AU735366B2 - Inhibitors of farnesyl protein transferase - Google Patents

Description

WO 99/18951 PCT/US98/19315 INHIBITORS OF FARNESYL PROTEIN TRANSFERASE Field of the Invention This invention relates to compounds thatinhibit farnesylprotein transferase and ras protein farnesylation, thereby making them useful as anti-cancer agents. The compounds are also useful in the treatment of diseases, other than cancer, associated with signal transduction pathways operating through ras and those associated with proteins other than ras that are also post-translationally modified by the enzyme farnesyl protein transferase. The compounds may also act as inhibitors of other prenyl transferases, and thus be effective in the treatment of diseases associated with other prenyl modifications of proteins.

Background of the Invention The mammalian ras gene family comprises three genes, H-ras, K-ras and N-ras. The ras proteins are a family of GTP-binding and hydrolyzing proteins that regulate cell growth and differentiation.

Overproduction of normal ras proteins or mutations that inhibit their GTPase activity can lead to uncontrolled cell division.

The transforming activity of ras is dependent on localization of the protein to plasma membranes. This membrane binding occurs via a series of post-translational modifications of the cytosolic Ras proteins.

The first and mandatory step in this sequence of events is the farnesylation of these proteins. The reaction is catalyzed by the enzyme farnesyl protein transferase (FPT), and farnesyl pyrophosphate (FPP) serves as the farnesyl group donor in this reaction. The ras C-terminus contains a sequence motif termed a "Cys-Aaal-Aaa2-Xaa" box (CAAX box), wherein Cys is cysteine, Aaa is an aliphatic amino acid, and Xaa is a serine or methionine. Farnesylation occurs on the cysteinyl residue of the CAAX box (cys-186), thereby attaching the prenyl group on the protein via a thio-ether linkage.

WO 99/18951 PCT/US98/19315 2 Brief Description of the Invention In accordance with the present invention, compounds of the formulas I, and II and R6 RG-RrSrTt Y-

R

1

,R

2

,R

3 their enantiomers, diastereomers, and pharmaceutically acceptable salts, prodrugs and solvates thereof inhibit farnesyl protein transferase which is an enzyme involved in ras oncogene expression. In formulas I- II and throughout their specification, the above symbols are defined as follows: m, r, s and t are 0 or 1; 7 X is selected from the group consisting of carbon, oxygen, NR or absent.

Z is selected from the group consisting of CHR 9

SO

2 CO, C02, O, NR 10 2 0 SO2NR 1 1

CONR

1 2 WO 99/18951 PCT/US98/19315 3 N 'CN N -CN0 11 11 11 v-C-N- r 1 -N-SO 2 11 1 1411 0 11 20 22 Iv -S0 2 0 1 NR R 'N NR 2

R

16

R

1

R

18

R

19 S- I or Z may be absent; Y is selected from the group consisting of CHR 23 so 2 CO, N-R 24 S0 2

NR

25

CONR

2 6 or Y may be absent; R, R 9

R

1

R

1

R

12

R

1

R

14

R

5

R

16

R

7

R

1

,R'

9

,R

2 0

,R

2

R

22 24 25 26 28 29 3 0

R

3 1

,R

3 2

R

33 3 4 35 36 37

R

38

R

9

R

4 1 ,0R 2 ,0 4

R

44

R

45 and R6are selected from the group consisting of hydrogen, lower alkyl, substituted alkyl, aryl, or substituted aryl;

R

4

R

5

R

47

R

48

R

4 9

R

5 0 and R1are selected from the group consisting of hydrogen, halo, nitro, cyano and -2; U is selected from the group consisting of sulfur, oxygen, NR 28 CO, so, S02, CO 2

NR

29 00 2

NR

30

CONR

3 1

NR

32 S0 2

N

3 3 s0 2

\W

34 S0 2

NR

3 5

NR

3 6 C0, CONR 3 7 p0 2

R

3 8 and P0 3

R

3 9 or U is absent; R R 5 may join together to form a ring; R R and R 3 are selected from the group consisting of hydrogen, alkyl, alkoxycarbonyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aralkyl, cycloalkyl, aryl, substituted aryl, heterocyclo, substituted heterocyclo, cyano, carboxy, carbamyl (e.g.

CONH

2 or substituted carbamyl further selected from CONH alkyl, CONH aryl, CONH aralkyl or cases where there are two substituents on the nitrogen selected from alkyl, aryl or aralkyl;

R

8 and R7are selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aralkyl, cycloalkyl, aryl, substituted aryl, heterocyclo, substituted heterocyclo; Any two of R 1

R

2 and R 3 may join to form a cycloalkyl group; R, S and T are selected from the group consisting of CR 4 0

R

4 1

NR

4 2

R

4 R0is selected from NR 44

R

45

OR

4 6

ON;

G is selected from the group consisting of -S02H, 47 -R48

OH

SOHSH -C -N-

-NHSO

2 OH 0

SR

4 R R51 O N O -N-C-N

I

OH "N 'OH N HOH and heterocycles other than imidazole; A, B, C and D are selected from carbon, oxygen, sulfur or nitrogen, or D may be absent.

with the provisos that 1. R 2 7 may be hydrogen except when U is SO, SO2, C0 2

NR

2 9 CO2 or NR 3 2 SO2, or 2. R 8 may be hydrogen except when Z is SO 2 C0 2 or 0O NR 20

R

21 N NR22 -N*S0 2 -s or 3. Only one of Y, R, S and T may be nitrogen or 4. Any of Y, R, S, and T may be nitrogen except when G is O O R49 II II -N-C -N I I I NHSO2-, OH OH or 15 5. R may be hydrogen except when G is NHSO 2 or 2 6. G may be imidazole only if X is oxygen or NR 7 or 7. Three of r, s, and t together may not be zero.

Listed below are definitions of various terms used to describe 20 this invention. These definitions apply to the terms as they are used throughout this specification, unless otherwise limited in specific instances, either individually or as part of a larger group.

4a Throughout the description and claims of this specification, the word "comprise" and variations of the word, such as "comprising" and "comprises", is not intended to exclude other additives, components, integers or steps.

The term "alkyl" refers to straight or branched chain unsubstituted hydrocarbon groups of 1 to 20 carbon atoms, preferably 1 to 7 carbon atoms. The expression "lower alkyl" refers to unsubstituted alkyl groups of 1 to 4 carbon atoms.

The term "substituted alkyl" refers to an alkyl group substituted by, for example, one to four substituents, such as, halo, trifluoromethyl,

S

o*

S

Se

S

**ee W:\SPECI\94886-98.doc WO 99/18951 PCT/US98/19315 trifluoromethoxy, hydroxy, alkoxy, cycloalkoxy, heterocyclooxy, oxo, alkanoyl, aryloxy, alkanoyloxy, amino, alkylamino, arylamino, aralkylamino, cycloalkylamino, heterocycloamino, disubstituted amines in which the 2 amino substituents are selected from alkyl, aryl or aralkyl; alkanoylamino, aroylamino, aralkanoylamino, substituted alkanoylamino, substituted arylamino, substituted aralkanoylamino, thiol, alkylthio, arylthio, aralkylthio, cycloalkylthio, heterocyclothio, alkyithiono, aryithiono, aralkythiono, alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, sulfonamido, e.g. SO 2

NH

2 substituted sulfonamido, nitro, cyano, carboxy, carbamyl, e.g. CONH 2 substituted carbamyl e.g.

CONH alkyl, CONH aryl, CONH aralkyl or cases where there are two substituents on the nitrogen selected from alkyl, aryl or aralkyl; alkoxycarbonyl, aryl, substituted aryl, guanidino and heterocyclos, such as, indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyl and the like. Where noted above where the substituent is further substituted it will be with halogen, alkyl, alkoxy, aryl or aralkyl.

The term "halogen" or "halo" refers to fluorine, chlorine, bromine and iodine.

The term "aryl" refers to monocyclic or bicyclic aromatic hydrocarbon groups having 6 to 12 carbon atoms in the ring portion, such as phenyl, naphthyl, biphenyl and diphenyl groups, each of which may be substituted.

The term "aralkyl" refers to an aryl group bonded directly through an alkyl group, such as benzyl.

The term "substituted aryl" refers to an aryl group substituted by, for example, one to four substituents such as alkyl, substituted alkyl, halo, trifluoromethoxy, trifluoromethyl, hydroxy, alkoxy, cycloalkyloxy, heterocyclooxy, alkanoyl, alkanoyloxy, amino, alkylamino, aralkylamino, cycloalkylamino, heterocycloamino, dialkylamino, alkanoylamino, thiol, alkylthio, cycloalkylthio, heterocyclothio, ureido, nitro, cyano, carboxy, carboxyalkyl, carbamyl, alkoxycarbonyl, alkyithiono, arylthiono, alkysulfonyl, sulfonamido, aryloxy and the like.

The substituent may be further substituted by halo, hydroxy, alkyl, alkoxy, aryl, substituted aryl, substituted alkyl or aralkyl.

The term "alkenyl" refers to straight or branched chain hydrocarbon groups of 2 to 20 carbon atoms, preferably 2 to 15 carbon WO 99/18951 PCT/US98/19315 6 atoms, and most preferably 2 to 8 carbon atoms, having one to four double bonds.

The term "substituted alkenyl" refers to an alkenyl group substituted by, for example, one to two substituents, such as, halo, hydroxy, alkoxy, alkanoyl, alkanoyloxy, amino, alkylamino, dialkylamino, alkanoylamino, thiol, alkylthio, alkylthiono, alkylsulfonyl, sulfonamido, nitro, cyano, carboxy, carbamyl, substituted carbamyl, guanidino and heterocyclo, e.g. indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyl and the like.

The term "alkynyl" refers to straight or branched chain hydrocarbon groups of 2 to 20 carbon atoms, preferably 2 to 15 carbon atoms, and most preferably 2 to 8 carbon atoms, having one to four triple bonds.

The term "substituted alkynyl" refers to an alkynyl group substituted by, for example, a substituent, such as, halo, hydroxy, alkoxy, alkanoyl, alkanoyloxy, amino, alkylamino, dialkylamino, alkanoylamino, thiol, alkylthio, alkylthiono, alkylsulfonyl, sulfonamido, nitro, cyano, carboxy, carbamyl, substituted carbamyl, guanidino and heterocyclo, e.g. imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyl and the like.

The term "cycloalkyl" refers to a optionally substituted, saturated cyclic hydrocarbon ring systems, preferably containing 1 to 3 rings and 3 to 7 carbons per ring which may be further fused with an unsaturated C3-C7 carbocylic ring. Exemplary groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cycloctyl, cyclodecyl, cyclododecyl, and adamantyl. Exemplary substituents include one or more alkyl groups as described above, or one or more groups described above as alkyl substituents.

The terms "heterocycle", "heterocyclic" and "heterocyclo" refer to an optionally substituted, fully saturated or unsaturated, aromatic or nonaromatic cyclic group, for example, which is a 4 to 7 membered monocyclic, 7 to 11 membered bicyclic, or 10 to 15 membered tricyclic ring system, which has at least one heteroatom in at least one carbon atom-containing ring. Each ring of the heterocyclic group containing a heteroatom may have 1, 2, 3, or 4 heteroatoms selected from nitrogen, oxygen and sulfur, where the nitrogen and sulfur heteroatoms may also optionally be oxidized and the nitrogen heteroatoms may also optionally WO 99/18951 PCTIUS98/1 9315 7 be quaternized. The heterocyclic group may be attached at any heteroatom or carbon atom.

Exemplary monocyclic heterocyclic groups include pyrrolidinyl, pyrrolyl, indolyl, pyrazolyl, oxetanyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, furyl, tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2oxazepinyl, azepinyl, 4-piperidonyl, pyridyl, N-oxo-pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, tetrahydrothiopyranyl, tetrahydropyranyl, morpholinyl, thiamorpholinyl, thiamorpholinyl sulfoxide, tetrahydrothiopyranylsulfone, thiamorpholinyl sulfone, 1,3-dioxolane and tetrahydro-1, 1-dioxothienyl, dioxanyl, isothiazolidinyl, thietanyl, thiiranyl, triazinyl, and triazolyl, and the like. Also included are smaller heterocyclos, such as, epoxides and aziridines.

Exemplary bicyclic heterocyclic groups include benzothiazolyl, benzoxazolyl, benzothienyl, quinuclidinyl, quinolinyl, quinolinyl-Noxide, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuryl, chromonyl, coumarinyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, furopyridinyl (such as furo[2,3-c] pyridinyl, furo[3,1 -blpyridinyll or furo12 ,3-blpyridinyl), dihydroisoindolyl, dihydroquinazolinyl (such as 3,4-dihydro-4-oxoquinazolinyl), benzisothiazolyl, benzisoxazolyl, benzodiazinyl, benzofurazanyl, benzothiopyranyl, benzotriazolyl, benzpyrazolyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone, dihydrobenzopyranyl, indolinyl, isochromanyl, isoindolinyl, naphthyridinyl, phthalazinyl, piperonyl, purinyl, pyridopyridyl, quinazolinyl, tetrahydroquinolinyl, thienofuryl, thienopyridyl, thienothienyl, and the like.

Exemplary substituents include one or more alkyl groups as described above or one or more groups described above as alkyl substituents.

The term "heteroatoms" shall include oxygen, sulfur and nitrogen.

The "ABCD" fused ring to the diazepine ring may be monocyclic or bicyclic, e.g. napthyl or quinolyl in nature.

WO 99/18951 PCT/US98/19315 8 The compounds of formulas I-II may form salts which are also within the scope of this invention. Pharmaceutically acceptable (i.e.

non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful, in isolating or purifying the compounds of this invention.

The compounds of formulas I-II may form salts with alkali metals such as sodium, potassium and lithium, with alkaline earth metals such as calcium and magnesium, with organic bases such as dicyclohexylamine, tributylamine, pyridine and amino acids such as arginine, lysine and the like. Such salts may be obtained, for example, by exchanging the carboxylic acid protons, if they contain a carboxylic acid, in compounds I-II with the desired ion in a medium in which the salt precipitates or in an aqueous medium followed by evaporation.

Other salts can be formed as known to those skilled in the art.

The compounds for formulas I-II may form salts with a variety of organic and inorganic acids. Such salts include those formed with hydrogen chloride, hydroxy methane sulfonic acid, hydrogen bromide, methanesulfonic acid, sulfuric acid, acetic acid, trifluoroacetic acid, maleic acid, benzenesulfonic acid, toluenesulfonic acid and various others nitrates, phosphates, borates, tartrates, citrates, succinates, benzoates, ascorbates, salicylates and the like). Such salts may be formed by reacting compounds I-II in an equivalent amount of the acid in a medium in which the salt precipitates or in an aqueous medium followed by evaporation.

In addition, zwitterions ("inner salts") may be formed.

Compounds of the formulas I-II may also have prodrug forms.

Any compound that will be converted in vivo to provide the bioactive agent the compound for formulas I-II) is a prodrug within the scope and spirit of the invention.

For example compounds of the formulas I-II may be a carboxylate ester moiety. The carboxylate ester may be conveniently formed by esterifying any of the carboxylic acid functionalities found on the disclosed ring structure(s).

Various forms of prodrugs are well known in the art. For examples of such prodrug derivatives, see: WO 99/18951 PCT/US98/19315 9 a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) and Methods in Enzvmology, Vol.42, p. 309-396, edited by K. Widder, et al. (Acamedic Press, 1985); b) A Textbook of Drug Design and Development, edited by Krosgaard-Larsen and H. Bundgaard, Chapter 5, "Design and Application of Prodrugs," by H. Bundgaard, p. 113-191 (1991); c) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992); d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285 (1988); and e) N. Kakeya, et al., Chem Phar Bull, 32, 692 (1984).

It should further be understood that solvates hydrates) of the compounds of formulas I-IV are also with the scope of the present invention. Methods of solvation are generally known in the art.

Preferred Moieties For compounds of the present invention, the following moieties are preferred: Compounds of formulas I, and II wherein m is zero.

More preferred are compounds of Formula I, and II wherein m is zero and X is carbon.

Most preferred are compounds of formula I wherein m, is zero, X is methylene (CH 2 and "ABCD" is a carbocyclic ring, benzo.

Use and Utility The compounds of formulas I-II are inhibitors of S-farnesyl protein transferase. They are thus useful in the treatment of a variety of cancers, including (but not limited to) the following; carcinoma, including that of the bladder, breast, colon, kidney, liver, lung, including small cell lung cancer, ovary, prostate, testes, pancreas, esophagus, stomach, gall bladder, cervix, thyroid and skin, including squamous cell carcinoma; hematopoietic tumors of lymphoid lineage, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma, and Burketts lymphoma; WO 99/18951 PCTIUS98/1 9315 hematopoietic tumors of myeloid lineage, including acute and chronic myelogenous leukemias, myelodysplastic syndrome and promyelocytic leukemia; tumors of the central and peripheral nervous system, including astrocytoma, neuroblastoma, glioma, and schwannomas; tumors of mesenchymal origin, including fibrosarcoma, rhabdomyoscarcoma, and osteosarcoma; other tumors, including melanoma, xenoderma pigmentosum, keratoactanthoma, seminoma, thyroid follicular cancer and teratocarcinoma.

The compounds of formulas I-II are especially useful in treatment of tumors having a high incidence of ras involvement, such as colon, lung, and pancreatic tumors and in tumors in which a prenyl transferase contributes to tumor maintenance, tumor growth or tumor development. By the administration of a composition having one (or a combination) of the compounds of this invention, development of tumors in a mammalian host is reduced, or tumor burden is reduced, or tumor regression is produced.

Compounds of formulas I-II may also inhibit tumor angiogenesis, thereby affecting the growth of tumors. Such antiangiogenesis properties of the compounds of formulas I-II may also be useful in the treatment of certain forms of blindness related to retinal vascularization.

Compounds of formulas I-II may also be useful in the treatment of diseases other than cancer that may be associated with signal transduction pathways operating through ras, e.g., neurofibromatosis, atherosclerosis, pulmonary fibrosis, arthritis, psoriasis, glomerulonephritis, restenosis following angioplasty or vascular surgery, hypertrophic scar formation, polycystic kidney disease and endotoxic shock. Compounds I-II may be useful as anti-fungal agents.

Compounds of formula I-II may induce or inhibit apoptosis, a physiological cell death process critical for normal development and homeostasis. Alterations of apoptotic pathways contribute to the pathogenesis of a variety of human diseases. Compounds of formula I- II, as modulators of apoptosis, will be useful in the treatment of a variety of human diseases with aberrations in apoptosis including cancer W/ o01 Qo C PCT/ SQ9T/1 9315 I I (particularly, but not limited to follicular lymphomas, carcinomas with p53 mutations, hormone dependent tumors of the breast, prostrate and ovary, and precancerous lesions such as familial adenomatous polyposis), viral infections (including but not limited to herpes virus, pox virus, Epstein-Barr virus, Sindbis virus and adenovirus), autoimmune diseases (including but not limited to systemic lupus erythematosus, immune mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowl diseases and autoimmune diabetes mellitus), neurodegenerative disorders (including but not limited to Alzheimer's disease, AIDS-related dementia, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentosa, spinal muscular atrophy and cerebellar degeneration), AIDS, myelodysplastic syndromes, aplastic anemia, ischemic injury associated myocardial infarctions, stroke and reperfusion injury, arrhythmia, atherosclerosis, toxin-induced or alcohol induced liver diseases, hematological diseases (including but not limited to chronic anemia and aplastic anemia), degenerative diseases of the musculoskeletal system (including but not limited to osteoporosis and arthritis), aspirin-sensitive rhinosinusitis, cystic fibrosis, multiple sclerosis, kidney diseases, and cancer pain.

Compounds of formulas I-II may also be useful in the treatment of diseases associated with farnesyl transferase substrates other than ras nuclear lamins, transducin, rhodopsin kinase, cGMP phosphodiesterase, TC21, phosphorylase kinase, Rap2, RhoB, RhoE, PRL1) that are also post-translationally modified by the enzyme farnesyl protein transferase.

Compounds of formulas I-II may also act as inhibitors of other prenyl transferases geranylgeranyl transferase I and II), and thus be effective in the treatment of diseases associated with other prenyl modifications geranylgeranylation) of proteins the rap, rab, rac and rho gene products and the like). For example, they may find use as drugs against Hepatitis delta virus (HDV) infections, as suggested by the recent finding that geranylgeranylation of the large isoform of the delta antigen of HDV is a requirement for productive viral infection S.

Glen, et al., Science, 256, 1331 (1992)].

The compounds of this invention may also be useful in combination with known anti-cancer and cytotoxic agents and treatments, including radiation. If formulated as a fixed dose, such W 00/1 90I PrT/i ICOQ/1i 011 12 combination products employ the compounds of this invention within the dosage range described below and the other pharmaceutically active agent within its approved dosage range. Compounds of formulas I-II may be used sequentially with known anticancer or cytotoxic agents and treatment, including radiation when a combination formulation is inappropriate.

Farnesyl transferase assays were performed as described in V.

Manne et al., Drug Development Research, 34, 121-137, (1995). The compounds of Examples 1-17 inhibited farnesyl transferase with IC values between 0.lnM and 100uM.

The compounds of this invention may be formulated with a pharmaceutical vehicle or diluent for oral, intravenous, intraperitoneal, subcutaneous, intraabdominal, intramuscular, rectal, vaginal or topical administration. Oral administration may involve the use of slow release formulations, such as biodegradable polymers or prodrugs. The pharmaceutical composition can be formulated in a classical manner using solid or liquid vehicles, diluents and additives appropriate to the desired mode of administration. Orally, the compounds can be administered in the form of tablets, capsules, granules, powders and the like. The compounds may be administered in a dosage range of about 0.05 to 200 mg/kg/day, preferably less than 100 mg/kg/day, in a single dose or in 2 to 4 divided doses.

WO 99/1 8951 PTU9I191 PCTIUS98/19315 Process of Prelparation Scheme 1

NH

2 R 0 HCI- 2 N ~'CO 2 Et Step 1 k\ Sep R, 0

NH

A ~-R2 Step 3 K LN 7R

H

4 R1 Zj-R 3 N2 Step 4

R

4

CHO

Step 5

IR

4 00X Step 6 R N '1R3

N

R

4 '1 0 7 wherein R, is selected from H, halo, NO 2

NH

2 CN, alkyl, aryl, heteroaryl, substituted alkyl, arylalkyl, alkoxy and substituted amino;

R

2

R

3 and R 4 are selected from alkyl, substituted alkyl, arylalkyl, aryl, heteroaryl; R 2 can also be hydrogen; Z is selected from GO, SO 2 Go 2 CONR 5 ,S S 2

NR'.

Sfte The isatoic anhydride 2 was formed by reaction of optionally substituted anthranilic acid with a phosgene equivalent, such as, phosgene or triphosgene in a mixed aqueous/organic solvent at from 0 0

C

to 50'C range.

wn o/1 n951 PCT/US98/19315 14 The isatoic anhydride 2 is reacted with an amino acid ester hydrochloride salt in pyridine at an elevated temperature with reflux as preferred to give benzodiazepinedione 3. The compound 3 wherein R1 is aryl or heteroaryl can be prepared from the compound 3 wherein R1 is bromo by a palladium coupling of an aryl or heteroaryl metaloid derivative such as phenylboronic acid in a mixed aqueous/organic solvent, e.g. THF/DMF/water, in the presence of a base, e.g. sodium carbonate, at from room temperature to 110°C.

Step 3 Thereafter the compound 3 is reacted with a reducing agent such as lithium aluminum hydride or borane in an inert atmosphere and in an organic solvent, such as, tetrahydrofuran at from room temperature to reflux.

Stp4 Thereafter the product is acylated or sulfonylated under standard conditions at from -78 0 C to 100 0 C by reaction with an acid halide R3COX wherein X is Cl or Br in an inert organic solvent, e.g.

acetonitrile, or in a mixed aqueous/organic solvent e.g.

NaOH/dichloroethane; by reaction with a sulfonyl halide R3SO2X wherein X is Cl or Br in an organic solvent such as THF in the presence of a base such as diisopropylethylamine or in a mixed aqueous/organic solvent e.g. NaOH/CH2C12). Where the compound 5 is sulfonylated with a beta-haloalkylsulfonyl halide, the halide may then be eliminated by a base such as diisopropylethylamine and then nucleophiles such as dimethylamine may be added to the resulting unsaturated sulfonamide by treatment in an organic solvent such as THF or dichloromethane at from room temperature to reflux.

Thereafter the various products can undergo reductive alkylation with an aldehyde in the presence of an acid e.g. acetic acid, a reducing agent e.g. NaBH(OAc)3 in an inert organic solvent e.g.

dichloroethane at about room temperature to form the compounds of formula I and II. The aldehydes may be prepared from the appropriate WO 99/18951 PCT/US98/19315 alcohols by oxidation with for example oxalyl chloride/DMSO/triethylamine. They may be prepared from appropriate carboxylic acid derivatives such as esters by reduction with for example diisobutylaluminum hydride in an organic solvent such as methylene chloride at from -78C to room temperature.

Compound 6 may be further manipulated to give the compounds of formula I and II, such as by reaction with a deprotecting agent like TFA/triethylsilane in an organic solvent such as dichloroethane.

If the compound 6 contains a leaving group such as chloride; in the case of a hydroxyl group, such a group may be transformed into a leaving group such as trifluoromethanesulfonate; the compound may be reacted with a nucleaphile such as 2-aminopyridine to give the desired compounds of formula I or II.

The compound 5 is acylated with an acylating agent such as an acid chloride in a solvent such as methylene chloride in the presence of an optional base such as diisopropylethylamine. The compound 5 may also be acylated with an isocyanate which in turn can be prepared by an acyl azide like nicotinoyl azide. If R 4 contains a leaving group such as chloride, the compound may be further reacted with a nucleophile such as 2-pyrrolidinone or ammonia to form the compounds of formula I or

II.

WO 99/18951 PCT/US98/19315 Scheme 2

-R

NCO

CO

2 Et

,R

Z-R3 N Z

R

Step 2 HOH N2H O D2H 0 Step 1 Step 3

RN'

R NZI-R3 N R

CHO

Step 4 R, NZrR3 Step

HO

N 0 R4

HNB

n OBn 4 5 0 wherein RI is selected from H, halo, NO 2

NH

2 CN, alkyl, aryl, heteroaryl, substituted alkyl, arylalkyl, alkoxy and substituted amino;

R

2

R

3 and R 4 are selected from alkyl, arylalkyl, aryl, heteroaryl; R 2 and

R

4 can also be hydrogen; R 4 can also be amino; Z is selected from CO, SO,, CO 2

CONR

5

SO

2

NR

5 Stepl An ester 1 prepared by following Step 5 of Scheme 1 may be saponified to an acid 2 with a base such as LiOH in a solvent such as aq.

methanol in THF at about room temperature.

Thereafter the acid 2 is converted to hydroxamic acid 3 with hydroxylamine hydrochloride in the presence of a dehydrating agent such as 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and a base such as triethylamine.

wn 9/18951 PCT/US98/19315 17 The ester 1 may be reduced to an aldehyde 4 by a reducing agent such as diisobutylaluminium hydride in an organic solvent such as methylene chloride.

Step4 Thereafter, the aldehyde 4 may be reductively aminated with a hydroxylamine derivative such as O-benzyl hydroxylamine and a reducing agent such as sodium cyanoborohydride to give The compound 5 may be acylated with an acylating agent such as formic anhydride to give after deprotection, e. by hydrogenation the compound 6.

The invention will now be further described by the following working examples, which are preferred embodiments of the invention.

All temperatures are in degrees Celsius (oC) unless otherwise indicated.

These examples are illustrative rather than limiting.

Example 1 0 (+,-)-1-(2-Aino-3-mercaptopropyl)-2,3,4,5-tetrahydro4- 2 5 (naphthalenylcarbonyl)-1H-1,4-benzodiazepine, monohydrochloride A. 2,3,4,5-Tetrahydro-1H-benzodiazepine-2,5-dione.

A stirred solution of isatoic anhydride (16.4 g, 0.1 mol) and glycine ethyl ester hydrochloride in 40 mL of pyridine was heated under reflux on an oil-bath for 7 h. The suspension was cooled to 0°C for 18 h. The precipitate was collected, and washed with ethanol and ether to give a light yellow solid (7.0 g, WO 99/18951 PCTIUS98/1 9315 18 B. 2,3,4,5-Tetrahydro-1H-1,4-benzodiazepine To a stirred suspension of LAH (3.5 g, 90 mmol) in THF (100 mL) at room temperature was slowly added compound A (3.5 g, 20 mmol) portionwise at room temperature under argon. The resultant suspension was heated on an oil-bath at reflux under argon for 18 h and cooled to 0 C. A solution of ammonium hydroxide (5 mL, cone.) in 30 mL of THF was added via an additional funnel. The resultant suspension was stirred for 1 h and filtered. The filtrate was concentrated in vacuo to give an oil (2.5 g, 84%).

C. 2,3,4,5-Tetrahydro-4-(naphthalenylcarbonyl)-1H-1,4benzodiazepine To a stirred solution of the compound B (2.0 g, 13.4 mmol) in a mixed solvent of CH2Cl2/1N NaOH 150 mL), a solution of 1naphthoyl chloride (2.0 ml, 13.4 mmol) in CH2C12 (3 mL) was added slowly at 0°C. The biphasic mixture was allowed to stir at 0°C for 2 hr.

The aqueous phase was extracted with CH2C12 (2x100 mL) The combined organic layers were dried over Na2SO4, concentrated to provide the title compound (3.6 g, 89%) as a white solid. MS (M H) 303.

Anal. cal'd for C 20

H

8 N20: C, 89.44; H, 6.00; N, 9.26.

Found: C, 89.48; H, 5.98; N, 9.14.

D. (+,-)-1-(2-Amino-3-mercaptopropyl)-2,3,4,5-tetrahydro-4- (naphthalenylcarbonyl)-1H-1,4-benzodiazepine, monohydrochloride To a stirred solution of compound C (102 mg, 0.34 mmol) and N- BOC-S-trityl-cysteine aldehyde (170 mg, 0.38 mmol) in a mixture of dichloroethane (5 mL) and acetic acid (0.5 mL), was added NaBH(OAc) 3 (150 mg, 0.68 mmol) in one portion. The reaction was allowed to proceed for 30 min, the mixture was diluted with ethyl acetate (50 mL), followed WO 99/18951 PCT/US98/19315 19 by addition of sat. NaHCO 3 (5 mL). The resultant mixture was stirred for 2 h, and poured into a mixture of ethyl acetate and saturated NaHCO, and well shaken. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (50 mL). The combined organic extracts were dried over MgSO 4 and concentrated in vacuo. The residue was purified by flash column chromatography (1:2 ethyl acetate and hexanes) to give an oil 220 mg, This was dissolved in a solvent of TFA and methylene chloride (5 mL, The resultant solution was stirred at room temperature for 3 h. The solvent was removed in vacuo and the residue was partitioned in ether (10 mL) and 10% HC1 solution mL). The aqueous solution was separated, and concentrated in vacuo to a small volume. The product was purified by preparative HPLC (aqueous methanol in the presence of 0.1%TFA) and obtained as HC1 salt after salt exchange (73 mg, 62%, mp: 140°C). MS 392.

Example 2

NH,

0 (+,-)-1-(2-Amino-3-mercaptopropyl)-2,3,4,5-tetrahydro4- (naphthalenylcarbonyl)-7-phenyl-lH-l,4-benzodiazepine, monohydrochloride A. 5-Bromoisatoic Anhydride To a stirred solution of 5-bromo-2-amino-benzoic acid (25 g, 0.115 mmol) in a mixed solvent of THF:H20:conc. HC1 350 mL), was added a solution of phosgene (0.23 mmol, 20% in toluene) dropwise. The reaction mixture was allowed to stir at room temperature for 4 h. The wn 99/I8951 PCT/US98/19315 resulting precipitate was filtered, washed with water and dried under reduced pressure to give the title compound (22.5 g, 78%) as a gray solid.

B. 2,3,4,5-Tetrahydro-7-bromo-1H-1,4-benzodiazepine-2,5-dione The title compound was prepared by following the procedure for the preparation of compound A of Example 1.

C. 2,3,4,5-Tetrahydro-7-phenyl-lH-1,4-benzodiazepine-2,5-dione To a stirred, degassed solution of compound B (13.0 g, 51 mmol) in a solvent of DMF/THF 250 mL), tetrakis(triphenyl phosphine) palladium(0) (10.5 g, 9.1 mmol) was added. After stirring for 0.5 hr, an aqueous solution of Na2CO3 (8.5 g, 80 mmol) was added followed by benzeneboronic acid (10.2 g, 83 mmol) The resultant suspension was heated at 105 OC for 24 hrs and filtered. The filtrate was concentrated and diluted with H20 (200 mL). The precipitate was filtered, and washed with H20 (200 mL), CH2C12 (50 mL) and ethyl acetate (100 mL) to afford the title compound as a yellow solid (10.8 g, MS 253.

D. 2,3,4,5-Tetrahydro-7-phenyl-1H-1,4-benzodiazepine The title compound was prepared by following the procedure for the preparation of compound B of Example 1. MS 225.

E. 2,3,4,5-Tetrahydro-4-(naphthalenylcarbonyl)-7-phenyl-1H-1,4benzodiazepine The title compound was prepared by following the procedure for the preparation of compound C of Example 1. MS 379.

WO 99/18951 PCT/US98/19315 WO 99/18951 PTU9/91 21 F. (+,-)-1-(2-Amino-3-mercaptopropyl)-2,3,4,5-tetrahydro4- (naphthalenylcarbonyl)-7-phenyl-1H-1,4-benzodiazepine, monohydrochloride The title compound was prepared by following the procedure for the preparation of compound D of Example 1. MS 468.

Example 3 100 H O 2,3,4,5-Tetrahydro-4-(1-naphthalenylsulfonyl)-N-(3-pyridinyl)-1H-1,4benzodiazepine-1-carboxamide A. 2,3,4,5-Tetrahydro-4-(1-naphthalenylsulfonyl)-1H-1,4benzodiazepine To a stirred solution of the compound B of Example 1 (0.75 g, mmol) in methylene chloride and 1 N NaOH solution (30 mL, 3:1) at 0 0

C,

was added 1-naphthalenesulfonyl chloride (1.13 g, 5.0 mmol). The mixture was allowed to stir at o'C for 30 min and poured into a mixture of methylene chloride and 1 NaOH solution (50 mL each). The organic layer was separated and washed with NH 4 C1 solution, dried over MgSO, and concentrated in vacuo. The residue was crystallized from methanol to give a solid (1.3 g, 78%, mp: 165-166 0 MS 339.

B. 2,3,4,5-Tetrahydro-4-(l-naphthalenylsulfonyl)-N-(3-pyridinyl)-1H- 1,4-benzodiazepine-1-carboxamide A stirred solution of the compound A (160 mg, 0.47 mmol), and 3nicotinyl azide (90 mg, 0.61 mmol) in toluene (2 mL) was heated at 100 0

C

PCT/US98/19315 WO) 99/18951PCUS/195 22 for 3 h. The mixture was cooled to room temperature. The solvent was removed in vacuo and the residue was crystallized from ether to give a white solid (150 mg, MS 459.

Anal. calc'd C 25

H

22

N

4 0 3 S *0.30H 2 0: C, 64.72; H, 4.91; N, 12.08; S, 6.91.

Found: C, 64.63; H, 4.89; N, 11.86; S, 6.54.

Example 4 Br 0 o1 0 000 (R)-7-Bromo-2,3,4,5-tetrahydro4-(methylsulfonyl)--I(2-oxo-1.

pyrolidinyl)acetyl-3-(phenylmethyl)-lH-1,4-benzodiazepine, trifluoroacetate.

A. (R)-2,3,4,5-Tetrahydro-7-bromo-3-phenylmethyl-benzodiazapine- A stirred solution of the compound A of Example 2 (150 g, 0.62 mol) and D-phenyl alanine methyl ester hydrochloride (127.3 g, 0.59 mol) in the presence of 4-dimethylaminopyridine (2 g) in pyridine (1500 mL) was heated at reflux under argon for 3 days. The pyridine was removed in vacuo and the residue was dissolved in methylene chloride (3 The solution was washed with 10% HCl solution and brine, dried, and concentrated in vacuo to a small volume. The solid thus formed was collected and dried to give 152 g of solid (mp: 242-243'C; 71%).

B. (R)-2,3,4,5-Tetrahydro-7-bromo-3-phenylmethyl-benzodiazapine To a stirred solution of compound A (30 g, 87 mmol) in anhydrous THF (870 mL) under argon, was added a solution of borane wn ooI l989 PCT/ISq/19315i 23 tetrahydrofuran complex (440 mL of a 1 M solution, 440 mmol) at room temperature. The solution was slowly heated to reflux, and maintained at reflux for 18 h. The mixture was cooled to 0°C, and methanol (150 mL) was added slowly. The resultant solution was concentrated in vacuo.

The residue was dissolved in methanol (250 mL) and 7 N HC1 solution mL) was added. The mixture was heated on a steam-bath for 2 h. The solid thus formed was collected, resuspended in water (400 mL) and the aqueous suspension was made basic to pH 11 with 5 N NaOH solution and extracted with ethyl acetate 2 x 300 mL). The organic extracts were combined, dried, concentrated in vacuo and the residue was crystallized from methanol and water to give a white solid (mp: 135-138 0 C, 25 g, 91%).

C. (R)-2,3,4,5-Tetrahydro-7-bromo-4-methanesulfonyl-3phenylmethyl-benzodiazapine To a stirred solution of compound B (1.5 g, 4.73 mmol), pyridine (3 mL), and diisopropylethyl amine (1.6 mL, 9.46 mmol) in methylene chloride, was added methanesulfonyl chloride (0.55 mL, 7.11 mmol) at 0°C under argon. The resultant mixture was allowed to stir at 0 C for 2 h, 1 N NaOH solution (30 mL) was added and the mixture was stirred for 2 h. The organic layer was separated, washed with 1 N HC1 solution (2 x 100 mL), dried, and concentrated in vacuo to give a yellow solid (1.7 g, 91 D. (R)-7-Bromo-2,3,4,5-tetrahydro-4-(methylsulfonyl)-l-chloroacetyl-3- (phenylmethyl)-lH-1,4-benzodiazepine To a stirred solution of compound C (2.0 g, 5.06 mmol) and diisopropylethylamine (4.4 mL, 25 mmol) in dichloromethane (100 mL) in an ice bath under argon, was added chloroacetyl chloride (2.0 mL, 25.3 mmol). The mixture was allowed to stir for 30 min., poured into 1N aqueous sodium hydroxide solution (200 mL) and extracted with dichloromethane (2 x 100 mL). The organic extracts were combined, WO 09/1R951 PCT/US98/Z19315 24 washed with 10% HCI solution and brine, dried (MgSO 4 and concentrated in vacuo to an oil, which was purified by chromatography (3 1, hexane ethylacetate) to give the title compound as a colorless oil (760 mg, 33 E. (R)-7-Bromo-2,3,4,5-tetrahydro-4-(methylsulfonyl)--[(2-oxo-1pyrrolidinyl)acetyl]-3-(phenyhnmethyl)-1H-1,4-benzodiazepine, trifluoroacetate.

To a stirred mixture of compound D (50 mg, 0.11 mmol) and 2pyrrlidinone (0.25 mL, 0.33 mmol) in dimethylformamide, was added sodium hydride (14 mg, 0.33 mm) at room temperature under argon.

The mixture was allowed to stir at room temperature for 2 h, and poured into a mixture of 1N aqueous hydrochloric acid (75 mL), and ethyl acetate (75 mL). The organic layer was separated, dried (Na 2

SO

4 and concentrated in vacuo. The residue was purified by preparative HPLC (aqueous methanol containing 0.1% trifluoroacetic acid). The product fractions were combined and lyophilized to provide the title compound as a white solid (15 mg, 26 MS (M 521.

Example Br OZ: N I N 0,

H

2

N

(R)-7-Bromo-4-[[2-(dimethylamino)ethylsulfonyl]-2,3,4,5-tetrahydro-boxo- 3 -(phenyhnlmethyl)-1H-1,4-benzodiaze-pine-l-ethanesulfonamide.

wn 99/1i8 PCT/IIS98/1931 A. (R)-7-Bromo-2,3,4,5-tetrahydro-4-(ethenylsulfonyl)-3- (phenylmethyl)-1H-l,4-benzodiazepine To a stirred solution of the compound B of Example 4 (10 g, 31.5 mmol) in dichloromethane (120 mL) at 0°C, was added dropwise a solution of 2-chloroethanesulfonyl chloride (3.2 mL, 30 mmol) in dichloromethane (10 mL) followed by dropwise addition of DIPEA (5.2 mL, 30 mmol). After stirring for 15 min., 2-chloroethanesulfonyl chloride (1.5 mL, 15 mmol) followed by DIPEA (10.4 mL, 60 mmol) were added. The mixture was allowed to warm to room temperature and poured into water (80 mL). The organic layer was separated, washed with 1N HC1 and saturated aqueous NaHCO 3 (80 mL each), dried (MgSO- 4) and concentrated in vacuo to afford the title compound as a yellowish foamy solid (15.2 MS 406.

B. (R)-7-Bromo-2,3,4,5-tetrahydro-4-[[2-(dimethylamino)ethyl]sulfonyl]-3-(phenylmethyl)-lH-1,4-benzodiazepine A round bottomed flask was charged with compound A (7 g) and a THF solution of dimethylamine (2 M, 20 mL) at room temperature. The flask was stoppered with a glass stopper and the mixture was allowed to stir for 18 hr. The mixture was concentrated and purified by chromatography (20% acetone in chloroform) to afford title compound (3.25 g, 48% from the compound B of Example MS 451.

C. (R)-7-Bromo-4-[[2-(dimethylamino)ethyl]sulfonyl]-2,3,4,5tetrahydro-b-oxo-3-(phenylmethyl)-lH-1,4-benzodiazepine-1ethanesulfonamide To a stirred solution of compound B (90 mg, 0.2 mmol) in dichloromethane (1 mL) at 0°C under argon, was added chlorosulfonylacetyl chloride (0.063 mL, 0.6 mmol). The mixture was allowed to stir for 8 hr, cooled to -78 0 C and liquid ammonia (2 mL) was condensed into the mixture. The stirred mixture was allowed to warm to WO 99/18951 PCT/IIU98/9315; 26 room temperature over 1 hr, diluted with chloroform (20 mL) and shaken with water (20 mL). The aqueous layer was adjusted to pH 6-7 by careful addition of 1N HC1. The organic layer was separated, dried

(K

2

CO

3 and concentrated. The residue was purified by chromatography (with 10% i-PrOH in chloroform) to afford title compound (51 mg, MS 573.

Example 6 0 2 3 ,4,5-Tetrahydro-4-(-naphthalenylcarbonyl)-l-(2-thienylmethyl)-lH- 1,4-benzodiazepine.

To a stirred solution of the compound C of Example 1 (100 mg, 0.33 mmol), 2-thiophenecarboxaldehyde (56 mg, 0.5 mmol) in a mixture of dichloroethane (2 mL) and acetic acid (1.0 mL), NaBH(OAc) 3 (190 mg) was added in one portion. The mixture was stirred for 30 min, and diluted with ethyl acetate (25 mL), followed by NH 4 OH (3 mL, conc.). The resultant mixture was stirred at room temperature for 18 h and poured into a mixture of ethyl acetate (50 mL) and sat. NaHCO 3 (50 mL). This was well shaken. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (50 mL). The combined organic extracts were dried (MgSO 4 and concentrated in vacuo. The residue thus obtained was purified by chromatography (3:1 hexanes and ethyl acetate) to give a solid (40 mg, MS 399.

WO 99/18951 WO 9918951PCT[US98/1 9315 27 Example 7 2,3,4,5-Tetrahydro-4-(1-naphthalenylcarbonyl)7phenyl1-(3pyridinymethyl)-1H-1,4-benzodiazepine, hydrochloride.

The title compound was prepared from nicotinaldehyde and the compound E of Example 2 by following the procedure described for the preparation of Example 6. MS 470.

Example 8 2 3 ,4,5-Tetrahydro-4-(1-naphthalenylcarbonyl)-l.(3-thienylmethyl)-1H- 1,4-benzodiazepine The title compound was prepared from 3thiophenecarboxaldehyde and the compound C of Example 1 by following the procedure described for the preparation of Example 6. MS 475.

WO 99/18951 PCT/US98/19315 28 Example 9 N

NO

HO O 2,3,4,5-Tetrahydro-1-[2-(3-hydroxy-2-oxo-1(2H)-pyridinyl)ethyl]-4-(1naphthalenylcarbonyl)-1H-1,4-benzodiazepine, dihydrochloride.

A. Ethyl 3-hydroxy-2-oxo-1(2H)-pyridinylacetate A stirred mixture of 2,3-dihydroxypyridine (11.lg, 0.10 mol) and ethyl bromoactetate (55.4 mL, 0.5 mol) was refluxed for 12 h, cooled, and concentrated. The residue was recrystallized from ethyl acetate/hexanes to give the title compound as a white solid (19.5 g).

MS 196.

B. Ethyl 3-[dimethyl-(1,1-dimethyl)ethylsloxy-2-oxo-1(2H)pyridinylacetate To a stirred solution of compound A (1.0 g, 5.1 mmol) and imidazole (863 mg, 12.7 mmol) in dimethylformamide (5 mL), was added tert-butyldimethylchlorosilane (920 mg, 6.1 mmol). The resultant solution was allowed to stir for 1 h, and poured into water (100 mL). The aqueous solution was extracted with ethyl acetate. The organic layer was washed with water, 10% aq. lithium chloride, dried (MgSO 4 filtered and concentrated to give the title compound as a clear oil (1.5 MS 312.

WO 99/18951 PCT/US98/19315 29 C. 3-[Dimethyl-(1,1-dimethyl)ethylsiloxy]-2-oxo-1(2H)pyridinylethanal.

To a stirred solution of compound B (450 mg, 1.44 mmol) in dry methylene chloride (5 mL) at -78 0 C, was added diisobutylaluminum hydride (1M in methylene chloride, 2.2 mL, 2.2 mmol) dropwise over min. The mixture was stirred for 1 h at -78 0 C and allowed to warm to rt.

Water was added and the methylene chloride layer was separated, washed with water, dried (MgSO 4 filtered and concentrated to give title compound as a clear oil (385 mg). MS 268.

D. 2,3,4,5-Tetrahydro-1-[2-(3-hydroxy-2-oxo-1(2H)-pyridinyl)ethyl]-4-(1naphthalenylcarbonyl)-1H-1,4-benzodiazepine, dihydrochloride The title compound was prepared from compound C and the compound C of Example 1 by following the procedure described for the preparation of Example 6. The product was dissolved in methanol, 1 N HC1 in ether was added, and the solvent was removed to give title compound. MS 440.

Example N

N

HO O 2,3,4,5-Tetrahydro-1-[2-(3-hydroxy-2-oxo-1(2H)-pyridinyl)ethyl]4-(1naphthalenylcarbonyl)-7-phenyl-1H-1,4-benzodiazepine, dihydrochloride.

WO 99/18951 PCT/US98/19315 The title compound was prepared from the compound C of the Example 9 and the compound E of Example 2 by following the procedure described for the preparation of Example 6. The product was dissolved in methanol, 1 N HC1 in ether was added, and the solvent was removed to give title compound. MS 516.

Example 11 0N.

o 2,3,4,5-Tetrahydro-4-(-naphthalenylcarbonyl)-l-[(5-oxo-2pyrrolidinyl)carbonyl]-7-phenyl-1H-1,4-benzodiazepine.

To a stirred solution of pyroglutamic acid (400 mg, 3.1 mmol) in methylene chloride (6 mL), was added thionyl chloride (4 mL) along with a drop of pyridine at room temperature. The reaction mixture was allowed to stir for 2 h and concentrated. The residue was dissolved in 2 mL methylene chloride. To this stirred solution, was added the compound E of Example 2 (40 mg, 0.106 mmol) and diisopropylethylamine (0.5 mL, 3 mmol) in methylene chloride (5 mL).

The mixture was allowed to stir for 16 h, and concentrated. The residue was dissolved in ethyl acetate (50 mL). The resultant solution was washed successively with IN aqueous potassium bisulfate, saturated aqueous sodium bicarbonate and brine. The organic layer was dried (MgSO,), filtered and concentrated. The residue was purified by preparative HPLC (aqueous methanol with 0.1% TFA) to give 15 mg of the title compound as a white solid. MS (M+NH 4 507.

WO 99/18951 PCTIUS98/19315 31 Example 12

I-

NN

2,3,4,5-Tetrahydro-4-(1-naphthalenylcarbonyl).7-phenyl-.-[3-(2pyridinylamino)propyl]-1H-1,4-benzodiazepine, dihydrochloride.

A. 3-[Dimethyl(1,1-dimethylethyl)siloxy]-propanol To a stirred solution of 1,3-propanediol (4.8 mL, 66 mmol) in THF (100 mL) was added sodium hydride (60% dispersion in mineral oil, 2.6 g, 66 mmol) followed by tert-butyldimethyl chlorosilane. The solution was allowed to stir for 16 h, and concentrated to an oily residue.

Purification by chromatography (40% ethyl acetate in hexanes) afforded title compound (12.3 MS 191.

B. 3-[Dimethyl(1,1-dimethylethyl)siloxy]-propanal To a stirred solution of oxalyl chloride (504 mg, 4 mmol) in 20 mL methylene chloride at -63°C under argon, was added dimethyl sulfoxide g, 2.6 mmol) in methylene chloride (5 mL). This was followed by addition of the compound A (1.0 g, 2.6 mmol) in methylene chloride mL) over 15 min keeping the reaction temperature below -50°C. The resultant solution was stirred for 30 min at -63°C, then a solution of triethylamine (1.2 g, 11.9 mmol) in methylene chloride (5 mL) was added over 15 min keeping the solution below -50°C. The reaction was allowed to stir for 30 min, and quenched by addition of 1M potassium hydrogen WO 99/18951 PCT/US98/19315 32 sulfate (4.5 mL), water (20 mL) and ethyl ether (200 mL). The layers were separated and the aqueous layer was made basic using saturated aqueous sodium bicarbonate and extracted with ethyl acetate (3 x 50 mL).

The combined organic layers were dried (MgSO4) and concentrated to yield title compound (1.0 MS 189.

C. 2,3,4,5-Tetrahydro-4-(1-naphthalenylcarbonyl)-7-phenyl-1-[3- Dimethyl(1,1-dimethylethyl)siloxypropyl]-lH-1,4-benzodiazepine The title compound was prepared from compound B and the compound E of Example 2 by following the procedure described for the preparation of Example 6. MS 552.

D. 2,3,4,5-Tetrahydro-4-(1-naphthalenylcarbonyl)-7-phenyl-1-[3hydroxypropyl]-1H-1,4-benzodiazepine A solution of compound C (300 mg, 0.54 mmol) in dioxane (6 mL) and 4 N aqueous HC1 solution was stirred for lh and concentrated. The residue was triturated with diethyl ether to give 230 mg of title compound as a white solid. MS 437.

E. 2,3,4,5-Tetrahydro-4-(1-naphthalenylcarbonyl)-7-phenyl-1-[3-(2pyridinylamino)propyl]-1H-1,4-benzodiazepine, dihydrochloride To a stirred solution of compound D (50 mg, 0.11 mmol) in dry methylene chloride (5 mL) at -40 0 C, was added di-(tert-butyl)methylpyridine (23 mg, 0.11 mmol) followed by triflic anhydride (0.19 mL, 0.11 mmol). The solution stirred for 30 min at -40 0 C and 2aminopyridine (16 mg, 0.17 mmol) was added. The mixture was allowed to stir at -40°C for 30 min and warmed to room temperature over 2 h. The reaction was concentrated and purified by preparative HPLC (aqueous methanol with 0.1% TFA). The product was dissolved in methanol, 1 N HC1 in ether was added, and the solvent was removed to give the title compound as a solid (20 mg). MS 513.

WO 99/18951 WO 9918951PCTIUS98/1 9315 33 Example 13 ((7-NH 2,3,4,5-Tetrahycdro-4-(-naphthalenylcarbonyl)-7-phenyl--[3-(3pyridinylamino)propyl-1H-1,4-benzodiazepine, dihydrochioride.

The title compound was prepared by following the procedure described for the preparation of Example 12 except 3-aininopyridine was used in the place of 2-aminopyridine. MS 513.

Example 14

NH

HN- N 2,3,4,5-Tetrahydro4(1-naphthalenylcarbonyl)7-phenyl1-[3(1H-pyrazol- 3-ylaxnino)propyl]-1H-1,4-benzodiazepine, dihydrochioride.

WO 99/18951 PCTIUS98/19315 34 The title compound was prepared by following the procedure described for the preparation of Example 12 except 3-aminopyrazole was used in the place of 2-aminopyridine. MS 502.

Example

-NS

2,3,4,5-Tetrahydro-4-(1-naphthalenylcarbonyl)-7-phenyl-43-(2pyrimidinylamino)propyll-H-1,4-benzodiazepine, dihydrochioride.

The title compound was prepared by following the procedure described for the preparation of Example 12 except 2-aminopyrimidine was used in the place of 2-aminopyridine. MS 514.

Example 16 Br

NCH

3 (R)-7-Bromo-2,3,4,5-tetrahydro-4-(methyLsulfonyl)-1-(3-pyridinylmethyl)- 2 0 3-(phenyhnethyl)-1H-1,4-benzodiazepine, hydrochloride.

WO 99/18951 PCT/US98/19315 The title compound was prepared from the compound C of Example 4 and nicotinaldehyde by following the procedure for the preparation of Example 6. MS 486.

Example 17 Br

I-

CH3

N

(R)-7-Bromo-2,3,4,5-tetrahydro-4-(methylsulfonyl)-l-(1.oxo-3pyridinylmethyl)-3-(phenylmethyl)-1H-1,4-benzodiazepine, hydrochloride.

To a stirred solution of Example 16 (25 mg, 0.05 mmol) in methylene chloride in the presence of sodium bicarbonate was added mCPBA (50 mg) at o0C. The mixture was stirred for 3 h, and treated with aq. potassium carbonate. The suspension was stirred, filtered, and concentrated. The residue was dissolved in methanol, 1N HC1 in ether was added, solvent was removed to give title compound as a solid (22 mg). MS 502.

WO 99/18951 PCTIUS98/19315 36 Example 18 Br N-SS' CH 3 (R)-7-Bromo-2,34,5-tetrahydro-4(methylsulfonyl)-1.(2-pyridinylmethyl)- 3-(phenyhmethyl)-1H-1,4-benzodiazepine, hydrochloride.

The title compound was prepared from the compound C of Example 4 and 2-pyridylcarboxaldehyde by following the procedure for the preparation of Example 6. MS 486.

Example 19 Br N CH 3 2500 (R)-7-Bromo-2,3,4,5-tetrahydro-4-(nethylsulfonyl)-l-(4-pyridinymethyl)- 3-(phenylmethyl)-lH-1,4-benzodiazepine, hydrochloride.

The title compound was prepared from the compound C of Example 4 and 4-pyridylcarboxaldehyde by following the procedure for the preparation of Example 6. MS 486.

WO 99/18951 PCTIUS98/19315 37 Example Br 000 (R)-7-Bromo-2,3,4,5-tefrahydro.N-hydroxy4(methylsufonyl).1H1,4 benzodiazepine-1-acetamide, hydrochloride A. Ethyl (R)-7-Bromo-2,3,4,5-tetrahydro-4(methysulfonyl)-1H-1,4.

benzodiazepine-1-acetate The title compound was prepared from the compound C of Example 4 and ethyl glyoxalate by following the procedure for the preparation of Example 6. MS 481.

B. (R)-7-Bromo-2,3,4,5-tetrahydro-4-(methyLsufonyl)-1H-1,4benzodiazepine-1-acetic acid.

To a stirred solution of compound A (480 mg, 1.0 mmol) in THF in the presence of aq. methanol, was added LiOH*H 2 0 (210 mg, 5.0 mmol).

The mixture was allowed to stir for 3 h, and made acidic with 1 N HCl solution. The mixture was concentrated in vacuo, and partitioned between ethyl acetate and 1 N HCl solution. The organic layer was separated, dried, and concentrated to give the title compound (400 mg, MS 451.

WO 99/18951 PCTIUS98/1 9315 38 C. 7 -Bromo-2,3,4,5-tetrahydro-N-hydroxy-4-(methylsulfonyl)-lH- 1,4-benzodiazepine-l-acetamide, hydrochloride To a stirred solution of compound B (230 mg, 0.5 mmol) in DMF, .was added hydroxylamine hydrochloride (350 mg, 5.0 mmol), triethylamine (0.07 mL, 0.5 mmol) and 1-(3-dimethylaminopropyl)-3ethylcarbodiimide hydrochloride (100 mg, 0.5 mmol). The mixture was allowed to stir for 18 h, and concentrated in vacuo. The residue was partitioned between ethyl acetate and 1 N HC1 solution. The organic layer was separated, dried, concentrated. The residue was purified by chromatography ethyl acetate, hexanes). The resultant oil was dissolved in methanol, 1 N HC1 was added, the solvent was removed to give title compound. MS 468.

Claims (2)

1. A compound of the formula R"'G and 4 R D N Z-R 8 9 9 9

9. 9 9 9 9 9999 9*99 9* 9* 99 9*99 9 9 a .9 9 R-Rr-Sr-Tt- ii their enantiomers, diastereomers, and pharmaceutically acceptable salts, prodrugs and solvates thereof inhibit farnesyl protein transferase 10 which is an enzyme involved in ras oncogene expression. In formulas I- II and throughout their specification, the above symbols are defined as follows: m, r, s and t are 0 or 1; X is selected from the group consisting of carbon, oxygen, NR 7 or absent. Z is selected from the group consisting of CHR 9 SO 2 CO, C02, 0, NR 1 0 S0 2 NR 1 1 CONR 2 N CN CN0 11 11 11 i-C-N- 1 -N'S02-/ 1 13 R 1 14 R I R 1 -S0 2 R0R 'N NR 2 R 6R7R 18 R' 9 S- I -S- W:ASPECIM~886-98.doc or Z may be absent; Y is selected from the group consisting of CHR 2 3 SO 2 CO, NR 2 4 S0 2 NR 2 5 CONR 2 6 or Y may be absent; R 6 R 7 R 9 R 1 0 R 1 1 R 1 2 R 13 R 1 4 R 1 5 R 1 6 R 1 7 R18, R19, R 2 0 R 2 1 R 2 2 R 2 4 R 2 5 2 6 R 2 8 R 2 9 R 3 0 R 3 1 R 3 2 R 3 3 R 34, R 3 5 R 3 6 R 3 7 R 3 8 R 3 9 R 4 1 R42, R 4 3 R 4 4 R 4 5 and R 4 6 are selected from the group consisting of hydrogen, lower alkyl, substituted alkyl, aryl, or substituted aryl; R 4 R 5 R 4 7 R 4 8 R 4 9 R 5 0 and R 5 1 are selected from the group consisting of hydrogen, halo, nitro, cyano and U-R 2 7 U is selected from the group consisting of sulfur, oxygen, NR 2 8 CO, SO, 502, C0 2 NR 29 C0 2 NR 30 CONR 31 NR 32 SO 2 NR 33 SO 2 NR 34 S0 2 NR 3 5 NR 3 6 CO, CONR 3 7 P0 2 R 3 8 and P0 3 R 3 9 or U is absent; 4 5 R R may join together to form a ring; 15 R 1 R 2 and R3 are selected from the group consisting of hydrogen, alkyl, alkoxycarbonyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aralkyl, cycloalkyl, aryl, substituted aryl, heterocyclo, substituted heterocyclo, cyano, carboxy, carbamyl (e.g. CONH,) or substituted carbamyl further selected from CONH alkyl, 20 CONH aryl, CONH aralkyl or cases where there are two substituents on the nitrogen selected from alkyl, aryl or aralkyl; R 8 and R 2 7 are selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted **)alkynyl, aralkyl, cycloalkyl, aryl, substituted aryl, heterocyclo, 25 substituted heterocyclo; Any two of R 1 R 2 and R 3 may join to form a cycloalkyl group; R, S and T are selected from the group consisting of CR 4 0 R 4 1 NR 4 2 R 4 3 is selected from NR 4 4 R 4 5 OR 4 6 CN; G is selected from the group consisting of -SO 2 NH-, 47 48 0 OH OH SH NC C-N- -NHSO 2 OH N R50 R51 0 R 49 RO N OR 51 II -N-C-N R zOH 1OOH and heterocycles other than imidazole; A, B, C and D are selected from carbon, oxygen, sulfur or nitrogen, or D may be absent. with the provisos that 1. R 2 7 may be hydrogen except when U is SO, SO 2 C0 2 NR 2 5 C0 2 or NR 2 8 S0 2 2. R 8 may be hydrogen except when Z is SO 2 C0 2 or 0 NR 20 R 21 N -NR 22 -S0 2 S- -s- R 1 3. 4. Only one of Y, R, S and T may be nitrogen, Any one of Y, R, S, and T may be nitrogen except when G is 0 0 F1 49 11 11 -N-C-N -NHSO 2 OH OHi R 6 may be hydrogen except when G is NHSO 2 6. G may be imidazole only if Xis oxygen or NR 7 and 7. Three of r, s, and t together may not be zero. 2. The compound of claim 1 wherein AiBCD and X are carbons, namely 2,3,4 3. The compound of claims 1 or 2 wherein m is zero. 4. The compound of claim 1 selected from the groups consisting of 1-(2-Amino-3-mercaptoproopyl)-2,3 ,4,5-tetrahydro-4- (naphthalenylcarbonyl)- 1H- 1,4-benzodiazepine, monohydrochloride 1-(2-Amino-3-mercaptopropyl)-2,3 ,4 ,5-tetrahydro-4- (naphthalenylcarbonyl)-7-phenyl- 1H-1,4-benzodiazepine, mon ohydro chloride 2,3 ,4,5-Tetrahydro-4-( 1-naphthalenylsulfonyl )-N-(3-pyridinyl)- 1H- 1,4- benzodiazepine-1-carboxamide WO) 99/18R951 PCTIUS98Q/1 9315~ 42 (R)-7-Bromo-2,3 ,4,5-tetrahydro-4-(methylsulfonyl)-1- [(2-oxo- 1- pyrrolidinyl)acetyl] -3-(phenylmethyl)- 1H- 1,4-benzodiazepine, trifluoroacetate. (R)-7-Bromo-4- [12-(dimethylamino)ethyl] sulfonyll -2,3 oxo-3-(phenylmethyl)- 1H- 1,4-benzodiaze-pine- 1-ethanesulfonamide. 2,3 ,4,5-Tetrahydro-4-( 1-naphthalenylcarbonyl)- 1-(2-thienylmethyl)- 1H- 1,4-benzodiazepine. 2,3 ,4,5-Tetrahydro-4-( 1-naphthalenylcarbonyl)- 1-(3-pyridinylmethyl)-7- phenyl- 1H- 1,4-benzodiazepine, hydrochloride. 2,3,4, 5-Tetrahydro-4-( 1-naphthalenylcarbonyl)- 1-(3-thienylmethyl)- 1H- 1,4-benzodiazepine 2,3,4,5-Tetrahydro-1- [2-(3-hydroxy-2-oxo- 1(2H)-pyridinyl)ethyl] 1- naphthalenylcarbonyl)- 1H- 1,4-benzodiazepine, dihydrochioride. 2,3,4,5-Tetrahydlro-1- [2-(3-hydroxy-2-oxo- 1(2H)-pyridinyl)ethyll 1- naphthalenylcarbonyl)-7-phenyl- 1H-1 ,4-benzodiazepine, dihydrochioride. 2,3 ,4,5-Tetrahydro-4-( 1-naphthalenylcarbonyl)-1- [(5-oxo-2- pyrrolidinyl)carbonyl] -7-phenyl- 1H- 1,4-benzodiazepine. 2,3 ,4,5-Tetrahydro-4-( 1-naphthalenylcarbonyl)-7-phenyl-1- pyridinylamino)propyll i- 1,4-benzodiazepine, dihydrochioride. 2 ,3,4,5-Tetrahydro-4-( 1-naphthalenylcarbonyl)-7-phenyl-1- pyridinylamino)propyl] H- 1,4-benzodiazepine, dihydrochioride. 2,3 ,4,5-Tetrahydro-4-( 1-naphthalenylcarbonyl)-7-phenyl-1- 1H-pyrazob- 3-ylamino)propyll H- 1,4-benzodiazepine, dihydrochioride. 2,3,4, 5-Tetrahydro-4-(l1-naphthalenylcarbony)-7-phenyl pyrimidinylamino)propyl] H-1 ,4-benzodiazepine, dihydrochioride. -B rom o- 2,3 ,4,5 -tetrahydro-4-(m ethyl sul fonyl)- 14(3-pyridinylme thyl)- 3 -(phenylmethyl)-1H-1,4-benzodiazepine, hydrochloride. 7 -Bromo-2,3,4,5-tetrahydiro.4-(methylsulfonyl)y1(l>oxo-3 pyridinylmethyl)-3-(phenylm ethyl)- 1 H-i ,4-benzodiazepine, hydrochloride. (R)-7-Brorno-2,3 ,4,5-tetrahydro-N-hydroxy-4-(methylsulfonyl).iH-.1,4- benzodiazepine-l1-acetamide, hydrochloride (R)-7-Bromo-2,3 ,4,5-tetrahydro-4-(methylsulfonyl)- 1-(2-pyridinylmethvl)- 3-(phenylmethyl)- 1H- 1,4-benzodiazepine, hydrochloride. 7-B romno- 2,3 ,4,5 -tetrahydro-4 -(mnethylsul fonyl)- 1- (4-pyri dinylm ethyl)- 3-(phenylmethyl)- 1H-1 ,4-benzodiazepine, hydrochloride. A method of inhibiting farnesyl protein transferase which comprises administering to a mammalian subject an effective farnesyl protein transferase inhibiting amount of a compound of any one of claims 1 -4. 6. A method of inhibiting prenyl transferases which comprises administering to a mamm alian subject an effective prenyl transferase inhibiting amount of a compound of any one of claims 1-4. 7. A method of inhibiting tumors which comprises administering to a mammalian subject an effective tumor inhibiting amount of a compound of any one of claims 1-4. ~JC~RA 8. A method of treating diseases associated with signal transduction pathways operating through Ras which comprises 44 administering to a manmmalian subject an amount of a compound of any one of claims 1-4 effective for treating said diseases. 9. A method of treating diseases associated with proteins that are posttranslationally modified by the enzyme farnesyl protein transferase which comprises administering to a mammalian subject an amount of a compound of any one of claims 1-4 effective for treating said diseases. A method of treating disease associated with proteins that are posttranslationally modified by the enzymes geranylgeranyl protein transferase which comprises administering to a mammalian subject an amount of a compound of any one of claims 1-4 effective for treating said diseases. DATED: 7 July 2000 99*9 PHILLIPS ORMONDE FITZPATRICK Patent Attorneys for: BRISTOL-MYERS SQUIBB COMPANY o *o g W:\SPECI\94886-98.doc