AU722110B2 - Pharmaceutical preparation - Google Patents

Description

WO 98/00405 PCT/US97/11131 -1-

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 betaadrenergic 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.

WO 98/00405 PCT/US97/11131 -2- 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 III 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 HI 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 II 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 incorporated 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 components of cardiac delayed rectifier K+ current: differential WO 98/00405 PCT/US97/11131 -3sensitivity to block by Class III 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 NaCI, 4KC1, 1.2 MgCl2, 10 HEPES, glucose: pH 7.2, temp. 35 0

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 IKI 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 "betablockers", 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 WO 98/00405 PCT/US97/11131 -4patients (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 betaadrenergic 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 coadministration, 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 coadministration, 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.

WO 98/00405 PCT/US97/11131 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 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, 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-adrenergic receptor blocking agent which reaches the receptor sites.

Examples of compounds which fit the definition of betaadrenergic 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: R3 I X Y A IN Z-R' N R

R

2

(R

5

I

or a pharmaceutically acceptable salt thereof, wherein A is 1) thieno, 2) pyrido, or 3) benzo either unsubstituted or substituted with -NH2 -NHSO2 (C -3 alkyl), C1-3 alkyl or Cl-3 alkoxy; WO 98/00405 WO 9800405PCTIUS97/11131 -6- X is 1) =0, 2) =S, 3) =N-NH2, 4) =N-OH or =H2; Y is 1) =0, 2) =N-CN or 3) =H2; Z i 1) C 1 -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 or -Nil, 4) 4-(5-methylisoxazole-3-yl), C3-6 cycloalkylene, or 6) single bond; p is 0Oorl1; RI is 1) phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02, b) -Cl, Br, F, or 1, c) -CF3, d) -Cl1-3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy, 2) C5-7 cycloalkyl, 3) SN- 00 2 t-BU, WO 98/00405 PCT/US97/11131 -7- 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, 2quinoxolinyl, or 2-(2,3-dihydro benzofuranyl) Cl-3 alkyl, or 6)

R

2 is 1) phenyl, either unsubstituted or substituted with C1-3 alkoxy or 4,4-dimeth-yloxazolin-2-yl, 2) C1-6 alkyl, either straight or branched chain, and either unsubstituted or substituted with C1-3 alkoxy or C -3 alkoxy-C1-3 alkoxy, 3) C5-7 cycloalkyl, 4) 2- or 3-furyl, 1-methylpiperidin-2-yl, or 6) if R 2 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

3 is 1) hydrogen or 2) C -3 alkyl either unsubstituted or substituted with -N(CH3)2, -OH, -CF3, or 3) -CF3;

R

4 is 1) hydrogen, 2) C1-6 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 Ci-3 alkoxycarbonyl, -OH or N0 2 or 3) WO 98/00405 PCT/US97/11131 -8-

R

5 is hydrogen or oxygen or is joined to R2 to form the partial structure:

N

SO; and the bond represented by s: 1) a double bond when p is zero or when p is 1 and R 5 is oxygen, or 2) a single bond when R 5 is hydrogen or R 5 is joined to R2 to form the partial structure:

N

7 0--

N

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 quarternary ammonium salts of the compounds of Formula I formed, 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 WO 98/00405 PCT/US97/11131 -9contain 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;

R

3 is methyl;

R

4 is hydrogen; and

R

2 is C1-6 alkyl.

Specific novel compounds representative of this embodiment are those of the following structure and specified in Table I: TABLE I

CH

3 N

N

-N H

R

2 WO 98/00405 PCT/US97/11131

R

1 R2 2,4-diCIPh -CH3 2,4-diCIPh 2,4-diCIPh -t-Bu 4-CF3Ph i-C3H7 cyclohexyl i-C3H7 2,4-diCiPh i-C3H7 Another embodiment of the compounds useful in the novel method of treatment of this invention is that wherein: A is X and Y are oxygen;

R

3 is methyl;

R

4 is hydrogen; and

R

2 is phenyl.

A class of novel compounds within this embodiment is that with structural formula: WO 98/00405 WO 9800405PCTIUS97/11131 -11

OH

3 0 *N Z- R'

H

wherein Z is C1 -6 alkylene or a bond and R I is phenyl, phenyl substituted with -Cl, -Br, or -CF3, orR is cyclohexyl.

Specific novel compounds representative those depicted in the following Table II: of this class are TABLE II -(CH2)2- -(CH2)2- -(CH2)2- -(CH2)2- -(CH2)2- -CH2- -(CH2)2- -(CH2)2- -(CH2)2- 2 ,4-diClPh 4-CiPh 2,4-diFPh 2-CIPh 4-CF3Ph 4-CF3Ph 3-CF3Ph 2-CF3Ph cyclohexyl cyclohexyl WO 98/00405 WO 9800405PCT/US97/11131 12 TABLE 11 (Cont'd) z -(CH2)3 -CH2- -(CH2)2- -CH2- -(CH2)2-

-(CHL)-

-(CH2)3 -(CH2)2- -(CH2)3 Another class of novel is that with structural formnula: cyRheI cyclohexyl Pyhey Ph 4-CNPh -ClPh1- Ph 3-CNPh 2-thienyl compounds within this embodiment o0 N Z- R'

H

wherein Z is C2-4 alkenylene and R I is phenyl or phenyl substituted with -Cl, -Br, -CF3, CI-3 alkyl, CI-3 alkoxy or methylenedioxy.

Specific novel compounds representative of this class are those depicted in the following Table MI: WO 98/00405 WO 9800405PCT/US97/11131 13 TABLE III Z R -CH=CH- 4-NO 2 Ph -CH=CH- 2,4-diCIPh -CH=CH- 3-CIPh -CH=CH- 2-CIPh -CH=CH- 2,4-diFPh -CH=CH- 2,6-diCiPh -CH=CH- 4-CF 3 Ph -CH=CH- 2-BrPh WO 98/00405 WO 9800405PCTIUS97/11131 14 TABLE III (Cont'd) z

-CH=CH-

-CH=CH-

-C=CH-

OH

3

-CH=CH-

-CH=CH-

-CH=CH-

4-I1 4-BrPh 4BPh Ph 3 ,4-di ClPh 4-CH 3 Ph 4-CH 3 OPh -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 -Nil-.

Compounds representative of this embodiment are those disclosed in the following Table IV.

WO 98/00405 WO 9800405PCTfUS97/11131 15 TABLE IV I 3 y Al N )'NH-R' N

H

benzo 3-OH 3 Ph benzo 2,4-diCIPh benzo 3-CH, 3 Ph Ph \71'

OH

Ph

-OH

3

VON

I

benzo -OH 2 Cyclohexyl benzo 3-CH 3 Ph n-C 3

H

7

-OH

3

-OH

3 0 0 0

=N-CN

0 benzo 5-incianyl 3-OH 3 Ph Ph

OH

-OH

3 Other specific compounds included within the broadest genus but not included in one of the embodiments previously described are as shown in Table V.

WO 98/00405 PCT/US97/11131 16- TABLE V

Y

N" Z-R 1

I

R

4 Representative of compounds wherein p is 1 is the compound of structural formula:

CH

3 I n r Representative of compounds wherein the bond between the 4 and 5 positions is a single bond is the compound of structural formula:

CH

3

N

I N C NH H

CH

3

CH

3 WO 98/00405 PCT/US97/11131 17- Representative of compounds wherein the bond represents a single bond and R 5 is joined to R 2 is the compound of structural formula:

CH

3

I

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.

CH

3 I x y A N

Z-R

1 N I

R

4 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: WO 98/00405 PCT/US97/11131 18

CF

3 N H (CH 2 )n y

R

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: WO 98/00405 PCT/US97/11131 19- R1and R 2 are independently 1) phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02, OH, b) -Cl, Br, F, or I, c) -CF3, d) -C1-3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy, and Z is 1) C1-6 alkyl, either straight or branched chain 2) substituted Cl-6 alkyl, either straight or branched chain, wherein the substituents are selected from F, OH, N02, 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 -Sor -NH, 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 I, 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 tachyarrhythmias.

Individually, these compounds are effective in treating WO 98/00405 PCT/US97/11131 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 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 components of cardiac delayed rectifier K+ current: differential sensitivity to block by Class HI 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 NaCI, 4KC1, 1.2 MgCl2, 10 HEPES, glucose: pH 7.2, temp. 35 0

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 IKI is measured as WO 98/00405 PCT/US97/11131 -21 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 HO Z-R' N I -NH2 N .N 1. (COCI) 2 2. Ph 2. Et 3 Nj or 2

-N

Et 3 N

N

EDC

Z-R

1 HOBT N H WO 98/00405 WO 9800405PCTIUS97/11131 22 -v Br SCHEME 2 2. Br~t"B

H

2

NR

1 \N 0 iI,~K~Br -s N H Ph N.

NHR

-N H WO 98/00405 WO 9800405PCTJUS97/1 1131 23 SCHEME 3 3

H

2 S0 4 ""1NH 2 -7 1 11" INH 2 N0 2 1. R1-Zl CI 2. TiCI 3 3. Chromatography 'N IZ-Rl C H 3

SO

2 Cl

H

2

N

0

CH

3

SO

2 HN

N

N

'N IZ-R'

H

WO 98/00405 WO 9800405PCT/US97/11131 -24- SCHEME 4

OCN

H H 61 13. kh SCHEME j

NH

2 1. tBUCOCI

HOI

EtOH- H 2 0 2. n-BuLi 3. PhCONMe 2 1. BOC-Gly, DCC 2. TFA 0 H ,NH 2

N

A

;0 Ph Pyridine K-

N

Ph Ac

N

N

N ay N WO 98/00405 WO 9800405PCTIUS97/11131 25 SCHEME 6 0 Ph)I

CN

STOH

HO4; Et 3

N

S

NH

2 0 Ph 2. H 2

NNH

2 3. AcOH WO 98/00405 WO 9800405PCT/US97/11131 26 SCHEME 7 NaH, Mel KOtBu 0 5 H,,0N0_

N-

A

NH

2 1.Ph

:NOH

H

2 Ra-Ni or SnCI 2 or

N.

WO 98/00405 WO 9800405PCT/US97/1 1131 27 SCHEME 8

R'NCO

or -N2R'-Z

COO!

or

R

1

-Z-CO

2

H

EDO, HORT 0 ~N R1 N H N a-- No

S

WO 98/00405 WO 9800405PCTIUS97/11131 28 SCHEME 9 03 1. Lawesson's R 3 ReagentS )HNCBZ

HNCBZ

N

N

P h P h R3 H, Me 1. HBr

NI

2. 0 -N H cI Ph /H

R

3 =Me

H

2

NNH

2 Ra-Ni

NH

2 WO 98/00405 WO 9800405PCTIUS97/11131 29 SCHEME

NH-

2 GLY Ethyl ester 0 1) KOtBu/lsoamylnitrite 2) ethyl isocyanate 3) hydrogenation NaH cl I WO 98/00405 WO 9800405PCTIUS97/11131 30 SCHEME 11 m INH 2

R

4 X XSNH 3

CH

3

CN

H

X Br, CI, OTs, OMs.

0

CH

2

CI

2 000 ,u00

N

WO 98/00405 WO 9800405PCTfUS97/11131 31 SCHEME 12 B H 2

NH

2 xs NaHCO 3

CH

3

CN

0 -CV AZ-Rl

CH

2

CL

2

R.T.

,N,(0H 2

)Q

H 0.F\

H

2

)Q

Y0 0 WO 98/00405 WO 9800405PCTIUS97/11131 32 SCHEME 13 (1300 2 0, THF NaH R-MgX, THF -780C 80-30% X Br, Cl 'Boc 1) HCI, EtOAc 2) 1iN NaOH

OH

3 K-OtBu, Toluene isoamylnitrite -780C

NOH

1) ethyl isocyanate, THF 2) 10% Pd/C MeOH or Na 2

S

2

O

4 /TH F/H 2 0

OH

3 EDO, HOBT, TEA R' -Z-OOOH NI-1290%

OH

3 0 N ZR 1

H

WO 98/00405 WO 9800405PCTfUS97/11131 33 SCHEME 14 DM F-DMA Cbz Toluene N H N- Cbz

H

CH

3 HBr/AcOH

NH

2

CH

3 EDC, HOBT, TEA

R-Z-COOH

CH

3

CH

3 SCHEME 0 N MCPBA

N-~

-NH

WO 98/00405 WO 9800405PCTIUS97/11131 -34- SCHEME 16

I-CH

2

CF

3

CS

2 00 3

CF

3

CF

3 KOtBu

THE

F.

3

N

3

NH

2

H

2 Pd/C EtOH D-Cbz-Phe-OH, EDO, HOBT NHCbz

H

2 Pd/C, EtOH WO 98/00405 WO 9800405PCT/US97/11131 35

CF

3

NH

2 Separate Diastereomers

CEI

(CF

3 D-Phe-H 1) PhNCS 2) T FA N- D-Phe-H

H

CF

3

NH

2 ""1NH 2

R-(CH

2 ),COCI or

R-(CH

2 )nCO 2 H, EDC, HOBT N 0 1, Jil NL(CH 2 )n-R N

H

X, Y C1, CF: 3 Br n=0,1, or 2 R=~II or I X WO 98/00405 WO 9800405PCTIUS97/11131 36

EXAMPLES

EXAMPLE 1

OH

3 \0 0

NH

(3R) 1 -methyl-2-oxo-5-phenyl- 1 H-i ,4-benzodiazep~in-3 -yll -3-p2henyl -2-prop~enamide 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-i ,3-dihydro- 1 -methyl-S -phenyl-2H- 1 ,4-benzodiazepin-2one Ore. Chem. 1987, 52, 3232-3 239) (531 mg, 2.0 mmol) and triethylamine (307 mL, 225 mg, 2.2 mmnol) in methylene chloride 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/Et2O (95:5) and the residue was triturated with Et2O. The solid was collected and dried in vacuo at 70 0 C to give 2,3-dihydro-l1-methyl-2-oxo-5-phenyl- IH-I ,4-benzodiazepin-3-yl] -3phenyl-2-propenamide as a colorless solid (170 mg, 21 m.p. 140- 142 0 C, Mc~D +86.70 173, CH2C12).

dH (CDCl3) 7.70-7.26 (16H, in), 6.63 (IH, d, J 15.6 Hz), 5.68 (1H, d, J 8.3 Hz), and 3.50 (3H, s).

Anal. Calcd. for C25H21N302.0.15 (C2H5)20: C, 75.63; H, 5.58; N, 10.33.

Found: C, 75.29; H, 5.57; N, 10.33%.

WO 98/00405 WO 9800405PCTIUS97/11131 37 Employing the procedure substantialIly as described above, but substituting an appropriate acid chloride for the (E)-.3-phenyl-2propenoyl chloride, the following compounds were prepared: EXAMPLE 2

CH

3 N

H

-Dihydro- I-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazep~in- 3-yllbenzamide m.p. 224-225'C, MXD +89.20 (c 0.141, CH2Cl2).

dH (CDCl3) 8.04 (1H, d, J 8.1 Hz), 7.96 (2H, d, J 6.8 Hz), 7.64-7.36 (I1OH, in), 7.27 (2H, t, J 7.6 Hz), 5.74 (1 H, d, J 7.8 Hz), and 3.51 (3H,

S).

Anal. Calcd. for C23H119N302.0.20H20: C, 74.06; H, 5.24; N, 11.26.

Found: C, 74.13; H, 5.12; N, 11. 169o.

EXAMPLE 3

OH

3 WO 98/00405 WO 9800405PCTIUS97/11131 38 First diastereolsomer to elute: (-)-N-[(3R)-2,3-Dihydro-lI-methyl-2-oxo-5-phenyl-l1H- 1,4benzo-diazepin-3-yll (trans-2-phenyl- 1 -cycloprop~ane)carboxamide m.p. 180-181 0 C, [a]D -155.8' c 0.434, CH2C12).

dH (CDC13) 7.62-7.09 (15H, in), 5.59 (1 H, d, J 8.1 Hz), 3.47 (3H, s), 2.52-2.45 (1H, in), 1.90-1.84 (1H, m),1.69-1.56 (1H, in), and 1.38-1.32 (1I in A-nal. Cal U for f.26oi23N3O2.0.25H2O: C, 75.43; H, 5.72; N, 10.15.

Found: C, 75.38; H, 5.64; N, 9.94%.

Second diastereolsoiner to elute: (+)-N-[(3R)-2,3-Dihydro-l1-methyl-2-oxo-5-phenyl- 1H- 1,4benzo-diazepin-3 -yll (trans-2-12henyl- 1 -cyclop~ropane~carboxamide in.p. 104-107C, MoD +328.211 (c 0.098, CH2C12).

dH (CDC13) 7.62-7.13 (15H, in), 5.60 (1 H, d, J 8.3 Hz), 3.48 (3H, s), 2.59-2.54 (1H, in), 1.93-1.87 (1H, m),1.62-1.56 (1H, in, overlaps with water), and 1.33-1.25 (1H, in).

Anal. Calcd. for C26H23N302.0.50H20.O.45PhCH3: C, 76.13; H, 5.95; N, 9.14.

Found: C, 76.10; H, 5.94; N, 9.17%.

EXAMPLE 4 WO 98/00405 WO 9800405PCT/US97/11131 39 [(3R)-2,3-Dihydro- I -methyl-2-oxo-5-phenyl- 1 H-i A,benzodiazepin-3 -vii-1 H-indole-2-carboxamide m.p. 167-177C, Ml]) +1130 (c 1.103, CH2C12).

dH (CDC13) 9.15 (1 H, br 8. 10 (1IH, d, J 9.0 Hz), 7.75-7.10 (14H, i) 5.75 (1H, d, J 9.0 Hz), and 3.50 (3H, s).

Anal. Calcd. for C25H20N402: C, 73.51; H, 4.94; N, 13.72.

Found: C, 73.3 1; H, 4.80; N, 13.62%.

EXAMPLE (+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5 -phenyl- 1 H-i ,4-benzodiazepin-3 -yllhep~tanamide m.p. 49-54 0 C, [cx]D +69.50 (c=1.000, MeOH).

Anal. Calcd. for C23H27N302.0.40H20: C, 71.81; H, 7.28; N, 10.92.

Found: C, 71.90; H, 7.09; N, 10.85%.

EXAMPLE 6 100

N

1

NN

H

WO 98/00405 WO 9800405PCTIUS97/11131 [(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H-i ,4-benzodiazepin-3-yllhexanamide [tx]D +72.60 (c=0.920, MeOH).

Anal. Calcd. for C22H25N302: C, 72.70; H, 6.93; N, 11.56.

Found: C, 72.44; H, 6.75; N, 11.25%.

EXAMPLE 7 100

H

(+)-N-[(3R)-2,3-Dihydro-l1-methyl-2-oxo-5-phenyl- 1H-i ,4-benzodiazepin-3 -yllpentanamide +68.20 (c=1.3 10, MeOH).

Anal. Caled. for C2 1 H23N302.0.25CHC13: C, 68.21; H, 6.26; N, 11.26.

Found: C, 68.2; H, 6.29; N, 11.17%.

WO 98/00405 PCT/US97/11131 -41 EXAMPLE 8

CH

3

\OO

N

(+)-N-[(3R)-2,3-Dihydro-l-methyl-2-oxo-5-phenyl-lH-1,4-benzodiazepin-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-1,4benzodiazepin-2-one 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 mL) and extracted with ethyl acetate (3 x 20 mL). The combined organic fractions were dried (Na2SO4) and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2Cl2/Et20 (95:5) and the residue was recrystallized from toluene/hexane to give dihydro-1 -methyl-2-oxo-5-phenyl-1 H-1,4-benzodiazepin-3-yl] -3phenylpropanamide as a colorless solid (380 mg, m.p. 179 0

C,

[a]D +100.40 (c 0.225, CH2C12).

dH (CDC13) 7.62-7.57 (2H, 7.47-7.21 (13H, 5.54 (1H, d, J 8.1 Hz), 3.47 (3H, 3.03 (2H, t, J 7.8 Hz), and 2.73-2.67 (2H, m).

Anal. Calcd. for C25H23N302.0.15H20: C, 75.04; H, 5.87; N, 10.50.

Found: C, 75.06; H, 5.78; N, 10.55%.

WO 98/00405 WO 9800405PCT/US97/1 1131 42 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 (3R) E-(-i)-N-[(3R)-2,3-Dihydro-l1-methyl-2-oxo-5 -phenyl- 1H-I ,4-benzodiazep~in-3-vll -3 .4-dichlorophenyl)-2-p2ropenamide m.p. 145-147 0 C, [cX]D +77.8O (c=0.126, CH2C12).

dH (CDCl3) 7.64-7.25 (14H, in), 6.61 (iR, d, J 15.6 Hz), 5.65 (1H, d, J Hz), and 3.50 (3H, s).

Anal. Calcd. for C25H1I9N302C12: C, 64.67; H, 4.12; N, 9.05.

Found: C, 64.57; H, 4.25; N, 9.01 EXAMPLE N0 2 (3R) WO 98/00405 WO 9800405PCTIUS97/11131 43 E-(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5 -phenyl- 1 H-i ,4-benzodiazepin-3-vll 3-(4-nitrophenyl)-2-prop~enamide m.p. 165-166 [cz]D +80.50 (c=0.126, CH2CI2).

dH (CDCl3) 8.26 (1 H, d, J 8.8 Hz), 7.74-7.28 (1311, in), 6.76 (1 H, d, J 15.6 Hz), 5.66 (1H, d, J 8.0 Hz), and 3.51 (3H, s).

Anal. Caled. for C25H 1 9N404: C, 68.17; H, 4.58; N, 12.72.

Found: C, 68.25; H, 4.65; N, 12.57%.

EXAMPLE 11I

OH

3

NH

ci E-(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5 -phenyl- 1 H-i ,4-benzodiazepin-3-yll -3 -(2..4-dichlorophenyl)-2-propenamide m.p. 137-139'C, [RX]D +66.00 (c=0.144, CH2CI2).

dH (CDCI3) 8.02 (1H, d, J 15.6 Hz), 7.73-7.26 (13H, in), 6.66 (1H, d, J 15.6 Hz), 5.8.1 (1H, d, J 8.8 Hz), and 3.53 (3H, s).

Anal. Calcd. for C25H1I9012N302: C, 64.67; H, 4.12; N, 9.05.

Found: C, 64.28; H, 4.24; N, 8.83%.

WO 98/00405 WO 9800405PCTIUS97/11131 44 EXAMPLE 12

CH

3 E-(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1H- 1,4benzodiazepin-3 -vi 3-(4-methylp~henyl)-2-p2ropenamide m.p. 133-135'C, [aCID +90.40 125, CH2C12).

dH (CDCI3) 7.68-7.19 (15H, in), 6.59 (1H, d, J 15.6 Hz), 5.70 (1H, d, J Hz), 3.50 (3H, and 2.38 (3H, s).

Anal. Calcd. for C26H23N302: C, 76.26; H, 5.66; N, 10.26.

Found: C, 75.93; H, 5.82; N, 10.10%.

EXAMPLE 13

OH

3 0

~CH

3 E-(+)-N-[(3R)-2,3-Dihydro-l1-methyl-2-oxo-5-phenyl-l1H-I ,4-benzodiazepin-3-vl 1-3 -(4-methoxyphenvl)-2-propenamide m.p. 129-133 0 C, [a]ID +89.90 (c 0.188, CH2C12).

WO 98/00405 WO 9800405PCTIUS97/11 131 45 dH (CDCl3) 7.65-7.24 (14H, in), 6.92 (1H, d, J 8.8 Hz), 6.50 d, J 15.6 Hz), 5.69 (1H, d, J 8.0 Hz), 3.84 (3H, and 3.50 (3H, s).

Anal. Calcd. for C26H23N303.O.30H20: C, 72.48; H, 5.52; N, 9.75.

Found: C, 72.75; H, 5.60; N, 9.36%.

EXAMPLE 14

OH

3 1 0 0

CI

-NH

ci (+)-N-[(3R)-2,3-Dihydro-l1-methyl-2-oxo-5-phenyl- 1H- 1,4-benzodiazepin-3 -vii-3 -(2,4-dichlorophenyl)p2ropanamide m.p. 92-95'C, [RX]D 90.50 (c 0.196, CH2Cl2).

dH (CDCI3) 7.62-7.15 (13H, in), 5.52 (1 H, d, J 8.1 Hz), 3.47 (3H, s), 3.10 (2H1, t, J 7.6 Hz), and 2.68 (2H1, dd, J 7.6, 2.8 Hz).

Anal. Calcd. for C25H-21C12N302.0.20H-20: C, 63.89; H, 4.59; N, 8.94.

Found: C, 63.86; H, 4.62; N, 8.87%.

WO 98/00405 WO 9800405PCTIUS97/11131 46 EXAMPLE

OH

3 0 N

H

E-(+)-N-[(3R)-2,3-Dihydro-lI-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazepin-3 -yl-3 -chlorophenyl)-2-prop~enamide m.p. 229-231IT, [kX]D +86.20 (c 0.225, CH2Cl2).

dH (CDCl3) 7.64-7.26 (15H, in), 6.62 (1 H, d, J 15.6 Hz), 5.66 (1 H, d, J 8.1 Hz), and 3.50 (3H, s).

Anal. Calcd. for C25H120C1N302: C, 69.85; H, 4.69; N, 9.77.

Found: C, 70.20; H, 4.8 3; N, 9.41 1%.

EXAMPLE 16

OH

3 E-(-i)-N-F(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5 -phenyl- 1 H-i ,4-benzodiazepin-3-yl] -3-(2-chlorophenyl)-2-propenamide m.p. 128-1311C, MD =D +61.70 (c 0.196, CH2C12).

WO 98/00405 WO 9800405PCT/US97/11131 47 dH (CDCl3) 8.06 (1H, d, J 15.6 Hz), 7.65-7.28 (14H, in), 6.62, (LH, d, J 15.6 Hz), 5.68 (1H, d, J 8.3 Hz), and 3.50 (3H, s).

Anal. Calcd. for C25H20C1N302.0.20H20: C, 69.27; H, 4.74; N, 9.69.

Found: C, 69.21; H, 4.68; N, 9.45%.

EXAMPLE 17

OH

3 0

F

NII

NN

H

N F E-(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- LH- 1 ,4-benzodiazepin-3 -vii-3-(2,4-difluorophenV) -2-propenamide m-p. 121-123'C, IRx]D +76.80 (c 0. 111, CH2C12).

dH (CDC13) 7.71 (1 H, d, J 15.9 Hz), 7.64-7.24 (11 H, in), 6.92-6.84 (2H, in), 6.69 (1 H, d, J 15.9 Hz), 5.67 (1 H, d, J 8.1 Hz), and 3.50 (3H, s).

Anal. Caled. for C25H 1 9F2N302.0. 1 0H20: C, 69.3 1; H, 4.47; N, 9.70.

Found: C, 69.28; H, 4.57; N, 9.3 1 WO 98/00405 WO 9800405PCT/US97/11131 -48- EXAMPLE 18 -Dihydro- 1 -methyl -2-oxo-5 -phenyl- 1 H-i ,4-benzodiazepin-3-yll -3-(4-chlorophenyl)propanamide M.P. 203-205 0 C, WI~D +99.2' (c 0.300, CH2C12).

dH (CDC13) 7.62-7.16 (14H, in), 5.52 (1 H, d, J 8.1 Hz), 3.47 (3H, s), 2.99 (2H, t, J 7.7 Hz), and 2.67 (2H, t, J 7.7 Hz).

Anal. Caled. for C25H22C1N302: C, 69.52; H, 5.13; N, 9.73.

Found: C, 69.50; H, 5.15; N, 9.72%.

EXAMPLE 19

CH

3 E-(+)-N-II(3R)-2,3-Dihydro-l1-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzo- M.P. 121-124'C, RX]D +69.00 (c 0.342, CH2CI2).

WO 98/00405 WO 9800405PCTIUS97/11131 49 dH (CDC13) 7.79 (11H, d, J 16.1 Hz), 7.64-7.15 (13H, in), 6.78 (1H, d, J 15.8 Hz), 5.69 (1 H, d, J 8.1 Hz), and 3.50 (3H, s).

Anal. Calcd. for C25H19C12N302.0.15PhCH3: C, 65.44; H, 4.23; N, 8.79.

Found: C, 65.40; H, 4.38; N, 8.85%.

EXAMPLE

CH

3 1 00 Nj N

H

CF 3 E-(+)-N-II(3R)-2,3-Dihydro-l1-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazepmi-3 -vii-3- [4-(trifluoromethyl)p2henyll -2-propenamide m.p. 133-137C, MxD +68.70 (c =0.115, CH2C12).

dH (CDC13) 7.72-7.25 (15H, in), 6.71 (1H, d, J 15.6 Hz), 5.67 (L11, d, J 8.1 Hz), and 3.51 (3H, s).

Anal. Calcd. for C26H20F3N302: C, 67.38; H, 4.35; N, 9.07.

Found: C, 67.38; H, 4.45; N, 8.95%.

WO 98/00405 WO 9800405PCTIUS97/11131 50 EXAMPLE 21 (+)-5-Chloro-N-[(3R)-2,3 -dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1,4benzodiazep~in-3-yllindole-2-carboxamide m.p. 160-164'C, [x]D +103.80 (c 0.160, CH2C12).

dH (CDC13) 9.71 (1W, br 8.13 (1 H, d, J 7.8 Hz), 7 68-7.09 (13H, in), 5.75 (1IH, d, J 7.8 Hz), and 3.53 (3H, s).

Anal. Calcd. for C25H 1 C1N402.0.25H20.0. 15PhCH3: C, 67.84; H, 4.49; N, 12.15.

Found: C, 67.80; H, 4.41; N, 12.07%.

EXAMPLE 22

OH

3 [(3R)-2,3-Dihydro-l1-methyl-2-oxo-5-phenyl- 1H-I ,4-benzo- .diazep~in-3-vl] -2,2-diphenyiethanamide m.p. 200-201'C, [CC]D +97.00 (c 0.168, CH2CI2).

WO 98/00405 WO 9800405PCTIUS97/1 1131 51 dH (CDCl3) 7.60-7.22 (20H, in), 5.58 (1 H, d, J 8.1 Hz), 5.08 (1 H,s) and 3.44 (3H, s).

Anal. Calcd. for C30H-25N3015PhCH3: C, 78.79; H, 5.55; N, 8.88.

Found: C, 78.81; H, 5.63; N, 9.07%.

EXAMPLE 23

OH

3 0

F

N-j

NH

F

[(3R)-2,3-Dihydro-lI-methyl-2-oxo-5-phenyl- 1H-i ,4-benzodiazep~in-3-yll -3 -(2.4-difluorophenyl)p2ropanamide m.p. 79-81'C, [a]D +92.90 (c 0.105, CH2C12).

dH (CDCl3) 7.62-7.56 (3H1, in), 7.50-7.19 (8H, in), 6.82-6.76 (2H, in), 5.52 (1H, d, J 8.1 Hz), 3.47 (3H, 3.01 (2H1, t, J 7.6 Hz), and 2.69 (2H4, in).

Anal. Calcd. for C25H21F2N302: C, 69.27; H, 4.88; N, 9.69.

Found: C, 68.96; H, 4.99; N, 9.47%.

WO 98/00405 WO 9800405PCTIUS97/11131 52 EXAMPLE 24

CH

3 1 0 ,3 -Dihydro-l1-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazepin-3 -vii-2-phenylethanamide m.p. 241-242'C +85.5' (c 0.159, CH2CI2).

dH (CDCl3) 7.59-7.55 (3H, in), 7.46-7.22 (12H, in), 5.51 (LH, d, J 8.1Hz), 3.72 and 3.44 (3H, s).

Anal. Calcd. for C24H2 1N302.0.55H20: C, 73.28; H, 5.66; N, 10.68.

Found: C, 73.25; H, 5.38; N, 10.47%.

EXAMPLE

-N

NH

(+)-N-[(3R)-2,3-Dihydro- 1-methyi-2-oxo-5 -phenyl- 1H-i ,4-benzodiazep~in-3-yll -3-(2-chlorophenyl)p2rop~anamide m.p. 158.5-159.5 0 C, MID +95.80 (c 0.224, CH2CI2).

WO 98/00405 WO 9800405PCT/US97/11131 53 dH (CDC13) 7.62-7.57 (3H, in), 7.47-7.16 (11 H, mn), 5.55 (IH, d, J 8.1 Hz), 3.47 (3H, 3.14 (2H, t, J 7.9 Hz), and 2.75-2.69 (2H, in).

Anal. Caled. for C25H22C1N302.O.15H20: C, 69.09; H, 5.17; N, 9.67.

Found: C, 69.05; H, 5.12; N, 9.63%.

EXAMPLE 26

CH

3

NH

N'N

[(3R)-2,3-Dihydro- I-methyl-2-oxo-5 -phenyl- IH-i ,4-benzodiazepin-3 -yl] -3 -[4-(trifluoroinethvl)p2henvllpropanamide m.p. 175-176 0 C, [Rx]D +86.50 (c 0.141, CH2Cl2).

dH (CDC13) 7.62-7.54 (5H, in), 7.47-7.22 (9H, in), 5.52 (1H, d, J 8.1 Hz), 3.47 (3H, mn), 3.08 (2H, t, J 7.6Hz), and 2.72 (2H, in).

Anal. Calcd. for C26H22F3N302.0.80H20: C, 65.08; H, 4.93; N, 8.76.

Found: C, 65.03; H, 4.63; N, 8.72%.

WO 98/00405 WO 9800405PCTIUS97/1 1131 54 EXAMPLE 27

CH

3

.CF

3 (+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H-i ,4-benzodiazepin-3-yll [4-(trifluoromethyl)phenvllethanamide M.P. 224-226 0 C, [a]ID +68.0' (c 0.153, CH2C12).

dH (CDC13) 7.63-7.55 (4H, in), 7.5 1-7.33 (8H, in), 7.26-7.23 (2H, in), 5.51 (1iH, d, J 8.1 Hz), 3.77 (2H, 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

CH

3 k0 0 N CF3 N

H

-Dihydro-l1-methyl-2-oxo-5-phen'yl-l1H-i ,4-benzodiazep~in-3 -yll r3 -(trifluoromethy1~pheny1]propanamide m.p. 135-136'C, [aCID +78.80 (c 0.134, CH2C12).

WO 98/00405 WO 9800405PCT/US97/11131 55 dH (CDC13) 7.62-7.56 (3H1, in), 7.49-7.22 (1 1H, in), 5.53 (lH, d, J 8.1 Hz), 3.47 (3H, 3.08 (2H, t, J 7.3 Hz), and 2.72 (2H1, in).

Anal. Calcd. for C26H22F3N302: C, 67.09; H, 4.76; N, 9.03.

Found: C, 67.03; H, 4.73; N, 9.13%.

EXAMPLE 29

OH

3

N

N

H

(+)-3-Cyclohexyl-N- [(3R)-2,3-dihydro- 1 -methyl-2-oxo-5 -phenyl- 1 H- -1 4-benzodiazepin-3-yllpropanamide m.p. 144.5-145.5 0 C, MXD +83. 10 (c 0. 116, CH2C12).

dH (CDC13) 7.62-7.56 (3H1, in), 7.46-7.21 (7H, in), 5.55 (1H, d, J 8.3 Hz), 3.48 (3H1, 2.41-2.36 (2H1, in), 1.77-1.58 (7H1, in), 1.31-1.16 (4H, mn), and 0.98-0.90 (2H, in).

Anal. Calcd. for C25H29N302: C, 74.4 1; H, 7.24; N, 10.4 1.

Found: C, 74.46; H, 7.27; N, 10.58%.

WO 98/00405 WO 9800405PCT/US97/11 131 56 EXAMPLE

OH

3 I ,0

OF

3 (+)-N-[(3R)-2,3-Dihydro- i-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazepin-3 -yll -3 -[2-(trifluoromethyl)phenyllpropanamide m.p. 110- 1 13'C, [Ml) +79.20 (c 0.376, CH2C12).

dH (CDCl3) 7.65-7.57 (4H, in), 7.50-7.22 (10OH, in), 5.55 (1H, d, J Hz), 3.47 (3H, 3.20 (2H, t, J 7.9 Hz), and 2.70 (2H, dt, J 7.9, 3.3 Hz).

Anal. Calcd. for C26H22F3N302: C, 67.09; H, 4.76; N, 9.03.

Found: C, 66.97; H, 4.76; N, 8.93%.

EXAMPLE 31

OH

3

NO

-Dihydro- 1 -methyl -2-oxo-5 -phenyl- 1 H-i ,4-benzodiazep~in-3-yll -3 -(4-cyanophenyl')propanainide m.p. 81-85'C, [a]ID +91 1.00 (c 0. 111, CH2C12).

WO 98/00405 WO 9800405PCTIUS97/11131 57 dH (CDCl3) 7.64-7.55 (4H, in), 7.48-7.16 (10H, in), 5.50 (1H, d, J 8.3 Hz), 3.47 (3H, 3.08 (2H, t, J 7.6 Hz), and 2.74-2.69 (2H, in).

Anal. Calcd. for C26H22N4020.60H20.O.5OPhCH3: C, 73.93; H, 5.62; N, 11.69.

Found: C, 73.98; H, 5.61; N, 11.71%.

EXAMPLE 32

CH

3

NH

-Dihydro-l1-methyl-2-oxo-5 -phenyl- 1H-1I,4-benzodiazepin-3-yll-3-(3-chlorop~henyl)p2ropanamide m.p. 157-159'C, [cx]D +90.70 (c 0.134, C112C02).

dH (CDC13) 7.62-7.57 (3H, in), 7.47-7.12 (11IH, in), 5.53 (1H, d, J 8.1 Hz), 3.47 (3H, 3.00 (2H, t, J 7.3 Hz), and 2.7 1-2.66 (2H, in).

Anal. Calcd. for C25H22C1N302.0.5 5H20: C, 67.96; H, 5.27; N, 9.5 1.

Found: C, 67.99; H, 5.18; N, 9.26%.

WO 98/00405 WO 9800405PCTIUS97/11131 58 EXAMPLE 33

CH

3 0 Br

,N

H

E-(+)-N-[(3R)-2,3-Dihydro- 1-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazepin-3-vll -3 -(2-bromophenyl)-2-propenamide m.p. 113-1 16'C, MxD +44.20 (c =0.113, CH2CI2).

dH (CDC13) 8.03 (11H, d, J 15.6 Hz), 7.64-7.16 (14H, in), 6.57 (1H, d, J 15.6 Hz), 5.68 (1H, d, J 8.1 Hz), and 3.50 s).

Anal. Calcd. for C25H2OBrN3O2.O.60H20.0.3OPhCH3: C, 63.48; H, 4.58; N, 8.19.

Found: C, 63.49; H, 4.38; N, 8.19%.

EXAMPLE 34

OH

3 I. q 0 E-(+)-N-[(3R)-2,3-Dihydro-l1-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazepin-3 -bromophenyl)-2-p2rop~enamide m.p. 221-223 d 0 C, LOXID +65.50 (c =0.206, CH2Cl2).

WO 98/00405 WO 9800405PCTJUS97/1 1131 59 dH (CDCl3) 7.69 (1 H, br 7.64-7.57 (4H, in), 7.5 1-7.37 (6H1, in), 7.29-7.19 (4H, in), 6.62 (1WH d, J 15.6 Hz), 5.66 (11-1, d, J 8.1 Hz), and 3.50 (3H, s).

Anal. Calcd. for C25H2OBrN3O2.O.35H200.2OPhCH3: C, 63.54; H, 4.46; N, 8.42.

Found: C, 63.50; H, 4.39; N, 8.42%7.

EXAMPLE

CH

3 0 N

H

E-(+)-N-[(3R)-2,3-Dihydro-l1-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazepin-3-yll -3-(4-iodophenyl)-2-propenamide m.p. 137-140'C, [CC]D +67.90 (c 0.268, CH2C12).

dH (CDCl3) 7.75-7.72 (2H, in), 7.64-7.36 (8H, in), 7.29-7.16 (5H, in), 6.63 (1H, d, J 15.6 Hz), 5.66 (1H1, d, J 8.1 Hz), and 3.50 (3H, in).

Anal. Calcd. for C25H201N302.0.3OPhCH3: C, 59.29; H, 4.06; N, 7.65.

Found: C, 59.29; H, 3.90; N, 7.40%.

EXAMPLE 36 WO 98/00405 WO 9800405PCTIUS97/11131 1H 3 0

N'

E-(+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5 -phenyl- 1 H-i ,4-benzodi azepin-3 -vi]-3 -(4-bromophenyl)-2-p2rop~enamide m.p. 121-124 0 C, [o]ID +75.60 (c 0.201, CH2Cl2).

dH (CDCl3) 7.64-7.57 (3H, in), 7.55-7.35 (11 H, in), 7.28-7.24 (1 H, mn), 6.62 (1 H, d, J 15.6 Hz), 5.66 (1 H, d, J 8.1 Hz), and 3.50 (3H, s).

Anal. Calcd. for C25H20BrN3O2: C, 63.30; H, 4.25; N, 8.86.

Found: C, 63.50; H, 4.20; N, 8.78%.

EXAMPLE 37

OH

3 1 0 -N

H

-Dihydro-l1-methyl-2-oxo-5-phenyl- 1H-i ,4-benzodiazepin-3-yll -4-phenyibutanamide m.p. 65-74 0 C, [a]D +77.40 (c 0. 155, CH2C12).

dH (CDC13) 7.62-7.56 (3H, in), 7.46-7.19 (12H1, in), 5.55 (1H, d, J 8.1 Hz), 3.47 (3H, 2.71 (2H, t, J 7.6 Hz), 2.42-2.37 (2H, mn), and 2.09- 2.01 (2H1, in).

WO 98/00405 WO 9800405PCTIUS97/11131 61 Anal. Calcd. for C26H25N302.O.30H20: C, 74.91; H, 6.19; N, 10.08.

Found: C, 74.93; H, 6.05; N, 10.07%.

EXAMiPLE 38 [(3R)-2,3-Dihydro-l1-methyl-2-oxo-5-phenyl- 1H-i ,4-benzodiazep~in-3 -yll -5 -methyl -3 -p2henyli sox azol e-4 -carboxamide m.p. 123-126'C, [da1D 122.00 (c 0. 199, CH2Cl2).

dH (CDC13) 7.79-7.76 (2H, in), 7.62-7.32 (11 H, in), 7.26-7.21 (2H, in), 5.61 (1H, d, J 7.9 Hz), 3.42 (3H1, and 2.76 (311, s).

Anal. Calcd. for C27H22N403.0.40H20: C, 70.85; H, 5.02; N, 12.24.

Found: C, 70.84; H, 4.91; N, 11.92%.

EXAMPLE 39

OH

3 WO 98/00405 WO 9800405PCTIUS97/11131 -62- [(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H-i ,4-benzodiazepin- 3-vl -cyanophenyl)propanamide m.p. 110- 1 12'C, D +84.20 (c 0.202, CH2Cl2).

dH (CDCI3) 7.63-7.22 (1411, in), 5.51 (1H, d, J 8.1 Hiz), 3.47 (3H, s), 3.06 (2H, t, J 7.8 Hz), and 2.74-2.68 (2H4, in).

Anal. Calcd. for C26H22N402.0.50H20: C, 72.37; H, 5.37; N, 12.98.

Found:e C, 72. 5 2; H, 5. 12; N, 12. 5 9%.

EXAMPLE N- 0 Nj

H

(+)-N-[(3R)-2,3-Dihydro-l1-methyl-2-oxo-5 -phenyl- 1H- 1,4-benzodiazepin-3-yll cyclohexanethanamide m.p. 144-146'C, La]ID +72.1 0 (c=1.000, MeOH).

Anal. Calcd. for C24H27N302.0.20H20: C, 73.33; H, 7.03; N, 10.69.

Found: C, 73.27; H, 7.02; N, 10.76%.

WO 98/00405 WO 9800405PCTIUS97/1 1131 63 EXAMPLE 41 (+)-4-Cyclohexyl-N-[(3R)-2,3 -dihydro- 1 -methyl-2-oxo-5-phenyl- I H- 1 .4-benzodiazepin-3-yllbutanamide [a]ID +57.70 (c=0.440, MeOH).

Anal. Calcd. for C26H31N302: C, 74.79; H, 7.48; N, 10.06.

Found: C, 74.8;0 H, 7.78; N, 10.05%.

EXAMPLE 42 (+)-N-[(3R)-2,3-Dihydro-l1-methyl-2-oxo-5-phenyl- 1H-I ,4-benzodiazep~in-3-yll -4-methylpentanamide m.p. 123-125'C, [CC]D +66.80 (c=0.500, MeOH).

Anal. Calcd. for C22H25N3 02.O.45H20: C, 71.12; H, 7.03; N, 11.3 1.

Found: C, 71.08; H, 6.81; N, 11.42%.

WO 98/00405 WO 9800405PCTIUS97/11131 -64- EXAMPLE 43

H

3 C 0

N~

4 H 0 N H

(R)

-Dihydro- 1-methyl-2-oxo-5-phenyl- 1H-i ,4-benzodiazepin-3 -yll -2.3 -dihydrobenzofuran-2- carboxamide Dilsopropylethylamine (0.3 mL, 223 mg, 1.72 minol) was added to a stirred, cooled (0 solution of 3(R)-amino-i ,3-dihydro- 1methyl -5-phenyl-2H- 1,4-benzodiazepin-2-one Org. Chem. 1987, 52, 3232-3239) (400 mg, 1.5 mmol), 2,3-dihydrobenzofuran-2-carboxylic acid (274 mg, 1.7 mimol), 1-(3-dimethylammnopropyl)-3-ethylcarbodiimide hydrochloride (583 mg, 3.0 mmnol), and 1-hydroxybenzotriazole (479 mg, 3.1 mmol) in DMF (4.5 mL). The mixture was stiffed at room temperature for 18 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 (MgS 04) and evaporated under reduced pressure. The residue was crystallized from 2-chloro-2-methylpropanelhexane to give -dihydro- 1- 1H-i ,4-benzodiazepin-3 -yl] -2,3-dihydrobenzofuran-2-carboxamide as a colorless solid (156 mg, m.p. 141- 180 0 C, [1X]D 127.1 0 (c=0.425, CHCl3).

dH (CDC13) (3:1 Mixture of diastereoisomers) 8.44 in), 7.65-6.91 (13H, in), 5.52 (1H, in), 5.28 (11H, in), and 3.70-3.40 (514, in).

Anal. Calcd. for C25H21N303.0.25 Hexane C, 73.50; H, 5.70; N, 9.71.

Found: C, 74.12; H, 5.57; N, 9.71 1%.

WO 98/00405 PCT/US97/11131 EXAMPLE 44 (+)-N-[(3R)-2,3-dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H- 1,4-benzodiazepin-3-yl]-1 -dimethylethoxycarbonyl)spiro(cyclohexan-4,4'piperidine)-1 -carboxamide Step A: Et02C C02Et

N

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 Immol) 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 (100g) 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 (Na2SO4) and the solvent evaporated to give diethyl 1-benzylpiperidine-4,4-diacetate (37.5 1g).

A small portion of this was purified by flash column chromatography.

WO 98/00405 PCT/US97/11131 -66- NMR (300 MHz, CDC13) d: 7.2-7.4 5H), 4.11 J=7.3Hz,4H), 3.50 2H), 2.56 4H), 2.4 4H), 1.7 4H), 1.24 J=7.3Hz, 6H).

Step B: HO OH

N

,Ph 1 -Benzvlpiperidine-4,4-diethanol A solution of the diester (12.2 g, 35 mmol) in ether mL) was added to a cooled (-30 0 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), 1M NaOH (4.4 mL) and water 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 1-benzylpiperidine-4,4-diethanol.

m.p. 75-78 C NMR (300 MHz, CDC13) d: 7.2-7.4 5H), 3.7 J 6.8 Hz, 4H), 3.52 2H), 2.7 (brs, 2H), 2.43 4H), 1.66 J 6.8 Hz, 4H), 4H).

Step C:

BOC

WO 98/00405 PCT/US97/11131 -67- 1 -t-Butoxycarbonylpiperidine-4.4-diethanol The benzylamine (2.07 g, 7.9 mmol) was dissolved in methanol (60 mL), BOC20 (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-butoxycarbonylpiperidine-4,4-diethanol (2.0 g).

NMR (300 MHz, CDC13) d: 3.7 4H), d 3.3 6H), 1.65 J 6.8 Hz, 4H), 1.41 9H).

Step D: MeO 2 SO ,OSO 2 Me

N

BOC

1-t-Butoxycarbonvlpiperidine-4,4-diethanol, bis(methanesulfonate) The diol (2.41 g, 8.9 mmol) was dissolved in dichloromethene (50 mL), the solution cooled to -20 0 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 NaHCO3 and brine, dried (MgSO4) and the solvent evaporated to afford 1-t-butoxycarbonylpiperidine-4,4-diethanol, bis(methanesulfonate) (3.2g).

NMR (300 MHz, CDCl3) d: 4.32 J 7.1 Hz, 4H), 3.4 4H), 3.04 6H), 1.89 J 7.1 Hz, 4H).

WO 98/00405 WO 9800405PCT/US97/11131 68 Step E EIOOC COOEt Diethyl 3 -t-butyloxycarbonyl-3-azaspiro [5 .5lundecane-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 mniol). The mixture was cooled to 0 0 C and the bis-mesylate 1 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 (2x1 50 mL). The extracts were dried (Na2SO4), concentrated to an oil, and chromatographed on silica to give 3.83 g (60% of diethyl 3-t-butyloxycarbonyl-3-azaspiro- .5]undecane-9,9-dicarboxylate.

lH NMR (CDCl3) d: 1.22 6H), 1.4 9H), 2.0 (in, 4H), 3.35 (in, 4.2 4H).

BOC

WO 98/00405 PCT/US97/11131 -69- 3-t-Butyloxycarbonyl-3-azaspiro[5.5]undecane-9-carboxylic acid To a solution of the diester 2 (3.69 g, 0.0093 m) in THF mL) was added IN LiOH (47 mL). The reaction was stirred for 3 days at 25 0 C, diluted with water (50 mL) and pH adjusted to 2.2 with KHSO4. The product was extracted into ethyl acetate (2x75 mL), dried (Na2SO4), and concentrated to a foam (3.5 The solid was melted in a flask at 140 0 C for 2 hours, cooled and the oil dissolved in THF mL), IN LiOH (10 mL) added and mixture stirred overnight at 30 0

C.

The reaction was concentrated to remove THF, diluted with water mL) and washed with diethyl ether (10 mL). The pH was adjusted to with KHSO4 and product extracted (3x50 mL) with ethyl acetate.

The extracts were dried (Na2SO4), filtered and concentrated to yield 3t-butyloxycarbonyl-3-azaspiro[5.5]undecane-9-carboxylic acid as a foam (2.48 g, 1H NMR (CDC13, partial) d: 1.45 9H), 3.4 4H).

Employing the procedure substantially as described in Example 43 but substituting an appropriate acid for the 2,3-dihydrobenzofuran-2-carboxylic acid, the following compounds were prepared: Step G:

CH

3 0 N N-C-0-C-CH 3 III I H 0

C

H

3 (+)-N-[(3R)-2,3-Dihydro- -methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl]-1'-(1,1 -dimethylethoxycarbonyl)spiro(cyclohexan-4,4'piperidine)- 1-carboxamide m.p. 135-138 0 C, [r]D +58.80 (C=0.925, CHC13).

WO 98/00405 WO 9800405PCT/US97/1 1131 70 dH (CDCl3) 7.61-7.23 (10H, in), 5.54 (1H, d, J 9.0 Hz), 3.47 (3H, 3.37 (411, in), 2.28 (111, in), and 1.81-1.18 (21H, s).

Anal. Calcd. for C32H40N404: C, 70.56; H, 7.40; N, 10.29.

Found: C, 70.2 1; H, 7.40; N, 10. 16 EXAMPLE

H

3 0 0

NH

N H 0

(R)

(+)-N-[(3R)-2,3-Dihydro-l1-methyl-2-oxo-5-phenyl- 1H-i ,4-benzodiazep~in-3-yll-3-(furan-2-yI )propanamide M.P. 115-1 18'C, MD~ +65.81 (c=0.800, CHC13).

dH (CDCl3) 7.62-7.26 (11 H, mn), 6.28 (1 H, dd, J 3.2, 2.0 Hz), 6.08 (1 H, dd, J 3.2, 0.7 Hz), 5.58 (1 H, d, J 8.1 Hz), 3.48 (3H, 3.04 (2H, t, J 7.6 Hz), and 2.75 (2H, mn).

Anal. Calcd. for C23H21N303.0.3Hexane: C, 72.07; H, 6.15; N, 10.17.

Found: C, 71.78; H, 6.30; N, 9.77%.

WO 98/00405 WO 9800405PCTIUS97/11131 -71 EXAMPLE 46 [(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H-i ,4-benzodiazepin-3-yll (2-thienvl)butanamide m.p. 170-180'C, 1ex]D +63.50 (c=1.000, MeOH).

Anal. Calcd. for C24H23N302S.0.95H20: C, 66.32; H, 5.77; N, 9.67.

Found: C, 66.32; H, 5.34; N, 9.40%.

EXAMPLE 47

"IN

-Dihydro- I-methyl-2-oxo-5-phenyl- 1H-i ,4-benzodiazep~in-3 -viicyclohexylcarboxamide m.p. 213-214'C, MaD +62.40 (c=1.000, MeOH).

Anal. Calcd. for C23H124N302: C, 73.77; H, 6.46; N, 11.22.

Found: C, 73.86; H, 6.81; N, 11.15%.

WO 98/00405 WO 9800405PCT[US97/11131 72 EXAMPLE 48 (3R)-2,3 -Dihydro-l1-methyl-2-oxo -5 -phenyl- 1H-i ,4-benzodiazep~in-3-yl] -3-(3,4-methylenedioxyphenyl) -2-prop~enamide m.p. 143-145'C, [oC]D +62.30 (c=0.960, MeOH).

Anal. Calcd. for C25H2 1 N304O. 1 0H20.O.2OEt2O: C, 69.78; H, 5.27; N, 9.46.

Found: C, 69.78; H, 4.98; N, 9.28%.

EXAMPLE 49 -dihydro- 1-methyl -2-oxo-5-phenyl- 1H-i ,4-benzodiazepin-3-vll -2-guinoxalinecarboxamide [x]D +85.80 (c=0.360, MeOH).

Anal. Calcd. for C25H1I9N502: C, 69.96; H, 4.90; N, 15.33.

WO 98/00405 WO 9800405PCTIUS97/11131 -73- Found: C, 69.95; H, 4.72; N, 15.25%.

EXAMPLE -Dihydro-2-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazepin-3-yl] -2-(p2henvlamino~acetamide SWe A:

CH

3 Nj N0 B r

-N

H

N- [(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazepin- 3-vl -2-bromoacetamide Bromoacetyl bromide (165 mL, 383 mg, 1.9 mmol) was added to an ice cooled solution of 3 -amino-i 1,3 -dihydro-1I -methyl -5 phenyl-2H- 1,4-benzodiazepin-2-one Org. Chem,. 1987, 52, 3232- 3239) (500 mg, 1.88 mmol) and triethylamine (264 mL, 192 mg, 1.9 minol) in methylene chloride (10 mE) and the mixture was stirred at room temperature for 1 h. The mixture was washed with water (3 x mL), dried (MgSO4) and the solvent was evaporated under reduced pressure to give N-[(3R)-2,3-dihydro-l1-methyl-2-oxo-5-phenyl- 1H- 1,4benzodiazepin-3-yl]-2-bromoacetamide as a colorless foam (760 mg, 100%).

dH (CDCl3) 8.24 (1H, d, J 7.8 Hz), 7.64-7.24 (9H, in), 5.48 (1H, d, J 7.8 Hz), 4.00 (214, in), and 3.50 (3H, s).

WO 98100405 WO 9800405PCTIUS97/11131 -74-

OH

3 HO0 -Dihydro- 1 -methyl-2-oxo-5-phenyl- 11H-i ,4-benzodiazep~in-3-vll -2-(p2henylamino)acetamide Aniline (297 mL, 304 mg, 3.26 mmnol) was added to a solution of N-[(3R)-2,3-dihydro- I-methyl-2-oxo-5-phenyl- 1H- 1,4benzodiazepin-3-yl]-2-bromoacetamide (600 mg, 1.55 mmol) in ethanol 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 R)-2,3 -dihydro-l1-methyl-2-oxo-5 phenyl- 1H-i ,4-benzodiazepin-3-yl] -2-(phenylamino)acetamide as a colorless solid (500 mg, 81 m.p. 245-246 0 C, [a]D 1190 (C=0.850, CHC13).

dH (CDCl3) 8.26 (1H, d, J 8.3 Hz), 7.63-7.20 (12H, in), 6.81 (1H1, t, J 7.3 Hz), 6.72 (2H, d, J 7.6 Hz), 5.56 (1H, d, J 8.3 Hz), 3.95 (211, d, J Hz), and 3.45 (3H1, s).

Anal. Calcd. for C24H22N402: 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-(trifluoromethylDaniline for the aniline, the following compounds were prepared: WO 98/00405 WO 9800405PCT/US97/1 1131 75 EXAMPLE 51

OH

3 [(3R)-2,3-Dihydro-l1-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazepin-3-vH -2-(2-chlorophenylamino)acetamide m.p. 222-224TC, [cC]D +1110 (c=0.973, CHC13).

dH (CDCl3) 8.15 (1H, d, J 8.3 Hz), 7.60-7.16 (12H, in), 6.71 (2H, in), 5.57 (11H, d, J 8.3 Hz), 4.01 (2H, d, J 2.7 Hz), and 3.45 (3H, s).

Anal. Calcd. for C24H121 CIN402: C, 66.59; H, 4.89; N, 12.94.

Found: C, 66.40; H, 4.94; N, 12.92%.

EXAMPLE 52

OH

3 0 4I.NN N

NH

CF

3 (+)-N-[(3R)-2,3-Dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H-i ,4-benzodiazep~in-3-yll r4-(trifluoromethvl)phenylamino] acetamide m.p. 218-219 0 C, [Ml) +91.9' (c 0.419, CHC13).

WO 98/00405 PCTIUS97/11131 76 dH (CDC13) 8.13 (111, d, J 9.0 Hz), 7.70-7.25 (12H, in), 6.72 (2H, d, J 8.7 Hz), 5.60 (1 H, d, J 9.0 Hz), 4.05 (2H, in), and 3.50 (3H1, s).

Anal. Calcd. for C25H121F3N402.0.7H-20: C, 62.68; H, 4.71; N, 11.69.

Found: C, 62.47; H, 4.32; N, 11.44%.

EXAMPLE 53

CH

3 o N0 NN N)

H

-Dihydro- 1-methyl-2-oxo-5-phenyl- 1H-i ,4-benzodiazepin-3-yll -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, dihydro- 1-methyl-2-oxo-5-phenyl- 1H- 1,4-benzodiazepmn-3-yl] -2-bromoacetamide (400 mg, 1.04 mmol) was added and the mixture was stirred at room temperature for 18 h. The mixture was washed with water (3 x mL), dried (MgSO4) 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 [(3R)-2,3-dihydro- 1 -methyl-2-oxo-5-phenyl-l1H-i ,4-benzodiazepin-3yl]-2-(phenoxy)acetamide as a colorless solid (112 mg, m.p. 126- 128 0 C, [MD +81.6 (C=0.692, CHCl3).

dH (CDCI3) 8.49 (1H, d, J 8.2 Hz), 7.64-7.0 1 (14H, in), 5.61 (1H, d, J 8.2 Hz), 4.65 (1H, d, 1 14.6 Hz), 4.58 (1H, d, 1 14.6 Hz), and 3.50 (3H, s).

WO 98/00405 WO 9800405PCTIUS97/11131 77 Anal. Calcd. for C24H121N303: 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-dichiorophenol, thiophenol or 2,4-dichiorothiophenol for the phenol, the following compounds were prepared: EXAMPLE,54

OH

3 0 CI N Nj~ 0

H

[(3R)-2,3-Dihydro-l1-methyl-2-oxo-5 -phenyl- 1H-i ,4-benzodiazepin-3 -vl -2-(2..4-dichlorophenoxv)acetamide m.p. 206 0 C. MaD +3 1.10' (c=0.289, CHCl3).

dH (CDC13) 8.75 (1 H, d, J 9.0 Hz), 7.65 -7.20 (11 H, in), 6.90 (1 H, d, J 8.7 Hz), 5.60 (1 H, d, J 9.0 Hz), 4.65 (2H, in), and 3.50 (3H, s).

Anal. Calcd. for C241 9C12N303.0.3H20: C, 60.85; H, 4.17; N, 8.87.

Found: C, 60.80; H, 4.04; N, 8.87%.

WO 98/00405 WO 9800405PCTIUS97/11131 78 EXAMPLE

OH

3 -Dihydro -1 -methyl-2-oxo-5 -phenyl- 1 H-i ,4-benzodiazep~in-3-yll -2-(phenylthio)acetamide [cWD +104.90 (c=0.3 16, CHC13).

dH (CDC13) 8.50 (1H, d, J 9.0 Hz), 7.60-7.20 (14H, in), 5.50 (111, d, J Hz), 3.75 (2H, in), and 3.45 (3H, s).

Anal. Calcd. for C24H21N302S: C, 69.37; H, 5. 10; N, 10. 11.

Found: C, 68.98; H, 5.06; N, 9.76%.

EXAMPLE 56

CH

3 0 H ci (+)-N-jI(3R)-2,3-Dihydro- 1-methyl-2-oxo-5-phenyl-I H-i ,4-benzodiazepin-3-vyl -2-(2.4-dichlorophenylthio~acetamide RQ]D +97.40 (c=0.286, CHC13).

WO 98/00405 WO 9800405PCTIUS97/11131 79 dH (CDCl3) 8.35 (1H, d, J 9.0 Hz), 7.70-7.20 (12H, in), 5.50 (1H, d, J Hz), 3.70 (2H, in), and 3.50 (3H, s).

Anal. Calcd. for C24H19C12N3028: C, 59.51; H, 3.95; N, 8.67.

Found: C, 59.32; H, 3.95; N, 8.65%.

EXAMPLE 57

OH

3 0

H

[(3R)-2,3-Dihydro- 1-methyl-2-oxo-5-phenyl- 1H-i ,4-benzodiazep~in-3-vll -3-(phenylamino)p2rop~anamide 3-Bromopropionyl chloride (2.01 mL, 3.428 g, 20 mmol) was added to an ice cooled solution of 3(R)-amino-1,3-dihydro-1methyl-5-phenyl-2H-1 ,4-benzodiazepin-2-one (J Org. Chemi. 1987, 52, 3232-3239) (5.0 g, 18.8 mmol) and triethylamine (2.79 mL, 2.02 mg, 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 inL), dried (MgSO4) 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-l1-methyl-2-oxo-5 -phenyl-l1H-I ,4-benzodiazepin-3yl]-3-(phenylamnino)propanamide as a colorless solid, in.p. 218-221'C, [oX]D +58.21 (c=0.585, CHCl3).

WO 98/00405 WO 9800405PCTIUS97/11131 dH (CDC13) 7.60-6.7 1 (16H, in), 5.54 (1 H, d, J 8.1 Hz), 3.54 (2H, t, J 6.1 Hz), 3.52 (3H, and 2.70 (2H, mn).

Anal. Caled. for C25H24N402.O.5EtOH: C, 71.70; H, 6.25; N, 12.87.

Found: C, 71.42; H, 5.98; N, 12.84%.

EXAMPLE 58

CH

3 0 C1.

N H H C -Dihydro-l1-methyl-2-oxo-5-phenyl- 1H-I ,4-benzodiazepin-3-yll -3 -(2..4-dichlorophenvl)urea 2,4-Dichiorophenylisocyanate (188 mg, 1.0 mmol) was added to a solution of 3(R)-amino-1,3-dihydro-1-methyl-5-phenyl-2H- 1 ,4-benzodiazepin-2-one Org. Chem. 1987, 52, 3232-3239) (265 mg, inmol) 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 CH2CI2/MeOH (99.5:0.5) and the residue was crystallized from CH2Cl2fhexane to give 1- V3R)-2,3-dihydro-i1-methyl-2-oxo-5-phenyl-l1H-i1,4-benzodiazepin-3yl]-3-(2,4-dichlorophenyl)urea as a colorless solid, m.p. 215-216.5'C, [COD +76.20 (c=0.261, CHCl3).

dH (CDC13) 8.10 (1H, d, J 9.0 Hz), 7.65-6.95 (13H, mn), 5.50 (1H, d, J Hz), and 3.50 (3H, s).

Anal. Calcd. for C23H1I8C12N402.0.3H20: C, 60.22; H, 4.09; N, 12.21.

WO 98/00405 WO 9800405PCTIUS97/11131 Found: C, 60.28; H, 3.89; N, 12.10%.

EXAMPLE 59

CH

3 N 1..

0 (-)-3-Cyclohexyl-N- [(3R)-2,3-dihydro-l1-methyl-2-oxo-4-oxido-5p2henvi-l1H-i .4-benzodiazepin-3 -yllpropanamide 3-Chloroperoxybenzoic acid 0.32 g, 1.5 minol) was added to a solution of (+)-3-cyclohexyl-N-[(3R)-2,3-dihydro- 1-methyl- 1H-i ,4-benzodiazepin-3 -yllpropanamnide (0.60 g, mmol) in dichloromethane (25 mL) and the mixture was stirred at room temperature for 18 h. Further 3-chloroperoxybenzoic acid 0.1 g, nirol) 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 (MgSO4) and the solvent was evaporated under reduced pressure. The residue was recrystallized from toluene/hexane (65:3 5) to give (-)-3-cyclohexyl-N- [(3R)-2,3-dihydro-l1-methyl-2-oxo-4-oxido-5-phenyl-l1H- 1,4benzodiazepin-3 -yl] propanamide as colorless prisms, m.p. 222-224'C, dH (CDCI3) 7.7 1-7.23 (1lOH, in), 6.01 (1LH, d, J 9.3 Hz), 3.54 (3H, s), 2.48 (2H, in), and 1.76-0.89 (13H, in).

Anal. Calcd. for C25H29N303 5H20: C, 70.06; H, 7.06; N, 9.8 1.

Found: C, 70.10; H, 6.80; N, 9.79%.

WO 98/00405 PCT/US97/11131 82- EXAMPLE N-[2,3-Dihydro-1 -(2-dimethylaminoethyl)-2-oxo-5-phenyl- 1 H-1,4benzodiazepin-3-yll-3-(2.4-dichlorophenyl)propanamide Step A:

CH

3 N/CH3

N

-N

2,3-Dihydro- -(2-dimethylaminoethyl)-5-phenyl- H- ,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 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, 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 (MgSO4) and the solvent was evaporated under reduced pressure to give 2,3-dihydro-l-(2-dimethylaminoethyl)-5-phenyl-lH-l,4benzodiazepin-2-one (1.21 g, 93%).

WO 98/00405 PCT/US97/11131 83dH (CDC13) 7.63-7.16 (9H, 4.77 (1H, d, J 10.6 Hz), 4.41 (1H, m), 3.80 (1H, 3.78 (1H, d, J 10.6 Hz), 2.49 (2H, and 2.13 (6H, s).

Step B: CH31\ CH 3

N

IN NOH

NN

2,3-Dihydro- -(2-dimethylaminoethyl)-3-hydroxyimino-5-phenyl- H- 1,4-benzodiazepin-2-one 2,3-Dihydro-1 -(2-dimethylaminoethyl)-5-phenyl-1 H-1,4benzodiazepin-2-one (1.21 g, 3.9 mmol) was dissolved in toluene mL). The mixture was cooled to -78 °C and potassium t-butoxide (1.OM solution in t-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 0

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 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-1- (2-dimethylaminoethyl)-3-hydroxyimino-5-phenyl-1 H-1,4benzodiazepin-2-one (0.876 g, 66%) as a solid, m.p. 232-234°C.

dH (d6-DMSO) 10.90 (1H, 7.72-7.25 (9H, 4.40 (1H, 3.80 (1H, 2.50 (2H, and 1.85 (6H, s).

WO 98/00405 PCT/US97/11131 -84- Step C:

CH

3

NCH

3

N-

I NH 2

-N

3-Amino-2,3-dihydro-1 -(2-dimethylaminoethyl)-5-phenyl- 1 H-1,4benzodiazepin-2-one Ethyl isocyanate (320 mL, 287 mg, 4.0 nunol) was added to a mixture of 2,3-dihydro-l-(2-dimethylaminoethyl)-3-hydroxyimino-5phenyl-lH-1,4-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 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 (MgSO4) and evaporated under reduced pressure. The residue was dissolved in ethanol (100 mL), palladium on carbon 100 mg) was added and the mixture was shaken under hydrogen (50 for 4.5 h. Further palladium on carbon 100 mg) was added and the mixture was shaken under hydrogen (50 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 CH2C12/MeOH to give 3-amino-2,3-dihydro-1-(2-dimethylaminoethyl)- 5-phenyl-1H- ,4-benzodiazepin-2-one (180 mg, 17%).

dH (CDCl3) 7.75-7.17 (9H, 4.45 (1H, 4.40 (IH, 3.82 (1H, 2.47 (4H, and 2.08 (6H, s).

WO 98/00405 WO 9800405PCTIUS97/1 1131 85 Step E:

CH

3 NCH N-0 0 -N N C I C1 N- [2,3-Dihydro- 1 -(2-dimethylaminoethyl)-2-oxo-5 -phenyl- 1 H- 1,4benzodiazepin-3 -yll -3 -dichlorop~henvl)propanamide Triethylamine was added to a mixture of 3-amino-2,3dihydro-l1-(2-diniethylaminoethyl)-5-phenyl- 1H- 1,4-benzodiazepin-2one (180 mg, 0.6 mmol), 3-(2,4-dichlorophenyl)propanoic acid (131 mg, 0.6 mmol), 1 -dimethylaminopropyl)-3-ethylearbodiimide hydrochloride (115 mg, 0.6 mmol) and I1-hydroxyberzotriazole (81 mg, 0.6 mniol) 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 (MgSO4) and evaporated under reduced pressure. The residue was triturated with acetone and recrystallized from i- PrOH/MeOH to give N- [2,3 -dihydro-1- (2-dimnethylaminoethyl)-2-oxo-5phenyl- 1H-i ,4-benzodiazepin-3 -yl] ,4-dichlorophenyl)-propanamide as a solid, m.p. 199-201'C.

dH (CDCl3) 7.60-7.15 (13H, in), 5.50 (1H, d, J 8.0 Hz), 4.40 (1H, in), 3.80 (1H, in), 3.10 (211, t, J 7.5 Hz), 2.70 (2H, t, J 7.5 Hz), 2.40 (2H, in), and 2.05 (6H, Anal. Calcd. for C28H28C12N402: C, 64.25; H, 5.39; N, 10.70.

Found: C, 64.23; H, 5.40; N, 10.61 1%.

WO 98/00405 PCTIUS97/11131 -86- EXAMPLE 61

CH

3

O

\Nj H

CI

O *HCI [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-1,3-dihydro-l-methyl-5-phenyl- 2H-1,4-benzodiazepin-2-one Org. Chem. 1987, 52, 3232-3239) (265 mg, 1 mmol), E-1-chloro-4-(3-chloro-1-propenyl)benzene (281 mg, mmnol), potassium carbonate (276 mg, 2 mmol) and potassium iodide 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 (Na2SO4) 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 HCI (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 {N,N-bis[l-(4-chlorophenyl)propen-3-yl] amino }-1,3-dihydro- 1-methyl-5-phenyl-2H- 1,4benzodiazepin-2-one hydrochloride (235 mg, 39%) as a tan solid, m.p.

138-145 0 C, [ox]D (c=0.500, MeOH).

WO 98/00405 PCT/US97/11131 -87dH (d6-DMSO) 11.2 (1H, br 7.77-7.31 (17H, 6.85 (2H, br m), 6.54 (2H, 5.20 (1H, br 4.60-4.00 (4H, and 3.46 (3H, s).

Anal. Calcd. for C34H29C12N30.HCl.0. 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 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 N-[3-(4-chlorophenyl)propen-3yl] amino)-1,3-dihydro- 1-methyl-5-phenyl-2H- 1,4-benzodiazepin-2-one hydro-chloride (56 mg, 12%) as a yellow solid, m.p. 156-162 0 C, [a]D +350 (c=0.100, MeOH).

dH (d6-DMSO) 10.3 (1H, br 10.0 (1H, br 7.79-7.34 (13H, m), 6.78 (1H, d, J 15.9 Hz), 6.40 (1H, dt, Jd 15.9, Jt 9.0 Hz), 5.13 (1H, s), 4.00 (2H, and 3.46 (3H, s).

Anal. Calcd. for C25H22CIN30.HC1.0.10EtOH.0.40H20: 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 1-(2-bromoethoxy)-4-nitrobenzene or 4-chlorobenzenepropanol methanesulfonate for the E-l-chloro-4-(3-chloro-l-propenyl)benzene, the following compounds were prepared: WO 98/60405 WO 9800405PCTIUS97/11131 88 EXAMPLE 62

OH

3 0O\&N0 2 r-1

HCI

N0 2 I N,N-Bis [2-(4-nitrophenoxy)ethyl] amino) -1,3 -dihydro- 1 -methyl-5-phenyl-2H- 1,4-benzodiazepin-2-one hydrochloride m.12. 126-145'C. [ciD +5.00(0.100. CHCI3I).

dH (d6-DMSO) 8.20 (4H, d, J 9.2 Hz), 7.75-7.36 (9H, in), 7.08 (4H, d, J 9.2 Hz), 4.90 (1H, hr 4.50 (4H, hr 4.30-3.60 (5H, hr in), and 3.34 (3H, s).

Anal. Calcd. for C32H29N507 .HC1.O.1I5EtOH: C, 60.7 1; H, 4.87; N, 10.96.

Found: C, 60.70; H, 4.87; N, 10.70%.

EXAMPLE 63

HCI

WO 98/00405 WO 9800405PCTIUS97/11131 -89- I N- [3 -(4-Nitrophenoxy)ethyl] amino) -1 ,3 -dihydro- 1 -methyl 5-p2henyl-2H- 1 A-benzodiazepin-2-one hydrochloride m.p. 154-160'C, MaD +84.60(0.500, MeOH).

dH (d6-DMSO) 10.2 (1H, br 8.25 d, J 9.0 Hz), 7.83-7.4 1 (9H, in), 7.09 (2H, d, J 9.0 Hz), 5.21 (1 H, 4.57 (2H, mn), 3.70 (2H, in), 3.47 (3H, and 3.40 (1 H, in).

Anal. Calcd. for C24H22N404.HC1.0. 15EtOH.O.20H20: C, 61.13; H, 5.13; N, 11.74.

round: C, 61.12;, H, 4.92; N11.64%~.

EXAMPLE 64

OH

3 0 H Ii H HCI c {N-[3-(4-Chlorophenyl)prop- l -yl] amino) -1,3 -dihydro- 1 phenvl-2H- 1.4-benzodiazepin-2-one hydrochloride m.p. 167-168 0 C, MXD +20.80 (c=0.500, MeOH).

dH (d6-DMSO) 9.9 (2H1, br in), 7.78-7.26 (13H, in), 5.08 (1H, 3.45 (3H, 3.20 (1 H, in), 3.00 (1 H, in), 2.70 (2H, t, J 7.4 Hz), and 2.05 (2H, in).

Anal. Calcd. for C25H24C1N30.HC1: C, 66.08; H, 5.55; N, 9.25.

Found: C, 65.81; H, 5.49; N, 9.30%.

WO 98/00405 PCTIUS97/11131 EXAMPLE

OH

3

S

I0 N NH (+)-Phenylmethyl N- ,3-dihydro-l1-methyl-S -phenyl-2-thioxo- 1 H-i A-benzodiazepin-3-vllcarbamate A mixture of (+)-phenylmethyl N-[(3R)-2,3-dihydro- 1methyl-5-phenyl-2-oxo- IH-i ,4-benzodiazepin-3 -yl] carbamate (4.0 g, mmol) and 2,4-bis(4-methoxyphenyl)- 1,3 -dithia-2,4-diphosphetane-2,4disulfide (4.5 g, 11I mmol) in toluene (100 mL) was heated under reflux for 75 min. The mixture was cooled and the volume was reduced to 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 -dihydro- 1 -methyl-5-phenyl-2-thioxo- IH-i ,4-benzodiazepin-3-yl] carbamate as a solid, m.p. 129-13 1 C, [CC]D +22.50 (c=0.656, CHCl3).

dH (CDCl3) 7.65-7.26 (15H, in), 5.50 (lH, d, J 8.8 Hz), 5.14 (211, s), and 3.86 (311, s).

Anal. Calcd. for C24H21N302S.0.25H20: 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-12,3-dihydro-5-phenyl-2oxo- 1H-i ,4-benzodiazepin-3 -yl] carbamate for the (+)-phenytmethyl N- [(3R)-2,3-dihydro- 1-methyl-S -phenyl-2-oxo- 1H-i ,4-benzodiazepin- 3-yl]carbamate, the following compound was prepared: WO 98/00405 WO 9800405PCT/US97/11 131 91 EXAMPLE 66

HS

N 0 N H 0 Phenylmethyl N-[2,3 -dihydro-5 -phenyl-2-thioxo- 1H-i ,4-benzodiazepin- 3-yllcarbamate dH (d6-DMSO) 10.85 (1H, 8.42 (iR, d, J 8.6 Hz), 7.65-7.10 (14H, in), 5.10 (2H, and 5.05 (1WH d, J 8.6 Hz).

EXAMPLE 67

CH

3 S I S 0

N-

NHI NH

=N

3 -Cyclohexyl-N-(2,3-dihydro-l1-methyl-5-phenyl-2-thioxo- 1H- 1,4benzodiazepin-3-yl)p2ropanamide Hydrogen bromide was bubbled at room temperature through a solution of (+)-phenylmethyl N- -dihydro-l1-methyl 5-phenyl-2-thioxo- 1H-i ,4-benzodiazepin-3-yl]carbamate (0.9 g, 2.1 inmol), acetic acid (5 mL) and dichioromethane (5 mnL). After 2 hi., the solvent was evaporated under reduced pressure, ether was added and the WO 98/00405 PCT/US97/11131 -92solid 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 (Na2SO4) 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) and the residue was recrystallized from EtOAc/Hexane to give 3-cyclohexyl-N-(2,3dihydro-1 -methyl-5-phenyl-2-thioxo- 1 H- 1,4-benzodiazepin-3yl)propanamide as a solid, m.p. 219-221°C.

dH (CDC13) 7.95 (1H, br d, J 8.6 Hz), 7.65-7.30 (9H, 5.72 (1H, d, J 8.6 Hz), 3.87 (3H, 2.41 (2H, t, J 7.6 Hz), and 1.80-0.85 (13H, m).

Anal. Calcd. for C25H29N30S.0.25H20: 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- 1,4-benzodiazepin-3-yl]carbamate for the (+)-phenylmethyl 2,3-dihydro-l-methyl-5-phenyl-2-thioxo-lH-1,4-benzodiazepin-3yl]carbamate and an appropriate acid for the cyclohexanepropionic acid, the following compounds were prepared: WO 98/00405 WO 9800405PCTfUS97/11131 93 EXAMPLE 68 H S 0

N

NH

=N

3-Cyclohexyl-N-(2,3 -dihydro-5-phenyl-2-thioxo-l1H-i ,4-benzodiazepin- 3-yl~propanamide m.p. 113-119TC.

dH (CDCl3) 9.8 (LH, br 7.75-7.25 (10H, in), 5.75 (1H, d, J 8.1 Hz), 2.41 (2H, in), and 1.80-0.85 (13H, in).

Anal. Calcd. for C24H27N30S 8CH2C12: C, 62.91; H, 6.09; N, 8.87.

Found: C, 62.88; H, 5.70; N, 9.12%.

EXAMPLE 69 NH2 0-N 3 -Cyclohexyl-N-(2,3 -dihydro-2-hydrazono-5 -phenyl-l1H- 1,4benzodiazepin-3ylbpropanamide Hydrazine (53 mL, 56 mg, 1.8 inmol) was added to a solution of 3-cyclohexyl-N-(2,3 -dihydro- 1-methyl-S -phenyl-2-thioxo- WO 98/00405 PCTIUS97/11131 -94- 1H-1,4-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 (Na2SO4) 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 3cyclohexyl-N-(2,3-dihydro-2-hydrazono-5-phenyl- H-1,4-benzodiazepin-3yl)propanamide as a foam.

dH (CDCl3) 7.55-7.00 (11H, 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 C24H29N50.0.8CH30H.O. 15CH2C12: C, 67.82; H, 7.41; N, 15.85.

Found: C, 67.79; H, 7.46; N, 16.05%.

EXAMPLE

H

NOH

N-0 I N and (Z)-3-Cyclohexyl-N-(2,3-dihydro-2-hydroxyimino-5-phenyl- 1H-1, .4-benzodiazepin-3-yl)propanamide A mixture of 3-cyclohexyl-N-(2,3-dihydro-1-methyl-5 phenyl-2-thioxo-1H-1,4-benzodiazepin-3-yI)propanamide (740 mg, 1.83 mmol), hydroxylamine 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 WO 98/00405 WO 9800405PCTIUS97/11131 chromatography on silica gel, eluting with CH2Cl2IMeOH The residue recrystallized from ethyl acetate. The first isomer to crystallize was recrystallized from ethyl acetate to give(E) -3 -cyclohexy l-N-(2,3 dihydro-2-hydroxyimino-5-phenyl-l1H-i ,4-benzodiazepin-3 yl)propanamide as a solid, m.p. 196'C.

dH (d6-DMSO) 12.20 (1 H, 9.00 (1H1, d, J 8.0 Hz), 7.70-7.30 in), 5.45 (lH, d, J 8.0 Hz), 2.30 (2H, in), and 1.80-0.75 (13H, in).

The second isomer to crystallize was recrystallized from methanol to give -cyciohexyi-N-(2-,-dih*'ydro-2L-hyadroxyimino-5'phenyl-1H-1,4-benzodiazepin-3-yl)propanamide as a solid, in.p. 219'C.

dH (d6-DMSO) 9.95 (lH, 8.95 (l11, 8.75 (1H, d, J 8.0 Hz), 7.50- 7.00 (9H, in), 5.70 (1H, d, J 8.0 Hz), 2.25 (2H1, in), and 1.75-0.75 (13H, in).

Anal. Calcd. for C24H28N402: C, 71.26; H, 6.98; N, 13.85.

Found: C, 70.89; H, 6.99; N, 13.55%.

EXAMPLE 71 N 0

NH-I-

N

3-Cyclohexyl-N-(2,3-dihydro- 1 -methyl-S -phenyl- 1 H-i ,4-benzodiazepin- 3vD) propanamide Freshly prepared Raney nickel (400 mg) was added to a solution of 3-cyclohexyl-N- (2,3 -dihydro- 1-methyl-S -phenyl-2-thioxo- 1H-l ,4-benzodiazepin-3-yl)propanamide (200 mg, 0.5 minol) in ethanol mL) and the mixture was stirred at room temperature for 2 h. The WO 98/00405 PCTfUS97/11131 -96mixture was filtered and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2Cl2IMeOH (99.75:0.25) to give 3-cyclo.hexyl-N-(2,3 -dihydro-l1-methyl-5-phenyl- 11-1 ,4-benzodiazepin-3y1) propanamide as a foam.

dH (CDCI3) 7.60-6.80 (9H, in), 6.37 (1H, br d, J 6.6 Hz), 5.53 (lH, mn), 3.60 (2H, mn), 2.77 (3H, 2.21 (2H, t, J 8.0 Hz), and 1.85-0.80 (13H, in).

Anal. Caicd. for C25H31IN30.O.2C1-H2C12: 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-methyl-2-oxo-5-phenyl- 1H-thieno-[2,3-e]- 1.4diazepin-3 -yl'-3-(3-methyl-p2henyl)urea Step A: S NH 2 (2-Amino-3-thienvl)phenvlinethanone Triethylamnine (6.8 inL, 4.94 g, 49 nimol).was added to a heated (33'C) mixture of b-oxobenzenepropanenitrile (18.6 g, 12R nimol) and 1,2-dithiane-2,5-diol (9.8 g, 64 inmol) in ethanol (120 niL) and the mixture was stirred at 5OC' 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 (0.5 aqueous sodium hydroxide (1IM) and brine, WO 98/00405 PCT/US97/11131 97 dried (Na2SO4) and the solvent was evaporated under reduced pressure.

The residue was recrystallized from acetonitrile (150 mL) to give (2amino-3-thienyl)-phenylmethanone as an orange solid (5.7 g, 44%).

dH (CDC13) 7.70-7.35 (5H, 6.95 (2H, br 6.90 (1H, d, J 6.3 Hz), and 6.15 (1H, d, J 6.3 Hz).

Step B: H O0 S N-

-N

2.3-Dihydro-5-phenvl- 1H-thieno[2.3-el-l .4-diazepin-2-one A solution of 1,3-dihydro-1,3-dioxo-2H-isoindole-2-acetyl chloride (8.6 g, 38 mmol) in dichloromethane (20 mL) was added slowly to a cooled mixture of (2-amino-3-thienyl)phenylmethanone (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 saturated aqueous sodium hydrogen carbonate and brine. The mixture was dried (Na2SO4) 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,3dihydro-l,3-dioxo-2H-isoindole-2-acetamide as a solid (9.8 g, 76%).

A mixture of N-(3-benzoylthien-2-yl)-1,3-dihydro-1,3dioxo-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 I WO 98/00405 PCT/US97/11131 98 combined organic fractions were washed with brine, dried (Na2SO4) and the solvent was evaporated under reduced pressure. Acetic acid (300 mL) was added and the mixture was heated under reflux for 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 (Na2SO4) and the solvent was evaporated under reduced pressure to give 2,3-dihydro-5phenyl-iH-thieno[2,3-e]-,4-diazepin-2-one as a foam (3.5 g, 52%).

dH (CDC13) 9.75 (1H, br 7.90-7.30 (5H, 6.87 (1H, d, J 6.0 Hz), 6.82 (1H, d, J 6.0 Hz), and 4.45 (2H, s).

Step C: S N-

-N

2.3-Dihydro-l-methvl-5-phenvl-lH-thieno[2.3-el-1.4-diazepin-2-one Sodium hydride (60% dispersion in mineral oil, 757 mg, 11.3 mmol) was added to a cooled solution of 2,3-dihydro-5phenyl- H-thieno[2,3-e]-1,4-diazepin-2-one (2.61 g, 10.8 mmol) in DMF (7 mL). Further DMF (10 mL) 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 (Na2SO4) and evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with WO 98/00405 PCT/US97/11131 -99- CH2Cl2/MeOH (95:5) to give 2,3-dihydro- 1-methyl-5-phenyl- 1Hthieno[2,3-e]-1,4-diazepin-2-one (1.5 g, 54%).

dH (CDC13) 7.67-7.35 (5H, 7.00 (1H, d, J 6.0 Hz), 6.85 (1H, d, J Hz), 4.45 (2H, br and 3.50 (3H, s).

Step D: 0 S-NH2

-N

3-Amino-2,3-dihydro- -methyl-5-phenyl-1H-thieno[2,3-e]-1,4-diazepin- 2-one 2,3-Dihydro-l-methyl-5-phenyl-1H-thieno[2,3-e]-1,4diazepin-2-one (1.5 g, 5.8 mmol) was dissolved in toluene (30 mL).

The mixture was cooled to -10 0 C and potassium t-butoxide (1.7 g, 15.1 mmol) was added. The mixture was stirred at -10 0 C for 15 min., then isoamyl nitrite (1.0 mL, 0.87 g, 7.4 mmol) was added. The mixture was stirred at -10 0 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 (Na2SO4) 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- H-thieno[2,3-e]-1,4diazepin-2-one (0.80 g, 48%).

2,3-Dihydro-1 -methyl-3-hydroxyimino-5-phenyl-1 H-thieno [2,3-e]-1,4-diazepin-2-one (0.80 g, 2.8 mmol) was dissolved in ethanol mL) and Raney nickel (2 g) was added. The mixture was shaken under hydrogen (50 for 5 days, adding further Raney nickel g) in portions. The mixture was filtered and the solvent was evaporated WO 98/00405 PCT/US97/11131 -100under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with CH2CI2INeOH to give 3amino-2,3 -dihydro-l1-methyl-5-phenyl- 1H-thieno[2,3 -el-1 ,4-diazepin-2one (248 mg, 33%).

dH (CDCI3) 7.50-7.30 (5H, in), 7.05 (1lH, d, J 6.0 Hz), 6.85 (1lH, d, J Hz), 4.57 (lH1, 3.55 (3H, and 1.70 (2H, br s).

Step E:

OH

3 S NO

H

3 1 -Dihydro- 1-methyl-2-oxo-5-phenyl- 1 H-thieno -1,4-diazepin- 3 3 -(3-methylphenyl~urea 3-Methyiphenylisocyanate (60 mL, 62 mg, 0.46 mmol) was added to a solution of 3-amino-2,3-dihydro-l-methyl-5-phenyl-lHthieno[2,3-e]-1,4-diazepin-2-one (124 mg, 0.46 minol) in tetrahydrofuran (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 1 -(2,3-dihydro- 1-methyl- 1H-thieno [2,3 1,4-diazepin-3 -yl)-3 -methylphenyl)urea as a solid (94 mg, m.p. 128-130TC.

dH (CDCl3) 8.70 (lH, 7.65-6.75 (12H, in), 5.55 (1H, d, J 9.0 Hz), 3.55 (3H, and 2.30 (3H, s).

Anal. Calcd. for C22H20N402S.0.25H20: C, 64.62; H, 4.99; N, 13.70.

Found: C, 64.68; H, 4.96; N, 13.70%.

WO 98/00405 PCTIUS97/11131 -101 EXAMPLE 73

OH

3

I

3-Cyclohexyl-N-(2,3-dihydro-l1-methyl-2-oxo-5-phenyl- 1H-thieno[2,3 e] -1.4-diazepin-3 -yl)propanamide Triethylamine (75 mL, 54 mg, 0.54 mmol) was added to a mixture of 3-amino-2,3-dihydro-l1-methy1-5-phenyI~1H-thieno[2,3 1 ,4-diazepin-2-one (82 mg, 0.3 mmol), cyclohexanepropanoic acid (52 mL, 47 mg, 0.3 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide 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 saturated aqueous sodium hydrogen carbonate and brine, dried (Na2SO4) and the solvent was evaporated under reduced pressure. The residue was purified by flash colunm chromatography on silica gel, eluting with EtOAc/Hexane to give 3 -cyclohexyl-N-(2,3 -dihydro-l1-methyl-2-oxo-5-phenyl- 1Hthieno[2,3-e]-1 ,4-diazepmn-3-yl)propanamide as a solid (56 mg, 46%).

M.P. 189-190 0

C.

dH (CDCI3) 7.65-6.85 (8H, in), 5.65 (1 H, d, J 8.0 Hz), 3.55 (3H, s), 2.40 (2H, t, J 7.0 Hz), and 1.80-0.85 (13H, mn).

Anal. Calcd. for C23H27N302S.0.5H20: C, 66.00; H, 6.74; N, 10.04.

Found: C, 66.25; H, 6.76; N, 9.83%.

It.

WO 98/00405 PCT/US97/11131 -102 EXAMPLE 74 H 0 N 0

NH--

N

3-Cyclohexyl-N-(5-cyclohexyl-2,3-dihydro-2-oxo- H-1,4benzodiazepin-3-vyl) propanamide Phenylmethyl N-[5-cyclohexyl-2,3-dihydro-2-oxo-1H-1,4benzodiazepin-3-yl]carbamate (150 mg, 0.38 mmol) was dissolved in hydrogen bromide in acetic acid 0.5 mL). After 2 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 stirred 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 saturated aqueous sodium hydrogen carbonate, water and brine, dried (Na2SO4) and the solvent was evaporated under reduced pressure. The residue was recrystallized from i-PrOH to give 3cyclohexyl-N-(5-cyclohexyl-2,3-dihydro-2-oxo-1H-1,4-benzodiazepin- 3-yl)propanamide as a solid, m.p. 133-138 0

C.

dH (CDC13) 7.85 (1H, br 7.62-6.95 (5H, 5.40 (1H, d, J 8.7 Hz), 2.77 (1H, 2.34 (2H, and 2.05-0.75 (23H, m).

Anal. Calcd. for C24H33N302.0.7C3H70H: C, 71.64; H, 8.89; N, 9.60.

WO 98/00405 PCT/US97/11131 103- Found: C, 71.28; H, 8.70; N, 9.82%.

EXAMPLE

CH

3 o Cl HNN i C1 H

CI

(+)-N-[(3R)-7-Amino-2,3-dihydro- -methyl-2-oxo-5-phenyl- 1H-1,4benzodiazepin-3-yll-3-(2.4-dichlorophenyl)propanamide Step A: To a mixture of 3(R)-amino- phenyl-2H-1,4-benzodiazepin-2-one 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 (2.1.2 g, 21.0 mmol) in concentrated sulfuric acid (6 mL). The mixture was stirred with cooling for 2 then stirred at ambient temperature for 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 (Na2SO4) 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 n-butyl chloride (30 mL) and the solvent was evaporated under reduced pressure to give an inseparable mixture of 3(R)-amino-1,3-dihydro-l-methyl-7-nitro-5phenyl-2H-1,4-benzodiazepin-2-one and 3(R)-amino-l,3-dihydro-1- WO 98/00405 PCT/US97/11131 -104methyl-7-nitro-5-(2-nitrophenyl)-2H-1,4-benzodiazepin-2-one (3.81 g) in a 3:1 ratio as a yellow solid.

dH (CDC13) (mononitro compound) 8.43 (1H, dd, J 9, 3 Hz), 8.23 (lH, d, J 3 Hz), 7.59 (2H, 7.52 7.44 4.47 3.53 and 2.42 (2H, br (dinitro compound) 8.49 (1H, dd, J 9, 3), 8.42 (1H, 8.18 (lH, d, J 3 Hz), 8.01 (1H, 7.67 (1H, t, J 6 Hz), 7.6-7.4 (2H, 4.52 3.56 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-(2,4dichlorophenyl)propionyl chloride (520 mg, 100%). To a solution of mixed 3(R)-amino-1 ,3-dihydro-1 -methyl-7-nitro-5-phenyl-2H-1,4benzodiazepin-2-one and 3(R)-amino-1,3-dihydro-1-methyl-7-nitro-5- (2-nitrophenyl)-2H-1,4-benzodiazepin-2-one (621 mg, 2 mmol) and triethylamine (0.305 mL, 2.2 mmol) in methylene chloride 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- 1 -methyl-7-nitro- 2-oxo-5-phenyl-1H-1 ,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)propanamide and (+)-N-[(3R)-2,3-dihydro-1-methyl-7-nitro-2-oxo-5- (2-nitrophenyl)-1H-1 ,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)propanamide (850 mg, 84%) in a 3:1 ratio as a solid white foam.

dH (CDCl3) (mononitro compound) 8.45 (1H, dd, J 9, 3 Hz), 8.25 (1H, d J 3 Hz), 7.54 (3H, 7.45 (2H, 7.38 (1iH, d, J 2 Hz), 7.26-7.18 (4H, 5.50 (IH, d, J 8 Hz), 3.52 (3H, 3.10 (2H, and 2.70 (2H, (dinitro compound) 8.51 (1H, dd, J 9, 3 Hz), 8.40 (1H, 8.21 (1H, dJ 3 Hz), 7.98 (1H, 7.68 (IH, t, J 6 Hz), 7.60 (1H, 7.44 WO 98/00405 PCT/US97/11131 -105- (1H, 7.26-7.15 (4H, 5.52 (IH, d, J 8 Hz), 3.55 (3H, 3.10 (2H, and 2.70 (2H, m).

Step C: To a solution of mixed N-[(3R)-2,3-dihydro-1-methyl- 7-nitro-2-oxo-5-phenyl-1H-1 ,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)propanamide and (+)-N-[(3R)-2,3-dihydro- 1-methyl-7-nitro-2oxo-5-(2-nitrophenyl)- 1H- 1,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)propanamride (770 mg, 1.5 mmol) in acetic acid (6 mL) was added dropwise in portions over 1.5 h. a solution of 15% titanium (III) 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 diluted with water (80 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic fractions were washed with brine, dried (Na2SO4) and the solvent was evaporated under reduced pressure. The residue was purified by flash column chromatography on silica gel, eluting with ethyl acetate/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- 1-methyl-2-oxo- 5-phenyl-1 H-1,4-benzodiazepin-3-yl] -3-(2,4-dichlorophenyl)propanamide (413 mg, 57%) as a pale yellow solid, m.p. 179-180 0

C,

[o]D +60.20 0.500, CHCl3).

dH (CDCl3) 7.60 (2H, d, J 7 Hz), 7.49-7.36 (5H, m) 7.24 (1H, d, J 9 Hz), 7.17 (2H, 6.99 (IH, dd, J 9, 3 Hz), 6.64 (1H,d, J 3 Hz), 5.54 (1H, d, J 8 Hz), 4.80-3.50 (2H, br 3.39 (3H, 3.09 (2H, t, J 8 Hz), and 2.68 (2H, dt, Jd 3, Jt 8 Hz).

Anal. Calcd. for C25H22C2N402: 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- 1-methyl-2-oxo-5- (2-aminophenyl)- 1 H-1 ,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)propanamide (114 mg, 15%) as a pale yellow solid, m.p. 188-189 0

C,

WO 98/00405 PCT/US97/11131 -106 [O]D +50.0° (c=0.100, MeOH).

dH (CDC13) 7.36 (2H, 7.25 (1H, d, J 9 Hz), 7.15 (3H, 7.00 (1H, 6.88 (2H, 6.79 (1H, 6.60 (1H, bs), 5.52 (1H, d, J 8 Hz), 4.10-2.80 (4H br 3.40 (3H, 3.09 (2H, t, J 8 Hz), and 2.69 (2H, m).

Anal. Calcd. for C25H23Cl2N502.0.05EtOAc: C, 60.43; H, 4.71; N, 13.99.

Found: C, 60.79; H, 4.74; N, 13.83%.

EXAMPLE 76

CH

3 N O Cl

CH

3

SO

2

NHN

H C1 (+)-N-[(3R)-2,3-Dihydro-1 -methyl-2-oxo-5-phenyl-7-methanesulfonamido- 1H- 1,4-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-2oxo-5-phenyl-1H-1,4-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 (Na2SO4) 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 WO 98/00405 WO 9800405PCTIUS97/11131 107 dried in vacuo to give (+)-N-[(3R)-2,3-dihydro-1-methyl-2-oxo-5phenyl-7-methanesulfonamido- 1H-i ,4-benzodiazepin-3 -yl] -3 dichlorophenyl)propanamnide (152 mg, 68%) as a white solid, m.p. 130- 148 0 C, [M~D +111.60 (c=0.500, CHCl3).

dH (CDCI3) 7.55-7.32 (9H, in), 7.24 (2H, dd, J 10, 2 Hz), 7.17 (1H, dd, J 9, 2 Hz), 7.05 (1LH, d, J 3 Hz), 5.49 (1 H, d, J 8 Hz), 3.41 (3H, 3.08 (2H, t, J 8 Hz), 2.97 (3H, and 2.71 (2H1, dt, Jd 3, Jt 8 Hz).

Anal. Calcd. for C26H24C12N404S: C55.82; H, 4.32; N, 1.01.

Found: C, 56.12; H, 4.47; N, 9.89%.

EXAMPLE 77

CH

3 0 N- 0 C1 N( N N N I 1 -1: H c1

*HCI

N-(2,3 -Dihydro-l1-methyl-2-oxo-5-phenyl- 1H-pyrido[4,3-e] -1,4diazep~in- 3-yl)-3 -(2,4-dichlorophenylbpropanamide hydrochloride Step A: To a solution of 2,3-dihydro-1-methyl-5-phenyl-1Hpyrido[4,3-e]-1,4-diazepine-2-one (Q Med. Chem.. 1965, 8, 722-724) (1.63 g, 6.5 inmol) in toluene (32 mL) under argon cooled to (ice/methanol bath) was added potassium t-butoxide (1.83 g, 16.3 mmol). The resulting purple suspension was stirred 15 min. at -201 C and isoamyl nitrite (1.05 mL, 7.8 inmol) was added. The mixture was stirred at -20'C for 30 min., then poured into a mixture of water mL), acetic acid (3 mL), and ethyl acetate (65 mL). The mixture was WO 98/00405 PCT/US97/11131 -108 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 (Na2SO4), 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-3hydroxyimino-l-methyl-5-phenyl-lH-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 (1UH, bs 8.73 (114, d1 7 Hz7) 8R62 (1H. 7.80 (2H, dd, J 7, 1 Hz), 7.59 (1H, 7.48 (2H, 7.26 (1H, d, J 7 Hz), and 3.50 (3H,s).

Step B: A mixture of 2,3-dihydro-3-hydroxyimino- phenyl-lH-pyrido[4,3-e]-1,4-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 (Na2SO4) and the solvent was evaporated under reduced pressure to give 3-amino-2,3-dihydro- -methyl-5-phenyl-lH-pyrido[4,3-e]-1,4diazepine-2-one (276 mg, as a yellow solid, m.p. 109-123 0

C.

dH (CDC13) 8.72 (1H, d, J 6 Hz), 8.58 (1H, 7.61 (2H, 7.51 (1H, 7.43 (2H, 7.26 (1H, 4.47 (lH 3.50 (3H, and 2.1 (2H, bs).

High res. mass spectrum: Theoretical mass for C15H14N40 267.124586. Measured mass 267.123654.

WO 98/00405 PCT/US97/11131 -109- Step C: A solution of dicyclohexylcarbodiimide (87 mg, 0.42 mmol) in methylene chloride (0.17 mL) was added to a solution of 3amino-2,3-dihydro-l -methyl-5-phenyl-lH-pyrido[4,3-e]-1,4-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 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 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 (Na2SO4) and the solvent was evaporated under reduced pressure. The residue was suspended in ethanol (2 mL) and ethanolic HCI (6.8 M, 0.147 mL) was added. The mixture was stirred, the resulting precipitate was collected and dried in vacuo to give N-(2,3-dihydro-l-methyl-2-oxo-5-phenyl-1H-pyrido[4,3e]-1,4-diazepin-3-yl)-3-(2,4-dichlorophenyl)propanamide hydrochloride (32 mg, 18%) as a white solid, m.p. 218-219 0

C.

dH (d6-DMSO) 9.38 (1H, d, J 8 Hz), 8.86 (1H, bs), 8.59 (1H bs), 7.79 (1H, d, J 6 Hz), 7.56 (3H, 7.51 (2H, 7.39 (2H, 7.25 (1H, 7.16 (1H, 5.37 (1H, 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 C24H20C12N402.HC1: C, 57.22; H, 4.20; N, 11.12.

Found: C, 56.87; H, 4.18; N, 11.09%.

WO 98/00405 PCT/US97/11131 -110- EXAMPLE 78

CH

3 NN N -N N

H

N-(2,3-Dihydro-1 -methyl-2-oxo-5-phenyl-1 H-pyrido[4,3-e]-1,4diazepin-3-vl)-3-(cyclohexvl)propanamide A solution of dicyclohexylcarbodiimide (87 mg, 0.42 mmol) in methylene chloride (0.17 mL) was added to a solution of 3amino-2,3-dihydro-l-methyl-5-phenyl-1H-pyrido[4,3-e]-1,4-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 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 The purified material was stirred under chloroform mL) with potassium carbonate (0.1 g) and water (2 drops) for 5 min.

The mixture was dried (Na2SO4) and the solvent was evaporated under reduced pressure. The residue was crystallized from toluene to give N- (2,3-dihydro-l-methyl-2-oxo-5-phenyl-lH-pyrido[4,3-e]-1,4-diazepin-3yl)-3-(cyclohexyl)-propanamide (47 mg, 33%) as a white crystalline solid, m.p. 170-173°C.

dH (CDC13) 8.75 (1H, d, J 6 Hz), 8.61 (1H, 7.58 (2H, 7.52 (1H, 7.45 (2H, 7.31 (IH, d, J 6 Hz), 7.21 (1H, d, J 8 Hz), 5.54 (1H, d, J 8 Hz), 3.51 (3H, 2.39 (2H, 1.73 (4H, 1.63 (3H, 1.85- 1.12 (4H, and 0.94 (2H, m).

Anal. Calcd. for C24H28N402.0.10PhCH3: C, 71.70; H, 7.02; N, 13.54.

WO 98/00405 WO 9800405PCTIUS97/11131 III 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

CH

3 N- 0 N N H

CF

3 N-(2,3-Dihydro- 1-methyl-2-oxo-5-phenyl- 1H-pyridoll4,3-e]- 1,4diazep~in-3 -yl)-3 -(4-trifluoromethylphenyl)p2rop~anamide m.p. 191-192'C.

dH (CDCl3) 8.76 (1H, d, J 6 Hz), 8.61 (1H, 7.56 (4H1, in), 7.52 (1H, in), 7.42 (2H1, d, J 7 Hz), 7.38 (21-1, in), 7.30 (LH, d, J 6 Hz), 7.22 (1H, d, J 8 Hz), 5.51 (1H, d, J 8 Hz), 3.50 (3H, 3.09 (214, t, J 8 Hz), and 2.73 (2H, t, J 8 Hz).

Anal. Calcd. for C25H21F3N402.O.2OPhCH3: C, 65.39; H, 4.70; N, 11.56.

Found: C, 65.69; H, 4.64; N, 11.95%.

WO 98/00405 PCT/US97/11131 -112- EXAMPLE

CH

3 N N CI N

N

-N

H

CI

N-(2,3-Dihydro-l -methyl-2-oxo-5-phenyl-1 H-pyrido[3,4-e]-1,4diazepin-3-yl)-3-(2.4-dichlorophenvl)propanamide Step A: To a solution of 2,3-dihydro-1-methyl-5-phenyl-1Hpyrido[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 0

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 0 C for 30 min., then poured into a mixture of water 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 (Na2SO4), 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 2,3dihydro-3-hydroxyimino- 1-methyl-5-phenyl-lH-pyrido[3,4-e]-l,4diazepine-2-one (1.60 g, 100%) as a tan foam.

dH (CDC13) 8.77 (1H, 8.50 (IH, d, J 4 Hz), 7.81 (2H, dd, J 8, 1 Hz), 7.60 (1H, 7.49 (3H, 7.32 (1H, d, J 5 Hz), and 3.55 (3H,s).

WO 98/00405 PCT/US97/11131 113 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-1 -methyl-5-phenyl-1H-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 ashed with brine (30 mL), dried (Na2SO4), 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 mL) with potassium carbonate (0.3 g) and water (2 drops) for 5 min.

The mixture was dried (Na2SO4) 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-1-methyl-5phenyl-1H-pyrido[3,4-e]-1,4-diazepine-2-one (241 mg, 16%) as a yellow solid, m.p. 94-118°C.

dH (CDC13) 8.79 (1H, 8.48 (1H, d, J 5 Hz), 7.62 (2H, dd, J 8, 1 Hz), 7.51 (1H, 7.45 (2H, 7.24 (1H, dd, J 5, 1 Hz), 4.47 (1H 3.55 (3H, and 2.2 (2H, bs).

Anal. Calcd. for C15H14N40.0.25(C2H5)20: 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 C15H14N40 267.124586. Measured mass 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 WO 98/00405 PCT/US97/11131 -114evaporated under reduced pressure to give 3-(2,4-dichlorophenyl)propionyl chloride (52 mg, 100%). To a solution of 3-amino-2,3dihydro-l-methyl-5-phenyl-1H-pyrido[3,4-e]- ,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 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- 1H-pyrido[3,4-e]-1,4-diazepin-3-yl)-3-(2,4-dichlorophenyl)propanamide (38 mg, 38%) as a white crystalline solid, m.p. 220-221 0

C.

dH (CDCl3) 8.81 (1H, 8.52 (1H, d, J 5 Hz), 7.56 (2H, dd, J 7, 2 Hz), 7.51 (1H, 7.44 (2H, d, J 6 Hz), 7.40 (1H, 7.27 (2H, 7.18 (2H, dd, J 8, 2 Hz), 5.48 (1H J 8 Hz), 3.55 (3H, 3.10 (2H, t, J 7 Hz), and 2.71 (2H, dt, Jd 2 Jt 8 Hz).

Anal. Calcd. for C24H20C12N402.0.25PhCH3: 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-1-methyl-2-oxo-5-isopropyl-1H- ,4-benzodiazepin-3yll-3-(2.4-dichlorophenvl)propanamide Step A:

CH

3 CH3

\O

N (BOC) 2 0 N A NaH H THF

N\

0 n BOC WO 98/00405 PCT/US97/11131 -115- To a solution of the benzodiazepine (1.0 g, 5.3 mmol) in THF (20 mL) at -78 0 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 0 C for 1/2 hour. The reaction was then allowed to warm to 25 0

C

and stirred for 2 hours before quenching into cold aq. NH4C1 and extracting the product into ethyl acetate (3x50 mL). Concentration of the dried (Na2SO4) extracts gave an oil which was passed through silica (EtOAc/hexane) to give 1.35 g product !H NMR (CDC13) d: 1.60 9H), 3.40 3H), 3.95 (brd, 4.80 (brd, 1H), 7.20 1H), 7.30 1H), 7.60 1H), 7.92 1H).

Step B: CH3

CH

3

CH

3 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 NH4Cl mL), and extracted with ethyl acetate (2x200 mL). The organic extracts were concentrated and chromatographed on silica EtOAC/hexane) to give 1.55 g of product.

1 H NMR (CDC13) d: 1.14 3H), 1.19 3H), 1.40 9H), 3.13 (s, 3H), 3.2-3.8 3H), 5.45 (brs, 1H), 7.28 (dt, 1H), 7.48 (dt, 1H), 7.56 (dt, IH), 7.72 (dd, 1H).

WO 98/00405 PCT/US97/11131 -116- Step C:

CH

3

C

H

3 I BOC 1 0 N N 1) HCI, EtOAc NJ S0 H 2) LiOH, H20 O -N To a 0°C solution of the isopropylphenone (1.55 g) in ethyl acetate was added anhydrous HCI gas over 90 min. The reaction was then concentrated in vacuo to give a solid which was dissolved in 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 HCI and product extracted into ethyl acetate. The organic extracts were dried (Na2SO4), filtered and concentrated to give a solid 1.22 g, 100%.

1 H NMR (CDC13) d: 0.95 3H), 1.30 3H), 3.16 (septet, 1H), 3.36 3H), 3.60 1H), 4.60 1H), 7.2-7.3 2H), 7.45-7.55 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 (Na2SO4) extracts gave 2.60 gms Step F: WO 98/00405 WO 9800405PCTIUS97/11131 -117- The anine was coupled with 3-(2,4-dichlorophenyl)propionic acid as described in Example 43 to yield N-(2,3-dihydro-1I- 1H-i ,4-benzodiazepin-3-yl)-3-(2,4dichlorophenyl)propanamide.

1 H NMR (CDCI3) d: 0.92 3H1), 1.25 3H), 2.65 (dt, 2H), 3.05 (t, 2H), 3.15 (SepT, 1H1), 3.40 3H), 5.38 1H), 7.0-7.6 (in, 8H).

The following compounds were prepared in a similar mnanner as described in Example 801, using the appropriate Grignard reagent in place of isopropyl magnesium chloride.

.EXAMPLE 82 N- [2,3 -dihydro-l1-methyl-2-oxo-5-isopropyl- 1H-i ,4-benzodiazepin-3yl] -3 -cyclohexylprop~anamide m.p. 164-165'C CHIN: Anal. Calcd. for C221131IN302: C, 71.51; H, 8.46; N, 11.37 Observed: C, 71.72; H, 8.39; N, 11.32 EXAMPLE 83 N- [2,3-dihydro-lI-methyl-2-oxo-5-isopropyl- 1H-i ,4-benzodiazepin-3yl] -3 -(4-trifluoromethylphenyl)p2rop~anamide m.p. 187-188 0

C

lI i NMR (CDC13) d: 0.92 3H), 1.25 3H), 2.66 (dt, 2H), 3.04 (t, 2H), 3,15 (SepT, 1H), 3.40 3H), 5.38 1H), 7.14 (brd, 1H), 7.25- 7.6 (in, 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: WO 98/00405 PCTIUS97/11131 118-

CH

3

CH

3 0 I 0 N-

N

O N-OH ONH 2 -N -N O O 1 2 To a solution of the oxime 1 (1.28 g, 0.0048 mole) in (130 ml) and THF (65 ml) was added sodium dithionite (Na2S204) (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 Na2SO4, filtered, and concentrated to give an oil 1.0 The oil was chromatographed on silica using ethyl acetate followed by 10% methanol/methylene chloride to give pure amine 0.778g 1H NMR (DMSO) d 3.32 3H), 4.30 1H), 6.64 d, 1H), 6.76 1H), 7.35 (dt, 1H), 7.58-7.74 3H), 7.88 1H).

EXAMPLE 84 N-[2,3-dihydro-1 -methyl-2-oxo-5-(2-furanyl)-1 H- 1,4-benzodiazepin-3yll-3-cvclohexylpropanamide m.p. 168-169 0

C

CHN: Anal. Calcd. for C23H27N303: C, 70.21; H, 6.92; N, 10.68 Observed: C, 70.15; H, 6.67; N, 10.64 WO 98/00405 PCT/US97/11131 -119- EXAMPLE N-[2,3-dihydro-lI-methyl-2-oxo-5-(2-furanyl)- 1H-i ,4-benxodiazepin-3vii -3 -(4-trifluoromethylphenyl)p2ropanarnide m.p. 155-157'C CHN: Anal. Calcd. for C24H20N303F3: C, 63.29; H, 4.432; N, 9.23 Observed: C, 63.22; H1, 4.44; N, 9.07 EXAMPLE 86 N-[2,3-dihydro- 1 -methyl-2-oxo-5-(2-furanyl)- luH-i ,4-benzodiazepin-3yll -3 -(2,4-dichlorohenvl)p2rop~anamide m.p. 132-133'C CHN: Anal. Calcd. for C23H1I9N303C12 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-l1-methyl-2-oxo-5-(3-furanyl)- lu-I,4-benzodiazepin-3yll -3 -cyclohexylp~rop~anamide m.p. 199-200'C III NMR (CDCl3) d: 0.9-1.8 (brm, 3H), 2.38 2H), 3.42 3H), 5.55 (brd, 1H), 6.90 1H), 7.2-7.77 (in, 7H) EXAMPLE 88 N-[2,3-Dihydro-l1-methyl-2-oxo-5-(3 -furanyl)- 1H-i ,4-benzodiazepin-3yll -3 -(4-trifluoromethylphenyl)propan amide m.p. 213-214'C I H NMR (CDCl3) d: 2.71 (dt, 2H), 3.05 2H), 3.42 3H1), 5.72 (d, I 6.82 (brS, I 7.2-7.7 (in, 1 I H) WO 98/00405 WO 9800405PCT/US97/1 1131 -120- EXAMPLE 99 N-[2,3-Dihydro- 1 -methyl-2-oxo-5-[2'-(4,4-dimethyl-2-oxazolinyl)phenyl- -1 H- 1,4-benzodiazep~in-3-vll -3-(22A-dichlorophenyL)prop anamide The subject compound was prepared substantially as described in Example 81.

m.p. 19Y.-1I95'C CHN: Anal. Calcd. for C30H2SN403C12 C, 63.95; H, 5.01; N, 9.94 Found: C, 63.70; H, 5.01; N, 9.96 EXAMPLE N-[2,3 ,4,5-Tetrahydro-l1-methyl-2-oxo-5-isopropyl- 1H-I ,4-benzodiazepin-3v1] -3 -cvclohexylp~rop~anamide

CH

3 H, 10% Pd/C 0N MeOH N

H

O H 3 OH 3

CH

3 N- 0

H

0

N

NHH

CHH

OH CH 3 A solution of N-[2,3-dihydro- I -methyl-2-oxo-5 -isopropyl- WO 98/00405 WO 9800405PCT/US97/1 1131 -121 1 H-i ,4-benzodiazepin-3 -yl] -3 -cyclohexyipropanamide (50 mg) in methanol (10 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-l1-methyl-2-oxo-5 -isopropyl- 1H-I ,4-benzodiazepin- 3-yl]-3-cyclohexylpropanamide.

CHN: Anal. Calcd. for C22H33N302 C, 71.12; H, 8.95; N, 11.31 O b se rv ed: C, 7 0. 98; 18.97,IN, JI1.i11 m.p. I1I4-115'C EXAMPLE 91 N- [2,3-dihydro-l1-methyl-2-oxo-5 -methyl-i H-i ,4-benzodiazepin-3 -yl] 3-(2,4-dichlorop~henyl)propanamide Step A:

CH

3 00H 3 N-OH

H

H 00

H

N OH 3

OCH

3 C) H B Q NBZ N H-N H

OH

3

OH

3 To CBZ-benzodiazepine (250 mg, 0.776 mmol) in toluene 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 (CDCl3) d: 2.50 3H), 3.42 3H), 5.12-5.20 (in, 3H), 6.62 1H), 7.25-6.4 (mn, 7H1), 7.5-7.6 (in, 211).

WO 98/00405 PCT/US97/11131 122- Step B: N N

O

0 N N.CBZ O3 NH 2 -N H -N

CH

3 CH 3 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 CH2C12 mL) and organic layer separated, dried (Na2SO4), filtered and concentrated to an oil (172 mg, 110%).

1 H NMR (CDC13) d: 2.42 3H), 3.05 (brs, 2H), 3.40 3H), 4.40 (s, 1H), 7.2-7.6 4H).

Step C: HO i NH C l Ob NH 2 QH N EDC, HOBT, TEA, N DMF

CH

3

CH

3 0 NH

CI

-N

CH

3 WO 98/00405 WO 9800405PCTIUS97/11131 123 N- [2,3-dihydro- 1 -methyl-2-oxo-5 -methyl- I Hl-I ,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 CHIN: Anal. Calcd. for C20H19N302C12 C, 59.42; H, 4.74; N, 10.39 Observed: C, 59.50; H, 4.74; N, 10.44 1 H NMR (CDCI3) d: 2.49 (brs, 3H), 2.65 (dt, 2H), 3.05 2H), 3.42 (s, EXAMPLE 92 N-[2,3-Dihydro-l1-methyl-2-oxo-[4,5 -oxo- 1,3 -dihydro-2Hisoindole)- 1 H-I ,4-benzodiazepin-3-yl] -3-(2,4-dichlorophenyl)prop~anamide

CH

3 0 C1 N; CI

H

To a solution of N-[2,3-dihydro-l1-methyl-2-oxo-5-[2'-(4,4dimethyl-2-oxazolinyl)phenyl]-1 IH-i ,4-benzodiazepin-3 -yl] -3 WO 98/00405 PCTIUS97/11131 -124dichlorophenyl)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 min. the product was extracted into ethyl acetate and purified by column chromatography on silica (60% ethyl acetate/hexane) to give mg N-[2,3-dihydro-1-methyl-2-oxo-[4,5-a]-(1 -oxo-1,3-dihydro-2Hisoindole)-1H-1 ,4-benzodiazepin-3-yl]-3-(2,4-dichlorophenyl)propanamide.

1 H NMR (CDCl3) d: 2.70 2H), 3.12 2H), 3.55 3H), 5.68 (s, 1H), 5.90 1H), 6.85 (dd, 1H), 7.05 (brd, 1H), 7.1-7.5 9H), 7.85 1 H).

MS M+ 1 -494.

EXAMPLE 93

H

3 0 N- N S N H 3R-(+)-3-(Phenylthio)-N-[2,3-dihydro- 1 -methyl-2-oxo-5-phenyl- H- 1,4-benzodiazepin-3-vllpropanamide To a stirred solution of 3-bromopropionic acid in DMF (20 mL) was added K2C03 (1.8 g, 13 mmol) and thiophenol (0.72 g, 6.5 mmol). This was heated to 50 0 C for lh. The mixture was then diluted with 200 mL H20 and extracted with 2 x 100 mL EtOAc. The combined organics were washed with 100 mL and dried with Na2SO4. This was evaporated to give 1.52g of a colorless oil, 1.18g corrected for residual DMF by NMR.

WO 98/00405 PCT/US97/11131 -125 The above oil was taken up in 30 mL DMF and 1-(3dimethylaminopropyl)-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-lmethyl-5-phenyl-2H-1,4-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 H20 and extracted with 2x150mL EtOAc.

The combined organics were washed with 1xl00mL H20, dried with Na2SO04 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 C25H23N302S*0.05Hexane: C, 70.04; H, 5.51; N, 9.69.

Found: C 69.91, H 5.40, N 9.78.

EXAMPLE 94

H

3 C 0 0

HCCH

N *N A S CH 3 N H 3R-(+)-5-(Methylthio)-N-[2,3-dihydro- 1 -methyl-2-oxo-5-phenyl- 1H- 1, 4-benzodiazepin-3-yllpropanamide To an aqueous solution of K2C03 (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 H20 and acidified to pH=O with 6N HCL. Extracted with 2 x 50 mL EtOAc.

Dried with Na2SO4, evaporated to give 0.55g of a yellow oil.

WO 98/00405 PCTIUS97/11131 -126- The above oil was taken up in 10 mL DMF and 1-(3dimethyl-aminopropyl)-3-ethylcarbodiimide hydrochloride 1 6.8mmol) and 1-hydroxybenztriazole hydrate (0.92g, 6.8mmol) were added. 3-(R)-Amino-1,3-dihydro-1-methyl-5-phenyl-2H-1,4benzodaizepin-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 and extracted with 2 x 50 mL EtOAc. Combined organics were dried with brine and Na2SO4, 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 ether:hexane to give a white powder mp. 61-65'C.

Anal. Calcd for C22H25N302S*0.35H20: C, 65.76; H, 6.45; N, 10.46.

Found: C, 65.81; H, 6.21; N, 10.57.

EXAMPLE Me I

N'CN

N

N

-N H H N-cyano-N'-cyclohexylmethyl-N"-(1,3-diliydro- 1 phenvl-2H- 1,4-benzodiazepin-3-vyl)guanidine A solution of 3-(R)-amino-1,3-dihydro-1-methyl-5-phenyl- 2H-1,4-benzodiazepin-2-one (Ig, 3.7 mmole) in acetonitrile (20 mL) was treated with diphenylcyanocarbonimidate (0.9 g, 3.7 mmole) and stirred at room temperature for thirty minutes. Cyclohexylmethyl- WO 98/00405 PCT/US97/11131 127amine (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 HCI 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 Me SN

C

l

H

N-(1,3-Dihydro- -methyl-2-oxo-5-phenyl-2H- 1,4-benzodiazepin-3-yl)- 4-(4-chlorobenzyl)-4-piperidinecarboxamide dihydrochloride Step A: Preparation of N-tert-butyloxycarbonyl-4-(4-chlorobenzyl)-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).

WO 98/00405 PCT/US97/11131 -128- After stirring at -60 0 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 I N HCI 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 6 N HCI (500 mL) and saturated potassium hydrogen sulfate (1 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 0

C,

1 H NMR CDC13 d 7.26 J 8 Hz, 2 7.03 J 8 Hz, 2 H), 3.98 2H), 3.0-2.8 2H), 2.84 2H), 2.10-2.00 m, 2H), 1.55- 1.40 2H), 1.45 9H) Step B: Preparation of N-(1,3-dihydro-1 2H-1,4-benzodiazepin-3-yl)-4-(4-chlorobenzyl)-4-piperidinecarboxamide dihydrochloride A mixture consisting of N-tert-butyloxycarbonyl-4-(4chlorobenzyl)-4-piperidinecarboxylic acid (1.48 g, 4.18 mmole), amino-1,3-dihydro- 1-methyl-5-phenyl-2H- 1,4-benzodiazepin-2-one (1g, 3.7 mmole), hydroxybenzotriazole (1.17 g, 8.66 mmole), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.49 g, 7.70 mmole), diisopropylethyl amine (0.53 g, 4.13 mmole), and DMF mL) was stirred at room temperature for 18 hours. The reaction was WO 98/00405 PCT/US97/11131 129 poured into 1 N HCI 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 HCI 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 C29H29C1N402*2 HCI: 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 Me N O N H NH NH Ci N-(1,3-dihydro- -methyl-2-oxo-5 -phenyl-2H- 1,4-benzodiazepin-3-yl)- 3-(4-chlorobenzyl)-3-piperidinecarboxamide hydrochloride A B isomers WO 98t00405 WO 98100405PCTIUS97/11131 -130- Isomer A m.p. 205 210'C.

Anal. Calcd. for C29H28C1N402-HCI*0.5 CH3CH2OH*0.8 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 C29H28C1N402HC.*O. 1 CH3CH2OCOCH3* 1.6 C, 61.39; H, 5.96; N, 9.74.

Found: C, 61.39; H, 5.66; N, 9.56%.

EXAMPLE 98 NH0

N

N

0r H0 ly0----I 0 WO 98/00405 PCT/US97/11131 131 (+)-3-Cyclohexyl-N-[2,3-dihydro- 1 -methyl-2-oxo-5-phenyl- 1 H-1,4benzodiazepin-3-yll-N-(ethoxycarbonylmethyl)propanamide 3-(R)-Amino- 1,3-dihydro- 1 -methyl-5-phenyl-2H-1,4benzodiazepin-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 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 g of the product.

m.p. 120-122 0 C, [a]D 0.630 (c=0.79; MeOH).

Anal. Calcd. for C29H35N304: 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.

.WO 98/00405 WO 9800405PCTIUS97/11131 132 EXAMPLE 99 3-Cyclohexyl-N-[2,3-dihydro- 1 -methyl-2-oxo-5-phenyl- IH- 1,4benzodiazepin-3-yll -N-(ethoxycarbonylpropvyl)propanamide m.p. 103-105'C, [a]D 0.000; c=0.85; MeOH.

Anal. Calcd. for C31H39N304.*0.40 mol C, 70.94; H, 7.64; N, 8.01.

Found: C, 70.91; H, 7.44; N, 8.12%.

WO 98/00405 WO 9800405PCTIUS97/11131 133 EXAMPLE 100 N. B rr

N

0 N. ."1N N

H

100 N

N

N.

0 N- [2,3 -Dihydro-l1-methyl-2-oxo-5 -phenyl-l1H-I ,4-benzodiazepin-3-yl] N- r2-(2-methoxyethoxy~ethy1hexanmide 3-(R)-Amino- 1,3-dihydro- I-methyl-5-phenyl-2H- 1,4benzodiazepin-2-one (1.33 g, 5.0 inmol) in N,N-dimethyl formamide mL was mixed with 1 -bromo-2-(2-methoxyetho y)ethane (1.35 mL 5.0 mmol) and triethylamnine (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 niL). The organic layers were combined, dried with magnesium sulfate, gravity filtered, and the solvent was removed in WO 98/00405 PCT/US97/11131 -134vacuo. 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 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 an oil, giving 580 mg of the product.

[a]D 0.00°; c=0.27; MeOH.

Anal. Calcd. for C27H35N304.*0.80 mol C, 67.56; H, 7.69; N, 8.75.

Found: C, 67.56; H, 7.39; N, 8.85%.

WO 98/00405 PCT/US97/11131 135 EXAMPLE 101 C 1OH

CI

".IINH

2 0

CI

N

H.,OH

H

0

-N

OH

(+)-N-[2,3-Dihydro- 1-methyl-2-oxo-5-phenyl-1H-1 ,4-benzodiazepin-3- 3-(R)-Amino-1,3-dihydro-1-methyl-5-phenyl-2H-1,4benzodiazepin-2-one (1.33 g, 5.0 mmol) in acetonitrile (40 mL was mixed with 5-chloropentan- 1 -ol (0.61 g, 5.0 mmol) and sodium hydrogen carbonate (2.0 g) was suspended in the mixture. The mixture was stirred 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 a i WO 98/00405 PCT/US97/11131 136solvent was removed in vacuo. The resulting oil was chromatographed over silica in 1:49 methanol:chloroform 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 in vacuo. The resulting oil was chromatographed over silica with 1:1 ethyl acetate:hexane, yielding a foam, giving 360 mg of the product.

foam, [a]d 8.360 (c=0.61, MeOH).

Anal. Calcd. for C27H35N302.*0.25 mol C, 71.42; H, 7.88; N, 9.25.

Found: C, 71.47; H, 7.89; N, 9.12%.

WO 98/00405 PCT/US97/11131 137- EXAMPLE 102 'NCO 2 Et

""IINH

.HNIN CO2E

H

0

CI

-CO

2 Et 0 (+)-N-[2,3-Dihydro- 1-methyl-2-oxo-5-phenyl-1H-1 ,4-benzodiazepin-3yll (ethoxycarbonylpentyl)hexanamide 3-(R)-Amino- 1,3-dihydro- I -methyl-5-phenyl-2H-1,4benzodiazepin-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 stirred 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 WO 98/00405 PCT/US97/11131 138 solvent was removed in vacuo. The resulting oil was chromatographed in 1:49 methanol:chloroform, yielding the mono-alkylated product (0.56 g, 28%) as well as the starting 1,4-benzodiazepin-2-one and bisalkylated 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 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 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 of the product.

m.p. 59-65°C, [a]d (+)52.7o (c=0.48,MeOH).

Anal. Calcd. for C30H39N304.-0.20 mol CH2C12: 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.

WO 98/00405 WO 980405PTIUS97/11131 139 EXAMPLE 103 1Y0-1--* 0 [2,3 -Dihydro- 1 -methyl-2-oxo-5 -phenyl- 1 H-i ,4-benzodiazepin-3 yll -N-(ethoxycarbonylmethl~hexanamide foam, [OC]d 2.04' (c=0.98; MeOH).

Anal. Calcd. for C26H31IN304: C, 69.47; H, 6.95; N, 9.35.

Found: C, 69.41; H, 7.03; N, 9.26%.

EXAMPLE 104 (+)-3-Cyclohexyl-N-[2,3 -dihydro- 1-methyl-2-oxo-5-phenyl- 1H- 1,4benzodiazepin-3-yll -N-(hydroxymethyl)propanamide (+)-3-Cyclohexyl-N-12,3-dihydro- 1 phenyl- 1H-i ,4-benzodiazepin-3 -yllpropanamide (2.0 g, 5.0 mmol) was dissolved in tetrahydrofuran (30 mL), cooled to 0 0 C and methyl WO 98/00405 PCT/US97/11131 140magnesium 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, [a]d 124° (c=0.69, MeOH).

Anal. Calcd. for C26H31N303: C, 72.03; H, 7.21; N, 9.69.

Found: C, 71.66; H, 7.08; N, 9.78%.

The following compound was prepared in a manner substantially as described above starting from (+)-N-[2,3-dihydro-1methyl-2-oxo-5-phenyl-1H- ,4-benzodiazepin-3-yl]hexanamide.

EXAMPLE 105 S00

N

-N

N

\OH

(+)-N-[2,3-Dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin-3yl]-N-(hvdroxvmethyl)hexanamide m.p. 154-156'C, [a]d 190.80 (c=0.24 MeOH).

Anal. Calcd. for C23H27N303*0.30 mol C, 69.26; H, 6.97; N, 10.53.

WO 98/00405 WO 9800405PCTIUS97/1 1131 141 Found: C, 69.29; H, 6.81; N, 10.6%.

EXAMPLE 106 1 0 N

N

'NN

H

(+)-3-Cyclohexyl-N-[2,3 -dihydro-l1-methyl-2-oxo-5-phenyl- 1H- 1,4benzodiazepin-3-yl] -N-(tetrazolylmethyl)propanamide (+)-3-Cyclohexyl-N-[2,3 -dihydro- 1 phenyl- IH-i ,4-benzodiazepin-3-yl] -N-(hydroxymethyl )propanamnide (0.67 g, 1.56 mmol) was dissolved in methylene chloride(100 m.L), along with tetrazole (0.33 g, 4.7 mmol), and then N,N-diisopropyldibenzyl-phosphoramidite (1.07 g, 3.1 mmol). After 2 h, the mixture was diluted with methylene choride (150 mnL), 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, and b (56 mg, Isomer A: m.p. 96-98'C, [oa]d +188.90 (c=0.19, MeOH).

Anal. Calcd. for C27H31IN702-0.30 mol TFA: C, 63.78; H, 6.07; N, 18.86.

Found: C, 63.7; H, 6.12; N, 18.76%.

WO 98/00405 PCT/US97/1131 142- Isomer B: m.p. 92-95 0 C, [a]d +81.30 (c=0.31, MeOH).

Anal. Calcd. for C27H31N7020.35 mol TFA: C, 63.31; H, 6.01; N, 18.66.

Found: C, 63.35; H, 6.02; N, 18.74%.

EXAMPLE 107 00 N "N 0J O N "N

H

3R-(+)-3-(Benzyloxycarbonylamino)-2,3-dihydro- phenyl-1H-1,4-benzodiazepine To a stirring solution of 3-(R)-amino-1,3-dihydro-lmethyl-5-phenyl-2H-1,4-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%) [a]d +57.50 (c=1.17; MeOH).

Anal. Calcd. for C24H20N303.* 0.70 mol H20 -0.15 mol CHC13: C, 67.62; H, 5.06; N, 9.8.

Found: C, 67.6; H, 5.02; N, 9.75%.

WO 98/00405 WO 9800405PCTIVS97/11131 143 The following compounds were prepared substantially as described in Example 81.

EXAMPLE 108 N-[2,3 -Dihydro-l1-methyl-2-oxo-5-ethyl- 1H-I ,4-benzodiazepin-3-yl] -3- (2.4-dichlorophenyl)p2ropanamide m.p. 156-159'C.

CHI-'N: Anal1. C aIc d. fo r C11-H L1 C12N30O2-0-5 H420: 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- 1 -methyl-2-oxo-5-t-butyl- 1 H-i ,4-benzodiazepin-3-yl]- 3-(2,4-dichlorop~henyl)p2ropanamide m.p. 170-171'C.

CH-N: Anal. Calcd. for C23H25C12N302.O.7 C, 60.18; H, 5.80; N, 9.16.

Found: C, 60.17; H, 5.30; N, 9.30.

EXAMPLE N-12,3-Dihydro-l1-methyl-2-oxo[4'-(4,4-dimethyl-2-oxazolinyl)phenyl]- 1 H-i A-benzodiazepin-3-vll -3-(2,4-dichlorop~henvl)propanamide m.p. 188-190'C.

CHIN: Anal. Calcd. for C30H28N403C12: C, 63.95; H, 5.01; N, 9.94.

Found: C, 63.96; H, 5.02; N, 10.08.

EXAMPLE 111 N-[2,3-Dihydro- I -methyl-2-oxo-5-(4-methoxyphenyl)- 1 H- 1,4benzodiazepin-3 -vll -3-(2,4-dichlorophenyl)propanamide WO 98/00405 WO 9800405PCTJIUS97/11131 144 M.P. 189-189'C.

CHN: Anal. Calcd. for C26H23C12N303*O.45 C, 62.91; H, 4.67; N, 8.47.

Found: C, 61.89; H, 4.78; N, 8.33.

EXAMPLE 112 ,5-Dichloro-N-[3R-2,3 -dihydro-2-oxo-5-phenyl-l1-(2,2,2trifluoroethylh- 1H-benzo rel r1.41 diazepin-.3-yilbenzamide.

CF

3 0

CI

I 11N -N H

CI

Step A: Preparation of 2,3-dihydro-2-oxo-5-phenyl-1-(2,2,2trifluoroethyl)- 1H-benzo diazepine.

A solution of 5-phenyl-1 ,4-benzodiazepine-2-one Org. Chem., 1962, 2 7, 3788)(50 g, 0.211 mole) in DMF (100 mL) was treated with cesium carbonate (103.5 g, 0.3 17 mole) and trifluoroethyl iodide (109.7 g, 0.525 mole). The mixture was stirred at overnight. The reaction mixture was then poured into water (2 L) and extracted with ethyl acetate (3 X 1 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 0

C;

I H NMR (CDCI3, 300 MHz) d 7.65-7.60 (in, 2H), 7.60-7.45 (in, 7.40-7.20(m, 2H), 5.25 (dq, J 14, 8.6 Hz, IfH), 4.82(d, J 10.5 Hz, LH), 4.15 (app sextet, J 8.6 Hz, 1H), 3.81 J 10.5 Hz, 1H) WO 98/00405 PCT/US97/11131 -145- Step B: Preparation of 3-Azido-5-phenyl- -(2,2,2-trifluoroethyl)- 1 H-benzo[e] 1,4]diazepine.

To a stirring solution of 5-phenyl-l-(2,2,2-trifluoroethyl)- 1H-benzo[e][1,4]diazepine (70 g,0.22 mol) in THF (1500 mL) cooled to 0 C was added potassium tert-butoxide(1.1 eq, 0.24 mol, 240 mL of a 1 N solution in THF) dropwise over 15 min. A solution of 2,4,6triisopropyibenzenesulfonyiazide (74.8 g, 0.24 mol) in THF (250 ml) was added over 5 min. This was stirred for 10 minutes and acetic acid mL, 0.63 mol) was added and the reaction allowed to warm to ambient temperature. The reaction was poured into satd. NaHCO3 (1500 mL) and ethyl acetate 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 Na2SO4 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( MP 159 160 0

C;

1 H NMR (CDC13, 300 MHz) d 7.70-7.26 5.28-5.12 (m,1H), 4.63 4.35-4.10 (m,lH).

Step C: Preparation of racemic 3-Amino-5-phenyl-l-(2,2,2trifluoroethyl)-1 H-benzo[e] [1,4]diazepine.

To a stirring solution of 3-Azido-2-oxo-5-phenyl-1-(2,2,2trifluoroethyl)-2,3-dihydro-1H-benzo[e][1,4]diazepine (83.4mmol,30g) in 300mL ethanol and 150mL THF was added 10%Pd/C (10 wt%, 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 2 0.0g of white crystals. Another 4g was recovered from evaporation and recrystallization of the filtrates.

Combined yeild, 86.7%.

WO 98/00405 PCT/US97/11131 146- MP 141 143 0

C;

1 H NMR (CDC13,300 MHz) d 7.70-7.26 5.28-5.12 (m,lH), 4.57 4.35-4.10 (m,lH).

Step D: Preparation of 2-Amino-N-[2-oxo-5-phenyl-1-(2,2,2trifluoroethyl)-2,3-dihydro-1 H-benzo[e] 1,4]diazepin- 3-yl]-3-phenylpropionamide To a stirring solution of 3-Amino-2-oxo-5-phenyl-l-(2,2,2trifluoroethyl)-2,3-dihydro-1H-benzo[e][1,4]diazepine (92.2 mmol, 30.74g) in DMF (300mL) was added N-Benzyloxy-D-Phenylalanine (92.2 mmol, 27.6g), 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.12mol,22.95g) and 1-hydroxybenztriazole hydrate (46.1mmol,6.23g). This was stirred at room temperature for 2h. The reaction was then diluted with IL 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 IH (CDC13) d 7.75-7.18 20H), 5.62-5.55 (m,lH), 5.48-5.00 4H), 4.72-4.60 1H), 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-phenyl-l-(2,2,2-trifluoroethyl)-2,3-dihydro-lH-benzo[e][1,4]diazepin- 3-yl]-3-phenyl propionamide in 1L ethanol was added 10% Pd/C 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 WO 98/00405 PCT/US97/11131 -147evaporated 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 18.11g, 83.5% of the upper Rf diastereomer. 1 H NMR (CDCl3,300 MHz) d 8.94 J=8.6Hz, 1H), 7.65-7.10 9H), 5.64 (d, J=8.6 Hz, 1H), 5.28-5.12 1H), 4.57 1H), 4.35-4.10 1H) 3.71 (dd, J=9.8 and 3.9 Hz, 1H), 3.34 dd, J=13.9 and 3.9 Hz,lH), 2.79 (dd, J=13.9 and 10.0 Hz, 1H). The Absolute stereochemistry at C-3 of the benzodiazepine ring was determined to be by X-Ray analysis.

The lower Rf material corresponding to C-3(S) was isolated as well.

Step E: Preparation of 3(R)-(+)-3-Amino-5-phenyl-l-(2,2,2trifluoroethyl)-1 H-benzo[e] diazepine.

To a stirring solution of 2-Amino-N-[2-oxo-5-phenyl-1- (2,2,2-trifluoroethyl)-2,3-dihydro- H-benzo[e][1,4]diazepin-3-yl]-3phenylpropionamide (13.6 g, 28.3 mmol) in methylene chloride (136 mL) was added phenyl isothiocynate (3.87 mL, 34.0 mmol). This was stirred 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: methanol: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 NaHCO3 and 300 mL water. The solution was dried over Na2SO4 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 0

C;

1 H NMR (CDC13,300 MHz) d 7.70-7.26 5.28-5.12 (m,lH), 4.57 4.35-4.10 (m,lH).

WO 98/00405 WO 98/0405 CTIUS91/11 131 148 MaID +72.9' MeOH) The enantiomer wa s 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 5.28-5.12 (m,1H), 4.57 4.35-4.10 (m,1H).

[MD -71.2' (c=0.66, MeOH) Step F: Preparation of ,5-Dichioro-N- [3K-23-di-hydro-2-oxophenyl- 1 -(2,2,2-trifluoroethyl)- 1 H-benzo[e] [1 ,4]diazepin-3 yllbenzamide: To a stirring solution of (+)-3R-3-amino-5-phenyl-1- .(2,2,2-trifluoroethyl)- 1H-benzo [1 diazepmne (5.6 g, 16.8 mmol) in DMF (50 mL) was added 1 -(3-Dimethylaminopropyl-3-ethylcarbodiimide 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 stirred at ambient temperature for 2 hours. The reaction was diluted with 500 mL satd. NaHCO3 and extracted with 2 x 300 mL ethyl acetate. The combined organics were washed with KHS04 (200 mL) brine (200 mL), dried over Na2SO4, and evaporated to a white foam. This was chromatographed over a 75 x 200 mmn silica column eluting with 20% ethyl acetate:hexane. The pure fractions were collected and evaporated under reduced pressure to give 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 0

C,

[a]D=+47.9 0 I H NMR (CDCI3, 300 MHz) d 7.85-7.75 (in, 2 7.70- 7.20 (in, 9 5.78 J=8.1 Hz,l 5.30-5.15 (mn, 1H), 4.30-4.15 (in, 1H) Analysis Calcd. for C24H 1 6053N3 02: C, 56.93; H, 3.19; N, 8.30; Found: C, 56.8 1; H, 3.17; N, 8.17.

WO 98/00405 WO 9800405PCTIUS97/11131 149 The following examples were prepared by a procedure substantially as described for Example 1, Step F.

EXAMPLE 113 ,4-Dichlorophenyl)-N-[ 3R-2,3-dihydro-2-oxo-5-phenyl- I -(2.2.2-trifluoroethyl)- 1H-benzo[ef [1.41 diazepin-3-yllacetamide.

(CF

3 C1 N. NN N

H

mp=219-221 0 C; [a]D=-10.8 0 IH NMR (CDCl3,300 MHz) d 7.65-7.15 (in, 12H), 5.78 1=8.1 Hz, 1H), 5.25-5.10 (in, LH), 4.25-4.05 (in, 3.56 2H); Analysis Calcd. for C25H18SC12F3N302*0.85 C, 56.06; H, 3.7 1; N, 7.84.

Found: C, 56.03; H, 3.53; N, 7.82.

EXAMPLE 114 ,5-Dichlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5 -phenyl- 1 -(-2.2.2-trifluoroethyl)- 1H-benzo[elf [1.4]diazepin-3-vil acetamide WO 98/00405 WO 9800405PCTIUS97/11 131 150 F3 mp=93-100 0 C, [MD -5.70; 1H NMR (CDCl3,300 MHz) d 7.65-7.15 (in, 12H), 5.78 J=8.1 Hz, 1H), 5.25-5.10 (in, 1H), 4.25-4.05 (in, 1H), 3.65 2H); Analysis Calcd. for C25H18C12F3N302: C, 57.7 1; H, 3.49; N, 8.08; Found: C, 57.41; H, 3.48; N, 8.12.

EXAMPLE 115 ,5-Bis(trifluoromethyl)phenyl] [3R-2,3 -dihydro-2-oxo- 5-phenyl-lI-(2,2,2-trifluoroethyl)- 1H-benzo [1 ,4]diazepin-3 villacetamide (F3 qF 3 m.p. foam [MD -9.70 (c=0.59,MeOH).

Anal. Caled. for C27H1I8F9N302*0.75 C, 53.96; H, 3.27; N, 6.99.

WO 99/00405 WO 9800405PCTIUS97/11131 151 Found: C, 53.96; H, 3.1; N, 6.9R%.

EXAMPLE 116 (-)-2-(4-Trifluoromethylphenyl)-N-[3R-2,3 p2henyl- 1 -(2.2.2-trifluoroethyl)- 1 H-benzore] 1 .ldiazepin-3-yllacetamnide

CF

3 m.p. 253-255 Ml)] (c=0.25, MeOH).

Anal. Calcd. for C26HI19F6N3O2-0.05 ethyl etherO0.55 C, 59.03; H, 3.9; N, 7.88.

Found: C, 59.05; H, 3.82; N, 7.78%.

EXAMPLE 117 2-(3 -Trifluoromethylphenyl)-N- r3R-2, 3-dihydro-2-oxo-5 -phenyl 1 -(2.2,2-trifluoroethyl)- 1H-benzo[el r1.41 diazepin-3-yll acetamide

CF

3 N H m.p. 172-173 [l]Dl (c=0.56, CHC13).

WO 98/00405 WO 9800405PCTIUS97/1 1131 152 Anal. C alcd. for C26H1I9F6N302O.60 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 phenvl-l1-(2 .2.2-trifluoroethyl)- 1H-benzo[el [1 Al diazepin-3-yl acetamide

CF

3 H CF 3 m.p. 170-17 1 Ml)D +9.00 (c=0.48, CHCI3).

Anal. Caled. for C26H I9F6N302*0.25 C, 59.6; H, 3.75; N, 8.02.

Found: C, 59.64; H, 3.68; N, 7.97%.

EXAMPLE 119 (-)-2-(2,4-Dichlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5-phenyl- 1 -(2.2..2-trifluoroethyl)- 1H-benzo[el [1.41 diazep~in-3-yll acetamide

F

3 C WO 98/00405 WO 9800405PCTIUS97/11131 153 m.p. 143-145 [MD -22.6' (c=0.73; MeOH).

Anal. Calcd. for C25H1I 8N3 02C12F3: C, 57.71; H, 3.49; N, 8.08.

Found: C, 57.75; H, 3.52; N, 8.09%.

EXAMPLE 120 -)--3-Chlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5'-phenyl- 1 (2.2.2-trifluoroethyl)- 1H-benzo [el [1.41diazev~in- 3-vi] acetamide

(CF

3 m.p. 188-189 NOlD 4' (c=1.03,MeOH).

Anal. Calcd. for C25H1I9CIF3N3020. 10 ethyl ether: C, 61.84; H, 4.09; N, 8.52.

Found: C, 61.84; H, 4.05; N, EXAMPLE 121 (-)-2-(4-Chlorophenyl)-N- L3R-2,3 14 (222-trifluoroethyl)- 1H-benzore] [1.41diazepin-3-yll acetamide WO 98/00405 WO 9800405PCTfUS97/11131 154-

CF

3

C

m.p. 246-247 [ax]D Anal. Caled. for C25H I9C1F3N302O.20 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] [3R-2,3-dihydro-2-oxo-5p2henyl-l1-(2 .2,2-trifluoroethyl IH-benzo[el f1.41 diazepin-3 -vl acetamide

CF

3

-CF

3 N

H

CF

3 Step A. 2,4-Bis(trifluoromethyl)benzonitrile To a stirring biphasic mixture of lO0mL- ethanol and 250 mL of phosphate buffer (1 g of NaH2PO4'H20 per 5 mL H20 adjusted to pH=7.O with 50% NaOH) and NaCN (81 .3mmrol,4.Og) heated to was added 2,4-bis(trifluoromethyl) benzyl bromide (3 2 .5mmol,l0g) in EtOH dropwise over 30min. The reaction was heated at 60 0

C

WO 98/00405 PCT/US97/11131 155 for 24h. The reaction was then evaporated under reduced pressure.

The remaining aqueous was extracted with 2x150mL EtOAc. The organic layers were combined, dried with brine and Na2SO4. 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 IH (CDC13) d 8.0-7.85 4.03 (s,2H) Step B. 2,4-Bis(trifluoromethyl)phenyi acetic acid 2,4-Bis(trifluoromethyl)benzonitrile (41.5mmol, 10.51g) was taken up in 100mL acetic acid, 50mL conc. H2S04, and water. This was heated to 120 0 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 Na2SO4, 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(trifluoromethyl) phenyl acetic acid as white crystals, 68.5%.NMR 1 H (CDC13) d 7.93 7.80 J=7.9Hz,1H), 7.55 J=7.9Hz,H), 3.94 (s,2H).

Step C. Preparation of (-)-2-[2,4-Bis(trifluoromethyl)phenyl]-N- [3R-2,3-dihydro-2-oxo-5-phenyl- -(2,2,2-trifluoroethyl)- 1H-benzo[e] [1,4]diazepin-3-yl]acetamide To a stirring solution the 3R-3-Amino-2-oxo-5-phenyl-1- (2,2,2-trifluoroethyl)-2,3-dihydro-1 H-benzo[e] [1,4]diazepine (28.4 mmol, 9.47g) in DMF (100mL) was added 2,4-Bis(trifluoromethyl) phenyl acetic acid (28.4mmol,7.74g), 1-(3-Dimethylaminopropyl)-3ethylcarbodiimide hydrochloride (42.6mmol,8.16g) and 1-Hydroxybenztriazole hydrate (14.2mmol,1.92g). This was stirred for Ih at room temperature. The reaction was then diluted with 750mL of KHSO4 and extracted with ethyl acetate (2x300mL). The organic layers WO 98/00405 PCT/US97/11131 -156 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 0 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 crystallinity.

NMR 1 H (CDC13) d 7.95-7.25 (m,13H), 5.60 (d,J=8.1Hz,lH), 5.30- 5.10 4.25-4.06 3.96 (s,2H) Anal. Calcd. for C27H18F9N302: 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 -(2,22-trifluoroethyl)- H-benzo [el 141diazepin-3-vyl acetamide o RC N I N CI N

H

m.p. 219-220 racemic Anal. Calcd. for C25H 18N302C12F3: C, 57.71; H, 3.49; N, 8.08.

Found: C, 57.94; H, 3.48; N, 8.02%.

WO 98/00405 WO 9800405PCT/US97/11131 157 EXAMPLE 124 2-(3 ,5-dichloro-4-methoxyphenyl)-N-[3R-2,3-dihydro-2-oxo-5 phenyl-l1-(2,2,2-trifluoroethyl)- 1H-benzore] [1,41diazepin-3 -viiacetamide m.p. 100-104 RX]D -8.90 (c=0.55,MeOH).

Anal. Calcd. for C26H20C12F3N 303: 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 I except substituting the appropriate fluoro substituted aminobenzophenone in step A.

EXAMPLE 125 ,5 -Dichlorophenyl)-N- [2,3 -dihydro-5 -(4-fluorophenyl)-2oxo-l1-(2.2.2-trifluoroethyl)- 1H-benzo[el [1.41diazein-3-yl] acetamide WO 98/00405 WO 9800405PCT/US97/11131 158

F

3 C m.p. foam [MD (c=0.55; MeOH).

Anal. Calcd. for C25H17N302C12F4: C, 55.78; H, 3.18; N, 7.81.

Found: C, 55.73; H, 3.25; N, 7.72%.

EXAMPLE 126 (2,4-Dichlorophenyl)-N- [2,3-dihydro-5 -(4-fluorophenyl)-2oxo-l1-(2.2.2-trifluoroethyl)- IH-benzo rel r1 ,4ldiazepin-3-vll acetamide m.p. foam [ax]D =-I1I' (c=0.68; MeOH).

Anal. Calcd. for C25H17N302F4: C, 55.78; H, 3.18; N, 7.81.

Found: C, 55.82; H, 3.41; N, 7.42%.

WO 98/00405 WO 9800405PCTIUS97/11131 159 EXAMPLE 127 ,5-Bis(trifluoromethyl)phenyl)-N- [2,3-dihydro-5-(4fluorophenyl)-2-oxo-l1-(2,2,2-trifluoroethyl)- 1H-benzo[e] f1 .4]diazepin-3-yl]-acetamide

PF

3

F

3 C

CF

3 m.p. foam 0 C, [a]D +2.80 (c=0.67; MeOH).

Anal. Calcd. for C27H17N302FI0: C, 53.56; H, 2.83; N, 6.94.

Found: C, 53.56; H, 2.93; N, 6.91 EXAMPLE 128 [2,4-Bis(trifluoromethyl)phenyl] [2,3-dihydro--5 fluorophenyl)-2-oxol1-(2,2,2-trifluoroethyl)- 1H-benzo [1 ,4] diazepin-3-y1 acetamide WO 98/00405 WO 9800405PCTIUS97/11131 160 F3CG>,

CF

3

CF

3 Ml)D 14' (c=0.63; MeOH).

Anal. Calcd. for C27H17N302F10: 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- 1H-benzo rel [141 diazepin -3-ylllpropionamide

N

IH

m.p. 202-204 'C.

I H NMR d (CDCl3) 7.72 5.65 (d,J=8.3Hz,1H), 5.35-5.08 4.32-4.15 2.37 (t,J=7.8Hz,2H), 1.80-1.55 (m,711), 1 .45-Anal. Calcd. for C26H27F4N302: C, 63.8; H, 5.56; N, 8.58.

WO 98/00405 WO 9800405PCTIUS97/11131 161 Found: C, 63.82; H, 5.54; N, 8.56%.

EXAMPLE 130 3 ,4-Dichloro-N-[2,3-dihydro-5 -(2-fluorophenyl)-2-oxo- 1- (2.2 .2-trifluoroethyl)-l1H-benzo re][1.41 diazepin-3-yllbenzamide

CF

3

~CI

N H

C

F

m.p. 168-170 'C.

IH NMR d (CDC13) 8.03 7.86 (d,J=7.8Hz,1H), 7.78-7.05 5.80 (d,J=7.8Hz,1H), 5.27-5.15 4.35-4.20 (m,1H) Anal. Calcd. for C24H 15C0504302: C, 54.98; H, 2.88; N, 8.01.

Found: C, 54.96; H, 2.89; N, 8.12%.

EXAMPLE 131 Antiarrhythmic efficacy of the combined administration of low dose IKs blocker (-)-2-[2,4-bis(trifluoromethyl)phenyl]-n-[3r-2,3dihydro-2-oxo-5-phenyl-lI-(2,2,2-trifluoroethyl)- 1h-benzo [1 ,4] diazepin-3-yllacetamide and the beta-adrenergic receptor antagonist timolol in anesthetized dogs with previous anterior myocardial infarction was studied. This study was conducted to test the hypothesis the concomitant adminis-tration of low dose lKs blocker Bis(trifluoromethyl)phenyl] [3R-2,3-dihydro-2-oxo-5 -phenyl- 1- (2,2,2-trifluoroethyl)- 1H-benzo [1 diazepin- 3-yI] acetamnide and WO 98/00405 PCT/US97/11131 162 low dose beta-adrenergic receptor antagonist timolol, which when administered individually at low dose afford no significant antiarrhythmic protection, would in combination provide significant protection against the development of malignant ventricular arrhythmias 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 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 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 WO 98/00405 PCT/US97/11131 163 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 extrastimuli 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 extrastimuli during sinus rhythm and atrial pacing was performed at the RVOT site. If ventricular extrastimuli could not be inserted at the RVOT site, programmed pacing was attempted at the IZ site.

Ventricular extrastimuli were introduced at 2x diastolic threshold voltage or, if extrastimuli were not introduced adequately, at 4x diastolic threshold voltage. Responses to baseline programmed ventricular stimulation were categorized as: 1) non-inducible less than 5 nonstimulated ventricular complexes; 2) nonsustained ventricular tachycardia (NSVT): 5 or more nonstimulated ventricular complexes terminating spontaneously with a duration less than seconds; 3) unimorphic sustained ventricular tachycardia (UVT): unimorphic ventricular tachycardia with a duration exceeding seconds; 4) polymorphic sustained ventricular tachycardia (PVT): WO 98/00405 PCT/US97/11131 164polymorphic ventricular tachycardia with a duration exceeding 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 tachyarrhythmias (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 arrhythmic 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 current of 200 pA 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 electrocardiographically. Upon the development of lethal ischemic arrhythmias 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 WO 98/00405 PCT/US97/11131 165 quantitated gravimetrically and was expressed as a percentage of total left ventricle.

The responses occurrence of vs protection from the development of lethal ventricular arrhythmia) 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 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 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- 1 -(2,2,2-trifluoroethyl)- Hbenzo[e][ 1,4]diazepin-3-yl]acetamide in soy bean oil-based microemulsion 4) low dose 0.001 mg/kg i.v. beta-adrenergic receptor blocking agent timolol in 5% dextrose in saline vehicle and 5) the combination of low dose 0.0003 mg/kg i.v. IKs blocker (-)-2-[2,4-Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5phenyl-1-(2,2,2-trifluoroethyl)-1H-benzo[e] [1,4]diazepin-3-yl]acetamide with low dose 0.001 mg/kg i.v. beta-adrenergic receptor blocking agent timolol In the individual and combination IKs blocker [2,4-Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5-phenyl-1- (2,2,2-trifluoroethyl)-1H-benzo[e][1,4]diazepin-3-yl]acetamide and betaadrenergic 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.

WO 98/00405 PCT/US97/11131 166-

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 arrhythmias (ventricular fibrillation; VF) occurring in the laboratory control cohort, dedicated microemulsion vehicle control group, low dose IKs blocker Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5-phenyl- 1- (2,2,2-trifluoroethyl)- 1 H-benzo[e][1 i diazepin-3 -yljacetamide, 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)-1H-benzo[e][1,4]diazepin-3yl]acetamide with beta-adrenergic receptor blocking agent timolol groups are summarized below.

P Value Incidents of Lethal P Value Compared Compared to Ischemic to Laboratory Microemulsion Ventricular Vehicle Control Vehicle Control Treatment Group Arrhythmic (VF) Cohort Group Laboratory Vehicle 34/40 NS Control Cohort Microemulsion 8/10 NS Vehicle Control Group Low Dose IKs 6/10 NS NS Blocker 0.0003 mg/kg i.v.

Low Dose B Blocker 4/6 NS NS 0.001 mg/kg i.v.

Timolol Combined Low Dose 3/10 0.001 0.03 IKs Blocker 0.0003 mg/kg i.v. Low Dose B Blocker 0.001 mg/kg i.v.

Timolol NS nonsignificant (P>0.05).

WO 98/00405 PCT/US97/11131 167 Neither low dose IKs blocker Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5-phenyl-1- (2,2,2-trifluoroethyl)- H-benzo[e][1,4]diazepin-3-yl]acetamide nor low dose beta-adrenergic receptor antagonist timolol administered individually afforded significant protection against the development of malignant ischemic ventricular arrhythmias 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 IKs blocker Bis(trifluoromethyl)phenyl]-N-[3R-2,3-dihydro-2-oxo-5-phenyl-1- (2,2,2-trifluoroethyl)- H-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 tachyarrhythmia and arrhythmic death, which signifies a synergistic salutary antiarrhythmic interaction between block of IKs and betaadrenergic receptor blockade.

Claims (3)

169- Y is 1) =0, 2) N-CN or 3) H2; Z is 1) C1-6 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 or -NH, 4) 4-(5-methylisoxazole-3-yl), C3-6 cycloalkylene, or 6) single bond; p is 0 or 1; R 1 is 1) phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02, b) -Cl, Br, F, or I, c) -CF3, d) -C1-3 alkyl, e) -C1-3 alkoxy, f) -CN, g) -methylenedioxy, 2) C5-7 cycloalkyl, 3) CO 2 t-Bu, 4) mono- or bicyclic heterocyclyl of 5 to 10 members one or two of which are sulfur, nitrogen or oxygen, the remaining WO 98/00405 PCT/US97/11131

170- being carbon, such as 2-thienyl, 2-furanyl, 2-indolyl, 2- quinoxolinyl, or 2-(2,3-dihydro benzofuranyl) methyl, or 6) R 2 is 1) phenyl, either unsubstituted or substituted with CI-3 alkoxy or 4,4-dimethyloxazolin-2-yl, 2) C1-4 alkyl, either straight or branched chain and either unsubstituted or substituted with CI-3 alkoxy or C1-3 alkoxy-C1-3 alkoxy, 3) C5-7 cycloalkyl, 4) 2- or 3-furyl, 1 -methylpiperidin-2-yl, or 6) if R 2 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 3 is 1) hydrogen or 2) C1-6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3, or 3) -CF3; R 4 is 1) hydrogen, 2) C1-6 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 C1-3 alkoxycarbonyl, -OH or N0 2 or 3) tetrazol-5-yl; and 171 R 5 is hydrogen or oxygen or is joined structure: to R2 to form the partial and the bond represented by is: 1) a double bond when p is zero or when p is 1 and R 5 is oxygen, or 2) a single bond when R 5 is hydrogen or R 5 is joined to R2 to form the partial structure: 0 9 0 *9 .9 2. The selective IKs compound of Claim I wherein A is benzo either unsubstituted or substituted with -NH2, -NHSO2(C1-3 alkyl), C1-3 alkyl or C1-3 alkoxy; X and Y are 0, Z is 1) C1-6 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 or -NH, 4) 4-(5-methylisoxazole-3-yl), C3-6 cycloalkylene, or WO 98/00405 WO 9800405PCTIUS97/1 1131 172 6) single bond; RI is 1) phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02, b) -Cl, Br, F, or 1, c) -CF, d) -C1-3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy, 2) C5-7 cycloalkyl, 3) CO 2 t-Bu, 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) methyl, or 6) R 2 is 1) phenyl, either unsubstituted or substituted with Ci 1.3 alkoxy or 4,4-dimethyloxazolin-2-yl, 2) Ci1 -4 alkyl, either straight or branched chain and either unsubstituted or substituted with C 1-3 alkoxy or C1-3 alkoxy-Cl-3 alkoxy, 3) C5-7 cycloalkyl, 4) 2- or 3-furyl, 1 -methylpiperidin-2-yl, or WO 98/00405 PCT/US97/11131

173- 6) if R 2 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 3 is 1) hydrogen or 2) C -6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3, or 3) -CF3; R 4 is 1) hydrogen, 2) C -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 CI-3 alkoxycarbonyl, -OH or NO 2 or 3) tetrazol-5-yl; and R 5 is hydrogen or oxygen or is joined to R 2 to form the partial structure: N 0 and the bond represented by is: 1) a double bond when p is zero or when p is 1 and R 5 is oxygen, or 2) a single bond when R 5 is hydrogen or R 5 is joined to R 2 to form the partial structure: -174- 0 including individual diastereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt. 3. The selective IKs compound of Claim 2 wherein A is benzo either unsubstituted or substituted with -NH2 -NHSO2(C1-3 alkyl), C1-3 alkyl or C1-3 alkoxy; X and Y are 0, Z is 1) C1-6 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 or -NH, 4) 4-(5-methylisoxazole-3-yl), 5) C3-6 cycloalkylene, or 6) single bond; R1 is phenyl, either unsubstituted or substituted with one or two substituents selected from a) -N02, b) -Cl, Br, F, or I, c) -CF3, d) -C1-3 alkyl, e) -Cl-3 alkoxy, f) -CN, g) -methylenedioxy, 175 R 2 is phenyl, either unsubstituted or substituted with C 1-3 alkoxy or 4,4-dimethyloxazolin-2-yl, R 3 is -CF3 or C 1-6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3; R 4 and R 5 are hydrogen; inicluding individual diasrereomers, enantiomers and mixtures thereof, or a pharmaceutically acceptable salt. 4. The selective IKs corrpound of Claim 3 wherein A is unsubstituted benzo; *0 0@ I 0@ S 0*O@ *0 9 @0 0 *006 *500 S 4 S 0S55 .9 4 0 9 0@ SW S 0 0 5 9 0 055. S 0 S0 9 S 090 0 0**0 a 0 *WS* 0 0* S S 0S*000 0 RI is phenyl, either unsubstiruted or substituents selected from a) -N0O, b) -Cl1. B r. F. o r L, c) -CF3, e) -C1-I alkoxy, f) -CN, a) -methvlenedioxV. substituted with one or two R 2 is phenyl, either unsubstituted or substituted with Cl_-3 aikoxy or 4 ,4-dimethyloxazolin-2-yl, R 3 is -CF3 or C 1-6 alkyl, either straight or branched chain and either unsubstituted or substituted with -N(CH3)2, -OH, -CF3; R 4 and R 5 are hydrogen; including individual diastereomers, enantiomers and m~ixtures thereof, or a pharmaceutically acceptable salt. 176 The selective IKs compound of Claim 4 which is 2-[2,4-Bis(trifluoromethyl)phenyl] -N-Ii3R-2,3-dihydro-2-oxo-5-phenyl- 1 -(2,2,2-trifluoroethyl 1 H-benzo 1,4] diazepin-3-yl] acetamide (CF 3 -CF 3 9 9* 9. 9 6. The selective IKs compound of Claim 4 which is 3,4- Dichloro-N- [2,3 -dihydro -5 -(2-fluo ropheny1) oxo -I -(2,2,2-tri flu oro ethyl)-1IH-benzo[e][ 1 ,4]diazepin-3-yl]benzamide 7. The selective IKs compound of Claim 4 which is (-)-2-(2,4-Dichlorophenyl)-N-[3R-2,3-dihydro-2-oxo-5-phenyl- I (2,2,2-trifluoroethyl)- I H-benzo[e] 1,4] diazepin-3-yl]acetamide 177 F 3 C O 0 Cl N H N Hl N CI 8. A selective IKS compound which is substantially as hereinbefore described with reference to the title compound in any one of the Examples. 9. A pharmaceutical formulation comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a beta-adrenergic receptor blocking agent and a selective IKs antagonist. The pharmaceutical formulation of claim 9 wherein the selective IKS antagonist is the compound of any one of claims 1 to 8. 11. The pharmaceutical formulation of claim 9 or claim 10 wherein the beta- adrenergic blocking agent is selected from the group consisting of timolol, sotalol, esmolol, cateolol, propranolol, betaxolol, penbutolol, metoprolol, acebutolol, atenolol, metoprolol, pindolol and bisoprolol. 12. A method of treating or preventing arrhythmia, which method comprises the co- administration to a patient of an effective amount of a beta-adrenergic receptor blocking agent and a selective IKS antagonist. 13. The method of claim 11 wherein the selective IKS antagonist is a compound of any one of claims 1 to 8. 14. The method of claim 12 or claim 13 wherein the beta-adrenergic receptor blocking agent is selected from the group consisting of timolol, sotalol, esmolol, cateolol, propranolol, betaxolol, penbutolol, metoprolol, acebutolol, atenolol, metoprolol, pindolol and bisoprolol. A method of treating or preventing arrhythmia, which method comprises the administration to a patient of an effective amount of the pharmaceutical formulation of any one of claims 9 to 11. 16. A beta-adrenergic receptor blocking agent and a selective IKS antagonist when used to treat or prevent arrhythmia. 17. Use of a beta-adrenergic receptor blocking agent and a selective IKS antagonist m the manufacture of a medicament to treat or prevent arrhythmia. [R:\LIBAA]08542speci.doc:tIt 178 18. A compound of any one of claims 1 to 8 and a beta-adrenergic receptor blocking agent when used to treat or prevent arrhythmia. 19. Use of a compound of any one of claims 1 to 8 and a beta-adrenergic receptor blocking agent in the manufacture of a medicament to treat or prevent arrhythmia. 20. A compound of any one of claims 1 to 8 and a beta-adrenergic receptor blocking agent selected from the group consisting of timolol, sotalol, esmolol, cateolol, propranolol, betaxolol, penbutolol, metoprolol, acebutolol, atenolol, metoprolol, pindolol and bisoprolol when used to treat or prevent arrhythmia. 21. Use of a compound of any one of claims 1 to 8 and a beta-adrenergic receptor blocking agent selected from the group consisting of timolol, sotalol, esmolol, cateolol, propranolol, betaxolol, penbutolol, metoprolol, acebutolol, atenolol, metoprolol, pindolol and bisoprolol in the manufacture of a medicament to treat or prevent arrhythmia. 22. A pharmaceutical formulation of any one of claims 9 to 11 when used to treat or prevent arrhythmia. S 15 Dated 10 May, 2000 Merck Co., Inc. Patent Attorneys for the Applicant/Nominated Person SPRUSON FERGUSON o *oo o*o 0•0 o*o *•o [R:\LIBAA]08542spci.doc:tlt