CZ448699A3 - Farnesyltransferase inhibitors in combination with HMG CoA reductase inhibitors for treating cancer - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for treatment cancer in a mammal, including a human, comprising an inhibitor Ftase in combination with an HMG CoA reductase inhibitor.

Description

Farnesyltransferase inhibitors in combination with HMG CoA reductase inhibitors for the treatment of cancer

Technical field

The present invention relates to the use of a farnesyltransferase (FTase) inhibitor in combination with a hydroxymethylglutarylcoenzyme A (HMG CoA) reductase inhibitor for the treatment of cancer in mammals.

BACKGROUND OF THE INVENTION

Oncogenes are genes that, when activated, encode protein components of signal transduction pathways that lead to abnormal stimulation of cell growth and mitogenesis. Expression of oncogenes in cultured cells results in cell transformation, which is characterized by the ability of cells to grow in soft agar and the growth of cells as constant foci lacking the contact inhibition exerted by untransformed cells.

Mutation and / or overexpression of certain oncogenes is often associated with human cancers and other disorders involving (e.g., unregulated) cell growth. For example, the growth of benign or malignant tumors may be induced by expression of activated Ras oncogenes or activation of Ras protein by another gene that has undergone oncogenic mutation. Abnormal cell growth that occurs in benign and malignant cells of other proliferative disorders can be induced by aberrant Ras activation. Mutated oncogenic forms of Ras are often found in many human cancers most notably in more than 50% of colon and pancreatic carcinomas (Kohl et al., Science, vol. 260, 1834-1837, 1993).

• · · · · · · · · · · · · · · · · · ·

Ras oncogene is expressed Tip in about 40% of solid malignant tumors that do not respond to conventional chemotherapy.

The K-Ras isoform is expressed in about 90% of pancreatic tumors and about 40% of colorectal and lung cancers.

The Ras-Ras isoform is expressed in about 40% of head and neck cancers.

The Ras-Ras isoform is expressed in most thyroid cancers and in about 20% of acute myeloid leukemias.

To obtain the potential for transformation of normal cells into cancer cells or benign cells that exhibit abnormal growth, as defined below, the Ras oncoprotein precursor must undergo farnesylation of the cysteine residue deposited in the carboxyterminal tetrapeptide. Thus, enzyme inhibitors that catalyze this modification, farnesyl protein transferases, are useful as anticancer agents for tumors in which Ras contributes to transformation.

The K-Ras isoform can be both farnesylated and geranylated in intact cells. Strong farnesyl (FTase) enzyme inhibitors that are highly selective for FTase over geranyltransferase I (GGTase I) may be unable to block the prenylation of the K-Ras mutant and thus ineffective in inhibiting the growth of K-Ras expressing tumor cells.

The present inventor has found that administration of low doses of an HMG CoA reductase inhibitor in combination with a strongly selective FTase inhibitor blocks K-Ras prenylation and K-Ras function as well as Η-Ras prenylation and function. The activity of FTase and GGTase I protein prenyltransferases is dependent on the concentrations of isoprenoid substrates, farnesyl and geranylgeranyl pyrophosphates. Mevalonate is the first intermediate present in the isoprenoid pathway and its synthesis is dependent on HMG CoA reductase activity.

· · Β β β β β β β · · · · · · · β β β β β β β

Compounds such as lovastatin and compactin, which are firmly binding inhibitors of HMG CoA reductase, block the formation of mevalonate and thus block the isoprenoid pathway. Therefore, they inhibit both FTase and GGTase I.

The therapeutic effect of the compounds of these two classes of drugs (FTase inhibitor and HMG CoA reductase inhibitor) is considered to be synergistic. The inventor has found that the combined administration of a FTase inhibitor and a HMG CoA reductase overcomes the limitations that exist when administered separately. It is therefore expected that the combination will be effective in cases where any single agent would not be effective.

Japanese Patent Application JP7316076A, published December 5, 1995, relates to an anticancer pharmaceutical composition containing limonene which, although not a FTase inhibitor, has shown to impair the incorporation of the mevalonic acid-derived isoprene compound into Ras and K-Ras related proteins. , and pravastatin, which is an inhibitor of HMG CoA reductase.

SUMMARY OF THE INVENTION

The present invention relates to a pharmaceutical composition for treating cancer or benign proliferative disorders in a mammal including a human, comprising an FTase inhibitor, an HMG CoA reductase inhibitor and a pharmaceutically acceptable carrier, wherein the active ingredients of the composition (i.e., FTase inhibitor and HMG CoA reductase inhibitor) are present in amounts that make the composition effective in treating cancer or benign proliferative disorders.

The invention also relates to a method of treating cancer or benign proliferative disorders in a mammal, including a human, comprising administering to said mammal an anti-cancer and anti-proliferative effective amount of a pharmaceutical composition comprising a FTase inhibitor, an HMG CoA reductase inhibitor and a pharmaceutically acceptable carrier.

The present invention also relates to a method of treating cancer or a benign proliferative disorder in a mammal including a human comprising administering an FTase inhibitor and an HMG CoA reductase inhibitor in an amount that causes the combination of the two active agents to be effective in treating cancer or benign proliferative disorder.

The invention also relates to a pharmaceutical composition for inhibiting abnormal cell growth in a mammal including a human, comprising an FTase inhibitor, an HMG CoA reductase inhibitor, and a pharmaceutically acceptable carrier, wherein the active ingredients in the composition (i.e., FTase inhibitor and HMG CoA reductase inhibitor) are present in amounts that make the composition effective in inhibiting this abnormal cell growth.

The invention also relates to a method of inhibiting abnormal cell growth in mammals including a human, comprising administering to said human an FTase inhibitor and an HMG CoA reductase inhibitor in amounts that cause the combination of the two active ingredients to be effective in inhibiting this abnormal cell growth.

The term treatment as used herein refers to the prevention, retention or inhibition of the progression of a disorder to which this term is used.

Abnormal cell growth as used herein refers to cell growth that is independent of normal regulatory mechanisms (e.g., loss of contact inhibition).

9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 99 99 99 99 9 9 9 9

- 5 zahrnujeο includes abnormal growth of (1) tumor cells (tumors) expressing activated Ras oncogene, (2) tumor cells in which Ras protein is activated as a result of oncogenic mutation in another gene, and (3) benign and malignant cells of other proliferative diseases in which activation of aberrant Ras occurs.

Examples of such benign proliferative diseases are psoriasis, benign prostatic hypertrophy and restenosis.

Patients who may be treated with a FTase inhibitor in combination with an HMG CoA reductase inhibitor according to the methods of the invention or using pharmaceutical compositions of the invention include, for example, patients who have been diagnosed with lung cancer, bone cancer, pancreatic cancer, skin cancer, head cancer and throat, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, stomach cancer, colon cancer, breast cancer, gynecological tumors (e.g. uterine sarcoma, fallopian cancer, endometrial cancer, cervical cancer, vaginal cancer or genital carcinoma), Hodkin's disease, esophageal cancer, small intestine cancer, endocrine system cancer (e.g., thyroid cancer, parathyroid or adrenal cancer), soft tissue sarcomas, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, t such as childhood tumors, lymphocytic lymphonases, bladder cancer, renal or urethral cancer (e.g., renal cell carcinoma, renal pelvis carcinoma), or central nervous system neoplasms (e.g., primary CNS lymphoma, spinal tumors, brain stem gliomas, or pituitary adenomas).

Patients who can be treated with a FTase inhibitor in combination with an HMG CoA reductase inhibitor according to the method

FFF

F F F F

F F F F

FFF FFF

F F

The FF FF of the invention or the use of the pharmaceutical compositions of the invention may also include patients suffering from abnormal cell growth as defined above.

More specific embodiments of the invention relating to the above pharmaceutical compositions and methods of treatment are those wherein the FTase inhibitor is selected from compounds comprising (a) compounds of Formula I

2 wherein R and R are independently selected from the group consisting of - (CH ₂₎ p (5-10 membered heterocycles), - (CH _2Q aryl), allyl, propargyl .gyl and C-C-alkyl, where p is 0 to 3, wherein said 12 alkyl and alkyl moieties of the R and R groups are optionally substituted with 1 to 3 R substituents, and the aryl and heterocyclic moieties of

R and R groups are optionally substituted with 1 to 3 substituents independently selected from halogen and R 1,

R is - (CH _{2) m} (1- or 2-adamantyl), - _(CH2 ^ (C ^ -CF gCyklo-

X ^4- (A) (B) • «9

9 9 9

999

9

9

9

99

9

99 wherein m is 0-6 and said cycloalkyl and alkyl optionally contain 1 or 2 double or triple bonds,

123

X, X and X are each independently C 1 -C 2 -alkylene optionally containing 1 or 2 double or triple bonds, X is a bond or C 1 -C 2 alkyl optionally containing 1 or 2 double or triple bonds 4 'X bonds and in formula (B), the X moiety is attached to the X moiety at any available carbon in the X' moiety,

R ⁴ is C 8 -C 8 Caryl, 5 to 10 membered heterocyclyl or C 1 -C ⁸ alkyl, each of said R ⁴ groups being optionally substituted with 1 to 3 R ⁸ substituents,

each R substituent is independently selected from the group consisting of halogen, nitro, cyano, phenyl,

C (O) OR ⁶ , -SO ² NR ⁶ R ⁷ , -NR ⁶ R ⁸ , -C (O) R ⁶ , -OR ⁶ , -C (O) NR ⁶ R ⁶ , -OC (O) NR ⁶ R ⁶ , -NR ⁸ C (O) NR ⁸ R ⁶ , -NR ⁸ C (O) R ⁸ , -NR ^and C (O) O (C 1 -C 6 alkyl), -C (NR ⁸ ) NR ⁸ R ⁶ , -C (NCN) NR ⁸ R ⁸ , -C (NCN) S (C 1 -C ₄ alkyl), -NR ^B C (NCN) S (C 1 -C ₄ alkyl), -NR ^e C (NCN) NR ^e R ⁶ , -NR ⁸ SO 2 (C 1 -C ₄ alkyl), -S (O) _n (C 1 -C ₄ alkyl) wherein n is 0 to 2, -NR ⁸ G (O) C (O) NR ⁸ R ⁸ , - NR ⁸ C (O) C (O) R ⁸ , thiazolyl, imidazolyl, oxazolyl, pyrazolyl, triazolyl, tetrazolyl and C 1 -C 6 alkyl, optionally substituted with 1 to 3 fluoro substituents, β 7 each R and R is independently hydrogen or C 1 -C 6 alkyl, each R ⁸ is independently R ⁸ or -OR ⁸ , and each R ⁹ is independently selected from cyano, R ⁸ , -OR ⁸ , -OC (O) R ⁸ , -C (O) OR ⁸ , -C (O) NR ⁸ R ⁷ , -NR ⁸ R ⁷ , -NR ⁸ R ⁸ , -SO ₂ NR ⁸ R ^7, and C 1 -C 6 alkyl substituted with hydroxy, and (b) compounds of formula IIA or IIB; 44 44 ·· »44 4 4 44 · 4 4 4 ··· · 444 ·· ···

4 4 · ·

444 4444 44 44

(HA) (ΙΙΒ) wherein R is hydrogen, halogen (for example chlorine, fluorine, bromine or 555 iodine), cyano, hydroxy, nitro, trifluoromethyl, -NHR, -NR R,

R ⁵ , -OR ^5, or -S (O) -R ⁵ , m

R ² is - (CH _{2) n} -y, or -OCOR ^5,

R is 4-, 3-, or 2-pyridyl, pyrimidyl, pyrazyl,

2-fluoro-4-pyridyl or 3-fluoro-4-pyridyl,

R is 1-adamantyl or 2-adamantyl,

5 5 5 y is hydrogen, hydroxy, amino, cyano, -NHR, -NR R, -NHCOR, -NHCO 3 R ⁵ , halogen, OR ⁵ , -S (O) R ⁵ , -CO 3 H, -CO 3 R ⁵ , -CONR ⁵ R ⁵ , cc Ι Ι 2, j-.

-CONHR, -CONH ₂ , -COR, -CH = CIICO ₂ R, -OCOR, phenyl, phenyl substituted with W, -C = CCO ₂ R ⁵ , -CH = CHR ^5, or -C ^CR ⁵ , each R is independently (C 1 -C 6) branched or branched alkyl, phenyl or benzyl, wherein the phenyl and phenyl portion of benzyl may be optionally substituted by halogen, hydroxy, nitro, cyano, amino, (C 1 -C 6) branched or unbranched alkyl, -C (j) branched or unbranched alkoxy, phenyl, benzyl, (C 1 -C ₄ ) alkylamino, di [(C 1 -C 4) alkyl] amino or -S (O) 2 - (C 1 -C 4) branched or unbranched alkyl,

5 each W is independently halogen, R, hydroxy, -OR 5, nitro, amino, -NHR ⁵ , -NR ⁵ R ⁵ , cyano, or -S (O) _m -R ⁵ ; • · · t · t t 9 t 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 M is 0, 1 or 2, n is 1 to 7, p is 0 or 1,. ,,

E and E are selected independently from the group consisting of hydrogen, halogen, (C 1 -C 6) alkyl, hydroxy, (C 1 -C 6) alkoxy, nitro, trifluoromethyl, cyano, amino, (C 1 -C 6) alkylamino and di [(C 1 -C ₃ ) alkyl] amino, and pharmaceutically acceptable salts thereof,

Het 1 and Het 1 are selected independently from 6-membered heterocyclic rings containing from 1 to 4 nitrogen atoms as part of the ring, optionally substituted with one substituent selected from (C 1 -C 6) alkyl, halogen, hydroxy, (C 1 -C 6) alkoxy, amino, (C 1 ^-C 3) alkylamino and di [(C 1 -C 4) alkyl] amino, and (c) compounds of formula III

(III) where both dashed lines represent possible double bonds,

Z is oxygen or sulfur when double bonded to ring Λ and Z hydroxy, ₍ , kyl-SO ₂ -, adamant-2-yl-S-, naphthyl-S-, benzyl-S-, phenyl-C (= O ) CH ₂ S- (C _{1 -C)} alkyl-OC (= O) _-CH2 -S- or (H, H) (i.e.,

Z represents 2 hydrogen atoms, each of which is simply bonded to the same carbon of ring A) when Z is simply bonded to ring A, and wherein the naphthyl and phenyl and the phenyl moiety of benzyl · may optionally be substituted with from 1 to 3 substituents independently selected from the group consisting of (C 1 -C 6) alkyl optionally substituted with 1 to 3 fluorine atoms, (C 1 -C 6) alkoxy optionally substituted with 1 to 3 fluorine atoms, halogen (e.g. chlorine, fluorine, bromine or iodine), amino, -Cg) alkylamino, di [(C 1 -C 8) alkyl] amino, cyano, nitro, (C 1 -C 8) alkyl-SO _n - wherein n is 0, 1 or 2, -COOH, -COO (C ₁ -C 8) alkyl and -C (O) NH (C 1 -C 8) alkyl,

X is NR ¹ or CHR ¹ ,

R ¹ is hydrogen, (C 1 -C 6) alkyl or (C 1 -C 6) alkylphenyl, wherein ring A is saturated (i.e., when ring A contains no double bond) and R ^x is absent when ring A contains a double bond,

R is selected from the group consisting of naphthyl, phenyl, (C 1 -C 6) alkylphenyl, 1-adamantyl, 2-adamantyl, (C 1 -C 6) branched or unbranched alkyl, (C 1 -C 6) cycloalkyl, and (C 6 -C 6) cycloalkyl. (Cg) bicyclic or tricyclic alkyl, wherein (Cg-Cg) cycloalkyl and (Cg-Cg) bicyclic or tricyclic alkyl may be optionally substituted with hydroxy, and wherein the adamantyl group may be optionally substituted with one to 3 substituents independently selected from (C 1 -C 6) alkyl, halogen and hydroxy; and

R 6 and R 6 are independently selected from benzyl, wherein the phenyl portion of this benzyl may be optionally substituted by amino or nitro, hydrogen, phenyl, (N = C) - (C 1 -C 6) alkyl, (C 1 -C 6) alkyl -OC (= O) - (C 1 -C 6) alkyl and Het-C 1-6 wherein I-Iet is selected from 2-, 3- or 4-pyridinyl, furyl, tetrahydrofuryl, pyrimidyl, pyrazinyl, pyrazolyl, isoxazolyl, thiophenyl and thiazolyl, with the proviso that (a) no more than 1 or 2 dashed lines can represent a double bond in any one compound, (b) when Z is (Η, Η), X is CHgt © ·· © · · © • (C) when Z is oxygen or (11,11) and X is CIIR ''', R ^ must be hydrogen, ( d) when Z is sulfur and X is NR '', R 6 must be hydrogen, and (e) one of R ³ and R ⁴ must be Het-CH ₂ , and

(IV) and pharmaceutically acceptable salts of the foregoing compounds.

Other more specific embodiments of the invention relate to any of the above pharmaceutical compositions and methods of treatment, wherein the FTase inhibitor is selected from compounds 12 of formula I as defined above, wherein R and R are both - (CH 2) z (5-10 membered heterocycles), wherein p is 1 or 2.

Other more specific embodiments of the invention pertain to any of the above pharmaceutical compositions and methods of treatment wherein the FTase inhibitor is selected from compounds of formula I as defined above, wherein R is - (CH ₂ ) (pinane), wherein m is 0 or 2, and in particular those wherein R ³ is pinanmethyl.

Other more specific embodiments of the invention relate to any of the above pharmaceutical compositions and methods of treatment, wherein the FTase inhibitor is selected from compounds 3 of formula I as defined above, wherein R is "·" ···

Where xY X ² , X ² and Χ ^ have the meanings given above; · kde kde kde kde kde kde kde x kde x kde .

Other more specific embodiments of the invention relate to any of the above pharmaceutical compositions and methods of treatment, wherein the FTase inhibitor is selected from compounds of Formula I as described above, wherein iY is phenyl optionally substituted with 1 to 3 R substituents.

Other more specific embodiments of the invention relate to any of the above pharmaceutical compositions and methods of treatment, wherein the FTase inhibitor is selected from the following compounds:

2- [2- (4-bromophenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidin-4-one ,

- [[5-oxo-4,4-bispyridin-4-ylmethyl-1- (2,6,6-trimethylbicyclo [3.1.1- hept-3-ylmethyl) imidazolidin-2-ylidene] acetyl] benzonitrile,

2- [2- (4-chlorophenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidin-4-one ,

2- [2- (3,4-dichlorophenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidine -4-on, 9

9 99 99

99 · 9 99 99 99 9 9 99 9 • 9 9 9 9 9 * 9 999999

9 9 9

9999999 99 99

2- [2- (3-nitrophenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidine -4-on,

2- [2- (4-methoxyphenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidine -4-on,

2- [2- (3-methoxyphenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidine -4-on,

2- [2- (2-methoxyphenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidine -4-on,

2- (2-Biphenyl-4-yl-2-oxoethylidene) -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidin-4-one ,

2- (2-Naphthalen-2-yl-2-oxoethylidene) -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidin-4 -he,

2- [2- (4-fluorophenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidine -4-on,

2- [2- (2,4-Difluorophenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidine -4-on,

4- [[5-oxo-4,4-bispyridin-4-ylmethyl-1- (2,6,6-trimethylbicyclo [3.1.1] hept-3-yl) imidazolidin-2-ylidene] acetyl} benzonitrile , • 9

9

4

4 ·

94 99

9 9 4 ·· «494 • 4,494 49«

9 4

44995 94 49

14- 2- [2- (4-nitrophenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) ) imidazolin-4-one

2- [2-oxo-2-phenylethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidin-4-one,

2- [2-oxo-2- [4- (2H-tetrazol-5-yl) phenyl] ethylidene} -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1. (1) Hept-3-ylmethyl) imidazolidin-4-one;

3- [[5-oxo-4,4-bispyridin-4-ylmethyl-1- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidin-2-ylidene] acetyl] benzonitrile, ethyl ester

4 - [[5-oxo-4,4-bispyridin-4-ylmethyl-1- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidin-2-ylidene] acetyl] benzoic acid ,

2- [2-oxo-2- (4-trifluoromethylphenyl) ethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidine- 4-on,

2- [2- (4-methanesulfonylphenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethyl-3- (2,6,6-trimethylbicyclo [3.1.1] hept-3-ylmethyl) imidazolidin-4 - N - [[1- (6,6-dimethylbicyclo [3.1.1] hept-2-ylmethyl) -5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene] acetyl] benzonitrile,

4 - [(1-bicyclo [2.2.2] oct-1-ylmethyl-5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene) acetyl] benzonitrile,

4- [[1- (2-ethyl-6,6-dimethylbicyclo [3.1.1] hept-3-ylmethyl) -5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene] acetyl] benzonitrile, • ·· «· · · · · · · · · · · · · · · · ·

999 99

99 99 «·

9 · 9 9 9 9 9 9

9 9 9 9 9 · 9,999,999

9 9 9

999 99 99 99

4 - [[1- (2-benzyl-6,6-dimethylbicyclo [3.1.1] hept-3-ylmethyl) -5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene] aceyl} benzonitrile ,

4 - {[1- (2-Isopropenyl-6,6-dimethylbicyclo [3.1.1] hept-3-ylmethyl)]

5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene] acetyl} benzonitrile,

4 - {[1- (2-Isopropyl-6,6-dimethylbicyclo [3.1.1] hept-3-ylmethyl) -5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene] acetyl} benzonitrile - (E 1- [2- (1-methoxyiminoethyl) -6,6-dimethylbicyclo [3.1.1] hept-3-ylmethyl] -5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene } acetyl) benzonitrile,

4 {[1- (6,6-Dimethyl-2-methylenebicyclo [3.1.1] hept-3-ylmethyl) -5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene] acetyl} benzonitrile, - [1- (2-hydroxy-2-hydroxymethyl-6,6-dimethylbicyclo [3.1.1] hept-3-ylmethyl) -5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene] acetyl } benzonitrile,

4- 1- [1- (6,6-dimethyl-2-oxobicyclo [3.1.1] hept-3-ylmethyl) -5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene} acetyl} benzonitrile ,

3-tert-butyl-2- (2-oxo-2-phenylethylidene) -5,5-bispyridin-4-ylmethylimidazolidin-4-one;

4- [1- (2,2-dimethylpropyl) -5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene] acetyl} benzonitrile,

4 - {[1- (2-adamantan-1-ylethyl) -5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene} acetyl} benzonitrile

3-Cyclohexyl-2- (2-oxo-2-phenylethylidene) -5,5-bispyridin-4-ylmethylimidazolidin-4-one;

4 - [(1-adamant-1-ylmethyl-5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene) acetyl] benzonitrile,

4 - [(1-cyclohexylmethyl-5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene) acetyl] benzonitrile,

3-Hexyl-2- (2-oxo-2-phenylethylidene) -5,5-bispyridin-4-ylmethylimidazolidin-4-one,

3-Naphthalen-1-yl-2- (2-oxo-2-phenylethylidene) -5,5-bispyridin-4-ylmethylimidazolidin-4-one;

3-adamantan-1-yl-2- (2-oxo-2-phenylethylidene) -5,5-bispyridin-4-ylmethylimidazolidin-4-one;

3-adamantan-1-yl-2- [2- (4-nitrophenyl) -2-oxoethylidene] -5,5-bispyridin-4-ylmethylimidazolidin-4-one,

4 - [(1-benzyl-5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene) acetyl] benzonitrile,

4 - [(1-allyl-5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylene) acetyl] benzonitrile,

4 - [(1-methyl-5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene) acetyl] benzonitrile,

4- [[1- (2,2-diethoxyethyl) -5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene] acetyl} benzonitrile,

4 - [(1-adamantan-2-ylmethyl-5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene) acetyl] benzonitrile,

4 - [(1-adamantan-2-yl-5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene) acetyl] benzonitrile,

4 [(5-oxo-1-phenyl-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene) acetyl] benzonitrile, and

4- (4-tert-butylphenyl 1-5-oxo-4,4-bispyridin-4-ylmethylimidazolidin-2-ylidene) acetyl] benzonitrile, and pharmaceutically acceptable salts thereof.

Other more specific embodiments of the invention relate to any of the above pharmaceutical compositions and methods of treatment, wherein the HMG CoA reductase inhibitor contained in or used by such a composition is selected from the group consisting of atorvastatin, pravastatin, niacin, gemfibrozil, clofibrate, lovastatin, fluvastatin, simvastatin and compactin; and pharmaceutically acceptable salts of the foregoing compounds.

Other more specific embodiments of the invention relate to any of the above pharmaceutical composition and method of treatment, wherein the HMG CoA reductase inhibitor is contained in or used in such a composition, which is atorvastatin.

Other more specific embodiments of the invention relate to any of the above compositions and methods of treatment, wherein the HMG CoA reductase inhibitor contained in or used in such a composition is lovastatin.

Other more specific embodiments of the invention relate to any of the above pharmaceutical compositions and methods of treatment, wherein the FTase inhibitor contained in or used in such composition is selected from:

(A) compounds of formula (IIA) as defined above, wherein 3;

R is 4-pyridyl, 4-pyrimidyl or 2-fluoro-4-pyridyl, (b) compounds of formula IIA as defined above, wherein R ² is - (CH ₂ ) _n Y, (c) compounds of formula IIA as defined above, wherein R is - (CH 2) Y and n is an integer from 1 to 5, (d) compounds of formula IIA or IIB as defined above, wherein each of R 1, E 2, E ² and R 2, is- when present, it is hydrogen, and (e) compounds of formula IIA as defined above, wherein R is - (CH ₂ ) _n -Y, r 1; is 4-pyridyl, 4-pyrimidyl or 2-fluoro-4-pyridyl, R ⁵ is (C 1 -C 2) alkyl and Y is -CO ² R ⁵ , cyano, -CONHR 4, CII = CHCO ² R ⁵ or -OCOR ⁵ .

Other more specific embodiments of the invention relate to any of the above pharmaceutical compositions and methods of treatment wherein the FTase inhibitor contained in or used in such a composition is not limonene or d-limonene.

The term alkyl as used herein, unless otherwise indicated, includes saturated monovalent hydrocarbon groups having unbranched, branched or cyclic moieties or combinations thereof.

The term halogen as used herein refers to chlorine, fluorine, bromine or iodine.

The above compounds of formulas I, IIA, IIB, III and IV may contain one or more chiral centers and therefore may exist in two or more enantiomeric and diastereoisomeric forms.

9 9 9 99 99 999 99 99

- 19 The above definitions of compounds having the formula I,

,Α, IIB, III and IV include all enantiomers, diastereoisomers and other stereoisomers of these compounds, as well as mixtures thereof.

The following references relate to compounds that exhibit activity as FTase inhibitors and which may be used in combination with an HMG CoA reductase inhibitor, in pharmaceutical compositions and methods of the invention, and to methods of preparation: International Patent Application PCT / US92 / 11292, which determines United States and was public22. U.S. Pat. No. 4,876,259, issued October 24, 1989, U.S. Pat. No. 1345, issued August 2, 1994, U.S. Pat. No. 5,260,332, issued November 9, 1993, U.S. Pat. No. 5,262,435, which was issued

On Nov. 16, 1993, U.S. Pat. No. 5,369,125, issued Nov. 29, 1994, World Patent Application WO 93/24633, published Dec. 9, 1993, World Patent Application WO 94/03597, published Feb. 17, 1994; WO 94/16069, which has been published

June 21, 1994, G. L. Bulton et al., 209. National Meeting of the American Chemical Society, Anaheim, CA, April 2-6, 1995, Division of Med. Chem., Abs. No. 032, World Patent Application

WO 95/00497, published Jan. 5, 1995, US Patent 5,260,479, published November 9, 1993, World Patent Application WO 95/10514, World Patent Application WO 95/10515, World Patent Application WO 95/10516 World Patent Application WO 95/12572, which has been published

May 11, 1995, World Patent Application WO 95/11917, published May 4, 1995, World Patent Application WO 94/26723, published Nov. 24, 1994, World Patent Application WO 95/25086, published Nov. 21, 1995. September 1995.

«· ·· · · · · · · · · · · · · · . · *. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. .. ..

- 20 Next is Kanda et al., AFMC, International Symposium of Medical Chemistry AIMECS 95, Tokyo, Japan, Poster P7M153,

September 4, 1995, World Patent Application WO 96/10037, published April 4, 1996, World Patent Application WO 96/10035, published April 4, 1996, World Patent Application WO 96/10034, published

On April 4, 1996, World Patent Application WO 96/10011, published April 6, 1996, World Patent Application WO 96/10011, published April 6, 1996, World Patent Application WO 96/09821, published

On April 4, 1996, World Patent Application WO 96/09820, published April 4, 1996, Quin et al., 211. National Meeting of the American Chemical Society, New Orleans, LA, 24.28. March 1996, lecture COMP 012, March 24, 1996, World Patent Applications WO 96/06609 and WO 96/06604, both published on March 7, 1996, European Patent Application EP 696 593, published February 14, 1996, Hartman , GD, 14th International Symposium of Medical Chemistry, Maastricht, Netherlands, 8.-12. September 1996, lecture SL-08.3, 10.9.96, World Patent Application WO 96/30363, published October 3, 1996, World Patent Application WO 96/30343, published October 3, 1996, World Patent Application WO 97 / No. 03050, World Patent Application WO 94/26723, published November 24, 1994, International Patent Application PCT / IB 95/00189, which designates the United States, and filed March 20, 1995, US Patent Application 08/236 743, which was filed on April 29, 1994, U.S. provisional application entitled Adamantyl substituted oxindoles as pharmaceutical compositions, filed May 28, 1996 in the name of RAVolkmann and JPLyssikatos, U.S. Patent 5,350,867, issued September 27, 1994, U.S. Patent 5,352 No. 705, issued October 4, 1994; U.S. Patent 5,565,489, issued October 15, 1996; European Patent Application EP 750,609, published and 2 January 1997, European Patent Application 461,869, published Dec. 18, 1991, " 9

I 9 9 99 8

- 21 and World Patent Application 96/21456, published July 18, 1996.

The following references apply to compounds that exhibit activity as HMG CoA reductase inhibitors and which can be used in combination with a FTase inhibitor in the pharmaceutical compositions and methods of the invention, and to methods for their preparation: US Patent 4,681,893, issued July 21, 1987, U.S. Patent 5,273,995, issued December 28, 1993,

U.S. Patent 5,385,929 issued January 31, 1995, U.S. Patent 4,957,971 issued September 18, 1990, U.S. Patent 5,102,893 issued April 7, 1992, U.S. Patent 4,957,940 issued September 18, 1990, U.S. Pat.

No. 950,675, issued August 21, 1990; U.S. Patent 4,929,620 issued

May 29, 1990, U.S. Patent 4,923,861, issued May 8, 1990,

U.S. Patent 4,906,675 issued March 6, 1990, U.S. Patent 4,868,185 issued September 19, 1989, U.S. Patent 5,124,482 issued June 23, 1992, U.S. Patent 5,003,080 issued March 26, 1991, U.S. Pat.

No. 097,045, issued March 17, 1992, U.S. Pat. No. 5,149,837 issued

September 22, 1992, U.S. Patent 4,906,624, issued March 6, 1990,

U.S. Patent 4,761,419, issued August 2, 1988, U.S. Patent 4,735,950, issued April 5, 1938, U.S. Patent 4,808,621, issued February 28, 1989, U.S. Patent 4,647,576, issued March 3, 1987, U.S. Pat. June 2, 1992, U.S. Patent 5,214,197 issued

May 25, 1993, U.S. Patent 5,321,046, issued June 14, 1994,

U.S. Patent 5,260,440 issued November 9, 1993 and U.S. Patent 5,208,258 issued May 4, 1993.

Further, it is U.S. Pat. No. 5,369,125 issued November 29, 1994, U.S. Pat. No. H1345 issued August 2, 1994, U.S. Pat. No. 5,262,435, issued November 16, 1993, and U.S. Pat.

Further, it is British Patent Application GB 2,055,100 published February 25, 1981, U.S. Patent 4,499,289 issued

February 12, 1983, U.S. Patent 4,645,854, issued February 24, 1987,

U.S. Patent 4,613,610 issued September 23, 1986; U.S. Patent 4,668,699

Φ • φ r

♦ Φ · · ·

26. · Φ Φ issued May 26, 1987, US patent 4,851,436 issued July 25, 1989 US patent 4,678,806 issued July 7, 1987, US patent 4,772,626 issued September 20, 1988, US patent 4,855,321 issued

August 8, 1989, European Patent Application EP 244 364 published November 4, 1987, US Patent 4,766,145 issued August 23, 1988, US Patent 4,876,279 issued October 24, 1989, US Patent 4,847,306 issued July 11, 1989, U.S. Patent 5,049,696 issued September 17, 1991, European Patent Application EP 245,990 published November 19, 1987, European Patent Application EP 251,625 published January 7, 1988, US Patent 4,719,229 Published January 12, 1988, and Japanese Patent Application 63014722 published January 21, 1988.

Further, U.S. Pat. No. 4,736,064, issued April 5, 1988,

U.S. Patent 4,738,982, issued April 19, 1988;

845,237 issued July 4, 1989, European Patent EP 306,263 issued March 18, 1992, US Patent 5,026,708 issued June 25, 1991, US Patent 4,863,957 issued September 5, 1989, US Patent 4,946,841 issued August 7, 1990 , European Patent 339,358 issued July 13, 1994, U.S. Patent 4,937,264, issued June 26, 1998, U.S. Patent 4,876,366, issued October 24, 1989, U.S. Patent 4,921,974, issued May 1, 1990, U.S. Patent 4,963,538, issued On October 16, 1990, U.S. Patent 5,130,306, issued July 14, 1992, U.S. Patent 4,900,754, issued February 13, 1990, U.S. Patent 5,026,698, issued June 25, 1991, U.S. Patent 4,977,161, issued December 11, 1990, U.S. Pat. U.S. Patent 4,927,851 issued May 22, 1990, European Patent Application EP 373,507 published June 20, 1990, US Patent 4,939,143 issued July 3, 1990, US Patent 4,939,159 issued July 3, 1990, US Patent 4,940,727 July 10, 1990, U.S. Patent 5,116,870, issued May 26, 1992; Australian Patent AU 635,545, issued March 25, 1919; 93, U.S. Pat. No. 5,098,391, issued Mar. 24, 1992, U.S. Pat

No. 294,724, issued March 15, 1994; U.S. Patent 5,001,255, issued March 19, 1991; U.S. Patent 5,149,834, issued September 22, 1992;

U.S. Patent 5,089,523, issued February 18, 1992, European Patent Application EP 465,265, issued January 8, 1992, U.S. Patent 5,476,846, issued December 19, 1995, U.S. Patent 5,321,046, issued June 14, 1994, U.S. Patent 5 No. 106,992, issued April 21, 1992; and U.S. Patent 5,347,039, issued September 13, 1994.

Further, Japanese Patent Application 4193836, published July 13, 1992, British Patent Application 2253787 published September 23, 1992, US Patent 5,411,969, issued May 2, 1995, Japanese Patent Application 4,356,435, published December 10, 1992, US Patent 5 266 707 issued

November 30, 1993, U.S. Patent 5,455,247 issued October 3, 1995, U.S. Patent 5,475,029 issued December 12, 1995, U.S. Patent 5,591,772 issued January 7, 1997, U.S. Patent 5,286,746 issued February 15, 1994, Japanese Patent Application JP 7089898 published April 4, 1995, European Patent Application EP 677 039 published October 18, 1995 and World Patent Application 96/08248 published March 21, 1996.

The invention relates both to methods of treating cancer wherein the FTase inhibitor and the HMG CoA reductase inhibitor are co-administered as part of the same pharmaceutical composition, as well as methods wherein the two active agents are administered separately as part of an appropriate dosing regimen designed to obtain the benefits of combination therapy. . The appropriate dosing regimen, the amount of each dose administered and the specific intervals between doses of each active agent will depend on the subject being treated, the type of cancer or abnormal cell growth, and the severity of the condition.

In practicing the methods of the present invention, the FTase inhibitor will be administered in amounts reported in the literature or in amounts believed to be effective for administering the compound as the only active agent for the present invention.

9 9 9 9

9 * · · · · ······ · · 9 · 9 «··· · · 9 9 9 9- · 9 9 © · ··

Treating cancer or inhibiting abnormal cell growth, and the HMG CoA reductase inhibitor will be administered in an amount that is about 1/4 to 1/2 of the amount reported in the literature or otherwise believed to be effective for administering such a compound as the only agent for the treatment of hypercholesterolemia.

For example, in the practice of the present invention, FTase inhibitors of Formulas I, IIA, IIB and III will typically be administered to an average 70 kg heavy adult human in an amount ranging from about 0.005 to about 0.6 mg / kg body weight of a subject being treated per day. atorvastatin HMG CoA reductase inhibitor will typically be administered in an amount ranging from about 0.07 to about 3.6 mg per kg of body weight per day, in single or divided doses.

However, there may be differences depending on the species of animal to be treated and their individual responses to said drug as well as the type of pharmaceutical formulation chosen and the time period and interval at which such administration is performed. In some cases, dosing levels below the lower limit of the above range may be more than adequate, while in other cases dosing levels higher than the above daily limit may be used without causing any harmful side effect provided such higher doses are administered as several small doses for administration during the day.

The FTase inhibitors and HMG CoA reductase inhibitors used in the pharmaceutical compositions and methods of the invention are also hereinafter referred to as therapeutic agents. The therapeutic agents may be administered either by the oral or parenteral routes. Compositions comprising both a FTase inhibitor and an HMG CoA reductase inhibitor will generally be administered orally or parenterally daily in single or divided doses so that the total amount of each active agent administered falls within the above guidelines.

· FFF FFF FFF FFF FFF FFF FFF FFF FFF FFF

The therapeutic compositions may be administered alone or in combination with pharmaceutically acceptable carriers or diluents by any of the routes previously indicated, and such administration may be in single or multiple doses. In particular, the novel therapeutic compositions of the invention may be administered in a wide variety of different dosage forms, that is, combined with various pharmaceutically acceptable inert carriers in the form of tablets, capsules, pills, lozenges, hard lozenges, suppositories, aqueous suspensions, injectable solutions, elixirs, syrups and the like. Such carriers include solid diluents or fillers, sterile aqueous media and various non-toxic organic solvents, and so on. In addition, oral pharmaceutical compositions may be suitably sweetened and / or flavored. Generally, the therapeutic compounds of the invention, when administered separately, i.e. not in the same pharmaceutical composition, are present in such dosage forms at concentration levels ranging from about 5.0% to about 70% by weight.

For oral administration, tablets containing various excipients such as microcrystalline cellulose, sodium citrate, calcium carbonate, dicalcium phosphate and glycine may be used together with various disintegrants such as starch (and preferably corn, potato or tapioca starch), alginic acid and various complex silicates , together with granulating binders such as polyvinylpyrrolidone, sucrose, gelatin and acacia resin. In addition, lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc are often very useful for tableting purposes.

Solid compositions of a similar type may also be employed as fillers in gelatin capsules, with preferred agents

9 999

98 • 9 9 9

9 9 9

999 999

9

Also in this connection are lactose or milk sugar as well as high molecular weight polyethylene glycols. When aqueous suspensions and / or elixirs for oral administration are desired, the active ingredient may be combined with various sweetening or flavoring, coloring or coloring agents and, if desired, emulsifying and / or suspending agents, as well as together with such diluents as water, ethanol, propylene glycol, glycerin and various combinations thereof.

For parenteral administration, solutions of the therapeutic agent in either sesame or peanut oil or in aqueous propylene glycol may be used. The aqueous solutions should be suitably buffered if necessary and the liquid diluent first act isotonic. These aqueous solutions are suitable for intravenous injection purposes. The oily solutions are suitable for intraarticular, intramuscular and subcutaneous injection purposes. The preparation of your solutions under sterile conditions is readily accomplished by standard pharmaceutical technologies well known to those skilled in the art.

The activity of therapeutic compounds as FTase inhibitors can be determined by their ability to control FTase inhibition in vitro. This procedure is described below.

The crude FTase preparation containing the cytosolic fraction of homogenized brain tissue was used to screen compounds in a 96-well assay plate. Cytosol fractions are prepared by homogenizing approximately 40 g of fresh tissue in 100 ml of sucrose / t-IgC4 / EDTA buffer (using a Dounce homogenizer, up to 15 strokes), centrifuging the homogenates at 1000 g for 10 minutes at 4 G, re-centrifuging the supernatant at 17 000 g for 15 minutes at 4G and then collecting the resulting supernatant.

• * * »·» »

9 9 9 9 9 9 9 9 · 999 9

9 · · · · * 9 9 99 9 9 999 99 99

This supernatant is diluted to contain a final concentration of 50 mM Tris-HCl (pH 7.5), 5 mM DTT, 0.2 M HCl, 20 mM ZnCl ₂ , mM PMCF and re-centrifuged at 178,000 g for 90 minutes. at 4G. The supernatant, labeled crude FTase, was assayed for protein concentration, aliquoted and stored at -70 ° C.

The assay used to measure in vitro inhibition of human FTase is a modification of the method described by Amersham LifeScience for the use of their farnesyltransferase (3H) Scintilation Proximity Assay (SPA) kit (TRKQ 7010).

FTase enzyme activity is determined in a volume of 100 ml containing 50 mM N- (2-hydroxyethyl) piperazine-N- (2-ethanesulfonic acid) (HEPES, pH 7.5, 30 mM MgCl ₂ , 20 µM KCl, mM Na 2 -). IPO 2, 5 mM dithiothreitol (DTT), 0.01% Triton X100, 5% dimethylsulfoxide (DMSO), 20 mg crude FTase, 0.12 mM [3 H] -farnesylpyrophosphate ([3 H] -FPP, 36,000 dpm / pmol, Amersham LifeScience) and 0.2 mM biotinylated Ras peptide KTKCVIS (Bt-KTKCVIS), which is M-terminally biotinylated in its alpha-amino group and was synthesized and purified by HPLC in the laboratory. reagent in the TRKQ 7010 kit), followed by a 45 minute incubation at 37 ° C.

The prenylated and unphenylated Bt-KTKCVIS were captured by adding 10 ml of steptavidin-coated SPA beads (TRKQ 7010) per well and incubating the reaction mixture for 30 minutes at room temperature. The amount of radioactivity bound to the SPA beads is determined using a MicroBeta 1450 board computer.

Under these assay conditions, enzyme activity is linear with respect to prenyl group acceptor, Bt-KTKCVIS and crude FTase concentrations, but saturating with respect to prenyl donor, FPP. The reaction time of the assay is also within the linear range.

99 9 99 9 9 9 9 9 9 9 9 999 999

9 · 9 · 9 9

999 99 999 9999

Test compounds are routinely dissolved in 100% DMSO. Inhibition of farnesyltransferase activity is determined by calculating the percentage incorporation of tritiated farnesyl in the presence of test compound versus its incorporation in control wells (absence of inhibitor). IC 50 values, i.e., the concentration required to produce half of the maximum farnesylation of Bt-KTKCVIS, are determined from the dosing responses obtained.

A fluorescence assay for FTase activity that can be used to screen an FTase inhibitor is described in UK Patent Application GB 2,267,966, published December 22, 1993.

The activity of certain therapeutic agents as HMG CoA reductase inhibitors can be determined as described by Dugan et al., Achiv. Biochem. Biophys., 1972, 152, 21-27.

In this method, the level of HMG-CoA enzyme activity in standard rats is determined by feeding the rats with a diet containing 5% cholestylamine for 4 days, after which the rats are sacrificed. The liver of the rats is homogenized and the incorporation of cholesterol-C-acetate into the unsaponifiable lipid in the rat liver homogenate is measured. The micromolar concentration of the compound required for 50% inhibition of sterol synthesis over a one-hour period is measured and expressed as an IC 50 -q value.

The second method (referred to as COR sorting) is that described by T-Kit et al., J. Clin. Invest., (1980, 66: 10941100). In this method, the amount of? C-HMG-CoA converted to? C-mevalonate is measured in the presence of purified HMG-CoA reductase enzyme addition. The micromolar concentration of the compound required for 50% inhibition of cholesterol synthesis is measured and recorded as an IC 50 value.

·· ··

9 9 ·

9 9 ♦

99,999

9

99

The various methods of the invention can be practiced as part of a therapy that involves administering one or more other antitumor agents, for example those selected from mitotic inhibitors such as vinblastine, alkylating agents such as cisplatin, carboplatin and cyclophosphamide, antimetabolites such as 5-fluorouracil , cystosinarabinosy and hydroxyureas or, for example, one of the preferred antimetabolites disclosed in European Patent Application No. 239,362 such as N- {5- [n- (3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino] -2-thenoyl-4-L-glutamic acid, intercalar antibiotics such as adriamycin and bleomycin, enzymes such as asparaginase, topoisomerase inhibitors such as etoposide, biological response modifiers such as interferon and antihormones such as antiestrogens such as NOLVADEX (tamoxifen) or antiandrogens such as CASODEX (4'-cyano-4- (4-fluorophenylsulfonyl) 12-Hydroxy-2-methyl-3 '- (trifluoromethyl) propionanilide.

Such therapies may be achieved by way of simultaneous, sequential or separate dosing of the individual components of the therapy. According to this aspect of the invention, there is provided a pharmaceutical product comprising a pharmaceutically acceptable carrier as described above, one or both HMG CoA reductase and FTase inhibitors, and another anti-tumor agent as described above.

As indicated in Table 1 below, the invention has shown that the activity of Compound 1 having a structure

«» · ·· ·· 99 9 9 9 9 9

9

9 9 999999

9 9 9

999 99 99 99

- 30 may be increased by a minimum effective dose of lovastatin.

Table 1

Synergistic effects of lovastatin and therapeutic compound I on K-Ras 4B prenylation in intact cells

% inhibition of OF K-Ras 4B prenylation ^{X //} Compound 1 (µM) control + 5 µM lovastatin 0 0 23 0.1 0 56 1/0 0 83 10 0 96

x / semi-confluent monolayers of NIH-3T3 transfectant overexpressing mutant K-Ras 4B were treated for 18 hours at 38 ° C with increasing concentrations of CP-390 392 in the presence and absence of 5 µl of hydrolyzed lovastatin. Cells were lysed in RIPA lysis buffer (50 µM tris [hydroxymethyl iaminomethane, 0.15 M sodium chloride, 1% sodium deoxychot, 1% Triton X-100, 0.1% SDS, 0.25 sodium azide, pH 8.5). containing 1 mM DTT (dithiothreitol, Boehringer Mannheim, Indianapolis, IN) and protease inhibitors (aprotinin, leupeptin, anidpain, pefabloc at final concentrations of 10 µg / ml, 2 µg / ml, 2 µg / ml and 50 µm respectively, Boehringer Mannheim , Indianopolis, IN) and cooked for 3 minutes.

• ···· © © · · · ··· ··· • ©

Equal amounts of protein (100 µm / lane) were digested by SDS-PAGE on 12.5% gels and transferred to ImmobilonP membranes (Intergrated Separation Systems, Natick, MA).

These membranes were immunoblotted with 5 microg / ml anti-Pan-ras (Ab-3) monoclonal antibody (Calbiochem, La Jolla, CA). The spots were incubated with peroxidase-conjugated secondary antibody and immunoblotted with Ras protein determined by increased chemiluminescence (Amersham Life Products, Arlington, Heights, IL). The percentage of phenylated Ras was determined by densitometric scanning using MasterScan 3.0 (Scanalytics, Billerica, Massachusettes). -

Claims (9)

A pharmaceutical composition for treating cancer or benign proliferative disorders in a mammal comprising an FTase inhibitor, an HMG CoA reductase inhibitor, and a pharmaceutically acceptable carrier, wherein the FTase inhibitor and the HMG CoA reductase inhibitor are present in amounts that make the composition effective in treating cancer or benign proliferative disorders. A pharmaceutical composition according to claim 1 wherein the FTase inhibitor is selected from compounds comprising (a) compounds of formula I wherein R and R are independently selected from the group consisting of - (CII ₂ ) (5-10 membered heterocycles), - (CII ₂ ) p (Cg-C ^ ₀ aryl), allyl, proparrgyl and C-C, _a alkyl, wherein p is 0 to 3, wherein said ^{X 0} 12 alkyl and alkyl portions of the R and R groups are optionally substituted with 1 to 3 R substituents, and aryl and heterocyclic portions 1 2 R and R groups are optionally substituted with 1 to 3 substituents independently selected from halogen and R 1, R ⁵ is - (CH ₂ ) _m (1- or 2-adamantyl), - (CH ₂ ) - (CH ₂ ) -cyclo- (A) (B) II • 99 * • 99 9 «9 9 9 9 © 9 9 9 9 9 99 99 9 9 9 9 9 9 9 9 9 · 9,999,999 • 9,9 9 9 99 99 99 wherein m is 0-6 and said cycloalkyl and alkyl optionally contain 1 or 2 double or triple bonds, 12 3 v X, X and X are each independently C 1 -C ₇ alkylene, optionally containing 1 or 2 double or triple bonds, X is a bond or C 1 -C 2 -alkyl optionally containing 1 or 2 double or triple 1/4 1 in> the bond and in formula (B), the X moiety is attached to the X moiety in any available carbon in the X '' moiety, R ⁴ is C _r -C _in -aryl, 5-membered heterocyclyl or to C.-C _y alkyl, wherein each of said R group is optionally sub1 HV 5 The substituted 1 to 3 R substituents 5 of each R substituent is independently selected from the group consisting of halogen, nitro, cyano, phenyl, C (O) OR ^8, -SO 2 NR ⁸ R ^7, -NR ⁸ R ^8, -C (O) R ^e, -OR ^6, -C (O) NR ^S R ^8, -OC (O) NR ⁸ R ⁸ , -NR ⁸ C (O) NR ⁸ R ⁸ , -NR ⁸ C (O) R ⁸ , -NR ⁸ C (O) O (C 1 -C 4 alkyl), -C (NR ⁸ ) NR ⁸ R ⁸ , -C (NCN) NR ^B R ^e , -C (NCN) S (C 1 -C 4 alkyl), -NR ⁸ C (NCN) S (C 1 -C ₄ alkyl), -NR ⁸ C (NCN) NR ^B R ⁶ , -NR ⁸ SO ₂ (C 1 -C 4 alkyl), -S (O) _n (C 1 -C ₄ alkyl) wherein n is 0 to 2, -NR ⁸ C (O) C (O) NR ⁸ R ⁸ -NR ⁸ C (O) C (O) R ⁸ , thiazolyl, imidazolyl, oxazolyl, pyrazolyl, triazolyl, tetrazolyl and C 1 -C 6 alkyl, optionally substituted with 1 to 3 fluoro substituents, Γ * 7 ⁱⁿ each R and R 'is independently hydrogen or C ^ -C ^ alkyl, each R ⁸ is independently R ⁸ or -OR ⁸ and each R 9 is independently selected from cyano, R ^8, -OR ^8, -OC (O) R ⁸ , -C (O) OR ⁸ , -C (O) NR ⁶ R ⁷ , -NR ⁸ R ⁷ , -NR ⁸ R ⁸ , -SO ₂ NR ⁶ R ^7, and C 1 -C ⁶ alkyl and (b) compounds of formula IIA or IIB - III F FF FF FFF FF FFF FF FFF FF FFF FF FFF FF FFF FF FF · FFFF ·· FF or (IIA) (IIB) wherein R is hydrogen, halogen (for example chlorine, fluorine, bromine or 5 5 5 iodine), cyano, hydroxy, nitro, trifluoromethyl, -NHR, -NR R, R ⁵ , -OR ^5, or -S (O) -R ⁵ , m R ² is - (objective _{2) n} -y, or -OCOR ^5, R is 4-, 3-, or 2-pyridyl, pynmidyl, pyrazmyl, 2-fluoro-4-pyridyl or 3-fluoro-4-pyridyl, R is 1-adamantyl or 2-adamantyl, 5 5 5 5 y is hydrogen, hydroxy, amino, cyano, -NHR, -NR R, -NHCOR, -NI-R ⁵ , halogen, OR ⁵ , -S (O) R ⁵ , -CO ₉ II, -CO R ⁵ , -CONR ⁵ R ⁵ , Z j- C tli p · Z J- Λ * -CONIIR, -CONH ₂ , -COR, -CH = CIICO ₂ R, -OCOR, phenyl, phenyl substituted with W, -C = CCO ₂ R ⁵ , -CH = CI ₁ R ⁵ or -C = CR ⁵ , each R is independently (C 1 -C 6) unbranched or branched alkyl, phenyl or benzyl, wherein the phenyl and phenyl portion of the benzyl may be optionally substituted by halogen, hydroxy, nitro, cyano, amino, (C 1 -C 6) branched or unbranched alkyl, -C ₄ ) branched or unbranched alkoxy, phenyl, benzyl, (C 1 -C ₄ ) alkylamino, di [(C ₁ -C ₄ ) alkyl] amino or -S (O) _m - (C ₁ -C ₄ ) branched or unbranched alkyl , Each W is independently halogen, R, hydroxy, -OR 5, nitro, amino, -NHR ⁵ , -NR ⁵ R ⁵ , cyano or -S (O) _m -R ⁵ ; 99,999 - IV in is 0, 1 or 2, n is 1 to 7, p is 0 or 1, E and E are selected independently from the group consisting of hydrogen, halogen, (C 1 -C 6) alkyl, hydroxy, (C 1 -C 6) alkoxy, nitro, trifluoromethyl, cyano, amino, (C 1 -C 6) alkylamino and diJAC (C 1 -C 4) alkyl] amino, and pharmaceutically acceptable salts thereof, Het 1 and Iet 1 are independently selected from 6-membered heterocyclic rings containing from 1 to 4 nitrogen atoms as part of the ring, optionally substituted with one substituent selected from (C 1 -C ₃ ) alkyl, halogen, hydroxy, (C 1 -C 4) alkoxy, amino , (C 1 -C ₃ ) alkylamino and di) alkyl] amino, and (c) compounds of formula III (III) wherein both dashed lines represent optional double bonds, oxygen or sulfur, when hydroxy, is double bonded to ring A and ^{(C c io) alkyl S0} '^"(Cl" ^{C 10) alkyl-SC>} ₂ -, adamant-2-yl-S-, naphthyl-S-, benzyl-S-, phenyl-C (= O) CH ₂ -S-, (C ₁ -C ₆₎ alkyl-OC (= O) _-CH2 -S- or (H, H) (i.e., Z represents 2 hydrogen atoms, each of which is simply bound to the same carbon of ring A) when Z is simply bound to ring A, and wherein the naphthyl and phenyl and the phenyl moieties of benzyl are May be optionally substituted with from 1 to 3 substituents independently selected from the group consisting of (C 1 -C 6) alkyl optionally substituted with 1 to 3 fluorine atoms, (C 1 -C 6) alkoxy optionally substituted with 1 to 3 substituents; up to 3 fluorine atoms, halogen (e.g. chlorine, fluorine, bromine or iodine), amino, (C 1 -C 6) alkylamino, di [(C 1 -C 6) alkyl] amino, cyano, nitro, (C 1 -C 6) alkyl-SO _n - wherein n is 0, 1 or 2, -COOH, -COO (C 1 -C 6) alkyl and -CONONH 2 (C 1 -C 6) alkyl, X is NR ¹ or CHR ¹ , R ¹ is hydrogen, (C ¹ -C 6) alkyl or (C 1 -C 6) alkylphenyl, wherein ring A is saturated (i.e., when ring A contains no double bond) and R ¹ is absent when ring A contains a double bond, R is selected from the group consisting of naphthyl, phenyl, (C1-C6) alkylphenyl, 1-adamantyl, 2-adamantyl, (C1-C6) branched or unbranched alkyl, (C1-C6) cycloalkyl and (C1-C6) (g) a bicyclic or tricyclic alkyl, wherein the (C 1 -C 6) cycloalkyl and the (C 6 -C 6) bicyclic or tricyclic alkyl may be optionally substituted with a hydroxy group, and wherein the adamantyl group may be optionally substituted with one to 3 substituents independently selected from ( C1-C6 alkyl, halogen and hydroxy, and R ¹ and R ⁴ are independently selected from benzyl, wherein the phenyl portion of this benzyl may be optionally substituted with amino or nitro, hydrogen, phenyl, (N = C) - (C 1 -C 6) alkyl, (C 1 -C 6) alkyl-O- C ^ Oj-ÍCpCg) alkyl and Het-CH _2, wherein Ilet is selected from 2-, 3- or 4-pyridinyl, furyl, tetrahydrofuryl, pyrimidyl, pyrazinyl, pyrazolyl, isoxazolyl, thiophenyl and triazolyl, provided that (a ) no more than 1 or 2 dashed lines may represent a double bond in any one compound, (b) when Z is (II, Η), X is CH ₂ , · ··· · ··· • · · ···· ·· · · VI (c) when Z is oxygen or (II, II) and X is ClIR ^, R ^ must be hydrogen, (d) when Z is sulfur and X is NR ^, R ^ must be hydrogen, and (e) one za must be Het-CH ₂ , and CH, (IV) and pharmaceutically acceptable salts of the foregoing compounds AND. The pharmaceutical composition of claim 1 wherein the HMG CoA reductase inhibitor is selected from the group consisting of atorvastatin, pravastatin, lovastatin, compactin, fluvastatin and simvastatin, and pharmaceutically acceptable salts of the foregoing compounds. A method of treating cancer or a benign proliferative disorder in a mammal, comprising administering to the mammal a pharmaceutical composition according to any one of claims 1 to 3. 5. A method of treating cancer or a benign proliferative disorder in a mammal comprising administering to the mammal an FTase inhibitor and an HMG CoA reductase inhibitor, wherein the FTase inhibitor and the HMG CoA reductase inhibitor are administered in amounts that make the combination of the two active agents effective in treating cancer. or benign proliferative disorders. - VII The method of claim 5, wherein the HMG CoA reductase inhibitor is atorvastatin, pravastatin, fluvastatin, simvastatin, lovastatin or compactin, or a pharmaceutically acceptable salt thereof. 7. A pharmaceutical composition for inhibiting abnormal cell growth in mammals comprising an FTase inhibitor, an HMG CoA reductase inhibitor, and a pharmaceutically acceptable carrier, wherein the active ingredients in the composition are present in amounts that render the composition effective to inhibit abnormal cell growth. 8. A method of inhibiting abnormal cell growth in a mammal comprising administering to a mammal with abnormal cell growth inhibiting an effective amount of a pharmaceutical composition comprising an FTase inhibitor, an HMG CoA reductase inhibitor, and a pharmaceutically acceptable carrier. 9. A method of inhibiting abnormal cell growth in a mammal comprising administering to the mammal an FTase inhibitor and an inhibitor HMG CoA reductase in amounts that make the combination of the two active agents effective in inhibiting abnormal cell growth.