U.S. patent number RE36,481 [Application Number 09/188,158] was granted by the patent office on 2000-01-04 for hmg-coa reductase inhibitors.
This patent grant is currently assigned to Merck & Co., Inc.. Invention is credited to Wasyl Halczenko, George D. Hartman, Otto D. Hensens, David R. Houck, Edward S. Inamine, Ta J. Lee, Robert L. Smith, Gerald E. Stokker.
United States Patent |
RE36,481 |
Inamine , et al. |
January 4, 2000 |
HMG-CoA reductase inhibitors
Abstract
Novel 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase
inhibitors are useful as antihypercholesterolemic agents and are
represented by the following general structural formulae (I) and
(II): ##STR1##
Inventors: |
Inamine; Edward S. (Rahway,
NJ), Hensens; Otto D. (Red Bank, NJ), Houck; David R.
(Los Alamos, NM), Lee; Ta J. (Lansdale, PA), Smith;
Robert L. (Lansdale, PA), Halczenko; Wasyl (Hatfield,
PA), Hartman; George D. (Lansdale, PA), Stokker; Gerald
E. (Gwynedd Valley, PA) |
Assignee: |
Merck & Co., Inc. (Rahway,
NJ)
|
Family
ID: |
27535033 |
Appl.
No.: |
09/188,158 |
Filed: |
November 4, 1998 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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092393 |
Sep 2, 1987 |
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048136 |
May 15, 1987 |
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001933 |
Oct 9, 1987 |
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877041 |
Jun 23, 1986 |
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Reissue of: |
254525 |
Oct 6, 1988 |
04940727 |
Jul 10, 1990 |
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Current U.S.
Class: |
514/450; 514/460;
549/292 |
Current CPC
Class: |
C07C
69/732 (20130101); C07D 309/30 (20130101) |
Current International
Class: |
C07C
69/732 (20060101); C07D 309/30 (20060101); C07D
309/00 (20060101); C07C 69/00 (20060101); A61K
031/335 (); C07D 315/00 () |
Field of
Search: |
;549/292
;514/450,460 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
Other References
G E. Stokker et al., "3-Hydroxy-3-methylglutaryl-coenzyme A
Reductase Inhibitors. 1. Structural Modification of 5-Substituted
3,5, Dihydroxypentanoic Acids and Their Lactone Derivatives", J.
Med. Chem., 28, pp. 347-358 (1985). .
R. A. Heller et al., "3-Hydroxy-3-methylglutaryl Coenzyme A
Reductase from Rat Liver", J. Biol. Chem., 251, pp. 3815-3822
(1976). .
D. A. Kleinsek et al., "Purification of
3-hydroxy-3-methylglutaryl-coenzyme A reducatase from rat liver",
Proc. Nat'l Acad. Sci. USA, 74, pp. 1431-1435 (1977). .
S. Shefer et al., "HMG CoA reductase of intestinal mucosa and liver
of the rat", J. Lipid Res., 13, pp. 402-412 (1972). .
Sugiyama et al., Chemical Abstracts, 111, Abstract No. 120925n, pp.
366-367 (1989). .
A. F. Rosenthal et al., "Compounds Related to a-Glycerophosphoric
Acid, Phosphorylcholine and Phosphorylethanolamine", J. Amer. Chem.
Soc., 80, pp. 5240-5241 (1959). .
A. G. Olsson et al., "Synvinolin In Hypercholesterolaemia", The
Lancet, pp. 390-391 Aug. 16, 1986. .
P. Weisweiler et al., "Colestipol Plus Fenofibrate Versus
Synvinolin In Familial Hypercholesterolaemia", The Lancet, pp.
1212-1213, Nov. 22, 1986. .
A. F. Stalenhoef et al., "Effect of Synvinolin, a New Cholesterol
Synthesis Inhibitor, in Familial Hypercholesterolemia",
Arteriosclerosis, vol. 6, No. 5, Sep./Oct. 1986. .
M.J.T.M. Mol et al., "Effects of Synvinolin (MK-733) On Plasma
Lipids In Familial Hypercholesterolaemia", The Lancet, pp. 936-939,
Oct. 25, 1986. .
S. M. Grundy et al., "Effects of Mevinolin on Plasma Lipoprotein
(LP) Metabolism in Moderate Hypercholesterolemia", ABSTRACTS
Circulation, vol. 70, Supp. II, p. 268, No. 1072 (1984). .
D. W. Bilheimer et al., "Mevinolin and colestipol stimulate
receptor-mediated clearance of low density lipoprotein from plasma
in familial hypercholesterolemia heterozygotes", Proc. Natl. Acad.
Sci. USA, vol. 80, pp. 4124-4128 (Jul. 1983). .
D. W. Bilheimer et al., "Mevinolin Stimulates Receptor-Mediated
Clearance of LDL in Familial Hypercholesterolemia (FH)
Heterozygotes", Clin. Res. 31, No. 2, p. 544A (1983). .
J. A. Tobert et al., Cholesterol-lowering effect of Mevinolin, an
Inhibitor of 3-Hydroxy-3-Methylglutaryl-Coenzyme A Reductase, in
Healthy Volunteers, J. Clin. Invest. (USA), 69(4), PP. 913-919
(Apr. 1982). .
J. A. Tobert et al., "Rapid and Substantial Lowering of Human Serum
Cholesterol by Mevinolin (MK-803), An Inhibitor of
Hydroxymethylglutaryl-Coenzyme A Reductase", Atherosclerosis, 41,
pp. 61-65 (1982)..
|
Primary Examiner: Owens; Amelia
Attorney, Agent or Firm: Quagliato; Carol S. Winokur;
Melvin
Parent Case Text
This Application is a continuation-in-part of U.S. patent
application Ser. No. 092,353, filed Sept. 2, 1987 and U.S. patent
application Ser. No. 048,136 filed May 15, 1987, which is a
continuation-in-part of Ser. No. 001,933 filed Oct. 9, 1987, which
is a continuation-in-part of Ser. No. 877,041 filed June 23, 1986,
all now abandoned.
Claims
What is claimed is:
1. A compound represented by the following structural formula (I):
##STR49## wherein: R is ##STR50## R.sup.1 and R.sup.5 are
independently: (1) C.sub.1-10 alkyl;
(2) substituted C.sub.1-10 alkyl in which one or more
substituent(s) is:
(a) halogen,
(b) hydroxy,
(c) C.sub.1-10 alkoxy,
(d) C.sub.1-5 alkoxycarbonyl,
(e) C.sub.1-5 acyloxy,
(f) C.sub.3-8 cycloalkyl,
(g) phenyl,
(h) substituted phenyl in which the substituents are X and Y,
(i) C.sub.1-10 alkylS(O).sub.n in which n is 0 to 2,
(j) C.sub.3-8 cycloalkylS(O).sub.n,
(k) phenylS(O).sub.n,
(l) substituted phenylS(O).sub.n in which the substituents are X
and Y, and
(m) oxo;
(3) C.sub.1-10 alkoxy;
(4) C.sub.2-10 alkenyl;
(5) C.sub.3-8 cycloalkyl;
(6) substituted C.sub.3-8 cycloalkyl in which one substituent
is:
(a) C.sub.1-10 alkyl
(b) substituted C.sub.1-10 alkyl in which the substituent is
selected from
(i) halogen,
(ii) hydroxy,
(iii) C.sub.1-10 alkoxy,
(iv) C.sub.1-5 alkoxycarbonyl,
(v) C.sub.1-5 acyloxy,
(vi) phenyl,
(vii) substituted phenyl in which the substituents are X and Y
(viii) C.sub.1-10 alkylS(O).sub.n,
(ix) C.sub.3-8 cycloalkylS(O).sub.n,
(x) phenylS(O).sub.n,
(xi) substituted phenylS(O).sub.n in which the substituents are X
and Y, and
(xii) oxo,
(c) C.sub.1-10 alkylS(O).sub.n,
(d) C.sub.3-8 cycloalkylS(O).sub.n,
(e) phenylS(O).sub.n,
(f) substituted phenylS(O).sub.n in which the substituents are X
and Y,
(g) halogen,
(h) hydroxy,
(i) C.sub.1-10 alkoxy,
(j) C.sub.1-5 alkoxycarbonyl,
(k) C.sub.1-5 acyloxy,
(l) phenyl, and
(m) substituted phenyl in which the substituents are X and Y;
(7) phenyl;
(8) substituted phenyl in which the substituents are X and Y;
(9) amino;
(10) C.sub.1-5 alkylamino;
(11) di(C.sub.1-5 alkyl)amino;
(12) phenylamino;
(13) substituted phenylamino in which the substituents are X and
Y;
(14) phenyl C.sub.1-10 alkylamino;
(15) substituted phenyl C.sub.1-10 alkylamino in which the
substituents are X and Y; and R.sup.9 S in which R.sup.9 is
selected from
(a) C.sub.1-10 alkyl,
(b) phenyl, and
(c) substituted phenyl in which the substituents are X and Y;
R.sup.6 are independently:
(1) hydrogen;
(2) C.sub.1-5 alkyl;
(3) substituted C.sub.1-5 alkyl in which the substituent is:
(a) phenyl,
(b) dimethylamino, and
(c) acetylamino, and
(4) 2,3-dihydroxypropyl;
R.sup.3 and R.sup.4 are independently:
(1) hydrogen;
(2) C.sub.1-10 alkyl;
(3) substituted C.sub.1-10 alkyl in which one or more
substituent(s) is:
(a) halogen,
(b) hydroxy,
(c) C.sub.1-10 alkoxy,
(d) C.sub.1-5 alkoxycarbonyl,
(e) C.sub.1-5 acyloxy,
(f) C.sub.3-8 cycloalkyl,
(g) phenyl,
(h) substituted phenyl in which the substituents are X and Y,
(i) C.sub.1-10 alkylS(O).sub.n,
(j) C.sub.3-8 cycloalkylS(O).sub.n,
(k) phenylS(O).sub.n,
(l) substituted phenylS(O).sub.n in which the substituents are X
and Y, and
(m) oxo;
(4) C.sub.2-10 alkenyl;
(5) substituted C.sub.2-10 alkenyl in which one or more
substituent(s) is:
(a) halogen,
(b) hydroxy,
(c) C.sub.1-10 alkoxy,
(d) C.sub.1-5 alkoxycarbonyl,
(e) C.sub.1-5 acyloxy,
(f) C.sub.3-8 cycloalkyl,
(g) phenyl,
(h) substituted phenyl in which the substituents are X and Y,
(i) C.sub.1-10 alkylS(O).sub.n,
(j) C.sub.3-8 cycloalkylS(O).sub.n,
(k) phenylS(O).sub.n,
(l) substituted phenylS(O).sub.n in which the substituents are X
and Y, and
(m) oxo;
(6) C.sub.3-8 cycloalkyl;
(7) substituted C.sub.3-8 cycloalkyl in which one substituent
is:
(a) C.sub.1-10 alkyl
(b) substituted C.sub.1-10 alkyl in which the substituent is
(i) halogen,
(ii) hydroxy,
(iii) C.sub.1-10 alkoxy,
(iv) C.sub.1-5 alkoxycarbonyl,
(v) C.sub.1-5 acyloxy
(vi) phenyl,
(vii) substituted phenyl in which the substituents are X and Y
(viii) C.sub.1-10 alkylS(O).sub.n,
(ix) C.sub.3-8 cycloalkylS(O).sub.n,
(x) phenylS(O).sub.n in which the substituents are X and Y, and
(xii) oxo,
(c) C.sub.1-10 alkylS(O).sub.n,
(d) C.sub.3-8 cycloalkylS(O).sub.n,
(e) phenylS(O).sub.n,
(f) substituted phenylS(O).sub.n in which the substituents are X
and Y,
(g) halogen,
(h) hydroxy,
(i) C.sub.1-10 alkoxy,
(j) C.sub.1-5 alkoxycarbonyl,
(k) C.sub.1-5 acyloxy,
(l) phenyl, and
(m) substituted phenyl in which the substituents are X and Y;
(8) phenyl;
(9) substituted phenyl in which the substituents are X and Y;
R.sup.7 and R.sup.8 are independently:
(1) hydrogen;
(2) C.sub.1-10 alkyl;
(3) substituted C.sub.1-10 alkyl in which one or more
substituent(s) is:
(a) halogen,
(b) hydroxy,
(c) C.sub.1-10 alkoxy,
(d) C.sub.1-10 alkoxycarbonyl,
(e) C.sub.1-5 acyloxy,
(f) C.sub.3-8 cycloalkyl,
(g) phenyl,
(h) substituted phenyl in which the substituents are X and Y,
(i) C.sub.1-10 alkyl S(O).sub.n in which n is 0 to 2,
(j) C.sub.3-8 cycloalkyl S(O).sub.n,
(k) phenyl S(O).sub.n ;
(l) substituted phenyl S(O).sub.n in which the substituents are X
and Y, and
(m) oxo;
(4) C.sub.2-10 alkenyl;
(5) C.sub.3-8 cycloalkyl;
(6) aminocarbonyl;
(7) substituted aminocarbonyl in which one or more substituent(s)
is:
(a) C.sub.1-5 alkyl,
(b) C.sub.3-8 cycloalkyl,
(c) phenyl,
(d) substituted phenyl in which the substituents are X and Y;
(8) phenyl;
(9) substituted phenyl in which the substituents are X and Y;
(10) C.sub.1-10 alkylcarbonyl;
(11) C.sub.3-8 cycloalkylcarbonyl;
(12) phenylcarbonyl;
(13) substituted phenylcarbonyl in which the substituents are X and
Y; and
X and Y independently are hydrogen, halogen, trifluoromethyl,
C.sub.1-3 alkyl, nitro, cyano or a group selected from:
(1) R.sup.10 O (CH.sub.2).sub.m in which m is 0 to 3 and R.sup.10
is hydrogen, C.sub.2-3 alkyl or hydroxy C.sub.2-3 alkyl;
(2) ##STR51## in which R.sup.11 is hydrogen, C.sub.1-3 alkyl,
hydroxy-C.sub.2-3 alkyl, phenyl, naphthyl, amino C.sub.1-3 alkyl,
C.sub.1-3 alkylamino-C.sub.1-3 alkyl, di(C.sub.1-3
alkyl)amino-C.sub.1-3 alkyl, hydroxy C.sub.2-3 alkylamino-C.sub.1-3
alkyl or di(hydroxy C.sub.2-3 alkyl) amino C.sub.1-3 alkyl;
(3) ##STR52## in which R.sup.12 is hydrogen, C.sub.1-3 alkyl,
hydroxy-C.sub.2-3 alkyl, C.sub.1-3 alkoxy-C.sub.1-3 alkyl, phenyl
or naphthyl;
(4) ##STR53## in which R.sup.13 and R.sup.14 independently are
hydrogen, C.sub.1-3 alkyl, and hydroxy-C.sub.2-3 alkyl;
(5) R.sup.15 S(O).sub.n (CH.sub.2)m in which R.sup.15 is hydrogen,
C.sub.1-3 alkyl, amino, C.sub.1-3 alkylamino or di(C.sub.1-3
alkyl)amino; and
a, b and c each represent single bonds or one of a, b and c
represents a double bond or both a and c represent double bonds; or
a pharmaceutically acceptable salt thereof.
2. A compound of claim 1 wherein:
R.sup.1 and R.sup.5 are independently:
(1) C.sub.1-10 alkyl;
(2) substituted C.sub.1-10 alkyl in which one or more
substituent(s) is:
(a) halogen,
(b) hydroxy,
(c) C.sub.1-10 alkoxy,
(d) C.sub.1-5 alkoxycarbonyl,
(e) C.sub.1-5 acyloxy,
(f) C.sub.3-8 cycloalkyl,
(g) phenyl,
(h) substituted phenyl in which the substituents are X and Y,
and
(i) oxo;
(3) C.sub.3-8 cycloalkyl;
(4) substituted C.sub.3-8 cycloalkyl in which one substituent
is:
(a) C.sub.1-10 alkyl,
(b) substituted C.sub.1-10 alkyl in which the substituent is
(i) halogen,
(ii) hydroxy,
(iii) C.sub.1-10 alkoxy
(iv) C.sub.1-5 acyloxy,
(v) C.sub.1-5 alkoxycarbonyl,
(vi) phenyl,
(vii) substituted phenyl in which the substituents are X and Y,
and
(viii) oxo,
(c) halogen,
(d) hydroxy,
(e) C.sub.1-10 alkoxy,
(f) C.sub.1-5 alkoxycarbonyl,
(g) C.sub.1-5 acyloxy,
(h) phenyl,
(i) substituted phenyl in which the substituents are X and Y;
(5) phenylamino;
(6) substituted phenylamino in which the substituents are X and
Y;
(7) phenyl C.sub.1-10 alkylamino; and
(8) substituted phenyl C.sub.1-10 alkylamino in which the
substituents are X and Y.
3. A compound of claim 2 wherein:
R.sup.1 and R.sup.5 are independently:
(1) C.sub.1-10 alkyl;
(2) C.sub.3-8 cycloalkyl;
(3) phenylamino; and
(4) substituted phenylamino in which the substituents are X and
Y.
4. A compound of claim 3 wherein:
R.sup.3 and R.sup.4 are independently:
(1) hydrogen;
(2) C.sub.1-10 alkyl;
(3) C.sub.3-8 cycloalkyl; and
(4) phenyl.
5. A compound of claim 4 wherein:
R.sup.7 and R.sup.8 are independently:
(1) hydrogen;
(2) C.sub.1-10 alkyl;
(3) C.sub.3-8 cycloalkyl;
(4) aminocarbonyl;
(5) substituted aminocarbonyl in which one or more substituent(s)
is:
(a) C.sub.1-5 alkyl,
(b) C.sub.3-8 cycloalkyl,
(c) phenyl,
(d) substituted phenyl in which the substituents are X and Y.
6. A compound of claim 5 wherein R.sup.1 is 1,1-dimethylpropyl or
sec-butyl.
7. A compound of claim 6 wherein R.sup.3 and R.sup.4 are
independently selected from:
(1) hydrogen;
(2) C.sub.1-5 alkyl; and
(3) phenyl.
8. A compound of claim 7 wherein a and c represent double
bonds.
9. A compound of claim 8 wherein R is ##STR54## 10.
10. A compound of claim 9 selected from the group consisting of:
.[.(1)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-hydroxymethyl-1,2,
6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2
H-pyran-2-one;
(2)
6(R)-[2-[8(S)-(2-methylbutyryloxy)-(S)-methyl-6(S)-hydroxymethyl-1,2,6,7,8
,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyr
an-2-one;
(3).]. .Iadd.(1)
.Iaddend.6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(R)-hydroxyme
thyl-1,2,6,7,8,8a-(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-te
trahydro-2H-pyran-2-one; .Iadd.and
(2)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-6(S)-phenylaminocarbonyloxymethyl-2
(S)-methyl-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5
,6-tetrahydro-2H-pyran-2-one .Iaddend.
.[.(4)
6(R)-[2-[B(S)-(2,2-dimethylbutyryloxy)-(S)-methyl-6(S)--phenylaminocarbony
loxymethyl-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5
,6-tetrahydro-2H-pyran-2-one.]..
11. A compound of claim 8 wherein R is CO.sub.2 R.sup.6 or
##STR55##
12. A compound of claim 11 selected from the group consisting of:
.[.(1)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-(S)-methyl-6(S)-carboxy-1,2,6,7,8,8
a(R)-hexahydro-naphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-H-pyran
-2-one;
(2)
6(R)-[2-[8(S)-(2-methylbutyryloxy)-2(S)-methyl-6(S)-carboxy-1,2,6,7,8,8a(R
)-hexahydronaphthyl-(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-H-pyran-2-on
e;
(3).]. .Iadd.(1)
.Iaddend.6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-.Iadd.2.Iaddend.(S)-methyl
-6(R)-carboxy-1,2,6,7,8,8-
a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-.Iadd.2
.Iaddend.H-pyran-2-one;
.[.(4).]. .Iadd.(2)
.Iaddend.6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(N,N.Iadd
.-.Iaddend.dimethyl)aminocarbonyl-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]et
hyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one;
.[.(5).]. .Iadd.(3)
.Iaddend.6(R)-[2-(8(S)-(2,2-dimethylbutyryloxy)-.Iadd.2.Iaddend.(S)-methyl
-6(R)-aminocarbonyl-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydr
oxy-3,4,5,6-tetrahydro-2H-pyran-2-one; .Iadd.and .Iaddend.
.[.(6).]. .Iadd.(4)
.Iaddend.6(R)-[2-(8(S)-(2-methylbutyryloxy)-2(S)-methyl-6(S)-aminocarbonyl
-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahy
dro-2H-pyran-2-one.[.; and the corresponding dihydroxy acids, and
esters thereof.]..
13. A compound of claim 9 selected from the group consisting
of:
(1)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(1-hydroxyethyl)
1
,2,6,7,8,8a-(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahyd
ro-2H-pyran-2-one;
(2)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(R)-(1-hydroxyethyl)
1
,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydr
o-2H-pyran-2-one;
(3) 6(R)-[2-[8(S)-(2
-methylbutyryloxy)-2(S)-methyl-6(S)-(1-hydroxyethyl)
1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahy
dro-2H-pyran-2-one;
(4)
6(R)-[2-[8(S)-(2-methylbutyryloxy)-2(S)-methyl-6(R)-(1-hydroxyethyl)-1,2,6
,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl,]4(R)-hydroxy-3,4,5,6-tetrahydro-2H
-pyran-2-one.
14. A compound of claim 7 wherein a, b and c represent single
bonds.
15. A compound of claim 14 wherein R is ##STR56##
16. A compound of claim 15 selected from the group consisting of:
(1)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-hydroxymethyl-1,2,
3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-
tetrahydro-2H-pyran-2-one;
(2)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-.Iadd.2.Iaddend.(S)-methyl-6(S)-(2,
2-dimethylbutyryloxymethyl)-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1
(S)]-ethyl]-4(R) -hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one;
(3)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-.Iadd.2.Iaddend.(S)-methyl-6(R)-(1-
hydroxyethyl)-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R
)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one;
(4)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-.Iadd.2.Iaddend.(S)-methyl-6(S)-(1-
hydroxyethyl)
1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4
,5,6-tetrahydro-2H-pyran-2-one;
(5) .[.6(R)-[2
(8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(1-hydroxyphenylmethyl)-1,
2,3,4,4a-(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5
,6-tetrahydro-2H-pyran-2-one.].6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-
methyl-6(S)-(1-hydroxyphenylmethyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronap
hthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one.
17. A compound of claim 14 wherein R is ##STR57##
18. A compound of claim 17 selected from the group consisting of:
(1)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-.Iadd.2(S)-methyl-.Iaddend.6(S)-car
boxy-1,2,3,4,4a(S),5,6,7,8,.Iadd.8.Iaddend.a(S)-decahydronaphthyl-1(S)]ethy
l]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one;
(2)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-.Iadd.2.Iaddend.(S)-methyl-6(S)-eth
oxycarbonyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-
hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one;
(3)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-aminocarbonyl-1,2,
3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-(S)]ethyl]-4(R)-hydroxy-3,4,5,6-t
etrahydro-2H-pyran-2-one; .Iadd.and .Iaddend.
(4)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(N-cyclohexylamino
carbonyl,N-cyclohexyl)aminocarbonyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydrona
phthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one.[.;
and the corresponding ring opened dihydroxy acids, and esters
thereof.]..
19. A compound of claim 14 wherein R is ##STR58## 20.
20. A compound of claim 19 which is: .[.6(R)-[2
(8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(1 oxoethyl)
1,2,3,4,4a(S),5,6,7,8,a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,
5,6-tetrahydro-2H-pyran-2-one and the corresponding ring opened
dihydroxy acid, and esters thereof.].
.Iadd.6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(1-oxoethyl)
-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4
,5,6-tetrahydro-2H-pyran-2-one.Iaddend..
21. A compound of claim 7 wherein one of a, b and c represents a
double bond.
22. A compound of claim 21 wherein R is ##STR59##
23. A compound of claim 22 selected from the group consisting of:
(1)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(R)-hydroxymethyl-1,2,
3,4,6,7,8,8a-(R)-octahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahy
dro-2H-pyran-2-one;
(2)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(R)-(2,2-dimethylbutyr
yloxymethyl)-1,2,3,4,6,7,8,8a(R)-octahydronaphthyl-(S)]ethyl]-4(R)-hydroxy-
3,4,5,6-tetrahydro-2H-pyran-2-one;
(3) 6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(R)-(1
hydroxyethyl)
1.2.3,4,6,7,8,8a(R)-octahydronaphthyl-1(S)]ethyl]-4(R)-hydro
xy3,4,5,6-tetrahydro-2H-pyran-2-one.
24. A hypocholesterolemic, hypolipidemic pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a nontoxic
effective amount of a compound as defined in claim 1.
25. A method of inhibiting cholesterol biosynthesis comprising the
administration to a subject in need of such treatment a nontoxic
therapeutically effective amount of a compound of claim 1.
.Iadd.
26. The compound of claim 9 which is
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-hydroxymethyl-1,2,
3,4,4a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-
pyran-2-one. .Iaddend..Iadd.27. The compound of claim 9 which is
6(R)-[2-[8(S)-(2-methylbutyryloxy)-2(S)-methyl-6(S)-hydroxymethyl-1,2,6,7,
8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-py
ran-2-one. .Iaddend..Iadd.28. The compound of claim 11 which is
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-carboxy-1,2,6,7,8,
8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyra
n-2-one. .Iaddend..Iadd.29. The compound of claim 11 which is
6(R)-[2-[8(S)-(2-methylbutyryloxy)-2(S)-methyl-6(S)-carboxy--1,2,6,7,8,8a(
R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2
-one. .Iaddend.
Description
BACKGROUND OF THE INVENTION
Hypercholesterolemia is known to be one of the prime risk factors
for ischemic cardiovascular disease, such as arteriosclerosis. Bile
acid sequestrants have been used to treat this condition; they seem
to be moderately effective but they must be consumed in large
quantities, i.e. several grams at a time and they are not very
palatable.
MEVACOR.RTM. (lovastatin), now commercially available, is one of a
group of very active antihypercholesterolemic agents that functions
by limiting cholesterol biosynthesis by inhibiting the enzyme
HMG-CoA reductase. In addition to the natural fermentation
products, mevastatin and lovastatin, there are a variety of
semi-synthetic and totally synthetic analogs thereof.
The naturally occurring compounds and their semisynthetic analogs
have the following general structural formulae: ##STR2## wherein: Z
is hydrogen, C.sub.1-5 alkyl or C.sub.1-5 alkyl substituted with a
member of the group consisting of phenyl, dimethylamino, or
acetylamino; and
R.sub.1 is: ##STR3## wherein Q is ##STR4##
R.sub.3 is H or OH; and
R.sub.2 is hydrogen or methyl; and a, b, c, and d represent
optional double bonds, especially where b and d represent double
bonds or a, b, c, and d are all single bonds.
U.S. Pat. No. 4,517,373 discloses semisynthetic hydroxy containing
compounds represented by the above general formula wherein R.sub.1
is ##STR5##
U.S. Pat. No. 4,517,859 and U.S. Pat. No. 4,448,979 also disclose
semisynthetic hydroxy-containing compounds represented by the above
general formula wherein R.sub.1 is ##STR6##
These compounds are prepared by the action of certain
microorganisms on the corresponding non-hydroxylated substrates.
One such organism described in U.S. Pat. No. 4,537,859 is of the
genus Nocardia.
U.S. Pat. No. 4,376,863 discloses a fermentation product, isolated
after cultivation of a microorganism belonging to the genus
Aspergillus, which has a hydroxy-containing butyryloxy side chain
and is represented by the above general formula wherein R.sub.1 is
##STR7##
Japanese unexamined patent application No. J59-122,483-A discloses
a semisynthetic hydroxy-containing compound represented by the
above general formula wherein R.sub.1 is ##STR8##
SUMMARY OF THE INVENTION
This invention relates to novel compounds which are HMG-CoA
reductase inhibitors and are useful as antihypercholesterolemic
agents. Specifically, the compounds of this invention are analogs
of mevinolin and related compounds which possess a hydroxyalkyl
group, acyloxyalkyl or carbamoyloxyalkyl group of the structure
##STR9## a carboxy group, an alkoxycarbonyl group, a carbamoyl
group of structure ##STR10## or a ketone group of the structure
##STR11## substituted on the 6-position of the polyhydronaphthyl
moiety. Additionally, pharmaceutical compositions of these novel
compounds, as the sole therapeutically active ingredient and in
combination with bile acid sequestrants, are disclosed. Other
embodiments of this invention are methods of treating disease
conditions in which hypercholesterolemia is an etiological factor,
and processes for preparing the novel compounds.
DETAILED DESCRIPTION OF THE INVENTION
The specific HMG-CoA reductase inhibitors of this invention are the
compounds represented by the following general structural formulae
(I) and (II): ##STR12## wherein:
R is ##STR13##
R.sup.1 and R.sup.5 are independently:
(1) C.sub.1-10 alkyl;
(2) substituted C.sub.1-10 alkyl in which one or more
substituent(s) is:
(a) halogen,
(b) hydroxy,
(c) C.sub.1-10 alkoxy,
(d) C.sub.1-5 alkoxycarbonyl,
(e) C.sub.1-5 acyloxy,
(f) C.sub.3-8 cycloalkyl,
(g) phenyl,
(h) substituted phenyl in which the substituents are X and Y,
(i) C.sub.1-10 alkylS(O).sub.n in which n is 0 to 2,
(j) C.sub.3-8 cycloalkylS(O).sub.n,
(k) phenylS(O).sub.n,
(l) substituted phenylS(O).sub.n in which the substituents are X
and Y, and
(m) oxo;
(3) C.sub.1-10 alkoxy;
(4) C.sub.2-10 alkenyl;
(5) C.sub.3-8 cycloalkyl;
(6) substituted C.sub.3-8 cycloalkyl in which one substituent
is:
(a) C.sub.1-10 alkyl,
(b) substituted C.sub.1-10 alkyl in which the substituent is:
(i) halogen,
(ii) hydroxy,
(iii) C.sub.1-10 alkoxy,
(iv) C.sub.1-5 alkoxycarbonyl,
(v) C.sub.1-5 acyloxy,
(vi) phenyl,
(vii) substituted phenyl in which the substituents are X and Y,
(viii) C.sub.1-10 alkylS(O).sub.n.
(ix) C.sub.3-8 cycloalkylS-(O).sub.n
(x) phenylS(O).sub.n,
(xi) substituted phenylS(O).sub.n in which the substituents are X
and Y, and
(xii) oxo,
(c) C.sub.1-10 alkylS(O).sub.n,
(d) C.sub.3-8 cycloalkylS(O).sub.n,
(e) phenylS(O).sub.n,
(f) substituted phenylS(O).sub.n in which the substituents are X
and Y,
(g) halogen,
(h) hydroxy,
(i) C.sub.1-10 alkoxy,
(j) C.sub.1-5 alkoxycarbonyl,
(k) C.sub.1-5 acyloxy,
(l) phenyl, and
(m) substituted phenyl in which the substituents are X and Y;
(7) phenyl;
(8) substituted phenyl in which the substituents are X and Y;
(9) amino;
(10) C.sub.1-5 alkylamino;
(11) di(C.sub.1-5 alkyl)amino;
(12) phenylamino;
(13) substituted phenylamino in which the substituents are X and
Y;
(14) phenyl C.sub.1-10 alkylamino;
(15) substituted phenyl C.sub.1-10 alkylamino in which the
substituents are X and Y;
(16) a member selected from:
(a) piperidinyl,
(b) pyrrolidinyl,
(c) piperazinyl,
(d) morpholinyl, and
(e) thiomorpholinyl; and
(17) R.sup.9 S in which R.sup.9 is:
(a) C.sub.1-10 alkyl,
(b) phenyl, and
(c) substituted phenyl in which the substituents are X and Y;
R.sup.2 and R.sup.6 are independently
(1) hydrogen;
(2) C.sub.1-5 alkyl;
(3) substituted C.sub.1-5 alkyl in which the substituent is
(a) phenyl,
(b) dimethylamino, and
(c) acetylamino, and
(4) 2,3-dihydroxypropyl;
R.sup.3 and R.sup.4 are independently:
(1) hydrogen;
(2) C.sub.1-10 alkyl;
(3) substituted C.sub.1-10 alkyl in which one or more
substituent(s) is
(a) halogen,
(b) hydroxy,
(c) C.sub.1-10 alkoxy,
(d) C.sub.1-5 alkoxycarbonyl,
(e) C.sub.1-5 acyloxy,
(f) C.sub.3-8 cycloalkyl,
(g) phenyl,
(h) substituted phenyl in which the substituents are X and Y,
(i) C.sub.1-10 alkylS(O).sub.n,
(j) C.sub.3-8 cycloalkylS(O).sub.n,
(k) phenylS(O).sub.n,
(l) substituted phenylS(O).sub.n in which the substituents are X
and Y, and
(m) oxo;
(4) C.sub.2-10 alkenyl;
(5) substituted C.sub.2-10 alkenyl in which one or more
substituent(s) is:
(a) halogen,
(b) hydroxy,
(c) C.sub.1-10 alkoxy,
(d) C.sub.1-5 alkoxycarbonyl,
(e) C.sub.1-5 acyloxy,
(f) C.sub.3-8 cycloalkyl,
(g) phenyl,
(h) substituted phenyl in which the substituents are X and Y,
(i) C.sub.1-10 alkylS(O).sub.n,
(j) C.sub.3-8 cycloalkylS(O).sub.n,
(k) phenylS(O).sub.n,
(l) substituted phenylS(O).sub.n in which the substituents are X
and Y, and
(m) oxo;
(6) C.sub.3-8 cycloalkyl;
(7) substituted C.sub.3-8 cycloalkyl in which one substituent
is:
(a) C.sub.1-10 alkyl,
(b) substituted C.sub.1-10 alkyl in which the substituent is:
(i) halogen,
(ii) hydroxy,
(iii) C.sub.1-10 alkoxy,
(iv) C.sub.1-5 alkoxycarbonyl,
(v) C.sub.1-5 acyloxy
(vi) phenyl,
(vii) substituted phenyl in which the substituents are X and Y,
(viii) C.sub.1-10 alkylS(O).sub.n,
(ix) C.sub.3-8 cycloalkylS(O).sub.n,
(x) phenylS(O).sub.n,
(xi) substituted phenylS(O).sub.n in which the substituents are X
and Y, and
(xii) oxo,
(c) C.sub.1-10 alkylS(O).sub.n,
(d) C.sub.3-8 cycloalkylS(O).sub.n,
(e) phenylS(O).sub.n,
(f) substituted phenylS(O).sub.n in which the substituents are X
and Y,
(g) halogen,
(h) hydroxy,
(i) C.sub.1-10 alkoxy,
(j) C.sub.1-5 alkoxycarbonyl,
(k) C.sub.1-5 acyloxy,
(l) phenyl, and
(m) substituted phenyl in which the substituents are X and Y;
(8) phenyl;
(9) substituted phenyl in which the substituents are X and Y;
R.sup.7 and R.sup.8 are independently:
(1) hydrogen;
(2) C.sub.1-10 alkyl;
(3) substituted C.sub.1-10 alkyl in which one or more
substituent(s) is
(a) halogen,
(b) hydroxy,
(c) C.sub.1-10 alkoxy,
(d) C.sub.1-10 alkoxycarbonyl,
(e) C.sub.1-5 acyloxy,
(f) C.sub.3-8 cycloalkyl,
(g) phenyl,
(h) substituted phenyl in which the substituents are X and Y,
(i) C.sub.1-10 alkyl S(O).sub.n in which n is 0 to 2,
(j) C.sub.3-8 cycloalkyl S(O).sub.n,
(k) phenyl S(O).sub.n ;
(l) substituted phenyl S(O).sub.n in which the substituents are X
and Y, and
(m) oxo;
(4) C.sub.2-10 alkenyl;
(5) C.sub.3-8 cycloalkyl;
(6) aminocarbonyl;
(7) substituted aminocarbonyl in which one or more substituent(s)
is
(a) C.sub.1-5 alkyl,
(b) C.sub.3-8 cycloalkyl,
(c) phenyl,
(d) substituted phenyl in which the substituents are X and Y;
(8) phenyl;
(9) substituted phenyl in which the substituents are X and Y;
(10) C.sub.1-10 alkylcarbonyl;
(11) C.sub.3-8 cycloalkylcarbonyl;
(12) phenylcarbonyl;
(13) substituted phenylcarbonyl in which the substituents are X and
Y; and
(14) a nitrogen-containing heterocyclic group selected from
piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl and
thiomorpholinyl; and
X and Y independently are hydrogen, halogen, trifluoromethyl,
C.sub.1-3 alkyl, nitro, cyano or a group selected from:
(1) R.sup.10 O(CH.sub.2).sub.m in which m is 0 to 3 and R.sup.10 is
hydrogen, C.sub.1-3 alkyl or hydroxy-C.sub.2-3 alkyl;
(2) ##STR14## in which R.sup.11 is hydrogen, C.sub.1-3 alkyl,
hydroxy-C.sub.2-3 alkyl, phenyl, naphthyl, amino-C.sub.1-3 alkyl,
C.sub.1-3 alkylamino-C.sub.1-3 alkyl, di(C.sub.1-3
alkyl)amino-C.sub.1-3 alkyl, hydroxy-C.sub.2-3 alkylamino-C.sub.1-3
alkyl or di(hydroxy-C.sub.2-3 alkyl) amino-C.sub.1-3 alkyl;
(3) ##STR15## in which R.sup.12 is hydrogen, C.sub.1-3 alkyl,
hydroxy-C.sub.2-3 alkyl, C.sub.1-3 alkoxy-C.sub.1-3 alkyl, phenyl
or naphthyl;
(4) ##STR16## in which R.sup.13 and R.sup.14 independently are
hydrogen, C.sub.1-3 alkyl, hydroxy-C.sub.2-3 alkyl or together with
the nitrogen atom to which they are attached form a heterocyclic
group selected from piperidinyl, pyrrolidinyl, piperazinyl,
morpholinyl or thiomorpholinyl;
(5) R.sup.15 S(O).sub.n (CH.sub.2).sub.m in which R.sup.15 is
hydrogen, C.sub.1-3 alkyl, amino, C.sub.1-3 alkylamino or
di(C.sub.1-3 alkyl)amino; and
a, b and c each represent single bonds or one of a, b and c
represents a double bond or both a and c represent double
bonds;
or a pharmaceutically acceptable salt thereof.
Except where specifically defined to the contrary, the terms
"alkyl", "alkoxy" and "acyl" include both the straight-chain and
branched-chain species of the term.
One embodiment of this invention is the class of compounds of the
formulae (I) and (II) wherein:
R.sup.1 and R.sup.5 are independently selected from:
(1) C.sub.1-10 alkyl;
(2) substituted C.sub.1-10 alkyl in which one or more
substituent(s) is:
(a) halogen,
(b) hydroxy,
(c) C.sub.1-10 alkoxy,
(d) C.sub.1-5 alkoxycarbonyl,
(e) C.sub.1-5 acyloxy,
(f) C.sub.3-8 cycloalkyl,
(g) phenyl,
(h) substituted phenyl in which the substituents are X and Y,
and
(i) oxo;
(3) C.sub.3-8 cycloalkyl;
(4) substituted C.sub.3-8 cycloalkyl in which one substituent
is:
(a) C.sub.1-10 alkyl,
(b) substituted C.sub.1-10 alkyl in which the substituent is
selected from
(i) halogen,
(ii) hydroxy,
(iii) C.sub.1-10 alkoxy
(iv) C.sub.1-5 acyloxy,
(v) C.sub.1-5 alkoxycarbonyl,
(vi) phenyl,
(vii) subsituted phenyl in which the substituents are X and Y,
and
(viii) oxo,
(c) halogen,
(d) hydroxy,
(e) C.sub.1-10 alkoxy,
(f) C.sub.1-5 alkoxycarbonyl,
(g) C.sub.1-5 acyloxy,
(h) phenyl,
(i) substituted phenyl in which the substituents are X and Y;
(5) phenylamino;
(6) substituted phenylamino in which the substituents are X and
Y;
(7) phenylC.sub.1-10 alkylamino; and
(8) substituted phenylC.sub.1-10 alkylamino in which the
substituents are X and Y.
One subclass of this embodiment is the compounds of formulae (I)
and (II) wherein:
R.sup.1 and R.sup.5 are independently:
(1) C.sub.1-10 alkyl;
(2) C.sub.3-8 cycloalkyl;
(3) phenylamino; and
(4) substituted phenylamino in which the substituents are X and
Y.
Illustrating this subclass are the compounds wherein R.sup.3 and
R.sup.4 are independently:
(1) hydrogen
(2) C.sub.1-10 alkyl;
(3) C.sub.3-8 cycloalkyl;
(4) phenyl.
Further illustrating this subclass are those compounds wherein
R.sup.7 and R.sup.8 are independently:
(1) hydrogen,
(2) C.sub.1-10 alkyl,
(3) C.sub.3-8 cycloalkyl,
(4) aminocarbonyl,
(5) substituted amino carbonyl in which one or more substituents
is
(a) C.sub.1-5 alkyl,
(b) C.sub.3-8 cycloalkyl,
(c) phenyl,
(d) substituted phenyl in which the substituents are X and Y.
More specifically illustrating this subclass are those compounds of
formula (I) and (II) wherein R.sup.1 is 1,1-dimethylpropyl or
sec-butyl and R.sup.3 and R.sup.4 are independently:
(1) hydrogen,
(2) C.sub.1-5 alkyl,
(3) phenyl.
Exemplifying this subclass are those compounds of the formulae (I)
and(II) wherein both a and c represent double bonds, especially the
following compounds:
(1)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl6(S)-(hydroxymethyl)-1,2
,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-
2H-pyran-2-one;
(2)
6(R)-[2-[8(S)-(2,2-dimethylbutyryl-oxy)-2(S)-methyl-6(R)-(hydroxymethyl)-1
,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydr
o-2H-pyran-2-one;
(3)
6(R)-[2-[8(S)-(2-methylbutyryloxy)-2(S)-methyl-6(S)-(hydroxymethyl)-1,2,6,
7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-
pyran-2-one;
(4)
6(R)-[2-[8(S)-(2-methylbutyryloxy)-2(S)-methyl-6(R)-(hydroxymethyl)-1,2,6,
7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-
pyran-2-one; and
(5)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-carboxy-1,2,6,7,8,
8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyra
n-2-one;
(6)
6(R)-[2-[8(S)-(2-methylbutyryloxy)-2(S)-methyl-6(S)-carboxy-1,2,6,7,8,8a(R
)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-
one;
(7)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(R)-carboxy-1,2,6,7,8,
8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyra
n-2-one;
(8)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(N,N-dimethyl)amin
ocarbonyl-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,
6-tetrahydro-2H-pyran-2-one;
(9)
6(R)-[2-(8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(R)-aminocarbonyl-1,2,
6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2
H-pyran-2-one;
(10)
6(R)-[2-(8(S)-(2-methylbutyryloxy)-2(S)-methyl-6(S)-aminocarbonyl-1,2,6,7,
8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-py
ran-2-one; and the corresponding dihydroxy acids, and esters
thereof.
Another exemplification of the subclass wherein a and c are both
double bonds are the following compounds.
(1)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(1-hydroxyethyl)-1
,2,6,7,8,8a-(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahyd
ro-2H-pyran-2-one;
(2)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(R)-(1-hydroxyethyl)-1
,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydr
o-2H-pyran-2-one;
(3)
6(R)-[2-[8(S)-(2-methylbutyryloxy)-2(S)-methyl-6(S)-(1-hydroxyethyl)-1,2,6
,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H
-pyran-2-one;
(4)
6(R)-[2-[8(S)-(2-methylbutyryloxy)-2(S)-methyl-6(R)-(1-hydroxyethyl)-1,2,6
,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H
-pyran-2-one; and the corresponding ring opened dihydroxy acids,
and esters thereof.
Another example of this subclass are those compounds of the
formulae (I) and (II) wherein a, b and c represent single bonds,
especially the following compounds:
(1)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-hydroxymethyl-1,2,
3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-
tetrahydro-2H-pyran-2-one;
(2)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(2,2-dimethylbutyr
yloxymethyl)-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)
-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one;
(3)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-carboxy-1,2,3,4,4a
(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrah
ydro-2H-pyran-2-one;
(4)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-ethoxycarbonyl-1,2
,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6
-tetrahydro-2H-pyran-2-one;
(5)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-aminocarbonyl-1,2,
3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-
tetrahydro-2H-pyran-2-one;
(6)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(N-cyclohexylamino
carbonyl,N-cyclohexyl)aminocarbonyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydrona
phthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one;
(7)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(R)-(1-hydroxyethyl)-1
,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5
,6-tetrahydro-2H-pyran-2-one;
(8)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(1-hydroxyethyl)-1
,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5
,6-tetrahydro-2H-pyran-2-one;
(9)
6(R)-[2-(8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(1-hydroxyphenylme
thyl)-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydrox
y-3,4,5,6-tetrahydro-2H-pyran-2-one;
(10)
6(R)-[2-(8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(1-oxoethyl)-1,2,3
,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-t
etrahydro-2H-pyran-2-one and the corresponding ring opened
dihydroxy acids, and esters thereof.
Another example of this subclass are those compounds of the
formulae (I) and (II) wherein one of a, b or c represents a double
bond, especially the following compounds:
(1)
6(R)-[2-[8(S)-(2,2-dimethylbutyryl-oxy)-2(S)-methyl-6(R)-hydroxymethyl-1,2
,3,4,6,7,8,8a(R)-octahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahy
dro-2H-pyran-2-one;
(2)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(R)-(2,2-dimethylbutyr
yloxymethyl)-1,2,3,4,6,7,8,8a(R)-octahydronaphthyl-(S)]ethyl]-4(R)-hydroxy-
3,4,5,6-tetrahydro-2H-pyran-2-one;
(3)
6(R)-[2-[8(S)-(2,2-dimethylbutyryl-oxy)-2(S)-methyl-6(R)-(1-hydroxyethyl)-
1,2,3,4,6,7,8,8a(R)-octahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetr
ahydro-2H-pyran-2-one; and
the corresponding ring opened dihydroxy acids, and esters
thereof.
The compounds of formulae (I) and (II) wherein a and c represent
double bonds are conveniently prepared from lovastatin or its
analogs having a 6-methyl group by one of the following three
microbiological methods:
(a) adding the substrate to a growing culture of Nocardia
autotrophica for a suitable incubation period followed by
isolation, and derivatization if desired;
(b) collecting a culture of the bioconverting microorganism and
contacting the collected cells with the substrate; or
(c) preparing a cell-free, enzyme-containing extract from the cells
of the bioconverting microorganism and contacting this extract with
the substrate.
Cultivation of the bioconverting microorganism of the genus
Nocardia can be carried out by conventional means in a conventional
culture medium containing nutrients well known for use with such
microorganisms. Thus, as is well known, such culture media contain
sources of assimilable carbon and of assimilable nitrogen and often
inorganic salts. Examples of sources of assimilable carbon include
glucose, sucrose, starch, glycerin, millet jelly, molasses and
soybean oil. Examples of sources of assimilable nitrogen include
soybean solids (including soybean meal and soybean flour), wheat
germ, meat extracts, peptone, corn steep liquor, dried yeast and
ammonium salts, such as ammonium sulphate. If required, inorganic
salts, such as sodium chloride, potassium chloride, calcium
carbonate or phosphates, may also be included. Also, if desired,
other additives capable of promoting the production of
hydroxylation enzymes may be employed in appropriate combinations.
The particular cultivation technique is not critical to the process
of the invention and any techniques conventionally used for the
cultivation of microorganisms may equally be employed with the
present invention. In general, of course, the techniques employed
will be chosen having regard to industrial efficiency. Thus, liquid
culture is generally preferred and the deep culture method is most
convenient from the industrial point of view.
Cultivation will normally be carried out under aerobic conditions
and at a temperature within the range from 20.degree. to 37.degree.
C., more preferably from 26.degree. to 28.degree. C.
Method (a) is carried out by adding the substrate to the culture
medium in the course of cultivation. The precise point during the
cultivation at which the starting compound is added will vary
depending upon the cultivation equipment, composition of the
medium, temperature of the culture medium and other factors, but it
is preferably at the time when the hydroxylation capacity of the
micro-organism begins to increase and this is usually 1 or 2 days
after beginning cultivation of the microorganism. The amount of the
substrate added is preferably from 0.01 to 5.0% by weight of the
medium, more preferably from 0.05 to 0.5%, e.g., from 0.05 to 0.1%
by weight. After addition of the substrate, cultivation is
continued aerobically, normally at a temperature within the ranges
proposed above. Cultivation is normally continued for a period of
from 1 to 2 days after addition of the substrate.
In method (b), cultivation of the microorganism is first carried
out under conditions such as to achieve its maximum hydroxylation
capacity; this capacity usually reaches a maximum between 4 and 5
days after beginning the cultivation, although this period is
variable, depending upon the nature and temperature of the medium,
the species of microorganism and other factors. The hydroxylation
capacity of the culture can be monitored by taking samples of the
culture at suitable intervals, determining the hydroxylation
capacity of the samples by contacting them with a substrate under
standard conditions and determining the quantity of product
obtained and plotting this capacity against time as a graph. When
the hydroxylation capacity has reached its maximum point,
cultivation is stopped and the microbial cells are collected. This
may be achieved by subjecting the culture to centrifugal
separation, filtration or similar known separation methods. The
whole cells of the cultivating microorganism thus collected,
preferably, are then washed with a suitable washing liquid, such as
physiological saline or an appropriate buffer solution.
Contact of the collected cells of the microorganism of the genus
Nocardia with the substrate is generally effected in an aqueous
medium, for example in a phosphate buffer solution at a pH value of
from 5 to 9. The reaction temperature is preferably within the
range from 20.degree. to 45.degree. C., more preferably from
25.degree. to 30.degree. C. The concentration of the substrate in
the reaction medium is preferably within the range from 0.01 to
5.0% by weight. The time allowed for the reaction is preferably
from 1 to 5 days, although this may vary depending upon the
concentration of the substrate in the reaction mixture, the
reaction temperature, the hydroxylation capacity of the
microorganism (which may, of course, vary from species to species
and will also, as explained above, depend upon the cultivation
time) and other factors.
The cell-free, enzyme-containing extract employed in method (c) may
be obtained by breaking down the whole cells of the microorganism
obtained as described in relation to method (b) by physical or
chemical means, for example by grinding or ultrasonic treatment to
provide a disintegrated cellular mass or by treatment with a
surface active agent or an enzyme to produce a cellular solution.
The resulting cell-free extract is then contacted with the
substrate under the same conditions as are described above in
relation to method (b).
The microorganism useful in the novel process of this invention is
of the genus Nocardia. Of particular importance are the known
strains of microorganism, Nocardia autotrophica, subspecies
canberrica, ATCC 35203 of the culture MA-6181 and subspecies
amethystina ATCC 35204 of the culture MA-6180 of the culture
collection of Merck & Co., Inc., Rahway, N.J. It should be
noted that the culture MA-6180 preferentially affords the
hexahydronaphthyl compounds (a and c are double bonds) of this
invention wherein R is CH.sub.2 OH, although the compounds wherein
R is CO.sub.2 H are also formed. Additionally, when the culture
MA6181 is utilized in the bioconversion reaction, the compounds of
the invention wherein R is CO.sub.2 H are preferentially formed,
although the compounds wherein R is CH.sub.2 OH are also prepared.
A sample of the culture designated ATCC 35203 and ATCC 35204 is
available in the permanent culture collection of the American Type
Culture Collection at 12301 Parklawn Drive, Rockville, Md.
20852.
A novel microorganism deposited in the culture collection of Merck
& Co., Inc. and designated MA-6455 may also be utilized in the
bioconversion reaction as described in copending patent application
Ser. No. 181,877 filed Apr. 15, 1988.
After completion of the conversion reaction by any of the above
methods, the desired compound can be directly isolated, separated
or purified by conventional means. For example, separation and
purification can be effected by filtering the reaction mixture,
extracting the resulting filtrate with a water-immiscible organic
solvent (such as ethyl acetate), distilling the solvent from the
extract, subjecting the resulting crude compound to column
chromatography, (for example on silica gel or alumina) and eluting
the column with an appropriate eluent, especially in an HPLC
apparatus.
The compound of formula (I) and (II) wherein a and c are double
bonds may also be prepared following the synthetic methodology in
copending patent applications Ser. No. 131,695 filed Feb. 24, 1988,
and Ser. No. 161,530, Ser. No. 161,579, Ser. No. 161,529 all filed
May 6, 1988. Lovastatin or a 8'-acyl analog is the the starting
material in each case.
The compounds of formula (I) wherein a and c represent double bonds
and R is CO.sub.2 H can be converted cleanly, and without
epimerization of the methine group to which R is appended, to the
corresponding primary alcohols wherein R is CH.sub.2 OH as
illustrated in the following synthetic pathway: ##STR17## Compound
(1) is converted to the corresponding triethylammonium salt (2) in
a suitable organic solvent, preferably methylene chloride at room
temperature. Without isolation but with cooling, preferably to
-70.degree. C., compound (2) is allowed to react with isobutyl
chloroformate to afford the mixed anhydride (3). The resulting,
cold solution of compound (3) is added to a cold, preferably
0.degree. C., solution of a suitable reducing agent, such as sodium
borohydride in a suitable organic solvent, such as ethanol, to
afford compound (4). Alternatively compound (1) can be directly
converted to compound (3) following the procedure in copending
patent application Ser. No. 065,223 filed June 22, 1987.
Compounds of formula (I) wherein a and c are double bonds and R is
R.sup.3 CHOH are conveniently prepared from the aldehyde (i)
##STR18## by the addition to the formyl group of a Grignard reagent
or other alkylatinq agent followed by the removal of the 3'-OH
group by the use of an acid such as HF. Aldehyde (i) is prepared as
described in copending patent application Ser. No. 161529 filed
Feb. 29, 1988.
The compounds of formulae (I) and (II) wherein a, b and c all
represent single bonds are conveniently prepared from lovastatin
via the following synthetic pathway: ##STR19##
The starting material, lovastatin (5) is readily available or may
be prepared according to the fermentation procedures disclosed in
U.S. Pat. No. 4,231,938. The compound of the formula (5) is
hydrolyzed under the conditions disclosed in U.S. Pat. No.
4,444,784 and then the 4-hydroxy function in the lactone moiety is
protected with a suitable protecting group, exemplified here as a
t-butyldimethylsilyl group, according to the procedure disclosed in
U.S. Pat. No. 4,444,784 to yield the compound (6). Compound (6) is
then hydroqenated under the analogous conditions disclosed in U.S.
Pat. No. 4,351,844 to afford the compound (7). Compound (7) is then
treated with nitrosyl chloride in the presence of a base, such as a
trialkylamine, specifically, trimethylamine, triethylamine,
pyridine, N,N-dimethylbenzylamine and the like, to afford the
compound (8). Compound (8) is then subjected to a
photorearrangement to give the compound (9). Compound (9) is heated
to reflux in a protic solvent such as isopropanol and the like to
yield the compound (10). Compound (10) is converted into compound
(11) by treatment with an alkali metal nitrite, such as sodium
nitrite or potassium nitrite, in an aqueous organic acid such as
acetic acid, propionic acid or the like. Compound (11), which is a
hemiacetal, is in equilibrium with the hydroxy aldehyde, compound
(12). This equilibrium mixture of compound (11) and compound (12)
is treated with a reducing agent, such as sodium borohydride, to
afford compound (13). Compound (13), is converted to compound (14)
by treatment with benzyl chloromethyl ether in the presence of a
suitable base such as a trialkylamine, preferably
diisopropylethylamine, in a suitable organic solvent such as
methylene chloride in the cold, preferably at about 0.degree. C.
Compound (14) is acylated under suitable conditions utilizing the
appropriate alkanoyl halide or alkanoic acid to afford compounds
(15). Removal of the benzyloxymethyl protecting group from
compounds (15) via catalytic hydrogenation under standard
conditions provides compounds (16). The latter are oxidized to
compounds (17) by standard methods, including the method of Swern
involving the use of oxalyl chloride and dimethyl sulfoxide in
methylene chloride followed by triethylamine. Compound (17) is then
reacted with an appropriately substituted Grignard reagent to
afford compound (18) which after removal of the trialkylsilyl ether
protecting group under standard conditions yields the compounds of
formula (I) wherein R is ##STR20##
The compounds of formula (I) wherein R is ##STR21## are
conveniently prepared from compound (18) by oxidizing the
hydroxyalkyl group to the ketone of the structure ##STR22##
followed by a second Grignard reaction and the removal of the
trialkylsilyl ether protecting group.
The compounds of the formula (I) wherein R is CO.sub.2 H are
conveniently prepared from the corresponding hydroxymethyl
containing compound under mild oxidation conditions. The compounds
of the formula (16) are treated with tris(triphenylphosphine)
ruthenium (II) chloride to afford the 6-formyl derivative which is
then treated with sodium chlorite and sulfamic acid to give the
desired products.
The compounds of the formulae (I) and (II) wherein a represents a
double bond are conveniently prepared from the compound of the
formula (6) by the hydrogenation procedure using Wilkinson's
catalyst as disclosed in U.S. Pat. No. 4,444,784 and subjecting the
resultant
6(R)-[2-[8(S)-hydroxy-2(S)6(R)-dimethyl-1,2,3,4,6,7,8,8a(R)-octahydronapht
hyl-1(S)]ethyl]-4(R)-(t-butyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-
2-one to the above noted reaction sequence. Alternatively a
hydrogenation procedure employing a homogenous rhodium catalyst in
an alcoholic solvent as described in Ser. No. 092,804 filed Sept.
3, 1987 may be used.
The compounds of the formulae (I) and (II) wherein b represents a
double bond are conveniently prepared using an analogous reaction
sequence but utilizing
6(R)-[2-[8(S)-hydroxy-2(S),6(S)-dimethyl-1,2,3,5,6,7,8,8a(R)-octahydronaph
thyl-1(S)]ethyl]-4(R)-(t-butyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran
-2-one in place of the compound of the formula (7). This starting
material may be prepared according to the procedures disclosed in
U.S. Pat. No. 4,444,784. Alternatively the starting material may be
prepared following the hydrogenation procedure detailed in Ser. No.
092,803 filed Sept. 3, 1987.
The compounds of the formulae (I) and (II) wherein c is a double
bond are conveniently prepared using an analogous reaction sequence
but utilizing
6(R)-[2-[8(S)-hydroxy-2(S),6(S)-dimethyl-1,2,4a(R),5,6,7,8,8a(S)-octahydro
naphthyl-1(S)]ethyl]-4(R)-(t-butyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-p
yran-2-one in place of the compound of the formula (7). This
starting material may be prepared from the natural fermentation
product prepared according to the procedures disclosed in U.S. Pat.
No. 4,294,846. Alternatively this starting material may be prepared
by the employment of a homogenous rhodium or iridium catalyst as
detailed in copending U.S. patent application Ser. No. 092,802
filed Sept. 3, 1987.
The compounds (17), which are prepared as illustrated in the
preceeding synthetic pathway, each have the CHO group appended to
the polyhydronaphthalene ring as a 6.alpha.-substituent. The
corresponding 6.beta.-epimers are conveniently prepared via the
following synthetic pathway: ##STR23##
Treatment of compounds (17) under the conditions (i.e.,
tetra-n-butylammonium fluoride in THF buffered with HOAc) used to
deblock the tert-butyldimethylsilyl ether serves both to deblock
this protecting group in compounds (17) and to epimerize the
resulting 6.alpha.-aldehydes (I9) to afford a mixture of epimeric
compounds (19) and (20), the ratio of the 6.beta.-epimer to the
6.alpha.-epimer being determined by the exact reaction conditions
used in each instance. After isolation by chromatography on a
suitable support such as silica gel and the like, compounds (20)
can be selectively protected on the lactone hydroxy group, as e.g.
a silyoxy ether, and then converted to compounds of this invention
by utilizing analogous reaction conditions as those employed with
the transformations of compound (17) and (18).
Epimerization can also be carried out employing aluminum isoproxide
and aluminum oxide to form the .beta.-hydroxymethyl compound
following the procedure in Ser. No. 161,529 filed Feb. 29,
1988.
Alternatively, the appropriate dihydro or tetrahydro derivative of
mevinolin may be subjected to one of the microbiological methods
described above to afford the compounds of the formulae (I) and
(II), wherein R is CH.sub.2 OH or CO.sub.2 H.
Where the product formed by the above described synthetic pathways
is not the desired form of that compound, then that product may be
subjected to one or more further reactions such as hydrolysis,
salification, esterification, acylation, ammonolysis or
lactonization by conventional methods, as described in more detail
hereafter.
The starting compound may be a free carboxylic acid, its
corresponding lactone or a salt (e.g., metal, amino acid or amine
salt) or ester (particularly alkyl ester) thereof.
Preferred metal salts are salts with alkali metals, such as sodium
or potassium, salts with alkaline earth metals, such as calcium, or
salts with other metals such as magnesium, aluminum, iron, zinc,
copper, nickel or cobalt, of which the alkali metal, alkaline earth
metal, magnesium and aluminum salts are preferred, the sodium,
calcium and aluminum salts being most preferred.
Preferred amino acids to form amino acid salts are basic amino
acids, such as arginine, lysine, histidine,
.alpha.,.beta.-diaminobutyric acid or ornithine.
Preferred amines to form amine salts include t-octylamine,
dibenzylamine, dichlorohexylamine, morpholine, alkyl esters of
D-phenylglycine and D-glucosamine. Also preferred is ammonia to
form the ammonium salt.
Esters are preferably the alkyl esters, such as the methyl, ethyl,
propyl, isopropyl, butyl, isobutyl or pentyl esters, of which the
methyl ester is preferred. However, other esters such as phenyl
C.sub.1-5 alkyl, dimethylamino-C.sub.1-5 alkyl, or
acetylamino-C.sub.1-5 alkyl may be employed if desired.
Of the starting materials, the alkali metal salts, e.g., the sodium
or potassium salts, are particularly preferred, the sodium salt
being most preferred as it has been found that this gives the best
conversion of the substrate into the desired product.
Where the product obtained by the processes of the present
invention is a salt of the carboxylic acid of formula (II), the
free carboxylic acid itself can be obtained by adjusting the pH of
the filtrate to a value of 4 or less, preferably to a value of from
3 to 4. Any organic acid or mineral acid may be employed, provided
that it has no adverse effect upon the desired compound. Examples
of the many acids which are suitable include trifluoroacetic acid,
acetic acid, hydrochloric acid and sulphuric acid. This carboxylic
acid may itself be the desired product or it may be, and preferably
is, subjected to subsequent reactions, as described below,
optionally after such treatments as extraction, washing and
lactonization.
Metal salts of the carboxylic acids of formula (II) may be obtained
by contacting a hydroxide, carbonate or similar reactive compound
of the chosen metal in an aqueous solvent with the carboxylic acid
of formula (II). The aqueous solvent employed is preferably water,
or it may be a mixture of water with an organic solvent, preferably
an alcohol (such as methanol or ethanol). a ketone (such as
acetone), an aliphatic hydrocarbon (such as hexane) or an
ester(such as ethyl acetate). It is preferred to use a mixture of a
hydrophilic organic solvent with water. Such reactions are normally
conducted at ambient temperature but they may, if desired, be
conducted with heating.
Amine salts of the carboxylic acids of formula (II) may be obtained
by contacting an amine in an aqueous solvent with the carboxylic
acid of formula (II). Suitable aqueous solvents include water and
mixtures of water with alcohols (such as methanol or ethanol),
ethers (such as tetrahydrofuran), nitriles (such as acetonitrile)
or ketones (such as acetone); it is preferred to use aqueous
acetone as the solvent for this reaction. The reaction is
preferably carried out at a temperature of ambient or below, more
preferably a temperature of from 5.degree. to 10.degree. C. The
reaction immediately goes to completion. Alternatively, a metal
salt of the carboxylic acid of formula (II) (which may have been
obtained as described above) can be dissolved in an aqueous
solvent, after which a mineral acid salt (for example the
hydrochloride) of the desired amine is added, employing the same
reaction conditions as when the amine itself is reacted with the
carboxylic acid of formula (II) and the desired product is then
obtained by a salt exchange reaction.
Amino acid salts of the carboxylic acids of formula (II) may be
obtained by contacting an amino acid in aqueous solution with the
carboxylic acid of formula (II). Suitable aqueous solvents include
water and mixtures of water with alcohols (such as methanol or
ethanol) or ethers (such as tetrahydrofuran).
Esters, preferably alkyl esters, of the carboxylic acids of formula
(II) may be obtained by contacting the carboxylic acid of formula
(II) with an appropriate alcohol, preferably in the presence of an
acid catalyst, for example a mineral acid (such as hydrochloric
acid or sulphuric acid), a Lewis acid (for example boron
trifluoride) or an ion exchange resin. The solvent employed for
this reaction is not critical, provided that it does not adversely
affect the reaction; suitable solvents include benzene, chloroform,
ethers and the like. Alternatively, the desired product may be
obtained by contacting the carboxylic acid of formula (II) with a
diazoalkane, in which the alkane moiety may be substituted or
unsubstituted. This reaction is usually effected by contacting the
acid with an ethereal solution of the diazoalkane. As a further
alternative, the ester may be obtained by contacting a metal salt
of the carboxylic acid of formula (II) with a halide, preferably an
alkyl halide, in a suitable solvent; preferred solvents include
dimethylformamide, tetrahydrofuran, dimethylsulfoxide and acetone.
All of the reactions for producing esters are preferably effected
at about ambient temperature, but, if required by the nature of the
reaction system, the reactions may be conducted with heating.
The compounds of formula (I) in which R is ##STR24## may be
prepared from the compounds of formula (I) in which R is ##STR25##
by the treatment of tert-butyldimethylsilyl in the presence of an
organic base to afford the compounds in which the hydroxy group of
the lactone of the compounds of formula (I) are protected as a
tert-butyldimethylsilyl ether. Subsequent acylation using the
appropriate acyl halide, anhydride, isocyanate or carbamoyl
chloride, under standard conditions. Desilylation under standard
conditions will afford the compounds of formula (I) in which R is
##STR26##
The compounds of formula (I) in which R is CO.sub.2 R.sup.6 may be
prepared from the compounds of formula (I) in which R is CO.sub.2 H
by treatment with a diazoalkane such as trimethylsilyldiazomethane
in a suitable organic solvent such as ether, hexane and the like.
Other methods for preparing these ester derivatives involve first
protecting the lactone hydroxyl group with a suitable protecting
group such as the tert-butyldimethylsilyl or tetrahydropyranyl
groups under standard conditions (vida supra) followed by
activation of the CO.sub.2 H group using standard procedures such
as conversion to the acid chloride with oxalyl chloride or
treatment with N,N-dicyclohexylcarbodiimide and subsequent reaction
with the appropriate alcohol R.sup.6 OH. Finally, removal of the
lactone hydroxyl protecting group will afford the compounds of
formula (I) in which R is CO.sub.2 R.sup.6.
The compouds of the formula (I) in which R is ##STR27## may be
prepared from the compounds of formula (I) in which R is CO.sub.2 H
by activation of the CO.sub.2 H group using standard procedures
such as treatment with carbonyl diimidazole or with isobutyl
chloroformate in such organic solvents such as methylene chloride,
THF and the like followed by treatment with R.sup.7 R.sup.8 NH
hydrochloride in the first instance and R.sup.7 R.sup.8 NH and a
suitable base such as triethylamine in the second instance.
Alternatively, the lactone hydroxyl group may be protected with a
suitable protecting group (vida supra), the CO.sub.2 H group
converted to the acid chloride and reacted with the desired amine
of the formula R.sup.7 R.sup.8 NH and the lactone hydroxyl
protecting group removed to afford the desired amide derivatives of
formula (I) in which R is ##STR28## Finally, the subclass of
compounds of formula (I) wherein R is ##STR29## and either R.sup.7
or R.sup.8 is alkyl or cycloalkyl and the other is N-(substituted)
aminocarbonyl is prepared by treating the requisite compound of
formula (I) in which R is CO.sub.2 H with a carbodiimide of the
formula R.sup.7 N--C--NR.sup.8 in the absence of extraneous
nucleophiles and allowing the intially formed imino anhydride to
undergo intramolecular rearrangement to the desired acylurea.
Lactones of the carboxylic acids of formula (I) may be obtained by
lactonizing the carboxylic acids of formula (II) under ordinary
conditions known to one skilled in the art.
The compounds of this invention are useful as
antihypercholesterolemic agents for the treatment of
arteriosclerosis, hyperlipidemia familial hypercholesterolemia and
like diseases in humans. They may be administered orally or
parenterally in the form of a capsule, a tablet, an injectable
preparation or the like. It is usually desirable to use the oral
route. Doses may be varied, depending on the age, severity, body
weight and other conditions of human patients but daily dosage for
adults is within a range of from about 2 mg to 2000 mg (preferably
10 to 100 mg) which may be given in two to four divided doses.
Higher doses may be favorably employed as required.
The compounds of this invention may also be coadministered with
pharmaceutically acceptable nontoxic cationic polymers capable of
binding bile acids in a non-reabsorbable form in the
gastrointestinal tract. Examples of such polymers include
cholestyramine, colestipol and
poly[methyl-(3-trimethylaminopropyl)imino-trimethylene dihalide].
The relative amounts of the compounds of this invention and these
polymers is between 1:100 and 1:15,000.
The intrinsic HMG-CoA reductase inhibition activity of the claimed
compounds is measured in the in vitro protocol published in J. Med.
Chem., 28, p. 47-358 (1985) and described below:
Isolation of HMG-CoA Reductase
Male Holtzman Sprague-Dawley rats (225-250 g) were kept on reversed
lighting and fed Purina rat chow containing 3% cholestyramine for 7
days preceding their sacrifice by CO.sub.2 asphyxiation. Livers
were removed 6 hours into the dark cycle and used immediately to
prepare microsomes. HMG-CoA reductase was solubilized from the
freshly prepared microsomes by the method of Heller and Shrewsbury
[J. Biol. Chem., 1976, 251, 3815]and purified through the second
ammonium sulfate precipitation step as described by Kleinsek et al.
[Proc. Natl. Acad. Sci. USA, 1977, 74, 1431]. The enzyme
preparation was tested for HMG-CoA reductase potency and diluted
with 100 mM phosphate buffer (pH 7.2) so that 100 .mu.1 of the
enzyme solution, when added to the assay control, gave a value of
50,000-60,000 dpm. The enzyme preparation was stored at -80.degree.
C.
HMG-CoA Reductase Inhibition Assay
The assay is essentially the procedure of Shefer et al. [J. Lipid
Res., 1972, 13, 402]. The complete assay medium contained the
following in a total volume of 0.8 ml: phosphate buffer, pH 7.2,
100 mM; MgCl.sub.2, 3 mM; NADP, 3 mM; glucose-6-phosphate, 10 mM;
glucose-6-phosphate dehydro-genase, 3 enzyme units; reduced
glutathione 50 mM; HMG-CoA (glutaryl-3-.sup.14 C, New England
Nuclear), 0.2 mM (0.1 .mu.Ci); and partially purified enzyme stock
solution, 100 .mu.L.
Test compounds or compactin, after first being converted to the
sodium salt of their dihydroxy acid form in situ by addition of 1N
NaOH (1 equivalent), were added to the assay system in 10-.mu.L
volumes at multiconcentration levels. After a 40-minute incubation
at 37.degree. C. with shaking and exposure to air, the reaction was
stopped by the addition of 0.4 mL of 8 N HCl. After an additional
30-minute incubation period at 37.degree. C. to ensure the complete
lactonization of mevalonic acid to mevalonolactone, 0.2 ml of the
mixture was added to an 0.5.times.5.0 cm column containing 100-200
mesh Bio-Rex 5, chloride form (Bio-Rad), wetted with distilled
water, as described by Alberts et al. [Proc. Natl. Acad. Sci.
U.S.A., 1980, 77, 3967]. The unreacted [.sup.14 C]HMG-CoA was
absorbed on the resin and the [.sup.14 C]mevalonolactone was eluted
with distilled water (2.times.1 ml) directly into 7-ml
scintillation vials. Five milliliters of Aquasol-2 (New England
Nuclear) was added to each vial, and radioactivity was measured in
a Packard Tri Carb Prias scintillation counter. IC.sub.50 values
were determined by plotting percentage inhibition against test
compound concentration and fitting a straight line to the resulting
data by using the least-squares method. For estimation of relative
inhibitory potencies, compactin was assigned a value of 100 and the
IC.sub.50 value of the test compound was compared with that of
compactin determined simultaneously.
Representative of the intrinsic HMG-CoA reductase inhibitory
activities of the claimed compounds are the relative potencies
tabulated below for a number of the claimed compounds.
TABLE I
__________________________________________________________________________
Compounds of the Formula (II) wherein R.sup.2 = H Relative a b c
AS* R R.sup.1 Potency.sup.1
__________________________________________________________________________
db -- db S CH.sub.2 OH 1,1-dimethylpropyl 189 db -- db S CO.sub.2 H
1,1-dimethylpropyl 99 db -- db R CO.sub.2 H 1,1-dimethylpropyl 94
db -- db S CO.sub.2 H sec-butyl 26 db -- db R CH.sub.2 OH sec-butyl
47 db -- db S CONMe.sub.2 1,1-dimethylpropyl 82 db -- db S
CONEt.sub.2 1,1-dimethylpropyl 65 db -- db S ##STR30##
1,1-dimethylpropyl 726 db -- -- R CH.sub.2 OH 1,1-dimethylpropyl
213 db -- -- R 2,2-dimethyl- 1,1-dimethylpropyl 33 butyryloxy-
methyl -- -- -- S CH.sub.2 OH 1,1-dimethylpropyl 86 -- -- -- S
CH.sub.2 OH cyclohexyl 33 -- -- -- S CO.sub.2 H 1,1-dimethylpropyl
79 -- -- -- S 2,2-dimethyl- 1,1-dimethylpropyl 100 butyryloxy-
methyl -- -- -- S CONMe.sub.2 1,1-dimethylpropyl 75 -- -- -- S
CONH.sub.2 1,1-dimethylpropyl 107 -- -- -- S CHOHPH
1,1-dimethylpropyl 220 -- -- -- S CHOHCH.sub.3 (ISOMER A)
1,1-dimethylpropyl 131 -- -- -- S CHOHCH.sub.3 (ISOMER B)
1,1-dimethylpropyl 113 -- -- -- R CHOHCH.sub.3 (ISOMER A)
1,1-dimethylpropyl 111 -- -- -- R CHOHCH.sub.3 (ISOMER B)
1,1-dimethylpropyl 104 -- -- -- S CO.sub.2 Et 1,1-dimethylpropyl
125 -- -- -- S CO.sub.2 i-Pr 1,1-dimethylpropyl 80 -- -- -- S
##STR31## 1,1-dimethylpropyl 100
__________________________________________________________________________
.sup.1 Relative to compactin arbitrarily assigned a value of 100
*AS = absolute sterochemistry of the methine moiety to which R is
appende db = double bond -- = single bond
Included within the scope of this invention is the method of
treating arteriosclerosis, familial hypercholesterolemia or
hyperlipidemia which comprises administering to a subject in need
of such treatment a nontoxic, therapeutically-effective amount of
the compounds of formulae (I) or (II) or pharmaceutical
compositions thereof.
The following examples illustrate the preparation of the compounds
of the formulae (I) and (II) and their incorporation into
pharmaceutical compositions and as such are not to be considered as
limiting the invention set forth in the claims appended hereto.
EXAMPLE 1
The following media are utilized in the bioconversion reactions
described below:
______________________________________ Grams per liter distilled
water ______________________________________ Medium A Yeast extract
4.0 Malt extract 10.0 Nutrient broth 4.0 Dextrose 4.0 pH 7.4 Medium
sterilized for 20 min. at 121.degree. C. Medium B Dextrose 10.0
Polypeptone 2.0 Meat extract 1.0 Corn steep liquor 3.0 pH 7.0
Medium sterilized for 20 min. at 121.degree. C.
______________________________________
I. Culture Conditions and Bioconversion
A lyophilized tube of Nocardia autotrophica subsp. canberrica ATCC
35204 (MA-6180) was used to inoculate 18.times.175 agar slants
(Medium A) which were incubated at 27.degree. C. for 7 days. The
slant culture was washed with 5 ml of sterile medium B and
transferred to a 250 ml flask containing 50 ml of sterile medium B.
This first stage seed was grown at 27.degree. C. on a 220 rpm
shaker and, after 24 hours, 2 ml was transferred to another flask
of sterile medium B.
Grown under the above conditions, the second seed was used to start
the bioconversion culture: 20 ml of the seed culture was placed in
400 ml of sterile medium B in a 2L flask. After the culture had
grown for 24 hours, 80 mg of the sodium salt of
7-[1,2,6,7,8,8a(R)-hexahydro-2(S),6(R)-dimethyl-8(S)-(2,2-dimethylbutyrylo
xy)-1(S)-naphthyl]-3(R),3(R)-dihydroxyheptanoic acid was added to
each flask. The incubation was continued for 28 hours or until no
7-[1,2,6,7,8,8a(R)-hexahydro-2(S),6(R)-dimethyl-8(S)-(2,2-dimethylbutyrylo
xy)-1(S)-naphthyl]-3(R),5(R)-dihydroxyheptanoic acid could be
detected by HPLC. The whole broth was clarified by centrifugation
followed by filtration through Whatman No. 2 filter paper.
II. HPLC Methods
Aliquots of whole broth could be analyzed for
7-[1,2,6,7,8,8a(R)-hexahydro-2(S),6(R)-dimethyl-8(S)-(2,2-dimethylbutyrylo
xy)-1(S)-naphthyl]-3(R),5(R)-dihydroxyheptanoic acid derivatives by
HPLC. Filtered broth could be injected directly (10 to 20 .mu.l) or
after dilution with methanol. The compounds were separated on
reversed phase columns utilizing a gradient of 35 to 45 percent
aqueous acetonitrile at flow rates ranging between 1 and 3 ml/min.
Addition of glacial acetic acid or H.sub.3 PO.sub.4 (0.1 ml/L
mobile phase) was required for the separation of the free acids.
Derivatives of
7-[l,2,6,7,8,8a(R)-hexahydro-2(S),6(R)-dimethyl-8(S)-(2,2-dimethylbutyrylo
xy)-1(S)-naphthyl]-3-(R),5(R)-dihydroxyheptanoic acid were detected
by monitoring the absorbance at 238 nm, as well as the absorbance
ratio of 238 nm/228 nm. The Waters HPLC system included a WISP
auto-injector, model 710B equipped with models 510 and 590 pumps, a
model 490 UV-visible detector, and the 840 data system. A number of
columns were used successfully for the separations, including the
following: Waters .mu. Bondapak-C18, Altex Ultrasphere-C18, Rainin
Microsorb-C18 and a Brownlee MPLC-C8.
III. Methyl
7-[1,2,6,7,8,8a(R)-hexahydro-6(S)-hydroxymethyl-2(S)-methyl-8(S)-(2,2-dime
thylbutyryloxy)-1(S)-naphthyl]-3(R),5(R)-dihydroxyheptanoate.
The whole broth of three X 400 ml culture broth was combined and
filtered through celite and Whatman No. 2 filter paper. The
filtrate was acidified to pH 5.0 with 25 percent H.sub.3 PO.sub.4
and then extracted with three, 700 ml-portions of ethyl acetate.
Following concentration under vacuum (25.degree. C.), the organic
solution was extracted with four volumes of 0.1% NaHCO.sub.3. The
bicarbonate solution was slowly adjusted to pH 4.5 with ethyl
acetate which was subsequently concentrated to 100 ml in vacuo. The
concentrate was combined with 150 ml of diethyl ether containing an
excess of CH.sub.2 N.sub.2 and stirred overnight for preparation of
the methyl ester derivatives. Evaporation of the ether was
performed under a stream of nitrogen and the remaining solution was
washed with 100 ml of phosphate buffer, pH 7.0. The organic phase
was taken to dryness in vacuo and the resulting residue was
dissolved in a minimum of isopropanol. Final purification of methyl
7-[1,2,6,7,8,8a(R)-hexahydro-6(S)-hydroxymethyl-2(S)-methyl-8(S)-(2,2-dime
thylbutyryloxy)-1(S)-naphthyl]-3(R),5(R)-dihydroxyheptanoate was
accomplished by HPLC utilizing a Waters .mu.Bondapak-C18 column
(1.times.30 cm). The mobile phase was 34 percent aqueous CH.sub.3 3
CN at 4 ml/min. Methyl
7-[1,2,6,7,8,8a(R)-hexahydro-6(S)-hydroxymethyl-2(S)-methyl-8(S)-(2,2-dime
thylbutyryloxy)-1(S)-naphthyl]-3(R),5(R)-dihydroxyheptanoate had a
retention time at 31 minutes. After evaporation of the solvent, the
sample was dried under vacuum for 24 hours to afford the title
compound which was identified by NMR. 1.sub.H nmr (CDCl.sub.3)
.delta. 0.83 (3H, t, J=7 Hz), 0.89 (3H, d, J=7Hz), 1.107 (3H, s),
1.111 (3H, s), 2.16 (H, m), 3.51 (H, d of d, J=5.5, 10.5 Hz), 3.61
(H, d of d, J=5.5, 10.5 Hz), 3.69 (3H,s), 3.77 (H, m), 4.22 (H, m)
5.36 (H, bs), 5.50 (H, bs), 5.80 (H, d of d, 6, 9.5 Hz), 6.00 (H,
d, J=9.5 Hz).
IV. Isolation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(R)-carboxy-2(S)-methyl-1,2,6,7,8,
8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyra
n-2-one and
6(R)-[2-(8(S)-(2,2-Dimethylbutyryloxy)-6(S)-carboxy-2(S)-methyl-1,2,6,7,8,
8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyra
n-2-one.
The whole broth (1200 ml) was clarified as before and then adjusted
to pH 3.5 with H.sub.3 PO.sub.4. The filtrate was loaded on a HP-20
column (3.times.50 cm) which had been equilibrated with water
containing 0.1 percent CH.sub.3 COOH. After washing the column with
1 L of water and 1 L of 25 percent CH.sub.3 CN, the products were
eluted with 600 ml of 50 percent CH.sub.3 CN. The acetonitrile was
removed under vacuum at 35.degree. C. The water was taken to pH 8.0
with NaOH and washed with two 500 ml portions of CH.sub.2 Cl.sub.2
which was discarded. After readjusting the pH to 3.5 with H.sub.3
PO.sub.4, the derivatives were first extracted into 1.8 L ethyl
acetate and then back-extracted into 1 L of 1 percent NaHCO.sub.3.
The bicarbonate solution was acidified to pH 5 with acetic acid and
loaded on a HP-20 column (1.5.times.50 cm). Once the column was
washed with 700 ml of H.sub.2 O followed by 700 ml of 30 percent
CH.sub.3 CN, the column was eluted with a gradient of 30 to 50
percent CH.sub.3 CN. The fractions were monitored by UV absorbance
(228, 238, 248 nm) and by HPLC. Crude
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-6(S)-carboxy-2(S)-methyl-1,2,6,7,8,
8a(R)-hexahydronaphthyl-1(S)]ethyl]4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran
-2-one was collected at about 40 percent CH.sub.3 CN.
After removing the solvent in vacuo, the resulting residue was
sonicated with 20 ml of toluene for 10 minutes, 3 .mu.l of CF.sub.3
COOH was added and the mixture was heated for 30 minutes at
70.degree. C. The toluene was removed under vacuum at 70.degree. C.
and the resulting residue was dissolved in 300 .mu.l of CH.sub.3
CN. The preceding procedure was employed to convert the derivative
of 7-[1,2,6,7,8,8a(R)-hexa- hydro-2(S),6(R)-dimethyl-(b
2,2-dimethylbutyryloxy)-1(S)-naphthyl]-3(R),5(R)-dihydroxyheptanoic
acid to its lactone form for ease of isolation. Final purification
was accomplished by HPLC using an Altex-C8 column (1.times.25 cm)
and a gradient of CH.sub.3 CN/CH.sub.3 OH/H.sub.2 O/CH.sub.3 COOH
(20/30/50/0.01 to 25/30/45/0.01) at 2.7 ml/min.
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(S)-carboxy-2(S)-methyl-1,2,6,7,8,
8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyra
n-2-one had a retention time of 30 to 31 minutes and was identified
by NMR. 1.sub.H nmr (CDCl.sub.3) .delta. 0.82 (3H, t, J=7.5 Hz),
0.88 (3H, d, J=7 Hz), 1.11 (6H, s), 1.53 (H, m), 2.60 (H, m) 2.72
(H, d of d, J=5, 18 Hz), 3.29 (H, m), 4.365 (H, m), 4.60 (H, m),
5.39 (H, bs), 5.62 (H, bs), 5.83 (H, d of d, J=6, 10 Hz), 6.00 (H,
d, J=10 Hz)
An alternate final purification involved fractionation by
preparative HPLC using a Vydac C-18 column and eluting with 0-60%
CH.sub.3 CN/0.170 phosphoric acid. Application of this purification
technique to a partially-purified mixture of acidic materials (200
mg) afforded fractions A containing a less polar, major component
and fractions B containing a more polar, minor component.
Concentration of fractions A in vacuo to remove the bulk of the
CH.sub.3 CN gave an aqueous mixture which was extracted with
chloroform. The organic extract was washed with saturated brine,
dried (Na.sub.2 SO.sub.4), filtered and evaporated in vacuo to
provide
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-carboxy-1,2,6,7,8,
8a(R)-hexanaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-o
ne as a colorless solid, mp 167.degree.-170.degree. C.; 1.sub.H nmr
(CD.sub.3 CN) .delta. 6.04 (H, d, J=9.8 Hz), 5.88 (H, d of d,
J=9.7, 6.0 Hz), 5.62 (H, m), 5.33 (H, m), 4.56 (H, m), 4.23 (H, m),
3.23 (H, m), 2.62 (H, d of d, J=17.4, 4.8 Hz), 2.44 (H, d of d of
d, J=17.5, 3.7, 1.6 Hz), 1.12 (6H, s), 0.90 (3H, d, J=7.1 Hz), 0.83
(3H, t, J=7.5 Hz). Recrystallization of this 6.beta.-carboxy isomer
from EtoAc-Hexane did not alter the mp. Furthermore, this
6.beta.-carboxy isomer mp 167.degree.-170.degree. C., could be
obtained directly from the partially-purified mixture of acidic
materials (vida supra) by crystallization from di-n-butyl
ether.
Anal. Calc'd for C.sub.25 H.sub.36 O.sub.7 : C, 66.94; H, 8.09.
Found: C, 66.66; H, 8.41.
From fractions B (vida supra) there was obtained the corresponding
6.alpha.-carboxy isomer
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2-(S)-methyl-6(R)-carboxy-1,2,6,7,8
,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyr
an-2one, as a colorless solid, mp 189.degree.-194.degree. C.; 1 H
nmr (CD.sub.3 CN) .delta. 6.06 (H, d, J=9 Hz), 5.88 (H, d of d,
J=9.5, 5.9 Hz), 5.71 (H, m), 5.24 (H, m), 4.51 (H, m), 4.21 (H, m),
3.20 (H, m), 2.70 (H, m), 2.62 (H, d of d, J=17.4, 4.8 Hz), 2.44
(H, m), 1.06 (H, s), 1.03 (3H, s), 0.89 (3H, d, J=7.0 Hz), 0.82
(3H, t, J=7.5 Hz).
Anal Calc'd for C.sub.25 H.sub.36 O.sub.7 : C, 66.94; H, 8.09.
Found: C, 66.70; H, 8.38.
In a similar fashion Nocardia autotrophica subsp. canberrica ATCC
35203 (MA6181) was utilized in the bioconversion reaction with the
sodium salt of
7-[1,2,6,7,8,8a(R)-hexahydro-2(S),6(R)-dimethyl-8(S)-(2,2-dimethylbutyr
yloxy)-1(S)-naphthyl]-3(R),5(R)-dihydroxyheptanoic acid to afford
the desired products.
Additionally, the sodium salt of
7-[1,2,6,7,8,8a(R)-hexahydro-2(S),6(R)-dimethyl-8(S)-(2-methylbutyryloxy)-
1(S)-naphthyl]-3(R),5(R)-dihydroxyheptanoic acid, the sodium salt
of ring opened mevinolin, was subjected to analogous bioconversion
reactions utilizing both N. autotrophic subsp. amethystina ATCC
35204 (MA6180) and N. autotrophic subsp. canberrica ATCC 35203
(MA6181) to predominantly afford
6(R)-[2-[8(S)-(2-methylbutyryloxy)-6(S)-carboxy-2(S)-methyl-1,2,6,7,8,8a(R
)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-
one and methyl
7-[1,2,6,7,8,8a(R)-hexahydro-6(S)-hydroxymethyl-2(S)-methyl-8(S)-(2-methyl
butyryloxy)-1(S)-naphthyl]-3(R),5(R)-dihydroxyheptanoate,
respectively.
EXAMPLE 2
Preparation of
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-hydroxymethyl-1,2,
3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-
tetrahydro-2H-pyran-2-one
(a)
6(R)-[2-[8(S)-Nitrosyloxy-2(S),6(S)-dimethyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-d
ecahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4.5.6-tetr
ahydro-2H-pyran-2-one (2a)
A stream of nitrosyl chloride gas was passed into a stirred
solution of 6(R)-[2-[8(S)-hydroxy-2(S),
6(S)-dimethyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydro-naphthyl-1(S)]ethyl]-4
(R)-(tert-butyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2-one
(800 mg, 1.82 mmol) in pyridine (14 ml) at 0.degree. C. until the
solution was saturated (brownish fumes filled the reaction flask).
The resulting mixture was stirred at 0.degree. C. for another 10
minutes, poured into cold water and extracted with diethyl ether.
The extract was washed successively with dilute HCl, water and 5%
NaHCO.sub.3, dried (MgSO.sub.4), filtered and concentrated in vacuo
to afford the title compound as a white solid; mp
92.degree.-4.degree. C.; 1.sub.H nmr (CDCl.sub.3) .delta. 0.86 (3H,
d, J=7 Hz), 0.89 (9H, s), 0.99 (3H, d, J=7 Hz), 2.55 (H, m of d,
J=18 Hz), 2.60 (H, d of d, J=18,4 Hz), 4.28 (H, m), 4.53 (H, m),
5.84 (H, m).
Anal Calc'd for C.sub.25 H.sub.45 NO.sub.5 Si: C, 64.20; H, 9.70;
N, 3.00. Found: C, 64.09; H, 10.00; N, 3.06.
(b)
6(R)-[2-[8(S)-Hydroxy-2(S)-methyl-6(S)-nitrosylmethyl-1,2,3,4,4a(S),5,6,7,
8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4
,5,6-tetrahydro-2H-pyran-2-one (2b)
Nitrogen gas was passed through a solution of compound 2a (870 mg,
1.82 mmol) in benzene (320 ml) for 25 minutes. This solution was
irradiated under N.sub.2 with a 450 watt Hanovia medium pressure
mercury lamp (pyrex filter) for 40 minutes at room temperature. The
reaction mixture was then concentrated in vacuo and the residue
applied to a silica gel column. Elution of the column with
methylene chloride:acetone (50:1; v:v) followed by elution with
methylene chloride:acetone: isopropanol (100:10:2; v:v:v) yielded
the desired product as a foamy oil; 1.sub.H nmr (CDCl.sub.3)
.delta. 0.83 (3H, d, J=7 Hz), 0.88 (9H, s) 4.10 (H, bs), 4.29 (H,
m), 4.64 (2H, d, J=8 Hz), 4.67 (H, m).
(c)
6(R)-[2-[8(S)-Hydroxy-2(S)-methyl-6(S)-hydroxyiminomethyl-1,2,3,4,4a(S),5,
6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)
-3,4,5,6-tetrahydro-2H-pyran-2-one (2c)
Compound 2b (288 mg, 0.616 mmol) was dissolved in isopropanol (15
ml) and heated at reflux for 2 hours. After cooling the reaction
mixture was concentrated in vacuo to leave a residue which afforded
the title compound as a gummy oil; 1.sub.H nmr (CDCl.sub.3) .delta.
0.86 (3H, d, J=7 Hz), 0.90 (9H, s) 2.33 (H, d, J=14 Hz), 2.78 (H,
m), 4.11 (H, m), 4,32 (H, m), 4.66 (H, m), 7,50 (H, d, J=6 Hz).
(d)
6(R)-[2-[8-Hydroxy-2(S)-methyl-6(S)-formyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-dec
ahydronaphthyl-1(S)]-ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4,5,6-tetra
hydro-2H-pyran-2-one (2d)
Sodium nitrite (477 mg 6.83 mmol) was added at 0.degree. C. in one
portion to a stirred solution of compound 2c (324 mg 0.683 mmol) in
acetic acid (14 ml) and water (7 ml). The resulting mixture was
stirred at 0.degree. C. for 10 minutes, warmed to room temperature
and stirred for 2.5 hours. The mixture was then diluted with water
and extracted with diethyl ether. This ethereal extract was washed
with water, 5% NaHCO.sub.3 (twice), dried and filtered. Evaporation
of the filtrate in vacuo afforded a brownish oily residue whose nmr
spectrum is consistent with the structure for compound 2d; 1.sub.H
nmr (CDCl.sub.3) .delta. 0.80 (3H, d, J=7 Hz), 0.88 (9H, s), 4,30
(H, m) 4.55 (2H, m).
(e)
6(R)-[2-[8(S)-Hydroxy-2(S)-methyl-6(S)-hydroxymethyl-1,2,3,4,4a(S),5,6,7,8
,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4,
5,6-tetrahydro-2H-pyran-2-one (2e)
Powdered sodium borohydride (40 mg, 1.05 mmol) was added at
0.degree. C. to a stirred solution of compound 2d (296 mg, 0.651
mmol) in 95% ethanol (15 ml) in one portion. The resulting mixture
was stirred at 0.degree. C. for 0.5 hours, then slowly treated with
a solution of aqueous (NH.sub.4).sub.2 SO.sub.4 (0.7 g in 15 ml of
H.sub.2 O). The resulting mixture was stirred at O.degree. C. for
0.5 hours, diluted with water (60 ml) and extracted with diethyl
ether. This extract was washed with water, 5% NaHCO.sub.3, dried,
filtered and evaporated to give a crude sample which was purified
by flash chromatography. Elution of the column with methylene
chloride:acetone:isopropanol (100:10:2; v:v:v) afforded the desired
product as a white solid; mp 124.degree.-7.degree. C.; 1.sub.H nmr
(CDCl.sub.3) .delta. 0.83 (3H, d, J=7Hz), 0.90 (9H, s), 3.73 (H, d
of d, J=11,6 Hz), 3.79 (H, d of d, J=11,6 Hz), 4.10 (H, bs) 4.31
(H, m), 4.70 (H, m).
Anal Calc'd for C.sub.25 H.sub.46 O.sub.5 Si C, 66.03; H, 10.20.
Found: C, 66.07; H, 10.38.
(f)
6(R)-[2-[8(S)-Hydroxy-2(S)-methyl-6(S)-(tert-butyldiphenylsilyloxymethyl)-
1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-l(S)]ethyl]-4(R)-(tert-butyld
imethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2-one (2f)
A solution of tert butyldiphenylsilyl chloride (140 mg, 0.50 mmol)
in dimethylformamide (1 ml) was added at 0.degree. C. to a stirred
solution of compound 2e (0.150 g, 0.33 mmol) and imidazole (115 mg,
1.7 mmol) in dimethylformamide (4 ml). The resulting mixture was
stirred at 0.degree. C. for 15 minutes and then warmed to room
temperature and stirred for 15 hours. The mixture was poured into
cold water and extracted with diethyl ether. This ethereal extract
was washed with dilute HCl and 5% NaHCO.sub.3, dried, filtered and
evaporated to leave crude product 2f which was purified by flash
chromatography on a silica gel column. Elution of the column with
methylene chloride:acetone (5:1; v:v) gave the desired product as a
gummy oil; 1.sub.H nmr (CDCl.sub.3) .delta. 0.84 (3H, d, J=7 Hz),
0.90 (9H, s), 1.09 (9H, s), 2.99 (H, d, J=6 Hz), 3.7-3.85 (2H, m)
4.02 (H, m), 4.30 (H, m) 4.67 (H, m) 7,3-7.5 (6H, m), 7.65-7.8 (4H,
m).
(g)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-(tert-butyldipheny
lsilyloxymethyl)-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-
4(R)-(tert-butyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2-one
(2g)
Lithium bromide powder (0.200 g, 2.30 mmol) was added at room
temperature under N.sub.2 in one portion to a stirred solution of
2,2-dimethylbutyryl chloride (0.150 g, 1.11 mmol) in pyridine (3.5
ml). The resulting mixture was stirred at room temperature until it
became a homogenous solution (0.5 hours). 4-Dimethylaminopyridine
(DMAP) was added (80 mg, 0.65 mmol). To the resulting mixture was
added a solution of compound 2f (229 mg, 0.33 mmol) in pyridine
(2.5 ml). The resulting mixture as heated at 90.degree.-95.degree.
under N.sub.2 for 70 hours. The reaction mixture was cooled, poured
into cold water and extracted with diethyl ether. This ethereal
extract was washed successively with dilute HCl, water and 5%
NaHCO.sub.3, then dried, filtered and concentrated in vacuo to
afford an oily residue which was purified by flash chromatography
on silica gel, eluting with methylene chloride:acetone (200:1;
v:v). The product fractions were purified further by preparative
tlc (Analtech SiO.sub.2 plates, eluant=CH.sub.2 Cl.sub.2 :acetone
(75:1; v:v) to give the desired compound as a colorless viscous
oil; 1.sub.H nmr (CDCl.sub.3) .delta. 0.66 (3H, t, J=7 Hz), 0.84
(3H, d, J=7 Hz), 0.9 (9H, s). 0.91 (6H, s). 1.10 (9H, s) 3.51 (H, d
of d, J=11,4 Hz) 3.85 (H, t, J=11 Hz), 4.30 (H, m), 4.55 (H, m),
5.08 (H, m) 7.3-7.5(6H, m) 7.6-7.8 (4H, m).
(h)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-hydroxymethyl-1,2,
3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-l(S)]ethyl]-4(R)-hydroxy-3,4,5,6-
tetrahydro-2H-pyran-2one. (2h)
Tetra-n-butylammonium fluoride solution (1 ml, 1 M,1 mmol) was
added to a stirred mixture of compound 2g (55 mg, 0.0695 mmol) and
acetic acid (0.12 ml, 2.10 mmol) in tetrahydrofuran (1.2 ml). The
resulting mixture was stirred at room temperature for 36 hours. The
reaction mixture was heated at reflux for 4.5 hours, cooled to room
temperature and poured into cold water and extracted with diethyl
ether. The ethereal extract was washed with 5% NaHCO.sub.3, dried,
filtered, and concentrated in vacuo to yield a residue which was
purified by flash chromatography on silica gel. Elution of the
column with methylene chloride:acetone (10:1; v:v) removed the
impurities. Further elution with methylene
chloride:acetone:isopropanol (100:10:5; v:v:v) afforded the desired
compound as a gummy oil ; 1.sub.H nmr (CDCl.sub.3) .delta. 0.85
(3H, d, J=7 Hz), 0.87 (3H, t, J=7 Hz) 1.16 (3H, s), 1.17 (3H, s),
2.62 (H, m of d, J=18 Hz), 2.73 (H, d of d, J=18, 5 Hz), 3.0 (H,
bs), 3.57 (H, d of d, J=11,6 Hz), 3.80 (H, t, J=11 Hz) 4.34 (H, m),
4.60 (H, m) 5.20 (H, m).
Anal Calc'd for C.sub.25 H.sub.42 O.sub.6 : C, 68.46; H, 9.65.
Found: C, 68.35; H, 9.85.
EXAMPLE 3
Preparation of 6(R)-[2-[8(S) (2,2-dimethylbutyryloxy)-
2-(S)-methyl-6(S)-(1
hydroxyethyl)-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(
R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one
(a)
6(R)-[2-[8(S)-Nitrosyloxy-2(S),6(S)-dimethyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-d
ecahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4,5,6-tetr
ahydro-2H-pyran-2-one (3a)
A stream of nitrosyl chloride gas was passed into a stirred
solution of
6(R)-[2-[8(S)-hydroxy-2(S)-6(S)-dimethyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decah
ydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4,5,6-tetrahyd
ro-2H-pyran-2-one (800 mg, 1.82 mmol) in pyridine (14 ml) at
0.degree. C. until the solution was saturated (brownish fumes
filled the reaction flask). The resulting mixture was stirred at
0.degree. C. for another 10 minutes, poured into cold water and
extracted with diethyl ether. The extract was washed successively
with dilute HCl, water and 5% NaHCO.sub.3, dried (MgSO.sub.4),
filtered and concentrated in vacuo to afford the title compound as
a white solid; mp 92.degree.-4.degree. C.; 1.sub.H nmr (CDCl.sub.3)
.delta. 0.86 (3H, d, J=7 Hz), 0.89 (9H, s), 0.99 (3H, d, J=7 Hz),
2.55 (H, m of d, J=18 Hz), 2.60 (H, d of d, J=18,4 Hz), 4.28 (H,
m), 4.53 (H, m), 5.84 (H, m).
Anal Calc'd for C.sub.25 H.sub.45 NO.sub.5 Si: C, 64.20; H, 9.70;
N, 3.00. Found: C, 64.09; H, 10.00; N, 3.06.
(b)
6(R)-[2-[8(S)-Hydroxy-2(S)-methyl-6(S)-nitrosylmethyl-1,2,3,4,4a(S),5,6,7,
8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4
,5,6-tetrahydro-2H-pyran-2-one (3b)
Nitrogen gas was passed through a solution of compound 3a (870 mg,
1.82 mmol) in benzene (320 ml) for 25 minutes. This solution was
irradiated under N.sub.2 with a 450 watt Hanovia medium pressure
mercury lamp (pyrex filter) for 40 minutes at room temperature. The
reaction mixture was then concentrated in vacuo and the residue
applied to a silica gel column. Elution of the column with
methylene chloride:acetone (50:1; v:v) followed by elution with
methylene chloride:acetone: isopropanol (100:10:2; v:v:v) yielded
the desired product as a foamy oil; 1.sub.H nmr (CDCl.sub.3)
.delta. 0.83 (3H, d, J=7 Hz), 0.88 (9H, s) 4.10 (H, bs), 4.29 (H,
m), 4.64 (2H, d, J=8 Hz), 4.67 (H, m).
(c)
6(R)-[2-[8(S)-Hydroxy-2(S)-methyl-6(S)-hydroxyiminomethyl-1,2,3,4,4a(S),5,
6,7,8,8a(S)
decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyl-dimethylsilyloxy)-3,4,5,6-t
etrahydro-2H-pyran-2 one (3c)
Compound 3b (288 mg, 0.616 mmol) was dissolved in isopropanol (15
ml) and heated at reflux for 2 hours. After cooling the reaction
mixture was concentrated in vacuo to leave a residue which afforded
the title compound as a gummy oil; 1.sub.H nmr (CDCl.sub.3) .delta.
0.86 (3H, d, J=7 Hz), 0.90 (9H, s) 2.33 (H, d, J=14 Hz), 2.78 (H,
m), 4.11 (H, m), 4,32 (H, m), 4.66 (H, m), 7,50 (H, d, J=6 Hz).
(d)
6(R)-[2-[8-Hydroxy-2(S)-methyl-6(S)-formyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-dec
ahydronaphthyl-1(S)]-ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4,5,6-tetra
hydro-2H-pyran-2-one (3d)
Sodium nitrite (477 mg 6.83 mmol) was added at 0.degree. C. in one
portion to a stirred solution of compound 3c (324 mg 0.683 mmol) in
acetic acid (14 ml) and water (7 ml). The resulting mixture was
stirred at 0.degree. C. for 10 minutes, warmed to room temperature
and stirred for 2.5 hours. The mixture was then diluted with water
and extracted with diethyl ether. This ethereal extract was washed
with water, 5% NaHCO.sub.3 (twice), dried and filtered. Evaporation
of the filtrate in vacuo afforded a brownish oily residue whose nmr
spectrum is consistent with the structure for compound 2d; 1.sub.H
nmr CDCl.sub.3) .delta. 0.80 (3H, d, J=7 Hz), 0.88 (9H, s), 4,30
(H, m) 4.55 (2H, m).
(e)
6(R)-[2-[8(S)-Hydroxy-2(S)-methyl-6(S)-hydroxymethyl-1,2,3,4,4a(S),5,6,7,8
,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4,
5,6-tetrahydro-2H-pyran-2-one (3e)
Powdered sodium borohydride (40 mg, 1.05 mmol) was added at
0.degree. C. to a stirred solution of compound 3d (296 mg, 0.651
mmol) in 95% ethanol (15 ml) in one portion. The resulting mixture
was stirred at 0.degree. C. for 0.5 hours, then slowly treated with
a solution of aqueous (NH.sub.4).sub.2 SO.sub.4 (0.7 g in 15 ml of
2 H.sub.2 O). The resulting mixture was stirred at O.degree. C. for
0.5 hours, diluted with water (60 ml) and extracted with diethyl
ether. This extract was washed with water, 5% NaHCO.sub.3, dried,
filtered and evaporated to give a crude sample which was purified
by flash chromatography. Elution of the column with methylene
chloride:acetone:isopropanol (100:10:2; v:v:v) afforded the desired
product as a white solid; mp 124.degree.-7.degree. C.; 1.sub.H nmr
(CDCl.sub.3) .delta. 0.83 (3H, d, J=7 Hz), 0.90 (9H, s), 3.73 (H, d
of d, J=11,6 Hz), 3.79 (H, d of d, J=11,6 Hz), 4.10 (H, bs) 4.31
(H, m), 4.70 (H, m).
Anal Calc'd for C.sub.25 H.sub.46 O.sub.5 Si C, 66.03; H, 10.20.
Found: C, 66.07; H, 10.38.
(f)
6(R)-[2-[8(S)-hydroxy-2(S)-methyl-6(S)-benzyloxymethoxymethyl-1,2,3,4,4a(S
),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]-ethyl]-4(R)-tert-butyldimethylsilyl
oxy)-3,4,5,6-tetrahydro-2H-pyran-2-one (3f)
To a stirred solution of compound 3e (9.7 g, 21.3 mmol) and
diisopropylethylamine (10 mL, 57.4 mmol) in CH.sub.2 Cl.sub.2 (25
mL) cooled to 0.degree. C. was added dropwise a solution of benzyl
chloromethyl ether (3.76 g, 24 mmol) in CH.sub.2 Cl.sub.2 (10 mL).
The resulting mixture was stirred at 0.degree. C. for 10 minutes,
allowed to warm to room temperature, stirred at room temperature
for 22 hours and then poured into ice water. The heterogeneous
mixture was extracted with diethyl ether. The organic phase was
separated, washed successively with dilute HCl, water, aqueous
NaHCO.sub.3 and water, dried (Na.sub.2 SO.sub.4), filtered and
evaporated in vacuo to afford a residue which was purified by flash
chromatography on silica gel. Elution with CH.sub.2 Cl.sub.2
acetone (50:1; v:v) removed the impurities. Continued elution with
CH.sub.2 Cl.sub.2 acetone (20:1; V:V) provided the title compound
as a viscous oil; 1.sub.H nmr (CDCl.sub.3) .delta. 0.82 (3H, d, J=7
Hz), 0.88 (9H, s), 2.6 (2H, m), 2.70 (H, d, J=6 Hz), 3.75 (2H, m),
4.0 (H, m), 4.28 (H, m), 4.60 (H, d, J=12 Hz), 4.62 (H, d, J=12
Hz), 4.66 (H, m), 4.78 (H, d, J=6 Hz), 4.81 (H, d, J=6 Hz), 7.3
(5H, m).
(g)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-benzyloxymethoxyme
thyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-b
utyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2-one (3g)
Powdered lithium bromide (6.4 g, 74 mmol) was added under N.sub.2
to a stirred mixture of 2,2-dimethylbutyryl chloride (4.97 g, 37
mmol) in pyridine (100 mL) and the resulting mixture was stirred
and warmed to 40.degree. C. until a clear solution was obtained. To
the resulting solution was added 4-dimethylaminopyridine (0.3 g,
2.45 mmol) and a solution of 3f (7.25 g, 13 mmol) in pyridine (30
mL). The resulting mixture was stirred and heated at 90.degree. C.
for 3.5 hours, cooled to room temperature, poured into ice water
and extracted with diethyl ether. The organic phase was separated,
washed with dilute HCl, aqueous NaHCO.sub.3 and saturated brine,
dried (Na.sub.2 SO.sub.4), filtered and evaporated in vacuo to give
an oily residue which was purified by flash chromatography on
silica gel. Elution with CH.sub.2 Cl.sub.2 -acetone (50:1; v:v)
afforded the title compound as a viscous oil; 1.sub.H nmr
(CDCl.sub.3) .delta. 0.82 (3H, d, J=7HZ), 0.83 (3H, t, J=7 Hz), 0
88 (9H, s), 1.14 (3H, s), 1.15 (3H, s), 2.67 (2H, m), 3.39 (H, d of
d, J=10, 6 Hz), 3.86 (H, t, J=10 Hz), 4.27 (H, m), 4.54 (H, d, J=16
Hz), 4.61 (H, d, J=16 Hz), 4.74 (2H, s), 5.13 (H, m), 7.32 (5H,
m).
(h)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6-(S)-hydroxymethyl-1,2
,3,4,4a(S),5,6,7,8,8a
(S)-decahydronaphthyl-(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4,5,6
-tetrahydro-2H-pyran-2-one (3h)
A mixture of compound 3g (6.1 g, 8.85 mmol), 10% Pd/C (0.5 g) and
acetic acid (3 drops) in isopropanol (200 mL) was hydrogenated in a
Paar apparatus for 4 hours. The resulting mixture was treated with
powdered NaHCO.sub.3 (1 g), stirred for 15 minutes and filtered.
The filtrate was evaporated in vacuo to provide a residue which was
dissolved in toluene (100 mL). The resulting solution was
evaporated in vacuo to provide a residue which again was dissolved
in toluene (100 mL). Evaporation of this solution in vacuo gave a
residue which crystallized from diethyl ether hexane to provide the
title compound as a colorless solid, mp 70.degree.-71.degree. C.;
'H nmr (CDCl.sub.3) .delta. 0.82 (3H, d, J=7 Hz), 0.83 (3H, t, J=7
Hz), 0.87 (9H, s), 1.15 (3H, s), 1.16 (3H, s), 2.66 (2H, m), 3.55
(H, m), 3.78 (H, m), 4.28 (H, m), 4.65 (H, m), 5.14 (H, m).
Anal. Cal'd for C31H56O6Si: C, 67.34; H, 10.21. Found: C, 67.21; H,
10.35.
6(R)-[2-[(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-formyl-1,2,3,4,4a(S)
,5,6,7,8,8a(S)-decahydronaphthyl-1(S)]-ethyl]-4(R)-(tert-butyldimethylsilyl
oxy)-3,4,5,6-tetrahydro-2H-pyran-2-one (3i)
To a stirred solution of oxalyl chloride (152 mg, 1.2 mmol) in
CH.sub.2 Cl.sub.2 (10 mL) cooled at -78.degree. C. was added
dimethylsulfoxide 156 mg, 2 mmol) via syringe under N.sub.2. The
resulting mixture was stirred at -78.degree. C. for 15 minutes and
treated with a solution of compound 3h (383 mg, 0.693 mmol) in
CH.sub.2 Cl.sub.2 (5 mL) added dropwise. The resulting mixture was
stirred at -78.degree. C., for 30 minutes, treated with
triethylamine (253 mg, 2.5 mmol), stirred for an additional 10
minutes at -78.degree. C. warmed to room temperature, poured into
ice water and extracted with diethyl ether. The organic extract was
washed with aqueous NaHCO.sub.3 and water, dried (Na.sub.2
SO.sub.4), filtered and evaporated in vacuo to provide the title
compound as a pale yellow oil; 1.sub.H nmr (CDCl.sub.3) .delta.
0.83 (3H, d, J=7 Hz), 0.83 (3H, t, J=7 Hz), 0.89 (9H, s), 1.10 (3H,
s), 1.12 (3H, s), 2.58 (2H, m), 4.28 (H, m), 4.55 (H, m), 5.20 (H,
m), 9.63 (H, s).
(j)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-(1-hydroxyethyl)-1
,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]-ethyl]-4(R)-(tert-butyld
imethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2-one (3j)
A solution of methylmagnesium bromide (3.0M in diethyl ether, 0.50
mL, 1.5 mmol) was added dropwise to a stirred solution of compound
3i (700 mg, 1.27 mmol) dissolved in dry diethyl ether (20 mL) at
-78.degree. C. under an argon atmosphere. The cooling bath was
removed and the reaction mixture was stirred with gradual warming
to room temperature over about a 2 hour period before quenching
with a saturated brine solution (10 mL). The reaction mixture was
distributed between diethyl ether (100 mL) and water (50 mL). The
diethyl ether layer was separated, dried, filtered and evaporated
to leave crude product which was purified by flash chromatography
on a silica gel column. Elution of the column with
chloroform:acetone (40:1/v:v) gave the two diastereomeric alcohols
as clear colorless glasses.
Isomer A: R.sub.f =0.17, diagnostic nmr peaks (CDCl.sub.3) .delta.
4.02 (H, m), 4.27 (H, m), 4.54 (H, m), 5.12 (H, m).
Isomer B: R.sub.f =0.07, diagnostic nmr peaks (CDCl.sub.3) .delta.
3.98 (H, m), 4.27 (H, m), 4.56 (H, m), 5.16 (H, m).
(k) 6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(1
hydroxyethyl)-1,2,3,4,4a,(S),5,6,7,8,8a(R)-decahydronaphthyl-1(S)]ethyl]-4
(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one (Isomer A)
n-Tetrabutylammonium fluoride solution (1M in THF, 1.65 mL, 1.65
mmol) was added to a stirred solution of compound 3j isomer A (180
mg, 310 .mu.mol) and acetic acid (180 .mu.L) in tetrahydrofuran (3
mL). The resulting solution was stirred at room temperature for 18
hours. Thin layer chromatography now showed only a trace of silyl
ether and was thus diluted with water (30 mL) and the crude product
was extracted into diethyl ether (100 mL). The etherial extract was
washed with 0.1N hydrochloric acid (100 mL), water (2.times.50 mL),
saturated sodium bicarbonate solution (50 mL), dried, filtered, and
evaporated to yield the crude alcohol which was purified by flash
chromatography on silica gel. Elution of the column with methylene
chloride:acetone (6:1/v:v) gave the desired diol as a clear
colorless glass: R.sub.f =0.46 chloroform:acetone (4:1/v:v),
diagnostic nmr peaks (CDCl.sub.3) 6 2.6 (H, md, J=18 Hz), 2.73 (H,
dd, J=5, 18 Hz), 4.03 (H, m), 4.35 (H,m), 4.56 (H,m), 5.15
(H,m).
Anal Calc'd for C.sub.26 H.sub.44 O.sub.6 : C, 68,99; H, 9.80.
Found: C, 69.07; H, 9.82.
(l)
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-(1-hydroxyethyl)-1
,2,3,4,4a,(S),5,6,7,8,8a(R)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,
5,6-tetrahydro-2H-pyran 2-one (Isomer B)
Aqueous hydrofluoric acid (49%, 0.42 mL) was added dropwise to a
stirred solution of compound 3j isomer B (200 mg, 0.34 mmol) in
acetonitrile (8 mL) at about 10.degree. C. The cooling bath was
removed and the clear colorless solution was stirred at room
temperature for 11/4 hour. The reaction mixture was distributed
between diethyl ether (100 mL) and saturated sodium bicarbonate
solution (50 mL). The ethereal layer was dried, filtered, and
evaporated to provide crude diol which was purified by flash
chromatography on silica gel. Elution of the column with methylene
chloride:acetone (6:1/v:v) gave the desired diol as a clear
colorless glass: R.sub.f =0.31 chloroform:acetone (4:1/v:v),
diagnostic nmr peaks (CDCl.sub.3) .delta. 2.6 (H, md, J=18 Hz),
2.73 (H, dd, J=5,18 Hz), 3.98 (H, m), 4.14 (H, m), 4.56 (H,m), 5.18
(H, m). MS (FAB)m/z 453 (MH.sup.+).
Anal Calc'd for C.sub.26 H.sub.44 O.sub.6 : C, 68.99; H, 9.80.
Found: C, 68.96; H, 10.04.
EXAMPLE 4
(j)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-(.alpha.-hydroxybe
nzyl)-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert
butyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran 2 one (4j)
Utilizing the general method described in Example 3 step j except
that the methylmagnesium bromide was replaced by phenylmagnesium
chloride the above titled compound was obtained as a single
diastereomer.
(k) 6(R)
[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-(.alpha.-hydroxybenzyl-
1,2,3,4,4a(S),5,6,7,8,8a-(S)-decahydronaphthyl-1(S)]ethyl]-4(R)
hydroxy-3,4,5,6 tetrahydro-2H-pyran 2 one (4k)
Utilizing the general method described in Example 3 step 1 except
that 1; isomer A was replaced by 4j, the above titled compound was
obtained as a colorless solid following recrystallization from
acetonitrile water, mp 196.degree.-196.5.degree. C., diagnostic nmr
peaks (CDCl.sub.3) .delta. 2.64 (H, md, J=18 Hz), 2.74 (H, dd, J=5,
18 Hz), 4.35 (H, m) 4.59 (H, m), 4.88 (H, dd, J=3, 10.8 Hz), 5.29
(H, d, J=3 Hz).
Anal. Calc'd for C.sub.31 H.sub.46 O.sub.6 : C, 72.34; H, 9.01
Found: C, 72.29; H, 9.29.
EXAMPLE 5
Preparation of
6(R)-[2-[8(S)-(Cyclohexylcarbonyloxy)-6(S)-hydroxymethyl-2(S)-methyl-1,2,3
,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-t
etrahydro-2H-pyran-2-one
(a)
6(R)-[2-[8(S)-Cyclohexylcarbonyloxy)-6(S)-(tert-butyldiphenylsilyloxymethy
l)-2(S)-methyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(
R)-(tert butyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2-one
(5a)
A solution of cyclohexylcarbonyl chloride (73 mg, 0.5 mmol) in
pyridine (2 ml) was added to a stirred mixture of
6(R)-[2-[8(S)-hydroxy-2(S)-methyl-6(S)-(tert-butyldiphenylsilyloxymethyl)-
1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyld
imethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2-one (116 mg, 0.167
mmol), the compound 2f, and 4 (dimethylamino)pyridine (24 mg, 0.2
mmol) in pyridine (2 ml). The resulting mixture was stirred at room
temperature under N.sub.2 for 3 hours and then heated at 65.degree.
C. for 1.5 hours. After cooling, the reaction mixture was poured
into cold water and extracted with diethyl ether. The ethereal
extract was washed successively with dilute hydrochloric acid,
water and 5% NaHCO.sub.3. After drying, it was filtered and
evaporated in vacuo to leave a residue which was purified by flash
chromatography. Elution of the column with methylene
chloride:acetone (100:1; V:V) yielded the desired compound as a
colorless gummy oil; 1.sub.H nmr (CDCl.sub.3) .delta. 0.83 (3H, d,
J=7 Hz), 0.90 (9H, s), 1.08 (9H, s), 3.54 (H, d of d, J=11, 6 Hz),
3.83 (H, t, J=11 Hz), 4.28 (H, m), 4.56 (H, m), 5.04 (H, m), 7.32
7.5 (6H, m), 7.6 7.8 (4H, m).
(b) 6(R)-[2-[8(S)-(Cyclohexylcarbonyloxy)
6(S)-hydroxymethyl-2(S)-methyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphth
yl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2 one
(5b)
By following the general procedure of Example 2, Step (h), but
using compound 5a in place of compound 2h, there was obtained the
desired product as a colorless, gummy oil; 1.sub.H nmr (CDCl.sub.3)
.delta. 0.86 (3H, d, J=7 Hz), 2.30 (H, m), 2.63 (H, m of d, J=18
Hz), 2.76 (H of d, J=18, 5 Hz), 3 58 (H, d of d, J=11, 6 Hz), 3.81
(H, t, J=11 Hz), 4 37 (H, m), 4.60 (H, m), 5.18 (H, m).
Anal Calc'd for C.sub.26 H.sub.42 O.sub.6 .multidot.0.4H.sub.2 O:
C, 68.21; H, 9.42. Found: C, 68.15; H, 9.53.
EXAMPLE 6
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(R)-hydroxymethyl-2(S)methyl-1,2,3
,4,6,7,8,8a(R)-octahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydr
o-2H-pyran-2-one
(a)
6(R)-[2-[8(S)-Hydroxy-2(S),6(R)-dimethyl-1,2,3,4,6,7,8,8a(R)-octahydronaph
thyl-1(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-py
ran-2-one
A solution (200 ml) of 50% toluene in absolute ethanol was
deoxygenated by bubbling N.sub.2 through it for 15 minutes.
Wilkinson's catalyst (2 g) was added to the solution and the
mixture reduced on the Paar hydrogenation apparatus at 50 psi
H.sub.2 for 90 minutes.
6(R)-[2-[8(S)-Hydroxy-2(S),6(R)-dimethyl-1,2,6,7,8,8a-(R)-hexahydronaphthy
l-1(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran
-2-one (4.0 g, 9.0 mmol.) was added and the solution hydrogenated
at 58 psi H.sub.2 for two days. The solvent was removed in vacuo
and the residue stirred with diethyl ether (500 ml) for 15 minutes
and then filtered. The filtrate was evaporated in vacuo to give a
brown solid which was dissolved in toluene (200 ml) containing
thiourea (2.6 The mixture was heated at 80.degree. C. for 2 hours
and then cooled to 0.degree. C. and filtered. The filtrate was
evaporated in vacuo and the solid residue chromatographed on a
7.times.18 cm column of silica gel. The column was eluted with 20%
ethyl acetate in hexane and 25 ml fractions were collected.
Fractions 54-90 were combined and evaporated in vacuo to yield the
title compound as a colorless solid. Crystallization of the solid
from aqueous CH.sub.3 CN provided an analytical sample as colorless
needles, mp 145.degree.-6.degree. C.; 1.sub.H nmr (CDCl.sub.3)
.delta. 0.070 (3H, s), 0.077 (3H, s), 0.88 (9H, s), 0.90 (3H, d,
J=7 Hz), 1,17 (3H, d, J =7 Hz), 2,58 (2H, m), 4,16 (H, m), 4.28 (H,
m), 4.66 (H, m), 5.41 (H, m).
Anal Calc'd for C.sub.25 H.sub.44 O.sub.45 : C, 68.76, H, 10.50.
Found: C, 68.72; H, 10.32.
(b)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(R)-hydroxymethyl-2(S)-methyl-1,2,
3,4,6,7,8,8a(R)-octahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahyd
ro-2H-pyran-2-one
By substituting an equimolar amount of the title compound in Step
(a) of this example for
6(R)-[2-[8(S)-hydroxy-2(S),6(S)-dimethyl-1,2,3,4,4a-5,6,7,8,8a(S)-decahydr
onaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4,5,6-tetrahydro-
2H-pyran-2-one in Step (a) of Example 2 and then following the
general procedures of Steps (a) through (h) of Example 2, there was
obtained a corresponding amount of the title compound as an
amorphorus solid; 1.sub.H nmr (CDCl.sub.3) .delta. 0.85 (3H, t,
J=Hz), 0.90 (3H, d, J=7 Hz), 1,14 3H, s), 1,16 (3H, s), 3.54 (H,
m), 3.65 (H, m), 4.37 (H, m), 4.59 (H, m), 5.35 (H, m), 5.47 (H,
m).
Anal Calc'd for C.sub.25 H.sub.40 O.sub.6 .multidot.0.5H.sub.2 O:
C, 67.38; H, 9.57. Found: C, 67.66; H, 9.28.
EXAMPLE 7
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-carboxy-1,2,3,4,4a
(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]4(R)-hydroxy
3,4,5,6-tetrahydro-2H-pyran-2-one (a)
6(R)-[2-[8(S)-2,2-Dimethylbutryloyxy)-2(S)-methyl-6(S)-formyl-1,2,3,4,4a(S)
,5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy
3,4,5,6-tetrahydro-2H-pyran 2-one (7a)
A mixture of compound 2h (100 mg, 0.228 mmol),
tris(triphenylphosphine) ruthenium (II) chloride (120 mg, 0.125
mmol) and sodium carbonate (40 mg) in benzene (7 ml) and methylene
chloride (2 ml) was stirred at ambient temperature under N.sub.2
for 24 hours. The reaction mixture was diluted with diethyl ether
(10 ml) and filtered through diatomaceous earth and silica gel
which was subsequently washed with methylene chloride. The combined
filtrate and washings were concentrated in vacuo to yield a crude
residue. The residue was purified by flash chromatography on silica
gel eluted with methylene chloride:acetone:isopropanol (100:10:2;
V:V:V) to afford the desired product as a gummy oil; 1.sub.H nmr
(CDCl.sub.3) .delta. 0.82 (3H, d, J=7 Hz), 0.83 (3H, t, J=7 Hz),
1.10 (3H,s), 1.12 (3H,s), 2.60 (H, m of d, J=18 Hz), 2.73 (H, d of
d, J=18,5 Hz), 4.37 (H,m), 4.57 (H,m), 5.23 (H,m), 9.64 (H, s)
(b)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-carboxy-1,2,3,4,4a
(S)5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]4(R)-hydroxy
3,4,5,6-tetrahydro- 2H-pyran-2-one (7b)
To a stirred mixture of compound 7a (14 mg, 0.032 mmol) and
sulfamic acid (4.4 mg, 0.045 mmol) in THF (1.5 ml) and water (0.5
ml) was added solid sodium chlorite (80% active, 5.6 mg, 0.05
mmol). The reaction mixture was stirred at ambient temperature for
45 minutes, poured into cold water (15 ml) and extracted with
diethyl ether and methylene chloride. The organic phase was
separated, dried (MgSO.sub.4) filtered and evaporated in vacuo to
give a crude residue. The residue was purified by flash
chromatography on silica gel eluted with methylene
chloride:acetone:isopropanol (100:10:5; v:v:v) to afford the
desired product as a gummy oil; 1.sub.H nmr (CDCl.sub.3) .delta.
0.83 (3H, d, J=7 Hz), 0.83 (3H, t, J=7 Hz), 1.08 (3H, s), 1.09 (3H,
s), 2.14 (H, m of d, J=13 Hz), 2.73 (H, d of d, J=18,5 Hz), 4.36
(H, m), 4.57 (H, m), 5.19 (H, m).
EXAMPLE 8
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-methoxycarbonyl-1,
2,3,4,4a(S)5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]4(R)-hydroxy-3,4,5,6-
tetrahydro-2H-pyran-2-one
To a solution of compound 7b (9.6 mg, 0.021 mmol) in isopropanol (2
ml) at ambient temperature was added a solution of
trimethylsilyl-diazomethane 10% in hexane, 0.3 ml. The reaction
mixture was stirred at ambient temperature for 18 hours and then
concentrated in vacuo to give a crude residue. The residue was
purified by flash chromatography on silica gel eluted with
methylene chloride:acetone:isopropanol 100:10:2; v:v:v) to afford
the desired product as a gummy oil; 1.sub.H nmr (CDCl.sub.3)
.delta. 0.83 (3H, d, J=7 Hz), 0.83 (3H, t, J=7 Hz), 1.09 (3H, s),
1.12 (3H,3), 1.52 (2H, t, J=7 Hz), 2.16 (H, m of d, J=13 Hz), 2.60
(H, m of d, J=18 Hz), 2.74 (H, d of d, J=18,5 Hz), 3.66 (3H,s),
4.37 (H, m), 4.57 (H, m), 5.18 (H, m).
EXAMPLE 9
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(R)-(2,2-dimethylbutyryloxymethyl)
-2(S)-methyl-1,2,3,4,6,7,8,8a(R)-octahydronaphthyl-1(S)]-ethyl]-4(R)-hydrox
y-3,4,5,6-tetrahydro-2H-pyran-2-one
To a solution of tetra-n-butylammonium fluoride (211 .mu.l, 1M in
tetrahydrofuran, 0.21 mmol), acetic acid (17 mg, 0.28 mmol) and
tetrahydrofuran (5 ml) was added
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-6(R)-(2,2-dimethylbutyryloxymethyl)
-2(S)-methyl-1,2,3,4,6,7,8,8a(R)-octahydronaphthyl-1(S)]-ethyl]-4(R)-(tert-
butyldimethylsilyloxy)-3,4,5,6-tetra-hydro-2H-pyran-2-one which was
isolated from the reaction of 2,2-dimethylbutyryl chloride and
6(R)-2-[8(S)-hydroxy-6(R)-(tert-butyldimethylsilyloxymethyl)-2(S)-methyl-1
,2,3,4,6,7,8,8a(R)-octahydronaphthyl-1(S)]ethyl]4(R)-(tert-butyldimethylsil
yloxy)-2,3,5,6-tetra-hydro-2H-pyran-2-one according to the general
procedure of Example 2, Step (g). The reaction mixture was stirred
at ambient temperature for 18 hours and then evaporated in vacuo to
give a crude residue. The residue was partitioned between diethyl
ether (50 ml) and water (10 ml). The aqueous phase was washed with
diethyl ether (2.times.50 ml). The combined organic phase and the
washings were washed with saturated sodium bicarbonate (5 ml) and
brine (2.times.25 ml), dried (MgSO.sub.4) and evaporated in vacuo
to yield a gummy residue. The residue was purified by flash
chromatography on silica gel eluted with 10 percent acetone in
methylene chloride (150 ml) and then 20 percent acetone in
methylene chloride to afford the desired product. This product was
further purified by preparative high pressure liquid chromatography
and after trituration with hexane gave a crystalline product; mp
119.degree.-121.degree. C.; 1.sub.H nmr (CDCl.sub.3) .delta. 0.84
(6H, m), 0.90 (3H, d, J=7 Hz), 1.14 (12H, s), 3,83 (1.sub.H d of d,
J=10,9 Hz), 4.19 (H, d of d, J=10,9 Hz), 4.37 (H, m), 4.59 (H, m),
5.36 (H, m), 5.40 (H, m), FAB MS 535 (M+H), 557 (M+Na).
EXAMPLE 10
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(S)-hydroxymethyl-2(S)-methyl-1,2,
6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2
H-pyran-2-one
To a stirred solution of
6(R)-[2-[8(S)-(2,2-dimethylbutylbutyryloxy)-6(S)-carboxy-2(S)-methyl-1,2,6
,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro
2H-pyran 2-one (102 mg, 0.23 mmol) in 4 .ANG. sieve-dried CH.sub.2
Cl.sub.2 (2.3 mL) was added triethylamine (32 .mu.l, 0.23 mmol).
The resulting mixture was cooled to -70.degree. C. and isobutyl
chloroformate (30 .mu.l, 0.23 mmol) was added over a 30-second
period with stirring. After stirring for 30 minutes at -70.degree.
C., the mixture was allowed to warm to 0.degree. C. over a 20
minute period. The resulting solution was added over a 30 second
period to a freshly prepared solution of NaBH.sub.4 (8.8 mg, 0.23
mmol) in EtOH (2 ml) with stirring at 0.degree. C. After 10
minutes, the cold mixture was partitioned between EtOAc (20 mL) and
0.1N HCl. The organic phase was separated, washed with water
(2.times.5 mL) and saturated brine (5 mL), dried (Na.sub.2
SO.sub.4), filtered and evaporated viscous oil (95 mg).
Chromatography of this oil on silica gel using 0 10% CH.sub.30 H in
CHCl.sub.3 as eluant afforded the title compound which was
identical by comparative tlc and 1.sub.H nmr spectral analysis to
an authentic sample isolated from a microbiological fermentation
broth of the sodium salt of
7-[1,2,6,7,8,8a(R)-hexahydro-2(S),6(R)-dimethyl-8(S)-(2,2-dimethylbutyrylo
xy)-1(S)naphthyl]-3(R),5(R)-dihydroxyheptanoic acid.
EXAMPLE 11
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(R)-hydroxymethyl-2(S)-methyl-1,2,
6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2
H-pyran-2-one
By substituting an equimolar amount of the 6(R)-carboxylic acid for
the 6(S)-carboxylic acid used in Example 10 and using the procedure
described therein, there was obtained a corresponding amount of the
title compound.
EXAMPLE 12
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(S)-(N,N-dimethyl)aminocarbonyl-2(
S)methyl-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6
-tetrahydro 2H-pyran 2-one
To a stirred solution of 6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)
6(S)-carboxy-2(S)-methyl-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R
)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one (100 mg) in CH.sub.2
Cl.sub.2 (1 mL) cooled to 0.degree. C. and maintained under N.sub.2
was added dropwise a solution of carbonyl diimidazole (39 mg) in
CH.sub.2 Cl.sub.2 (1 mL). After stirring for 1 hour at 0.degree.
C., the mixture was treated with dimethylamine hydrochloride (20
mg) and stirred for an additional 30 minutes. Then the mixture was
partitioned between EtOAc and 1N HCl. The organic phase was
separated, washed with aqueous NaHCO.sub.3 and saturated brine,
dried (Na.sub.2 SO.sub.4), filtered and evaporated in vacuo to
afford a residual oil. Chromatography of this oil on silica gel
using a gradient of 1 5% CH.sub.30 H in CH.sub.2 Cl.sub.2 as eluant
afforded the title compound as a solid, mp 148.degree.-160.degree.
C. after recrystallation from EtOAc-hexane.
Anal. Calc'd for C.sub.27 H.sub.41 N).sub.6 : C, 68.18; H, 8.69; N,
2.94. Found: C, 67.95; H, 8.97; N, 3.06.
EXAMPLE 13
Preparation of
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-6(S)-(N,N-diethyl)aminocarbonyl-2(S
)-methyl-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6
-tetrahydro-2H-pyran-2-one
To a stirred solution of
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-6(S)-carboxy-2(S)-methyl-1,2,6,7,8,
8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyra
n-2-one
To a stirred solution of
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-6(S)-carboxy-2(S)-methyl-1,2,6,7,8,
8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyra
n-2-one (100 mg) in CH.sub.2 Cl.sub.2 (1.5 mL) cooled to 0.degree.
C. and maintained under N.sub.2 was added triethylamine (31.3 ml ).
After 15 minutes, isobutyl chloroformate (29.4 ml) was added and,
after an additional 15 minutes, diethylamine (23.7 ml) was added.
The resulting mixture was stirred at 0.degree. C. for 30 minutes
and then washed with 1N HCl followed by aqueous NaHCO.sub.3. The
organic phase was separated, dried (Na.sub.2 SO.sub.4), filtered
and evaporated in vacuo to afford a residual oil. Chromatography of
this oil on silica gel using a gradient of 1-5% CH.sub.30 H in
CH.sub.2 Cl.sub.2 as eluant afforded the title compound as a solid,
mp 154.degree.-155.degree. C. after recrystallization from EtOAc
hexane.
Anal. Calc'd for C.sub.29 H.sub.45 NO.sub.6 : C, 69.15; H, 9.01; N,
2.78. Found: C, 68.85; H, 9.09; N, 2.63.
EXAMPLE 14
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(S)-propylaminocarbonyl-2(S)-methy
l-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrah
ydro-2H-pyran-2-one
By substituting an equimolar amount of n-propylamine for the
diethylamine used in Example 13 and using the procedure described
therein, there was obtained the title compound as a colorless
solid, mp 96.degree.-105.degree. C.
EXAMPLE 15
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(S)-benzylaminocarbonyl-2(S)-methy
l-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrah
ydro-2H-pyran-2-one
By substituting an equimolar amount of benzylamine for the
diethylamine used in Example 13 and using the procedure described
therein, there was obtained the title compound as a viscous
oil.
EXAMPLE 16
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(S)-(2-hydroxyethyl)aminocarbonyl-
2(S)-methyl-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,
5,6-tetrahydro-2H-pyran-2-one
By substituting an equimolar amount of 2-hydroxyethylamine for the
diethylamine used in Example 13 and using the procedure described
therein, there was obtained the title compound as a viscous
oil.
EXAMPLE 17
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-6(S)-phenylaminocarbonyloxymethyl-2
(S)-methyl-1,2,6,7,8,8a(R)-hexahydronaphthyl
(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one
To a stirred solution of 6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)
6(S)-hydroxymethyl-2(S)-methyl-1,2,6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethy
l]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one (80 mg, 0.184
mmol) in sieve dried pyridine (3 mL) was added phenyl isocyanate
(22 mg, 0.184 mmol). The resulting solution was stirred at room
temperature for 72 hours and then was evaporated in vacuo to
provide an oily residue. The residue was partitioned between
CHCl.sub.3 (125 mL) and 0.1N HCl (25 mL). The organic phase was
separated, washed with 0.1N HCl (2.times.25 ml) and saturated brine
(25 mL), dried (Na.sub.2 SO.sub.4), filtered and evaporated in
vacuo to give crude product. Chromatography of the crude product on
silica gel using CHCl.sub.3 -CH.sub.30 H (98:2, v:v) as eluant
afforded the title compound as a colorless gum which solidified
upon trituration with hexane, mp 115.degree.-119.degree. C.
Anal. Calc'd for C.sub.32 H.sub.43 NO.sub.7 : C,69.41; H, 7.83; N,
2.53. Found: C,69.51; H, 7.95; N, 2.67.
EXAMPLE 18
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-(N,N-dimethyl)amin
ocarbonyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]4(R)-hyd
roxy-3,4,5,6-tetrahydro-2H-pyran-2-one
(a)
6(R)-[2-[8(S)-hydroxy-2(S-methyl-6(S)-benzyloxymethoxymethyl-1,2,3,4,4a(S)
,5,6,7,8,8a(S)-decahydronaphthyl-1(S)]-ethyl]-4(R)-tert-butyl-dimethylsilyl
oxy) 3,4,5,6-tetrahydro-2H-pyran-2-one (18a)
To a stirred solution of compound 2c (9.7 g, 21.3 mmol) and
diisopropylethylamine (10 mL, 57.4 mmol) in CH.sub.2 Cl.sub.2 (25
mL) cooled to 0.degree. C. was added dropwise a solution of benzyl
chloromethyl ether (3.76 g, 24 mmol) in CH.sub.2 Cl.sub.2 (10 mL).
The resulting mixture was stirred at 0.degree. C. for 10 minutes,
allowed to warm to room temperature, stirred at room temperature
for 22 hours and then poured into ice water. The heterogeneous
mixture was extracted with diethyl ether. The organic phase was
separated, washed successively with dilute HCl, water, aqueous
NaHCO.sub.3 and water, dried (Na.sub.2 SO.sub.4), filtered and
evaporated in vacuo to afford a residue which was purified by flash
chromatography on silica gel. Elution with CH.sub.2 Cl.sub.2
acetone (50:1; v:v) removed the impurities. Continued elution with
CH.sub.2 Cl.sub.2 acetone (20:1: V:V) provided the title compound
as a viscous oil; 1.sub.H nmr (CDCl.sub.3) .delta. 0.82 (3H, d, J=7
Hz), 0.88 (9H, s), 2.6 (2H, m), 2.70 (H, d, J=6 Hz), 3.75 (2H, m),
4.0 (H, m), 4.28 (H, m), 4.60 (H, d, J=12 Hz), 4.62 (H, d, J=12
Hz), 4.66 (H, m), 4.78 (H, d, J=6 Hz), 4.81 (H, d, J=6 Hz), 7.3
(5H, m).
(b)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-benzyloxymethoxyme
thyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-b
utyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2-one (18b)
Powdered lithium bromide (6.4 g, 74 mmol) was added under N.sub.2
to a stirred mixture of 2,2-dimethylbutyryl chloride (4.97 g, 37
mmol) in pyridine (100 mL) and the resulting mixture was stirred
and warmed to 40.degree. C. until a clear solution was obtained. To
the resulting solution was added 4 dimethylaminopyridine (0.3 g,
2.45 mmol) and a solution of 18a (7.25 g, 13 mmol) in pyridine (30
mL). The resulting mixture was stirred and heated at 90.degree. C.
for 3.5 hours, cooled to room temperature, poured into ice water
and extracted with diethyl ether. The organic phase was separated,
washed with dilute HCl, aqueous NaHCO.sub.3 and saturated brine,
dried (Na.sub.2 SO.sub.4), filtered and evaporated in vacuo to give
an oily residue which was purified by flash chromatography on
silica gel. Elution with CH.sub.2 Cl.sub.2 acetone (50:1; v:v)
afforded the title compound as a viscous oil; 1.sub.H nmr
(CDCl.sub.3) .delta. 0.82 (3H, d, J=7 HZ), 0.83 (3H, t, J=7 Hz),
0.88 (9H, s), 1.14 (3H, s), 1.15 (3H, s), 2.67 (2H, m), 3.39 (H, d
of d, J=10, 6 Hz), 3.86 (H, t, J=10 Hz), 4.27 (H, m), 4.54 (H, d,
J=16 Hz), 4.61 (H, d, J=16 HZ), 4.74 (2H, S), 5.13 (H, m , 7.32
(5H, m).
(c)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-hydroxymethyl-1,2,
3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl
(S)]ethyl]-4(R)-(tert-butyldimethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2
-one (18c)
A mixture of compound 18b (6.1 g, 8.85 mmol), 10% Pd/C (0.5 g) and
acetic acid (3 drops) in isopropanol (200 mL) was hydrogenated in a
Paar apparatus for 4 hours. The resulting mixture was treated with
powdered NaHCO.sub.3 (1 g), stirred for 15 minutes and filtered.
The filtrate was evaporated in vacuo to provide a residue which was
dissolved in toluene (100 mL). The resulting solution was
evaporated in vacuo to provide a residue which again was dissolved
in toluene (100 mL). Evaporation of this solution in vacuo gave a
residue which crystallized from diethyl ether-hexane to provide the
title compound as a colorless solid, mp 70.degree.-71.degree. C.;
'H nmr (CDCl.sub.3) .delta. 0.82 (3H, d, J=7 Hz), 0.83 (3H, t, J=7
Hz), 0.87 (9H, s), 1.15 (3H, s), 1.16 (3H, s), 2.66 (2H, m), 3.55
(H, m), 3.78 (H, m), 4.28 (H, m), 4.65 (H, m), 5.14 (H, m).
Anal. Cal'd for C.sub.31 H.sub.56 O.sub.6 Si: C, 67.34; H, 10.21.
Found C, 67.21; H, 10.35.
(d)
6(R)-[2-[8(S)-(2,2-Dimethybutyryloxy)-2(S)-methyl-6(S)-formyl-1,2,3,4,4a(S
),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]-ethyl]-4(R)-(tert-butyldimethylsily
loxy) 3,4,5,6-tetrahydro-2H-pyran-one (18d)
To a stirred solution of oxalyl chloride (152 mg, 1.2 mmol) in
CH.sub.2 Cl.sub.2 (10 mL) cooled at -78.degree. C. was added
dimethylsulfoxide (156 mg, 2 mmol) via syringe under N.sub.2. The
resulting mixture was stirred at -78.degree. C. for 15 minutes and
treated with a solution of compound 18c (383 mg, 0.693 mmol) in
CH.sub.2 Cl.sub.2 (5 mL) added dropwise. The resulting mixture was
stirred at -78.degree. C., for 30 minutes, treated with
triethylamine (253 mg, 2.5 mmol), stirred for an additional 10
minutes at -78.degree. C. warmed to room temperature, poured into
ice water and extracted with diethyl ether. The organic extract was
washed with aqueous NaHCO.sub.3 and water, dried (Na.sub.2
SO.sub.4), filtered and evaporated in vacuo to provide the title
compound as a pale yellow oil; 1.sub.H nmr (CDCl.sub.3) .delta.
0.83 (3H, d, J=7 Hz), 0.83 (3H, t, J=7 Hz), 0.89 (9H, s), 1.10 (3H,
s), 1.12 (3H, s), 2.58 (2H, m), 4.28 (H, m), 4.55 (H, m), 5.20 (H,
m), 9.63 (H, s).
(e)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-carboxy-1,2,3,4,4a
(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimethylsil
yloxy) 3,4,5,6-tetrahydro-2H-pyran-2-one (18e)
To a stirred mixture of compound 18d (380 mg, 0.69 mmol) and
sulfamic acid (97 mg, 1 mmol) in THF water (5:1; v:v; 24 mL) cooled
at 0.degree. C. was added sodium chlorite (113 mg, 80% active, 1
mmol) in one portion. The resulting mixture was stirred at
0.degree. C. for 10 minutes, warmed to room temperature and stirred
for 2 hours. Then the mixture was poured into aqueous sodium
thiosulfate and extracted with diethyl ether. The organic extract
was washed with water, dried (Na.sub.2 SO.sub.4), filtered and
evaporated in vacuo to provide a residue which was purified by
flash chromatography on silica gel. Elution with CH.sub.2 Cl.sub.2
acetone (10:1; v:v) afforded the title compound as a colorless gum;
1.sub.H nmr (CDCl.sub.3) .delta. 0.83 (3H, t, J=7 Hz), 0.84 (3H, d,
J=7 Hz), 0.89 (9H, s), 1.10 (3H, s), 1.12 (3H, s), 2.15 (H, d, J=9
Hz), 2.60 (2H, m), 2.68 (2H, m) 4.30 (H, m), 4.58 (H, m), 5.68 (H,
m).
(f)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-chlorocarbonyl-1,2
,3,4,4a(S),5,6,7,8,a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimet
hylsilyloxy) 3,4,5,6-tetrahydro-2H-pyran 2-one (18f)
DMF (1 drop) was added to a magnetically stirred solution of
compound 18e (56 mg, 0.1 mmol) and oxalyl chloride (19 mL, 0.22
mmol) in benzene (2 mL) The resulting mixture was stirred at
ambient temperature for 2 hours to provide a heterogeneous mixture
which was decanted. Evaporation of the decantate in vacuo gave the
title compound as a pale yellow oil; 1.sub.H nmr (CDCl.sub.3
.delta. 0.072 (3H, s), 0.082 (3H, s), 0.88 (9H, s), 1.13 (3H, s),
1.16 (3H, s), 3.02 (H, m), 4.28 (H, m), 4.55 (H, m), 5.20 (H,
m).
(g) 6(R)-[2-[8(S)-2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-(N,N
dimethyl)aminocarbonyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]
ethyl]-4(R)-(tert-butyl dimethylsilyloxy) 3,4,5,6-tetrahydro-2H
pyran 2-one (18g)
Dimethylamine was bubbled slowly into a magnetically-stirred
solution of compound 18f (58 mg, 0.1 mmol) in diethyl ether (10 mL)
until the precipitation of dimethylammonium chloride ceased. Then
the mixture was stirred at ambient temperature for 18 hours and
filtered. Evaporation of the filtrate in vacuo provided a residual
oil which was chromatographed on silica gel (230-400 mesh,
3.times.15 cm). Elution with isopropanol-hexane (1:5; v:v) afforded
the title compound as a colorless oil; 1.sub.H nmr (CDCl.sub.3
.delta. 0.073 (3H, s), 0.084 (3H, s), 0.88 (9H, s), 1.09 (6H, s),
2.89 (3H, s), 2.95 (3H, s), 4.30 (H, m), 4.56 (H, m), 5.14 (H,
m).
(h) 6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-(N,N
dimethyl)aminocarbonyl-1,2,3,4,4a(S),5,6,7,8,8a(S)
decahydronaphthyl-1(S)]-ethyl-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-o
ne (18h)
To a stirred solution of compound 18g (78 mg, 0.13 mmol) and acetic
acid (30mL, 0.525 mmol) in THF (10 mL) was added
tetra-n-butylammonium fluoride (1M in THF, 394 mL, 0.394 mmol .
After stirring at ambient temperature for 18 hours, additional
tetra-n-butylammonium fluoride (394 mL) and acetic acid (30 mL)
were added to the mixture and stirring was continued for 24 hours.
Evaporation of the resulting solution in vacuo gave a residue which
was partitioned between ether (50 mL) and water (20 mL). After
separating the phases, the aqueous phase was extracted with diethyl
ether 50 mL . The diethyl ether extracts were combined, washed with
aqueous NaHCO.sub.3 and saturated brine, dried (MgSO.sub.4) and
evaporated in vacuo to provide a viscous oil which was
chromatographed on silica gel (230 400 mesh, 3.times.15 cm).
Elution with isopropanol-hexane (1:3; v:v) afforded the title
compound as a colorless solid, mp 186.degree.-187.degree. C. after
recrystallization from diethyl ether hexane; 1.sub.H nmr
(CDCl.sub.3) .delta. 1.11 (6H, s), 2.88 (3H, s), 2.96 (3H, s), 4.37
(H, m), 4.54 (H, m), 5.17 (H, m).
Anal. Calc'd for C.sub.27 H.sub.45 NO.sub.6 : C, 67.61; H, 9.46; N,
2.92 Found: C, 67.26; H, 9.64; N, 2.77.
EXAMPLE 19
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-aminocarbonyl-1,2,
3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-
tetrahydro-2H-pyran-2-one
(a)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-aminocarbonyl-1,2,
3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldimet
hylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2-one (19a)
By substituting an equimolar amount of ammonia for the
dimethylamine used in Step (g) of Example 18 and using the
procedure described therein, there was obtained the title compound
as a colorless oil; 1.sub.H nmr (CDCl.sub.3) .delta. 0.073 (3H, s),
0.082 (3H, s), 0.883 (9H, s), 1.14 (6H, s), 4.28 (H, m), 4.56 (H,
m), 5.08 (H, m), 5.28 (H, bs), 5.52 (H, bs).
(b)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-aminocarbonyl-1,2,
3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-
tetrahydro-2H-pyran-2-one (19b)
By substituting an equimolar amount of compound 19a for compound
18g used in Step (h) of Example 18 and using the procedure
described therein, there was obtained the title compound as a
colorless solid, mp 116.degree.-118.degree. C.; 1.sub.H nmr
(CDCl.sub.3) .delta. 0.82 (3H, d, J=7 Hz), 0.83 (3H, t, J=7 Hz),
1.14 (3H, s), 1.15 (3H, s), 4.36 (H, m), 4.56 (H, m), 5.10 (H, m),
5.22 (H, bs), 5.50 (H, bs).
Anal Calc'd for C.sub.25 H.sub.41 NO.sub.6 : C,66.49; H, 9.15; N,
3.10. Found: C,66.79; H, 9.35; N, 2.86.
EXAMPLE 20
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-ethoxycarbonyl-1,2
,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6
-tetrahydro-2H-pyran-2-one
(a)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-ethoxycarbonyl-1,2
,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyldime
thylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2-one (20a)
To a stirred solution of compound 18f (120 mg, 0.205 mmol) in
pyridine (5 mL) was added 4-dimethylaminopyridine (25 mg, 0.205
mmol) and EtOH (33 mg, 0.72 mmol). The resulting mixture was
stirred at ambient temperature for 4 days and then was partitioned
between cold 3N HCl and ether. The organic phase was separated,
washed with 3N HCl (until pH 4 was sustained in the aqueous phase),
washed with aqueous NaHCO.sub.3, dried (Na.sub.2 SO.sub.4) and
filtered. Evaporation of the filtrate in vacuo gave an oily resiue
which was purified by flash chromatography on silica gel. Elution
with CH.sub.2 Cl.sub.2 acetone (95:5; v:v) afforded the title
compound as a viscous oil; 1.sub.H nmr (CDCl.sub.3) .delta. 0.88
(9H, s), 1.08 (3H, s), 1.11 (3H, s), 2.14 (H, d, J=12 Hz), 2.5-2.7
(3H, m), 4.0 (H, m), 4.18 (H, m), 4.28 (H, m), 4.55 (H, m), 5.13
(H, m).
(b)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-ethoxycarbonyl-1,2
,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6
-tetrahydro-2H-pyran-2-one (20b)
To a stirred solution of compound 20a (38 mg, 0.055 mmol) and
acetic acid (26 ml) in THF (1 mL) was added tetra n butylammonium
fluoride (1M in THF, 320 mL, 0.32 mmol). The resulting mixture was
stirred at room temperature for 20 hours and then poured into cold
aqueous NaHCO.sub.3 and extracted with diethyl ether. The organic
phase was separated, dried (Na.sub.2 SO.sub.4), filtered and
evaporated in vacuo to yield the title compound as a viscous oil;
1.sub.H nmr (CDCl.sub.3) .delta. 0.82 (3H, d, J=7 Hz), 0.83 (3H, t,
J=7 Hz), 1.07 (3H, s), 1.11 (3H, s), 2.15 (H, d, J=12 Hz), 2.74 (H,
d of d, J=16, 4 Hz), 4.03 (H, m), 4.18 (H, m), 4.37 (H, m), 4.57
(H, m), 5.19 (H, m).
Anal Calc'd for C.sub.27 H.sub.44 O.sub.7 : C, 67.47; H, 9.23.
Found: C, 67.73; H, 9.43.
EXAMPLE 21
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-isopropoxycarbonyl
-1,2,3,4,4a(S),-5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,
4,5,6-tetrahydro-2H-pyran-2-one
(a)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-isopropoxycarbonyl
-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-(tert-butyl
dimethylsilyloxy)-3,4,5,6-tetrahydro-2H-pyran-2-one (21a)
To a stirred solution of compound 18e (140 mg, 0.247 mmol),
4-dimethylaminopyridine (14 mg, 0.112 mmol) and isopropanol (39 mg,
0.67 mmol) in CH.sub.2 Cl.sub.2 (300 .mu.L) cooled to 0.degree. C.
was added dropwise a solution of N,N'-dicyclohexylcarbodiimide (76
mg, 0.37 mmol) in CH.sub.2 Cl.sub.2 (500 .mu.L). The resulting
mixture was stirred and allowed to come to room temperature
overnight. After collecting the precipitated solid, the filtrate
was evaporated in vacuo to provide an oilY residue which was
purified by flash chromatography on silica gel. Elution with
CH.sub.2 Cl.sub.2 -acetone (98:2; v:v) afforded a mixture of the
title compound [1.sub.H nmr CDCl.sub.3) .delta. 0.88 (9H, s), 2.68
(2H, m), 4.28 (H, m), 4.55 (H, m), 4.95 (H, m), 5.13 (H, m)]and the
acylurea by-product (19 by-product).
(b)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-isopropoxycarbonyl
-1,2,3,4,4a(S),-5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,
4,5,6-tetrahydro-2H-pyran-2-one (19b)
To a stirred solution of the mixture (90 mg) of compounds 19a and
19 by-product and acetic acid (70 L) in THF (2.5 mL) was added
tetra-n-butylammonium fluoride (1M in THF, 860 .mu.L, 0.86 mmol).
The resulting mixture was stirred at room temPerature for 20 hours
and then poured into cold 5 aqueous NaHCO.sub.3 and extracted with
diethyl ether. The organic phase was separated, dried (Na.sub.2
SO.sub.4), filtered and evaporated in vacuo to leave an oily
residue which crystallized from diethyl ether-hexane to afford the
title compound as a solid, mp 160.degree.-161.degree. C; 1.sub.H
nmr (CDCl.sub.3) .delta. 0.82 (3H, d, J=7 Hz), 0.83 (3H, t, J=7
Hz), 1.10 (3H, s), 1.13 (3H, s), 1.20 (3H, d, J=6 Hz), 1.22 (3H, d,
J=6 Hz), 2.15 (H, d, J=12 Hz), 2.74 (H, d of d, J=18, 6 Hz), 4.35
(H, 4.55 (H, m), 4.95 (H, m , 5.17 (H, m).
Anal Calc'd for C.sub.28 H.sub.46 O.sub.7 : C, 67.98; H, 9.37.
Found: C, 68.08; H, 9.62.
EXAMPLE 22
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-(N-cyclohexylamino
carbonyl,N-cyclohexyl)aminocarbonyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydrona
phthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one
The title compound was isolated as the by product from Step (b) of
Example 21 and was purified by recrystallization from diethyl ether
hexane which afforded a solid, mp 137.degree.-138.degree. C.;
1.sub.H nmr (CDCl.sub.3) .delta. 1.08 (3H, s), 1.09 (3H, s), 2.60
(H, d of d, J=18, 5 Hz), 2.74 (H, d of d, J=18, 6 Hz), 2.88 (H, m),
3.63 (3H, m), 4.37 (H, m), 4.58 (H, m), 5.16 (H, m).
Anal. Calc'd for C.sub.38 H.sub.62 N.sub.2 O.sub.7 : C,69.26; H,
9.49;N, 4.25. Found: C,68.82; H, 9.70,N, 4.11.
EXAMPLE 23
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(R)-hydroxymethyl-1,2,
3,4,4a(S),5,6,7,8,8
a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2H-pyra
n-2-one
(a)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(R)-formyl-1,2,3,4,4a(
s),5,6,7,8,8a(s)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahy
dro-2H-pyran-2-one (23a)
To a stirred solution of compound 18d (300 mg, 0 54 mmol) and
acetic acid (0.35 mL) in THF (10 mL) was added tetra n
butylammonium fluoride (1M in THF, 4.3 mL, 4.3 mmol . The resulting
mixture was stirred at room temperature for 15 hours and then
poured into cold water and extracted with diethyl ether. The
organic phase was separated, washed with aqueous NaHCO.sub.3, dried
(Na.sub.2 SO.sub.4, filtered and evaporated in vacuo to provide a
residual oil which was purified by chromatography on silica gel.
Elution with CH.sub.2 Cl.sub.2 acetone (9:1; v:v) afforded an
epimeric mixture of the title compound [1.sub.H nmr (CDCl.sub.3) w
0.85 3H, d, J=7 Hz), 1.18 (3H, s), 5.28 H, m), 9.64 (H, d, J=1
Hz)]and the corresponding 6.alpha.-epimer [1.sub.H nmr (CDCl.sub.3)
.delta. 0.83 (3H, d, J=7 Hz), 1.10 (3H, s), 1.12 (3H, s), 5.24 (H,
m), 9.60 (H, d, J=1 Hz)],
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-formyl-1,2,3,4,4a(
S),5,6,7,8,8a(s)
deca-hydronaphthyl-1(s)]ethyl]-4(R)-hydroxy-3,4,5,6-tetra-hydro-2H-pyran-2
-one (21 epimer). The mixture of epimers 23a and 23 epimer could be
separated by chromatography or used as such in Step (b) below.
(b)
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(R)-hydroxymethyl-1,2,
3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-
tetrahydro-2H-pyran-2-one (23b)
By substituting an equimolar amount of the epimeric mixture of 23a
and 23 epimer from Step (a) above for compound 2d in Step (e) of
Example 2 and using the procedure described therein, there was
obtained an epimeric mixture of the title compound and compound 2g,
the latter being identical to an authentic sample of 2g prepared as
described in Example 2. Separation of this epimeric mixture into
pure compounds 23b and 2g could be accomplished by
chromatography.
EXAMPLE 24
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(R)-carboxy-1,2,3,4,4a
(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrah
ydro-2H-pyran-2-one
By substituting an equimolar amount of compound 23a for compound 7a
in Step (b) of Example 7 and using the procedure described therein,
there was obtained the title compound.
EXAMPLE 25
Preparation of
6(R)-[2-[8(S)-(2,2-Dimethylbutyryloxy)-2(S)-methyl-6(S)-phenylaminocarbony
loxymethyl-1,2,3,4,4a(S),5,6,7,8,8a(S)-decahydronaphthyl-1(S)]-ethyl]-4(R)-
hydroxy-3,4,5,6-tetrahydro-2H-pyran-2-one
By substituting an equimolar amount of compound 2h for the
6(R)-[2-[8(S)-(2,2-dimethylbutyryloxy)-2(S)-methyl-6(S)-hydroxymethyl-1,2,
6,7,8,8a(R)-hexahydronaphthyl-1(S)]ethyl]-4(R)-hydroxy-3,4,5,6-tetrahydro-2
H-pyran-2-one in Example 17 and using the procedure described
therein, there was obtained the title compound.
EXAMPLES 28-35
Utilizing the general procedure described in Example 8 and starting
with the 6-carboxy derivatives prepared according to the
bioconversion reactions of Example 1, the following compounds of
the formula (I) with the indicated absolute stereochemistry (AS) at
C-6 are prepared from the appropriate diazoalkane:
______________________________________ Compound Number a b c AS R
R.sup.1 ______________________________________ 28 db -- db S
CO.sub.2 Me 1,1-dimethyl- propyl 29 db -- db R CO.sub.2 Me
1,1-dimethyl- propyl 30 db -- db S CO.sub.2 iPr sec-butyl 31 db --
db R CO.sub.2 iPr sec-butyl
______________________________________
Similarly, starting with the 6-carboxy derivatives which are
prepared utilizing the general procedures of Examples 7 and 24, the
following compounds of formula (I) with the indicated absolute
stereochemistry (AS) at C-6 are also prepared:
______________________________________ Compound Number a b c AS R
R.sup.1 ______________________________________ 32 -- -- -- R
CO.sub.2 iPr 1,1-dimethyl- propyl 33 db -- -- S CO.sub.2 Me
1,1-dimethyl- propyl 34 -- db -- S CO.sub.2 Me 1,1-dimethyl- propyl
35 -- -- db R CO.sub.2 Me sec-butyl
______________________________________
EXAMPLES 36-42
The following compounds of the formula (I) with the indicated
absolute sterochemistry (AS) at C-6 and wherein R is ##STR32## are
prepared from the corresponding compounds wherein R is CH.sub.2 OH
and in which the 4-hydroxy function of the lactone moiety is
protected as a tetrahydropyranyl ether group by a standard
acylation reaction utilizing the appropriate acyl halide or
anhydride followed by the deprotection of the 4-hydroxy
function:
__________________________________________________________________________
Compound Number a b c AS R R.sup.1
__________________________________________________________________________
36 db -- db S ##STR33## sec-butyl 37 db -- db S ##STR34##
1,1-dimethylpropyl 38 -- -- -- S ##STR35## sec-butyl 39 -- -- -- S
##STR36## 1,1-dimethylpropyl 40 db -- -- R ##STR37##
1,1-dimethylpropyl 41 -- db -- S ##STR38## 1,1-dimethylpropyl 42 --
-- db S ##STR39## sec-butyl
__________________________________________________________________________
EXAMPLES 43-49
Using the general procedure described in Example 17 and starting
with the corresponding compounds wherein R is CH.sub.2 OH, the
following compounds of the formula (I) with the indicated absolute
stereochemistry (AS) at C-6, and wherein R is ##STR40## and R.sup.5
is NH.sub.2 or a substituted amino moiety, are prepared:
______________________________________ Com- pound Number a b c AS R
R.sup.1 ______________________________________ 43 db -- db R
##STR41## 1,1-methylpropyl 44 db -- db S ##STR42## sec-butyl 45 db
-- -- R ##STR43## 1,1-dimethylpropyl 46 -- db -- S ##STR44##
1,1-dimethylpropyl 47 -- -- db S ##STR45## sec-butyl 48 -- -- -- S
##STR46## 1,1-dimethylpropyl 49 -- -- -- R ##STR47## sec-butyl
______________________________________
EXAMPLES 50-57
Utilizing the general acylation procedures disclosed in co-pending
patent applications Ser. No. 859,524, 859,534, 859,535, all filed
May 5, 1986, the following compounds of the formula (I) with the
indicated absolute stereochemistry (AS) at C-6 wherein the R.sup.1
substituent is elaborated are prepared from the 6-carboxy,
6-alkoxycarbonyl, the protected 6-hydroxymethyl and the
6-acyloxymethyl derivatives.
______________________________________ Com- pound Number a b c AS R
R.sup.1 ______________________________________ 50 db -- db S
CH.sub.2 OH cyclohexyl- 51 db -- db R CH.sub.2 OH HOCH.sub.2 52 db
-- db S CO.sub.2 H CH.sub.3 COCH.sub.2 53 -- -- -- S CO.sub.2 H
cyclobutyl- 54 -- -- -- R CH.sub.2 OH HOCH.sub.2 CH.sub.2
C(CH.sub.3).sub.2 55 db -- -- R CH.sub.2 OH HO(CH.sub.2).sub.3
C(CH.sub.3).sub.2 56 -- db -- S CO.sub.2 CH.sub.3 CH.sub.3
COCH.sub.2 CH.sub.2 C(CH.sub. 3).sub.2 57 -- -- db S ##STR48##
cyclohexyl ______________________________________
EXAMPLE 58-62
Utilizing the methodology of Example 3 and the procedures disclosed
in copending patent applications Ser. No. 131695 filed Dec. 12,
1987 and Ser. Nos. 161530, 161529 all filed Feb. 29, 1988 and the
hydrogenation procedure in U.S. Pat. No. 4,444,784 the following
compounds of the formula (I) with the indicated stereochemistry
(AS) at C-6 are prepared.
______________________________________ Compound Number a b c AS R
R.sup.1 ______________________________________ 58 db -- db S
CH.sub.3 CHOH 1,1-methylpropyl 59 db -- db R CH.sub.3 CHOH
1,1-methylpropyl 60 db -- db S CH.sub.3 CHOH sec-butyl 61 db -- db
R CH.sub.3 CHOH sec-butyl 62 db -- -- R CH.sub.3 CHOH
1,1-methylpropyl ______________________________________
EXAMPLE 63
Preparation of Ammonium Salts of Compounds II
The lactone 2h (1.0 mmol) from Example 2 is dissolved with stirring
in 0.1N NaOH (1.1 mmol) at ambient temperature. The resulting
solution is cooled and acidified by the dropwise addition of 1N
HCl. The resulting mixture is extracted with diethyl ether and the
extract washed with brine and dried (MgSO.sub.4). The MgSO.sub.4 is
removed by filtration and the filtrate saturated with ammonia (gas
to give a gum which solidified to provide the ammonium salt.
EXAMPLE 64
Preparation of Alkali and Alkaline Earth Salts of Compounds II
To a solution of 42 mg of lactone 2h from Example 2 in 2 ml of
ethanol is added 1 ml of aqueous NaOH (1 equivalent). After one
hour at room temperature, the mixture is taken to dryness in vacuo
to yield the desired sodium salt.
In like manner, the potassium salt is prepared using one equivalent
of potassium hydroxide, and the calcium salt, using one equivalent
of CaO.
EXAMPLE 65
Preparation of Ethylenediamine Salts of Compounds II
To a solution of 0.50 g of the ammonium salt from Example 63 in 10
ml of methanol is added 75 ml of ethylenediamine. The methanol is
stripped off under vacuum to obtain the desired ethylenediamine
salt.
EXAMPLE 66
Preparation of Tris(hydroxymethyl)aminomethane Salts of Compounds
II
To a solution of 202 mg of the ammonium salt from Example 63 in 5
ml of methanol is added a solution of 60.5 mg of
tris(hydroxymethyl) aminomethane in 5 ml of methanol. The solvent
is removed in vacuo to afford the desired tris(hydroxy
methyl)aminomethane salt.
EXAMPLE 67
Preparation of L-Lysine Salts of Compounds II
A solution of 0 001 mole of L-lysine and 0.0011 mole of the
ammonium salt from Example 63 in ml of 85% ethanol is concentrated
to dryness in vacuo to give the desired L-lysine salt.
Similarly prepared are the L-arginine, L-ornithine, and
N-methylglucamine salts.
EXAMPLE 68
Preparation of Tetramethylammonium Salts of Compounds II
A mixture of 68 mg of ammonium salt from Example 63 in 2 ml of
methylene chloride and 0.08 ml of 24% tetramethylammonium hydroxide
in methanol is diluted with ether to yield the desired
tetramethylammonium salt.
EXAMPLE 69
Preparation of Methyl Esters of Compounds II
To a solution of 400 mg of lactone 2h from Example 2 in 100 ml of
absolute methanol is added 10 ml 0.1 M sodium methoxide in absolute
methanol. This solution is allowed to stand at room temperature for
one hour, then is diluted with water and extracted twice with ethyl
acetate. The organic phase is separated, dried (Na.sub.2 SO.sub.4),
filtered and evaporated in vacuo to yield the desired methyl
ester.
In like manner, by the use of equivalent amounts of propanol,
butanol, isobutanol, t-butanol, amylalcohol, isoamylalcohol,
2-dimethylaminoethanol, benzylalcohol, phenethanol,
2-acetamidoethanol and the like, and employing the corresponding
alcohol as solvent, the corresponding esters are obtained.
EXAMPLE 70
Preparation of Free Dihydroxy Acids
The sodium salt of the compound II from Example 64 is dissolved in
2 ml of ethanol-water (1:1; v:v) and added to 10 ml of 1N
hydrochloric acid from which the dihydroxy acid is extracted with
ethyl acetate. The organic extract is washed once with water, dried
(Na.sub.2 SO.sub.4), and evaporated in vacuo with a bath
temperature not exceeding 30.degree. C. The dihydroxy acid
derivative derived slowly reverts to the corresponding, parent
lactone on standing. The dihydroxy acid can be maintained by
increasing the pH above 7.0.
EXAMPLE 71
As a specific embodiment of a composition of this invention, 20 mg
of lactone 2h from Example 2, is formulated with sufficient finely
divided lactose to provide a total amount of 580 to 590 mg to fill
a size 0, hard gelatin capsule.
* * * * *