paraffin or squalane; cellulose derivatives such as methyl cellulose, ethyl cellulose, carboxymethylcellulose, sodium carboxymethylcellulose or hydroxypropylmethylcellulose; lower alkanols, for example ethanol or iso-propanol; lower aralkanols; lower polyalkylene glycols or lower alkylene glycols, for example polyethylene glycol, polypropylene glycol, ethylene glycol, propylene glycol, 1,3-butylene glycol or glycerin; fatty acid esters such as isopropyl palmitate, isopropyl myristate or ethyl oleate; polyvinylpyrridone; agar; carrageenan; gum tragacanth or gum acacia, and petroleum jelly. The diluent may be dextran, trehalose, carboxymethylcellulose, albumin, propylene glycol in 99% water. Typically the carriers, diluents and excipients will form from 10% to 99.9% by weight of the compositions. Carriers, diluents and excipients must, of course, be acceptable in the sense of being compatible with any other components of the composition and must not be deleterious to the recipient thereof.

Compositions suitable for oral administration may be presented as discrete units such as gelatine or HPMC capsules, cachets or tablets, each containing a predetermined amount of a compound of formula (I) as a powder, granules, as a. solution or suspension in an aqueous liquid or a non-aqueous liquid, or as an oil-in-water liquid emulsion or a water- in-oil liquid emulsion. The compound of formula (I) may also be present as a paste.

When compounds of the formula (I) are formulated as capsules, the compound may be formulated with one or more pharmaceutically acceptable carriers such as starch, lactose, microcrystalline cellulose, silicon dioxide and/or a cyclic oligosaccaride such as cyclodextrin. Additional ingredients may include lubricants such as magnesium stearate and/or calcium stearate. Suitable cyclodextrins include a-cyclodextrin, β-cyclodextrin, γ- cyclodextrin, 2-hydroxyethyl-p-cyclodextrin, 2-hydroxypropyl-cyclodextrin, 3- hydroxypropyl-P-cyclodextrin and tri-methyl-P-cyclodextrin. The cyclodextrin may be hydroxypropyl-p-cyclodextrin. Suitable derivatives of cyclodextrins include Captisol® a sulfobutyl ether derivative of cyclodextrin and analogues thereof as described in US patent No. 5,134,127.

Tablets may be prepared by compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the compound of formula (I) in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant (for example magnesium stearate or calcium stearate), inert diluent or a surface active/dispersing agent. Moulded tablets may be made by moulding a mixture of the powdered compound of formula (I) moistened with an inert liquid diluent, in a suitable machine. The tablets may optionally be coated, for example, with an enteric coating and may be formulated so as to provide slow or controlled release of the compound of formula (I) therein.

Compositions for parenteral administration include aqueous and non-aqueous sterile injectable solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient, and which may include suspending agents and thickening agents. A parenteral composition may comprise a cyclic oligosaccaride such as hydroxypropyl-p-cyclodextrin. The compositions may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example saline or water- for-injection, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

Dry powder compositions for topical delivery to the lung by inhalation may, for example, be presented in capsules and cartridges of, for example gelatine, or blisters or for example laminated aluminium foil, for use in an inhaler or insufflator. Compositions generally contain a powder mix for inhalation of the one or more compounds of the formula (I) and a suitable powder base (carrier substance) such as lactose or starch. Use of lactose is preferred. Each capsule or cartridge may generally contain between 20 g-10mg of a compound of formula (I), optionally in combination with another therapeutically active ingredient. Alternatively, the compound or compounds of the formula (I) may be presented without excipients. Packaging of the composition may be for unit dose or multi- dose delivery. Spray compositions for topical delivery to the lung by inhalation may, for example be formulated as aqueous solutions or suspensions or as aerosols, suspensions or solutions delivered from pressurised packs, such as a metered dose inhaler, with the use of a suitable liquefied propellant. Suitable propellants include a fliiorocarbon or a hydrogen-containing chlorofluorocarbon or mixtures thereof, particularly hydrofluoroalkanes, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, especially 1,1,1 ,2-tetrafluoroethane, 1 ,1 , 2,2,3, 3,3-heptafluoro-n-propane or a mixture thereof. Carbon dioxide or Other suitable gas may also be used as propellant. The aerosol composition may be excipient free or may optionally contain additional composition excipients well known in the art, such as surfactants e.g. oleic acid or lecithin and cosolvents e.g. ethanol. Pressurised compositions will generally be retained in a canister (e.g. an aluminium canister) closed with a valve (e.g. a metering valve) and fitted into an actuator provided with a mouthpiece.

Medicaments for administration by inhalation desirably have a controlled particle size. The optimum particle size^" for inhalation into the bronchial system is usually 1-10 μιη, preferably 2-5 μπι. Particles having a size above 20 μηι are generally too large when inhaled to reach the small airways. When the excipient is lactose it will typically be present as milled lactose, wherein not more than 85% of lactose particles will have a MMD of 60-90 μπι and not less than 15% will have a MMD of less than 15 μπι.

Compositions suitable for buccal (sublingual) administration include lozenges comprising a compound of the formula (I) in a flavoured base, usually sucrose and acacia or tragacanth; and pastilles comprising a compound of the formula (I) in an inert base such as gelatine and glycerin or sucrose and acacia.

Compositions suitable for transdermal administration may be presented as discrete patches adapted to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. Such patches suitably^' comprise the compound of the formula (I) as an optionally buffered aqueous solution of, for example, 0.1 M to 0.2 M concentration with respect to the compound.

Compositions suitable for transdermal administration may also be delivered by iontophoresis, and typically take the form of an optionally buffered aqueous solution of the active compound. Suitable compositions comprise citrate or Bis/Tris buffer (pH 6) or ethanol/water and contain from 0.1 M to 0.2 M of a compound of the formula (I). Compositions for rectal administration may be presented as a suppository with carriers such as cocoa butter or polyethylene glycol, or as an enema wherein the carrier is an isotonic liquid such as saline. Additional components of the compositions may include a cyclic oligosaccaride, for example, a cyclodextrin, . as described above, such as hydroxypropyl-β- cyclodextrin, one or more surfactants, buffer salts or acid or alkali to adjust the pH, isotonicity adjusting agents and/or anti-oxidants.

Compositions suitable for topical administration to the skin preferably take the form of an ointment, cream, lotion, liniment, paste, gel, spray, aerosol or oil. Carriers which may be used include Vasoline, lanoline, polyethylene glycols, alcohols and combinations thereof. The compound of the formula (I) is generally present at a concentration of from 0.1% to 5% w/w, or from 0.5% to 2% w/w. Examples of such compositions include cosmetic skin creams. Lotions or liniments for application to the skin may also include an agent to hasten drying'and to cool the skin, such as an alcohol or acetone, and/or a moisturiser such as glycerol, or oil such as castor oil or arachis oil.

The compounds of the formula (I) may be provided in the form of food stuffs, such as being added to, admixed into, coated, combined or otherwise added to a food stuff. The term "food stuff is used in its widest possible sense and includes liquid compositions such as drinks, including dairy products and other foods, such as health bars, desserts, etc. Food compositions comprising compounds of the formula (I) can be readily prepared according to standard practices.

The composition may also be administered or delivered to target cells in the form of liposomes. Liposomes are generally derived from phospholipids or other lipid substances, and are formed by mono- or multi-lamellar hydrated liquid crystals that are dispersed in an aqueous medium. Specific examples of liposomes used in administering or delivering a composition to target cells are synthetic cholesterol (Sigma), 1 ,2-distearoyl- 5«-glycero-3-phosphocholine (DSPC; Avanti Polar Lipids), 3-N-[(-methoxy poly(ethylene glycol)2000)carbamoyl]-l,2-dimyrestyloxy-propylamine (PEG-cDMA), or 1 ,2-di-o- octadecenyl-3-(N,N-dimethyl)aminopropane (DODMA).

The compositions may also be administered in the form of microparticles. Biodegradable microparticles formed from polylactide (PLA), polylactide-co-glycolide (PLGA), and έ-caprolactone have been extensively used as drug carriers to increase plasma half life and thereby prolong efficacy (R. Kumar, M., 2000, J. Pharm. Pharmaceut. Sci. 3(2) 234-258). .

The compositions may incorporate a controlled release matrix that is composed of sucrose acetate isobutyrate (SAIB) and organic solvent or organic solvent mixtures. Polymer additives may be added to the vehicle as a release modifier to further increase the viscosity and slow down the release rate. A compound of the formula (I) may be added to the SAIB delivery vehicle to form SAIB solution or suspension compositions. When the formulation is injected subcutaneously, the solvent diffuses from the matrix allowing the SAIB-drug or SAIB-drug-polymer mixtures to set up as an in situ forming depot.

The present invention will now be described with reference to specific examples, which should not be construed as in any way limiting the scope of the invention.

Examples

Example 1 - Preparation of compounds

Synthesis of 3-(4-chlorophenyl)-lH-indole-4,6-dioI

A mixture of 3-(4-chlorophenyl)-4,6-dimethoxyindole (0.274 g, 0.93 mmol) and anhydrous aluminium chloride (0.320 g, 2.40 mmol) in sodium dried toluene (5 mL) was heated under reflux at 150 °C for 1.5 h. The reaction mixture was added to ice (20 mL) and extracted with ethyl acetate (2 x 10 mL), the extract washed with 10% hydrochloric acid (2 x 10 mL) and water (2 x 10 mL). The organic phase was dried over anhydrous magnesium sulfate and concentrated under reduced pressure to yield 3-(4-chlorophenyl)indole-4,6-diol as a beige solid (0.209 g, 85 %).

M.p. 178-180 °C (from hexane).

Found: C, 65.1 1 ; H, 3.99; N, 5.16%_r C₁₄Hi₀ClNO₂O.05hexane requires C, 65.06; H, 4.09; N, 5.31%.

Ή NMR (300 MHz, d₆-DMSO): δ 5.96 (d, J 1.5 Hz, 1H, H5), 6.24 (d, J 0.75 Hz, 1H, H7), 6.78 (d, J 1.1 Hz, 1H, H2), 7.41 (d, J 8.6 Hz, 2H, ArH), 7.71 (d; J 8.3 Hz, 2H, ArH), 8.81 (s, 1H, OH), 9.33 (s, 1H, OH), 10.98 (br s, 1H, NH).

¹³C NMR (75 MHz, d^-DMSO): δ 88.44 (ArC7), 95.39 (ArC5), 97.50 (ArC2), 1 13.03 (C), 125.94 (2 x ArC), 129.10 (2 x ArC), 130.41 (C), 132.08 (C), 132.75 (C), 140.06 (C), 151.24 (C), 155.18 (C). * IR (KBr): u_max 3371 , 1634, 1596, 1520, 1487, 1465, 1369, 1264, 1211, 1 150, 1078, 1010, 984, 833, 803, 783, 736, 663, 619 cm^"'.

UV (MeOH): _max 214 nm, (e 30706 cm^'1 M^"1), 259, (21580), 333, (22996).

HRMS (negative E.S.I.): Found m/z 258.0327, [M-H]^'; Ci₄H₉ClN0₂ requires 258.0321.

Synthesis of 2,3-diphenyl-lH-indoIe-4,6-diol

Anhydrous aluminium chloride (2.02 g, 15.17 mmol) was added portionwise at room temperature to a suspension of 4,6-dimethoxy-2,3-diphenyl indole (1.00 g, 3.03 mmol). The mixture was stirred at room temperature for 15 mins. followed by heating at 140° C for 1 hr. When a TLC showed that the starting material had been consumed the reaction mixture was allowed to cool to room temperature and poured onto crushed ice. The resulting white precipitate was filtered and washed with water until the washings were neutral. The solid was dried under vacuum to give 0.8 g of 2,3-diphenyl-lH-indole-4,6- diol.

M.p. 265 °C dec (from hexane).

Ή NMR (300 MHz, d^-DMSO): δ 6.12 (d, J 1.9 Hz, 1H, H5), 6.46 (d, J 1.9 Hz, 1H, H7), 7.12 - 7.42 (m, 10H, Ph), 7.55 (br s, 1H, OH), 7.87 (br s, 1H, OH), 10.15 (br s, 1H, NH). ¹³C NMR (75 MHz, d₆-DMSO): 88.46 (ArC7), 95.47 (ArC5), 111.62 (C), 1 13.93 (C), 125.78 (ArC), 126.31 (ArC), 127.43 (2 x ArC), 127.56 (2 x ArC), 128.05 (2 x ArC), 131.25 (2 x ArC), 133.35 (C), 136.87 (C), 139.08 (C), 151.92 (C), 154.65(C). IR (KBr): Umax 3274, 1634, 1507, 1451, 1382, 1238, 1115, 696 cm^'1.

UV (MeOH): λ™_* 204 nm, (ε16280 cm^'1 M^"1), 241 (8435), 291 (7815), 368 (2939). HRMS (ESI): Found m/z 324.0995 , [M+Na] ; C₂₀H, ₅N0₂Na requires 324.1000.

Synthesis of 4-hydroxy-2,3-diphenyl-9-(trifluoromethyl)pyrano[2,3-g]indol-7(lH)-

absolute ethanol was refluxed or 24 rs. n coo ng, the

resulting yellow solid was filtered, washed with water and dried to give 0.25 g of 4- hydroxy-2,3-diphenyl-9-(trifluoromethyl)pyrano[2,3-g]indol-7(lH)-one. Ή NMR (300 MHz, d₆-DMSO): δ 5.87 (bs, 1H, OH), 6.68 (2s, 2H, H5 and H8), 7.20-7.30 (m, 5H, ArH), 7.47-7.48 (m, 5H, ArH), 8.98 (bs, 1H, NH).

Synthesis of 3-(4-chlorophenyl)-4-hydroxy-9-phenylpyrano[2,3-g]indol-7(lH)-one.

A mixture of 3-(4-chlorophenyl)-lH-indole-4,6-diol (0.3g,

1.15 mmol), ethyl 3-oxo-3-phenylpropanoate 3.5 ml (18.53

mmol) and zinc chloride (0.189 g, 1.38 mmol) in 50 ml of

absolute ethanol was refluxed for 24 hrs. The resulting

yellow solid was filtered, washed with water and dried to

give 0.15 g of 3-(4-chlorophenyl)-4-hydroxy-9-

phenylpyrano[2,3-g]indol-7(lH)-one.

Ή NMR (300 MHz, c^-DMSO): δ 6.14 (s, 1H, H8), 6.63 (s, 1H, H5), 7.08 (d, 1H, H2), 7.2 (d, 2H, ArH), 7.4 (d, 2H, ArH), 7.6-7.8 (m, 5H, ArH), 8.00 (bs, 1H, NH).

Synthesis of 4-hydroxy-9-methyl-2,3-diphenylpyrano[2,3-g]indol-7 lH -one.

A mixture of 2,3-diphenyl-lH-indole-4,6-diol 0.3 g (0.995

mmol), ethyl 3-oxobutanoate 2.33 ml (15.0 mmol) and zinc

chloride 0.19 g (1.40 mmol) in 50 ml of absolute ethanol was

refluxed for 24 hrs. On cooling, the resulting white solid was

filtered, washed with water and dried to give 0.16 g of 4-hydroxy-9-methyl-2,3- diphenylpyrano[2,3-g]indol-7(lH)-one. Ή NMR (300 MHz, ^-DMSO): δ 2.84 (s, 3H, CH₃), 6.01 (s, 1H, H8), 6.43 (s, 1H, H5), 7.22 - 7.32 (m, 10H, Ph), 10.40 (br s, 1H, NH), 10.59 (br s, 1H, NH). IR (KBr): u_max 3474, 1685, 1623, 1446, 1382, 1102, 968, 800 cm^"1. UV (MeOH): λ™_χ 203 nm, (ε8343 cm^"1 M^"1), 250 (7350), 309 (61 18).

Example 2 - Anti-cancer activity

1.0 Tissue Culture

The human non-small cell lung adenocarcinoma lines NCI-H1299 (CRL-5803), NCI-H460 (HTB-177), NCI-H358 (CRL-5807), NCI-H838 (CRL-5844), the human colorectal adenocarcinoma lines COLO 205 (CCL-222), HCT-15 (CCL-225) and the human gastric cancer cell line NCI-N87 (CRL-5822) were cultured in RPMI 1640 medium containing 2g/L sodium bicarbonate (Hyclone/Invitrogen) supplemented with 2mM L- glutamine (Gibco) ImM sodium pyruvate (Sigma), lOmM HEPES (Sigma) and 4500mg/L glucose (Sigma),

The human non-small cell lung adenocarcinoma line NCI-H2126 (CCL-256) was cultured in DMEM:F12(1 :1) (Hyclone/Invitrogen) containing 2.5mM L-glutamine 2.4g/L sodium bicarbonate supplemented with 5% FBS, an additional 2mM L-glutamine, 15mM HEPES, 0.005mg/ml insulin (Sigma), O.Olmg/ml transferrin (Sigma), 30nM sodium selenite (Sigma), lOnM hydrocortisone (Sigma) and ΙΟηΜ beta-estradiol (Sigma).

The human colorectal adenocarcinoma line HT-29 (HTB-38) and the human gastric cancer cell lines OE-19 (#96071721) and MKN1 (JCRB0252) were cultured in RPMI 1640 media supplemented with 2mM L-glutamine.

The human colorectal adenocarcinoma line HCT-1 16 (CCL-247) and the human breast adenocarcinoma line SK-BR-3 (HTB-30) were cultured in McCoy's 5a Medium (Invitrogen) containing 1.5mM L-glutamine and 2.2g/L sodium bicarbonate.

The human colorectal adenocarcinoma line SW620 (CCL-227) was cultured in Leibovitz's L-15 Medium (Invitrogen) containing 2.05mM L-glutamine.

The human hepatocellular carcinoma lines HepG2 (HB-8065), SK-HEP-1 (HTB- 52) and the normal human lung fibroblast line IMR-90 (CCL-186) were cultured in Minimum Essential Eagles Medium containing 2mM L-glutamine, 2.2g/L sodium bicarbonate, supplemented with ImM sodium pyruvate and O.lmM non-essential amino acids.

The human hepatocellular carcinoma line JHH-1 (JCRB1062) was cultured in Williams E Medium (Invitrogen) containing 2.2g/L sodium bicarbonate, supplemented with 2mM L-glutamine. The hepatocellular carcinoma line HuH-7 (JCRB0403) was cultured in DMEM, supplemented with 2mM L-glutamine.

The human gastric cancer cell line AGS (CRL-1739) was cultured in Hams F-12K (Kaighans modification) medium containing 2mM L-glutamine, 2.5g/L sodium bicarbonate and 2mM sodium pyruvate.

The human breast adenocarcinoma line MDA-MB-468 (HTB-132) was cultured in DMEM:F12 (1 : 1) medium containing 2.5mM L-glutamine, 2.4g/L sodium bicarbonate, supplemented with an additional 2mM L-glutamine. All cultures were supplemented with 10% FBS (unless stated otherwise), penicillin (lOOU/ml) and streptomycin (100μ¾/πι1) and cultured at 37°C in a humidified atmosphere of 5% C0_2, with the exception of SW-620 which was cultured at 37°C in standard humidified atmosphere_^ All cell lines were purchased from ATCC (Maryland, USA) with the exception of MKNl, HuH-7, JHH-1, purchased from the Japanese Collection of Research Bioresources (JGRB, Osaka, Japan), OE19 purchased from the European Collection of Cell Cultures (ECACC, Salisbury, UK) and R-90 was purchased from ATCC via Robin Anderson (Peter Mac Cancer Institute, Melbourne, Australia).

2.0 Proliferation Assays

IC50 and IC90 values were determined for each cell line. Cells were seeded in 96- well plates at an appropriate cell density as determined from growth kinetics analysis and cultured for 5 days in the absence and presence of the test compounds.

Cell proliferation was assayed following the addition of either:

2.1 20μΙ_, of a 5mg/ml MTT [3-4, 5 dimethylthiazol-2,5-diphenyl tetrazolium bromide] (Sigma) solution in PBS for 3-5 hours, or

2.2 20μί of CellTitre 96® AQ_ueous Non-Radioactive Cell Proliferation Assay (Promega), comprised of a MTS PMS solution (95.24% 3-(4,5- dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H- tetrazolium, 4.76% phenazine methosulfate)

Cells that grew partially in suspension (COLO 205), or did not reduce MTT to provide a control OD absorbance reading of greater than 0.4 (NCI-H358) were assayed using the CellTitre 96® AQ_ue0us Non-Radioactive Cell Proliferation Assay.

Cells were incubated under appropriate conditions for 3-5 hours (MTT) or 2-3 hours (MTS) and monitored for colour development prior to assay using a Molecular Devices, SpectraMAX 250.

MTT assays were assayed at 570nM whilst MTS assays were assayed at 440nM less a background of 690nM. ICso and IC90 values were calculation from semi-log plots of % of control proliferation on the y-axis against log-dose on the x-axis, using a simple linear interpolation of data points above/below the 50% and 10% levels of raw cell viability.

The results are shown in Table 1 below.

Table 1 : Effect of compounds 1 and 2 on inhibition of various cancer cell lines

Claims

CLAIMS:

1. A compound of the general formula (I):

wherein:

Y is O, S or NR₈

Ri is selected from the group consisting of: hydrogen, hydroxy, halogen, optionally substituted aryl, optionally substituted heteroaryl, alkylaryl, C(0)OCi-C₆ alkyl, Ci-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halo d⁽-C₆ alkyl, halo C₂-C₆ alkenyl, halo C₂-C₆ alkynyl, Ci-C₆ alkoxy and halo Ci-C₆ alkoxy,

R₂ is selected from the group consisting of: hydrogen, optionally substituted aryl, optionally substituted heteroaryl, Ci-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halo Ci-C₆ alkyl, halo C₂-C₆ alkenyl, halo C₂-C₆ alkynyl, Ci-C₆ alkoxy and halo Ci-C₆ alkoxy,

R₃, R4, R₅, R6 and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, Ci-C₆ alkoxy, amino, halogen, Ci-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, halo Ci-C₆ alkyl, halo C₂-C₆ alkenyl, halo C₂-C₆ alkynyl and halo C|-C₆ alkoxy,

Rs and R9 are independently selected from the group consisting of: hydrogen, Ci-C₆ alkyl and alkylaryl, j

the drawing "— " represents either a single bond or a double bond, and salts and derivatives thereof.

2. The compound of claim 1, wherein X is O.

3. The compound of claim 1 or claim 2, wherein Y is NR₈.

4. The compound of any one of claims 1 to 3, wherein Ri is selected from the group consisting of: hydrogen, halogen, optionally substituted aryl, optionally substituted heteroaryl, Ci-C₆ alkyl, halo Ci-C₆ alkyl, Ci-C₆ alkoxy and halo Ci-C₆ alkoxy.

5. The compound of any one of claims 1 to 4, wherein R] is selected from the group consisting of: hydrogen, halogen, optionally substituted aryl, optionally substituted heteroaryl, Cj-C₆ alkyl and halo C] -C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 13 halogens.

6. The compound of claim 5, wherein Ri is selected from the group consisting of: hydrogen, halogen, optionally substituted aryl, Ci -C₆ alkyl and halo Ci -C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 13 halogens.

7. The compound of claim 6, wherein Ri is selected from the group consisting of: hydrogen, halogen, phenyl, Ci-C₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens, and wherein the phenyl group may optionally be substituted with Q-C6 alkyl, Ci-C₆ alkoxy, hydroxy or halogen.

8. The compound of claim 7, wherein Ri is selected from the group consisting of: halogen, phenyl, C1 -C4 alkyl and halo C1-C4 alkyl, wherein the C1 -C4 alkyl group contains between 1 and 4 halogens, and wherein the phenyl group may optionally be substituted with Ci-C₆ alkyl, Ci-C₆ alkoxy or hydroxy.

9. The compound of any one of claims 1 to 8, wherein R₂ is selected from the group consisting of: hydrogen, optionally substituted aryl, Ci-C₆ alkyl and halo Ci -C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens.

10. The compound of claim 9, wherein R₂ is selected from the group consisting of: hydrogen, optionally substituted phenyl, Ci-C₆ alkyl and halo Ci-C₆ alkyl, wherein the halo Ci-C₆ alkyl group contains between 1 and 6 halogens.

11. The compound of claim 10, wherein R₂ is selected from the group consisting of: hydrogen, optionally substituted phenyl and Ci-C₆ alkyl.

12. The compound of any one of claims 1 to 1 1 , wherein R₃, R4, R₅, R¾ and R7 are independently selected from the group consisting of: hydrogen, hydroxy, halogen, Q- C₆ alkyl and Ci-C₆ alkoxy.

13. The compound of claim 12, wherein R₃, R4, R5, ¾ and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy and halogen.

14. The compound of claim 13, R₃, R4, R₅, R<5 and R₇ are independently selected from the group consisting of: hydrogen, hydroxy, methyl, ethyl, methoxy, ethoxy and halogen.

15. The compound of any one of claims 1 to 14, wherein R₈ and R are independently selected from the group consisting of: hydrogen, methyl, ethyl, propyl and isopropyl.

16. A compound of formula (I) according to claim 1, which is selected from compounds 1, 2 and 3.

17. A pharmaceutical composition comprising a compound of the formula (I) as defined in any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent and/or excipient.

18. A method for inhibiting the proliferation of cells, said method comprising contacting the cells with a compound of the formula (I), as defined in any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof.

19. A method for the prevention and/or treatment of cancer in a subject in need thereof, said method comprising administration to the subject of a therapeutically effective amount of a compound of the formula (I), as defined in any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof.

20. The method of claim 19, wherein the cancer is selected from the group consisting of: breast cancer, ovarian cancer, leukaemia, prostate cancer, liver cancer, colorectal cancer, gastric cancer, pancreatic cancer, glioma, melanoma and lung cancer.

21. The method of claim 20, wherein the cancer is selected from the group consisting of: breast cancer, ovarian cancer, leukaemia, non-androgen dependent prostate cancer, liver cancer, colorectal cancer, gastric cancer, pancreatic cancer, glioma, melanoma and lung cancer.

22. The method of claim 21, wherein the cancer is selected from the group consisting of: colorectal cancer, gastric cancer, liver cancer and lung cancer.

23. The method of claim 19, wherein the cancer is a non-androgen dependent cancer.

24. The method of claim 19, wherein the cancer is a non-hormone dependent cancer.