Background art.

Interest in FTase inhibitors as agents for cancer chemotherapy was initiated by the observation that the protein product of the ras oncogene required farnesylation for membrane binding and biological function. Indeed, many FTase inhibitors have shown growth inhibition on a variety of human tumor cell lines and, more recently, some of them have exhibited clinical efficacy for cancer patients.

Interestingly, the original hypothesis that FTase inhibitors would block Ras function has been proved invalid by recent studies. It is now generally acknowledged that the biological effects and clinical efficacy of FTase inhibitors must be due to the inhibition of the farnesylation of other proteins (Review: Karp J.

E. , et al, Current Opinion in Oncology, 2001,13, 470- 476).

About 20 small membrane-bound GTP binding proteins such as H-Ras, K-Ras, N-Ras, RhoB, RhoE, lamin A, lamin B, Rap 2, HDJ-2, CENP-E, CENP-F, PxF, cGMP phosphodiesterase, phosphorylase A, phosphorylase B, tyrosine phosphatase, transducin, rhodopsin kinase, possessing the farnesyl group have been identified and it has been reported that

many of them act as molecular switches to regulate cell cycle, growth, proliferation, differentiation, and/or apoptosis (Review: Tamanoi F. , et al, Journal of Cellular Biochemistry, 2001,64-70).

In tumor cells, the constitutive activation of some of those proteins contributes to their malignant growth properties. In normal cells, on the other hand, this switching mechanism is highly regulated and those proteins are found bound predominantly in their inactive GDP binding state.

FTase farnesylates CAAX-containing proteins (C, Cys ; A, an aliphatic amino acid; X, Ser, Met, Cys, Gln, or Ala). The present inventors have identified about 400 peptides that have a CAAX tetrapeptide sequence in their C-terminal in human genes. And now it is being investigated which peptide has a correlation between the biological effects and efficacy in animal models using DNA array and proteomics technologies.

Summary of the invention It is an object of the present invention to provide more potent compounds for the treatment of abnormal cell growth, including cancers, in a mammal by inhibiting farnesyl-protein transferase (FTase). In particular, the present invention relates to a novel benzofuran compounds of the formula [I],

wherein: R1 is-NO2i-CN ; or pyridine-3-yl; R2 is hydrogen; halogen; phenyl which may be optionally substituted with, each the same or different, 1 to 3 halogen, -CN, or straight or branched (C1-C4)-alkoxy ; pyridine-3-yl; or benzo [1, 3] dioxole-5-yl. and pharmaceutically acceptable salts and solvates thereof.

It is another object of the present invention to provide pharmaceutical compositions containing these compounds, and the use of those compounds in the medical therapy, especially in treatment of tumor.

A still further object of the present invention is to provide a process for the preparation of those compounds.

Unless otherwise indicated, the following definitions are set forth to illustrate and define the meaning and scope of the various terms used to describe the present invention herein.

The term"halogen"refers to fluoro, chloro, bromo, and iodo.

The term"alkoxy"refers to the group-O-R wherein R is alkyl, a straight chain or a branched chain of hydrocarbon group containing a maximum of 4 carbon atoms., e. g. , methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, iso-butoxy, sec-butoxy, and tert-butoxy.

In the present invention, the expression"substituted by"means that the substitution can occur at one or more positions and, unless otherwise indicated, that the

substituents are independently selected from the specified options.

"Pharmaceutically acceptable salt"refers to conventional acid-addition salts or base-addition salts that retain the biological effectiveness and properties of the compounds of formula (I) and are formed from suitable non-toxic organic or inorganic acids or organic or inorganic bases. Sample acid-addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, and those derived from organic acids such as p-toluenesulfonic acid, salicylic acid, methanesulfonic acid, oxalic acid, succinic acid, citric acid, malic acid, lactic acid, fumaric acid, and the like. Sample base-addition salts include those derived from potassium, sodium, ammonium, and quarternary ammonium hydroxide, such as for example tetramethylammonium hydroxide. The term"pharmaceutically acceptable salt"includes prodrugs of compounds of formula (I).

"Pharmaceutically acceptable"such as pharmaceutically acceptable carrier, excipient, prodrug, etc. , means pharmacologically acceptable and substantially non-toxic to the subject to which the particular compound is administered.

"Pharmaceutically active metabolite"means a metabolic product of a compound of formula (I) that is pharmaceutically acceptable and effective.

The term"prodrug"refers to a compound of the formula (I) that may be converted under physiological conditions or by solvolysis to any of the compounds of formula (I) or to a pharmaceutically acceptable salt of compounds of formula (I). A prodrug may be inactive when administered

to a subject but is converted in vivo to an active compound of formula (I).

The compounds of formula I can also be solvated, e. g. hydrated. The solvation can be effected in the course of the manufacturing process or can take place e. g. as a consequence of hygroscopic properties of an initially anhydrous compound of formula I (hydration). The term pharmaceutically acceptable salts also includes physiologically acceptable solvates.

The present invention relates to novel benzofuran compounds of formula (I), and pharmaceutically acceptable salts, prodrugs and solvates thereof.

Best mode for carrying out the invention The present invention concerns benzofuran compounds of the formula [I], wherein: R1 is-NO2 ; -CN; or pyridine-3-yl; R2 is hydrogen; halogen; phenyl which may be optionally substituted with, each the same or different, 1 to 3 halogen, -CN, or straight or branched (C1-C4)-alkoxy ; pyridine-3-yl; or benzo [1, 3] dioxole-5-yl; and pharmaceutically acceptable salts or solvates thereof.

In a preferred embodiment, the invention comprises benzofuran compounds of formula (I), wherein R2 is hydrogen, halogen, phenyl, phenyl substituted with, each the same or different, 1 to 3 halogen, -CN, methoxy or ethoxy groups, pyridine-3-yl, or benzo [1, 3] dioxole-5-yl.

Especially preferred are compounds of formula (I), wherein R2 is phenyl, phenyl substituted with, each the same or different, 1 to 3 fluoro, bromo, -CN, methoxy or ethoxy groups, pyridine-3-yl, or benzo [1, 3] dioxole-5-yl.

Even more preferred are compounds of formula (I), wherein R2 is 3-fluorophenyl, 4-fluorophenyl, 3,4- difluorophenyl, 3,5-difluorophenyl, 3-chlorophenyl, 3- cyanophenyl, 3-methoxyphenyl, or 3-ethoxyphenyl.

In a further preferred embodiment, the invention comprises compounds of formula (I), wherein R1 ;-CN ; or pyridine-3-yl; and R2 is halogen, phenyl, 3-chlorophenyl, 3-fluorophenyl, 3- methoxyphenyl, 3-ethoxyphenyl, 3-cyanophenyl, pyridine-3- yl, or benzo [1, 3] dioxole-5-yl.

In another preferred embodiment, the invention comprises compounds of the formula (I), wherein Rl is-NO2 ; and R2 is hydrogen ; halogen; phenyl which may be optionally substituted with, each the same or different, 1 to 3 halogen, -CN, or straight or branched (Cl-C4)-alkoxy ; pyridine-3-yl; or benzo [1, 3] dioxole-5-yl.

Preferred compounds in accordance with the present invention are as follows: a) 4- [Amino- (3-methyl-3H-imidazol-4-yl)- (5-nitro-7-

phenyl-benzofuran-2-yl)-methyl]-benzonitrile b) 4- [Amino- [7- (3-methoxy-phenyl)-5-nitro-benzofuran- 2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile ; c) 4- [Amino- (7-benzo [1, 3] dioxol-5-yl-5-nitro- benzofuran-2-yl)- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile ; d) 4- [Amino- [7- (3-fluoro-phenyl)-5-nitro-benzofuran-2- yl]- (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile ; e) 4- [Amino- [7- (3-cyano-phenyl)-5-nitro-benzofuran-2- yl]- (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile ; f) 4- [Amino- [7- (3-ethoxy-phenyl)-5-nitro-benzofuran-2- yl]- (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile ; g) 4- [Amino- [7- (3-chloro-phenyl)-5-nitro-benzofuran-2- yl]- (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile ; h) 4- [Amino- [7- (4-fluoro-phenyl)-5-nitro-benzofuran-2- yl]- (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile ; i) 4- [Amino- [7- (3, 5-difluoro-phenyl) -5-nitro- benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile; j) 4- [Amino- [7- (3, 4-difluoro-phenyl) -5-nitro- benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile; k) 4- [Amino- (3-methyl-3H-imidazol-4-yl)- (5-nitro-7- pyridin-3-yl-benzofuran-2-yl) -methyl] -benzonitrile ; and <BR> <BR> <BR> <BR> <BR> <BR> <BR> 1) 4- [Amino- (3-methyl-3H-imidazol-4-yl)- (5-nitro-<BR> <BR> <BR> <BR> <BR> benzofuran-2-yl)-methyl]-benzonitrile.

In another preferred embodiment, the present invention comprises compounds of formula (I) wherein

R'is-CN ; and R2 is hydrogen; halogen; phenyl which may be optionally substituted with, each the same or different, 1 to 3 halogen,-CN, or straight or branched (Cl-C4)-alkoxy ; pyridine-3-yl; or benzo [1, 3] dioxole-5-yl.

Preferred benzofuran compounds in accordance with the present invention are as follows: a) 2- [Amino- (4-cyano-phenyl)- (3-methyl-3H-imidazol-4- yl)-methyl]-7-phenyl-benzof. uran-5-carbonitrile ; b) 2- [Amino- (4-cyano-phenyl)- (3-methyl-3H-imidazol-4- yl)-methyl]-7- (3-methoxy-phenyl)-benzofuran-5- carbonitrile; c) 2- [Amino- (4-cyano-phenyl)- (3-methyl-3H-imidazol-4- yl)-methyl]-7-benzo [1,3] dioxol-5-yl-benzofuran-5- carbonitrile; d) 2- [Amino- (4-cyano-phenyl)- (3-methyl-3H-imidazol-4- yl)-methyl]-7- (3-fluoro-phenyl)-benzofuran-5-carbonitrile ; e) 2- [Amino- (4-cyano-phenyl)- (3-methyl-3H-imidazol-4- yl)-methyl]-7- (3-cyano-phenyl)-benzofuran-5-carbonitrile ; f) 2- [Amino- (4-cyano-phenyl)- (3-methyl-3H-imidazol-4- yl)-methyl]-7- (3-chloro-phenyl)-benzofuran-5-carbonitrile ; and g) 2- [Amino- (4-cyano-phenyl)- (3-methyl-3H-imidazol-4- yl)-methyl]-7-pyridin-3-yl-benzofuran-5-carbonitrile.

In a further preferred embodiment, the present invention comprises compounds of formula (I) wherein R1 is pyridine-3-yl; and R2 is hydrogen; halogen; phenyl which may be optionally

substituted with, each the same or different, 1 to 3 halogen, -CN, or straight or branched (C1-C4)-alkoxy ; pyridine-3-yl; or benzo [1, 3] dioxole-5-yl.

Preferred benzofuran compounds in accordance with the present invention are as follows: a) 4- [Amino- [7- (3-fluoro-phenyl)-5-pyridin-3-yl- benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile ; b) 4- [Amino- (3-methyl-3H-imidazol-4-yl)- (7-phenyl-5- pyridin-3-yl-benzofuran-2-yl) -methyl] -benzonitrile ; c) 4- [Amino- [7- (3-methoxy-phenyl)-5-pyridin-3-yl- benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile ; d) 4- [Amino- (7-benzo [1, 3] dioxol-5-yl-5-pyridin-3-yl- benzofuran-2-yl)- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile; e) 4- [Amino- [7- (3-cyano-phenyl)-5-pyridin-3-yl- benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile ; f) 4- [Amino- [7- (3-chloro-phenyl)-5-pyridin-3-yl- benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile ; and g) 4- [Amino- (5, 7-di-pyridin-3-yl-benzofuran-2-yl)- (3- methyl-3H-imidazol-4-yl)-methyl]-benzonitrile.

The benzofuran compounds of the formula (I) show strong anti-tumor activity against various tumor cell lines. This anti-tumor activity indicates that the compounds of the formula (I) and pharmaceutically acceptable salts thereof can be used as anti-tumor agents.

In another embodiment, the present invention concerns pharmaceutical compositions comprising a benzofuran compound of formula (I) and a pharmaceutically acceptable carrier.

In a preferred embodiment, the present invention concerns a pharmaceutical composition comprising benzofuran compound of formula (I) and chemotherapeutic.

Preferably, the pharmaceutical compositions according to the present invention are useful for the treatment of cell proliferative disorders, preferably for cancer, and more preferably for colorectal cancer, lung cancer, breast cancer, prostate cancer and leukemia.

The benzofuran compounds of the formula (I) are active against a variety of cell lines including colorectal cancer, lung cancer, breast cancer, prostate cancer, and leukemia.

In another embodiment, the present invention comprises the use of the benzofuran compounds of formula (I) for the preparation of medicaments, especially for the preparation of medicaments for the treatment of cell proliferative disorders.

More preferably the use according to the present invention is for the preparation of medicaments for the treatment of cancer and most preferably colorectal cancer, lung cancer, breast cancer, prostate cancer, and leukemia.

For clinical use, the benzofuran compounds of the formula (I), their salt forms thereof and the like can be administered alone, but will generally be administered in pharmaceutical admixture formulated as appropriate to the particular use and purpose desired, by mixing excipient, binding agent, lubricant, disintegrating agent, coating material, emulsifier, suspending agent, solvent,

stabilizer, absorption enhancer and/or ointment base. The admixture can be used for oral, injectable, rectal or topical administration.

In more detail, as mentioned earlier, medicaments containing a compound of formula (I) or its prodrug are also an object of the present invention, as is a process for the manufacture of such medicaments, which process comprises bringing one or more compounds of formula (I) and, if desired, one or more other therapeutically valuable substances into a galenical administration form.

This invention also relates to a pharmaceutical composition for inhibiting abnormal cell growth in a mammal, including a human, comprising an amount of a benzofuran compound of the formula (I) as defined above, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, that is effective in inhibiting Farnesyl-protein Transferase, and a pharmaceutically acceptable carrier.

The pharmaceutical composition according to the present invention comprises a therapeutically effective amount of a benzofuran compound of the formula (I) as defined above, or a pharmaceutically acceptable salt, prodrug, or solvate thereof and, and a pharmaceutically acceptable carrier, and, if desired, a chemotherapeutic.

In one embodiment, the chemotherapeutic is selected from the group consisting of mitotic inhibitors, alkylating agents, anti-metabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormones, and anti-androgens.

The pharmaceutical composition according to the present invention comprising an amount of a benzofuran derivative of the formula (I), a. pharmaceutically acceptable salt, prodrug, or solvate of such compound,

that is effective in inhibiting Farnesyl-protein Transferase, is useful for the treatment of mammal disorders, as for example lung cancer, NSCLC (non small cell lung cancer), bone cancer, pancreatic cancer, skin cancer, cancer of the head and neck, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, gynaecologic tumors (e. g., uterine sarcoma, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina or carcinoma of the vulva), Hodgkin's Disease, cancer of the oesophagus, cancer of the small intestine, cancer of the endocrine system (e. g., cancer of the thyroid, parathyroid or adrenal glands), sarcomas of soft tissues, cancer of the urethra, cancer of the penis, prostate cancer, chronic or acute leukemia, solid tumors of childhood, lymphocytic lymphomas, cancer of the bladder, cancer of the kidney or ureter (e. g., renal cell carcinoma, carcinoma of the renal pelvis, paediatric, malignancy, neoplasms of the central nervous system, (e. g. , primary CNS lymphoma, spinal axis tumor, brain stem gliomas or pituitary adenomas), Barrett's esophagus (pre-malignant syndrome), neoplastic cutaneous disease, psoriasis, mycoses fungoides, benign prostatic hypertrophy, human papilloma virus (HPV), and restinosis.

More specifically, the pharmaceutical composition according to the present invention is useful for the treatment of colorectal cancer, lung cancer, breast cancer, prostate cancer, and leukemia.

The pharmaceutical compositions may be administered orally, for example in the form of tablets, coated tablets, dragees, hard or soft gelatine capsules, solutions, emulsions or suspensions. Administration can also be carried out rectally, for example using

suppositories; locally or percutaneously, for example using ointments, creams, gels or solutions; or parenterally, for example using injectable solutions.

For the preparation of tablets, coated tablets, dragees or hard gelatine capsules the compounds of the present invention may be admixed with pharmaceutically inert, inorganic or organic excipients. Examples of suitable excipients for tablets, dragees or hard gelatine capsules include lactose, maize starch or derivatives thereof, talc or stearic acid or salts thereof.

Suitable excipients for use with soft gelatine capsules include for example vegetable oils, waxes, fats, semi-solid or liquid polyols etc.; according to the nature of the active ingredients it may however be the case that no excipient is needed at all for soft gelatine capsules.

For the preparation of solutions and syrups, excipients which may be used include for example water, polyols, saccharose, invert sugar and glucose.

For injectable solutions, excipients which may be used include for example water, alcohols, polyols, glycerine, and vegetable oils.

For suppositories, and local or percutaneous application, excipients which may be used include for example natural or hardened oils, waxes, fats and semi- solid or liquid polyols.

The pharmaceutical compositions may also contain preserving agents, solubilizing agents, stabilizing agents, wetting agents, emulsifiers, sweeteners, colorants, odorants, salts for the variation of osmotic pressure, buffers, coating agents or antioxidants. They may also contain other therapeutically valuable agents.

In summary, a pharmaceutical formulation for oral administration may be granule, tablet, sugar coated tablet, capsule, pill, suspension or emulsion. A pharmaceutical formulation for parenteral injection, for example, intravenously, intramuscularly or subcutaneously, may be used in the form of a sterile aqueous solution which may contain other substances, for example, salts or glucose to make the solution isotonic. The anti-tumor agent can also be administered in the form of a suppository or pessary, or they may be applied topically in the form of a lotion, solution, cream, ointment or dusting powder.

The daily dosage level of the benzofuran compounds of the formula (I) is from 5 to 2,000 mg/m2 when administered by either the oral or parenteral route. Thus tablets or capsules can contain from 5 mg to 1,000 mg of active compound for administration singly or two or more at a time as appropriate. In any event the actual dosage can be dependent on the weight and response of the particular patient.

The present invention relates also to a method for treating cell proliferative disorders comprising administering to a patient in need thereof a therapeutically effective amount of benzofuran compound of the formula (I). The present invention concerns specifically a method for treating cancer, and, more specifically, colorectal cancer, lung cancer, breast cancer, prostate cancer, and leukemia.

The present invention also relates to methods of inhibiting abnormal cell growth, including cancer, in a mammal, including human, comprising administering to said mammal an amount of a benzofuran compound of the formula (I) as defined above, or a pharmaceutically acceptable salt, prodrug, solvate thereof, that is effective in

inhibiting Farnesyl-protein Transferase.

The present invention also relates to methods of inhibiting abnormal cell growth, including cancer, in a mammal, including human, comprising administering to said mammal an amount of a benzofuran compound of the formula (I) as defined above, or a pharmaceutically acceptable salt, prodrug, solvate thereof, that is effective in inhibiting abnormal cell growth.

The present invention also relates to methods of inhibiting abnormal cell growth, including cancer, in a mammal, including human, comprising administering to said mammal an amount of a benzofuran compound of the formula (I) as defined above, or a pharmaceutically acceptable salt, prodrug, solvate thereof, that is effective in combination with a chemotherapeutic. In one embodiment, the chemotherapeutic is selected from the group consisting of mitotic inhibitors, alkylating agents, anti- metabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormones, and anti-androgens.

The present invention further relates to methods of inhibiting abnormal cell growth, including cancer, in a mammal, including human, comprising administering to said mammal an amount of a benzofuran compound of the formula (I) as defined above, or a pharmaceutically acceptable salt, prodrug, solvate thereof, that is effective in combination with radiation therapy, wherein the amount of the compound, salt, prodrug, or solvate of a benzofuran compound the formula (I) is in combination with the radiation therapy effective in inhibiting abnormal cell growth in the mammal. Techniques for administering radiation therapy are known in the art, and these techniques can be used in the combination therapy

described herein. The administration of the compound of the invention in this combination therapy can be determined as described herein.

The benzofuran compound of the formula (I) can render abnormal cells more sensitive to treatment with radiation for purposes of killing and/or inhibiting the growth of such cells. Accordingly, this invention further relates to a method for sensitizing abnormal cells in a mammal to treatment with radiation which comprises administering to the mammal an amount of a benzofuran compound of the formula (I) or a pharmaceutically acceptable salt, prodrug, or solvate thereof, which amount is effective in sensitizing abnormal cells to treatment with radiation.

The amount of the compound, salt, prodrug, or solvate in this method can be determined according to the means for ascertaining effective amounts of such compounds described herein.

Benzofuran compounds of the formula (I) of the present invention can be produced via the following process: A process for producing benzofuran compounds of the formula [I],

wherein Rl and R2 are the same as above defined, which comprises cleaving a compound of formula [VII], 4- <BR> <BR> <BR> [2-methyl-2-propanesulfinamide-benzofuran-2-yl- (3-methyl- 3H-imidazol-4-yl)-methyl]-benzonitrile,

wherein R1 and R2 are the same as above defined, on the 2-methyl-2-propanesulfine group by the treatment with an acidic compound.

Further, the present invention includes a process for producing the 4- [2-methyl-2-propanesulfinamide-benzofuran- 2-yl- (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile compound of the formula [VII],

wherein R1 and R2 are the same as above defined, which comprises alkylating a 2-methyl-propane-2-sulfinic acid benzofuran-2-yl- (4-cyano-phenyl)-methyleneamide compound of the formula [V], wherein R1 and R2 are the same as above defined, by 1-methyl-2-triethylsilanyl-lH-imidazole-5-lithium of the formula [VIII].

Another object of the present invention is a process for producing the 2-methyl-propane-2-sulfinic acid benzofuran-2-yl- (4-cyano-phenyl)-methyleneamide compound of the formula [V],

wherein R1 and R2 are the same as above defined, which comprises condensing a 4- (benzofuran-2-carbonyl)- benzonitrile compound of the formula [II], wherein R1 and R2 are the same as above defined, with 2-methyl-2-propanesulfinamide [VI],

in the presence of a Lewis acidic dehydrating agents.

Further the present invention concerns a process for producing the 4- (benzofuran-2-carbonyl)-benzonitrile compound of the formula (II),

wherein R1 and R2 are the same as above defined, which comprises condensing a 2-hydroxybenzaldehyde compound of the formula [III], wherein R1 and R2 are the same as above defined, with a 4-cyanophenacyl halide of the formula [IV],

wherein X is halide, in the presence of a basic compound.

The following examples illustrate the preferred methods for the preparation of the compounds of the present invention, which are not intended to limit the scope of the invention thereto.

The physical data of the compounds were recorded as a mixture of diastereomers regarding the chiral center of the carbon atom unless otherwise noted.

The retention time of each compound in LCMS was recorded using the following method unless otherwise noted.

Condition A: column: Inertsil ODS-3/4. 0x33 mm (GL Science Inc.) mobile phase: 0. 05% TFA-water : 0. 05% TFA- acetonitrile, flow rate: 1.0 ml/min gradient: 10% MeCN at 0 min-+ 95% MeCN at 4 min 95% MeCN at 5.5 min-"10%. MeCN at 6.0 min Condition B: column: WakopakX Combi ODS/4. 0x50 mm (Wako Inc.) mobile phase: 0. 05% TFA-water : 0. 05% TFA-

acetonitrile, flow rate: 4.0 ml/min gradient: 10% MeCN at 0 min-+ 95% MeCN at 3.5 min 10% MeCN at 4.5 min--10% MeCN at 5.0 min The requisite starting materials and some intermediates are in some cases commercially available, or can be prepared according to literature procedures.

In the following schemes and examples,"Me"represents a methyl moiety and"Et"represents an ethyl moiety.

Hence, for example,"HOEt"means ethanol. Also"THF"means <BR> <BR> <BR> tetrahydrofuran,"DCM"means dichloromethane,"DMSO"means dimethylsuloxide,"DMF"means N, N-dimethylformamide, EtOAc means ethyl acetate, MeCN means acetonotrile, LDA means lithium diisopropylamide, WSCI means 1- (3- dimethylaminopropyl) -3-ethyl-carbodiimide hydrochloride, "DIEA"means N-ethyldiisopropylamine, HOBt means N- hydroxybenzotriazole monohydrate, TFA means trifluoroacetic acid, DMAP means 4-dimethylaminopyridine, "HMT"means hexamethylenetetramine, and Pd (PPh3) 4 means tetrakis (triphenylphosphine) -palladium (0).

Examples Process 1 Preparation of 4- [amino- (3-methyl-3H-imidazol-4-yl)- (5- nitro-benzofuran-2-yl)-methyl]-benzonitrile derivatives A series of 4- [amino- (3-methyl-3H-imidazol-4-yl)- (5- nitro-benzofuran-2-yl) -methyl] -benzonitrile derivatives 9 were prepared as shown in Scheme 1.

Coupling of commercially available p-cyanophenacyl

bromide 1 and commercially available 3-bromo-5-nitro- salicylaldehyde 2, in the presence of potassium carbonate in acetonitrile under reflux, leaded to keto-benzofuran 3 in high yield according to literature procedure (Stille, J. R. , Ward, J. A. , Leffelman, C. , and Sullivan, K. A., Tetrahedron Letters, 1996,37 (52), 9353-9356).

The sulfine imine 5 was prepared by tetraethyl orthotitanate mediated condensation of keto-benzofuran 3 with commercially available (R) -or (S) -sulfineamide 4 following the literature procedure (Liu, G. , Cogan, D. A., Owens, T. D. , Tang, T. P. , and Ellman, J. A. , J. Org.

Chem. 1999,64, 1278-1284).

Alkylation of sulfine imine 5 was achieved with 2- trimethylsilyl-5-lithium-N-methylimidazole 6 which was prepared by according to Carpenter's procedure (Carpenter, A. J. , and Chadwick, D. J., Ttetrahedron, 1986,42, 2358) at tetrahydrofurane at-78 degree for 10 minutes to give a key intermediate sulfineamide 7.

A variety of aryl substituents was introduced into the key intermediate sulfineamide 7 by the cross-coupling reaction with arylboronic acid or arylboronate in the presence of catalytic amount of palladium complex under Suzuki reaction conditions to give biaryl 8 followed by deprotection of the tert-butylsulfine group under acidic conditions yielded desired 4- [amino- (3-methyl-3H-imidazol- 4-yl)- (5-nitro-benzofuran-2-yl)-methyl]-benzonitrile derivatives 9.

Scheme 1-Preparation of 5-nitro benzofuran KZC03 H N02 CH3CN H NOz CH3CN i Br + HO I I O/ NOz N Br Br 2 3 son 0 *Sl Et S' Ti (OEt) 4 1 Et3S + NH2 THF-N reflux 0 N02 + Br 4 5 6 1--N/-- r 9 (OH) 2 , N THF N-S,-o Pd (ph,) 4 XN-S>o -780C THF I 0-N02 reflux 0 N02 i N v -r z Br R2 7 8 zon N/ HCI gNH2 THE rt 0 N02 R2 9 9

Process 2 Preparation of 4- [amino- (3-methyl-3H-imidazol-4-yl)- (5- cyano-benzofuran-2-yl)-methyl]-benzonitrile derivatives A series of 4- [amino- (3-methyl-3H-imidazol-4-yl)- (5- cyano-benzofuran-2-yl) -methyl] -benzonitrile derivatives 16 were prepared as shown in Scheme 2.3-Formyl-4-hydroxy- benzonitrile 10 was prepared according to literature procedure (Suzuki, Y. et al. Chem. Pharm. Bull. 1983, 31, 1751-1753). Bromination of 3-formyl-4-hydroxy-benzonitrile 10 was achieved by the treatment with bromine in ethanol at room temperature to give 3-bromo-5-formyl-4-hydroxy- benzonitrile 11.

Coupling of commercially available p-cyanophenacyl bromide 1 and 3-bromo-5-formyl-4-hydroxy-benzonitrile 11 in the presence of potassium carbonate in acetonitrile under reflux, leaded to keto-benzofuran 12 in high yield.

The sulfine imine 13 was prepared by tetraethyl orthotitanate mediated condensation of keto-benzofuran 12 with sulfineamide 4.

Alkylation of sulfineimine 13 was achieved with 2- trimethylsilyl-5-lithium-N-methylimidazole 6 at tetrahydrofurane at-78 degree for 10 minutes to give a key intermediate'sulfineamide 14.

A variety of aryl substituents was introduced into the key intermediate sulfineamide 14 by the cross-coupling reaction with arylboronic acid or boronate in the presence of catalytic amount of palladium complex under Suzuki reaction conditions to give biaryl 15 followed by deprotection of the tert-butylsulfine group under acidic conditions yielded desired 4- [amino- (3-methyl-3H-imidazol- 4-yl)- (5-cyano-benzofuran-2-yl)-methyl]-benzonitrile derivatives 16.

Scheme 2-Preparation of 5-cyano benzofuran derivatives Brz KZC03 H H CN EtOH H » CN 4 CH3CN HO J 1 reffux HO) : ; : r Br 10 11 Q\ w Ti (OEt) a I /O/\ CN, SNI-ia THF ', I Br N-) ==/ Br Br 12 4 13 /= \/ , C-. Et3si THF N-S,, o R2B (OR) 2 >-78oC Pd (PPh3) 4 Ii N P P Br CN THF ii Br reflux bu 6 14 /=N \//=N Nr W Nr- IN-Sx HCI XNH2 Tuf O/\ CN rt /O/\ CN N-- / R2 R2 15 16

Process 3 Preparation of 4- [amino- (3-methyl-3H-imidazol-4-yl)- (5- pyridin-3-yl-benzofuran-2-yl)-methyl]-benzonitrile derivatives A series of 4- [amino- (3-methyl-3H-imidazol-4-yl)- (5- pyridin-3-yl-benzofuran-2-yl)-methyl]-benzonitrile derivatives 23 were prepared as shown in Scheme 3.2- Hydroxy-5-pyridin-3-yl-benzaldehyde was prepared according to literature procedure (Johnson, Roy A. , Nidy, Eldon G., Aiken, James W. , Crittenden, Norman J., Gorman, Robert R., J. Med. Chem., (1986), 29, 1461-1468). Bromination of 2- hydroxy-5-pyridin-3-yl-benzaldehyde was preformed by the treatment with benzyltrimethylammonium tribromide in a mixed solution of DCM and methanol at room temperature to give 3-bromo-2-hydroxy-5-pyridin-3-yl-benaldehyde 18.

Coupling of p-cyanophenacyl bromide 1 and 3-bromo-2- hydroxy-5-pyridin-3-yl-benzaldehyde 18 in the presence of potassium carbonate in acetonitrile under reflux, leaded to keto-benzofuran 19 in high yield. The sulfine imine 20 was prepared by tetraethyl orthotitanate mediated condensation of keto-benzofuran 19 with sulfineamide.

Alkylation of sulfineimine 20 was achieved with 2- trimethylsilyl-5-lithium-N-methylimidazole 6 to give a key intermediate sulfineamide 21.

A variety of aryl substituents was introduced into the key intermediate sulfineamide 21 by the cross-coupling reaction with arylboronic acid or boronate in the presence of catalytic amount of palladium complex under Suzuki reaction conditions to give biaryl 22 followed by deprotection of the tert-butylsulfine group under acidic conditions yielded desired 4- [amino- (3-methyl-3H-imidazol- 4-yl)- (5-pyridin-3-yl-benzofuran-2-yl)-methyl]- benzonitrile derivatives 23.

Scheme 3 Preparation of 5-(pyridin-3-yl) benzofuran derivatives N I BTMABr3 N I KZC03 H DCM/MeOH H CH3CN I rt + Br 600C HO HO nU ** HO T Br 17 lg 1 O Ti (OEt) 4 -N THF , O/\ reffux - Ni \/+ NHz N 0/\ Br Br 19 4 20 N raz (OR) 2 * /Pd<'PPh _-N-Pd (PPh3) 4 N _78C \ THF -78oc \/-' -N Br 6 21 N o N Nw N4S LN-S HC1 TNH, tuf rut 0 N \/ R2 R2 22 23

The examples below provided are intended to assist in a further understanding of the invention. Particular materials employed species and conditions are intended to be further illustrative of the invention and not limitative of the reasonable scope thereof.

Of some benzofuran compound of the formula (I) the absolute stereochemical configuration was not experimentally determined. In those cases the stereochemically isomeric form which was first isolated is designed as"A"and the second as"B", without further reference to the actual stereochemical configuration.

Table 1: Example Ri R2 1-NO2-phenyl 2-NO2-3-methoxyphenyl 3-N02-benzo [1, 3] - dioxole-5-yl 4-N02-3-fluorophenyl 5-NO2-3-cyanophenyl 6-N02-3-ethoxyphenyl Example R1 R2 7-NO2-3-chlorophenyl 8-NO2-4-fluorophenyl 9 -NO2 -3, 5- difluorophenyl 10 -NO2 -3, 4- difluorophenyl 11-N02-pyridine-3-yl 12-N02-hydrogen 13-CN-phenyl 14-CN-3-methoxyphenyl 15-CN-benzo [1, 3] - dioxole-5-yl 16-CN-3-fluorophenyl 17-CN-3-cyanophenyl 18-CN-3-chlorophenyl 19-CN-pyridine-3-yl 20-pyridine-3-yl-3-fluorophenyl 21-pyridine-3-yl-phenyl 22-pyridine-3-yl-3-methoxyphenyl 23-pyridine-3-yl-benzo [1, 3]- dioxole-5-yl 24-pyridine-3-yl-3-cyanophenyl Example 25-pyridine-3-yl-3-chlorophenyl 26-pyridine-3-yl-pyridine-3-yl

Preparation of 4- [amino- (3-methyl-3H-imidazol-4-yl)- (5- nitro-benzofuran-2yl)-methyl]-benzonitrile compounds Example 1 Preparation of 4- [Amino- (3-methyl-3H-imidazol-4-yl)- (5- nitro-7-phenyl-benzofuran-2-yl)-methyl]-benzonitrile 1-a) Preparation of 4- (7-bromo-5-nitro-benzofuran-2- carbonyl) -benzonitrile, 3 To a solution of 3-bromo-2-hydroxy-5-nitro-benzaldehyde (15.0 g, 61.0 mmol) in CH3CN (300 ml) were added 4-cyano- phenacyl bromide (14.3 g, 64.0 mmol) and K2CO3 (10. 1 g, 73.2 mmol). The turbid mixture was heated to reflux for 4 hours. The mixture was cool to room temperature and filtered to collect a precipitate, and then the precipitate was washed with H20 and MeOH. The residue was dried in vacuo, and then 12.5 g of 4- (7-bromo-5-nitro-benzofuran-2- carbonyl) -benzonitrile was given as a light yellow solid.

LCMS (condition B), m/z [M+H] +, retention time minutes ; 1H- NMR (CDC13) d 8.68 (1H, d, J= 2. 0Hz), 8.64 (1H, d, J= 2. 0Hz), 8.28 (2H, dd, J=2. 0,6. 6 Hz), 7.91 (2H, dd, J=2.0, 6.6 Hz), 7. 86 (1H, s).

1-b) Preparation of 2-methyl-propane-2-sulfinic acid (7- bromo-5-nitro-benzofuran-2-yl)- (4-cyano-phenyl)- methyleneamide, 5 To a turbid solution of 4- (7-bromo-5-nitro-benzofuran- 2-carbonyl) -benzonitrile (11.0 g, 23.2 mmol) in THF (200

ml) were added (R) -or (S) -2-methyl-2-propane sulfinamide (3.09 g, 25.5 mmol) and tetraethyl orthotitanate (10.7 g, 46.4 mmol). After heating to reflux for 10 h, the mixture was cool to room temperature and poured into saturated NaCl (100 ml). The resulting mixture was filtered on celite pad and washed with AcOEt. The organic layer was separated, washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography over silica gel (eluent; CH2C12- > 10% MeOH in CH2Cl2) yielding 7.70 g of of 2-methyl- propane-2-sulfinic acid (7-bromo-5-nitro-benzofuran-2-yl)- (4-cyano-phenyl) -methyleneamide as an orange solid.

LCMS (condition B), m/z 474 [M+H] +, 476 [M+H] +, retention time 3.98 minutes; 1H-NMR (CDC13) d 8.52-8. 55 (2H, m), 7.78-7. 82 (3H, m), 7.68 (2H, m), 1.40 (9H, s).

1-c) Preparation of 2-methyl-propane-2-sulfinic acid [ (7- <BR> <BR> <BR> bromo-5-nitro-benzofuran-2-yl)- (4-cyano-phenyl)- (3-methyl- 3H-imidazol-4-yl) -methyl] -amide, 7 n-Butyllithium in hexane (1.58 M, 10.1 ml, 16.0 mmol) was added to a solution of N-methylimidazole (1.28 ml, 16.0 mmol) in THF (50 ml) at-78 °C. After stirring for 30 minutes, triethylsilyl chloride (2.69 ml, 16.00 mmol) was added to the reaction mixture. The mixture was allowed to warm to room temperature then cooled to-78 °C. n-Butyl- lithium in hexane (1.58 M, 10.1 ml, 16.0 mmol) was added again. The reaction mixture was allowed to warm to-20 °C and then cooled to-78 °C. The prepared mixture was added to a solution of 2-methyl-propane-2-sulfinic acid (7-bromo- 5-nitro-benzofuran-2-yl)- (4-cyano-phenyl)-methyleneamide (5.0 g, 10.5 mmol) in THF at-78 °C. After stirring for 10 minutes, saturated NHzCl was added. The reaction mixture was extracted with AcOEt. Combined organic layers were dried over Na2SO4 and evaporated in vacuo. The residue was purified by silica gel column chromatography over silica

gel (eluent; AcOEt-> 10% MeOH in AcOEt) yielding 3.64 g of 2-methyl-propane-2-sulfinic acid [ (7-bromo-5-nitro- <BR> <BR> <BR> <BR> benzofuran-2-yl)- (4-cyano-phenyl)- (3-methyl-3H-imidazol-4- yl) -methyl] -amide as an orange solid.

LCMS (condition B), m/z 556 [M+H] +, 558 [M+H] +, retention time 2.82 minutes ; 1H-NMR (CDC13) d 8.44 (1H, d, J=2.2 Hz), 8.39 (1H, d, J=2.2 Hz), 7.75 (2H, dd, J=2.0, 6.4 Hz), 7.64 (2H, dd, J=2.0, 6.4 Hz), 7.54 (1H, s), 7.28 (1H, s), 7.27 (1H, s), 6.95 (1H, s), 4.70 (1H, s), 3.15 (3H, s), 1.36 (9H, s).

1-d) Preparation of 2-methyl-propane-2-sulfinic acid [ (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)- (5-nitro-7- phenyl-benzofuran-2-yl) -methyl] -amide, 8 A mixture 2-methyl-propane-2-sulfinic acid [ (7-bromo-5- nitro-benzofuran-2-yl)- (4-cyano-phenyl)- (3-methyl-3H- imidazol-4-yl) -methyl] -amide (1.95 g, 3.50 mmol), 2-phenyl- [1, 3,2] dioxaborinane (1.70 g, 10.5 mmol), tetrakis (triphenylphosphine)-palladium (0) (1.21 g, 1.05 mmol) and tripotassium phosphate (2.23 g, 10.5 mmo) in THF (50 ml) was heated to reflux for 4 hours. The reaction mixture was diluted with AcOEt and then palladium was filtered off. The filtrate was washed with brine, dried over Na2SO4 and evaporated in vacuo. The residue was purified by column chromatography over silica gel (eluent; AcOEt-> 5 % MeOH in AcOEt-> 10 % MeOH in AcOEt), yielding 1.38 g of 2-methyl-propane-2-sulfinic acid [ (4-cyano- phenyl)- (3-methyl-3H-imidazol-4-yl)- (5-nitro-7-phenyl- benzofuran-2-yl) -methyl] -amide as a light yellow amorphous powder.

LCMS (condition B), m/z 554 [M+H] +, retention time 3.17 minutesi lH-NMR (CDCl3) d 8.46 (1H, d, J=2.3 Hz), 8.37 (1H, d, J=2.3 Hz), 7.72 (2H, dd, J=2.0, 6.6 Hz), 7.62-7. 66 (2H, m), 7.59 (2H, dd, J=2.0, 8.8 Hz), 7.53 (1H, s), 7.43-7. 51

(3H, m), 7.29 (1H, s), 6.90 (1H, s), 4.64 (1H, s), 3.14 (3H, s), 1.31 (9H, s).

1-e) Preparation of 4- [amino- (3-methyl-3H-imidazol-4-yl)- (5-nitro-7-phenyl-benzofuran-2-yl)-methyl]-benzonitrile To a solution of 2-methyl-propane-2-sulfinic acid [ (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)- (5-nitro-7- phenyl-benzofuran-2-yl) -methyl] -amide (50 mg, 0.09 mmol) in THF-methanol (3 ml, lml) at 0 °C was added 4M HC1 in AcOEt solution (1 ml). The reaction mixture was stirred at room temperature for 1 hour. The reaction was quenched with 1M NaOH aq (5 ml), and the resulting mixture was extracted with AcOEt. The organic layer was separated, washed with brine, dried over Na2SO4, filtered and concentrated in vacuo.

The residue was purified by column chromatography over silica gel (eluent; AcOEt-> 5 % MeOH in AcOEt-> 10 % MeOH in AcOEt), yielding 40 mg of 4- [amino- (3-methyl-3H- <BR> <BR> <BR> imidazol-4-yl)- (5-nitro-7-phenyl-benzofuran-2-yl)-methyl]- benzonitrile as a ligth yellow solid.

LCMS (condition B), m/z 451 [M+H] +, retention time 3.02 minutes 1H-NMR (CD30D) d 8.50 (1H, d, J=1.9 Hz), 8.36 (1H, d, J=1.9 Hz), 7.76-7. 80 (4H, m), 7.63-7. 65 (3H, m), 7.41- 7.51 (1H, m), 6.88 (1H, s), 6.57 (1H, s), 3.41 (3H, s) Example 2 <BR> <BR> <BR> <BR> <BR> <BR> 4- [Amino- [7- (3-methoxy-phenyl)-5-nitro-benzofuran-2-yl]- (3-<BR> <BR> <BR> <BR> <BR> methyl-3H-imidazol-4-yl)-methyl]-benzonitrile In a similar manner to that described in Example 1-d) and 1-e), the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-nitro-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3-methoxy-phenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in AcOEt) to give the titled compound as a colorless powder.

LCMS (condition B), m/z 480 [M+H] +, retention time 3.11 minutes ; 1H-NMR (CD30D) d 8.50 (1H, d, J=2.4 Hz), 8.35 (1H, d, J=2.4 Hz), 7.79 (2H, d, J=8.8 Hz), 7.63-7. 65 (4H, m), 7.40 (1H, dd, J=7.6, 7.6 Hz), 7.31-7. 33 (2H, m), 6.98-7. 01 (1H, m), 6.91 (1H, s), 6.61 (1H, s), 3. 78 (3H, s), 3. 43 (3H, s) Example 3 4- [Amino- (7-benzo [1, 3] dioxol-5-yl-5-nitro-benzofuran-2-yl)- (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile In a similar manner to that described in Example 1-d) and 1-e), the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-nitro-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 5- [1, 3,2] dioxaborinan-2-yl-benzo [1, 3] dioxole, followed by treatment with hydrogen chloride (4M solution in AcOEt) to give the titled compound as a colorless powder.

LCMS (condition B), m/z 494 [M+H] +, retention time 2.99 minutes ; lH-NMR (CDC13) d 8.35 (1H, d, J=2.3 Hz), 8.31 (1H, d, J=2.3 Hz), 7.75 (2H, d, J=8.6 Hz), 7.59 (2H, d, J=8.6 Hz), 7.78 (1H, s), 7.24 (1H, dd, J=2.0, 8.3 Hz), 7.22 (1H, d, J=2.0), 6.93 (1H, d, J=8.3 Hz), 6.65 (1H, s), 6.57 (1H, s), 6.05 (2H, s), 3.41 (3H, s) Example 4 <BR> <BR> <BR> <BR> <BR> <BR> <BR> 4- [Amino- [7- (3-fluoro-phenyl)-5-nitro-benzofuran-2-yl]- (3-<BR> <BR> <BR> <BR> <BR> <BR> methyl-3H-imidazol-4-yl)-methyl]-benzonitrile In a similar manner to that described in Example 1-d) and 1-e), the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-nitro-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3-fluoro-phenyl)- [1, 3,2] dioxaborinane, followed by

treatment with hydrogen chloride (4M solution in AcOEt) to give the titled compound as a colorless powder.

LCMS (condition B), m/z 468 [M+H] +, retention time 3.03 minutes ; 1H-NMR (CD30D) d 8.56 (1H, d, J=2.0 Hz), 8.42 (1H, d, J=2.0 Hz), 7.80 (2H, d, 8.8 Hz), 7.62-7. 65 (4H, m), 7.50-7. 56 (2H, m), 7.20 (1H, ddd, J=2.0, 8.3, 8.3 Hz), 6.94 (1H, s), 6.58 (1H, s), 3.48 (3H, s).

Example 5 <BR> <BR> <BR> <BR> <BR> 4- [Amino- [7- (3-cyano-phenyl)-5-nitro-benzofuran-2-yl]- (3-<BR> <BR> <BR> <BR> methyl-3H-imidazol-4-yl)-methyl]-benzonitrile In a similar manner to that described in Example 1-d) and 1-e), the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-nitro-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 3- [1, 3,2] dioxaborinan-2-yl-benzonitrile, followed by treatment with hydrogen chloride (4M solution in AcOEt) to give the titled compound as a colorless powder.

LCMS (condition B), m/z 475 [M+H] +, retention time 2.90 minutes lH-NMR (CD30D) d 8.60 (1H, d, J=2.0 Hz), 8. 46 (1H, d, J=2.0 Hz), 8.12 (2H, d, J=7.8 Hz), 7.80-7. 83 (3H, m), 7.64-7. 72 (4H, m), 6.99 (1H, s), 6.55 (1H, s), 3.41 (3H, s) Example 6 <BR> <BR> <BR> <BR> <BR> 4- [Amino- [7- (3-ethoxy-phenyl)-5-nitro-benzofuran-2-yl]- (3-<BR> <BR> <BR> <BR> methyl-3H-imidazol-4-yl)-methyl]-benzonitrile In a similar manner to that described in Example 1-d) and 1-e), the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-nitro-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3-ethoxy-phenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in AcOEt) to give the titled compound as a colorless powder.

LCMS (condition B), m/z 494 [M+H] +, retention time 3.10 minutesi lH-NMR (CD30D) d 8.51 (1H, d, J=2.3 Hz), 8.36 (1H, d, J=2.3 Hz), 7. 79 (2H, d, J=8.6 Hz), 7.62-7. 66 (3H, m), 7.30-7. 43 (3H, m) 6.96-7. 00 (1H, m), 6.92 (1H, s), 6.60 (1H, s), 4.00 (2H, q, J=6.9 Hz), 3.42 (3H, s) Example 7 <BR> <BR> <BR> <BR> <BR> <BR> <BR> 4- [Amino- [7- (3-chloro-phenyl)-5-nitro-benzofuran-2-yl]- (3-<BR> <BR> <BR> <BR> <BR> <BR> methyl-3H-imidazol-4-yl)-methyl]-benzonitrile In a similar manner to that described in Example 1-d) and 1-e), the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-nitro-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3-chloro-phenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in AcOEt) to give the titled compound as a colorless solid.

LCMS (condition B), m/z 484 [M+H] +, retention time 3.15 minutesi lH-NMR (CD30D) d 8.57 (1H, d, J=2.3 Hz), 8.40 (1H, d, J=2.3 Hz), 7.81 (2H, d, J=8.9 Hz), 7.70-7. 76 (2H, m), 7.62-7. 67 (3H, m), 7.43-7. 53 (2H, m), 7.00 (1H, s), 6.59 (1H, br s), 3.42 (3H, s) Example 8 <BR> <BR> <BR> <BR> <BR> <BR> <BR> 4- [Amino- [7- (4-fluoro-phenyl)-5-nitro-benzofuran-2-yl]- (3-<BR> <BR> <BR> <BR> <BR> <BR> methyl-3H-imidazol-4-yl)-methyl]-benzonitrile In a similar manner to that described in Example 1-d) and 1-e), the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-nitro-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (4-fluoro-phenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in AcOEt) to give the titled compound as a colorless powder.

LCMS (condition B), m/z 468 [M+H] +, retention time 3.06 minutes ; lH-NMR (CD30D) d 8.51 (1H, d, J=2.5 Hz), 8.36 (1H, d, J=2.5 Hz), 7.79-7. 84 (4H, m), 7.62-7. 70 (3H, m), 7.24 (2H, dd, J=8.79, 8,79 Hz), 6.87 (1H, s), 6.57 (1H, br s), 3.43 (3H, s) Example 9 <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> 4- [Amino- [7- (3, 5-difluoro-phenyl)-5-nitro-benzofuran-2-yl]-<BR> <BR> <BR> <BR> <BR> <BR> (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile In a similar manner to that described in Example 1-d) and 1-e), the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-nitro-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3, 5-difluoro-phenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in AcOEt) to give the titled compound as a colorless powder.

LCMS (condition B), m/z 486 [M+H] +, retention time 3.13 minutes ; 1H-NMR (CD30D) d 8.59 (1H, d, J=2.5 Hz), 8.43 (1H, d, J=2.5 Hz), 7.79 (2H, d, J=8.8 Hz), 7.63-7. 66 (3H, m), 7.40-7. 42 (2H, m), 7.04-7. 08 (1H, m), 6.97 (1H, s), 6.57 (1H, br s), 3.41 (3H, s) Example 10 <BR> <BR> <BR> <BR> <BR> <BR> <BR> <BR> 4- [Amino- [7- (3, 4-difluoro-phenyl)-5-nitro-benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile In a similar manner to that described in Example 1-d) and 1-e), the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-nitro-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3, 4-difluoro-phenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in AcOEt) to give the titled compound as a colorless powder.

LCMS (condition B), m/z 486 [M+H] +, retention time 3.09 minutes ; lH-NMR (CD30D) d 8.57 (1H, d, J=2.4 Hz), 8.39 (1H, d, J=2.4 Hz), 7.80 (2H, d, J=8.8 Hz), 7.73 (1H, ddd, J=2. 0, 7.3, 11.7 Hz), 7.62-7. 65 (4H, m), 7.41 (1H, ddd, J=8.79, 8.79, 10,7 Hz), 6.91 (1H, s), 6.57 (1H, br s), 3.43 (3H, s) Example 11 <BR> <BR> <BR> <BR> <BR> <BR> <BR> 4- [Amino- (3-methyl-3H-imidazol-4-yl)- (5-nitro-7-pyridin-3-<BR> <BR> <BR> <BR> <BR> <BR> yl-benzofuran-2-yl)-methyl]-benzonitrile In a similar manner to that described in Example 1-d) and 1-e),, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-nitro-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with pyridine-3-boronic acid, followed by treatment with hydrogen chloride (4M solution in AcOEt) to give the titled compound as a colorless powder.

LCMS (condition B), m/z 451 [M+H] +, retention time 2.27 minutes ; lH-NMR (CD30D) d 8.99 (1H, d, J=1. 6 Hz), 8.58-8. 61 (2H, m), 8.45 (1H, d, J=2.0 Hz), 8. 26-8. 29 (1H, m), 7.81 (2H, d, J=6.4 Hz), 7.65 (2H, d, J=6.4 Hz), 7.57-7. 60 (2H, m), 6.92 (1H, s), 6.57 (1H, s), 3.46 (3H, s) Example 12 <BR> <BR> <BR> <BR> <BR> <BR> <BR> 4- [Amino- (3-methyl-3H-imidazol-4-yl)- (5-nitro-benzofuran-2-<BR> <BR> <BR> <BR> <BR> <BR> yl)-methyl]-benzonitrile In a similar manner to that described in Example 1-a) and 1-e), 2-hydroxy-5-nitro-benzaldehyde was used instead of 3-bromo-2-hydroxy-5-nitro-benzaldehydethe. Suzuki coupling reaction of 2-methyl-propane-2-sulfinic acid [ (5- nitro-benzofuran-2-yl)- (4-cyano-phenyl)- (3-methyl-3H- imidazol-4-yl)-methyl]-amide with phenylboronic acid,

followed by treatment with hydrogen chloride (4M solution in AcOEt) to give the titled compound as a colorless powder.

EI-MS: m/z 373 [M] + ; 1H-NMR (CDC13) d 8.37 (1H, br-s), 8.18 (1H, d, J=9.1 Hz), 7.65 (2H, d, J=8.9 Hz), 7.47-7. 52 (3H, m), 7.39 (1H, s), 6.55 (1H, s), 6.38 (1H, s), 3.37 (3H, s), 2.40 (2H, brs) Preparation of 2- [amino- (4-cyano-phenyl)- (3-methyl-3H- imidazol-4-yl) -methyl] -benzofuran-5-carbonitrile compounds Example 13 Preparation of 2- [amino- (4-cyano-phenyl)- (3-methyl-3H- imidazol-4-yl)-methyl]-7-phenyl-benzofuran-5-carbonitrile 13-a) Preparation of 3-bromo-5-formyl-4-hydroxy- benzonitrile, 11 To a solution of 3-formyl-4-hydroxy-benzonitrile (6.65 g, 45.2 mmol) [Suzuki, Y. et al. Chem. Pharm. Bull. 1983, 31, 1751-1753] in EtOH (200 ml) was added dropwise bromine (7 ml, 136.6 mmol) over 15 minutes at room temperature. The resulting mixture was stirred at room temperature for 30 minutes. The solvent was evaporated to give 3-bromo-5- formyl-4-hydroxy-benzonitrile (crude, 10.2 g) which was used for the next step without further purification.

EI-MS m/z 225 (M+), 227 (M+2) ; 1H-NMR (CDCl3) 612. 08 (1H, s), 9.90 (1H, s), 8.05 (1H, d, J=2 Hz), 7.91 (1H, d, J=2 Hz) 13-b) Preparation of 7-bromo-2- (4-cyano-benzoyl)- benzofuran-5-carbonitrile The crude of 3-bromo-5-formyl-4-hydroxy-benzonitrile (10.2 g) was added to a solution of 4-cyanophenacyl bromide

(11.7 g, 49. 6 mmol) and K2CO3 (7.5 g, 54. 3 mmol) in CH3CN (250 ml). The mixture was stirred for 30 minutes under reflux, and then the solvent was evaporated in vacuo. The residue was washed with AcOEt and H2O, then MeOH to give 13.8 g of 7-bromo-2- (4-cyano-benzoyl)-benzofuran-5- carbonitrile as a light brown powder.

EI-MS m/z 350 (M+), 352 (M+2); lH-NMR (CDC13) 68. 26 (2H, dt, Je=8. 5 Hz, J2=2 Hz), 8. 09 (1H, d, J=1 Hz), 7. 96 (1H, d, J=1 Hz), 7.89 (2H, dt, J1=8. 5 Hz, J2=2 Hz), 7.76 (1H, s) 13-c) Preparation of 2-methyl-propane-2-sulfinic acid (7- bromo-5-cyano-benzofuran-2-yl)- (4-cyano-phenyl)- methyleneamide To a solution 7-bromo-2- (4-cyano-benzoyl)-benzofuran-5- carbonitrile (13.5 g, 38.5 mmol) in dry THF (500 ml) was added Ti (OEt) 4 (23.8 ml, 115.8 mmol) at room temperature.

To the mixture was then added 2-methyl-2-propane sulfinamide (5.13 g, 42.3 mmol), and the mixture was stirred for 8 hours under reflux. After cooling to room temperature, the mixture was poured into an equal volume of brine while being stirred rapidly. The resulting suspension was filtered through a pad of Celite, and the filter cake was washed with AcOEt. The filtrate was extracted with AcOEt (x 2). The organic layers were combined, washed with brine, dried over Na2SO4 and evaporated in vacuo. The residue was purified by column chromatography over silica gel (eluent; CH2Cl2-> 5 % MeOH in DCM), yielding 7.5 g of 2-methyl-propane-2-sulfinic acid (7-bromo-5-cyano- benzofuran-2-yl)- (4-cyano-phenyl)-methyleneamide as a yellow amorphous powder.

LCMS (condition B) m/z 454 [M+H] +, retention time 3.84 minutes ; 1H-NMR (CDC13) d 7.93 (1H, br-s), 7. 88-7. 87 (1H, m), 7.78 (2H, d, J=8.5 Hz), 7. 65-7. 62 (2H, m), 7.08 (1H, br-s).

13-d) Preparation of 2-methyl-propane-2-sulfinic acid [ (7- <BR> <BR> <BR> bromo-5-cyano-benzofuran-2-yl)- (4-cyano-phenyl)- (3-methyl- 3H-imidazol-4-yl)-methyl]-amide, n-Butyllithium in hexane (1.57 M, 11.6 ml, 18.2 mmol) was added to a solution of N-methylimidazole (1.45 ml, 18.2 mmol) in THF (30 ml) at-78 °C. After stirring for 30 minutes, triethylsilyl chloride (3.1 ml, 18.5 mmol) was added to the reaction mixture. The mixture was allowed to warm to room temperature then cooled to-78 °C. n- Butyllithium in hexane (1.57 M, 11.6 ml, 18.2 mmol) was added again. The reaction mixture was allowed to warm to- 20 °C and then cooled to-78 °C. The resulting mixture was added to a solution of 2-methyl-propane-2-sulfinic acid (7- bromo-5-cyano-benzofuran-2-yl)- (4-cyano-phenyl)- methyleneamide (5.5 g, 12.1 mmol) in THF (115 ml) at-78 °C.

After stirring for 15 minutes, saturated NH2Cl was added.

The reaction mixture was extracted with AcOEt (x 2). The organic layers were combined, washed with brine, dried over Na2SO4 and evaporated in vacuo. The residue was purified by column chromatography over silica gel (eluent; 2 % MeOH in CH2DCM-> 5 % MeOH in DCM), yielding 3.62 g of 2-methyl- propane-2-sulfinic acid [ (7-bromo-5-cyano-benzofuran-2-yl)- <BR> <BR> <BR> (4-cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide as a colorless solid.

LCMS (condition B) m/z 536 [M+H] +, retention time 2.78 minutes ; 1H-NMR (CDC13) d 7.85 (1H, d, J=1.5 Hz), 7.74 (2H, dt, Je=8. 5 Hz, J2=2 Hz), 7. 73 (1H, s), 7.63 (2H, dt, Jl=8. 5 Hz, J2=2 Hz), 7.54 (1H, s), 7.19 (1H, s), 6.94 (1H, d, J=1.5 Hz), 4.65 (1H, s), 3.15 (3H, s), 1.35 (9H, s).

13-e) Preparation of 2-methyl-propane-2-sulfinic acid [ (4- <BR> <BR> <BR> cyano-phenyl)- (5-cyano-7-phenyl-benzofuran-2-yl)- (3-methyl-<BR> <BR> <BR> <BR> 3H-imidazol-4-yl)-methyl]-amide

A mixture of 2-methyl-propane-2-sulfinic acid [ (7- <BR> <BR> <BR> <BR> bromo-5-cyano-benzofuran-2-yl.)- (4-cyano-phenyl)- (3-methyl-.

3H-imidazol-4-yl) -methyl] -amide (800 mg, 1.49 mmol), 2- phenyl-1, 3,2-dioxabororinane (0.67 ml, 4.47 mmol), tetrakis (triphenyl-phosphine)-palladium (0) (345 mg, 0.30 mmol) and tripotassium phosphate (1.27 mg, 5.98 mmo) in DMF (24 ml) was heated at 100 °C for 1 hour. The reaction mixture was diluted with AcOEt and then palladium was filtered off. The filtrate was extracted with AcOEt. The organic layers were combined, washed with brine, dried over Na2SO4 and evaporated in vacuo. The residue was purified by column chromatography over silica gel (eluent; 1 % MeOH in DCM-> 10 % MeOH in DCM), yielding 714 mg of 2-methyl- propane-2-sulfinic acid [ (4-cyano-phenyl)- (5-cyano-7- phenyl-benzofuran-2-yl)- (3-methyl-3H-imidazol-4-yl)- methyl] -amide as a colourless powder.

LCMS (condition B) m/z 534 [M+H] +, retention time 3.01 minutes ; 1H-NMR (CDC13) d 7.86 (1H, d, J=1.5 Hz), 7.71 (2H, d, J=8.5 Hz), 7.67 (1H, d, J=1.5 Hz), 7.57 (2H, d, J=8.5 Hz), 7. 58-7. 53 (3H, m), 7. 46-7. 42 (3H, m), 7.20 (1H, s), 6.87 (1H, br-s), 4.68 (1H, s), 3.13 (3H, s), 1.30 (9H, s).

13-f) Preparation of 2- [Amino- (4-cyano-phenyl)- (3-methyl- 3H-imidazol-4-yl)'-methyl]-7-phenyl-benzofuran-5- carbonitrile To a mixture of 2-methyl-propane-2-sulfinic acid [ (4- <BR> <BR> <BR> <BR> cyano-phenyl)- (5-cyano-7-phenyl-benzofuran-2-yl)- (3-methyl- 3H-imidazol-4-yl) -methyl] -amide (687 mg, 1.28 mmol) in THF (10 ml) was added 4M HC1 solution in dioxane (5 ml) at room temperature. After stirring at room temperature for 1 hour, the reaction mixture was quenched with saturated NaHCO3 at 0 °C and extracted with AcOEt (x 2). The organic layers were combined and washed with brine, dried over Na2SO4 and evaporated in vacuo. The residue was purified by MPLC [CPO- HS-221-Si-10 KUSANO, 22X300mm] (eluent; 2. 5% MeOH in DCM),

yielding 477 mg of 2- [amino- (4-cyano-phenyl)- (3-methyl-3H- imidazol-4-yl)-methyl]-7-phenyl-benzofuran-5-carbonitrile as a colorless powder.

LCMS (condition B) m/z 430 [M+H] +, retention time 2.90 minutes ; 1H-NMR (CDC13) d 7.81 (1H, d, J=1.5 Hz), 7. 73-7. 68 (4H, m), 7.67 (1H, d, J=1.5 Hz), 7.55 (2H, dt, J1=8. 5 Hz, J2=2 Hz), 7. 51-7. 43 (4H, m), 6.66 (1H, d, J=1 Hz), 6.44 (1H, s), 3.39 (3H, s), 2.46 (2H, br-s).

Example 14 Preparation of 2- [amino- (4-cyano-phenyl)- (3-methyl-3H- imidazol-4-yl)-methyl]-7- (3-methoxy-phenyl)-benzofuran-5- carbonitrile In a similar manner to that described in Example 13-e and 13-f, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-cyano-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3-methoxyphenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in 1,4- dioxane) to give the titled compound as a colorless powder.

LCMS (condition B) m/z 460 [M+H] +, retention time 2.94 minutes ; 1H-NMR (CDC13) d 7.81 (1H, d, J=1.5 Hz), 7.73 (1H, d, J=1.5 Hz), 7.72 (2H, dt, J1=8. 5 Hz, J2=2 Hz), 7.56 (2H, dt, J1=8. 5 Hz, J2=2 Hz), 7.44 (1H, br-d, J=1 Hz), 7.40 (1H, t, J=8 Hz), 7. 26 (1H, dt, Jl=8 Hz, J2=2 Hz), 7.21 (1H, br-t, J=2 Hz), 6.98 (1H, ddd, J1=8 Hz, J2=2 Hz, J3=1 Hz), 6.67 (1H, d, J=1 Hz), 6.46 (1H, s), 3.83 (3H, s), 3.39 (3H, s), 2.45 (2H, br-s).

Example 15 Preparation of 2- [amino- (4-cyano-phenyl)- (3-methyl-3H- imidazol-4-yl)-methyl]-7-benzo [1, 3] dioxol-5-yl-benzofuran- 5-carbonitrile

In a similar manner to that described in Example 13-e and 13-f, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-cyano-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 5- [1, 3,2] dioxaborinan-2-yl-benzo [1, 3] dioxane, followed by treatment with hydrogen chloride (4M solution in 1,4- dioxane) to give the titled compound as a colorless powder.

LCMS (condition B) m/z 474 [M+H] +, retention time 2.87 minutes ; lH-NMR (CDCl3) d 7.77 (1H, d, J=1.5 Hz), 7.73 (2H, dt, Jl=9 Hz, J2=2 Hz), 7.65 (1H, d, J=1.5 Hz), 7. 56 (2H, dt, J1=9 Hz, J2=2 Hz), 7.44 (1H, br-d, J=1 Hz), 7.17 (1H, dd, J1=8 Hz, J2=2 Hz), 7.13 (1H, d, J=2 Hz), 6.92 (1H, d, J=8 Hz), 6.63 (1H, d, J=1 Hz), 6.47 (1H, s), 6.05 (1H, d, J=1 Hz), 6.04 (1H, d, J=1 Hz), 3.39 (3H, s), 2.47 (2H, br-s).

Example 16 Preparation of 2-[Amino-(4-cyano-phenyl)-(3-methyl-3H- imidazol-4-yl)-methyl]-7- (3-fluoro-phenyl)-benzofuran-5- carbonitrile In a similar manner to that described in Example 13-e and 13-f, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-cyano-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3-fluorophenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in 1,4- dioxane) to give the titled compound as a colorless powder.

LCMS (condition B) m/z 448 [M+H] +, retention time 2.92 minutes; 1H-NMR (CDCl3) d 7.85 (1H, d, J=1.5 Hz), 7.73 (2H, d, J=8.5 Hz), 7.72 (1H, d, J=1.5 Hz), 7. 56 (2H, d, J=8.5 Hz), 7. 48-7. 45 (3H, m), 7. 40-7. 32 (1H, m), 7. 20-7. 11 (1H, m), 6.62 (1H, d, J=1 Hz), 6.50 (1H, s), 3.39 (3H, s), 2.48 (2H, br-s).

Example 17

Preparation of 2- [amino- (4-cyano-phenyl)- (3-methyl-3H- imidazol-4-yl)-methyl]-7- (3-cyano-phenyl)-benzofuran-5- carbonitrile In a similar manner to that described in Example 13-e and 13-f, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-cyano-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3-cyanophenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in 1,4- dioxane) to give the titled compound as a colorless powder.

LCMS (condition B) m/z 455 [M+H] +, retention time 2.76 minutes ; lH-NMR (CDCl3) d 7.98 (1H, br-s), 7.92 (1H, dt, J1=8 Hz, J2=1 Hz), 7. 89 (1H, d, J=1 Hz), 7. 76 (2H, d, J=8 Hz), 7.74 (1H, d, J=8 Hz), 7.72 (1H, d, J=1 Hz), 7.63 (1H, t, J=8 Hz), 7. 56 (2H, d, J=8 Hz), 7. 46 (1H, s), 6.58 (1H, d, J=1 Hz), 6.54 (1H, s), 3.40 (3H,'s), 2.48 (2H, br-s) Example 18 Preparation of 2- [amino- (4-cyano-phenyl)- (3-methyl-3H- <BR> <BR> <BR> <BR> imidazol-4-yl)-methyl]-7- (3-chloro-phenyl)-benzofuran-5-<BR> <BR> <BR> <BR> <BR> carbonitril In a similar manner to that described in Example 13-e and 13-f, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-cyano-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3-chlorophenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in 1,4- dioxane) to give the titled compound as a colorless powder.

LCMS (condition B) m/z 464 [M+H] +, retention time 3.05 minutes ; lH-MMR (CDC13) d 7.85 (1H, d, J=1.5 Hz), 7.74 (2H, dt, J1=8. 5 Hz, J2=2 Hz), 7.71 (1H, d, J=1.5 Hz), 7.64 (1H, m), 7.58 (1H, m), 7.56 (2H, d, J=8.5 Hz), 7.43 (2H, m),

7.42 (1H, d, J=1 Hz), 6.63 (1H, d, J=1 Hz), 6.56 (1H, s), 3.38 (3H, s), 2.47 (2H, br-s).

Example 19 Preparation of 2- [Amino- (4-cyano-phenyl)- (3-methyl-3H- imidazol-4-yl)-methyl]-7-pyridin-3-yl-benzofuran-5- carbonitrile In a similar manner to that described in Example 13-e and 13-f, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-cyano-benzofuran-2-yl)- (4- cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 3- [1, 3,2] dioxaborinan-2-yl-pyridine, followed by treatment with hydrogen chloride (4M solution in 1,4- dioxane) to give the titled compound as a colorless powder.

LCMS (condition B), m/z 431 [M+H] +, retention time 2.16 minutes ; 1H NMR (CD30D) d 8.96 (1H, d, J=1.6 Hz), 8.59 (1H, dd, J=1.6, 4.8 Hz), 8.25 (1H, ddd, J=2.0, 2.0, 8.0 Hz), 8. 08 (1H, d, J=1.4 Hz), 7.94 (1H, d, J=1.4 Hz), 7.81 (2H, d, J=8.8 Hz), 7.55-7. 65 (4H, m), 6.84 (1H, s), 6.55 (1H, s), 3.43 (3H, s).

Preparation of 4- [amino- (5-pyridin-3-yl-benzofuran-2-yl)- (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile compounds Example 20 Preparation of 4- [amino- [7- (3-fluoro-phenyl)-5-pyridin-3- yl-benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile 20-a) Preparation of 3-bromo-2-hydroxy-5-pyridine-3-yl- benzaldehyde To a solution of 2-hydroxy-5-pyridine-3-yl-benzaldehyde (20.0 g, 100 mmol) in DCM/MeOH (500/200 ml) was added benzyltrimethylammonium tribrommide (39.1 g, 100 mmol) at

room temperature. After stirring at room temperature for 2h, the mixture was quenched with water. The mixture was extracted with DCM (x 2). The organic layers were combined, dried over MgS04, filtered and evaporated in vacuo to give 25.0 g of 3-bromo-2-hydroxy-5-pyridine-3-yl-benzaldehyde as a light yellow solid (crude).

LCMS (condition B) m/z 278 [M+H] +, retention time 2.54 minutes ; 1H-NMR (CDC13) d 11.65 (s, 1H), 9.96 (s, 1H), 8.83 (d, J= 1.7 Hz, 1H), 8.64 (dd, J= 1.7, 4.8 Hz, 1H), 8.03 (d, J= 2.3 Hz, 1H), 7.86 (ddd, J= 1.7, 2.3, 7.9 Hz, 1H), 7.76 (d, J= 2.3 Hz, 1H), 7.41 (dd, J= 4.8, 7.9 Hz, 1H).

20-b) Preparation of 4- (7-bromo-5-pyridin-3-yl-benzofuran- 2-carbonyl)-benzonitrile 3-Bromo-2-hydroxy-5-pyridine-3-yl-benzaldehyde (12.0 g, 43.1 mmol) was added to a solution of 4-cyanophenacyl bromide (10.6 g, 47.4 mmol) and K2CO3 (59.6 g, 431 mmol) in CH3CN (200 ml). The mixture was stirred at 60 °C for 1 hour under argon, the solvent was evaporated in vacuo. The residue was washed with HzO and MeOH to give 6.5 g of 4- (7- bromo-5-pyridin-3-yl-benzofuran-2-carbonyl) -benzonitrile as a light brown powder.

LCMS (condition B) m/z 403 [M+H] +, retention time 2.90 minutes ; 1H NMR (DMSO) d 8.96 (d, J= 2.3 Hz, 1H), 8.62 (dd, J= 1.6, 4.6 Hz, 1H), 8.10-8. 23 (m, 7H), 7.97 (s, 1H), 7.53 (dd, J= 4.6, 7.9 Hz, 1H).

20-c) Preparation of 2-methyl-propane-2-sulfinic acid (7- bromo-5-pyridin-3-yl-benzofuran-2-yl)- (4-cyano-phenyl)- methyleneamide, To a solution of 4- (7-bromo-5-pyridin-3-yl-benzofuran- 2-carbonyl) -benzonitrile (6.5 g, 16.4 mmol) in THF (200 ml) was added Ti (OEt) 4 (11.1 ml, 54.1 mmol) and 2-methyl-2- propane sulfinamide (2.2 g, 18.0 mmol) at room temperature.

The mixture was stirred for 4 hours under reflux. After the reaction mixture was cooled to room temperature, brine was added and stirred rapidly. The resulting suspension was filtered through a pad of Celite, and the filter cake was washed with EtOAc. The filtrate was partitioned, the aqueous layer was extracted with EtOAc (x 2). The organic layers were combined, dried over MgSO4 and evaporated in vacuo. The residue was purified by column chromatography over silica gel (eluent; CH2Cl2-5 % MeOH in DCM), yielding 3.8 g of 2-methyl-propane-2-sulfinic acid (7-bromo-5- pyridin-3-yl-benzofuran-2-yl)- (4-cyano-phenyl)- methyleneamide as an orange amorphous powder.

LCMS (condition B) m/z 506 [M+], 508 [M+2], retention time 3. 00 minutes ; 1H NMR (CDC13) d 8. 84 (d, J= 1.7 Hz, 1H), 8.64 (dd, J= 1. 7,4. 8 Hz, 1H), 7.87 (ddd, J= 1.7, 2.6, 7.9 Hz, 1H), 7.74-7. 83 (m, 7H), 7.41 (dd, J= 4.8, 7.9 Hz, 1H), 1.40 (s, 9H).

20-d) Preparation of 2-Methyl-propane-2-sulfinic acid [ (7- <BR> <BR> <BR> bromo-5-pyridin-3-yl-benzofuran-2-yl)- (4-cyano-phenyl)- (3- methyl-3H-imidazol-4-yl)-methyl]-amide, 21 n-Butyllithium in hexane (1.57 M, 7.4 ml, 11.6 mmol) was added to a solution of N-methylimidazole (0.84 ml, 10.5 mmol) in THF (18 ml) at-78 °C. After stirring for 15 minutes, triethylsilyl chloride (1.9 ml, 11.6 mmol) was added to the reaction mixture. The mixture was allowed to warm to room temperature then cooled to-78 °C again. n- Butyllithium in hexane (1.57 M, 6.7 ml, 10.5 mmol) was added. The reaction mixture was allowed to warm to-20 °C and then cooled to-78 °C. The resulting mixture was added to a solution of 2-methyl-propane-2-sulfinic acid (7-bromo- 5-pyridin-3-yl-benzofuran-2-yl)- (4-cyano-phenyl)- methyleneamide (3.8 g, 7.50 mmol) in THF (70 ml) at-78 °C.

After stirring for 15 minutes, saturated NH3Cl was added.

The reaction mixture was extracted with EtOAc (x 2). The

organic layers were combined, dried over MgS04 and evaporated in vacuo. The residue was purified by column chromatography over silica gel (eluent; DCM-> 5 % MeOH in DCM), yielding 3.8 g of 2-methyl-propane-2-sulfinic acid <BR> <BR> <BR> [ (7-bromo-5-pyridin-3-yl-benzofuran-2-yl)- (4-cyano-phenyl)- (3-methyl-3H-imidazol-4-yl) -methyl] -amide as a light yellow amorphous powder.

LCMS (condition B) m/z 588 [M+], 590 [M+2], retention time 2.39 minutes ; 1H-NMR (CDC13) d 8.81 (d, J= 2.3 Hz, 1H), 8.62 (dd, J= 1.7, 4.6 Hz, 1H), 7.84 (ddd, J= 1.7, 2.3, 7.9 Hz, 1H), 7.72-7. 76 (m, 2H), 7.64-7. 67 (m, 4H), 7.54 (s, 1H), 7.39 (dd, J= 4.6, 7.9 Hz, 1H), 7.03 (s, 1H), 6.97 (s, 1H), 4.63 (s, 1H), 3.22 (s, 3H), 1.35 (s, 9H).

20-e) Preparation of 2-methyl-propane-2-sulfinic acid [ (4- <BR> <BR> <BR> cyano-phenyl)- [7- (3-fluoro-phenyl)-5-pyridin-3-yl-<BR> <BR> <BR> <BR> <BR> benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]-amide A mixture of 2-methyl-propane-2-sulfinic acid [ (7- <BR> <BR> <BR> bromo-5-pyridin-3-yl-benzofuran-2-yl)- (4-cyano-phenyl)- (3- methyl-3H-imidazol-4-yl) -methyl] -amide (1.5 g, 2.55 mmol), 2- (3-fluoro-phenyl)- [1, 3,2] dioxaborinane (1.3 g, 7.65 mmol), tetrakis (triphenylphosphine)-palladium (0) (589 mg, 0.51 mmol) and tripotassium phosphate (1.6 g, 7.65 mmo) in DMF (20 ml) was heated at 100 °C for 2 hour. The reaction mixture was diluted with EtOAc and then palladium was filtered off. The filtrate was partitioned between EtOAc and H20. The water layer was extracted with EtOAc (x 2).

The organic layers were combined, dried over MgS04 and evaporated in vacuo. The residue was purified by column chromatography over silica gel (eluent; DCM-> 5 % MeOH in DCM), yielding 1.1 g of 2-methyl-propane-2-sulfinic acid <BR> <BR> <BR> [ (4-cyano-phenyl)- [7- (3-fluoro-phenyl)-5-pyridin-3-yl-<BR> <BR> <BR> <BR> <BR> benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]-amide as a light yellow powder.

LCMS (condition B) m/z 604 [M+H] +, retention time 2.54 minutes ; 1H-NMR (CDC13) d 8.88 (d, J= 2.0 Hz, 1H), 8.63 (dd, J=l 1. 7,4. 9 Hz, 1H), 7.92 (dt, J= 2.0, 8.2 Hz, 1H), 7.37- 7.25 (m, 11H), 7.17 (s, 1H), 7.10 (ddt, J= 1.3, 2.5, 8.2 Hz, 1H), 6.93 (s, 1H), 4.67 (s, 1H), 3.18 (s, 3H), 1.31 (s, 9H).

20-f) Preparation of 4- [amino- [7- (3-fluoro-phenyl)-5- <BR> <BR> <BR> pyridin-3-yl-benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)- methyl]-benzonitrile To a mixture of 2-methyl-propane-2-sulfinic acid [ (4- <BR> <BR> <BR> cyano-phenyl)- [7- (3-fluoro-phenyl)-5-pyridin-3-yl-<BR> <BR> <BR> <BR> <BR> benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]-amide (1.1 g, 1.82 mmol) in THF (15 ml) was added 4M HC1 solution in EtOAc (7 ml) at room temperature. The mixture was stirred at room temperature for 15 hour. The reaction mixture was quenched with saturated NaHCO3 and extracted with EtOAc (x 2). The organic layers were combined, dried over MgS04 and evaporated in vacuo. The residue was purified by column chromatography over silica gel (eluent; 3 % MeOH in DCM), yielding 810 mg of 4- [amino- [7- (3-fluoro- phenyl)-5-pyridin-3-yl-benzofuran-2-yl]- (3-methyl-3H- imidazol-4-yl)-methyl]-benzonitrile as a colorless powder.

LCMS (condition B) m/z 500 [M+H] +, retention time 2.46 minutes ; 1H-NMR (CDC13) d 8.89 (dd, J= 1.0, 2.6 Hz, 1H), 8.62 (dd, J= 1.7, 4.9 Hz, 1H), 7.92 (dt, J= 2.2, 8.6 Hz, 1H), 7.69-7. 74 (m, 3H), 7.54-7. 64 (m, 4H), 7.38-7. 50 (m, 4H), 7.11 (ddt, J= 1.0, 2.6, 8.6 Hz, 1H), 6.68 (d, J= 1.0 Hz, 1H), 6.49 (s, 1H), 3.42 (s, 3H), 2.49 (brs, 1H).

Example 21 Preparation of 4- [amino- (3-methyl-3H-imidazol-4-yl)- (7- <BR> <BR> <BR> phenyl-5-pyridin-3-yl-benzofuran-2-yl)-methyl]-benzonitrile In a similar manner to that described in Example 20-e and 20-f, the Suzuki coupling reaction of 2-methyl-propane-

2-sulfinic acid [ (7-bromo-5-pyridin-3-yl-benzofuran-2-yl)- <BR> <BR> (4-cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2-phenyl-1, 3,2-dioxabororinane, followed by treatment with hydrogen chloride (4M solution in EtOAc) to give the titled compound as a colorless powder.

LCMS (condition B) m/z 482 [M+H] +, retention time 2.63 minutes ; 1H-NMR (CD30D) d 8.87 (d, J= 2.3 Hz, 1H), 8.52 (d, J= 4.8 Hz, 1H), 8.17 (d, J= 8.0 Hz, 1H), 7.73-7. 84 (m, 6H), 7.62-7. 65 (m, 3H), 7.53 (dd, J= 4.8, 8.0 Hz, 1H) 7.35-7. 48 (m, 3H), 6.76 (s, 1H), 6.58 (s, 1H), 3.42 (s, 3H).

Example 22 Preparation of 4- [amino- [7- (3-methoxy-phenyl)-5-pyridin-3- yl-benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile In a similar manner to that described in Example 20-e and 20-f, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-pyridin-3-yl-benzofuran-2-yl)- <BR> <BR> <BR> (4-cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3-methoxy-phenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in EtOAc) to give the titled compound as a colorless powder.

LCMS (condition B) m/z 512 [M+H] +, retention time 2.43 minutes ; 1H-NMR (CDC13) d 8.89 (d, J= 2.0 Hz, 1H), 8.62 (dd, J= 1.7, 4.9 Hz, 1H), 7.92 (dt, J= 2.0, 8.6 Hz, 1H), 7.72 (dd, J= 2.0, 6.6 Hz, 2H), 7.66 (dd, J= 2.0, 4.5 Hz, 2H), 7.58 (dd, J= 2.0, 6.6 Hz, 2H), 7.31-7. 44 (m, 5H), 6.96 (ddd, J= 1.7, 2.6, 7.6 Hz, 1H), 6.72 (d, J= 1.0 Hz, 1H), 6.45 (s, 1H), 3.83 (s, 3H), 3.42 (s, 3H), 2.45 (brs, 2H).

Example 23

Preparation of 4- [amino- (7-benzo [1, 3] dioxol-5-yl-5-pyridin- <BR> <BR> <BR> <BR> 3-yl-benzofuran-2-yl)- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile In a similar manner to that described in Example 20-e and 20-f, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-pyridin-3-yl-benzofuran-2-yl)- <BR> <BR> <BR> <BR> (4-cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 5- [1, 3,2] dioxa-borinan-2-yl-benzo [1, 3] dioxole, followed by treatment with hydrogen chloride (4M solution in EtOAc) to give the titled compound as a colorless powder.

LCMS (condition B) m/z 526 [M+H] +, retention time 2.41 minutes 1H-NMR (CD30D) d 8.85 (s, 1H), 8.52 (d, J= 4.9 Hz, 1H), 8.16 (d, J= 6.4 Hz, 1H), 7.78-7. 80 (m, 3H), 7.63-7. 66 (m, 4H) 7.53 (dd, J= 4.9, 7.8 Hz, 1H), 7.31 (d, J= 7.8 Hz, 1H), 7.25 (s, 1H), 6.91 (d, J= 7.8 Hz, 1H), 6.77 (s, 1H), 6.57 (s, 1H), 6.00 (s, 2H), 3.48 (s, 3H).

Example 24 Preparation of 4- [amino- [7- (3-cyano-phenyl)-5-pyridin-3-yl- benzofuran-2-yl]-(3-methyl-3H-imidazol-4-yl ?-methyl]- benzonitrile In a similar manner to that described in Example 20-e and 20-f, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-pyridin-3-yl-benzofuran-2-yl)- <BR> <BR> <BR> <BR> (4-cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3-cyanophenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in EtOAc) to give the titled compound as a colorless powder.

LCMS (condition B) m/z 507 [M+H] +, retention time 2.43 minutes ; 1H-NMR (CD30D) d 8.91 (d, J= 2.0 Hz, 1H), 8.54 (dd, J= 1.5, 4.9 Hz, 1H), 8.21 (dt, J= 1.5, 7.8 Hz, 1H), 8.15- 8.17 (m, 2H), 7.95 (d, J= 2.0 Hz, 1H), 7.82-7. 84 (m, 3H) 7.76 (dt, J= 1.5, 7.1 Hz, 1H), 7.64-7. 69 (m, 4H), 7.55 (dd,

J= 4.9, 8.8 Hz, 1H), 6.87 (s, 1H), 6.55 (s, 1H), 3.43 (s, 3H).

Example 25 Preparation of 4- [amino- [7- (3-chloro-phenyl)-5-pyridin-3- yl-benzofuran-2-yl]- (3-methyl-3H-imidazol-4-yl)-methyl]- benzonitrile In a similar manner to that described in Example 20-e and 20-f, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-pyridin-3-yl-benzofuran-2-yl)- <BR> <BR> <BR> (4-cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 2- (3-chlorophenyl)- [1, 3,2] dioxaborinane, followed by treatment with hydrogen chloride (4M solution in EtOAc) to give the titled compound as a colorless powder.

LCMS (condition A) m/z 516 [M+H] +, retention time 2.59 minutes ; 1H-NMR (CDC13) d 8.87 (d, J= 2.6 Hz, 1H), 8.63 (dd, J= 1.7, 5.0 Hz, 1H), 7.92 (ddd, J= 1.7, 2.6, 8.9 Hz, 1H), 7.69-7. 75 (m, 4H), 7.63-7. 67 (m, 1H), 7.57-7. 62 (m, 3H), 7.37-7. 45 (m, 4H), 6.69 (d, J= 1. 0 Hz, 1H), 6.53 (s, 1H), 3.41 (s, 3H), 2.48 (brs, 1H).

Example 26 Preparation of 4- [amino- (5, 7-di-pyridin-3-yl-benzofuran-2- yl)- (3-methyl-3H-imidazol-4-yl)-methyl]-benzonitrile In a similar manner to that described in Example 20-e and 20-f, the Suzuki coupling reaction of 2-methyl-propane- 2-sulfinic acid [ (7-bromo-5-pyridin-3-yl-benzofuran-2-yl)- <BR> <BR> <BR> (4-cyano-phenyl)- (3-methyl-3H-imidazol-4-yl)-methyl]-amide with 3- [1, 3,2] dioxaborinan-2-yl-pyridine, followed by treatment with hydrogen chloride (4M solution in EtOAc) to give the titled compound as a colorless powder.

LCMS (condition B) m/z 483 [M+H] +, retention time 1.93 minutes ; 1H-NMR (CD30D) d 9.04 (d, J= 1.5 Hz, 1H), 8.91 (d,

J= 1.5 Hz, 1H), 8.54-8. 57 (m, 2H), 8.32 (ddd, J= 1.5, 2.4, 8.8 Hz, 1H), 8.21 (ddd, J= 1.5, 2.4, 7.3 Hz, 1H), 7.95 (d, J= 2.0 Hz, 1H), 7.85 (d, J= 1.5 Hz, 1H), 7.82 (d, J= 8.8 Hz, 2H), 7.65 (d, J= 8.8 Hz, 2H), 7.64-7. 67 (m, 1H), 7.54-7. 59 (m, 2H), 6. 81 (s, 1H), 6. 56 (s, 1H), 3. 44 (s, 3H).

Example 27 In vitro assay for inhibition of farnesyl-protein transferase and geranylgeranyl-protein transferase Human farnesyl-protein transferase alpha and beta subunits were coexpressed in Escherichia coli BL21 CodonPlus (DE3) RIL (Stratagene), essentially as described method (Methods Enzymol. 1995 Vol. 250 pp3-12 Bacterial expression and purification of human protein prenyltransferases using epitope-tagged, translationally coupled systems. Omer, Charles A. Diehl, Ronald E. and Kral, Astrid M. ) except that pT7-7 (Comb Chem High Throughput Screen. 1999 Vol. 5 pp279-87 Application of homogeneous time-resolved fluorescence (HTRFTM) to monitor poly-ubiquitination of wild-type p53 Yabuki, N. , Watanabe, S. , Kudoh, T. , Nihira, Si. , and Miyamato, C. ) was used.

Cultured cells were disrupted by sonication in 5mM sodium phosphate buffer (pH7.2), containing 75mM NaCl, 50uM ZnCl2, 5mM dithiothreitol, 0.5mM 4- (amidinophenyl) methanesulphonyl fluoride, and protease inhibitor cocktail tablet (Complete Mini EDTA-free, Roche Diagnostics). The crude extracts are centrifuged at 40,000 x g for 30 min at 4 °C. From the supernatant, human farnesyl-protein transferase was purified by three column chromatography, DEAE Sephacel (Amersham Pharmacia Biotech), HiTrapQ column (Amersham Pharmacia Biotech), and Phenyl Sepharose High Performance column (Amersham Pharmacia Biotech).

Human geranylgeranyl-protein transferase alpha and beta subunits were coexpressed in E. coli, BL21

CodonPlus (DE3) RIL, using pGEX4T-1 (Amersham Pharmacia Biotech). Cultured cells were disrupted by sonication in phosphate-buffered saline containing 1% of Triton X-100.

The crude extracts are centrifuged at 40,000 x g for 30 min at 4 °C. From the supernatant, geranylgeranyl-protein transferase was purified by Glutathione Sepharose 4FF (Amersham Pharmacia Biotech) and HiTrapQ (Amersham Pharmacia Biotech) column chromatography. The geranylgeranyl-protein transferase containing fractions were pooled and dialyzed against 50mM Tris-Cl pH7.5 containing 20% glycerol.

Full length human K-Ras4B G12V (GenBank accession number : M54968 for wild type) was expressed using pT7-7 vector as N-terminal hexa-histidin-tagged protein in E. coli JM109. The cultures were suspended in buffer containing 6M guanidin hydrochloride, lOmM Tris-Cl pH8. 0, 1mM MgCl2. The mixture was centrifuged at 40,000 x g for 15min at 4°C. The supernatant was applied onto Ni-NTA- column (QIAGEN) and then 6M guanidin hydrochloride was replaced by 8M urea. K-Ras4B was eluted by pH gradient recommended in the manual of the column, except that buffers contain 1mM MgCl2. The K-Ras containing fractions were pooled and remove urea by gradient dialysis and finally against lOmM Tris-Cl pH8.5 and 1mM MgCl2.

Farnesyl transferase activity is assayed by [3H] farnesylation of recombinant human K-Ras4B protein, by filtration method. 50ul of reaction mixture comprises 50mM Tris-C1 (pH7.5) 5mM dithiothreitol, 0. 3uM ZnCl2, lOmM MgC12, 60nM [1-3H]-farnesyl pyrophosphate (60nCi, Muromachi Yakuhin Kogyo), 86nM K-Ras and 0. 4ug of human farnesyl-protein transferase. Test compounds were dissolved in dimethyl sulfoxide (DMSO) and added to the reaction mixture in a final concentration of less than 1% of DMSO. Reactions were started by addition of [1-3H]-

farnesyl pyrophosphate in 96 well plate. After 60-minute incubation at 30 °C, the reactions are stopped by addition of 50ul of 10% TCA. The mixture is transfer to Multiscreen plate (MAFBNOB50, Millipore), to each well of which 100pal of ethanol are added beforehand. Then the plate was filtered and washed three times with ethanol. 30ul of liquid scintillator OptiPhase SuperMix' (Wallac) was added in each well and the radioactivity was measured on a MicroBeta scintillation counter (Wallac). ICso values were calculated as the concentrations that gave 50% inhibition of farnesyl transferase activity compared with the controls.

Geranylgeranyl-protein transferase activity is assayed by the same method, but the reaction contains 50mM Tris Hcl (pH7.5), 5mM dithiothreitol, 0. 5uM ZnCl2, 10mM MgC12, 5mMATP, 1. 2uM [1-3H]-geranylgeranyl pyrophosphate (3µCi, Muromachi Yakuhin Kogyo), 86nM K-Ras and 7ug of human geranylgeranyl-protein transferase.

Table 2 Inhibition of Farnesyl-protein transferase in the presence of K-Ras Example IC50 (nM) 1 4.5 2 2.4 3 1.9 4 1.6 13 1.0 14 1.2 16 1.1 18 0.9 20 1.0 21 2.5 22 1.1

Example 28 In vitro assay for inhibition of cell growth Antiproliferative activity assay was carried out as follows. A single suspension of tumor cells was inoculated to the serially diluted 96-well microtestplate. Then the template was incubated in the 5% C02 ambience at 37 °C for 4 days (2-3 x 103 cells/well). The degree of cell growth in a monolayer was measured by using WST-8 (Dojindo, Japan). IC50 values of drugs against tumor cells were

calculated as the concentration of drug yielding 50% OD of the control growth. The results are shown in the following table 3.

Table 3 In vitro antitumor activity against human tumor cell line, QG56 Example IC50 (nM) 1 5.9 2 1.1 3 0.6 4 21.0 13 9.6 14 0. 1 16 2.0 18 1.2 20 1.3 21 7.7 22 0.3

Thus the compounds of the invention are therapeutically active, producing regression or remission of solid tumors.

Example 29 The following formulations exemplify typical pharmaceutical compositions in dosage unit form suitable

for systemic or topical administration to warm-blooded animal in accordance with the present invention.

"Active ingredient" (A. I. ) as used throughout these examples refers to a compound of formula (1) and (2), a pharmaceutically acceptable acid or base addition salts or a stereochemically isomeric form thereof.

Example 29-1 Oral solutions 9 g of methyl 4-hydroxybenzoate and 1 g of propyl 4- hydroxybenzoate are dissolved in 4 1 of boiling purified water. In 3 1 of this solution are dissolved first 10 g of 2,3-dihydroxybutanedioic acid and thereafter 20 g of the A. I. The latter solution is combined with the remaining part of the former solution and 12 1 of 1,2, 3-propanetriol and 3 1 of sorbitol 70 % solution are added thereto. 40 g of sodium saccharin are dissolved in 0.5 1 of water and 2 ml of raspberry and 2 ml of gooseberry essence are added.

The latter solution is combined with the former, water is added q. s. to a volume of 20 1 providing an oral solution comprising 5 mg of the A. I. per teaspoonful (5 ml). The resulting solution is filled in suitable containers.

Example 29-2 Capsule formation 20 g of the A. I. , 6 g sodium lauryl sulfate, 56 g starch, 56 g lactose, 0.8 g colloidal silicon dioxide, and 1.2 g magnesium stearate are vigorously stirred together.

The resulting mixture is subsequently filled into 1000 suitable hardened gelatin capsules, each comprising 20 mg of the A. I.

Example 29-3

Film-coated tablet formation A mixture of 100 g of the A. I. , 570 g lactose and 200 g starch is mixed well and thereafter humidified with a solution of 5 g sodium dodecyl sulfate and 10 g polyvinylpyrrolidone in about 200 ml of water. The wet powder mixture is sieved, dried and sieved again. Then there are added 100 g microcrystalline cellulose and 15 g hydrogenated vegetable iol. The whole is mixed well and compressed into tablets, giving 10.000 tablets, each comprising 10 mg of the A. I.

To a solution of 10 g methyl cellulose in 75 ml of denatured ethanol there is added a solution of 5 g of ethyl cellulose in 150 ml of DCM. Then there are added 75 ml of DCM and 2.5 ml 1,2, 3-propanetriol. 10 g of polyethylene glycol is molten and dissolved in 75 ml of DCM. The latter solution is added to the former and then there are added 2.5 g og magnesium octadecanoate, 5 g of polyvinylpyrrolidone and 30 ml of concentrated colour suspension and the whole is homogenated. The tablet cores are coated with the thus obtained mixture in a coating apparatus.

Example 29-4 Injection solution/Emulsion preparation 1.8 g of methyl 4-hydroxybenzoate and 0.2 g of propyl 4-hydroxybenzoate were dissolved in about 0.5 1 of boiling purified water for injection. After cooling to about 50 °C there were added while stirring 4 g lactic acid, 0.05 g propylene glycol and 4 g of the A. I.. The solution was cooled to room temperature and supplemented with water for injection q. s. add 1 1 volume, giving a solution of 4 mg/ml of A. I.. The solution was sterilized by filtration and filled in sterile containers.

Example 29-5 Suppositories 3 g A. I. was dissolved in a solution of 3 g 2,3- dihydroxybutanedioic acid in 25 ml polyethylene glycol 400.

12 g surfactant and 300 g triglycerides were molten. together. The latter mixture was mixed well with the former solution. The thus obtained mixture was poured into moulds at a temperature of 37-38 °C to form 100 suppositories.