U.S. patent application number 11/355656 was filed with the patent office on 2006-06-22 for partial and full agonists of a1 adenosine receptors.
Invention is credited to Elfatih Elzein, Xiaofen Li, Venkata Palle, Jeff Zablocki.
Application Number | 20060135467 11/355656 |
Document ID | / |
Family ID | 31891396 |
Filed Date | 2006-06-22 |
United States Patent
Application |
20060135467 |
Kind Code |
A1 |
Zablocki; Jeff ; et
al. |
June 22, 2006 |
Partial and full agonists of A1 adenosine receptors
Abstract
Disclosed are novel compounds that are partial and full A.sub.1
adenosine receptor agonists, useful for treating various disease
states, in particular the supraventricular tachycardias, emesis,
angina, myocardial infarction and hyperlipidemia.
Inventors: |
Zablocki; Jeff; (Mountain
View, CA) ; Palle; Venkata; (Gurgaon, IN) ;
Elzein; Elfatih; (Fremont, CA) ; Li; Xiaofen;
(Palo Alto, CA) |
Correspondence
Address: |
Brian Lewis;CV Therapeutics, Inc.
3172 Porter Drive
Palo Alto
CA
94304
US
|
Family ID: |
31891396 |
Appl. No.: |
11/355656 |
Filed: |
February 15, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10641930 |
Aug 15, 2003 |
7022681 |
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11355656 |
Feb 15, 2006 |
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60403712 |
Aug 15, 2002 |
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60450094 |
Feb 25, 2003 |
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Current U.S.
Class: |
514/46 ;
536/27.3 |
Current CPC
Class: |
A61P 43/00 20180101;
A61P 9/06 20180101; C07H 19/16 20130101; A61P 3/06 20180101; A61P
25/08 20180101; A61P 9/00 20180101; A61P 37/06 20180101; A61P 9/10
20180101; A61P 3/04 20180101; A61P 3/10 20180101 |
Class at
Publication: |
514/046 ;
536/027.3 |
International
Class: |
A61K 31/7076 20060101
A61K031/7076; C07H 19/16 20060101 C07H019/16 |
Claims
1. A compound of the formula: ##STR32## wherein: R.sup.1 is
optionally substituted cycloalkyl, optionally substituted
heterocyclyl, optionally substituted aryl, or optionally
substituted heteroaryl; R.sup.2 is hydrogen, halo, trifluoromethyl,
or cyano; R.sup.3 is hydrogen, optionally substituted cycloalkyl,
optionally substituted aryl, optionally substituted heteroaryl, or
optionally substituted heterocyclyl, R.sup.4 and R.sup.5 are
independently hydrogen or optionally substituted acyl; X is a
covalent bond or lower alkylene optionally substituted by
cycloalkyl; X.sup.1 is a covalent bond or alkylene. Y is a covalent
bond or lower alkylene optionally substituted by hydroxy or
cycloalkyl; and Z is --C.ident.C--, --R.sup.6C.dbd.CR.sup.7--, or
--CHR.sup.6CHR.sup.7--, in which R.sup.6 and R.sup.7 at each
occurrence are hydrogen or lower alkyl.
2. The compound of claim 1, wherein Z is --C.ident.C--.
3. The compound of claim 2, wherein X, X.sup.1 and Y are covalent
bonds.
4. The compound of claim 3, wherein R.sup.1 is optionally
substituted cycloalkyl, optionally substituted heterocyclyl, or
optionally substituted heteroaryl.
5. The compound of claim 4, wherein R.sup.2, R.sup.4 and R.sup.5
are hydrogen.
6. The compound of claim 5, wherein R.sup.3 is hydrogen or
optionally substituted aryl.
7. The compound of claim 6, wherein R.sup.1 is optionally
substituted cyclopentyl or optionally substituted tetrahydrofuranyl
and R.sup.3 is hydrogen or optionally substituted phenyl.
8. The compound of claim 7, wherein R.sup.1 is cyclopentyl and
R.sup.3 is hydrogen, namely
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-ethynyloxolane-3,4-dio-
l.
9. The compound of claim 7, wherein R.sup.1 is 2-hydroxycyclopentyl
and R.sup.3 is hydrogen, namely
(4S,2R,3R,5R)-5-ethynyl-2-{6-[(2-hydroxycyclopentyl)amino]purin-9-yl}oxol-
ane-3,4-diol.
10. The compound of claim 7, wherein R.sup.1 is
tetrahydrofuran-3-yl and R.sup.3 is hydrogen, namely
(4S,2R,3R,5R)-2-[6-(oxolan-3-ylamino)purin-9-yl]-5-ethynyloxolane-3,4-dio-
l.
11. The compound of claim 7, wherein R.sup.1 is cyclopentyl and
R.sup.3 is 2-fluorophenyl, namely
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-[2-(2-fluorophenyl)eth-
ynyl]oxolane-3,4-diol.
12. The compound of claim 7, wherein R.sup.1 is cyclopentyl and
R.sup.3 is 2-trifluoromethylphenyl, namely
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{2-[2-(trifluoromethyl-
)-phenyl]ethynyl}oxolane-3,4-diol.
13. The compound of claim 7, wherein R.sup.1 is
tetrahydrofuran-3-yl and R.sup.3 is 2-fluorophenyl, namely
(4S,2R,3R,5R)-2-[6-(oxalan-3-ylamino)purin-9-yl]-5-{2-[2-fluorophenyl]eth-
ynyl}-oxolane-3,4-diol.
14. The compound of claim 7, wherein R.sup.1 is
tetrahydrofuran-3-yl and R.sup.3 is 2-trifluoromethylphenyl, namely
(4S,2R,3R,5R)-2-[6-(oxalan-3-ylamino)purin-9-yl]-5-{2-[2-(trifluoromethyl-
)phenyl]-ethynyl}oxolane-3,4-diol.
15. The compound of claim 5, wherein R.sup.3 is optionally
substituted heteroaryl.
16. The compound of claim 15, wherein R.sup.1 is optionally
substituted cyclopentyl or optionally substituted tetrahydrofuranyl
and R.sup.3 is optionally substituted thienyl.
17. The compound of claim 16, wherein R.sup.1 is cyclopentyl or
tetrahydrofuran-3-yl and R.sup.3 is thien-2-yl.
18. The compound of claim 1, wherein Z is
--R.sup.6C.dbd.CR.sup.7--, in which R.sup.6 and R.sup.7 are both
hydrogen.
19. The compound of claim 18, wherein X, X.sup.1 and Y are covalent
bonds.
20. The compound of claim 19, wherein R.sup.1 is optionally
substituted cycloalkyl.
21. The compound of claim 20, wherein R.sup.2, R.sup.4 and R.sup.5
are hydrogen.
22. The compound of claim 21, wherein R.sup.3 is optionally
substituted aryl or optionally substituted heteroaryl.
23. The compound of claim 22, wherein R.sup.1 is cyclopentyl and
R.sup.3 is 5-chlorothien-2-yl, 4-methylisoxazol-3-yl or
3,5-dimethylisoxazol-4-yl.
24. The compound of claim 22, wherein R.sup.1 is cyclopentyl and
R.sup.3 is 2-methyphenyl, namely
5-[2-(2-methylphenyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-y-
l]oxolane-3,4-diol.
25. The compound of claim 22, wherein R.sup.1 is cyclopentyl and
R.sup.3 is phenyl, namely
5-[2-(phenyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl
]oxolane-3,4-diol.
26. A method of treating a disease state in a mammal by
administration of an A.sub.1 adenosine receptor agonist, comprising
administering to a mammal in need thereof a therapeutically
effective dose of a compound of claim 1.
27. The method of claim 26, wherein the disease state is chosen
from atrial fibrillation, supraventricular tachycardia and atrial
flutter, congestive heart failure, epilepsy, stroke, diabetes,
obesity, ischemia, stable angina, unstable angina, cardiac
transplant, and myocardial infarction.
28. The method of claim 27, wherein the disease state is chosen
from atrial fibrillation, supraventricular tachycardia and atrial
flutter.
29. The method of claim 26, wherein the A.sub.1 adenosine receptor
agonist of claim 1 has an antilipolytic effect.
30. The method of claim 29, wherein said antilipolytic effect
treats disease states related to metabolic disorders
31. The method of claim 30, wherein the metabolic disorder is
hyperlipidemia, non-insulin-dependent diabetes mellitus, or
obesity.
32. A pharmaceutical composition comprising at least one
pharmaceutically acceptable excipient and a therapeutically
effective amount of a compound of Formula I. ##STR33## wherein:
R.sup.1 is optionally substituted cycloalkyl, optionally
substituted heterocyclyl, optionally substituted aryl, or
optionally substituted heteroaryl; R.sup.2 is hydrogen, halo,
trifluoromethyl, or cyano; R.sup.3 is optionally substituted
cycloalkyl, optionally substituted aryl, optionally substituted
heteroaryl, or optionally substituted heterocyclyl, R.sup.4 and
R.sup.5 are independently hydrogen or optionally substituted acyl;
X is a covalent bond or lower alkylene optionally substituted by
cycloalkyl; X.sup.1 is a covalent bond or alkylene; Y is a covalent
bond or lower alkylene optionally substituted by hydroxy or
cycloalkyl; and Z is --C.ident.C--, --R.sup.6C.dbd.CR.sup.7--, or
--CHR.sup.6CHR.sup.7--, in which R.sup.6 and R.sup.7 at each
occurrence are hydrogen or lower alkyl.
34. The method of claim 33, wherein the disease state is selected
from atrial fibrillation, supraventricular tachycardia and atrial
flutter.
35. The method of claim 33, wherein the disease state is chosen
from diabetes and obesity.
Description
[0001] Priority is claimed to U.S. Provisional Patent Application
Ser. No. 60/403,712, filed Aug. 15, 2002, and U.S. Provisional
Patent Application Ser. No. 60/450,094, filed Feb. 25, 2003, the
complete disclosures of which are hereby incorporated by
reference.
FIELD OF THE INVENTION
[0002] The present invention relates to novel compounds that are
partial or full A.sub.1 adenosine receptor agonists, and to their
use in treating mammals for various disease states, including
cardiovascular diseases, in particular arrhythmia and the
prevention of sudden death resulting from arrhythmia, ischemia, and
CNS disorders including pain, epilepsy, and emesis. A.sub.1
adenosine receptor agonists are antilipolytic agents, and are
useful for treating metabolic disorders, including diabetes and
obesity. The invention also relates to methods for their
preparation, and to pharmaceutical compositions containing such
compounds.
BACKGROUND
[0003] Adenosine is a naturally occurring nucleoside, which exerts
its biological effects by interacting with a family of adenosine
receptors known as A.sub.1, A.sub.2a, A.sub.2b, and A.sub.3, all of
which modulate important physiological processes. For example,
A.sub.2A adenosine receptors modulate coronary vasodilation,
A.sub.2B receptors have been implicated in mast cell activation,
asthma, vasodilation, regulation of cell growth, intestinal
function, and modulation of neurosecretion (See Adenosine A.sub.2B
Receptors as Therapeutic Targets, Drug Dev Res 45:198; Feoktistov
et al., Trends Pharmacol Sci 19:148-153), and A.sub.3 adenosine
receptors modulate cell proliferation processes.
[0004] A.sub.1 adenosine receptor agonists modulates the
cardiostimulatory effects of catecholamine (mediated via the
inhibition of adenylate cyclase), and slow the heart rate (HR) and
prolong impulse propagation through the AV node, which is due in
great part to activation of I.sub.KAdo. (B. Lerman and L.
Belardinelli Circulation, Vol. 83 (1991), P 1499-1509 and J. C.
Shryock and L. Belardinelli The Am. J. Cardiology, Vol. 79 (1997) P
2-10). Stimulation of the A.sub.1 adenosine receptor shortens the
duration and decreases the amplitude of the action potential of AV
nodal cells, and hence prolongs the refractory period of the AV
nodal cell. Thus, stimulation of A.sub.1 receptors provides a
method of treating supraventricular tachycardias, including
termination of nodal re-entrant tachycardias, and control of
ventricular rate during atrial fibrillation and flutter.
[0005] Elevated serum levels of non-esterified free fatty acid
(NEFA) are detrimental to both the mechanical and electrical
function of the heart, and A.sub.1 adenosine receptor agonists are
potent and efficacious inhibitors of lipolysis. Importantly,
because A.sub.1 adenosine receptor agonists are more potent in
adipose tissue that in heart tissues, they decrease lipolysis at
concentrations that do not affect heart rate. Thus, A.sub.1
adenosine receptor agonists are useful for treating metabolic
disorders such as non-insulin-dependent diabetes mellitus and
obesity via their anti-lipolytic activity. The antilipolytic effect
of adenosine A.sub.1 receptor agonists is also useful in the
management of congestive heart failure. Furthermore, A.sub.1
adenosine receptor agonists are protective against cardioischemia.
A.sub.1 adenosine receptor agonists are also useful as
chemotherapeutics in the treatment of CNS disorders including
epilepsy (anticonvulsant activity) and ischemia.
[0006] Accordingly, it is an object of this invention to provide
compounds that are potent full A.sub.1 adenosine receptor agonists
or partial A.sub.1 adenosine receptor agonists. Preferred compounds
of the invention are selective for the A.sub.1 adenosine receptor,
which minimizes undesired side effects related to stimulation or
antagonism of the other adenosine receptors.
SUMMARY OF THE INVENTION
[0007] Accordingly, in a first aspect, the invention relates to
compounds of Formula I: ##STR1## [0008] wherein: [0009] R.sup.1 is
optionally substituted cycloalkyl, optionally substituted
heterocyclyl, optionally substituted aryl, or optionally
substituted heteroaryl; [0010] R.sup.2 is hydrogen, halo,
trifluoromethyl, or cyano; [0011] R.sup.3 is hydrogen, optionally
substituted cycloalkyl, optionally substituted aryl, optionally
substituted heteroaryl, or optionally substituted heterocyclyl,
[0012] R.sup.4 and R.sup.5 are independently hydrogen or optionally
substituted acyl; [0013] X is a covalent bond or lower alkylene
optionally substituted by cycloalkyl; [0014] X.sup.1 is a covalent
bond or alkylene. [0015] Y is a covalent bond or lower alkylene
optionally substituted by hydroxy or cycloalkyl; and [0016] Z is
--C.ident.C--, --R.sup.6C.dbd.CR.sup.7--, or
--CHR.sup.6CHR.sup.7--, in which R.sup.6 and R.sup.7 at each
occurrence are hydrogen or lower alkyl
[0017] A second aspect of the invention relates to pharmaceutical
formulations, comprising a therapeutically effective amount of a
compound of Formula I and at least one pharmaceutically acceptable
excipient.
[0018] A third aspect of the invention relates to a method of using
the compounds of Formula I in the treatment of a disease or
condition in a mammal that can be effectively treated with a
partial or full selective A.sub.1 adenosine receptor agonist. Such
diseases and conditions include at least one of the following;
supraventricular tachycardia, including atrial fibrillation, and
atrial flutter, ischemia, including that due to stable and unstable
angina, congestive heart failure, myocardial infarction, disorders
of the CNS including epilepsy and stroke, metabolic disorders, such
as obesity and diabetes, or the sequela of diabetes or congestive
heart failure specifically hyperlipidemia, which is alleviated by
the antilipolytic effect of A.sub.1 agonists on adipocytes; and the
treatment of nausea (emesis).
[0019] A fourth aspect of this invention relates to methods of
preparing the compounds of Formula I.
[0020] Of the compounds of Formula I, one preferred class includes
those compounds in which Z is --C.ident.C--, particularly those
compounds in which X, X.sup.1 and Y are covalent bonds. A preferred
group within this class includes those compounds in which R.sup.1
is optionally substituted cycloalkyl optionally substituted
heterocyclyl, or optionally substituted heteroaryl, and R.sup.2,
R.sup.4 and R.sup.5 are hydrogen.
[0021] A preferred subgroup includes those compounds of Formula I
in which R.sup.3 is hydrogen or optionally substituted aryl,
especially optionally substituted phenyl. Particularly preferred
compounds within this subgroup are those compounds in which R.sup.1
is cycloalkyl, especially cyclopentyl or hydroxycyclopentyl, or
optionally substituted heterocyclyl, especially
tetrahydrofuran-3-yl, and R.sup.3 is hydrogen. Other preferred
compounds within this subgroup includes those compounds of Formula
I in which R.sup.3 is optionally substituted phenyl. Particularly
preferred are those compounds in which R.sup.1 is cycloalkyl,
especially cyclopentyl, or optionally substituted heterocyclyl,
especially tetrahydrofuran-3-yl, and R.sup.3is 2-fluorophenyl or
2-trifluoromethylphenyl.
[0022] Another preferred subgroup includes those compounds of
Formula I in which R.sup.3 is optionally substituted aryl.
Particularly preferred compounds within this subgroup are those
compounds in which R.sup.1 is cycloalkyl, especially cyclopentyl,
or optionally substituted heterocyclyl, especially
tetrahydrofuran-3-yl. Preferred R.sup.3 groups include optionally
substituted thienyl, especially 5-chlorothien-2-yl.
[0023] Of the compounds of Formula I, another preferred class
includes those compounds in which Z is --R.sup.6C.dbd.CR.sup.7--,
particularly those compounds in which R.sup.6 and R.sup.7 are
hydrogen and X, X and Y are covalent bonds. A preferred group
within this class includes those compounds in which R.sup.1 is
optionally substituted cycloalkyl, and R.sup.2, R.sup.4 and R.sup.5
are hydrogen. A preferred subgroup includes those compounds of
Formula I in which R.sup.3 is optionally substituted aryl or
optionally substituted heteroaryl. Particularly preferred are those
compounds in which R.sup.3 includes optionally substituted phenyl,
especially phenyl or 2-methylphenyl, or optionally substituted
thienyl, especially 5-chlorothien-2-yl.
Definitions and General Parameters
[0024] As used in the present specification, the following words
and phrases are generally intended to have the meanings as set
forth below, except to the extent that the context in which they
are used indicates otherwise.
[0025] The term "alkyl" refers to a monoradical branched or
unbranched saturated hydrocarbon chain having from 1 to 20 carbon
atoms. This term is exemplified by groups such as methyl, ethyl,
n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-hexyl,
n-decyl, tetradecyl, and the like.
[0026] The term "substituted alkyl" refers to: [0027] 1) an alkyl
group as defined above, having 1, 2, 3, 4 or 5 substituents, for
example 1 to 3 substituents, selected from the group consisting of
alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl,
acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino,
azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy,
carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol,
alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl,
aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy,
hydroxyamino, alkoxyamino, nitro, --SO-alkyl, --SO-aryl,
--SO-heteroaryl, --SO.sub.2-alkyl, SO.sub.2-aryl and
--SO.sub.2-heteroaryl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1, 2, or 3 substituents chosen from alkyl, carboxy,
carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3,
amino, substituted amino, cyano, and --S(O).sub.nR, where R is
alkyl, aryl, or heteroaryl and n is 0, 1 or 2; or [0028] 2) an
alkyl group as defined above that is interrupted by 1-10 atoms
independently chosen from oxygen, sulfur and NR.sub.a--, where
R.sub.a is chosen from hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclyl. All
substituents may be optionally further substituted by alkyl,
alkoxy, halogen, CF.sub.3, amino, substituted amino, cyano, or
--S(O).sub.nR, in which R is alkyl, aryl, or heteroaryl and n is 0,
1 or 2; or [0029] 3) an alkyl group as defined above that has both
1, 2, 3, 4 or 5 substituents as defined above and is also
interrupted by 1-10 atoms as defined above.
[0030] The term "lower alkyl" refers to a monoradical branched or
unbranched saturated hydrocarbon chain having 1, 2, 3, 4, 5, or 6
carbon atoms. This term is exemplified by groups such as methyl,
ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, t-butyl, n-hexyl,
and the like.
[0031] The term "substituted lower alkyl" refers to lower alkyl as
defined above having 1 to 5 substituents, for example 1, 2, or 3
substituents, as defined for substituted alkyl, or a lower alkyl
group as defined above that is interrupted by 1, 2, 3, 4, or 5
atoms as defined for substituted alkyl, or a lower alkyl group as
defined above that has both 1, 2, 3, 4 or 5 substituents as defined
above and is also interrupted by 1, 2, 3, 4, or 5 atoms as defined
above.
[0032] The term "alkylene" refers to a diradical of a branched or
unbranched saturated hydrocarbon chain, for example having from 1
to 20 carbon atoms, preferably 1-10 carbon atoms, more preferably
1, 2, 3, 4, 5 or 6 carbon atoms. This term is exemplified by groups
such as methylene (--CH.sub.2--), ethylene (--CH.sub.2CH.sub.2--),
the propylene isomers (e.g., --CH.sub.2CH.sub.2CH.sub.2-- and
--CH(CH.sub.3)CH.sub.2--) and the like.
[0033] The term "lower alkylene" refers to a diradical of a
branched or unbranched saturated hydrocarbon chain, for example
having from 1, 2, 3, 4, 5, or 6 carbon atoms.
[0034] The term "substituted alkylene" refers to: [0035] (1) an
alkylene group as defined above having 1, 2, 3, 4, or 5
substituents selected from the group consisting of alkyl, alkenyl,
alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino,
acyloxy, amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano,
halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl,
arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl,
aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino,
heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino,
alkoxyamino, nitro, --SO-alkyl, --SO-aryl, --SO-heteroaryl,
--SO.sub.2-alkyl, SO.sub.2-aryl and --SO.sub.2-heteroaryl. Unless
otherwise constrained by the definition, all substituents may
optionally be further substituted by 1, 2, or 3 substituents chosen
from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy,
halogen, CF.sub.3, amino, substituted amino, cyano, and
--S(O).sub.nR, where R is alkyl, aryl, or heteroaryl and n is 0, 1
or 2; or [0036] (2) an alkylene group as defined above that is
interrupted by 1-20 atoms independently chosen from oxygen, sulfur
and NR.sub.a--, where R.sub.a is chosen from hydrogen, optionally
substituted alkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl and
heterocycyl, or groups selected from carbonyl, carboxyester,
carboxyamide and sulfonyl; or [0037] (3) an alkylene group as
defined above that has both 1, 2, 3, 4 or 5 substituents as defined
above and is also interrupted by 1-20 atoms as defined above.
Examples of substituted alkylenes are chloromethylene (--CH(Cl)--),
aminoethylene (--CH(NH.sub.2)CH.sub.2--), methylaminoethylene
(--CH(NHMe)CH.sub.2--), 2-carboxypropylene isomers
(--CH.sub.2CH(CO.sub.2H)CH.sub.2--), ethoxyethyl
(--CH.sub.2CH.sub.2O--CH.sub.2CH.sub.2--), ethylmethylaminoethyl
(--CH.sub.2CH.sub.2N(CH.sub.3)CH.sub.2CH.sub.2--),
1-ethoxy-2-(2-ethoxy-ethoxy)ethane
(--CH.sub.2CH.sub.2O--CH.sub.2CH.sub.2--OCH.sub.2CH.sub.2--OCH.sub.2CH.su-
b.2--), and the like.
[0038] The term "aralkyl" refers to an aryl group covalently linked
to an alkylene group, where aryl and alkylene are defined herein.
"Optionally substituted aralkyl" refers to an optionally
substituted aryl group covalently linked to an optionally
substituted alkylene group. Such aralkyl groups are exemplified by
benzyl, phenylethyl, 3-(4-methoxyphenyl)propyl, and the like.
[0039] The term "alkoxy" refers to the group R--O--, where R is
optionally substituted alkyl or optionally substituted cycloalkyl,
or R is a group --Y-Z, in which Y is optionally substituted
alkylene and Z is optionally substituted alkenyl, optionally
substituted alkynyl; or optionally substituted cycloalkenyl, where
alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl are as defined
herein. Preferred alkoxy groups are alkyl-O-- and include, by way
of example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy,
tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy,
and the like.
[0040] The term "alkylthio" refers to the group R--S--, where R is
as defined for alkoxy.
[0041] The term "alkenyl" refers to a monoradical of a branched or
unbranched unsaturated hydrocarbon group having from 2 to 20 carbon
atoms, preferably 2 to 10 carbon atoms,and even more preferably 2
to 6 carbon atoms and having 1-6, preferably 1, double bond
(vinyl). Preferred alkenyl groups include ethenyl or vinyl
(--CH.dbd.CH.sub.2), 1-propylene or allyl
(--CH.sub.2CH.dbd.CH.sub.2), isopropylene
(--C(CH.sub.3).dbd.CH.sub.2), bicyclo[2.2.1]heptene, and the like.
In the event that alkenyl is attached to nitrogen, the double bond
cannot be alpha to the nitrogen.
[0042] The term "lower alkenyl" refers to alkenyl as defined above
having from 2 to 6 carbon atoms.
[0043] The term "substituted alkenyl" refers to an alkenyl group as
defined above having 1, 2, 3, 4 or 5 substituents, and preferably
1, 2, or 3 substituents, selected from the group consisting of
alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl,
acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino,
azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy,
carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol,
alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl,
aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy,
hydroxyamino, alkoxyamino, nitro, --SO-alkyl, --SO-aryl,
--SO-heteroaryl, --SO.sub.2-alkyl, SO.sub.2-aryl and
--SO.sub.2-heteroaryl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1, 2, or 3 substituents chosen from alkyl, carboxy,
carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3,
amino, substituted amino, cyano, and --S(O).sub.nR, where R is
alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
[0044] The term "alkynyl" refers to a monoradical of an unsaturated
hydrocarbon having from 2 to 20 carbon atoms, preferably 2 to 10
carbon atoms and even more preferably 2 to 6 carbon atoms and
having at least 1 and preferably from 1-6 sites of acetylene
(triple bond) unsaturation. Preferred alkynyl groups include
ethynyl, (--C.ident.CH), propargyl (or propynyl,
--C.ident.CCH.sub.3), and the like. In the event that alkynyl is
attached to nitrogen, the triple bond cannot be alpha to the
nitrogen.
[0045] The term "substituted alkynyl" refers to an alkynyl group as
defined above having 1, 2, 3, 4 or 5 substituents, and preferably
1, 2, or 3 substituents, selected from the group consisting of
alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl,
acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino,
azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy,
carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol,
alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl,
aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy,
hydroxyamino, alkoxyamino, nitro, --SO-alkyl, --SO-aryl,
--SO-heteroaryl, --SO.sub.2-alkyl, SO.sub.2-aryl and
--SO.sub.2-heteroaryl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1, 2, or 3 substituents chosen from alkyl, carboxy,
carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3,
amino, substituted amino, cyano, and --S(O).sub.nR, where R is
alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
[0046] The term "aminocarbonyl" refers to the group --C(O)NRR where
each R is independently hydrogen, alkyl, cycloaklyl, aryl,
heteroaryl, heterocyclyl or where both R groups are joined to form
a heterocyclic group (e.g., morpholino). Unless otherwise
constrained by the definition, all substituents may optionally be
further substituted by 1, 2, or 3 substituents chosen from alkyl,
carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen,
CF.sub.3, amino, substituted amino, cyano, and --S(O).sub.nR, where
R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
[0047] The term "ester" or "carboxyester" refers to the group
--C(O)OR, where R is alkyl, cycloalkyl, aryl, heteroaryl, or
heterocyclyl, which may be optionally further substituted by alkyl,
alkoxy, halogen, CF.sub.3, amino, substituted amino, cyano, or
--S(O).sub.nR.sub.a, in which R.sub.a is alkyl, aryl, or heteroaryl
and n is 0, 1 or 2.
[0048] The term "acylamino" refers to the group --NRC(O)R where
each R is independently hydrogen, alkyl, aryl, heteroaryl, or
heterocyclyl. All substituents may be optionally further
substituted by alkyl, alkoxy, halogen, CF.sub.3, amino, substituted
amino, cyano, or --S(O).sub.nR, in which R is alkyl, aryl, or
heteroaryl and n is 0, 1 or 2.
[0049] The term "acyloxy" refers to the groups --O(O)C-alkyl,
--O(O)C-cycloalkyl, --O(O)C-aryl, --O(O)C-heteroaryl, and
--O(O)C-heterocyclyl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1, 2, or 3 substituents chosen from alkyl, carboxy,
carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3,
amino, substituted amino, cyano, and --S(O).sub.nR, where R is
alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
[0050] The term "aryl" refers to an aromatic carbocyclic group of 6
to 20 carbon atoms having a single ring (e.g., phenyl) or multiple
rings (e.g., biphenyl), or multiple condensed (fused) rings (e.g.,
naphthyl or anthryl). Preferred aryls include phenyl, naphthyl and
the like.
[0051] Unless otherwise constrained by the definition for the aryl
substituent, such aryl groups can optionally be substituted with 1,
2, 3, 4 or 5 substituents, preferably 1, 2, or 3 substituents,
selected from the group consisting of alkyl, alkenyl, alkynyl,
alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, amino,
aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy,
keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio,
heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy,
heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy,
heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro,
--SO-alkyl, --SO-aryl, --SO-heteroaryl, --SO.sub.2-alkyl,
SO.sub.2-aryl and --SO.sub.2-heteroaryl. Unless otherwise
constrained by the definition, all substituents may optionally be
further substituted by 1, 2, or 3 substituents chosen from alkyl,
carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen,
CF.sub.3, amino, substituted amino, cyano, and --S(O).sub.nR, where
R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
[0052] The term "aryloxy" refers to the group aryl-O-- wherein the
aryl group is as defined above, and includes optionally substituted
aryl groups as also defined above. The term "arylthio" refers to
the group R--S--, where R is as defined for aryl.
[0053] The term "amino" refers to the group --NH.sub.2.
[0054] The term "substituted amino" refers to the group --NRR where
each R is independently selected from the group consisting of
hydrogen, alkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl
provided that both R groups are not hydrogen, or a group --Y-Z, in
which Y is optionally substituted alkylene and Z is alkenyl,
cycloalkenyl, or alkynyl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1, 2, or 3 substituents chosen from alkyl, carboxy,
carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3,
amino, substituted amino, cyano, and --S(O).sub.nR, where R is
alkyl, aryl, or heteroaryl and n is 0, 1 or2.
[0055] The term "carboxyalkyl" refers to the groups --C(O)O-alkyl,
--C(O)O-cycloalkyl, where alkyl and cycloalkyl, are as defined
herein, and may be optionally further substituted by alkyl,
alkenyl, alkynyl, alkoxy, halogen, CF.sub.3, amino, substituted
amino, cyano, or --S(O).sub.nR, in which R is alkyl, aryl, or
heteroaryl and n is 0, 1 or 2.
[0056] The term "cycloalkyl" refers to cyclic alkyl groups of from
3 to 20 carbon atoms having a single cyclic ring or multiple
condensed rings. Such cycloalkyl groups include, by way of example,
single ring structures such as cyclopropyl, cyclobutyl,
cyclopentyl, cyclooctyl, and the like, or multiple ring structures
such as adanantanyl, and bicyclo[2.2.1]heptane, or cyclic alkyl
groups to which is fused an aryl group, for example indan, and the
like.
[0057] The term "substituted cycloalkyl" refers to cycloalkyl
groups having 1, 2, 3, 4 or 5 substituents, and preferably 1, 2, or
3 substituents, selected from the group consisting of alkyl,
alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl,
acylamino, acyloxy, amino, aminocarbonyl, alkoxycarbonylamino,
azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy,
carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol,
alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl,
aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy,
hydroxyamino, alkoxyamino, nitro, --SO-alkyl, --SO-aryl,
--SO-heteroaryl, --SO.sub.2-alkyl, SO.sub.2-aryl and
--SO.sub.2-heteroaryl. Unless otherwise constrained by the
definition, all substituents may optionally be further substituted
by 1, 2, or 3 substituents chosen from alkyl, carboxy,
carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF.sub.3,
amino, substituted amino, cyano, and --S(O).sub.nR, where R is
alkyl, aryl, or heteroaryl and n is 0, 1 or 2.
[0058] The term "halogen" or "halo" refers to fluoro, bromo,
chloro, and iodo.
[0059] The term "acyl" denotes a group --C(O)R, in which R is
hydrogen, optionally substituted alkyl, optionally substituted
cycloalkyl, optionally substituted heterocyclyl, optionally
substituted aryl, and optionally substituted heteroaryl.
[0060] The term "heteroaryl" refers to an aromatic group (i.e.,
unsaturated) comprising 1 to 15 carbon atoms and 1 to 4 heteroatoms
selected from oxygen, nitrogen and sulfur within at least one
ring.
[0061] Unless otherwise constrained by the definition for the
heteroaryl substituent, such heteroaryl groups can be optionally
substituted with 1 to 5 substituents, preferably 1, 2, or 3
substituents selected from the group consisting of alkyl, alkenyl,
alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino,
acyloxy, amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano,
halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl (an
alkyl ester), arylthio, heteroaryl, heteroarylthio,
heterocyclylthio, thiol, alkylthio, aryl, aryloxy, aralkyl,
heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy,
heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro,
--SO-alkyl, --SO-aryl, --SO-heteroaryl, --SO.sub.2-alkyl,
SO.sub.2-aryl and --SO.sub.2-heteroaryl. Unless otherwise
constrained by the definition, all substituents may optionally be
further substituted by 1, 2, or 3 substituents chosen from alkyl,
carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen,
CF.sub.3, amino, substituted amino, cyano, and --S(O).sub.nR, where
R is alkyl, aryl, or heteroaryl and n is 0, 1 or 2. Such heteroaryl
groups can have a single ring (e.g., pyridyl or furyl) or multiple
condensed rings (e.g., indolizinyl, benzothiazole, or
benzothienyl). Examples of nitrogen heterocycles and heteroaryls
include, but are not limited to, pyrrole, imidazole, pyrazole,
pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole,
indole, indazole, purine, quinolizine, isoquinoline, quinoline,
phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline,
pteridine, carbazole, carboline, phenanthridine, acridine,
phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine,
phenothiazine, imidazolidine, imidazoline, and the like as well as
N-alkoxy-nitrogen containing heteroaryl compounds.
[0062] The term "heteroaryloxy" refers to the group
heteroaryl-O--.
[0063] The term "heterocyclyl" refers to a monoradical saturated or
partially unsaturated group having a single ring or multiple
condensed rings, having from 1 to 40 carbon atoms and from 1 to 10
hetero atoms, preferably 1 to 4 heteroatoms, selected from
nitrogen, sulfur, phosphorus, and/or oxygen within the ring.
[0064] Unless otherwise constrained by the definition for the
heterocyclic substituent, such heterocyclic groups can be
optionally substituted with 1 to 5, and preferably 1, 2, or 3
substituents, selected from the group consisting of alkyl, alkenyl,
alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino,
acyloxy, amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano,
halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl,
arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl,
aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino,
heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino,
alkoxyamino, nitro, --SO-alkyl, --SO-aryl, --SO-heteroaryl,
--SO.sub.2-alkyl, SO.sub.2-aryl and --SO.sub.2-heteroaryl. Unless
otherwise constrained by the definition, all substituents may
optionally be further substituted by 1, 2, or 3 substituents chosen
from alkyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy,
halogen, CF.sub.3, amino, substituted amino, cyano, and
--S(O).sub.nR, where R is alkyl, aryl, or heteroaryl and n is 0, 1
or 2. Heterocyclic groups can have a single ring or multiple
condensed rings. Preferred heterocyclics include tetrahydrofuranyl,
morpholino, piperidinyl, and the like.
[0065] The term "thiol" refers to the group --SH.
[0066] The term "substituted alkylthio" refers to the group
--S-substituted alkyl.
[0067] The term "heteroarylthiol" refers to the group
--S-heteroaryl wherein the heteroaryl group is as defined above
including optionally substituted heteroaryl groups as also defined
above.
[0068] The term "sulfoxide" refers to a group --S(O)R, in which R
is alkyl, aryl, or heteroaryl. "Substituted sulfoxide" refers to a
group --S(O)R, in which R is substituted alkyl, substituted aryl,
or substituted heteroaryl, as defined herein.
[0069] The term "sulfone" refers to a group --S(O).sub.2R, in which
R is alkyl, aryl, or heteroaryl. "Substituted sulfone" refers to a
group --S(O).sub.2R, in which R is substituted alkyl, substituted
aryl, or substituted heteroaryl, as defined herein.
[0070] The term "keto" refers to a group --C(O)--. The term
"thiocarbonyl" refers to a group --C(S)--. The term "carboxy"
refers to a group --C(O)--OH.
[0071] "Optional" or "optionally" means that the subsequently
described event or circumstance may or may not occur, and that the
description includes instances where said event or circumstance
occurs and instances in which it does not.
[0072] The term "compound of Formula I" is intended to encompass
the compounds of the invention as disclosed and polymorphs thereof,
pharmaceutcally acceptable salts, pharmaceutically acceptable
esters, and prodrugs of such compounds. Additionally, the compounds
of the invention may possess one or more asymmetric centers, and
can be produced as a racemic mixture or as individual enantiomers
or diastereoisomers. The number of stereoisomers present in any
given compound of Formula I depends upon the number of asymmetric
centers present (there are 2.sup.n stereoisomers possible where n
is the number of asymmetric centers). The individual stereoisomers
may be obtained by resolving a racemic or non-racemic mixture of an
intermediate at some appropriate stage of the synthesis, or by
resolution of the compound of Formula I by conventional means. The
individual stereoisomers (including individual enantiomers and
diastereoisomers) as well as racemic and non-racemic mixtures of
stereoisomers are encompassed within the scope of the present
invention, all of which are intended to be depicted by the
structures of this specification unless otherwise specifically
indicated.
[0073] "Isomers" are different compounds that have the same
molecular formula.
[0074] "Stereoisomers" are isomers that differ only in the way the
atoms are arranged in space.
[0075] "Enantiomers" are a pair of stereoisomers that are
non-superimposable mirror images of each other. A 1:1 mixture of a
pair of enantiomers is a "racemic" mixture. The term "(.+-.)" is
used to designate a racemic mixture where appropriate.
[0076] "Diastereoisomers" are stereoisomers that have at least two
asymmetric atoms, but which are not mirror-images of each
other.
[0077] The absolute stereochemistry is specified according to the
Cahn-Ingold-Prelog R-S system. When the compound is a pure
enantiomer the stereochemistry at each chiral carbon may be
specified by either R or S. Resolved compounds whose absolute
configuration is unknown are designated (+) or (-) depending on the
direction (dextro- or laevorotary) which they rotate the plane of
polarized light at the wavelength of the sodium D line.
[0078] The term "therapeutically effective amount" refers to that
amount of a compound of Formula I that is sufficient to effect
treatment, as defined below, when administered to a mammal in need
of such treatment. The therapeutically effective amount will vary
depending upon the subject and disease condition being treated, the
weight and age of the subject, the severity of the disease
condition, the manner of administration and the like, which can
readily be determined by one of ordinary skill in the art.
[0079] The term "treatment" or "treating" means any treatment of a
disease in a mammal, including: [0080] (i) preventing the disease,
that is, causing the clinical symptoms of the disease not to
develop; [0081] (ii) inhibiting the disease, that is, arresting the
development of clinical symptoms; and/or [0082] (iii) relieving the
disease, that is, causing the regression of clinical symptoms.
[0083] In many cases, the compounds of this invention are capable
of forming acid and/or base salts by virtue of the presence of
amino and/or carboxyl groups or groups similar thereto. The term
"pharmaceutically acceptable salt" refers to salts that retain the
biological effectiveness and properties of the compounds of Formula
I, and which are not biologically or otherwise undesirable.
Pharmaceutically acceptable base addition salts can be prepared
from inorganic and organic bases. Salts derived from inorganic
bases, include by way of example only, sodium, potassium, lithium,
ammonium, calcium and magnesium salts. Salts derived from organic
bases include, but are not limited to, salts of primary, secondary
and tertiary amines, such as alkyl amines, dialkyl amines, trialkyl
amines, substituted alkyl amines, di(substituted alkyl)amines,
tri(substituted alkyl)amines, alkenyl amines, dialkenyl amines,
trialkenyl amines, substituted alkenyl amines, di(substituted
alkenyl)amines, tri(substituted alkenyl)amines, cycloalkyl amines,
di(cycloalkyl)amines, tri(cycloalkyl)amines, substituted cycloalkyl
amines, disubstituted cycloalkyl amine, trisubstituted cycloalkyl
amines, cycloalkenyl amines, di(cycloalkenyl)amines,
tri(cycloalkenyl)amines, substituted cycloalkenyl amines,
disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl
amines, aryl amines, diaryl amines, triaryl amines, heteroaryl
amines, diheteroaryl amines, triheteroaryl amines, heterocyclic
amines, diheterocyclic amines, triheterocyclic amines, mixed di-
and tri-amines where at least two of the substituents on the amine
are different and are selected from the group consisting of alkyl,
substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl,
substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl,
aryl, heteroaryl, heterocyclic, and the like. Also included are
amines where the two or three substituents, together with the amino
nitrogen, form a heterocyclic or heteroaryl group.
[0084] Specific examples of suitable amines include, by way of
example only, isopropylamine, trimethyl amine, diethyl amine,
tri(iso-propyl)amine, tri(n-propyl)amine, ethanolamine,
2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine,
caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine,
glucosamine, N-alkylglucamines, theobromine, purines, piperazine,
piperidine, morpholine, N-ethylpiperidine, and the like.
[0085] Pharmaceutically acceptable acid addition salts may be
prepared from inorganic and organic acids. Salts derived from
inorganic acids include hydrochloric acid, hydrobromic acid,
sulfuric acid, nitric acid, phosphoric acid, and the like. Salts
derived from organic acids include acetic acid, propionic acid,
glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid,
succinic acid, maleic acid, fumaric acid, tartaric acid, citric
acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic
acid, ethanesulfonic acid, p-toluene-sulfonic acid, salicylic acid,
and the like.
[0086] As used herein, "pharmaceutically acceptable carrier"
includes any and all solvents, dispersion media, coatings,
antibacterial and antifungal agents, isotonic and absorption
delaying agents and the like. The use of such media and agents for
pharmaceutically active substances is well known in the art. Except
insofar as any conventional media or agent is incompatible with the
active ingredient, its use in the therapeutic compositions is
contemplated. Supplementary active ingredients can also be
incorporated into the compositions.
[0087] As used herein, the term "agonist" refers to the ability of
a compound to interact with a receptor and evoke a maximal
physiological effect (that is, activate or stimulate the receptor).
This effect is known as the intrinsic efficacy. Many full agonists
of the adenosine A.sub.1 receptor are known to those skilled in the
art, for example N6-cyclopentyladenosine (CPA, or CCPA). Some
adenosine A.sub.1 agonists are referred to as "partial agonists"
because they interact with adenosine A.sub.1 receptors but produce
a less than maximal response when compared to an agonist such as
CPA.
[0088] The intrinsic efficacy of a compound is its differential
effect on a selected tissue. Thus, a compound may be a full agonist
in a given tissue but a partial in others. The compounds identified
by this invention have therapeutically useful affinities for the
adenosine A.sub.1 receptor but have a range of intrinsic efficacies
from full agonist to partial agonist. That is, some compounds may
have no effect with respect to a given effector system in a given
cell type, but be a full agonist in another cell type and/or
effector system. A partial agonist targeted to a selected target is
likely to cause fewer side effects than a full agonist, because
they will be less likely to induce desensitization of the A.sub.1
receptor (R. B. Clark, B. J. Knoll, R. Barber TiPS, Vol. 20 (1999)
p. 279-286) and to cause side effects. Chronic administration of a
full agonist (R-N6-phenylisopropyladenosine, R-PIA) for 7 days led
to a desensitization of the A.sub.1 receptor in terms of the
dromotropic response in guinea pigs (note: a decrease in receptor
number was observed--D. M. Dennis, J. C. Shryock, L. Belardinelli
JPET, Vol. 272 (1995) p. 1024-1035). The A.sub.1 agonist induced
inhibitory effect on the production of cAMP by adenylate cyclase in
adipocytes has been shown to desensitize upon chronic treatment
with an A.sub.1 agonist as well (W. J. Parsons and G. L. J. Biol.
Chem. Vol. 262 (1987) p. 841-847).
Nomenclature
[0089] The naming and numbering of the compounds of the invention
is illustrated with a representative compound of Formula I in which
R.sup.1 is cyclopentyl, R.sup.2 is hydrogen, R.sup.3 is
2-fluorophenyl, R.sup.4 and R.sup.5 are both hydrogen, X, X,.sup.1
and Y are covalent bonds, and Z is --C.ident.C--: ##STR2## which is
named:
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-[2-(2-fluorophenyl)eth-
ynyl]oxolane-3,4-diol, or alternatively may be named:
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)-9H-purin-9-yl]-5-[2-(2-fluorophenyl-
)ethynyl]tetrahydrofuran-3,4-diol. Synthetic Reaction
Parameters
[0090] The terms "solvent", "inert organic solvent" or "inert
solvent" mean a solvent inert under the conditions of the reaction
being described in conjunction therewith [including, for example,
benzene, toluene, acetonitrile, tetrahydrofuran ("THF"),
dimethylformamide ("DMF"), chloroform, methylene chloride (or
dichloromethane), diethyl ether, methanol, pyridine and the like].
Unless specified to the contrary, the solvents used in the
reactions of the present invention are inert organic solvents.
[0091] The term "q.s." means adding a quantity sufficient to
achieve a stated function, e.g., to bring a solution to the desired
volume (i.e., 100%).
Synthesis of the Compounds of Formula I
[0092] The compounds of Formula I where R.sup.3 is hydrogen,
X.sup.1 and Y are covalent bonds, and Z is --C.ident.C-- are
prepared starting from a compound of formula (1) as shown in
Reaction Scheme I. ##STR3## ##STR4## Step 1--Preparation of Formula
(2)
[0093] The starting compounds of formula (1) are commercially
available (for example, the compound of formula (1) in which
R.sup.2 is hydrogen is available from Aldrich, Milwaukee), or are
prepared by means well known to those in the art. The compound of
formula (2) is prepared conventionally from the compound of formula
(1) by reaction with 2,2-dimethoxypropane in an inert solvent,
preferably N,N-dimethylformamide, in the presence of a catalytic
amount of an acid catalyst, preferably p-toluenesulfonic acid, at a
temperature of about 40-90.degree. C., preferably about 70.degree.
C., for about 24-72 hours, preferably about 48 hours. When the
reaction is substantially complete, the product of formula (2) is
isolated by conventional means, for example removal of the solvent
under reduced pressure and purifying the residue by
chromatography.
Step 2--Preparation of Formula (3)
[0094] The 6-chloro moiety is displaced from the compound of
formula (2) by reaction with a compound of formula
R.sup.1XNH.sub.2, where X is as defined above, in the presence of a
base, for example triethylamine. The reaction is carried out in an
inert protic solvent, for example ethanol, at a temperature of
about reflux, for about 14-48 hours, preferably about 16 hours.
When the reaction is substantially complete, the product of formula
(3) is isolated by conventional means, for example by removal of
the solvent under reduced pressure, followed by crystallization of
the residue from a suitable solvent.
[0095] It should be noted that steps 1 and 2 may be carried out in
reverse order.
Step 3--Preparation of Formula (4)
[0096] The hydroxymethyl compound of formula (3) is oxidized to an
aldehyde of formula (4) using a modification of the Moffat
Oxidation. In general, to the compound of formula (3) is added a
mixture of 1,3-dicyclohexylcarbodimide DCC, dimethysulfoxide and
pyridine. The initial reaction is carried out at a temperature of
about -5.degree. to about 10.degree. C., preferably about 0.degree.
C., and then at about room temperature for about 6-48 hours,
preferably about 18 hours. When the reaction is substantially
complete, the aldehyde of formula (4) is isolated by conventional
means, for example by partitioning the product between ethyl
acetate and water and removing the solvent under reduced pressure.
The product is used in the next step without further
purification.
Step 4--Preparation of Formula (5).
[0097] The 4'-aldehyde group is converted to an ethynyl group by
reaction with bromomethyltriphenylphosphonium bromide in the
presence of a strong base, preferably potassium t-butoxide. The
reaction is carried out in an inert solvent, preferably
tetrahydrofuran, at a temperature of about -80.degree. C., allowing
the reaction mixture to gradually warm to room temperature over a
period of about 1-3 days. When the reaction is substantially
complete, the product of formula (5) is isolated by conventional
means, for example by removal of the solvent under reduced
pressure, followed by partition between a solvent such as ethyl
acetate and water, removing the solvent under reduced pressure. The
residue may then be further purified by chromatography on silica
gel to provide the 5'-ethynyl compound of formula (5).
Step 5--Preparation of Formula I
[0098] The acetonide-protected compound of formula (5) is then
converted into a compound of Formula I in which Y is a covalent
bond, Z is --C.ident.C--, and R.sup.3 is hydrogen by treatment with
an acid, for example an organic acid, for example acetic acid. The
reaction is carried out in a mixture of the acid and water, at
about 50-100.degree. C., preferably about 80-90.degree. C., for
about 10-48 hours, preferably about 16 hours. When the reaction is
substantially complete, the product of Formula I is isolated by
conventional means, for example by removal of the solvent under
reduced pressure, followed by chromatography of the residue on
silica gel.
Alternative Preparation of a Compound of Formula I
[0099] The compounds of Formula I where R.sup.3 is hydrogen,
X.sup.1 and Y are covalent bonds, and Z is --C.ident.C-- may
alternatively be prepared starting from a compound of formula (1)
as shown in Reaction Scheme IA. This method of synthesis is
preferred when there is substitution on the R.sup.1 moiety, for
example a hydroxy substituent. ##STR5## ##STR6## Step 1 is carried
out as shown in Reaction Scheme I. Step 2--Preparation of Formula
(4a)
[0100] The hydroxymethyl compound of formula (2) is oxidized to an
aldehyde of formula (4a) using a modification of the Moffat
Oxidation. In general, to the compound of formula (2) is added a
mixture of DCC, dimethysulfoxide and pyridine. The initial reaction
is carried out at a temperature of about -5.degree. to about
10.degree. C., preferably about 0.degree. C., and then at about
room temperature for about 6-48 hours, preferably about 18 hours.
When the reaction is substantially complete, the aldehyde of
formula (4a) is isolated by conventional means, for example by
partitioning the product between ethyl acetate and water and
removing the solvent under reduced pressure. The product is used in
the next step without further purification.
Step 3--Preparation of Formula (5a)
[0101] The 5'-aldehyde group is converted to an ethynyl group by
reaction with bromomethyltriphenylphosphonium bromide in the
presence of a strong base, preferably potassium t-butoxide. The
reaction is carried out in an inert solvent, preferably
tetrahydrofuran, at a temperature of about -80.degree. C., allowing
the reaction mixture to gradually warm to room temperature over a
period of about 1-3 days. When the reaction is substantially
complete, the product of formula (5a) is isolated by conventional
means.
Step 4--Preparation of Formula (5)
[0102] The 6-chloro moiety is displaced from the compound of
formula (5a) by reaction with a compound of formula
R.sup.1XNH.sub.2, where X is as defined above, in the presence of a
base, for example triethylamine. The reaction is carried out in an
inert protic solvent, for example ethanol, at a temperature of
about reflux, for about 14-48 hours, preferably about 16 hours.
When the reaction is substantially complete, the product of Formula
(5) is isolated by conventional means.
Step 5--Preparation of Formula I
[0103] The compound of formula (5) is then converted to a compound
of Formula I as shown in Reaction Scheme I above.
Preparation of a Compound of Formula I in which R.sup.3 is not
Hydrogen
[0104] The preparation of a compound of Formula I in which R.sup.3
is not hydrogen, X.sup.1 is a covalent bond, Y is as defined above,
and Z is --C.ident.C-- is shown in Reaction Scheme II. ##STR7##
Step 1--Preparation of Formula (6)
[0105] The 4'-ethynyl compound of formula (5) is converted to a
compound of formula (6) by reaction with a compound of the formula
R 3Y-LG, in which LG is a leaving group, preferably a halogen, for
example iodo or bromo. The reaction is carried out in the presence
of catalytic amounts of
dichlorobis(triphenylphosphine)palladium(II) and copper(II)iodide
plus a tertiary amine, for example triethylamine, in an inert
solvent, for example tetrahydrofuran, at a temperature of about
room temperature for about 15 minutes. When the reaction is
substantially complete, the product of Formula I is isolated by
conventional means, for example by removal of the solvent under
reduced pressure, followed by preparative chromatography on silica
gel to provide the 5'-substituted ethynyl compound of Formula
I.
Step 2--Preparation of Formula I
[0106] The compound of formula (6) is then deprotected in the same
manner as shown above in Reaction Scheme 1 by treatment with an
acid, preferably an organic acid, for example acetic acid, to
provide a compound of Formula I.
Alternative Preparation of a Compound of Formula I in which R.sup.3
is not Hydrogen
[0107] Alternatively, a compound of Formula I in which R.sup.3 is
not hydrogen, X.sup.1 is a covalent bond, Y is as defined above,
and Z is --C.ident.C-- may be prepared directly from a compound of
Formula I in which R.sup.3 is hydrogen as shown in Reaction Scheme
IIA. ##STR8## Step 1
[0108] The 4'-ethynyl compound of Formula I is converted to a
compound of Formula I in which R.sup.3 is not hydrogen by reaction
with a compound of the formula R.sup.3Y-LG, in which LG is a
leaving group, preferably a halogen, for example iodo or bromo. The
reaction is carried out in the presence of catalytic amounts of
dichlorobis(triphenylphosphine)palladium(II) and copper(I)iodide
plus a tertiary amine, for example triethylamine, in an inert
solvent, for example tetrahydrofuran, at a temperature of about
room temperature for about 15 minutes. When the reaction is
substantially complete, the product of Formula I in which R.sup.3
is not hydrogen is isolated by conventional means, for example by
removal of the solvent under reduced pressure, followed by
preparative chromatography on silica gel to provide the
4'-substituted ethynyl compound of Formula I.
Preparation of a Compound of Formula I in which Z is
--CH.dbd.CH--
[0109] The preparation of a compound of Formula I in which X.sup.1
is a covalent bond and Z is --CH.dbd.CH-- is shown in Reaction
Scheme III. ##STR9## Step 1--Preparation of Formula (7)
[0110] The 4'-aldehyde group is converted to an ethenyl group using
the Wittig reaction, by reaction of a compound of formula (4), the
preparation of which was shown above, with
R.sup.3Y--CH.sub.2P(Ph).sub.3Br (where Ph is phenyl), in the
presence of a base, for example aqueous sodium-hydroxide. The
reaction is carried out in an inert solvent, for example
dichloromethane, at a temperature of about room temperature, over a
period of about 1-10 hours. When the reaction is substantially
complete, the product of formula (7) is isolated by conventional
means, for example by removal of the solvent under reduced
pressure, followed by partition between a solvent such as ethyl
acetate and water, removing the solvent under reduced pressure. The
residue may be further purified by chromatography on silica gel to
provide the 4'-ethenyl compound of formula (7).
Step 2--Preparation of a Compound of Formula I in which Z is
--CH.dbd.CH--
[0111] The compound of formula (7) is then deprotected in the same
manner as shown above in Reaction Scheme 1 by treatment with an
acid, for example an organic acid, for example acetic acid, to
provide a compound of Formula I.
Preparation of a Compound of Formula I in which Z is
--CH.sub.2CH.sub.2--
[0112] The preparation of a compound of Formula I in which X.sup.1
is a covalent bond and Z is --CH.sub.2CH.sub.2-- is shown in
Reaction Scheme IV. ##STR10##
[0113] A compound of Formula I in which Z is --CH.sub.2CH.sub.2--
is prepared from a compound of Formula I in which Z is
--CH.dbd.CH-- (or, alternatively, a compound of Formula I in which
Z is --C.ident.C-- may be used). In general, the compound of
Formula I in which Z is --CH.dbd.CH-- is dissolved in an inert
solvent and stirred with a catalyst, for example palladium
hydroxide, and a catalytic hydrogenation transfer reagent, such as
cyclohexene. The reaction is carried out in an inert solvent, for
example ethanol, at about room temperature, over a period of about
10-48 hours. When the reaction is substantially complete, the
product of Formula I is isolated by conventional means, for example
by removal of the solvent under reduced pressure, followed by
chromatography, to provide a compound of Formula I in which Z is
--CH.sub.2CH.sub.2--.
B. Preparation of Formula I where R.sup.2 is Hydrogen and X.sup.1
is CH.sub.2.
[0114] The compounds of Formula I where R.sup.3 is hydrogen, Y is a
covalent bond, X.sup.1 is CH.sub.2, and Z is --C.ident.C-- may be
prepared starting from a compound of formula (4), as shown in
Reaction Scheme V. ##STR11## ##STR12## Step 1--Preparation of
Formula (8)
[0115] N-sodium hexamethyldisilazane is reacted with
benzyloxymethyltriphenylphosphonium chloride (formula 4a) at a
temperature of about -80.degree. C., for about 1 hour. The compound
of formula (4), prepared as shown in Reaction Scheme 1, or by means
well known to those in the art, is dissolved in an inert solvent,
for example tetrahydrofuran, and added to the reaction mixture,
which is allowed to rise to about room temperature, and stirred
from 4-24 hours preferably 8 hours. When the reaction is
substantially complete, the product is isolated and purified
conventionally, for example by crystallization of the residue.
Step 2a. Preparation of Formula (9)
[0116] The compound of formula (9) is prepared conventionally from
the compound of formula (8) by hydrogenation in the presence of
with a catalyst, for example Pd/C. When the reaction is
substantially complete, the product of formula (9) is isolated by
conventional means and used without further purification.
Step 2b. Preparation of Formula (10)
[0117] The compound of formula (9) is deprotected by hydrogenation
in the presence of a catalyst, for example palladium hydroxide, and
a catalytic hydrogenation transfer reagent, such as cyclohexene.
The reaction is carried out in an inert solvent, for example
ethanol, at about room temperature, over a period of about 2-7
days, preferably 5 days, at about 75-100.degree. C., preferably
about 80.degree. C. When the reaction is substantially complete,
the product of formula (10) is isolated by conventional means, for
example by removal of the solvent by filtration, followed by
purification by chromatography on silica gel, to provide the
compound of formula (10).
Step 3.--Preparation of Formula (11)
[0118] The hydroxymethyl compound of formula (10) is oxidized to an
aldehyde of formula (11) using a modification of the Moffat
Oxidation. In general, the compound of formula (10) is reacted with
a mixture of dicyclohexylcarbodiimide, dimethylsulfoxide and
pyridine. The initial reaction is carried out at a temperature of
about --5.degree.-10.degree. C., preferably about 0.degree. C., and
then at about room temperature for about 6-48 hours, preferably
about 18 hours. When the reaction is substantially complete, the
aldehyde of formula (11) is isolated by conventional means. The
product is for example used in the next step without further
purification.
Step 4--Preparation of Formula (12)
[0119] The 4'-aldehyde group is converted to an ethynyl group by
reaction with bromomethyltriphenylphosphonium bromide in the
presence of a strong base, for example potassium t-butoxide. The
reaction is carried out in an inert solvent, for example
tetrahydrofuran, at a temperature of about -80.degree. C., allowing
the reaction mixture to gradually warm to room temperature, and
stirring for about 1-3 days. When the reaction is substantially
complete, the product of formula (12) is isolated and purified by
conventional means, for example by chromatography on silica gel, to
provide the 5'-ethynyl compound of formula (12).
Step 5--Preparation of Formula I
[0120] The acetonide-protected compound of formula (12) is then
converted into a compound of Formula I in which Y is a covalent
bond, Z is --C.ident.C--, and R.sup.3 is hydrogen by treatment with
an acid, for example acetic acid. The reaction is carried out in a
mixture of the acid and water, at about 50-100.degree. C.,
preferably about 80-90.degree. C., for about 10-48 hours,
preferably about 16 hours. When the reaction is substantially
complete, the product of Formula I is isolated and purified by
conventional means, for example by chromatography of the residue on
silica gel. Preparation of Compounds of Formula I where X.sup.1 is
(CH.sub.2).sub.2. ##STR13##
[0121] Compounds of Formula (I) where X.sup.1 is (CH.sub.2).sub.2
are obtained as shown in Reaction Scheme V but replacing the
compound of Formula (4a) with a compound of Formula (4b). ##STR14##
Synthesis of Compounds (4a) and (4b)
[0122] Chloro(phenylmethoxy)ethane and triphenylphosphine are
reacted in an inert solvent, for example benzene, and maintained
under reflux conditions overnight. When the reaction is
substantially complete, the product of formula (4a) is isolated
conventionally.
[0123] Similarly, by replacing chloro(phenylmethoxy) methane with
chloro(phenylmethoxy) ethane, a compound of formula (4b) is
prepared.
[0124] Compounds of Formula I where X.sup.1 is (CH.sub.2).sub.2 and
R.sup.3 is hydrogen are converted to compounds of Formula I where
X.sup.1 is (CH.sub.2).sub.2 and R.sup.3 is other than hydrogen as
shown in Reaction Scheme II above.
Utility, Testing and Administration
General Utility
[0125] The compounds of Formula I are effective in the treatment of
conditions known to respond to administration of a partial or full
agonist of an A.sub.1 adenosine receptor. Such conditions include,
but are not limited to, acute and chronic disorders of heart
rhythm, especially those diseases characterized by rapid heart
rate, in which the rate is driven by abnormalities in the
sinoatrial, atria, and AV nodal tissues. Such disorders include,
but are not limited to, atrial fibrillation, supraventricular
tachycardia and atrial flutter, congestive heart failure and sudden
death resulting from arrythmia, non-insulin-dependent diabetes
mellitus, hyperglycemia, epilepsy (anticonvulsant activity), and
cardio-and neuro- protection.
[0126] A.sub.1 agonists, as a result of their inhibitory action on
cyclic AMP generation, have antilipolytic effects in adipocytes
that lead to a decreased release of nonesterified fatty acids
(NEFA) (E. A. van Schaick et al J. Pharmacokinetics and
Biopharmaceutics, Vol. 25 (1997) p 673-694 and P. Strong Clinical
Science Vol. 84 (1993) p. 663-669). Non-insulin-dependent diabetes
mellitus (NIDDM) is characterized by an insulin resistance that
results in hyperglycemia. Factors contributing to the observed
hyperglycemia are a lack of normal glucose uptake and activation of
skeletal muscle glycogen synthase (GS). Elevated levels of NEFA
have been shown to inhibit insulin-stimulated glucose uptake and
glycogen synthesis (D. Thiebaud et al Metab. Clin. Exp. Vol. 31
(1982) p 1128-1136 and G. Boden et al J. Clin. Invest. Vol. 93
(1994) p 2438-2446). The hypothesis of a glucose fatty acid cycle
was proposed by P. J. Randle as early as 1963 (P. J. Randle et al
Lancet (1963) p. 785-789). Thus, limiting the supply of fatty acids
to the peripheral tissues promotes carbohydrate utilization (P.
Strong et al Clinical Science Vol. 84 (1993) p. 663-669).
[0127] The benefit of an A.sub.1 agonist in central nervous
disorders has been reviewed (L. J. S. Knutsen and T. F. Murray In
Purinergic Approaches in Experimental Therapeutics, Eds. K. A.
Jacobson and M. F. Jarvis (1997) Wiley-Liss, N.Y., P--423-470).
Briefly, based on experimental models of epilepsy, a mixed
A.sub.2A:A.sub.1 agonist, metrifudil, has been shown to be a potent
anticonvulsant against seizures induced by the inverse
benzodiazepine agonist methyl
6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM, H.
Klitgaard Eur. J. Pharmacol. (1993) Vol. 224 p. 221-228). In other
studies using CGS 21680, an A.sub.2A agonist, it was concluded that
the anticonvulsant activity was attributed to activation of the
A.sub.1 receptor (G. Zhang et al. Eur. J. Pharmacol. Vol. 255
(1994) p. 239-243). Furthermore, A.sub.1 adenosine selective
agonists have been shown to have anticonvulsant activity in the
DMCM model (L. J. S. Knutsen In Adenosine and Adenne Nucleotides:
From Molecular Biology to Integrative Physiology; eds. L.
Belardinelli and A. Pelleg, Kluwer: Boston, 1995, pp 479-487). A
second area where an A.sub.1 adenosine agonist has a benefit is in
animal models of forebrain ishemia as demonstrated by Knutsen et al
(J. Med. Chem. Vol. 42 (1999) p. 3463-3477). The benefit in
neuroprotection is believed to be in part due to the inhibition of
the release of excitatory amino acids (ibid).
Testing
[0128] Activity testing is conducted as described in those patents
and literature citations referenced above, and in the Examples
below, and by methods apparent to one skilled in the art.
Pharmaceutical Compositions
[0129] The compounds of Formula I are usually administered in the
form of pharmaceutical compositions. This invention therefore
provides pharmaceutical compositions that contain, as the active
ingredient, one or more of the compounds of Formula I, or a
pharmaceutically acceptable salt or ester thereof, and one or more
pharmaceutically acceptable excipients, carriers, including inert
solid diluents and fillers, diluents, including sterile aqueous
solution and various organic solvents, permeation enhancers,
solubilizers and adjuvants. The compounds of Formula I may be
administered alone or in combination with other therapeutic agents.
Such compositions are prepared in a manner well known in the
pharmaceutical art (see, e.g., Remington's Pharmaceutical Sciences,
Mace Publishing Co., Philadelphia, Pa. 17.sup.th Ed. (1985) and
"Modern Pharmaceutics", Marcel Dekker, Inc. 3.sup.rd Ed. (G. S.
Banker & C. T. Rhodes, Eds.).
Administration
[0130] The compounds of Formula I may be administered in either
single or multiple doses by any of the accepted modes of
administration of agents having similar utilities, for example as
described in those patents and patent applications incorporated by
reference, including rectal, buccal, intranasal and transdermal
routes, by intra-arterial injection, intravenously,
intraperitoneally, parenterally, intramuscularly, subcutaneously,
orally, topically, as an inhalant, or via an impregnated or coated
device such as a stent, for example, or an artery-inserted
cylindrical polymer.
[0131] One mode for administration is parental, particularly by
injection. The forms in which the novel compositions of the present
invention may be incorporated for administration by injection
include aqueous or oil suspensions, or emulsions, with sesame oil,
corn oil, cottonseed oil, or peanut oil, as well as elixirs,
mannitol, dextrose, or a sterile aqueous solution, and similar
pharmaceutical vehicles. Aqueous solutions in saline are also
conventionally used for injection, but less preferred in the
context of the present invention. Ethanol, glycerol, propylene
glycol, liquid polyethylene glycol, and the like (and suitable
mixtures thereof), cyclodextrin derivatives, and vegetable oils may
also be employed. The proper fluidity can be maintained, for
example, by the use of a coating, such as lecithin, by the
maintenance of the required particle size in the case of dispersion
and by the use of surfactants. The prevention of the action of
microorganisms can be brought about by various antibacterial and
antifungal agents, for example, parabens, chlorobutanol, phenol,
sorbic acid, thimerosal, and the like.
[0132] Sterile injectable solutions are prepared by incorporating
the compound of Formula I in the required amount in the appropriate
solvent with various other ingredients as enumerated above, as
required, followed by filtered sterilization. Generally,
dispersions are prepared by incorporating the various sterilized
active ingredients into a sterile vehicle which contains the basic
dispersion medium and the required other ingredients from those
enumerated above. In the case of sterile powders for the
preparation of sterile injectable solutions, the preferred methods
of preparation are vacuum-drying and freeze-drying techniques which
yield a powder of the active ingredient plus any additional desired
ingredient from a previously sterile-filtered solution thereof.
[0133] Oral administration is another route for administration of
the compounds of Formula I. Administration may be via capsule or
enteric coated tablets, or the like. In making the pharmaceutical
compositions that include at least one compound of Formula I, the
active ingredient is usually diluted by an excipient and/or
enclosed within such a carrier that can be in the form of a
capsule, sachet, paper or other container. When the excipient
serves as a diluent, in can be a solid, semi-solid, or liquid
material (as above), which acts as a vehicle, carrier or medium for
the active ingredient. Thus, the compositions can be in the form of
tablets, pills, powders, lozenges, sachets, cachets, elixirs,
suspensions, emulsions, solutions, syrups, aerosols (as a solid or
in a liquid medium), ointments containing, for example, up to 10%
by weight of the active compound, soft and hard gelatin capsules,
sterile injectable solutions, and sterile packaged powders.
[0134] Some examples of suitable excipients include lactose,
dextrose, sucrose, sorbitol, mannitol, starches, gum acacia,
calcium phosphate, alginates, tragacanth, gelatin, calcium
silicate, microcrystalline cellulose, polyvinylpyrrolidone,
cellulose, sterile water, syrup, and methyl cellulose. The
formulations can additionally include: lubricating agents such as
talc, magnesium stearate, and mineral oil; wetting agents;
emulsifying and suspending agents; preserving agents such as
methyl- and propylhydroxy-benzoates; sweetening agents; and
flavoring agents.
[0135] The compositions of the invention can be formulated so as to
provide quick, sustained or delayed release of the active
ingredient after administration to the patient by employing
procedures known in the art. Controlled release drug delivery
systems for oral administration include osmotic pump systems and
dissolutional systems containing polymer-coated reservoirs or
drug-polymer matrix formulations. Examples of controlled release
systems are given in U.S. Pat. Nos. 3,845,770; 4,326,525; 4,902514;
and 5,616,345. Another formulation for use in the methods of the
present invention employs transdermal delivery devices ("patches").
Such transdermal patches may be used to provide continuous or
discontinuous infusion of the compounds of the present invention in
controlled amounts. The construction and use of transdermal patches
for the delivery of pharmaceutical agents is well known in the art.
See, e.g., U.S. Pat. Nos. 5,023,252, 4,992,445 and 5,001,139. Such
patches may be constructed for continuous, pulsatile, or on demand
delivery of pharmaceutical agents.
[0136] The compositions are formulated in a unit dosage form. The
term "unit dosage forms" refers to physically discrete units
suitable as unitary dosages for human subjects and other mammals,
each unit containing a predetermined quantity of active material
calculated to produce the desired therapeutic effect, in
association with a suitable pharmaceutical excipient (e.g., a
tablet, capsule, ampoule). The compounds of Formula I are effective
over a wide dosage range and is generally administered in a
pharmaceutically effective amount. For example, for oral
administration, each dosage unit contains from 10 mg to 2 g of a
compound of Formula I, more preferably from 10 to 700 mg, and for
parenteral administration, preferably from 10 to 700 mg of a
compound of Formula I, more preferably about 50-200 mg. It will be
understood, however, that the amount of the compound of Formula I
actually administered will be determined by a physician, in the
light of the relevant circumstances, including the condition to be
treated, the chosen route of administration, the actual compound
administered and its relative activity, the age, weight, and
response of the individual patient, the severity of the patient's
symptoms, and the like.
[0137] For preparing solid compositions such as tablets, the
principal active ingredient is mixed with a pharmaceutical
excipient to form a solid preformulation composition containing a
homogeneous mixture of a compound of the present invention. When
referring to these preformulation compositions as homogeneous, it
is meant that the active ingredient is dispersed evenly throughout
the composition so that the composition may be readily subdivided
into equally effective unit dosage forms such as tablets, pills and
capsules.
[0138] The tablets or pills of the present invention may be coated
or otherwise compounded to provide a dosage form affording the
advantage of prolonged action, or to protect from the acid
conditions of the stomach. For example, the tablet or pill can
comprise an inner dosage and an outer dosage component, the latter
being in the form of an envelope over the former. The two
components can be separated by an enteric layer that serves to
resist disintegration in the stomach and permit the inner component
to pass intact into the duodenum or to be delayed in release. A
variety of materials can be used for such enteric layers or
coatings, such materials including a number of polymeric acids and
mixtures of polymeric acids with such materials as shellac, cetyl
alcohol, and cellulose acetate.
[0139] Compositions for inhalation or insufflation include
solutions and suspensions in pharmaceutically acceptable, aqueous
or organic solvents, or mixtures thereof, and powders. The liquid
or solid compositions may contain suitable pharmaceutically
acceptable excipients as described supra. The compositions are
administered by the oral or nasal respiratory route for local or
systemic effect. Compositions in pharmaceutically acceptable
solvents may be nebulized by use of inert gases. Nebulized
solutions may be inhaled directly from the nebulizing device or the
nebulizing device may be attached to a face mask tent, or
intermittent positive pressure breathing machine. Solution,
suspension, or powder compositions may be administered, for example
orally or nasally, from devices that deliver the formulation in an
appropriate manner.
[0140] The following examples are included to demonstrate preferred
embodiments of the invention. It should be appreciated by those of
skill in the art that the techniques disclosed in the examples
which follow represent techniques discovered by the inventor to
function well in the practice of the invention, and thus can be
considered to constitute preferred modes for its practice. However,
those of skill in the art should, in light of the present
disclosure, appreciate that many changes can be made in the
specific embodiments which are disclosed and still obtain a like or
similar result without departing from the spirit and scope of the
invention.
EXAMPLE 1
Preparation of a Compound of Formula (3)
A. Preparation of a Compound of Formula (3) where R.sup.1 is
Cyclopentyl R.sup.2 is Hydrogen and X is a Covalent Bond
[0141] ##STR15##
[0142] To a solution of
(2S,1R,4R,5R)-2-hydroxymethyl-5-(6-chloropurin-9-yl)-tetrahydrofuran-3,4--
diol acetonide, the compound of formula (2) in which R.sup.2 is
hydrogen (4.98 g, 15 mmol) in ethanol (80 ml) was added
cyclopentylamine (0.6 ml, 30 mmol), and triethylamine (6.27 ml, 45
mmol), and the mixture was refluxed for 16 hours. The solvent was
then removed under reduced pressure, and the residue partitioned
between ethyl acetate and 10% citric acid in water, followed by
water. Ethyl acetate was removed from the organic layer, to yield
{(1R,2R,4R,5R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}methan-1-ol, a compound of formula
(3).
B. Preparation of Compounds of Formula (3) Varying X, R.sup.1 and
R.sup.2
[0143] Similarly, following the procedure of 1A above, but
replacing cyclopentylamine with other amines of formula
R.sup.1XN.sub.2, the following compound of formula (3) was
prepared:
{(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxa-4-[6-(oxolan-3-ylamino)purin-9-y-
l]bicyclo[3.3.0]oct-2-yl}methan-1-ol.
C. Preparation of Compounds of Formula (3), Varying X, R.sup.1 and
R.sup.2
[0144] Similarly, following the procedure of 1A above, but
replacing cyclopentylamine with other amines of formula
R.sup.1XNH.sub.2, the following compounds of formula (3) are
prepared: [0145]
{(1R,2R,4R,5R)-4-[6-(cyclopentylmethylamino)purin-9-yl]-7,7-dimethyl-3,6,-
8-trioxabicyclo[3.3.0]oct-2-yl}methan-1-ol; [0146]
{(1R,2R,4R,5R)-4-[2-trifluoromethyl-6-(cyclopentylamino)purin-9-yl]-7,7-d-
imethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl}methan-1-ol; [0147]
{(1R,2R,4R,5R)-4-[6-cyclobutylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trioxa-
bicyclo[3.3.0]oct-2-yl}methan-1-ol; [0148]
{(1R,2R,4R,5R)-4-[6-cyclohexylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trioxa-
bicyclo[3.3.0]oct-2-yl}methan-1-ol; [0149]
{(1R,2R,4R,5R)-4-[2-fluoro-6-cyclohexylamino)purin-9-yl]-7,7-dimethyl-3,6-
,8-trioxabicyclo[3.3.0]oct-2-yl}methan-1-ol; [0150]
{(1R,2R,4R,5R)-4-[6-cyclohexylmethylamino)purin-9-yl]-7,7-dimethyl-3,6,8--
trioxabicyclo[3.3.0]oct-2-yl}methan-1-ol; [0151]
{(1R,2R,4R,5R)-4-[6-(3-fluorocyclopentylamino)purin-9-yl]-7,7-dimethyl-3,-
6,8-trioxabicyclo[3.3.0]oct-2-yl}methan-1-ol; [0152]
{(1R,2R,4R,5R)-4-[6-(4-trifluoromethylcyclopentylamino)purin-9-yl]-7,7-di-
methyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl}methan-1-ol; [0153]
{(1R,2R,4R,5R)-4-[6-(3-methoxycyclopentylamino)purin-9-yl]-7,7-dimethyl-3-
,6,8-trioxabicyclo[3.3.0]oct-2-yl}methan-1-ol; [0154]
{(1R,2R,4R,5R)-4-[6-(phenylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}methan-1-ol; [0155]
{(1R,2R,4R,5R)-4-[6-(benzylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2yl}methan-1-ol; [0156]
{(1R,2R,4R,5R)-4-[6-(4-fluorophenylamino)purin-9-yl]-7,7-dimethyl-3,6,8-t-
rioxabicyclo[3.3.0]oct-2-yl}methan-1-ol; [0157]
{(1R,2R,4R,5R)-4-[6-(pyridin-3-ylamino)purin-9-yl]-7,7-dimethyl-3,6,8-tri-
oxabicyclo[3.3.0]oct-2-yl}methan-1-ol; [0158]
{(1R,2R,4R,5R)-4-[6-(thiazol-2-ylamino)purin-9-yl]-7,7-dimethyl-3,6,8-tri-
oxabicyclo[3.3.0]oct-2-yl}methan-1-ol; [0159]
{(1R,2R,4R,5R)-4-[6-(tetrahydropyran-3-ylamino)purin-9-yl]-7,7-dimethyl-3-
,6,8-trioxabicyclo[3.3.0]oct-2-yl}methan-1-ol; [0160]
{(1R,2R,4R,5R)-4-[6-(tetrahydropyran-3-ylmethylamino)purin-9-yl]-7,7-dime-
thyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl}methan-1-ol; and [0161]
{(1R,2R,4R,5R)-4-[6-(5-fluorotetrahydropyran-3-ylamino)purin-9-yl]-7,7-di-
methyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl}methan-1-ol.
D. Preparation of Compounds of Formula (3), Varying X, R.sup.1 and
R.sup.2
[0162] Similarly, following the procedure of 1A above, but
replacing cyclopentylamine with other amines of formula
R.sup.1XNH.sub.2, other compounds of formula (3) are prepared.
EXAMPLE 2
Preparation of a Compound of Formula (4)
A. Preparation of a Compound of Formula (4) where R.sup.1 is
Cyclopentyl, R.sup.2 is Hydrogen, and X is a Covalent Bond
[0163] ##STR16##
[0164] A mixture of
{(1R,2R,4R,5R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}methan-1-ol (0.94 g, 25 mmol),
dimethylsulfoxide (7 ml), dicyclohexylcarbodimide (1.55 g) and
pyridine (0.2 ml) was stirred at 0.degree. C. for a few minutes,
and then trifluoroacetic acid (0.1 ml) added. The mixture was
allowed to warn to room temperature, and stirred for 18 hours. The
mixture was then partitioned between ethyl acetate and water and
washed with water. Solvent was removed from the organic layer under
reduced pressure, and the product,
(2S,1R,4R,5R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-triox-
abicyclo[3.3.0]octane-2-carbaldehyde, a compound of formula
(4).
B. Preparation of Compounds of Formula (4), Varying R.sup.1
[0165] Similarly, following the procedure of 2A above, but
replacing
{(1R,2R,4R,5R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}methan-1-ol with
{(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxa-4-[6-(oxolan-3-ylamino)purin-9-y-
l]bicyclo[3.3.0]oct-2-yl}methan-1-ol, the following compound of
formula (4) was prepared: [0166]
(2S,1R,4R,5R)-7,7-dimethyl-3,6,8-trioxa-4-[6-(oxolan-3-ylamino)purin-9-yl-
]bicyclo[3.3.0]octane-2-carboxaldehyde.
C. Preparation of Compounds of Formula (4), Varying X, R.sup.1 and
R.sup.2
[0167] Similarly, following the procedure of 2A above, but
replacing
{(1R,2R,4R,5R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}methan-1-ol with other compounds of
formula (3), the following compounds of formula (4) are prepared:
[0168]
{(1R,2R,4R,5R)-4-[6-(cyclopentylmethylamino)purin-9-yl]-7,7-dimethyl-3,6,-
8-trioxabicyclo[3.3.0]octane-2-carboxaldehyde; [0169]
{(1R,2R,4R,5R)-4-[2-trifluoromethyl-6-(cyclopentylamino)purin-9-yl]-7,7-d-
imethyl-3,6,8-trioxabicyclo[3.3.0]octane-2-carboxaldehyde; [0170]
{(1R,2R,4R,5R)-4-[6-cyclobutylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trioxa-
bicyclo[3.3.0]octane-2-carboxaldehyde; [0171]
{(1R,2R,4R,5R)-4-[6-cyclohexylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trioxa-
bicyclo[3.3.0]octane-2-carboxaldehyde; [0172]
{(1R,2R,4R,5R)-4-[2-fluoro-6-cyclohexylamino)purin-9-yl]-7,7-dimethyl-3,6-
,8-trioxabicyclo[3.3.0]octane-2-carboxaldehyde; [0173]
{(1R,2R,4R,5R)-4-[6-cyclohexylmethylamino)purin-9-yl]-7,7-dimethyl-3,6,8--
trioxabicyclo[3.3.0]octane-2-carboxaldehyde; [0174]
{(1R,2R,4R,5R)-4-[6-(3-fluorocyclopentylamino)purin-9-yl]-7,7-dimethyl-3,-
6,8-trioxabicyclo[3.3.0]octane-2-carboxaldehyde; [0175]
{(1R,2R,4R,5R)-4-[6-(4-trifluoromethylcyclopentylamino)purin-9-yl]-7,7-di-
methyl-3.6.8-trioxabicyclo[3.3.0]octane-2-carboxaldehyde; [0176]
{(1R,2R,4R,5R)-4-[6-(3-methoxycyclopentylamino)purin-9-yl]-7,7-dimethyl-3-
,6,8-trioxabicyclo[3.3.0]octane-2-carboxaldehyde; [0177]
{(1R,2R,4R,5R)-4-[6-(phenylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]octane-2-carboxaldehyde; [0178]
{(1R,2R,4R,5R)-4-[6-(benzylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]octane-2-carboxaldehyde; [0179]
{(1R,2R,4R,5R)-4-[6-(4-fluorophenylamino)purin-9-yl]-7,7-dimethyl-3,6,8-t-
rioxabicyclo[3.3.0]octane-2-carboxaldehyde; [0180]
{(1R,2R,4R,5R)-4-[6-(pyridin-3-ylamino)purin-9-yl]-7,7-dimethyl-3,6,8-tri-
oxabicyclo[3.3.0]octane-2-carboxaldehyde; [0181]
{(1R,2R,4R,5R)-4-[6-(thiazol-2-ylamino)purin-9-yl]-7,7-dimethyl-3,6,8-tri-
oxabicyclo[3.3.0]octane-2-carboxaldehyde; [0182]
{(1R,2R,4R,5R)-4-[6-(tetrahydropyran-3-ylamino)purin-9-yl]-7,7-dimethyl-3-
,6,8-trioxabicyclo[3.3.0]octane-2-carboxaldehyde; [0183]
{(1R,2R,4R,5R)-4-[6-(tetrahydropyran-3-ylmethylamino)purin-9-yl]-7,7-dime-
thyl-3,6,8-trioxabicyclo[3.3.0]octane-2-carboxaldehyde; and [0184]
{(1R,2R,4R,5R)-4-[6-(5-fluorotetrahydropyran-3-ylamino)purin-9-yl]-7,7-di-
methyl-3,6,8-trioxabicyclo[3.3.0]octane-2-carboxaldehyde.
D. Preparation of Compounds of Formula (4), Varying X, R.sup.1 and
R.sup.2
[0185] Similarly, following the procedure of 2A above, but
replacing
{(1R,2R,4R,5R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl) methan-1-ol with other compounds of
formula (3), other compounds of formula (4) are prepared.
EXAMPLE 3
Preparation of a Compound of Formula (5)
A. Preparation of a Compound of Formula (5) where R.sup.1 is
Cyclopentyl and R.sup.2 is Hydrogen
[0186] ##STR17##
[0187] To a suspension of potassium t-butoxide (0.84 g, 7.5 mmol)
in tetrahydrofuran (5 ml) at -78.degree. C. was added
bromomethyltriphenylphosphonium (1.64 g, 3.75 mmol) in small
portions, and the mixture stirred for 2 hours. To this mixture was
added a solution of
(2S,1R,4R,5R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-tr-
ioxabicyclo[3.3.0]octane-2-carboxaldehyde (0.932 g, 2.5 mmol) in
tetrahydrofuran (20 ml), and the mixture was stirred for 2 hours at
-78.degree. C. The reaction mixture was then allowed to warm to
room temperature and stirred for 6 days, then quenched with aqueous
ammonium chloride, and partitioned between water and ethyl acetate.
The organic layer was separated, dried over magnesium sulfate,
filtered, and the solvent removed from the filtrate under reduced
pressure, to yield
[9-((1R,2S,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2--
yl)purin-9-yl]cyclopentylamine, a compound of formula (5).
B. Preparation of Compounds of Formula (5), Varying R.sup.1
[0188] Similarly, following the procedure of 3A above, but
replacing
(2S,1R,4R,5R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-triox-
abicyclo[3.3.0]octane-2-carboxaldehyde with
{(1R,2S,4R,5R)-7,7-dimethyl-3,6,8-trioxa-4-[6-(oxolan-3-ylamino)purin-9-y-
l]bicyclo[3.3.0]octane-2-carboxaldehyde, the following compound of
formula (5) was prepared:
(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxa
bicyclo[3.3.0]oct-2-yl)-purin-9-yl]oxolan-3-ylamine.
C. Preparation of Compounds of Formula (4), Varying X, R.sup.1 and
R.sup.2
[0189] Similarly, following the procedure of 3A above, but
replacing
{(1R,2R,4R,5R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}methan-1-ol with other compounds of
formula (3), the following compounds of formula (4) are prepared:
[0190]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl]cyclopentylmethylamine; [0191]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-2-trifluoromethyl-3,6,8-trioxabicyc-
lo[3.3.0]oct-2-yl-purin-9-yl]cyclopentylamine; [0192]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl]cyclobutylamine; [0193]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl]cyclohexylamine; [0194]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl]2-fluoro-6-cyclohexylamine; [0195]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl]cyclohexylmethylamine; [0196]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl](3-fluorocyclopentylamine; [0197]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl](4-trifluoromethylcyclopentylamine; [0198]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl](3-methoxycyclopentylamine; [0199]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl](phenylamine; [0200]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-y-p-
urin-9-yl]benzylamine; [0201]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl](4-fluorophenylamine); [0202]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl](pyridin-3-ylamine; [0203]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-ylp-
urin-9-yl](thiazol-2-ylamine; [0204]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl](tetrahydropyran-3-ylamine); [0205]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl](tetrahydropyran-3-ylmethylamine; and [0206]
{(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl--
purin-9-yl](5-fluorotetrahydropyran-3-ylamine.
D. Preparation of Compounds of Formula (5), Varying X, R.sup.1 and
R.sup.2
[0207] Similarly, following the procedure of 3A above, but
replacing
{(1R,2R,4R,5R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}methan-1-ol with other compounds of
formula (4), other compounds of formula (5) are prepared.
EXAMPLE 4
Preparation of a Compound of Formula I
A. Preparation of a Compound of Formula I where R.sup.1 is
Cyclopentyl, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are Hydrogen, X,
X.sup.1 and Y are Covalent Bonds, and Z is --C.ident.C--
[0208] ##STR18##
[0209] A solution of
[9-((1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2y-
l)purin-6-yl]cyclopentylamine (0.28 g) was dissolved in 20 ml of a
mixture of acetic acid:water (80:20) and stirred overnight at
75.degree. C. Solvent was removed under reduced pressure, and the
residue purified by preparative TLC, eluting with
methanol:methylene chloride (1:8), to yield
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-ethynyloxolane-3,4-dio-
l, a compound of Formula I.
B. Preparation of a Compound of Formula I where R.sup.1 is
Tetrahydrofuran-3-yl, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are
Hydrogen, X, X.sup.1 and Y are Covalent Bonds, and Z is
--C.ident.C--
[0210] Similarly, following the procedure of 4A above, but
replacing
[9-((1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2--
yl)purin-6-yl]cyclopentylamine with
(1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxa
bicyclo[3.3.0]oct-2-yl)-purin-9-yl]oxolan-3-ylamine, the following
compounds of Formula I are prepared: [0211]
(4S,2R,3R,5R)-2-[6-(oxolan-3-ylamino)purin-9-yl]-5-ethynyloxolane-3,4-dio-
l; [0212]
(4S,2R,3R,5R)-2-[6-(cyclopentylmethylamino)purin-9-yl]-5-ethynyloxolane-3-
,4-diol; [0213]
(4S,2R,3R,5R)-2-[2-trifluoromethyl-6-(cyclopentylamino)purin-9-yl]-5-ethy-
nyloxolane-3,4-diol; [0214]
(4S,2R,3R,5R)-2-[6-(cyclobutylamino)purin-9-yl]-5-ethynyloxolane-3,4-diol-
; [0215]
(4S,2R,3R,5R)-2-[6-(cyclohexylamino)purin-9-yl]-5-ethynyloxolan-
e-3,4-diol; [0216]
(4S,2R,3R,5R)-2-[2-fluoro-6-(cyclohexylamino)purin-9-yl]-5-ethynyloxolane-
-3,4-diol; [0217]
(4S,2R,3R,5R)-2-[6-(cyclohexylmethylamino)purin-9-yl]-5-ethynyloxolane-3,-
4-diol; [0218]
(4S,2R,3R,5R)-2-[6-(3-fluorocyclopentylamino)purin-9-yl]-5-ethynyloxolane-
-3,4-diol; [0219]
(4S,2R,3R,5R)-2-[6-(4-trifluoromethylcyclopentylamino)purin-9-yl]-5-ethyn-
yloxolane-3,4-diol; [0220]
(4S,2R,3R,4R)-2-[6-(3-methoxycyclopentylamino)purin-9-yl]-5-ethynyloxolan-
e-3,4-diol; [0221]
(4S,2R,3R,5R)-2-[6-(phenylamino)purin-9-yl]-5-ethynyloxolane-3,4-diol;
[0222]
(4S,2R,3R,5R)-2-[6-(benzylamino)purin-9-yl]-5-ethynyloxolane-3,4--
diol; [0223]
(4S,2R,3R,5R)-2-[6-(4-fluorophenylamino)purin-9-yl]-5-ethynyloxolane-3,4--
diol; [0224]
(4S,2R,3R,5R)-2-[6-(pyridin-3-ylamino)purin-9-yl]-5-ethynyloxolane-3,4-di-
ol; [0225]
(4S,2R,3R,5R)-2-[6-(thiazol-2-ylamino)purin-9-yl]-5-ethynyloxolane-3,4-di-
ol; [0226]
(4S,2R,3R,5R)-2-[6-(tetrahydropyran-3-ylamino)purin-9-yl]-5-ethynyloxolan-
e-3,4-diol; [0227]
(4S,2R,3R,5R)-2-[6-(tetrahydropyran-3-ylmethylamino)purin-9-yl]-5-ethynyl-
oxolane-3,4-diol; and [0228]
(4S,2R,3R,5R)-2-[6-(5-fluorotetrahydropyran-3-ylamino)purin-9-yl]-5-ethyn-
yloxolane-3,4-diol.
C. Preparation of Compounds of Formula I, Varying X, R.sup.1 and
R.sup.2
[0229] Similarly, following the procedure of 4A above, but
replacing
[9-((1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2--
yl)purin-6-yl]cyclopentylamine with other compounds of formula (5),
other compounds of Formula I are prepared.
EXAMPLE 5
Preparation of a Compound of Formula (6)
A. Preparation of a Compound of Formula (6) where R.sup.1 is
Cyclopentyl, R.sup.2 is Hydrogen, R.sup.3 is
2-Trifluoromethylphenyl, Y is a Covalent Bond, and Z is
--C.ident.C--
[0230] ##STR19##
[0231] To a solution of
[9-((1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2--
yl)purin-6-yl]cyclopentylamine (40 mg, 0.12 mmol), a compound of
formula (5), in tetrahydrofuran (4 ml) under nitrogen was added
catalytic amounts (3 mg) of
dichlorobis(triphenylphosphine)palladium(II) and copper(II)iodide,
followed by 1-iodo-2-trifluoromethylbenzene (0.25 ml, 0.3 mmol).
Triethylamine (0.4 ml) was then added, and the mixture stirred for
15 minutes at room temperature. The solvent was removed under
reduced pressure, and the residue was purified by preparative TLC,
eluting with methanol:methylene chloride (6.5:1), to yield
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-(trifluoromethyl)-phenyl]ethynyl}--
3,6,8-trioxabicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine, a
compound of formula (6).
B. Preparation of a Compound of Formula (6) where R.sup.1 is
Cyclopentyl or Tetrahydrofuran-3-yl, R.sup.2, R.sup.4 and R.sup.5
are Hydrogen, X, X.sup.1 and Y are Covalent Bonds, and Z is
--C.ident.C--, Varying R.sup.3
[0232] Similarly, following the procedure of 5A above, but
replacing
[9-((1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct-2--
yl)purin-6-yl]cyclopentylamine with the appropriate compounds of
formula (5), the following compounds of formula (6) were prepared:
[0233]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-(trifluoromethyl)phenyl]ethynyl}-3-
,6,8-trioxabicyclo[3.3.0]oct-2-yl)purin-6-yl]oxolan-3-ylamine;
[0234]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl}-3,6,8-trioxa-
bicyclo[3.3.0]oct-2-yl)purin-6-yl]oxolan-3-ylamine; [0235]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-chlorophenyl]ethynyl}-3,6,8-trioxa-
bicyclo[3.3.0]oct-2-yl)purin-6-yl]oxolan-3-ylamine; [0236]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[thien-2-yl]ethynyl}-3,6,8-trioxabicy-
clo[3.3.0]oct-2-yl)purin-6-yl]oxolan-3-ylamine; [0237]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-chlorophenyl]ethynyl}-3,6,8-trioxa-
bicyclo[3.3.0]oct-2yl)purin-6-yl]cyclopentylamine; and [0238]
{9-[(1R,2R,4R,5R)-7,7-dimethyl-4-(2-(2-thienyl)ethynyl)-3,6,8-trioxabicyc-
lo[3.3.0]oct-2-yl]purin-6-yl}cyclopentylamine.
C. Preparation of a Compound of Formula (6), Varying R.sup.1,
R.sup.2, R.sup.3, R.sup.4, R.sup.5, X, X.sup.1 and Y, and Z is
--C.ident.C--
[0239] Similarly, following the procedure of 5A above, but
optionally replacing
[9-((1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.-
3.0]oct-2-yl)purin-6-yl]cyclopentylamine with other compounds of
formula (5), and optionally replacing
1-iodo-2-trifluoromethylbenzene with other compounds of formula
R.sup.3Y-LG, where LG is a leaving group, the following compounds
of Formula I are prepared: [0240]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-methylphenyl]ethynyl}-3,6,8-trioxa-
bicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine; [0241]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl}-3,6,8-trioxa-
bicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine; [0242]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[phenylethynyl]-3,6,8-trioxabicyclo[3-
.3.0]oct-2-yl)purin-6-yl]cyclopentylamine; [0243]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[5-chlorothien-2-yl]ethynyl}-3,6,8-tr-
ixoabicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine; [0244]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[4-methylisoxazol-3-yl]ethynyl}-3,6,9-
-trixoabicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine; [0245]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[3,5-dimethylisoxazol-4-yl]ethynyl}-3-
,6,8-trioxabicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine;
[0246]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[cyclopentyl]ethynyl}-3,6,8-trixoabic-
yclo[3.3.0]oct-2yl)purin-6-yl]cyclopentylamine; [0247]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorocyclohexyl]ethynyl}-3,6,8-tr-
ioxabicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine; [0248]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[piperidin-2-yl]ethynyl}-3,6,8-trixoa-
bicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine; [0249]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[4-methylpiperazin-1-yl]ethynyl}-3,6,-
8-trioxabicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine; [0250]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[pyridin-2-yl]ethynyl}-3,6,8-trixoabi-
cyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine; [0251]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[6-fluoropyridin-2-yl]ethynyl}-3,6,8--
trioxabicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine; [0252]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[thiazol-2-yl]ethynyl}-3,6,8-trixoabi-
cyclo[3.3.0]oct-2yl)purin-6-yl]cyclopentylamine; [0253]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[pyrimidin-2-yl]ethynyl}-3,6,8-trixoa-
bicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine; [0254]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-trixoab-
icyclo[3.3.0]oct-2-yl-purin-6-yl]cyclopentylmethylamine; [0255]
[9-((1R,2R,4R,5R)-2-trifluoromethyl-7,7-dimethyl-4-{2-[2-fluorophenyl]eth-
ynyl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl-purin-6-yl]cyclopentylmethylamine;
[0256]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,-
8-trixoabicyclo[3.3.0]oct-2-yl-purin-6-yl]cyclobutylamine; [0257]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-trixoab-
icyclo[3.3.0]oct-2yl-purin-6-yl]cyclohexylamine; [0258]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-trixoab-
icyclo[3.3.0]oct-2-yl-purin-6-yl](2-fluorocyclohexyl)amine; [0259]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-trixoab-
icyclo[3.3.0]oct-2-yl-purin-6-yl]cyclohexylmethylamine; [0260]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-trixoab-
icyclo[3.3.0]oct-2-yl-purin-6-yl](3-fluorocyclopentylamine; [0261]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-trixoab-
icyclo[3.3.0]oct-2-yl-purin-6-yl](4-trifluoromethylcyclopentylamine;
[0262]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-
-trixoabicyclo[3.3.0]oct-2-yl-purin-6-yl](3-methoxycyclopentylamine;
[0263]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-
-trixoabicyclo[3.3.0]oct-2-yl-purin-6-yl]phenylamine; [0264]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-trixoab-
icyclo[3.3.0]oct-2-yl-purin-6-yl](4-fluorophenyl)amine; [0265]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-trixoab-
icyclo[3.3.0]oct-2-yl-purin-6-yl]benzylamine; [0266]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-trixoab-
icyclo[3.3.0]oct-2-yl-purin-6-yl]pyridin-3-ylamine; [0267]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-fluorophenyl]ethynyl-3,6,8-trixoab-
icyclo[3.3.0]oct-2-yl-purin-6-yl]thiazol-2-ylamine; and [0268]
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[thien-2-yl]ethynyl}-3,6,8-trixoabicy-
clo[3.3.0]oct-2-yl)purin-6-yl](5-fluorooxolan-3-ylamine).
D. Preparation of Compounds of Formula I, Varying X, R.sup.1 and
R.sup.2
[0269] Similarly, following the procedure of 5A above, but
optionally replacing
[9-((1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.-
3.0]oct-2-yl)purin-6-yl]cyclopentylamine with other compounds of
formula (5), and optionally replacing
1-iodo-2-trifluoromethylbenzene with other compounds of formula
R.sup.3Y-LG, where LG is a leaving group, other compounds of
Formula I are prepared.
EXAMPLE 6
Preparation of a Compound of Formula I
A. Preparation of a Compound of Formula I where R.sup.1 is
Cyclopentyl, R.sup.2, R.sup.4 and R.sup.5 are Hydrogen, R.sup.3 is
2-Trifluoromethylphenyl, X, X.sup.1 and Y are Covalent Bonds, and Z
is --C.ident.C--
[0270] ##STR20##
[0271] a) To a solution of
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-ethynyloxolane-3,4-dio-
l (40 mg, 0.12 mmol) in tetrahydrofuran (4 mL) under nitrogen was
added catalytic amounts (3 mg) of
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-ethynyloxolane-3,4-dio-
l (II) and copper (I) iodide, followed by
1-iodo-2-trifluoromethylbenzene (0.042 mL). Triethylamine (0.4 mL)
was then added, and the mixture was stirred for 15 minutes at room
temperature. The solvent was removed under reduced pressure, and
the residue was purified by preparative TLC, eluting with
methanol:methylene chloride (6.5:1), to yield
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{2-[2-(trifluoromethyl
oxolane-3,4-diol, a compound of Formula I.
[0272] b) Alternatively, the acetonide protecting group was removed
from
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-(trifluoromethyl)-phenyl]ethynyl}--
3,6,8-trioxabicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine, a
compound of formula (6), in the same manner as shown in Example 4
to provide
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{2-[2-(trifluoromethyl-
)phenyl]ethynyl}oxolane-3,4-diol, a compound of Formula I.
B. Preparation of a Compound of Formula I where R.sup.1 is
Cyclopentyl or Tetrahydrofuran-3-yl, R.sup.2, R.sup.4 and R.sup.5
are Hydrogen, X, X.sup.1 and Y are Covalent Bonds, and Z is
--C.ident.C--, Varying R.sup.3
[0273] Similarly, following the procedure of 6A(a) above, but
optionally replacing
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-ethynyloxola-
ne-3,4-diol with other compounds of Formula I in which R.sup.3 is
hydrogen, and optionally replacing 1-iodo-2-trifluoromethylbenzene
with other compounds of formula R.sup.3Y-LG, where LG is a leaving
group, or:
[0274] Following the procedure of 6A(b) above, but replacing
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-(trifluoromethyl)-phenyl]ethynyl}
-3,6,8-trioxabicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine
with other compounds of formula (6); the following compounds of
Formula I were prepared: [0275]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{2-[2-fluorophenyl]-et-
hynyl}oxolane-3,4-diol; [0276]
(4S,2R,3R,5R)-2-[6-(oxalan-3-ylamino)purin-9-yl]-5-{2-[2-(trifluoromethyl-
)phenyl]-ethynyl}oxolane-3,4-diol; [0277]
(4S,2R,3R,5R)-2-[6-(oxalan-3-ylamino)purin-9-yl]-5-{2-[2-fluorophenyl]eth-
ynyl}oxolane-3,4-diol; [0278]
(4S,2R,3R,5R)-2-[6-(oxalan-3-ylamino)purin-9-yl]-5-{2-[2-chlorophenyl]eth-
ynyl}oxolane-3,4-diol; [0279]
(4S,2R,3R,5R)-2-[6-(oxalan-3-ylamino)purin-9-yl]-5-{2-[thien-2-yl]ethynyl-
}oxolane-3,4-diol; and [0280]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[thien-2-yl]ethynyl}-
oxolane-3,4-diol;
C. Preparation of a Compound of Formula I Varying R.sup.1 R.sup.2,
R.sup.3, R.sup.4, R.sup.4, X, X.sup.1 and Y, and Z is
--C.ident.C--
[0281] Similarly, following the procedure of 6A(a) above, but
optionally replacing
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-ethynyloxola-
ne-3,4-diol with other compounds of Formula I in which R.sup.3 is
hydrogen, and optionally replacing 1-iodo-2-trifluoromethylbenzene
with other compounds of formula R.sup.3Y-LG, where LG is a leaving
group, or:
[0282] Following the procedure of 6A(b) above, but replacing
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-(trifluoromethyl)-phenyl]ethynyl}--
3,6,8-trioxabicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine with
other compounds of formula (6); the following compounds of Formula
I are prepared: [0283]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[phenyl]ethynyl}oxol-
ane-3,4-diol; [0284]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[2-chlorophenyl]ethy-
nyl}oxolane-3,4-diol; [0285]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[2-methylphenyl]ethy-
nyl}oxolane-3,4-diol; [0286]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{5-chlorothien-2-yl]eth-
ynyl}oxolane-3,4-diol; [0287]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[4-methylisoxazol-3--
yl]ethynyl}oxolane-3,4-diol; [0288]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[2,5-dimethylisoxazo-
l-4-yl]ethynyl}oxolane-3,4-diol; [0289]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[cyclopentyl]ethynyl-
}oxolane-3,4-diol; [0290]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[2-fluorocyclohexyl]-
ethynyl}oxolane-3,4-diol; [0291]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[piperidin-2-yl]ethy-
nyl}oxolane-3,4-diol; [0292]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[4-methylpiperazin-1-
-yl]ethynyl}oxolane-3,4-diol; [0293]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[pyridin-2-yl]ethyny-
l}oxolane-3,4-diol; [0294]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[6-fluoropyridin-2-y-
l]ethynyl}oxolane-3,4-diol; [0295]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[thiazol-2-yl]ethyny-
l}-3,4-diol; [0296]
(4S,2R,3R,5R)-2-[6-cyclopentylamino)purin-9-yl]-5-{2-[pyrimidin-2-yl]ethy-
nyl}oxolane-3,4-diol; [0297]
(4S,2R,3R,5R)-2-[6-cyclopentylmethylamino)purin-9-yl]-5-{2-[2-fluoropheny-
l]ethynyl}oxolane-3,4-diol; [0298]
(4S,2R,3R,5R)-2-[6-(cyclopentylmethylamino)purin-9-yl]-5-{2-[2-fluorophen-
yl]-ethynyl}oxolane-3,4-diol; [0299]
(4S,2R,3R,5R)-2-[6-(cyclobutylamino)purin-9-yl]-5-{2-[2-fluorophenyl]-eth-
ynyl}oxolane-3,4-diol; [0300]
(4S,2R,3R,5R)-2-[6-(cyclohexylamino)purin-9-yl]-5-{2-[2-fluorophenyl]-eth-
ynyl}oxolane-3,4-diol; [0301]
(4S,2R,3R,5R)-2-[6-(2-fluorocyclohexylamino)purin-9-yl]-5-{2-[2-fluorophe-
nyl]-ethynyl}oxolane-3,4-diol; [0302]
(4S,2R,3R,5R)-2-[6-(cyclohexylmethylamino)purin-9-yl]-5-{2-[2-fluoropheny-
l]-ethynyl}oxolane-3,4-diol; [0303]
(4S,2R,3R,5R)-2-[6-(3-fluorocyclopentylamino)purin-9-yl]-5-{2-[2-fluoroph-
enyl]-ethynyl}oxolane-3,4-diol; [0304]
(4S,2R,3R,5R)-2-[6-(4-trifluoromethylcyclopentylamino)purin-9-yl]-5-}2-[2-
-fluorophenyl]-ethynyl}oxolane-3,4-diol; [0305]
(4S,2R,3R,5R)-2-[6-(3-methoxycyclopentylamino)purin-9-yl]-5-{2-[2-fluorop-
henyl]-ethynyl}oxolane-3,4-diol; [0306]
(4S,2R,3R,5R)-2-[6-(phenylamino)purin-9-yl]-5-{2-[2-fluorophenyl]-ethynyl-
}oxolane-3,4-diol; [0307]
(4S,2R,3R,5R)-2-[6-(4-fluorophenylamino)purin-9-yl]-5-{2-[2-fluorophenyl]-
-ethynyl}oxolane-3,4-diol; [0308]
(4S,2R,3R,5R)-2-[6-(benzylamino)purin-9-yl]-5-{2-[2-fluorophenyl]-ethynyl-
}oxolane-3,4-diol; [0309]
(4S,2R,3R,5R)-2-[6-(pyridin-3-ylamino)purin-9-yl]-5-{2-[2-fluorophenyl]-e-
thynyl}oxolane-3,4-diol; [0310]
(4S,2R,3R,5R)-2-[6-(thiazol-2-ylamino)purin-9-yl]-5-{2-[2-fluorophenyl]et-
hynyl}oxolane-3,4-diol; [0311]
(4S,2R,3R,5R)-2-[6-(oxolan-3-ylmethylamino)purin-9-yl]-5-{2-[2-fluorophen-
yl]-ethynyl}oxolane-3,4-diol; and [0312]
(4S,2R,3R,5R)-2-[6-(5-fluorooxolan-3-ylamino)purin-9-yl]-5-{2-[2-fluoroph-
enyl]-ethynyl}oxolane-3,4-diol.
D. Preparation of a Compound of Formula I Varying R.sup.1 R.sup.2,
R.sup.3, R.sup.4, R.sup.5, X, X.sup.1 and Y, and Z is
--C.ident.C--
[0313] Similarly, following the procedure of 6A above, but
replacing
[9-((1R,2R,4R,5R)-7,7-dimethyl-4-{2-[2-(trifluoromethyl)-phenyl]ethynyl}--
3,6,8-trioxabicyclo[3.3.0]oct-2-yl)purin-6-yl]cyclopentylamine with
other compounds of formula (6), other compounds of Formula I are
prepared.
EXAMPLE 7
Preparation of a Compound of Formula (7)
Preparation of a Compound of Formula (7) where R.sup.1 is
Tetrahydrofuran-3-yl, R.sup.2 is Hydrogen, R.sup.3 is
4-Fluorophenyl, X and Y are Covalent Bonds, and Z is
--CH.dbd.CH--
[0314] ##STR21##
[0315] To a solution of
(2S,1R,4R,5R)-7,7-dimethyl-3,6,8-trioxa-4-[6-(oxolan-3-ylamino)purin-9-yl-
]bicyclo[3.3.0]octane-2-carbaldehyde, a compound of formula (4)
(200 mg), in methylene chloride (5 ml) was added
(4-fluorophenyl)triphenylphosphonium bromide (459 mg, 1 mmol),
followed by dropwise addition of an aqueous solution of 50% sodium
hydroxide. After addition was complete, the mixture was stirred for
2 hours, then washed with water. The organic layer was separated,
dried, and solvent removed under reduced pressure. The residue was
purified by preparative thin layer chromatography, eluting with
ethyl acetate, to give pure
(9-{4-[2-(4-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)oxolan-3-ylamine, a compound of
formula (7).
B. Preparation of a Compound of Formula (7) where R.sup.1 is
Cyclopentyl or Tetrahydrofuran-3-yl, R.sup.2, R.sup.4 and R.sup.5
are Hydrogen, X, X.sup.1 and Y are Covalent Bonds, and Z is
--CH.dbd.CH--, Varying R.sup.3
[0316] Similarly, following the procedure of 7A above, but
optionally replacing
(2S,1R,4R,5R)-7,7-dimethyl-3,6,8-trioxa-4-[6-(oxolan-3-ylamino)-
purin-9-yl]bicyclo[3.3.0]octane-2-carbaldehyde with other compounds
of formula (4), and optionally replacing
(4-fluorophenyl)triphenylphosphonium bromide with other compounds
of formula R.sup.3YCH.sub.2P(PH).sub.3Br, the following compounds
of formula (7) were prepared: [0317]
(9-{4-[2-(5-chlorothien-2-yl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-triox-
abicyclo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0318]
(9-{4-[2-(3,5-dimethylisoxazol-4-yl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-
-trioxabicyclo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0319]
(9-{4-[2-(4-methylisoxazol-3-yl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-tri-
oxabicyclo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0320]
(9-{4-[2-(2-methylphenyl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicy-
clo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; and [0321]
(9-{4-[2-(phenyl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.-
0]oct-2-yl}purin-6-yl)cyclopentylamine.
C. Preparation of a Compound of Formula (7) Varying R.sup.1,
R.sup.2, R.sup.3, R.sup.4, R.sup.5, X, X.sup.1, Y, and Z is
--CH.dbd.CH--
[0322] Similarly, following the procedure of 7A above, but
optionally replacing
(2S,1R,4R,5R)-7,7-dimethyl-3,6,8-trioxa-4-[6-(oxolan-3-ylamino)-
purin-9-yl]bicyclo[3.3.0]octane-2-carbaldehyde with other compounds
of formula (4), and optionally replacing
(4-fluorophenyl)triphenylphosphonium bromide with other compounds
of formula R.sup.3YCH.sub.2P(PH).sub.3Br, the following compounds
of Formula I are prepared: [0323]
(9-{4-[2-(2-methylphenyl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicy-
clo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0324]
(9-{4-[2-(2-fluorophenyl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicy-
clo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0325]
(9-{4-[2-(phenyl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.-
0oct-2-yl}purin-6-yl)cyclopentylamine; [0326]
(9-{4-[2-(cyclopentyl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicyclo-
[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0327]
(9-{4-[2-(2-fluorocyclohexyl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxa-
bicyclo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0328]
(9-{4-[2-(piperidin-2-yl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicy-
clo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0329]
(9-{4-[2-(4-methylpiperazin-1-yl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-tr-
ioxabicyclo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0330]
(9-{4-[2-(pyridin-2-yl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicycl-
o[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0331]
(9-{4-[2-(6-fluoropyridin-2-yl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0332]
(9-{4-[2-(thiazol-2-yl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicycl-
o[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0333]
(9-{4-[2-(pyrimidin-2-yl]vinyl(1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicy-
clo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine; [0334]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylmethylamine; [0335]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-2-trifluorometh-
yl-3,6,8-trioxabicyclo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylmethylamine;
[0336]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-t-
rioxabicyclo[3.3.0]oct-2-yl}purin-6-yl)cyclobutylmethylamine;
[0337]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)cyclohexylamine; [0338]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)(2-fluorocyclohexylamine); [0339]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)cyclohexylmethylamine; [0340]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)(3-fluorocyclopentylamine); [0341]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)(4-trifluoromethylcyclopentylamine);
[0342]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-t-
rioxabicyclo[3.3.0]oct-2-yl}purin-6-yl)(3-methoxycyclopentylamine);
[0343]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-t-
rioxabicyclo[3.3.0]oct-2-yl}purin-6-yl)phenylamine; [0344]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)(4-fluorophenylamine); [0345]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)benzylamine; [0346]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)pyridin-3-ylamine); [0347]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)thiazol-2-ylamine; and [0348]
(9-{4-[2-(2-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabic-
yclo[3.3.0]oct-2-yl}purin-6-yl)(5-fluorooxolan-3-ylamine).
D. Preparation of a Compound of Formula (7) Varying R.sup.1,
R.sup.2, R.sup.3, R.sup.4, R.sup.5, X, X.sup.1, Y, and Z is
--CH.dbd.CH--
[0349] Similarly, following the procedure of 7A above, but
optionally replacing
(2S,1R,4R,5R)-7,7-dimethyl-3,6,8-trioxa-4-[6-(oxolan-3-ylamino)-
purin-9-yl]bicyclo[3.3.0]octane-2-carbaldehyde with other compounds
of formula (4), and optionally replacing
(4-fluorophenyl)triphenylphosphonium bromide with other compounds
of formula R.sup.3YCH.sub.2P(PH).sub.3Br, other compounds of
Formula I are prepared.
EXAMPLE 8
Preparation of a Compound of Formula I
A. Preparation of a Compound of Formula I where R.sup.1 is
Tetrahydrofuran-3-yl, R.sup.2 is Hydrogen, R.sup.3 is
4-Fluorophenyl, X and Y are Covalent Bonds, and Z is
--CH.dbd.CH--
[0350] ##STR22##
[0351] The acetonide protecting group was then removed from
(9-{4-[(1E)-2-(4-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}purin-6-yl)oxolan-3-ylamine, a compound of
formula (7), in the same manner as shown in Example 4 to provide
5-[2-(4-fluorophenyl)vinyl]-2-[6-(oxolan-3-ylamino)purin-9-yl]oxolane-3,4-
-diol, a compound of Formula I.
B. Preparation of a Compound of Formula I where R.sup.1 is
Cyclopentyl or Tetrahydrofuran-3-yl, R.sup.2, R.sup.4 and R.sup.5
are Hydrogen, X, X.sup.1 and Y are Covalent Bonds, and Z is
--CH.dbd.CH--, Varying R.sup.3
[0352] Similarly, following the procedure of 8A above, but
replacing
(9-{4-[(1E)-2-(4-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}purin-6-yl)oxolan-3-ylamine with other
compounds of formula (7), the following compounds of Formula I were
prepared: [0353]
5-[(1E)-2-(methoxycarbonylvinyl](4S,2R,3R,5R)-2-[6-(oxolan-3-ylam-
ino)purin-9-yl]oxolane-3,4-diol; [0354]
5-[(1E)-2-(2-methylphenyl)vinyl](4S,2R,3R,5R)-2-[6-(oxolan-3-ylamino)puri-
n-9-yl]oxolane-3,4-diol; [0355]
5-[2-(5-chlorothien-2-yl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-
-9-yl]oxolane-3,4-diol; [0356]
5-[2-(3,5-dimethylisoxazol-4-yl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamin-
o)purin-9-yl]oxolane-3,4-diol; [0357]
5-[2-(4-methylisoxazol-3-yl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)pu-
rin-9-yl]oxolane-3,4-diol; [0358]
5-[2-(2-methylphenyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-y-
l]oxolane-3,4-diol; and [0359]
5-[2-(phenyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]oxolan-
e-3,4-diol;
C. Preparation of a Compound of Formula I Varying R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, X, X.sub.1, Y, and Z is
--CH.dbd.CH--
[0360] Similarly, following the procedure of 8A above, but
replacing
(9-{4-[(1E)-2-(4-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}purin-6-yl)oxolan-3-ylamine with other
compounds of formula (7), the following compounds of Formula I are
prepared: [0361]
5-[2-(2-methylphenyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)p-
urin-9-yl]oxolane-3,4-diol, [0362]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-y-
l]oxolane-3,4-diol, [0363]
5-[2-(phenyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]oxolan-
e-3,4-diol, [0364]
5-[2-(cyclopentyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]o-
xolane-3,4-diol, [0365]
5-[2-(2-fluorocyclohexyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-
-9-yl]oxolane-3,4-diol, [0366]
5-[2-(2-piperidin-2-yl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-
-yl]oxolane-3,4-diol, [0367]
5-[2-(4-methylpiperazin-1-yl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)p-
urin-9-yl]oxolane-3,4-diol, [0368]
5-[2-(2-pyridin-2-yl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-y-
l]oxolane-3,4-diol, [0369]
5-[2-(6-fluoropyridin-2-yl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)pur-
in-9-yl]3,4-diol, [0370]
5-[2-(2-thiazol-2-yl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-y-
l]oxolane-3,4-diol, [0371]
5-[2-(pyrimidin-2-yl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-y-
l]oxolane-3,4-diol, [0372]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclopentylmethylamino)pur-
in-9-yl]oxolane-3,4-diol, [0373]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclobutylmethylamino)puri-
n-9-yl]oxolane-3,4-diol, [0374]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(cyclohexylamino)purin-9-yl-
]oxolane-3,4-diol, [0375]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(2-fluorocyclohexylamino)pu-
rin-9-yl]oxolane-3,4-diol, [0376]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(2-fluorocyclohexylmethylam-
ino)purin-9-yl]oxolane-3,4-diol, [0377]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(3-fluorocyclopentylamino)p-
urin-9-yl]oxolane-3,4-diol, [0378]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(4-trifluoromethylcyclopent-
ylamino)purin-9-yl]oxolane-3,4-diol, [0379]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(3-methoxycyclopentylamino)-
purin-9-yl]oxolane-3,4-diol, [0380]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(phenylamino)purin-9-yl]oxo-
lane-3,4-diol, [0381]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(benzylamino)purin-9-yl]oxo-
lane-3,4-diol, [0382]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(pyridin-3-ylamino)purin-9--
yl]oxolane-3,4-diol, [0383]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(thiazol-2-ylamino)purin-9--
yl]oxolane-3,4-diol, and [0384]
5-[2-(2-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(5-fluorooxolan-3-ylamino)p-
urin-9-yl]oxolane-3,4-diol.
D. Preparation of a Compound of Formula I Varying R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, X, X.sup.1, Y, and Z is
--CH.dbd.CH--
[0385] Similarly, following the procedure of 8A above, but
replacing
(9-{4-[(1E)-2-(4-fluorophenyl)vinyl](1R,2R,4R,5R)-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}purin-6yl)oxolan-3-ylamine with other
compounds of formula (7), other compounds of Formula I are
prepared.
EXAMPLE 9
Preparation of a Compound of Formula I
Preparation of a Compound of Formula I where R.sup.1 is
Tetrahydrofuran-3-yl, R.sup.2 is Hydrogen, R.sup.3 is
4-Fluorophenyl, X and Y are Covalent Bonds, and Z is
--CH.sub.2CH.sub.2--
[0386] ##STR23##
[0387] To a solution of
5-[2-(4-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(oxolan-3-ylamino)purin-9-y-
l]oxolane-3,4-diol (40 mg) in ethanol (5 ml) and cyclohexene (2 ml)
was added palladium hydroxide (50 mg), and the mixture was stirred
for 24 hours. The catalyst was filtered off, and solvent removed
under reduced pressure. The residue was purified by preparative
thin layer chromatography, to give pure
(4S,2R,3R,5R)-5-[2-(4-fluorophenyl)ethyl]-2-[6-(oxolan-3-ylamino)purin-9--
yl]oxolane-3,4-diol, a compound of Formula I.
B. Preparation of a Compound of Formula I where R.sup.1 is
Cyclopentyl or Tetrahydrofuran-3-yl, R.sup.2, R.sup.4 and R.sup.5
are Hydrogen, X, X.sup.1 and Y are Covalent Bonds, and Z is
--CH.sub.2CH.sub.2--, Varying R.sup.3
[0388] Similarly, following the procedure of 9A above, but
replacing
5-[2-(4-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(oxolan-3-ylamino)purin-9-y-
l]oxolane-3,4-diol with other compounds of Formula I in which Z is
--CH.dbd.CH--, the following compounds of Formula I were prepared:
[0389]
(4S,2R,3R,5R)-5-[2-(methoxycarbonyl)ethyl]-2-[6-(oxolan-3-ylamino-
)purin-9-yl]oxolane-3,4-diol; [0390]
(4S,2R,3R,5R)-5-[2-(2-methylphenyl)ethyl]2-[6-(oxolan-3-ylamino)purin-9-y-
l]oxolane-3,4-diol; [0391]
(4S,2R,3R,5R)-5-[2-phenylethyl]2-[6-(cyclopentylamino)purin-9-yl]oxolane--
3,4-diol; [0392]
(4S,2R,3R,5R)-5-[2-(3,5-dimethylisoxazol-4-yl)ethyl]2-[6-(cyclopentylylam-
ino)purin-9-yl]oxolane-3,4-diol; and [0393]
(4S,2R,3R,5R)-5-[2-(5-chlorothien-2-yl)ethyl]
2-[6-(cyclopentylamino)purin-9-yl]oxolane-3,4-diol.
C. Preparation of a Compound of Formula I Varying R.sup.1 R.sup.2,
R.sup.3, R.sup.4 and R.sup.5 X, X.sup.1, Y, and Z is
--CH.sub.2CH.sub.2--
[0394] Similarly, following the procedure of 9A above, but
replacing
5-[2-(4-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(oxolan-3-ylamino)purin-9-y-
l]oxolane-3,4-diol with other compounds of Formula I in which Z is
--CH.dbd.CH--, the following compounds of Formula I are prepared:
[0395]
(4S,2R,3R,5R)-5-[2-(4-methylisoxazol-3-yl)ethyl]-2-[6-(cyclopentylamino)-
purin-9-yl]oxolane-3,4-diol; [0396]
(4S,2R,3R,5R)-5-[2-(2-methylphenyl)ethyl]-2-[6-(cyclopentylylamino)purin--
9-yl]oxolane-3,4-diol; [0397]
(4S,2R,3R,5R)-5-[2-(2-methylphenyl)ethyl]-2-[6-(oxolan-3-ylamino)purin-9--
yl]oxolane-3,4-diol; [0398]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(cyclopentylamino)purin-9-y-
l]oxolane-3,4-diol, [0399]
(4S,2R,3R,5R)-5[2-(phenyl)ethyl]-2-[6-(cyclopentylamino)purin-9-yl]oxolan-
e-3,4-diol, [0400]
(4S,2R,3R,5R)-5[2-(cyclopentyl)ethyl]-2-[6-(cyclopentylamino)purin-9-yl]o-
xolane-3,4-diol, [0401]
(4S,2R,3R,5R)-5[2-(2-fluorocyclohexyl)ethyl]-2-[6-(cyclopentylamino)purin-
-9-yl]oxolane-3,4-diol, [0402]
(4S,2R,3R,5R)-5[2-(2-piperidin-2-yl)ethyl]-2-[6-(cyclopentylamino)purin-9-
-yl]oxolane-3,4-diol, [0403]
(4S,2R,3R,5R)-5[2-(4-methylpiperazin-1-yl)ethyl]-2-[6-(cyclopentylamino)p-
urin-9-yl]oxolane-3,4-diol, [0404]
(4S,2R,3R,5R)-5[2-(2-pyridin-2-yl)ethyl]-2-[6-(cyclopentylamino)purin-9-y-
l]oxolane-3,4-diol, [0405]
(4S,2R,3R,5R)-5[2-(6-fluoropyridin-2-yl)ethyl]-2-[6-(cyclopentylamino)pur-
in-9-yl]oxolane-3,4-diol, [0406]
(4S,2R,3R,5R)-5[2-(2-thiazol-2-yl)ethyl]-2-[6-(cyclopentylamino)purin-9-y-
l]oxolane-3,4-diol, [0407]
(4S,2R,3R,5R)-5[2-(pyrimidin-2-yl)ethyl]-2-[6-(cyclopentylamino)purin-9-y-
l]oxolane-3,4-diol, [0408]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-.sup.2-[6-(cyclopentylmethylamin-
o)purin-9-yl]oxolane-3,4-diol, [0409]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(cyclobutylmethylamino)puri-
n9-yl]oxolane-3,4-diol, [0410]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(cyclohexylamino)purin-9-yl-
]oxolane-3,4-diol, [0411] (4S,2R,3R,5R)-5[2-(2-fluorophenyl)
ethyl]-2-[6-(2-fluorocyclohexylamino)purin-9-yl]oxolane-3,4-diol,
[0412]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(2-fluorocyclohexylmethyla-
mino)purin-9-yl]oxolane-3,4-diol, [0413]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(3-fluorocyclopentylamino)p-
urin-9-yl]oxolane-3,4-diol, [0414]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(4-trifluoromethylcyclopent-
ylamino)purin-9-yl]oxolane-3,4-diol, [0415]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(3-methoxycyclopentylamino)-
purin-9-yl]oxolane-3,4-diol, [0416]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(phenylamino)purin-9-yl]oxo-
lane-3,4-diol, [0417]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(benzylamino)purin-9-yl]oxo-
lane-3,4-diol, [0418]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(pyridin-3-ylamino)purin-9--
yl]oxolane-3,4-diol, [0419]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(thiazol-2-ylamino)purin-9--
yl]oxolane-3,4-diol, and [0420]
(4S,2R,3R,5R)-5[2-(2-fluorophenyl)ethyl]-2-[6-(5-fluorooxolan-3-ylamino)p-
urin-9-yl]oxolane-3,4-diol.
D. Preparation of a Compound of Formula I Varying R.sup.1 R.sup.2,
R.sup.3, R.sup.4 and R.sup.5, X, X.sup.1, Y, and Z is
--CH.sub.2CH.sub.2--
[0421] Similarly, following the procedure of 9A above, but
replacing
5-[2-(4-fluorophenyl)vinyl](4S,2R,3R,5R)-2-[6-(oxolan-3-ylamino)purin-9-y-
l]oxolane-3,4-diol with other compounds of Formula I in which Z is
--CH.dbd.CH--, other compounds of Formula I are prepared.
EXAMPLE 10
Preparation of a Compound of Formula (8) where R.sup.1 is
Cyclopentyl and R.sup.2 is Hydrogen
[0422] ##STR24##
[0423] To a stirred solution of benzyloxymethyltriphenyl
phosphonium chloride (7.71 g, 18.40 mmol) in tetrahydrofuran (40
mL) was added N-sodiumhexamethyldisilazane (1M in THF, 17.50 mL)
dropwise, and the mixture was stirred at -78.degree. C. for 1 hour.
Then
{(2S,1R,4R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trioxab-
icyclo[3.3.0]oct-2-yl}formaldehyde, a compound of formula (4) (3.73
g, 10 mmol), was dissolved in tetrahydrofuran (10 mL) and added
slowly. The mixture was slowly brought to room temperature, and
stirred overnight, quenched with H.sub.2O (60 mL) and extracted
with ethyl acetate (3.times.50 mL). The combined organic layers was
washed with water (2.times.50 mL), dried over Na.sub.2SO.sub.4, and
evaporated under reduced pressure. Column chromatography gave
(9-{4-[2-(phenylmethoxy)vinyl](2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicyclo-
[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine, a compound of formula
(8).
EXAMPLE 11
Preparation of a Compound of Formula (9) where R.sup.1 is
Cyclopentyl and R.sup.2 is Hydrogen
[0424] ##STR25##
[0425] A mixture of
(9-{4-[(1E)-2-(phenylmethoxy)vinyl](2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabi-
cyclo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine, a compound of
formula (8)(1.90 g, 3.98 mmol) and catalytic amount of Pd/C in
methanol (30 mL) was stirred under hydrogen (67 psi) overnight. The
Pd/C was filtered through celite and washed with methanol. The
filtrate was concentrated and the residue,
(9-{4-[(1E)-2-(phenylmethoxy)vinyl](2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabi-
cyclo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine, a compound of
formula (9), was used without further purification.
EXAMPLE 12
Preparation of a Compound of Formula (10) where R.sup.1 is
Cyclopentyl and R.sup.2 is Hydrogen
[0426] ##STR26##
[0427] A mixture of
(9-{4-[(1E)-2-(phenylmethoxy)vinyl](2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabi-
cyclo[3.3.0]oct-2-yl}purin-6-yl)cyclopentylamine, a compound of
formula (9), and Pd(OH).sub.2 in cyclohexene (45 mL) and ethanol
(30 mL) was stirred at 80.degree. C. for 5 days. The mixture was
filtered through celite and washed with ethanol. The filtrate was
concentrated under reduced pressure, and the residue purified by
column chromatography to give
2-{(1R,2R,4R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8--
trioxabicyclo[3.3.0]oct-2-yl}ethan-1-ol, a compound of the Formula
(10).
EXAMPLE 13
Preparation of a Compound of Formula (11) where R.sup.1 is
Cyclopentyl and R.sup.2 is Hydrogen
[0428] ##STR27##
[0429] To an ice-cooled solution of
2-{((1R,2R,4R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-trio-
xabicyclo[3.3.0]oct-2-yl}ethan-1-ol, a compound of the Formula (10)
(0.90 g, 2.31 mmol), dicyclohexylcarbodiimide (1.43 g, 6.93 mmol)
and pyridine (0.19 mL, 2.31 mmol) in dimethylsulfoxide (16 mL) was
added trifluoroacetic acid (0.09 mL, 1.15 mmol) slowly. The
resulting mixture was stirred under N.sub.2 at room temperature
overnight. N,N'-dicyclohexylurea was then filtered off, and washed
with ethyl acetate (60 mL). The filtrate was washed with water
(3.times.40 mL), dried over Na.sub.2SO.sub.4, and concentrated
under reduced pressure. The residue,
2-{(1R,2R,4R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,-
6,8-trioxabicyclo[3.3.0]oct-2-yl}ethan-1-one, a compound of Formula
(11) was used without further purification.
EXAMPLE 14
Preparation of a Compound of Formula (12) where R.sup.1 is
Cyclopentyl and R.sup.2 is Hydrogen
[0430] ##STR28##
[0431] To a stirred solution of potassium-tert-butoxide (0.78 g,
6.93 mmol) in tetrahydrofuran (45 mL) was added
(bromomethyl)triphenylphosphonium bromide (1.51 g, 3.27 mmol) in
small portions at -78.degree. C. The resulting mixture was stirred
at -78.degree. C. for 2 hours.
2-{(1R,2R,4R)-4-[6-(cyclopentylamino)purin-9-yl]-7,7-dimethyl-3,6,8-triox-
abicyclo[3.3.0]oct-2-yl} ethan-1-one, a compound of Formula (11)
(0.89 g, 2.31 mmol) dissolved in THF (15 mL) was added slowly. and
the mixture stirred under an atmosphere of N.sub.2 at -78.degree.
C. for 3 hours, brought to room temperature slowly and stirred for
4 days. The reaction mixture was quenched with saturated NH.sub.4Cl
aqueous solution (15 mL) and diluted with H.sub.2O (60 mL), and
then extracted with EtOAc (3.times.50 mL). The combined organic
layers were washed with H.sub.2O (2.times.50 mL), dried over
Na.sub.2SO.sub.4, and evaporated under reduced pressure, to provide
{9-[4-((2E)-3-bromoprop-2-enyl)(2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicycl-
o[3.3.0]oct-2-yl]purin-6-yl}cyclopentylamine, a compound of formula
(12), which was isolated by column chromatography.
[0432] To a stirred solution of
{9-[4-((2E)-3-bromoprop-2-enyl)(2R,4R,5R)-7,7-dimethyl-3,6,8-trioxabicycl-
o[3.3.0]oct-2-yl]purin-6-yl}cyclopentylamine, (0.40 g, 0.86 mmol)
in tetrahydrofuran (25 mL) was added potassium-tert-butoxide (0.78
g, 6.93 mmol) in small portions at -78.degree. C. The resulting
mixture was stirred at -78.degree. C. for 30 minutes, then brought
to room temperature slowly and stirred overnight. The reaction
mixture was quenched with saturated aqueous ammonium chloride
solution (5 mL) and diluted with H.sub.2O (30 mL), then extracted
with EtOAc (3.times.30 mL). The combined organic layers were washed
with water (2.times.30 mL), dried over sodium sulfate, filtered,
and the filtrate evaporated under reduced pressure, to provide
[9-((2R,4R,5R)-7,7-dimethyl-3,6,8-trioxa-4-prop-2-ynylbicyclo[3.3.0]oct-2-
-yl)purin-6-yl]cyclopentylamine, a compound of formula (13), which
was isolated by column chromatography.
EXAMPLE 15
Preparation of a Compound of Formula I
Preparation of a Compound of Formula I where R.sup.1 is
Cyclopentyl, R.sup.2 and R.sup.3 are Hydrogen, X and Y are Covalent
Bonds, X.sup.1 is --CH.sub.2-- and Z is --C.ident.C--
[0433] ##STR29##
[0434]
[9-((2R,4R,5R)-7,7-dimethyl-3,6,8-trioxa-4-prop-2-ynylbicyclo[3.3.-
0]oct-2-yl)purin-6-yl]cyclopentylamine, a compound of formula (13),
was stirred in 80% acetic acid aqueous solution (20 mL) at
80.degree. C. in a sealed tube for 2 days. The solvent was removed
under reduced pressure, and the residue purified by preparative
thin layer chromatography, to give
(4S,2R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-prop-2-ynyloxolane-3-
,4-diol, a compound of Formula I.
EXAMPLE 16
Preparation of a Compound of Formula I
A. Preparation of a Compound of Formula I where R.sup.1 is
Cyclopentyl, R.sup.2 is Hydrogen. R.sup.3 is 2-Fluorophenyl, X and
Y are Covalent Bonds, X.sup.1 is --CH.sub.2-- and Z is
--C.ident.C--
[0435] ##STR30##
[0436] To a stirred solution of
(4S,2R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-prop-2-ynyloxolane-3,4-di-
ol (0.017 g, 0.05 mmol) and 1-fluoro-2-iodobenzene (0.02 mL, 0.13
mmol) in THF (3.50 mL) under an atmosphere of N.sub.2 was added
catalytic amount of dichlorobis(triphenylphosphine)palladium (II)
and copper (I) iodide. Et.sub.3N (0.20 mL) was then added. The
resulting mixture was stirred under N.sub.2 at 75.degree. C. for 2
days. The solvent was removed under reduced pressure, and the
residue was purified by preparative thin layer chromatography to
give
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-[3-(2-fluorophenyl)pro-
p-2-ynyl]oxolane-3,4-diol, a compound of Formula I.
B. Preparation of a Compound of Formula I where R.sup.1 is
Cyclopentyl, R.sup.2 R.sup.4 and R.sup.5 are Hydrogen, X and
X.sup.1 are Covalent Bonds, Y is CH.sup.2, and Z is --C.ident.C--,
varying R.sup.3
[0437] Similarly, following the procedures of Examples 10-16 above,
the following compounds of Formula I in which X.sup.1 is
--CH.sub.2-- and Z is --C.ident.C-- were prepared: [0438]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[2-fluorophenyl]pro-
p-2-ynyl}oxolane-3,4-diol; and [0439]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[2-(trifluoromethyl-
)phenyl]prop-2-ynyl}oxolane-3,4-diol.
C. Preparation of a Compound of Formula I Varying R.sup.1, R.sup.2,
R.sup.3 R.sup.4 and R.sup.5, X, X.sup.1 and Y is CH.sub.2, and Z is
--C.ident.C--
[0440] Similarly, following the procedures of Examples 10-16 above,
the following compounds of Formula I in which X.sup.1 is
--CH.sub.2-- and Z is --C.ident.C-- are prepared: [0441]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[4-methylisoxazol-3-
-yl]prop-2-ynyl}oxolane-3,4-diol; [0442]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[2-methylphenyl]pro-
p-2-ynyl}oxolane-3,4-diol; [0443]
(4S,2R,3R,5R)-2-[6-(oxolan-3-ylamino)purin-9-yl]-5-{3-[2-fluorophenyl]pro-
p-2-ynyl}oxolane-3,4-diol; [0444]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[2-fluorophenyl]pro-
p-2-ynyl}oxolane-3,4-diol; [0445]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[phenyl]prop-2-ynyl-
}oxolane-3,4-diol; [0446]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[cyclopentyl]prop-2-
-ynyl}oxolane-3,4-diol; [0447]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[2-fluorocyclohexyl-
]prop-2-ynyl}oxolane-3,4-diol; [0448]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[piperidin-2-yl]pro-
p-2-ynyl}oxolane-3,4-diol; [0449]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[4-methylpiperazin--
1yl]prop-2-ynyl}oxolane-3,4-diol; [0450]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[pyridin-2-yl]prop--
2-ynyl}oxolane-3,4-diol; [0451]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[6-fluoropyridin-2--
yl]prop-2-ynyl}oxolane-3,4-diol; [0452]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[thiazol-2-yl]prop--
2-ynyl}oxolane-3,4-diol; [0453]
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{3-[pyrimidin-2-yl]pro-
p-2-ynyl}oxolane-3,4-diol; [0454]
(4S,2R,3R,5R)-2-[6-(cyclopentylmethylamino)purin-9-yl]-5-{3-[2-fluorophen-
yl]prop-2-ynyl}oxolane-3,4-diol; [0455]
(4S,2R,3R,5R)-2-[6-(cyclobutylmethylamino)purin-9-yl]-5-{3-[2-fluoropheny-
l]prop-2-ynyl}oxolane-3,4-diol; [0456]
(4S,2R,3R,5R)-2-[6-(cyclohexylamino)purin-9-yl]-5-{3-[2-fluorophenyl]prop-
-2-ynyl}oxolane-3,4-diol; [0457]
(4S,2R,3R,5R)-2-[6-(2-fluorocyclohexylamino)purin-9-yl]-5-{3-[2-fluorophe-
nyl]prop-2-ynyl}oxolane-3,4-diol; [0458]
(4S,2R,3R,5R)-2-[6-(2-fluorocyclohexylmethylamino)purin-9-yl]-5-{3-[2fluo-
rophenyl]prop-2-ynyl}oxolane-3,4-diol; [0459]
(4S,2R,3R,5R)-2-[6-(3-fluorocyclopentylamino)purin-9-yl]-5-{3-[2-fluoroph-
enyl]prop-2-ynyl}oxolane-3,4-diol; [0460]
(4S,2R,3R,5R)-2-[6-(4-trifluoromethylcyclopentylamino)purin-9-yl]-5-{3-[2-
-fluorophenyl]prop-2-ynyl}oxolane-3,4-diol; [0461]
(4S,2R,3R,5R)-2-[6-(phenylamino)purin-9-yl]-5-{3-[2-fluorophenyl]prop-2-y-
nyl}oxolane-3,4-diol; [0462]
(4S,2R,3R,5R)-2-[6-(benzylamino)purin-9-yl]-5-{3-[2-fluorophenyl]prop-2-y-
nyl}oxolane-3,4-diol; [0463]
(4S,2R,3R,5R)-2-[6-(pyridin-3-ylamino)purin-9-yl]-5-{3-[2-fluorophenyl]pr-
op-2-ynyl}oxolane-3,4-diol; [0464]
(4S,2R,3R,5R)-2-[6-(thiazol-2-ylamino)purin-9-yl]-5-{3-[2-fluorophenyl]pr-
op-2ynyl}oxolane-3,4-diol; and [0465]
(4S,2R,3R,5R)-2-[6-(5-fluorooxolan-3-ylamino)purin-9-yl]-5-{3-[2-fluoroph-
enyl]prop-2-ynyl}oxolane-3,4-diol.
D. Preparation of a Compound of Formula I Varying R.sup.1, R.sup.2,
R.sup.2, R.sup.4 and R.sup.5, X, X.sup.1 and Y is CH.sub.2, and Z
is --C.ident.C--
[0466] Similarly, following the procedures of Examples 10-16 above,
other compounds of Formula I in which X.sup.1 is --CH.sub.2-- and Z
is --C.ident.C-- are prepared.
EXAMPLE 17
Preparation of a Compound of Formula I
A. Preparation of a Compound of Formula I where R.sup.1 is
2-Hydroxycyclopentyl, R.sup.2 is Hydrogen, R.sup.3 is Hydrogen, X,
X.sup.1 and Y are Covalent Bonds, and Z is --C.ident.C--
[0467] ##STR31##
[0468] 1) To a suspension of potassium t-butoxide (0.84 g, 7.5
mmol) in tetrahydrofuran (5 ml) at -78.degree. C. was added
bromomethyltriphenylphosphonium (1.64 g, 3.75 mmol) in small
portions, and the mixture stirred for 2 hours. To this mixture was
added a solution of
(2S,1R,4R,5R)-4-[6-chloropurin-9-yl]-7,7-dimethyl-3,6,8-trioxabicyclo[-
3.3.0]octane-2-carbaldehyde (1 mmol) in tetrahydrofuran (20 ml),
and the mixture was stirred for 2 hours at -78.degree. C. The
reaction mixture was then allowed to warm to room temperature and
stirred for 24 hours, then quenched with aqueous ammonium chloride,
and partitioned between water and ethyl acetate. The organic layer
was separated, dried over magnesium sulfate, filtered, and the
solvent removed from the filtrate under reduced pressure, to yield
(1R,5R,6R,8R)-6-(6-chloropurin-9-yl)-8-ethynyl-3,3-dimethyl-2,4,7-trioxab-
icyclo[3.3.0]octane.
[0469] 2) To a solution of
(1R,5R,6R,8R)-6-(6-chloropurin-9-yl)-8-ethynyl-3,3-dimethyl-2,4,7-trioxab-
icyclo[3.3.0]octane (50 mg) and trans 2-aminocyclopentanol (0.04 g)
in ethanol was added triethylamine (0.12 ml), and the mixture was
stirred at 60.degree. C. for 32 hours. Solvent was then removed
under reduced pressure, and the residue dissolved in ethyl acetate,
washed with dilute nitric acid, followed by brine, dried over
sodium sulfate, filtered, and the solvent removed from the filtrate
under reduced pressure, to provide
2-[9-((1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]oct--
2-yl)purin-6-yl]cyclopentan-1-ol.
[0470] 3) In a sealed tube,
2-[9-((1R,2R,4R,5R)-4-ethynyl-7,7-dimethyl-3,6,8-trioxabicyclo[3.3.0]-oct-
-2-yl)purin-6-yl]cyclopentan-1-ol was stirred in 10 ml of 80%
acetic acid/water overnight. The solvent was then removed under
reduced pressure, and the residue purified by preparative thin
layer chromatography, eluting with 10% methanol/methylene chloride,
to provide
(4S,2R,3R,5R)-5-ethynyl-2-{6-[(2-hydroxycyclopentyl)amino]purin-9-yl}oxol-
ane-2,3-diol.
[0471] All compounds of Formula I were characterized by nmr spectra
and mass spectra. For example:
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-ethynyloxolane-3,4-diol
[0472] .sup.1H-NMR (CDCl.sub.3) 1.54-1.79 (m, 6H), 2.06-2.13 (m,
2H), 3.47 (s, 1H), 4.45-4.58 (m, 2H), 4.71 (s, 1H), 4.94 (s, 1H),
6.01 (d, 1H, J =5.09 Hz), 6.38 (s, 1H, NH), 8.03 (s, 1H), 8.26 (s,
1H). MH.sup.+ 317.
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-[2-(2-fluorophenyl)ethy-
nyl]oxolane-3,4-diol
[0473] .sup.1H-NMR (CDCl.sub.3) 1.51-1.77 (m, 6H), 2.08-2.14 (m,
2H), 4.03 (s, 1H, OH), 4.56 (s, 1H, OH), 4.61 (d, 1H, J=4.30 Hz),
4.78-4.81 (m, 1H), 5.21 (s, 1H), 5.94 (d, 1H, J=6.65 Hz), 6.07 (d,
1H, J=5.48 Hz), 6.56 (s, 1H, NH), 7.01-7.07 (m, 2H), 7.26-7.35 (m,
2H), 8.11 (s, 1H), 8.28 (s, 1H) MH.sup.+ 412.
EXAMPLE 18
[0474] Hard gelatin capsules containing the following ingredients
are prepared: TABLE-US-00001 Quantity Ingredient (mg/capsule)
Active Ingredient 30.0 Starch 305.0 Magnesium stearate 5.0
[0475] The above ingredients are mixed and filled into hard gelatin
capsules.
EXAMPLE 19
[0476] A tablet formula is prepared using the ingredients below:
TABLE-US-00002 Quantity Ingredient (mg/tablet) Active Ingredient
25.0 Cellulose, microcrystalline 200.0 Colloidal silicon dioxide
10.0 Stearic acid 5.0
[0477] The components are blended and compressed to form
tablets.
EXAMPLE 20
[0478] A dry powder inhaler formulation is prepared containing the
following components: TABLE-US-00003 Ingredient Weight % Active
Ingredient 5 Lactose 95
[0479] The active ingredient is mixed with the lactose and the
mixture is added to a dry powder inhaling appliance.
EXAMPLE 21
[0480] Tablets, each containing 30 mg of active ingredient, are
prepared as follows: TABLE-US-00004 Quantity Ingredient (mg/tablet)
Active Ingredient 30.0 mg Starch 45.0 mg Microcrystalline cellulose
35.0 mg Polyvinylpyrrolidone 4.0 mg (as 10% solution in sterile
water) Sodium carboxymethyl starch 4.5 mg Magnesium stearate 0.5 mg
Talc 1.0 mg Total 120 mg
[0481] The active ingredient, starch and cellulose are passed
through a No. 20 mesh U.S. sieve and mixed thoroughly. The solution
of polyvinylpyrrolidone is mixed with the resultant powders, which
are then passed through a 16 mesh U.S. sieve. The granules so
produced are dried at 50.degree. C. to 60.degree. C. and passed
through a 16 mesh U.S. sieve. The sodium carboxymethyl starch,
magnesium stearate, and talc, previously passed through a No. 30
mesh U.S. sieve, are then added to the granules which, after
mixing, are compressed on a tablet machine to yield tablets each
weighing 120 mg.
EXAMPLE 22
[0482] Suppositories, each containing 25 mg of active ingredient
are made as follows: TABLE-US-00005 Ingredient Amount Active
Ingredient 25 mg Saturated fatty acid glycerides to 2,000 mg
[0483] The active ingredient is passed through a No. 60 mesh U.S.
sieve and suspended in the saturated fatty acid glycerides
previously melted using the minimum heat necessary. The mixture is
then poured into a suppository mold of nominal 2.0 g capacity and
allowed to cool.
EXAMPLE 23
[0484] Suspensions, each containing 50 mg of active ingredient per
5.0 mL dose are made as follows: TABLE-US-00006 Ingredient Amount
Active Ingredient 50.0 mg Xanthan gum 4.0 mg Sodium carboxymethyl
cellulose (11%) Microcrystalline cellulose (89%) 50.0 mg Sucrose
1.75 g Sodium benzoate 10.0 mg Flavor and Color q.v. Purified water
to 5.0 mL
[0485] The active ingredient, sucrose and xanthan gum are blended,
passed through a No. 10 mesh U.S. sieve, and then mixed with a
previously made solution of the microcrystalline cellulose and
sodium carboxymethyl cellulose in water. The sodium benzoate,
flavor, and color are diluted with some of the water and added with
stirring. Sufficient water is then added to produce the required
volume.
EXAMPLE 24
[0486] A subcutaneous formulation may be prepared as follows:
TABLE-US-00007 Ingredient Quantity Active Ingredient 5.0 mg Corn
Oil 1.0 mL
EXAMPLE 25
[0487] An injectable preparation is prepared having the following
composition: TABLE-US-00008 Ingredients Amount Active ingredient
2.0 mg/ml Mannitol, USP 50 mg/ml Gluconic acid, USP q.s. (pH 5-6)
water (distilled, sterile) q.s. to 1.0 ml Nitrogen Gas, NF q.s.
EXAMPLE 26
[0488] A topical preparation is prepared having the following
composition: TABLE-US-00009 Ingredients grams Active ingredient
0.2-10 Span 60 2.0 Tween 60 2.0 Mineral oil 5.0 Petrolatum 0.10
Methyl paraben 0.15 Propyl paraben 0.05 BHA (butylated hydroxy
anisole) 0.01 Water q.s. to 100
[0489] All of the above ingredients, except water, are combined and
heated to 60.degree. C. with stirring. A sufficient quantity of
water at 60.degree. C. is then added with vigorous stirring to
emulsify the ingredients, and water then added q.s. 100 g.
EXAMPLE 27
[0490] TABLE-US-00010 Sustained Release Composition Weight
Preferred Ingredient Range (%) Range (%) Most Preferred Active
ingredient 50-95 70-90 75 Microcrystalline cellulose (filler) 1-35
5-15 10.6 Methacrylic acid copolymer 1-35 5-12.5 10.0 Sodium
hydroxide 0.1-1.0 0.2-0.6 0.4 Hydroxypropyl methylcellulose 0.5-5.0
1-3 2.0 Magnesium stearate 0.5-5.0 1-3 2.0
[0491] The sustained release formulations of this invention are
prepared as follows: compound and pH-dependent binder and any
optional excipients are intimately mixed(dry-blended). The
dry-blended mixture is then granulated in the presence of an
aqueous solution of a strong base which is sprayed into the blended
powder. The granulate is dried, screened, mixed with optional
lubricants (such as talc or magnesium stearate), and compressed
into tablets. Preferred aqueous solutions of strong bases are
solutions of alkali metal hydroxides, such as sodium or potassium
hydroxide, for example sodium hydroxide, in water (optionally
containing up to 25% of water-miscible solvents such as lower
alcohols).
[0492] The resulting tablets may be coated with an optional
film-forming agent, for identification, taste-masking purposes and
to improve ease of swallowing. The film forming agent will
typically be present in an amount ranging from between 2% and 4% of
the tablet weight. Suitable film-forming agents are well known to
the art and include hydroxypropyl. methylcellulose, cationic
methacrylate copolymers (dimethylaminoethyl
methacrylate/methyl-butyl methacrylate copolymers--Eudragit.RTM.
E--Rohm. Pharma), and the like. These film-forming agents may
optionally contain colorants, plasticizers, and other supplemental
ingredients.
[0493] The compressed tablets have a hardness sufficient to
withstand 8 Kp compression. The tablet size will depend primarily
upon the amount of compound in the tablet. The tablets will include
from 300 to 1100 mg of compound free base. For example, the tablets
will include amounts of compound free base ranging from 400-600 mg,
650-850 mg, and 900-1100 mg.
[0494] In order to influence the dissolution rate, the time during
which the compound containing powder is wet mixed is controlled.
For example the total powder mix time, i.e. the time during which
the powder is exposed to sodium hydroxide solution, will range from
1 to 10 minutes and preferably from 2 to 5 minutes. Following
granulation, the particles are removed from the granulator and
placed in a fluid bed dryer for drying at about 60.degree. C.
EXAMPLE 28
Materials
[0495] The A.sub.1-adenosine antagonists
8-cyclopentyl-1,3-dipropylxanthine (CPX) and
8-cyclopentyl-1,3-dimethylxanthine (CPT), the A.sub.1-adenosine
agonists N.sub.6-cyclopentyladenosine (CPA),
2-chloro-N.sub.6-cyclopentyladenosine (CCPA),
and-N.sub.6-cyclohexyladenosine (CHA), the adenosine deaminase
inhibitor erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), the
adenosine kinase inhibitor iodotubercidin, and forskolin were
purchased from Research Biochemicals (Natick, Mass.).
{[(5-{6-[(3R)oxolan-3-yl]amino}purin-9-yl)(3S,2R,4R)-3,4-di-hydroxyoxolan-
-2-yI]methoxy}-N-methylcarboxamide, molecular weight 394.38, is a
derivative of the selective A.sub.1-adenosine receptor full agonist
CVT-510. Adenosine was purchased from Sigma Chemical (St. Louis,
Mo.). The radioligand
8-cyclopentyl-1,3-dipropyl-[2,3-.sup.3H(N)]xanthine ([.sup.3H]CPX)
was purchased from New England Nuclear (Boston, Mass.).
Concentrated stock solutions (10-100 mM) of CVT-2759, CPX, CPT,
CPA, CCPA, CHA, and forskolin were dissolved in dimethylsulfoxide,
stored as aliquots at -80.degree. C., and diluted in physiological
saline for use in experiments. The final content of
dimethylsulfoxide in saline during experiments was not more than
0.1%. Adenosine and EHNA were dissolved in saline immediately
before use.
Binding Assays--DDT.sub.1 Cells
Cell Culture
[0496] DDT cells (hamster vas deferens smooth muscle cell line)
were grown as monolayers in petri dishes using Dulbecco's Modified
Eagle's Medium (DMEM) containing 2.5 .mu.g ml.sup.-1 amphotericin
B, 100 U ml.sup.-1 penicillin G, 0.1 mg ml.sup.-1 streptomycin
sulfate and 5% fetal bovine serun in a humidified atmosphere of 95%
air and 5% CO.sub.2. Cells were subcultured twice weekly by
dispersion in Hank's Balanced Salt Solution (HBSS) without the
divalent cations and containing 1 mM EDTA. The cells were then
seeded in growth medium at a density of 1.2.times.10.sup.5 cells
per plate and experiments were performed 4 days later at
approximately one day preconfluence.
Membrane Preparations
[0497] Cell layers were washed twice with HBSS (2.times.10 ml),
scraped free of the plate with the aid of a rubber policeman in 5
ml of 50 mM Tris-HCl buffer pH 7.4 at 4.degree. C. and the
suspension homogenized for 10 s. The homogenate was centrifuged at
27,000.times.g for 10 min, resuspended in buffer, and centrifuged
again, as described above. The protein content was determined with
a Biorad Protein Assay Kit (Richmond, Calif.) using bovine serum
albumin as standard. This membrane suspension was stored
dimethylsulfoxide (DMSO) in He buffer (10 mM HEPES, 1 uM EDTA at pH
7.4) and stored in liquid nitrogen at -80.degree. C.
Competitive Binding Assays:
[0498] Compounds of Formula I were assayed to determine their
affinity for the A.sub.1 adenosine receptor sites on the membranes
of DDT cells. Briefly, 50-70 ug of membrane protein were incubated
in a mixture containing 2 U/ml adenosine deaminase, 10 mM
GTP-.gamma.S in 5 mM HE buffer containing 5mM MgCl.sub.2 in glass
tubes. Stock solutions of the compounds of the invention were
serially diluted (10-.sup.10M to 10.sup.-4M) in HE buffer or HE
buffer alone (control to determine non-specific binding) and added
to the incubation mixture. Finally, tritiated cyclopentyladenosine
(.sup.3H-CPA) was added to a final concentration of 1.5 nM. After
incubation at 23.degree. C. 90 minutes, the reaction was stopped by
filtration on a Brandel MR24 cell harvester and washing with
ice-cold Tris-EDTA buffer (three times, approximate volume 10
ml/wash) over Whatman GF/B filters (presoaked for 1 h in 0.3%
polyethylenimine to reduce non-specific binding). Filters were
transferred to scintillation vials and 5 ml of Scintisafe (VWR,
Brisbane, Calif.) was added. The amount of radioactivity retained
on the filters was determined by liquid scintillation spectrometry.
Protein determinations were by the method of Bradford (1976. Anal.
Biochem. 72:248) using bovine serum albumin as the standard.
[0499] The compounds of Formula I were shown to be
A.sub.1-adenosine receptor agonists in this assay.
EXAMPLE 29
[.sup.35S]GTP.gamma.S Binding Assays
[0500] The ability of agonists to activate G proteins was
determined by using radiolabeled GTP ([.sup.35S]GTP.gamma.S).
Briefly, membrane proteins (30-50 .mu.g/assay tube) were placed in
glass tubes containing 50 mM Tris-HCl buffer pH 7.4, 5 mM
MgCl.sub.2, 100 mM NaCl, 1 mM dithiothreitol, 0.2 units ml.sup.-1
adenosine deaminase, 0.5% BSA, 1 mM EDTA, 10 mM GDP, and 0.3 nM
[.sup.35S]GTP.gamma.S. Varying concentrations of the compounds of
the invention (putative A.sub.1 adenosine receptor agonists), a
known A.sub.1 adenosine receptor full agonist N
cyclopentlyladenosine (CPA or CCPA) or a control tube containing 10
uM GTP.gamma.S but no agonist (to determine nonspecific binding)
were added to separate tubes. The assay tubes were incubated for 90
minutes at 37.degree. C. Agonist stimulated binding was assessed by
determining the difference between total binding in the presence of
putative agonists and basal binding determined in the absence of
CPA. Results were expressed as the percentage stimulation of the
putative agonists relative to the full agonist CPA after
subtracting out non-specific binding.
[0501] The compounds of Formula I were shown to be
A.sub.1-adenosine receptor agonists in this assay.
EXAMPLE 30
Guinea Pig Isolated Perfused Hearts
[0502] Guinea pigs (Hartley) of either sex weighing 300-350 g are
anaesthetized with methoxyflurane and killed by decapitation. The
chest is cut open, and the heart quickly removed and rinsed in
ice-cold modified Krebs-Henseleit (K-H) solution. The contents of
the modified K-H solution are (in mM) 117.9 NaCl, 4.8 KCl, 2.5
CaCl.sub.2, 1.18 MgSO.sub.2, 1.2 KH.sub.2PO.sub.4, 0.5 Na.sub.2
EDTA, 0.14 ascorbic acid, 5.5 dextrose, 2.0 pyruvic acid (sodium
salt), and 25 NaHCO.sub.3. The K-H solution is continuously gassed
with 95% 0.sub.2-5% C0.sub.2, and the pH adjusted to a value of
7.4. To perfuse the heart by the Langendorff method, the transected
aorta is put onto a glass cannula and secured by a ligature.
Retrograde perfusion of the aorta is initiated immediately at a
constant flow of 10 ml/min with modified K-H solution warmed to
36.0.+-.0.5.degree. C. A side port in the cannula is used to
connect the perfusion line to a Gould pressure transducer for
measurement of coronary perfusion pressure. Coronary perfusion
pressure was continuously recorded on a strip chart (Gould RS3400,
Cleveland, Ohio) throughout each experiment. Coronary conductance
(in mlmin-.sup.1mmHg-1) is calculated as the ratio of coronary flow
(10 ml/min) to perfusion pressure (in mmHg). To facilitate the exit
of fluid from the left ventricle, the leaflets of the mitral valve
are trimmed with fine spring-handled scissors. When appropriate,
hearts are paced at a constant rate using external electrodes.
After completion of dissection and instrumentation, stimulus-to-His
bundle (S-H) interval and coronary perfusion pressure is monitored
continuously, each heart being allowed to equilibrate for 20-40 min
before the administration of drug. Experimental interventions are
always preceded and followed by control measurements. Criteria for
the exclusion of hearts from the study are 1) a coronary perfusion
pressure of <50 mmHg, 2) absence of a stable coronary perfusion
pressure during the equilibration period, and 3) inability to pace
a heart at a constant rate throughout an experiment.
[0503] For electrical pacing of hearts, a bipolar Teflon-coated
electrode is placed in the wall of the intra-atrial septum. Parts
of the left and right atrial tissues, including the region of the
sinoatrial node, are removed, both to decrease the spontaneous
heart rate and to expose the atrial septum for electrode placement.
Hearts are electrically paced at a fixed rate of 3.2 HZ. Stimuli
are provided by an interval generator (model 1830, WPI, Sarasota,
Fla.) and delivered through a stimulus isolation unit (model 1880,
WPI) as square wave pulses of 3 ms in duration and at least twice
the threshold intensity.
[0504] S-H interval. Prolongation of the S-H interval is used as a
measure of the negative dromotropic effect of A.sub.1-adenosine
agonists on AV nodal conduction. The His bundle electrogram is
recorded from a unipolar electrode placed in the right side of the
interatrial septum adjacent to the AV junction. The signal is
displayed continuously in real time on an oscilloscope screen at a
sweep rate of 10 ms/cm. The duration of time from the first pacing
artifact to the maximum upward deflection of the His bundle signal
is used as the S-H interval.
[0505] Hearts are equilibrated until the S-H interval and CPP
remains constant. The test compound is used to the perfused line in
a final concentration of 0.3, 3, 10 and in some hearts up to 30
.mu.M. If the second degree AV block happens at any concentration
before 30 .mu.M, the test compound is withdrawn to washout. After
washout of the first test compound, a second test compound could
not be used in the same heart unless the SH interval and CPP comes
back to the control or S-H interval is prolonged less than 2 ms
compared to the control. Up to three compounds can be used in the
same heart.
[0506] The compounds of Formula I demonstrate the ability to delay
AV nodal conduction in this assay.
EXAMPLE 31
Anti-Emesis Studies
[0507] For all the experiments adult male ferrets, body weight
range 1-1.5kg, are used. Emesis is induced by X-irradiation,
morphine and cisplatin.
X-Irradiation:
[0508] The ferrets are weighed on the day before the experiment. On
the day of the experiment each ferret receives 2 Gy (200 Rad) whole
body X-irradiation, administered over a 5 minute period. The
A.sub.1 receptor agonists or partial agonists are administered via
the subcutaneous route immediately after X-irradiation(i.e.
approximately 25 minutes before the onset of emesis). When
investigating the effects of the A1 receptor antagonist, DPCPX, on
the partial agonist either both compounds are administered
simultaneously immediately after X-irradiation, or the DPCPX was
given as a 15 minute pre-treatment. In all cases, the ferrets are
observed for 2 hours after X-irradiation, and the time and numbers
of retches and vomits are recorded.
Morphine:
[0509] The ferrets are weighed on the day before the experiment.
The adenosine A.sub.1 receptor agonist is administered
subcutaneously 15 minutes before the 0.5 mg kg-1 subcutaneous dose
of morphine. (Emesis normally starts 5 minutes after morphine
administration). The ferrets are observed for 2 hours after the
morphine dose and the time and numbers of retches and vomits are
recorded.
Cisplatin:
[0510] The ferrets are weighed and measured for the calculation of
body surface area on the day before the experiment. On the day of
the experiment each ferret receives an intraperitoneal dose of
cisplatin (200 mg m-2). The adenosine A.sub.1 receptor agonist is
administered subcutaneously immediately after the first emetic
episode (approximately 1.5 hours after cisplatin administration).
The ferrets are observed for 7 hours after the first emetic episode
and the time and numbers of retches and vomits are recorded.
[0511] The compounds of Formula I demonstrate the ability to
control emesis in this assay.
Determination of Antilipolyitic Properties
[0512] Animals. Male Sprague-Dawley rats (380-420 g) were purchased
from Simonsen Laboratories (Gilroy, Calif.). All animals received
humane care according to the guidelines set forth in The Principles
of Laboratory Animal Care formulated by the National Society for
Medical Research and the Guide for the Care and Use of Laboratory
Animals prepared by the Institute of Laboratory Animal Resources
and published by the National Institute of Health (NIH Publication
86-23, revised 1996).
[0513] Isolation of Rat Epididymal Adipocytes. Adipocytes were
isolated from the epididymal fat pads of rats as described
previously (Rodbell, 1964). Briefly, rats were anesthetized using
methoxyfluorane and killed by exsanguination. Epididymal fat tissue
was removed and placed into a modified Krebs (KRH) solution
containing NaCl (100 mM), KCl (4.7 mM), CaCl2 (2.5 mM), NaHCO3 (3.6
mM), MgSO4 (1.19 mM), KH2PO4 (1.18 mM), dextrose (5 mM), pyruvic
acid (5 mM), ascorbic acid (1 mM), and HEPES (5 mM), pH 7.4.
Visible blood vessels were dissected and excised, and the adipose
tissue was minced. Minced tissue was digested with 25 ml of fresh
KRH solution containing type I collagenase (1 mg/ml), fatty-acid
free BSA [1%(wt/vol)] and nicotinic acid (2_M, to inhibit
lipolysis) for 40 to 60 minutes at 37.degree. C. with continuous
gentle shaking. The cell suspension was filtered through a
nylon-mesh (210_m) to remove undigested tissue fragments. The cell
filtrate was washed three times using KRH solution containing 1%
fatty acid-free BSA at 37.degree. C. The final adipocyte suspension
was either diluted in fresh KRH solution with 1% fatty acid-free
BSA for use in cAMP experiments, or used to prepare membranes for
radioligand binding assays.
[0514] cAMP Assays in Isolated Rat Adipocytes. Aliquots (100 .mu.l,
45,000-90,000 cells) of the freshly prepared adipocyte cell
suspension were placed into wells of 24-well cell culture plates
containing 0.4 ml of KRH solution containing fatty acid-free BSA
(1%), ascorbic acid (1 mM), rolipram (10 .mu.M), cilostamide (1
.mu.M), adenosine deaminase (2 U/ml), and appropriate A.sub.1
adenosine receptor agonist(s). An aliquot of 0.5 ml of KRH solution
containing 60 nM isoproterenol was added to each well, and
incubations proceeded for 4 min in an orbital shaker bath
maintained at 37.degree. C. Assays were terminated by the addition
of 200 .mu.l of 300 mM HCl to each well to lyse the cells. The
concentration of cAMP in the cell lysate was determined using
colorimetric direct cAMP kits (Assay Designs, Inc., Ann Arbor,
Mich.).
Adipocyte Membrane Preparation and Competition Binding
[0515] Assays. Freshly isolated adipocytes were added to a chilled
solution containing sucrose (0.25 M), EDTA (1 mM), and Tris-HCl (10
mM, pH 7.4) and homogenized with 10 strokes using a motordriven
tissue grinder. The homogenate was cooled on ice and the fat layer
was discarded. The homogenate was then centrifuged at 500 g for 10
min at 4.degree. C. The supernatant under the fat layer was
removed, resuspended in fresh buffer, and homogenized a second time
with six strokes using the tissue grinder. Cell membranes were
collected by centrifugation of the homogenate at 15,000 g for 15
min. The final membrane pellet was resuspended in a solution
containing sucrose (0.25 M), phenylmethylsulfonyl fluoride (0.1
mM), leupeptin (5 .mu.g/ml), aprotinin (5 g/ml), adenosine
deaminase (2 U/ml), and Tris-HCl (10 mM) buffer, pH 7.4. The
membrane suspension was frozen and stored in liquid nitrogen. For
competition binding assays, membrane suspensions were thawed and
incubated for 2 hours at room temperature in Tris-HCl (50 mM)
buffer containing ADA (1 U/ml), guanosine 5-(imido)triphosphate
(100 M), and [3H]CPX (1-3nM) and progressively higher
concentrations of the competing agent. At the end of incubation,
free radioligand was separated from membrane-bound radioligand by
filtration through GF/C glass fiber filters (Whatman, Maidstone,
UK) using a tissue harvester (Brandel, Inc., Gaithersburg, Md.).
Radioactivity was quantified by liquid scintillation counting.
Nonspecific binding of [3H]CPX was defined as [3H]CPX bound in the
presence of 10 .mu.MN6-cyclopentyltheophylline. Triplicate
determinations were performed for each concentration of unlabeled
compounds.
Effects of Compounds of Formula I on Heart Rate and Serum NEFA
Concentration in Awake Rats.
[0516] Heart rate was measured from rats chronically instrumented
with telemetry transmitters. For transmitter implantation, a
midline laparotomy was performed on anesthetized rats and a
transmitter for ECG recording was sutured to the abdominal wall.
The two electrocardiographic leads were tunneled through the wall,
passed subcutaneously (one to the left shoulder, the other to the
right thigh), and secured in place with sutures. Heart rates of
awake rats were measured using a Dataquest ART Gold System (Data
Sciences International, St. Paul, Minn.). Cardiac electrical
activity was recorded for 10-s periods and used to calculate heart
rate in beats per minute. After recording of a baseline heart rate,
either vehicle (0.9% DMSO in saline, 0.5 ml) or a compound of
Formula I was injected into the intraperitoneal cavity of each rat,
and heart rate was monitored at intervals for an additional 3
hours.
[0517] The effects of a compound of Formula I on heart rate and
serum NEFA reduction concentration were determined in separate
groups of rats to avoid the effects of animal handling and blood
sampling on heart rate. Three days before an experiment, a catheter
(0.025-mm outer diameter) was implanted in the left common carotid
artery of each rat using aseptic conditions and sterile technique.
The catheter was tunneled subcutaneously to the dorsal surface.
After recovery from anesthesia, rats were placed in metabolic cages
to facilitate handling and blood sampling. Blood samples (0.2 ml)
were drawn before and at various time points after i.p. injection
of either a compound of Formula I or vehicle (DMSO in saline). A
0.4-ml volume of 1% sodium citrate in saline was administered after
withdrawal of each blood sample to replace blood volume and prevent
clotting in the carotid artery catheter. Serum was collected from
each sample after centrifugation of the clotted blood. Serum
samples were stored at .sub.--80.degree. C. until analysis. Serum
NEFA concentration was determined using an enzymatic colorimetric
assay kit (Wako Chemicals, Richmond, Va.).
[0518] The antilipolytic properties of compounds of Formula I in
rats with catecholamines-stimulated NEFA levels were studied in
rats with indwelling catheters (described above). Norepinephrine
was delivered either by i.v. infusion into the jugular vein at 3
g/kg/min for 30 minutes or by i.p. injection (60.mu.g/kg). A
compound of Formula I was delivered by i.p. injection either before
or after norepinephrine to determine whether the increase in NEFA
concentrations caused by norepinephrine could be prevented or
reversed by the compound of Formula I.
[0519] The compounds of Formula I demonstrated antilipolytic
properties in this assay. For example, oral administration of
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{2-[2-fluorophenyl]-et-
hynyl}oxolane-3,4-diol at a dose level of 1 mg/Kg provided an
initial 40% reduction of non-esterified free fatty acid (NEFA) that
was maintained for 1 hour, after which time the plasma levels of
NEFA returned to normal in 2 hours. Oral administration of
(4S,2R,3R,5R)-2-[6-(cyclopentylamino)purin-9-yl]-5-{2-[2-fluorophenyl]-et-
hynyl}oxolane-3,4-diol at a dose level of 2.5 mg/Kg provided an
initial 60% reduction of non-esterified free fatty acid (NEFA) that
was maintained for 90 minutes, after which time the plasma levels
of NEFA returned to normal in 4 hours.
[0520] At dose levels of 1 mg/Kg, 2.5 mg/Kg, and 5 mg/Kg, no effect
on heart rate was observed.
* * * * *