WO1998000405A1 - Pharmaceutical preparation - Google Patents

Pharmaceutical preparation Download PDF

Info

Publication number
WO1998000405A1
WO1998000405A1 PCT/US1997/011131 US9711131W WO9800405A1 WO 1998000405 A1 WO1998000405 A1 WO 1998000405A1 US 9711131 W US9711131 W US 9711131W WO 9800405 A1 WO9800405 A1 WO 9800405A1
Authority
WO
WIPO (PCT)
Prior art keywords
phenyl
unsubstituted
substituted
alkoxy
alkyl
Prior art date
Application number
PCT/US1997/011131
Other languages
French (fr)
Inventor
Joseph J. Lynch, Jr.
Joseph J. Salata
Original Assignee
Merck & Co., Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GBGB9617894.2A external-priority patent/GB9617894D0/en
Application filed by Merck & Co., Inc. filed Critical Merck & Co., Inc.
Priority to JP10504289A priority Critical patent/JP2000510155A/en
Priority to EP97931437A priority patent/EP0907644A4/en
Priority to CA002257948A priority patent/CA2257948A1/en
Priority to AU35066/97A priority patent/AU722110B2/en
Publication of WO1998000405A1 publication Critical patent/WO1998000405A1/en

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/06—Antiarrhythmics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
    • C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
    • C07D243/10—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems
    • C07D243/12—1,5-Benzodiazepines; Hydrogenated 1,5-benzodiazepines

Definitions

  • the present invention relates to the co-administration, either simultaneously, separately or sequentially of a selective iKs antagonist and a beta-adrenergic receptor blocking agent for use in preventing, treating and terminating cardiac arrhythmias, such as atrial, supraventricular and ventricular ectopy, tachycardia, flutter or fibrillation, including atrial, supraventricular and ventricular arrhythmias resulting from myocardial ischemic injury in a patient in need thereof.
  • This invention also relates to a pharmaceutical formulation which comprises a selective IKs antagonist and a beta- adrenergic receptor blocking agent along with a pharmaceutically acceptable carrier.
  • arrhythmias often occur as complications to cardiac diseases such as myocardial infarction and heart failure. In a serious case, arrhythmias give rise to ventricular fibrillation and can cause sudden death.
  • antiarrhythmic agents are now available on the market, agents exhibiting both satisfactory effects and high safety profiles, are not yet available for patients.
  • antiarrhythmic agents of Class I according to the classification of Vaughan-Williams, which cause a selective inhibition of the maximum velocity of the upstroke of the action potential (Vmax) are inadequate for preventing ventricular fibrillation.
  • Vmax maximum velocity of the upstroke of the action potential
  • they have problems regarding safety, namely, they cause a depression of the myocardial contractility and have a tendency to induce arrhythmias due to an inhibition of the impulse conduction.
  • Beta-adrenergic receptor blocking agent which belong to Class II are of limited value since their effects are either limited to a certain type of arrhythmia or are contraindicated because of their cardiac depressant properties in certain patients with cardiovascular disease. Their safety, however, is higher than that of the antiarrhythmic agents of Class I.
  • Antiarrhythmic agents of Class III are drugs which cause a selective prolongation of the duration of the action potential without a significant depression of the Vmax.
  • drugs in this class were limited to sotalol and amiodarone, both of which have been shown to possess Class IH properties.
  • Sotalol also possesses Class II effects which may cause cardiac depression and be contraindicated in certain susceptible patients. Amiodarone is severely limited by side effects.
  • Drugs of this class are expected to be effective in preventing ventricular fibrillations.
  • Pure Class UI agents by definition, are not considered to cause myocardial depression or an induction of arrhythmias due to the inhibition of the action potential conduction as seen with Class I antiarrhythmic agents.
  • IKs antagonists In the treatment of arrhythmia, IKs antagonists have demonstrated effectiveness when delivered orally in amounts ranging from about 0.01 to about 1 mg per kg of body weight per day, in a single dose or in 2 to 4 divided doses.
  • the activity of the compounds described herein as antiarrhythmic agents is measured by their ability to block the IKs and iKr currents as determined by the following test protocol.
  • Outward potassium currents are measured in single guinea pig ventricular myocytes using a whole-cell voltage clamp technique described in detail elsewhere (Sanguinetti and Jurkiewicz, 1990, Two conn ents of cardiac delayed rectifier K + current: differential sensitivity to block by Class LU antiarrhythmic agents. J. Gen Physiol. 96: 195-215).
  • Myocytes are isolated by enzymatic (collagenase and protease) digestion of Langandorf perfused hearts. Single cells are then voltage clamped using 1 mm square-bore pipettes filled with 0.5 M Kgluconate, 25 mM KC1, 5 mM K(2)ATP. Cells are bathed in a solution containing, in mN: 132 NaCl, 4KC1, 1.2 MgCl2, 10 HEPES, 10 glucose: pH 7.2, temp. 35°C.
  • Test depolarizations are applied as voltage ramps from -85 to -50 mV, and as steps to -10 mV (0.5 s) and +50 mV (1.0 s).
  • I ⁇ i is measured as peak outward current during the voltage ramp.
  • iKr is measured as tail currents upon repolarization from -10 mV to -50 mV.
  • IKs is measured as time-dependent current during the pulse to +50 mV. Currents are measured during control, then after exposure to drug at two different concentrations.
  • the compounds described herein as selective IKs channel antagonists have an IC50 of less than 100 nM as IKs antagonists.
  • the compounds of this invention are at least 10 times more potent in the blockade of IKs than of blockade of IKr-
  • Beta-adrenergic receptor blocking agents are a class of pharmaceutically active compounds which decrease the positive chronotropic, positive inotropic, bronchodilator and vasodilator responses caused by beta-adrenergic receptor agonists. The magnitude of this decreased response is proportional to the existing sympathetic tone and the concentration of beta-blocker at the receptor sites. Beta-adrenergic receptor blockage is said to reduce cardiac output in both healthy subjects and patients with heart disease. While the mechanism of antihypertension effects of beta-adrenergic receptor blocking agents has not been established, possible mechanisms of action include reduction in cardiac output, reduction in plasma renin activity, and central nervous system sympatholytic action.
  • beta-adrenergic receptor antagonists have been shown effective in reducing the incidence of mortality and sudden death in postinfarction patients (Yusaf et al., Prog Cardiovasc Dis 17: 335-371, 1985; Lau et al., N Eng J Med 327: 248-254, 1992).
  • a method for use in preventing, treating and terminating cardiac arrhythmias such as atrial, supraventricular and ventricular ectopy, tachycardia, flutter or fibrillation, including atrial, supraventricular and ventricular arrhythmias resulting from myocardial ischemic injury in a patient in need thereof which comprises the co- administration, either simultaneously, separately or sequentially of a selective IKs antagonist and a beta-adrenergic receptor blocking agent.
  • This invention also relates to a pharmaceutical formulation which comprises a selective IKs antagonist and a beta-adrenergic receptor blocking agent along with a pharmaceutically acceptable carrier.
  • a method for use in preventing, treating and terminating cardiac arrhythmias such as atrial, supraventricular and ventricular ectopy, tachycardia, flutter or fibrillation, including atrial, supraventricular and ventricular arrhythmias resulting from myocardial ischemic injury in a patient in need thereof which comprises the co- administration, either simultaneously, separately or sequentially of a selective IKs antagonist and a beta-adrenergic receptor blocking agent.
  • This invention also relates to a pharmaceutical formulation which comprises a selective IKs antagonist and a beta-adrenergic receptor blocking agent along with a pharmaceutically acceptable carrier.
  • a “selective IKs antagonist” is meant those compounds which when studied in the test disclosed above have an IC50 of less than 100 nM as IKs blockers.
  • the compounds of this invention are at least 10 times more potent in the blockade of IKs than of blockade of IKr-
  • Beta-adrenergic receptor blocking agents are compounds which decrease the positive chronotropic, positive inotropic, bronch- odilator and vasodilator responses caused by beta-adrenergic receptor agonists. The magnitude of this decreased response is proportional to the existing sympathetic tone and the concentration of beta-adrenergic receptor blocking agent which reaches the receptor sites.
  • Examples of compounds which fit the definition of beta- adrenergic receptor blocking agent include but are not limited to timolol, sotalol, esmolol, cateolol, propranolol, betaxolol, penbutolol, metoprolol, acebutolol, atenolol, metoprolol, pindolol, and bisoprolol, and their salts, hydrates, solvates and any crystal forms in which they may occur.
  • Examples of compounds which fit the definition of selective IKs antagonists include, but are not limited to, the following:
  • A is 1) thieno
  • p is 0 or 1 ;
  • Ri is 1) phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02,
  • R2 is 1 ) phenyl, either unsubstituted or substituted with Cl-3 alkoxy or 4,4-dimethyloxazolin-2-yl,
  • Cl-6 alkyl either straight or branched chain, and either unsubstituted or substituted with Cl-3 alkoxy or Cl-3 alkoxy-Cl-3 alkoxy,
  • R2 is phenyl
  • the 2-position of the phenyl can be joined to the 4-position nitrogen of the diazepine ring through a carbonyl group and the double bond between the 4-nitrogen and the 5-carbon becomes a single bond
  • R is 1 ) hydrogen or
  • R4 is 1 ) hydrogen
  • Cl-6 alkyl the chain of carbon atoms of which can be interrupted by one or two non-adjacent oxygen atoms and which is either unsubstituted or substituted with Cl-3 alkoxycarbonyl, -OH or
  • R5 is hydrogen or oxygen or is joined to R2 to form the partial structure:
  • This invention is meant to include the individual diastereomers where such exist and mixtures thereof and enantiomers and mixtures of the enantiomers.
  • the pharmaceutically acceptable salts of the compounds of Formulas I include the conventional non-toxic salts or the quartemary ammonium salts of the compounds of Formula I 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-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like.
  • the pharmaceutically acceptable salts of the present invention can be synthesized from the compounds of Formula I which contain a basic or acidic moiety by conventional chemical methods. Generally, the salts are prepared by reacting the free base or acid with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid or base in a suitable solvent or various combinations of solvents.
  • A is benzo
  • X and Y are oxygen
  • R3 is methyl
  • R4 is hydrogen
  • R2 is Cl-6 alkyl.
  • X and Y are oxygen
  • R is methyl
  • R4 is hydrogen
  • R2 is phenyl
  • Z is Cl -6 alkylene or a bond
  • Ri is phenyl, phenyl substituted with -CI, -Br, -I, -F, or -CF3, or R is cyclohexyl.
  • Z is C2-4 alkenylene and R 1 is phenyl or phenyl substituted with -CI, -Br, -F, -I, -CF3, Cl-3 alkyl, Cl -3 alkoxy or methy lenedioxy.
  • a third embodiment of the compounds useful in the novel method of treatment of this invention is that wherein: Z is -NH-.
  • Another embodiment of this invention is a group of compounds, active in the novel method of treatment of this invention, which are novel compounds per se. These novel compounds are depicted in the following Table VI.
  • Another embodiment of this invention is a group of compounds which are active in the novel method of treatment of this invention. These compounds are depicted as follows:
  • X and Y are independently hydrogen, chloro, fluoro, bromo, iodo, or trifluoromethyl and
  • n 0, 1 or 2;
  • R is hydrogen, fluoro, chloro, bromo, iodo, or trifluoromethyl, methyl, or methoxy;
  • racemates mixtures of enantiomers, individual diastereomers or individual enantiomers with all isomeric forms and pharmaceutically acceptable salts, hydrates or crystal forms thereof, which are antiarrhythmic agents.
  • Yet another embodiment of this invention is a group of compounds which are active in the novel method of treatment of this invention. These compounds are depicted as follows:
  • R and R ⁇ are independently
  • phenyl either unsubstituted or substituted with one or two substituents selected from a) -N02, OH, b) -CI, Br, F, or I, c) -CF3, d) -Cl-3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy, and
  • Z is 1) Cl-6 alkyl, either straight or branched chain ,
  • the selective IKs blockers of the present invention have the pharmacological properties required for antiarrhythmic agents of Class III, namely they demonstrate prolongation of QTc-interval , and dose dependent increases in ventricular refractoriness. This is accomplished without effecting heart rate, mean arterial pressure and PR and QRS intervals. Modest increases in LV+dP/dt (left ventricular change in pressure with time) is observed. Further, these compounds suppress the induction of PVS (Programmed Ventricular Stimulation) induced ventricular tachy arrhythmias.
  • PVS Programmed Ventricular Stimulation
  • these compounds are effective in treating and preventing all types of arrhythmias including ventricular, atrial and supraventricular arrhythmias.
  • the compounds of the present invention are especially useful for controlling reentrant arrhythmias and prevent sudden death due to ventricular fibrillation. These compounds are also effective in treating and preventing impaired cardiac pump functions.
  • a selective IKs antagonist is administered in an amount ranging from about .0001 to about 10 mg per kg of body weight per day, preferably from about .0001 to about 2 mg per kg of body weight per day, and more preferably, when intravenous delivery of the compounds is employed, from about 0.0003 to about 0.3 mg per kg of body weight per day, or when given orally from about 0.01 to about 1 mg per kg of body weight per day, in a single dose or in 2 to 4 divided doses of each compound.
  • the beta-adrenergic receptor blocking agent is administered in an amount ranging from about 1 mg per day to about 300 mg poer day and more preferably from about 2 mg/day to about 250 mg per day.
  • the activity of the compounds described herein as antiarrhythmic agents is measured by their ability to block the IK and IKr currents as determined by the following test protocol.
  • Outward potassium currents are measured in single guinea pig ventricular myocytes using a whole-cell voltage clamp technique described in detail elsewhere (Sanguinetti and Jurkiewicz, 1990, Two components of cardiac delayed rectifier K+ current: differential sensitivity to block by Class LII antiarrhythmic agents. J. Gen Physiol. 96: 195-215).
  • Myocytes are isolated by enzymatic (collagenase and protease) digestion of Langandorf perfused hearts. Single cells are then voltage clamped using 1 mm square-bore pipettes filled with 0.5 M Kgluconate, 25 mM KC1, 5 mM K(2)ATP. Cells are bathed in a solution containing, in mN: 132 NaCl, 4KC1, 1.2 MgCl2, 10 HEPES, 10 glucose: pH 7.2, temp. 35°C.
  • Test depolarizations are applied as voltage ramps from -85 to -50 mV, and as steps to -10 mV (0.5 s) and +50 mV (1.0 s).
  • I ⁇ i is measured as peak outward current during the voltage ramp.
  • IKr is measured as tail currents upon repolarization from -10 mV to -50 mV.
  • IKs is measured as time-dependent current during the pulse to +50 mV. Currents are measured during control, then after exposure to drug at two different concentrations.
  • the compounds described herein as selective IKs blockers have an IC50 of less than 100 nM as IKs blockers.
  • the compounds of this invention are at least 10 times more potent in the blockade of IKs than of blockade of IKr-
  • Oxalyl chloride (158 mL, 230 mg, 1.81 mmol) was added to a mixture of 3-phenylpropanoic acid (249 mg, 1.66 mmol) and DMF (1 drop) in THF (10 mL) and the mixture was stirred at room temperature for 40 min.
  • 3(R)- Amino- 1 ,3-dihydro-l -methyl-5-phenyl-2H- 1 ,4- benzodiazepin-2-one J. Org. Chem. 1987, 52, 3232-3239
  • triethylamine (252 mL, 183 mg, 1.81 mmol) were added and the mixture was stirred at room temperature for 18 h.
  • (+)-N-[(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-v ⁇ -2.2-diphenylethanamide m.p. 200-201°C, [ ⁇ ]D +97.0° (c 0.168, CH2CI2).
  • the benzylamine (2.07 g, 7.9 mmol) was dissolved in methanol (60 mL), BOC2O (1.72 g, 7.9 mmol) added and the mixture hydrogenated at 50 psi over 10% palladium hydroxide on charcoal (200 mg) for 18 hours.
  • the reaction mixture was filtered through celite, washed with methanol and the filtrate evaporated to give 1 -t-butoxy- carbonylpiperidine-4,4-diethanol (2.0 g).
  • (+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l,4-benzo- diazepin-3-yl]-l '-(l ,1 -dimethylethoxycarbonyl)spiro(cyclohexan-4,4'- piperidineV 1 -carboxamide m.p. 135-138°C, [ ⁇ ]D +58.8° (C 0.925, CHCI3).
  • Bromoacetyl bromide (165 mL, 383 mg, 1.9 mmol) was added to an ice cooled solution of 3(R)-amino-l ,3-dihydro-l-methyl-5- phenyl-2H-l ,4-benzodiazepin-2-one (J. Org. Chem. 1987, 52, 3232- 3239) (500 mg, 1.88 mmol) and triethylamine (264 mL, 192 mg, 1.9 mmol) in methylene chloride (10 mL) and the mixture was stirred at room temperature for 1 h.
  • Phenol (104 mg, 1.1 mmol) was added to a suspension of sodium hydride (60% dispersion in mineral oil, 44 mg, 1.1 mmol) in toluene (10 mL).
  • sodium hydride 60% dispersion in mineral oil, 44 mg, 1.1 mmol
  • toluene 10 mL
  • N-[(3R)-2,3- dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzodiazepin-3-yl] -2-bromo- acetamide 400 mg, 1.04 mmol was added and the mixture was stirred at room temperature for 1 h.
  • the mixture was washed with water (3 x 15 mL), dried (MgS04) and the solvent was evaporated under reduced pressure.
  • 3-Bromopropionyl chloride (2.01 mL, 3.428 g, 20 mmol) was added to an ice cooled solution of 3(R)-amino-l ,3-dihydro-l - methyl-5-phenyl-2H-l ,4-benzodiazepin-2-one (J. Orb. Chem. 1987, 52, 3232-3239) (5.0 g, 18.8 mmol) and triethylamine (2.79 mL, 2.02 mg, 20 mmol) in methylene chloride (85mL) and the mixture was stirred at room temperature for 18 h.
  • 2,3-Dihydro-5 -phenyl- 1 H- 1 ,4-benzodiazepin-2-one (1.00 g, 4.23 mmol) was added to hexane washed sodium hydride (60% dispersion in mineral oil, 186 mg, 4.65 mmol) in DMF (5 mL). Further DMF (10 mL) was added and the mixture was stirred at room temperature. 2-(Dimethylamino)ethyl chloride hydrochloride (0.73 g, 5 mmol) was added to hexane washed sodium hydride (60% dispersion in mineral oil, 200 mg, 5.0 mmol) in DMF (5 mL) and the mixtures were combined.
  • Ethyl isocyanate (320 mL, 287 mg, 4.0 ⁇ unol) was added to a mixture of 2,3-dihydro-l -(2-dimethy laminoethy l)-3-hydroxyimino-5- phenyl-lH-l ,4-benzodiazepin-2-one (0.91 g, 2.7 mmol) and triethylamine (0.56 mL, 0.41 g, 4.0 mmol) in THF (30 mL). The mixture was heated under reflux for 7 h., further ethyl isocyanate (167 mL, 150 mg, 2.1 mmol) was added and the mixture was heated under reflux for 12 h.
  • Triethylamine was added to a mixture of 3-amino-2,3- dihydro- 1 -(2-dimethylaminoethyl)-5-phenyl- 1 H- 1 ,4-benzodiazepin-2- one (180 mg, 0.6 mmol), 3-(2,4-dichlorophenyl)propanoic acid (131 mg, 0.6 mmol), l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (115 mg, 0.6 mmol) and 1 -hydroxyberzotriazole (81 mg, 0.6 mmol) in DMF (15 mL) until the pH was 9.0. The mixture was stirred at room temperature for 72 h.
  • the layers were separated and the aqueous layer was extracted with ethyl acetate (5 mL). The combined organic fractions were washed with brine, dried (Na2S ⁇ 4) and the solvent was evaporated under reduced pressure. The residue was purified by flash column chroma-tography on silica gel, eluting with EtOAc/Hexane (65:35 increasing to 100:0). The first compound to elute was suspended in ethanol (1 mL) and ethanolic HC1 (6 M, 0.11 mL) was added. The mixture was stirred, then the solvent was evaporated under reduced pressure.
  • (+)-3(R)- ⁇ N-[3-(4-Chlorophenyl)prop- 1 -yl]amino ⁇ - 1 ,3-dihydro- 1 - methyl-5- phenyl-2H- 1.4-benzodiazepin-2-one hydrochloride m.p. 167-168°C, [ ⁇ ]D +20.8° (c 0.500, MeOH).
  • (+)-pheny Imethy 1 N-[(3R)-2,3-dihydro-l- methyl-5-phenyl-2-oxo-lH-l ,4-benzodiazepin-3-yl]carbamate (4.0 g, 10 mmol) and 2,4-bis(4-methoxyphenyl)-l ,3-dithia-2,4-diphosphetane-2,4- disulfide (4.5 g, 1 1 mmol) in toluene (100 mL) was heated under reflux for 75 min. The mixture was cooled and the volume was reduced to 30 mL by evaporation under reduced pressure.
  • Triethylamine (6.8 mL, 4.94 g, 49 mmol) was added to a heated (33°C) mixture of b-oxobenzenepropanenitrile (18.6 g, 128 mmol) and l,2-dithiane-2,5-diol (9.8 g, 64 mmol) in ethanol (120 mL) and the mixture was stirred at 50C° for 18 h. The mixture was cooled and the solvent was evaporated under reduced pressure. Dichloromethane was added, the mixture was washed with aqueous hydrochloric acid (0.5M), aqueous sodium hydroxide (IM) and brine, dried (Na2S ⁇ 4) and the solvent was evaporated under reduced pressure.
  • aqueous hydrochloric acid 0.5M
  • IM aqueous sodium hydroxide
  • brine dried (Na2S ⁇ 4)
  • 3-Methylphenylisocyanate 60 mL, 62 mg, 0.46 mmol was added to a solution of 3-amino-2,3-dihydro-l-methyl-5-phenyl-lH- thieno[2,3-e]-l,4-diazepin-2-one (124 mg, 0.46 mmol) in tetrahydro- furan (5 mL). The mixture was stirred at room temperature for 2 h. and the solvent was evaporated under reduced pressure.
  • Triethylamine (75 mL, 54 mg, 0.54 mmol) was added to a mixture of 3-amino-2,3-dihydro-l-methyl-5-phenyl lH-thieno[2,3-e]- 1 ,4-diazepin-2-one (82 mg, 0.3 mmol), cyclohexanepropanoic acid (52 mL, 47 mg, 0.3 mmol), l-(3-dimethylaminopropyl)-3-ethylcarbodi- imide hydrochloride (58 mg, 0.3 mmol) and 1-hydroxybenzotriazole (42 mg, 0.3 mmol) in DMF (1.5 mL).
  • Phenylmethyl N-[5-cyclohexyl-2,3-dihydro-2-oxo-lH-l ,4- benzodiazepin-3-yl]carbamate (150 mg, 0.38 mmol) was dissolved in hydrogen bromide in acetic acid (30%, 0.5 mL). After 2 h., ether was added and the solid was collected and dried in vacuo. THF (3 mL) and triethylamine (0.45 mL, 33 mg, 0.32 mmol) were added and the mixture was stirred at room temperature for 3 h.
  • Methanesulfonyl chloride (0.040 mL, 0.52 mmol) was added to a solution of (+)-N-[(3R)-7-amino-2,3-dihydro-l-methyl-2- oxo-5-phenyl-lH-l,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)- propanamide (193 mg, 0.40 mmol) and pyridine (0.065 mL, 0.80 mmol) in methylene chloride (1.6 mL). The resulting solution was stirred 2 h.
  • the solution was diluted with ethyl acetate (12 mL), washed with IN HC1, water, saturated sodium bicarbonate solution, water, and brine (3 mL each), dried (Na2S04) and the solvent was evaporated under reduced pressure. The residue was dissolved in warm toluene, treated with charcoal, and filtered.
  • the mixture was stirred at -20°C for 30 min., then poured into a mixture of water (50 mL), acetic acid (3 mL), and ethyl acetate (65 mL). The mixture was stirred to dissolve all solids and the layers were separated. The aqueous layer was extracted with ethyl acetate (65 mL). The combined organic fractions were washed with saturated sodium bicarbonate solution and brine (20 mL each), dried (Na2S04), and the solvent was evaporated under reduced pressure.
  • the residue was purified by preparative plate chromatography on silica gel eluting with methanol/ chloroform/acetic acid (5:95:1).
  • the purified material was stirred under chloroform (5 mL) with potassium carbonate (0.1 g) and water (2 drops) for 5 min.
  • the mixture was dried (Na2S ⁇ 4) and the solvent was evaporated under reduced pressure.
  • the residue was suspended in ethanol (2 mL) and ethanolic HC1 (6.8 M, 0.147 mL) was added.
  • the mixture was stirred at -20°C for 30 min., then poured into a mixture of water (25 mL), acetic acid (2.5 mL), and ethyl acetate (55 mL). The mixture was stirred to dissolve all solids and the layers were separated. The aqueous layer was extracted with ethyl acetate (2 x 55 mL). The combined organic fractions were washed with saturated sodium bicarbonate solution and brine (20 mL each), dried (Na2S ⁇ 4), and the solvent was evaporated under reduced pressure.
  • the benzodiazepine obtained in Step C was converted to the oxime as described in Example 80 Step A.
  • the oxime (2 gms) was dissolved in acetic acid (150 mL) and 10% Pd/C (1 gm) added. The mixture was stirred rapidly under an atmosphere of hydrogen for 90 min or until complete by HPLC. The reaction was filtered, the catalyst washed with methylene chloride (200 mL) and the filtrates concentrated in vacuo to an oil. The oil was dissolved in saturated aqueous sodium bicarbonate (100 mL) and product extracted with ethyl acetate (3 x 150 mLs). Concentration of the dried (Na2S ⁇ 4) extracts gave 2.60 gms (97%).
  • Step F The anine was coupled with 3-(2,4-dichlorophenyl)- propionic acid as described in Example 43 to yield N-(2,3-dihydro-l- methyl-2-oxo-5-isopropyl-lH-l ,4-benzodiazepin-3-yl)-3-(2,4- dichlorophenyl)propanamide.
  • Step A Preparation of N-tert-butyloxycarbonyl-4-(4-chloro- benzyl)-4-piperidinecarboxylic acid
  • N-Boc-ethylisonipecotate (51.4 g, 200 mmole) in THF (1L) at -60° C was treated with a solution of lithium bistrimethylsilyl amide (220 mL of a 1 N solution in THF, 220 mmole). After stirring at -60°C for 5 minutes, a solution of 4-chlorobenzyl chloride (33.8 g, 210 mmole) in THF (200 mL) was added and the reaction allowed to warm to room temperature. Most of the THF (about 800 mL) was removed by evaporation at reduced pressure. The remainder was poured into 1 L of 1 N HCl and extracted with two 800 mL portions of ethyl acetate.
  • Step B Preparation of N-( 1 ,3-dihydro- 1 -methyl-2-oxo-5-phenyl- 2H- 1 ,4-benzodiazepin-3-yl)-4-(4-chlorobenzyl)-4-piper- idinecarboxamide dihydrochloride
  • (+)-3-Cyclohexyl-N-[2,3-dihydro- 1 -methyl-2-oxo-5- phenyl-lH-l ,4-benzodiazepin-3-yl]propanamide (2.0 g, 5.0 mmol) was dissolved in tetrahydrofuran (30 mL), cooled to 0°C and methyl magnesium chloride (3M, 2.0 mL) was added. After 0.25 h, paraformadehyde (0.15 g, 10 mmol) was added, and the mixture was allowed to warm to room temperature. The reaction was then diluted with ethyl acetate (150 mL) and saturated aqueous sodium hydrogen carbonate (150 mL) was added.
  • (+)-3-Cyclohexyl-N-[2,3-dihydro- 1 -methyl-2-oxo-5- phenyl-lH-l ,4-benzodiazepin-3-yl]-N-(hydroxymethyl)propanamide (0.67 g, 1.56 mmol) was dissolved in methylene chloride(100 mL), along with tetrazole (0.33 g, 4.7 mmol), and then N,N-diisopropyl- dibenzyl-phosphoramidite (1.07 g, 3.1 mmol). After 2 h, the mixture was diluted with methylene choride (150 mL), and extracted with saturated aqueous sodium hydrogen carbonate (3 x 100 mL).
  • Step A Preparation of 2,3-dihydro-2-oxo-5-phenyl-l -(2,2,2- trifluoroethyl)- 1 H-benzo[e] [ 1 ,4]diazepine.
  • Step C Preparation of racemic 3-Amino-5-phenyl-l -(2,2,2- trifluoroethyl)- 1 H-benzo[e] [ 1 ,4]diazepine.
  • Step D Preparation of 2-Amino-N-[2-oxo-5-phenyl- 1 -(2,2,2- trifluoroethyl)-2,3 -dihydro- 1 H-benzo[e] [ 1 ,4]diaze ⁇ in- 3-yl]-3-phenylpropionamide
  • Step E Preparation of 3(R)-(+)-3-Amino-5-phenyl- 1 -(2,2,2- trifluoroethyl)-l H-benzo[e] [1 ,4] diazepine.
  • Step F Preparation of (+)-3,5-Dichloro-N-[3R-2,3-dihydro-2-oxo-
  • (+)-3R-3-amino-5-phenyl-l- (2,2,2-trifluoroethyl)-lH-benzo[e][ l,4] diazepine (5.6 g, 16.8 mmol) in DMF (50 mL) was added l-(3-Dimethylaminopropyl-3-ethylcarbodi- imide hydrochloride(4.44 g, 23.0 mmol), and 1-hydroxybenztriazole hydrate (3.11 g, 23.0 mmol) and 3,5-Dichlorobenzoic acid (3.21 g, 16.8 mmol). This was stined at ambient temperature for 2 hours.
  • the reaction was diluted with 500 mL satd. NaHC ⁇ 3 and extracted with 2 x 300 mL ethyl acetate. The combined organics were washed with 10% KHSO4 (200 mL) , brine (200 mL), dried over Na2S04, and evaporated to a white foam. This was chromatographed over a 75 x 200 mm silica column eluting with 20% ethyl acetate:hexane. The pure fractions were collected and evaporated under reduced pressure to give 8.5 g of a white foam which was crystallized from 15% ethyl acetate :hexane to give 5.3 g of a white powder .

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Epidemiology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Plural Heterocyclic Compounds (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
  • Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)

Abstract

A method of preventing, treating, terminating and protecting against cardiac arrhythmias, such as atrial, supraventricular and ventricular ectopy, tachycardia, flutter or fibrillation, including atrial, supraventricular and ventricular arrhythmias resulting from myocardial ischemic injury in a patient in need thereof, comprising administration of a selective IKs antagonist and a beta-adrenergic receptor blocking agent, administered in combined therapy either simultaneously, separately or sequentially is presented. Additionally, a pharmaceutical preparation comprising a selective IKs antagonist and a beta-adrenergic receptor blocking agent, wherein these compounds are administered simultaneously, separately or sequentially is presented.

Description

TITLE
PHARMACEUTICAL PREPARATION
BACKGROUND OF THE INVENTION
The present invention relates to the co-administration, either simultaneously, separately or sequentially of a selective iKs antagonist and a beta-adrenergic receptor blocking agent for use in preventing, treating and terminating cardiac arrhythmias, such as atrial, supraventricular and ventricular ectopy, tachycardia, flutter or fibrillation, including atrial, supraventricular and ventricular arrhythmias resulting from myocardial ischemic injury in a patient in need thereof. This invention also relates to a pharmaceutical formulation which comprises a selective IKs antagonist and a beta- adrenergic receptor blocking agent along with a pharmaceutically acceptable carrier.
Arrhythmias often occur as complications to cardiac diseases such as myocardial infarction and heart failure. In a serious case, arrhythmias give rise to ventricular fibrillation and can cause sudden death.
Though various antiarrhythmic agents are now available on the market, agents exhibiting both satisfactory effects and high safety profiles, are not yet available for patients. For example, antiarrhythmic agents of Class I, according to the classification of Vaughan-Williams, which cause a selective inhibition of the maximum velocity of the upstroke of the action potential (Vmax) are inadequate for preventing ventricular fibrillation. In addition, they have problems regarding safety, namely, they cause a depression of the myocardial contractility and have a tendency to induce arrhythmias due to an inhibition of the impulse conduction. Beta-adrenergic receptor blocking agent which belong to Class II are of limited value since their effects are either limited to a certain type of arrhythmia or are contraindicated because of their cardiac depressant properties in certain patients with cardiovascular disease. Their safety, however, is higher than that of the antiarrhythmic agents of Class I. Antiarrhythmic agents of Class III are drugs which cause a selective prolongation of the duration of the action potential without a significant depression of the Vmax. Until recently, drugs in this class were limited to sotalol and amiodarone, both of which have been shown to possess Class IH properties. However, Sotalol also possesses Class II effects which may cause cardiac depression and be contraindicated in certain susceptible patients. Amiodarone is severely limited by side effects. Drugs of this class are expected to be effective in preventing ventricular fibrillations. Pure Class UI agents, by definition, are not considered to cause myocardial depression or an induction of arrhythmias due to the inhibition of the action potential conduction as seen with Class I antiarrhythmic agents.
Recently, a novel group of Class BT agents have been disclosed which antagonize the IKs channel found in heart muscle. These compounds IKs channel antagonists are effective in treating and preventing all types of arrhythmias including ventricular and atrial (supraventricular) arrhythmias. These novel compounds are disclosed and claimed in U.S. Patent Application, Serial Nos. 08/411,240; 08/516,467; and 08/516,226 which are hereby incoφorated by reference. These novel compounds are especially useful for controlling reentrant arrhythmias and preventing sudden death due to ventricular fibrillation. These compounds are also effective in treating and preventing impaired cardiac pump functions.
In the treatment of arrhythmia, IKs antagonists have demonstrated effectiveness when delivered orally in amounts ranging from about 0.01 to about 1 mg per kg of body weight per day, in a single dose or in 2 to 4 divided doses.
The activity of the compounds described herein as antiarrhythmic agents is measured by their ability to block the IKs and iKr currents as determined by the following test protocol.
Outward potassium currents are measured in single guinea pig ventricular myocytes using a whole-cell voltage clamp technique described in detail elsewhere (Sanguinetti and Jurkiewicz, 1990, Two conn ents of cardiac delayed rectifier K+ current: differential sensitivity to block by Class LU antiarrhythmic agents. J. Gen Physiol. 96: 195-215). Myocytes are isolated by enzymatic (collagenase and protease) digestion of Langandorf perfused hearts. Single cells are then voltage clamped using 1 mm square-bore pipettes filled with 0.5 M Kgluconate, 25 mM KC1, 5 mM K(2)ATP. Cells are bathed in a solution containing, in mN: 132 NaCl, 4KC1, 1.2 MgCl2, 10 HEPES, 10 glucose: pH 7.2, temp. 35°C.
Each cell is maintained at a holding potential of -50 mV. Test depolarizations are applied as voltage ramps from -85 to -50 mV, and as steps to -10 mV (0.5 s) and +50 mV (1.0 s). Iκi is measured as peak outward current during the voltage ramp. iKr is measured as tail currents upon repolarization from -10 mV to -50 mV. IKs is measured as time-dependent current during the pulse to +50 mV. Currents are measured during control, then after exposure to drug at two different concentrations.
Employing this test the compounds described herein as selective IKs channel antagonists, have an IC50 of less than 100 nM as IKs antagonists. The compounds of this invention are at least 10 times more potent in the blockade of IKs than of blockade of IKr-
Beta-adrenergic receptor blocking agents, or "beta- blockers", are a class of pharmaceutically active compounds which decrease the positive chronotropic, positive inotropic, bronchodilator and vasodilator responses caused by beta-adrenergic receptor agonists. The magnitude of this decreased response is proportional to the existing sympathetic tone and the concentration of beta-blocker at the receptor sites. Beta-adrenergic receptor blockage is said to reduce cardiac output in both healthy subjects and patients with heart disease. While the mechanism of antihypertension effects of beta-adrenergic receptor blocking agents has not been established, possible mechanisms of action include reduction in cardiac output, reduction in plasma renin activity, and central nervous system sympatholytic action. The administration of beta-adrenergic receptor antagonists has been shown effective in reducing the incidence of mortality and sudden death in postinfarction patients (Yusaf et al., Prog Cardiovasc Dis 17: 335-371, 1985; Lau et al., N Eng J Med 327: 248-254, 1992).
While both selective IKs channel blockers and beta- adrenergic receptor blocking agents have been proven effective when administered separately, it is considered to be in the best interest of the patient to reduce the amount of these compounds provided to the patient. Any reduction of one or the other compound would be considered helpful, but this is particularly true of beta-adrenergic receptor blocking agents which are known to have significant side effects in some humans.
SUMMARY OF THE INVENTION
A method is presented for use in preventing, treating and terminating cardiac arrhythmias, such as atrial, supraventricular and ventricular ectopy, tachycardia, flutter or fibrillation, including atrial, supraventricular and ventricular arrhythmias resulting from myocardial ischemic injury in a patient in need thereof which comprises the co- administration, either simultaneously, separately or sequentially of a selective IKs antagonist and a beta-adrenergic receptor blocking agent. This invention also relates to a pharmaceutical formulation which comprises a selective IKs antagonist and a beta-adrenergic receptor blocking agent along with a pharmaceutically acceptable carrier.
DETAILED DESCRIPTION OF THE INVENTION
A method is presented for use in preventing, treating and terminating cardiac arrhythmias, such as atrial, supraventricular and ventricular ectopy, tachycardia, flutter or fibrillation, including atrial, supraventricular and ventricular arrhythmias resulting from myocardial ischemic injury in a patient in need thereof which comprises the co- administration, either simultaneously, separately or sequentially of a selective IKs antagonist and a beta-adrenergic receptor blocking agent. This invention also relates to a pharmaceutical formulation which comprises a selective IKs antagonist and a beta-adrenergic receptor blocking agent along with a pharmaceutically acceptable carrier. By a "selective IKs antagonist" is meant those compounds which when studied in the test disclosed above have an IC50 of less than 100 nM as IKs blockers. The compounds of this invention are at least 10 times more potent in the blockade of IKs than of blockade of IKr-
Beta-adrenergic receptor blocking agents are compounds which decrease the positive chronotropic, positive inotropic, bronch- odilator and vasodilator responses caused by beta-adrenergic receptor agonists. The magnitude of this decreased response is proportional to the existing sympathetic tone and the concentration of beta-adrenergic receptor blocking agent which reaches the receptor sites.
Examples of compounds which fit the definition of beta- adrenergic receptor blocking agent include but are not limited to timolol, sotalol, esmolol, cateolol, propranolol, betaxolol, penbutolol, metoprolol, acebutolol, atenolol, metoprolol, pindolol, and bisoprolol, and their salts, hydrates, solvates and any crystal forms in which they may occur.
Examples of compounds which fit the definition of selective IKs antagonists include, but are not limited to, the following:
Figure imgf000007_0001
or a pharmaceutically acceptable salt thereof, wherein
A is 1) thieno,
2) pyrido, or
3) benzo either unsubstituted or substituted with -NH2 -NHS02 (C1-3 alkyl), Cl-3 alkyl or C 1 -3 alkoxy;
Figure imgf000008_0001
Z i 1 ) Cl-6 alkylene, either straight or branch chain and either unsubstituted or substituted with phenyl or spiro-piperidine,
2) C2-4 alkenylene, either straight or branch chain,
3) -(CH2)m-W-(CH2)n- wherein m and n are independently 0, 1, 2, 3 or 4 and W is -0-, -S- or -NH,
4) 4-(5-methylisoxazole-3-yl),
5) C3-6 cycloalkylene, or
6) single bond;
p is 0 or 1 ;
Ri is 1) phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02,
Figure imgf000008_0002
4) mono- or bicyclic heterocyclyl of 5 to 10 members one or two of which are sulfur, nitrogen or oxygen, the remaining being carbon, such as 2-thienyl, 2-furanyl, 2-indolyl, 2- quinoxolinyl, or 2-(2,3-dihydro benzofuranyl)
5) Cl-3 alkyl, or
6) indan-5-yl;
R2 is 1 ) phenyl, either unsubstituted or substituted with Cl-3 alkoxy or 4,4-dimethyloxazolin-2-yl,
2) Cl-6 alkyl, either straight or branched chain, and either unsubstituted or substituted with Cl-3 alkoxy or Cl-3 alkoxy-Cl-3 alkoxy,
3) C5-7 cycloalkyl,
4) 2- or 3-furyl,
5) l-methylpiperidin-2-yl, or
6) if R2 is phenyl, the 2-position of the phenyl can be joined to the 4-position nitrogen of the diazepine ring through a carbonyl group and the double bond between the 4-nitrogen and the 5-carbon becomes a single bond;
R is 1 ) hydrogen or
2) Cl-3 alkyl either unsubstituted or substituted with -N(CH3)2, -OH, -CF3, or
3) -CF3;
R4 is 1 ) hydrogen,
2) Cl-6 alkyl, the chain of carbon atoms of which can be interrupted by one or two non-adjacent oxygen atoms and which is either unsubstituted or substituted with Cl-3 alkoxycarbonyl, -OH or
Figure imgf000009_0001
3) tetrazol-5-yl; R5 is hydrogen or oxygen or is joined to R2 to form the partial structure:
Figure imgf000010_0001
the bond represented by is:
1 ) a double bond when p is zero or when p is 1 and R^ is oxygen, or
2) a single bond when R^ is hydrogen or R^ is joined to R^ to form the partial structure:
Figure imgf000010_0002
This invention is meant to include the individual diastereomers where such exist and mixtures thereof and enantiomers and mixtures of the enantiomers.
The pharmaceutically acceptable salts of the compounds of Formulas I include the conventional non-toxic salts or the quartemary ammonium salts of the compounds of Formula I formed, e.g., from non-toxic inorganic or organic acids. For example, 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-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like.
The pharmaceutically acceptable salts of the present invention can be synthesized from the compounds of Formula I which contain a basic or acidic moiety by conventional chemical methods. Generally, the salts are prepared by reacting the free base or acid with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid or base in a suitable solvent or various combinations of solvents.
One embodiment of this invention are novel compounds useful in the novel method of treatment of this invention wherein:
A is benzo;
X and Y are oxygen;
R3 is methyl;
R4 is hydrogen; and
R2 is Cl-6 alkyl.
Specific novel compounds representative of this embodiment are those of the following structure and specified in Table I:
TABLE I
Figure imgf000011_0001
Rl R2
Figure imgf000012_0002
Another embodiment of the compounds useful in the novel method of treatment of this invention is that wherein:
A is
Figure imgf000012_0001
X and Y are oxygen;
R is methyl;
R4 is hydrogen; and
R2 is phenyl.
A class of novel compounds within this embodiment is that with structural formula:
Figure imgf000013_0001
wherein
Z is Cl -6 alkylene or a bond and
Ri is phenyl, phenyl substituted with -CI, -Br, -I, -F, or -CF3, or R is cyclohexyl.
Specific novel compounds representative of this class are those depicted in the following Table II:
Figure imgf000013_0002
TABLE II (Cont'd)
Figure imgf000014_0002
Another class of novel compounds within this embodiment is that with structural formula:
Figure imgf000014_0001
wherein Z is C2-4 alkenylene and R 1 is phenyl or phenyl substituted with -CI, -Br, -F, -I, -CF3, Cl-3 alkyl, Cl -3 alkoxy or methy lenedioxy.
Specific novel compounds representative of this class are those depicted in the following Table ID:
Figure imgf000015_0001
TABLE III (Cont'd)
-CH=CH- 4-IPh
-CH=CH- 4-BrPh
-C=CH-
I Ph
CH3
-CH=CH- Ph'
-CH=CH- 3,4-diCIPh
-CH=CH- 4-CH3Ph
-CH=CH- 4-CH3OPh
-CH=CH- 3,4-methylenedioxyPh
-CH=CH- 3-BrPh
*This compound is disclosed in U.S. Patent 4,820,834
A third embodiment of the compounds useful in the novel method of treatment of this invention is that wherein: Z is -NH-.
Compounds representative of this embodiment are those disclosed in the following Table IV. TABLE IV
Figure imgf000017_0001
R1 R' R^
Figure imgf000017_0002
benzo 3-CH3Ph Ph -CH, O
benzo 5-indanyl Ph ■HTT O
OH
\ Y^ 3-CH3Ph Ph -CH3 o
Other specific compounds included within the broadest genus but not included in one of the embodiments previously described are as shown in Table V. TABLE V
Figure imgf000018_0001
Representative of compounds wherein p is 1 is the compound of structural formula:
Figure imgf000018_0002
Representative of compounds wherein the bond between the 4 and 5 positions is a single bond is the compound of structural formula:
Figure imgf000018_0003
Representative of compounds wherein the bond represents a single bond and R^ is joined to R2 is the compound of structural formula:
Figure imgf000019_0001
Another embodiment of this invention is a group of compounds, active in the novel method of treatment of this invention, which are novel compounds per se. These novel compounds are depicted in the following Table VI.
Figure imgf000019_0002
Another embodiment of this invention is a group of compounds which are active in the novel method of treatment of this invention. These compounds are depicted as follows:
Figure imgf000020_0001
where
X and Y are independently hydrogen, chloro, fluoro, bromo, iodo, or trifluoromethyl and
n is 0, 1 or 2;
R is hydrogen, fluoro, chloro, bromo, iodo, or trifluoromethyl, methyl, or methoxy; and
the racemates, mixtures of enantiomers, individual diastereomers or individual enantiomers with all isomeric forms and pharmaceutically acceptable salts, hydrates or crystal forms thereof, which are antiarrhythmic agents.
Yet another embodiment of this invention is a group of compounds which are active in the novel method of treatment of this invention. These compounds are depicted as follows:
Figure imgf000020_0002
R and R^ are independently
1) phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02, OH, b) -CI, Br, F, or I, c) -CF3, d) -Cl-3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy, and
Z is 1) Cl-6 alkyl, either straight or branched chain ,
2) substituted Cl-6 alkyl, either straight or branched chain, wherein the substituents are selected from F, OH, Nθ2,
3) C2-4 alkenylene, either straight or branched chain,
4) -(CH2)m-W-(CH2)n- wherein m and n are independently 0, 1, 2, 3 or 4 and W is -0-, -S- or -NH,
5) C3-6 cycloalkane,
6) C3-6 cycloalkylene, or
7) single bond;
The selective IKs blockers of the present invention have the pharmacological properties required for antiarrhythmic agents of Class III, namely they demonstrate prolongation of QTc-interval , and dose dependent increases in ventricular refractoriness. This is accomplished without effecting heart rate, mean arterial pressure and PR and QRS intervals. Modest increases in LV+dP/dt (left ventricular change in pressure with time) is observed. Further, these compounds suppress the induction of PVS (Programmed Ventricular Stimulation) induced ventricular tachy arrhythmias.
Individually, these compounds are effective in treating and preventing all types of arrhythmias including ventricular, atrial and supraventricular arrhythmias. The compounds of the present invention are especially useful for controlling reentrant arrhythmias and prevent sudden death due to ventricular fibrillation. These compounds are also effective in treating and preventing impaired cardiac pump functions.
In the novel method of this invention of treating arrhythmia, a selective IKs antagonist is administered in an amount ranging from about .0001 to about 10 mg per kg of body weight per day, preferably from about .0001 to about 2 mg per kg of body weight per day, and more preferably, when intravenous delivery of the compounds is employed, from about 0.0003 to about 0.3 mg per kg of body weight per day, or when given orally from about 0.01 to about 1 mg per kg of body weight per day, in a single dose or in 2 to 4 divided doses of each compound. The beta-adrenergic receptor blocking agent is administered in an amount ranging from about 1 mg per day to about 300 mg poer day and more preferably from about 2 mg/day to about 250 mg per day.
The activity of the compounds described herein as antiarrhythmic agents is measured by their ability to block the IK and IKr currents as determined by the following test protocol.
Outward potassium currents are measured in single guinea pig ventricular myocytes using a whole-cell voltage clamp technique described in detail elsewhere (Sanguinetti and Jurkiewicz, 1990, Two components of cardiac delayed rectifier K+ current: differential sensitivity to block by Class LII antiarrhythmic agents. J. Gen Physiol. 96: 195-215). Myocytes are isolated by enzymatic (collagenase and protease) digestion of Langandorf perfused hearts. Single cells are then voltage clamped using 1 mm square-bore pipettes filled with 0.5 M Kgluconate, 25 mM KC1, 5 mM K(2)ATP. Cells are bathed in a solution containing, in mN: 132 NaCl, 4KC1, 1.2 MgCl2, 10 HEPES, 10 glucose: pH 7.2, temp. 35°C.
Each cell is maintained at a holding potential of -50 mV. Test depolarizations are applied as voltage ramps from -85 to -50 mV, and as steps to -10 mV (0.5 s) and +50 mV (1.0 s). Iκi is measured as peak outward current during the voltage ramp. IKr is measured as tail currents upon repolarization from -10 mV to -50 mV. IKs is measured as time-dependent current during the pulse to +50 mV. Currents are measured during control, then after exposure to drug at two different concentrations.
Employing this test the compounds described herein as selective IKs blockers have an IC50 of less than 100 nM as IKs blockers. The compounds of this invention are at least 10 times more potent in the blockade of IKs than of blockade of IKr-
Typical synthetic schemes employed in making the compounds herein are illustrated below.
SCHEME 1
Figure imgf000023_0001
SCHEME2
Figure imgf000024_0001
Figure imgf000024_0002
SCHEME3
Figure imgf000025_0001
2. TiCI3
3. Chromatography
CH3S02CI
Figure imgf000025_0002
SCHEME4
Figure imgf000026_0001
SCHEME 5
Figure imgf000026_0003
3. PhCON e2
Figure imgf000026_0002
Pyridine
Figure imgf000026_0005
Figure imgf000026_0006
N^^\ SCHEME6
Figure imgf000027_0001
SCHEME7
N- \
Figure imgf000028_0002
SCHEME8
R1NC0
Figure imgf000029_0001
Figure imgf000029_0002
SCHEME9
Figure imgf000030_0002
Figure imgf000030_0001
SCHEME O
GLY Ethyl ester NaH
Figure imgf000031_0003
Figure imgf000031_0001
Figure imgf000031_0004
Figure imgf000031_0002
Figure imgf000031_0005
SCHEMEU
Figure imgf000032_0001
Figure imgf000032_0002
CH2CI2, 0°C
Figure imgf000032_0003
SCHEME 12
Figure imgf000033_0001
SCHEME 13
Figure imgf000034_0001
R' R<
SCHEME 14
Figure imgf000035_0001
CH,
EDC, HOBT, TEA R1-Z-COOH
Figure imgf000035_0002
CH,
SCHEME 15
Figure imgf000035_0003
SCHEME 16
Figure imgf000036_0001
Figure imgf000036_0003
Figure imgf000036_0002
Figure imgf000036_0004
Figure imgf000037_0001
Figure imgf000037_0005
Figure imgf000037_0003
Figure imgf000037_0002
R-(CH2)nCOCI or R-(CH2)nC02H, EDC, HOBT
Figure imgf000037_0004
EXAMPLES
EXAMPLE 1
Figure imgf000038_0001
(E)-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yll-3-phenyl-2-propenamide
A solution of (E)-3-phenyl-2-propenoyl chloride (367 mg,
2.2 mmol) in methylene chloride (1 mL) was added to a solution of 3(R)-amino-l ,3-dihydro-l -methyl-5-phenyl-2H-l ,4-benzodiazepin-2- one (J. Org. Chem. 1987, 52, 3232-3239) (531 mg, 2.0 mmol) and triethylamine (307 mL, 225 mg, 2.2 mmol) in methylene chloride (10 mL). The mixture was stirred at room temperature for 25 min. and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2Cl2/Et2θ (95:5) and the residue was triturated with Et2θ. The solid was collected and dried in vacuo at 70°C to give (E)-(+)-N-[(3R)- 2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzodiazepin-3-yl]-3- phenyl-2-propenamide as a colorless solid (170 mg, 21 %), m.p. 140- 142°C, [α]D +86.7° (c=0.173, CH2CI2). dH (CDC13) 7.70-7.26 (16H, m), 6.63 (IH, d, J 15.6 Hz), 5.68 (IH, d, J
8.3 Hz), and 3.50 (3H, s).
Anal. Calcd. for C25H21N3O2.O. I5 (C2H5)2θ:
C, 75.63; H, 5.58; N, 10.33. Found: C, 75.29; H, 5.57; N, 10.33%. Employing the procedure substantially as described above, but substituting an appropriate acid chloride for the (E)-3-phenyl-2- propenoyl chloride, the following compounds were prepared:
EXAMPLE 2
Figure imgf000039_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin- 3 - yll benzamide m.p. 224-225°C, [α]D +89.2° (c = 0.141 , CH2CI2). dH (CDCI3) 8.04 (IH, d, J 8.1 Hz), 7.96 (2H, d, J 6.8 Hz), 7.64-7.36
(10H, m), 7.27 (2H, t, J 7.6 Hz), 5.74 (IH, d, J 7.8 Hz), and 3.51 (3H, s).
Anal. Calcd. for C23H19N3O2.O.2OH2O:
C, 74.06; H, 5.24; N, 11.26. Found: C, 74.13; H, 5.12; N, 1 1.16%.
EXAMPLE 3
Figure imgf000039_0002
First diastereoisomer to elute:
(-)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4- benzo-diazepin-3-yll(trans-2-phenyl- 1 -cyclopropane )carboxamide m.p. 180-181°C, [α]D -155.8° ( c = 0.434, CH2CI2). dH (CDCI3) 7.62-7.09 (15H, m), 5.59 (IH, d, J 8.1 Hz), 3.47 (3H, s), 2.52-2.45 (IH, m), 1.90-1.84 (IH, m),1.69-1.56 (IH, m), and 1.38-1.32 (IH, m).
Anal. Calcd. for 26H23N3O2.0.25H2O: C, 75.43; H, 5.72; N, 10.15. Found: C, 75.38; H, 5.64; N, 9.94%.
Second diastereoisomer to elute:
(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4- benzo-diazepin-3-yll(trans-2-phenyl- 1 -cvclopropanekarboxamide m.p. 104-107°C, [ ]D +328.2° (c = 0.098, CH2CI2). dH (CDCI3) 7.62-7.13 (15H, m), 5.60 (IH, d, J 8.3 Hz), 3.48 (3H, s),
2.59-2.54 (IH, m), 1.93-1.87 (IH, m),1.62-1.56 (IH, m, overlaps with water), and 1.33-1.25 (IH, m).
Anal. Calcd. for C26H23N3θ2.0.50H2θ.0.45PhCH3:
C, 76.13; H, 5.95; N, 9.14. Found: C, 76.10; H, 5.94; N, 9.17%.
EXAMPLE 4
Figure imgf000040_0001
(+)-N- [(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-pheny 1- 1 H- 1 ,4- benzodiazepin-3-yll-lH-indole-2-carboxamide m.p. 167-177°C, [α]D +1 13° (c = 1.103, CH2CI2). dH (CDCI3) 9.15 (IH, br s), 8.10 (IH, d, J 9.0 Hz), 7.75-7.10 (14H, m),
5.75 (IH, d, J 9.0 Hz), and 3.50 (3H, s).
Anal. Calcd. for C25H20N4O2:
C, 73.51; H, 4.94; N, 13.72. Found: C, 73.31; H, 4.80; N, 13.62%.
EXAMPLE 5
Figure imgf000041_0001
(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l,4-benzo- diazepin-3-yl1heptanamide m.p. 49-54°C, [α]D +69.5° (c=1.000, MeOH). Anal. Calcd. for C23H27N3O2.O.4OH2O: C, 71.81; H, 7.28; N, 10.92. Found: C, 71.90; H, 7.09; N, 10.85%.
EXAMPLE 6
Figure imgf000041_0002
(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l,4-benzo- diazepin-3-yllhexanamide
[α]D +72.6° (c=0.920, MeOH). Anal. Calcd. for C22H25N3O2:
C, 72.70; H, 6.93; N, 11.56. Found: C, 72.44; H, 6.75; N, 11.25%.
EXAMPLE 7
Figure imgf000042_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yllpentanamide
[ ]D +68.2° (c=1.310, MeOH).
Anal. Calcd. for C21H23N3O2.O.25CHCI3:
C, 68.21 ; H, 6.26; N, 11.26. Found: C, 68.2; H, 6.29; N, 11.17%.
EXAMPLE 8
Figure imgf000043_0001
(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5 -phenyl- lH-l ,4-benzo- diazepin-3-yll-3-phenylpropanamide
Oxalyl chloride (158 mL, 230 mg, 1.81 mmol) was added to a mixture of 3-phenylpropanoic acid (249 mg, 1.66 mmol) and DMF (1 drop) in THF (10 mL) and the mixture was stirred at room temperature for 40 min. 3(R)- Amino- 1 ,3-dihydro-l -methyl-5-phenyl-2H- 1 ,4- benzodiazepin-2-one (J. Org. Chem. 1987, 52, 3232-3239) (400 mg, 1.51 mmol) and triethylamine (252 mL, 183 mg, 1.81 mmol) were added and the mixture was stirred at room temperature for 18 h. The mixture was poured into saturated aqueous sodium hydrogen carbonate (20 mL) and extracted with ethyl acetate (3 x 20 mL). The combined organic fractions were dried (Na2Sθ4) and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2θ2/Et2θ (95:5) and the residue was recrystallized from toluene/hexane to give (+)-N-[(3R)-2,3- dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzodiazepin-3-yl]-3- phenylpropanamide as a colorless solid (380 mg, 63%), m.p. 179°C, [α]D +100.4° (c = 0.225, CH2CI2). dH (CDCI3) 7.62-7.57 (2H, m), 7.47-7.21 (13H, m), 5.54 (IH, d, J 8.1 Hz), 3.47 (3H, s), 3.03 (2H, t, J 7.8 Hz), and 2.73-2.67 (2H, m). Anal. Calcd. for C25H23N3O2.O.I5H2O:
C, 75.04; H, 5.87; N, 10.50. Found: C, 75.06; H, 5.78; N, 10.55%. Employing the procedure substantially as described above, but substituting an appropriate carboxylic acid for the 3-phenylpropanoic acid, the following compounds were prepared:
EXAMPLE 9
Figure imgf000044_0001
E-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yll-3-(3.4-dichlorophenylV2-propenamide m.p. 145-147°C, [α]D +77.8° (c=0.126, CH2CI2). dH (CDCI3) 7.64-7.25 (14H, m), 6.61 (IH, d, J 15.6 Hz), 5.65 (IH, d, J
8.0 Hz), and 3.50 (3H,.s).
Anal. Calcd. for C25H19N3O2CI2:
C, 64.67; H, 4.12; N, 9.05. Found: C, 64.57; H, 4.25; N, 9.01 %.
EXAMPLE 10
Figure imgf000044_0002
E-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yll3-(4-nitrophenyl)-2-propenamide m.p. 165-166 °C, [αjD +80.5° (c=0.126, CH2CI2). dH (CDC13) 8.26 (IH, d, J 8.8 Hz), 7.74-7.28 (13H, m), 6.76 (IH, d, J
15.6 Hz), 5.66 (IH, d, J 8.0 Hz), and 3.51 (3H, s).
Anal. Calcd. for C25H19N4O4:
C, 68.17; H, 4.58; N, 12.72. Found: C, 68.25; H, 4.65; N, 12.57%.
EXAMPLE 1 1
Figure imgf000045_0001
E-(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepm-3-yll-3-(2.4-dichlorophenvD-2-propenamide m.p. 137-139°C, [α]D +66.0° (c=0.144, CH2CI2). dH (CDCI3) 8.02 (IH, d, J 15.6 Hz), 7.73-7.26 (13H, m), 6.66 (IH, d, J
15.6 Hz), 5.81 (IH, d, J 8.8 Hz), and 3.53 (3H, s).
Anal. Calcd. for C25H19CI2N3O2:
C, 64.67; H, 4.12; N, 9.05. Found: C, 64.28; H, 4.24; N, 8.83%. EX AMPLE 12
Figure imgf000046_0001
E-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4- benzodiazepin-3-yll-3-(4-methylphenvπ-2-propenamide m.p. 133-135°C, [α]D +90.4° (c=0.125, CH2CI2). dH (CDC13) 7.68-7.19 (15H, m), 6.59 (IH, d, J 15.6 Hz), 5.70 (IH, d, J
8.0 Hz), 3.50 (3H, s), and 2.38 (3H, s).
Anal. Calcd. for C26H23N3O2:
C, 76.26; H, 5.66; N, 10.26. Found: C, 75.93; H, 5.82; N, 10.10%.
EXAMPLE 13
Figure imgf000046_0002
E-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yl1-3-(4-methoxyphenvD-2-propenamide m.p. 129-133°C, [α]D +89.9° (c 0.188, CH2CI2). dH (CDCI3) 7.65-7.24 (14H, m), 6.92 (IH, d, J 8.8 Hz), 6.50 (IH, d, J 15.6 Hz), 5.69 (IH, d, J 8.0 Hz), 3.84 (3H, s), and 3.50 (3H, s). Anal. Calcd. for C26H23N3O3.O.3OH2O:
C, 72.48; H, 5.52; N, 9.75. Found: C, 72.75; H, 5.60; N, 9.36%.
EXAMPLE 14
Figure imgf000047_0001
(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazep -3-yll-3-(2.4-dichlorophenvDpropanamide m.p. 92-95°C, [α]D 90.5° (c = 0.196, CH2CI2). dH (CDCI3) 7.62-7.15 (13H, m), 5.52 (IH, d, J 8.1 Hz), 3.47 (3H, s),
3.10 (2H, t, J 7.6 Hz), and 2.68 (2H, dd, J 7.6, 2.8 Hz).
Anal. Calcd. for C25H21CI2N3O2.O.2OH2O:
C, 63.89; H, 4.59; N, 8.94. Found: C, 63.86; H, 4.62; N, 8.87%.
EX AMPLE 15
Figure imgf000048_0001
E-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yll-3-(3-chlorophenyl)-2-propenamide m.p. 229-231 °C, [α]D +86.2° (c = 0.225, CH2CI2). dH (CDC13) 7.64-7.26 (15H, m), 6.62 (IH, d, J 15.6 Hz), 5.66 (IH, d, J
8.1 Hz), and 3.50 (3H, s).
Anal. Calcd. for C25H20CIN3O2:
C, 69.85; H, 4.69; N, 9.77. Found: C, 70.20; H, 4.83; N, 9.41%.
EXAMPLE 16
Figure imgf000048_0002
E-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yll-3-(2-chlorophenvO-2-propenamide m.p. 128-131°C, [α]D = +61.7° (c = 0.196, CH2CI2). dH (CDC13) 8.06 (IH, d, J 15.6 Hz), 7.65-7.28 (14H, m), 6.62, (IH, d, J 15.6 Hz), 5.68 (IH, d, J 8.3 Hz), and 3.50 (3H, s). Anal. Calcd. for C25H20CIN3O2.O.2OH2O:
C, 69.27; H, 4.74; N, 9.69. Found: C, 69.21 ; H, 4.68; N, 9.45%.
EXAMPLE 17
Figure imgf000049_0001
E-(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yll-3-(2.4-difluorophenyl)-2-propenamide m.p. 121-123°C, [α]D +76.8° (c = 0.1 1 1, CH2CI2). dH (CDCI3) 7.71 (IH, d, J 15.9 Hz), 7.64-7.24 (1 IH, m), 6.92-6.84 (2H, m), 6.69 (IH, d, J 15.9 Hz), 5.67 (IH, d, J 8.1 Hz), and 3.50 (3H, s). Anal. Calcd. for C25H19F2N3O2.O.IOH2O:
C, 69.31; H, 4.47; N, 9.70. Found: C, 69.28; H, 4.57; N, 9.31 %.
EX AMPLE 18
Figure imgf000050_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methyl -2-oxo-5 -phenyl- 1 H- 1 ,4-benzo- diazepin-3-yπ-3-(4-chloroρhenyDproρanamide m.p. 203-205°C, [α]D +99.2° (c = 0.300, CH2CI2). dH (CDC13) 7.62-7.16 (14H, m), 5.52 (I H, d, J 8.1 Hz), 3.47 (3H, sj,
2.99 (2H, t, J 7.7 Hz), and 2.67 (2H, t, J 7.7 Hz).
Anal. Calcd. for C25H22CIN3O2:
C, 69.52; H, 5.13; N, 9.73. Found: C, 69.50; H, 5.15; N, 9.72%.
EXAMPLE 19
Figure imgf000050_0002
E-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-pheny diazepin-3-yll-3-(2.6-dichlorophenyl)-2-propenamide m.p. 121-124°C, [α]D +69.0° (c = 0.342, CH2CI2). dH (CDCI3) 7.79 (IH, d, J 16.1 Hz), 7.64-7.15 (13H, m), 6.78 (IH, d, J 15.8 Hz), 5.69 (IH, d, J 8.1 Hz), and 3.50 (3H, s). Anal. Calcd. for C25Hl9Cl2N3θ2.0.15PhCH3:
C, 65.44; H, 4.23; N, 8.79. Found: C, 65.40; H, 4.38; N, 8.85%.
EXAMPLE 20
Figure imgf000051_0001
E-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yll-3-[4-(trifluoromethvπphenyl1-2-propenamide m.p. 133-137°C, [α]D +68.7° (c = 0.1 15, CH2CI2). dH (CDCI3) 7.72-7.25 (15H, m), 6.71 (IH, d, J 15.6 Hz), 5.67 (IH, d, J
8.1 Hz), and 3.51 (3H, s).
Anal. Calcd. for C26H20F3N3O2:
C, 67.38; H, 4.35; N, 9.07. Found: C, 67.38; H, 4.45; N, 8.95%.
EX AMPLE 21
Figure imgf000052_0001
(+)-5-Chloro-N-[(3R)-2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4- benzodiazepin-3-yllindole-2-carboxamide m.p. 160-164°C, [α]D +103.8° (c = 0.160, CH2CI2). dH (CDCI3) 9.71 (IH, br s), 8.13 (IH, d, J 7.8 Hz), 7 68-7.09 (13H, m),
5.75 (IH, d, J 7.8 Hz), and 3.53 (3H, s).
Anal. Calcd. for C25Hl9ClN4θ2.0.25H2θ.0.15PhCH3:
C, 67.84; H, 4.49; N, 12.15. Found: C, 67.80; H, 4.41; N, 12.07%.
EXAMPLE 22
Figure imgf000052_0002
(+)-N-[(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-vπ-2.2-diphenylethanamide m.p. 200-201°C, [α]D +97.0° (c = 0.168, CH2CI2). dH (CDC13) 7.60-7.22 (20H, m), 5.58 (IH, d, J 8.1 Hz), 5.08 (IH, s), and 3.44 (3H, s).
Anal. Calcd. for C3θH25N3θ2.0.15PhCH3:
C, 78.79; H, 5.55; N, 8.88. Found: C, 78.81; H, 5.63; N, 9.07%.
EXAMPLE 23
Figure imgf000053_0001
(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH- l ,4-benzo- diazepin-3-yl1-3-(2.4-difluorophenyOpropanamide m.p. 79-81°C, [α]D +92.9° (c = 0.105, CH2CI2). dH (CDCI3) 7.62-7.56 (3H, m), 7.50-7.19 (8H, m), 6.82-6.76 (2H, m),
5.52 (IH, d, J 8.1 Hz), 3.47 (3H, s), 3.01 (2H, t, J 7.6 Hz), and 2.69 (2H, m).
Anal. Calcd. for C25H21F2N3O2:
C, 69.27; H, 4.88; N, 9.69. Found: C, 68.96; H, 4.99; N, 9.47%.
EXAMPLE 24
Figure imgf000054_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-pheny 1- 1 H- 1 ,4-benzo- diazepin-3-yll-2-phenyIethanamide m.p. 241-242°C (dec), [α]D +85.5° (c = 0.159, CH2CI2). dH (CDC13) 7.59-7.55 (3H, m), 7.46-7.22 (12H, m), 5.51 (IH, d, J 8.1Hz), 3.72 (2H, s), and 3.44 (3H, s). Anal. Calcd. for C24H21N3O2.O.55H2O: C, 73.28; H, 5.66; N, 10.68. Found: C, 73.25; H, 5.38; N, 10.47%.
EXAMPLE 25
Figure imgf000054_0002
(+)-N-[(3R)-2,3-Dihydro- 1 -methyl -2-oxo-5 -phenyl- 1 H- 1 ,4-benzo- diazepin-3-vn-3-(2-chlorophenyl propanamide m.p. 158.5-159.5°C, [α]D +95.8° (c = 0.224, CH2CI2). dH (CDC13) 7.62-7.57 (3H, m), 7.47-7.16 (1 IH, m), 5.55 (IH, d, J 8.1 Hz), 3.47 (3H, s), 3.14 (2H, t, J 7.9 Hz), and 2.75-2.69 (2H, m). Anal. Calcd. for C25H22CIN3O2.O.I5H2O:
C, 69.09; H, 5.17; N, 9.67. Found: C, 69.05; H, 5.12; N, 9.63%.
EXAMPLE 26
Figure imgf000055_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yll-3-14-(trifluoromethyl)phenyllpropanamide m.p. 175-176°C, [α]D +86.5° (c = 0.141 , CH2CI2). dH (CDCI3) 7.62-7.54 (5H, m), 7.47-7.22 (9H, m), 5.52 (IH, d, J 8.1
Hz), 3.47 (3H, m), 3.08 (2H, t, J 7.6Hz), and 2.72 (2H, m).
Anal. Calcd. for C26H22F3N3O2.O.8OH2O:
C, 65.08; H, 4.93; N, 8.76. Found: C, 65.03; H, 4.63; N, 8.72%.
EXAMPLE 27
Figure imgf000056_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yl1-2-F4-(trifluoromethvπphenyllethanamide m.p. 224-226°C, [α]D +68.0° (c = 0.153, CH2CI2). dH (CDC13) 7.63-7.55 (4H, m), 7.51 -7.33 (8H, m), 7.26-7.23 (2H, m),
5.51 (IH, d, J 8.1 Hz), 3.77 (2H, s), and 3.46 (3H, s).
Anal. Calcd. for C25H20F3N3O2:
C, 66.51 ; H, 4.47; N, 9.31. Found: C, 66.46; H, 4.36; N, 9.10%.
EXAMPLE 28
Figure imgf000056_0002
(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-vn-3-r3-(trifluoromethyl)phenvnpropanamide m.p. 135-136°C, [α]D +78.8° (c = 0.134, CH2CI2). dH (CDCI3) 7.62-7.56 (3H, m), 7.49-7.22 (11H, m), 5.53 (IH, d, J 8.1 Hz), 3.47 (3H, s), 3.08 (2H, t, J 7.3 Hz), and 2.72 (2H, m). Anal. Calcd. for C26H22F3N3O2:
C, 67.09; H, 4.76; N, 9.03. Found: C, 67.03; H, 4.73; N, 9.13%.
EXAMPLE 29
Figure imgf000057_0001
(+)-3-Cyclohexyl-N-[(3R)-2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH-
1.4-benzodiazepin-3-yllpropanamide m.p. 144.5-145.5°C, [ ]D +83.1° (c = 0.1 16, CH2CI2). dH (CDCI3) 7.62-7.56 (3H, m), 7.46-7.21 (7H, m), 5.55 (IH, d, J 8.3
Hz), 3.48 (3H, s), 2.41-2.36 (2H, m), 1.77-1.58 (7H, m), 1.31-1.16 (4H, m), and 0.98-0.90 (2H, m).
Anal. Calcd. for C25H29N3O2:
C, 74.41 ; H, 7.24; N, 10.41. Found: C, 74.46; H, 7.27; N, 10.58%.
EXAMPLE 30
Figure imgf000058_0001
(+)-N-[(3R)-2,3 -Dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yll-3-12-(trifluoromethyl)phenyllpropanamide m.p. 110-113°C, [α]D +79.2° (c = 0.376, CH2CI2). dH (CDCI3) 7.65-7.57 (4H, m), 7.50-7.22 (10H, m), 5.55 (IH, d, J 8.0 Hz), 3.47 (3H, s), 3.20 (2H, t, J 7.9 Hz), and 2.70 (2H, dt, J 7.9, 3.3 Hz). Anal. Calcd. for C26H22F3N3O2:
C, 67.09; H, 4.76; N, 9.03. Found: C, 66.97; H, 4.76; N, 8.93%.
EXAMPLE 31
Figure imgf000058_0002
(+)-N-f(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yll-3-(4-cvanophenyl)propanamide m.p. 81 -85°C, [α]D +91.0° (c = 0.111 , CH2CI2). dH (CDC13) 7.64-7.55 (4H, m), 7.48-7.16 (10H, m), 5.50 (IH, d, J 8.3 Hz), 3.47 (3H, s), 3.08 (2H, t, J 7.6 Hz), and 2.74-2.69 (2H, m). Anal. Calcd. for C26H22N4θ2.0.60H2θ.0.50PhCH3:
C, 73.93; H, 5.62; N, 1 1.69. Found: C, 73.98; H, 5.61; N, 1 1.71%.
EXAMPLE 32
Figure imgf000059_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yll-3-G-chlorophenyl)propanamide m.p. 157-159°C, [α]D +90.7° (c = 0.134, CH2CI2). dH (CDCI3) 7.62-7.57 (3H, m), 7.47-7.12 (11H, m), 5.53 (IH, d, J 8.1
Hz), 3.47 (3H, s), 3.00 (2H, t, J 7.3 Hz), and 2.71-2.66 (2H, m). Anal. Calcd. for C25H22CIN3O2.O.55H2O:
C, 67.96; H, 5.27; N, 9.51. Found: C, 67.99; H, 5.18; N, 9.26%.
EXAMPLE 33
Figure imgf000060_0001
E-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yl1-3-(2-bromophenyl)-2-propenamide m.p. 113-116°C, [ ]D +44.2° (c = 0.1 13, CH2CI2). dH (CDCI3) 8.03 (IH, d, J 15.6 Hz), 7.64-7.16 (14H, m), 6.57 (IH, d, J
15.6 Hz), 5.68 (IH, d, J 8.1 Hz), and 3.50 (3H, s).
Anal. Calcd. for C25H2θBrN3θ2.0.60H2θ.0.30PhCH3:
C, 63.48; H, 4.58; N, 8.19. Found: C, 63.49; H, 4.38; N, 8.19%.
EXAMPLE 34
Figure imgf000060_0002
E-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yl1-3-G-brornophenyl)-2-propenamide m.p. 221-223 d°C, [α]D +65.5° (c = 0.206, CH2CI2). dH (CDC13) 7.69 (IH, br s), 7.64-7.57 (4H, m), 7.51-7.37 (6H, m), 7.29-7.19 (4H, m), 6.62 (IH, d, J 15.6 Hz), 5.66 (IH, d, J 8.1 Hz), and 3.50 (3H, s). Anal. Calcd. for C25H2θBrN3θ2.0.35H2θ.0.20PhCH3:
C, 63.54; H, 4.46; N, 8.42. Found: C, 63.50; H, 4.39; N, 8.42%.
EXAMPLE 35
Figure imgf000061_0001
E-(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-vπ-3-(4-iodophenvD-2-propenamide m.p. 137-140°C, [α]p +67.9° (c = 0.268, CH2CI2). dH (CDCI3) 7.75-7.72 (2H, m), 7.64-7.36 (8H, m), 7.29-7.16 (5H, m), 6.63 (IH, d, J 15.6 Hz), 5.66 (IH, d, J 8.1 Hz), and 3.50 (3H, m). Anal. Calcd. for C25H2θIN3θ2.0.30PhCH3:
C, 59.29; H, 4.06; N, 7.65. Found: C, 59.29; H, 3.90; N, 7.40%.
EXAMPLE 36
Figure imgf000062_0001
E-(-r N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yl1-3-(4-bromophenvD-2-propenamide m.p. 121-124°C, [ ]D +75.6° (c = 0.201, CH2CI2). dH (CDCI3) 7.64-7.57 (3H, m), 7.55-7.35 (1 1H, m), 7.28-7.24 (IH, m), 6.62 (IH, d, J 15.6 Hz), 5.66 (IH, d, J 8.1 Hz), and 3.50 (3H, s). Anal. Calcd. for C25H2θBrN3θ2:
C, 63.30; H, 4.25; N, 8.86. Found: C, 63.50; H, 4.20; N, 8.78%.
EXAMPLE 37
Figure imgf000062_0002
(+)-N-[(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yl1-4-phenylbutanamide m.p. 65-74°C, [α]D +77.4° (c = 0.155, CH2CI2). dH (CDCI3) 7.62-7.56 (3H, m), 7.46-7.19 (12H, m), 5.55 (IH, d, J 8.1 Hz), 3.47 (3H, s), 2.71 (2H, t, J 7.6 Hz), 2.42-2.37 (2H, m), and 2.09- 2.01 (2H, m). Anal. Calcd. for C26H25N3O2.OJOH2O: C, 74.91 ; H, 6.19; N, 10.08. Found: C, 74.93; H, 6.05; N, 10.07%.
EXAMPLE 38
Figure imgf000063_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-pheny 1- 1 H- 1 ,4-benzo- diazepin-3-yll-5-methyl-3-phenylisoxazole-4-carboxamide m.p. 123-126°C, [α]D +122.0° (c = 0.199, CH2CI2). dH (CDC13) 7.79-7.76 (2H, m), 7.62-7.32 (11H, m), 7.26-7.21 (2H, m), 5.61 (IH, d, J 7.9 Hz), 3.42 (3H, s), and 2.76 (3H, s). Anal. Calcd. for C27H22N4O3.O.4OH2O: C, 70.85; H, 5.02; N, 12.24. Found: C, 70.84; H, 4.91; N, 11.92%.
EXAMPLE 39
Figure imgf000063_0002
(+)-N-[(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yll-3-(3-cyanophenyl)ρropanamide m.p. 1 10-112°C, [α]D +84.2° (c = 0.202, CH2CI2). dH (CDCI3) 7.63-7.22 (14H, m), 5.51 (IH, d, J 8.1 Hz), 3.47 (3H, s), 3.06 (2H, t, J 7.8 Hz), and 2.74-2.68 (2H, m). Anal. Calcd. for C26H22N4O2.0.50H2O: C, 72.37; H, 5.37; N, 12.98. Found: C, 72.52; H, 5.12; N, 12.59%.
EXAMPLE 40
Figure imgf000064_0001
(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yllcvclohexanethanamide m.p. 144-146°C, [α]D +72.1° (c=1.000, MeOH). Anal. Calcd. for C24H27N3O2.O.2OH2O: C, 73.33; H, 7.03; N, 10.69. Found: C, 73.27; H, 7.02; N, 10.76%. EX AMPLE 41
Figure imgf000065_0001
(+)-4-Cyclohexyl-N-[(3R)-2,3-dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H-
1.4-benzodiazepin-3-yllbutanamide
[α]D +57.7° (c=0.440, MeOH). Anal. Calcd. for C26H31N3O2:
C, 74.79; H, 7.48; N, 10.06. Found: C, 74.8;0 H, 7.78; N, 10.05%.
EXAMPLE 42
Figure imgf000065_0002
(+)-N-[(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yn-4-methylpentanamide m.p. 123-125°C, [α]D +66.8° (c=0.500, MeOH). Anal. Calcd. for C22H25N3O2.0.45H2O: C, 71.12; H, 7.03; N, 11.31. Found: C, 71.08; H, 6.81; N, 1 1.42%. EXAMPLE 43
Figure imgf000066_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yll-2.3-dihydrobenzofuran-2-carboxamide
Diisopropylethylamine (0.3 mL, 223 mg, 1.72 mmol) was added to a stirred, cooled (0 °C) solution of 3(R)-amino-l ,3-dihydro-l - methyl-5-phenyl-2H-l ,4-benzodiazepin-2-one (J. Org. Chem. 1987, 52, 3232-3239) (400 mg, 1.5 mmol), 2,3-dihydrobenzofuran-2-carboxylic acid (274 mg, 1.7 mmol), l-(3-dimethylaminopropyl)-3-ethylcarbodi- imide hydrochloride (583 mg, 3.0 mmol), and 1-hydroxybenzotriazole (479 mg, 3.1 mmol) in DMF (4.5 mL). The mixture was stirred at room temperature for 18 h., poured into aqueous hydrochloric acid (3M, 12 mL) and extracted with ethyl acetate (3 x 20 mL). The combined organic fractions were washed with saturated aqueous sodium hydrogen carbonate (20 mL) and brine (20 mL), dried (MgSθ4) and evaporated under reduced pressure. The residue was crystallized from 2-chloro-2-methylpropane/hexane to give (+)-N-[(3R)-2,3-dihydro-l- methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzodiazepin-3-yl]-2,3-dihydrobenzo- furan-2-carboxamide as a colorless solid (156 mg, 25%), m.p. 141- 180°C, [α]D +127.1° (c=0.425, CHCI3). dH (CDCI3) (3: 1 Mixture of diastereoisomers) 8.44 (IH, m), 7.65-6.91 (13H, m), 5.52 (IH, m), 5.28 (IH, m), and 3.70-3.40 (5H, m). Anal. Calcd. for C25H21N3O3.O.25 Hexane
C, 73.50; H, 5.70; N, 9.71. Found: C, 74.12; H, 5.57; N, 9.71%. EXAMPLE 44
(+)-N-[(3R)-2, 3-dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yl]-l'-(l,l -dimethylethoxycarbonyl)spiro(cyclohexan-4,4'- piperidine 1 -carboxamide
Step A:
Figure imgf000067_0001
Diethyl 1 -benzylpiperidine-4.4-diacetate
Ethanol (120 mL) was cooled in ice and ammonia bubbled through to give a saturated solution. 1 -Benzyl-4-piperidone (40.0g, 21 lmmol) and ethyl cyanoacetate (47.8g, 423 mmol) were added, the reaction vessel stoppered and stored at 0°C overnight. The solid was collected, washed with ethanol and ether and dried in vacuo to give a yellow solid (68.86g). The solid (58.86g) was dissolved in a mixture of sulfuric acid (70 mL, 98%) and water (60 mL) and heated under reflux for three days the mixture cooled and most of the water evaporated. The residue was azeotroped with ethanol (4x750 mL), further ethanol (500 mL) added and the mixture heated under reflux for 20h, cooled in ice and sodium carbonate (lOOg) added slowly with vigorous stirring. The ethanol was evaporated under reduced pressure, water (800 mL) added and the mixture extracted with methylene chloride (3x400 mL). The combined organic extracts were dried (Na2Sθ4) and the solvent evaporated to give diethyl l-benzylpiperidine-4,4-diacetate (37.5 lg). A small portion of this was purified by flash column chromatography. NMR (300 MHz, CDCI3) d: 7.2-7.4 (m, 5H), 4.1 1 (q, J=7.3Hz,4H), 3.50 (s, 2H), 2.56 (s, 4H), 2.4 (m, 4H), 1.7 (m, 4H), 1.24 (t, J=7.3Hz, 6H).
Step B:
Figure imgf000068_0001
1 -Benzylpiperidine-4.4-diethanol
A solution of the diester (12.2 g, 35 mmol) in ether (25 mL) was added to a cooled (-30°C) and stirred suspension of LiAlH4 (2.1 g, 55 mmol) in ether (400 mL), under argon. THF (60 mL) was added and the reaction mixture allowed to warm to room temperature. After recooling to 0°C, water (2.2 mL), IM NaOH (4.4 mL) and water (5 mL) were added, the reaction mixture stirred vigorously for 30 min and the solid filtered off, washing well with ether. The combined filtrates were evaporated to afford a white solid which was tritutrated with ether to give 8 g of l-benzylpiperidine-4,4-diethanol. m.p. 75-78°C
NMR (300 MHz, CDCI3) d: 7.2-7.4 (m, 5H), 3.7 (t, J = 6.8 Hz, 4H), 3.52 (s, 2H), 2.7 (brs, 2H), 2.43 (m, 4H), 1.66 (t, J = 6.8 Hz, 4H), 1.5 (m, 4H).
Step C:
Figure imgf000068_0002
l-t-Butoxycarbonylpiperidine-4.4-diethanol
The benzylamine (2.07 g, 7.9 mmol) was dissolved in methanol (60 mL), BOC2O (1.72 g, 7.9 mmol) added and the mixture hydrogenated at 50 psi over 10% palladium hydroxide on charcoal (200 mg) for 18 hours. The reaction mixture was filtered through celite, washed with methanol and the filtrate evaporated to give 1 -t-butoxy- carbonylpiperidine-4,4-diethanol (2.0 g).
NMR (300 MHz, CDCI3) d: 3.7 (m, 4H), d 3.3 (m, 6H), 1.65 (t, J = 6.8 Hz, 4H), 1.41 (s, 9H).
Step D:
Figure imgf000069_0001
1 -t-Butoxycarbonylpiperidine-4.4-diethanol. bis(methanesulfonate The diol (2.41 g, 8.9 mmol) was dissolved in dichloro- methene (50 mL), the solution cooled to -20°C under argon before addition of triethylamine (3.7 mL, 26 mmol) and methanesulfonyl chloride (1.6 mL, 20 mmol). After 30 min., the reaction mixture was poured into ice cold 10% citric acid and extracted with ether (X3). The combined extracts were washed with water, saturated NaHCθ3 and brine, dried (MgS04) and the solvent evaporated to afford 1-t-butoxy- carbonylpiperidine-4,4-diethanol, bis(methanesulfonate) (3.2g). NMR (300 MHz, CDCI3) d: 4.32 (t, J = 7.1 Hz, 4H), 3.4 (m, 4H), 3.04 (s, 6H), 1.89 (t, J = 7.1 Hz, 4H). Step E:
Figure imgf000070_0001
Diethyl 3-t-butyloxycarbonyl-3-azaspiro[5.51undecane-9.9-dicarboxylate
To a slurry of 60% NaH (2.04 g, 0.51 mole) in toluene (160 mL), under argon, was slowly added diethyl malonate (3.72 mL, 24.3 mmol). The mixture was cooled to 0°C and the bis-mesylate 1 (7.0 g, 16.3 mmol) added as a solid and the mixture heated to reflux for 18 hours. The reaction was quenched into 10% citric acid (100 mL) and the product extracted with CH2CI2 (2x150 mL). The extracts were dried (Na2Sθ4), concentrated to an oil, and chromatographed on silica to give 3.83 g (60% ) of diethyl 3-t-butyloxycarbonyl-3-azaspiro- [5.5]undecane-9,9-dicarboxylate.
IH NMR (CDCI3) d: 1.22 (t, 6H), 1.4 (s, 9H), 2.0 (m, 4H), 3.35 (m, 4H), 4.2 (q, 4H).
Step F: H
Figure imgf000070_0002
3-t-Butyloxycarbonyl-3-azaspirol5.51undecane-9-carboxylic acid
To a solution of the diester 2 (3.69 g, 0.0093 m) in THF (50 mL) was added IN LiOH (47 mL). The reaction was stirred for 3 days at 25°C, diluted with water (50 mL) and pH adjusted to 2.2 with KHSO4. The product was extracted into ethyl acetate (2x75 mL), dried (Na2Sθ4), and concentrated to a foam (3.5 g). The solid was melted in a flask at 140°C for 2 hours, cooled and the oil dissolved in THF (15 mL), IN LiOH (10 mL) added and mixture stirred overnight at 30°C. The reaction was concentrated to remove THF, diluted with water (20 mL) and washed with diethyl ether (10 mL). The pH was adjusted to 2.5 with KHSO4 and product extracted (3x50 mL) with ethyl acetate. The extracts were dried (Na2Sθ4), filtered and concentrated to yield 3- t-butyloxycarbonyl-3-azaspiro[5.5]undecane-9-carboxylic acid as a foam (2.48 g, 90%). I H NMR (CDCI3, partial) d: 1.45 (s, 9H), 3.4 (m, 4H).
Employing the procedure substantially as described in Example 43 but substituting an appropriate acid for the 2,3-dihydro- benzofuran-2-carboxylic acid, the following compounds were prepared:
Step G:
Figure imgf000071_0001
(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l,4-benzo- diazepin-3-yl]-l '-(l ,1 -dimethylethoxycarbonyl)spiro(cyclohexan-4,4'- piperidineV 1 -carboxamide m.p. 135-138°C, [α]D +58.8° (C=0.925, CHCI3). dH (CDCI3) 7.61-7.23 (10H, m), 5.54 (IH, d, J 9.0 Hz), 3.47 (3H, s), 3.37 (4H, m), 2.28 (IH, m), and 1.81 -1.18 (21H, s). Anal. Calcd. for C32H40N4O4:
C, 70.56; H, 7.40; N, 10.29. Found: C, 70.21; H, 7.40; N, 10.16%.
EXAMPLE 45
Figure imgf000072_0001
(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH- diazepin-3-yll-3-(furan-2-yDpropanamide m.p. 115-1 18°C, [α]D +65.8° (c=0.800, CHCI3). dH (CDCI3) 7.62-7.26 (11H, m), 6.28 (IH, dd, J 3.2, 2.0 Hz), 6.08 (IH, dd, J 3.2, 0.7 Hz), 5.58 (IH, d, J 8.1 Hz), 3.48 (3H, s), 3.04 (2H, t, J 7.6
Hz), and 2.75 (2H, m).
Anal. Calcd. for C23H2lN3θ3.0.3Hexane:
C, 72.07; H, 6.15; N, 10.17. Found: C, 71.78; H, 6.30; N, 9.77%.
EXAMPLE 46
Figure imgf000073_0001
(+)-N- [(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-v11-4-(2-thienvπbutanamide m.p. 170-180°C, [α]D +63.5° (c=1.000, MeOH). Anal. Calcd. for C24H23N3O2S.O.95H2O:
C, 66.32; H, 5.77; N, 9.67. Found: C, 66.32; H, 5.34; N, 9.40%.
EXAMPLE 47
Figure imgf000073_0002
(+)-N-[(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yllcvclohexylcarboxamide m.p. 213-214°C, [ ]D +62.4° (c=1.000, MeOH). Anal. Calcd. for C23H24N3O2:
C, 73.77; H, 6.46; N, 1 1.22. Found: C, 73.86; H, 6.81; N, 1 1.15%. EXAMPLE 48
Figure imgf000074_0001
(E)-(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l,4-benzo- diazepin-3-yH-3-(3.4-methylenedioxyphenvD-2-propenamide m.p. 143-145°C, [α]D +62.3° (c=0.960, MeOH). Anal. Calcd. for 25H2lN3θ4.0.10H2θ.0.20Et2θ:
C, 69.78; H, 5.27; N, 9.46. Found: C, 69.78; H, 4.98; N, 9.28%.
EXAMPLE 49
Figure imgf000074_0002
(+)-N-[(3R)-2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yll-2-quinoxalinecarboxamide
[α]D +85.8° (c=0.360, MeOH). Anal. Calcd. for C25H19N5O2:
C, 69.96; H, 4.90; N, 15.33. Found: C, 69.95; H, 4.72; N, 15.25%.
EXAMPLE 50
(+)-N-[(3R)-2,3-Dihydro-2-methyl-2-oxo-5-phenyl-lH-l,4-benzo- diazepin-3-yl1-2-(phenylamino)acetamide
Step A:
Figure imgf000075_0001
N-[(3R)-2,3-Dmydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzodiazepin-
3-vn-2-bromoacetamide
Bromoacetyl bromide (165 mL, 383 mg, 1.9 mmol) was added to an ice cooled solution of 3(R)-amino-l ,3-dihydro-l-methyl-5- phenyl-2H-l ,4-benzodiazepin-2-one (J. Org. Chem. 1987, 52, 3232- 3239) (500 mg, 1.88 mmol) and triethylamine (264 mL, 192 mg, 1.9 mmol) in methylene chloride (10 mL) and the mixture was stirred at room temperature for 1 h. The mixture was washed with water (3 x 10 mL), dried (MgSθ4) and the solvent was evaporated under reduced pressure to give N-[(3R)-2,3-dihydro-l -methyl -2-oxo-5-phenyl-lH- 1 ,4- benzodiazepin-3-yl]-2-bromoacetamide as a colorless foam (760 mg,
100%). dH (CDC13) 8.24 (IH, d, J 7.8 Hz), 7.64-7.24 (9H, m), 5.48 (IH, d, J
7.8 Hz), 4.00 (2H, m), and 3.50 (3H, s). Step B:
Figure imgf000076_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yl1-2-(phenylamino acetamide
Aniline (297 mL, 304 mg, 3.26 mmol) was added to a solution of N-[(3R)-2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4- benzodiazepin-3-yl]-2-bromoacetamide (600 mg, 1.55 mmol) in ethanol (25 mL) and the mixture was heated under reflux for 24 h. The mixture was cooled and the solid was collected and recrystallized from ethanol (20 mL) to give (+)-N-[(3R)-2,3-dihydro-l-methyl-2-oxo-5- phenyl-lH-l,4-benzodiazepin-3-yl]-2-(phenylamino)acetamide as a colorless solid (500 mg, 81 %), m.p. 245-246°C, [α]D +119° (C=0.850, CHC13). dH (CDC13) 8.26 (IH, d, J 8.3 Hz), 7.63-7.20 (12H, m), 6.81 (IH, t, J 7.3 Hz), 6.72 (2H, d, J 7.6 Hz), 5.56 (IH, d, J 8.3 Hz), 3.95 (2H, d, J 1.5 Hz), and 3.45 (3H, s). Anal. Calcd. for C24H22N4O2:
C, 72.34; H, 5.57; N, 14.06. Found: C, 72.37; H, 5.59; N, 14.32%.
Employing the procedure substantially as described above, but substituting 2-chloroaniline or 4-(trifluoromethyl aniline for the aniline, the following compounds were prepared: EX AMPLE 51
Figure imgf000077_0001
(- -N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yll-2-(2-chlorophenylamino acetamide m.p. 222-224°C, fα]D +1 11° (c=0.973, CHCI3). dH (CDCI3) 8.15 (IH, d, J 8.3 Hz), 7.60-7.16 (12H, m), 6.71 (2H, m), 5.57 (IH, d, J 8.3 Hz), 4.01 (2H, d, J 2.7 Hz), and 3.45 (3H, s). Anal. Calcd. for C24H21CIN4O2:
C, 66.59; H, 4.89; N, 12.94. Found: C, 66.40; H, 4.94; N, 12.92%.
EXAMPLE 52
Figure imgf000077_0002
(+)-N-[(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yπ-2-r4-(trifluoromethvDphenylaminolacetamide m.p. 218-219°C, [αlD +91.9° (c = 0.419, CHCI3). dH (CDC13) 8.13 (IH, d, J 9.0 Hz), 7.70-7.25 (12H, m), 6.72 (2H, d, J 8.7 Hz), 5.60 (IH, d, J 9.0 Hz), 4.05 (2H, m), and 3.50 (3H, s). Anal. Calcd. for C25H21F3N4O2.O.7H2O:
C, 62.68; H, 4.71 ; N, 11.69. Found: C, 62.47; H, 4.32; N, 11.44%.
EXAMPLE 53
Figure imgf000078_0001
(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yl1-2-(phenoxy)acetamide
Phenol (104 mg, 1.1 mmol) was added to a suspension of sodium hydride (60% dispersion in mineral oil, 44 mg, 1.1 mmol) in toluene (10 mL). When hydrogen evolution had stopped, N-[(3R)-2,3- dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4-benzodiazepin-3-yl] -2-bromo- acetamide (400 mg, 1.04 mmol) was added and the mixture was stirred at room temperature for 1 h. The mixture was washed with water (3 x 15 mL), dried (MgS04) and the solvent was evaporated under reduced pressure. The residue was triturated with 2-propanol and the solid was collected and recrystallized from 2-propanol (5 mL) to give (+)-N- [(3R)-2,3-dihydro-l -methyl-2-oxo-5-phenyl-lH-l,4-benzodiazepin-3- yl]-2-(phenoxy)acetamide as a colorless solid (112 mg, 27%), m.p. 126- 128°C, [α]D +81.6 (C=0.692, CHCI3). dH (CDCI3) 8.49 (IH, d, J 8.2 Hz), 7.64-7.01 (14H, m), 5.61 (IH, d, J 8.2 Hz), 4.65 (IH, d, J 14.6 Hz), 4.58 (IH, d, J 14.6 Hz), and 3.50 (3H, s). Anal. Calcd. for C24H21 N3O3:
C, 72.17; H, 5.30; N, 10.52. Found: C, 71.84; H, 5.25; N, 10.41 %.
Employing the procedure substantially as described above, but substituting 2,4-dichlorophenol, thiophenol or 2,4-dichloro- thiophenol for the phenol, the following compounds were prepared:
EXAMPLE 54
Figure imgf000079_0001
(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l,4-benzo- diazepin-3-yll-2-(2.4-dichlorophenoxy acetamide m.p. 206°C, [α]D +31.1° (c=0.289, CHCI3). dH (CDCI3) 8.75 (IH, d, J 9.0 Hz), 7.65-7.20 (1 IH, m), 6.90 (IH, d, J 8.7 Hz), 5.60 (IH, d, J 9.0 Hz), 4.65 (2H, m), and 3.50 (3H, s). Anal. Calcd. for C24H19CI2N3O3.O.3H2O:
C, 60.85; H, 4.17; N, 8.87. Found: C, 60.80; H, 4.04; N, 8.87%. EXAMPLE 55
Figure imgf000080_0001
(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzo- diazepin-3-yl1-2- henylthio)acetamide
[α]D +104.9° (c=0.316, CHC13). dH (CDCI3) 8.50 (IH, d, J 9.0 Hz), 7.60-7.20 (14H, m), 5.50 (IH, d, J
9.0 Hz), 3.75 (2H, m), and 3.45 (3H, s).
Anal. Calcd. for C24H21N3O2S:
C, 69.37; H, 5.10; N, 10.1 1. Found: C, 68.98; H, 5.06; N, 9.76%.
EXAMPLE 56
Figure imgf000080_0002
(+)-N-[(3R)-2,3-Dihydro-l -methyl-2-oxo-5-phe diazepin-3-yH-2-(2.4-dichlorophenylthio acetamide
[α]D +97.4° (c=0.286, CHCI3). dH (CDC13) 8.35 (IH, d, J 9.0 Hz), 7.70-7.20 (12H, m), 5.50 (IH, d, J 9.0 Hz), 3.70 (2H, m), and 3.50 (3H, s). Anal. Calcd. for C24H19CI2N3O2S:
C, 59.51 ; H, 3.95; N, 8.67. Found: C, 59.32; H, 3.95; N, 8.65%.
EXAMPLE 57
Figure imgf000081_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methy l-2-oxo-5-phenyl- IH- 1 ,4-benzo- diazepin-3-yll-3-(phenylamino)propanamide
3-Bromopropionyl chloride (2.01 mL, 3.428 g, 20 mmol) was added to an ice cooled solution of 3(R)-amino-l ,3-dihydro-l - methyl-5-phenyl-2H-l ,4-benzodiazepin-2-one (J. Orb. Chem. 1987, 52, 3232-3239) (5.0 g, 18.8 mmol) and triethylamine (2.79 mL, 2.02 mg, 20 mmol) in methylene chloride (85mL) and the mixture was stirred at room temperature for 18 h. The mixture was washed with saturated aqueous sodium hydrogen carbonate (85 mL), water (2 x 85 mL), and brine (85 mL), dried (MgSθ4) and the solvent was evaporated under reduced pressure. A sample (0.5 g, 1.25 mmol) was dissolved in ethanol (25 mL), aniline (230 mL, 233 mg, 2.5 mmol) was added and the mixture was heated under reflux for 70 h. The mixture was cooled and the solid was collected and recrystallized from ethanol to give (+)- N-[(3R)-2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4-benzodiazepin-3- yl]-3-(phenylamino)propanamide as a colorless solid, m.p. 218-221 °C, [α]D +58.2° (c=0.585, CHCI3). dH (CDCI3) 7.60-6.71 (16H, m), 5.54 (IH, d, J 8.1 Hz), 3.54 (2H, t, J 6.1 Hz), 3.52 (3H, s), and 2.70 (2H, m). Anal. Calcd. for C25H24N4θ2.0.5Et0H: C, 71.70; H, 6.25; N, 12.87. Found: C, 71.42; H, 5.98; N, 12.84%.
EXAMPLE 58
Figure imgf000082_0001
(+)- 1 -[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4-benzo- diazepin-3-yll-3-(2.4-dichlorophenv urea
2,4-Dichlorophenylisocyanate (188 mg, 1.0 mmol) was added to a solution of 3(R)-amino- 1 ,3 -dihydro- 1 -methy 1-5 -phenyl -2H- l ,4-benzodiazeρin-2-one (J. Org. Chem. 1987, 52, 3232-3239) (265 mg, 1.0 mmol) in tetrahydrofuran (20 mL). The mixture was stirred at room temperature for 18 h. and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2θ2/MeOH (99.5:0.5) and the residue was crystallized from CH2θ2/hexane to give (+)-l -
[(3R)-2,3-dihydro-l -methy l-2-oxo-5-phenyl-lH-l ,4-benzodiazepin-3- yl]-3-(2,4-dichlorophenyl)urea as a colorless solid, m.p. 215-216.5°C, [α]D +76.2° (c=0.261, CHCI3). dH (CDCI3) 8.10 (IH, d, J 9.0 Hz), 7.65-6.95 (13H, m), 5.50 (IH, d, J 9.0 Hz), and 3.50 (3H, s). Anal. Calcd. for C23HI8CI2N4O2.O.3H2O: C, 60.22; H, 4.09; N, 12.21. Found: C, 60.28; H, 3.89; N, 12.10%.
EXAMPLE 59
Figure imgf000083_0001
(-)-3-Cyclohexyl-N-[(3R)-2,3-dihydro-l-methyl-2-oxo-4-oxido-5- phenyl-lH- 1.4-benzodiazepin-3-yllpropanamide
3-Chloroperoxybenzoic acid (80%, 0.32 g, 1.5 mmol) was added to a solution of (+)-3-cyclohexyl-N-[(3R)-2,3-dihydro-l-methyl- 2-oxo-5-phenyl-lH-l,4-benzodiazepin-3-yl]propanamide (0.60 g, 1.5 mmol) in dichloromethane (25 mL) and the mixture was stirred at room temperature for 18 h. Further 3-chloroperoxybenzoic acid (80%, 0.1 g, 0.5 mmol) was added and the mixture was stirred for 24 h. The mixture was washed with saturated aqueous sodium hydrogen carbonate (4 x 25 mL), water (2 x 25 mL) and brine (25 mL), dried (MgSθ4) and the solvent was evaporated under reduced pressure. The residue was recrystallized from toluene/hexane (65:35) to give (-)-3-cyclohexyl-N- [(3R)-2,3-dihydro- 1 -methy l-2-oxo-4-oxido-5-pheny 1- 1 H- 1 ,4- benzodiazepin-3-yl]propanamide as colorless prisms, m.p. 222-224°C, [α]D -80.7° (c=1.15, CHCl3). dH (CDC13) 7.71-7.23 (10H, m), 6.01 (IH, d, J 9.3 Hz), 3.54 (3H, s), 2.48 (2H, m), and 1.76-0.89 (13H, m). Anal. Calcd. for C25H29N3O3.O.5H2O:
C, 70.06; H, 7.06; N, 9.81. Found: C, 70.10; H, 6.80; N, 9.79%. EXAMPLE 60
N-[2,3-Dihydro- 1 -(2-dimethy laminoethyl)-2-oxo-5-phenyl- 1 H- 1 ,4- benzodiazepin-3-yll-3-(2.4-dichlorophenyl propanamide
Step A:
Figure imgf000084_0001
2,3-Dihydro- 1 -(2-dimethylaminoethyl)-5-phenyl- 1 H- 1 ,4-benzodiazepin-
2-one
2,3-Dihydro-5 -phenyl- 1 H- 1 ,4-benzodiazepin-2-one (1.00 g, 4.23 mmol) was added to hexane washed sodium hydride (60% dispersion in mineral oil, 186 mg, 4.65 mmol) in DMF (5 mL). Further DMF (10 mL) was added and the mixture was stirred at room temperature. 2-(Dimethylamino)ethyl chloride hydrochloride (0.73 g, 5 mmol) was added to hexane washed sodium hydride (60% dispersion in mineral oil, 200 mg, 5.0 mmol) in DMF (5 mL) and the mixtures were combined. Potassium iodide (1 crystal) was added and the mixture was stirred at 110°C for 30 min. The solvent was evaporated under reduced pressure, water was added and the mixture was extracted with ethyl acetate. The combined organic fractions were washed with water (2 x), dried (MgSθ4) and the solvent was evaporated under reduced pressure to give 2,3-dihydro-l-(2-dimethylaminoethyl)-5-phenyl-lH-l,4- benzodiazepin-2-one (1.21 g, 93%). dH (CDC13) 7.63-7.16 (9H, m), 4.77 (IH, d, J 10.6 Hz), 4.41 (IH, m), 3.80 (IH, m), 3.78 (IH, d, J 10.6 Hz), 2.49 (2H, m), and 2.13 (6H, s).
Step B:
Figure imgf000085_0001
2,3-Dmydro-l-(2-dimethylaminoethyl)-3-hydroxyimino-5-phenyl-lH-
1 ,4-benzodiazepin-2-one
2,3-Dihydro- 1 -(2-dimethy laminoethyl)-5-pheny 1- 1 H- 1 ,4- benzodiazepin-2-one (1.21 g, 3.9 mmol) was dissolved in toluene (20 mL). The mixture was cooled to -78 °C and potassium t-butoxide (1.0M solution in f-butanol, 4.72 mL, 4.72 mmol) was added. The mixture was stirred at -78 °C for 20 min., then isoamyl nitrite (0.63 mL, 0.55 g, 4.72 mmol) was added. The mixture was stirred at -78°C for 90 min. then allowed to warm to room temperature and poured into aqueous citric acid (IM, 10 mL). The pH was adjusted to 5.0 with aqueous sodium hydroxide then to 7.0 with saturated aqueous sodium hydrogen carbonate. The mixture was extracted with ethyl acetate (50 mL) and the organic layer was aged at room temperature. The solid which formed was collected and dried in vacuo to give 2,3-dihydro-l- (2-dimethylaminoethyl)-3-hydroxyimino-5-phenyl-lH-l,4- benzodiazepin-2-one (0.876 g, 66%) as a solid, m.p. 232-234°C. dH (d6-DMSO) 10.90 (IH, s), 7.72-7.25 (9H, m), 4.40 (IH, m), 3.80 (IH, m), 2.50 (2H, m), and 1.85 (6H, s). Step C:
Figure imgf000086_0001
3-Amino-2,3 -dihydro- 1 -(2-dimethy laminoethyl)-5-pheny 1- 1 H- 1 ,4- benzodiazepin-2-one
Ethyl isocyanate (320 mL, 287 mg, 4.0 πunol) was added to a mixture of 2,3-dihydro-l -(2-dimethy laminoethy l)-3-hydroxyimino-5- phenyl-lH-l ,4-benzodiazepin-2-one (0.91 g, 2.7 mmol) and triethylamine (0.56 mL, 0.41 g, 4.0 mmol) in THF (30 mL). The mixture was heated under reflux for 7 h., further ethyl isocyanate (167 mL, 150 mg, 2.1 mmol) was added and the mixture was heated under reflux for 12 h. The mixture was cooled, the solvent was evaporated under reduced pressure and ethyl acetate (75 mL) and water (25 mL) were added. The organic phase was washed with water (4 x 25 mL), dried (MgS04) and evaporated under reduced pressure. The residue was dissolved in ethanol (100 mL), palladium on carbon (10%, 100 mg) was added and the mixture was shaken under hydrogen (50 p.s.i.) for 4.5 h. Further palladium on carbon (10%, 100 mg) was added and the mixture was shaken under hydrogen (50 p.s.i.) for 1.5 h. The mixture was filtered and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2Cl2/MeOH to give 3-amino-2,3-dihydro-l -(2-dimethy laminoethy I)- 5-phenyl-lH-l ,4-benzodiazepin-2-one (180 mg, 17%). dH (CDC13) 7.75-7.17 (9H, m), 4.45 (IH, s), 4.40 (IH, m), 3.82 (IH, m), 2.47 (4H, m), and 2.08 (6H, s). Step E:
Figure imgf000087_0001
N-[2,3-Dihydro- 1 -(2-dimethy laminoethy l)-2-oxo-5-pheny 1- 1 H- 1 ,4- benzodiazepin-3-yl1-3-(2.4-dichlorophenyl)propanamide
Triethylamine was added to a mixture of 3-amino-2,3- dihydro- 1 -(2-dimethylaminoethyl)-5-phenyl- 1 H- 1 ,4-benzodiazepin-2- one (180 mg, 0.6 mmol), 3-(2,4-dichlorophenyl)propanoic acid (131 mg, 0.6 mmol), l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (115 mg, 0.6 mmol) and 1 -hydroxyberzotriazole (81 mg, 0.6 mmol) in DMF (15 mL) until the pH was 9.0. The mixture was stirred at room temperature for 72 h. The solvent was evaporated under reduced pressure and ethyl acetate was added. The mixture was was washed with water, saturated aqueous sodium hydrogen carbonate and water, dried (MgS04) and evaporated under reduced pressure. The residue was triturated with acetone and recrystallized from i- PrOH/MeOH to give N-[2,3-dihydro-l-(2-dimethylaminoethyl)-2-oxo-5- phenyl-lH-l,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)-propanamide as a solid, m.p. 199-201 °C. dH (CDCI3) 7.60-7.15 (13H, m), 5.50 (IH, d, J 8.0 Hz), 4.40 (IH, m), 3.80 (IH, m), 3.10 (2H, t, J 7.5 Hz), 2.70 (2H, t, J 7.5 Hz), 2.40 (2H, m), and 2.05 (6H, s). Anal. Calcd. for C28H28CI2N4O2:
C, 64.25; H, 5.39; N, 10.70. Found: C, 64.23; H, 5.40; N, 10.61%. EX AMPLE 61
Figure imgf000088_0001
(+)-3(R)- { N-[3-(4-chlorophenyl)prop- 1 -en-3-yl]amino } - 1 ,3-dihydro-
1 -methyl-5-phenyl-2H- 1.4-benzodiazepin-2-one hydrochloride
A mixture of 3(R)-amino-l,3-dihydro-l-methyl-5-phenyl- 2H-l,4-benzodiazepin-2-one (J. Org. Chem. 1987, 52, 3232-3239) (265 mg, 1 mmol), £-l-chloro-4-(3-chloro-l-propenyl)benzene (281 mg, 1.5 mmol), potassium carbonate (276 mg, 2 mmol) and potassium iodide (25 mg, 0.15 mmol) in acetonitrile (2 mL) was heated under reflux for 4 h. The mixture was cooled and poured into ethyl acetate (10 mL) and water (5 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (5 mL). The combined organic fractions were washed with brine, dried (Na2Sθ4) and the solvent was evaporated under reduced pressure. The residue was purified by flash column chroma-tography on silica gel, eluting with EtOAc/Hexane (65:35 increasing to 100:0). The first compound to elute was suspended in ethanol (1 mL) and ethanolic HC1 (6 M, 0.11 mL) was added. The mixture was stirred, then the solvent was evaporated under reduced pressure. The residue was triturated with ether and the solid was collected and dried in vacuo to give (+)-3(R)-{N,N-bis[l-(4-chloro- phenyl)propen-3-yl]amino } - 1 ,3-dihydro-l -methyl-5-phenyl-2H- 1 ,4- benzodiazepin-2-one hydrochloride (235 mg, 39%) as a tan solid, m.p. 138-145°C, [α]D +9.2° (c=0.500, MeOH). dH (dό-DMSO) 11.2 (IH, br s), 7.77-7.31 (17H, m), 6.85 (2H, br m), 6.54 (2H, m), 5.20 (IH, br s), 4.60-4.00 (4H, m), and 3.46 (3H, s). Anal. Calcd. for C34H29Cl2N3θ.HC1.0.10EtOH:
C, 67.60; H, 5.08; N, 6.92. Found: C, 67.60; H, 5.03; N, 7.03%.
The second compound to elute was suspended in ethanol (0.5 mL) and ethanolic HC1 (6 M, 0.035 mL) was added. The mixture was stirred, then the solvent was evaporated under reduced pressure. The residue was triturated with ether and the solid was collected and dried in vacuo to give (+)-3(R)-{N-[3-(4-chlorophenyI)propen-3- yljamino } -1 ,3-dihydro- 1 -methyl-5-pheny 1-2H- 1 ,4-benzodiazepin-2-one hydro-chloride (56 mg, 12%) as a yellow solid, m.p. 156-162°C, [α]D +35° (c=0.100, MeOH). dH (d6-DMSO) 10.3 (IH, br s), 10.0 (IH, br s), 7.79-7.34 (13H, m), 6.78 (IH, d, J 15.9 Hz), 6.40 (IH, dt, Jd 15.9, Jt 9.0 Hz), 5.13 (IH, s), 4.00 (2H, m), and 3.46 (3H, s). Anal. Calcd. for C25H22ClN3θ.HC1.0.10EtOH.0.40H2θ:
C, 65.20; H, 5.30; N, 9.05. Found: C, 65.14; H, 5.09; N, 9.33%.
Employing the procedure substantially as described above, but substituting l-(2-bromoethoxy)-4-nitrobenzene or 4-chlorobenzene - propanol methanesulfonate for the E-l-chloro-4-(3-chloro-l-propenyl)- benzene, the following compounds were prepared:
EXAMPLE 62
Figure imgf000090_0001
(+)-3(S)-{N,N-Bis[2-(4-nitrophenoxy)ethyl]amino}-l,3-dihydro- 1 -methyl-5-phenyl-2H- 1 ,4-benzodiazepin-2-one hydrochloride m.p. 126-145°C. lain +5.0°(0.100. CHC1_ dH (d6-DMSO) 8.20 (4H, d, J 9.2 Hz), 7.75-7.36 (9H, m), 7.08 (4H, d, J 9.2 Hz), 4.90 (IH, br s), 4.50 (4H, br s), 4.30-3.60 (5H, br m), and 3.34 (3H, s). Anal. Calcd. for C32H29N5θ7.HC1.0.15EtOH:
C, 60.71; H, 4.87; N, 10.96. Found: C, 60.70; H, 4.87; N, 10.70%.
EXAMPLE 63
Figure imgf000090_0002
(+)-3(R)-{ N-[3-(4-Nitrophenoxy)ethyl]amino } -1 ,3-dihydro-l -methy 1-
5-pheny 1-2H- 1.4-benzodiazepin-2-one hydrochloride m.p. 154-160°C, [α]D +84.6°(0.500, MeOH). dH (d6-DMSO) 10.2 (IH, br s), 8.25 (2H, d, J 9.0 Hz), 7.83-7.41 (9H, m), 7.09 (2H, d, J 9.0 Hz), 5.21 (IH, s), 4.57 (2H, m), 3.70 (2H, m),
3.47 (3H, s), and 3.40 (IH, m).
Anal. Calcd. for C24H22N4θ4.HC1.0.15EtOH.0.20H2θ:
C, 61.13; H, 5.13; N, 11.74. Found: C, 61.12; H, 4.92; N, 1 1.64%.
EXAMPLE 64
Figure imgf000091_0001
(+)-3(R)- { N-[3-(4-Chlorophenyl)prop- 1 -yl]amino } - 1 ,3-dihydro- 1 - methyl-5- phenyl-2H- 1.4-benzodiazepin-2-one hydrochloride m.p. 167-168°C, [α]D +20.8° (c=0.500, MeOH). dH (d6-DMSO) 9.9 (2H, br m), 7.78-7.26 (13H, m), 5.08 (IH, s), 3.45
(3H, s), 3.20 (IH, m), 3.00 (IH, m), 2.70 (2H, t, J 7.4 Hz), and 2.05
(2H, m).
Anal. Calcd. for C25H24CIN3O.HCI:
C, 66.08; H, 5.55; N, 9.25. Found: C, 65.81; H, 5.49; N, 9.30%. EXAMPLE 65
Figure imgf000092_0001
(+)-Phenylmethyl N-[(3R)-2,3-dmydro-l -me yl-5-phenyl-2-thioxo- lH-1.4-benzodiazepin-3-yllcarbamate
A mixture of (+)-pheny Imethy 1 N-[(3R)-2,3-dihydro-l- methyl-5-phenyl-2-oxo-lH-l ,4-benzodiazepin-3-yl]carbamate (4.0 g, 10 mmol) and 2,4-bis(4-methoxyphenyl)-l ,3-dithia-2,4-diphosphetane-2,4- disulfide (4.5 g, 1 1 mmol) in toluene (100 mL) was heated under reflux for 75 min. The mixture was cooled and the volume was reduced to 30 mL by evaporation under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with EtOAc/Hexane (75:25) to give (+)-phenylmethyl N-[(3R)-2,3-dihydro- 1 -methyl -5-phenyl-2-thioxo-lH-l,4-benzodiazepin-3-yl]carbamate as a solid, m.p. 128-131°C, [α]D +22.5° (c=0.656, CHCI3). dH (CDCI3) 7.65-7.26 (15H, m), 5.50 (IH, d, J 8.8 Hz), 5.14 (2H, s), and 3.86 (3H, s). Anal. Calcd. for C24H21N3O2S.O.25H2O:
C, 68.63; H, 5.16; N, 10.01. Found: C, 68.28; H, 5.21; N, 10.06%.
Employing the procedure substantially as described above, but substituting phenylmethyl N-[2,3-dihydro-5-phenyl-2- oxo-lH-l ,4-benzodiazepin-3-yl]carbamate for the (+)-phenylmethyl N-[(3R)-2,3-dihydro-l -methy l-5-phenyl-2-oxo-lH-l ,4-benzodiazepin- 3-yl]carbamate, the following compound was prepared: EXAMPLE 66
Figure imgf000093_0001
Phenyhnethyl N-[2,3-dihydro-5-phenyl-2-thioxo-l H- 1 ,4-benzodiazepin-
3-yllcarbamate dH (d6-DMSO) 10.85 (IH, s), 8.42 (IH, d, J 8.6 Hz), 7.65-7.10 (14H, m), 5.10 (2H, s), and 5.05 (IH, d, J 8.6 Hz).
EXAMPLE 67
Figure imgf000093_0002
3-Cyclohexyl-N-(2,3-dihydro- 1 -methyl-5-phenyl-2-thioxo- 1 H- 1 ,4- benzodiazepin-3-yPpropanamide
Hydrogen bromide was bubbled at room temperature through a solution of (+)-phenylmethyl N-[(3R)-2,3-dihydro-l-methyl- 5-phenyl-2-thioxo-lH-l ,4-benzodiazepin-3-yl]carbamate (0.9 g, 2.1 mmol), acetic acid (5 mL) and dichloromethane (5 mL). After 2 h., the solvent was evaporated under reduced pressure, ether was added and the solid was collected and dried in vacuo. A sample (0.58 g, 1.8 mmol) was suspended in THF (10 mL), triethylamine (0.24 mL, 0.18 g, 1.8 mmol) was added and the mixture was stirred at room temperature for 3 h. In a separate flask, oxalyl chloride (0.20 mL, 0.29 g, 2.3 mmol) was added to a solution of cyclohexanepropionic acid (0.33 mL, 0.30 g, 1.9 mmol) and DMF (1 drop) in THF (10 mL) and the mixture was stirred at room temperature for 3 h. The two mixtures were combined, triethylamine (0.32 mL, 0.23 g, 2.3 mmol) was added and the mixture was stirred at room temperature for 2.5 h. The solvent was evaporated under reduced pressure, water was added and the mixture was extracted with ethyl acetate. The combined organic fractions were washed with water, saturated aqueous sodium hydrogen carbonate, water (2 x) and brine, dried (Na2Sθ4) and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2θ2/MeOH (99.5:0.5) and the residue was recrystallized from EtOAc/Hexane to give 3-cyclohexyl-N-(2,3- dihydro- 1 -methy l-5-phenyl-2-thioxo- 1 H- 1 ,4-benzodiazepin-3- yl)propanamide as a solid, m.p. 219-221°C. dH (CDC13) 7.95 (IH, br d, J 8.6 Hz), 7.65-7.30 (9H, m), 5.72 (IH, d, J 8.6 Hz), 3.87 (3H, s), 2.41 (2H, t, J 7.6 Hz), and 1.80-0.85 (13H, m). Anal. Calcd. for C25H29N3OS.0.25H2O:
C, 70.81 ; H, 7.01; N, 9.91. Found: C, 70.80; H, 6.91; N, 9.95%.
Employing the procedure substantially as described above, but substituting phenylmethyl N-[2,3-dihydro-5-phenyl-2-thioxo-lH- l,4-benzodiazepin-3-yl]carbamate for the (+)-phenylmethyl N-[(3R)- 2,3-dihydro- 1 -methy 1-5 -pheny 1-2-thioxo- 1 H- 1 ,4-benzodiazepin-3- yljcarbamate and an appropriate acid for the cyclohexanepropionic acid, the following compounds were prepared: EXAMPLE 68
Figure imgf000095_0001
3-Cyclohexyl-N-(2,3-dihydro-5-phenyl-2-thioxo-lH-l,4-benzodiazepin-
3-yl)propanamide m.p. 1 13-1 19°C. dH (CDC13) 9.8 (IH, br s), 7.75-7.25 (10H, m), 5.75 (IH, d, J 8.1 Hz),
2.41 (2H, m), and 1.80-0.85 (13H, m).
Anal. Calcd. for C24H27N3OS.O.8CH2CI2:
C, 62.91 ; H, 6.09; N, 8.87. Found: C, 62.88; H, 5.70; N, 9.12%.
EXAMPLE 69
Figure imgf000095_0002
3-Cyclohexyl-N-(2,3-dihydro-2-hydrazono-5-phenyl-lH-l ,4- benzodiazepin-3v0propanamide
Hydrazine (53 mL, 56 mg, 1.8 mmol) was added to a solution of 3-cyclohexyl-N-(2,3-dihydro-l -methyl -5-phenyl-2-thioxo- lH-l ,4-benzodiazepin-3-yl)propanamide (120 mg, 0.25 mmol) in methanol (3 mL). The mixture was stirred at room temperature for 3 h. and the solvent was evaporated under reduced pressure. Ethyl acetate was added and the mixture was washed with water and brine, dried (Na2Sθ4) and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2Cl2/MeOH (99.5:0.5 increasing to 98:2) to give 3- cyclohexyl-N-(2,3-dihydro-2-hydrazono-5-phenyl- 1 H- 1 ,4-benzodiaze- pin-3yl)propanamide as a foam. dH (CDC13) 7.55-7.00 (1 IH, m), 5.75 (IH, d, J 7.6 Hz), 3.50 (2H, br s), 2.37 (2H, t, J 8.0 Hz), and 1.80-0.85 (13H, m). Anal. Calcd. for C24H29N5O.O.8CH3OH.O.I5CH2CI2:
C, 67.82; H, 7.41; N, 15.85. Found: C, 67.79; H, 7.46; N, 16.05%.
EXAMPLE 70
Figure imgf000096_0001
(E)- and (Z)-3-Cyclohexyl-N-(2,3-dihydro-2-hydroxyimino-5-phenyl-
1 H- 1.4-benzodiazepin-3-vDpropanamide
A mixture of 3-cyclohexyl-N-(2,3-dihydro-l-methyl-5- phenyl-2-thioxo-lH-l ,4-benzodiazepin-3-yl)propanamide (740 mg, 1.83 mmol), hydroxy lamine hydrochloride (140 mg, 2 mmol) and triethylamine (280 mL, 203 mg, 2 mmol) in methanol (15 mL)/THF (15 mL) was stirred at room temperature for 3 h. The solvent was evaporated under reduced pressure and the residue was purified by flash column chromatography on silica gel, eluting with CH2θ2/MeOH (98:2). The residue recrystallized from ethyl acetate. The first isomer to crystallize was recrystallized from ethyl acetate to give(E)-3-cyclohexyl-N-(2,3- dihydro-2-hydroxyimino-5-phenyl- IH- 1 ,4-benzodiazepin-3- yl)propanamide as a solid, m.p. 196°C. dH (d6-DMSO) 12.20 (IH, s), 9.00 (IH, d, J 8.0 Hz), 7.70-7.30 (10H, m), 5.45 (IH, d, J 8.0 Hz), 2.30 (2H, m), and 1.80-0.75 (13H, m).
The second isomer to crystallize was recrystallized from methanol to give (Z)-3-cyclohexyl-N-(2,3-dihydro-2-hydroxyimino-5- phenyl-lH-l ,4-benzodiazepin-3-yl)propanamide as a solid, m.p. 219°C. dH (d6-DMSO) 9.95 (IH, s), 8.95 (IH, s), 8.75 (IH, d, J 8.0 Hz), 7.50- 7.00 (9H, m), 5.70 (IH, d, J 8.0 Hz), 2.25 (2H, m), and 1.75-0.75 (13H, m). Anal. Calcd. for C24H28N4O2:
C, 71.26; H, 6.98; N, 13.85. Found: C, 70.89; H, 6.99; N, 13.55%.
EXAMPLE 71
Figure imgf000097_0001
3-Cyclohexyl-N-(2,3-dihydro- 1 -methyl-5-phenyl- 1 H- 1 ,4-benzodiazepin-
3yl) propanamide
Freshly prepared Raney nickel (400 mg) was added to a solution of 3-cyclohexyl-N-(2,3-dihydro-l -methyl -5-pheny I-2-thioxo- lH-l ,4-benzodiazepin-3-yl)propanamide (200 mg, 0.5 mmol) in ethanol (20 mL) and the mixture was stirred at room temperature for 2 h. The mixture was filtered and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2Cl2/MeOH (99.75:0.25) to give 3-cyclo- hexyl-N-(2,3-dihydro-l-methyl-5-phenyl-lH-l ,4-benzodiazepin-3yl) propanamide as a foam. dH (CDC13) 7.60-6.80 (9H, m), 6.37 (IH, br d, J 6.6 Hz), 5.53 (IH, m),
3.60 (2H, m), 2.77 (3H, s), 2.21 (2H, t, J 8.0 Hz), and 1.85-0.80
(13H, m).
Anal. Calcd. for C25H31N3O.O.2CH2CI2:
C, 74.45; H, 7.79; N, 10.34. Found: C, 74.68; H, 7.87; N, 10.23%.
EXAMPLE 72
1 -(2,3-Dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H-thieno-[2,3-e]- 1.4- diazepin-3-yD-3-(3-methyl-phenyl)urea
Step A:
Figure imgf000098_0001
(2-Amino-3-thienyl)phenylmethanone
Triethylamine (6.8 mL, 4.94 g, 49 mmol) was added to a heated (33°C) mixture of b-oxobenzenepropanenitrile (18.6 g, 128 mmol) and l,2-dithiane-2,5-diol (9.8 g, 64 mmol) in ethanol (120 mL) and the mixture was stirred at 50C° for 18 h. The mixture was cooled and the solvent was evaporated under reduced pressure. Dichloromethane was added, the mixture was washed with aqueous hydrochloric acid (0.5M), aqueous sodium hydroxide (IM) and brine, dried (Na2Sθ4) and the solvent was evaporated under reduced pressure. The residue was recrystallized from acetonitrile (150 mL) to give (2- amino-3-thienyl)-phenylmethanone as an orange solid (5.7 g, 44%). dH (CDC13) 7.70-7.35 (5H, m), 6.95 (2H, br s), 6.90 (IH, d, J 6.3 Hz), and 6.15 (lH, d, J 6.3 Hz).
Step B:
Figure imgf000099_0001
2.3-Dihydro-5-phenyl-lH-thienol2.3-el-1.4-diazepin-2-one
A solution of l,3-dihydro-l,3-dioxo-2H-isoindole-2-acetyl chloride (8.6 g, 38 mmol) in dichloromethane (20 mL) was added slowly to a cooled (0°C) mixture of (2-amino-3-thienyl)phenyl- methanone (6.8 g, 33 mmol), pyridine (6.34 mL, 6.20 g, 78 mmol) and 4-dimethylamino-pyridine (0.79 g, 6.5 mmol) in dichloromethane (130 mL). The mixture was stirred at 0°C for 30 min., diluted with dichloromethane (80 mL) and washed with aqueous hydrochloric acid (IM), saturated aqueous sodium hydrogen carbonate and brine. The mixture was dried (Na2Sθ4) and the solvent was evaporated under reduced pressure. The residue was triturated with ethanol and the solid was collected and dried in vacuo to give N-(3-benzoylthien-2-yl)-l,3- dihydro-l,3-dioxo-2H-isoindole-2-acetamide as a solid (9.8 g, 76%). A mixture of N-(3-benzoylthien-2-yl)-l,3-dihydro-l,3- dioxo-2H-isoindole-2-acetamide (10.9 g, 28 mmol) and hydrazine (1.9 mL, 1.94 g, 60 mmol) in THF (500 mL) was heated under reflux for 4 h. The mixture was cooled, filtered and the solvent was evaporated under reduced pressure. Saturated aqueous sodium hydrogen carbonate was added and the mixture was extracted with ethyl acetate. The combined organic fractions were washed with brine, dried (Na2Sθ4) and the solvent was evaporated under reduced pressure. Acetic acid (300 mL) was added and the mixture was heated under reflux for 15 min. The mixture was cooled and the solvent was evaporated under reduced pressure. Saturated aqueous sodium hydrogen carbonate was added and the mixture was extracted with ethyl acetate. The combined organic fractions were washed with brine, dried (Na2S04) and the solvent was evaporated under reduced pressure to give 2,3-dihydro-5- phenyl-lH-thieno[2,3-e]-l,4-diazepin-2-one as a foam (3.5 g, 52%). dH (CDC13) 9.75 (IH, br s), 7.90-7.30 (5H, m), 6.87 (IH, d, J 6.0 Hz), 6.82 (IH, d, J 6.0 Hz), and 4.45 (2H, s).
Step C:
Figure imgf000100_0001
2.3-Dihvdro-l-methyl-5-phenyl-lH-thienor2.3-el-L4-diazepin-2-one Sodium hydride (60% dispersion in mineral oil, 757 mg, 11.3 mmol) was added to a cooled (0°C) solution of 2,3-dihydro-5- phenyl-lH-thieno[2,3-e]-l,4-diazepin-2-one (2.61 g, 10.8 mmol) in DMF (7 mL). Further DMF (10 L) was added and the mixture was stirred for 30 min. A solution of iodomethane (0.67 mL, 1.53 g, 10.8 mmol) in ether (20 mL) was added and the mixture was stirred for 1 h. The mixture was poured into water and the mixture was extracted with ethyl acetate. The combined organic fractions were washed with brine, dried (Na2S04) and evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2Cl2/MeOH (95:5) to give 2,3-dihydro-l-methyl-5-phenyl-lH- thieno[2,3-e]-l,4-diazepin-2-one (1.5 g, 54%). dH (CDC13) 7.67-7.35 (5H, m), 7.00 (IH, d, J 6.0 Hz), 6.85 (IH, d, J 6.0 Hz), 4.45 (2H, br s), and 3.50 (3H, s).
Step D:
Figure imgf000101_0001
3-Amino-2,3-dihydro-l-methyl-5-phenyl-lH-thienof2,3-e]-l,4-diazepin- 2-one
2,3-Dihydro- 1 -methyl-5-pheny 1- 1 H-thieno[2,3-e] - 1 ,4- diazepin-2-one (1.5 g, 5.8 mmol) was dissolved in toluene (30 mL). The mixture was cooled to -10°C and potassium t-butoxide (1.7 g, 15.1 mmol) was added. The mixture was stirred at -10°C for 15 min., then isoamyl nitrite (1.0 mL, 0.87 g, 7.4 mmol) was added. The mixture was stirred at -10°C for 1 h. then allowed to warm to room temperature and poured into water (50 mL) and acetic acid (3 mL). The mixture was extracted with ethyl acetate and the combined organic fractions were washed with brine, dried (Na2Sθ4) and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with EtOAc/Hexane to give 2,3- dihydro- 1 -methyl-3-hydroxyimino-5-phenyl- 1 H-thieno[2,3-e]-l ,4- diazepin-2-one (0.80 g, 48%).
2,3-Dihydro- 1 -methyl-3-hydroxyimino-5-phenyl- 1 H-thieno [2,3-e]-l,4-diazepin-2-one (0.80 g, 2.8 mmol) was dissolved in ethanol (40 mL) and Raney nickel (2 g) was added. The mixture was shaken under hydrogen (50 p.s.i.) for 5 days, adding further Raney nickel (10 g) in portions. The mixture was filtered and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2θ2/MeOH to give 3- amino-2,3-dihydro-l-methyl-5-phenyl-lH-thieno[2,3-e]-l,4-diazepin-2- one (248 mg, 33%). dH (CDC13) 7.50-7.30 (5H, m), 7.05 (IH, d, J 6.0 Hz), 6.85 (IH, d, J
6.0 Hz), 4.57 (IH, s), 3.55 (3H, s), and 1.70 (2H, br s).
Step E:
Figure imgf000102_0001
1 -(2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H-thieno [2,3-e] - 1 ,4-diazepin- 3-ylV3-(3-methylphenyOurea
3-Methylphenylisocyanate (60 mL, 62 mg, 0.46 mmol) was added to a solution of 3-amino-2,3-dihydro-l-methyl-5-phenyl-lH- thieno[2,3-e]-l,4-diazepin-2-one (124 mg, 0.46 mmol) in tetrahydro- furan (5 mL). The mixture was stirred at room temperature for 2 h. and the solvent was evaporated under reduced pressure. The residue was crystallized from EtOAc (4 mL) to give l-(2,3-dihydro-l-methyl- 2-oxo-5-phenyl-lH-thieno[2,3-e]-l,4-diazepin-3-yl)-3-(3-methyl- phenyl)urea as a solid (94 mg, 50%). m.p. 128-130°C. dH (CDC13) 8.70 (IH, s), 7.65-6.75 (12H, m), 5.55 (IH, d, J 9.0 Hz), 3.55 (3H, s), and 2.30 (3H, s). Anal. Calcd. for C22H20N4O2S.O.25H2O:
C, 64.62; H, 4.99; N, 13.70. Found: C, 64.68; H, 4.96; N, 13.70%. EXAMPLE 73
Figure imgf000103_0001
3-Cyclohexyl-N-(2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH-thieno[2,3- el-1.4-diazepin-3-vPpropanamide
Triethylamine (75 mL, 54 mg, 0.54 mmol) was added to a mixture of 3-amino-2,3-dihydro-l-methyl-5-phenyl lH-thieno[2,3-e]- 1 ,4-diazepin-2-one (82 mg, 0.3 mmol), cyclohexanepropanoic acid (52 mL, 47 mg, 0.3 mmol), l-(3-dimethylaminopropyl)-3-ethylcarbodi- imide hydrochloride (58 mg, 0.3 mmol) and 1-hydroxybenzotriazole (42 mg, 0.3 mmol) in DMF (1.5 mL). The mixture was stirred at room temperature for 18 h. and ethyl acetate (60 mL) was added. The mixture was washed with aqueous citric acid (10%), saturated aqueous sodium hydrogen carbonate and brine, dried (Na2S04) and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with EtOAc/Hexane to give 3-cyclohexyl-N-(2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH- thieno[2,3-e]-l ,4-diazepin-3-yl)propanamide as a solid (56 mg, 46%). m.p. 189-190°C. dH (CDC13) 7.65-6.85 (8H, m), 5.65 (IH, d, J 8.0 Hz), 3.55 (3H, s), 2.40 (2H, t, J 7.0 Hz), and 1.80-0.85 (13H, m). Anal. Calcd. for C23H27N3O2S.O.5H2O:
C, 66.00; H, 6.74; N, 10.04. Found: C, 66.25; H, 6.76; N, 9.83%. EXAMPLE 74
Figure imgf000104_0001
3-Cyclohexyl-N-(5-cyclohexyl-2,3-dihydro-2-oxo-lH-l ,4- benzodiazepin-3-vπ propanamide
Phenylmethyl N-[5-cyclohexyl-2,3-dihydro-2-oxo-lH-l ,4- benzodiazepin-3-yl]carbamate (150 mg, 0.38 mmol) was dissolved in hydrogen bromide in acetic acid (30%, 0.5 mL). After 2 h., ether was added and the solid was collected and dried in vacuo. THF (3 mL) and triethylamine (0.45 mL, 33 mg, 0.32 mmol) were added and the mixture was stirred at room temperature for 3 h. In a separate flask, oxalyl chloride (38 mL, 56 mg, 0.44 mmol) was added to a solution of cyclohexanepropionic acid (61 mL, 56 mg, 0.36 mmol) and DMF (1 drop) in THF (2 mL) and the mixture was stirred at room temperature for 3 h. The two mixtures were combined, triethylamine (61 mL, 44 mg, 0.44 mmol) was added and the mixture was stirced at room temperature for 3 h. The solvent was evaporated under reduced pressure and ethyl acetate was added. The mixture was washed with water (2 x), saturated aqueous sodium hydrogen carbonate, water and brine, dried (Na2Sθ4) and the solvent was evaporated under reduced pressure. The residue was recrystallized from i-PrOH to give 3- cyclohexyl-N-(5-cyclohexy l-2,3-dihydro-2-oxo- 1 H- 1 ,4-benzodiazepin- 3-yl)propanamide as a solid, m.p. 133-138°C. dH (CDC13) 7.85 (IH, br s), 7.62-6.95 (5H, m), 5.40 (IH, d, J 8.7 Hz), 2.77 (IH, m), 2.34 (2H, m), and 2.05-0.75 (23H, m). Anal. Calcd. for C24H33N3O2.O.7C3H7OH:
C, 71.64; H, 8.89; N, 9.60. Found: C, 71.28; H, 8.70; N, 9.82%.
EXAMPLE 75
Figure imgf000105_0001
(+)-N-[(3R)-7-Amino-2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4- benzodiazepm-3-yl1-3-(2.4-dichlorophenvDpropanamide
Step A:
To a mixture of 3(R)-amino-l ,3-dihydro-l-methyl-5- phenyl-2H-l ,4-benzodiazepin-2-one (J. Org. Chem. 1987, 52, 3232- 3239) (3.98 g, 15.0 mmol) in concentrated sulfuric acid (15 mL) cooled in an ice-bath was added dropwise a solution of potassium nitrate (2.12 g, 21.0 mmol) in concentrated sulfuric acid (6 mL). The mixture was stirred with cooling for 2 h., then stirred at ambient temperature for 1.5 h. Ice (80 g) was added and the mixture was basified with concentrated ammonium hydroxide to pH 9. The resulting mixture was extracted with ethyl acetate (3 x 220 mL). The combined organic fractions were washed with brine, dried (Na2S04) and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with chloroform/methanol (97:3). The material which eluted was further purified by flash column chromatography on silica gel, eluting with ethyl acetete/methanol (95:5). The material which eluted was stirred under w-butyl chloride (30 mL) and the solvent was evaporated under reduced pressure to give an inseparable mixture of 3(R)-amino-l ,3-dihydro-l -methyl-7-nitro-5- phenyl-2H-l ,4-benzodiazepin-2-one and 3(R)-amino-l ,3-dihydro-l - methyl-7-nitro-5-(2-nitrophenyl)-2H-l ,4-benzodiazepin-2-one (3.81 g) in a 3:1 ratio as a yellow solid. dH (CDCI3) (mononitro compound) 8.43 (IH, dd, J 9, 3 Hz), 8.23 (IH, d, J 3 Hz), 7.59 (2H, m), 7.52 (2H,m), 7.44 (2H,m), 4.47 (lH,s), 3.53 (3H,s), and 2.42 (2H, br s); (dinitro compound) 8.49 (IH, dd, J 9, 3), 8.42 (IH, m), 8.18 (IH, d, J 3 Hz), 8.01 (IH, m), 7.67 (IH, t, J 6 Hz), 7.6-7.4 (2H, m), 4.52 (lH,s), 3.56 (3H,s), and 2.42 (2H, br s).
Step B:
A solution of 3-(2,4-dichlorophenyl)propionic acid (482 mg, 2.2 mmol), DMF (0.017 mL, 0.22 mmol), and thionyl chloride (0.24 mL, 3.3 mmol) in chloroform (2.5 mL) was heated at reflux for 1 h. The solvent was evaporated under reduced pressure to give 3-(2,4- dichlorophenyl)propionyl chloride (520 mg, 100%). To a solution of mixed 3(R)-amino- 1 ,3-dihydro- 1 -methy l-7-nitro-5-phenyl-2H-l ,4- benzodiazepin-2-one and 3(R)-amino-l ,3-dihydro-l -methyl-7-nitro-5- (2-nitrophenyl)-2H-l,4-benzodiazepin-2-one (3:1) (621 mg, 2 mmol) and triethylamine (0.305 mL, 2.2 mmol) in methylene chloride (10 mL), was added a solution of 3-(2,4-dichlorophenyl)propionyl chloride (520 mg, 2.2 mmol) in methylene chloride (1.5 mL). The mixture was stirred for 30 min., the solvent was partially evaporated under reduced pressure, and the reaction mixture was purified by flash column chromatography on silica gel, eluting with methylene chloride/ether (90:10) to give a mixture of (+)-N-[(3R)-2,3-dihydro-l-methyl-7-nitro- 2-oxo-5-phenyl-lH-l ,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)- propanamide and (+)-N-[(3R)-2,3-dihydro-l-methyl-7-nitro-2-oxo-5- (2-nitrophenyl)-l H- 1 ,4-benzodiazepin-3-yl]-3-(2,4-dichloroρhenyl)- propanamide (850 mg, 84%) in a 3:1 ratio as a solid white foam. dH (CDCI3) (mononitro compound) 8.45 (IH, dd, J 9, 3 Hz), 8.25 (IH, d J 3 Hz), 7.54 (3H, m), 7.45 (2H, m), 7.38 (IH, d, J 2 Hz), 7.26-7.18 (4H, m), 5.50 (IH, d, J 8 Hz), 3.52 (3H, s), 3.10 (2H, m), and 2.70 (2H, m); (dinitro compound) 8.51 (IH, dd, J 9, 3 Hz), 8.40 (IH, m), 8.21 (IH, d J 3 Hz), 7.98 (IH, m), 7.68 (IH, t, J 6 Hz), 7.60 (IH, m), 7.44 (1H, m), 7.26-7.15 (4H, m), 5.52 (IH, d, J 8 Hz), 3.55 (3H, s), 3.10 (2H, m), and 2.70 (2H, m).
Step C:
To a solution of mixed N-[(3R)-2,3-dihydro-l -methyl- 7-nitro-2-oxo-5-phenyl-lH-l ,4-benzodiazepin-3-yl]-3-(2,4-dichloro- pheny l)propanamide and (+)-N-[(3R)-2,3-dihydro- 1 -methyl-7-nitro-2- oxo-5-(2-nitrophenyl)-lH- l ,4-benzodiazepin-3-yl]-3-(2,4-dichloro- phenyOpropanamide (3:1) (770 mg, 1.5 mmol) in acetic acid (6 mL) was added dropwise in portions over 1.5 h. a solution of 15% titanium (ID) chloride in 20-30% hydrochloric acid (7.8 mL, 9.0 mmol). The resulting solution was stirred 30 min., basified with 20% sodium hydroxide solution (pH 9), diluted with water (80 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic fractions were washed with brine, dried (Na2S04) and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with ethyl acetare/hexane (75:25 increasing to 100:0). The first compound to elute was crystallized from ethyl acetate to give (+)-N-[(3R)-7-amino-2,3-dihydro-l-methyl-2-oxo- 5-phenyl-lH-l ,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)- propanamide (413 mg, 57%) as a pale yellow solid, m.p. 179-180°C, [cc]D +60.2° (c= 0.500, CHC13). dH (CDCI3) 7.60 (2H, d, J 7 Hz), 7.49-7.36 (5H, m) 7.24 (IH, d, J 9 Hz), 7.17 (2H, m), 6.99 (IH, dd, J 9, 3 Hz), 6.64 (lH,d, J 3 Hz), 5.54 (IH, d, J 8 Hz), 4.80-3.50 (2H, br s), 3.39 (3H, s), 3.09 (2H, t, J 8 Hz), and 2.68 (2H, dt, Jd 3, Jt 8 Hz). Anal. Calcd. for C25H22CI2N4O2:
C, 62.38; H, 4.61; N, 11.64. Found: C, 62.58; H, 4.68; N, 11.65%.
The second compound to elute was crystallized from ethyl acetate to give (+)-N-[(3R)-7-amino-2,3-dihydro-l-methyl-2-oxo-5- (2-aminophenyl)-lH-l ,4-benzodiazepin-3-yl]-3-(2,4-dιchlorophenyl)- propanamide (114 mg, 15%) as a pale yellow solid, m.p. 188-189°C, [α]D +50.0° (c=0.100, MeOH). dH (CDC13) 7.36 (2H, m), 7.25 (IH, d, J 9 Hz), 7.15 (3H, m), 7.00 (IH, m), 6.88 (2H, m), 6.79 (IH, m), 6.60 (IH, bs), 5.52 (IH, d, J 8 Hz),
4.10-2.80 (4H br s), 3.40 (3H, m), 3.09 (2H, t, J 8 Hz), and 2.69 (2H, m).
Anal. Calcd. for C25H23Cl2N5θ2-0.05EtOAc:
C, 60.43; H, 4.71 ; N, 13.99. Found: C, 60.79; H, 4.74; N, 13.83%.
EXAMPLE 76
Figure imgf000108_0001
(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl-7-methane- sulfonamido-lH-l ,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)- propanamide
Methanesulfonyl chloride (0.040 mL, 0.52 mmol) was added to a solution of (+)-N-[(3R)-7-amino-2,3-dihydro-l-methyl-2- oxo-5-phenyl-lH-l,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)- propanamide (193 mg, 0.40 mmol) and pyridine (0.065 mL, 0.80 mmol) in methylene chloride (1.6 mL). The resulting solution was stirred 2 h. The solution was diluted with ethyl acetate (12 mL), washed with IN HC1, water, saturated sodium bicarbonate solution, water, and brine (3 mL each), dried (Na2S04) and the solvent was evaporated under reduced pressure. The residue was dissolved in warm toluene, treated with charcoal, and filtered. The filtrate was diluted with hexane, the mixture was cooled, and the resulting precipitate was collected and dried in vacuo to give (+)-N-[(3R)-2,3-dihydro-l-methyl-2-oxo-5- phenyl-7-methanesulfonamido- 1 H- 1 ,4-benzodiazepin-3-yl]-3-(2,4- dichloropheny propanamide (152 mg, 68%) as a white solid, m.p. 130- 148°C, [α]D +11 1.6° (c=0.500, CHCI3). dH (CDCI3) 7.55-7.32 (9H, m), 7.24 (2H, dd, J 10, 2 Hz), 7.17 (IH, dd, J 9, 2 Hz), 7.05 (IH, d, J 3 Hz), 5.49 (IH, d, J 8 Hz), 3.41 (3H, s), 3.08 (2H, t, J 8 Hz), 2.97 (3H, s), and 2.71 (2H, dt, Jd 3, Jt 8 Hz). Anal. Calcd. for C26H24θ2N4θ4S:
C, 55.82; H, 4.32; N, 10.01. Found: C, 56.12; H, 4.47; N, 9.89%.
EXAMPLE 77
Figure imgf000109_0001
N-(2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H-pyrido[4,3-e]- 1 ,4- diazepin-3-vD-3-f2.4-dichlorophenyDpropanamide hydrochloride
Step A:
To a solution of 2,3 -dihydro- 1 -methy 1-5-pheny 1-1 H- pyrido[4,3-e]-l,4-diazepine-2-one (J. Med. Chem.. 1965, 8, 722-724) (1.63 g, 6.5 mmol) in toluene (32 mL) under argon cooled to -20°C (ice/methanol bath) was added potassium t-butoxide (1.83 g, 16.3 mmol). The resulting purple suspension was stined 15 min. at -20°C and isoamyl nitrite (1.05 mL, 7.8 mmol) was added. The mixture was stirred at -20°C for 30 min., then poured into a mixture of water (50 mL), acetic acid (3 mL), and ethyl acetate (65 mL). The mixture was stirred to dissolve all solids and the layers were separated. The aqueous layer was extracted with ethyl acetate (65 mL). The combined organic fractions were washed with saturated sodium bicarbonate solution and brine (20 mL each), dried (Na2S04), and the solvent was evaporated under reduced pressure. The residue was triturated with cold toluene and the solid was collected and dried in vacuo to give 2,3-dihydro-3- hydroxyimino- 1 -methyl-5-phenyl- 1 H-pyrido[4,3-e]- 1 ,4-diazepine-2-one (1.22 g, 67%) as a yellow solid, m.p. 223-224°C. dH (CDC13) 8.92 (IH, bs), 8.73 (IH, d, J 7 Hz), 8.62 (IH, s), 7.80 (2H, dd, J 7, 1 Hz), 7.59 (IH, m), 7.48 (2H, ), 7.26 (IH, d, J 7 Hz), and 3.50 (3H,s).
Step B:
A mixture of 2,3-dihydro-3-hydroxyimino-l-methyl-5- phenyl-lH-pyrido[4,3-e]-l,4-diazepine-2-one (1.77 g, 6.3 mmol) and freshly prepared Raney nickel (3.2 g) in 1 :1 ethanol/methanol (190 mL) was shaken on a Parr hydrogenation apparatus under hydrogen (50 psi) for 4 h. The mixture was filtered through filter aid and the filtrate was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with methanol/ chloroform/acetic acid (5:95: 1 increasing to 10:90:1 ). The material which eluted was stirred under chloroform (30 mL) with potassium carbonate (0.3 g) and water (0.2 mL) for 5 min. The mixture was dried (Na2Sθ4) and the solvent was evaporated under reduced pressure to give 3-amino-2,3-dihydro-l -methyl-5-phenyl-lH-pyrido[4,3-e]-l ,4- diazepine-2-one (276 mg, 16%), as a yellow solid, m.p. 109-123°C. dH (CDC13) 8.72 (IH, d, J 6 Hz), 8.58 (IH, s), 7.61 (2H, m), 7.51 (IH, m), 7.43 (2H, m), 7.26 (IH, m), 4.47 (IH ,s), 3.50 (3H, s), and 2.1 (2H, bs).
High res. mass spectrum: Theoretical mass for C15H14N4O (M+l): 267.124586. Measured mass (M+l): 267.123654. Step C:
A solution of dicyclohexylcarbodiimide (87 mg, 0.42 mmol) in methylene chloride (0.17 mL) was added to a solution of 3- amino-2,3-dihydro-l -methyl-5-phenyl-lH-pyrido[4,3-e]-l ,4-diazepine- 2-one (93 mg, 0.35 mmol) and 3-(2,4-dichlorophenyl propionic acid (83 mg, 0.38 mmol) in tetrahydrofuran (0.5 mL) under argon. The resulting mixture was stirred for 5 h., filtered, and the filtrate was evaporated under reduced pressure. The residue was purified by preparative plate chromatography on silica gel eluting with methanol/ chloroform/acetic acid (5:95:1). The purified material was stirred under chloroform (5 mL) with potassium carbonate (0.1 g) and water (2 drops) for 5 min. The mixture was dried (Na2Sθ4) and the solvent was evaporated under reduced pressure. The residue was suspended in ethanol (2 mL) and ethanolic HC1 (6.8 M, 0.147 mL) was added. The mixture was stined, the resulting precipitate was collected and dried in vacuo to give N-(2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH-pyrido[4,3- e]-l ,4-diazepin-3-yl)-3-(2,4-dichlorophenyl)propanamide hydrochloride (32 mg, 18%) as a white solid, m.p. 218-219°C. dH (d6-DMSO) 9.38 (IH, d, J 8 Hz), 8.86 (IH, bs), 8.59 (IH bs), 7.79 (IH, d, J 6 Hz), 7.56 (3H, m), 7.51 (2H, m), 7.39 (2H, m), 7.25 (IH, m), 7.16 (IH, m), 5.37 (IH, d, J 8 Hz), 3.44 (3H, s) 2.94 (2H, t, J 7 Hz), and 2.64 (2H, t, J 7 Hz). Anal. Calcd. for C24H20CI2N4O2.HCI:
C, 57.22; H, 4.20; N, 1 1.12. Found: C, 56.87; H, 4.18; N, 11.09%.
EXAMPLE 78
Figure imgf000112_0001
N-(2,3-Dihydro-l -methyl -2-oxo-5 -phenyl- 1 H-pyrido[4,3-el- 1 ,4- diazepin-3-yO-3-(cvclohexyDpropanamide
A solution of dicyclohexylcarbodiimide (87 mg, 0.42 mmol) in methylene chloride (0.17 mL) was added to a solution of 3- amino-2,3-dihydro-l-methyl-5-phenyl-lH-pyrido[4,3-e]-l ,4-diazepine- 2-one (93 mg, 0.35 mmol) and cyclohexanepropionic acid (0.065 mL, 0.38 mmol) in tetrahydrofuran (0.5 mL) under argon. The resulting mixture was stirred for 5 h., filtered, and the filtrate was evaporated under reduced pressure. The residue was purified by preparative plate chromatography on silica gel eluting with methanol/chloroform/acetic acid (5:95:1). The purified material was stirred under chloroform (5 mL) with potassium carbonate (0.1 g) and water (2 drops) for 5 min. The mixture was dried (Na2Sθ4) and the solvent was evaporated under reduced pressure. The residue was crystallized from toluene to give N- (2,3-dihydro-l -methy l-2-oxo-5-phenyl-lH-pyrido[4,3-e]-l,4-diazepin-3- yl)-3-(cyclohexyl)-propanamide (47 mg, 33%) as a white crystalline solid, m.p. 170-173°C. dH (CDC13) 8.75 (IH, d, J 6 Hz), 8.61 (IH, s), 7.58 (2H, m), 7.52 (IH, m), 7.45 (2H, m), 7.31 (IH, d, J 6 Hz), 7.21 (IH, d, J 8 Hz), 5.54 (IH, d, J 8 Hz), 3.51 (3H, s), 2.39 (2H, m), 1.73 (4H, m), 1.63 (3H, m), 1.85- 1.12 (4H, m), and 0.94 (2H, m). Anal. Calcd. for C24H28N4θ2.0.10PhCH3:
C, 71.70; H, 7.02; N, 13.54. H I
Found: C, 71.78; H, 7.01; N, 13.57%.
Employing the procedure substantially as described above, but substituting 3-(4-trifluoromethylphenyl)-propionic acid for the cyclohexanepropionic acid, the following compound was prepared:
EXAMPLE 79
Figure imgf000113_0001
N-(2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H-pyrido[4,3-e]- 1 ,4- diazepin-3-yl)-3-(4-trifluoromethylphenyl propanamide m.p. 191-192°C. dH (CDC13) 8.76 (IH, d, J 6 Hz), 8.61 (IH, s), 7.56 (4H, m), 7.52 (IH, m), 7.42 (2H, d, J 7 Hz), 7.38 (2H, m), 7.30 (IH, d, J 6 Hz), 7.22 (IH, d, J 8 Hz), 5.51 (IH, d, J 8 Hz), 3.50 (3H, s), 3.09 (2H, t, J 8 Hz), and
2.73 (2H, t, J 8 Hz).
Anal. Calcd. for C25H2lF3N4θ2.0.20PhCH3:
C, 65.39; H, 4.70; N, 11.56. Found: C, 65.69; H, 4.64; N, 1 1.95%. EXAMPLE 80
Figure imgf000114_0001
N-(2,3-Dihydro-l-memyl-2-oxo-5-phenyl-lH-pyrido[3,4-e]-l,4- diazepin- 3 -y 1 )-3 -(2 ,4-dichloropheny 1 )propanamide
Step A:
To a solution of 2,3-dihydro-l -methyl-5-phenyl-lH- pyrido[3,4-e]-l ,4-diazepine-2-one (Can. J. Chem. 1987, 65, 1158-1161) (1.43 g, 5.7mmol) in toluene (28 mL) under argon cooled to -20°C (ice/methanol bath) was added potassium t-butoxide (1.59 g, 14.2 mmol). The resulting purple suspension was stirred 15 min. at -20 °C and isoamyl nitrite (0.92 mL, 6.8 mmol) was added. The mixture was stirred at -20°C for 30 min., then poured into a mixture of water (25 mL), acetic acid (2.5 mL), and ethyl acetate (55 mL). The mixture was stirred to dissolve all solids and the layers were separated. The aqueous layer was extracted with ethyl acetate (2 x 55 mL). The combined organic fractions were washed with saturated sodium bicarbonate solution and brine (20 mL each), dried (Na2Sθ4), and the solvent was evaporated under reduced pressure. The residue was triturated with hexane and the solid was collected and dried in vacuo to give 2,3- dihydro-3-hydroxyimino- 1 -methyl-5-phenyl- 1 H-pyrido[3,4-e]-l ,4- diazepine-2-one (1.60 g, 100%) as a tan foam. dH (CDC13) 8.77 (IH, s), 8.50 (IH, d, J 4 Hz), 7.81 (2H, dd, J 8, 1 Hz), 7.60 (IH, m), 7.49 (3H, m), 7.32 (IH, d, J 5 Hz), and 3.55 (3H,s). Step B:
A solution of stannous chloride dihydrate (3.72 g, 16.5 mmol) in concentrated hydrochloric acid (11 mL) was added dropwise to 2,3-dihydro-3-hydroxyimino-l-methyl-5-phenyl-lH-pyrido[3,4-e]- 1 ,4-diazepine-2-one (1.54 g, 5.5 mmol) cooled in an ice bath. The resulting solution was stirred at ambient temperature for 3 h. The solution was diluted with water (20mL), basified with concentrated ammonium hydroxide (18 mL), and extracted with ether (4 x 75 mL). The combined organic fractions were washed with brine (30 mL), dried (Na2S04), and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with methanol/chloroform/acetic acid (5:95: 1 increasing to 10:90: 1 ). The material which eluted was stirred under chloroform (20 mL) with potassium carbonate (0.3 g) and water (2 drops) for 5 min. The mixture was dried (Na2S04) and the solvent was evaporated under reduced pressure. The residue was stirred under hexane, and the resulting solid was collected to give 3-amino-2,3-dihydro-l -methyl-5- phenyl-lH-pyrido[3,4-e]-l,4-diazepine-2-one (241 mg, 16%) as a yellow solid, m.p. 94-118°C. dH (CDC13) 8.79 (IH, s), 8.48 (IH, d, J 5 Hz), 7.62 (2H, dd, J 8, 1 Hz), 7.51 (IH, m), 7.45 (2H, m), 7.24 (IH, dd, J 5, 1 Hz), 4.47 (IH ,s), 3.55 (3H, s), and 2.2 (2H, bs). Anal. Calcd. for Cl5Hl4N4θ.0.25(C_H5)2θ:
C, 67.46; H, 5.84; N, 19.67. Found: C, 67.28; H, 5.66; N, 19.53%.
High res. mass spectrum: Theoretical mass for C15H14N4O (M+l): 267.124586. Measured mass (M+l): 267.123093.
Step C:
A solution of oxalyl chloride (0.023 mL, 0.26 mmol) in methylene chloride (0.2 mL) was added dropwise to a solution of 3- (2,4-dichlorophenyl)propionic acid (48 mg, 0.22 mmol) and DMF (1 drop) in methylene chloride (0.5 mL) cooled in an ice-bath. The resulting solution was stirred 1 h. with cooling. The solvent was evaporated under reduced pressure to give 3-(2,4-dichlorophenyl)- propionyl chloride (52 mg, 100%). To a solution of 3-amino-2,3- dihydro-l-methyl-5-phenyl-lH-pyrido[3,4-e]-l ,4-diazepine-2-one (53 mg, 0.20 mmol) and pyridine (0.021 mL, 0.22 mmol) in methylene chloride (3 mL), was added a solution of 3-(2,4-dichlorophenyl)- propionyl chloride (52 mg, 0.22 mmol) in methylene chloride (0.5 mL). The mixture was stirred for 1 h., the solvent was partially evaporated under reduced pressure, and the reaction mixture was purified by flash column chromatography on silica gel, eluting with methanol/ether (5:95 increasing to 7.5:92.5). The material which eluted was crystallized from toluene/hexane to give N-(2,3-dihydro-l-methyl-2-oxo-5-phenyl- lH-pyrido[3,4-e]-l ,4-diazepin-3-yl)-3-(2,4-dichlorophenyl)propanamide (38 mg, 38%) as a white crystalline solid, m.p. 220-22PC. dH (CDC13) 8.81 (IH, s), 8.52 (IH, d, J 5 Hz), 7.56 (2H, dd, J 7, 2 Hz), 7.51 (IH, m), 7.44 (2H, d, J 6 Hz), 7.40 (IH, m), 7.27 (2H, m), 7.18 (2H, dd, J 8, 2 Hz), 5.48 (IH ,d, J 8 Hz), 3.55 (3H, s), 3.10 (2H, t, J 7 Hz), and 2.71 (2H, dt, Jd 2 Jt 8 Hz). Anal. Calcd. for C24H20Cl2N4θ2.0.25PhCH3:
C, 63.06; H, 4.52; N, 11.43. Found: C, 63.03; H, 4.48; N, 11.25%.
EXAMPLE 81
N-[2,3-Dihydro-l -memyl-2-oxo-5-isopropyl-lH-l ,4-benzodiazepin-3- yll-3-(2.4-dichlorophenyDpropanamide
Step A:
Figure imgf000116_0001
To a solution of the benzodiazepine (1.0 g, 5.3 mmol) in THF (20 mL) at -78°C under argon was added 60% (NaH, 2.52 g, 6.3 mmol) Boc anhydride (1.27 g, 5.8 mmol) and the mixture stirred at -78°C for 1/2 hour. The reaction was then allowed to warm to 25°C and stirred for 2 hours before quenching into cold aq. NH4CI (10%) and extracting the product into ethyl acetate (3x50 mL). Concentration of the dried (Na2S04) extracts gave an oil which was passed through silica (EtOAc/hexane) to give 1.35 g product (89%). IH NMR (CDCI3) d: 1.60 (s, 9H), 3.40 (s, 3H), 3.95 (brd, IH), 4.80 (brd, IH), 7.20 (d, IH), 7.30 (q, IH), 7.60 (t, IH), 7.92 (d, IH).
Step B:
Figure imgf000117_0001
To a solution of the BOC-benzodiazepine (4.0 g, 13.8 mmol) in THF (80 mL) under argon was rapidly added a solution of isopropylmagnesium chloride (2.0 M) in THF (7.66 mL, 15.3 mmol). The reaction was stirred for 1/2 hour, quenched into aq NH4CI (50 mL), and extracted with ethyl acetate (2x200 mL). The organic extracts were concentrated and chromatographed on silica (1 :1, EtOAC/hexane) to give 1.55 g (34%) of product.
IH NMR (CDCI3) d: 1.14 (d, 3H), 1.19 (d, 3H), 1.40 (s, 9H), 3.13 (s, 3H), 3.2-3.8 (m, 3H), 5.45 (brs, IH), 7.28 (dt, IH), 7.48 (dt, IH), 7.56 (dt, IH), 7.72 (dd, IH). Step C:
Figure imgf000118_0001
To a 0°C solution of the isopropylphenone (1.55 g) in ethyl acetate was added anhydrous HCl gas over 90 min. The reaction was then concentrated jn vacuo to give a solid which was dissolved in H2θ (40 mL) and the pH adjusted to 11.0 with IN LiOH. After 30 min. at pH = 11.0 the pH was adjusted to 7.0 with IN HCl and product extracted into ethyl acetate. The organic extracts were dried (Na2Sθ4), filtered and concentrated to give a solid 1.22 g, 100%.
IH NMR (CDCI3) d: 0.95 (d, 3H), 1.30 (d, 3H), 3.16 (septet, I H), 3.36
(s, 3H), 3.60 (d, IH), 4.60 (d, IH), 7.2-7.3 (m, 2H), 7.45-7.55 (m, 2H).
Step D:
The benzodiazepine obtained in Step C was converted to the oxime as described in Example 80 Step A.
Step E:
The oxime (2 gms) was dissolved in acetic acid (150 mL) and 10% Pd/C (1 gm) added. The mixture was stirred rapidly under an atmosphere of hydrogen for 90 min or until complete by HPLC. The reaction was filtered, the catalyst washed with methylene chloride (200 mL) and the filtrates concentrated in vacuo to an oil. The oil was dissolved in saturated aqueous sodium bicarbonate (100 mL) and product extracted with ethyl acetate (3 x 150 mLs). Concentration of the dried (Na2Sθ4) extracts gave 2.60 gms (97%).
Step F: The anine was coupled with 3-(2,4-dichlorophenyl)- propionic acid as described in Example 43 to yield N-(2,3-dihydro-l- methyl-2-oxo-5-isopropyl-lH-l ,4-benzodiazepin-3-yl)-3-(2,4- dichlorophenyl)propanamide.
IH NMR (CDC13) d: 0.92 (d, 3H), 1.25 (d, 3H), 2.65 (dt, 2H), 3.05 (t, 2H), 3.15 (SepT, IH), 3.40 (s, 3H), 5.38 (d, IH), 7.0-7.6 (m, 8H).
The following compounds were prepared in a similar manner as described in Example 81, using the appropriate Grignard reagent in place of isopropyl magnesium chloride.
EXAMPLE 82
N-[2,3-dihydro-l -me yl-2-oxo-5-isoproρyl-lH-l,4-benzodiazepin-3- yll-3-cvclohexylpropanamide m.p. 164-165°C
CHN: Anal. Calcd. for C22H31N3O2:
C, 71.51; H, 8.46; N, 1 1.37 Observed: C, 71.72; H, 8.39; N, 11.32
EXAMPLE 83
N-[2,3-dihydro-l-metnyl-2-oxo-5-isopropyl-lH-l,4-benzodiazepin-3- yll-3-(4-trifluoromethylphenvDpropanamide m.p. 187-188°C
IH NMR (CDCI3) d: 0.92 (d, 3H), 1.25 (d, 3H), 2.66 (dt, 2H), 3.04 (t, 2H), 3,15 (SepT, IH), 3.40 (S, 3H), 5.38 (d, IH), 7.14 (brd, IH), 7.25- 7.6 (m, 8H).
Employing substantially the same methods described in Example 80, but replacing Step E with the reduction method described below, the following compounds were prepared:
Figure imgf000120_0001
To a solution of the oxime i (1.28 g, 0.0048 mole) in H2θ (130 ml) and THF (65 ml) was added sodium dithionite (Na2S2θ4) (13.0 g, 0.075 mole). The mixture was stirred for 2 hours then diluted with saturated aqueous sodium bicarbonate (50 ml) and product extracted into ethyl acetate (2 x 150 ml). The organic extracts were combined, dried over Na2Sθ4, filtered, and concentrated to give an oil ( 1.0 g). The oil was chromatographed on silica using ethyl acetate followed by 10% methanol/methylene chloride to give pure amine 0.778g (64%).
IH NMR (DMSO) d 3.32 (s, 3H), 4.30 (s, IH), 6.64 (d, d, IH), 6.76 (d, IH), 7.35 (dt, IH), 7.58-7.74 (m, 3H), 7.88 (m, IH).
EXAMPLE 84
N-[2,3-dihydro- 1 -methyl-2-oxo-5-(2-furanyl)- 1 H- 1 ,4-benzodiazepin-3- yll-3-cvclohexylpropanamide m.p. 168-169°C
CHN: Anal. Calcd. for C23H27N3O3:
C, 70.21 ; H, 6.92; N, 10.68 Observed: C, 70.15; H, 6.67; N, 10.64 EXAMPLE 85
N-[2,3 -dihydro- 1 -methy l-2-oxo-5-(2-furany 1)- 1 H- 1 ,4-benxodiazepin-3- yl1-3-(4-trifluoromethylphenyPpropanamide m.p. 155-157°C
CHN: Anal. Calcd. for C24H20N3O3F3:
C, 63.29; H, 4.432; N, 9.23 Observed: C, 63.22; H, 4.44; N, 9.07
EXAMPLE 86
N-[2,3-dihydro-l-methyl-2-oxo-5-(2-mranyl)-lH-l,4-benzodiazepin-3- yn-3-(2.4-dichlorophenyl propanamide m.p. 132-133°C
CHN: Anal. Calcd. for C23H19N3O3CI2
C, 60.54; H, 4.20; N, 9.21 Found: C, 60.62; H, 4.07; N, 9.07
EXAMPLE 87
N-[2,3-dihydro- 1 -methy l-2-oxo-5-(3-furanyl)- IH- 1 ,4-benzodiazepin-3- yl 1 - 3-c vcl ohex ylpropanamide m.p. 199-200°C
!H NMR (CDC13) d: 0.9-1.8 (brm, 3H), 2.38 (t, 2H), 3.42 (S, 3H), 5.55
(brd, IH), 6.90 (S, IH), 7.2-7.77 (m, 7H)
EXAMPLE 88
N-[2,3-Dihydro-l-methyl-2-oxo-5-(3-furanyl)-lH-l,4-benzodiazepin-3- yll-3-(4-trifluoromethylphenyl )propanamide m.p. 213-214°C
Ϊ H NMR (CDCI3) d: 2.71 (dt, 2H), 3.05 (t, 2H), 3.42 (S, 3H), 5.72 (d,
IH), 6.82 (brS, IH), 7.2-7.7 (m, 1 1H) EXAMPLE 89
N-[2,3-Dihydro-l-methyl-2-oxo-5-[2'-(4,4-dimethyl-2-oxazolinyl)- pheny 1] -lH-1.4-benzodiazepin-3-yl1-3-(2.4-dichlorophenyI)- propanamide
The subject compound was prepared substantially as described in Example 81. m.p. 194-195°C CHN: Anal. Calcd. for C30H28N4O3CI2
C, 63.95; H, 5.01 ; N, 9.94 Found: C, 63.70; H, 5.01; N, 9.96
EXAMPLE 90
N-[2,3,4,5-Tetrahydro-l -methyl-2-oxo-5-isopropyl-lH-l ,4-benzo- diazepin-3yll-3-cvclohexylpropanamide
Figure imgf000122_0001
A solution of N-[2,3-dihydro-l-methyl-2-oxo-5-isopropyl- lH-l,4-benzodiazepin-3-yl]-3-cyclohexyIproρanamide (50 mg) in methanol (10 mL), containing 10% Pd/C (50 mg) was stirred under 1 atmosphere of hydrogen for 18 hours. Filtration of the reaction, concentration and crystallization ffrom diethyl ether gave 21 mg N- [2,3,4,5-tetrahydro-l -methyl-2-oxo-5-isopropyl-lH-l ,4-benzodiazepin- 3-yl]-3-cyclohexylpropanamide. CHN: Anal. Calcd. for C22H33N3O2
C, 71.12; H, 8.95; N, 1 1.31 Observed: C, 70.98; H, 8.97; N, 11.15 m.p. 114-115°C
EXAMPLE 91
N-[2,3-dmydro-l-methyl-2-oxo-5-methyl-lH-l ,4-benzodiazepin-3-yl]- 3-(2.4-dichlorophenyl)propanamide
Step A:
Figure imgf000123_0001
To CBZ-benzodiazepine (250 mg, 0.776 mmol) in toluene (25 mL) at reflux was added dropwise a solution of DMF dimethylacetal (1.09 mL) in toluene (10 mL). The reaction was refluxed for 5 hours, cooled and concentrated to an oil. The oil was triturated with ether to give a white solid (124 mg).
IH NMR (CDCI3) d: 2.50 (s, 3H), 3.42 (s, 3H), 5.12-5.20 (m, 3H), 6.62 (d, IH), 7.25-6.4 (m, 7H), 7.5-7.6 (m, 2H). Step B:
Figure imgf000124_0001
The CBZ-amine-N -methyl amide (190 mg) was treated with 30% HBr/AcOH (0.8 mL) for 1 hour at room temperature. The reaction mixture was poured into ether (10 mL) at 0°C and the solid filtered. Solid dissolved in 10% Aq. NaOH (5 mL) and CH2CI2 (10 mL) and organic layer separated, dried (Na2Sθ4), filtered and concentrated to an oil (172 mg, 110%). iH NMR (CDCI3) d: 2.42 (s, 3H), 3.05 (brs, 2H), 3.40 (s, 3H), 4.40 (s, IH), 7.2-7.6 (m, 4H).
Step C:
Figure imgf000124_0002
N-[2,3-dihydro-l -methyl-2-oxo-5-methyl-lH-l ,4-benzodiazepin-3-yl]-3-
(2,4-dichlorophenyl)propanamide was prepared in a similar manner as described previously in Example 43. m.p. 194-195°C
CHN: Anal. Calcd. for C20H19N3O2CI2
C, 59.42; H, 4.74; N, 10.39 Observed: C, 59.50; H, 4.74; N, 10.44 iH NMR (CDCI3) d: 2.49 (brs, 3H), 2.65 (dt, 2H), 3.05 (t, 2H), 3.42 (s, 3H), 5.35 (d, IH), 71 -7.6 (m, 8H).
EXAMPLE 92
N-[2,3-Dihydro-l -methyl-2-oxo-[4,5-a]-( 1-oxo- 1 , 3-dihydro-2H- isoindole)-lH-l ,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)- propanamide
Figure imgf000125_0001
To a solution of N-[2,3-dihydro-l-methyI-2-oxo-5-[2'-(4,4- dimethyI-2-oxazolinyl)phenyl]-lH-l ,4-benzodiazepin-3-yl]-3-(2,4- dichlorophenyl)propanamide (100 mg, 0.178 mmol) in methylene chloride was slowly added methyl trifluoromethanesulfonate (22 mL, 0.198 mmol). After stirring 5 minutes, sodium borohydride (7.6 mg, 0.20 mmol) in asolute ethanol (0.5 mL) was added and reaction stirred 30 min. the product was extracted into ethyl acetate and purified by column chromatography on silica (60% ethyl acetate/hexane) to give 30 mg N-[2,3-dihydro-l-methyl-2-oxo-[4,5-a]-(l -oxo-1 , 3-dihydro-2H- isoindole)-l H- 1 ,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)- propanamide. iH NMR (CDC13) d: 2.70 (m, 2H), 3.12 (t, 2H), 3.55 (s, 3H), 5.68 (s, IH), 5.90 (d, IH), 6.85 (dd, IH), 7.05 (brd, IH), 7.1 -7.5 (m, 9H), 7.85 (d, IH). MS M+l -494.
EXAMPLE 93
Figure imgf000126_0001
3R-(+)-3-(Phenylthio)-N-[2,3-dihydro-l -methyl-2-oxo-5-phenyl-lH-
1.4-benzodiazepin-3-yllpropanamide
To a stirred solution of 3-bromopropionic acid (l.Og, 6.5mmol) in DMF (20 mL) was added K2CO3 (1.8 g, 13 mmol) and thiophenol (0.72 g, 6.5 mmol). This was heated to 50°C for lh. The mixture was then diluted with 200 mL H2O and extracted with 2 x 100 mL EtOAc. The combined organics were washed with 100 mL H2O and dried with Na2S04. This was evaporated to give 1.52g of a colorless oil, 1.18g corrected for residual DMF by NMR. The above oil was taken up in 30 mL DMF and l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.45g, 12.8mmol) and 1-hydroxybenztriazole hydrate ( 1.73g, 12.8mmol) were added. This was stirred for 5 min at rt. 3-(R)- Amino- 1,3-dihydro-l - methyl-5-phenyl-2H-l ,4-benzodiazepin-2-one (0.66g, 2.6mmol) was then added and the reaction was stirred at rt overnight. The reaction was diluted with 200 mL H2θ and extracted with 2xl50mL EtOAc. The combined organics were washed with lxlOOmL H2θ, dried with Na2Sθ4 and evaporated. The residue was chromatographed over silica eluting with 2% MeOH:CHCl3. Collected pure fractions, evaporated. Evaporated from diethyl ether to give 770mg of a white foam. Anal. Calcd for C25H23N3θ2S-0.05Hexane:
C, 70.04; H, 5.51; N, 9.69. Found: C 69.91 , H 5.40, N 9.78.
EXAMPLE 94
Figure imgf000127_0001
3R-(+)-5-(Methy lthio)-N-[2,3-dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H-
1.4-benzodiazepin-3-yl1propanamide
To an aqueous solution of K2CO3 (0.76g, 5.5mmol) was added 5-bromopentanoic acid and sodium thiomethoxide. This was stirred at rt overnight. The reaction was diluted with 50 mL H2O and acidified to pH=0 with 6N HCl. Extracted with 2 x 50 mL EtOAc. Dried with Na2Sθ4, evaporated to give 0.55g of a yellow oil. The above oil was taken up in 10 mL DMF and l -(3- dimethyl-aminopropyl)-3-ethylcarbodiimide hydrochloride (1.30g, 6.8mmol) and 1-hydroxybenztriazole hydrate (0.92g, 6.8mmol) were added. 3-(R)-Amino-l ,3-dihydro-l -methyl-5-phenyl-2H-l ,4- benzodaizepin-2-one (0.85g, 3.4 mmol) was then added and the reaction was stirred overnight at rt. The reaction was diluted with 100 mL H2θ and extracted with 2 x 50 mL EtOAc. Combined organics were dried with brine and Na2Sθ4, and evaporated to give yellow oil. The residue was chromatographed over silica eluting with 50:50 EtOAc:Hex to 100% EtOAc. Pure fractions were collected to give 1.33g of a colorless oil, 0.4g of which was chroma-tographed over silica eluting with 2% MeOH:CH2Cl2- Pure fractions were collected, and evaporated from ethyl etheπhexane to give a white powder mp. 61-65°C. Anal. Calcd for C22H25N3O2SO.35H2O: C, 65.76; H, 6.45; N, 10.46. Found: C, 65.81 ; H, 6.21; N, 10.57.
EXAMPLE 95
Figure imgf000128_0001
N-cyano-N'-cyclohexylmethyl-N"-(l ,3-dihydro-l -methy l-2-oxo-5- phenyl-2H- 1.4-benzodiazepin-3-vPguanidine
A solution of 3-(R)-amino-l ,3-dihydro-l -methyl-5-phenyl- 2H-l ,4-benzodiazepin-2-one (lg, 3.7 mmole) in acetonitrile (20 mL) was treated with diphenylcyanocarbonimidate (0.9 g, 3.7 mmole) and stined at room temperature for thirty minutes. Cyclohexylmethyl- amine (0.84 g, 7.4 mmole) was then added and the reaction stirred at room temperature for two hours. The reaction was poured into 100 mL of 0.1 N HCl and extracted with 3 x 100 mL portions of ethyl acetate. The organic layers were combined and washed once with saturated sodium bicarbonate (50 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated at reduced pressure. The residue was chromatographed on silica gel eluting with 50% ethyl acetate/hexane to give 0.875 g of the product. The analytical sample was crystallized from ethyl acetate, m.p. 158-161 °C. Anal. Calcd. for C25H28N60:
C, 70.07; H, 6.59; N, 19.61. Found: C, 70.05; H, 6.59; N, 19.64%.
EXAMPLE 96
Figure imgf000129_0001
N-( 1 ,3-Dihydro- 1 -methy l-2-oxo-5-pheny 1-2H- 1 ,4-benzodiazepin-3-yl)- 4-(4-chlorobenzylV4-piperidinecarboxamide dihvdrochloride
Step A: Preparation of N-tert-butyloxycarbonyl-4-(4-chloro- benzyl)-4-piperidinecarboxylic acid
A solution of N-Boc-ethylisonipecotate (51.4 g, 200 mmole) in THF (1L) at -60° C was treated with a solution of lithium bistrimethylsilyl amide (220 mL of a 1 N solution in THF, 220 mmole). After stirring at -60°C for 5 minutes, a solution of 4-chlorobenzyl chloride (33.8 g, 210 mmole) in THF (200 mL) was added and the reaction allowed to warm to room temperature. Most of the THF (about 800 mL) was removed by evaporation at reduced pressure. The remainder was poured into 1 L of 1 N HCl and extracted with two 800 mL portions of ethyl acetate. The organic layers were combined and washed once with saturated sodium bicarbonate (500 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated at reduced pressure. The residue was chromatographed on silica gel eluting with 10%-20% ethyl acetate/hexane to give the product ester which was used directly. The material thus obtained was dissolved in THF (100 mL) and IPA (100 mL) and treated with 350 mL of 10 N NaOH. The mixture was heated to reflux for 30 hours. The reaction was cooled to room temperature and poured over a mixture of crushed ice (2 L), 6 N HCl (500 mL) and saturated potassium hydrogen sulfate (1 L). The mixture was extracted with two 1 L portions of ethyl acetate. The organic layers were combined and dried over anhydrous magnesium sulfate, filtered, and concentrated at reduced pressure to give 52 g of the product, m.p. 179-180°C, iH NMR CDC13 d 7.26 (d, J = 8 Hz, 2 H), 7.03 (d, J = 8 Hz, 2 H), 3.98 (m, 2H), 3.0-2.8 (m, 2H), 2.84 (s, 2H), 2.10-2.00 ( m, 2H), 1.55- 1.40 (m, 2H), 1.45 (s, 9H)
Step B: Preparation of N-( 1 ,3-dihydro- 1 -methyl-2-oxo-5-phenyl- 2H- 1 ,4-benzodiazepin-3-yl)-4-(4-chlorobenzyl)-4-piper- idinecarboxamide dihydrochloride
A mixture consisting of N-tert-butyloxycarbonyl-4-(4- chlorobenzyl)-4-piperidinecarboxylic acid (1.48 g, 4.18 mmole), 3-(R)- amino- 1 ,3-dihydro- 1 -methyl-5-pheny 1-2H- 1 ,4-benzodiazepin-2-one ( 1 g, 3.7 mmole), hydroxybenzotriazole (1.17 g, 8.66 mmole), l-(3-dim- ethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.49 g, 7.70 mmole), diisopropylethyl amine (0.53 g, 4.13 mmole), and DMF (10 mL) was stirred at room temperature for 18 hours. The reaction was poured into 1 N HCl and extracted with ethyl acetate (4 X 50 mL). The organic layers were combined and washed once with saturated sodium bicarbonate (50 mL), once with saturated sodium chloride (50 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated at reduced pressure. The residue was chromatographed on silica gel eluting with 25%-50% ethyl acetate/hexane to give 2.34 g of the product amide which was used directly. The material thus obtained was dissolved in ethyl acetate (50 mL) and HCl (g) was bubbled into the reaction for 5 minutes. The reaction was concentrated at reduced pressure and the residue recrystallized from ethyl acetate to give 1.13 g of the product as a pale yellow solid, m.p. 190 - 195°C.
Anal. Calcd. for C29H29ClN4θ2«2 HCl: C, 60.68; H, 5.44; N, 9.76. Found: C, 60.47; H, 5.5; N, 9.42%.
Utilizing the procedures substantially as desribed above except substituting N-Boc-ethylnipecotate for N-Boc-ethyl isonipecotate there were obtained the following compounds
EXAMPLE 97
Figure imgf000131_0001
N-(l ,3-dihydro-l-methyl-2-oxo-5-phenyl-2H-l,4-benzodiazepin-3-yl)- 3-(4-chlorobenzyl)-3-piperidinecarboxamide hydrochloride A + B isomers Isomer A m.p. 205 - 210°C.
Anal. Calcd. for C29H28C1N4O2-HC1-0.5 CH3CH2OH-0.8 H2θ:
C, 62.67; H, 6.07; N, 9.75. Found: C, 62.69; H, 5.94; N, 9.42%.
Isomer B m.p. 200 - 205°C.
Anal. Calcd. for C_9H28C.N4O2-HCl.-0.1 CH3CH20COCH3-1.6 H2O:
C, 61.39; H, 5.96; N, 9.74. Found: C, 61.39; H, 5.66; N, 9.56%.
EXAMPLE 98
Figure imgf000132_0001
Figure imgf000132_0002
(+)-3-Cyclohexyl-N-[2,3-dihydro- 1 -methyl-2-oxo-5-pheny 1- 1 H- 1 ,4- benzodiazepin-3-yll-N-(ethoxycarbonylmethyl)propanamide
3-(R)- Amino- 1 ,3-dihydro- 1 -methy 1-5-pheny 1-2H- 1 ,4- benzodiazepin-2-one (5.0 g, 18.8 mmol) in acetonitrile (100 mL) was mixed with ethyl bromoacetate (2.1 mL, 18.8 mmol) and sodium hydrogen carbonate (4.0 g) was suspended in the mixture. The mixture was stirred and heated at reflux for 2 h. After that time, the reaction was cooled to room temperature, diluted with 150 mL water, and extracted with ethyl acetate (3 x 100 mL). The organic layers were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed in vacuo. The resulting oil was chromatographed over silica in 3: 1 ethyl acetate:hexane, yielding the mono-alkylated product (2.58 g, 39%) as well as the starting 1 ,4-benzodiazepin-2-one and bis-alkylated material. To a solution of 3-cyclohexylpropionic acid (1.0 g, 6.40 mmol) in methylene chloride (30 mL) was added oxalyl chloride (0.56 mL, 6.40 mmol) and catalytic (N,N)-dimethyl formamide ( 2 drops). After 0.5 h, a solution of the acetate (2.25 g, 6.40 mmol) in methylene chloride (10 mL) was added and the reaction was stirred for 0.25 h. The reaction was then diluted with methylene chloride (150 mL) and saturated aqueous sodium hydrogen carbonate (150 mL ) was added. The aqueous portion was extracted again with methylene chloride (2 x 100 mL) and the organics were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed in vacuo. The resulting oil was chromatographed over silica with 1 :1 ethyl acetate :hexane, yielding a foam that was crystallized with ether, giving 2.0 g (64%) of the product, m.p. 120-122°C, [α]D + 0.63° (c=0.79; MeOH). Anal. Calcd. for C29H35N3O4:
C, 71.14; H, 7.21; N, 8.58. Found: C, 71.13; H, 7.13; N, 8.75%.
The following compound was prepared in a manner substantially as desribed above except substituting ethyl bromobutyrate for ethyl bromoacetate. EXAMPLE 99
Figure imgf000134_0001
3-Cyclohexy 1-N-f 2,3-dihydro- 1 -methy l-2-oxo-5-phenyl- 1 H- 1 ,4- ber_odiazepin-3-yll-N-(ethoxycarbonylpropyDpropanamide m.p. 103-105°C, [α]D 0.00°; c=0.85; MeOH. Anal. Calcd. for C3iH39N3O4.-O.4O mol H2θ:
C, 70.94; H, 7.64; N, 8.01. Found: C, 70.91; H, 7.44; N, 8.12%.
EXAMPLE 100
Figure imgf000135_0001
N-[2,3-Dihydro-l-methyI-2-oxo-5-phenyl-lH-l,4-benzodiazepin-3-yl]-
N-r2-(2-methoxyethoxytethyl1hexanamide
3-(R)-Amino- 1 ,3-dihydro- 1 -methyl-5-phenyl-2H- 1 ,4- benzodiazepin-2-one (1.33 g, 5.0 mmol) in N,N-dimethyl formamide (30 mL ) was mixed with l-bromo-2-(2-methoxyetho y)ethane (1.35 mL , 5.0 mmol) and triethylamine (1.0 mL ). The mixture was stirred and heated at reflux for 4 h. After that time, the reaction was cooled to room temperature, diluted with 150 mL water, and extracted with ethyl acetate (3 x 100 mL ). The organic layers were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed in vacuo. The resulting oil was chromatographed over silica in 1 : 1 ethyl acetate :hexane, yielding the mono-alkylated product (1.2 g, 65%) as well as the starting 1 ,4-benzodiazepin-2-one and bis-alkylated material. To a solution of the mono-alkylated material (1.2 g, 3.27 mmol) in methylene chloride (20 mL) was added hexanoyl chloride (0.96 mL, 3.27 mmol) and the reaction was stured for 0.25 h. The reaction was then diluted with methylene chloride (150 mL) and saturated aqueous sodium hydrogen carbonate (150 mL) was added. The aqueous portion was extracted again with methylene chloride (2 x 100 mL) and the organics were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed in vacuo. The resulting oil was chromatographed over silica with 1 : 1 ethyl acetate: hexane, yielding an oil, giving 580 mg (38%) of the product. [α]D 0.00°; c=0.27; MeOH. Anal. Calcd. for C27H35N3O4.-O.8O mol H2O:
C, 67.56; H, 7.69; N, 8.75. Found: C, 67.56; H, 7.39; N, 8.85%.
EXAMPLE 101
Figure imgf000137_0001
(+)-N-[2,3-Dihydro- 1 -methyl-2-oxo-5-pheny 1- 1 H- 1 ,4-benzodiazepin-3- yll-N-(5-hvdroxypentvπhexanamide
3-(R)-Amino- 1 ,3-dihydro- 1 -methy l-5-phenyl-2H- 1 ,4- benzodiazepin-2-one (1.33 g, 5.0 mmol) in acetonitrile (40 mL ) was mixed with 5-chloropentan-l-ol (0.61 g, 5.0 mmol) and sodium hydrogen carbonate (2.0 g) was suspended in the mixture. The mixture was stined and heated at reflux for 12 h. After that time, the reaction was cooled to room temperature, diluted with 100 mL water, and extracted with ethyl acetate (3 x 75 mL ). The organic layers were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed hi vacuo. The resulting oil was chromatographed over silica in 1 :49 methanolxhloroform yielding the mono-alkylated product (1.1 g, 62%) as well as the starting 1 ,4-benzodiazepin-2-one and bis-alkylated material. To a solution of the monoalkylated material (0.50 g, 1.42 mmol) in methylene chloride (30 mL ) was added hexanoyl chloride (0.20 mL, 1.42 mmol) and the reaction was stirred for 0.25 h. The reaction was then diluted with methylene chloride (100 mL) and saturated aqueous sodium hydrogen carbonate (100 mL ) was added. The aqueous portion was extracted with methylene chloride (2x75 mL ) and the organics were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed jn vacuo. The resulting oil was chromatographed over silica with 1 : 1 ethyl acetate:hexane, yielding a foam, giving 360 mg (64%) of the product, foam , [α]d + 8.36° (c=0.61, MeOH). Anal. Calcd. for C27H35N3O2.-O.25 mol H2O:
C, 71.42; H, 7.88; N, 9.25. Found: C, 71.47; H, 7.89; N, 9.12%.
EXAMPLE 102
Figure imgf000139_0001
(+)-N-[2,3-Dihydro-l -methyl-2-oxo-5 -phenyl- 1 H- 1 ,4-benzodiazepin-3- yll-N-(ethoxycarbonylpentyl hexanamide
3-(R)- Amino- 1 ,3-dihydro-l -methy l-5-phenyl-2H- 1,4- benzodiazepin-2-one (1.33 g, 5.0 mmol) in acetonitrile (40 mL ) was mixed with ethyl-6-bromohexanoate (0.89 mL , 5.0 mmol) and sodium hydrogen carbonate (2.0 g) was suspended in the mixture. The mixture was stined and heated at reflux for 10 h. After that time, the reaction was cooled to room temperature, diluted with 100 ml . water, and extracted with ethyl acetate (3x75 mL ). The organic layers were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed in vacuo. The resulting oil was chromatographed in 1 :49 methano chloroform, yielding the mono-alkylated product (0.56 g, 28%) as well as the starting 1 ,4-benzodiazepin-2-one and bis- alkylated material. To a solution of the mono-alkylated material (0.56 g, 1.37 mmol) in methylene chloride (20 mL) was added hexanoyl chloride (0.19 mL, 1.37 mmol) and the reaction was stined for 0.25 h. The reaction was then diluted with methylene chloride (100 mL) and saturated aqueous sodium hydrogen carbonate (100 mL) was added. The aqueous portion was extracted again with methylene chloride (2x75 mL) and the organics were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed in vacuo. The resulting oil was chromatographed over silica with 1 :1 ethyl acetate: hexane, yielding a foam, giving 0.40 g (58%) of the product, m.p. 59-65°C, l ]d (+)52.7° (c=0.48,MeOH). Anal. Calcd. for C30H39N3O4.-O.2O mol CH2CI2:
C, 69.4; H, 7.6; N, 8.04. Found: C, 69.44; H, 7.68; N, 7.71%.
The following compound was prepared in a manner substantially as described above except substituting ethyl bromoacetate for ethyl 6-bromohexanoate.
EXAMPLE 103
Figure imgf000141_0001
(+)-N-[2,3-Dihydro- 1 -methyl-2-oxo-5-pheny 1- 1 H- 1 ,4-benzodiazeρin-3 ■ yl1-N-(emoxycarbonylmethvDhexanamide foam, [α]d + 2.04° (c=0.98; MeOH). Anal. Calcd. for C26H31N3O4:
C, 69.47; H, 6.95; N, 9.35. Found: C, 69.41; H, 7.03; N, 9.26%.
EXAMPLE 104
Figure imgf000141_0002
(+)-3-Cyclohexyl-N-[2,3-dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1 ,4- benzodiazepin-3-yl1-N-(hydroxymethyl)propanamide
(+)-3-Cyclohexyl-N-[2,3-dihydro- 1 -methyl-2-oxo-5- phenyl-lH-l ,4-benzodiazepin-3-yl]propanamide (2.0 g, 5.0 mmol) was dissolved in tetrahydrofuran (30 mL), cooled to 0°C and methyl magnesium chloride (3M, 2.0 mL) was added. After 0.25 h, paraformadehyde (0.15 g, 10 mmol) was added, and the mixture was allowed to warm to room temperature. The reaction was then diluted with ethyl acetate (150 mL) and saturated aqueous sodium hydrogen carbonate (150 mL) was added. The aqueous portion was extracted again with ethyl acetate (2 x 100 mL) and the organics were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed in vacuo. The resulting oil was chromatographed over silica with 1:1 ethyl acetate:hexane, yielding a foam (0.80 g, 37%). foam, [α]d + 124° (c=0.69, MeOH). Anal. Calcd. for C26H31N3O3:
C, 72.03; H, 7.21; N, 9.69. Found: C, 71.66; H, 7.08; N, 9.78%.
The following compound was prepared 111 a manner substantially as described above starting from (+)-N-[2,3-dihydro-l- methyl-2-oxo-5-phenyl-lH-l,4-benzodiazepin-3-yl]hexanamide.
EXAMPLE 105
Figure imgf000142_0001
(+)-N-[2,3-D ydro-l -me yl-2-oxo-5-phenyl-lH-l,4-benzodiazepin-3- yll-N-(hvdroxymethyl)hexanamide m.p. 154-156°C, [α]d + 190.8° (c=0.24 , MeOH). Anal. Calcd. for C23H27N3O3O.3O mol H2O: C, 69.26; H, 6.97; N, 10.53. Found: C, 69.29; H, 6.81; N, 10.6%.
EXAMPLE 106
Figure imgf000143_0001
(+)-3-Cyclohexyl-N-[2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH-l ,4- benzodiazepin-3-yll-N-(tetrazolylmethyl)propanamide
(+)-3-Cyclohexyl-N-[2,3-dihydro- 1 -methyl-2-oxo-5- phenyl-lH-l ,4-benzodiazepin-3-yl]-N-(hydroxymethyl)propanamide (0.67 g, 1.56 mmol) was dissolved in methylene chloride(100 mL), along with tetrazole (0.33 g, 4.7 mmol), and then N,N-diisopropyl- dibenzyl-phosphoramidite (1.07 g, 3.1 mmol). After 2 h, the mixture was diluted with methylene choride (150 mL), and extracted with saturated aqueous sodium hydrogen carbonate (3 x 100 mL). The organic layers were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed in vacuo. The resulting oil was chromatographed twice over silica with 1 :1 ethyl acetate:hexane, yielding two constitutional isomers, a (65 mg, 9%) and b (56 mg, 7.5%).
Isomer A: m.p. 96-98°C, [α]d +188.9° (c=0.19, MeOH).
Anal. Calcd. for C27H31N7O2O.3O mol TFA:
C, 63.78; H, 6.07; N, 18.86. Found: C, 63.7; H, 6.12; N, 18.76%. Isomer B: m.p. 92-95°C, [α]d +81.3° (c=0.31, MeOH).
Anal. Calcd. for C27H31N7O2OJ5 mol TFA:
C, 63.31; H, 6.01; N, 18.66. Found: C, 63.35; H, 6.02; N, 18.74%.
EXAMPLE 107
Figure imgf000144_0001
3R-(+)-3-(Benzyloxycarbonylamino)-2,3-dihydro-l-methyl-2-oxo-5- phenyl- 1 H- 1.4-benzodiazepine
To a stirring solution of 3-(R)-amino-l,3-dihydro-l- methyl-5-phenyl-2H-l,4-benzodiazepin-2-one (2.0 g, 7.5 mmol) in methylene chloride (45 mL ) at 0°C was added benzyl chloroformate (1.2 mL, 8.3 mmol) and the reaction was allowed to warm to room temperature. The reaction mixture was diluted with methylene chloride (150 mL ), and extracted with saturated aqueous sodium hydrogen carbonate (150 mL ). The aqueous portion was extracted with methylene chloride (2 x 100 mL ) and the organics were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed in vacuo. The resulting oil was chromatographed over silica with 1:1 ethyl acetate :hexane, yielding a white foam (3.0 g, 99.7%) [α]d +57.5° (c-1.17; MeOH). Anal. Calcd. for C24H20N3O3-- 0.70 mol H2O -0.15 mol CHCI3:
C, 67.62; H, 5.06; N, 9.8. Found: C, 67.6; H, 5.02; N, 9.75%. The following compounds were prepared substantially as described in Example 81.
EXAMPLE 108
N-[2,3-Dihydro-l-methyl-2-oxo-5-ethyI-lH-l ,4-benzodiazeρin-3-yl]-3-
(2.4-dichlorophenvPpropanamide m.p. 156-158°C.
CHN: Anal. Calcd. for C2lH2lCl2N3θ2-0.5 H2O:
C, 59.02; H, 5.19; N, 9.83. Found: C, 58.99; H, 4.89; N, 9.88.
EXAMPLE 109
N-[2,3-Dihydro-l-methyl-2-oxo-5-t-butyl-lH-l ,4-benzodiazepin-3-yl]-
3-(2.4-dichlorophenvDpropanamide m.p. 170-171 °C.
CHN: Anal. Calcd. for C23H25C12N3O2-0.7 H2O:
C, 60.18; H, 5.80; N, 9.16. Found: C, 60.17; H, 5.30; N, 9.30.
EXAMPLE 110
N-[2,3-Dihydro- l -methyl-2-oxo[4'-(4,4-dimethyl-2-oxazolinyl)phenyl]- lH-1.4-benzodiazepin-3-yl1-3-(2.4-dichlorophenyPpropanamide m.p. 188-190°C.
CHN: Anal. Calcd. for C30H28N4O3CI2:
C, 63.95; H, 5.01; N, 9.94. Found: C, 63.96; H, 5.02; N, 10.08.
EXAMPLE 111
N-[2,3-Di ydro- 1 -methyl-2-oxo-5-(4-methoxyphenyl)- IH- 1 ,4- benzodiazepin-3-yll-3-(2.4-dichlorophenvDpropanamide m.p. 188-189°C.
CHN: Anal. Calcd. for C26H23C12N3O3-0.45 H2θ:
C, 62.91 ; H, 4.67; N, 8.47. Found: C, 61.89; H, 4.78; N, 8.33.
EXAMPLE 112
(-r 3,5-Dichloro-N-[3R-2,3-dihydro-2-oxo-5-phenyl-l -(2,2,2- trifluoroethylV 1 H-benzofel \ 1 ,4ldiazepin-3- yllbenzamide.
Figure imgf000146_0001
Step A: Preparation of 2,3-dihydro-2-oxo-5-phenyl-l -(2,2,2- trifluoroethyl)- 1 H-benzo[e] [ 1 ,4]diazepine.
A solution of 5-phenyl-l,4-benzodiazepine-2-one (J. Org. Chem., 1962, 27, 3788)(50 g, 0.211 mole) in DMF (100 mL) was treated with cesium carbonate (103.5 g, 0.317 mole) and trifluoroethyl iodide (109.7 g, 0.525 mole). The mixture was stirred at 50°C overnight. The reaction mixture was then poured into water (2 L) and extracted with ethyl acetate (3 X 1 L). The combined ethyl acetate fractions were dried over anhydrous magnesium sulfate, filtered and concentrated at reduced pressure. The residue was crystallized from ethyl ether to give 45 g (68 %) of the product. MP = 130 - 131°C; iH NMR (CDCI3, 300 MHz) d 7.65-7.60 (m, 2H), 7.60-7.45 (m, 5H), 7.40-7.20(m, 2H), 5.25 (dq, J = 14, 8.6 Hz, IH), 4.82(d, J = 10.5 Hz, IH), 4.15 (app sextet, J = 8.6 Hz, IH), 3.81 (d, J = 10.5 Hz, IH) Step B: Preparation of 3-Azido-5-phenyl-l -(2,2,2-trifluoroethyl)- 1 H-benzo[e] f 1 ,4]diazepine.
To a stirring solution of 5-phenyl-l-(2,2,2-trifluoroethyl)- lH-benzo[e][l,4]diazepine (70 g,0.22 mol) in THF (1500 mL) cooled to -70°C was added potassium tert-butoxide(l .l eq, 0.24 mol, 240 mL of a 1 N solution in THF) dropwise over 15 min. A solution of 2,4,6- triisopropylbenzenesulfonylazide (74.8 g, 0.24 mol) in THF (250 ml) was added over 5 min. This was stirred for 10 minutes and acetic acid (40 mL, 0.63 mol) was added and the reaction allowed to warm to ambient temperature. The reaction was poured into satd. NaHC03 (1500 mL) and ethyl acetate (1L). The phases were separated and the aqueous phase was extracted with ethyl acetate(500 mL). The combined organic layers were washed with water (500 mL) then brine (300mL). The organic layers were dried with Na2S04 and evaporated to a brown foam. This was triturated with ethyl ether to give 65 g of a white powder. The filtrate was concentrated and chromatographed over silica gel eluting with 30% ethyl acetate/hexane to give another 8.9 g. The combined yield was 74 g( 93%). MP = 159 - 160°C; iH NMR (CDC13, 300 MHz) d 7.70-7.26 (m,9H), 5.28-5.12 (m,lH), 4.63 (s,lH), 4.35-4.10 (m,lH).
Step C: Preparation of racemic 3-Amino-5-phenyl-l -(2,2,2- trifluoroethyl)- 1 H-benzo[e] [ 1 ,4]diazepine.
To a stirring solution of 3-Azido-2-oxo-5-phenyl-l -(2,2,2- trifluoroethyl)-2,3-dihydro- 1 H-benzo[e] [ 1 ,4]diazepine (83.4mmol,30g) in 300mL ethanol and 150mL THF was added 10%Pd/C (10 wt%, 3.0g). Hydrogen gas was bubbled through the solution for 8h. The reaction was filtered and evaporated under reduced pressure. The residue was crystallized from ethyl ether to give 20.0g of white crystals. Another 4g was recovered from evaporation and recrystallization of the filtrates. Combined yeild, 86.7%. MP = 141 - 143°C; iH NMR (CDC13.300 MHz) d 1.10-1.26 (m,9H), 5.28-5.12 (m,lH),
4.57 (s,lH), 4.35-4.10 (m,lH).
Step D: Preparation of 2-Amino-N-[2-oxo-5-phenyl- 1 -(2,2,2- trifluoroethyl)-2,3 -dihydro- 1 H-benzo[e] [ 1 ,4]diazeρin- 3-yl]-3-phenylpropionamide
To a stirring solution of 3-Amino-2-oxo-5-phenyl-l -(2,2,2- trifluoroethyl)-2,3-dihydro-lH-benzo[e][l,4]diazepine (92.2 mmol, 30.74g) in DMF (300mL) was added N-Benzyloxy-D-Phenylalanine (92.2 mmol, 27.6g), l-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.12mol,22.95g) and 1-hydroxybenztriazole hydrate (46.1mmol,6.23g). This was stined at room temperature for 2h. The reaction was then diluted with 1L of 10% KHSO4 and extracted with ethyl acetate (2x600 mL). The organic layers were combined and washed with saturated sodium hydrogen carbonate (600mL). They were dried with brine and sodium sulfate and evaporated under reduced pressure. 66.58g of an orange foam, which contained ethyl acetate by NMR. NMR 1H (CDC13) d 7.75-7.18 (m, 20H), 5.62-5.55 (m,lH), 5.48-5.00 (m, 4H), 4.72-4.60 (m, IH), 4.25-4.05 (m,lH) 3.32-3.05 (m, 2H). This material was carried on without further purification.
To a stirring solution of 2-(N-Benzyloxyamino)-N-[2-oxo- 5-pheny 1- 1 -(2,2,2-trifluoroethyl)-2,3-dihydro- 1 H-benzo[e] [ 1 ,4]diazepin- 3-yl]-3-phenyl propionamide in 1L ethanol was added 10% Pd/C (15 wt%) and hydrogen was bubbled through the reaction for 2h and then left stirring under 1 atm. hydrogen overnight. Hydrogen was bubbled through the reaction for an additional three hours the following morning. The reaction was then filtered, the catalyst was rinsed with 1L methylene chloride and evaporated under reduced pressure. The resulting solid was dried under vacuum overnight to give 44.46g of a white solid. This was chomatographed over silica, eluting with 1 % MeOH.'EtOAc. The pure upper Rf fractions were collected and evaporated under reduced pressure. The mixed fractions were collected, evaporated and rechromatographed. The pure fractions were collected and combined with the above pure fractions to get a combined yield of 18.1 lg, 83.5% of the upper Rf diastereomer. *H NMR (CDCl3,300 MHz) d 8.94 (d, J=8.6Hz, IH), 7.65-7.10 (m, 9H), 5.64 (d, J=8.6 Hz, IH), 5.28-5.12 (m, IH), 4.57 (s, IH), 4.35-4.10 (m, IH) 3.71 (dd, J=9.8 and 3.9 Hz, IH), 3.34 ( dd, J=13.9 and 3.9 Hz,lH), 2.79 (dd, J-13.9 and 10.0 Hz, IH). The Absolute stereochemistry at C-3 of the benzodiazepine ring was determined to be (R) by X-Ray analysis.
The lower Rf material conesponding to C-3(S) was isolated as well.
Step E: Preparation of 3(R)-(+)-3-Amino-5-phenyl- 1 -(2,2,2- trifluoroethyl)-l H-benzo[e] [1 ,4] diazepine.
To a stirring solution of 2-Amino-N-[2-oxo-5-phenyl-l- (2,2,2-trifluoroethyl)-2,3-dihydro- 1 H-benzo[e][ 1 ,4]diazepin-3-yl]-3- phenylpropionamide (13.6 g, 28.3 mmol) in methylene chloride (136 mL) was added phenyl isothiocynate (3.87 mL, 34.0 mmol). This was stined overnight at ambient temperature. The reaction was then cooled in ice, trifluoroacetic acid (2.73 mL, 0.283 mol) added and the reaction allowed to warm to ambient temperature. After stirring at ambient temperature for 2.5 hours the reaction was evaporated under reduced pressure, chromatographed with 90:10:1 : 1 methylene chloride: methano acetic acid:water. The low Rf spot was collected and evaporated under reduced pressure with no heat. The residue was taken up in 600 mL methylene chloride and washed with 300 mL saturated NaHCθ3 and 300 mL water. The solution was dried over Na2Sθ4 and evaporated under reduced pressure. The residue was crystallized from ethyl acetate :hexanes to give 6.65 g of a white powder . MP = 162 - 164°C;
JH NMR (CDC13,300 MHz) d 7.70-7.26 (m,9H), 5.28-5.12 (m,lH), 4.57 (s,lH), 4.35-4.10 (m,lH). [α]D = +72.9° (c=0.7, MeOH)
The (-)-3S enantiomer was prepared in the same fashion from the Lower Rf product of Step D. MP = 156 - 158°C; iH NMR (CDCl3,300 MHz) d 7.70-7.26 (m,9H), 5.28-5.12 (m,lH), 4.57 (s,lH), 4.35-4.10 (m,lH). [α]D = -71.2° (c=0.66, MeOH)
Step F: Preparation of (+)-3,5-Dichloro-N-[3R-2,3-dihydro-2-oxo-
5- phenyl- 1 -(2,2,2-trifluoroethyl)-l H-benzo[e]l 1 ,4]diazepin-3- yljbenzamide:
To a stirring solution of (+)-3R-3-amino-5-phenyl-l- (2,2,2-trifluoroethyl)-lH-benzo[e][ l,4] diazepine (5.6 g, 16.8 mmol) in DMF (50 mL) was added l-(3-Dimethylaminopropyl-3-ethylcarbodi- imide hydrochloride(4.44 g, 23.0 mmol), and 1-hydroxybenztriazole hydrate (3.11 g, 23.0 mmol) and 3,5-Dichlorobenzoic acid (3.21 g, 16.8 mmol). This was stined at ambient temperature for 2 hours. The reaction was diluted with 500 mL satd. NaHCθ3 and extracted with 2 x 300 mL ethyl acetate. The combined organics were washed with 10% KHSO4 (200 mL) , brine (200 mL), dried over Na2S04, and evaporated to a white foam. This was chromatographed over a 75 x 200 mm silica column eluting with 20% ethyl acetate:hexane. The pure fractions were collected and evaporated under reduced pressure to give 8.5 g of a white foam which was crystallized from 15% ethyl acetate :hexane to give 5.3 g of a white powder . mp=140-143°C, [α]D=+47.9°; U NMR (CDCI3, 300 MHz) d 7.85-7.75 (m, 2 H), 7.70- 7.20 (m, 9 H), 5.78 (d, J=8.1 Hz,l H), 5.30-5.15 (m, IH), 4.30-4.15 (m, IH) Analysis Calcd. for C24H16CI2F3N3O2:
C, 56.93; H, 3.19; N, 8.30; Found: C, 56.81 ; H, 3.17; N, 8.17. The following examples were prepared by a procedure substantially as described for Example 1 , Step F.
EXAMPLE 1 13
(-)-2-(3,4-Dichlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5-phenyl- 1 -(2.2.2-trifluoroethvπ-l H-benzprelll .41diazepin-3-yl1acetamide.
Figure imgf000151_0001
mp=219-221°C; [α]D=-10.8°; iH NMR (CDCl3,300 MHz) d 7.65-7.15 (m, 12H), 5.78 (d, J=8.1 Hz,
IH), 5.25-5.10 (m, IH), 4.25-4.05 (m, IH), 3.56 (s, 2H);
Analysis Calcd. for C25HI 8CI2F3N3O2O.85 H2O:
C, 56.06; H, 3.71 ; N, 7.84. Found: C, 56.03; H, 3.53; N, 7.82.
EXAMPLE 1 14
(-)-2-(3,5-Dichlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5-phenyl- 1 -(2.2.2-trifluoroethylV lH-benzoleir 1 ,4ldiazepin-3-yllacetamide
Figure imgf000152_0001
mp=93-100°C, [α]D = -5.7°;
»H NMR (CDCl3,300 MHz) d 7.65-7.15 (m, 12H), 5.78 (d, J=8.1 Hz, IH), 5.25-5.10 (m, IH), 4.25-4.05 (m, IH), 3.65 (s, 2H); Analysis Calcd. for C25HιgCl2F3N3θ2 : C, 57.71; H, 3.49; N, 8.08; Found : C, 57.41 ; H, 3.48; N, 8.12.
EXAMPLE 1 15
(-)-2-[3,5-Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo- 5-phenyl-l-(2,2,2-trifluoroethyl)-lH-benzo[e][l,4]diazepin-3- yllacetamide
Figure imgf000152_0002
m.p. foam °C, [α]D = -9.7° (c=0.59,MeOH). Anal. Calcd. for C27H18F9N3O2O.75 H20: C, 53.96; H, 3.27; N, 6.99. Found: C, 53.96; H, 3.1 ; N, 6.98%.
EXAMPLE 1 16
(-)-2-(4-Trifluoromethylphenyl)-N-[3R-2,3-dihydro-2-oxo-5- phenyl-1 -(2.2.2-trifluoroethylV lH-benzofeiπ ,41diazepin-3-yllacetamide
Figure imgf000153_0001
m.p. 253-255 °C, [α]D =-9.2° (c=0.25, MeOH).
Anal. Calcd. for C_6Hl 9F6N3θ2-0.05 ethyl etherθ.55 H20:
C, 59.03; H, 3.9; N, 7.88. Found: C, 59.05; H, 3.82; N, 7.78%.
EXAMPLE 1 17
2-(3-Trifluoromethylphenyl)-N-[3R-2,3-dihydro-2-oxo-5-phenyl- l-(2.2.2-trifluoroethyl)-lH-benzoleiri .41diazepin-3-yl1acetamide
Figure imgf000153_0002
m.p. 172-173 °C, [α]D = +5.9° (c=0.56, CHC13). Anal. Calcd. for C26H19F6N3O2O.6O H20:
C, 58.89; H, 3.84; N, 7.92. Found: C, 58.92; H, 3.71; N, 7.98%.
EXAMPLE 118
(+)-2-(2-Trifluoromethylphenyl)-N-[3R-2,3-dihydro 2-0X0-5- phenyl-l-(2.2.2-trifluoroethyl)-lH-benzoleiπ .41diazepin-3-yllacetamide
Figure imgf000154_0001
m.p. 170-171 °C, [α]D = +9.0° (c=0.48, CHCI3). Anal. Calcd. for C_6Hl9F6N3θ2-0.25 H20:
C, 59.6; H, 3.75; N, 8.02. Found: C, 59.64; H, 3.68; N, 7.97%.
EXAMPLE119
(-)-2-(2,4-Dichlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5-phenyl- l-(2.2.2-trifluoroethviy lH-benzofelf 1.41diazepin-3-yllacetamide
Figure imgf000154_0002
m.p. 143-145 °C, [α]D = -22.6° (c=0.73; MeOH). Anal. Calcd. for C25HI 8N3O2CI2F3:
C, 57.71 ; H, 3.49; N, 8.08. Found: C, 57.75; H, 3.52; N, 8.09%.
EXAMPLE 120
(-)-2-(3-Chlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5-phenyl- 1 - (2.2.2-trifluoroethvD-l H-benzolel \ 1.41diazepin-3-yllacetamide
Figure imgf000155_0001
m.p. 188-189 °C, [α]D = -5.4° (c= 1.03, MeOH). Anal. Calcd. for C25H19C.F3N3O2-O.IO ethyl ether:
C, 61.84; H, 4.09; N, 8.52. Found: C, 61.84; H, 4.05; N, 8.5%.
EXAMPLE 121
(-)-2-(4-Chlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5-phenyl- l -(2.2.2-trifluoroethyl)-lH-benzore111.41diazepin-3-yllacetamide
Figure imgf000156_0001
m.p. 246-247 °C, [α]D =-10.1° (c=0.45,MeOH).
Anal. Calcd. for C25H19CIF3N3O2O.2O H20 0.15 ethyl ether:
C, 61.42; H, 4.21; N, 8.39. Found: C, 61.46; H, 4.15; N, 8.39%.
Example 122
(-)-2-[2,4-Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5- phenyl-l-(2.2.2-trifluoroethylVlH-benzoleiπ.41diazepin-3-yllacetamide
Figure imgf000156_0002
Step A. 2,4-Bis(trifluoromethyl)benzonitrile
To a stirring biphasic mixture of lOOmL ethanol and 250 mL of phosphate buffer (lg of NaH2Pθ4»H2θ per 5 mL H2O adjusted to pH=7.0 with 50% NaOH) and NaCN (81.3mmol,4.0g) heated to 60°C was added 2,4-bis(trifluoromethyl) benzyl bromide (32.5mmol,10g) in 50mL EtOH dropwise over 30min. The reaction was heated at 60°C for 24h. The reaction was then evaporated under reduced pressure. The remaining aqueous was extracted with 2xl50mL EtOAc. The organic layers were combined, dried with brine and Na2Sθ4. The organic phase was evaporated under reduced pressure and the residue chromatographed over silica eluting with 10% EtOAc:Hexanes. The pure fractions were collected and evaporated to give 7.0g of a pale yellow oil, 85.1 % NMR H (CDCI3) d 8.0-7.85 (m,3H), 4.03 (s,2H)
Step B. 2,4-Bis(trifluoromethyl)phenyl acetic acid
2,4-Bis(trifluoromethyl)benzonitrile (41.5mmol, 10.51 g) was taken up in lOOmL acetic acid, 50mL cone. H2SO4, and 20mL water. This was heated to 120°C for 3h. The reaction was then diluted with 1L ice water, and extracted with 2x300mL ethyl acetate. The combined organics were washed with 2x200mL water, dried with brine and Na2S04, and evaporated under reduced pressure. The residue was taken up in a minimum of diethyl ether and crystallized by adding sufficient hexane to precipatate the product. The solid was collected to give 7.74g of 2,4-bis(trifiuoromethyl) phenyl acetic acid as white crystals, 68.5 %.NMR JH (CDCI3) d 7.93 (s,lH), 7.80 (d, J=7.9Hz, lH), 7.55 (d, J=7.9Hz,lH), 3.94 (s,2H).
Step C. Preparation of (-)-2-[2,4-Bis(trifluoromethyl)phenyl]-N-
[3R-2,3-dihydro-2-oxo-5-phenyl- 1 -(2,2,2-trifluoroethy 1)- 1 H-benzo[e][ 1 ,4]diazepin-3-yl]acetamide
To a stirring solution the 3R-3-Amino-2-oxo-5-phenyl-l - (2,2,2-trifluoroethyl)-2,3-dihydro-lH-benzo[e][l ,4]diazepine (28.4 mmol, 9.47g) in DMF (lOOmL) was added 2,4-Bis(trifluoromethyl) phenyl acetic acid (28.4mmol,7.74g), 1 -(3-Dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (42.6mmol,8.16g) and 1-Hydroxy- benztriazole hydrate (14.2mmol,1.92g). This was stined for lh at room temperature. The reaction was then diluted with 750mL of 10% KHSO4 and extracted with ethyl acetate (2x300mL). The organic layers were combined and washed with saturated sodium hydrogen carbonate (lx600mL). The organics were then dried with brine, and sodium sulfate and evaporated under reduced pressure. The residue was chromatographed over silica eluting with 20% EtOAc:Hexane. Pure fractions were collected and evaporated. The residue was taken up in 100 mL of warm 75% isopropanol: water. This was allowed to cool slowly and stirred overnight (16 hr) at room temperature. The suspension was cooled briefly to @5°C and filtered. The white solid was dried overnight at 60°C to give 10.5 g of material that melted at 132-134°C. X-Ray diffraction confirms crystal linitv . NMR lH (CDCl3) d 7.95-7.25 (m,13H), 5.60 (d,J=8.1 Hz, IH), 5.30- 5.10 (m,lH), 4.25-4.06 (m.lH), 3.96 (s,2H) Anal. Calcd. for C27HI 8F9N3O2:
C, 55.20; H, 3.09; N, 7.15. Found: C, 55.03; H, 3.14; N, 7.10%.
EXAMPLE 123
(+)-2-(3,5-Dichlorophenyl)-N-[2,3-dihydro-2-oxo-5-phenyl- 1 -(2.2.2-trifluoroethviy 1 H-benzolel [ 1 ,41diazepin-3-yll acetamide
Figure imgf000158_0001
m.p. 219-220 °C. racemic
Anal. Calcd. for C25HI 8N3O2CI2F3:
C, 57.71; H, 3.49; N, 8.08. Found: C, 57.94; H, 3.48; N, 8.02%. EX AMPLE 124
2-(3,5-dichloro-4-me oxyphenyl)-N-[3R-2,3-dihydro-2-oxo-5- phenyl- 1 -(2.2.2-trifluoroethylV 1 H-benzorel \ 1.41diazepin-3-yllacetamide
Figure imgf000159_0001
m.p. 100-104 °C, [α]D = -8.9° (c=0.55,MeOH). Anal. Calcd. for C26H20CI2F3N3O3:
C, 56.74; H, 3.66; N, 7.63. Found: C, 55.67; H, 3.47; N, 7.41%.
The following examples were prepared by procedures substantially as described in example 1 except substituting the appropriate fluoro substituted aminobenzophenone in step A.
EXAMPLE 125
(+)-2-(3,5-Dichlorophenyl)-N-[2,3-dmydro-5-(4-fluorophenyl)-2- oxo- 1 -(2.2.2-trifluoroethyl)- 1 H-benzolel \ 1.41diazepin-3-yllacetamide
Figure imgf000160_0001
m.p. foam °C, [α]D = +3.4° (c=0.55; MeOH). Anal. Calcd. for C25H17N3O2CI2F4:
C, 55.78; H, 3.18; N, 7.81. Found: C, 55.73; H, 3.25; N, 7.72%.
EXAMPLE 126
(.)-2-(2,4-Dichlorophenyl)-N-[2,3-dihydro-5-(4-flu oxo-1 -(2.2.2-trifluoroethyl)-l H-benzofelf 1 ,41diazepin-3-yllacetamide
Figure imgf000160_0002
m.p. foam °C, [α]D = -11° (c=0.68; MeOH). Anal. Calcd. for C25H17N3O2F4:
C, 55.78; H, 3.18; N, 7.81. Found: C, 55.82; H, 3.41; N, 7.42%. EX AMPLE 127
(+)-2-(3,5-Bis(trifluoromethyl)phenyl)-N-[2,3-dihydro-5-(4- fluoroρhenyl)-2-oxo- 1 -(2,2,2-trifluoroethyl)-lH-benzo[e]
11.41diazepin-3-yll-acetamide
Figure imgf000161_0001
m.p. foam °C, [a]D = +2.8° (c=0.67; MeOH). Anal. Calcd. for C27H17N3O2F1O:
C, 53.56; H, 2.83; N, 6.94. Found: C, 53.56; H, 2.93; N, 6.91%.
EXAMPLE 128
(-)-2-[2,4-Bis(trifluoromethyl)phenyl]-N-[2,3-dihydro-5-(4- fluorophenyl)-2-oxo 1 -(2,2,2-trifluoroethyl)- 1 H-benzo[e] [ 1 ,4] diazepin-3-yllacetamide
Figure imgf000162_0001
[α]D = -14° (c=0.63; MeOH). Anal. Calcd. for C27H17N3O2F10:
C, 53.56; H, 2.83; N, 6.94. Found: C, 53.3; H, 2.89; N, 7.05%.
EXAMPLE 129
3-Cyclohexyl-N-[2,3-dihydro-5-(2-fluorophenyl)-2-oxo- 1 - (2.2.2-trifluoroethyl-lH-benzorelll .41diazepin-3-yllpropionamide
m.p. 202-204 °C. iH NMR d (CDC13) 7.72 (m,8H), 5.65 (d,J=8.3Hz,lH), 5.35-5.08 (m,lH), 4.32-4.15 (m,lH), 2.37 (t,J=7.8Hz,2H), 1.80-1.55 (m,7H), 1.45-Anal. Calcd. for C26H27F4N3O2: C, 63.8; H, 5.56; N, 8.58. Found: C, 63.82; H, 5.54; N, 8.56%.
EXAMPLE 130
3,4-Dichloro-N-[2,3-dihydro-5-(2-fluorophenyl)-2-oxo-l - (2.2.2-trifluoroethyl')- 1 H-benzorel 1.41diazepin-3-yllbenzamide
Figure imgf000163_0001
m.p. 168-170 °C. iH NMR d (CDC13) 8.03 (d,J=2.0,lH), 7.86 (d,J=7.8Hz,lH), 7.78-7.05 (m,9H), 5.80 (d,J=7.8Hz,lH), 5.27-5.15 (m,l H), 4.35-4.20 (m,lH) Anal. Calcd. for C24H15CI2F4N3O2:
C, 54.98; H, 2.88; N, 8.01. Found: C, 54.96; H, 2.89; N, 8.12%.
EXAMPLE 131
Antianhythmic efficacy of the combined administration of low dose Ij s blocker (-)-2-[2,4-bis(trifluoromethyl)phenyl]-n-[3r-2,3- dihydro-2-oxo-5-phenyl-l -(2,2,2-trifluoroethyl)-lh-benzo[e][l ,4] diazepin-3-yl]acetamide and the beta-adrenergic receptor antagonist timolol in anesthetized dogs with previous anterior myocardial infarction was studied. This study was conducted to t st the hypothesis the concomitant adminis-tration of low dose Ij^s blocker (-)-2-[2,4-
Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5-phenyl- 1 - (2,2 ,2-trifluoroethy 1)- 1 H-benzo [e] [ 1 ,4] diazepin- 3 -y 1] acetamide and low dose beta-adrenergic receptor antagonist timolol, which when administered individually at low dose afford no significant anti- anhythmic protection, would in combination provide significant protection against the development of malignant ventricular anhythmias in a canine model of previous myocardial infarction. In the absence of protective intervention, this postinfarction canine preparation displays a very high incidence ( 80%) of malignant ventricular tachycardia degenerating into ventricular fibrillation and death in response to the development of ischemia at a site remote from previous myocardial infarction (Patterson et al., Am. J. Cardiol. 50: 1414-1423, 1982; Wilber et al., Am. Heart J. 109: 8-18, 1985; Lynch et al., J Pharmacol Exp Therap 277: 671-678, 1996).
METHODS
Surgical Preparation.
Male or female purpose-bred mongrel dogs (7-12 kg) were preanesthetized with sodium thiamylal (5.0 mg/kg i.v.) and general anesthesia was induced with isoflurane. A left thoracotomy was performed in the 4th intercostal space, the pericardium incised, and the heart suspended in a pericardial cradle. Anterior myocardial infarction was produced by a two hour occlusion of the left anterior descending coronary artery followed by reperfusion. Surgical incisions were closed, and the animals were allowed to recover.
Electrophysiologic Testing, Programmed Ventricular Stimulation (PVS) and Acute Posterolateral Myocardial Ischemia .
Purpose-bred male or female mongrel dogs at 5-20 days after anterior myocardial infarction were anesthetized with alpha chloralose (80.0-100.0 mg/kg i.v.). The animals were ventilated by means of a cuffed endotracheal tube and a volume-cycled respirator (Model 613, Harvard Apparatus, S. Natick, MA). Systemic arterial pressure was monitored via the cannulated left common carotid artery (Model P23XL pressure transducer, Gould Inc, Cleveland, OH), and the right femoral vein was isolated and cannulated for drug administration. The heart was re-exposed via a left thoracotomy, and was suspended in a pericardial cradle. A platinum epicardial bipolar electrode (3 mm electrode post separation) was sutured to the surface of the left atrial appendage for atrial pacing, and a stainless steel bipolar plunge electrode (5 mm length, 3 mm electrode post separation) was inserted into the interventricular septum near the right ventricular outflow tract (RVOT) adjacent to the site of left anterior descending coronary artery occlusion for the introduction of ventricular extrastimuh during programmed ventricular stimulation (PVS). One acrylic button per zone containing fixed stainless steel bipolar plunge electrodes (see below for specifications) was sutured into the infarcted anterior region of the left ventricle distal to the site of coronary artery occlusion and within the area of myocardial scarring as ascertained visually and by palpation (IZ, infarct zone) and into the non-infarcted posterolateral region of the left ventricle (NZ, non-infarct zone) for the measurement of ventricular electrophysiologic parameters (excitation thresholds and refractory periods). Lead II electrocardiogram was monitored continuously.
After stabilization of the preparation and the measurement of baseline electrocardiographic and cardiac electrophysiologic parameters, PVS consisting of the introduction of 1-3 ventricular extrastimuh during sinus rhythm and atrial pacing was performed at the RVOT site. If ventricular extrastimuh could not be inserted at the RVOT site, programmed pacing was attempted at the IZ site. Ventricular extrastimuh were introduced at 2x diastolic threshold voltage or, if extrastimuh were not introduced adequately, at 4x diastolic threshold voltage. Responses to baseline programmed ventricular stimulation were categorized as: 1) non-inducible (NI): less than 5 nonstimulated ventricular complexes; 2) nonsustained ventricular tachycardia (NSVT): 5 or more nonstimulated ventricular complexes terminating spontaneously with a duration less than 15 seconds; 3) unimorphic sustained ventricular tachycardia (UVT): unimorphic ventricular tachycardia with a duration exceeding 15 seconds; 4) polymorphic sustained ventricular tachycardia (PVT): polymorphic ventricular tachycardia with a duration exceeding 15 seconds, and 5) ventricular tachycardia degenerating into ventricular fibrillation (VT/VF): an unstable ventricular tachycardia degenerating spontaneously into ventricular fibrillation. In this study, programmed pacing was continued until the induction of either sustained VT or VT/VF, or until the end of the pacing protocol with either NSVT or no response (NI) occurring. In many preparations ultimately responding to programmed stimulation with either SVT or VT/VF, reproducible NSVTs were initiated early in the pacing protocol. Only postinfarction preparations responding to baseline PVS with inducible sustained ventricular tachyanhythmias (UVT, PVT or VT/VF) were entered into the present study. Baseline response to PVS with inducible ventricular tachyarrhythmias has been found to correlate strongly with the development malignant ventricular tachycardia, fibrillation and anhythmic death in response to a subsequent episode of myocardial ischemia at a site remote from previous infarction in this animal model (Wilber et al., Am. Heart J. 109: 8-18, 1985).
Following the completion of baseline and post treatment electrophysiologic and PVS testing, the tip of a silver wire electrode was inserted through the wall and into the lumen of the proximal left circumflex (LCX) coronary artery. An anodal cunent of 200 μA was applied to the intimal surface of the coronary artery via this electrode, producing intimal injury, thrombus formation and ultimately acute myocardial ischemia in the left circumflex coronary artery distribution. The onset of acute posterolateral myocardial ischemia was noted electro- cardiographically. Upon the development of lethal ischemic anhythmias or at 3 hours after the onset of acute posterolateral myocardial ischemia, the hearts were excised and wet thrombus mass in the left circumflex coronary artery determined. Anterior myocardial infarct size was determined by cutting the heart into 1 cm thick transverse sections which then were incubated in 0.4% triphenyltetrazolium chloride solution. Reaction with triphenyltetrazolium forms a red precipitate in viable tissue, whereas infarcted tissue remains pale. Infarct size was quantitated gravimetrically and was expressed as a percentage of total left ventricle.
The responses (i.e. occunence of vs. protection from the development of lethal ventricular anhythmia) of five experimental groups to the development of acute posterolateral ischemia at a site remote from previous myocardial infarction were compared in this study: 1 ) the laboratory cohort of aqueous vehicle (saline or PEG/ saline)-treated control animals (n=40); 2) a dedicated soy bean oil-based microemulsion vehicle (20.0% soy bean oil, 2.0% glycerin, 1.2% lecithin and 76.8% water)-treated control group (n=10); 3) low dose 0.0003 mg/kg i.v. IKs blocker (-)-2-[2,4-Bis(trifluoromethyl)phenyl]-N-
[3R-2,3-dihydro-2-oxo-5-phenyl-l-(2,2,2-trifluoroethyl)-lH- benzo[e][l,4]diazepin-3-yl]acetamide in soy bean oil-based microemulsion (n=10); 4) low dose 0.001 mg/kg i.v. beta-adrenergic receptor blocking agent timolol in 5% dextrose in saline vehicle (n=6); and 5) the combination of low dose 0.0003 mg/kg i.v. Ij s blocker
(-)-2-[2,4-Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihvdro-2-oxo-5- phenyl-l-(2,2,2-trifluoroethyl)-lH-benzo[e][l ,4]diazepin-3-yl]acetamide with low dose 0.001 mg/kg i.v. beta-adrenergic receptor blocking agent timolol (n=10). In the individual and combination Ij^s blocker (-)-2- t2,4-Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5-phenyl-l- (2,2,2-trifluoroethyl)-lH-benzo[e][ 1 ,4]diazepin-3-yl]acetamide and beta- adrenergic receptor blocking agent timolol treatment groups, treatments were administered i.v. 15 min prior to the conduct of post treatment PVS and production of ischemia at a site remote from previous infarction.
Statistics.
Data are expressed as mean ± S.E.M. Comparisons among treatment groups were made using a single factor ANOVA or a Fisher's exact test, when appropriate. P 0.05 was the criterion for statistical significance. RESULTS
All animals in all treatment groups responded to baseline PVS with inducible ventricular tachyarrhythmias as an entry criterion. The incidences of ischemic lethal ventricular anhythmias (ventricular fibrillation; VF) occurring in the laboratory control cohort, dedicated microemulsion vehicle control group, low dose Ij s blocker (-)-2-[2,4-
Bis(trifluoromethyl)phenyll-N-[3R-2,3-dihydro-2-oxo-5-phenyl-l- (2,2,2-trifluoroethyl)-lH-benzo[e][l ,4]diazepin-3-yl]acetamide, low dose beta-adrenergic receptor blocking agent timolol, and combined low dose IKs blocker (-)-2-[2,4-Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-
2-oxo-5-phenyl- 1 -(2,2,2-trifluoroethyl)- lH-benzo[e][ 1 ,4]diazepin-3- yljacetamide with beta-adrenergic receptor blocking agent timolol rou s are summarized below.
Figure imgf000168_0001
NS = nonsignificant (P>0.05). Neither low dose I| s blocker (-)-2-[2,4- Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5-phenyl- 1 - (2,2,2-trifluoroethyl)-lH-benzo[e][l ,4]diazepin-3-yl]acetamide nor low dose beta-adrenergic receptor antagonist timolol administered individually afforded significant protection against the development of malignant ischemic ventricular anhythmias and death compared to either the laboratory vehicle control cohort or the dedicated microemulsion vehicle control group. In contrast, the combined administration of both low dose I^s blocker (-)-2-[2,4-
Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5-phenyl- 1 - (2,2,2-trifluoroethyl)-lH-benzo[e][ 1 ,4]diazepin-3-yl]acetamide and low dose beta-adrenergic receptor antagonist timolol provided significant protection against development of malignant ischemic ventricular tachyanhythmia and anhythmic death, which signifies a synergistic salutary antianhythmic interaction between block of I^s and beta- adrenergic receptor blockade.

Claims

WHAT IS CLAIMED IS:
1. A method of treating anhythmia which comprises the co-administration to a patient in need of such treatment of an effective amount of a beta-adrenergic receptor blocking agent and a selective Ij s antagonist.
2. The method of Claim 1 wherein the selective Ijζs antagonist is a compound of structural formula:
Figure imgf000170_0001
individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt thereof, wherein
A is
1 ) thieno,
2) pyrido, or
3) benzo either unsubstituted or substituted with -NH2, -NHS02(Cl-3 alkyl), Cl-3 alkyl or Cl-3 alkoxy;
X is
Figure imgf000170_0002
Y is
1) = o,
2) = N-CN or
3) = H2;
Z is
1) Cl-6 alkylene, either straight or branch chain and either unsubstituted or substituted with phenyl or spiropiperidine,
2) C2-4 alkenylene, either straight or branch chain,
3) -(CH2)m- -(CH2)n- wherein m and n are independently 0, 1, 2, 3 or 4 and W is -0-, -S- or -NH,
4) 4-(5-methylisoxazole-3-yl),
5) C3-6 cycloalkylene, or
6) single bond;
p is 0 or 1 ;
Figure imgf000171_0001
4) mono- or bicyclic heterocyclyl of 5 to 1 members one or two of which are sulfur, nitrogen or oxygen, the remaining being carbon, such as 2-thienyl, 2-furanyl, 2-indolyl, 2- quinoxolinyl, or 2-(2,3-dihydro benzofuranyl)
5) methyl, or
6) indan-5-yl;
R2 IS
1) phenyl, either unsubstituted or substituted with Cl-3 alkoxy or 4,4-dimethyloxazolin-2-yl,
2) Cl -4 alkyl, either straight or branched chain and either unsubstituted or substituted with Cl-3 alkoxy or
Cl-3 alkoxy-Cl-3 alkoxy,
3) C5-7 cycloalkyl,
4) 2- or 3-furyl,
5) l -methylpiperidin-2-yl, or
6) if R2 is phenyl, the 2-position of the phenyl can be joined to the 4-position nitrogen of the diazepine ring through a carbonyl group and the double bond between the 4-nitrogen and the 5-carbon becomes a single bond;
1 ) hydrogen or
2) Cl-6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3, or
3) -CF3;
R4 IS
1 ) hydrogen,
2) Cl-6 alkyl, the chain of carbon atoms of which can be interrupted by one or two non-adjacent oxygen atoms and which is either unsubstituted or substituted with Cl-3 alkoxycarbonyl, -OH or
Figure imgf000172_0001
3) tetrazol-5-yl; and R5 is hydrogen or oxygen or is joined to R2 to form the partial structure:
Figure imgf000173_0001
and the bond represented by .____ is:
1 ) a double bond when p is zero or when p is 1 and R^ is oxygen, or
2) a single bond when R^ is hydrogen or R^ is joined to R^ to form the partial structure:
Figure imgf000173_0002
3. The selective Ijζs compound of Claim 2 wherein
A is benzo either unsubstituted or substituted with -NH2, -NHS02(Cl -3 alkyl), Cl-3 alkyl or Cl -3 alkoxy;
X and Y are O, Z is
1) Cl-6 alkylene, either straight or branch chain and either unsubstituted or substituted with phenyl or spiropiperidine,
2) C2-4 alkenylene, either straight or branch chain,
3) -(CH2)m-W-(CH2)n- wherein m and n are independently 0, 1 , 2, 3 or 4 and W is -0-, -S- or -NH,
4) 4-(5-methylisoxazole-3-yl),
5) C3-6 cycloalkylene, or 6) single bond;
Rl is
Figure imgf000174_0002
Figure imgf000174_0001
4) mono- or bicyclic heterocyclyl of 5 to 10 members one or two of which are sulfur, nitrogen or oxygen, the remaining being carbon, such as 2-thienyl, 2-furanyl, 2-indolyl, 2- quinoxolinyl, or 2-(2,3-dihydro benzofuranyl)
5) methyl, or
6) indan-5-yl;
1 ) phenyl, either unsubstituted or substituted with Cl-3 alkoxy or 4,4-dimethyloxazolin-2-yl,
2) Cl-4 alkyl, either straight or branched chain and either unsubstituted or substituted with Cl-3 alkoxy or
Cl-3 alkoxy-Ci-3 alkoxy,
3) C5-7 cycloalkyl,
4) 2- or 3 -fury 1,
5) l-methylpiperidin-2-yl, or 6) if R2 is phenyl, the 2-position of the phenyl can be joined to the 4-position nitrogen of the diazepine ring through a carbonyl group and the double bond between the 4-nitrogen and the 5-carbon becomes a single bond;
R3 IS
1 ) hydrogen or
2) Cl -6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3, or
3) -CF3;
R i is
1 ) hydrogen,
2) Cl-6 alkyl, the chain of carbon atoms of which can be interrupted by one or two non-adjacent oxygen atoms and which is either unsubstituted or substituted with Cl -3 alkoxy carbonyl, -OH or
Figure imgf000175_0001
3) tetrazol-5-yl; and
R5 is hydrogen or oxygen or is joined to R^ to form the partial structure:
Figure imgf000175_0002
and the bond represented by ^n is:
1 ) a double bond when p is zero or when p is 1 and R5 is oxygen, or
2) a single bond when R^ is hydrogen or R5 is joined to R2 to form the partial structure:
Figure imgf000176_0001
including individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt.
4. The selective I^s compound of Claim 3 wherein
A is benzo either unsubstituted or substituted with -NH2, -NHSθ2(Cl-3 alkyl), Cl-3 alkyl or Cl -3 alkoxy;
X and Y are O,
Z is
1 ) Cl-6 alkylane, either straight or branch chain and either unsubstituted or substituted with phenyl or spiropiperidine,
2) C2-4 alkenylene, either straight or branch chain,
3) -(CH2)m-W-(CH2)n- wherein m and n are independently 0, 1, 2, 3 or 4 and W is -O-, -S- or -NH,
4) 4-(5-methylisoxazole-3-yl),
5) C3-6 cycloalkylene, or
6) single bond;
R 1 is phenyl, either unsubstituted or substituted with one or two substituents selected from a) -Nθ2, b) -CI, Br, F, or I, c) -CF3, d) -Cl-3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy, R2 is phenyl, either unsubstituted or substituted with Cl -3 alkoxy or
4,4-dimethyloxazolin-2-yl, R3 is -CF3 or Cl -6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3;
R4 and R5 are hydrogen;
including individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt.
5. The selective Iχs compound of Claim 4 wherein A is unsubstituted benzo;
Rl is phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02, b) -CI, Br, F, or I, c) -CF3, d) -Cl -3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy,
R2 is phenyl, either unsubstituted or substituted with Cl-3 alkoxy or 4,4-dimethyloxazolin-2-yl,
R is -CF3 or Cl-6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3;
R4 and R^ are hydrogen;
including individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt.
6. The selective Iχ. s compound of Claim 5 which is (-)- 2-[2,4-Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5-phenyl- 1 -(2,2,2-trifluoroethyl)- 1 H-benzo[e][ 1 ,4]diazepin-3-yl]acetamide
Figure imgf000178_0001
7. The selective Ijςs compound of Claim 5 which is 3,4- Dichloro-N-[2,3-dihydro-5-(2-fluorophenyl)-2-oxo-l-(2,2,2-trifluoro- ethyl)-lH-benzo[e][ 1 ,4]diazepin-3-yl]benzamide
Figure imgf000178_0002
8. The selective Ij s compound of Claim 5 which is (-)-2-(2,4-Dichlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5-phenyl-l- (2,2,2-trifluoroethyl)- 1 H-benzo[e] [ 1 ,4]diazepin-3-yl]acetamide
Figure imgf000179_0001
8. The beta-adrenergic receptor blocking agent of Claim 1 selected from the group consisting of timolol, sotalol, esmolol, cateolol, propranolol, betaxolol, penbutolol, metoprolol, acebutolol, atenolol, metoprolol, pindolol, or bisoprolol.
9. A pharmaceutical formulation comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a beta-adrenergic receptor blocking agent and a selective Ij^s antagonist.
10. The beta-adrenergic receptor blocking agent of Claim 9 selected from the group consisting of timolol, sotalol, esmolol, cateolol, propranolol, betaxolol, penbutolol, metoprolol, acebutolol, atenolol, metoprolol, pindolol, or bisoprolol.
11. The selective Ij s antagonist of Claim 9 which is a compound of structural formula:
Figure imgf000179_0002
individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt thereof, wherein
A is
1 ) thieno,
2) pyrido, or
3) benzo either unsubstituted or substituted with -NH2, -NHS02(Cl-3 alkyl), Cl-3 alkyl or Cl-3 alkoxy;
Figure imgf000180_0001
Z is
1 ) Cl-6 alkylene, either straight or branch chain and either unsubstituted or substituted with phenyl or spiropiperidine,
2) C2-4 alkenylene, either straight or branch chain,
3) -(CH2)m- -(CH2)n- wherein m and n are independently 0, 1 , 2, 3 or 4 and W is -0-, -S- or -NH,
4) 4-(5-methylisoxazole-3-yl),
5) C3-6 cycloalkylene, or
6) single bond;
p is 0 or 1 ; Rl i IS
Figure imgf000181_0002
Figure imgf000181_0001
4) mono- or bicyclic heterocyclyl of 5 to 10 members one or two of which are sulfur, nitrogen or oxygen, the remaining being carbon, such as 2-thienyl, 2-furanyl, 2-indolyl, 2- quinoxolinyl, or 2-(2,3-dihydro benzofuranyl)
5) methyl, or
6) indan-5-yl;
R2 IS
1 ) phenyl, either unsubstituted or substituted with Cl -3 alkoxy or 4,4-dimethyloxazolin-2-yl,
2) Cl-4 alkyl, either straight or branched chain and either unsubstituted or substituted with Cl-3 alkoxy or
Cl -3 alkoxy-Ci -3 alkoxy,
3) C5-7 cycloalkyl,
4) 2- or 3-furyl,
5) 1 -methy lpiperidin-2-yl, or
6) if R2 is phenyl, the 2-position of the phenyl can be joined to the 4-position nitrogen of the diazepine ring through a carbonyl group and the double bond between the 4-nitrogen and the 5-carbon becomes a single bond: R3 i IfS
1) hydrogen or
2) Cl-6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3, or
3) -CF3;
R4 IS
1 ) hydrogen,
2) Cl-6 alkyl, the chain of carbon atoms of which can be interrupted by one or two non-adjacent oxygen atoms and which is either unsubstituted or substituted with Cl-3 alkoxycarbonyl, -OH or
Figure imgf000182_0001
3) tetrazol-5-yl; and
R5 is hydrogen or oxygen or is joined to R2 to form the partial structure:
Figure imgf000182_0002
the bond represented by ____; is:
1 ) a double bond when p is zero or when p is 1 and R^ is oxygen, or
2) a single bond when R^ is hydrogen or R^ is joined to R2 to form the partial structure:
Figure imgf000183_0001
12. The selective Iκs compound of Claim 1 1 wherein
A is benzo either unsubstituted or substituted with -NH2, -NHS02 (Cl -3 alkyl), Cl-3 alkyl or Cl -3 alkoxy;
X and Y are O,
Z is
1 ) Cl -6 alkylene, either straight or branch chain and either unsubstituted or substituted with phenyl or spiropiperidine,
2) C2-4 alkenylene, either straight or branch chain,
3) -(CH2)m- -(CH2)n- wherein m and n are independently 0, 1 , 2, 3 or 4 and W is -0-, -S- or -NH,
4) 4-(5-methylisoxazole-3-yl),
5) C3-6 cycloalkylene, or
6) single bond;
Rl is
1 ) phenyl, either unsubstituted or substituted with one or two substituents selected from a) -Nθ2, b) -CI, Br, F, or I, c) -CF3, d) -Cl -3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy,
2) C5-7 cycloalkyl, 3)
Figure imgf000184_0001
4) mono- or bicyclic heterocyclyl of 5 to 10 members one or two of which are sulfur, nitrogen or oxygen, the remaining being carbon, such as 2-thienyl, 2-furanyl, 2-indolyl, 2- quinoxolinyl, or 2-(2,3-dihydro benzofuranyl)
5) methyl, or
6) indan-5-yI;
1 ) phenyl, either unsubstituted or substituted with Cl-3 alkoxy or 4,4-dimethyloxazolin-2-yl,
2) Cl -4 alkyl, either straight or branched chain and either unsubstituted or substituted with Cl-3 alkoxy or
Cl -3 alkoxy-Cl-3 alkoxy,
3) C5-7 cycloalkyl,
4) 2- or 3-furyl,
5) 1 -methy lpiperidin-2-yl, or
6) if R2 is phenyl, the 2-position of the phenyl can be joined to the 4-position nitrogen of the diazepine ring through a carbonyl group and the double bond between the 4 -nitrogen and the 5-carbon becomes a single bond;
R3 i is
1) hydrogen or
2) Cl-6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3, or
3) -CF3;
R4 is
1 ) hydrogen,
2) Cl-6 alkyl, the chain of carbon atoms of which can be interrupted by one or two non-adjacent o ygen atoms and which is either unsubstituted or substituted with Cl-3 alkoxycarbonyl, -OH or
Figure imgf000185_0001
3) tetrazol-5-yl; and
R5 is hydrogen or oxygen or is joined to R2 to form the partial structure:
Figure imgf000185_0002
and the bond represented by ____ is:
1 ) a double bond when p is zero or when p is 1 and R^ is oxygen, or
2) a single bond when R^ is hydrogen or R5 is joined to R to form the partial structure:
Figure imgf000185_0003
including individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt.
13. The selective Ij s compound of Claim 12 wherein A is benzo either unsubstituted or substituted with -NH2, -NHSθ2(Cι_3 alkyl), Cl-3 alkyl or Cl -3 alkoxy;
X and Y are O,
Z is 1 ) Cl-6 alkylane, either straight or branch chain and either unsubstituted or substituted with phenyl or spiropiperidine,
2) C2-4 alkenylene, either straight or branch chain,
3) -(CH2)m-W-(CH2)n- wherein m and n are independently 0, 1, 2, 3 or 4 and W is -0-, -S- or -NH,
4) 4-(5-methylisoxazole-3-yl),
5) C3-6 cycloalkylene, or
6) single bond;
Ri is phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02, b) -CI, Br, F, or I, c) -CF3, d) -Cl -3 alkyl, e) -Cl -3 alkoxy, f) -CN, g) -methylenedioxy,
R is phenyl, either unsubstituted or substituted with Cl-3 alkoxy or 4,4-dimethyloxazolin-2-yl,
R3 is -CF3 or Cl -6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3;
R4 and R^ are hydrogen;
including individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt.
14. The selective Ij s compound of Claim 13 wherein A is unsubstituted benzo; R! is phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02, b) -CI, Br, F, or I, c) -CF3, d) -Cl-3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy,
R is phenyl, either unsubstituted or substituted with Cl-3 alkoxy or 4,4-dimethyloxazolin-2-yl,
R3 is -CF3 or Cl-6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3;
R4 and R5 are hydrogen;
including individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt.
15. The selective Ijζs compound of Claim 14 which is (-)-2-[2,4-Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5- phenyl- 1 -(2,2,2-trifluoroethyl)- lH-benzo[e][ 1 ,4]diazepin-3-yl]acetamide
Figure imgf000187_0001
16. The selective Ij s compound of Claim 14 which is 3,4-Dichloro-N-[2,3-dihydro-5-(2-fluorophenyl)-2-oxo-l -(2,2,2- trifluoroethyl)- 1 H-benzo[e] [ 1 ,4]diazepin-3-yl]benzamide
Figure imgf000188_0001
17. The selective Ijζs compound of Claim 14 which is (-)-2-(2,4-Dichlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5-phenyl-l- (2,2,2-trifluoroethyl)- 1 H-benzo[e] [ 1 ,4]diazepin-3-yl]acetamide
Figure imgf000188_0002
18. A method of preventing anhythmia which comprises the co-administration to a patient in need of such treatment of an effective amount of a beta-adrenergic receptor blocking agent and a selective Ij£s antagonist.
19. The method of Claim 18 wherein the selective Ijζs antagonist is a compound of structural formula:
Figure imgf000189_0001
individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt thereof, wherein
A is
1 ) thieno,
2) pyrido, or
3) benzo either unsubstituted or substituted with -NH2, -NHSθ2(Cl -3 alkyl), Cl-3 alkyl or Cl-3 alkoxy;
Figure imgf000189_0002
Z is
1 ) Cl -6 alkylene, either straight or branch chain and either unsubstituted or substituted with phenyl or spiropiperidine,
2) C2-4 alkenylene, either straight or branch chain, 3) -(CH2)m-W-(CH2)n- wherein m and n are independently 0, 1 , 2, 3 or 4 and W is -0-, -S- or -NH,
4) 4-(5-methylisoxazole-3-yl),
5) C3- cycloalkylene, or
6) single bond;
p is 0 or 1 ;
Figure imgf000190_0001
4) mono- or bicyclic heterocyclyl of 5 to 10 members one or two of which are sulfur, nitrogen or oxygen, the remaining being carbon, such as 2-thienyl, 2-furanyl, 2-indolyl, 2- quinoxolinyl, or 2-(2,3-dihydro benzofuranyl)
5) methyl, or
6) indan-5-yl;
R2 IS
1 ) phenyl, either unsubstituted or substituted with Ci _3 alkoxy or 4,4-dimethyloxazolin-2-yl,
2) Cl-4 alkyl, either straight or branched chain and either unsubstituted or substituted with Cl-3 alkoxy or Cl -3 alkoxy-Ci -3 alkoxy,
3) C5-7 cycloalkyl,
4) 2- or 3-furyl,
5) 1 -methylpiperidin-2-yl, or
6) if R2 is phenyl, the 2-position of the phenyl can be joined to the 4-position nitrogen of the diazepine ring through a carbonyl group and the double bond between the 4-nitrogen and the 5 -carbon becomes a single bond;
R3 i is
1) hydrogen or
2) Cl-6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3, or
3) -CF3;
R4 IS
1 ) hydrogen,
2) Cl-6 alkyl, the chain of carbon atoms of which can be interrupted by one or two non-adjacent oxygen atoms and which is either unsubstituted or substituted with Cl-3 alkoxy carbonyl, -OH or
Figure imgf000191_0001
3) tetrazol-5-yl; and
R5 is hydrogen or oxygen or is joined to R to form the partial structure:
Figure imgf000191_0002
and the bond represented by ____; is: 1 ) a double bond when p is zero or when p is 1 and R5 is oxygen, or
2) a single bond when R5 is hydrogen or R5 is joined to R2 to form the partial structure:
Figure imgf000192_0001
20. The selective Ij s compound of Claim 19 wherein
A is benzo either unsubstituted or substimted with -NH2, -NHSθ2(Cl-3 alkyl), Cl-3 alkyl or Cl-3 alkoxy;
X and Y are O,
Z is
1 ) Cl-6 alkylene, either straight or branch chain and either unsubstituted or substituted with phenyl or spiropiperidine,
2) C2-4 alkenylene, either straight or branch chain,
3) -(CH2)m- -(CH2)n- wherein m and n are independently 0, 1, 2, 3 or 4 and W is -0-, -S- or -NH,
4) 4-(5-methylisoxazole-3-yl),
5) C3-6 cycloalkylene, or
6) single bond;
Rl is
1 ) phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02, b) -CI, Br, F, or I, c) -CF3,
Figure imgf000193_0001
4) mono- or bicyclic heterocyclyl of 5 to 10 members one or two of which are sulfur, nitrogen or oxygen, the remaining being carbon, such as 2-thienyl, 2-furanyl, 2-indolyl, 2- quinoxolinyl, or 2-(2,3-dihydro benzofuranyl)
5) methyl, or
6) indan-5-yl;
R2 i IS
1 ) phenyl, either unsubstituted or substituted with Cl-3 alkoxy or 4,4-dimethyloxazolin-2-yl,
2) Cl-4 alkyl, either straight or branched chain and either unsubstituted or substituted with -3 alkoxy or
Cl -3 alkoxy-Ci-3 alkoxy,
3) C5-7 cycloalkyl,
4) 2- or 3-furyl,
5) l-methylpiperidin-2-yl, or
6) if R is phenyl, the 2-position of the phenyl can be joined to the 4-position nitrogen of the diazepine ring through a carbonyl group and the double bond between the 4-nitrogen and the 5-carbon becomes a single bond;
R3 IS
1 ) hydrogen or
2) Cl -6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3, or
3) -CF3; R is
1 ) hydrogen,
2) Cl-6 alkyl, the chain of carbon atoms of which can be interrupted by one or two non-adjacent oxygen atoms and which is either unsubstituted or substituted with Cl-3 alkoxy carbonyl, -OH or
Figure imgf000194_0001
3) tetrazol-5-yl; and
R5 is hydrogen or oxygen or is joined to R to form the partial structure:
Figure imgf000194_0002
the bond represented by ___^ is:
1) a double bond when p is zero or when p is 1 and R5 is oxygen, or
2) a single bond when R^ is hydrogen or R^ is joined to R to form the partial structure:
Figure imgf000194_0003
including individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt.
21. The selective Ij^s compound of Claim 20 wherein A is benzo either unsubstituted or substituted with -NH2, -NHS02 (Cl-3 alkyl), Cl-3 alkyl or Cl -3 alkoxy;
X and Y are O,
Z is
1 ) Cl -6 alkylane, either straight or branch chain and either unsubstituted or substituted with phenyl or spiropiperidine,
2) C2-4 alkenylene, either straight or branch chain,
3) -(CH2)m-W-(CH2)n- wherein m and n are independently 0, 1, 2, 3 or 4 and W is -O-, -S- or -NH,
4) 4-(5-methylisoxazole-3-yl),
5) C3-6 cycloalkylene, or
6) single bond;
Ri is phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02, b) -CI, Br, F, or I, c) -CF3, d) -Cl-3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy,
R2 is phenyl, either unsubstituted or substimted with Cl-3 alkoxy or 4,4-dimethyloxazolin-2-yl,
R3 is -CF3 or Cl-6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3;
R4 and R^ are hydrogen; including individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt.
22. The selective iKscompound of Claim 21 wherein A is unsubstituted benzo;
Rl is phenyl, either unsubstituted or substituted with one or two substituents selected from a) -Nθ2, b) -CI, Br, F, or I, c) -CF3, d) -Cl-3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy,
R2 is phenyl, either unsubstituted or substituted with Cl-3 alkoxy or 4,4-dimethyloxazolin-2-yl,
R3 is -CF3 or Cl-6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3;
R4 and R^ are hydrogen;
including individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt.
23. The selective Ij^s compound of Claim 22 which is
(-)-2-[2,4-Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5- phenyl-l-(2,2,2-trifluoroethyl)-lH-benzo[e][l,4]diazepin-3-yl]acetamide
Figure imgf000197_0001
24. The selective Ij s compound of Claim 22 which is 3,4-Dichloro-N-[2,3-dihydro-5-(2-fluorophenyl)-2-oxo-l -(2,2,2- trifluoroethyl)- 1 H-benzo[e] [ 1 ,4]diazepin-3-yl]benzamide
Figure imgf000197_0002
25. The selective Ij^s compound of Claim 22 which is (-)-2-(2,4-Dichlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5-phenyl-l- (2,2,2-trifluoroethyl)-lH-benzo[e][l,4]diazepin-3-yl]acetamide
Figure imgf000197_0003
26. The beta-adrenergic receptor blocking agent of Claim 19 selected from the group consisting of timolol, sotalol, esmolol, cateolol, propranolol, betaxolol, penbutolol, metoprolol, acebutolol, atenolol, metoprolol, pindolol, or bisoprolol.
PCT/US1997/011131 1996-06-28 1997-06-25 Pharmaceutical preparation WO1998000405A1 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
JP10504289A JP2000510155A (en) 1996-06-28 1997-06-25 Pharmaceutical preparations
EP97931437A EP0907644A4 (en) 1996-06-28 1997-06-25 Pharmaceutical preparation
CA002257948A CA2257948A1 (en) 1996-06-28 1997-06-25 Antiarrhythmic combinations of selective iks antagonists with beta-adenergic blocking agents
AU35066/97A AU722110B2 (en) 1996-06-28 1997-06-25 Pharmaceutical preparation

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US2074796P 1996-06-28 1996-06-28
US60/020,747 1996-06-28
GBGB9617894.2A GB9617894D0 (en) 1996-08-28 1996-08-28 Pharmaceutical preparation
GB9617894.2 1996-08-28

Publications (1)

Publication Number Publication Date
WO1998000405A1 true WO1998000405A1 (en) 1998-01-08

Family

ID=26309928

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1997/011131 WO1998000405A1 (en) 1996-06-28 1997-06-25 Pharmaceutical preparation

Country Status (5)

Country Link
EP (1) EP0907644A4 (en)
JP (1) JP2000510155A (en)
AU (1) AU722110B2 (en)
CA (1) CA2257948A1 (en)
WO (1) WO1998000405A1 (en)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000069836A1 (en) * 1999-05-14 2000-11-23 Glaxo Group Limited Short-acting benzodiazepines
US6509331B1 (en) 1998-06-22 2003-01-21 Elan Pharmaceuticals, Inc. Deoxyamino acid compounds, pharmaceutical compositions comprising same, and methods for inhibiting β-amyloid peptide release and/or its synthesis by use of such compounds
US6528505B1 (en) 1998-06-22 2003-03-04 Elan Pharmaceuticals, Inc. Cyclic amino acid compounds pharmaceutical compositions comprising same and methods for inhibiting β-amyloid peptide release and/or its synthesis by use of such compounds
US6683075B1 (en) 1996-12-23 2004-01-27 Athena Neurosciences, Inc. Cycloalkyl, lactam, lactone and related compounds, pharmaceutical compositions comprising same, and methods for inhibiting β-amyloid peptide release and/or its synthesis by use
US6774125B2 (en) 1998-06-22 2004-08-10 Elan Pharmaceuticals, Inc. Deoxyamino acid compounds, pharmaceutical compositions comprising same, and methods for inhibiting β-amyloid peptide release and/or its synthesis by use of such compounds
US6838455B2 (en) 1998-06-22 2005-01-04 Athena Neurosciences, Inc. Deoxyamino acid compounds, pharmaceutical compositions comprising same, and methods for inhibiting beta-amyloid peptide release and/or its synthesis by use of such compounds
US6906056B2 (en) 1998-06-22 2005-06-14 Elan Pharmaceuticals, Inc. Cycloalkyl, lactam, lactone and related compounds, pharmaceutical compositions comprising same, and methods for inhibiting β-amyloid peptide release and/or its synthesis by use of such compounds
US6958330B1 (en) 1998-06-22 2005-10-25 Elan Pharmaceuticals, Inc. Polycyclic α-amino-ε-caprolactams and related compounds
US7160880B1 (en) 1999-05-14 2007-01-09 Cenes Limited Short-acting benzodiazepines
US10195210B2 (en) 2010-11-08 2019-02-05 Paion Uk Ltd. Dosing regimen for sedation with CNS 7056 (Remimazolam)
US10414749B2 (en) 2013-03-04 2019-09-17 Paion Uk Limited Process for preparing 3-[(S)-7-bromo-2-((2-oxopropyl)amino)-5-pyridin-2-yl-3H-1,4-benzodiazepin-3-yl]propionic acid methyl ester
US10472365B2 (en) 2006-07-10 2019-11-12 Paion Uk Limited Short-acting benzodiazepine salts and their polymorphic forms

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1494685B1 (en) * 2002-04-18 2008-12-31 Cv Therapeutics, Inc. Method of treating arrhythmias comprising administration of an a1 adenosine agonist with a beta blocker
GB201613942D0 (en) * 2016-08-15 2016-09-28 Univ Of Durham The An antimicrobial compound

Citations (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0167919A2 (en) * 1984-06-26 1986-01-15 Merck & Co. Inc. Benzodiazepine derivatives and pharmaceutical compositions containing them
US4628084A (en) * 1986-01-02 1986-12-09 Merck & Co., Inc. Process for 3-acylamino benzodiazepines
US4820834A (en) * 1984-06-26 1989-04-11 Merck & Co., Inc. Benzodiazepine analogs
US4970207A (en) * 1988-07-07 1990-11-13 Fujisawa Pharmaceutical Company, Ltd. Benzodiazepine derivatives
US4992437A (en) * 1987-12-22 1991-02-12 Yoshitomi Pharmaceutical Industries, Ltd. Thienodiazepine compounds and their pharmaceutical use
US4994258A (en) * 1990-03-05 1991-02-19 Merck & Co., Inc. Gamma emitting, CCK-A antagonists for pancreatic imaging
US5153191A (en) * 1991-08-20 1992-10-06 Warner-Lambert Company Cholecystokinin antagonists useful for treating depression
US5220017A (en) * 1991-04-10 1993-06-15 Merck & Co., Inc. Cholecystokinin antagonists
US5324726A (en) * 1989-12-18 1994-06-28 Merck & Co., Inc. Benzodiazepine analogs
US5360802A (en) * 1992-05-11 1994-11-01 Merck Sharpe & Dohme Ltd. Benzodiazepine derivatives, compositions containing them and their use in therapy
WO1995014694A1 (en) * 1993-11-22 1995-06-01 Merck & Co., Inc. Benzodiazepines
US5426185A (en) * 1993-11-22 1995-06-20 Merck & Co., Inc. Antiarrhythmic benzodiazepines
US5428031A (en) * 1991-12-03 1995-06-27 Merck & Co., Inc. Methods of treating cardiac arrhythmia
US5439906A (en) * 1993-11-22 1995-08-08 Merck & Co., Inc. Antiarrhythmic benzodiazepines
US5439905A (en) * 1991-09-30 1995-08-08 Yoshitomi Pharmaceutical Industries, Ltd. Thienodiazepine compounds and their use
US5451582A (en) * 1992-02-21 1995-09-19 Merck Sharp & Dohme Ltd. Benzodiazepine derivatives, compositions containing them and their use in therapy
WO1996005827A1 (en) * 1994-08-18 1996-02-29 Merck & Co., Inc. N-2,3-dihydro-1-(2-propyl)-2-oxo-5-phenyl-1h-1,4-benzodiazepines
WO1996005839A1 (en) * 1994-08-18 1996-02-29 Merck & Co., Inc. 2,3-dihydro-1-(2,2,2-trifluoroethyl)-2-oxo-5-phenyl-1h-1,4-benzodiazepines
US5521175A (en) * 1992-07-29 1996-05-28 Merck Sharp & Dohme Ltd. Benzodiazepine derivatives
US5556969A (en) * 1994-12-07 1996-09-17 Merck Sharp & Dohme Ltd. Benzodiazepine derivatives
US5602125A (en) * 1992-08-28 1997-02-11 Merck, Sharp & Dohme, Ltd. Thienodiazepine derivatives as cholecystokinin and gastrin antagonists

Patent Citations (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0167919A2 (en) * 1984-06-26 1986-01-15 Merck & Co. Inc. Benzodiazepine derivatives and pharmaceutical compositions containing them
US4820834A (en) * 1984-06-26 1989-04-11 Merck & Co., Inc. Benzodiazepine analogs
US4628084A (en) * 1986-01-02 1986-12-09 Merck & Co., Inc. Process for 3-acylamino benzodiazepines
US4992437A (en) * 1987-12-22 1991-02-12 Yoshitomi Pharmaceutical Industries, Ltd. Thienodiazepine compounds and their pharmaceutical use
US4970207A (en) * 1988-07-07 1990-11-13 Fujisawa Pharmaceutical Company, Ltd. Benzodiazepine derivatives
US5324726A (en) * 1989-12-18 1994-06-28 Merck & Co., Inc. Benzodiazepine analogs
US4994258A (en) * 1990-03-05 1991-02-19 Merck & Co., Inc. Gamma emitting, CCK-A antagonists for pancreatic imaging
US5220017A (en) * 1991-04-10 1993-06-15 Merck & Co., Inc. Cholecystokinin antagonists
US5153191A (en) * 1991-08-20 1992-10-06 Warner-Lambert Company Cholecystokinin antagonists useful for treating depression
US5439905A (en) * 1991-09-30 1995-08-08 Yoshitomi Pharmaceutical Industries, Ltd. Thienodiazepine compounds and their use
US5428031A (en) * 1991-12-03 1995-06-27 Merck & Co., Inc. Methods of treating cardiac arrhythmia
US5597818A (en) * 1991-12-03 1997-01-28 Merck & Co., Inc. Methods of treating cardiac arrhythmia
US5451582A (en) * 1992-02-21 1995-09-19 Merck Sharp & Dohme Ltd. Benzodiazepine derivatives, compositions containing them and their use in therapy
US5360802A (en) * 1992-05-11 1994-11-01 Merck Sharpe & Dohme Ltd. Benzodiazepine derivatives, compositions containing them and their use in therapy
US5521175A (en) * 1992-07-29 1996-05-28 Merck Sharp & Dohme Ltd. Benzodiazepine derivatives
US5602125A (en) * 1992-08-28 1997-02-11 Merck, Sharp & Dohme, Ltd. Thienodiazepine derivatives as cholecystokinin and gastrin antagonists
US5595990A (en) * 1993-11-22 1997-01-21 Merck & Co., Inc. Antiarrhythmic benzodiazepines
US5426185A (en) * 1993-11-22 1995-06-20 Merck & Co., Inc. Antiarrhythmic benzodiazepines
US5439906A (en) * 1993-11-22 1995-08-08 Merck & Co., Inc. Antiarrhythmic benzodiazepines
WO1995014694A1 (en) * 1993-11-22 1995-06-01 Merck & Co., Inc. Benzodiazepines
WO1996005839A1 (en) * 1994-08-18 1996-02-29 Merck & Co., Inc. 2,3-dihydro-1-(2,2,2-trifluoroethyl)-2-oxo-5-phenyl-1h-1,4-benzodiazepines
WO1996005827A1 (en) * 1994-08-18 1996-02-29 Merck & Co., Inc. N-2,3-dihydro-1-(2-propyl)-2-oxo-5-phenyl-1h-1,4-benzodiazepines
US5633251A (en) * 1994-08-18 1997-05-27 Merck & Co., Inc. N-2,3-dihydro-1-(2-propyl)-2-oxo-5-phenyl-1h-1,4-benzodiazepines
US5658901A (en) * 1994-08-18 1997-08-19 Merck & Co., Inc. 2,3-dihydro-1-(2,2,2,-trifluoroethyl)-2-oxo-5-phenyl-1H-1,4-benzodiazepines
US5556969A (en) * 1994-12-07 1996-09-17 Merck Sharp & Dohme Ltd. Benzodiazepine derivatives

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP0907644A4 *

Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6683075B1 (en) 1996-12-23 2004-01-27 Athena Neurosciences, Inc. Cycloalkyl, lactam, lactone and related compounds, pharmaceutical compositions comprising same, and methods for inhibiting β-amyloid peptide release and/or its synthesis by use
US6958330B1 (en) 1998-06-22 2005-10-25 Elan Pharmaceuticals, Inc. Polycyclic α-amino-ε-caprolactams and related compounds
US6509331B1 (en) 1998-06-22 2003-01-21 Elan Pharmaceuticals, Inc. Deoxyamino acid compounds, pharmaceutical compositions comprising same, and methods for inhibiting β-amyloid peptide release and/or its synthesis by use of such compounds
US6696438B2 (en) 1998-06-22 2004-02-24 Elan Pharmaceuticals, Inc. Cyclic amino acid compounds, pharmaceutical compositions comprising same, and methods for inhibiting β-amyloid peptide release and/or its synthesis by use of such compounds
US6528505B1 (en) 1998-06-22 2003-03-04 Elan Pharmaceuticals, Inc. Cyclic amino acid compounds pharmaceutical compositions comprising same and methods for inhibiting β-amyloid peptide release and/or its synthesis by use of such compounds
US6774125B2 (en) 1998-06-22 2004-08-10 Elan Pharmaceuticals, Inc. Deoxyamino acid compounds, pharmaceutical compositions comprising same, and methods for inhibiting β-amyloid peptide release and/or its synthesis by use of such compounds
US6838455B2 (en) 1998-06-22 2005-01-04 Athena Neurosciences, Inc. Deoxyamino acid compounds, pharmaceutical compositions comprising same, and methods for inhibiting beta-amyloid peptide release and/or its synthesis by use of such compounds
US6906056B2 (en) 1998-06-22 2005-06-14 Elan Pharmaceuticals, Inc. Cycloalkyl, lactam, lactone and related compounds, pharmaceutical compositions comprising same, and methods for inhibiting β-amyloid peptide release and/or its synthesis by use of such compounds
WO2000069836A1 (en) * 1999-05-14 2000-11-23 Glaxo Group Limited Short-acting benzodiazepines
US7473689B2 (en) 1999-05-14 2009-01-06 Cenes Limited Short-acting benzodiazepines
US7435730B2 (en) 1999-05-14 2008-10-14 Cenes Limited Short-acting benzodiazepines
US7485635B2 (en) 1999-05-14 2009-02-03 Cenes Limited Short-acting benzodiazepines
US7528127B2 (en) 1999-05-14 2009-05-05 Cenes Limited Short-acting benzodiazepines
US7160880B1 (en) 1999-05-14 2007-01-09 Cenes Limited Short-acting benzodiazepines
US10961250B2 (en) 2006-07-10 2021-03-30 Paion Uk Limited Short-acting benzodiazepine salts and their polymorphic forms
US10472365B2 (en) 2006-07-10 2019-11-12 Paion Uk Limited Short-acting benzodiazepine salts and their polymorphic forms
US10195210B2 (en) 2010-11-08 2019-02-05 Paion Uk Ltd. Dosing regimen for sedation with CNS 7056 (Remimazolam)
US10342800B2 (en) 2010-11-08 2019-07-09 Paion Uk Ltd. Dosing regimen for sedation with CNS 7056 (Remimazolam)
US10414749B2 (en) 2013-03-04 2019-09-17 Paion Uk Limited Process for preparing 3-[(S)-7-bromo-2-((2-oxopropyl)amino)-5-pyridin-2-yl-3H-1,4-benzodiazepin-3-yl]propionic acid methyl ester

Also Published As

Publication number Publication date
CA2257948A1 (en) 1998-01-08
EP0907644A1 (en) 1999-04-14
JP2000510155A (en) 2000-08-08
EP0907644A4 (en) 2003-02-05
AU3506697A (en) 1998-01-21
AU722110B2 (en) 2000-07-20

Similar Documents

Publication Publication Date Title
RU2155587C2 (en) Derivatives of diazepine, pharmaceutical composition and method of treatment of patients with arrythmia
AU692916B2 (en) N-2,3-dihydro-1-(2-propyl)-2-oxo-5-phenyl-1h-1,4- benzodiazepines
US5428031A (en) Methods of treating cardiac arrhythmia
AU692300B2 (en) 2,3-dihydro-1-(2,2,2-trifluoroethyl)-2-oxo-5-phenyl-1h-1, 4-benzodiazepines
AU722110B2 (en) Pharmaceutical preparation
AU704220B2 (en) Novel 5-cyclopropyl-1,4 benzodiazepine-2-ones
US5776930A (en) Pharmaceutical preparation
AU703665B2 (en) Novel N-(2,4-dioxo-2,3,4,5-tetrahydro-1H-1,5-benzodiazepin-3 yl)-3-amides
US5817658A (en) Method for treating Meniere&#39;s disease
WO1997049690A1 (en) A method for treating meniere&#39;s disease
NZ328938A (en) Arrhythmia treatment using benzodiazepine derivatives
HUT74740A (en) Antiarrhythmic benzodiazepines and pharmaceutical compositions containing them
JP2001520197A (en) Novel benzodiazepine derivatives effective as antiarrhythmic drugs

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AU AZ BA BB BG BR BY CA CN CU CZ EE GE HU IL IS JP KG KR KZ LC LK LR LT LV MD MG MK MN MX NO NZ PL RO RU SG SI SK SL TJ TM TR TT UA US UZ VN YU AM AZ BY KG KZ MD RU TJ TM

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH KE LS MW SD SZ UG ZW AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 1997931437

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2257948

Country of ref document: CA

Ref country code: CA

Ref document number: 2257948

Kind code of ref document: A

Format of ref document f/p: F

WWP Wipo information: published in national office

Ref document number: 1997931437

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1997931437

Country of ref document: EP