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WO1997027752A1 - Inhibiteurs de la farnesyl-proteine transferase - Google Patents

Inhibiteurs de la farnesyl-proteine transferase Download PDF

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Publication number
WO1997027752A1
WO1997027752A1 PCT/US1997/001599 US9701599W WO9727752A1 WO 1997027752 A1 WO1997027752 A1 WO 1997027752A1 US 9701599 W US9701599 W US 9701599W WO 9727752 A1 WO9727752 A1 WO 9727752A1
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Prior art keywords
hydrogen
substituted
alkyl
aryl
unsubstituted
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PCT/US1997/001599
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English (en)
Inventor
Christopher J. Dinsmore
George D. Hartman
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Merck & Co., Inc.
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Publication date
Priority claimed from GBGB9605699.9A external-priority patent/GB9605699D0/en
Application filed by Merck & Co., Inc. filed Critical Merck & Co., Inc.
Priority to EP97904147A priority Critical patent/EP0880320A4/fr
Priority to AU18516/97A priority patent/AU712504B2/en
Publication of WO1997027752A1 publication Critical patent/WO1997027752A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles

Definitions

  • Ras proteins are part of a signalling pathway that links cell surface growth factor receptors to nuclear signals initiating cellular proliferation.
  • Biological and biochemical studies of Ras action indicate that Ras functions like a G-regulatory protein.
  • Ras In the inactive state, Ras is bound to GDP.
  • Ras Upon growth factor receptor activation Ras is induced to exchange GDP for GTP and undergoes a conformational change.
  • the GTP-bound form of Ras propagates the growth stimulatory signal until the signal is
  • Mutated ras genes (Ha-ras, Ki4a-r ⁇ s, Ki4b-ras and N-ras) are found in many human cancers, including colorectal carcinoma, exocrine pancreatic carcinoma, and myeloid leukemias. The protein products of these genes are defective in their GTPase activity and constitutively transmit a growth stimulatory signal.
  • Ras C-terminus contains a sequence motif termed a "CAAX” or "Cys-Aaa 1 -Aaa 2 -Xaa” box (Cys is cysteine, Aaa is an aliphatic amino acid, the Xaa is any amino acid) (Willumsen et al., Nature 310:583-586 (1984)).
  • this motif serves as a signal sequence for the enzymes farnesyl-protein transferase or geranylgeranyl-protein transferase, which catalyze the alkylation of the cysteine residue of the CAAX motif with a C 15 or C 20 isoprenoid, respectively.
  • the Ras protein is one of several proteins that are known to undergo post-translational farnesylation.
  • farnesylated proteins include the Ras-related GTP-binding proteins such as Rho, fungal mating factors, the nuclear lamins, and the gamma subunit of transducin. James, et al., J. Biol. Chem. 269, 14182 (1994) have identified a peroxisome associated protein Pxf which is also farnesylated. James, et al., have also suggested that there are Ras-related GTP-binding proteins such as Rho, fungal mating factors, the nuclear lamins, and the gamma subunit of transducin. James, et al., J. Biol. Chem. 269, 14182 (1994) have identified a peroxisome associated protein Pxf which is also farnesylated. James, et al., have also suggested that there are
  • FPTase farnesyl-protein transferase
  • FPP farnesyl diphosphate
  • Ras protein substrates
  • the peptide derived inhibitors that have been described are generally cysteine containing molecules that are related to the CAAX motif that is the signal for protein prenylation. (Schaber et al, ibid; Reiss et. al, ibid; Reiss et al., PNAS, 88:732-736 (1991)).
  • Such inhibitors may inhibit protein prenylation while serving as alternate substrates for the farnesyl-protein transferase enzyme, or may be purely competitive inhibitors (U.S.
  • Patent 5,141,851 University of Texas; N.E. Kohl et al, Science,
  • transferase inhibitors are inhibitors of proliferation of vascular smooth muscle cells and are therefore useful in the prevention and thereapy of arteriosclerosis and diabetic disturbance of blood vessels (JP H7-112930).
  • the present invention comprises small molecule peptidomimetic urea-containing compounds which inhibit the farnesyl-protein transferase.
  • the instant compounds lack a thiol moiety and thus offer unique advantages in terms of improved pharmacokinetic behavior in animals, prevention of thiol-dependent chemical reactions, such as rapid autoxidation and disulfide formation with endogenous thiols, and reduced systemic toxicity.
  • the compounds of this invention are useful in the inhibition of farnesyl-protein transferase and the farnesylation of the oncogene protein Ras.
  • the inhibitors of farnesyl-protein transferase are illustrated by the formula I: wherein:
  • R 1a , R 1b and R 2 are independently selected from:
  • R 3 and R 4 are independently selected from F, Cl, Br, N(R 8 ) 2 , CF 3 ,
  • R 5a and R 5b are independently selected from:
  • R 6 is independently selected from:
  • R 8 OC(O)-, N 3 , -N(R 8 ) 2 , or R 9 OC(O)NR 8 -, and c) C 1 -C 6 alkyl unsubstituted or substituted by aryl,
  • R 7 is selected from:
  • V is selected from:
  • V is not hydrogen if A 1 is S(O) m and V is not hydrogen if A 1 is a bond, n is 0 and A 2 is S(O) m ; W is a heterocycle;
  • Y is aryl or heteroaryl; m is 0, 1 or 2;
  • n 0, 1, 2, 3 or 4;
  • p 0, 1, 2, 3 or 4;
  • r is 0 to 5, provided that r is 0 when V is hydrogen; and t is 0 or 1; or the pharmaceutically acceptable salts thereof.
  • R 1a and R 2 are independently selected from: hydrogen or C 1 -C 6 alkyl;
  • R 1b is independently selected from:
  • heterocycle cycloalkyl, alkenyl, R 8 O-, or -N(R 8 ) 2 ;
  • R 3 and R 4 are independently selected from F, Cl, Br, N(R 8 ) 2 , CF 3 ,
  • R 5a and R 5b are independently selected from:
  • R 6 is independently selected from:
  • perfluoroalkyl F, Cl, R 8 O-, R 8 C(O)NR 8 -, CN, NO 2 , (R 8 ) 2 N-C(NR 8 )-, R 8 C(O)-, R 8 OC(O)-, -N(R 8 ) 2 , or R 9 OC(O)NR 8 -, and
  • R 7a is hydrogen or methyl
  • R 8 is independendy selected from hydrogen, C 1 -C 6 alkyl, benzyl and aryl
  • R 9 is independendy selected from C 1 -C 6 alkyl and aryl
  • V is selected from:
  • heterocycle selected from pyrrolidinyl, imidazolyl,
  • V is not hydrogen if A 1 is S(O) m and V is not hydrogen if A 1 is a bond, n is 0 and A 2 is S(O) m ; m is 0, 1 or 2;
  • n 0, 1, 2, 3 or 4;
  • p 0, 1, 2, 3 or 4;
  • r is 0 to 5, provided that r is 0 when V is hydrogen; and or the pharmaceutically acceptable salts thereof.
  • R 1 a and R 2 are independently selected from: hydrogen or C 1 -C 6 alkyl
  • R 1b is independendy selected from:
  • heterocycle cycloalkyl, alkenyl, R 8 O-, or -N(R 8 ) 2 ;
  • R 3 and R 4 are independently selected from F, Cl, Br, N(R 8 ) 2 , CF 3 ,
  • R 5a and R 5b are independently selected from:
  • R 6 is independendy selected from:
  • perfluoroalkyl F, Cl, R 8 O-, R 8 C(O)NR 8 -, CN, NO 2 , (R 8 ) 2 N-C(NR 8 )-, R 8 C(O)-, R 8 OC(O)-, -N(R 8 ) 2 , or R 9 OC(O)NR 8 -, and
  • R 7 is selected from: hydrogen and C 1 -C 6 alkyl;
  • R 8 is independendy selected from hydrogen, C 1 -C 6 alkyl, benzyl and aryl;
  • R 9 is independendy selected from C 1 -C 6 alkyl and aryl;
  • V is selected from: a) hydrogen
  • heterocycle selected from pyrrolidinyl, imidazolyl,
  • V is not hydrogen if A 1 is S(O) m and V is not hydrogen if A 1 is a bond, n is 0 and A 2 is S(O) m ;
  • W is a heterocycle selected from pyrrolidinyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or isoquinolinyl; m is 0, 1 or 2;
  • n 0, 1, 2, 3 or 4;
  • p 0, 1, 2, 3 or 4;
  • r is 0 to 5, provided that r is 0 when V is hydrogen; and t is 1; or the pharmaceutically acceptable salts thereof.
  • the inhibitors of farnesyl-protein transferase are illustrated by the formula Ic:
  • R 1b is independently selected from:
  • R 2 are independently selected from: hydrogen or C 1 -C 6 alkyl
  • R 3 and R 4 are independendy selected from F, Cl, Br, N(R 8 ) 2 , CF 3 ,
  • R 5a and R 5b are independently selected from:
  • R 6 is independendy selected from:
  • R 8 perfluoroalkyl, F, Cl, R 8 O-, R 8 C(O)NR 8 -, CN, NO 2 , (R 8 ) 2 N-C(NR 8 )-, R 8 C(O)-, R 8 OC(O)-, -N(R 8 ) 2 , or R 9 OC(O)NR 8 -, and c) C 1 -C 6 alkyl substituted by C 1 -C 6 perfluoroalkyl, R 8 O-, R 8 C(O)NR 8 -, (R 8 ) 2 N-C(NR 8 )-, R 8 C(O)-, R 8 OC(O)-, -N(R 8 ) 2 , or R 9 OC(O)NR 8 -;
  • R 8 is independendy selected from hydrogen, C 1 -C 6 alkyl, benzyl and aryl;
  • p 0, 1, 2, 3 or 4; or the pharmaceutically acceptable salts thereof.
  • the inhibitors of farnesyl-protein transferase are illustrated by the formula Id:
  • R 1b is independendy selected from:
  • R 2 are independently selected from: hydrogen or C 1 -C 6 alkyl
  • R 3 and R 4 are independently selected from F, Cl, Br, N(R 8 ) 2 , CF 3 ,
  • R 5a and R 5b are independently selected from:
  • p 0, 1, 2, 3 or 4; or the pharmaceutically acceptable salts thereof.
  • the compounds of the present invention may have asymmetric centers and occur as racemates, racemic mixtures, and as individual diastereomers, with all possible isomers, including optical isomers, being included in the present invention.
  • any variable e.g. aryl, heterocycle, R 1a , R 2 etc.
  • its definition on each occurence is independent at every other occurence.
  • combinations of substituents/or variables are permissible only if such combinations result in stable compounds.
  • alkyl is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms; “alkoxy” represents an alkyl group of indicated number of carbon atoms attached through an oxygen bridge.
  • Halogen or “halo” as used herein means fluoro, chloro, bromo and iodo.
  • aryl is intended to mean any stable monocyclic or bicyclic carbon ring of up to 7 members in each ring, wherein at least one ring is aromatic.
  • aryl elements include phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthryl, anthryl or acenaphthyl.
  • heterocycle or heterocyclic represents a stable 5- to 7-membered monocyclic or stable 8- to 11- membered bicyclic heterocyclic ring which is either saturated or unsaturated, and which consists of carbon atoms and from one to four heteroatoms selected from the group consisting of N, O, and S, and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring.
  • the heterocyclic ring may be attached at any heteroatom or carbon atom which results in the creation of a stable structure.
  • heterocyclic elements include, but are not limited to, azepinyl, benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl,
  • heteroaryl is intended to mean any stable monocyclic or bicyclic carbon ring of up to 7 members in each ring, wherein at least one ring is aromatic and wherein from one to four carbon atoms are replaced by heteroatoms selected from the group consisting of N, O, and S.
  • heterocyclic elements include, but are not limited to, benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl,
  • Lines drawn into the ring systems from substituents indicate that the indicated bond may be attached to any of the substitutable ring carbon atoms.
  • R 1a , R 1b and R 2 are independendy selected from: hydrogen, -N(R 8 ) 2 , R 8 C(O)NR 8 - or C 1 -C 6 alkyl unsubstituted or substituted by -N(R 8 ) 2 , R 8 O- or R 8 C(O)NR 8 -.
  • R 3 and R 4 are independently selected from: hydrogen, perfluoroalkyl, F, Cl, Br, R 8 O-, R 9 S(O) m -, CN, NO 2 , R 8 2 N-C(NR 8 )-, R 8 C(O)-, R 8 OC(O)-, N 3 , -N(R 8 ) 2 , or R 9 OC(O)NR 8 - and C 1 -C 6 alkyl.
  • R 5a and R 5b are independently selected from hydrogen or C 1 -C 6 alkyl substituted with hydrogen, R 9 S(O) m -, CF 3 - or an unsubstituted or substituted aryl group.
  • R 6 is selected from: hydrogen, perfluoroalkyl,
  • R 7 is hydrogen or methyl. Most preferably,
  • R 7 is hydrogen
  • R 8 is selected from H, C 1 -C 6 alkyl and benzyl.
  • a 1 and A 2 are independently selected from: a bond, -C(O)NR 8 -, -NR 8 C(O)-, O, -N(R 8 )-, -S(O) 2 N(R 8 )- and-
  • V is selected from hydrogen, heterocycle and aryl. Most preferably, V is phenyl.
  • Y is selected from phenyl, pyridyl, furyl and thienyl. Most preferably, Y is phenyl.
  • n, p and r are independendy 0, 1, or 2.
  • t is 1.
  • the pharmaceutically acceptable salts of the compounds of this invention include the conventional non-toxic salts of the compounds of this invention as formed, e.g., from non-toxic inorganic or organic acids.
  • such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like: and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic and the like.
  • -N(R 8 ) 2 represents -NHH, -NHCH 3 , -NHC 2 H 5 , etc. It is understood that substituents and substitution patterns on the compounds of the instant invention can be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be readily synthesized by techniques known in the art, as well as those methods set forth below, from readily available starting materials.
  • the pharmaceutically acceptable salts of the compounds of this invention can be synthesized from the compounds of this invention which contain a basic moiety by conventional chemical methods.
  • the salts are prepared either by ion exchange
  • Schemes 1-3 illustrates the synthesis of one of the preferred embodiments of the instant invention, wherein the variable W is present as a imidazolyl moiety that is substituted with a suitably substituted benzyl group.
  • Substituted protected imidazole alkanols II can be prepared by methods known in the art, such as those described by F. Schneider, Z. Physiol. Chem., 3:206-210 (1961) and C.P. Stewart, Biochem. Journal, 17:130-133(1923).
  • reagent may be ring alkylated to provide the intermediate amine VIII.
  • Amines such as those illustrated in Schemes 1 and 2 may be reacted with a suitably substituted isocyanate IX to provide the instant compound X.
  • Compound X can be selectively N-alkylated under standard conditions, such as those illustrated, to provide the instant compound XI.
  • the isocyanate IX can be reacted with a variety of other amines, such as XII, as shown in Scheme 8.
  • the product XIII can be deprotected to give the instant compound XIV.
  • the compound XIV is isolated in the salt form, for example, as a trifluoroacetate,
  • Compound XIV can further be selectively protected to obtain XV which can subsequentiy be reductively alkylated with a second aldehyde, such as XVI, to obtain XVII. Removal of the protecting group, and conversion to cyclized products such as the dihydroimidazole XIX can be accomplished by literature procedures.
  • a bis-protected aldehyde XX may be reacted with a suitable Grignard reagent to provide the secondary alcohol XXI. Subsequent protection and reductive deprotection provides the primary alcohol XXII. This alcohol can then be converted to the corresponding amine by the techniques illustrated in Schemes 1-2 above. Amine XXIII may then be reacted with isocyanate DC to provide the carbamate XXIV.
  • a fully deprotected amino alcohol XXVII can be reductively alkylated (under conditions described previously) with a variety of aldehydes to obtain secondary amines, such as XXVIII (Scheme 10), or tertiary amines.
  • Boc protected amino alcohol XXIX can also be utilized to synthesize 2-aziridinylmethylureas such as XXX (Scheme 11).
  • Treating XXIX with 1,1'-sulfonyldiimidazole and sodium hydride in a solvent such as dimethylformamide can lead to the formation of aziridine XXX.
  • the aziridine can be reacted with a nucleophile, such as a thiol, in the presence of base to yield the ring-opened product XXXI.
  • isocyanate IX can be reacted with aldehydes derived from amino acids such as O-alkylated tyrosines, according to standard procedures, to obtain compounds such as XXXVII, as shown in Scheme 12.
  • R' is an aryl group
  • XXXVII can first be
  • XXXVIII hydrogenated to unmask the phenol, and the amine group deprotected with acid to produce XXXVIII.
  • the amine protecting group in XXXVII can be removed, and O-alkylated phenolic amines such as IXL produced.
  • Schemes 13 illustrates an alternate preparation of the instant compounds. As shown in Scheme 13, the isocyanate XL is formed first and is then treated with the suitably substituted aniline such as XLI to provide the instant compound X.
  • the instant compounds are useful as pharmaceutical agents for mammals, especially for humans. These compounds may be administered to patients for use in the treatment of cancer. Examples of the type of cancer which may be treated with the compounds of this invention include, but are not limited to, colorectal carcinoma, exocrine pancreatic carcinoma, myeloid leukemias and neurological tumors.
  • Such tumors may arise by mutations in the ras genes themselves, mutations in the proteins that can regulate Ras formation (i.e.,
  • NF-1 neurofibromen
  • neu neu
  • scr abl
  • lck lck
  • fyn neurofibromen
  • the compounds of the instant invention inhibit farnesyl-protein transferase and the farnesylation of the oncogene protein Ras.
  • the instant compounds may also inhibit tumor angiogenisis, thereby affecting the growth of tumors (J. Rak et al. Cancer Research, 55:4575-4580 (1995)).
  • tumor angiogenisis Such anti-angiogenisis properties of the instant
  • the compounds of this invention are also useful for inhibiting other proliferative diseases, both benign and malignant, wherein Ras proteins are aberrantiy activated as a result of oncogenic mutation in other genes (i.e., the Ras gene itself is not activated by mutation to an oncogenic form) with said inhibition being accomplished by the administration of an effective amount of the compounds of the invention to a mammal in need of such treatment.
  • Ras proteins are aberrantiy activated as a result of oncogenic mutation in other genes (i.e., the Ras gene itself is not activated by mutation to an oncogenic form) with said inhibition being accomplished by the administration of an effective amount of the compounds of the invention to a mammal in need of such treatment.
  • a component of NF-1 is a benign proliferative disorder.
  • the instant compounds may also be useful in the treatment of certain viral infections, in particular in the treatment of hepatitis delta and related viruses (J.S. Glenn et al. Science, 256:1331-1333 (1992).
  • the compounds of the instant invention are also useful in the prevention of restenosis after percutaneous transluminal coronary angioplasty by inhibiting neointimal formation (C. Indolfi et al. Nature Medicine, 1:541-545(1995).
  • the instant compounds may also be useful in the treatment and prevention of polycystic kidney disease (D.L. Schaffner et al. American Journal of Pathology, 142:1051-1060 (1993) and B. Cowley, Jr. et al.FASEB Journal, 2:A3160 (1988)).
  • the compounds of this invention may be administered to mammals, preferably humans, either alone or, preferably, in
  • the compounds can be administered orally or parenterally, including the intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical routes of administration.
  • the selected compound may be administered, for example, in the form of tablets or capsules, or as an aqueous solution or suspension.
  • carriers which are commonly used include lactose and com starch, and lubricating agents, such as magnesium stearate, are commonly added.
  • useful diluents include lactose and dried com starch.
  • the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring agents may be added.
  • sterile solutions of the active ingredient are usually prepared, and the pH of the solutions should be suitably adjusted and buffered.
  • the total concentration of solutes should be controlled in order to render the preparation isotonic.
  • the present invention also encompasses a pharmaceutical composition useful in the treatment of cancer, comprising the
  • compositions of this invention include aqueous solutions comprising compounds of this invention and pharmacologically acceptable carriers, e.g., saline, at a pH level, e.g., 7.4.
  • pharmacologically acceptable carriers e.g., saline
  • the solutions may be introduced into a patient's intramuscular blood-stream by local bolus injection.
  • the daily dosage will normally be determined by the prescribing physician with the dosage generally varying according to the age, weight, and response of the individual patient, as well as the severity of the patient's symptoms.
  • a suitable amount of compound is administered to a mammal undergoing treatment for cancer.
  • Administration occurs in an amount between about 0.1 mg/kg of body weight to about 60 mg/kg of body weight per day, preferably of between 0.5 mg/kg of body weight to about 40 mg/kg of body weight per day.
  • the compounds of the instant invention are also useful as a component in an assay to rapidly determine the presence and quantity of farnesyl-protein transferase (FPTase) in a composition.
  • FPTase farnesyl-protein transferase
  • composition to be tested may be divided and the two
  • mixtures which comprise a known substrate of FPTase (for example a tetrapeptide having a cysteine at the amine terminus) and farnesyl pyrophosphate and, in one of the mixtures, a compound of the instant invention.
  • FPTase for example a tetrapeptide having a cysteine at the amine terminus
  • farnesyl pyrophosphate for example a tetrapeptide having a cysteine at the amine terminus
  • the chemical content of the assay mixtures may be determined by well known
  • inhibitors of FPTase absence or quantitative reduction of the amount of substrate in the assay mixture without the compound of the instant invention relative to the presence of the unchanged substrate in the assay containing the instant compound is indicative of the presence of FPTase in the composition to be tested.
  • potent inhibitor compounds of the instant invention may be used in an active site titration assay to determine the quantity of enzyme in the sample.
  • a series of samples composed of aliquots of a tissue extract containing an unknown amount of farnesyl-protein transferase, an excess amount of a known substrate of FPTase (for example a tetrapeptide having a cysteine at the amine terminus) and farnesyl pyrophosphate are incubated for an appropriate period of time in the presence of varying concentrations of a compound of the instant invention.
  • concentration of a sufficiendy potent inhibitor i.e., one that has a Ki substantially smaller than the
  • concentration of enzyme in the assay vessel required to inhibit the enzymatic activity of the sample by 50% is approximately equal to half of the concentration of the enzyme in that particular sample.
  • Step 4 Preparation of 1-(4-cyanobenzyl)-5-(hydroxymethyl)- imidazole (5)
  • a solution of the acetate 4 (50.4 g) in 1.5 L of 3:1 THF/water at 0 °C was added lithium hydroxide monohydrate (18.9 g).
  • the reaction was concentrated in vacuo, diluted with EtOAc (3 L), and washed with water, sat. aq. NaHCO 3 and brine.
  • the solution was then dried (Na 2 SO 4 ), filtered, and concentrated in vacuo to provide the crude product (26.2 g) as a pale yellow fluffy solid which was sufficiendy pure for use in the next step without further
  • Step 5 Preparation of 1-(4-cyanobenzyl)-5-imidazolecarboxaldehyde (6)
  • Step 6 Preparation of 1-(4-cyanobenzyl)-5-(n-pentylaminomethyl)imidazole (7)
  • Step 7 Preparation of N-(3-chlorophenyl)-N'-[1-(4-cyanobenzyl)- 5-imidazolylmethyl]-N'-(n-pentyl)urea hydrochloride (1)
  • NaH 37 mg, 60% dispersion in mineral oil
  • 3-Chlorophenylisocyanate 0.084 mL was added dropwise, and the cooling bath was removed.
  • the reaction was poured into EtOAc/hexane (2:1) and water, washed with sat. aq. NaHCO 3 and brine, dried (Na 2 SO 4 ), filtered, and concentrated in vacuo to provide the crude urea 1 as a yellow foam.
  • Half of this material was purified by silica gel chromatography (3-5%
  • urea 1 (remaining half of crude product prepared above) in 1 mL of dry DMF at 0 °C was added NaH (14 mg, 60% dispersion in mineral oil). After 15 minutes, iodomethane (0.029 mL) was added dropwise. The reaction was stirred at 0 °C for four hours, then poured into EtOAc/hexane (2:1) and water, washed with sat. aq. NaHCO 3 and brine, dried (Na 2 SO 4 ), filtered, and concentrated in vacuo to provide a purple oil.
  • BovineFPTase was assayed in a volume of 100 ⁇ l containing 100 mM N-(2-hydroxy ethyl) piperazine-N'-(2-ethane sulfonic acid) (HEPES), pH 7.4, 5 mM MgCl 2 , 5 mM dithiothreitol (DTT), 100 mM [3H]-farnesyl diphosphate ([ 3 H]-FPP; 740 CBq/mmol, New England Nuclear), 650 nM Ras-CVLS and 10 ⁇ g/ml FPTase at 31°C for 60 min. Reactions were initiated with FPTase and stopped with 1 ml of 1.0 M HCL in ethanol.
  • Precipitates were collected onto filter-mats using a TomTec Mach II cell harvestor, washed with 100% ethanol, dried and counted in an LKB ⁇ -plate counter.
  • the assay was linear with respect to both substrates, FPTase levels and time; less than 10% of the [ 3 H]-FPP was utilized during the reaction period.
  • Purified compounds were dissolved in 100% dimethyl sulfoxide (DMSO) and were diluted 20-fold into the assay. Percentage inhibition is measured by the amount of
  • Human FPTase was prepared as described by Omer et al., Biochemistry 32:5167-5176 (1993). Human FPTase activity was assayed as described above with the exception that 0.1% (w/v)
  • Examples 1 and 2 were tested for inhibitory activity against human FPTase by the assay described above and were found to have IC50 of ⁇ 10 ⁇ M.
  • the cell line used in this assay is a v-ras line derived from either Ratl or N1H3T3 cells, which expressed viral Ha-ras p21.
  • the assay is performed essentially as described in DeClue, J.E. et al., Cancer Research 51:712-717, (1991). Cells in 10 cm dishes at 50-75%
  • the cells are labelled in 3 ml methionine-free DMEM supplemeted with 10% regular DMEM, 2% fetal bovine serum and 400 mCi[ 35 S]methionine (1000 Ci/mmol).
  • the cells are lysed in 1 ml lysis buffer (1% NP40/20 mM HEPES, pH 7.5/5 mM MgCl 2 /1mM DTT/10 mg/ml aprotinen/2 mg/ml leupeptin/2 mg/ml antipain/0.5 mM PMSF) and the ly sates cleared by centrifugation at 100,000 ⁇ g for 45 min. Aliquots of ly sates containing equal numbers of acid-precipitable counts are bought to 1 ml with IP buffer (lysis buffer lacking DTT) and immunoprecipitated with the ras-specific monoclonal antibody Y13-259 (Furth, M.E. et al., J. Virol.
  • Rat 1 cells transformed with either v-ras, v-raf, or v-mos are seeded at a density of 1 ⁇ 10 4 cells per plate (35 mm in diameter) in a 0.3% top agarose layer in medium A (Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum) over a bottom agarose layer (0.6%). Both layers contain 0.1% methanol or an appropriate concentration of the instant compound (dissolved in methanol at 1000 times the final concentration used in the assay).
  • the cells are fed twice weekly with 0.5 ml of medium A containing 0.1% methanol or the concentration of the instant compound.
  • Photomicrographs are taken 16 days after the cultures are seeded and comparisons are made.

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Abstract

Composés inhibants la farnésyl-protéine transférase (FTase) et la farnésylation du Ras protéique oncogène. L'invention porte en outre sur des compositions chimiothérapeutiques renfermant les composés de cette invention et sur des méthodes d'inhibitions de la farnésyl-protéine transférase et de la farnésylation du Ras protéique oncogène.
PCT/US1997/001599 1996-01-30 1997-01-27 Inhibiteurs de la farnesyl-proteine transferase WO1997027752A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP97904147A EP0880320A4 (fr) 1996-01-30 1997-01-27 Inhibiteurs de la farnesyl-proteine transferase
AU18516/97A AU712504B2 (en) 1996-01-30 1997-01-27 Inhibitors of farnesyl-protein transferase

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US1086096P 1996-01-30 1996-01-30
US60/010,860 1996-01-30
GBGB9605699.9A GB9605699D0 (en) 1996-03-19 1996-03-19 Inhibitors of farnesyl-protein transferase
GB9605699.9 1996-03-19

Publications (1)

Publication Number Publication Date
WO1997027752A1 true WO1997027752A1 (fr) 1997-08-07

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1997/001599 WO1997027752A1 (fr) 1996-01-30 1997-01-27 Inhibiteurs de la farnesyl-proteine transferase

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Country Link
EP (1) EP0880320A4 (fr)
AU (1) AU712504B2 (fr)
CA (1) CA2243507A1 (fr)
WO (1) WO1997027752A1 (fr)

Cited By (20)

* Cited by examiner, † Cited by third party
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US5929250A (en) * 1997-01-23 1999-07-27 Smithkline Beecham Corporation IL-8 receptor antagonists
FR2780892A1 (fr) * 1998-07-08 2000-01-14 Sod Conseils Rech Applic Utilisation d'inhibiteurs de prenyltransferases pour preparer un medicament destine a traiter les pathologies qui resultent de la fixation membranaire de la proteine g heterotrimerique
US6331541B1 (en) 1998-12-18 2001-12-18 Soo S. Ko N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6358985B1 (en) 1998-07-02 2002-03-19 Merck & Co., Inc. Inhibitors of prenyl-protein transferase
US6380228B1 (en) 2000-04-10 2002-04-30 Merck & Co., Inc. Inhibitors of prenyl-protein transferase
US6410534B1 (en) 1998-07-02 2002-06-25 Merck & Co., Inc. Inhibitors of prenyl-protein transferase
US6441001B1 (en) 1998-12-18 2002-08-27 Bristol-Myers Squibb Pharma Company 2-substituted-4-nitrogen heterocycles as modulators of chemokine receptor activity
US6444686B1 (en) 1998-12-18 2002-09-03 Brsitol-Myers Squibb Pharma Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6486180B1 (en) 1998-12-18 2002-11-26 Bristol-Myers Squibb Pharma Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6492400B1 (en) 1998-12-18 2002-12-10 Bristol-Myers Squibb Pharma Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6525069B1 (en) 1998-12-18 2003-02-25 Bristol-Myers Squibb Pharma Co. N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6627629B2 (en) 2000-06-30 2003-09-30 Bristol-Myers Squibb Pharma N-ureidoheterocycloalkyl-piperidines as modulators of chemokine receptor activity
JP2003535844A (ja) * 2000-06-06 2003-12-02 アベンティス・ファーマ・ドイチユラント・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツング 第ＶＩＩａ因子阻害剤である（チオ）尿素誘導体、その製造方法およびその使用
US6712992B2 (en) 2000-05-26 2004-03-30 Basf Aktiengesellschaft Liquid-crystalline composition of matter
WO2004039765A1 (fr) * 2002-10-31 2004-05-13 Amersham Biosciences Ab Utilisation de variantes d'urees en tant que ligands a affinite
US6897234B2 (en) 1999-12-17 2005-05-24 Bristol-Myers Squibb Pharma Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6919356B2 (en) 2002-09-26 2005-07-19 Bristol Myers Squibb Company N-substituted heterocyclic amines as modulators of chemokine receptor activity
US6974869B2 (en) 2001-09-18 2005-12-13 Bristol-Myers Squibb Pharma Company Piperizinones as modulators of chemokine receptor activity
US6992091B2 (en) 2002-09-12 2006-01-31 Bristol-Myers Squibb Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
EP2095819A1 (fr) 2008-02-28 2009-09-02 Maastricht University Dérivés de N-benzyle imidazole et leur utilisation comme inhibiteurs de l'aldosterone synthase

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WO1995010516A1 (fr) * 1993-10-15 1995-04-20 Schering Corporation Composes tricycliques a base d'amides et d'uree utiles pour inhiber la fonction de la proteine g et au traitement de maladies proliferatives
EP0675112A1 (fr) * 1994-03-31 1995-10-04 Bristol-Myers Squibb Company Inhibiteurs de farnésyl-protéine-transférase contenant l'imidazole

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See also references of EP0880320A4 *

Cited By (32)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6015908A (en) * 1997-01-23 2000-01-18 Smithkline Beecham Corporation IL-8 receptor antagonists
US6043374A (en) * 1997-01-23 2000-03-28 Smithkline Beecham Corporation Benzisothiazolidine Compounds
US5929250A (en) * 1997-01-23 1999-07-27 Smithkline Beecham Corporation IL-8 receptor antagonists
US6410534B1 (en) 1998-07-02 2002-06-25 Merck & Co., Inc. Inhibitors of prenyl-protein transferase
US6358985B1 (en) 1998-07-02 2002-03-19 Merck & Co., Inc. Inhibitors of prenyl-protein transferase
FR2780892A1 (fr) * 1998-07-08 2000-01-14 Sod Conseils Rech Applic Utilisation d'inhibiteurs de prenyltransferases pour preparer un medicament destine a traiter les pathologies qui resultent de la fixation membranaire de la proteine g heterotrimerique
WO2000002558A1 (fr) * 1998-07-08 2000-01-20 Societe De Conseils De Recherches Et D'applications Scientifiques (S.C.R.A.S.) Utilisation d'inhibiteurs de prenyltransferases pour preparer un medicament destine a traiter les pathologies qui resultent de la fixation membranaire de la proteine g heterotrimerique
US6444686B1 (en) 1998-12-18 2002-09-03 Brsitol-Myers Squibb Pharma Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6525069B1 (en) 1998-12-18 2003-02-25 Bristol-Myers Squibb Pharma Co. N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6441001B1 (en) 1998-12-18 2002-08-27 Bristol-Myers Squibb Pharma Company 2-substituted-4-nitrogen heterocycles as modulators of chemokine receptor activity
US6875776B2 (en) 1998-12-18 2005-04-05 Bristol-Myers Squibb Pharma Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6486180B1 (en) 1998-12-18 2002-11-26 Bristol-Myers Squibb Pharma Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6492400B1 (en) 1998-12-18 2002-12-10 Bristol-Myers Squibb Pharma Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6521592B2 (en) 1998-12-18 2003-02-18 Bristol-Myers Squibb Pharma Co. N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6780857B2 (en) 1998-12-18 2004-08-24 Bristol-Myers Squibb Pharma Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6919368B2 (en) 1998-12-18 2005-07-19 Bristol-Myers Squibb Pharma Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6331541B1 (en) 1998-12-18 2001-12-18 Soo S. Ko N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6706735B2 (en) 1998-12-18 2004-03-16 Bristol-Myers Squibb Pharma Company 2-substituted-4-nitrogen heterocycles as modulators of chemokine receptor activity
US6897234B2 (en) 1999-12-17 2005-05-24 Bristol-Myers Squibb Pharma Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6380228B1 (en) 2000-04-10 2002-04-30 Merck & Co., Inc. Inhibitors of prenyl-protein transferase
US6712992B2 (en) 2000-05-26 2004-03-30 Basf Aktiengesellschaft Liquid-crystalline composition of matter
JP2003535844A (ja) * 2000-06-06 2003-12-02 アベンティス・ファーマ・ドイチユラント・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツング 第ＶＩＩａ因子阻害剤である（チオ）尿素誘導体、その製造方法およびその使用
JP4809570B2 (ja) * 2000-06-06 2011-11-09 サノフィ−アベンティス・ドイチュラント・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツング 第ＶＩＩａ因子阻害剤である（チオ）尿素誘導体、その製造方法およびその使用
US6627629B2 (en) 2000-06-30 2003-09-30 Bristol-Myers Squibb Pharma N-ureidoheterocycloalkyl-piperidines as modulators of chemokine receptor activity
US6949546B2 (en) 2000-06-30 2005-09-27 Bristol-Myers Squibb Pharma Company N-ureidoheterocycloalkyl-piperidines as modulators of chemokine receptor activity
US6974869B2 (en) 2001-09-18 2005-12-13 Bristol-Myers Squibb Pharma Company Piperizinones as modulators of chemokine receptor activity
US7514430B2 (en) 2001-09-18 2009-04-07 Bristol-Myers Squibb Pharma Company Piperizinones as modulators of chemokine receptor activity
US6992091B2 (en) 2002-09-12 2006-01-31 Bristol-Myers Squibb Company N-ureidoalkyl-piperidines as modulators of chemokine receptor activity
US6919356B2 (en) 2002-09-26 2005-07-19 Bristol Myers Squibb Company N-substituted heterocyclic amines as modulators of chemokine receptor activity
WO2004039765A1 (fr) * 2002-10-31 2004-05-13 Amersham Biosciences Ab Utilisation de variantes d'urees en tant que ligands a affinite
US7645903B2 (en) 2002-10-31 2010-01-12 Ge Healthcare Bio-Sciences Ab Use of urea variants as affinity ligands
EP2095819A1 (fr) 2008-02-28 2009-09-02 Maastricht University Dérivés de N-benzyle imidazole et leur utilisation comme inhibiteurs de l'aldosterone synthase

Also Published As

Publication number Publication date
AU712504B2 (en) 1999-11-11
EP0880320A4 (fr) 1999-06-16
AU1851697A (en) 1997-08-22
CA2243507A1 (fr) 1997-08-07
EP0880320A1 (fr) 1998-12-02

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