CROSS-REFERENCE TO RELATED PATENT APPLICATIONS This application claims priority to U.S. Provisional Application No. 61/287,340, filed December 17, 2009, which is incorporated herein by reference in its entirety.

FIELD

The present invention relates to substituted tetrahydro- -carbolines and substituted tetrahydro-y-carbolines. This invention also relates to methods of making these compounds. The compounds of this invention are inhibitors of Eg5. Therefore, the compounds of this invention are useful as pharmaceutical agents, especially in the treatment and/or prevention of a variety of diseases including diseases associated with cellular proliferation, such as cancer.

BACKGROUND

Antimitotic agents are an important category of cancer therapeutic drugs. These drugs are typically tubulin binders that block cell cycle progression by disrupting the normal assembly or disassembly of the mitotic spindle. However, antimicrotubule agents exhibit side effects, such as neutropenia and peripheral neuropathy.

The mitotic kinesin Eg5, also known as kinesin spindle protein (KSP), is required for bipolar spindle formation in the early phase of mitosis. Inhibition of Eg5 leads to cell cycle arrest, growth impairment and apoptosis induction in cancer cells. As such, Eg5 inhibitors can modulate cancer cell growth and survival in vivo. Furthermore, being mechanistically distinct from anti-microtubule agents, Eg5 inhibitors have not shown peripheral neuropathic side effects and thus may offer better quality of patient care. Accordingly, Eg5 is important as a target molecule of a new anticancer treatment, and it is considered that an inhibitor against said molecule is promising as an agent for therapeutic treatment of diseases caused by abnormality of the regulation of cell proliferation. SUMMARY

Thus, there are provided compounds of the formula (I):

wherein:

W represents CR6R7, S, or O;

X represents a nitrogen atom and Y represents CRs; or X represents CR9 and Y represents a nitrogen atom;

Ri , R2, R3, R4, R5, Re, and R ₉ , are the same or different and independently selected from the group consisting of hydrogen, halogen, cyano, amino, monoalkylamino, dialkylamino, cyanoalkyl, carbamide, haloalkyl, alkoxyalkyl, alkylcarbonyl, an unsubstituted or substituted alkyl, an unsubstituted or substituted alkoxy, an unsubstituted or substituted thioalkyi, and an unsubstituted or substituted aminoalkyi; wherein said alkyl, alkoxy, thioalkyi or aminoalkyi substituents are one or more radicals independently selected from the group consisting of alkyl, alkoxy, thioalkyi, and aminoalkyi;

R6 is selected from the group consisting of halogen, cyano, cyanoalkyl, haloalkyl, benzyl, -NR _a R _b , alkylcarbonyl, alkoxycarbonyl, arylcarbonyl, heteroarylcarbonyl, an unsubstituted or substituted aryl, an unsubstituted or substituted heteroaryl, an unsubstituted or substituted alkyl, an unsubstituted or substituted alkoxy, an unsubstituted or substituted thioalkyi, and an unsubstituted or substituted aminoalkyi; wherein when said aryl, heteroaryl, alkyl, alkoxy, thioalkyi or aminoalkyi represented by R6 is substituted, the substituents are one or more radicals independently selected from the group consisting of alkyl, halogen, phenyl, alkoxy, thioalkyi, and aminoalkyi;

R ₇ is selected from the group consisting of hydrogen, alkyl, cyano, and halogen;

or R ₃ and R6, together with the carbon atoms to which they are attached, form a cycloalkyl group; and R _a and R _b are the same or different and independently selected from the group consisting of hydrogen, alkyl, arylalkoxycarbonyl, and arylalkyl;

or a salt thereof.

In addition, various embodiments of this invention including pharmaceutical compositions comprising various compounds of this invention as well as their use in the treatment of a variety of disorders and/or disease conditions as disclosed herein are also part of this invention all of which are described in detail below.

DETAILED DESCRIPTION

Definitions and Abbreviations

As used above, and throughout the description, the following abbreviations, unless otherwise indicated, shall be understood to have the following meanings:

AcN acetonitrile

AcOH acetic acid

CDCIs deuterated chloroform

DABCO 1 , 4-diazabicyclo[2. 2. 2]octane solution

DCE 1 ,2-dichloroethane

DCM dichloromethane

DMA N, Ν-ό i m ethy I -aceta m id e

DME 1 ,2-dimethoxyethane

DMF N, Ν-ό i methylforma m ide

DMSO dimethyl sulfoxide

Et ₂ O diethyl ether

EtOAc ethyl acetate

EtOH ethanol

Et ₃ N triethylamine

FA formic acid

HPLC high performance liquid chromatography

Hunig's base diisopropylethylamine

□HMDS lithium bis(trimethylsilyl)amide

MeOH methanol

min minute MTBE terf-butyl methyl ether

NaHMDS sodium bis(trimethylsilyl)amide

Pd2(dba)3CHCl3 tris(dibenzylideneacetone)dipalladium(0)-chloroform adduct PdCl2(dppf).CH ₂ Cl2 1 ,1 '-bis(diphenylphosphino)ferrocene-palladium(ll)dichloride

dichloromethane complex

PPTS pyridinium p-toluenesulfonate

Py pyridine

t-BuOH tert-butanol

t-BuXPhos 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl

Tf trifluoromethanesulfonyl

TFA trifluoroacetic acid

THF tetrahydrofuran

As used above, and throughout the description of the invention, the following terms, unless otherwise indicated, shall be understood to have the following meanings:

As used herein, the term "compound(s) of the invention" means, collectively, the compounds of formula (I), pharmaceutically acceptable salts, solvates, or hydrates thereof, and mixtures thereof.

As used herein, the term "alkyl" means a straight or branched aliphatic hydrocarbon group having 1 to about 12 carbon atoms in the chain. In one aspect, an alkyl has 1 to about 6 carbon atoms in the chain. "Lower alkyl" means an alkyl group having 1 to about 4 carbon atoms in a linear alkyl chain that may be straight or branched. Branched means that one or more lower alkyl groups, such as methyl, ethyl or propyl, are attached to a linear alkyl chain. Also, "alk" denotes an alkyl group, unless divalency is indicated, in which case the "alk" denotes the corresponding alkylene group(s). Additionally, the term "alkyl _(C i-c6)" denotes an alkyl group having one to six carbon atoms. The term "alkyl ₍ ci-c4)" denotes an alkyl group having one to four carbon atoms. Exemplary alkyl includes methyl, ethyl, / ^' -propyl, f-butyl, and the like. As used herein, the term "alkoxy" means an alkyl-O- group wherein the alkyl group is as herein described. Exemplary alkoxy groups include methoxy, ethoxy, n-propoxy, / ^' -propoxy, n-butoxy, f-butoxy, heptoxy, and the like. As used herein, the term "alkoxyalkyl" means an alkyl-O-alkylene- wherein the alkyl and the alkylene are as herein described. Exemplary alkoxyalkyl groups include methoxymethylene and ethoxymethylene.

As used herein, the term "alkylcarbonyl" means a radical or substituent of the formula alkyl-C(=O)-, wherein the alkyl is as herein described. Exemplary alkylcarbonyl moieties include methylcarbonyl, ethylcarbonyl, propylcarbonyl, isopropylcarbonyl, butylcarbonyl, isobutylcarbonyl, t-butylcarbonyl, and pentylcarbonyl.

As used herein, the term "alkoxycarbonyl" means an alkyl-O-C(=O)- group, wherein the alkyl group is as herein defined. Exemplary alkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl, and f-butyloxycarbonyl, and the like.

As used herein, the term "alkylene" means a straight or branched bivalent hydrocarbon chain having from 1 to about 12 carbon atoms. In one aspect, an alkylene has 1 to about 10 carbon atoms in the chain. In another aspect, an alkylene has 1 to about 6 carbon atoms in the chain. A "lower alkylene" is an alkylene having from 1 to about 4 carbon atoms. Exemplary alkenylene includes methylene, ethylene, propylene, and butylene. As used herein, the term "aminoalkyi" means a NH ₂ -alkylene- wherein the alkylene is as herein described.

As used herein, the term "aryl", whether used alone or with other terms, such as "aryl group", "aryl radical", "aryl ring", and the like, means an aromatic monocyclic or multicyclic ring system of 6 to about 14 carbon atoms. In one aspect, aryl includes 6 to about 10 carbon atoms. Exemplary aryl include phenyl and naphthyl. As used herein, the term "arylalkyl" means aryl-alkylene- in which the aryl and alkylene moieties are as previously described. In one aspect, an arylalkyl is a aryl- alkyl(ci-c6)- moiety. Exemplary arylalkyl includes benzyl, 2-phenethyl and 1 - or 2- naphthalenemethyl.

As used herein, the term "arylalkoxycarbonyl" means aryl-alkoxy-C(=O)- in which the aryl and alkoxy groups are as previously described. Exemplary arylalkoxycarbonyl includes phenylmethoxycarbonyl. As used herein, the term "arylcarbonyl" means aryl-C(=O)- in which the aryl group is as previously described. Exemplary arylcarbonyl includes phenylcarbonyl.

As used herein, the term "benzyl" means phenyl-CH ₂ -. As used herein, the term "cancer" in a patient refers to the presence of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and certain characteristic morphological features. Cancer cells may be in the form of a tumor; may exist alone within a patient; may circulate in the blood stream as independent cells, such as leukemic cells; or be in the form of cancer cells with stem- cell like properties, such as leukemic stem cells.

As used herein, the term "carbamide" means a radical of the formula R'R"N-C(=O)- where R' and R" are independently hydrogen, alkyl, or cycloalkyl, as defined herein.

As used herein, the term "carbonyl," whether used alone or with other terms, such as "alkoxycarbonyl", means a -C(=O)- radical.

As used herein, the term "cyanoalkyl" means a radical or substituent of the formula - alkyl-C≡N, wherein the alkyl is as herein described.

As used herein, the term "cycloalkyl" means a non-aromatic monocyclic saturated ring system of 3 to about 10 carbon atoms, preferably of 3 to about 6 carbon atoms. Preferred ring systems include about 5 to about 6 ring atoms. Exemplary cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl, and the like.

As used herein, the term "dialkylamino" means (alkyl) ₂ -N- , wherein the alkyl groups, which may be the same or different, are as herein defined. Particular dialkylamino groups are (alkyl _(C i -c6))2-N-, wherein the alkyl groups may be the same or different. Exemplary dialkylamino groups include dimethylamino, diethylamino and methylethylamino.

As used herein, the terms "halo" or "halogen" mean fluoro, chloro, bromo, or iodo. Particular halogens are fluoro and chloro.

As used herein, the term "haloalkyi" means an alkyl substituted by one or more halo groups. Particular haloalkyi groups are lower alkyl substituted by one to three halogens. Exemplary haloalkyi include -CF ₃ , -CH ₂ CHF ₂ , and -CF ₂ CF ₃ .

As used herein, the term "heteroaryl" whether used alone or with other terms, such as "heteroaryl group", "heteroaryl radical", "heteroaryl ring", and the like, means a 5- to 10-membered aromatic monocyclic or multicyclic group having at least one carbon atom and at least one hetero element(s) other than carbon, for example nitrogen, oxygen or sulfur. The designation of the aza, oxa or thia as a prefix before heteroaryl defines that at least a nitrogen, oxygen or sulfur atom is present, respectively, as a ring atom. A nitrogen atom of a heteroaryl may be a basic nitrogen atom and may also be optionally oxidized to the corresponding N-oxide. When a heteroaryl is substituted by a hydroxy group, it also includes its corresponding tautomer where such hydroxy substituted heteroaryl is capable of such. Exemplary heteroaryl includes pyrazinyl, thienyl, isothiazolyl, oxazolyl, pyrazolyl, furazanyl, pyrrolyl, 1 ,2,4- thiadiazolyl, pyridazinyl, 1 ,2,4-triazinyl, furanyl, imidazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, pyrazinyl, pyridazinyl, pyrazolyl, pyridyl, pyrimidinyl, pyrrolyl, 1 ,3,4- thiadiazolyl, thiazolyl, thienyl, triazolyl, indolyl, quinolinyl, and the like.

As used herein, the term "monoalkylamino" means a radical of the formula alkyl-NH-, wherein the alkyl group is as herein defined. In one aspect, a monoalkylamino is an alkyl ₍ ci-c6 ₎ -arnino-. Exemplary monoalkylamino groups include methylamino and ethylamino.

As used herein, the term "patient" includes human and other mammals.

As used herein, the term "thioalkyl" means an S-alkyl- group in which the alkyl group is as previously described. Exemplary thioalkyl includes S-CH ₃ -.

As used herein, "treating", unless otherwise indicated, means to alleviate symptoms, eliminate the causation of the symptoms either on a temporary or permanent basis, or to slow the appearance of symptoms of the named disorder or condition. The term "treatment" as used herein, unless otherwise indicated, refers to the act of treating.

As used herein, the phrase "a method of treating" or its equivalent, when applied to, for example, cancer, refers to a procedure or course of action that is designed to reduce, eliminate, or inhibit the progression of the number of cancer cells in a patient; to induce differentiation of cancer cells or sensitize cancer cells to other therapeutic agents in a patient; and/or to alleviate the symptoms of a cancer. As used herein, the term "therapeutically effective amount" or "effective amount" means the amount of the subject compound, composition or combination that will elicit the biological or medical response of a cell, tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician. "Pharmaceutically acceptable salts", as used herein, refers to the relatively non-toxic, inorganic and organic acid addition salts, and base addition salts, of compounds of formula (I). These salts can be prepared in situ during the final isolation and purification of the compounds. Acid addition salts are a convenient form for use; and in practice, use of the salt form in essence amounts to use of the free base form. The acids which can be used to prepare the acid addition salts include preferably those which produce, when combined with the free base, pharmaceutically acceptable salts, that is, salts whose anions are non-toxic to the patient in pharmaceutical doses of the salts, so that the beneficial inhibitory effects inherent in the free base are not vitiated by side effects ascribable to the anions. Pharmaceutically acceptable salts within the scope of the invention include those derived from mineral acids and organic acids. See, for example S.M. Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci., 66, 1 -19 (1977).

Where the compound of the invention is substituted with an acidic moiety, base addition salts may be formed and are simply a convenient form for use; and in practice, use of the salt form in essence amounts to use of the free acid form. The bases which can be used to prepare the base addition salts include preferably those which produce, when combined with the free acid, pharmaceutically acceptable salts, that is, salts whose cations are non-toxic to the patient in pharmaceutical doses of the salts, so that the beneficial inhibitory effects inherent in the free base are not vitiated by side effects ascribable to the cations. Base addition salts can also be prepared by separately reacting the purified compound in its acid form with a suitable organic or inorganic base derived from alkali and alkaline earth metal salts and isolating the salt thus formed.

As well as being useful in themselves as active compounds, salts of compounds of the invention are useful for the purposes of purification of the compounds, for example by exploitation of the solubility differences between the salts and the parent compounds, side products and/or starting materials by techniques well known to those skilled in the art. It will be appreciated that the compounds of the invention may contain asymmetric centers. These asymmetric centers may independently be in either the R or S configuration. It will be apparent to those skilled in the art that certain compounds of formula (I) may also exhibit geometrical isomerism. It is to be understood that the present invention includes individual geometrical isomers and stereoisomers and mixtures thereof, including racemic mixtures, of compounds of formula (I) hereinabove. Such isomers can be separated from their mixtures, by the application or adaptation of known methods. Chiral chromatography techniques represent one means for separating isomers from mixtures thereof. Some compounds may be separated by chiral recrystallization techniques as an alternative means for separating isomers from mixtures thereof. Individual isomeric compounds can also be prepared by employing, where applicable, chiral precursors. Thus, in accordance with the practice of this invention there is provided a compound of the formula (I):

wherein:

W represents CR6R7, S, or O;

X represents a nitrogen atom and Y represents CRs; or X represents CR9 and Y represents a nitrogen atom;

Ri , R2, R3, R4, R5, Re, and R ₉ , are the same or different and independently selected from the group consisting of hydrogen, halogen, cyano, amino, monoalkylamino, dialkylamino, cyanoalkyl, carbamide, haloalkyl, alkoxyalkyl, alkylcarbonyl, an unsubstituted or substituted alkyl, an unsubstituted or substituted alkoxy, an unsubstituted or substituted thioalkyi, and an unsubstituted or substituted aminoalkyi; wherein the alkyl, alkoxy, thioalkyi or aminoalkyi substituents are one or more radicals independently selected from the group consisting of alkyl, alkoxy, thioalkyi, and aminoalkyi;

R6 is selected from the group consisting of halogen, cyano, cyanoalkyl, haloalkyl, benzyl, -NR _a R _b , alkylcarbonyl, alkoxycarbonyl, arylcarbonyl, heteroarylcarbonyl, an unsubstituted or substituted aryl, an unsubstituted or substituted heteroaryl, an unsubstituted or substituted alkyl, an unsubstituted or substituted alkoxy, an unsubstituted or substituted thioalkyi, and an unsubstituted or substituted aminoalkyi; wherein when said aryl, heteroaryl, alkyl, alkoxy, thioalkyi or aminoalkyi represented by R ₆ is substituted, the substituents are one or more radicals independently selected from the group consisting of alkyl, halogen, phenyl, alkoxy, thioalkyi, and aminoalkyi; or R ₃ and R ₆ , together with the carbon atoms to which they are attached, form a cycloalkyl group;

R ₇ is selected from the group consisting of hydrogen, alkyl, cyano, and halogen; and

R _a and R _b are the same or different and independently selected from the group consisting of hydrogen, alkyl, arylalkoxycarbonyl, and arylalkyl.

In one aspect of the invention, the compounds of formula (I) do not include compounds wherein W represents CR6R7, and Ri , R2, R3, R4, R5, R7, and R9 are hydrogen and Y is a nitrogen atom and R ₆ is ethyl or 1 ,1 -dimethylpropyl.

In another aspect of the invention, the compounds of formula (I) do not include the compounds selected from 8-ethyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole and 8- (1 ,1 -dimethylpropyl)-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole.

As already mentioned above, the compound of formula (I) may be present as a salt when such possibility exists. All forms of salts that can be envisaged are part of this invention. In another embodiment of this invention the compound of formula (I) has the following substituents:

Ri is selected from the group consisting of hydrogen, alkyl, alkoxyalkyl, and carbamide;

R ₂ is selected from the group consisting of hydrogen, alkyl, and cyano,

R ₃ is hydrogen;

R ₄ is selected from the group consisting of hydrogen and alkyl;

R ₅ is selected from the group consisting of hydrogen, alkyl, carbamide, and halogen;

R ₆ is selected from the group consisting of alkyl, haloalkyl benzyl, -NR _a R _b , alkylcarbonyl, alkoxycarbonyl, phenylcarbonyl, and an unsubstituted or substituted phenyl, wherein said phenyl substituents are one or more radicals selected from the group consisting of halogen and phenyl; or R ₃ and R ₆ , together with the carbon atoms to which they are attached, form a cycloalkyl group;

R ₇ is hydrogen, cyano, or alkyl;

Rs is hydrogen;

Rg is hydrogen; and

R _a and R _b independently represent benzyl;

or a salt thereof.

In an embodiment of this invention, the compound of this invention is represented by formula (la):

wherein W, R-i , R ₂ , R3, R4, R5, and Rs are as defined above for the compounds of formula (I); or a salt thereof.

In another embodiment, the compound of this invention can be represented by formula (lb):

wherein W, R-i , R ₂ , R3, R4, R5, and Rg are as defined above for the compounds of formula (I); or a salt thereof.

In another embodiment, the compound of this invention can be represented by formula (lc):

wherein W, X, Y, R-i , R ₂ , R3, R ₄ , and R5 are as defined above for the compounds of formula (I); or a salt thereof. In another embodiment, the compound of this invention can be represented by formula (Id):

wherein W, X, Y, R-i , R ₂ , R3, R ₄ , and R5 are as defined above for the compounds of formula (I); or a salt thereof.

In another embodiment, the compound of this invention can be represented by formula (le):

wherein X, Y, R^ R ₂ , R3, R ₄ , R5, R6 and R ₇ are as defined above for the compounds of formula (I); or a salt thereof.

As specific examples of compound of formula (I), the following compounds may be enumerated without any limitations:

8-tert-butyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole; 6-tert-butyl-6,7,8,9-tetrahydro-5H-beta-carboline;

8-ethyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

8,8-dipropyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

8-tert-butyl-4-methyl-6,7,8,9-tetrahydro-5H-pyrido[4, 3-b]indole;

8-tert-butyl-4-chloro-6,7,8,9-tetrahydro-5H-pyrido[4, 3-b]indole;

6-tert-butyl-9-methyl-6,7,8,9-tetrahydro-5H-beta-carbolin e;

8-tert-butyl-2,5-dimethyl-6,7,8,9-tetrahydro-5H-pyrido[4, 3-b]indol-2-ium;

(8R)-8-tert-butyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indo le;

(8S)-8-tert-butyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indo le;

8-trifluoromethyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indo le;

8-tert-butyl-5-methyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]ind ole;

8-tert-butyl-5-ethyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]i ndole;

6- tert-butyl-9-methyl-6,7,8,9-tetrahydro-5H-beta-carboline;

6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole-8-carboxylic acid ethyl ester;

8-methyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

8-isopropyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

8,8-dimethyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

dibenzyl-(6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indol-8-yl)- amine;

8-phenyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

8-(1 ,1 -dimethyl-propyl)-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

8-pentyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

8-tert-butyl-1 -methyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

2,3,4,4a,5,6,7,1 1 c-octahydro-1 H-benzo[e]pyrido[4,3-b]indole;

8-Methyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

9-Methyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

1 ,2,4,9-Tetrahydro-3-thia-6,9-diaza; and

7- Methyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole;

or a salt thereof. The compounds of this invention can be synthesized by any of the procedures known to one skilled in the art. Specifically, several of the starting materials used in the preparation of the compounds of this invention are known or are themselves commercially available. The compounds of this invention and several of the precursor compounds may also be prepared by methods used to prepare similar compounds as reported in the literature and as further described herein.

More specifically, the compounds disclosed herein can be synthesized according to the following procedures of Schemes 1 - 5, wherein W, X, Y, R-i , R ₂ , R3, R4, R5, R6, R ₇ , R ₈ , and R ₉ are as defined for formula (I) or are precursors thereto unless otherwise indicated.

Scheme 1 illustrates the synthesis of several of the compounds of formula (I) of this invention wherein R-i is hydrogen.

Scheme 1

Step 1 b

In Step 1 a, Scheme 1 , a ketone of formula (III) is deprotonated with NaHMDS or LiHMDS in a suitable solvent (for example, Et ₂ O or DCM). Trifluoromethanesulfonic anhydride (Tf ₂ O) is then added into the mixture. This reaction is generally carried out at a suitable temperature, for example between about -20°C to about -15°C. As an alternative to Step 1 a, in Step 1 b trifluoromethanesulfonic anhydride is added to a solution of the compound of formula (III) and 2,6-di-tert-butyl-4-methylpyridine. The reaction is carried out in a suitable solvent, for example DCM or THF, and is carried out at suitable temperatures, for example at about 0°C to about 5°C or up to ambient temperature.

In Step 2, Scheme 1 , an aminochloropyridine and a triflate are heated with a Pd catalyst (for example, Pd(PPh ₃ ) , Pd(dppf)CI ₂ CH ₂ CI ₂ , or Pd ₂ (dba) ₃ CHCI ₃ ), a ligand (for example, 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl) and a base (for example, Cs ₂ CO ₃ , NaOt-Bu, or K ₂ CO ₃ ), in an organic solvent (for example, 1 ,4- dioxane, THF, DME, DMF, toluene, or t-BuOH), at elevated temperatures, (about 80°C to about 100°C, for example) in an inert atmosphere.

Scheme 2 illustrates another approach for the preparation of compounds of this invention.

Scheme 2

In Scheme 2, step 1 , the 4-amino-3-iodopyridine is reacted with a ketone of formula (IV), in the presence of Si(OEt) and PPTS. The reaction is typically carried out in a suitable solvent, such as DMF or pyridine, and at elevated temperature (about 160°C to about 180°C, for example), for example in the presence of microwaves.

In Step 2, the imine of formula (V) is cyclized in the presence of catalytic Pd(PPh ₃ ) and Cy ₂ NMe. The reaction is carried out in a suitable solvent, such as DMF or pyridine and at elevated temperature (about 160°C to about 180°C, for example), in the presence of microwaves for example.

Alternatively, step 2 is performed without isolating the imine of formula (V). Scheme 3 provides another approach for the preparation of the compounds of formula (I) wherein Ri is hydrogen.

Scheme 3

In Scheme 3, step 1 , pyridinylhydrazine is reacted with a ketone of formula (IV). The reaction is carried out in a suitable solvent, such as an alcohol, for example, ethanol. The reaction is generally carried out at elevated temperature, for example between about 80°C to about 100°C.

For step 2, the compound of formula (VI) is cyclized in a suitable solvent, such as diethylene glycol, and at elevated temperatures, for example between about 200°C and 250°C. The reaction is optionally performed in the present of a catalytic amount of a bronsted acid or lewis acid, for example ZnC^. Scheme 4 further illustrates another variation for the preparation of compounds of formula (I) of this invention. heme 4

The reaction of Scheme 4 entails reacting a 4-amino-3-iodopyridine with the ketone of formula (VII). This reaction is performed in the presence of a base, for example CS2CO3, K2CO3, and DABCO, and a Pd catalyst for example Pd(OAc)2, and is carried out in a suitable solvent, such as DMF or DMA. The reaction is performed at elevated temperatures, for example at about 140°C.

Scheme 5 illustrates an approach for the preparation of compounds of formula (I) of this invention wherein Ri is, for example, an alkyl or alkylcarbonyl group.

Scheme 5

In Scheme 5, the compound of formula (la) is exposed to NaH in a suitable solvent, such as DMF or THF. This step is typically carried out at a temperature between about 0°C and room temperature. The reaction mixture is next exposed to, for example, an alkyl halide or alkylcabonyl halide, generally at temperatures between about 0°C and room temperature.

The present invention provide methods for treating Eg5-mediated diseases including cellular proliferative disorders, such as cancer, in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound of the invention.

Cellular proliferative disorders include, but are not limited to, cancer, tumor, smooth muscle cell proliferation, hyperplasia, angiogenesis, synovitis, restenosis, cardiac hyperplasia, immune disorder and inflammation.

Another aspect of the invention is a method for treating cancer in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound of the invention.

In particular, the invention provides methods for treating various types of cancers. More specifically, the compounds of formula (I) can be useful in the treatment of a variety of cancers, including, but not limited to, malignancies derived from hematopoietic cells and solid tumors.

In one aspect of the invention, the cancer to be treated is selected from breast cancer, bone cancer, lung cancer, oral cancer, cancer of the esophagus, cancer of the small intestine, colon cancer, rectal cancer, cancer of the anal region, liver cancer, stomach cancer, cancer of the bile ducts or bile vesicle, pancreatic cancer, cancer of the urinary pathways (for example, cancer of the kidneys, ureter, or bladder), cancer of female genital organs, (for example, cancer of the uterus, cervix or ovaries), cancers of male genital organs (for example, cancer of the penis, prostate cancer, cancer of the seminal vesicles or of the testicles), cancer of the endocrine system (for example, cancer of the thyroid, pituitary or adrenal glands), skin cancer, cancer of the head and neck, tumors of the central and peripheral nervous system (for example, neuroblastomas, schwannomas, and meningiomas), hematopoietic malignancies including leukemia, such as acute lymphocytic leukemia (ALL), acute myeloid (myelogenous) leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), hairy cell leukemia (HCL), chloromas, prolymphocytic leukemia, and T-cell leukemia; and lymphoma, such as non-Hodgkin lymphoma, B- cell lymphoma, T-cell lymphoma, NK cell lymphoma, Hodgkin lymphoma, and Burkitt's lymphoma and mantle cell lymphoma; multiple myeloma, myelodysplasia syndromes (MDS), and Waldenstrom's Macroglobulinemia.

Another aspect of the invention is a method for treating cancer in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound of the invention, wherein the cancer is selected from the group consisting of breast cancer, bone cancer, lung cancer, oral cancer, cancer of the esophagus, cancer of the small intestine, colon cancer, rectal cancer, cancer of the anal region, liver cancer, stomach cancer, cancer of the bile ducts or bile vesicle, pancreatic cancer, cancer of the urinary pathways, cancer of female genital organs, cancer of male genital organs, prostate cancer, cancer of the endocrine system, skin cancer, cancer of the head and neck, tumors of the central and peripheral nervous system, leukemia, acute lymphocytic leukemia, acute myeloid (myelogenous) leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, chloroma, prolymphocytic leukemia, T-cell leukemia, non-Hodgkin lymphoma, B-cell lymphoma, T-cell lymphoma, NK cell lymphoma, Hodgkin lymphoma, Burkitt's lymphoma, multiple myeloma, myelodysplasia syndromes (MDS), and Waldenstrom's Macroglobulinemia.

In a particular aspect, the present invention relates to the use of compounds of the invention for the preparation of a medicinal product intended for treating cellular proliferation disorders, such as cancers.

In one aspect, the present invention relates to a medicament, characterized in that it comprises a compound of the invention.

A particular aspect of the invention is the use of a compound of the invention for the treatment of cellular proliferation disorders, such as cancer. Another aspect of the invention is a method for inhibiting the activity of Eg5, which comprises contacting Eg5 with a compound of the invention in an amount effective to inhibit Eg5. In one embodiment, the contacting occurs in vivo. In another embodiment, the contacting occurs in a biological sample.

Inhibition of Eg5 activity in a biological sample is useful for various purposes, including, but not limited to, storage of biological specimens, study of gene regulatory pathways, characterization of signal transduction cascades, and identification of potential downstream drugable targets.

The compounds of the invention are useful for treating cancer in patients, including humans. When administered to a patient, the compounds can be used alone, or as a pharmaceutical composition. The compounds of the invention can also be administered in combination with one or more additional anticancer active ingredients.

The present invention also includes within its scope pharmaceutical compositions comprising at least one of the compounds of the invention in admixture with a pharmaceutically acceptable carrier. In practice, the compounds of the invention may be administered in pharmaceutically acceptable dosage forms to humans and other animals by topical or systemic administration, including oral, inhalational, rectal, nasal, buccal, sublingual, vaginal, colonic, parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural), intracisternal and intraperitoneal. It will be appreciated that the preferred route may vary with for example the condition of the recipient.

"Pharmaceutically acceptable dosage forms" refers to dosage forms of the compound of the invention, and includes, for example, tablets, dragees, powders, elixirs, syrups, liquid preparations, including suspensions, sprays, inhalants, lozenges, emulsions, solutions, granules, capsules and suppositories, as well as liquid preparations for injections, including liposome preparations. Techniques and formulations generally may be found in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, latest edition. A particular aspect of the invention provides for a compound of the invention to be administered in the form of a pharmaceutical composition. Pharmaceutical compositions, according to the present invention, comprise one or more compounds of the invention and pharmaceutically acceptable carriers.

Pharmaceutically acceptable carriers include one or more component selected from the group comprising pharmaceutically acceptable carriers, diluents, coatings, adjuvants, excipients, or vehicles, such as preserving agents, fillers, disintegrating agents, wetting agents, emulsifying agents, emulsion stabilizing agents, suspending agents, isotonic agents, sweetening agents, flavoring agents, perfuming agents, coloring agents, antibacterial agents, antifungal agents, other therapeutic agents, lubricating agents, adsorption delaying or promoting agents, and dispensing agents, depending on the nature of the mode of administration and dosage forms.

Exemplary suspending agents include ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances.

Exemplary antibacterial and antifungal agents for the prevention of the action of microorganisms include parabens, chlorobutanol, phenol, sorbic acid, and the like.

Exemplary isotonic agents include sugars, sodium chloride, and the like.

Exemplary adsorption delaying agents to prolong absorption include aluminum monostearate and gelatin.

Exemplary adsorption promoting agents to enhance absorption include dimethyl sulfoxide and related analogs.

Exemplary diluents, solvents, vehicles, solubilizing agents, emulsifiers and emulsion stabilizers, include water, chloroform, sucrose, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, tetrahydrofurfuryl alcohol, benzyl benzoate, polyols, propylene glycol, 1 ,3-butylene glycol, glycerol, polyethylene glycols, dimethylformamide, TWEEN™ 60, SPAN™ 60, cetostearyl alcohol, myristyl alcohol, glyceryl mono-stearate and sodium lauryl sulfate, fatty acid esters of sorbitan, vegetable oils (such as cottonseed oil, groundnut oil, com germ oil, olive oil, castor oil and sesame oil) and injectable organic esters such as ethyl oleate, and the like, or suitable mixtures of these substances.

Exemplary excipients include lactose, milk sugar, sodium citrate, calcium carbonate and dicalcium phosphate.

Exemplary disintegrating agents include starch, alginic acids and certain complex silicates. Exemplary lubricants include magnesium stearate, sodium lauryl sulfate, talc, as well as high molecular weight polyethylene glycols.

The choice of pharmaceutically acceptable carrier is generally determined in accordance with the chemical properties of the active compound such as solubility, the particular mode of administration and the provisions to be observed in pharmaceutical practice.

Pharmaceutical compositions of the present invention suitable for oral administration may be presented as discrete units such as a solid dosage form, such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient, or as a powder or granules; as a liquid dosage form such as a solution or a 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 active ingredient may also be presented as a bolus, electuary or paste.

"Solid dosage form" means the dosage form of the compound of the invention is in solid form, for example capsules, tablets, pills, powders, dragees or granules. In such solid dosage forms, the compound of the invention is admixed with at least one inert customary excipient (or carrier) such as sodium citrate or dicalcium phosphate or (a) fillers or extenders, as for example, starches, lactose, sucrose, glucose, mannitol and silicic acid, (b) binders, as for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia, (c) humectants, as for example, glycerol, (d) disintegrating agents, as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates and Na2CO3, (e) solution retarders, as for example paraffin, (f) absorption accelerators, as for example, quaternary ammonium compounds, (g) wetting agents, as for example, cetyl alcohol and glycerol monostearate, (h) adsorbents, as for example, kaolin and bentonite, (i) lubricants, as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, (j) opacifying agents, (k) buffering agents, and agents which release the compound(s) of the invention in a certain part of the intestinal tract in a delayed manner. A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tables may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface active or dispersing agent. Excipients such as lactose, sodium citrate, calcium carbonate, dicalcium phosphate and disintegrating agents such as starch, alginic acids and certain complex silicates combined with lubricants such as magnesium stearate, sodium lauryl sulfate and talc may be used. A mixture of the powdered compounds moistened with an inert liquid diluent may be molded in a suitable machine to make molded tablets. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein.

Solid compositions may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols, and the like.

If desired, and for more effective distribution, the compounds can be microencapsulated in, or attached to, slow release or targeted delivery systems such as biocompatible, biodegradable polymer matrices (e.g., poly(d,l-lactide co- glycolide)), liposomes, and microspheres and subcutaneously or intramuscularly injected by a technique called subcutaneous or intramuscular depot to provide continuous slow release of the compound(s) for a period of 2 weeks or longer. The compounds may be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water, or some other sterile injectable medium immediately before use. "Liquid dosage form" means the dose of the active compound to be administered to the patient is in liquid form, for, example, pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art, such as solvents, solubilizing agents and emulsifiers.

When aqueous suspensions are used they can contain emulsifying agents or agents which facilitate suspension.

Pharmaceutical compositions suitable for topical administration mean formulations that are in a form suitable to be administered topically to a patient. The formulation may be presented as a topical ointment, salves, powders, sprays and inhalants, gels (water or alcohol based), creams, as is generally known in the art, or incorporated into a matrix base for application in a patch, which would allow a controlled release of compound through the transdermal barrier. When formulated in an ointment, the active ingredients may be employed with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredients may be formulated in a cream with an oil-in-water cream base. Formulations suitable for topical administration in the eye include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient. Formulations suitable for topical administration in the mouth include lozenges comprising the active ingredient in a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

The oily phase of the emulsion pharmaceutical composition may be constituted from known ingredients in a known manner. While the phase may comprise merely an emulsifier (otherwise known as an emulgent), it desirably comprises a mixture of at least one emulsifier with a fat or an oil or with both a fat and an oil. In a particular embodiment, a hydrophilic emulsifier is included together with a lipophilic emulsifier that acts as a stabilizer. Together, the emulsifier(s) with or without stabilizer(s) make up the emulsifying wax, and together with the oil and fat make up the emulsifying ointment base which forms the oily dispersed phase of the cream formulations.

If desired, the aqueous phase of the cream base may include, for example, at least 30% w/w of a polyhydric alcohol, i.e. an alcohol having two or more hydroxy groups such as propylene glycol, butane 1 ,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol (including PEG 400) and mixtures thereof. The topical formulations may desirably include a compound that enhances absorption or penetration of the active ingredient through the skin or other affected areas. The choice of suitable oils or fats for a composition is based on achieving the desired properties. Thus a cream should preferably be a non-greasy, non-staining and washable product with suitable consistency to avoid leakage from tubes or other containers. Straight or branched chain, mono- or dibasic alkyl esters such as di- isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a blend of branched chain esters known as Crodamol CAP may be used. These may be used alone or in combination depending on the properties required. Alternatively, high melting point lipids such as white soft paraffin and/or liquid paraffin or other mineral oils can be used. Pharmaceutical compositions suitable for rectal or vaginal administrations means formulations that are in a form suitable to be administered rectally or vaginally to a patient and containing at least one compound of the invention. Suppositories are a particular form for such formulations that can be prepared by mixing the compounds of the invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at ordinary temperatures but liquid at body temperature and therefore, melt in the rectum or vaginal cavity and release the active component.

Pharmaceutical composition administered by injection may be by transmuscular, intravenous, intraperitoneal, and/or subcutaneous injection. The compositions of the present invention are formulated in liquid solutions, in particular in physiologically compatible buffers such as Hank's solution or Ringer's solution. In addition, the compositions may be formulated in solid form and redissolved or suspended immediately prior to use. Lyophilized forms are also included. The formulations are sterile and include emulsions, suspensions, aqueous and non-aqueous injection solutions, which may contain suspending agents and thickening agents and antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic, and have a suitably adjusted pH, with the blood of the intended recipient.

Pharmaceutical composition of the present invention suitable for nasal or inhalational administration means compositions that are in a form suitable to be administered nasally or by inhalation to a patient. The composition may contain a carrier, in a powder form, having a particle size for example in the range 1 to 500 microns (including particle sizes in a range between 20 and 500 microns in increments of 5 microns such as 30 microns, 35 microns, etc.). Suitable compositions wherein the carrier is a liquid, for administration as for example a nasal spray or as nasal drops, include aqueous or oily solutions of the active ingredient. Compositions suitable for aerosol administration may be prepared according to conventional methods and may be delivered with other therapeutic agents. Metered dose inhalers are useful for administering compositions according to the invention for an inhalational therapy.

Actual dosage levels of active ingredient(s) in the compositions of the invention may be varied so as to obtain an amount of active ingredient(s) that is (are) effective to obtain a desired therapeutic response for a particular composition and method of administration for a patient. A selected dosage level for any particular patient therefore depends upon a variety of factors including the desired therapeutic effect, on the route of administration, on the desired duration of treatment, the etiology and severity of the disease, the patient's condition, weight, sex, diet and age, the type and potency of each active ingredient, rates of absorption, metabolism and/or excretion and other factors.

Total daily dose of the compounds of the invention administered to a patient in single or divided doses may be in amounts, for example, of from about 0.001 to about 100 mg/kg body weight daily and preferably 0.01 to 10 mg/kg/day. For example, in an adult, the doses are generally from about 0.01 to about 100, preferably about 0.01 to about 10, mg/kg body weight per day by inhalation, from about 0.01 to about 100, preferably 0.1 to 70, more especially 0.5 to 10, mg/kg body weight per day by oral administration, and from about 0.01 to about 50, preferably 0.01 to 10, mg/kg body weight per day by intravenous administration. The percentage of active ingredient in a composition may be varied, though it should constitute a proportion such that a suitable dosage shall be obtained. Dosage unit compositions may contain such amounts of such submultiples thereof as may be used to make up the daily dose. Several unit dosage forms may be administered at about the same time. A dosage may be administered as frequently as necessary in order to obtain the desired therapeutic effect. Some patients may respond rapidly to a higher or lower dose and may find much weaker maintenance doses adequate. For other patients, it may be necessary to have long-term treatments at the rate of 1 to 4 doses per day, in accordance with the physiological requirements of each particular patient. For other patients, it will be necessary to prescribe not more than one or two doses per day. The formulations can be prepared in unit dosage form by any of the methods well known in the art of pharmacy. Such methods include the step of bringing into association the active ingredient with the carrier that constitutes one or more accessory ingredients. In general the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials with elastomeric stoppers, and may be stored in a freeze- dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

It should be understood that the compounds of the invention can be used alone or in combination with an additional agent, e.g., a therapeutic agent, said additional agent being selected by the skilled artisan for its intended purpose. For example, the additional agent can be a therapeutic agent art-recognized as being useful to treat the disease or condition being treated by the compound of the present invention. The additional agent also can be an agent that imparts a beneficial attribute to the therapeutic composition.

Other embodiments of the invention are the methods of preparing the compounds of the invention as herein described.

The compounds of this invention can be synthesized by any of the procedures known to one skilled in the art. Specifically, several of the starting materials used in the preparation of the compounds of this invention are known or are themselves commercially available. The compounds of this invention and several of the precursor compounds may also be prepared by methods used to prepare similar compounds as reported in the literature and as further described herein. For instance, see R. C. Larock, "Comprehensive Organic Transformations," VCH publishers, 1989.

It is also well known that in various organic reactions it may be necessary to protect reactive functional groups, such as for example, amino groups, to avoid their unwanted participation in the reactions. Conventional protecting groups may be used in accordance with standard practice and known to one skilled in the art, for example, see T. W. Greene and P. G. M. Wuts in "Protective Groups in Organic Chemistry" John Wiley and Sons, Inc., 1991 . For example, suitable amine protecting groups include without any limitation sulfonyl (e.g., tosyl), acyl (e.g., benzyloxycarbonyl or t- butoxycarbonyl) and arylalkyl (e.g., benzyl), which may be removed subsequently by hydrolysis or hydrogenation as appropriate. Other suitable amine protecting groups include trifluoroacetyl [-C(=O)CF ₃ ] which may be removed by base catalyzed hydrolysis, or a solid phase resin bound benzyl group, such as a Merrifield resin bound 2,6-dimethoxybenzyl group (Ellman linker) or a 2,6-dimethoxy-4-[2- (polystyrylmethoxy)ethoxy]benzyl, which may be removed by acid catalyzed hydrolysis, for example with TFA.

According to a further feature of the invention, acid addition salts of the compounds of the invention may be prepared by reaction of the free base with the appropriate acid, by the application or adaptation of known methods. For example, the acid addition salts of the compounds of the invention may be prepared either by dissolving the free base in water or aqueous alcohol solution or other suitable solvents containing the appropriate acid and isolating the salt by evaporating the solution, or by reacting the free base and acid in an organic solvent, in which case the salt separates directly or can be obtained by concentration of the solution.

Compounds of the invention can be regenerated from their acid addition salts by the application or adaptation of known methods. For example, parent compounds of the invention can be regenerated from their acid addition salts by treatment with an alkali, e.g., aqueous sodium bicarbonate solution.

Compounds of the invention can be regenerated from their base addition salts by the application or adaptation of known methods. For example, parent compounds of the invention can be regenerated from their base addition salts by treatment with an acid, e.g., hydrochloric acid.

The compounds of the invention, their methods of preparation and their biological activity will appear more clearly from the examination of the following examples that are presented as an illustration only and are not to be considered as limiting the invention in its scope. Experimental Procedures

The starting materials and intermediates may be prepared by the methods described in the present application or are available commercially or are described in the literature, or else may be prepared by methods which are described therein or which are known to a person skilled in the art. By appropriate selection of suitable starting materials, compounds of the invention may be prepared according to the procedures described in the foregoing examples.

The examples which follow describe the preparation of certain compounds in accordance with the invention. These examples are not limitative, and merely illustrate the present invention. The numbers of the compounds exemplified match those given in the tables hereinafter, which illustrates the chemical structures and physical properties of some compounds according to the invention. Compounds of the invention are identified, for example, by the following analytical methods.

High Pressure Liquid Chromatography - Mass Spectrometry (LCMS) experiments to determine retention times (RT) and associated mass ions are performed using one of the following analytical methods.

Method A: YMC Jsphere H80, 33 ^* 2, 4u, H ₂ O + 0.05% TFA: acetonitrile + 0.05% TFA 98:2 (1 min) to 5:95 (5.0min) to 5:95 (6.25min). Ionization method: ES+ Method B: WatersXBridge C18, 4.6 ^* 50, 2.5μ, H ₂ O + 0.05% TFA: acetonitrile + 0.05% TFA 95:5 (Omin) to 95:5 (0.3min) to 5:95 (3.5 min) to 5:95 (4min). Ionization method: ES+

Method C: Waters XBridge C18 4.6 ^* 50 mm; 2.5μ, H ₂ O+0.1 % TFA:acetonitrile + 0.08% TFA 97:3 (Omin) to 40:60 (3.5 min) to 2:98 (4min) to 2:98 (5min) to 97:3 (5.2min) to 97:3 (6.5min). Ionization method: ES+. Method D: SYNERGI 2U HYDRO-RP 20X2.0MM, 0.1 % formic acid in Water/Acetonitrile, 5 to 40% acetonitrile in 2 min., to 95% acetonitrile at 5 min. Ionization method: ESI+, MIC ROM ASS LCT-LCMS, scan m/z 100-1200.

Method E: Waters Acquity-SQD UPLC-MS System, 2.1 x 50mm, 1 .7micron, BEH-C18 Acquity column, 55°C, water + 0.05 % formic acid/acetonitrile + 0.05% Formic Acid, 0.8 mL/min, run 1 .5 min. Ionization method: SQD Single Quadrupole Detector, Tunable Visible/Ultra-violet (TUV) Dual Wavelength Detector, ESI +/- (Electrospray Ionization dual positive/negative), APCI +/- (Atmospheric Pressure Chemical Ionization dual positive/negative).

Method F: :SYNERGI 2U HYDRO-RP 20x2.0MM, A=H2O,B=ACN,+0.1 %Formic Acid ;grad 3-98%B/7min. IONIZATION METHOD=ESI+;UV WAVELENGTH=220.

All liquid chromatography coupled mass spectroscopy spectra (LCMS) express a retention time (RT) in minutes (min).

¹ H NMR spectra were measured on a Varian Mercury (300 MHz) spectrometer using tetramethylsilane and the deuterated solvent respectively as internal standards.

Preparative separations were carried out using a Gemini 10μ chromatography system, using a C18 reverse phase column, 1 1 OA, 150x30 (Typical solvents were varying concentrations of acetonitrile in water with 0.1 % TFA, e.g. a 5% to 90% gradient of acetonitrile/0.01 % aqueous TFA over 40 min at 20 mL/min).

Example 1

8-tert-Butyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole Trifluoromethanesulfonic acid 4-tert-butyl-cyclohex-1 -enyl ester

To an ice-cooled solution of 4-tert-butylcyclohexanone (17.0 g, 1 10.5 mmol) and 2,6- di-tert-butyl-4-methylpyridine (26.63 g, 129.7 mmol) in DCM (575 ml), was added dropwise trifluoromethanesulfonic anhydride (37.6 g, 133.3 mmol). After completion of the addition, the mixture was stirred and allowed to warm up to room temperature overnight. The resulting white precipitate was filtered off and washed with a small amount of heptane. The solution was washed with 1 N HCI (aq), 1 N NaOH (aq) and brine, respectively. The organic phase was concentrated and subjected to chromatography (silica gel, eluting with 100% heptane), to afford trifluoromethanesulfonic acid 4-tert-butyl-cyclohex-1 -enyl ester as a clear oil (16.23 g, 51 %).

Step 2: 8-tert-But l-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole

4-Amino-3-chloropyridine (8.0 g, 62 mmol) and K2CO3 (19.56 g, 142 mmol) were added into a round bottom pressure vessel, followed by addition of t-BuOH (100 ml). The mixture was stirred at room temperature and purged with N ₂ for 10min. Pd ₂ (dba) ₃ CHCI ₃ (2.929 g, 2.83 mmol) and 2-di-tert-butylphosphino-2',4',6'- triisopropylbiphenyl (4.807 g, 1 1 .32 mmol) were added. The mixture was stirred at room temperature for another 10 min before trifluoromethanesulfonic acid 4-tert-butyl- cyclohex-1 -enyl ester (16.209 g, 56.6 mmol) was added. The pressure vessel was sealed under N ₂ and heated at 80~85°C for 43 hours. The reaction mixture was cooled, diluted with EtOAc and filtered through Celite. The filtrate was concentrated and triturated with DCM/EtOAc. The resulting solid was filtered and washed with DCM and H ₂ O, respectively. The beige solid was dried under vacuum and afforded 3.07g of the title compound. The filtrate was concentrated, and the residue was subjected to chromatography (silica gel treated with Et ₃ N, eluting with MeOH in DCM). Another 0.50g of 8-tert-butyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole was obtained as a beige solid (total yield: 28%). Example 2

6-tert-But l-6,7,8,9-tetrahydro-5H-beta-carboline

The title compound was prepared from 3-amino-4-chloropyridine (7.41 g, 57.6 mmol) and trifluoromethanesulfonic acid 4-tert-butylcyclohex-1 -enyl ester (15.0 g, 52.4 mmol) following the procedure of Example 1 , Step 2, to afford the desired product as a beige solid (1 .02 g, 9%).

Example 3

8-Eth l-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole

Si(OEt) (284 mg, 1 .36 mmol) was added to a solution of 4-amino-3-iodopyridine (300 mg, 1 .36 mmol), 4-ethylcyclohexanone (344 mg, 2.72 mmol) and PPTS (68 mg, 0.27 mmol) in pyridine (3 ml). The mixture was flushed with N ₂ and heated at 160°C under microwave for 45 min. Pd(PPh ₃ ) (78 mg, 0.068 mmol) and dicyclohexylmethylamine (319 mg, 1 .63 mmol) were added to the reaction mixture. The mixture was flushed with N ₂ , then heated at 170°C under microwave for 2 hours. The reaction mixture was cooled and partitioned between H ₂ O and EtOAc. The organic phase was washed with brine, dried over Na ₂ SO ₄ and concentrated. The residue was subjected to chromatography (silica gel treated with Et ₃ N in DCM, eluting with MeOH in DCM) to afford 220 mg of impure product, which was purified again with HPLC (Ci _8, eluting with 10-100% CH ₃ CN in H ₂ O with 0.1 % TFA). 142 mg of gel was obtained from pure fractions. The gel was treated with MeOH and concentrated to give the title compound as a white crystalline solid (142 mg, TFA salt, 33%, hygroscopic).

Example 4

8,8-Dipropyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole

Step 1 : N-(4,4-Dipropylcyclohexylidene)-N'-pyridin-4-ylhydrazine

A solution of pyridin-4-ylhydrazine (1 .0 g, 9.2 mmol) and 4,4-dipropylcyclohexanone (1 .67 g, 9.2 mmol) in EtOH (10 ml) was heated at 90°C in a sealed tube under N ₂ for 4.5 hours. The mixture was cooled and concentrated. The crude intermediate N- (4,4-dipropyl-cyclohexylidene)-N'-pyridin-4-ylhydrazine was obtained as 2.55 g of an off-white solid and was used for the next step without purification.

Step 2: 8,8-Dipropyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole

N-(4,4-Dipropylcyclohexylidene)-N'-pyridin-4-ylhydrazine (0.95 g, 3.47 mmol) was heated in diethylene glycol (12 ml) at 250°C under microwave for 5 hours. The mixture was cooled and partitioned between H ₂ O/DCM/EtOAc. The organic phase was concentrated to afford 0.824 g of a red oil which was subjected to chromatography (silica gel neutralized with Et ₃ N in DCM, eluting with 10% MeOH in DCM). The title compound was obtained as an off-white solid (175 mg, 7% over 2 steps). Example 5

Butyl-4-methyl-6,7,8,9-tetrahydro-5H-pyrido[4, 3-b]indole

A suspension of 4-amino-3-iodo-5-methylpyridine (500 mg, 1 .07 mmol), 4-tert- butylcyclohexanone (497 mg, 3.22 mmol), DABCO (361 mg, 3.2 2mmol) in DMA (3 ml) in a vial was purged with N ₂ five times before Pd(OAc)2 (12 mg, 0.053 mmol) was added. The vial was sealed under N ₂ and heated at 140°C for 2.5 days. The reaction mixture was cooled and partitioned between H ₂ O/EtOAc (50ml/50ml). The mixture was filtered through Celite, and the aqueous phase was extracted with EtOAc (75ml x 7). The combined organic phases were dried over MgSO ₄ and concentrated. The residue was dissolved in methanol and purified with preparative HPLC (ds, eluting with CH ₃ CN in H ₂ O with 0.1 % TFA). The title compound was obtained as a TFA salt (3mg, <1 %).

Example 6

8-tert-Butyl-4-chloro-6,7,8,9-tetrahydro-5H-pyrido[4, 3-b]indole

A suspension of 3-chloro-4-amino-5-iodopyridine (2.0 g, 7.87 mmol), 4-tert- butylcyclohexanone (3.64 g, 23.6 mmol), potassium tert-butoxide (1 .148 g, 10.23 mmol), MgSO ₄ (0.474 g, 3.94 mmol) and acetic acid (0.708 g, 1 1 .8 mmol) in DMA (25 ml) was bubbled with N ₂ before Pd(OAc) ₂ (88 mg, 0.393 mmol) was added. The mixture was heated overnight at 140°C. The reaction mixture was partitioned with H ₂ O/EtOAc (100 ml/100 ml). The aqueous phase was extracted with EtOAc (100 ml x 5). The combined organic phases were washed with brine, dried over Na ₂ SO ₄ and concentrated. The residue was subjected to preparative HPLC (ds, eluting with CH ₃ CN in H ₂ O with 0.1 % TFA). The title compound was obtained as a TFA salt (38mg, 1 %).

Example 7

6-tert-Butyl-9-methyl-6,7,8,9-tetrahydro-5H-beta-carboline

To a suspension of NaH (41 mg of a 60% dispersion in mineral oil, 1 .02 mmol) in THF (1 ml), was added dropwise a solution of 6-tert-butyl-6,7,8,9-tetrahydro-5H-beta- carboline (150 mg, 0.66 mmol) in THF (3 ml). The mixture was stirred at room temperature for 1 hour. Mel solution (2.0M in MTBE, 0.46 ml, 0.924 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours. The mixture was quenched with NH CI (aq, saturated). The aqueous phase was extracted with Et ₂ O. The combined organic phases were washed with brine, dried over Na ₂ SO ₄ and concentrated. The residue was subjected to chromatography (silica gel, eluting with MeOH in DCM) to afford the title compound as an off-white solid (44 mg, 28%). Example 8

8-tert-But l-2,5-dimethyl-6,7,8,9-tetrahydro-5H-pyrido[4, 3-b]indol-2-ium

A mixture of 8-tert-butyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole (50 mg, 0.219 mmol), tetrabutylammonium hydrosulfate (3 mg, 0.007 mmol) and NaOH (10 mg, 0.263 mmol) in CH ₃ CN (1 ml) was stirred in a vial under N ₂ at room temperature for 20 min. Mel (37 mg, 0.263 mmol) was added, and the vial was sealed and heated at 40°C for 2h. The solvent was then removed under vacuum, and the residue was partitioned with H ₂ O/DCM. The aqueous phase was extracted with DCM (40 ml x 2). The combined organic phases were washed with brine, dried over MgSO ₄ , and concentrated. The residue was treated with ether and concentrated to afford the title compound as a hygroscopic solid (84mg, 100%).

Example 9

(8R)-8-tert-Butyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole Step 1 : (2S, 8'R)-1 -(8'-tert-Butyl-6',7',8',9'-tetrahydropyrido[4',3'-b]indol-5 '-yl)-3,3,3- trifluoro-2-methoxy-2-phenyl-propan-1 -one and (2S, 8'S)-1 -(8'-tert-butyl-6',7',8',9'- tetrahydropyrido[4',3'-b]indol-5'-yl)-3,3,3-trifluoro-2-meth oxy-2-phenyl-propan-1 -one

To a mixture of 8-tert-butyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole (356 mg, 1 .56 mmol) in DCM (20 ml) was added dimethylaminopyridine (19 mg, 0.156 mmol) at room temperature. The mixture was cooled to 0°C and triethylamine (0.33 ml, 2.34 mmol) was added, followed by addition of 3,3,3-trifluoro-2-methoxy-2-phenylpropionyl chloride (433 mg, 1 .72 mmol). The mixture was stirred at 0°C for 10 minutes and then allowed to warm up and stir at room temperature overnight. The reaction was quenched with saturated NH CI solution (aq.) and extracted with EtOAc. The organic phase was dried over MgSO ₄ and concentrated to afford 290 mg of residue containing the desired compound. Separation by chiral chromatography afforded the two diastereomeric intermediates, (2S, 8'R)-1 -(8'-tert-Butyl-6',7',8',9'- tetrahydropyrido[4',3'-b]indol-5'-yl)-3,3,3-trifluoro-2-meth oxy-2-phenyl-propan-1 -one and (2S, 8'S)-1 -(8'-tert-butyl-6',7',8',9'-tetrahydropyrido[4',3'-b]indol-5 '-yl)-3,3,3- trifluoro-2-methoxy-2-phenyl-propan-1 -one as white foams (42% overall yield, stereochemistry at C-8' nominally assigned).

Step 2: (8R)-8-tert-Butyl-6,7,8,9-tetrahydro-5H-pyrido[4,3-b]indole

A suspension of (2S, 8'R)-1 -(8'-tert-Butyl-6',7',8',9'-tetrahydropyhdo[4',3'-b]indol-5' - yl)-3,3,3-thfluoro-2-methoxy-2-phenyl-propan-1 -one (15 mg, 0.034 mmol) and LiOH (30 mg, 0.7 mmol) in THF (0.5ml) was stirred at room temperature for 2 hours. The reaction mixture was then quenched with water. The aqueous phase was extracted with EtOAc. The combined organic phases were washed with brine, dried over MgSO ₄ and concentrated. The residue was subjected to chromatography (silica gel, eluting with 5% MeOH in DCM), to afford compound the title compound as a colorless solid (8 mg, quantitative, stereochemistry at C-8 nominally assigned)

Example 10

(8S)-8-tert-Butyl-6,7,8,9-tetrah dro-5H-pyrido[4,3-b]indole

The title compound was prepared from (2S, 8'S)-1 -(8'-tert-butyl-6',7',8',9'- tetrahydropyrido[4',3'-b]indol-5'-yl)-3,3,3-trifluoro-2-meth oxy-2-phenyl-propan-1 -one (70mg, 0.16mmol), following the procedure of Example 9, to afford a pale yellow foam (35mg, quantitative, stereochemistry at C-8 nominally assigned). Table 1 below illustrates several intermediate compounds of the present invention and their physical properties.

Intermediate

Compound Structure Name Data

Number

0 3-(4-Chloro-pyridin- MS (method D): m/z =

3-ylamino)- 222.99 (ES+) cyclohex-2-enone ¹ H NMR (300 MHz,

DMSO-d ₆ ) δ 8.82 (s,

9a 1 H), 8.52 (s, 1 H), 8.43

(d, 1 H), 7.68 (d, 1 H), 4.62 (s, 1 H), 2.55 (m, 2H), 2.15 (t, 2H), 1 .91 (m, 2H)

Table II: Table II illustrates the chemical structures and the physical properties of some examples of compounds according to the invention. In the column "salt", "-" represents a compound in the form of the free base.

Cmpd. Structure Name Charact.

Salt

Number

(8R)-8-tert- MS (method D): m/z

Butyl- = 229.17 (ES+);

6,7,8,9- tetrahydro- ¹ H NMR(300 MHz,

5H- DMSO-de) δ 11.08 (s, pyrido[4,3- 1H), 8.63 (s, 1H), bjindole

8.06 (d, 1H), 7.20 (d,

9 1H), 2.82-2.73 (m,

\

H 3H), 2.25 (m, 1H),

2.07 (m, 1H), 1.45 (m, 2H), 0.98 (s, 9H). Absolute

configuration not confirmed.

(8S)-8-tert- MS (method D): m/z

Butyl- = 229.19 (ES+);

6,7,8,9- tetrahydro- ¹ H NMR(300 MHz,

5H- CDCI ₃ )512.10 (br, pyrido[4,3- 1H), 8.52 (s, 1H), bjindole

\ 7.91 (s, 1H), 7.50 (s,

H

10 1H), 2.99-2.68 (m,

3H), 2.38 (m, 1H), 2.12 (m, 1H), 1.48 (m, 2H), 0.98 (s, 9H). Absolute

configuration not confirmed.

Cmpd. Structure Name Charact.

Salt

Number

8-Phenyl- MS (method A): m/z 6,7,8,9- = 249.15 (ES+); RT = tetrahydro- 5H- 2.57 minutes.

pyrido[4,3-

20 b]indole -

H

8-(1 ,1 - MS (method A): m/z

Dimethyl- = 243.18 (ES+). propyl)-

6,7,8,9-

21 tetrahydro- -

5H- pyrido[4,3- b]indole

H

8-Pentyl- MS (method A): m/z 6,7,8,9- = 243.19 (ES+). tetrahydro- 5H- pyrido[4,3- bjindole

22 -

H

8-tert-Butyl- MS (method F): m/z

1 -methyl- = 243.15 (ES+); RT =

6,7,8,9-tetra

hydro-5H- 3.07 minutes.

23 pyrido[4,3- - b]indole

H Cmpd. Structure Name Charact.

Salt

Number

2,3,4,4a,5,6 MS (method A): m/z

,7,1 1 c- = 227.16 (ES+)

Octahydro-

1 H-ben

24 zo[e]pyrido[ -

4,3-b]indole

H

Biological Examples

The pharmacological properties of the compounds of the invention may be confirmed by a number of pharmacological assays. The exemplified assays which follow have been carried out with compounds according to the invention.

The Eg5 pharmacological assay is conducted using purified recombinant human kinesin motor domain protein (Cytoskeleton Inc. Catalog number EG01 -XL ). The ATPase activity of Eg5 in the presence of preformed tubulin is used as a measure of the function of Eg5. Briefly, Eg5 protein is added to the assay buffer (Cytoskeleton Inc. Catalog number BK060), which contains 15mM PIPES, pH 7.5, 5mM MgCI ₂ , preformed tubulin, 2-amino-6-mercapto-7-methylpurine ribonudeoside and purine nucleoside phosphorylase. The mixture was incubated for 20 minutes in the presence and absence (as control) of the test compound at 22°C following the addition of ATP to a final concentration of 0.62mM. The release of inorganic phosphate from ATP hydrolyzed by the ATPase activity of Eg5 is monitored by an absorbance shift (330 - 360 nm) when 2-amino-6-mercapto-7-methylpurine ribonudeoside is catalytically converted to 2-amino-6-mercapto-7-methyl purine in the presence of inorganic phosphate by purine nucleoside phosphorylase. The inhibitory effect of the test compound is measured by the decrease in rate of OD ₃₆ o shift relative to that in the absence of the compound.

The cell viability assay is conducted using various hematological malignant cell lines and the results are exemplified using a non-Hodgkin lymphoma cell line of WSU- DLCL2. Cells are suspended to a density of 0.1 million/ml in culture media of RPMI- 1640 supplemented with 25 mM HEPES, L-glutamine, 10% heat-inactivated fetal bovine serum. Triplicates of cell suspension were added to a 96-well plate and were incubated for 72 hours in the presence and absence of test compounds at different concentrations. At the end of incubation, cell viability is determined by MTS reagent (Cell Titer 96 Aqueous, catalog number G358B) supplied by Promega Corporation. The effect of the test compound on cell viability at each compound concentration is expressed as percent of cell viability measured in the absence of the compound. The IC50 of the test compound was calculated from the dose response curve of the test compound.

Generally the compounds of this invention exhibit potent Eg5 and viability inhibitory activities. Results are provided below in Table 1 .

- 55 -

>3 >10

H

>3 >10

H

The efficacy of the compounds of formula (I) of this invention in treating a variety of diseases as disclosed herein can be confirmed by any of the methods known to one skilled in the art. For instance, the efficacy in treating cancer can be confirmed by inhibiting and/or affecting the growth of tumor cells implanted in immuno-deficient mice (SCID) according to methods well known to those skilled in the art.