U.S. patent number RE37,721 [Application Number 09/594,996] was granted by the patent office on 2002-05-28 for hydroxy-substituted azetidinone compounds useful as hypocholesterolemic agents.
This patent grant is currently assigned to Schering Corporation. Invention is credited to Duane A. Burnett, John W. Clader, Sundeep Dugar, Brian A. McKittrick, Stuart B. Rosenblum.
United States Patent |
RE37,721 |
Rosenblum , et al. |
May 28, 2002 |
**Please see images for:
( Certificate of Correction ) ** |
Hydroxy-substituted azetidinone compounds useful as
hypocholesterolemic agents
Abstract
Hydroxy-substituted azetidinone hypocholesterolemic agents of
the formula ##STR1## or a pharmaceutically acceptable salt thereof,
wherein: Ar.sup.1 and Ar.sup.2 are aryl or R.sup.4 -substituted
aryl; Ar.sup.3 is aryl or R.sup.5 -substituted aryl; X, Y and Z are
--CH.sub.2 --, --CH(lower alkyl)-- or --C(dilower alkyl)--; R and
R.sup.2 are --OR.sup.6, --O(CO)R.sup.6, --O(CO)OR.sup.9 or
--O(CO)NR.sup.6 R.sup.7 ; R.sup.1 and R.sup.3 are H or lower alkyl;
q is 0 or 1; r is 0 or 1; m, n and p are 0-4; provided that at
least one of q and r is 1, and the sum of m, n, p, q and r is 1-6;
and provided that when p is O and r is 1, the sum of m, q and n is
1-5; R.sup.4 is selected from lower alkyl, R.sub.5, --CF.sub.3,
--CN, --NO.sub.2 and halogen R.sup.5 is selected from --OR.sup.6,
--O(CO)R.sup.6, --O(CO)OR.sup.9, --O(CH.sub.2).sub.1-5 OR.sup.6,
--O(CO)NR.sup.6 R.sup.7, --NR.sub.6 R.sup.7, --NR.sup.6
(CO)R.sup.7, --NR.sup.6 (CO)OR.sup.9, --NR.sup.6 (CO)NR.sup.7
R.sup.8, --NR.sup.6 SO.sub.2 R.sup.9, --COOR.sup.6, --CONR.sup.6
R.sup.7, --COR.sup.6, --SO.sub.2 NR.sup.6 R.sup.7, S(O).sub.0-2
R.sup.9, --O(CH.sub.2).sub.1-10 --COOR.sup.6,
--O(CH.sub.2).sub.1-10 CONR.sup.6 R.sup.7, --(lower
alkylene)COOR.sup.6 and --CH.dbd.CH--COOR.sup.6 ; R.sup.6, R.sup.7
and R.sup.8 are H, lower alkyl or aryl-substituted Ic R.sup.9 is
lower alkyl, aryl or aryl-substituted lower alkyl; are disclosed,
as well as a method of lowering serum cholesterol by administering
said compounds, alone or in combination with a cholesterol
biosynthesis inhibitor, pharmaceutical compositions containing
them; and a process for preparing them.
Inventors: |
Rosenblum; Stuart B. (West
Orange, NJ), Dugar; Sundeep (Bridgewater, NJ), Burnett;
Duane A. (Fanwood, NJ), Clader; John W. (Cranford,
NJ), McKittrick; Brian A. (Bloomfield, NJ) |
Assignee: |
Schering Corporation
(Kenilworth, NJ)
|
Family
ID: |
26799377 |
Appl.
No.: |
09/594,996 |
Filed: |
June 15, 2000 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
|
257593 |
Jun 9, 1994 |
5631365 |
|
|
|
102440 |
Sep 21, 1993 |
|
|
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Reissue of: |
617751 |
Mar 18, 1996 |
05767115 |
Jun 16, 1998 |
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Current U.S.
Class: |
514/210.02;
540/200 |
Current CPC
Class: |
A61K
31/35 (20130101); A61P 7/00 (20180101); A61P
9/10 (20180101); A61K 31/40 (20130101); A61P
3/06 (20180101); C07D 205/08 (20130101); A61K
31/395 (20130101); A61K 45/06 (20130101); A61K
9/2018 (20130101); A61K 31/21 (20130101); A61K
9/4858 (20130101); A61K 31/21 (20130101); A61K
2300/00 (20130101); A61K 31/35 (20130101); A61K
2300/00 (20130101); A61K 31/395 (20130101); A61K
2300/00 (20130101); A61K 31/40 (20130101); A61K
2300/00 (20130101); Y02P 20/55 (20151101); Y10S
514/824 (20130101) |
Current International
Class: |
A61P
9/10 (20060101); A61P 9/00 (20060101); A61P
3/00 (20060101); A61P 3/06 (20060101); C07D
205/08 (20060101); C07D 205/00 (20060101); A61K
31/40 (20060101); A61K 31/395 (20060101); A61K
31/35 (20060101); A61K 31/21 (20060101); C07D
205/08 (); A61K 031/395 (); A61P 009/10 (); A61P
003/06 () |
Field of
Search: |
;514/210 ;540/200 |
References Cited
[Referenced By]
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WO |
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WO |
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|
Primary Examiner: Berch; Mark L.
Attorney, Agent or Firm: Komson; Richard C. Auth; Dorothy R.
Morgan & Finnegan, L.L.P.
Parent Case Text
The present application is the United States national application
corresponding to International Application No. PCT/US94/10099,
filed Sep. 14, 1994 and designating the United States, which PCT
application is in turn a continuation-in-part of U.S. application
Ser. No. 08/257593, filed Jun. 9, 1994, U.S. Pat. No. 5,631,365,
which is a continuation-in-part of U.S. application Ser. No.
08/102,440, filed Sep. 21, 1993, abandoned.
Claims
We claim:
1. A compound represented by the formula ##STR50##
or a pharmaceutically acceptable salt thereof, wherein:
Ar.sup.1 and Ar.sup.2 are independently selected from the group
consisting of aryl and R.sup.4 -substituted aryl;
Ar.sup.3 is aryl or R.sup.5 -substituted aryl;
X, Y and Z are independently selected from the group consisting of
--CH.sub.2 --, --CH(lower alkyl)- and --C(dilower alkyl)-;
R and R.sup.2 are independently selected from the group consisting
of --OR.sup.6, --O(CO)R.sup.6, --O(CO)OR.sup.9 and --O(CO)NR.sup.6
R.sup.7 ;
R.sup.1 and R.sup.3 are independently selected from the group
consisting of hydrogen, lower alkyl and aryl;
q is 0 or 1; r is 0 or 1; m, n and p are independently 0, 1, 2, 3
or 4; provided that at least one of q and r is 1, and the sum of m,
n, p, q and r is 2, 3, 4, 5 or 6; and provided that when p is 0 and
r is 1, the sum of m, q and n is 1, 2, 3, 4 or 5;
R.sup.4 is 1-5 substituents independently selected from consisting
of lower alkyl, --OR.sup.6, --O(CO)R.sup.6, --O(CO) OR.sup.9,
--O(CH.sub.2).sub.1-5 OR.sup.6, --(CO)NR.sup.6 R.sup.7, --NR.sup.6
R.sup.7, --NR.sup.6 (CO)R.sup.7, --NR.sup.6 (CO)OR.sup.9,
--NR.sup.6 (CO)NR.sup.7 R.sup.8, --NR.sup.6 SO.sub.2 R.sup.9,
--COOR.sup.6, --CONR.sup.6 R.sup.7, -COR.sup.6, -SO.sub.2 NR.sup.6
R.sup.7, S(O).sub.0-2 R.sup.9, --O(CH.sub.2).sub.1-10 --COOR.sup.6,
--O(CH.sub.2).sub.1-10 CONR.sup.6 R.sup.7, -(lower
alkylene)COOR.sup.6, --CH.dbd.CH--COOR.sup.6, --CF.sub.3, --CN,
--NO.sub.2 and halogen;
R.sup.5 is 1-5 substituents independently selected from the group
consisting of --OR.sup.6, --O(CO)R.sup.6, --O(CO) OR.sup.9,
--O(CH.sub.2).sub.1-5 OR.sup.6, --(CO)NR.sup.6 R.sup.7, --NR.sup.6
R.sup.7, --NR.sup.6 (CO)R.sup.7, --NR.sup.6 (CO)OR.sup.9,
--NR.sup.6 (CO)NR.sup.7 R.sup.8, --NR.sup.6 SO.sub.2 R.sup.9,
--COOR.sup.6, --CONR.sup.6 R.sup.7, --COR.sup.6, --SO.sub.2
NR.sup.6 R.sup.7, S(O).sub.0-2 R.sup.9, --O(CH.sub.2).sub.1-10
--COOR.sup.6, --O(CH.sub.2).sub.1-10 CONR.sup.6 R.sup.7, -(lower
alkylene)COOR.sup.6 and --CH.dbd.CH--COOR.sup.6 ;
R.sup.6, R.sup.7 and R.sup.8 are independently selected from the
group consisting of hydrogen, lower alkyl, aryl and
aryl-substituted lower alkyl; and
R.sup.9 is lower alkyl, aryl or aryl-substituted lower alkyl.
2. A compound of claim 1 wherein Ar.sup.1 is phenyl or R.sup.4
-substituted phenyl, Ar.sup.2 is phenyl or R.sup.4 -substituted
phenyl and Ar.sup.3 is R.sup.5 -substituted phenyl.
3. A compound of claim 2 wherein Ar.sup.1 is R.sup.4 -substituted
phenyl wherein R.sup.4 is halogen; Ar.sup.2 is R.sup.4 -substituted
phenyl wherein R.sup.4 is halogen or --OR.sup.6, wherein R.sup.6 is
lower alkyl or hydrogen; and Ar.sup.3 R.sup.5 -substituted phenyl,
wherein R.sup.5 is --OR.sup.6, wherein R6 is lower alkyl or
hydrogen.
4. A compound of claim 1 wherein X, Y, and Z are each --CH.sub.2
--; R.sup.1 and R.sup.3 are each hydrogen; R and R.sup.2 are each
--OR.sup.6, wherein R.sup.6 is hydrogen; and the sum of m, n, p, q
and r is 2, 3 or 4.
5. A compound of claim 1 wherein m, n and r are each zero, q is 1
and p is 2.
6. A compound of claim 1 wherein p, q and n are each zero, r is 1
and m is 2 or 3.
7. A compound selected from the group consisting of
rel
3(R)-(2(R)-hydroxy-2-phenylethyl)-4(R)-(4-methoxyphenyl)-1-phenyl-2-azetid
inone;
rel
3(R)-(2(R)-hydroxy-2-phenylethyl)-4(S)-(4-methoxyphenyl)-1-phenyl-2-azetid
inone;
3(S)-(1(S)-hydroxy-3-phenylpropyl)-4(S)-(4-methoxyphenyl)-1-phenyl-2-azetid
inone;
3(S)-(1(R)-hydroxy-3-phenylpropyl)-4(S)-(4-methoxyphenyl)-1-phenyl-2-azetid
inone;
3(R)-(1(R)-hydroxy-3-phenylpropyl)-4(S)-(4-methoxyphenyl)-1-phenyl-2-azetid
inone;
rel-3(R)-[(S)-hydroxy-(2-naphthalenyl)methyl]-4(S)-(4-methoxyphenyl)-1-phen
yl)-1-phenyl-2-azetidinone;
rel-3(R)-[(R)-hydroxy-(2-naphthalenyl)methyl]-4(S)-(4-methoxyphenyl)-1-phen
yl-2-azetidinone;
3(R)-(3(R)-hydroxy-3-phenylpropyl)-1,4(S)-bis-(4-methoxyphenyl-2-azetidinon
e;
3(R)-(3(S)-hydroxy-3-phenylpropyl)-1,4(S)-bis-(4-methoxyphenyl-2-azetidinon
e;
4(S)-(4-hydroxyphenyl)-3(R)-(3(R)-hydroxy-3-phenylpropyl-1-(4-methoxyphenyl
)-2-azetidinone;
4(S)-(4-hydroxyphenyl)-3(R)-(3(S)-hydroxy-3-phenylpropyl-1-(4-methoxyphenyl
)-2-azetidinone;
rel
3(R)-[3(RS)-hydroxy-3-[4-methoxymethoxy)-phenyl]propyl]-1,4(S)-bis-(4-meth
oxyphenyl)-2-azetidinone;
1-(4-fluorophenyl)-3(R)-[3(S)-(4-fluorophenyl)-3-hydroxypropyl)]-4(S)-(4-hy
droxyphenyl)-2-azetidinone;
1-(4-fluorophenyl)-3(R)-[3(R)-(4-fluorophenyl)-3-hydroxypropyl)]-4(S)-(4-hy
droxyphenyl)-2-azetidinone;
4(S)-[4-(acetyloxy)phenyl]-3(R)-(3(R)-hydroxy-3-phenylpropyl)-1-(4-methoxyp
henyl)-2-azetidinone;
4(S)-[4-(acetyloxy)phenyl]-3(R)-(3(S)-hydroxy-3-phenylpropyl)-1-(4-methoxyp
henyl)-2-azetidinone;
1-(4-fluorophenyl)-3(R)-[3(S)-(4-fluorophenyl)-3-hydroxypropyl)]-4(S)-[4-(p
henylmethoxy)phenyl]-2-azetidinone;
3(R)-[3(R)-acetyloxy)-3-phenylpropyl]-1,4(S)-bis-(4-methoxyphenyl)-2-azetid
inone;
3(R)-[3(S)-acetyloxy)-3-phenylpropyl]-1,4(S)-bis-(4-methoxyphenyl)-2-azetid
inone;
3(R)-[3(R)-(acetyloxy)-3-(4-fluorophenyl)propyl]-4(S)-[4-(acetyloxy)phenyl]
-1-(4-fluorophenyl)-2-azetidinone;
3(R)-[3(S)-(acetyloxy)-3-(4-fluorophenyl)propyl]-4(S)-[4-(acetyloxy)phenyl]
-1-(4-fluorophenyl)-2-azetidinone;
3(R)-[3(R)-(acetyloxy)-3-(4-chlorophenyl)propyl]-4(S)-[4-(acetyloxy)phenyl]
-1-(4-chlorophenyl)-2-azetidinone;
3(R)-[3(S)-(acetyloxy)-3-(4-chlorophenyl)propyl]-4(S)-[4-(acetyloxy)phenyl]
-1-(4-chlorophenyl)-2-azetidinone; and
rel
1-(4-fluorophenyl)-4(S)-(4-hydroxyphenyl)-3(1R)-(1(R)-hydroxy-3-phenylprop
yl)-2-azetidinone.
8. A pharmaceutical composition for the treatment or prevention of
.[.athersclerosis.]. , .Iadd.atherosclerosis .Iaddend.or for the
reduction of plasma cholesterol levels, comprising an effective
amount of a compound of claim 1 in a pharmaceutically acceptable
carrier.
9. A method of treating or preventing atherosclerosis or reducing
plasma cholesterol levels comprising administering to a mammal in
need of such treatment an effective amount of a compound of claim
1..Iadd.
10. A compound comprising
1-(4-fluorophenyl)-3(R)-[3(S)-(4-fluorophenyl)-3-hydroxypropyl)]-4(S)-4-(h
ydroxyphenyl)-2-azetidinone or a pharmaceutically acceptable salt
thereof..Iaddend..Iadd.
11. A compound represented by the formula: ##STR51##
.Iaddend..Iadd.
12. A pharmaceutical composition for the treatment or prevention of
atherosclerosis, or for the reduction of plasma cholesterol levels,
comprising an effective amount of a compound according to claims 10
or 11 in a pharmaceutically acceptable carrier..Iaddend..Iadd.
13. A method of treating or preventing atherosclerosis or reducing
plasma cholesterol levels comprising administering to a mammal in
need of such treatment an effective amount of a compound according
to claims 10 or 11..Iaddend.
Description
BACKGROUND OF THE INVENTION
The present invention relates to hydroxy-substituted azetidinones
useful as hypocholesterolemic agents in the treatment prevention of
atherosclerosis, and to the combination of a hydroxy-substituted
azetidinone of this invention and a cholesterol bioxynthesis
inhibitor for the treatment and prevention of atherosclerosis. The
invention also relates to a process for preparing
hydroxy-substituted azetidinones.
Atherosclerotic coronary heart disease (CHD) represents the major
cause for death and cardiovascular morbidity in the western world.
Risk factors for atherosclerotic coronary heart disease include
hypertension, diabetes mellitus, family history, male gender, cigar
smoke and serum cholesterol. A total cholesterol level in excess of
225-250 mg/dl is associated with significant elevation of risk of
CHD.
Cholesteryl esters are a major component of atherosclerotic lesions
and the major storage form of cholesterol in arterial wall cells.
Formation of cholesteryl esters is also a key step in the
intestinal absorption of dietary cholesterol. Thus, inhibition of
cholesteryl ester formation and reduction of serum cholesterol is
likely to inhibit the progression of atherosclerotic lesion
formation, decrease the accumulation of cholesteryl esters in the
arterial wall, and block the intestinal absorption of dietary
cholesterol.
A few azetidinones have been reported as being useful lowering
cholesterol and/or in inhibiting the formation of
cholesterol-containing lesions in mammalian arterial walls. U.S.
Pat. No. 4,983,597 discloses N-sulfonyl-2-azetidinones as
anticholesterolemic agents and Ram, et al., in Indian J. Chem.,
Sect. B. 29B, 12 (1990), p. 1134-7, disclose ethyl
4-(2-oxoazetidin-4-yl)phenoxy-alkanoates as hypolipidemic agents.
European Patent Publication 264,231 discloses
1-substituted-4-phenyl-3-(2-oxo-alkylidene)-2-azetidinones as blood
platelet aggregation inhibitors. European Patent 199,630 and
European Patent Application 337,549 disclose elastase inhibitory
substituted azetidinones said to be useful treating inflammatory
conditions resulting in tissue destruction which are associated
with various disease states, e.g. atherosclerosis.
WO93/102048, published Feb. 4, 1993, discloses substituted
.beta.-lactams useful as hypocholesterolemic agents.
The regulation of whole-body cholesterol homeostasis in humans and
animals involves the regulation of dietary cholesterol and
modulation of cholesterol biosynthesis, bile acid biosynthesis and
the catabolism of the cholesterol-containing plasma lipoproteinis.
The liver is the major organ responsible for cholesterol
biosynthesis and catabolism and for this reason, it is a prime
determinant of plasma cholesterol levels. The liver is the site of
synthesis and secretion of very low density lipoproteins (VLDL)
which are subsequently metabolized to low density lipoproteins
(LDL) in the circulation. LDL are the predominant
cholesterol-carrying lipoproteins in the plasma and an increase in
their concentration is correlated with increased
atherosclerosis.
When intestinal cholesterol absorption is reduced, by whatever
means, less cholesterol is delivered to the liver. The consequence
of this action is decreased hepatic lipoprotein (VLDL), production
and an increase in the hepatic clearance of plasma cholesterol,
mostly as LDL. Thus, the net effect of inhibiting intestinal
cholesterol absorption is a decrease in plasma cholesterol
levels.
The inhibition of cholesterol biosynthesis by
3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase
(EC1.1.1.34) inhibitors has been shown to be an effective way to
reduce plasma cholesterol (Witzum, Circulation, 80, 5 (1989), p.
1101-1114) and reduce atherosclerosis. Combination therapy of an
HMG CoA reductase inhibitor and a bile acid sequestrant has been
demonstrated to be more effective in human hyperlipidemic patients
than either agent in monotherapy (Illingworth, Drugs, 36 (Suppl. 3)
(1988), p. 63-71).
SUMMARY OF THE INVENTION
Novel hypocholesterolemic compounds of the present invention are
represented by the formula I ##STR2##
or a pharmaceutically acceptable salt thereof, wherein:
Ar.sup.1 and Ar.sup.2 are independently selected from the group
consisting of aryl and R.sup.4 -substituted aryl;
Ar.sup.3 is aryl or R.sup.5 -substituted aryl;
X, Y and Z are independently selected from the group consisting of
--CH.sub.2 --, --CH(lower alkyl)-- and --C(dilower alkyl)--;
R and R.sup.2 are independently selected from the group consisting
of --OR.sup.6, --O(CO)R.sup.6, --O(CO)OR.sup.9 and --O(CO)NR.sup.6
R.sup.7 ;
R.sup.1 and R.sup.3 are independently selected from the group
consisting of hydrogen, lower alkyl and aryl;
q is 0 or 1; r is 0 or 1; m, n and p are independently 0, 1, 2, 3
or 4; provided that at least one of q and r is 1, and the sum of m,
n, p, q are r is 1, 2, 3, 4, 5 or 6; and provided that when p is 0
and r is 1, the sum of m, q and n is 1, 2, 3, 4, or 5;
R.sup.4 is 1-5 substituents independently selected from the group
consisting of lower alkyl, --OR.sup.6, --O(CO).[.R.sub.6,.].
.Iadd.R.sup.6, .Iaddend.--O(CO)OR.sup.9, --O(CH.sub.2).sub.1-5
OR.sup.6, --O(CO)NR.sup.6 R.sup.7, --NR.sup.6 R.sup.7, --NR.sup.6
(CO)R.sup.7, --NR.sup.6 (CO)OR.sup.9, --NR.sup.6 (CO) NR.sup.7
R.sup.8, --NR.sup.6 SO.sub.2 R.sup.9, --COOR.sup.6, --CONR.sup.6
R.sup.7, --COR.sup.6, --SO.sub.2 NR.sup.6 R.sup.7, S(O).sub.0-2
R.sup.9, --O(CH.sub.2).sub.1-10 --COOR.sup.6,
--O(CH.sub.2).sub.1-10 CONR.sup.6 R.sup.7, --(lower
alkylene)COOR.sup.6, --CH.dbd.CH--COOR.sup.6, --CF.sub.3, --CN,
--NO.sub.2 and halogen;
R.sup.5 is 1-5 substituents independently selected from the group
consisting of --OR.sup.6, --O(CO)R.sup.6, --O(CO)OR.sup.9,
--O(CH.sub.2).sub.1-5 OR.sup.6, --O(CO)NR.sup.6 R.sup.7, --NR.sup.6
R.sup.7, --NR.sup.6 (CO) R.sup.7, --NR.sup.6 (CO)OR.sup.9,
--NR.sup.6 (CO)NR.sup.7 R.sup.8, --NR.sup.6 SO.sub.2 R.sup.9,
--COOR.sup.6, --CONR.sup.6 R.sup.7, --COR.sup.6, --SO.sub.2
NR.sup.6 R.sup.7, S(O).sub.0-2 R.sup.9, --O(CH.sub.2).sub.1-10
--COOR.sup.6, --O(CH.sub.2).sub.1-10 CONR.sup.6 R.sup.7, --(lower
alkylene)COOR.sup.6 and --CH.dbd.CH--COOR.sup.6 ;
R.sup.6, R.sup.7 and R.sup.8 are independently selected from the
group consisting of hydrogen, lower alkyl, aryl and
aryl-substituted lower alkyl; and
R.sup.9 is lower alkyl, aryl or aryl-substituted lower alkyl.
R.sup.4 is preferably 1-3 independently selected substituents, and
R.sup.5 is preferably 1-3 independently selected substituents.
Preferred are compounds of formula I wherein Ar.sup.1 is phenyl or
R.sup.4 -substituted phenyl, especially (4-R.sup.4)-substituted
phenyl, Ar.sup.2 is preferably phenyl or R.sup.4 -substituted
phenyl, especially (4-R.sup.4)-substituted phenyl. Ar.sup.3 is
preferably R.sup.5 -substituted phenyl, especially
(4-R.sup.5)-substituted phenyl. When Ar.sup.1 is
(4-R.sup.4)-substituted phenyl, R.sup.4 is preferably a halogen.
When Ar.sup.2 and Ar.sup.3 are R.sup.4 - and R.sup.5 -substituted
phenyl, respectively, R.sup.4 is preferably halogen or --OR.sup.6
and R.sup.5 is preferably --OR.sup.6, wherein R.sub.6 is lower
alkyl or hydrogen. Especially preferred are compounds wherein each
of Ar.sup.1 and Ar.sup.2 is 4-fluorophenyl and Ar.sup.3 is
4-hydroxyphenyl or 4-methoxyphenyl.
X, Y and Z are each preferably --CH.sub.2 --.R.sup.1 and R.sup.3
are each preferably hydrogen. R and R.sup.2 are preferably
--OR.sup.6 wherein R.sup.6 is hydrogen, or a group readily
metabolizable to a hydroxyl (such as --O(CO)R.sup.6,
.[.--O(CO)OR.sup.9 and.]. .Iadd.--OR.sup.6, especially
.Iaddend.--O(CO)NR.sup.6 R.sup.7, defined above).
The sum of m, n, p, q and r is preferably 2, 3 or 4, more
preferably 3. Preferred are compounds wherein m, n and r are each
zero, q is 1 and p is 2. Also preferred are compounds wherein p, q
and n are each zero, r is 1 and m is 2 or 3. More preferred are
compounds wherein m, n and r are each zero, q is 1, p is 2, Z is
--CH.sub.2 and R is --OR.sup.6 OR.sub.6, especially when R.sup.6 is
hydrogen. Also more preferred are compounds wherein p, q and n are
each zero, r is 1, m is 2, X is --CH.sub.2 -- and R.sup.2 is
--OR.sup.6, especially when R.sup.6 is hydrogen.
Another group of preferred compounds is that wherein Ar.sup.1 is
phenyl or R.sup.4 -substituted phenyl, Ar.sup.2 is phenyl or
R.sup.4 -substituted phenyl and Ar.sup.3 is R.sup.5 -substituted
phenyl. Also preferred are compounds wherein Ar.sup.1 is phenyl or
R.sup.4 -substituted phenyl, Ar.sup.2 is phenyl or R.sup.4
-substituted phenyl, Ar.sup.3 is R.sup.5 -substituted phenyl, and
the sum of m, n, p, q and r is 2, 3 or 4, more especially 3. More
preferred are compounds wherein Ar.sup.1 is phenyl or R.sup.4
-substituted phenyl, Ar.sup.2 is phenyl or R.sup.4 -substituted
phenyl Ar.sup.3 is R.sup.5 -substituted phenyl, and wherein m, n
and r are each zero, q is 1 and p is 2, or wherein p, q and n are
each zero, r is 1 and m is 2 or 3.
This invention also relates to a method of lowering the serum
cholesterol level in a mammal in need of such treatment comprising
administering an effective amount of a compound of formula I. That
is, the use of a compound of the present invention as an
hypocholesterolemic agent is also claimed.
In still another aspect, the present invention relates to a
pharmaceutical composition comprising a serum cholesterol-lowering
effective amount of a compound of formula I in a pharmaceutically
acceptable carrier.
The present invention also relates to a method of reducing plasma
cholesterol levels, and to a method of treating or preventing
atherosclerosis, comprising administering to a mammal in need of
such treatment an effective amount of a combination of a
hydroxy-substituted azetidinone cholesterol absorption inhibitor of
formula I and a cholesterol biosynthesis inhibitor. That is, the
present invention relates to the use of a hydroxy-substituted
azetidinone cholesterol absorption inhibitor of formula I for
combined use with a cholesterol biosynthesis inhibitor (and,
similarly, use of a cholesterol biosynthesis inhibitor for combined
use with a hydroxy-substituted azetidinone cholesterol absorption
inhibitor of formula I) to treat or prevent atherosclerosis or to
reduce plasma cholesterol levels.
In yet another aspect, the invention relates to a pharmaceutical
composition comprising an effective amount of a hydroxy-substituted
azetidinone cholesterol absorption inhibitor of formula I, a
cholesterol biosynthesis inhibitor, and a pharmaceutically
acceptable carrier. In a final aspect, the invention relates to a
kit comprising in one container an effective amount of a
hydroxy-substituted azetidinone cholesterol absorption inhibitor of
formula I in a pharmaceutically acceptable carrier, and in a
separate container, an effective amount of a cholesterol
biosynthesis inhibitor in a pharmaceutically acceptable
carrier.
In yet another aspect, the invention relates to a process for
preparing certain compounds of formula I comprising the steps:
(a) treating with a strong base a lactone of the formula
##STR3##
wherein R' and R.sup.2 ' are R and R.sup.2, respectively, or are
suitably protected hydroxy groups; Ar.sup.10 is Ar.sup.1, a
suitably protected hydroxy substituted aryl or a suitably protected
amino-substituted aryl; and the remaining variables are as defined
above, provided that in lactone of formula B when n and r are each
zero, p is 1-4;
(b) reacting the product of step (a) with an imine of the formula
##STR4##
wherein Ar.sup.20 is Ar.sup.2, a suitably protected
hydroxy-substituted aryl or a suitably protected amino-substituted
aryl; and Ar.sup.30 is Ar.sup.3, a suitably protected
hydroxy-substituted aryl or a suitably protected amino-substituted
aryl;
c) quenching the reaction with an acid;
d) optionally removing the protecting groups from R', R.sup.2',
Ar.sup.10, Ar.sup.20 and Ar.sup.30, when present; and
e) optionally functionalizing hydroxy or amino substituents at R,
R.sup.2, Ar.sup.1, Ar.sup.2 and Ar.sup.3.
Using the lactones shown above, compounds of formula IA and IB are
obtained as follows: ##STR5##
wherein the variables are as defined above; and ##STR6##
wherein the variables are as defined above.
DETAILED DESCRIPTION
As used herein, the term "lower alkyl" means straight or branched
alkyl chains of 1 to 6 carbon atoms.
"Aryl" means phenyl, naphthyl, indenyl, tetrahydronaphthyl or
indanyl.
"Halogen" refers to fluorine, chlorine, bromine or iodine
atoms.
The above statement, wherein R.sup.6, R.sup.7 and R.sup.8 are said
to be independently selected from a group of substituents, means
that R.sup.6, R.sup.7 and R.sup.8 are independently selected, but
also that where an .[.R.sub.6.]. , .Iadd.R.sup.6 .Iaddend.or
R.sup.8 variable occurs more than once in a molecule, those
occurrences are independently selected (e.g., if R is --OR.sup.6
wherein R.sup.6 is hydrogen, R.sup.4 can be --OR.sup.6 wherein
R.sup.6 is lower alkyl).
Compounds of the invention have at least one asymmetric carbon atom
and therefore all isomers, including enantiomers and diastereomers
are contemplated as being part of this invention. The invention
includes d and .[.I.]. .Iadd.l .Iaddend.isomers in both pure form
and in admixture including racemic mixtures. Isomers can be
prepared using conventional techniques, either by reacting chiral
starting materials or by separating isomers of a compound of
formula I. Isomers may also include geometric isomers, e.g. when a
double bond is present. All such geometric isomers are contemplated
for this invention.
Those skilled in the art will appreciate that for some compounds of
formula I, one isomer will show greater pharmacological activity
than another isomer.
Compounds of the invention with an amino group can form
pharmaceutically acceptable salts with organic and inorganic acids.
Examples of suitable acids for salt formation are hydrochloric,
sulfuric, phosphoric, acetic, citric, oxalic, malonic, salicylic,
malic, fumaric, succinic, ascorbic, maleic, methanesulfonic and
other mineral and carboxylic acids well known to those in the art.
The salt is prepared by contacting the free base form with a
sufficient amount of the desired acid to produce a salt. The free
base form may be regenerated by treating the salt with a suitable
dilute aqueous base solution such as dilute aqueous sodium
bicarbonate. The free base form differs from its respective salt
form somewhat in certain physical properties, such as solubility in
polar solvents, but the salt is otherwise equivalent to its
respective free base form for purposes of the invention.
Certain compounds of the invention are acidic (e.g., those
compounds which possess a carboxyl group). These compounds form
pharmaceutically acceptable salts with inorganic and organic bases.
Examples of such salts are the sodium, potassium, calcium,
aluminum, gold and silver salts. Also included are salts formed
with pharmaceutically acceptable amines such as ammonia, alkyl
amines, hydroxyalkylamines, N-methylglucamine and the like.
Cholesterol biosynthesis inhibitors for use in the combination of
the present invention include HMG CoA reductase inhibitors such as
lovastatin, pravastatin, fluvastatin, simvastatin, and Cl-981; HMG
CoA synthetase inhibitors, for example L-659,699
((E,E)-11-[3'-R-(hydroxy-methyl)-4'-oxo-2'-R-oxetanyl]-3,5,7R-trimethyl-2,
4-undecadienoic acid); squalene synthesis inhibitors, for example
squalestatin 1; and squalene epoxidase inhibitors, for example,
NB-598
((E)-N-ethyl-N-(6,6-dimethyl-2-hepten-4-ynyl)-3-[(3,3'-bithiophen-5-yl)met
hoxy]benzene-methanamine hydrochloride) and other cholesterol
biosynthesis inhibitors such as DMP-565. Preferred HMG CoA
reductase inhibitors are lovastatin, pravastatin and
simvastatin.
Compounds of formula I can be prepared by known methods, for
example those described below and in WO93/02048. ##STR7##
Compounds of formula Ia and Ib, wherein Ar.sup.1, Ar.sup.2,
.[.Ar3,.]. .Iadd.Ar.sup.3 .Iaddend.X, Y, Z, R.sup.1, R.sup.2,
R.sup.3, m, n, p, q and r are as defined above, can be prepared by
treatment of an ester of formula III, wherein R.sup.10 is lower
alkyl such as ethyl or a chiral moiety such as menthyl or
10-(diisopropylsulfonamido)isobornyl, and the remaining variables
are as defined above, with a strong base such as lithium
diisopropylamide (LDA) in a suitable solvent such as
tetrrahydrolithium (THF) at -78.degree. C. A solubilizing agent
such as hexamethylphosphoric triamide (HMPA) may optionally be
added as a cosolvent. An imine of formula 11, wherein Ar.sup.20 and
Ar.sup.30 are as defined above, is added, the reaction mixture is
either warmed to room temperature or maintained at a suitable low
temperature such as -78.degree. C. for the appropriate time,
followed by quenching with a suitable acid such as 1N HCl. The
product is isolated using conventional purification techniques.
When a protecting group as defined in Table 1 (below) is present on
one or more of the optionally protected groups, an additional step
comprising removal of the protecting group by conventional
techniques is needed. However, for compounds of formula Ia, Ib, or
any compound of formula I wherein a protected hydroxy group
Ar.sup.10, Ar.sup.20, Ar.sup.30, R' or R.sup.2 ' is an alkoxy or
benzyloxy group, such a protecting group need not be removed to
obtain a compound of formula I. When a chiral ester of formula III
is used, the resulting compound of formula Ia or Ib is not
racemic.
Imines of formula II (Ar.sup.30 --CH.dbd.N--Ar.sup.20) can be
prepared from aldehydes of the formula Ar.sup.30 --CHO and amines
of the formula .[.Ar.sup.+ --CHO and.]. .Iadd.Ar.sup.20 --NH.sub.2
.Iaddend.by procedures well known in the art. Aldehydes of formula
.[.Ar.sup.+.]. .Iadd.Ar.sup.30.Iaddend.--CHO and amines of formula
Ar.sup.20 --NH.sub.2 are commercially available or can be prepared
via known procedures. ##STR8##
Compounds of formula Ic and Id, wherein the variables are as
defined above, can be prepared by a process comprising the
following steps:
(a) Treat a lactone of formula IV, wherein the variables are as
defined above, with a strong base such as an alkyllithium (e.g.,
n-butyl-lithium), a metal hydride (e.g., sodium hydride), a metal
alkoxide (e.g., sodium methoxide), a metal halide (e.g.,
TiCl.sub.4), metal exchange of the lithium enolate with a metal
halide (e.g., zinc chloride), metal exchange of the lithium enolate
with a metal alkyl (e.g., 9-borabicyclononyl triflate), or,
preferably, a metalamide (e.g., LDA), in a suitable anhydrous
organic solvent such as dry THF, ether or benzene, in a dry, inert
atmosphere, e.g., under nitrogen. The reaction is carried out at
about 0.degree. C. to about -85.degree. C., preferably about
-78.degree. C., over a period of about 5 to about 60 minutes,
preferably about 30 minutes. 1-50% of solubilizing cosolvents may
optionally be added, preferably about 10% HMPA.
(b) Add an imine of formula 11, wherein Ar.sup.20 and Ar.sup.30 are
as defined above, to the product of step (a) over a period of 5 to
60 minutes, preferably 30 minutes, maintaining the reaction mixture
at about 0.degree. C. to about -85.degree. C., preferably about
-78.degree. C., for 1 to 12 hours, preferably about 3 hours, or
warming the reaction mixture over that time period at a rate of
about 10.degree. C. per hour to about 70.degree. C. per hour,
preferably about 30.degree. C. per hour, to a temperature of about
20.degree. C.
(c) Quench the reaction with a suitable acid such as HCl (1N).
(d) The protecting groups on R', R.sup.2 ', Ar.sup.10, Ar.sup.20
and Ar.sup.30, when present, are removed, if desired, by methods
well known in the art, for example silyl protecting groups are
removed by treatment with fluoride.
e) Compounds of formula I wherein any of R and R.sup.2, when
present, are OR.sup.6 wherein R.sup.6 is hydrogen, can be converted
by well known methods to other compounds of formula I wherein R and
R.sup.2 are functionalized, i.e., are independently selected from
the group consisting of OR.sup.6a, --O(CO)R.sup.6, --O(CO)OR.sup.9
and --O(CO)NR.sup.6 R.sup.7, wherein R.sup.6, R.sup.7 and R.sup.9
are as defined above and R.sup.6a is lower alkyl, aryl, or
aryl-lower alkyl. For example, treatment of the alcohol with an
alkyl halide in the presence of a suitable base such as NaH will
afford alkoxy-substituted compounds (i.e., R or R.sup.2 is
OR.sup.6, wherein R.sup.6 is lower alkyl); treatment of the alcohol
with an acylating agent such as acetylchloride will result in
compounds wherein R or R.sup.2 is --OC(O)R.sup.6 ; treatment of the
alcohol with phosgene followed .Iadd.by an .Iaddend.alcohol of the
formula HOR.sup.9 affords compounds substituted with a --OC
(O)OR.sup.9 group; and treatment of the alcohol with phosgene
followed by an amine of the formula HNR.sup.6 R.sup.7 affords
compounds wherein R or R.sup.2 is --OC(O)NR.sup.6 R.sup.7.
Compounds of formula I wherein any Ar.sup.1, Ar.sup.2 or Ar.sup.3
has a hydroxy or amino group can be similarly functionalized to
obtain other compounds of formula 1, i.e., wherein R.sup.4 and
R.sup.5 are independently --OR.sup.6a, --O(CO)R.sup.6,
--O(CO)OR.sup.9, --O(CH.sub.2).sub.1-5 OR.sup.6, --O(CO)NR.sup.6
R.sup.7, --NR.sup.6 R.sup.7, --NR.sup.6 (CO) R.sup.7, --NR.sup.6
(CO)OR.sup.9, --NR.sup.6 (CO)NR.sup.7 R.sup.8 or --NR.sup.6
SO.sub.2 R.sup.9.
The product of step c, d or e is isolated using conventional
purification techniques such as extraction, crystallization or,
preferably, silica gel 60 chromatography. When a chiral lactone is
used, the resulting compound of formula Ic or Id is not
racemic.
Using the procedure described in steps (a)-(e), lactones of formula
IVa can be used to prepare compounds of formula Ig and Ih, provided
that when n and r are each zero, p is 1-4: ##STR9##
Lactones of formulae IV and IVa are known in the art or can be
prepared by methods well known in the art. See, for example, U.S.
Pat. No. 4,375,475 and J. Agric. Food Chem., 30 (5) (1982) p.
920-4. ##STR10##
Azetidinones of formula V, wherein Ar.sup.20 and Ar.sup.30 are as
defined above, can be reacted to form compounds of formula Ie and
If i.e., compounds of formula I wherein r is 1, R.sup.2 is hydroxy,
and p is zero) by treatment of azetidinone V with a strong base
such as lithium .[.iosoproptylcyclohexylamide.].
.Iadd.isopropylcyclohexylamide .Iaddend.in a suitable solvent such
as THF in the presence or .[.absent.]. .Iadd.absence .Iaddend.of
HMPA at -78.degree. C., followed by the addition of an aldehyde or
ketone of VI, wherein Ar.sup.10, X, Y, R', R.sup.1, R.sup.3, m, n
and q are as defined above. As in the case of Method A, protecting
groups at Ar.sup.10, Ar.sup.20, Ar.sup.30, R' and R.sup.2' are
removed as necessary.
This process provides several of the possible diastereomers which
can be separated by a combination of crystallization, silica gel
chromatography and HPLC, using techniques well known in the art.
The remaining diastereomers can be obtained by inversion reactions
such as the Mitsunobu reaction sequence outlined below, wherein
partial structures of formula If are shown: ##STR11##
In the above known process, DEAD is diethylazodicarboxylate and
PPh.sub.3 is triphenylphosphine. The reactants are stirred at room
temperature overnight and the resultant formate ester is converted
to the corresponding hydroxy compound with the desired
stereochemistry. ##STR12##
Compounds of formula Ia as defined above can be prepared by
reacting a chiral auxiliary such as the compound of formula VIII
with an activated carboxylic acid derivative of formula VII, for
example an acid chloride (L.dbd.Cl), a mixed anhydride formed with
phenyl phosphorodichloridate (L.dbd.OP(O)(Cl)OPh), an
N-methyl-pyridinium ester formed from the reaction of an acid with
N-methyl-2-chloropyridinium iodide (L=2-oxy-N-methylpyridinium
iodide), and a 2-thiopyridyl ester formed from the reaction of an
acid chloride and 2-thiopyridine, wherein the remaining variables
are as defined above; enolizing the resultant product, for example
with TiCl.sub.4 and tetramethylethylenediamine (TMEDA); condensing
with an aldehyde, Ar.sup.30 CHO; hydrolyzing to the corresponding
acid, then reacting the compound of formula IX with an amine,
Ar.sup.20 NH.sub.2 ; and cyclizing the resultant compound of
formula X, with, for example a trialkylphosphine and a
dialkylazodicarbo)ylate. As in the case of Method A, protecting
groups at Ar.sup.10, Ar.sup.20, Ar.sup.30, R' and R.sup.2' are
removed as necessary. This procedure is described in detail in
.[.WO93/102048.]. .Iadd.WO93/02048.Iaddend.. ##STR13##
Compounds of formula Ia as defined above can also be prepared
treatment of an imine of formula .[.11,.]. .Iadd.II,
.Iaddend.wherein Ar.sup.20 and Ar.sup.30 are as defined above, with
an activated carboxylic acid derivative of formula VII as defined
above in the presence of a tertiary amine base such as
triethylamine, tributylamine or diethylisopropylamine in an inert
solvent such as CH.sub.2 Cl.sub.2. Again, as in the case of Method
A, protecting groups at Ar.sup.10, Ar.sup.20, Ar.sup.30, R' and
R.sup.2 ' are removed as necessary. Use of other bases, e.g.,
pyridine, favors formation of compounds of formula Ib.
##STR14##
In the first step, compound XII is dissolved in a suitable solvent,
e.g., anhydrous CH.sub.2 Cl.sub.2, and treated with a Lewis acid,
e.g., TiCl.sub.4 at about -60.degree. C. to 0.degree. C.,
preferably at about -25.degree. C., under a dry, inert atmosphere,
e.g., argon. A tertiary amine base such as TMEDA is added and the
mixture stirred at about -60.degree. C. to 0.degree. C., preferably
at about -25.degree. C. to -15.degree. C., for a period of about 1
h. An imine of formula Ar.sup.30 CH.dbd.NAr.sup.20 is added neat or
optionally as a solution in a suitable solvent, e.g. anhydrous
CH.sub.2 Cl.sub.2, over a period of about 5 min, and the reaction
is stirred vigorously at about -60.degree. C. to 0.degree. C.,
preferably at about -25.degree. C. to -15.degree. C., for about 3
to 6 h, preferably about 4 h or until the reaction is complete by
TLC. An acid, e.g. acetic acid, is added to reaction at the
reaction temperature and the mixture is allowed to warm to room
temperature slowly with stirring for about 1-3 hours, preferably
about 2 hours. The compound of formula XII is isolated by
extraction with a suitable solvent, e.g. CH.sub.2 Cl.sub.2, then
purified by crystallization or silica gel chromatography.
In the second step, the product is treated with a strong
non-nucleophilic base, such as sodium or lithium
bistrimethylsilylamide at about -78.degree. C. to 100.degree. C.
After reaction, the mixture is poured into aqueous tartaric acid
and the product isolated from the organic layer. As in the case of
Method A, protecting groups at Ar.sup.10, Ar.sup.20, Ar.sup.30, R'
and R.sup.2' are removed as necessary. This process, including the
preparation of the starting material of formula XII, is also
described in greater detail in WO93/02048. ##STR15##
Compound of formula Ig' and Ih' (i.e., compounds of formula I
wherein R is OH), wherein R.sup.2 ' is a protected hydroxy group as
defined above, and the remaining variables are as defined above,
can be prepared by reacting an imine of formula .[.11.]. .Iadd.II
.Iaddend.and a carboxylic acid derivative of formula XIV, wherein
the variables are as defined above, according to Method D, followed
by oxidation of the resultant halide of formula XV by treatment
with an oxidizing agent such as trimethylamine oxide, CrO.sub.3 or
ozone in a solvent such as DMSO. The resultant aldehyde or ketone
of formula XVI is then reacted with an aryl organometallic reagent
(e.g., Ar.sup.10 X.sub.m MgBr, Ar.sup.10 X.sub.m Li, Ar.sup.10
X.sub.m MgCl or Ar.sup.10 X.sub.m CeCl.sub.2) to obtain a compound
of formula Ig' or Ih'. As described above, the Ar.sup.10,
Ar.sup.20, Ar.sup.30 and R.sup.2' substituents can be converted to
the desired Ar.sup.1, Ar.sup.2, Ar.sup.3 and R.sup.2 substituents
by procedures well known in the art. ##STR16##
Compounds of formula Ii having a hydroxy substituent on the side
chain adjacent to the Ar.sup.1 group (i.e., compounds of formula I
wherein m is 0) can be prepared by heating a compound of formula
XVII, prepared by Method D, above, wherein the variables are as
defined above, for about 1-6 hours at about 60.degree. C. to
100.degree. C. with a halogenating agent such as N-bromosuccinimide
(NBS) in a suitable solvent such as CCl.sub.4 in the presence of an
initiating agent such as benzoyl peroxide. The resultant compound
of formula XVIII, wherein Hal is Cl, Br or I and the remaining
variables are as defined above, is then heated in a suitable
solvent such as CH.sub.2 Cl.sub.2 with a tetraalkyl-ammonium salt
such as tetra n-butyl-ammonium hydroxide (n-Bu.sub.4 NOH) to obtain
the compound of formula Ia. Alternatively, compound XVIII can be
heated in a suitable solvent such as CH.sub.2 Cl.sub.2 with tetra
n-butylammonium trifluoroacetate (n-Bu.sub.4 NOC(O)CF.sub.3)
followed by treatment with a mild base such as ethanol saturated
with .[.NH3.]. .Iadd.NH.sub.3 .Iaddend.to obtain compound Ii,
##STR17##
Compounds of formula Ij (i.e., compounds of formula I wherein R is
OH, R.sup.1 is H and q is 1) are prepared from compound XIX in 2
steps. First, a compound of formula XIX, wherein the variables are
as defined above, is dissolved in a suitable anhydrous solvent,
e.g. THF, at about -20.degree. C. to about 22.degree. C.,
preferably at about 0.degree. C. under a dry inert atmosphere, e.g.
argon and adding a transition metal source, e.g.
tetrakis(triphenylphosphine)-palladium or palladium
acetate/triphenyl phosphine. An organometallic of formula Ar.sup.10
-X.[.m.]. .Iadd..sub.m.Iaddend.-Met, wherein in Ar.sup.10, X and m
are as defined above and Met is, for example, ZnCl or B(OH).sub.2,
is added to the reaction mixture at about -20.degree. C. to about
22.degree. C., preferably at about 0.degree. C., the reaction
mixture is stirred for about 15 min to 4 h, preferably about 1 h,
and is then allowed to warm to about 22.degree. C. Addition of
dilute acid, e.g. 1N HCl, followed by extraction with a suitable
organic solvent, e.g. ethyl acetate (EtOAc), produces compound
XX.
The ketone of-formula XX is dissolved in a suitable .Iadd.solvent
.Iaddend.e.g. CH.sub.3 OH, a hydrogenation catalyst is added, e.g.
Pd on carbon, and the mixture is exposed to H.sub.2 gas under a
pressure of about 14 psi to 100 psi, preferably about 60 psi for
about 1 to 24 h, preferably, about 16 h. The hydrogenation catalyst
is removed by filtration and the solvent is removed in vacuo to
produce a compound Ij as a mixture of alcohol diastereomers which
can be separated by conventional means.
Alternatively, a ketone of formula XX is dissolved in a suitable
solvent, e.g. THF, at about -40.degree. C. to about 22.degree. C.,
preferably at about 0.degree. C., and a suitable reducing agent
such as NaBH.sub.4, a substituted borohydride (e.g., [cbz-proline]
.sub.3 BHNa) or a borane is adlded, optionally in the presence of a
suitable chiral promotor present either in catalytic or
stoichiometric amounts, e.g., chiral borane of structures:
##STR18##
Addition of dilute acid, e.g., 1N HCl, followed by extraction with
a suitable solvent produces compounds of formula Ij. As above,
protecting groups at Ar.sup.10, Ar.sup.20, Ar.sup.30 and R.sup.2 '
are removed as necessary. When either a chiral reagent or a chiral
promotor is used, the resulting product is non-racemic.
Compounds of formula XIX can be prepared by a multi-step procedure
as represented below: ##STR19##
Compounds of formula XXI, wherein R.sup.10 is lower alkyl and the
remaining .Iadd.variables are as defined above, are commercially
available or can be prepared by treating the corresponding
carboxylic acid (i.e., compounds wherein the Cl is replaced by a
hydroxy group) with a chlorinating agent, e.g. SOCl.sub.2 or oxalyl
chloride, under a dry atmosphere, neat or in a suitable inert
organic solvent, e.g. toluene at about 40.degree. C. to 110.degree.
C., preferably about 70.degree. C.; alternatively, a catalyst made
be added, e.g. dimethylformamide (DMF), the reaction is conducted
at about 22.degree. C., and the solvent and excess reagents are
removed in vacuo. The compound XXI is reacted with a chiral
auxiliary such as (S)-4-phenyl-2-oxazolidinone according to the
following procedure; a chiral auxiliary is treated with a strong
base such as an alkyllithium, a metal hydride or a tertiary amine
base such as triethylamine, in a suitable anhydrous organic
solvent, e.g., dry THF, under a dry, inert atmosphere, e.g. argon
at about -85.degree. C., to 22.degree. C., preferably about
0.degree. C., for about 10 min to 60 min, preferably about 30
minutes. The resulting anion is reacted, without isolation, with
compound XXI in a suitable anhydrous organic solvent, e.g. dry THF,
under a dry, inert atmosphere, e.g. argon at about -85.degree. C.
to about 22.degree. C., preferably 0.degree. C., for about 30 min
to 60 min, preferably 30 min. The reaction is warmed to about
22.degree. C. and continued for 1 to 12 h, preferably 6 h. Water is
added and compound XXII is isolated by extraction and purified by
crystallization.
The compound of formula XXII is treated in the same manner as
described in step 1 of Method E to obtain a compound
XXIII..Iaddend.
Azetidinone ring closure can be accomplished by alternative
procedures. By one method, a compound of formula XXIII is treated
with a strong non-nucleophilic base, such as sodium or
lithium-bistrimethylsilylamide, in a suitable inert organic
solvent, e.g. CH.sub.2 Cl.sub.2, at about -78.degree. C. to about
10.degree. C., preferably about 0.degree. C. The mixture is stirred
for about 1 to 2 hours while gradually warming to about 22.degree.
C. Compound XXIV is isolated by conventional extraction with
CH.sub.2 Cl.sub.2. In another, two-step method, a compound of
formula XXIII is first treated with mild silylating agent, e.g.
N,O-bis(trimethylsilyl)acetamide at about 0.degree. C. to about
100.degree. C., preferably about 40.degree. C. for about 10 min to
60 min, preferably 30 min, then treated with a fluoride anion
source, e.g. tetrabutylammonium fluoride (TBAF), at about 0.degree.
C. to about 100.degree. C., preferably 40.degree. C., and allowed
to stir for about 0.5 to about 4 hours, preferably about 2 hours.
Compound XXIV is isolated by conventional extraction methods.
The compound of formula XXIV is hydrolysed by a suitable base, e.g.
LiOH, in a suitable solvent, e.g. 66% CH.sub.3 OH/water at about
0.degree. C. to about 50.degree. C., preferably 22.degree. C., for
about 1 to 4 hours, preferably 2 hours, then extracted with a
suitable solvent, e.g. EtOAc. The resulting acid is converted to
the acid chloride as described above by treatment with a
chlorination agent, e.g. oxalyl chloride, to afford compound
##STR20##
Compounds of formula Ik, wherein Ar.sup.1, Ar.sup.2, Ar.sup.3 and
R.sup.1 are as defined above, one of X" and Y" is --CH.sub.2
CH.sub.2 -- and the other is selected from the group consisting of
--CH.sub.2 CH.sub.2 --, --CH.sub.2 --, --CH(lower alkyl)-, --CH
(dilower alkyl) and a bond, are prepared by oxidation of an alkene
of formula XXV, wherein one of X' and Y' is --CH.dbd.CH-- and the
other is --CH.dbd.CH--, --CH.sub.2 --, --CH.sub.2 CH.sub.2 --,
--CH(lower alkyl)-, --CH(dilower alkyl) or a bond, and the
remaining variables are as defined above, can be prepared by the
following two step procedure.
A compound of formula XXV, which can be prepared by Method D,
above, is treated with an oxidizing agent such as SeO.sub.2,
phenylselenic anhydride or CrO.sub.3 in a suitable solvent such as
dioxane at about 22.degree. to 100.degree. C. for about 0.5 to 12
hours. After the starting material is consumed as determined by
TLC, or 12 hours, the reaction is cooled to about 22.degree. C. and
the product XXVI is isolated by extraction.
In the second step, an allylic alcohol of formula XXVI is dissolved
in a suitable solvent, e.g., EtOAc, a hydrogenation catalyst added,
e.g., Pd on carbon, and the mixture is exposed to H.sub.2 gas under
a pressure of about 14 psi to 60 psi for about 1 to 12 hours. The
hydrogenation catalyst is removed in vacuo to obtain a compound of
formula Ik. ##STR21##
Alcohols of formula Im and In (i.e., compounds of formula I where r
is 1, R.sup.2 is --OH, R.sup.3 is hydrogen and p is 0) can be
selectively obtained from ketones of formula XXVII in three steps
comprising bromination, reduction and debromination. Since the
stereochemistry of the major isomers of alcohols XXIXa and XXIXb
are different, one can selectively prepare either diastereomeric
alcohol.
In the above process, a ketone of formula XXVII, which can be
prepared by oxidation of the corresponding hydroxy compound by well
known methods, is halogenated, for example by treatment in an inert
solvent, e.g., THF, with NaH followed by N-bromosuccinimide, to
obtain a mixture of 3-bromo-ketone compounds XXVIII (a and b).
Compounds .[.15.]. XXVIIIa and XXVIIIb are then separately reduced
to the corresponding alcohols, for example by treatment with
magnesium trifluoroacetate (Mg(TFA).sub.2) and t-butylamine borane
(t-Bu--NH.sub.2 --BH.sub.3) in an inert solvent such as THF at a
temperature of about -78.degree. C. to 0.degree. C. The resultant
alcohols XXIX are dehalogenated by treatment with
tris(trimethylsilyl)silane ((TMS).sub.3 SiH) in a solvent such as
toluene in the presence of a radical initiator such as
2,2'-azobisisobutyronitrile (AIBN) to obtain a mixture of isomers
Im and In which can be separated into individual enantiomers by
conventional means, e.g., HPLC. Again, protecting groups at
Ar.sup.10, Ar.sup.20, Ar.sup.30 and R' are removed as
necessary.
Starting compounds III, V, VI, VII, VIII, XIV, XVII, XXI and XXV
are all either commercially available or well known in the art and
can be prepared via known methods.
Reactive groups not involved in the above processes can be
protected during the reactions with conventional protecting groups
which can be removed by standard procedures after the reaction. The
following Table 1 shows some typical protecting groups:
TABLE 1 Group to be Group to be Protected and Protected Protecting
Group --COOH --COOalkyl, --COObenzyl, --COOphenyl ##STR22##
##STR23## ##STR24## ##STR25## --NH.sub.2 ##STR26## --OH ##STR27##
--OSi(CH.sub.3).sub.3, or --OCH.sub.2 phenyl
We have found that the compounds of this invention lower serum
lipid levels, in particular serum cholesterol levels. Compounds of
this invention have been found to inhibit the intestinal absorption
of cholesterol and to significantly reduce the formation of liver
cholesteryl esters in animal models. Thus, compounds of this
invention are hypocholesterolemic agents by virtue of their ability
to inhibit the intestinal absorption and/or esterification of
cholesterol; they are, therefore, useful in the treatment and
prevention of atherosclerosis in mammals, in particular in
humans.
The in vivo activity of the compounds of formula I can be
determined by the following procedure:
In Vivo Assay of .[.Hypoligidemic.]. .Iadd.Hypolipidemic
.Iaddend.Agents Using the Hyperlipidemic Hamster
Hamsters are separated into groups of six and given a controlled
cholesterol diet (Purina Chow #5001 containing 0.5% cholesterol)
for seven days. Diet consumption is monitored to determine dietary
cholesterol exposure in the face of test compounds. The animals are
dosed with the test compound once daily beginning with the
initiation of diet. Dosing is by oral gavage of 0.2 mL of corn oil
alone (control group) or solution (or suspension) of test compound
in corn oil. All animals moribund or in poor physical condition are
euthanized. After seven days, the animals are anesthetized by
intramuscular (IM) injection of ketamine and sacrificed by
decapitation. Blood is collected into vacutainer tubes containing
EDTA for plasma lipid analysis and the liver excised for tissue
lipid analysis. Lipid analysis is conducted as per published
procedures (Schnitzer-Polokoff, R., et al. Comp. Biochem. Physiol.,
99A, 4 (1991), p. 665-670) and data is reported as percent
reduction of lipid versus control.
The present invention also relates to a pharmaceutical composition
comprising a compound of formula I and a pharmaceutically
acceptable carrier. The compounds of formula I can be administered
in any conventional dosage form, preferably an oral dosage form
such as a capsule, tablet, powder, cachet, suspension or solution.
The formulations and pharmaceutical compositions can be prepared
using conventional pharmaceutically acceptable excipients and
additives and conventional techniques. Such pharmaceutically
acceptable excipients and additives include non-toxic compatible
fillers, binders, disintegrants, buffers, preservatives,
anti-oxidants, lubricants, flavorings, thickeners, coloring agents,
emulsifiers and the like.
The daily hypocholesteremic dose of a compound of formula I is
about 0.1 to about 30 mg/kg of body weight per day, preferably
about 0.1 to about 15 mg/kg. For an average body weight of 70 kg,
the dosage level is therefore from about 5 mg to about 1000 mg of
drug per day, given in a single dose of 2-4 divided doses. The
exact dose, however, is determined by the attending clinician and
is dependent on the potency of the compound administered, the age,
weight, condition and response of the patient.
For the combinations of this invention wherein the hydroxy
substituted azetidinone is administered in combination with a
cholesterol biosynthesis inhibitor, the typical daily dose of the
cholesterol biosynthesis inhibitor is 0.1 to 80 mg/kg of mammalian
weight per day administered in single or divided dosages, usually
once or twice a day; for example, for HMG CoA reductase inhibitors,
about 10 to about 40 mg per dose is given 1 to 2 times a day,
giving a total daily dose of about 10 to 80 mg per day, and for the
other cholesterol biosynthesis inhibitors, about 1 to 1000 mg per
dose is given 1 to 2 times a day, giving a total daily dose of
about 1 mg to about 200 mg per day. The exact dose of any component
of the combination to be administered is determined by the
attending clinician and is dependent on the potency of the compound
administered, the age, weight, condition and response of the
patient.
Where the components of a combination are administered separately,
the number of doses of each component given per day may not
necessarily be the same, e.g. where one component may have a
greater duration of activity, and will therefore need to be
administered less frequently.
Since the present invention relates to the reduction of plasma
cholesterol levels by treatment with a combination of active
ingredients wherein said active ingredients may be administered
separately, the invention also relates to combining separate
pharmaceutical compositions in kit form. That is, a kit is
contemplated wherein two separate units are combined: a cholesterol
biosynthesis inhibitor pharmaceutical composition and a hydroxy
substituted azetidinone cholesterol absorption inhibitor
pharmaceutical composition. The kit will preferably include
directions for the administration of the separate components. The
kit form is particularly advantageous when the separate components
must be administered in different dosage forms (e.g. oral and
parenteral) or are administered at different dosage intervals.
Following are examples of preparing compounds of formula I. The
stereochemistry listed is relative stereochemistry unless otherwise
noted. The terms cis and trans refer to the relative orientations
at the azetidinone 3- and 4-positions unless otherwise indicated.
The term "J" refers to the proton NMR coupling constant in hertz
(Hz) between the 3- and 4-substituted protons of the azetidinone.
All NMR data is of CDCl.sub.3 solution unless otherwise indicated.
##STR28##
Freshly prepare a solution of lithium diisopropylamide (LDA) by
dissolving diisopropylamine (1.19 g, 11.8 mmol) in anhydrous THF
(20 ml) at -78.degree. C. under argon. Add n-butyllithium (4.9 ml,
11.8 mmol, 2.4M in hexanes) and stir for 0.5 h at -78.degree. C. To
this cold solution add, 4phenylbutyrolactone (1.75 g, 10.8 mmol) in
THF (4 ml) over 0.25 h, keeping the reaction temperature below
-65.degree. C. Stir at -78.degree. C. for 0.25 h, then add
4-methoxybenzylidine anisidine (2.33 g, 11.0 mmol) in THF (8 ml)
over 1 h at -78.degree. C. Warm the reaction slowly to -50.degree.
C. over 1 h. Quench the reaction at low temperature with 1N HCl (12
ml). Partition the reaction mixture between ether and 1N HCl, wash
the ether layer with water, combine the ether extracts, dry over
MgSO.sub.4 and concentrate in vacuo. Crystallize the crude reaction
residue (3.0 g) from EtOAc-ether to obtain 1.54 g of compound A.
Reconcentrate the filtrate and chromatograph on silica gel 60,
eluting with 4:1 EtOAc-hexane, and isolate additional compound A
(0.385 g) as well as compound B (0.420 g).
Compound A: mp 218.degree.-220.degree. C.; IR 1730 cm-1; CI (M-H)
374; J=5.9 Hz.
Compound B: mp 74.degree.-76.degree. C.; IR 1730 cm-1; CI (M+H)
374; J=2.3 Hz.
Using a similar procedure and appropriate starting materials,
prepare compound 1C:
##STR29## Cl(M + H) 464, J = 2.3 Hz
EXAMPLE 2
##STR30##
To a solution of compound A from Example 1 (0.5 g, 1.3 mmol) in
anhydrous pyridine (2.7 ml), add acetic anhydride (0.63 ml, 6.7
mmol). Stir for 16 h, dilute with CH.sub.2 Cl.sub.2 and wash
3.times. with 1N HCl 1.times. with NaCI (sat'd) and 1.times. with
water. Concentrate the organic layer to dryness and crystallize the
residue from EtOAc to obtain the title compound (0.46 g), mp
167.degree.-169.degree. C.; IR 1745 cm-1; EI (M+) 415; J=5.9
Hz.
EXAMPLE 3
##STR31##
Freshly prepare a solution of lithium isopropylcyclohexylamide
(LICA) by adding n-butyllithium (2.84 mL of a 1.6M solution) to 5 a
solution of isopropylcyclohexylamine (0.75 mL) in THF (100 mL) at
-78.degree. C. Dissolve N-phenyl-4-(4-methoxyphenyl)-2-azetidinone
(1.0 g) in THF (8 mL) and slowly add to the LICA solution at
-78.degree. C. After stirring for 20 min, add hydrocinnamaldehyde
(0.54 g) and stir the reaction mixture at -78.degree. C. for 4 h.
Quench the reaction with 10% KHSO.sub.4 and extract the product
with EtOAc. Separate the organic layer, wash with water and NaCl
(sat'd). Concentrate the extract and purify the resultant residue
on a silica gel 60 column, eluting with EtOAc:hexane (15:85) to
obtain 1.15 g of product as a mixture of diastereomers. Separate
the diastereomers by HPLC on a silica gel column to give three
diastereomers 3A, 3B and 3C:
3A ##STR32## 1H in CDCl.sub.3 :7.32-7.18(m, 11H); 7.08-6.99 (m,
1H); 6.89(d, J = 9 Hz, 2H); 4.80(d, J = 2.4 Hz, 1H); 4.10-4.00(m,
1H); 3.79(s, 3H); 3.20-3.16(m, 1H); 2.90-2.67(m, 2H); 2.15-1.85(m,
3H) 3B ##STR33## 1H in CDCl.sub.3 :7.35-7.10(m, 11H); 7.08-6.99 (m,
1H); 6.89(d, J = 9 Hz, 2H); 5.09(d, J = 2.4 Hz, 1H); 4.26-4.14(m,
1H); 3.79(s, 3H); 3.21-3.14(m, 1H); 2.89-2.57(m, 2H); 2.10-1.85(m,
3H) 3C ##STR34## 1H in CDCl.sub.3 :7.30-7.00(m, 10H); 6.99 (d, J =
8 Hz, 2H); 6.83(d, J = 9 Hz, 2H); 5.12(d, J = 5.5 Hz, 1H); 3.82(s,
3H); 3.75-3.63(m, 1H); 3.52(dd, J = 9.5 Hz, 1H); 2.71-2.57(m, 1H);
2.49-2.33(m, 1H); 1.68-1.50(m, 1H); 1.47-1.31)m, 1H)
The 3A, 3B and 3C, diastereomers were further separated according
to the following reaction scheme, wherein partial structures are
shown: ##STR35##
(The following CD spectra data [.theta.] are all obtained in
CH.sub.3 OH.)
3D) [.theta.].sub.227nM =+2.0.times.10.sup.4 cm.sup.2 /dM;
[.theta.].sub.241nM =-4.6.times.10.sup.4 cm.sup.2 /dM. Elemental
analysis calc for C.sub.25 H.sub.25 NO.sub.3 -0.25 H.sub.2 O: C
76.6; H 6.56 N 3.57. found: C 76.66; H 6.49; N 3.64.
3E) [.theta.].sub.227nM =-1.95.times.10.sup.4 cm.sup.2 /dM;
[.theta.].sub.241nM =+4.45.times.10.sup.4 cm.sup.2 /dM. Elemental
analysis calc for C.sub.25 H.sub.25 NO.sub.3.0.5 H.sub.2 O: C
75.73; H 6.61; N 3.53. found: C 75.66; H 6.41; N 3.60.
3F) [.theta.].sub.226nM =+1.97.times.10.sup.4 cm.sup.2 /dM;
[.theta.].sub.240nM =-5.22.times.10.sup.4 cm.sup.2 /dM. Elemental
analysis calc for C.sub.25 H.sub.25 NO.sub.3 : C 77.48; H 6.5-1; N
3.62. found: C 77.44; H 6.53; N 3.70.
3G) [.theta.].sub.226nM =-1.78.times.10.sup.4 cm.sup.2 /dM;
[.theta.].sub.241nM =+4.78.times.10.sup.4 cm.sup.2 /dM (CIMS 388
M.sup.+ H).
3H) [.theta.].sub.226nM =+2.24.times.10.sup.4 cm.sup.2 /dM;
[.theta.].sub.241nM =-5.4.times.10.sup.4 cm.sup.2 /dM.
[.alpha.].sub.D.sup.25 =-54.4.degree. (2.5 mg/ml CH.sub.3 OH)
Elemental analysis calc for C.sub.25 H.sub.25 NO.sub.3 : C 77.48; H
6.51; N 3.62. found: C 77.11; H 6.50; N 3.72.
3I) [.theta.].sub.226nM =-2.05.times.10.sup.4 cm.sup.2 /dM;
[.theta.].sub.241nM =+5.2.times.10.sup.4 cm.sup.2 /dM. (CIMS 388
M.sup.+ H). ##STR36##
Add DEAD (0.11 ml) to a solution of compound 3H (132 mg), PPh.sub.3
(0.18 g) and HCO.sub.2 H (39 ml) in THF (5 ml). Stir at room
.[.temperatq.]. .Iadd.temperature .Iaddend.overnight, then
partition the reaction mixture between Et.sub.2 O and H.sub.2 O.
Wash (brine) and dry (MgSO.sub.4) the organic layer and concentrate
to dryness. Flash chromatograph the residue using EtOAc:Hex (1:4)
to obtain the formate ester. Dissolve this in CH.sub.3 OH and add 4
drops of conc. HCl. After 4 h, concentrate in vacuo and flash
chromatograph the residue using EtOAc:Hex (1:3) to obtain 3J.
[.theta.].sub.224nM =+2.54.times.10.sup.3 cm.sup.2 /dM;
[.theta.].sub.239nM =+5.70.times.10.sup.4 cm.sup.2 /dM;
[.alpha.].sub.D.sup.20 =-157.6.degree. (2.5 mg/ml CH.sub.3 OH)
##STR37##
Using the procedure described for 3J, treat compound 3I to obtain
3K. [.theta.].sub.222nM =-3.4.times.10.sup.3 cm.sup.2 /dM;
[.alpha.].sub.240nM =-5.6.times.10.sup.4 cm.sup.2 /dM.
[.alpha.].sub.D.sup.20 =+167.2.degree. (2.5 mg/ml CH.sub.3 OH)
Using the procedure described above for preparing compounds 3A and
3B, treat N-phenyl-4-(4-methoxyphenyl)-2-azetidinone with LICA
followed by 2-naphthaldehyde to obtain the diastereomers 3L and
3M:
3L ##STR38## mp 137- 138.degree. C. 3M ##STR39## mp 150-
151.degree. C.
EXAMPLE 4
##STR40##
Method 1:
Step 1) To a refluxing solution of of 4-methoxyberizylidene
anisidine (10.0 g, 41.5 mmol) and tributylamine (20.8 ml, 87 mmol)
in toluene (100 ml), add 5-bromovaleroyl chloride (8.5 g, 43, mmol)
in toluene (20 ml) dropwise over 2 h. Stir the reaction mixture at
80.degree. C. for 12 h, cool to room temperature, wash 3.times.
with 1 N HCl, 1.times. with water and dry the organic layer over
MgSO.sub.4. Purify by silica gel chromatography, eluting with ethyl
acetate:hexane (4:1) to obtain 5.1 g of (3R,
4S)-1,4-bis(4-methoxyphenyl)-3-(3-bromoproyl)-2-azetidinone
(relative stereochemistry), mp 70.degree.-73.degree. C., El
(M.sup.+) 404; J=2.3 Hz.
Step 2) To a solution of the product of step 1 (5.1 g, 12.6 mmol)
in (CH.sub.3).sub.2 SO (20 ml), add (CH.sub.3).sub.3 N(O) (2.39 g,
31.9 mmol). Heat the mixture at 60.degree. C. for 3 h, cool to room
temperature, dilute with EtOAc, and wash 3.times. with water.
Combine the aqueous fractions and extract with EtOAc. Combine the
organic fractions and concentrate. Purify the crude product by
silica gel chromatography, eluting with EtOAc:hexane (1:1) to
obtain 1.4 g (3R,
4S)-1,4-bis-(4-methoxyphenyl)-2-oxo-3-azetidine-propanol (relative
stereochemistry), an oil; EI (M.sup.+) 339; J=2.3 Hz.
Step 3) To a solution of the product of step 2 (.[.0.7134.].
.Iadd.0.734 .Iaddend.g, 2.2 mmol) in THF (4 ml) at 0.degree. C.,
add phenylmagnesium bromide (2.4 ml, 2.4 mmol, 1.0 M in THF) over
0.25 h. After 1 h at 0.degree. C., add water (5 ml), separate the
layers, wash the organic layer 1 .times. with 1N HCl, dry with
MgSO.sub.4 and concentrate to an oil. Purify by silica gel
chromatography, eluting with EtOAc:hexane (2:1) to obtain 0.372 g
of the title compound (mix of diastereomers) as an oil. CI (M.sup.+
H) 418.
Separation of diastereomers: Apply the diastereomeric mixture from
step 3 to a Chiralcel OD (Chiral Technologies Corp, Pa.)
chromatography column, eluting with hexane:ethanol (9:1) to obtain
enantiomerically pure (>98%) diastereomers as follows:
##STR41##
Method 2:
Step 1) To a solution of
1,4-(S)-bis(4-methoxyphenyl)-3-(3(R)-phenylpropyl)-2-azetidinone
(5.04 g, 0.013 mole) in CCl.sub.4 (20 ml) at 80.degree. C., add NBS
(2.76 g, 0.0155 mole) and benzoyl peroxide (0.24 g, 1.0 mmole) in
three equal portions over 1 h. Follow the reaction by TLC (4:1
hexane:EtOAc). Cool the reaction to 22.degree. C., add NaHSO.sub.4,
separate the layers and wash the organic layer 3.times. with water.
Concentrate the organic layer to obtain the crude product.
CI (M.sup.+ H) 480; .sup.1 H in CDCl.sub.3 .delta. PhCH(OH)=5.05
ppm.
Step 2) Dissolve the crude product of Step 1 in CH.sub.2 Cl.sub.2
(30 ml) and add 40% n-BuNOC(O)CF.sub.3 in water (30 ml). Reflux the
biphasic reaction for 24 h, cool, separate the layers and wash the
organic layer 6.times. with water. Concentrate the organic layer to
dryness and immediately redissolve the residue in ethanol saturated
with NH.sub.3 (10 ml). After 1 h, concentrate the reaction mixture
and partially purify by silica gel chromatography. Further purify
by HPLC to obtain a 1:1 mixture of compounds 4A and 4B. The mixture
can be further purified on a Chiracel OD column to obtain 4A and 4B
separately as characterized above.
Using the procedure described in Example 4, Method 2, with
4(S)-(4-acetoxyphenyl)-3(R)-(3-phenylpropyl)-1-(4-methoxy-phenyl)-2-azetid
inone as the starting material, prepare the following compounds:
##STR42##
EXAMPLE 5
##STR43##
To a solution of the product of step 2 of Example 4 (0.230 g, 0.68
mmol) in THF (2 ml), add the reagent derived from treatment of
4-methoxymethylphenyl bromide (0.159 g, 0.736 mmol) in THF (4 ml)
at -78.degree. C. with sec-butyllithium (0.6 ml, 0.78 mol, 1.3M in
hexanes), followed by CeCl.sub.3 (0.186 g, 0.75 mmol). After 4 h,
extract the product and purify by chromatography in a manner
similar to that described in step 3 of Example 4 to obtain 0.05 g
of the title compound (mix of diastereomers) as an oil. CI (M.sup.+
H) 478.
EXAMPLE 6
##STR44##
Step 1): To a solution of (S)-4-phenyl-2-oxazolidinone (41 g, 0.25
mol) in CH.sub.2 Cl.sub.2 (20 ml), add 4-dimethylaminopyridine (2.5
g, 0.02 mol) and triethylamine (84.7 ml, 0.61 mol) and cool the
reaction to 0.degree. C. Add methyl-4-(chloroformyl)butyrate (50
.[.9.]. .Iadd.g.Iaddend., 0.3 mol) as a solution in CH.sub.2
Cl.sub.2 (375 ml) dropwise over 1 h, and allow the reaction to warm
to 22.degree. C. After 17 h, add water and H.sub.2 SO.sub.4 (2N,
100 ml), separate the layers, and wash the organic layer
sequentially with NaOH (10%). NaCI (sat'd) and water. Dry the
organic layer over MgSO.sub.4 and concentrate to obtain a
semicrystalline product.
Step 2): To a solution of TiCl.sub.4 (18.2 ml, 0.165 mol) in
CH.sub.2 Cl.sub.2 (600 ml) at 0.degree. C., add titanium
isopropoxide (16.5 ml, 0.055 mol). After 15 min, add the product of
Step 1 (49.0 9, 0.17 mol) as a solution in CH.sub.2 Cl.sub.2 (100
ml). After 5 min., add diisopropylethylamine (DIPEA) (65.2 ml, 0.37
mol) and stir at 0.degree. C. for 1 h, cool the reaction mixture to
-20.degree. C., and add 4-benzyloxybenzylidine(4-fluoro)aniline
(114.3 g, 0.37 mol) as a solid. Stir the reaction vigorously for 4
h at -20.degree. C., add acetic acid as a solution in CH.sub.2
Cl.sub.2 dropwise over 15 min, allow the reaction to .[.warnr.].
.Iadd.warm .Iaddend.to 0.degree. C., and add H.sub.2 SO.sub.4 (2N).
Stir the reaction an additional 1 h, separate the layers, wash with
water, separate and dry the organic layer. Crystallize the crude
product from ethanol/water to obtain the pure intermediate.
Step 3): To a solution of the product of Step 2 (8.9 g, 14.9 mmol)
in toluene (100 ml) at 50.degree. C., add
N,O-bis(trimethylsilyl)acetamide (BSA) (7.50 ml, 30.3 mmol). After
0.5 h, add solid TBAF (0.39 g, 1.5 mmol) and stir the reaction at
50.degree. C. for an additional 3 h. Cool the reaction mixture to
22.degree. C., add CH.sub.3 OH (10 ml), wash the reaction mixture
with HCl (1N), NaHCO.sub.3 (1N) and NaCI (sat'd), and dry the
organic layer over MgSO.sub.4.
Step 4): To a solution of the product of Step 3 (0.94 g, 2.2 mmol)
and CH.sub.3 OH (3 ml), add water (1 ml) and LiOH.H.sub.2 O (102
mg, 2.4 mmole). Stir the reaction at 22.degree. C. for 1 h and add
additional LiOH.H.sub.2 O (54 mg, 1.3 mmole). After a total of 2 h,
add HCl (1N) and EtOAc, separate the layers, dry the organic layer
and concentrate in vacuo. To a solution of resultant product (0.91
g, 2.2 mmol) in CH.sub.2 Cl.sub.2 at 22.degree. C., add ClCOCOCl
(0.29 ml, 3.3 mmol) and stir for 16 h. Remove the solvent in
vacuo.
Step 5): To an efficiently stirred suspension of 4-fluorophenylzinc
chloride (4.4 mmol) prepared from 4-fluorophenylmagnesium bromide 5
(1M in THF, 4.4 ml, 4.4 mmol) and ZnCl.sub.2 (0.6 g, 4.4 mmol) at
4.degree. C., add tetrakis(triphenylphosphine)palladium (0.25 g,
0.21 mmol) and the product of Step 4 (0.94 g, 2.2 mmol) as a
solution in THF (2 ml). Stir the reaction for 1 h at 0.degree. C.
and then for 0.5 h at 22.degree. C. Add HCl (1N, 5 ml) and extract
with EtOAc. Concentrate the organic layer to an oil and purify by
silica gel chromatography to obtain
1-(4-fluorophenyl)-4(S)-(4-hydroxyphenyl)-3(R)-(3-oxo-3-phenylpropyl)-2-az
etidinone:
HRMS calc'd for C.sub.24 H.sub.19 F.sub.2 NO.sub.3 =408.1429, found
408.1411.
Step 6): To the product of Step 5 (0.95 g, 1.91 mmol) in THF (3
ml), add
(R)-tetrahydro-1-methyl-3,3-diphenyl-1H,3H-pyrrolo-[1,2-c][1,3,2]oxazaboro
le (120 mg, 0.43 mmol) and cool the mixture to -20.degree. C. After
5 min, add borohydride-dimethylsulfide complex (2M in THF: 0.85 ml,
1.7 mmol) dropwise over 0.5 h. After a total of 1.5 h, add CH.sub.3
OH followed by HCl (1 N) and extract the reaction mixture with
EtOAc to obtain
1-(4-fluorophenyl)-3(R)-[3(S)-(4-fluorophenyl)-3-hydroxypropyl)]-4(S)-[4-(
phenylmethoxy)phenyl]-2-azetidinone (compound 6A-1) as an oil.
.sup.1 H in CDCl.sub.3 .delta. H3=4.68, J=2.3 Hz. CI (M.sup.+ H)
500.
Use of
(S)-tetra-hydro-1-methyl-3,3-diphenyl-1H,3H-pyrrolo-[1,2-c][1,3,2]
oxazaborole gives the corresponding 3(R)-hydroxypropyl azetidinone
(compound 6B-1). .sup.1 H in CDCl.sub.3 .delta. H3=4.69, J=2.3 Hz.
CI (M.sup.+ H) 500.
To a solution of compound 6A-1 (0.4 g, 0.8 mmol) in ethanol (2 ml),
add 10% Pd/C (0.03 g) and stir the reaction under a pressure (60
psi) of H.sub.2 gas for 16 h. Filter the reaction mixture and
concentrate the solvent to obtain compound 6A. Mp
164.degree.-166.degree. C.; CI (M.sup.+ H) 410.
[.alpha.].sub.D.sup.25 =-28.1.degree. (c 3, CH.sub.3 OH). Elemental
analysis calc'd for C.sub.24 H.sub.21 F.sub.2 NO.sub.3 ; C 70.41; H
5.17; N 3.42; found C 70.25; H 5.19; N 3.54.
Similarly treat compound 6B-1 to obtain compound 6B. Mp
129.5.degree.-132.5.degree. C.; CI (M.sup.+ H) 410. Elemental
analysis calc'd for C.sub.24 H.sub.21 F.sub.2 NO.sub.3 : C 70.41; H
5.17; N 3.42; found C 70.30; H 5.14; N 3.52.
Step 6') (Alternative): To a solution of the product of Step 5
(0.14 g, 0.3 mmol) in ethanol (2 ml), add 10% Pd/C (0.03 g) and
stir the reaction under a pressure (60 psi) of H.sub.2 gas for 16
h. Filter the reaction mixture and concentrate the solvent to
afford a 1:1 mixture of compounds 6A and 6B.
Using appropriate starting materials and following the procedure of
steps 1-6, prepare the following compounds: ##STR45##
EXAMPLE 7
##STR46##
To a solution of 7a (1.0 g, 2.1 mmol) in dioxane (10 ml), add
SeO.sub.2 (1.33 g, 11.98 mmol) and water .[.(25 ml, 14 mmol),.].
.Iadd.(0.25 ml, 14 mmol), .Iaddend.and heat the reaction to
100.degree. C. After 1 h, cool the reaction to room temperature and
isolate by extraction the crude product as a diastereomeric mixture
(1:2) of alcohols 7b-A and 7b-B. Purify by HPLC on a Dynamax silica
column to separate diastereomers 7b-A and 7b-B.
Diastereomer 7b-A (R): oil; J.sub.34 =2.3 Hz, .delta. C H(OH)=4.86
(t); HRMS C.sub.32 H.sub.29 NO.sub.4 calc.: 491.2097; found:
491.2074.
Diastereomer 7b-E (S): oil; J.sub.34 =2.3 Hz, .delta. C H(OH)=5.06
(t.Iadd.).Iaddend.; HRMS C.sub.32 H.sub.29 NO.sub.4 calc.:
491.2097; found: 491.2117.
Step 2): To a solution of diastereomer A from step 1 (58 mg, 0.12
mmol) in EtOAc (2 ml), add 10% Pd on carbon (20 mg) and stir at
22.degree. C. under H.sub.2 gas (14 psi) for 12 h. Filter and
concentrate to obtain the title compound as a semisolid, m.p.
90.degree.-92.degree. C. J.sub.34 =2.3 Hz, .delta. CH(OH)=4.1 (m);
HRMS C.sub.25 H.sub.25 NO.sub.4 calc: 403.1783; found:
403.1792.
EXAMPLE 8
To a solution of the product of Example 4A (90 mg, 0.2 mmol) in
CH.sub.2 Cl.sub.2, add acetyl chloride (80 mg, 1.0 mmol) and
pyridine (8 mg, 0.1 mmol) and stir at room temperature for 1 h. Add
water, separate the layers and isolate the corresponding acetoxy
compound, 8A. In a similar manner, treat the products of Examples
4B, 6B and 6A to obtain the following compounds 8B, 8.degree. C.
and 8D, respectively:
8A:
1,4(S)-bis(4-methoxyphenyl)-3(R)-(3(R)-acetoxy-3-phenylpropyl)-2-azetidino
ne. CI (M.sup.+ H) 460; HRMS C.sub.28 H.sub.29 NO.sub.5 calc.:
459.2044; found: 459.2045.
8B:
1,4(S)-bis(4-methoxyphenyl)-3(R)-(3(S)-acetoxy-3-phenylpropyl)-2-azetidino
ne. CI (M.sup.+ H) 460; HRMS C.sub.28 H.sub.29 NO.sub.5 calc.:
459.2044; found 459.2048.
8C: 4(S)-(4-acetyloxyphenyl)-3(R)-(3(R)-acetylox.[.i.].
.Iadd.y.Iaddend.
-3-(4-fluorophenyl)propyl)-1-(4-fluorophenyl)-2-azetidinone. FAB
M:S 493.4; HRMS C.sub.28 H.sub.25 F.sub.2 NO.sub.5 calc.: 493.1695;
found: 493.1701.
8D:
4(S)-(4-acetyloxyphenyl)-3(R)-(3(S)-acetyloxy-3-(4-fluorophenyl)propyl)-1-
(4-fluorophenyl)-2-azetidinone. FAB MS 493.4; HRMS C.sub.28
H.sub.25 F.sub.2 NO.sub.5 calc.: 493.1695; found: 493.1694.
Using appropriate starting materials in the procedure of Example 6,
prepare 1-(4-chlorophenyl)-3(R)-(hydroxy-3.[...]. .Iadd.-.Iaddend.
(4-chlorophenylpropyl)-4(S)-(4-hydroxyphenyl)-2-azetidinone. Using
the procedure of Example 8, prepare the following diacetates 8E and
8F: ##STR47##
EXAMPLE 9
##STR48##
Step 1:
Add pyridinium chlorochromate (2.4 g, 11 mmoles) and CH.sub.3
CO.sub.2 Na (approx. 20 mg) to a solution of
1-phenyl-3-(3-phenyl-1-hydroxypropyl)-4-(4-methoxyphenyl)-2-azetidinone
(2.35 g, 6.1 mmoles) in CH.sub.2 Cl.sub.2. Stir at room temperature
for 18 h, then add silica gel (40 g) and concentrate to dryness.
Flash chromatograph the residue using EtOAc:Hex (1:4) to obtain an
oil. (1.98 g, yield=85%). .sup.1 H NMR 2.85-2.95 (m, 3H), 3.15 (m,
1H), 3.80 (s, 3H), 4.10 (d, 1H, J 2.6), 5.42 (1H, d, 6.85 (dd, 2H,
J 2.8), 7.05 (m, 1H), 7.2-7.35 (m, 11H).
Step 2:
To a solution of the product of Step 1 (1.78 g, 4.62 mmoles) in THF
at -10.degree. C., add NaH (115 mg, 4.8 mmoles). After 15 min, add
NBS (865 mg, 4.85 mmoles) and stir for 20 min., then add 1N HCl and
partition between EtOAc and brine. Separate the organic layer, dry
(MgSO.sub.4) and concentrate to give an oil. Flash chromatograph
the oil using EtOAc:Hex (1:10) to collect first 9a as a foamy solid
(830 mg, y=39%, FAB MS 466/464, M+H), and then 9b as a colorless
solid (1.1 g, y=51%, FAB MS 466/464, M+H).
Step 3a:
Add Mg(OCOCF.sub.3).sub.2.CF.sub.3 CO.sub.2 H (7.3 ml of 1M
solution is Et.sub.2 O.) to a solution of 9a (0.68 g, 1.46 mmoles)
in THF (5 ml) at -50.degree. C. Stir the reaction 5 min., then add
t-Bu--NH.sub.2 -BH.sub.3 (254 mg, 2.92 mmole). After 15 min., allow
the reaction to warm to 0.degree. C. over 20 min., add 1N HCl and
concentrate in vacuo. Partition the residue between EtOAc and
brine. Concentrate the organic layers and dissolve the resultant
oil in CH.sub.2 Cl.sub.2 :CH.sub.3 OH (1:1) and add ethanolamine
(approx 2 mmoles). After 15 min., concentrate the reaction mixture
and partition the residue with EtOAc:1N HCl. Wash (brine) and dry
(MgSO.sub.4) the organic layer to obtain an oil. Purify this oil by
flash chromatography using EtOAc:Hex (1:4) to obtain compound 9a-1,
a colorless solid, as a 4:1 mix of diastereomers. 0.52 g, y=76%,
SIMS 468/466 (M+H).
Step 3b:
Using compound 9b as the starting material, use a procedure similar
to Step 3a with CH.sub.2 Cl.sub.2 as solvent for the preparation of
9b-1 in 80% yield as a 13:1 mixture of diastereomers (SIMS 468/466
M+H).
Step 4a:
Add a solution of 9a-1 (0.27 g, 0.58 mmoles) and AIBN (18 mg, 0.12
mmole) in toluene (40 ml) dropwise over 40 min. to a solution of
(TMS).sub.3 SiH (1.0 ml) in toluene at 80.degree. C. for 1.5 h.
Cool and concentrate the reaction mixture, dissolve the residue in
CH.sub.3 CN and wash 3.times. with hexane. Concentrate the CH.sub.3
CN layer to give the title compound as a racemic mixture (0.25 g).
Purify this oil by HPLC using a Chiralcel OD column to obtain 3H
(major) and 3J (minor).
Step 4b:
Use the procedure of Step 4a, starting with compound 9b-1 to obtain
an oil. Purify this by flash chromatography using EtOAc:Hex (1:3)
to collect the racemic title compound (y=70%). Purify this oil by
HPLC using a Chiralcel OD column to obtain 3J (major) and 3H
(minor).
EXAMPLE 10
##STR49##
Step 1:
Follow the procedure of Example 3, using
1-(4-fluorophenyl-4-(4-t-butyldimethylsilyloxyphenyl)-2-azetidinone
to obtain 1-(4-fluorophenyl-3-(3-phenyl-1-hydroxypropyl)
4-(4-t-butyldimethylsilyl-oxyphenyl)-2-azetidinone.
Step 2:
Treat a solution of the cis-azetidinone of Step 1 (0.25 g) in
.[.CH3CN.]. .Iadd.CH.sub.3 CN .Iaddend.(21 ml) with 48% aqueous HF
(2.5 ml). After 18 h, dilute the reaction mixture with cold H.sub.2
O and extract with Et.sub.2 O. Wash (2.times. H.sub.2 O, dilute
NaHCO.sub.3 and brine), dry (MgSO.sub.4) and concentrate the
Et.sub.2 O layer. Crystallize the residue from EtOAc:hexane (1:2)
to obtain the title compound as colorless needles (123 mg, y=64%),
mp 168.degree.-171.degree. C. Elemental analysis calc for C.sub.24
H.sub.22 O.sub.3 FN: C 73.64; H 5.66; N 3.58. found C 73.32; H
5.65; N 3.68.
The following formulations exemplify some of the dosage of this
invention. In each the term "active compound" designates a compound
of formula I.
EXAMPLE A
Tablets No. Ingredient mg/tablet mg/tablet 1 Active Compound 100
500 2 Lactose USP 122 113 3 Corn Starch, Food Grade, as a 10% 30 40
paste in Purified Water 4 Corn Starch, Food Grade 45 40 5 Magnesium
Stearate 3 7 Total 300 700
Method of Manufacture
Mix Item Nos. 1 and 2 in suitable mixer for 10-15 minutes.
Granulate the mixture with Item No. 3. Mill the damp granules
through a coarse screen (e.g., 1/4', 0.63 cm) if necessary. Dry the
.[.cl.]. .Iadd.d.Iaddend.amp granules. Screen the dried granules if
necessary and mix with Item No. 4 and mix for 10-15 minutes. Add
Item No. 5 and mix for 1-3 minutes. Compress the mixture to
appropriate size and weight on a suitable tablet machine.
EXAMPLE B
Capsules No. Ingredient mg/tablet mg/tablet 1 Active Compound 100
500 2 Lactose USP 106 123 3 Corn Starch, Food Grade 40 70 4
Magnesium Stearate NF 4 7 Total 250 700
Method of Manufacture
Mix Item Nos. 1, 2 and 3 in a suitable blender for 10-15 minutes.
Add Item No. 4 and mix for 1-3 minutes. Fill the mixture suitable
two-piece hard gelatin capsules on a suitable encapsulating
machine.
Representative formulations comprising a cholesterol biosynthesis
inhibitor are well known in the art. It is contemplated that where
the two active ingredients are administered as a single
composition, the dosage forms disclosed above for substituted
azetidinone compounds may readily be modified using the knowledge
of one skilled in the art.
Using the test procedures described above, the following in vivo
data were obtained for the exemplified compounds. Data is reported
as percent change (i.e., percent reduction in cholesterol esters)
versus control, therefore, negative numbers indicate a positive
lipid-lowering effect.
% Reduction Serum Cholest. Dose Ex. # Cholest. Esters mg/kg 1A -23
0 50 1B -15 -39 50 1C 14 0 50 2 0 0 50 3A -31 -69 50 3C -60 -92 50
3D -17 -61 10 3E 0 0 10 3F -29 -77 10 3G -16 -38 10 3H -41 -86 10
3I 0 -22 10 3J 0 0 3 3K 0 0 10 3L -15 -21 10 3M 0 -22 10 4A 0 -54 5
4B -37 -89 8 4C -12.5 0 3 4D 9 0 7 4E 0 -46 3 4F -29 -95 3 5 0 -64
10 6A -59 -95 1 6A-1 -43 -93 1 6B -40 -92 3 6C 0 -48 3 6D -46 -95
10 8A 0 -44 3 8B -50 -95 3 8C -14 -37 1 8D -49 -98 1 8E -22 -66 3
8F -43 -94 1 10 -26 -77 3
* * * * *