AU714287B2 - Dihydropyrimidines and uses thereof - Google Patents

Description

WO 97/17969 PCT/US96/18573 DihYdropyrimidines and Uses Thereof This application is a continuation-in-part of PCT International Application No. PCT/US95/15025, filed .0 November 16, 1995, the contents of which are incorporated by reference. Throughout this application, various references are referred to within parentheses. Disclosures of these publications in their entireties are hereby incorporated by reference into this application to more fully describe the state of the art to which this invention pertains.

Backaround of the Invention The designation "au" is the appellation recently approved by the IUPHAR Nomenclature Committee for the previously designated "a c" cloned subtype as outlined in the 1995 Receptor and Ion Channel Nomenclature Supplement (Watson and Girdlestone, 1995). The designation ajA is used throughout this application and the supporting tables and figures to refer to this receptor subtype. At the same time, the receptor frr designated a1A was renamed The new nomenclature is used throughout this application.

Stable cell lines expressing these receptors are described herein; however, these cell lines were deposited with the American Type Culture Collectlon (ATCC) under the old nomenclature (infr).

Benign Prostatic Hyperplasia (BPH), also called Benign Prostatic Hypertrophy, is a progressive condition which is characterized by a nodular enlargement of prostatic SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 :tssue resulting in obstructin of the urethra. This results in increased frequency of urination, nocturia, a poor urine stream and hesitancy or delay in start-ng the urine flow. Chronic consequences of BPH can include hypertrophy of bladder smooth muscle, a decompensated bladder and an increased incidence of urinary tract infection. The specific biochemical, histological and pharmacological properties of the prostate adenoma leading to the bladder outlet obstruction are not yet known. However, the development of BPH is considered to be an inescapable phenomenon for the aging male population. BPH is observed in approximately 70% of males over the age of Currently, in the United States, the method of choice for treating BPH is surgery (Lepor, Urol.

Clinics North Amer., 17, 651 (1990)). Over 400,000 prostatectomies are performed annually (data from 1986). A medicinal alternative to surgery is clearly very desirable. The limitations of surgery for treating BPH include the morbidity rate of an operative procedure in elderly men, persistence or recurrence of obstructive and irritative symptoms, as well as the significant cost of surgery.

ac-Adrenergic receptors (McGrath, et. al. Med. Res.

Rev., 9, 407-533, 1989) are specific neuroreceptor proteins located in the peripheral and central nervous systems on tissues and organs throughout the body.

These receptors are important switches for controlling many physiological functions and, thus, represent important targets for drug development. In fact, many a-adrenergic drugs have been developed over the past years. Examples include clonidine, phenoxybenzamine and prazosin (treatment of hypertension), naphazoline (nasal decongestant), and apraclonidine (treating glaucoma). a-Adrenergic drugs can be broken down into two distinct classes: agonists (clonidine and SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 naphazoline are agonists which mimic the receptor activation properties of the endogenous neurotransmitter norepinephrine, and antagonists (phenoxybenzamine and prazosin are antagonists), which act to block the effects of norepinephrine. Many of these drugs are effective but also produce unwanted side effects (for example, clonidine produces dry mouth and sedation in addition to its antihypertensive effects).

During the past 15 years a more precise understanding of a-adrenergic receptors and their drugs has evolved through increased scientific scrutiny. Prior to 1977, only one a-adrenergic receptor was known to exist.

Between 1977 and 1988, it was accepted by the scientific community that at least two a-adrenergic receptors--a, and a,--existed in the central and peripheral nervous systems. Since 1988, new techniques in molecular biology have led to the identification of at least six a-adrenergic receptors which exist throughout the central and peripheral nervous systems: aIA (new nomenclature), a (new nomenclature), a2, and a 2 c (Bylund, FASEB 6, 832 (1992)). In many cases, it is not known precisely which physiological responses in the body are controlled by each of these receptors. In addition, current a-adrenergic drugs are not selective for any particular a-adrenergic receptor. Many of these drugs produce untoward side effects which may be attributed to their poor a-adrenergic receptor selectivity.

Since the mid 1970's, nonselective a-antagonists have been prescribed to treat BPH. In 1976, M. Caine, et al. (Brit. J. Urol., 48, 255 (1976)), reported that the nonselective a-antagonist phenoxybenzamine was useful in relieving the symptoms of BPH. This drug may produce its effects by interacting with a-receptors SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -4located on the prostate. However, :his drug also produces significant side effects such as dizziness and asthenia which severely limit its use in treating patients on a chronic basis. More recently, the a-adrenergic antagonists prazosin and terazosin have also been found to be useful for treating

BPH.

However, these drugs also produce untoward side effects. It has recently been discovered that the a A receptor is responsible for mediating the contraction of human prostate smooth muscle (Gluchowski, C. et.

al., WO 94/10989, 1994; Forray, C. et. al., Mol.

Pharmaco. 45, 703, 1994). This discovery indicates that the a1A antagonists may be effective agents for the treatment of BPH with decreased side effects. Further studies have indicated that the al receptor may also be present in other lower urinary tract tissues, such as urethral smooth muscle (Ford et al. Br. J. Pharmacol., 114, 24P, (1995)).

This invention is directed to dihydropyrimidine compounds which are selective antagonists for cloned human ao receptors. This invention is also related to uses of these compounds for lowering intraocular pressure (Zhan, et. al. Ophthalmol. Vis. Sci., 34 Abst.

#1133, 928, 1993), inhibiting cholesterol synthesis (D'Eletto and Javitt, J. Cardiovascular Pharmacol., 13 (Suppl. 2) S1-S4, 1989), benign prostatic hyperplasia, impotency (Milne and Wyllie, EP 0 459 666 A2, 1991), sympathetically mediated pain (Campbell, WO 92/14453, 1992), cardiac arrhythmia (Spiers, et. al.,J.

Cardiovascular Pharmacol., 16, 824-830, 1990) and for the treatment of any disease where antagonism of the acA receptor may be useful.

SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 Summary of the Invention This invention is directed to dihydrcpyrimidine compounds which are selective antagonists for human a, receptors. This invention is also related to uses of these compounds for lowering intraocular pressure, inhibiting cholesterol synthesis, relaxing lower urinary tract tissue, the treatment of benign prostatic hyperplasia, impotency, cardiac arrhythmia and for the treatment of any disease where antagonism of the a, receptor may be useful. The invention further provides a pharmaceutical composition comprising a therapeutically effective amount of the above-defined compounds and a pharmaceutically acceptable carrier.

SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 97/ 7969PCT/US96/18573 Detailed Description of the Invention The uresent invention is directed to compounshvn the structures:

M

21 R 3 R9\ x N N R 3 SUB8STITUTE S H EET (RU LE 26) WO 97/17969 WO 9/1 969PCT/UJS96/18573

N

N

where M has the structure wherein each of Y1, Y 2

Y

3 Y, and Ys independently may be straight chained or branched alkyl, monofluoroalky. or polyf luoroalkyl; straight chained or branched

C

2

-C

7 alkeny. or alkynyl;

C

3 cycloalkyl, monofluorocycloalkyl polyfluorocycloalkyl or cycloalkenyl; -Cl, -Br, or

-NO

2 -N (R 3 2

-N

3 -CN;

-OR

3 -OCOR 3

-COR

3

-CON(R

3 2 or -CO,R 3 or wherein two of Y 1 Y21

Y

3

Y

4 and Y, are present on adj a cent carbon atoms and together constitute a methylenedioxy group; SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 9/1 969PCT/tJS96/18573 where R independently may be Straight chained or branched C, alkyj, monofJluoroalkyl Cor polyf luoroalkyl; straight chained or branche~d C C.

alkeny. or alkynyl; -N(C,3) 2

-NC

2 -CN; -2 2 aR 3 (CH 2 O(0R 3

N/

2

-(CH

2 PO(OR3) 3; or -OR 3 where R, may be -NO 2 -CN; straight chained or branched Cl-C? alkyl, monofluoroalkyl or polyf luoroalkyl; straight chained or branched C2 C 7 alkenyl or alkynyl;

C

3 cycloalkyl, monofluorocycloalkyl, polyfluorocyclAoalkyli or cycloalkenyl;

-NC(R

3 2

-OR

3

-CCH

2

POR

3

-COR,;

-C0 2

R

3 or -CON(R 3 ,2, wherein any p independently is an integer from 1 to 7 inclusive; where R 2 may be straight chained or branched C,-C.,alkyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched

C

2 alkenyl or alkynyl;

C

3

-C,

cycloalkyl, monofluorocycloalkyl, polyfluorocycloalkyl or cycloalkenyl;

C

3

-C

13 cycloalkyl-C,-Cl 0 -alkyl, C3-C 10 cycloalkyl-C,-C, 1 0 monofluoroalkyl or C 3 -CIO cycloalkyl-C,-C, 1 0 polyfluoroalkyl; -CN; CH 2

XR

3

-CH

2 X (CH 2

PNMR

3

(CH

2

JNH

3

-CH

2 X CCH 2 P N CR 3 2' -CI 2 X (CE 2

AN

3 or

_CH

2

X(CH

2 )pNNCXR,; or -OR 3 wherein any n independently is an integer from 0 to 5 inclusive and p is as defined above; where each R 3 independently may be straight chained or branched alkyl, monofluoroalkyl or polyf luoroalkyl; straight chained or branched

C,-C,

alkenyl or alkynyl;

C

3 cycloalkyl, monofluorocycloalkyl, polyfluorocycloalkyl or SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCTIUS96/18573 -9cycloalkenyl;

-(CH

2 )PPO(OR 8 2

-(CH

2 PO(OR)3 Or -(CH 2 Ca7R S where R 4 has the structure

R

R R 3 R 6

RR

m wherein Z may be C,-C alkenyl or alkynyl; CH2; 0; CO; CO 2 CONR3CO; CONR 3 S; SO; SO 2 or NR,; m is an integer from 0 to 3 inclusive; R. and R 6 each independently may be -F, -Br, -C0 2

R

3

-COR

3

-CON(R

3 2 -CN;

NO

2

-N(R

3 2

-OR

3 straight chained or branched alkyl, C-C 7 monofluoroalkyl, polyfluoroalkyl,

C

2 alkenyl,

C

2

-C,

alkynyl, C3-C, cycloalkyl, or C 3

-C

cycloalkenyl, wherein the alkyl, monofluoroalkyl, polyfluoroalkyl, alkenyl, alkynyl, cycloalkyl or cycloalkenyl may be unsubstituted or substituted -with aryl or heteroaryl; aryl or heteroaryl wherein the aryl or heteroaryl may be unsubstituted or substituted with -CI, -Br,

-NO

2 -CN, -N(R 3 2

-OR

3

-COR

3 -C0 2

R

3 or -CON(R 3 2 straight chained or branched CI-C, alkyl, monofluoroalkyl or polyfluoroalkyl, straight chained or branched C 2 alkenyl or alkynyl, C3_C, cycloalkyl, monofluorocycloalkyl, polyfluorocycloalkyl, or cycloalkenyl; R, may be straight chained or branched c 1 -C alkyl, hydroxyalkyl, aminoalkyl, alkoxyalkyl, monofluoroalkyl or SUB STITUTE S H EET (RULE 26) WO 97/17969 PCTIUS96/18573 polyf].uoroalkyl; straight chaiz'.ed or branched

C

2 alkenyl or alkynyl; C 3 cycloalkyl, monofluorocycloalkyl, polyfluorocycloalkyl or cycloalkenyl; -CN; -C0 2

,R

3 -CORI; c.r

O,

where R, is straight chained or branched

C

1 -c 1 alkyl, ronofluoroalkyl. or polyfluoroalkyl; straight chained or branched alkeny. or alkynyl; C 3

-C

7 cycloalkyl, mono fl1uorocyc loa lkyl, polyfluorocycloalkyl or cycloalkenyl; where R 9 may be -CN; straight chained or branched C 1 alkyl, monofluoroalky. or polyfluoroalkyl; straight chained or branched C 2

-C,

alkenyl or alkynyl; C 3 C- cycloalkyl, ronofluorocycloalkyl, polyfluorocycloalky. or cycloalkenyl; -N (RO) 2

-OR

3

(CH

2 PORI; CORI; -C0 2

R

3 or -CQN(R 3 2 wherein X may be S; 0; or NR 3 and wherein B may be straight chained or branched alkyl, mono fluoroalkyl, polyfluoroalkyl, alkoxy or thioalkyl; straight chained or branched

C

2 alkenyl; -SCH 2

C

6

H

4

OR

3

-(CH

2 6

H

5

-CM

2

X(CH

2 n1IR 3

(CH

2 ),N1{R 3 or -OR 3 or a pharmaceutically acceptable salt thereof.

Tn the present invention aryl includes benzyl, benzoyl, naphthyl, or phenyl. and heteroaryl includes pyridyl, thiophenyl, furanyl, pyrazinyl, pyrryl, naphthyl, in dolyl, imidazolyl, benzfurazanyl, benzfuranyl, quinolyl, aminophenyl, benzimidazolyl or 2-keto-lbenzimidazolinyl.

SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -11- The compounds of the present inventn may be present as enantiomers, disteriomers, isomers or two or more of the compounds may be present to form a racemic mixture.

Furthermore, the compounds of the present invention are preferably at least 80% pure, more preferably 90% pure, and most preferably 95% pure.

The invention further provides for the enantiomer of any of the compounds described herein which is a cis isomer or trans isomer. The invention also provides for the enantiomer of any of the compounds described herein which is a cis isomer or trans isomer.

In one embodiment of the invention the compound is not 3 ,4-Difluorophenyl)-1-(N-(2-(4-cyano-4-phenylcyclohexylamino)ethyl]}carboxamido-5-methoxycarbonyl-4methoxymethyl-2-oxo-1,2,3,6-tetrahydropyrimidine hydrochloride.

Ten-fold selectivity differences are a minimum, but one skilled in the art will appreciate that compounds can be found that collectively have almost infinitely variable selective profiles. Compounds collectively having all possible combinations of selectivities are intended within the scope of this invention, provided that each of these compounds has at least a ten-fold greater affinity for the a receptor over the and/or aD receptors.

Preferred embodiments of the present invention include a compound selected from the group consisting of: SUBSTITUTE SHEET (RULE 26) VO 97/17969 PCT/US96/18573 -12-

M

R1 R

R

2

R

3

M

x Xn q R 4

N

X

n

R

R

and

M

R R 4

N

R

2

B

In one preferred embodiment for the compounds described herein R, is R R

R

3 R and p is 2, 3 or 4.

In one preferred embodiment for the compounds described herein R, is SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -13- OR

R

1r I R C N L N 3 R 6 -7 The invention also provides for compounds having the following structure: M 0R

R

2 N O

R

H

In one preferred embodiment of the present invention, M is polyfluorophenyl and RP and RF are independently substituted or unsubstituted phenyl or pyridyl.

In a further embodiment of the present invention, R, is

-COCH

3

-COCH

3 or -CONH, R 2 is CH 2

OCH

3 methyl or ethyl; and R is H, -CN, or CO 2

CH,.

The invention provides for the preferred embodiment having the following structures:

F

O N N N 0

N

H

SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 9717969PCT/US96/18573 -14

H

~NN

N

NX&0

H

Il,0

H

N 0

H

SUBSTITUTE SHEET (RULE 26) WO 97/1 7969 PTU9/87 PCT/US96/18573 -Is-

A

in, SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 97/ 7969PCTIUS96/18573 -16- Hb~K

N

H

H

N 0

H

SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 97/ 7969PCT/US96/18573 -17-

H

N 0

H

0 ",0 and

H

The invention al.so provides for compounds having the following structure: S USSTITUTE SH EET (RU LE 26) WO 97/17969 PCT/US96/18573 -18 M 0.

RR

P,

R

In another embodiment of the present invention M is polyfluorophenyl and R. is substituted or unsubstituted phenyl or pyridyl.

The invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the compounds described above and a pharmaceutically acceptable carrier. In the subject invention a "therapeutically effective amount" is any amount of a compound which, when administered to a subject suffering from a disease against which the compounds are effective, causes reduction, remission, or regression of the disease. In one embodiment the therapeutically effective amount is an amount from about 0.01 mg per subject per day to about 500 mg per subject per day, preferably from about 0.1 mg per subject per day to about 60 mg per subjec-t per day and most preferably from about 1 mg per subject per day to about 20 mg per subject per day. in the practice of this invention the "pharmaceutically acceptable carrier" is any physiological carrier known to those of ordinary skill in the art useful in formulating pharmaceutical compositions.

In one preferred embodiment the pharmaceutical carrier may be a liquid and the pharmaceutical composition would be in the form of a solution. In another equally preferred embodiment, the pharmaceutically acceptable SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -19carrier is a solid and the composition is in the form of a powder or tablet. In a further embodiment, the pharmaceutical carrier is a gel and the composition is in the form of a suppository or cream.

The invention provides a method of treating a subject suffering from benign prostatic hyperplasia which comprises administering to the subject any one of the compounds described herein effective to treat benign prostatic hyperplasia. The invention further provides that the compound additionally does not cause a fall in blood pressure at dosages effective to alleviate benign prostatic hyperplasia. In one preferred embodiment the compound effects treatment of benign prostatic hyperplasia by relaxing lower urinary tract tissue and in particular where the lower urinary tract tissue is prostatic smooth muscle.

The invention further provides a method of treating a subject suffering from elevated intraocular pressure which comprises administering to the subject one of the compounds described herein effective to lower intraocular pressure.

The invention further provides a method of treating a subject suffering from a disorder associated with elevated blood cholesterol which comprises administering to the subject one of the compounds described herein effective to inhibit cholesterol synthesis.

The invention also provides a method of treating a disease which is susceptible to treatment by antagonism of the receptor which comprises administering to the subject one of the compounds described herein effective to treat the disease.

The invention further provides a method of treating a subject suffering from impotency which comprises SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 administering to the subject one of the compounds described herein effective to treat impotency.

The invention further provides a method of treating a subject suffering from sympathetically mediated pain which comprises administering to the subject one of the compounds described herein effective to treat sympathetically mediated pain.

The invention provides a method of treating a subject suffering from cardiac arrhythmia which comprises administering to the subject one of the compounds described herein effective to treat cardiac arrhythmia.

This invention also provides a method of treating a subject suffering from benign prostatic hyperplasia which comprises administering to the subject an amount of any of the compounds described above in combination with a 5 alpha-reductase inhibitor effective to treat benign prostatic hyperplasia. Preferably, the reductase inhibitor is finasteride. The dosage administered to the subject is about 0.01 mg per subject per day to 50 mg per subject per day of finasteride in combination with an a1 antagonist. A preferred dosage administered to the subject is about 0.2 mg per subject per day to 10 mg per subject per day of finasteride in combination with an a, antagonist.

A more preferred dosage administered to the subject is about 1 mg per subject per day to 7 mg per subject per day of finasteride in combination with an a1 antagonist. The most preferred dosage administered to the subject is about 5 mg per subject per day of finasteride in combination with an aA antagonist.

The invention also provides for a pharmaceutical composition comprising a therapeutically effective amount of a combination of any of the compounds described herein in combination with finasteride and a pharmaceutically acceptable carrier. In one embodiment SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/1 8573 -21the pharmaceutical composition is a therapeuticallv effective amount of a combination comprising an amount from about 0.01 mg per subject per day to about 500 mg per subject per day of any one of the compounds described herein and an amount cf finasteride of about mg per subject per day. A more preferred embodiment of the pharmaceutical composition is a therapeutically effective amount of a combination comprising an amount from about 0.1 mg per subject per day to about 60 mg per subject per day of any one of the compounds described herein and an amount of the finasteride of about 5 mg per subject per day. The most preferred embodiment of the pharmaceutical composition is a therapeutically effective amount of a combination comprising from about 1 mg per subject per day to about mg per subject per day of any one of the compounds described herein and an amount of the finasteride of about 5 mg per subject per day.

The invention further provides a method of relaxing lower urinary tract tissue which comprises contacting the lower urinary tract tissue with an amount of one of the compounds described herein effective to relax lower urinary tract tissue. In one embodiment the lower urinary tract tissue is prostatic smooth muscle. In one preferred embodiment the compound additionally does not cause a fall in blood pressure when it is effective to relax lower urinary tract tissue.

The invention provides a method of relaxing lower urinary tract tissue in a subject which comprises administering to the subject an amount of one of the compounds described herein effective to relax lower urinary tract tissue. In one preferred embodiment the compound does not cause a fall in blood pressure and the lower urinary tract tissue is prostatic smooth muscle.

The invention further provides for a method of SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -22inhibiting contraction of prostatic tissue, which comprises administering to the subject an amount of any of the compounds described herein effective to inhibit contraction of prostatic tissue. In one preferred embodiment the prostatic tissue is prostatic smooth muscle and the compound additionally does not cause a fall in blood pressure.

The invention provides for the use of the compounds described herein for the preparation of a pharmaceutical composition for lowering intraocular pressure, inhibiting cholesterol synthesis, and the treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the aQA receptor may be useful. The invention provides for the use of the compounds described herein for the preparation of a pharmaceutical composition for relaxing lower urinary tract tissue and in particular prostatic smooth muscle. The invention further provides for the use of any of compounds described herein for the preparation of a pharmaceutical composition, where the compound additionally does not cause a fall in blood pressure at dosages effective to lower intraocular pressure, to inhibit cholesterol synthesis, and for the treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the ar receptor may be useful.

The invention provides for the use of the compounds described herein in the preparation of a medicament for lowering intraocular pressure, inhibiting cholesterol synthesis, and for the treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the aA receptor may be useful. The invention provides for the use of the compounds described herein in the preparation of a medicament for relaxing lower urinary tract tissue and in particular prostatic smooth muscle. The invention SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -23further provides for the use of any of compounds described herein in the preparation of a medicament, where the compound additionally does not cause a fall in blood pressure at dosages effective to lower intraocular pressure, to inhibit cholesterol synthesis, and for the treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the a. receptor may be useful.

The invention provides for a drug which is useful for lowering intraocular pressure, inhibiting cholesterol synthesis, and the' treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the al receptor may be useful, the effective ingredient of the said drug being any of the compounds described herein. The invention further provides the drug described herein additionally does not cause a fall in blood pressure at dosages effective to lower intraocular pressure, to inhibit cholesterol synthesis, and for the treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the a, receptor may be useful.

The invention provides for a drug which is useful for relaxing lower urinary tract tissue and in particular prostatic smooth muscle, the effective ingredient of the drug being any of the compounds described herein.

The invention further provides the drug which is useful for relaxing lower urinary tract tissue additionally does not cause a fall in blood pressure at dosages effective to relax lower urinary tract tissue.

The invention also provides for the and enantiomers of all compounds of the subject application described herein. Included in this invention are pharmaceutically acceptable salts and complexes of all of the compounds described herein. The salts include but are not limited to the following acids and bases.

SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -24- The following inorganic acids; hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, sulfuric acid and boric acid. The organic acids; acetic acid, trifluoroacetic acid, formic acid, oxalic acid, malonic acid, succinic acid, fumaric acid.

tartaric acid, maleic acid, citric acid, methanesulfonic acid, trifluoromethanesulfonic acid, benzoic acid, glycolic acid, lactic acid and mandelic acid. The following inorganic bases; ammonia, hydroxyethylamine and hydrazine. The following organic bases; methylamine, ethylamine, propylamine, dimethylamine, diethylamine, trimethylamine, triethylamine, ethylenediamine, hydroxyethylamine, morpholine, piperazine and guanidine. This invention further provides for the hydrates and polymorphs of all of the compounds described herein.

In one preferred embodiment the pharmaceutical carrier may be a liquid and the pharmaceutical composition would be in the form of a solution. In another equally preferred embodiment, the pharmaceutically acceptable carrier is a solid and the composition is in the form of a powder or tablet. In a further embodiment, the pharmaceutical carrier is a gel and the composition is in the form of a suppository or cream. In a further embodiment the compound may be formulated as a part of a pharmaceutically acceptable transdermal patch.

A solid carrier can include one or more substances which may also act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aids, binders or tablet-disintegrating agents; it can also be an encapsulating material. In powders, the carrier is a finely divided solid which is in admixture with the finely divided active ingredient.

In tablets, the active ingredient is mixed with a carrier having the necessary compression properties in SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain up to 99k of the active ingredient. Suitable solid carriers include, for example, calcium phosphate, ~magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins.

Liquid carriers are used in preparing solutions, suspensions, emulsions, syrups, elixirs and pressurized compositions. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fats. The liquid carrier can contain other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmoregulators. Suitable examples of liquid carriers for oral and parenteral administration include water (partially containing additives as above, e.g.

cellulose derivatives, preferably sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g. glycols) and their derivatives, and oils fractionated coconut oil and arachis oil). For parenteral administration, the carrier can also be an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are useful in sterile liquid form compositions for parenteral administration. The liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellent.

Liquid pharmaceutical compositions which are sterile solutions or suspensions can be utilized by for example, intramuscular, intrathecal, epidural, SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -26in=raperitoneal or subcutaneous injection. Sterile solutions can also be administered intravenously. The compounds may be prepared as a sterile solid composition which may be dissolved or suspended at the time of administration using sterile water, saline, cr other appropriate sterile injectable medium. Carriers are intended to include necessary and inert binders, suspending agents, lubricants, flavorants, sweeteners, preservatives, dyes, and coatings.

The compound can be administered orally in the form of a sterile solution or suspension containing other solutes or suspending agents, for example, enough saline or glucose to make the solution isotonic, bile salts, acacia, gelatin, sorbitan monoleate, polysorbate (oleate esters of sorbitol and its anhydrides copolymerized with ethylene oxide) and the like.

The compound can also be administered orally either in liquid or solid composition form. Compositions suitable for oral administration include solid forms, such as pills, capsules, granules, tablets, and powders, and liquid forms, such as solutions, syrups, elixirs, and suspensions. Forms useful for parenteral administration include sterile solutions, emulsions, and suspensions.

Optimal dosages to be administered may be determined by those skilled in the art, and will vary with the particular compound in use, the strength of the preparation, the mode of administration, and the advancement of the disease condition. Additional factors depending on the particular subject being treated will result in a need to adjust dosages, including subject age, weight, gender, diet, and time of administration.

SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCTIUS96/18573 -27- This invention further provides for metabolites of the compounds of the present invention. The in vivo activities and mechanisms of action of numerous enzymes responsible for the generation of metabolites of pharmaceutical compounds are well-known in the art. For example, ethers may be modified to alcohols, esters may be modified by esterases, or amides may be modified by amidases and peptidases.

This invention further provides a prodrug of the compounds disclosed herein. Knowledge of metabolic activities allows the design of prodrug compounds which, when administered to a subject, such as a human, are expected to yield metabolites which include the compounds of the present invention. For example, the cyclohexylamino nitrogen may be modified by various substituents, such as methyl, alkanoyl, aroyl, or an alkyl or aryl carbamate may be formed, which are expected to yield the compounds of the present invention when acted upon in vivo by endogenous enzymes.

As stated above, such modifications are intended only as illustrative examples, and are not intended to limit the scope of the present invention, as such modifications and techniques therefore are well-known in the art.

SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -28- One skilled in the art will readily appreciate that appropriate biological assays will be used to determine the therapeutic potential of the claimed compounds for treating the above noted disorders.

This invention will be better understood from the Experimental Details which follow. However, one skilled in the art will readily appreciate that the specific methods and results discussed are merely illustrative of the invention as described more fully in the claims which follow thereafter.

Experimental Details For Examples 1-17 Scheme 1 illustrates the general synthetic scheme for the preparation of the compounds of the present invention. Specific references to synthetic steps from Schemes 2-8 are included in the Examples below where appropriate. All NMRs were obtained using a 300 MHz GE QEPLUS NMR machine.

Example 1 cis-6- (3,4-Difluorophenyl) [2-(4-cyano-4-phenylcyclohexylamino) ethyl] 4 -methyl-2-oxo-1,2,3,6-tetrahydropyrimidine hydrochloride.

Part-1 a) General procedure for 2-[4-Cyano-4-arylcyclohexylamino] ethylamine (Scheme 2; Part-1).

A mixture of 4-cyano-4-aryl-cyclohexanone (48.7 mmol) and ethylenediamine (8.78 g, 146 mmol) and ptoluenesulfonic acid (92 mg) in benzene (200 mL) was refluxed for 4 h in Dean-Stark trap to remove the water that formed. Solvent was evaporated and the residue was redissolved in methanol (60 mL) and cooled to 0 °C.

Sodium borohydride (6.4 5 g) was added in portions and SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -29the mixture was stirred at room temperature for 3 h.

Solvent was evaporated, the residue was dissolved in dichloromethane (300 mL), washed with brine (3 X 500 mL), dried (potassium carbonate), and the solvent evaporated to leave the product as a pale yellow viscous oil The product was found to contain the cis/trans isomers in the ratio of about 9:1.

Careful chromatography of this mixture with chloroform/methanol/2M ammonia in methanol (100/10/5 to 100/20/10) yielded several some earlier fractions enriched in the trans isomer with respect to the amino and cyano groups. Later fractions eluted contained almost pure cis isomer relative to the amino and cyano groups.

Part-2 b) Methyl 2-{(3,4-difluorophenyl)methylene}-3oxobutyrate (Scheme 2; Step A mixture of 3,4difluorobenzaldehyde (14.2 g, 0.1 mol), methyl acetoacetate (12.2 g, 0.105 mol), piperidine (0.430 g, mmol), and acetic acid (0.30 g, 5 mmol) in benzene (150 mL) was stirred and refluxed with a Dean-Stark trap for 8 hours. Benzene was evaporated, the residue was dissolved in ethyl acetate (200 mL) and washed with brine (50 mL), saturated potassium bisulfate solution mL), and saturated sodium bicarbonate solution in sequence. The ethyl acetate solution was dried (magnesium sulfate), solvent removed under reduced pressure and the residue was purified by column chromatography (Si02, EtOAc/hexane, 10-15%). The product, methyl 2-{(3,4-difluorophenyl)methylene}-3oxobutyrate, was obtained as a yellow oil (0.98 g, 98.3%) and was used in the next step without any further characterization.

c) 6-(3,4-Difluorophenyl)-1,6-dihydro-2-methoxy- 5-methoxycarbonyl-4-methylpyrimidine (Scheme 2; Step SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 A mixture of methyl 2 -((3,4-difluorophenyl) methylene}-3-oxobutyrate (8.8 g, 36.6 mmol), 0methylisourea hydrogen sulfate (9.4 g, 55 mmol), and NaHCO (12.3 g, 0.146 mol) in DMF (30 mL) was stirred and heated at 70 OC for 16 hours. The mixture was cooled, diluted with EtOAc (300 mL) and washed with water (5 X 300 mL), brine (300 mL), and dried (MgSO0 4 Solvent was evaporated and the crude product was purified by flash column chromatography on silica gel using 10% through 20% EtOAc in hexane as the gradient eluent, to leave the product as an oil (3.82 g, 30.2%).

d) 6-(3,4-Difluorophenyl)-1,6-dihydro-2-methoxy- 5-methoxycarbonyl-4-methyl-1-l [(4-nitrophenylozy) carbo nyl]pyrimidine (Scheme 2; E).

To a solution of 6-(3,4-difluorophenyl)-1,6-dihydro-2methoxy-5-methoxycarbonyl-4-methylpyrimidine (2.82 g, 9.52 mmol) and 4-dimethylaminopyridine (1.16 g, 9.52 mmol) in CH 2 C1, (50 mL),at 0-5 OC, 4-nitrophenyl chloroformate (1.82 g, 9.04 mmol) was added and the mixture was allowed to warm to room temperature. After 12 hours solvent was evaporated and the residue was purified by flash column chromatography (SiO,, EtOAc/hexane, 10%-15%)to obtain the product as white crystals (3.72, m.p. 172-174 OC.

a) 6-(3,4-Difluoropheny)-1,2,3,6-tetrahydro-2-oxo-5methoxycarbonyl-4-methyl-1-(4-nitrophenoxy) carbonylpyrimidine (Scheme 2; Step F).

To a well stirred solution of 6-(3,4-difluorophenyl)- 1,6-dihydro-2-methoxy-5-methoxycarbonyl-4-methyl--(4nitrophenoxy)carbonylpyrimidine (10 g) in THF (200 mL) at room temperature was added aqueous 6N hydrochloric acid (10 mL). The stirring was continued for 3 h.

Solvent was evaporated and the residue was dried under vacuum to obtain the product as a white powder (9.7 g, 100%); m.p. 185-186 oC.

SUBSTITUTE SHEET(RULE 26) WO 97/17969 PCTUS96/18573 -31- Part-3 f) cis-6-(3,4-Difuorophenyl)-).N-[2-(4-cy 4 p h e n y I cyclohexyl amino) ethyl] 4-methyl-2-oxo-1,2,3, 6 -tetrahydropyrimidine hydrochloride (Scheme 2; Step A solution of methoxycarbonyl-4-methyl-,2,3,6-tetrahyd 2 (3,4-difluorophenyl)-1-[(4-nitrophenyloxy)carbonyll pyrinidine (500 mg, 1.15 mmol) and cis-2-[4cyano- 4 -phenyl-cyclohexylamino]ethylamine (351 mg, mmol) in dichioromethane (50 mL) was stirred at room temperature for 12 hours. The solution was washed with ice-cold 0.SN NaOH (2 X 10 mL) and dried over sodium sulfate. Solvent was evaporated at reduced pressure and the residue was purified by flash chromatography on silica gel (dichloromethane:MeOH:2M ammonia in MeOH, 90:8:4) to give 0.55 g (871) as a white powder. The HCl salt was prepared by treatment of a solution of the free base in ether with IN HCl in ether. M.P. 172-174 Anal. Calcd. for C 29

H

3 jNS0 4

F

2 C1.1.3H 2 0:C1 56.96; H, 5.70; N, 11.45. Found: C, 57.10; H, 5.69; N, 11.12.

Example 2 cia-6-( 3 ,4,S-Trifluorophnyl)-l-{N-[2-(4-cyano-4phanyl-cyclhexylamino) ethyl])carboxamido-5..acetyl...4..mothoxymethyl 2 1,2,3,6-tetrahydropyrimidin. hydrochloride a) 3-((3,4,5-Trifluorophenyl)mtyl.ne.

2 4 pentanedione (Scheme 2; Step A mixture of 3,4,5trifluorobenzaldehyde (4.2 g, 26.2 mmol), 2,4pentanedione (2.62 g, 26.2 mmol), piperidine (0.430 g, mmol)in benzene (150 mL) was stirred and refluxed with a Dean-Stark trap for 8 hours. Benzene was evaporated and the yellow oily residue, trifluorophenyl) methylene I 4 -pentanedione, was used in the next step without any further purification.

SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -32b) 6-(3,4,5-Trifluorophenyl)-1,6-dihydro-2-methoxy- 5-acetyl-4-methylpyrimidine (Scheme 2; Step

A

mixture of 3 ,4,5-trifluorophenyl)methylene-2,4 pentanedione (26.2 mmol), O-methylisourea hydrogen sulfate (3.22 g, 39.3 mmol), and NaHCO, (6.6 g, 78.6 mmol) in EtOH (400 mL) was stirred and heated at 95-100 oC for 6 hours. The mixture was filtered and the solid residue was washed with ethanol (100 mL). Solvent was evaporated from the combined filtrate and the crude product was purified by flash column chromatography on silica gel using 10% through 25% EtOAc in hexane as the gradient eluent, to leave the product as an oil (2.80 g, 36%) c) 6-(3,4,5-Trifluorophenyl)-1,6-dihydro-2-methoxy- 5-acetyl-4-methyl-l-[ 4 -nitrophenyloxy)carbonyljpyrim idine (Scheme 2; Step E).

To a solution of 6-(3,4,5-trifluorophenyl)- 1,6-dihydro-2-methoxy-5-acetyl-4-methylpyrimidine (2.8 g, 9.38 mmol) and pyridine (10 mL) in CH 2 C1 2 (200 mL) at OC was added 4-nitrophenyl chloroformate (1.886 g, 9.38 mmol) and the mixture was allowed to warm to room temperature. After 12 hours solvent was evaporated and the residue was purified by flash column chromatography (Si02, dichloromethane/EtOAc, 10%-15%)to obtain the product as a white powder (4.0 g, 92%).

d) 6 3 ,4,5S-Trifluorophnyl)-1,2,3,6-ttrahydro-2-oxo- 5-acetyl-4-methyl-l- 4 -nitrophnyloxy)carbonyl]pyrim idine (Scheme 2; Step F).

To a well-stirred solution of 6-(3,4,5-trifluoro phenyl)-1,6-dihydro-2-methoxy-5-acetyl-4-methyl-l- [(4-nitrophenyloxy)carbonyl]pyrimidine (4.0 g, 8.63 mmol) in THF (100 mL) at 0-5 OC, 6N aqueous HC1 (4 mL) was added and the mixture was allowed to warm to room temperature. After 2 h, solvent was evaporated and the product dried under vacuum. The title compound was SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -33obtained as a pure single component and used un the next step without further purification (3.88 g, 100%).

e) ciB-6-(3,4,5-Trifluorophenyl)-1-{N-[2-(4-cyano-4.

phenyl-cyclohexylamino)ethyll)carboxamido-5-acetyl-4-methoxymethyl-2-oxo- 1,2,3,6-tetrahydropyrimidine hydrochloride (Scheme 2; Step G).

A mixture of 6-(3,4,5-trifluorophenyl)-1,2,3,6-tetra hydro-2-oxo-S-acetyl-4-methyl-l- (4-nitrophenyloxy) carbonyllpyrimidine (44.9 mg, 0.1 mmol) and cis-2-(4cyano-4-phenyl-cyclohexylamino) ethylamine (23.4 mg, 0.23 mmol) in THF (10 mL) was stirred at room temperature for 10 h and the solvent evaporated. The residue was redissolved in dichioromethane (10 nL), washed with ice-cold 0.5 N NaOH (2 X 5 mL), dried and solvent evaporated. The residue was purified by preparative thin layer chromatography on silica gel using chloroform-methanol-2M ammonia in methanol (100/2/1) as the eluent to afford the product as a white powder (53 mg, 881). The HCl salt was prepared by treatment with 1N HCl in ether to give the product as a white powder. Anal. Calcd. for C,,H,,N,OC1F, 2 C1 2 C, 56.66; H, 5.30; N, 10.83. Found: C, 56.51; H, 5.12; N, 11.09.

Example 3 (+)-cia-5-Carboxamido-6-(2,4-difluorophenyl)-1-{N- 2- cyan 0-4-ph. n y 1cyclohexylanino)ethyl])carboxamido-4-ethyl-2-oxo- 1,2,3,6-tetrahydropyrimidine hydrochloride.

a) Beuzy 2-[(2,4-difluorophenyl) methylen.I-3-oxopentanoate (Scheme 2; Step A solution of benzyl propionylacetate (157 g, 0.758 mol), 2,4-difluorobenzaldehyde (107.65 g, 0.758 mol), and piperidinium acetate (5.49 g, 38 mmol) in benzene (1 L) SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 9717969PCT/US96/18573 -34were stirred at room temperature for 96 h. The mixture was washed with water (2 X 100 rnL) dried (magnesium sulfate) and the solvent evaporated under reduced pressure to yield the product as a pale yellow syrup (251.2 g) which was used in the next step withcut further purification.

b) 5- (Benzyloxycarbonyl) 6-dihydro-2-methoxy-4-ethyl- 6- 4-difluorophenyl) pyrimidine (Scheme 2; Step D).

A suspension of benzyl 2-[(2,4-difluorophenyl) methylene]-3-oxopentanoate (80.0 g, 0.241 mol), 0methylisourea hemisulf ate (63.8 g, 0. 362 rnol, 1. 5 eq.), NaHCO 3 (60.48 g, 0.72 mol) in ethanol (800 rnL) was stirred at 60-700"C for 20 h. After cooling to room temperature, the mixture was filtered, and the solid was washed with ethanol (200 mL). The solvent was evaporated from the combined filtrates and the residue was purified by column chromatography (SiO 2 1 EtOAc/Hexane, 10%-30%) to yield 5- (benzyloxycarboiyl) l,6-dihydro-2-methoxy-4-ethyl-6- (2,4-difluorophenyl) pyrimidine as a pale yellow oil (39 g, 1H-NMR analysis showed the product to be a mixture of amine/imine tautomers, which was used as is in the next step.

c) 5- (Beziyloxycarbziyl) -4-ethyl-i, 6-dihydxro-2-mothomy- 6- 4-difluorophenyl)-1- [(4-nitrophenyloxy) carbonyllpyrizidine (Scheme 2; Step F) To a well stirred solution of 5- (benzyloxycarbonyl) 6-dihydro- 2 -methoxy-4 -ethyl-6 4-dif luorophenyl) pyrimidine (22.5 g, S9.3 mmol) and 4 N -dimethyl amino) pyridine (9.3 g, 75.8 mmol) in CH 2 Cl 2 (200 mL) was added powdered 4-nitrophenyl chioroformate (15.3 g, 75.8 mmol) at 0 0

C.

The reaction mixture was stirred for 12 h at room temperature and then water (50 mL) was added. The pH of the aqueous layer was adjusted to 10-11 by the addition of 6 N sodium hydroxide. The dichioromethane SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCTIUS96/18573 layer was separated and dried (Na 2

SO

4 Solvent was evaporated in vacuc and the residue was purified by column chromatography kSiO 2 dichioromethane/hexane, 20%-50%) to yield the product as a viscous oil (32.C g, 981).

d) 5- (Benzyloxycarbonyl) 4 -ethyl-1,6-dihydro 1 £2phenyl)ethyll)carboxamido-2-mothoxy-6-(2,4-difluoro phenyl)pyrimidine (Scheme 3; Step. C-1 To a stirred solution of 5-(benzyloxycarbonyl)-4-ethyl-, 6dihydro-2-methoxy-6-(2,4-difluorophenyl) 1 4 nitrophenyloxy) carbonyl]pyrimidine (32 g, 58.17 mmol) in dichloromethane (200 mL) was added methylbenzylamine (9.16, 75.6 mmol) at room temperature. The mixture was stirred for 12 h and then was diluted with more dichloromethane (200 mL) and washed with 0.5 N NaOH solution (2 x 60 mL). The organic layer was dried over Na 2 SO., filtered and solvent was evaporated. The resulting mixture of diastereomers was separated by column chromatography (SiO 2 3% EtOAc in toluene). The first major product to elute was (.)-5-(benzyloxycarbonyl)-4-ethyl-1,6dihydro-i-{N- (2-phenyl) ethyl] }carboxamido-2-methoxy-s- (2,4-difluorophenyl)pyrimidine (12.15 g, 38%) +214 (c 1.5 g in 100 mL CHC.

3 e) (+)-5-(Benzyloxycarbonyl)-1,6-dihydro-2-methoxy-4ethyl-6-(2,4-difJurophenyl) pyrimidine (Scheme 3; Step To a stirred solution of -5 (benzyloxycarbonyl) -4-ethyl-1,6-dihydro-1-{N- [2-phenyl)ethyl] }carboxamido- 2-methoxy-6-( 2 '4-difluorophenyl)pyrimidine (11.15 g, 20.41 mmol) in toluene (250 mL) was added 1,8diazabicyclo[5,4,o]-undec-7-ene (4.04 g, 26.53 mmol) and the mixture was stirred at room temperature for 14 h. The solvent was evaporated and the residue was purified by flash column chromatography on silica gel with 3:1 EtOAc/hexane as eluent to give SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -36- (benzyloxycarbonyl)-1,6-dihydro-2-rethoxy-4-ethyp6 (2,4-difluorophenyl)pyrimidine as a viscous oil (6.15 g, 78%).

f) (+)-S-(Benzyloxycarbonyl)-4-ethyl-1,6-dihydr 0 2 methoxy-6-(2,4-difluorophenyl)-1- (4-nitropheny1 -oxy) carbonyllpyrimidine (Scheme 2; Step To a well stirred solution of (+)-5-(benzyloxycarbonyl)-1,6dihydro-2-methoxy-4-ethyl-6-(2,4-difluorophenyl) pyrimidine (4.1 g, 10.62 mmol) and 4-(N,Ndimethylamino) pyridine (1.69 g, 13.80 mmol) in CH,Cl- (200 mL) was added solid 4-nitrophenyl chioroformate (2.78 g, 13.80 mmcl) at room temperature. The reaction mixture was stirred for 12 h and washed with 0.5 N NaOH solution (2 X 50 mL). The organic layer was separated and dried (Na 2 The solvent was evaporated and the residue was purified by column chromatography on silica gel using dichioromethane/hexane (20%-50t) as the eluent to give 5- (benzyloxycarbonyl) -4 -ethyl- 1, 6 dihydro-2-methoxy-6-(2,4-difluorophenyl)l[(4nitrophenyloxy)carbonyl pyrimidine (5.37 g, 92%) as a viscous oil.

g) -5-Benzyloxycarbonyl-4-ethyl-1,2,3,6tetahydro2 oo-6-(2,4-ditluorephenyl)-l-[(4-nitrophenyloxy) carbonyllpyrimidine (Scheme 4; Step A).

To a solution of (4)-6-(2,4-difluorophenyl)-1,6dihydro-2-methoxy-5-benzyloxycarbonyl-4ethy1- (4nitrophenoxy) -carbonylpyrimidine (6.50 g, 11.81 mmcl) 3C in dichloromethane (150 mL) at room temperature was added IN HCl in ether (50 mL) and the mixture was stirred at room temperature for 12 hours. Solvent was evaporated at reduced pressure and the residue was dried to give 6.31 g (100) of the product as a white powder.

h) (Benzyloxycarbonyl) (2,4-difluoophenyl) -4- SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -37ethyl-2-oxo-1,2,3,6-tetrahydro-1-{N- 12-(4-cyano-4phenyl-cyclohexylamino)ethyl] carboxamidopyrimidine (Scheme 4; Step B-1).

A mixture of (+)-5-benzyloxycarbonyl-4-ethyl-1,2,3,6- Sezrahydro-2-oxo-6- (2,4-difluorophenyl)-i nitrophenyloxy)carbonyllpyrimidine (537 mg, 1 mmol) and cis- 2 (4 cyano-4 -phenyl-cyclohexylamino) ethylamine (281 mg, 1.2 mmol) in THF (20 mL) was stirred at room temperature for 10 h and the solvent evaporated. It was redissolved in dichloromethane (10 mL), washed with ice-cold 0.5 N NaOH (2 X 5 mLt), dried and solvent evaporated. The residue was purified by preparative thin layer chromatography on silica gel using chloroform-methanol-2M ammonia in methanol (100/2/1) as the eluent to afford the product as a white powder (603 mg, 94%) i)(+)-6-(2,4-Difluorophonyl)-4-ethyl-2-oxo-1,2,3,6tetrahydro-l-{N-(2-(4-cyano-4-phenylcyclohexylamino) ethyl] carboxylic acid (Scheme 4; Step B-2).

To a suspension of 10% Pd-C (100 mg) in MeOH (40 mL) and H 2 0 (8 mL) was added a solution of (benzyloxycarbonyl)-6-(2,4-difluorophenyl)-4-ethyl-2oxo-1,2,3,6-tetrahydro-1-{N-[2-(4-cyano-4-phenylcyclohexylamino)-ethyl] carboxamidopyrimidine (320 mg, mmol) in methanol (10 mL) and the mixture was hydrogenated at 80 psi for 6 h. The black suspension was filtered through a pad of celite and washed thoroughly with MeOH (2.0 L) and methanol/chloroform 200 mL). Solvent was evaporated from the combined filtrate to yield the product difluorophenyl)-4-ethyl-2-oxo-1,2,3,6-tetrahydro- 1- N- 2 4 c yano 4 pheny cyclohexylamino)-ethyll}carbox acid as a white solid (276 g, The product was used in the next step without SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -38further purification.

j cis- 5 Carboxamido 6- (2 4 difluorophenyl)-1-{N- 2-(4-cyano-4-phenylcyclohexylamino)ethyl]}carboxamido-4-ethyl-2-oxo- 1,2,3,6-tetrahydropyrimidine hydrochloride (Scheme 4; Step B-3).

A mixture of (+)-cis-6-(2,4-difluorophenyl)-4-ethyl-2oxo-1,2,3,6-tetrahydro-l-{N-[2-(4-cyano-4-phenylcarboxylic acid (141 mg, 0.25 mmol), 1-(3dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (71.3 mg, 0.372 mmol), and 4-(N,N-dimethylamino) pyridine (45.4 mg, 0.372 mmol) in anhydrous dichloromethane (30 mL) was stirred at room temperature for 2 h. 40% Aqueous ammonia (4090, 0.5 mL) was added to this mixture and the stirring was continued for 12 h. The mixture was diluted with 20 mL of dichloromethane and washed with saturated aqueous ammonium chloride solution (3 X 10 mL). Solvent was evaporated from the dried (sodium sulfate) dichloromethane solution and the residue was purified by column chromatography on silica gel using chloroform-methanol-2M ammonia in methanol (100/2/1) as the eluent, to obtain the desired product as a white powder (120 mg, [a]0 +107 (c 1.38 g, dichloromethane). The HC1 salt was prepared by treatment of a solution of the free base in ether with IN HC1 in ether. M.p. 225-228 OC; Anal. Calcd. for

C

2 9H 33

N

6 0 3

F

2 C1.0.38H 2 0.0.19CH 2 C1 2 57.47; H, 5.64; N, 13.75. Found: C, 57.80; H, 5.56; N, 13.37.

Example 4 (+)-cis-6-(3,4-Difluorophenyl)-l-{N-[2-(4-cyano-4p h e n y 1 4-methoxymethyl-2-oxo-1,2,3,6-tetrahydropyrimidine SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -39hydrochloride Part-2 a) Methyl 3 ,4-difluorophenyl)methylene]-3-oxo-4methoxybutyrate (Scheme 2; C).

To a solution of methyl 4 -methoxyacetoacetate (84.32 g, 0.577 mol), 3 4 -difluorobenzaldehyde (82 g, 0.577 mmol), and piperidinium acetate (5.86 g, 0.068 mol) in benzene (1.5 L) were added molecular sieves (400 g) and the mixture was stirred at room temperature for 48 h.

The molecular sieves were removed by filtration and the solvent was evaporated from the filtrate under reduced pressure. The residue was purified by column chromatography on silica gel using chloroform/ethyl acetate (100:3) to get the product as an oil (67 g, This product was a mixture of cis and trans isomers.

b) 5-Methoxycarbonyl-l,6-dihydro-2-methoxy-4methoxymethyl-6-(3, 4 -difluorophenyl)pyrimidine (Scheme 2; Step A suspension of methyl difluorophenyl)methylene]-3-oxo-4-methoxybutyrate (7.50 g, 27.75 mmol), O-methylisourea hemisulfate (7.17 g, 41.63 mmol, 1.5 and sodium bicarbonate (6.99 g, 83.25 mmol, 3 eq.) in ethanol (400 mL) was stirred at 50-55 OC for 6 h. The solvent was evaporated from the combined filtrates and the residue was purified by column chromatography (SiO 2 EtOAc/hexane, 10%-30%) to give 5-methoxycarbonyl-1,6-dihydro-2-methoxy-4methoxymethyl-6-(3,4-difluorophenyl)pyrimidine as a pale yellow oil (4.3 g, 47%).

c) 5-Methoxycarbonyl-4-methoxymethyl-1,6-dihydro-2methoxy-6-(3,4-difluorophenyl) 1 4 nitrophenyloxy)carbonyl]pyrimidine (Scheme 3; Step B).

To a well stirred solution of 5-methoxycarbonyl-1,6dihydro-2-methoxy-4-methoxymethyl-6-(3,4-difluoro SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCTIUS96/18573 phenyl)pyrimidine (4.3 g, 13.18 mmol) and 4-(N,Ndimethylamino) pyridine (2.09 g, 17.13 nmol) in CH 2 C1, (100 nL) was added solid 4-nitrophenyl chioroformate (3.45 g, 17.13 mmol) at 0 OC. The reaction mixture was stirred for 12 h at room temperature and the solid formed was removed by filtration. Solvent was evaporated from the filtrate and the residue was purified by column chromatography (SiO 2 dichloromethane/hexane, 20%-50%) to yield the product as a viscous oil (3.85 g, 59%).

d) 5-Methoxycarbonyl-4-methoxymethyl-1,6 -dihydro-l-{N- 2 -phenyl)ethyl]1carboxaido-2-methoxy-6 -(3 4 difluorophenyl)pyrimidine (Scheme 3; Steps C-1 C-2).

To a stirred solution of 5-methoxycarbonyl-4methoxymethyl-1,6-dihydro-2-methoxy-6-( 3 4 difluorophenyl)-l-(4-nitrophenyloxy)carbonyl) pyrimidine (3.82 g, 7.77 mmol) in THF (140 mL) was added R- -a-methylbenzylamine (1.13 g, 9.33 mmol, 1.2 eq.) at room temperature and the stirring was continued for 12 h. Solvent was evaporated and the residue was purified by column chromatography (SiO 2 10-201; EtOAc in hexane). The first major product to elute was methoxycarbonyl-4-methoxmethyl-, 6-dihydro-l- [2phenyl)ethyl])carboxamido-2 methoxy6 (3 4..

difluorophenyl)pyrimidine (1.74 g, 44.5t), as an oil.

WD +205.5 (c 5.1 g in 100 mL CHC1.).

e) (+)-5-Methoxycarbonyl-1,6-dihydro-2-methoxy-4methoxymet hyi-6 (3 ,4-difluorophenyl) pyrimidine (Scheme 3; Step C-3).

To a stirred solution of (+)-S-methoxycarbonyl-4methoxymethyl- 1,6-dihydro-l-(N- [2-phenyl)ethy.

carboxamido-2-methoxy-6-(3,4-difluorophenyl)pyrimidine (1.74 g, 3.67 mmol) in toluene (40 mL) was added 1,8diazabicyclo[5,4,01-undec-7-ene (0.250 g, 1.64 mmcl) and the mixture was stirred at 70-80 0 C for 1.5 h.

SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 97/ 7969PCTIUS96/18573 -41- Solvent was evaporated and the residue was purified by flash column chromatography on silica gel with 9: 1 CHC1 3 /EtQAc as eluent to give -S5-methoxycarbonyl 6 diflydro- 2 -methoxy-4.methoxmethyl.E 3 ,4-difluorc pheny'L)oyrimidine as a viscous oil (1.11 g,92.50s).

f) Chiral HPLC separation of 1, 6 -dihydro-2-mthoxy.4.methoxymethl..E,4 difluorophenyl)pyrimidine (Scheme 3; Step A).

The racemic S-methoxycarbonyl.., G-dihydro2-methoxy.

4 methoxymethyl-s- 4 -difluorophenyl) pyrirnidine was resolved by chiral HPLC [Chiralcel OD 20 X 250 mm #369- 703-30604; X 254 nm; hexanes/ethanol 95/5 and 0.01% diethylamine; 60 mg per injection; retention time of the desired enantiomer: 10.80 min., the first enantiomer peak to elutel to give methoxycarbonyl-1, G-dihydro-2-methoxy4-.methoxyrethyl- 6- (3,4 -difluorophenyl) pyrimidine.

g) (+)-S-Metho~carbony4methoxCymehyl..l 6-dihydro-2methoxy-6- 4 -difluorophenyl) 4 -nitrophenyl oxy)carbonyllpyriidine (Scheme 2; Step E) To a well stirred solution of lDE-dihydro-2-methoxy.4.methoxymethyl...G( 3 4 difluorophenyl)pyrimidine (1.11 g, 3.4 mmol) and 4- (N,N-dimethylamino)pyridine (0.54 g, 4.42 mmol) in

CH

2 Cl 2 (200 mL) was added powdered 4-nitrophenyl chioroformate (0.891 g, 4.42 mmcl) at room temperature.

Solvent was evaporated and the residue was purified by column chromatography on silica gel using CHCl,/EtOAc (20k-50%) as the eluent to give 4 -methoxymethyl-.1,6.dihydro.2methoxy6( 3 4 dif lurophenyl) 1- (4 -nitrophenyloxy) carbonyl) pyrimidine (1.30 g, 78k) as a viscous oil, (aID +262.2 (c 2.3 g in 100 ML CiCl 3 h) (+)-SMtoyaboy--ehxmehl1236 SUBSTITUTE S HEET (RULE 26) WO 97/17969 PCTIUS96/18573 -42tetrahydro-2-oxo.6.(3,.4difluoroph yl)l-[( 4 nitrophenyloxy)carbonyl]pyrimidine (Scheme 2; Step F).

To a Solution of (+)-6-(3,4-difluorophenyl) 1,6dihydro-2 -methoxy-5 -methoxycarbonyl-4 -methoxymethyl-l- (4-nitrophenoxy)-carbonylpyrimidine (0.305 g, 0.60 mmol) in dichioromethane (15 mL), iN HCl in ether (6 mL) was added at room temperature and the mixture was stirred at room temperature for .2 hours. Solvent was evaporated at reduced pressure and the residue was dried to give 0.295 g (100%) of the product as a white powder.

Part-3 i) 3 4 -Difluorophenyl) 4 -cyano-4p h e n y 1 cyclohexylamino) ethyl] 4-methoxymethyl-2-oxo-1,2,3,6-tetrahydropyz±midine hydrochloride (Scheme 2; Step A solution of methoxycarbonyl-4-methoxymethyl1l, 2 ,3,6-tetrahydro-2oxo-6-(3,4-difluorophenyl)- [(4-nitrophenyloxy) carbonyl]pyrimidine (0.850 g, 1.73 mmol) and cis-2-(4cyano-4-phenyl-cyclohecylamino) ethylamine (0.567 g, 2.42 mmol) in dichloromethane (50 mL) was stirred at room temperature for 12 hours. It was washed with icecold 0.SN NaOH (2 X 10 mL) and dried over sodium sulfate. Solvent was evaporated at reduced pressure and the residue was purified by flash chromatography on silica gel (dichloromethane:MeOH:2M ammonia in MeOH, 90:8:4) to give 0.905 g as a white powder. The HCl salt was prepared by treatment of a solution of the free base in ether with 1N HC1 in ether. The white powder was dried and recrystallized from anhydrous isopropanol. M.P. 233-235 0 C; [a]t +131.16 (c 0.735, MeOH); Anal. Calcd. for C,,H 34 N,OsF 2 Cl: C, 58.30; H, 5.53; N, 11.06. Found: C, 57.91; H, 5.53; N, 11.10.

Example SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 _43- (+)-trans-6-(3,4-Difluorophenyl)-l

(N

2 4 P h e n y 1 cyclohexylamino) ethyl] 4-methoxymethyl-2-oxo-1,2,3, 6 -tetrahydropyrimidine hydrochloride (Scheme 2; Step F).

A solution of (+)-S-methoxycarbonyl-4 -methxymethyl- 1,2,3,6-tetrahydro-2-oxo-6-(3,4-difluorophenyl)l[( 4 nitrophenyloxy)carbonylIpyrim dine (0.850 g, 1.73 mmol) and trans-2-( 4 -cyano-4-phenyl-cyclchexylamino) ethylamine (0.567 g, 2.42 mmcl) in dichioromethane

(SO

mL) was stirred at room temperature for 12 hours. it was washed with ice-cold 0.5N NaCH (2 X 10 mL) and dried over sodium sulfate. Solvent was evaporated at reduced pressure and the residue was purified by flash chromatography on silica gel (dichioromethane:MeOH:2M ammonia in MeOH, 90:8:4) to give 0.905 g (901) as a white powder. The HCl salt was prepared by treatment of a solution of the free base in ether with lN HCl in ether. Anal. Calcd. for C 3 H1 34

NO,F

2 C1:C 58.30; H, 5.53; N, 11.06. Found: C, 57.99; H, 5.73; N, 11.20.

Example 6 -cis-6- 4-Difluorphmyy) £2-(4-cyano-4p h e n 1cyclohexylamino) ethyl] )carboxamio-5-methoxycarbonyl 4-3sthoxymtthyl-2-oxo-l,2,3, 6 -tetrahydropyrimidine hydrochloride.

a) 5-Xethoxycarboyl-4 -methoxyethyl-1,6 -dihydro-2methoxy-6-( 3 4 -difluorophenyl)-l-((4nitrophanyloxy) carbonyllpyrimidine (Scheme 2; Step E).

To a well stirred solution of (+)-S-methoxycarbonyl- 1,6-dihydro-2-methoxy-4-methoxymethyl-6-( 3 4 difluorophenyl)pyrimidine (1.167 g, 3.576 mmol) and 4- (N,N-dimethylamino)pyridine (0.655 g, 5.364 mmol) in

CH

2 C1 2 (20 mL) was added powdered 4-nitrophenyl chioroformate (0.937 g, 4.65 mmol) at room temperature.

SUBSTITUTE SHEET (RULE 26) WO 97/1 7969 PCT/US96/18573 -44- Solvent was evaporated and the residue was Purified by column chromatography on silica gel using hexanes/EtCc as *'he eluent to give -5-methoxycarony..4 rethoxetylethy116dydro2mtoxy- 6 -34 diflurophenyl) 4 -nitrcrhenivloxy carbonyll pyrPi~idine (1.761 g, 100t) as a viscous oil. -335.3 (c g in 100 mL CHCd 3 b)(-)ci.6..(3,4-Difluorophey)lN[ 2 4 -cao4 phenylcycl1ohexyl amino) ethyl] }carboxamido- bo nyl1 4- me t ho xy me t hyl1 2- ox o 1,2,3, 6 -tetrahydropyrimidine hydrochloride (Scheme 2; G Step A solution of (-)-5-methoxycarbonyl.

4 methoxymethyl-.1,6dihydro2methoxy6( 3 4 diflurophenyl) -1-((4-nitrophenyloxy) carbonyllpyrimidine (0.103 g, 0.21 mmol) and cis-2-(4cyano- 4 -phenyl-cyclohexylamino) ethylamine (0.059 gT, 0.251 mmol) in dichioromethane (50 mL) was stirred at room temperature for 12 hours. It was washed with icecold 0. SN NaOH (2 X 10 mLs) and dried over sodium sulfate. Solvent was evaporated at reduced pressure and the residue was purified by flash chromatography on silica gel (dichloromethane:MeOH:2M ammonia in MeOH, 90:8.4) to give 0.121 g (100%) as an oil. It was dissolved in THF (2 m14, treated with 0.2 mL of 6N HC' and stirred at room temperature for 4 h. Solvent was evaporated and the residue was dried in vacua to give the product as a white powder (130 mg, l00t) M.P. 220- 222 0 C; (a] 0 -124.29 (c 0.535, MeOH) Anal. Calcd.

for C,,H 3 ,NsOsF 2 Cl: C, 58.30; H, 5.53; N, 11.06. Found: C, 57.20; H, 5.57; N, 11.30.

Example 7 -trans-6- 4.Dfuoropheny 1){N 2 4 2 pyridyl) -cyclohexylamnino) ethyl] methoxycarbonyl -4 -methoxymethyl.-2 -oxo 1,2,3, 6 -tetrahydzepyrimidine hydrochloride SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 97/ 7969PCTIUS96/18573 a) General procedure for 2 -arylcyclohexylaminol ethylamine (Scheme 2; Step A).

A mixture of 4-arylcyclohexancne~ (18.7 mmol) 4- 2 -pyr idyl) cvc lohexanone) and ethylenadiamine 149- minol) and p-tcluenesulfcnic acid (92 mg) i n benzene (200 m'L) was refluxed for 4 17 in :ean-Stark t-rap to remove the water that formed. Solvent was evaporated and the residue was redissolved in rezhanol -IL) an-d cooled t:o 0 00. To this, sodiurr borchydride (6.4 5 g) was added in prtions and ?-he mixture was stirr-ed ac room temperature for 3 h. Solvent waS evaorated, the residue was dissolved in dichloromethane (300 tt) washed with brine (30 500 rLl dried (potassium carbonate), and the solvent evaporated to leave the product as a pale yellow viscous oil (90-95t) The product was pjure and found to contain the cis/trans isomers. A caretful chromatography of this mixture with chloroform /methanol/ 2M amrmonia in methanol (100/10/5 to 100/20/10 gave so-me earlier fractions enriched in tha cis isomer. The fracticns eluted at the end were almost pure trans isomer.

b) ()C-m--34Dfurpoy)l{-2(-2 p y r i d y 1 cyc lehexyl amino) ethyl] )carboxazido- 5-methoxycarbonyl 4-methoymethyl.2-oxol, 2,3, 6-t-etrahydro pyrizidine hydrochloride (Scheme 2; Step G).

A solution of (*)-5-methoxycarbonyl4-.methoxymethyl.

3C l, 2 3 ,E-tetrahydro2oxo6(34difluorcphenyl)-lr( 4 n4-rophenyloxy)carbnylp.ind 91g, 0.1ml and crans- 2- (4 2 -pyridy2.cyclohexylamino) ethylamine mg, 0.136 mmol) in THF (10 tnt) was stirred at room temperature for 12 hours. It was washed with ice-cold C.SN NaCH (2 X 5 mL.) and dried over sodium sulfate.

Solvent was evaporated at reduced pressure and the residue was purified by preparative tlc on silica gel SUBSITUTE SHEET (RULE 26) WO 97/1 7969 PCTIUS96/18573 -46iethyl acetate:MeOH, 10:1) to gi.ve 43 mg (7Wk as a white powder. The FIC1 salt was prepared by treatment of a solution of the free base in ether with IN HC1 in ether. 180-184 11C; +186 (c 0.65, methanol); knal. Calcd. for C 2

UH

3 5N.O.F2C1.0.4CH,C1 2 51.33; H, 5.43; N, 9.54. Found: C, 51.31; H, 5.43; N,9.69.

Examples 8 and 9 cis-6- (3,4-Difluorophany~l) 2 (4-phenylcycl1ohexyl amino) ethyl] )carboxamido-S-mthoxycarbnyl 4-methyl-2 -oxo-l. 2.3, 6 -tetrahydropyrimidine hydrochloride and transa- 6 3,4- D ifluo rophenyl 1 NC 2 4 phenylcyclohexylamino) ethyl] )carboxamido- mohzcronl4mty 2oo1236-tetrahydro pyrimidine hydrochloride a) 2-r4-Phenylcyclohexyli-4rnolethylamine (Scheme Step A).

A mixture of 4-phenylcyclohexanone (4.00 g, 22.96 mmol) and ethylenediamine (1.66 g, 27.5 rnmol) and ptoluenesulfonic acid (437 mug) in benzene (200 mU) was ref luxed for 4 hi in Dean-Stark trap to remove the water that formed. Tic analysis indicated the completion of the reaction. Solvent was evaporated and the product was used in the next step without any further purification.

b) ci5..

6 3,4-Difluoropheuyl)..1.{N2.(4.

phenylcycl1ohexyl amino) ethyl] boy--~hl2ool2 36ttayrprmdn hydrochloride and trans (3 4-Dif lucrophonyl) 12 phanylcyclohexylanino) ethyl] )carkboxamido- carbonyl-4 -methyl-2-ozo-1, 2,3, 6-tatrahydropyrimidine SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCTIUS96/18573 -47hydrochloride (Scheme 5; Step A solutio z, methoxycarbonyl-4-methyl-1,2,3,6-etrahyd 3 4 -difluorophenyl)-1-f(4-nitrophenyloxy)carbonylj pyrimidine (120 mg, 0.269 mmol) and 2 -[4-phenylcyclohexyliminolethylamin (69.3 mg, 0.33 mmmcj) in dichioromethane (20 mL) was stirred at room temperature for 12 hours. Solvent was evaporated, the residue was redissolved in ethanol (10 mL) and cooled to 0 0 C. To this sodium borohydride (61 mg, 1.614 mmol) was added and the stirring continued for 2 h. Solvent was evaporated and the residue was purified by preparative tlc on silica gel using (dichloromethane:MeOH: 2

M

ammonia in MeOH, 100:8:4) as eluent. Two products were isolated, the upper band corresponds to cis-6-(3,4- Difluorophenyl)-l-{N-f2-(4 -phenyl-cyclohexylamino) ethyl] }carboxyamido5-metoxycarbonyl-4-methyl-2-oxo- 1 2 3 E-tetrahydropyrimidine (16 mg), and the lower -band to trans-6-(3,4-Diffluorophenyl)-llN-f2-( 4 phenylcyclohexylamino) ethyl] )carboxamido-S..methoxycar bonyL-4-methyl-2-oxol-el 3, -tetrahydropyrimidirie (42 mg). The HC1 salts were prepared by treatment of a solution of the free base in ether with IN HC1 in ether.

Example 8: The cis isomer: M.P 145-146 OC; Anal. Calod.

for C,, 3

H

33

N,

4 0F 2 Cl 0.8H 2 0:C, 58.24; H, 6.04; N, 9.70.

Found: C, 581.3; H, 5.83; N, 9.50.

Example 9: The trans isomer: M.P 156-157 OC; Anal.

Calcd. for C,H 3 1 3 N0 4

F

2 C 0.4CHC1 3 55.85; H, 5.51;

N,

9.17. Found: C, 55.57; H, 5.76; N, 9.10.

Example trans-6-(3 4-Difluorophenyl) 2 -(4 Phenylcyclohex l )ath y 1amno] ethyl] )carboxzamido-S-mkthoxycarbonyl -4-mthyl-2oxo-1,2,3,6- tetrahydzopyrimidin. hydrochloride (Scheme SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -48- Step

C).

A mixture of trans-6-(3,4-difluorophenvl)--{N-[2-(4p h e rn Y 1 cyclohexylamino)ethyl] }carboxamido-5-methoxycarbo 4-methyl-2-oxo-1,2,3,6-tetrahydropyrimidine -0 yrmdn (60 rr,, 0.114 mmol), methyl iodide (19.4 mg, 0.137 mmol, 1.2 and potassium carbonate (31.5 mg, 0.23 mmol) iracetone (10 mL) was stirred and refluxed for 12 h and the solid was removed by filtration. Solvent was evaporated from the filtrate and the residue was purified by flash chromatography on silica ael (dichloromethane:MeOH: 2 M ammonia in MeOH, 90:8:4) to give the product as an oil. The HC1 salt was prepared by treatment of a solution of the free base in. ether with 1N HCl in ether (14mg). M.P. 140-142 Anal.

Calcd. for C29,H3sN.O,F,C10.O33hexanes.0.59CHC13:C, 56.07; H, 5.99; N, 8.28. Found: C, 56.37; H, 6.21; N, 7.90.

Example 11 4-Difluorphanyl)-1-{N- 2- 4 me thoxycarbony 1 4 pheny cyclohexylamino)ethyllcarbox amido-5-mthoxycarbonyl-4-methoxymethyl-2-oxo- 1,2,3,6-tetrahydropyrimidine hydrochloride a) 2- 4 -Methoxycarbonyl -4 -phenylcYclohexylaminolethylamine (Scheme 2; Step

B).

A mixture of 2 4 -cyano 4 phenylcyclohexylaminoethylamine (2.34 9, 10 mmol) and concentrated sulfuric acid (20 mL) was heated at 80-85 OC for 10 h. It was cooled to room temperature, mixed with anhydrous methanol (200 mL), and refluxed for h. Solvent was evaporated and the residue was poured onto ice (200 g) and basified to pH 11 by addition 6N NaOH. It was extracted with dichloromethane (4 X 125 mL), dried (potassium carbonate) and solvent evaporated to leave the product as an oil (2.1 g, This SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -49- Pr duc' was a pure mixture of CIS and tzans -somers.

It was used in the next step without any further purification.

b) (+)-cis-6-(3,4Dif1 12- (4 -=Othoxycarbonyl -4-phenyl -c~clohexylamlin) etyll carboxamido-5-methoxycarbnyl-4-mthoxymethyl-2-oxo- 1,2,3# 6 -tetrahydropyrimdine hydrochloride (Scheme 2; Step G).

A solution cf Smethoxycarbonyl- 4 methoxyme-hyl- 1,2, 3 ,6-tetrahydro-2-Oxo-6-( 3 ,4-difluoropheny?)-1_I(4nitrophenyloxy)carbonyl pyrimidine (128 mg, 0.26 mmol) and 2- (4-methoxcarbonyl- 4 -phenylcyclohexylamino) ethylamine (70.mg, 0.26 mmol) in dichioromethane mL) was stirred at room temperature for 12 hours. It was washed with ice-cold 0.SN NaOH (2 X 5 mt) and dried over sodium sulfate. Solvent was evaporated at reduced pressure and the residue was purified by flash chromatography on silica gel (dichloromethane:MeOH: 2

M

ammonia in MeeO, 90:8:4) to give 78 mg (78t) as a white powder. The HCl salt was prepared by treatment of a solution of the free base in ether with 1N HC1 in ether. 112-114 OC; lItD +136 (c 0.S5, CHCl 3 Anal.

Calcd. for

C

3 jH 37

N

4

F

2 l1O-4CHlCL2 55.05; H, 5.56;

N,

8.18. Found: c, 55.05; H, 5.50; N, 8.16.

Example 12 3 ,4-Diflurophanyl)-l-{(.(2- 4 i4-diphnyl cyc ohexylamiu) e thylI )carboxamido- S -me thoxycarbonyl- 4 -mstoxymethyl..2 10-1,2,3,6trahydro pyrimidine hydrochloride a) 4 -DiPheUylcyclohaxylair 0 jo ethylamine (Scheme 2; Step

A).

A mixture of 4 ,4-diphenylcyclohexanone (2.5 g) and ethylenediamine (10 g) and P-toluenesulfonic acid (100 mg) in benzene (200 ML) was refluxed for 4 h in Dean- SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 Stark trap to remove the water that formed. Solvent was evaporated and the residue was redissolved in methanol (60 mL) and cooled to 0 0 C. TO this, sodium borohydride (2 g) asded To this, sodium brohydride (2 g) was added in portions and the Mlxiure was stirred at room temperature for 3 h. Solvent wa evaporated, the residue was dissolved i dichloromethane (300 mL), washed with brine (3 X 500 mL), dried (potassium carbonate) and the solvent evaporated to leave the product as a colorless viscous oil (2.94 g, 10O%) The 'H-NMR showed this product to De pure and was used in the next step without any further purification.

b) (+)-6-(3,4-Difluorophenyl)-1-{N 2- (4,4 diphenycyclohexylamino) ethyl] arbonyl-4-methoymethyl-2-oxo-1,2,3,6-tetrahydro pyriidine hydrochloride (Scheme 2; Step

G).

A solution of (+)-5-methoxycarbonyl-4-methoxymethyl- 1,2,3,6-tetrahydro-2-oxo-6-( 3 ,4-difluorophenyl)-1-[(4nitrophenyloxy)carbonyllpyrimidine (125 mg, 0.254 mmol) and 2- 4 ,4-diphenylcyclohexylamino)ethylamine (125 mg, 0.446 mmol) in THF (15 mL) was stirred at room temperature for 12 hours. It was washed with ice-cold NaOH (2 X 5 mL) and dried over sodium sulfate.

Solvent was evaporated at reduced pressure and the residue was purified by flash chromatography on silica gel (dichloromethane:MeOH: 2 M ammonia in MeOH, 90:8:4) to give 147 mg (92t) of the product as a white powder.

The HC1 salt was prepared by treatment of a solution of the free base in ether with IN HC1 in ether. 160-162

OC;

Ca), =+184 (c 0.68, methanol); Anal. Calcd. for C3sH 3

,N

4 OsF 2 C1.0.19CHIC12 61.68; H, 5.79; N, 8.18.

Found: C, 62.82; H, 5.87; N, 8.37.

Example 13 (+)-cis-6-(3,4-Difluorophenyl) -1-{yl N- 2-(4-cyano-4-(2- SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -51methoxycarbonyl-4-methyl-2-oxo-1,2,3,6tetrahydropyrimidine hydrochloride (Scheme 2; Step G).

A mixture of (+)-6-(3,4-difluorophenyl)-1,6-dihydro-5methoxycarbonyi-4-methyl-1-(4-nitro)phenoxycarbonyl-2- S pyrimidone (60 mg, 0.138 mmol), cis-2- [4-cyano-4-(2fluorophenyl)cyclohexylamino]ethylamine (50 mg, 0.19 mmcl) and methylene chloride (4 mL) was stirred at room temperature overnight. After being washed with iN NaH solution, the organic layer was concentrated and purified by preparative TLC (eluent: 10/1 ethyl acetate/2M ammonia in methanol) to give the title compound in 41t yield (32 mg) as a yellow foam: DCIMS, m/z 570 (MW) Treatment of the free base with 1 equivalent of IM HC1 in ether gave the HC1 salt as an off-white solid: m.p. 172-176 0 C. 114.7 (1.9 mg/mL MeOH). Anal. Calc. for C 2 ,H,,FNsO, HC 0 SCHCl 3

C,

53.22; H, 4.77; N, 10.52. Found: C, 53.34; H, 4.77; N, 10.45.

Example 14 6 -(3,4-Difluorophenyl)-l-{N-12-(4-cyano-4-(2methoxycarbonyl-4-methoxyethyl-2-oxo-1,2,3,6tetrahydropyrinidine hydrochloride (Scheme 2; Step G) A mixture of (+)-6-(3,4-difluorophenyl)-1,6 methoxycarbonyl-4-methoxymethyl-l-(4-nitro)phenoxy carbonyl-2-pyrimidone (50 mg, 0.107 mmol), cis-2- [4cyano-4-(2-pyridyl)cyclohexylamino] ethylamine (40 mg, 0.164 mmol) and methylene chloride (4 mL) was stirred at room temperature overnight. After being washed with iN NaOH solution, the organic layer was concentrated and purified by preparative TLC (eluent: 10/1 ethyl acetate/2M ammonia in methanol) to give the title compound in 50% yield (31 mg) as a yellow oil: CIMS, m/z a 583 Treatment of the free base with 2 equivalents of iM HC1 in ether gave the HC1 salt as an SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 97/ 7969PCTIUS96/18573 -52off-white solid: m.p. 148-1523C. Faj 110.? 16 mg/mL MeOH). Anal. Caic. for IF' 2 N.O.2~l0.5c C, 48.58; H, 4.76; N, 11.46. Found: C, 48.30; 4.87; N, 11.20.

Example cis 3 ,4-Dif luorophenyl) 3 4 -cyalo-4.pheny..

cyclohexylaminio) propyll)Icarboxamido- 4 -methyl-2-oxo-1,2,3,6tetrahydropyrimidin hydrochloride a) General procedure for 3 -[4-Cyano.4 arylcycl1ohexylamino) propylanmine.

a) 3- 4 -Cyano-4-phenxylcycloh,,Wlamino] prepylamime (Scheme 6; Step A) A mixture of 4 -cyano-4-phenylcyclohexanone (5.0 g, 25.09 mmol) and l.

3 -diaminopropane (5.58 g, 75.3 mmcl) and p-toluenesulfonic acid (23 mg) in benzene (50 mL) was ref luxed for 4 h in Dean-Stark trap to remove the water that formed. Solvent was evaporated and the residue was redissolved in methanol (40 mL) and cooled to 0 To this, sodium borohydride (6.4 5 g) was added in portions and the mixture was stirred at room temperature for 3 h. Solvent was evaporated, the residue was dissolved in dichjloromethane (300 mL) washed with brine (3 X 500 mt), dried (potassium carbonate) and the solvent evaporated to leave the product as a pale yellow viscous oil (4.5 g, 72's). The 'H-Nl'R showed this product to be pure and found to contain the cis/trans isomers in the ratio of about 4:1.

b) cis-6- 3 ,4-Difluorophenyl) [3 (4cyano-4 p h 0n y 1 cyclohexylamino) propyll }carboxamido-5 methoxycarbonyl 4 3thyl.

2 oxo1,2,3,6-.ttrahydropyrimidine SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -53hydrochloride (Scheme 6; Step B).

A solution of 5-methoxycarbonyl-4-methyl- 2 ,3,C tetrahydro-2-oxo-6-(3,4-difluorophenyl)--[(4nitrophenyloxy) carbonyllpyrimidine (0.500 g, 1.15 n~ml) and 3- 4-cyano-4-phenylcyclohexylaminopropy (0.371 g, 1.49 nmol) in dichioromethane (50 mL) was stirred at room temperature for 12 hours, it was washed with ice-cold 0.5N NaOH (2 X 10 mL) and dried over sodium sulfate. Solvent was evaporated at reduced pressure and the residue was purified by flash chromatography on silica gel (dichloromethane:MeH.:2M ammonia in MeOH, 90:8:4) to give 0.570 g (87.6) of the product as a white powder. The HCl salt was prepared by treatment of a solution of the free base in ether with 1N HCl in ether. M.p. 163-165 OC; Anal. Calcd. for

C

3 0H 34 NsO4F 2 Cl'l.2 H 2 0:C, 57.77; H, 5.88; N, 11.23. Found: C, 57.65; H, 5.71; N, 11.20.

Example 16 3-((4-(3,4-Difluorophnyl)2,di 1,2,3, 45,7'hexahydro-4H.furo(3, 4-dI-pyrimidine-3 carbonyll amino) -ethyl- 4 -cyano-4-phenyleyclohexl) amine hydrochloride.

a) (+)-6-(3,4-Difluorophlenyl)-1,6-dihydro-2-oxo-5.

methozy-crboyl-4bromomthy1.l.C(4-nitrophonyloxy) carbonyll pyrimidine (Scheme 7; Step A).

To a well stirred solution of difluorophenyl)-1,6-dihydro-2-methoxy-5methoxycarbonyl-4-methyl-l-[(4-nitrophenyloxy) carbonyllpyrimidine (1.5 mmol, 0.66 g) in 5 mL of chloroform was added a solution of bromine (1.5 mmol, 0.09 mL) in 3 mL of chloroform at 0 0 C and the solution was allowed to attain room temperature over 1.5 h. The solvent was removed in vacuo and the residue was again dissolved in CHC1 3 (20 mL) and washed with brine. The organic layer was separated, dried over NaSO, filtered SUB STITUTE S H EET (RU LE 26) WO 97/17969 PCT/US96/18573 -54and the solvent was removed in vacuo to get 0..81 g of (-)-6-3,4-difluorophenyl)-1,6-dihydro- 2-oo methoxycarbonyl-4-bromomethy -1-[(4-nitroPhenyloxy) carbonylpyrimidine as a yellow foam. It was used in the next step without any purificatio..

b) 4 -(3,4-Difluorophnyl)-,5.diaoxo.1,2, 4 ,5,6,7hexahydro-cyclopeztapyrimidine.3-carboxylic acid-4nitrophenyl ester (Scheme 7; Step B).

(+)-6-(3,4-Difluorophenyl) 1,6-dihydro-2oxo-5etoxy.

carbonyl-4-bromomethyl-l- [(4-nitrophenyloxy)carbonyll pyrimidine (1.5 mmol, 0.81 g) was heated in oil bath for 3 h (bath temperature 130 0 C. The brown residue thus obtained was washed with CHCl 3 and (3,4-dif luorop he nyl) 2 5 d i oxo 1 2 4 5 6 7- h e x a h y d r o cyclopentapyrimidine-3-carboxylic acid-4-nitrophenyl ester was obtained as a pale brown solid which was used in the next step without further purification (crude wt. O.Slg).

C) 4-Difluorophonyl)-2,5-dioo- 1, 2 ,3,4,S.7-hexahydo-4x-furoC3,4-dJpyrimidine-3carbonyl] amino)-ethyl- (4-cyano-4 -phenylcyclohexyl) amine hydrochloride (Scheme 7; Step C).

A solution of (+)-4-(3,4-difluorophenyl)-2,5-dioxo- 1,2,4,5,6, 7 -hexahydro-cyclopenta pyrimidine-3carboxylic acid-4-nitrophenyl ester (86 mg, 0.20 mmcl) and cis-2- 4 -cyano-4-phenylcyclohexylamino) ethyl amine (70 mg, 0.3 mmol) in THF (5 m) was stirred at room temperature for 12 hours. It was purified by preparative tlc on silica gel (ethyl acetate/methanol, 90:10) to give 91 mg as a white powder. The HCl salt was prepared by treatment of a solution of the free base in ether with iN HCl in ether. [a] 0 +102 (c 0.75, MeOH); Anal. Calcd. for C 28

H

27

NSO

4 F2C1O .38CH 2 Cl 2 C, 56.41; H, 4.80; N, 11.59. Found: C, 56.80; H, 4.96; N, 11.29.

SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 Example 17 (+)-cis-6-(3,4-Difluorophenyl) 4-( 4 -cyano-4-phenylcyclohexylamino)butyll 5 -methoxycarbonyl 2, 4-dimethyl 1,6-dihydropyrimidine hydrochloride (Scheme 8).

To a suspension of NaH (50 mg, 60% dispersiocn in mineral oil) in THF (7 mtL) was added a solution of 6- (3,4-difluorophenyl)-1, 6 -dihydro-5-methoxycarbonyl-2,4dimethylpyrimidine (0.32 g, 1.14 mmol.) and HMPA (0.205g, 1.14 mmol.) in THF (8 mL) at OOC. After min, 1,4-dibromobutane (0.47 mtL, 4.0 mmol.) was added.

The reaction mixture was then refluxed for 10 min. The solid was filtered off. After the removal of the solvent, the residue was flash chromatographed over silica gel (eluent: ethyl acetate) to give the product in 44% yield (0.2 g) as a yellow oil.

A mixture of 1-(4-bromobutyl)-6-(3,4-difluorophenyl)- 1,6-dihydro-5-methoxycarbonyl-2, 4 -dimethylpyrimidine (0.1 g, 0.43 mmol), cis-4-cyano-4-phenylcyclohexylamine (0.18 g, 0.9 mmol), potassium carbonate (0.3 g, 2.1 mmol), sodium iodide (65 mg, 0.43 mmol) and acetone (6 mL) was refluxed overnight. After removal of the solid and solvent, the residue was purified by preparative TLC (eluent: 100/20 v/v ethyl acetate-2M ammonia in methanol) to give the product in quantitative yield (0.23 g) as a pair of diastereomers. HPLC separation (Column: Chiralcel *OD 20x250 mm; Eluent: 60/40/0.1 hexane/2-propanol/diethylamine) gave the pure title compound as a yellow oil: DCIMS, m/z 535 (MW) Treatment of the free base with 2 equivalents of 1M HCI in ether gave the HC1 salt as a white solid: m.p. 169- 172 0 C. [ar 68.6 (2.2 mg/mL MeOH). Anal. Calc. for

C

3 iHF 2

N

4 0, 2 2HC1 0.9CMC1 3 C, 53.59; H, 5.48; N, 7.84.

Found: C, 53.61; H, 5.67; N, 7.81.

It is understood that additional compounds may be synthesized under the general synthetic schemes SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -56referred to herein using appropriately substituted starting materials.

Example 18 (+)-6-(3,4-Difluorophenyl)-l-{N-[2-(4-cyano-4phenylcyclohex- 1-ylamino) -ethyl] carboxamido-5 -methoxycarbonyl- 4-hydroxymethyl-2-oxo-1,2,3, -tetrahydropy-rimidine To well-stirred solution of (+)-6-(3,4-difluorophenyl)-1.NN[ 2 4 -cyano-4-phenylcyclohex-1.ylaino)-ethyl] carboxamido- S-methoxycarbonyl-4-methoxymethyl-2-oxo- 1,2, 3 ,G-tetrahydropyrimidine (0.108 g, 0.184 mmol) in dichioromethane (8 mL) at -780C was added a 1M solution of boron tribromide (0.55 mL) dropwise and the cooling bath was removed.

After stirring the mixture at room temperature for 4 h, it was quenched by the addition of methanol (5 mL). Solvent was evaporated and the residue was purified by preparative thinlayer chromatography on silica gel using chloroform/methanol/2M ammonia in methanol (95/4/1) as the eluent. The faster moving component (Rf 0.6, TLC on silica gel plates eluting with 25:2:1 CHC1 3

:CH

3 0: 2M NH 3 in CH 3 OH) corresponded to 3 ,4-difluorophenyl) [2-(4-cyano-4phenylcyclohex-1-ylamino) -ethyl] carboxamido-5-methoxcarbonyl 4 bromomethyl-2-oxo1,2,3,S-tetrahydropyrimidine and it was converted to the hydrochloride salt by treatment with iN HCl in ether (10 mg); MH+ 630; M.P. 225-227 OC; [a]D +126 (c 0.115, MeOH); Anal. Calcd. for C 29

H

31

NO

4 FC1.0.2CH 2 C1. C, 51.28; H, 4.63; N, 10.24. Found: C, 51.40; H; 4.58; N, 10.12. A second component (Rf 0.5, TLC on silica gel plates eluting with 25:2:1 CHC1 3

CH

3 OH: 2M NH 3 in CH3OH) was identified as 3 4-difluorophenyl)-l-{N-[2-(4 -cyano-4-phenylcyclohex-1ylamino) -ethyl]carboxamido-5-methoxycarbonyl.4-hydrox ymethyl-2-oxo-1,2,3,6-tetrahydropyrimidine (45 mg); MH 568; M.P. 230-232 OC; £a]D +123 (c 0.275, MeOH); Anal. Calcd. for

C

29

H

3 lNO 5

F

2 .0.35CH 2

C

2 C, 59.02; H, 5.35; N, 11.72. Found: C, 58.93; H; 5.50; N, 11.68.

SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCTIUS96/18573 -57- Example 19 3 ,4-Difluorophenyl)-l-{N-[2-(4-cyano-4phenylcyclohex--ylamino)-ethyl]}carboxamido-4-methoxymethyl-2cxc 1,2,3,6-tetrahydopyrimidne-5-carboxylic acid.

a) Benzyl 3 ,4-difluoropheny1)methylene-3-XO-4meihoxybutyrate. To a solution of benzyl 4 -methoxyacetoacetate (84 g, 0.37 mci), 3 ,4-difluorobenzaldehyde (52.5 g, 0.0.37 mmcl), and piperidinium acetate (1.58 g, 2.63 mmol) in benzene (1L) were added molecular sieves (400 g) and the mixture was stirred at room temperature for 48 h. The molecular sieves were removed by filtration and the solvent was evaporated from the filtrate under reduced pressure. The residue was purified by column chromatography on silica gel using chloroform/ethyl acetate (100:3) to get the product as a white powder (118 g, The NMR showed this product to be a mixture of cis and trans isomers.

b) 5-Benzyloxycarbonyl-1,6-dihydro-2-methoxy-4-methoxymethyl-6- (3, 4 -difluorophenyl)pyrimidine. A suspension of benzyl difluorophenyl)methylene 3 -oxo-4-methoxybutyrate (60 g, 0.181 mol), O-methylisourea hemisulfate (46.77 g, 0.271 mol, 1.5 eq.), and sodium bicarbonate (53.25 g, 0.634 mol, 3.5 eq.) in ethanol (400 mL) was stirred at 50-50 C for 6 h. The solvent was evaporated from the combined filtrates and the residue was purified by column chromatography (SiO 2 EtOAc/hexane, 10%-30%) to give 5-benzyloxycarbonyl-1,6-dihydro-2-methoxy-4methoxymethyl-6-(3,4-difluorophenyl)pyrimidine as a pale yellow oil (24.7 g, c) 5-Benzyloxycarbonyl-4-methoxym ethyl-1,6-dihydro-2-methoxy-6- (3, 4 -difluorophenyl) -nitrophenyloxy) carbonyl] pyrimidine.

To a well-stirred solution of S-benzyloxycarbonyl-1.6-dihydro-2methoxy-4 4methoymethyl-6-(3, 4 -difluorophenyl)pyrimidine (12 g, 29 mmcl) and pyridine (10 mL) in CH 2 C1 2 (100 mL) was added powdered 4 -nitrophenyl chioroformate (8.69 g, 43 mmol) at OOC.

The reaction mixture was stirred for 12 h at room temperature and SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 9717969PCT/US96/1 8573 -58the solvent was evaporated from the filtrate and the residue was purif ied by column chromatography (Si0 2 dichioromethane/hexane, 20oi-500i) to give the product as a foam (14.2 g, 86*).

d) -S-Benzyloxycarbonyl.4-.methoxymethy16dhdr-{ [2phenyl) ethyl] }carboxamido-2-methoxy 6 difluorophenyl)pyrimidine. To a stirred solution of benzyloxycarbonyl..4-.methoxmethyl 1 -dihydro-2-methoxy- 6 difluorophenyl) (4 -nitrophenyloxy) carbonyl] pyrimidine (6 g, 10.5 mmol) in THF (120 mL) was added R-(+)-a-methylbenzylamine (1.8 g, 15 mmol) at room temperature and the stirring was continued for 12 h. Solvent was evaporated and the residue was purified by column chromatography (SiO., 10-2001 EtOAc in hexane).

The first major product to elute was (+)-5-benzyloxycarbonyl.

4 methoxmethy.i, 6-dihydro-i- [2-phenyl) ethyl]l~carboxamido.2 methoxy-6-( 3 ,4-difluorophenyl)pyrimidine as an oil.

e) -S-Benzylcxycarbonyl..i6-dihycfro-2 -methoxy-4 -methoxymethyl- 6-( 3 ,4-difluorophenyl)pyrimidine. To a stirred solution of benzyloxycarbonyl.4..methoxmethyl-1,6 -dihydro-.- (2phenyl) ethyl] }carboxamido-2.methox..E..(3,4difluorophenyl)pyrimidine (2 g, 3.5 mmol) in dich].oromethane (200 mL) was added lP 8 -diazabicyclo[5,4,0)undec.7ene (0.6 mL) and the mixture was stirred at room temperature for 12 h. Solvent was evaporated and the residue was purified by flash column chromatography on silica gel with 30-40!k EtOAc in hexane as eluent to give -S-benzyloxycarbony>l16-dihydro-2-methoxy-4methoxymethyl-6-.(3, 4 -difluorophenyl) pyrimidine as a viscous oil (1.25 g, 86U) f) -5 -Benzyloxycarbonyl.4 -methoxymethyl1, 6-dihydro-2-methoxY-.

6- (3,4 -difiluorophenyl) -1-[(4-nitrophenyloxyj carbonyl] pyrimidine.

To a well stirred solution of (+)-5-benzyloxycarbonyl-1,6diyr--ehx-4mtoyehl6(3,4difluorophenyl)pyrimidfle (1.25 g, 3.0 mmcl) and pyridine (2 ML) in CI{ 2 C1 2 (20 mL) was added powdered 4-nitrophenyl chioroformate, SUIBSTTUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -59- (1 g, 5 mmol) at room temperature. Solvent was evaporated and the residue was purified by column chromatography on silica gel using CHC1 3 /EtOAc as the eluent to give benzyloxycarbonyl-4-methoxymethyl-1,6-dihydro-2-methoxy-6-(3,4diflurophenyl)-1-[(4-nitrophenyloxy)carbonyl]pyrimidine (1.70 g, 86%) as a viscous oil. +200 (c 1.215 g in 100 mL CHC1 3 g) (+)-6-(3,4-Difluorophenyl)-1-{N-[2-(4-cyano-4phenylcyclohex-1-ylamino)-ethyl]}carboxamido-S-benzyloxycarbonyl- 4-methoxymethyl-2-oxo 1,2,3, 6 -tetrahydropyrimidine (cis isomer).

A solution of (+)-6-(3,4-difluorophenyl)-1,6-dihydro-2-methoxy-5benzyloxycarbonyl-4-methoxymethyl-1-(4-nitrophenoxy)carbonylpyrimidine (0.100 g) and 2-[4-cyano-4-phenylcyclohex-1yllaminoethylamine (0.05 g) in THF (5 mL) was stirred at room temperature for 12 hours. To this 6N HC1 (0.5 ml) was added and the stirring was continued for 2 h. Solvent was evaporated, the residue was mixed with dichloromethane (10 mL), washed with icecold 0.5N NaOH (2 X 10 mL) and dried over sodium sulfate.

Solvent was evaporated at reduced pressure and the residue was purified by flash chromatography on silica gel (dichloromethane:MeOH:2M ammonia in MeOH, 90:8:4) to give 0.12 g of the product as a white powder. The NMR showed this product to be pure and was used in the next step without any further characterization.

h) (+)-6-(3,4-Difluorophenyl)-1-{N-[2-(4-cyano-4phenylcyclohex-1-ylamino)-ethyl]}carboxamido-4-methoxymethyl-2oxo 1,2,3,6-tetrahydropyrimidine-5-carboxylic acid.

To a solution of (+)-6-(3,4-difluorophenyl)-l-{N-[2-(4-cyano-4- 4-methoxymethyl-2-oxo 1,2,3,6-tetrahydropyrimidine (120 mg) in methanol/water 10 mL), 10% palladium on charcoal (20 mg) was added and the mixture was hydrogenated at 125 psi for 4 h.

The tic analysis of the reaction indicated the completion of the reaction. The catalyst was removed by filtration and the solvent was evaporated to leave the product as a pale yellow powder (86 SUBSTITUTE SHEET (RULE 26) WO 97/1 7969 PCTIUS96/1 8573 mg) m.p 188-191WC; Anal. Calcd. for C 29

H

31 N,0SF, 0.5H 2 0: C, 60.41; Hl, 5.59; N, 12.15. Found: C, 60.28; H; 5.62; N, 12.34.

SUBSTiTUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -61- Example As a specific embodiment of an oral composition cf a compound of this invention, 100 mg of one of the compounds described herein is formulated with sufficient finely divided lactose to provide a total amount of 580 to 590 mg to fill a size 0 hard gel capsule.

Pharmacological Profiles of the Com-ounds in Cloned Human Adrenergic Receptors.

Binding affinities were measured for selected compounds of the invention at six cloned human alpha-1 and alpha- 2 receptor subtypes, as well as at the L-type calcium channel. The protocols for these experiments are given below.

Protocol for the Determination of the Potency of a, Antaqonists The activity of compounds at the different human receptors was determined in vitro using cultured cell lines that selectively express the receptor of interest. These cell lines were prepared by transfecting the cloned cDNA or cloned genomic DNA or constructs containing both genomic DNA and cDNA encoding the human a-adrenergic receptors as follows: a, Human Adrenergic Receptor: The entire coding region of a, (1719 bp), including 150 base pairs of untranslated sequence UT) and 300 bp of 3' untranslated sequence UT), was cloned into the BamHI and Clal sites of the polylinker-modified eukaryotic expression vector pCEXV-3, called EXJ.HR.

SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -62- The construct involved the ligation of partial overlapping human lymphocyte genomic and hippocampal cDNA clones: 5' sequence were contained on a 1.2 kb SmaI-XhoI genomic fragment (the vector-derived BamHI S site was used for subcloning instead cf the internal insert-derived Smal site) and 3' sequences were contained on an 1.3 kb XhoI-ClaI cDNA fragment (the Clal site was from the vector polylinker). Stable cell lines were obtained by cotransfection with the plasmid alA/EXJ (expression vector containing the alA receptor gene (old nomenclature)) and the plasmid pGCcos3neo (plasmid containing the aminoglycoside transferase gene) into LM(tk-) cells using calcium phosphate technique. The cells were grown, in a controlled environment (37 0 5% CO 2 as monolayers in Dulbecco's modified Eagle's Medium (GIBCO, Grand Island,

NY)

containing 25mM glucose and supplemented with bovine calf serum, 100 units/ml penicillin g, and 100 Mg/ml streptomycin sulfate. Stable clones were then selected for resistance to the antibiotic G-418 (1 mg/ml), and membranes were harvested and assayed for their ability to bind ['HIprazosin as described below (see "Radioligand Binding assays").

The cell line expressing the human receptor used herein was designated (old nomenclature) and was deposited with the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852, U.S.A.

under the provisions of the Budapest Treaty for the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

The cell line expressing the human av receptor, was accorded ATCC Accession No. CRL 11138, and was deposited on September 25, 1992.

a HRuman Adrenergic Receptor: The entire coding region of alB (1563 bp), including 200 base pairs and SUBSTITUTE SHEET (RULE 26) -63untranslated sequence UT) and 600 bp of 3' untranslated sequence UT) was cloned into the EcoRI site of pCEXV-3 eukaryotic expression vector.

The construct involved ligating the full-length containing EcoRI brainstem cDNA fragment from X ZapII into the expression vector. Stable cell lines were selected as described above. The cell line used herein was designated L-a B and was deposited with the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852, U.S.A. under the provisions of the Budapest Treaty for the International Recognition of the Deposit of Microorganisms for the SPurposes of Patent Procedure. The cell line L-alB was accorded ATCC Accession No. CRL 11139, on September 1992.

aA Human Adrenergic Receptor: The entire coding region of (1401 bp) including 400 base pairs of untranslated sequence UT) and 200 bp of 3' S 20 untranslated sequence UT), was cloned into the KpnI site of the polylinker-modified pCEXV-3-derived eukaryotic expression vector, EXJ.RH. The construct involved ligating three partial overlapping fragments: a 5' 0.6kb HincII genomic clone, a central 1.8 EcoRI hippocampal cDNA clone, and 3' 0.6Kb PstI genomic clone. The hippocampal cDNA fragment overlaps with the and 3' genomic clones so that the HincII and PstI sites at the 5' and 3' ends of the cDNA clone, respectively, were utilized for ligation. This fulllength clone was cloned into the KpnI site of the expressed vector, using the 5' and 3' KpnI sites of the fragment, derived from vector pBluescript) and 3'-untranslated sequences, respectively. Stable cell lines were selected as described above. The stable cell line expressing the human a, receptor used Sherein was designated L-aic (old nomenclature) and was deposited with the American Type Culture Collection, fc 5 dpstdwt h mrcnTp utr olcin -64- 12301 Parklawn Drive, Rockville, Maryland 20852, U.S.A.

under the provisions of the Budapest Treaty for the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

The cell line expressing the human CIA receptor was accorded Accession No. CRL 11140, on September 1992.

Radioligand Binding Assays: Transfected cells from culture flasks were scraped into 5ml of 5mM Tris-HCl, 5mM EDTA, pH 7.5, and lysed by sonication. The cell lysates were centrifuged at 1000 rpm for 5 min at 4 0

C,

and the supernatant was centrifuged at 30,000 x g for 20 min at 4 0 C. The pellet was suspended in 50mM Tris- HCL, ImM MgCL 2 and 0.1% ascorbic acid at pH Binding of the al antagonist 3 H]prazosin (0.5 nM, specific activity 76.2 Ci/mmol) to membrane preparations of LM(tk-) cells was done in a final volume of 0.25 ml and incubated at 37 0 C for 20 min.

Nonspecific binding was determined in the presence of 20 10 lM phentolamine. The reaction was stopped by •filtration through GF/B filters using a cell harvester.

Inhibition experiments, routinely consisting of 7 concentrations of the tested compounds, were analyzed using a none-linear regression curve-fitting computer program to obtain Ki valves.

a 2 Human Adrenergic Receptors: To determine the potency of al antagonists at the 0 2 receptors, LM(tk-) cell lines stably transfected with the genes encoding the

C)

2 A, C2B, and a2c receptors were used. The cell line expressing the a2A receptor is designated L-C2A, and was deposited on November 6, 1992 under ATCC Accession No.

CRL 11180. The cell line expressing the (2B receptor is designated L-NGC-C 2 B, and was deposited on October 1989 under ATCC Accession No. CRL 10275. The cell line g, expressing the Q2c receptor is designated L-2c, and was deposited on November 6, 1992 under ATCC Accession No.

WO 97/17969 PCT/US96/18573 CRL-11181. All the cell lines were deposited with the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852, U.S.A. under the provisions of the Budapest Treaty for the International S Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. Cell lysates were prepared as described above (see Radioligand Binding Assays), and suspended in 25mM glycylglycine buffer (pH 7.6 at room temperature). Equilibrium competition binding assay were performed using [3H]rauwolscine and nonspecific binding was determined by incubation with 10gM phentolamine. The bound radioligand was separated by filtration through GF/B filters using a cell harvester.

Determination of the Activity of a, Antagonists at Calcium Channels The potency of a, antagonists at calcium channels was determined in competition binding assays of [PHnitrendipine to membrane fragments of rat cardiac muscle, essentially as described by Glossman and Ferry (Methods in Enzymology 109:513-550, 1985). Briefly, the tissue was minced and homogenized in 50mM Tris-HC1 (pH 7.4) containing 0.1mM phenylmethylsulfonyl fluoride. The homogenates were centrifuged at 1000 g for 15 minutes, the resulting supernatant was centrifuged at 45,000 g for 15 minutes. The 45,000 g pellet was suspended in buffer and centrifuged a second time. Aliquots of membrane protein were incubated for 30 minutes at 37C in the presence of P'Hnitrendipine (InM), and nonspecific binding was determined in the presence of 10gM nifedipine. The bound radioligand was separated by filtration through GF/B filters using a cell harvester.

The compounds described above were assayed using cloned human alpha adrenergic receptors and the rat calcium SUBSTITUTE SHEET (RULE 26) WO 97/1 7969 PCTIUS96/1 8573 -66channel. The preferred compounds were found to be 1 selective antagonists. The compounds described herein were also found to exhibit weak binding affinity to, the rat L-type calcium channel. The alpha adrenergjc receptor binding affinit-ies of compounds 2-5, 7, 12., 13 and 14 are illustrated in Table I. as follows: Table 1 Binding affinities of selected compounds at cloned human alpha-. adrenergic receptors Example haID hals Exampleha -a~h 1 I A SEM n pl( 1 SEM n pK 1 SEM n 2 7.05 0.07 2 7.76 0.07 2 9.51 0.07 2 3 6.62 0.02 3 7.00 0.03 3 9.31. 0.07 3 4 7.00 0.10 3 7.73 0.07 3 9.87 0.07 4 6.91 0.10 3 7.57 0.06 3 9.90 0.01 4 7 5.41 0.03 2 5.95 0.09 2 8.35 0.24 2 11 6.90 0.04 2 7.56 0.03 2 9.66 0.11 2 13 7.29 0.02 2 7.87 0.01. 2 9.99 0.18 2 14 5.96 0.04 2 6.55 0.04 2 9.03 0.08 2 SUBSTITUTE SHEET (RULE 26) WO 97/1 7969 PCT/US96/1 8573 -67- Part i P. 4l 1 PIPS 2. NA S H

A

p R,

R

Part 2 y

C

y2 y3

RR-B

TH2 H NR

C

>QN 0 C~ 0 (vbon Rl".U4o)

D

y 2

R

R

HC I

E

Part- 3 ft a SCHEME 1 a. p-toluen*auulfnic acid SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -68- In Part 1 of the general synthetic scheme, an optionally substituted cycloalkanone is refluxed with a diaminoalkane to form the corresponding (cycloalk-lyl)aminoalkylimine, which on treatment with NaBO, yields (cycloalk-1-yl) aminoalkyiamine. In part 3 the diamine is added to a 1-[(4-nitrophenyl)oxycarbonyl]pyrimidine to yield the final product. Although cyclohexanone is shown, the process is equally applicable to substituted cycloalkanones from cyclobutanone through cyclooctanone. In addition, replacement of the diaminoalkane of Part 1 with an aminoalkanol yields esters in Part 3 instead of amides. In the schemes which follow, the diaminoalkane is shown for clarity as either ethyl, propyl or butyl; however, any suitable alkyl, branched alkyl or substituted alkyl as shown in Scheme 1 may be used, wherein R is as defined in the specification.

In Part 2 of the general synthetic scheme, it is important to note that the use of the O-methylisourea OMe) in Step D allows the production of several different 2-substituents on the dihydropyrimidine. In particular, addition of primary amines NH2R 3 to the 2-methoxy pyrimidines affords 2-amino pyrimidines.

Alternatively, when OMe is replaced with 4methoxybenzylthiol, 2 -thioxodihydropyrimidines may be produced by treatment of the product with ethanethiol and TFA.

SUBSTITUTE SHEET (RULE 26) WO 97/1 7969 PCT/US96/18573 -69- P4gt-I H 4

A:

R Benlenq 2.

H-

4

_OA

c Ph meyidl 2~ N C H 250o" HoOH Part 2 CHO0 y y 0 R

R

NH

m N y *t 0 N R Cl 0 Iwhon 1--og4.) it..

Paz t- As I-h.2pyrilyl. 2-Pfb Y2 Ip R' Me. tt.MOeOCH 2 a- H. CH.CO2 e Ph 1*1 Mo, w SOWIE 2 4.Ph phonyl b. n benayl S UB STITUTE S HE ET (RU LE 26) WO 97/17969 WO 9717969PCTIUS96/18573 y y2 0 It N Ome 0 C I 0

B

y Y 3

N

0 Metbod 1 WPtoC aeparation R' N ONe 1. ft(*)-Phenethylamine 2. Sopazation of diaaserson C 3DBU RO N 1e ()and ernners SCHE 3 a. DDO 1.S-diazabcclo(S.g.oIundec7.en SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 9717969PCT/US96/18573 -71--

'ILI

N om EriO N 0 N c H c'C H N Ph H C N 2. H 2/Pd-C i. ZCD .DKAP .BHI

B

SCHDME 4 A.*C 1- Diathylaainvxopyl) thylcazbodiimid. hydrochder ido b.N 4-Diathylamifo~?±in SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 -72- 2Ph 2 K prs Sensene NM )4c Ph a 2

=-S

m o N Pb r "a me U 0 2. DN 4

N

k* N 0 ,ft ci isK Cl*WI 08 m0 Het wL 9 2 co) 0 N K K

N.

I

tram 1 1 CKMIP AO xmple 20 SMum4E SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 9717969PCTIUS96/18573 -73- H N NH Ph 2 2 O C I1. PTS Denzene p 2. NaBH m2Nl 4 me 0

N

H

Ph 8 r

F

CN

SCHEME 6 SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/1 8573 -74-

KF

Me N Nmo r' NO2 Me 0N BrCH N 0

H

~130 *C

B

o 2O H P N qw\

X%

H

2

N

C

00 N O H H

CN

SCHEME '7 SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573

F

F

me 0 NH u- N a.

N&H, HMPA

A

Br He N He IPh

HC

B W K D :0 Mal

F

H HK Ph MOO N Hw CN Mo N He SCHEME 8 a. A hexamuthylphosphozaamide SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 9717969PCT/US96/1 8573 -76c1zJ z 4

U-=

\Z

r.4 ca I L C)

Z

SUBSTITUTE SHEET (RULE 26) WO 97/1 7969 PCT/US96/18573 -77z z u.

-2 z 7 2

C

0

N

II

II

z

C

La 2 2 Lx..

C

N

u

T-

N

0 C

ON

2 z 0

Z=

u O 0 SUBSTITUTE SHEET (RULE 26) WO 97/1 7969 PCT/US96/18573 -78- Similar compounds may be produced by substitution of the 1,4-dibromobutane with other dibromoalkanes, including branched compounds in order to produce the alkyl-linked cycloalkylamine derivatives of the subject dihydropyrimidines.

SUBSTITUTE SHEET (RULE 26)

Claims (40)

1. A compound having the structure: if I R 3 9 I R 2 SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 97/ 7969PCT/US96/18573 x N R 3 Li R 4 R B or SUBSTITUTE SHEET (RULE 26) VO 97/17969 WO 9717969PCTIUS96/1 8573 R N R 2 where M has the structure .Y 3 Yi 1 Y 4 wherein each of Y 1 ,1 Y 2 Y 3 1 Y, and Y. independently may be -H; straight chained or branched Cl-C., alkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 -C 7 alkenyl or alkynyl; C 3 -C 7 cycloalkyl, monofluorocycloalkyl, polyfluorocycloalkyl or cycloalkenyl; -Cl, -Br, or -I; -NO 2 -N(R 3 2 -N 3 -CN; -OR 3 -OCOR 3 -COR 3 -CON(R 3 2 or SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 82 -CO.,R 3 or wherein two of -Y 1 Y n 5 aepeeto adjacent carbon atoms and together constitute a methylenedioxy group; where R independently may be straight chained or branched alkyl., monof2.uoroalkyl or polyfluoroalkyl; straight chained or branched C 2 alkenyl or alkynyl; -N(R 3 2 -NO 2 -CN; -C0 2 R 3 (CH 2 PPO(OR 3 2 (CH 2 PPO(O; or R3 where R, may be -NO 2 -CN; straight chained or branched c 1 alkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 alkenyl or alkynyl; C 3 cycloalkyl, mono fluorocyc loalkyl, polyfluorocycloalkyl or cycloalkenyl; -N(R 3 2 -OR 3 (CHI.) P OR 3 -COR 3 -C0 2 R 3 or -CON(R 3 2 1 wherein any p independently is an integer from 2. to 7 inclusive; where R 2 may be straight chained or branched C1-C, alkyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 alkenyl or alkynyl; C 3 -C7 cycloal-kyl, monoflu 'orocycloalkyl, polyfluorocycloalkyl or cycloalkenyl; C-I cycloalkyl-Cl-Cl 0 -alkyl, C 3 -C, 0 cycloalkyl-C,-C, 1 0 -monof luoroalkyl or C 3 -C11 0 cycloalkyl-Cl-Cl.-polyf luoroalkyl; -CN; CH 2 XR 3 ,I CH 2 X (CHO) NHR 3 (CH 2 ,NHR 3 -CH 2 X(CH 2 N (R 3 2 1 -CH 2 X (CH 2 PN 3 1 or -CH 2 X (CHO NHCXR.7 or -OR 3 wherein any n independently is an integer f rom 0 to inclusive and p is as defined above; where each R 3 independently may be straight chained or branched C 7 alkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 -C 7 alkenyl or alkynyl; C 3 -C 7 SUBSTITUTE SHEET (RULE 26) VO 97/17969 WO 97/ 7969PCT/US96/18573 83 cycloalkyl, ronofjluorocycloalkyl, polyfluorocycloalkyl or cycloalkenyl; (CH 2 ,PO (ORB) 2 (CH 2 PPO (ORB) 3 or (CH 2 PCO0R9; where R 4 has the structure SI R 6 wherein Z may be C 2 alkenyl or alkynyl; CH 2 0; CO; CO 2 CONR 3 CO; CONR 3 S; SO; SO 2 or NR 3 m is an integer f rom 0 to 3 inclusive; R. and R, each independently may be -Cl, -Br, -I; -C0 2 R 3 -COR 3 -CON(R 3 -CN; -NO 2 -N(R 3 2 -OR 3 straight chained or branched Cl-C, alkyl, Cl-C, mono fluoroal kyl, C 1 -C7 polyfluoroalkyl, C 2 -C-7 alkenyl, C 2 -CI alkynyl, C 3 -C-7 cycloalkyl, or C 3 cycloaJlkenyl, wherein the al~kyl, mono fluoroalkyl, polyfluoroalkyl, alkenyl, alkynyl, cycloalkyl or cycloalkenyl may be unsubstituted or substituted with -H, aryl or heteroaryl; aryl. or heteroaryl wherein the aryl or heteroaryl may be unsubstituted or substituted with -F, -Cl, -Br, -NO 2 -CN, -N(R 3 2 -OR 3 -COR 3 -C0 2 R 3 or -CON (RO) 2 straight chained or branched Cl-C7 alkyl, rnonofluoroaJlkyl or polyfluoroalkyl, straight chained or branched C2 alkeny. or alkynyl, C 3 -C 7 cycloalkyl, monof luorocycloalkyl, polyf luorocycloalkyl, or cycloalkenyl; R, may be -ti; straight chained or branched Cl-C 7 alkyl, hydroxyalkyl, aminoalkyl, alkoxyalkyl, monofluoroalkyl or SUBS1TrLTE SHEET (RULE 26) W O 97/17969 PCT/US96/1 8573 84 p.olyfluoroalkyl; straight chained or branched C 2 alkenyl or alkynyl; C 3 cycloalkyl, monofluorocycloalkyl, polyflucrocycloalkyl or cycloalkenyl; -CN; -C0 2 R 3 -CQR 3 -CON(R 3 2 or -OR 3 where R, is straight chained or branched C,-C 1 alkyl, monofjluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 -C 7 alkenyl or alkynyl; C 3 C 7 cycloalkyl, monofluorocycloalkyl, polyfluorocycloalkyl or cycloalkenyl; where R 9 may be -CN; straight chained or branched C 1 alkyl, monof luoroalkyl or polyf luoroalkyl; straight chained or branched C 2 -C 7 alkenyl or alkynyl; C 3 -C 7 cycloalkyl, mono fluorocyc loalkyl, polyfluorocycloalkyl or cycloajlkenyl; -NCR 3 2 -OR 3 -(CH 2 )p OR 3 -COR 3 -C0 2 or -CON(R 3 2 wherein X may be S; 0; or NR,; and wherein 2 may be straight chained or branched C 1 -C 7 alkyl, ronofluoroalkyl, polyfluoroalkyl, alkoxy or thioalkyl; straight chained or branched C 2 alkenyl; -SCH 2 C 6 H 4 OR 3 (CH 2 nC6H 5 CH 2 X (CHO,1NHR 3 (CH 2 NHiR 3 or OR 3 or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein the compound comprises the enantiomer.

3. The compound of claim 1, wherein the compound comprises the enantiomer. SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573

4. The compound of claim 1, wherein the compound is selected from the group consisting of: P R 1 /R 4 R I\ ,R N x I R3 and SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 9717969PCT/US96/18573 Ra compound of claim 4, wherein R, is 1 R and p is 2, 3 or 4. SUBSTITUTE SHEET (RULE 26) -4 C 87

6. The compound of claim 5, wherein R, is 0R R3 I RS P R. R 2 A H

8. The compound of claim 7, wherein M is polyf luorophenyl and R, is substituted or unsubstituted phenyl or pyridyl.

9. The compound of claim 8, wherein R, is -CO 2 CH 3 -COC 3 or -CONU 2 R 2 is -CHOCH,, methyl or ethyl; and R. is H, -CN, or *CO2CH 3 WO 97/17969 PCT/US96/1 8573 88 The compound of claim 6, wherein the compound is selected from the group consisting of: N.a .00 SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 97/ 7969PCTIUS96/1 8573 89 F 0 0 H2H ~~0 4w p SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 9717969PCT/US96/18573 H 'NI- 0 I .0 .0 SUBSTITUTE SHEET (RULE 26) WO 97/17969 WO 9717969PCT/US96/18573 Ce and *N% 0 ,0

11. The compound of claim 10, wherein the compound is SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 92 the enantiomer.

12. The compound of claim 10, wherein the compound is the enantiomer.

13. The compound of claim 6, having the structure: X I 0 R R R

14. The compound of claim 13, wherein M is polyfluorophenyl and R s is substituted or unsubstituted phenyl or pyridyl. The compound of claim 14, wherein the compound is: F o 0 H C 0 H

16. The compound of claim 15, wherein the compound is the enantiomer.

17. The compound of claim 15, wherein the compound is the enantiomer.

18. A pharmaceutical composition comprising a therapeutically effective amount of the compound of claim 1 and a SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 93 pharmaceutically acceptable carrier.

19. The pharmaceutical composition of claim 18, wherein the amount of the compound is an amount from about 0.01 mg to about 500 mg. The pharmaceutical composition of claim 19, wherein the amount of the compound is from about 0.1 mg to about 60 mg.

21. The pharmaceutical composition of claim 20, wherein the amount of the compound is from about 1 mg to about 20 mg.

22. The pharmaceutical composition of claim 18, wherein the carrier is a liquid and the composition is a solution.

23. The pharmaceutical composition of claim 18, wherein the carrier is a solid and the composition is a tablet.

24. The pharmaceutical composition of claim 18, wherein the carrier is a gel and the composition is a suppository. The pharmaceutical composition of claim 18, wherein the compound is formulated as a part of a pharmaceutically acceptable transdermal patch.

26. A method of treating a subject suffering from benign prostatic hyperplasia which comprises administering to the subject an amount of the compound of claim 1 effective to treat benign prostatic hyperplasia.

27. A method of claim 26, wherein the compound additionally does not cause a fall in blood pressure at dosages effective to alleviate benign prostatic hyperplasia.

28. The method of claim 27, wherein the compound effects treatment SUBSTITUTE SHEET (RULE 26) S WO 97/17969 PCT/US96/18573 94 of benign prostatic hyperplasia by relaxing lower urinary tract tissue.

29. The method of claim 28, wherein the lower urinary tract tissue is prostatic smooth muscle. A method of treating a subject suffering from elevated intraocular pressure which comprises administering to the subject an amount of the compound of claim 1 effective to lower intraocular pressure.

31. A method of treating a subject suffering from a disorder associated with elevated blood cholesterol which comprises administering to the subject an amount of the compound of claim 1 effective to inhibit cholesterol synthesis.

32. A method of treating a disease which is susceptible to treatment by antagonism of the a1A receptor which comprises administering to the subject an amount of the compound of claim 1 effective to treat the disease.

33. A method of treating a subject suffering from impotency which comprises administering to the subject an amount of the compound of claim 1 effective to treat impotency.

34. A method of treating a subject suffering from sympathetically mediated pain which comprises administering to the subject an amount of the compound of claim 1 effective to treat sympathetically mediated pain. A method of treating a subject suffering from cardiac arrhythmia which comprises administering to the subject an amount of the compound of claim 1 effective to treat cardiac arrhythmia. SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573

36. A method of treating a subject suffering from benign prostatic hyperplasia which comprises administering to the subject the compound of claim 1 in combination with a 5 alpha-reductase inhibitor in an amount effective to treat benign prostatic hyperplasia.

37. The method of claim 36, wherein the 5-alpha reductase inhibitor is finasteride.

38. A pharmaceutical composition comprising a therapeutically effective amount of a combination of the compound of claim 1 and finasteride and a pharmaceutically acceptable carrier.

39. The pharmaceutical composition of claim 38, comprising from about 0.01 mg to about 500 mg of the compound and about 5 mg finasteride. The pharmaceutical composition of claim 38, comprising from about 0.1 mg to about 60 mg of the compound and about 5 mg finasteride.

41. The pharmaceutical composition of claim 38, comprising from about 1 mg to about 20 mg of the compound and about 5 mg finasteride.

42. A method of relaxing lower urinary tract tissue which comprises contacting lower urinary tract tissue with an amount of the compound of claim 1 effective to relax lower urinary tract tissue.

43. The method of claim 42, wherein the lower urinary tract tissue is prostatic smooth muscle.

44. The method of claim 43, wherein the compound additionally does not cause a fall in blood pressure. SUBSTITUTE SHEET (RULE 26) WO 97/17969 PCT/US96/18573 96 A method of relaxing lower urinary tract tissue in a subject, which comprises administering to the subject an amount of the compound of claim 1 effective to relax lower urinary tract tissue.

46. The method of claim 45, wherein the lower urinary tract tissue is prostatic smooth muscle.

47. The method of claim 46, wherein the compound additionally does not cause a fall in blood pressure.

48. A method of inhibiting contraction of prostatic tissue, which comprises administering to the subject an amount of the compound of claim 1 effective to inhibit contraction of prostatic tissue.

49. The method of claim 48, wherein the prostatic tissue is prostatic smooth muscle. The method of claim 49, wherein the compound additionally does not cause a fall in blood pressure.

51. The method of relaxing lower urinary tract tissue of claim 42, wherein the compound is administered in combination with finasteride. SUBSTITUTE SHEET (RULE 26)