U.S. patent application number 11/593381 was filed with the patent office on 2007-08-02 for novel 2'-c methyl nucleoside derivatives.
This patent application is currently assigned to Metabasis Therapeutics, Inc.. Invention is credited to Mark D. Erion, Malcolm MacCoss, David B. Olsen, K. Raja Reddy.
Application Number | 20070179114 11/593381 |
Document ID | / |
Family ID | 34841195 |
Filed Date | 2007-08-02 |
United States Patent
Application |
20070179114 |
Kind Code |
A1 |
Erion; Mark D. ; et
al. |
August 2, 2007 |
Novel 2'-C methyl nucleoside derivatives
Abstract
Compounds of Formula I, stereoisomers, and pharmaceutically
acceptable salts or prodrugs thereof, their preparation, and their
uses for the treatment of hepatitis C viral infection are
described: ##STR1##
Inventors: |
Erion; Mark D.; (Del Mar,
CA) ; Reddy; K. Raja; (San Diego, CA) ;
MacCoss; Malcolm; (Freehold, NJ) ; Olsen; David
B.; (Lansdale, PA) |
Correspondence
Address: |
WOLF GREENFIELD & SACKS, P.C.
600 ATLANTIC AVENUE
BOSTON
MA
02210-2206
US
|
Assignee: |
Metabasis Therapeutics,
Inc.
La Jolla
CA
Merck & Co., Inc.
Rahway
CA
|
Family ID: |
34841195 |
Appl. No.: |
11/593381 |
Filed: |
November 6, 2006 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
10903215 |
Jul 29, 2004 |
|
|
|
11593381 |
Nov 6, 2006 |
|
|
|
60544743 |
Feb 13, 2004 |
|
|
|
Current U.S.
Class: |
514/45 ; 514/47;
536/26.11; 536/26.12; 536/26.13 |
Current CPC
Class: |
A61P 31/18 20180101;
A61P 31/14 20180101; C07H 19/20 20130101; C07H 19/04 20130101; A61P
31/12 20180101 |
Class at
Publication: |
514/045 ;
536/026.11; 536/026.12; 536/026.13; 514/047 |
International
Class: |
A61K 31/7076 20060101
A61K031/7076; C07H 19/20 20060101 C07H019/20 |
Claims
1. A compound of Formula I: ##STR161## wherein: B is selected from
the group consisting of ##STR162## V is selected from the group
consisting of optionally substituted monocyclic aryl and optionally
substituted monocyclic heteroaryl; W and W' are independently
selected from the group consisting of --R.sup.2, optionally
substituted monocyclic aryl, and optionally substituted monocyclic
heteroaryl; Z is selected from the group consisting of halogen,
--CN, --COR.sup.5, --CONR.sup.4.sub.2, --CO.sub.2R.sup.5,
--SO.sub.2R.sup.5, --SO.sub.2NR.sup.4.sub.2, --OR.sup.4,
--SR.sup.4, --R.sup.4, --NR.sup.4.sub.2, --OCOR.sup.5,
--OCO.sub.2R.sup.5, --SCOR.sup.5, --SCO.sub.2R.sup.5,
--NHCOR.sup.4, --NHCO.sub.2R.sup.5, --(CH.sub.2).sub.p--OR.sup.6,
and --(CH.sub.2).sub.p--SR.sup.6; or together V and Z are connected
via an additional 3-5 atoms to form a cyclic group, optionally
containing 1 heteroatom, that is fused to an aryl group at the beta
and gamma position to the O attached to the phosphorus; or together
Z and W are connected via an additional 3-5 atoms to form a cyclic
group, optionally containing one heteroatom; or together W and W'
are connected via an additional 2-5 atoms to form a cyclic group,
optionally containing 0-2 heteroatoms; R.sup.2 is selected from the
group consisting of R.sup.3 and hydrogen; R.sup.3 is selected from
the group consisting of alkyl, aryl, heterocycloalkyl, and aralkyl;
R.sup.4 is selected from the group consisting of R.sup.3 and
hydrogen; R.sup.5 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl; R.sup.6 is selected from the
group consisting of hydrogen, and lower acyl; R.sup.12 is selected
from the group consisting of hydrogen, and lower acyl; and p is an
integer 2 or 3; or a pharmaceutically acceptable salt thereof.
2. The compound of claim 1 wherein: B is ##STR163##
3. The compound of claim 1 wherein: B is ##STR164##
4. (canceled)
5. A compound of Formula II: ##STR165## wherein: B is selected from
the group consisting of: ##STR166## V is selected from the group
consisting of optionally substituted monocyclic aryl and optionally
substituted monocyclic heteroaryl; W and W' are independently
selected from the group consisting of --H, methyl, and V, or W and
W' are each methyl, with the proviso that when W is V, then W' is
H; Z is selected from the group consisting of --H, --OMe, --OEt,
phenyl, C.sub.1-C.sub.3 alkyl, --NR.sup.4.sub.2, --SR.sup.4,
--(CH.sub.2).sub.p--OR.sup.6, --(CH.sub.2).sub.p--SR.sup.6 and
--OCOR.sup.5; or together V and Z are connected via an additional
3-5 atoms to form a cyclic group, optionally containing 1
heteroatom, that is fused to an aryl group at the beta and gamma
position to the O attached to the phosphorus; or together Z and W
are connected via an additional 3-5 atoms to form a cyclic group,
optionally containing one heteroatom; or together W and W' are
connected via an additional 2-5 atoms to form a cyclic group;
R.sup.4 is C.sub.1-C.sub.4 alkyl; R.sup.5 is selected from the
group consisting of C.sub.1-C.sub.4 alkyl, monocyclic aryl, and
monocyclic aralkyl; and R.sup.6 is C.sub.1-C.sub.4 acyl; R.sup.7
and R.sup.8 are independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 acyl, C.sub.1-C.sub.4 alkoxycarbonyl, and
a naturally-occurring L-amino acid connected via its carbonyl group
to form an ester; or together R.sup.7 and R.sup.8 form a cyclic
carbonate; R.sup.9 is selected from the group consisting of amino,
azido, --N.dbd.CHN(R.sup.4).sub.2, --NHC(O)R.sup.4, and
--NHC(O)OR.sup.4; and R.sup.10 is selected from the group
consisting of OR.sup.6, halogen, and H; or a pharmaceutically
acceptable salt thereof.
6. The compound of claim 1, wherein: V is selected from the group
consisting of phenyl, substituted phenyl with 1-3 substituents
independently selected from the group consisting of halogen,
C.sub.1-C.sub.6 alkyl, --CF.sub.3, --OR.sup.3, --OR.sup.12,
--COR.sup.3, --CO.sub.2R.sup.3, --NR.sup.3.sub.2,
--NR.sup.12.sub.2, --CO.sub.2NR.sub.2.sup.2, --SR.sup.3,
--SO.sub.2R.sup.3, --SO.sub.2NR.sub.2.sup.2 and --CN, monocyclic
heteroaryl, and substituted monocyclic heteroaryl with 1-2
substituents independently selected from the group consisting of
halogen, C.sub.1-C.sub.6 alkyl, --CF.sub.3, --OR.sup.3,
--OR.sup.12, --COR.sup.3, --CO.sub.2R.sup.3, --NR.sup.3.sub.2,
--NR.sup.12.sub.2, --CO.sub.2NR.sub.2.sup.2, --SR.sup.3,
--SO.sub.2R.sup.3, --SO.sub.2NR.sub.2.sup.2 and --CN, and wherein
said monocyclic heteroaryl and substituted monocyclic heteroaryl
has 1-2 heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that a) when there are
two heteroatoms and one is O, then the other can not be O or S, and
b) when there are two heteroatoms and one is S, then the other can
not be O or S; or together V and Z are connected via an additional
3-5 atoms to form a cyclic group, optionally containing 1
heteroatom, that is fused to an aryl group at the beta and gamma
position to the O attached to the phosphorus; and R.sup.3 is
C.sub.1-C.sub.6 alkyl.
7. The compound of claim 6 wherein: V is selected from the group
consisting of phenyl, substituted phenyl with 1-3 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN, monocyclic heteroaryl,
and substituted monocyclic heteroaryl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN and wherein said
monocyclic heteroaryl and substituted monocyclic heteroaryl has 1-2
heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that a) when there are
two heteroatoms and one is O, then the other can not be O or S, and
b) when there are two heteroatoms and one is S, then the other can
not be O or S; or together V and Z are connected via an additional
4 atoms to form a 6-membered ring that is fused to a phenyl or
substituted phenyl at the beta and gamma position to the O attached
to the phosphorus.
8. The compound of claim 7 wherein V is selected from the group
consisting of phenyl; substituted phenyl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; pyridyl; substituted
pyridyl with 1 substituent independently selected from the group
consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3; furanyl; substituted furanyl with 1 substituent
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; thienyl; and
substituted thienyl with 1 substituent independently selected from
the group consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3.
9. The compound of claim 8 wherein V is selected from the group
consisting of phenyl, 3-chlorophenyl, 3-bromophenyl, 2-bromophenyl,
3,5-dichlorophenyl, 3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl,
and 4-pyridyl.
10. The compound of claim 9 wherein V is selected from the group
consisting of 3-chlorophenyl, 3-bromophenyl, 2-bromophenyl,
3,5-dichlorophenyl, 3-pyridyl, and 4-pyridyl.
11. The compound of claim 1 wherein Z is selected from the group
consisting of --H, --OMe, --OEt, phenyl, C.sub.1-C.sub.3 alkyl,
--NR.sup.4.sub.2, --SR.sup.4, --(CH.sub.2).sub.p--OR.sup.6,
--(CH.sub.2).sub.p--SR.sup.6 and --OCOR.sup.5; R.sup.4 is
C.sub.1-C.sub.4 alkyl; R.sup.5 is selected from the group
consisting of C.sub.1-C.sub.4 alkyl, monocyclic aryl, and
monocyclic aralkyl; and R.sup.6 is C.sub.1-C.sub.4 acyl.
12. The compound of claim 11 wherein Z is selected from the group
consisting of --H, --OMe, --OEt, and phenyl.
13. The compound of claim 1 wherein: W and W' are independently
selected from the group consisting of --H, C.sub.1-C.sub.6 alkyl,
and phenyl; or together W and W' are connected via an additional
2-5 atoms to form a cyclic group.
14. The compound of claim 1 wherein W and W' are independently
selected from the group consisting of --H, methyl, and V, or W and
W' are each methyl, with the proviso that when W is V, then W' is
H.
15. The compound of claim 1 wherein: V is selected from the group
consisting of optionally substituted monocyclic aryl and optionally
substituted monocyclic heteroaryl; W and W' are independently
selected from the group consisting of --H, methyl, and V, or W and
W' are each methyl, with the proviso that when W is V, then W' is
H; Z is selected from the group consisting of --H, --OMe, --OEt,
phenyl, C.sub.1-C.sub.3 alkyl, --NR.sup.4.sub.2, --SR.sup.4,
--(CH.sub.2).sub.p--OR.sup.6, --(CH.sub.2).sub.p--SR.sup.6 and
--OCOR.sup.5; or together V and Z are connected via an additional
3-5 atoms to form a cyclic group, optionally containing 1
heteroatom, that is fused to an aryl group at the beta and gamma
position to the O attached to the phosphorus; or together Z and W
are connected via an additional 3-5 atoms to form a cyclic group,
optionally containing one heteroatom; or together W and W' are
connected via an additional 2-5 atoms to form a cyclic group; and
R.sup.4 is C.sub.1-C.sub.4 alkyl; R.sup.5 is selected from the
group consisting of C.sub.1-C.sub.4 alkyl, monocyclic aryl, and
monocyclic aralkyl; and R.sup.6 is C.sub.1-C.sub.4 acyl.
16. The compound of claim 15 wherein: V is selected from the group
consisting of phenyl, substituted phenyl with 1-3 substituents
independently selected from the group consisting of halogen,
C.sub.1-C.sub.6 alkyl, --CF.sub.3, --OR.sup.3, --OR.sup.12,
--COR.sup.3, --CO.sub.2R.sup.3, --NR.sup.3.sub.2,
--NR.sup.12.sub.2, --CO.sub.2NR.sub.2.sup.2, --SR.sup.3,
--SO.sub.2R.sup.3, --SO.sub.2NR.sub.2.sup.2 and --CN, monocyclic
heteroaryl, and substituted monocyclic heteroaryl with 1-2
substituents independently selected from the group consisting of
halogen, C.sub.1-C.sub.6 alkyl, --CF.sub.3, --OR.sup.3,
--OR.sup.12, --COR.sup.3, --CO.sub.2R.sup.3, --NR.sup.3.sub.2,
--NR.sup.12.sub.2, --CO.sub.2NR.sub.2.sup.2, --SR.sup.3,
--SO.sub.2R.sup.3, --SO.sub.2NR.sub.2.sup.2 and --CN, and wherein
said monocyclic heteroaryl and substituted monocyclic heteroaryl
has 1-2 heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that a) when there are
two heteroatoms and one is O, then the other can not be O or S, and
b) when there are two heteroatoms and one is S, then the other can
not be O or S; or together V and Z are connected via an additional
3-5 atoms to form a cyclic group, optionally containing 1
heteroatom, that is fused to an aryl group at the beta and gamma
position to the O attached to the phosphorus; and R.sup.3 is
C.sub.1-C.sub.6 alkyl.
17. The compound of claim 16 wherein: V is selected from the group
consisting of phenyl, substituted phenyl with 1-3 substituents
independently selected from the group consisting of Cl, --Br, --F,
C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN, monocyclic heteroaryl,
and substituted monocyclic heteroaryl with 1-2 substituents
independently selected from the group consisting of Cl, --Br, --F,
C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN, and wherein said
monocyclic heteroaryl and substituted monocyclic heteroaryl has 1-2
heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that a) when there are
two heteroatoms and one is O, then the other can not be O or S; and
b) when there are two heteroatoms and one is S, then the other can
not be O or S; or together V and Z are connected via an additional
4 atoms to form a 6-membered ring that is fused to a phenyl or
substituted phenyl at the beta and gamma position to the O attached
to the phosphorus.
18. The compound of claim 17 wherein V is selected from the group
consisting of phenyl; substituted phenyl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; pyridyl; substituted
pyridyl with 1 substituent independently selected from the group
consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3; furanyl; substituted furanyl with 1 substituent
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; thienyl; and
substituted thienyl with 1 substituent independently selected from
the group consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3.
19. The compound of claim 1 wherein: V is selected from the group
consisting of phenyl, 3-chlorophenyl, 3-bromophenyl, 2-bromophenyl,
3,5-dichlorophenyl, 3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl,
and 4-pyridyl; and Z is selected from the group consisting of --H,
OMe, OEt, and phenyl; and W and W' are independently selected from
the group consisting of --H and phenyl, or W and W' are each
methyl.
20. The compound of claim 1 wherein Z, W, and W' are each --H.
21. The compound of claim 1 wherein V and W are the same and each
is selected from the group consisting of optionally substituted
monocyclic aryl and optionally substituted monocyclic
heteroaryl.
22. The compound of claim 1 wherein: B is ##STR167## V is selected
from the group consisting of 3-chlorophenyl, 3-bromophenyl,
2-bromophenyl, 3,5-dichlorophenyl, 3-pyridyl, and 4-pyridyl; and Z,
W, and W' are each --H.
23. The compound of claim 22 wherein said compound is:
##STR168##
24. The compound of claim 1 wherein: B is ##STR169## V is selected
from the group consisting of 3-chlorophenyl, 3-bromophenyl,
2-bromophenyl, 3,5-dichlorophenyl, 3-pyridyl, and 4-pyridyl; and Z,
W, and W' are each --H.
25. The compound of claim 24 wherein said compound is:
##STR170##
26-28. (canceled)
29. The compound of claim 1 wherein said compound is a compound of
Formula V: ##STR171## wherein: V and the 5'oxymethylene group of
the ribose sugar moiety are cis to one another.
30. The compound of claim 5 wherein said compound is a compound of
Formula III: ##STR172## wherein: V and the 5'oxymethylene group of
the ribose sugar moiety are cis to one another.
31. The compound of claim 30 wherein V is selected from the group
consisting of phenyl, substituted phenyl with 1-3 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN, monocyclic heteroaryl,
and substituted monocyclic heteroaryl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN.
32. The compound of claim 31 wherein V is selected from the group
consisting of phenyl, 3-chlorophenyl, 3-bromophenyl, 2-bromophenyl,
3,5-dichlorophenyl, 3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl,
and 4-pyridyl.
33. The compound of claim 32 wherein said compound has
R-stereochemistry at the V-attached carbon and has
S-stereochemistry at the phosphorus center.
34. The compound of claim 32 wherein said compound has
S-stereochemistry at the V-attached carbon and has
R-stereochemistry at the phosphorus center.
35. The compound of claim 29 wherein V is selected from the group
consisting of phenyl, substituted phenyl with 1-3 substituents
independently selected from the group consisting of halogen,
C.sub.1-C.sub.6 alkyl, --CF.sub.3, --OR.sup.3, --OR.sup.12,
--COR.sup.3, --CO.sub.2R.sup.3, --NR.sup.3.sub.2,
--NR.sup.12.sub.2, --CO.sub.2NR.sub.2.sup.2, --SR.sup.3,
--SO.sub.2R.sup.3, --SO.sub.2NR.sub.2.sup.2 and --CN, monocyclic
heteroaryl, and substituted monocyclic heteroaryl with 1-2
substituents independently selected from the group consisting of
halogen, C.sub.1-C.sub.6 alkyl, --CF.sub.3, --OR.sup.3,
--OR.sup.12, --COR.sup.3, CO.sub.2R.sup.3, --NR.sup.3.sub.2,
--NR.sup.12.sub.2, --CO.sub.2NR.sub.2.sup.2, --SR.sup.3,
--SO.sub.2R.sup.3, --SO.sub.2NR.sub.2.sup.2 and --CN, and wherein
said monocyclic heteroaryl and substituted monocyclic heteroaryl
has 1-2 heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that a) when there are
two heteroatoms and one is O, then the other can not be O or S, and
b) when there are two heteroatoms and one is S, then the other can
not be O or S; and R.sup.3 is C.sub.1-C.sub.6 alkyl.
36. The compound of claim 35 wherein V is selected from the group
consisting of phenyl, substituted phenyl with 1-3 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN, monocyclic heteroaryl,
and substituted monocyclic heteroaryl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN and wherein said
monocyclic heteroaryl and substituted monocyclic heteroaryl has 1-2
heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that a) when there are
two heteroatoms and one is O, then the other can not be O or S, and
b) when there are two heteroatoms and one is S, then the other can
not be O or S; or together V and Z are connected via an additional
4 atoms to form a 6-membered ring that is fused to a phenyl or
substituted phenyl at the beta and gamma position to the O attached
to the phosphorus.
37. The compound of claim 36 wherein V is selected from the group
consisting of phenyl; substituted phenyl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; pyridyl; substituted
pyridyl with 1 substituent independently selected from the group
consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3; furanyl; substituted furanyl with 1 substituent
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; thienyl; and
substituted thienyl with 1 substituent independently selected from
the group consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3.
38. The compound of claim 37 wherein V is selected from the group
consisting of phenyl, 3-chlorophenyl, 3-bromophenyl, 2-bromophenyl,
3,5-dichlorophenyl, 3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl,
and 4-pyridyl.
39. The compound of claim 38 wherein V is selected from the group
consisting of 3-chlorophenyl, 3-bromophenyl, 2-bromophenyl,
3,5-dichlorophenyl, 3-pyridyl, and 4-pyridyl.
40. The compound of claim 29 wherein said compound has
R-stereochemistry at the V-attached carbon and has
S-stereochemistry at the phosphorus center.
41. The compound of claim 29 wherein said compound has
S-stereochemistry at the V-attached carbon and has
R-stereochemistry at the phosphorus center.
42. A pharmaceutical composition comprising a pharmaceutically
effective amount of a compound of claim 1 and a pharmaceutically
acceptable carrier.
43. A pharmaceutical composition comprising a pharmaceutically
effective amount of a compound of claim 5 and a pharmaceutically
acceptable carrier.
44-101. (canceled)
102. A compound of Formula VII: ##STR173## wherein B is selected
from the group consisting of: ##STR174## X is selected from the
group consisting of NH.sub.2, NHCH.sub.3, N(CH.sub.3).sub.2,
OCH.sub.3, SCH.sub.3, OH, and SH; Y and Y' are independently O or
NH; R.sup.14 is independently selected from the group consisting of
H and NH.sub.2; the heterocyclic base B may be further substituted
at any position on the heterocyclic base B with a substituent of a
molecular weight of less than 150 and selected from the group
consisting of halogen, alkyl, alkenyl, alkynyl, aryl, alkaryl,
cycloalkyl, acyl, and alkoxy, and wherein said substituents may be
coupled to the 6-position of the heterocyclic base via a carbon,
sulfur, oxygen, or selenium; V, W, and W' are independently
hydrogen, alkyl, alkenyl, alkynyl, aryl, alkaryl, each of which is
optionally substituted; and Z is hydrogen, CHWOH, CHWOCOW', SW, or
CH.sub.2aryl.
103-104. (canceled)
105. The compound of claim 102 wherein B is: ##STR175##
106. The compound of claim 105 wherein X is NH.sub.2.
Description
RELATED APPLICATIONS
[0001] This application is a divisional of U.S. application Ser.
No. 10/903,215 filed Jul. 29, 2004, which claims benefit of U.S.
Provisional Application No. 60/544,743 filed Feb. 13, 2004 and
which is incorporated by reference herein in its entirely,
including figures.
FIELD OF THE INVENTION
[0002] The present invention is directed towards novel 2'-C-methyl
nucleoside 5'-monophosphate derivatives, their preparation and
their uses. More specifically, the novel compounds are useful to
treat hepatitis C viral infections.
BACKGROUND
[0003] The following description of the background of the invention
is provided to aid in understanding the invention, but is not
admitted to be, or to describe, prior art to the invention. All
publications are incorporated by reference in their entirety.
[0004] Hepatitis C is a viral disease that causes inflammation of
the liver that may lead to cirrhosis, primary liver cancer and
other long-term complications. Nucleosides are a well-recognized
class of compounds shown to be effective against a variety of viral
infections, including hepatitis B, HIV, and herpes. A few
nucleosides are reported to inhibit hepatitis C (HCV) virus
replication, including ribavirin, which currently is marketed as a
drug combination with various interferons, and nucleosides
containing a 2'-C-methyl ribose sugar.
[0005] Nucleosides are generally effective as antiviral agents
following conversion of the nucleoside to the corresponding
nucleoside 5'-triphosphate (NTP). Conversion occurs inside cells
through the action of various intracellular kinases. The first
step, i.e. conversion of the nucleoside to the 5'-monophosphate
(NMP) is generally the slow step and involves a nucleoside kinase,
which is encoded by either the virus or host. Conversion of the NMP
to the NTP is generally catalyzed by host nucleotide kinases. The
NTP interferes with viral replication through inhibition of viral
polymerases and/or via incorporation into a growing strand of DNA
or RNA followed by chain termination.
[0006] Use of nucleosides to treat viral liver infections is often
complicated by one of two problems. In some cases, the desired
nucleoside is a good kinase substrate and accordingly produces NTP
in the liver as well as other cells and tissues throughout the
body. Since NTP production is often associated with toxicity,
efficacy can be limited by extrahepatic toxicities. In other cases,
the desired nucleoside is a poor kinase substrate so is not
efficiently converted into the NMP and ultimately into the NTP.
[0007] For instance, U.S. Pat. No. 6,312,662 discloses the use of
certain phosphate prodrugs for the liver-specific delivery of
various drugs including nucleosides for the treatment of patients
with liver diseases such as hepatitis C, hepatitis B and
hepatocellular carcinoma.
SUMMARY OF THE INVENTION
[0008] The present invention is directed towards novel 2'-C-methyl
nucleoside 5'-monophosphate derivatives, their preparation and
their uses for the treatment of hepatitis C viral infections.
[0009] In one aspect, the present invention relates to compounds of
Formula I, and pharmaceutically acceptable salts and prodrugs
thereof. ##STR2##
[0010] wherein:
[0011] B is selected from the group consisting of ##STR3##
[0012] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl;
[0013] W and W' are independently selected from the group
consisting of --R.sup.2, optionally substituted monocyclic aryl,
and optionally substituted monocyclic heteroaryl;
[0014] Z is selected from the group consisting of halogen, --CN,
--COR.sup.5, --CONR.sup.4.sub.2, --CO.sub.2R.sup.5,
--SO.sub.2R.sup.5, --SO.sub.2NR.sup.4.sub.2, --OR.sup.4,
--SR.sup.4, --R.sup.4, --NR.sup.4.sub.2, --OCOR.sup.5,
--OCO.sub.2R.sup.5, --SCOR.sup.5, --SCO.sub.2R.sup.5,
--NHCOR.sup.4, --NHCO.sub.2R.sup.5, --(CH.sub.2).sub.p--OR.sup.6,
and --(CH.sub.2).sub.p--SR.sup.6; or
[0015] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; or
[0016] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0017] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group, optionally containing 0-2 heteroatoms;
[0018] R.sup.2 is selected from the group consisting of R.sup.3 and
hydrogen;
[0019] R.sup.3 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0020] R.sup.4 is selected from the group consisting of R.sup.3 and
hydrogen;
[0021] R.sup.5 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0022] R.sup.6 is selected from the group consisting of hydrogen,
and lower acyl;
[0023] R.sup.12 is selected from the group consisting of hydrogen,
and lower acyl; and
[0024] p is an integer 2 or 3.
[0025] In another aspect, the invention relates to compounds of
Formula I, and pharmaceutically acceptable salts and prodrugs
thereof: ##STR4##
[0026] wherein:
[0027] B is ##STR5##
[0028] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl;
[0029] W and W' are independently selected from the group
consisting of --R.sup.2, optionally substituted monocyclic aryl,
and optionally substituted monocyclic heteroaryl;
[0030] Z is selected from the group consisting of halogen, --CN,
--COR.sup.5, --CONR.sup.4.sub.2, --CO.sub.2R.sup.5,
--SO.sub.2R.sup.5, --SO.sub.2NR.sup.4.sub.2, --OR.sup.4,
--SR.sup.4, --R.sup.4, --NR.sup.4.sub.2, --OCOR.sup.5,
--OCO.sub.2R.sup.5, --SCOR.sup.5, --SCO.sub.2R.sup.5,
--NHCOR.sup.4, --NHCO.sub.2R.sup.5, --(CH.sub.2).sub.p--OR.sup.6,
and --(CH.sub.2).sub.p--SR.sup.6; or
[0031] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; or
[0032] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0033] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group, optionally containing 0-2 heteroatoms;
[0034] R.sup.2 is selected from the group consisting of R.sup.3 and
hydrogen;
[0035] R.sup.3 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0036] R.sup.4 is selected from the group consisting of R.sup.5 and
hydrogen;
[0037] R.sup.5 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0038] R.sup.6 is selected from the group consisting of hydrogen,
and lower acyl;
[0039] R.sup.12 is selected from the group consisting of hydrogen,
and lower acyl; and
[0040] p is an integer 2 or 3.
[0041] Some of the compounds of Formula I have asymmetric centers
where the stereochemistry is unspecified, and the diastereomeric
mixtures of these compounds are included, as well as the individual
stereoisomers when referring to a compound of Formula I
generally.
[0042] Some of the compounds described herein may exist as
tautomers such as keto-enol tautomers and imine-enamine tautomers.
The individual tautomers as well as mixtures thereof are
encompassed with compounds of Formula I. An example of keto-enol
tautomers which are intended to be encompassed within the compounds
of the present invention is illustrated below: ##STR6##
[0043] An example of imine-enamine tautomers which are intended to
be encompassed within the compounds of the present invention is
illustrated below: ##STR7##
[0044] Also provided are pharmaceutical compositions comprising
compounds of Formula I, pharmaceutically acceptable salts or
prodrugs thereof; in association with pharmaceutically acceptable
excipients or carriers.
[0045] Also provided are methods for inhibiting viral replication
comprising the step of administering to a patient a therapeutically
effective amount of a compound of Formula I, pharmaceutically
acceptable salts or prodrugs thereof.
[0046] Also provided are methods for inhibiting RNA-dependent RNA
viral replication comprising the step of administering to a patient
a therapeutically effective amount of a compound of Formula I, or
pharmaceutically acceptable salts or prodrugs thereof.
[0047] Also provided are methods for inhibiting HCV replication
comprising the step of administering to a patient a therapeutically
effective amount of a compound of Formula I, pharmaceutically
acceptable salts or prodrugs thereof.
[0048] Also provided are methods for treating viral infections
comprising the step of administering to a patient a therapeutically
effective amount of a compound of Formula I, or pharmaceutically
acceptable salts or prodrugs thereof.
[0049] Also provided are methods for treating viral infections of
the liver comprising the step of administering to a patient a
therapeutically effective amount of a compound of Formula I,
or pharmaceutically acceptable salts or prodrugs thereof.
[0050] Also provided are methods for treating RNA-dependent RNA
viral infection comprising the step of administering to a patient a
therapeutically effective amount of a compound of Formula I, a
pharmaceutically acceptable salts or prodrugs thereof.
[0051] Also provided are methods for treating HCV infection
comprising the step of administering to a patient a therapeutically
effective amount of a compound of Formula I, pharmaceutically
acceptable salts or prodrugs thereof.
[0052] Also provided are methods for preparing compounds of Formula
I, stereoisomers, and pharmaceutically acceptable salts or prodrugs
thereof.
DEFINITIONS
[0053] In accordance with the present invention and as used herein,
the following terms are defined with the following meanings, unless
explicitly stated otherwise.
[0054] The term "alkyl" refers to saturated aliphatic groups
including straight-chain, branched chain and cyclic groups, up to
and including 10 carbon atoms. Suitable alkyl groups include
methyl, ethyl, n-propyl, isopropyl, and cyclopropyl. The alkyl may
be optionally substituted with 1-3 substituents.
[0055] The term "aryl" refers to aromatic groups which have 5-14
ring atoms and at least one ring having a conjugated pi electron
system and includes carbocyclic aryl, heterocyclic aryl and biaryl
groups, all of which may be optionally substituted. The aryl group
may be optionally substituted with 1-6 substituents.
[0056] Carbocyclic aryl groups are groups which have 6-14 ring
atoms wherein the ring atoms on the aromatic ring are carbon atoms.
Carbocyclic aryl groups include monocyclic carbocyclic aryl groups
and polycyclic or fused compounds such as optionally substituted
naphthyl groups.
[0057] Heterocyclic aryl or heteroaryl groups are groups which have
5-14 ring atoms wherein 1 to 4 heteroatoms are ring atoms in the
aromatic ring and the remainder of the ring atoms being carbon
atoms. Suitable heteroatoms include oxygen, sulfur, and nitrogen.
Suitable heteroaryl groups include furanyl, thienyl, pyridyl,
pyrrolyl, N-lower alkyl pyrrolyl, pyridyl-N-oxide, pyrimidyl,
pyrazinyl, imidazolyl, indolyl and the like, all optionally
substituted.
[0058] The term "monocyclic aryl" refers to aromatic groups which
have 5-6 ring atoms and includes carbocyclic aryl and heterocyclic
aryl. Suitable aryl groups include phenyl, furanyl, pyridyl, and
thienyl. Aryl groups may be substituted.
[0059] The term "monocyclic heteroaryl" refers to aromatic groups
which have 5-6 ring atoms wherein 1 to 4 heteroatoms are ring atoms
in the aromatic ring and the remainder of the ring atoms being
carbon atoms. Suitable heteroatoms include oxygen, sulfur, and
nitrogen.
[0060] The term "biaryl" represents aryl groups which have 5-14
atoms containing more than one aromatic ring including both fused
ring systems and aryl groups substituted with other aryl groups.
Such groups may be optionally substituted. Suitable biaryl groups
include naphthyl and biphenyl.
[0061] The term "optionally substituted" or "substituted" includes
groups substituted by one to four substituents, independently
selected from lower alkyl, lower aryl, lower aralkyl, lower cyclic
alkyl, lower heterocycloalkyl, hydroxy, lower alkoxy, lower
aryloxy, perhaloalkoxy, aralkoxy, lower heteroaryl, lower
heteroaryloxy, lower heteroarylalkyl, lower heteroaralkoxy, azido,
amino, halogen, lower alkylthio, oxo, lower acylalkyl, lower
carboxy esters, carboxyl, -carboxamido, nitro, lower acyloxy, lower
aminoalkyl, lower alkylaminoaryl, lower alkylaryl, lower
alkylaminoalkyl, lower alkoxyaryl, lower arylamino, lower
aralkylamino, lower alkylsulfonyl, lower-carboxamidoalkylaryl,
lower-carboxamidoaryl, lower hydroxyalkyl, lower haloalkyl, lower
alkylaminoalkylcarboxy-, lower aminocarboxamidoalkyl-, cyano, lower
alkoxyalkyl, lower perhaloalkyl, and lower arylalkyloxyalkyl.
"Substituted aryl" and "substituted heteroaryl" refers to aryl and
heteroaryl groups substituted with 1-6 substituents. These
substituents are selected from the group consisting of lower alkyl,
lower alkoxy, lower perhaloalkyl, halogen, hydroxy, cyano, and
amino.
[0062] The term "-aralkyl" refers to an alkylene group substituted
with an aryl group. Suitable aralkyl groups include benzyl,
picolyl, and the like, and may be optionally substituted. The aryl
portion may have 5-14 ring atoms and the alkyl portion may have up
to and including 10 carbon atoms. "Heteroarylalkyl" refers to an
alkylene group substituted with a heteroaryl group.
[0063] The term "alkylaryl-" refers to an aryl group substituted
with an alkyl group. "Lower alkylaryl-" refers to such groups where
alkyl is lower alkyl. The aryl portion may have 5-14 ring atoms and
the alkyl portion may have up to and including 10 carbon atoms. The
term "lower" referred to herein in connection with organic radicals
or compounds respectively defines such as with up to and including
10, in one aspect up to and including 6, and in another aspect one
to four carbon atoms. Such groups may be straight chain, branched,
or cyclic.
[0064] The term "cyclic alkyl" or "cycloalkyl" refers to alkyl
groups that are cyclic of 3 to 10 carbon atoms, and in one aspect
are 3 to 6 carbon atoms. Suitable cyclic groups include norbornyl
and cyclopropyl. Such groups may be substituted.
[0065] The term "heterocyclic", "heterocyclic alkyl" or
"heterocycloalkyl" refer to cyclic groups of 3 to 10 atoms, and in
one aspect are 3 to 6 atoms, containing at least one heteroatom, in
a further aspect are 1 to 3 heteroatoms. Suitable heteroatoms
include oxygen, sulfur, and nitrogen. Heterocyclic groups may be
attached through a nitrogen or through a carbon atom in the ring.
The heterocyclic alkyl groups include unsaturated cyclic, fused
cyclic and spirocyclic groups. Suitable heterocyclic groups include
pyrrolidinyl, morpholino, morpholinoethyl, and pyridyl.
[0066] The terms "arylamino" (a), and "aralkylamino" (b),
respectively, refer to the group --NRR' wherein respectively, (a) R
is aryl and R' is hydrogen, alkyl, aralkyl, heterocycloalkyl, or
aryl, and (b) R is aralkyl and R' is hydrogen, aralkyl, aryl, alkyl
or heterocycloalkyl.
[0067] The term "acyl" refers to --C(O)R where R is alkyl,
heterocycloalkyl, or aryl. The term "lower acyl" refers to where R
is lower alkyl. The term C.sub.1-C.sub.4 acyl refers to where R is
C.sub.1-C.sub.4.
[0068] The term "carboxy esters" refers to --C(O)OR where R is
alkyl, aryl, aralkyl, cyclic alkyl, or heterocycloalkyl, all
optionally substituted.
[0069] The term "carboxyl" refers to --C(O)OH.
[0070] The term "oxo" refers to .dbd.O in an alkyl or
heterocycloalkyl group.
[0071] The term "amino" refers to --NRR' where R and R' are
independently selected from hydrogen, alkyl, aryl, aralkyl and
heterocycloalkyl, all except H are optionally substituted; and R
and R' can form a cyclic ring system.
[0072] The term "-carboxylamido" refers to --CONR.sub.2 where each
R is independently hydrogen or alkyl.
[0073] The term "-sulphonylamido" or "-sulfonylamido" refers to
--S(.dbd.O).sub.2NR.sub.2 where each R is independently hydrogen or
alkyl.
[0074] The term "halogen" or "halo" refers to --F, --Cl, --Br and
--I.
[0075] The term "alkylaminoalkylcarboxy" refers to the group
alkyl-NR-alk-C(O)--O-- where "alk" is an alkylene group, and R is a
H or lower alkyl.
[0076] The term "sulphonyl" or "sulfonyl" refers to --SO.sub.2R,
where R is H, alkyl, aryl, aralkyl, or heterocycloalkyl.
[0077] The term "sulphonate" or "sulfonate" refers to --SO.sub.2OR,
where R is --H, alkyl, aryl, aralkyl, or heterocycloalkyl.
[0078] The term "alkenyl" refers to unsaturated groups which have 2
to 12 atoms and contain at least one carbon-carbon double bond and
includes straight-chain, branched-chain and cyclic groups. Alkenyl
groups may be optionally substituted. Suitable alkenyl groups
include allyl. "1-Alkenyl" refers to alkenyl groups where the
double bond is between the first and second carbon atom. If the
1-alkenyl group is attached to another group, e.g. it is a W
substituent attached to the cyclic phosphate, it is attached at the
first carbon.
[0079] The term "alkynyl" refers to unsaturated groups which have 2
to 12 atoms and contain at least one carbon-carbon triple bond and
includes straight-chain, branched-chain and cyclic groups. Alkynyl
groups may be optionally substituted. Suitable alkynyl groups
include ethynyl. "1-Alkynyl" refers to alkynyl groups where the
triple bond is between the first and second carbon atom. If the
1-alkynyl group is attached to another group, e.g. it is a W
substituent attached to the cyclic phosphate, it is attached at the
first carbon.
[0080] The term "alkylene" refers to a divalent straight chain,
branched chain or cyclic saturated aliphatic group. In one aspect
the alkylene group contains up to and including 10 atoms. In
another aspect the alkylene chain contains up to and including 6
atoms. In a further aspect the alkylene groups contains up to and
including 4 atoms. The alkylene group can be either straight,
branched or cyclic. The alkylene may be optionally substituted with
1-3 substituents.
[0081] The term "acyloxy" refers to the ester group --O--C(O)R,
where R is H, alkyl, alkenyl, alkynyl, aryl, aralkyl, or
heterocycloalkyl.
[0082] The term "aminoalkyl-" refers to the group NR.sub.2-alk-
wherein "alk" is an alkylene group and R is selected from --H,
alkyl, aryl, aralkyl, and heterocycloalkyl.
[0083] The term "alkylaminoalkyl-" refers to the group
alkyl-NR-alk- wherein each "alk" is an independently selected
alkylene, and R is H or lower alkyl. "Lower alkylaminoalkyl-"
refers to groups where the alkyl and the alkylene group is lower
alkyl and alkylene, respectively.
[0084] The term "arylaminoalkyl-" refers to the group aryl-NR-alk-
wherein "alk" is an alkylene group and R is --H, alkyl, aryl,
aralkyl, or heterocycloalkyl. In "lower arylaminoalkyl-", the
alkylene group is lower alkylene.
[0085] The term "alkylaminoaryl-" refers to the group
alkyl-NR-aryl- wherein "aryl" is a divalent group and R is --H,
alkyl, aralkyl, or heterocycloalkyl. In "lower alkylaminoaryl-",
the alkyl group is lower alkyl.
[0086] The term "alkoxyaryl-" refers to an aryl group substituted
with an alkyloxy group. In "lower alkyloxyaryl-", the alkyl group
is lower alkyl.
[0087] The term "aryloxyalkyl-" refers to an alkyl group
substituted with an aryloxy group.
[0088] The term "aralkyloxyalkyl-" refers to the group
aryl-alk-O-alk- wherein "alk" is an alkylene group. "Lower
aralkyloxyalkyl-" refers to such groups where the alkylene groups
are lower alkylene.
[0089] The term "alkoxy-" or "alkyloxy-" refers to the group
alkyl-O--.
[0090] The term "alkoxyalkyl-" or "alkyloxyalkyl-" refer to the
group alkyl-O-alk- wherein "alk" is an alkylene group. In "lower
alkoxyalkyl-", each alkyl and alkylene is lower alkyl and alkylene,
respectively.
[0091] The terms "alkylthio-" refers to the group alkyl-S--.
[0092] The term "alkylthioalkyl-" refers to the group alkyl-S-alk-
wherein "alk" is an alkylene group. In "lower alkylthioalkyl-" each
alkyl and alkylene is lower alkyl and alkylene, respectively.
[0093] The term "alkoxycarbonyloxy-" refers to
alkyl-O--C(O)--O--.
[0094] The term "aryloxycarbonyloxy-" refers to
aryl-O--C(O)--O--.
[0095] The term "alkylthiocarbonyloxy-" refers to
alkyl-S--C(O)--O--.
[0096] The term "amido" refers to the NR.sub.2 group next to an
acyl or sulfonyl group as in NR.sub.2--C(O)--, RC(O)--NR.sup.1--,
NR.sub.2--S(.dbd.O).sub.2-- and RS(.dbd.O).sub.2--NR.sup.1--, where
R and R.sup.1 include --H, alkyl, aryl, aralkyl, and
heterocycloalkyl.
[0097] The term "carboxamido" refer to NR.sub.2--C(O)-- and
RC(O)--NR.sup.1--, where R and R.sup.1 include --H, alkyl, aryl,
aralkyl, and heterocycloalkyl. The term does not include urea,
--NR--C(O)--NR--.
[0098] The terms "sulphonamido" or "sulfonamido" refer to
NR.sub.2--S(.dbd.O).sub.2-- and RS(.dbd.O).sub.2--NR.sup.1--, where
R and R.sup.1 include --H, alkyl, aryl, aralkyl, and
heterocycloalkyl. The term does not include sulfonylurea,
--NR--S(.dbd.O).sub.2--NR--.
[0099] The term "carboxamidoalkylaryl" and "carboxamidoaryl" refers
to an aryl-alk-NR.sup.1--C(O), and ar-NR.sup.1--C(O)-alk-,
respectively where "ar" is aryl, "alk" is alkylene, R.sup.1 and R
include H, alkyl, aryl, aralkyl, and heterocycloalkyl.
[0100] The term "sulfonamidoalkylaryl" and "sulfonamidoaryl" refers
to an aryl-alk-NR.sup.1--S(.dbd.O).sub.2--, and
ar-NR.sup.1--S(.dbd.O).sub.2--, respectively where "ar" is aryl,
"alk" is alkylene, R.sup.1 and R include --H, alkyl, aryl, aralkyl,
and heterocycloalkyl.
[0101] The term "hydroxyalkyl" refers to an alkyl group substituted
with one --OH.
[0102] The term "haloalkyl" refers to an alkyl group substituted
with one halogen.
[0103] The term "cyano" refers to --C.ident.N.
[0104] The term "nitro" refers to --NO.sub.2.
[0105] The term "acylalkyl" refers to an alkyl-C(O)-alk-, where
"alk" is alkylene.
[0106] The term "aminocarboxamidoalkyl-" refers to the group
NR.sub.2--C(O)--N(R)-alk- wherein R is an alkyl group or H and
"alk" is an alkylene group. "Lower aminocarboxamidoalkyl-" refers
to such groups wherein "alk" is lower alkylene.
[0107] The term "heteroarylalkyl" refers to an alkylene group
substituted with a heteroaryl group.
[0108] The term "perhalo" refers to groups wherein every C--H bond
has been replaced with a C-halo bond on an aliphatic or aryl group.
Suitable perhaloalkyl groups include --CF.sub.3 and
--CFCl.sub.2.
[0109] The term "heterocyclic base B" refers to ##STR8##
[0110] wherein:
[0111] R.sup.14 is independently selected from the group consisting
of H and NH.sub.2; and X is selected from the group consisting of
NH.sub.2, NHCH.sub.3, N(CH.sub.3).sub.2, OCH.sub.3, SCH.sub.3, OH,
and SH.
[0112] The phrase "therapeutically effective amount" means an
amount of a compound or a combination of compounds that
ameliorates, attenuates or eliminates one or more of the symptoms
of a particular disease or condition or prevents, modifies, or
delays the onset of one or more of the symptoms of a particular
disease or condition.
[0113] The term "pharmaceutically acceptable salt" includes salts
of compounds of Formula I and its prodrugs derived from the
combination of a compound of this invention and an organic or
inorganic acid or base. Suitable acids include acetic acid, adipic
acid, benzenesulfonic acid,
(+)-7,7-dimethyl-2-oxobicyclo[2.2.1]heptane-1-methanesulfonic acid,
citric acid, 1,2-ethanedisulfonic acid, dodecyl sulfonic acid,
fumaric acid, glucoheptonic acid, gluconic acid, glucuronic acid,
hippuric acid, hydrochloride hemiethanolic acid, HBr, HCl, HI,
2-hydroxyethanesulfonic acid, lactic acid, lactobionic acid, maleic
acid, methanesulfonic acid, methylbromide acid, methyl sulfuric
acid, 2-naphthalenesulfonic acid, nitric acid, oleic acid,
4,4'-methylenebis[3-hydroxy-2-naphthalenecarboxylic acid],
phosphoric acid, polygalacturonic acid, stearic acid, succinic
acid, sulfuric acid, sulfosalicylic acid, tannic acid, tartaric
acid, terphthalic acid, and p-toluenesulfonic acid.
[0114] The term "naturally-occurring L-amino acid" refers to those
amino acids routinely found as components of proteinaceous
molecules in nature, including alanine, valine, leucine,
isoleucine, proline, phenylalanine, tryptophan, methionine,
glycine, serine, threonine, cysteine, tyrosine, asparagine,
glutamine, aspartic acid, glutamic acid, lysine, arginine and
histidine. In one aspect, this term is intended to encompass
L-amino acids having only the amine and carboxylic acid as charged
functional groups, i.e., alanine, valine, leucine, isoleucine,
proline, phenylalanine, tryptophan, methionine, glycine, serine,
threonine, cysteine and tyrosine. In another aspect they are
alanine, valine, leucine, isoleucine, proline, phenylalanine, and
glycine. In a further aspect, it is valine.
[0115] The term "patient" refers to an animal being treated
including a mammal, such as a dog, a cat, a cow, a horse, a sheep,
and a human. Another aspect includes a mammal, both male and
female.
[0116] The term "prodrug" as used herein refers to any compound
that when administered to a biological system generates a
biologically active compound as a result of spontaneous chemical
reaction(s), enzyme catalyzed chemical reaction(s), and/or
metabolic chemical reaction(s), or a combination of each. Standard
prodrugs are formed using groups attached to functionality, e.g.
HO--, HS--, HOOC--, R.sub.2N--, associated with the drug, that
cleave in vivo. Standard prodrugs include but are not limited to
carboxylate esters where the group is alkyl, aryl, aralkyl,
acyloxyalkyl, alkoxycarbonyloxyalkyl as well as esters of hydroxyl,
thiol and amines where the group attached is an acyl group, an
alkoxycarbonyl, aminocarbonyl, phosphate or sulfate. The groups
illustrated are exemplary, not exhaustive, and one skilled in the
art could prepare other known varieties of prodrugs. Such prodrugs
of the compounds of Formula I fall within this scope. Prodrugs must
undergo some form of a chemical transformation to produce the
compound that is biologically active or is a precursor of the
biologically active compound. In some cases, the prodrug is
biologically active, usually less than the drug itself, and serves
to improve drug efficacy or safety through improved oral
bioavailability, pharmacodynamic half-life, etc. Prodrug forms of
compounds may be utilized, for example, to improve bioavailability,
improve subject acceptability such as by masking or reducing
unpleasant characteristics such as bitter taste or gastrointestinal
irritability, alter solubility such as for intravenous use, provide
for prolonged or sustained release or delivery, improve ease of
formulation, or provide site-specific delivery of the compound.
Prodrugs are described in The Organic Chemistry of Drug Design and
Drug Action, by Richard B. Silverman, Academic Press, San Diego,
1992. Chapter 8: "Prodrugs and Drug delivery Systems" pp. 352-401;
Design of Prodrugs, edited by H. Bundgaard, Elsevier Science,
Amsterdam, 1985; Design of Biopharmaceutical Properties through
Prodrugs and Analogs, Ed. by E. B. Roche, American Pharmaceutical
Association, Washington, 1977; and Drug Delivery Systems, ed. by R.
L. Juliano, Oxford Univ. Press, Oxford, 1980.
[0117] The term "prodrug" herein also includes but is not limited
to esterase cleavable prodrugs of the 2' and 3'-hydroxy groups of
compounds of Formula I (Anastasi et al., Curr. Med. Chem., 2003,
10, 1825). Standard groups include acyl and alkoxycarbonyl groups,
and esters of natural L-amino acid derivatives (Perry, et al.,
Drugs, 1996, 52, 754). Also included is a cyclic carbonate
derivative formed by carbonylation of the 2' and 3'-hydroxy groups,
which upon activation by esterase activity in vivo results in
compounds of Formula I.
[0118] In the case of bases, "prodrugs" are preferred at the
6-position of purine analogs. Such substitution may include H,
halogen, amino, acetoxy or azido groups. Hydrogen substituted
prodrugs at the 6-position of guanosine analogs undergo oxidation
in vivo by aldehyde oxidase or xanthine oxidase to give the
required functionality (Rashidi et al., Drug Metab. Dispos. 1997,
25, 805). While esterases unmask acetoxy groups, amine and halogen
substituents are known to be substrates for deaminases. 6-Azido
substituted compounds are also known to give the corresponding
amino derivatives by the action of reductases (Koudriakova, et al.,
J. Med Chem., 1996, 39, 4676).
[0119] The structure ##STR9## [0120] has a plane of symmetry
running through the phosphorus-oxygen double bond when V=W and V
and W are either both pointing up or both pointing down.
[0121] The term "cyclic phosphate ester of 1,3-propanediol",
"cyclic phosphate diester of 1,3-propanediol", "2 oxo
2.lamda..sup.5[1,3,2]dioxaphosphorinane",
"2-oxo-[1,3,2]-dioxaphosphorinane", or "dioxaphosphorinane" refers
to the following: ##STR10##
[0122] The phrase "together V and Z are connected via an additional
3-5 atoms to form a cyclic group, optionally containing one
heteroatom, that is fused to an aryl group attached at the beta and
gamma position to the O attached to the phosphorus" includes the
following: ##STR11##
[0123] As shown above together V and Z are connected via 4
additional atoms.
[0124] The phrase "together W and W' are connected via an
additional 2-5 atoms to form a cyclic group, optionally containing
0-2 heteroatoms, and V must be aryl, substituted aryl, heteroaryl,
or substituted heteroaryl" includes the following: ##STR12##
[0125] As shown above together W and W' are connected via an
additional 2 atoms.
[0126] The structure above has V=aryl, and a spiro-fused
cyclopropyl group for W and W'.
[0127] The term "cyclic phosphate" refers to ##STR13##
[0128] The carbon attached to V must have a C--H bond. The carbon
attached to Z must also have a C--H bond.
[0129] The term "cis" stereochemistry refers to the spatial
relationship of the V group and the substituent attached to the
phosphorus atom via an exocyclic single bond on the six membered
2-oxo-phosphorinane ring. The structures A and B below show two
possible cis-isomers of 2- and 4- substituted 2-oxo-phosphorinane.
Structure A shows cis-isomer of (2S, 4R)-configuration whereas
structure B shows cis-isomer of (2R, 4S)-configuration.
##STR14##
[0130] The term "trans" stereochemistry refers to the spatial
relationship of the V group and the substituent attached to the
phosphorus atom via an exocyclic single bond on the six membered
2-oxo-phosphorinane ring. The structures C and D below show two
possible trans-isomers of 2- and 4-substituted 2-oxo-phosphorinane.
Structure C shows trans-isomer of (2S, 4S)-- configuration whereas
structure D shows trans-isomer of (2R, 4R)-configuration.
##STR15##
[0131] The term "percent enantiomeric excess (% ee)" refers to
optical purity. It is obtained by using the following formula: [ R
] - [ S ] [ R ] + [ S ] .times. 100 = % .times. .times. R - %
.times. .times. S ##EQU1##
[0132] where [R] is the amount of the R isomer and [S] is the
amount of the S isomer. This formula provides the % ee when R is
the dominant isomer.
[0133] The term "enantioenriched" or "enantiomerically enriched"
refers to a sample of a chiral compound that consists of more of
one enantiomer than the other. The extent to which a sample is
enantiomerically enriched is quantitated by the enantiomeric ratio
or the enantiomeric excess.
[0134] The term "liver" refers to liver organ.
[0135] The term "enhancing" refers to increasing or improving a
specific property.
[0136] The term "liver specificity" refers to the ratio: [ drug
.times. .times. or .times. .times. a .times. .times. drug .times.
.times. metabolite .times. .times. in .times. .times. liver .times.
.times. tissue ] [ drug .times. .times. or .times. .times. a
.times. .times. drug .times. .times. metabolite .times. .times. in
.times. .times. blood .times. .times. or .times. .times. another
.times. .times. tissue ] ##EQU2## as measured in animals treated
with the drug or a prodrug. The ratio can be determined by
measuring tissue levels at a specific time or may represent an AUC
based on values measured at three or more time points.
[0137] The term "increased or enhanced liver specificity" refers to
an increase in the liver specificity ratio in animals treated with
the prodrug relative to animals treated with the parent drug.
[0138] The term "enhanced oral bioavailability" refers to an
increase of at least 50% of the absorption of the dose of the
parent drug. In an additional aspect the increase in oral
bioavailability of the prodrug (compared to the parent drug) is at
least 100%, that is a doubling of the absorption. Measurement of
oral bioavailability usually refers to measurements of the prodrug,
drug, or drug metabolite in blood, plasma, tissues, or urine
following oral administration compared to measurements following
parenteral administration.
[0139] The term "therapeutic index" refers to the ratio of the dose
of a drug or prodrug that produces a therapeutically beneficial
response relative to the dose that produces an undesired response
such as death, an elevation of markers that are indicative of
toxicity, and/or pharmacological side effects.
[0140] The term "sustained delivery" refers to an increase in the
period in which there is a prolongation of
therapeutically-effective drug levels due to the presence of the
prodrug.
[0141] The term "bypassing drug resistance" refers to the loss or
partial loss of therapeutic effectiveness of a drug (drug
resistance) due to changes in the biochemical pathways and cellular
activities important for producing and maintaining the biological
activity of the drug and the ability of an agent to bypass this
resistance through the use of alternative pathways or the failure
of the agent to induce changes that tend to resistance.
[0142] The terms "treating" or "treatment" of a disease includes
inhibiting the disease (slowing or arresting its development),
providing relief from the symptoms or side-effects of the disease
(including palliative treatment), and relieving the disease
(causing regression of the disease).
DETAILED DESCRIPTION
[0143] The present invention relates to compounds of Formula I,
stereoisomers, pharmaceutically acceptable salts or prodrugs
thereof or pharmaceutically acceptable salts of the prodrugs as
represented by Formula I: ##STR16##
[0144] wherein:
[0145] B is selected from the group consisting of ##STR17##
[0146] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl;
[0147] W and W' are independently selected from the group
consisting of --R.sup.2, optionally substituted monocyclic aryl,
and optionally substituted monocyclic heteroaryl;
[0148] Z is selected from the group consisting of halogen, --CN,
--COR.sup.5, --CONR.sup.4.sub.2, --CO.sub.2R.sup.5,
--SO.sub.2R.sup.5, --SO.sub.2NR.sup.4.sub.2, --OR.sup.4,
--SR.sup.4, --R.sup.4, --NR.sup.4.sub.2, --OCOR.sup.5,
--OCO.sub.2R.sup.5, --SCOR.sup.5, --SCO.sub.2R.sup.5,
--NHCOR.sup.4, --NHCO.sub.2R.sup.5, --(CH.sub.2).sub.p--OR.sup.6,
and --(CH.sub.2).sub.p--SR.sup.6; or
[0149] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; or
[0150] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0151] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group, optionally containing 0-2 heteroatoms;
[0152] R.sup.2 is selected from the group consisting of R.sup.3 and
hydrogen;
[0153] R.sup.3 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0154] R.sup.4 is selected from the group consisting of R.sup.3 and
hydrogen;
[0155] R.sup.5 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0156] R.sup.6 is selected from the group consisting of hydrogen,
and lower acyl;
[0157] R.sup.12 is selected from the group consisting of hydrogen,
and lower acyl; and
[0158] p is an integer 2 or 3;
[0159] or pharmaceutically acceptable prodrugs or salts
thereof.
[0160] In one aspect, the invention comprises compounds of Formula
I: ##STR18##
[0161] wherein:
[0162] B is ##STR19##
[0163] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl;
[0164] W and W' are independently selected from the group
consisting of --R.sup.2, optionally substituted monocyclic aryl,
and optionally substituted monocyclic heteroaryl;
[0165] Z is selected from the group consisting of halogen, --CN,
--COR.sup.5, --CONR.sup.4.sub.2, --CO.sub.2R.sup.5,
--SO.sub.2R.sup.5, --SO.sub.2NR.sup.4.sub.2, --OR.sup.4,
--SR.sup.4, --R.sup.4, --NR.sup.4.sub.2, --OCOR.sup.5,
--OCO.sub.2R.sup.5, --SCOR.sup.5, --SCO.sub.2R.sup.5,
--NHCOR.sup.4, --NHCO.sub.2R.sup.5, --(CH.sub.2).sub.p--OR.sup.6,
and --(CH.sub.2).sub.p--SR.sup.6; or
[0166] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; or
[0167] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0168] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group, optionally containing 0-2 heteroatoms;
[0169] R.sup.2 is selected from the group consisting of R.sup.3 and
hydrogen;
[0170] R.sup.3 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0171] R.sup.4 is selected from the group consisting of R.sup.5 and
hydrogen;
[0172] R.sup.5 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0173] R.sup.6 is selected from the group consisting of hydrogen,
and lower acyl;
[0174] R.sup.12 is selected from the group consisting of hydrogen,
and lower acyl; and
[0175] p is an integer 2 or 3;
or pharmaceutically acceptable prodrugs or salts thereof.
[0176] In another aspect, the invention comprises compounds of
Formula I: ##STR20##
[0177] wherein:
[0178] B is ##STR21##
[0179] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl;
[0180] W and W' are independently selected from the group
consisting of --R.sup.2, optionally substituted monocyclic aryl,
and optionally substituted monocyclic heteroaryl;
[0181] Z is selected from the group consisting of halogen, --CN,
--COR.sup.5, --CONR.sup.4.sub.2, --CO.sub.2R.sup.5,
--SO.sub.2R.sup.5, --SO.sub.2NR.sup.4.sub.2, --OR.sup.4,
--SR.sup.4, --R.sup.4, --NR.sup.4.sub.2, --OCOR.sup.5,
--OCO.sub.2R.sup.5, --SCOR.sup.5, --SCO.sub.2R.sup.5,
--NHCOR.sup.4, --NHCO.sub.2R.sup.5, --(CH.sub.2).sub.p--OR.sup.6,
and --(CH.sub.2).sub.p--SR.sup.6; or
[0182] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; or
[0183] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0184] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group, optionally containing 0-2 heteroatoms;
[0185] R.sup.2 is selected from the group consisting of R.sup.3 and
hydrogen;
[0186] R.sup.3 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0187] R.sup.4 is selected from the group consisting of R.sup.5 and
hydrogen;
[0188] R.sup.5 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0189] R.sup.6 is selected from the group consisting of hydrogen,
and lower acyl;
[0190] R.sup.12 is selected from the group consisting of hydrogen,
and lower acyl; and
[0191] p is an integer 2 or 3;
or pharmaceutically acceptable prodrugs or salts thereof.
[0192] In another aspect, the invention comprises compounds of
Formula I: ##STR22##
[0193] wherein:
[0194] B is ##STR23##
[0195] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl;
[0196] W and W' are independently selected from the group
consisting of --R.sup.2, optionally substituted monocyclic aryl,
and optionally substituted monocyclic heteroaryl;
[0197] Z is selected from the group consisting of halogen, --CN,
--COR.sup.5, --CONR.sup.4.sub.2, --CO.sub.2R.sup.5,
--SO.sub.2R.sup.5, --SO.sub.2NR.sup.4.sub.2, --OR.sup.4,
--SR.sup.4, --R.sup.4, --NR.sup.4.sub.2, --OCOR.sup.5,
--OCO.sub.2R.sup.5, --SCOR.sup.5, --SCO.sub.2R.sup.5,
--NHCOR.sup.4, --NHCO.sub.2R.sup.5, --(CH.sub.2).sub.p--OR.sup.6,
and --(CH.sub.2).sub.p--SR.sup.6; or
[0198] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; or
[0199] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0200] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group, optionally containing 0-2 heteroatoms;
[0201] R.sup.2 is selected from the group consisting of R.sup.3 and
hydrogen;
[0202] R.sup.3 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0203] R.sup.4 is selected from the group consisting of R.sup.5 and
hydrogen;
[0204] R.sup.5 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0205] R.sup.6 is selected from the group consisting of hydrogen,
and lower acyl;
[0206] R.sup.12 is selected from the group consisting of hydrogen,
and lower acyl; and
[0207] p is an integer 2 or 3;
[0208] or pharmaceutically acceptable prodrugs or salts
thereof.
[0209] In yet another aspect, the invention comprises compounds of
Formula I: ##STR24##
[0210] wherein:
[0211] B is ##STR25##
[0212] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl;
[0213] W and W' are independently selected from the group
consisting of --R.sup.2, optionally substituted monocyclic aryl,
and optionally substituted monocyclic heteroaryl;
[0214] Z is selected from the group consisting of halogen, --CN,
--COR.sup.5, --CONR.sup.4.sub.2, --CO.sub.2R.sup.5,
--SO.sub.2R.sup.5, --SO.sub.2NR.sup.4.sub.2, --OR.sup.4,
--SR.sup.4, --R.sup.4, --NR.sup.4.sub.2, --OCOR.sup.5,
--OCO.sub.2R.sup.5, --SCOR.sup.5, --SCO.sub.2R.sup.5,
--NHCOR.sup.4, --NHCO.sub.2R.sup.5, --(CH.sub.2).sub.p--OR.sup.6,
and --(CH.sub.2).sub.p--SR.sup.6; or
[0215] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; or
[0216] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0217] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group, optionally containing 0-2 heteroatoms;
[0218] R.sup.2 is selected from the group consisting of R.sup.3 and
hydrogen;
[0219] R.sup.3 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0220] R.sup.4 is selected from the group consisting of R.sup.5 and
hydrogen;
[0221] R.sup.5 is selected from the group consisting of alkyl,
aryl, heterocycloalkyl, and aralkyl;
[0222] R.sup.6 is selected from the group consisting of hydrogen,
and lower acyl;
[0223] R.sup.12 is selected from the group consisting of hydrogen,
and lower acyl; and
[0224] p is an integer 2 or 3;
[0225] or pharmaceutically acceptable prodrugs or salts
thereof.
[0226] In one aspect, V is selected from the group consisting of
phenyl, substituted phenyl with 1-3 substituents independently
selected from the group consisting of halogen, C.sub.1-C.sub.6
alkyl, --CF.sub.3, --OR.sup.3, --OR.sup.12, --COR.sup.3,
--CO.sub.2R.sup.3, --NR.sup.3.sub.2, --NR.sup.12.sub.2,
--CO.sub.2NR.sub.2.sup.2, --SR.sup.3, --SO.sub.2R.sup.3,
--SO.sub.2NR.sub.2.sup.2 and --CN, monocyclic heteroaryl, and
substituted monocyclic heteroaryl with 1-2 substituents
independently selected from the group consisting of halogen,
C.sub.1-C.sub.6 alkyl, --CF.sub.3, --OR.sup.3, --OR.sup.12,
--COR.sup.3, --CO.sub.2R.sup.3, --NR.sup.3.sub.2, NR.sup.12.sub.2,
--CO.sub.2NR.sub.2.sup.2, --SO.sub.2R.sup.3,
--SO.sub.2NR.sub.2.sup.2 and --CN, and wherein said monocyclic
heteroaryl and substituted monocyclic heteroaryl has 1-2
heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that
[0227] a) when there are two heteroatoms and one is O, then the
other can not be O or S, and
[0228] b) when there are two heteroatoms and one is S, then the
other can not be O or S; or
[0229] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; and R.sup.3 is C.sub.1-C.sub.6
alkyl.
[0230] In another aspect, V is selected from the group consisting
of phenyl, substituted phenyl with 1-3 substituents independently
selected from the group consisting of --Cl, --Br, --F,
C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN, monocyclic heteroaryl,
and substituted monocyclic heteroaryl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN and wherein said
monocyclic heteroaryl and substituted monocyclic heteroaryl has 1-2
heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that
[0231] a) when there are two heteroatoms and one is O, then the
other can not be O or S, and
[0232] b) when there are two heteroatoms and one is S, then the
other can not be O or S; or
[0233] together V and Z are connected via an additional 4 atoms to
form a 6-membered ring that is fused to a phenyl or substituted
phenyl at the beta and gamma position to the O attached to the
phosphorus.
[0234] In yet another aspect, V is selected from the group
consisting of phenyl; substituted phenyl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; pyridyl; substituted
pyridyl with 1 substituent independently selected from the group
consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3; furanyl; substituted furanyl with 1 substituent
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; thienyl; and
substituted thienyl with 1 substituent independently selected from
the group consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3.
[0235] In a further aspect, V is selected from the group consisting
of phenyl, 3-chlorophenyl, 3-bromophenyl, 2-bromophenyl,
3,5-dichlorophenyl, 3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl,
and 4-pyridyl. In another aspect, V is selected from the group
consisting of 3-chlorophenyl, 3-bromophenyl, 2-bromophenyl,
3,5-dichlorophenyl, 3-pyridyl, and 4-pyridyl.
[0236] In another aspect, V is selected from the group consisting
of phenyl, substituted phenyl with 1-3 substituents independently
selected from the group consisting of --Cl, --Br, --F,
C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OH, --OMe,
--NH.sub.2, --NMe.sub.2, --OEt, --COOH, --CO.sub.2t-butyl,
--CO.sub.2NH.sub.2, --SMe, --SO.sub.2Me, --SO.sub.2NH.sub.2 and
--CN; monocyclic heteroaryl, and substituted monocyclic heteroaryl
with 1-2 substituents independently selected from the group
consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, --CF.sub.3,
--COCH.sub.3, --OH, --OMe, --NH.sub.2, --NMe.sub.2, --OEt, --COOH,
--CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe, --SO.sub.2Me,
--SO.sub.2NH.sub.2 and --CN; and wherein said monocyclic heteroaryl
and substituted monocyclic heteroaryl has 1-2 heteroatoms that are
independently selected from the group consisting of N, O, and S
with the provisos that
[0237] a) when there are two heteroatoms and one is O, then the
other can not be O or S, and
[0238] b) when there are two heteroatoms and one is S, then the
other can not be O or S; or
[0239] together V and Z are connected via an additional 4 atoms to
form a 6-membered ring that is fused to a phenyl or substituted
phenyl at the beta and gamma position to the O attached to the
phosphorus.
[0240] In one aspect, Z is selected from the group consisting of
--H, --OMe, --OEt, phenyl, C.sub.1-C.sub.3 alkyl, --NR.sup.4.sub.2,
--SR.sup.4, --(CH.sub.2).sub.p--OR.sup.6,
--(CH.sub.2).sub.p--SR.sup.6 and --OCOR.sup.5; R.sup.4 is
C.sub.1-C.sub.4 alkyl; R.sup.5 is selected from the group
consisting of C.sub.1-C.sub.4 alkyl, monocyclic aryl, and
monocyclic aralkyl; and R.sup.6 is C.sub.1-C.sub.4 acyl. In a
further aspect, Z is selected from the group consisting of --H,
--OMe, --OEt, and phenyl.
[0241] In an additional aspect, W and W' are independently selected
from the group consisting of --H, C.sub.1-C.sub.6 alkyl, and
phenyl; or together W and W' are connected via an additional 2-5
atoms to form a cyclic group. In yet another aspect, W and W' are
independently selected from the group consisting of --H, methyl,
and V, or W and W' are each methyl, with the proviso that when W is
V, then W' is H.
[0242] In one aspect, V is selected from the group consisting of
optionally substituted monocyclic aryl and optionally substituted
monocyclic heteroaryl;
[0243] W and W' are independently selected from the group
consisting of --H, methyl, and V, or W and W' are each methyl, with
the proviso that when W is V, then W' is H;
[0244] Z is selected from the group consisting of --H, --OMe,
--OEt, phenyl, C.sub.1-C.sub.3 alkyl, --NR.sup.4.sub.2, --SR.sup.4,
--(CH.sub.2).sub.p--OR.sup.6, --(CH.sub.2).sub.p--SR.sup.6 and
--OCOR.sup.5; or
[0245] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; or
[0246] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0247] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group; and
[0248] R.sup.4 is C.sub.1-C.sub.4 alkyl; R.sup.5 is selected from
the group consisting of C.sub.1-C.sub.4 alkyl, monocyclic aryl, and
monocyclic aralkyl; and R.sup.6 is C.sub.1-C.sub.4 acyl.
[0249] In another aspect, V is selected from the group consisting
of phenyl, substituted phenyl with 1-3 substituents independently
selected from the group consisting of halogen, C.sub.1-C.sub.6
alkyl, --CF.sub.3, --OR.sup.3, --OR.sup.12, --COR.sup.3,
--CO.sub.2R.sup.3, --NR.sup.3.sub.2, --NR.sup.12.sub.2,
--CO.sub.2NR.sub.2.sup.2, --SR.sup.3, --SO.sub.2R.sup.3,
--SO.sub.2NR.sub.2.sup.2 and --CN, monocyclic heteroaryl, and
substituted monocyclic heteroaryl with 1-2 substituents
independently selected from the group consisting of halogen,
C.sub.1-C.sub.6 alkyl, --CF.sub.3, --OR.sup.3, --OR.sup.12,
--COR.sup.3, --CO.sub.2R.sup.3, --NR.sup.3.sub.2,
--NR.sup.12.sub.2, --CO.sub.2NR.sub.2.sup.2, --SR.sup.3,
--SO.sub.2R.sup.3, --SO.sub.2NR.sub.2.sup.2 and --CN, and wherein
said monocyclic heteroaryl and substituted monocyclic heteroaryl
has 1-2 heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that
[0250] a) when there are two heteroatoms and one is O, then the
other can not be O or S, and
[0251] b) when there are two heteroatoms and one is S, then the
other can not be O or S; or
[0252] W and W' are independently selected from the group
consisting of --H, methyl, and V, or W and W' are each methyl, with
the proviso that when W is V, then W' is H;
[0253] Z is selected from the group consisting of --H, --OMe,
--OEt, phenyl, C.sub.1-C.sub.3 alkyl, --NR.sup.4.sub.2, --SR.sup.4,
--(CH.sub.2).sub.p--OR.sup.6, --(CH.sub.2).sub.p--SR.sup.6 and
--OCOR.sup.5; or
[0254] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; or
[0255] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0256] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group; and
[0257] R.sup.3 is C.sub.1-C.sub.6 alkyl; R.sup.4 is C.sub.1-C.sub.4
alkyl; R.sup.5 is selected from the group consisting of
C.sub.1-C.sub.4 alkyl, monocyclic aryl, and monocyclic aralkyl; and
R.sup.6 is C.sub.1-C.sub.4 acyl.
[0258] In a further aspect, V is selected from the group consisting
of phenyl, substituted phenyl with 1-3 substituents independently
selected from the group consisting of --Cl, --Br, --F,
C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN, monocyclic heteroaryl,
and substituted monocyclic heteroaryl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN, and wherein said
monocyclic heteroaryl and substituted monocyclic heteroaryl has 1-2
heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that
[0259] a) when there are two heteroatoms and one is O, then the
other can not be O or S; and
[0260] b) when there are two heteroatoms and one is S, then the
other can not be O or S; or
[0261] W and W' are independently selected from the group
consisting of --H, methyl, and V, or W and W' are each methyl, with
the proviso that when W is V, then W' is H;
[0262] Z is selected from the group consisting of --H, --OMe,
--OEt, phenyl, C.sub.1-C.sub.3 alkyl, --NR.sup.4.sub.2, --SR.sup.4,
--(CH.sub.2).sub.p--OR.sup.6, --(CH.sub.2).sub.p--SR.sup.6 and
--OCOR.sup.5; or
[0263] together V and Z are connected via an additional 4 atoms to
form a 6-membered ring that is fused to a phenyl or substituted
phenyl at the beta and gamma position to the O attached to the
phosphorus; or
[0264] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0265] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group; and
[0266] R.sup.4 is C.sub.1-C.sub.4 alkyl; R.sup.5 is selected from
the group consisting of C.sub.1-C.sub.4 alkyl, monocyclic aryl, and
monocyclic aralkyl; and R.sup.6 is C.sub.1-C.sub.4 acyl.
[0267] In yet another aspect, V is selected from the group
consisting of phenyl; substituted phenyl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; pyridyl; substituted
pyridyl with 1 substituent independently selected from the group
consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3; furanyl; substituted furanyl with 1 substituent
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; thienyl; and
substituted thienyl with 1 substituent independently selected from
the group consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3;
[0268] W and W' are independently selected from the group
consisting of --H, methyl, and V, or W and W' are each methyl, with
the proviso that when W is V, then W' is H;
[0269] Z is selected from the group consisting of --H, --OMe,
--OEt, phenyl, C.sub.1-C.sub.3 alkyl, --NR.sup.4.sub.2, --SR.sup.4,
--(CH.sub.2).sub.p--OR.sup.6, --(CH.sub.2).sub.p--SR.sup.6 and
--OCOR.sup.5; or
[0270] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0271] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group; and
R.sup.4 is C.sub.1-C.sub.4 alkyl; R.sup.5 is selected from the
group consisting of C.sub.1-C.sub.4 alkyl, monocyclic aryl, and
monocyclic aralkyl; and R.sup.6 is C.sub.1-C.sub.4 acyl.
[0272] In a further aspect, V is selected from the group consisting
of phenyl, 3-chlorophenyl, 3-bromophenyl, 2-bromophenyl,
3,5-dichlorophenyl, 3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl,
and 4-pyridyl; and
[0273] Z is selected from the group consisting of --H, OMe, OEt,
and phenyl; and
[0274] W and W' are independently selected from the group
consisting of --H and phenyl, or W and W' are each methyl.
[0275] In one aspect, Z, W, and W' are each --H. In another aspect,
V and W are the same and each is selected from the group consisting
of optionally substituted monocyclic aryl and optionally
substituted monocyclic heteroaryl.
[0276] In another aspect, B is ##STR26##
[0277] V is selected from the group consisting of 3-chlorophenyl,
3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl, and 4-pyridyl;
and Z, W, and W' are each --H.
[0278] In yet another aspect, B is ##STR27##
[0279] V is selected from the group consisting of 3-chlorophenyl,
3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl, and 4-pyridyl;
and Z, W, and W' are each --H.
[0280] In a further aspect, B is ##STR28##
[0281] V is selected from the group consisting of 3-chlorophenyl,
3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl, and 4-pyridyl;
and Z, W, and W' are each --H.
[0282] In an additional aspect, B is ##STR29##
[0283] V is selected from the group consisting of 3-chlorophenyl,
3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl, and 4-pyridyl;
and Z, W, and W' are each --H.
[0284] A further aspect of this invention includes compounds of
Formula V: ##STR30##
[0285] wherein:
[0286] V and the 5'oxymethylene group of the ribose sugar moiety
are cis to one another
[0287] B is selected from the group consisting of ##STR31##
[0288] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl; or pharmaceutically acceptable prodrugs or salts
thereof.
[0289] In a further aspect, this invention includes compounds of
Formula V: ##STR32##
[0290] wherein:
[0291] V and the 5'oxymethylene group of the ribose sugar moiety
are cis to one another;
[0292] B is ##STR33##
[0293] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl; or pharmaceutically acceptable prodrugs or salts
thereof.
[0294] In an additional aspect, V is selected from the group
consisting of phenyl, substituted phenyl with 1-3 substituents
independently selected from the group consisting of halogen,
C.sub.1-C.sub.6 alkyl, --CF.sub.3, --OR.sup.3, --OR.sup.12,
--COR.sup.3, --CO.sub.2R.sup.3, --NR.sup.3.sub.2,
--NR.sup.12.sub.2, --CO.sub.2NR.sub.2.sup.2, --SR.sup.3,
--SO.sub.2R.sup.3, --SO.sub.2NR.sub.2.sup.2 and --CN, monocyclic
heteroaryl, and substituted monocyclic heteroaryl with 1-2
substituents independently selected from the group consisting of
halogen, C.sub.1-C.sub.6 alkyl, --CF.sub.3, --OR.sup.3,
--OR.sup.12, --COR.sup.3, --CO.sub.2R.sup.3, --NR.sup.3.sub.2,
--NR.sup.12.sub.2, --CO.sub.2NR.sub.2.sup.2, --SR.sup.3,
--SO.sub.2R.sup.3, --SO.sub.2NR.sub.2.sup.2 and --CN, and wherein
said monocyclic heteroaryl and substituted monocyclic heteroaryl
has 1-2 heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that
[0295] a) when there are two heteroatoms and one is O, then the
other can not be O or S, and
[0296] b) when there are two heteroatoms and one is S, then the
other can not be O or S; and R.sup.3 is C.sub.1-C.sub.6 alkyl.
[0297] In a further aspect, V is selected from the group consisting
of phenyl, substituted phenyl with 1-3 substituents independently
selected from the group consisting of --Cl, --Br, --F,
C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN, monocyclic heteroaryl,
and substituted monocyclic heteroaryl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN and wherein said
monocyclic heteroaryl and substituted monocyclic heteroaryl has 1-2
heteroatoms that are independently selected from the group
consisting of N, O, and S with the provisos that
[0298] a) when there are two heteroatoms and one is O, then the
other can not be O or S, and
[0299] b) when there are two heteroatoms and one is S, then the
other can not be O or S; or
[0300] together V and Z are connected via an additional 4 atoms to
form a 6-membered ring that is fused to a phenyl or substituted
phenyl at the beta and gamma position to the O attached to the
phosphorus.
[0301] In an additional aspect, V is selected from the group
consisting of phenyl; substituted phenyl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; pyridyl; substituted
pyridyl with 1 substituent independently selected from the group
consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3; furanyl; substituted furanyl with 1 substituent
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, and --CF.sub.3; thienyl; and
substituted thienyl with 1 substituent independently selected from
the group consisting of --Cl, --Br, --F, C.sub.1-C.sub.3 alkyl, and
--CF.sub.3.
[0302] In yet another aspect, V is selected from the group
consisting of phenyl, 3-chlorophenyl, 3-bromophenyl, 2-bromophenyl,
3,5-dichlorophenyl, 3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl,
and 4-pyridyl. In another aspect, V is selected from the group
consisting of 3-chlorophenyl, 3-bromophenyl, 2-bromophenyl,
3,5-dichlorophenyl, 3-pyridyl, and 4-pyridyl.
[0303] In a further aspect, this invention includes compounds of
Formula II: ##STR34##
[0304] wherein:
[0305] B is selected from the group consisting of: ##STR35##
[0306] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl;
[0307] W and W' are independently selected from the group
consisting of --H, methyl, and V, or W and W' are each methyl, with
the proviso that when W is V, then W' is H;
[0308] Z is selected from the group consisting of --H, --OMe,
--OEt, phenyl, C.sub.1-C.sub.3 alkyl, --NR.sup.4.sub.2, --SR.sup.4,
--(CH.sub.2).sub.p--OR.sup.6, --(CH.sub.2).sub.p--SR.sup.6 and
--OCOR.sup.5; or
[0309] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; or
[0310] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0311] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group;
[0312] R.sup.4 is C.sub.1-C.sub.4 alkyl;
[0313] R.sup.5 is selected from the group consisting of
C.sub.1-C.sub.4 alkyl, monocyclic aryl, and monocyclic aralkyl;
and
[0314] R.sup.6 is C.sub.1-C.sub.4 acyl;
[0315] R.sup.7 and R.sup.8 are independently selected from the
group consisting of hydrogen, C.sub.1-C.sub.4 acyl, C.sub.1-C.sub.4
alkoxycarbonyl, and a naturally-occurring L-amino acid connected
via its carbonyl group to form an ester; or
[0316] together R.sup.7 and R.sup.8 form a cyclic carbonate;
[0317] R.sup.9 is selected from the group consisting of amino,
azido, --N.dbd.CHN(R.sup.4).sub.2, --NHC(O)R.sup.4, and
--NHC(O)OR.sup.4; and
[0318] R.sup.10 is selected from the group consisting of OR.sup.6,
halogen, and H.
[0319] In another aspect, the invention comprises compounds of
Formula II: ##STR36##
[0320] wherein:
[0321] B is selected from the group consisting of ##STR37##
[0322] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl;
[0323] W and W' are independently selected from the group
consisting of --H, methyl, and V, or W and W' are each methyl, with
the proviso that when W is V, then W' is H;
[0324] Z is selected from the group consisting of --H, --OMe,
--OEt, phenyl, C.sub.1-C.sub.3 alkyl, --NR.sup.4.sub.2, --SR.sup.4,
--(CH.sub.2).sub.p--OR.sup.6, --(CH.sub.2).sub.p--SR.sup.6 and
--OCOR.sup.5; or
[0325] together V and Z are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing 1 heteroatom, that is
fused to an aryl group at the beta and gamma position to the O
attached to the phosphorus; or
[0326] together Z and W are connected via an additional 3-5 atoms
to form a cyclic group, optionally containing one heteroatom;
or
[0327] together W and W' are connected via an additional 2-5 atoms
to form a cyclic group;
[0328] R.sup.4 is C.sub.1-C.sub.4 alkyl;
[0329] R.sup.5 is selected from the group consisting of
C.sub.1-C.sub.4 alkyl, monocyclic aryl, and monocyclic aralkyl;
and
[0330] R.sup.6 is C.sub.1-C.sub.4 acyl;
[0331] R.sup.7 and R.sup.8 are independently selected from the
group consisting of hydrogen, C.sub.1-C.sub.4 acyl, C.sub.1-C.sub.4
alkoxycarbonyl, and a naturally-occurring L-amino acid connected
via its carbonyl group to form an ester; or
[0332] together R.sup.7 and R.sup.9 form a cyclic carbonate;
and
[0333] R.sup.10 is selected from the group consisting of OR.sup.4,
OR.sup.6, halogen, and H.
[0334] A further aspect of the invention comprises compounds of
Formula III: ##STR38##
[0335] wherein:
[0336] V and the 5'oxymethylene group of the ribose sugar moiety
are cis to one another;
[0337] B is selected from the group consisting of: ##STR39##
[0338] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl;
[0339] R.sup.4 is C.sub.1-C.sub.4 alkyl;
[0340] R.sup.6 is C.sub.1-C.sub.4 acyl;
[0341] R.sup.7 and R.sup.8 are independently selected from the
group consisting of hydrogen, C.sub.1-C.sub.4 acyl, C.sub.1-C.sub.4
alkoxycarbonyl, and a naturally-occurring L-amino acid connected
via its carbonyl group to form an ester; or
[0342] together R.sup.7 and R.sup.8 form a cyclic carbonate;
[0343] R.sup.9 is selected from the group consisting of amino,
azido, --N.dbd.CHN(R.sup.4).sub.2, --NHC(O)R.sup.4, and
--NHC(O)OR.sup.4; and
[0344] R.sup.10 is selected from the group consisting of OR.sup.6,
halogen, and H.
[0345] In one aspect, V is selected from the group consisting of
phenyl, substituted phenyl with 1-3 substituents independently
selected from the group consisting of --Cl, --Br, --F,
C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN, monocyclic heteroaryl,
and substituted monocyclic heteroaryl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN. In another aspect, V is
selected from the group consisting of phenyl, 3-chlorophenyl,
3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl,
3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl, and 4-pyridyl.
[0346] In another aspect, the invention comprises compounds of
Formula III: ##STR40##
[0347] wherein:
[0348] V and the 5'oxymethylene group of the ribose sugar moiety
are cis to one another;
[0349] B is selected from the group consisting of ##STR41##
[0350] V is selected from the group consisting of optionally
substituted monocyclic aryl and optionally substituted monocyclic
heteroaryl;
[0351] R.sup.4 is C.sub.1-C.sub.4 alkyl;
[0352] R.sup.6 is C.sub.1-C.sub.4 acyl;
[0353] R.sup.7 and R.sup.8 are independently selected from the
group consisting of hydrogen, C.sub.1-C.sub.4 acyl, C.sub.1-C.sub.4
alkoxycarbonyl, and a naturally-occurring L-amino acid connected
via its carbonyl group to form an ester; or
[0354] together R.sup.7 and R.sup.8 form a cyclic carbonate;
and
[0355] R.sup.10 is selected from the group consisting of OR.sup.4,
OR.sup.6, NH.sub.2, NHR.sup.4, halogen, and H.
[0356] In one aspect, V is selected from the group consisting of
phenyl, substituted phenyl with 1-3 substituents independently
selected from the group consisting of --Cl, --Br, --F,
C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN, monocyclic heteroaryl,
and substituted monocyclic heteroaryl with 1-2 substituents
independently selected from the group consisting of --Cl, --Br,
--F, C.sub.1-C.sub.3 alkyl, --CF.sub.3, --COCH.sub.3, --OMe,
--NMe.sub.2, --OEt, --CO.sub.2t-butyl, --CO.sub.2NH.sub.2, --SMe,
--SO.sub.2Me, --SO.sub.2NH.sub.2 and --CN. In another aspect, V is
selected from the group consisting of phenyl, 3-chlorophenyl,
3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl,
3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl, and 4-pyridyl.
[0357] In a further aspect, the compounds of this invention are
compounds of Formula VI: ##STR42##
[0358] wherein X is selected from the group consisting of NH.sub.2,
NHCH.sub.3, N(CH.sub.3).sub.2, OCH.sub.3, and SCH.sub.3;
[0359] Y and Y' are independently O or NH;
[0360] V, W, and W' are independently hydrogen, alkyl, alkenyl,
alkynyl, aryl, alkaryl, each of which is optionally substituted;
and
[0361] Z is hydrogen, CHWOH, CHWOCOW', SW, or CH.sub.2aryl.
[0362] In another aspect, the invention comprises compounds of
Formula VII: ##STR43##
Formula VII
[0363] wherein B is selected from the group consisting of:
##STR44##
[0364] X is selected from the group consisting of NH.sub.2,
NHCH.sub.3, N(CH.sub.3).sub.2, OCH.sub.3, SCH.sub.3, OH, and
SH;
[0365] Y and Y' are independently O or NH;
[0366] R.sup.14 is independently selected from the group consisting
of H and NH.sub.2;
[0367] the heterocyclic base may be further substituted at any
position on the heterocyclic base with a substituent of a molecular
weight of less than 150 and selected from the group consisting of
halogen, alkyl, alkenyl, alkynyl, aryl, alkaryl, cycloalkyl, acyl,
and alkoxy, and wherein said substituents may be coupled to the
6-position of the heterocyclic base via a carbon, sulfur, oxygen,
or selenium;
[0368] V, W, and W' are independently hydrogen, alkyl, alkenyl,
alkynyl, aryl, alkaryl, each of which is optionally substituted;
and
[0369] Z is hydrogen, CHWOH, CHWOCOW', SW, or CH.sub.2aryl.
[0370] In another aspect, B is selected from the group consisting
of: ##STR45##
[0371] In yet another aspect, B is selected from the groups
consisting of: ##STR46##
[0372] In another aspect, X is NH.sub.2.
[0373] In a further aspect, the invention comprises: ##STR47##
[0374] In another aspect, the invention comprises: ##STR48##
[0375] In a further aspect, the invention comprises: ##STR49##
[0376] In another aspect, the invention comprises: ##STR50##
[0377] In a further aspect, the invention comprises: ##STR51##
[0378] In another aspect of the invention the compounds of this
invention have R-stereochemistry at the V-attached carbon and have
S-stereochemistry at the phosphorus center. In another aspect of
the invention the compounds of this invention have
S-stereochemistry at the V-attached carbon and have
R-stereochemistry at the phosphorus center.
[0379] In one aspect the following compounds are included in the
invention but the compounds are not limited to these illustrative
compounds.
[0380] The following prodrugs are preferred compounds of the
invention. The compounds are shown without depiction of
stereochemistry since the compounds are biologically active as the
diastereomeric mixture or as a single stereoisomer. Compounds named
in Table 1 are designated by numbers assigned to the variables of
formula using the following convention: M1.V.L1.L2. M1 is a
variable that represents nucleosides of Formula I which are
attached via 5'-hydroxyl group that is phosphorylated with a group
P(O)(O--CH(V)CH.sub.2CH.sub.2--O) to make compounds of Formula VI.
V is an aryl or heteroaryl group that has 2 substituents, L1 and
L2, at the designated positions. V may have additional
substituents. ##STR52## Variable M1: ##STR53## Variable V: Group
V1
[0381] 1) 2-(L1)-3(L2)-phenyl
[0382] 2) 2-(L1)-4(L2)-phenyl
[0383] 3) 2-(L1)-5(L2)-phenyl
[0384] 4) 2-(L1)-6(L2)-phenyl
[0385] 5) 3-(L1)-4(L2)-phenyl
[0386] 6) 3-(L1)-5(L2)-phenyl
[0387] 7) 3-(L1)-6(L2)-phenyl
[0388] 8) 2-(L1)-6(L2)-3-chlorophenyl
[0389] 9) 4-(L1)-5(L2)-3-chlorophenyl
Variable V: Group V2
[0390] 1) 2-(L1)-3(L2)-4-pyridyl
[0391] 2) 2-(L1)-5(L2)-4-pyridyl
[0392] 3) 2-(L1)-6(L2)-4-pyridyl
[0393] 4) 3-(L1)-5(L2)-4-pyridyl
[0394] 5) 3-(L1)-6(L2)-4-pyridyl
[0395] 6) 2-(L1)-4(L2)-3-pyridyl
[0396] 7) 2-(L1)-5(L2)-3-pyridyl
[0397] 8) 2-(L1)-6(L2)-3-pyridyl
[0398] 9) 4-(L1)-5(L2)-3-pyridyl
Variable V: Group V3
[0399] 1) 4-(L1)-6(L2)-3-pyridyl
[0400] 2) 5-(L1)-6(L2)-3-pyridyl
[0401] 3) 3-(L1)-4(L2)-2-pyridyl
[0402] 4) 3-(L1)-5(L2)-2-pyridyl
[0403] 5) 3-(L1)-6(L2)-2-pyridyl
[0404] 6) 4-(L1)-5(L2)-2-pyridyl
[0405] 7) 4-(L1)-6(L2)-2-pyridyl
[0406] 8) 3-(L1)-4(L2)-2-thienyl
[0407] 9) 3-(L1)-4(L2)-2-furanyl
Variable L1
[0408] 1) hydrogen
[0409] 2) chloro
[0410] 3) bromo
[0411] 4) fluoro
[0412] 5) methyl
[0413] 6) trifluoromethyl
[0414] 7) methoxy
[0415] 8) dimethylamino
[0416] 9) cyano
Variable L2
[0417] 1) hydrogen
[0418] 2) chloro
[0419] 3) bromo
[0420] 4) fluoro
[0421] 5) methyl
[0422] 6) trifluoromethyl
[0423] 7) methoxy
[0424] 8) dimethylamino
[0425] 9) cyano
[0426] Preferred compounds are compounds listed in Table 1 using
variables M1 and V1 and L1 and L2 listed in that order. For
example, compound 1.3.6.7 represents structure 1 of variable M1,
i.e., 7-deaza-2'-methyl adenosine; structure 3 of group V1, i.e.,
2-(L1)-5-(L2) phenyl; structure 6 of variable L1, i.e.,
trifluoromethyl; and structure 7 of variable L2, i.e., methoxy. The
compound 1.3.6.7. therefore is 7-deaza-2'-methyladenosine with the
P(O)(O--CH(V)CH.sub.2CH.sub.2O) attached to the 5'-primary hydroxyl
group being
{[1-(2-trifluoromethyl-5-methoxyphenyl)-1,3-propyl]phosphoryl.
[0427] Preferred compounds are also compounds listed in Table 1
using variables M1 and V2 wherein the four digit number represents
M1.V2.L1.L2.
[0428] Preferred compounds are also compounds listed in Table 1
using variables M1 and V3 wherein the four digit number represents
M1.V3.L1.L2. TABLE-US-00001 TABLE 1 1.1.1.1 1.1.1.2 1.1.1.3 1.1.1.4
1.1.1.5 1.1.1.6 1.1.1.7 1.1.1.8 1.1.1.9 1.1.2.1 1.1.2.2 1.1.2.3
1.1.2.4 1.1.2.5 1.1.2.6 1.1.2.7 1.1.2.8 1.1.2.9 1.1.3.1 1.1.3.2
1.1.3.3 1.1.3.4 1.1.3.5 1.1.3.6 1.1.3.7 1.1.3.8 1.1.3.9 1.1.4.1
1.1.4.2 1.1.4.3 1.1.4.4 1.1.4.5 1.1.4.6 1.1.4.7 1.1.4.8 1.1.4.9
1.1.5.1 1.1.5.2 1.1.5.3 1.1.5.4 1.1.5.5 1.1.5.6 1.1.5.7 1.1.5.8
1.1.5.9 1.1.6.1 1.1.6.2 1.1.6.3 1.1.6.4 1.1.6.5 1.1.6.6 1.1.6.7
1.1.6.8 1.1.6.9 1.1.7.1 1.1.7.2 1.1.7.3 1.1.7.4 1.1.7.5 1.1.7.6
1.1.7.7 1.1.7.8 1.1.7.9 1.1.8.1 1.1.8.2 1.1.8.3 1.1.8.4 1.1.8.5
1.1.8.6 1.1.8.7 1.1.8.8 1.1.8.9 1.1.9.1 1.1.9.2 1.1.9.3 1.1.9.4
1.1.9.5 1.1.9.6 1.1.9.7 1.1.9.8 1.1.9.9 1.2.1.1 1.2.1.2 1.2.1.3
1.2.1.4 1.2.1.5 1.2.1.6 1.2.1.7 1.2.1.8 1.2.1.9 1.2.2.1 1.2.2.2
1.2.2.3 1.2.2.4 1.2.2.5 1.2.2.6 1.2.2.7 1.2.2.8 1.2.2.9 1.2.3.1
1.2.3.2 1.2.3.3 1.2.3.4 1.2.3.5 1.2.3.6 1.2.3.7 1.2.3.8 1.2.3.9
1.2.4.1 1.2.4.2 1.2.4.3 1.2.4.4 1.2.4.5 1.2.4.6 1.2.4.7 1.2.4.8
1.2.4.9 1.2.5.1 1.2.5.2 1.2.5.3 1.2.5.4 1.2.5.5 1.2.5.6 1.2.5.7
1.2.5.8 1.2.5.9 1.2.6.1 1.2.6.2 1.2.6.3 1.2.6.4 1.2.6.5 1.2.6.6
1.2.6.7 1.2.6.8 1.2.6.9 1.2.7.1 1.2.7.2 1.2.7.3 1.2.7.4 1.2.7.5
1.2.7.6 1.2.7.7 1.2.7.8 1.2.7.9 1.2.8.1 1.2.8.2 1.2.8.3 1.2.8.4
1.2.8.5 1.2.8.6 1.2.8.7 1.2.8.8 1.2.8.9 1.2.9.1 1.2.9.2 1.2.9.3
1.2.9.4 1.2.9.5 1.2.9.6 1.2.9.7 1.2.9.8 1.2.9.9 1.3.1.1 1.3.1.2
1.3.1.3 1.3.1.4 1.3.1.5 1.3.1.6 1.3.1.7 1.3.1.8 1.3.1.9 1.3.2.1
1.3.2.2 1.3.2.3 1.3.2.4 1.3.2.5 1.3.2.6 1.3.2.7 1.3.2.8 1.3.2.9
1.3.3.1 1.3.3.2 1.3.3.3 1.3.3.4 1.3.3.5 1.3.3.6 1.3.3.7 1.3.3.8
1.3.3.9 1.3.4.1 1.3.4.2 1.3.4.3 1.3.4.4 1.3.4.5 1.3.4.6 1.3.4.7
1.3.4.8 1.3.4.9 1.3.5.1 1.3.5.2 1.3.5.3 1.3.5.4 1.3.5.5 1.3.5.6
1.3.5.7 1.3.5.8 1.3.5.9 1.3.6.1 1.3.6.2 1.3.6.3 1.3.6.4 1.3.6.5
1.3.6.6 1.3.6.7 1.3.6.8 1.3.6.9 1.3.7.1 1.3.7.2 1.3.7.3 1.3.7.4
1.3.7.5 1.3.7.6 1.3.7.7 1.3.7.8 1.3.7.9 1.3.8.1 1.3.8.2 1.3.8.3
1.3.8.4 1.3.8.5 1.3.8.6 1.3.8.7 1.3.8.8 1.3.8.9 1.3.9.1 1.3.9.2
1.3.9.3 1.3.9.4 1.3.9.5 1.3.9.6 1.3.9.7 1.3.9.8 1.3.9.9 1.4.1.1
1.4.1.2 1.4.1.3 1.4.1.4 1.4.1.5 1.4.1.6 1.4.1.7 1.4.1.8 1.4.1.9
1.4.2.1 1.4.2.2 1.4.2.3 1.4.2.4 1.4.2.5 1.4.2.6 1.4.2.7 1.4.2.8
1.4.2.9 1.4.3.1 1.4.3.2 1.4.3.3 1.4.3.4 1.4.3.5 1.4.3.6 1.4.3.7
1.4.3.8 1.4.3.9 1.4.4.1 1.4.4.2 1.4.4.3 1.4.4.4 1.4.4.5 1.4.4.6
1.4.4.7 1.4.4.8 1.4.4.9 1.4.5.1 1.4.5.2 1.4.5.3 1.4.5.4 1.4.5.5
1.4.5.6 1.4.5.7 1.4.5.8 1.4.5.9 1.4.6.1 1.4.6.2 1.4.6.3 1.4.6.4
1.4.6.5 1.4.6.6 1.4.6.7 1.4.6.8 1.4.6.9 1.4.7.1 1.4.7.2 1.4.7.3
1.4.7.4 1.4.7.5 1.4.7.6 1.4.7.7 1.4.7.8 1.4.7.9 1.4.8.1 1.4.8.2
1.4.8.3 1.4.8.4 1.4.8.5 1.4.8.6 1.4.8.7 1.4.8.8 1.4.8.9 1.4.9.1
1.4.9.2 1.4.9.3 1.4.9.4 1.4.9.5 1.4.9.6 1.4.9.7 1.4.9.8 1.4.9.9
1.5.1.1 1.5.1.2 1.5.1.3 1.5.1.4 1.5.1.5 1.5.1.6 1.5.1.7 1.5.1.8
1.5.1.9 1.5.2.1 1.5.2.2 1.5.2.3 1.5.2.4 1.5.2.5 1.5.2.6 1.5.2.7
1.5.2.8 1.5.2.9 1.5.3.1 1.5.3.2 1.5.3.3 1.5.3.4 1.5.3.5 1.5.3.6
1.5.3.7 1.5.3.8 1.5.3.9 1.5.4.1 1.5.4.2 1.5.4.3 1.5.4.4 1.5.4.5
1.5.4.6 1.5.4.7 1.5.4.8 1.5.4.9 1.5.5.1 1.5.5.2 1.5.5.3 1.5.5.4
1.5.5.5 1.5.5.6 1.5.5.7 1.5.5.8 1.5.5.9 1.5.6.1 1.5.6.2 1.5.6.3
1.5.6.4 1.5.6.5 1.5.6.6 1.5.6.7 1.5.6.8 1.5.6.9 1.5.7.1 1.5.7.2
1.5.7.3 1.5.7.4 1.5.7.5 1.5.7.6 1.5.7.7 1.5.7.8 1.5.7.9 1.5.8.1
1.5.8.2 1.5.8.3 1.5.8.4 1.5.8.5 1.5.8.6 1.5.8.7 1.5.8.8 1.5.8.9
1.5.9.1 1.5.9.2 1.5.9.3 1.5.9.4 1.5.9.5 1.5.9.6 1.5.9.7 1.5.9.8
1.5.9.9 1.6.1.1 1.6.1.2 1.6.1.3 1.6.1.4 1.6.1.5 1.6.1.6 1.6.1.7
1.6.1.8 1.6.1.9 1.6.2.1 1.6.2.2 1.6.2.3 1.6.2.4 1.6.2.5 1.6.2.6
1.6.2.7 1.6.2.8 1.6.2.9 1.6.3.1 1.6.3.2 1.6.3.3 1.6.3.4 1.6.3.5
1.6.3.6 1.6.3.7 1.6.3.8 1.6.3.9 1.6.4.1 1.6.4.2 1.6.4.3 1.6.4.4
1.6.4.5 1.6.4.6 1.6.4.7 1.6.4.8 1.6.4.9 1.6.5.1 1.6.5.2 1.6.5.3
1.6.5.4 1.6.5.5 1.6.5.6 1.6.5.7 1.6.5.8 1.6.5.9 1.6.6.1 1.6.6.2
1.6.6.3 1.6.6.4 1.6.6.5 1.6.6.6 1.6.6.7 1.6.6.8 1.6.6.9 1.6.7.1
1.6.7.2 1.6.7.3 1.6.7.4 1.6.7.5 1.6.7.6 1.6.7.7 1.6.7.8 1.6.7.9
1.6.8.1 1.6.8.2 1.6.8.3 1.6.8.4 1.6.8.5 1.6.8.6 1.6.8.7 1.6.8.8
1.6.8.9 1.6.9.1 1.6.9.2 1.6.9.3 1.6.9.4 1.6.9.5 1.6.9.6 1.6.9.7
1.6.9.8 1.6.9.9 1.7.1.1 1.7.1.2 1.7.1.3 1.7.1.4 1.7.1.5 1.7.1.6
1.7.1.7 1.7.1.8 1.7.1.9 1.7.2.1 1.7.2.2 1.7.2.3 1.7.2.4 1.7.2.5
1.7.2.6 1.7.2.7 1.7.2.8 1.7.2.9 1.7.3.1 1.7.3.2 1.7.3.3 1.7.3.4
1.7.3.5 1.7.3.6 1.7.3.7 1.7.3.8 1.7.3.9 1.7.4.1 1.7.4.2 1.7.4.3
1.7.4.4 1.7.4.5 1.7.4.6 1.7.4.7 1.7.4.8 1.7.4.9 1.7.5.1 1.7.5.2
1.7.5.3 1.7.5.4 1.7.5.5 1.7.5.6 1.7.5.7 1.7.5.8 1.7.5.9 1.7.6.1
1.7.6.2 1.7.6.3 1.7.6.4 1.7.6.5 1.7.6.6 1.7.6.7 1.7.6.8 1.7.6.9
1.7.7.1 1.7.7.2 1.7.7.3 1.7.7.4 1.7.7.5 1.7.7.6 1.7.7.7 1.7.7.8
1.7.7.9 1.7.8.1 1.7.8.2 1.7.8.3 1.7.8.4 1.7.8.5 1.7.8.6 1.7.8.7
1.7.8.8 1.7.8.9 1.7.9.1 1.7.9.2 1.7.9.3 1.7.9.4 1.7.9.5 1.7.9.6
1.7.9.7 1.7.9.8 1.7.9.9 1.8.1.1 1.8.1.2 1.8.1.3 1.8.1.4 1.8.1.5
1.8.1.6 1.8.1.7 1.8.1.8 1.8.1.9 1.8.2.1 1.8.2.2 1.8.2.3 1.8.2.4
1.8.2.5 1.8.2.6 1.8.2.7 1.8.2.8 1.8.2.9 1.8.3.1 1.8.3.2 1.8.3.3
1.8.3.4 1.8.3.5 1.8.3.6 1.8.3.7 1.8.3.8 1.8.3.9 1.8.4.1 1.8.4.2
1.8.4.3 1.8.4.4 1.8.4.5 1.8.4.6 1.8.4.7 1.8.4.8 1.8.4.9 1.8.5.1
1.8.5.2 1.8.5.3 1.8.5.4 1.8.5.5 1.8.5.6 1.8.5.7 1.8.5.8 1.8.5.9
1.8.6.1 1.8.6.2 1.8.6.3 1.8.6.4 1.8.6.5 1.8.6.6 1.8.6.7 1.8.6.8
1.8.6.9 1.8.7.1 1.8.7.2 1.8.7.3 1.8.7.4 1.8.7.5 1.8.7.6 1.8.7.7
1.8.7.8 1.8.7.9 1.8.8.1 1.8.8.2 1.8.8.3 1.8.8.4 1.8.8.5 1.8.8.6
1.8.8.7 1.8.8.8 1.8.8.9 1.8.9.1 1.8.9.2 1.8.9.3 1.8.9.4 1.8.9.5
1.8.9.6 1.8.9.7 1.8.9.8 1.8.9.9 1.9.1.1 1.9.1.2 1.9.1.3 1.9.1.4
1.9.1.5 1.9.1.6 1.9.1.7 1.9.1.8 1.9.1.9 1.9.2.1 1.9.2.2 1.9.2.3
1.9.2.4 1.9.2.5 1.9.2.6 1.9.2.7 1.9.2.8 1.9.2.9 1.9.3.1 1.9.3.2
1.9.3.3 1.9.3.4 1.9.3.5 1.9.3.6 1.9.3.7 1.9.3.8 1.9.3.9 1.9.4.1
1.9.4.2 1.9.4.3 1.9.4.4 1.9.4.5 1.9.4.6 1.9.4.7 1.9.4.8 1.9.4.9
1.9.5.1 1.9.5.2 1.9.5.3 1.9.5.4 1.9.5.5 1.9.5.6 1.9.5.7 1.9.5.8
1.9.5.9 1.9.6.1 1.9.6.2 1.9.6.3 1.9.6.4 1.9.6.5 1.9.6.6 1.9.6.7
1.9.6.8 1.9.6.9 1.9.7.1 1.9.7.2 1.9.7.3 1.9.7.4 1.9.7.5 1.9.7.6
1.9.7.7 1.9.7.8 1.9.7.9 1.9.8.1 1.9.8.2 1.9.8.3 1.9.8.4 1.9.8.5
1.9.8.6 1.9.8.7 1.9.8.8 1.9.8.9 1.9.9.1 1.9.9.2 1.9.9.3 1.9.9.4
1.9.9.5 1.9.9.6 1.9.9.7 1.9.9.8 1.9.9.9 2.1.1.1 2.1.1.2 2.1.1.3
2.1.1.4 2.1.1.5 2.1.1.6 2.1.1.7 2.1.1.8 2.1.1.9 2.1.2.1 2.1.2.2
2.1.2.3 2.1.2.4 2.1.2.5 2.1.2.6 2.1.2.7 2.1.2.8 2.1.2.9 2.1.3.1
2.1.3.2 2.1.3.3 2.1.3.4 2.1.3.5 2.1.3.6 2.1.3.7 2.1.3.8 2.1.3.9
2.1.4.1 2.1.4.2 2.1.4.3 2.1.4.4 2.1.4.5 2.1.4.6 2.1.4.7 2.1.4.8
2.1.4.9 2.1.5.1 2.1.5.2 2.1.5.3 2.1.5.4 2.1.5.5 2.1.5.6 2.1.5.7
2.1.5.8 2.1.5.9 2.1.6.1 2.1.6.2 2.1.6.3 2.1.6.4 2.1.6.5 2.1.6.6
2.1.6.7 2.1.6.8 2.1.6.9 2.1.7.1 2.1.7.2 2.1.7.3 2.1.7.4 2.1.7.5
2.1.7.6 2.1.7.7 2.1.7.8 2.1.7.9 2.1.8.1 2.1.8.2 2.1.8.3 2.1.8.4
2.1.8.5 2.1.8.6 2.1.8.7 2.1.8.8 2.1.8.9 2.1.9.1 2.1.9.2 2.1.9.3
2.1.9.4 2.1.9.5 2.1.9.6 2.1.9.7 2.1.9.8 2.1.9.9 2.2.1.1 2.2.1.2
2.2.1.3 2.2.1.4 2.2.1.5 2.2.1.6 2.2.1.7 2.2.1.8 2.2.1.9 2.2.2.1
2.2.2.2 2.2.2.3 2.2.2.4 2.2.5.2 2.2.6.2 2.2.7.2 2.2.3.8 2.2.2.9
2.2.3.1 2.2.3.2 2.2.3.3 2.2.3.4 2.2.3.5 2.3.6.2 2.3.7.2 2.3.8.2
2.2.3.9 2.2.4.1 2.2.4.2 2.2.4.3 2.2.4.4 2.2.4.5 2.2.4.6 2.4.7.2
2.2.4.8 2.2.4.9 2.2.5.1 2.2.5.2 2.2.5.3 2.2.5.4 2.2.5.5 2.2.5.6
2.2.5.7 2.2.5.8 2.2.5.9 2.2.6.1 2.2.6.2 2.2.6.3 2.2.6.4 2.2.6.5
2.2.6.6 2.2.6.7 2.2.6.8 2.2.6.9 2.2.7.1 2.2.7.2 2.2.7.3 2.2.7.4
2.2.7.5 2.2.7.6 2.2.7.7 2.2.7.8 2.2.7.9 2.2.8.1 2.2.8.2 2.2.8.3
2.2.8.4 2.2.8.5 2.2.8.6 2.2.8.7 2.2.8.8 2.2.8.9 2.2.9.1 2.2.9.2
2.2.9.3 2.2.9.4 2.2.9.5 2.2.9.6 2.2.9.7 2.2.9.8 2.2.9.9 2.3.1.1
2.3.1.2 2.3.1.3 2.3.1.4 2.3.1.5 2.3.1.6 2.3.1.7 2.3.1.8 2.3.1.9
2.3.2.1 2.3.2.2 2.3.2.3 2.3.2.4 2.3.2.5 2.3.2.6 2.3.2.7 2.3.2.8
2.3.2.9 2.3.3.1 2.3.3.2 2.3.3.3 2.3.3.4 2.3.3.5 2.3.3.6 2.3.3.7
2.3.3.8 2.3.3.9 2.3.4.1 2.3.4.2 2.3.4.3 2.3.4.4 2.3.4.5 2.3.4.6
2.3.4.7 2.3.4.8 2.3.4.9 2.3.5.1 2.3.5.2 2.3.5.3 2.3.5.4 2.3.5.5
2.3.5.6 2.3.5.7 2.3.5.8 2.3.5.9 2.3.6.1 2.3.6.2 2.3.6.3 2.3.6.4
2.3.6.5 2.3.6.6 2.3.6.7 2.3.6.8 2.3.6.9 2.3.7.1 2.3.7.2 2.3.7.3
2.3.7.4 2.3.7.5 2.3.7.6 2.3.7.7 2.3.7.8 2.3.7.9 2.3.8.1 2.3.8.2
2.3.8.3 2.3.8.4 2.3.8.5 2.3.8.6 2.3.8.7 2.3.8.8 2.3.8.9 2.3.9.1
2.3.9.2 2.3.9.3 2.3.9.4 2.3.9.5 2.3.9.6 2.3.9.7 2.3.9.8 2.3.9.9
2.4.1.1 2.4.1.2 2.4.1.3 2.4.1.4 2.4.1.5 2.4.1.6 2.4.1.7 2.4.1.8
2.4.1.9 2.4.2.1 2.4.2.2 2.4.2.3 2.4.2.4 2.4.2.5 2.4.2.6 2.4.2.7
2.4.2.8 2.4.2.9 2.4.3.1 2.4.3.2 2.4.3.3 2.4.3.4 2.4.3.5 2.4.3.6
2.4.3.7 2.4.3.8 2.4.3.9 2.4.4.1 2.4.4.2 2.4.4.3 2.4.4.4 2.4.4.5
2.4.4.6 2.4.4.7 2.4.4.8 2.4.4.9 2.4.5.1 2.4.5.2 2.4.5.3 2.4.5.4
2.4.5.5 2.4.5.6 2.4.5.7 2.4.5.8 2.4.5.9 2.4.6.1 2.4.6.2 2.4.6.3
2.4.6.4 2.4.6.5 2.4.6.6 2.4.6.7 2.4.6.8 2.4.6.9 2.4.7.1 2.4.7.2
2.4.7.3 2.4.7.4 2.4.7.5 2.4.7.6 2.4.7.7 2.4.7.8 2.4.7.9 2.4.8.1
2.4.8.2 2.4.8.3 2.4.8.4 2.4.8.5 2.4.8.6 2.4.8.7 2.4.8.8 2.4.8.9
2.4.9.1 2.4.9.2 2.4.9.3 2.4.9.4 2.4.9.5 2.4.9.6 2.4.9.7 2.4.9.8
2.4.9.9 2.5.1.1 2.5.1.2 2.5.1.3 2.5.1.4 2.5.1.5 2.5.1.6 2.5.1.7
2.5.1.8 2.5.1.9 2.5.2.1 2.5.2.2 2.5.2.3 2.5.2.4 2.5.2.5 2.5.2.6
2.5.2.7 2.5.2.8 2.5.2.9 2.5.3.1 2.5.3.2 2.5.3.3 2.5.3.4 2.5.3.5
2.5.3.6 2.5.3.7 2.5.3.8 2.5.3.9 2.5.4.1 2.5.4.2 2.5.4.3 2.5.4.4
2.5.4.5 2.5.4.6 2.5.4.7 2.5.4.8 2.5.4.9 2.5.5.1 2.5.5.2 2.5.5.3
2.5.5.4 2.5.5.5 2.5.5.6 2.5.5.7 2.5.5.8 2.5.5.9 2.5.6.1 2.5.6.2
2.5.6.3 2.5.6.4 2.5.6.5 2.5.6.6 2.5.6.7 2.5.6.8 2.5.6.9 2.5.7.1
2.5.7.2 2.5.7.3 2.5.7.4 2.5.7.5 2.5.7.6 2.5.7.7 2.5.7.8 2.5.7.9
2.5.8.1 2.5.8.2 2.5.8.3 2.5.8.4 2.5.8.5 2.5.8.6 2.5.8.7 2.5.8.8
2.5.8.9 2.5.9.1 2.5.9.2 2.5.9.3 2.5.9.4 2.5.9.5 2.5.9.6 2.5.9.7
2.5.9.8 2.5.9.9 2.6.1.1 2.6.1.2 2.6.1.3 2.6.1.4 2.6.1.5 2.6.1.6
2.6.1.7 2.6.1.8 2.6.1.9 2.6.2.1 2.6.2.2 2.6.2.3 2.6.2.4 2.6.2.5
2.6.2.6 2.6.2.7 2.6.2.8 2.6.2.9 2.6.3.1 2.6.3.2 2.6.3.3 2.6.3.4
2.6.3.5 2.6.3.6 2.6.3.7 2.6.3.8 2.6.3.9 2.6.4.1 2.6.4.2 2.6.4.3
2.6.4.4 2.6.4.5 2.6.4.6 2.6.4.7 2.6.4.8 2.6.4.9 2.6.5.1 2.6.5.2
2.6.5.3 2.6.5.4 2.6.5.5 2.6.5.6 2.6.5.7 2.6.5.8 2.6.5.9 2.6.6.1
2.6.6.2 2.6.6.3 2.6.6.4 2.6.6.5 2.6.6.6 2.6.6.7 2.6.6.8 2.6.6.9
2.6.7.1 2.6.7.2 2.6.7.3 2.6.7.4 2.6.7.5 2.6.7.6 2.6.7.7 2.6.7.8
2.6.7.9 2.6.8.1 2.6.8.2 2.6.8.3 2.6.8.4 2.6.8.5 2.6.8.6 2.6.8.7
2.6.8.8 2.6.8.9 2.6.9.1 2.6.9.2 2.6.9.3 2.6.9.4 2.6.9.5 2.6.9.6
2.6.9.7 2.6.9.8 2.6.9.9 2.7.1.1 2.7.1.2 2.7.1.3 2.7.1.4 2.7.1.5
2.7.1.6 2.7.1.7 2.7.1.8 2.7.1.9 2.7.2.1 2.7.2.2 2.7.2.3 2.7.2.4
2.7.2.5
2.7.2.6 2.7.2.7 2.7.2.8 2.7.2.9 2.7.3.1 2.7.3.2 2.7.3.3 2.7.3.4
2.7.3.5 2.7.3.6 2.7.3.7 2.7.3.8 2.7.3.9 2.7.4.1 2.7.4.2 2.7.4.3
2.7.4.4 2.7.4.5 2.7.4.6 2.7.4.7 2.7.4.8 2.7.4.9 2.7.5.1 2.7.5.2
2.7.5.3 2.7.5.4 2.7.5.5 2.7.5.6 2.7.5.7 2.7.5.8 2.7.5.9 2.7.6.1
2.7.6.2 2.7.6.3 2.7.6.4 2.7.6.5 2.7.6.6 2.7.6.7 2.7.6.8 2.7.6.9
2.7.7.1 2.7.7.2 2.7.7.3 2.7.7.4 2.7.7.5 2.7.7.6 2.7.7.7 2.7.7.8
2.7.7.9 2.7.8.1 2.7.8.2 2.7.8.3 2.7.8.4 2.7.8.5 2.7.8.6 2.7.8.7
2.7.8.8 2.7.8.9 2.7.9.1 2.7.9.2 2.7.9.3 2.7.9.4 2.7.9.5 2.7.9.6
2.7.9.7 2.7.9.8 2.7.9.9 2.8.1.1 2.8.1.2 2.8.1.3 2.8.1.4 2.8.1.5
2.8.1.6 2.8.1.7 2.8.1.8 2.8.1.9 2.8.2.1 2.8.2.2 2.8.2.3 2.8.2.4
2.8.2.5 2.8.2.6 2.8.2.7 2.8.2.8 2.8.2.9 2.8.3.1 2.8.3.2 2.8.3.3
2.8.3.4 2.8.3.5 2.8.3.6 2.8.3.7 2.8.3.8 2.8.3.9 2.8.4.1 2.8.4.2
2.8.4.3 2.8.4.4 2.8.4.5 2.8.4.6 2.8.4.7 2.8.4.8 2.8.4.9 2.8.5.1
2.8.5.2 2.8.5.3 2.8.5.4 2.8.5.5 2.8.5.6 2.8.5.7 2.8.5.8 2.8.5.9
2.8.6.1 2.8.6.2 2.8.6.3 2.8.6.4 2.8.6.5 2.8.6.6 2.8.6.7 2.8.6.8
2.8.6.9 2.8.7.1 2.8.7.2 2.8.7.3 2.8.7.4 2.8.7.5 2.8.7.6 2.8.7.7
2.8.7.8 2.8.7.9 2.8.8.1 2.8.8.2 2.8.8.3 2.8.8.4 2.8.8.5 2.8.8.6
2.8.8.7 2.8.8.8 2.8.8.9 2.8.9.1 2.8.9.2 2.8.9.3 2.8.9.4 2.8.9.5
2.8.9.6 2.8.9.7 2.8.9.8 2.8.9.9 2.9.1.1 2.9.1.2 2.9.1.3 2.9.1.4
2.9.1.5 2.9.1.6 2.9.1.7 2.9.1.8 2.9.1.9 2.9.2.1 2.9.2.2 2.9.2.3
2.9.2.4 2.9.2.5 2.9.2.6 2.9.2.7 2.9.2.8 2.9.2.9 2.9.3.1 2.9.3.2
2.9.3.3 2.9.3.4 2.9.3.5 2.9.3.6 2.9.3.7 2.9.3.8 2.9.3.9 2.9.4.1
2.9.4.2 2.9.4.3 2.9.4.4 2.9.4.5 2.9.4.6 2.9.4.7 2.9.4.8 2.9.4.9
2.9.5.1 2.9.5.2 2.9.5.3 2.9.5.4 2.9.5.5 2.9.5.6 2.9.5.7 2.9.5.8
2.9.5.9 2.9.6.1 2.9.6.2 2.9.6.3 2.9.6.4 2.9.6.5 2.9.6.6 2.9.6.7
2.9.6.8 2.9.6.9 2.9.7.1 2.9.7.2 2.9.7.3 2.9.7.4 2.9.7.5 2.9.7.6
2.9.7.7 2.9.7.8 2.9.7.9 2.9.8.1 2.9.8.2 2.9.8.3 2.9.8.4 2.9.8.5
2.9.8.6 2.9.8.7 2.9.8.8 2.9.8.9 2.9.9.1 2.9.9.2 2.9.9.3 2.9.9.4
2.9.9.5 2.9.9.6 2.9.9.7 2.9.9.8 2.9.9.9 3.1.1.1 3.1.1.2 3.1.1.3
3.1.1.4 3.1.1.5 3.1.1.6 3.1.1.7 3.1.1.8 3.1.1.9 3.1.2.1 3.1.2.2
3.1.2.3 3.1.2.4 3.1.2.5 3.1.2.6 3.1.2.7 3.1.2.8 3.1.2.9 3.1.3.1
3.1.3.2 3.1.3.3 3.1.3.4 3.1.3.5 3.1.3.6 3.1.3.7 3.1.3.8 3.1.3.9
3.1.4.1 3.1.4.2 3.1.4.3 3.1.4.4 3.1.4.5 3.1.4.6 3.1.4.7 3.1.4.8
3.1.4.9 3.1.5.1 3.1.5.2 3.1.5.3 3.1.5.4 3.1.5.5 3.1.5.6 3.1.5.7
3.1.5.8 3.1.5.9 3.1.6.1 3.1.6.2 3.1.6.3 3.1.6.4 3.1.6.5 3.1.6.6
3.1.6.7 3.1.6.8 3.1.6.9 3.1.7.1 3.1.7.2 3.1.7.3 3.1.7.4 3.1.7.5
3.1.7.6 3.1.7.7 3.1.7.8 3.1.7.9 3.1.8.1 3.1.8.2 3.1.8.3 3.1.8.4
3.1.8.5 3.1.8.6 3.1.8.7 3.1.8.8 3.1.8.9 3.1.9.1 3.1.9.2 3.1.9.3
3.1.9.4 3.1.9.5 3.1.9.6 3.1.9.7 3.1.9.8 3.1.9.9 3.2.1.1 3.2.1.2
3.2.1.3 3.2.1.4 3.2.1.5 3.2.1.6 3.2.1.7 3.2.1.8 3.2.1.9 3.2.2.1
3.2.2.2 3.2.2.3 3.2.2.4 3.2.2.5 3.2.2.6 3.2.2.7 3.2.2.8 3.2.2.9
3.2.3.1 3.2.3.2 3.2.3.3 3.2.3.4 3.2.3.5 3.2.3.6 3.2.3.7 3.2.3.8
3.2.3.9 3.2.4.1 3.2.4.2 3.2.4.3 3.2.4.4 3.2.4.5 3.2.4.6 3.2.4.7
3.2.4.8 3.2.4.9 3.2.5.1 3.2.5.2 3.2.5.3 3.2.5.4 3.2.5.5 3.2.5.6
3.2.5.7 3.2.5.8 3.2.5.9 3.2.6.1 3.2.6.2 3.2.6.3 3.2.6.4 3.2.6.5
3.2.6.6 3.2.6.7 3.2.6.8 3.2.6.9 3.2.7.1 3.2.7.2 3.2.7.3 3.2.7.4
3.2.7.5 3.2.7.6 3.2.7.7 3.2.7.8 3.2.7.9 3.2.8.1 3.2.8.2 3.2.8.3
3.2.8.4 3.2.8.5 3.2.8.6 3.2.8.7 3.2.8.8 3.2.8.9 3.2.9.1 3.2.9.2
3.2.9.3 3.2.9.4 3.2.9.5 3.2.9.6 3.2.9.7 3.2.9.8 3.2.9.9 3.3.1.1
3.3.1.2 3.3.1.3 3.3.1.4 3.3.1.5 3.3.1.6 3.3.1.7 3.3.1.8 3.3.1.9
3.3.2.1 3.3.2.2 3.3.2.3 3.3.2.4 3.3.2.5 3.3.2.6 3.3.2.7 3.3.2.8
3.3.2.9 3.3.3.1 3.3.3.2 3.3.3.3 3.3.3.4 3.3.3.5 3.3.3.6 3.3.3.7
3.3.3.8 3.3.3.9 3.3.4.1 3.3.4.2 3.3.4.3 3.3.4.4 3.3.4.5 3.3.4.6
3.3.4.7 3.3.4.8 3.3.4.9 3.3.5.1 3.3.5.2 3.3.5.3 3.3.5.4 3.3.5.5
3.3.5.6 3.3.5.7 3.3.5.8 3.3.5.9 3.3.6.1 3.3.6.2 3.3.6.3 3.3.6.4
3.3.6.5 3.3.6.6 3.3.6.7 3.3.6.8 3.3.6.9 3.3.7.1 3.3.7.2 3.3.7.3
3.3.7.4 3.3.7.5 3.3.7.6 3.3.7.7 3.3.7.8 3.3.7.9 3.3.8.1 3.3.8.2
3.3.8.3 3.3.8.4 3.3.8.5 3.3.8.6 3.3.8.7 3.3.8.8 3.3.8.9 3.9.1.3
3.9.2.3 3.9.3.3 3.3.9.4 3.3.9.5 3.3.9.6 3.3.9.7 3.3.9.8 3.3.9.9
3.4.1.1 3.4.1.2 3.4.1.3 3.4.1.4 3.4.1.5 3.4.1.6 3.4.1.7 3.4.1.8
3.4.1.9 3.4.2.1 3.4.2.2 3.4.2.3 3.4.2.4 3.4.2.5 3.4.2.6 3.4.2.7
3.4.2.8 3.4.2.9 3.4.3.1 3.4.3.2 3.4.3.3 3.4.3.4 3.4.3.5 3.4.3.6
3.4.3.7 3.4.3.8 3.4.3.9 3.4.4.1 3.4.4.2 3.4.4.3 3.4.4.4 3.4.4.5
3.4.4.6 3.4.4.7 3.4.4.8 3.4.4.9 3.4.5.1 3.4.5.2 3.4.5.3 3.4.5.4
3.4.5.5 3.4.5.6 3.4.5.7 3.4.5.8 3.4.5.9 3.4.6.1 3.4.6.2 3.4.6.3
3.4.6.4 3.4.6.5 3.4.6.6 3.4.6.7 3.4.6.8 3.4.6.9 3.4.7.1 3.4.7.2
3.4.7.3 3.4.7.4 3.4.7.5 3.4.7.6 3.4.7.7 3.4.7.8 3.4.7.9 3.4.8.1
3.4.8.2 3.4.8.3 3.4.8.4 3.4.8.5 3.4.8.6 3.4.8.7 3.4.8.8 3.4.8.9
3.4.9.1 3.4.9.2 3.4.9.3 3.4.9.4 3.4.9.5 3.4.9.6 3.4.9.7 3.4.9.8
3.4.9.9 3.5.1.1 3.5.1.2 3.5.1.3 3.5.1.4 3.5.1.5 3.5.1.6 3.5.1.7
3.5.1.8 3.5.1.9 3.5.2.1 3.5.2.2 3.5.2.3 3.5.2.4 3.5.2.5 3.5.2.6
3.5.2.7 3.5.2.8 3.5.2.9 3.5.3.1 3.5.3.2 3.5.3.3 3.5.3.4 3.5.3.5
3.5.3.6 3.5.3.7 3.5.3.8 3.5.3.9 3.5.4.1 3.5.4.2 3.5.4.3 3.5.4.4
3.5.4.5 3.5.4.6 3.5.4.7 3.5.4.8 3.5.4.9 3.5.5.1 3.5.5.2 3.5.5.3
3.5.5.4 3.5.5.5 3.5.5.6 3.5.5.7 3.5.5.8 3.5.5.9 3.5.6.1 3.5.6.2
3.5.6.3 3.5.6.4 3.5.6.5 3.5.6.6 3.5.6.7 3.5.6.8 3.5.6.9 3.5.7.1
3.5.7.2 3.5.7.3 3.5.7.4 3.5.7.5 3.5.7.6 3.5.7.7 3.5.7.8 3.5.7.9
3.5.8.1 3.5.8.2 3.5.8.3 3.5.8.4 3.5.8.5 3.5.8.6 3.5.8.7 3.5.8.8
3.5.8.9 3.5.9.1 3.5.9.2 3.5.9.3 3.5.9.4 3.5.9.5 3.5.9.6 3.5.9.7
3.5.9.8 3.5.9.9 3.6.1.1 3.6.1.2 3.6.1.3 3.6.1.4 3.6.1.5 3.6.1.6
3.6.1.7 3.6.1.8 3.6.1.9 3.6.2.1 3.6.2.2 3.6.2.3 3.6.2.4 3.6.2.5
3.6.2.6 3.6.2.7 3.6.2.8 3.6.2.9 3.6.3.1 3.6.3.2 3.6.3.3 3.6.3.4
3.6.3.5 3.6.3.6 3.6.3.7 3.6.3.8 3.6.3.9 3.6.4.1 3.6.4.2 3.6.4.3
3.6.4.4 3.6.4.5 3.6.4.6 3.6.4.7 3.6.4.8 3.6.4.9 3.6.5.1 3.6.5.2
3.6.5.3 3.6.5.4 3.6.5.5 3.6.5.6 3.6.5.7 3.6.5.8 3.6.5.9 3.6.6.1
3.6.6.2 3.6.6.3 3.6.6.4 3.6.6.5 3.6.6.6 3.6.6.7 3.6.6.8 3.6.6.9
3.6.7.1 3.6.7.2 3.6.7.3 3.6.7.4 3.6.7.5 3.6.7.6 3.6.7.7 3.6.7.8
3.6.7.9 3.6.8.1 3.6.8.2 3.6.8.3 3.6.8.4 3.6.8.5 3.6.8.6 3.6.8.7
3.6.8.8 3.6.8.9 3.6.9.1 3.6.9.2 3.6.9.3 3.6.9.4 3.6.9.5 3.6.9.6
3.6.9.7 3.6.9.8 3.6.9.9 3.7.1.1 3.7.1.2 3.7.1.3 3.7.1.4 3.7.1.5
3.7.1.6 3.7.1.7 3.7.1.8 3.7.1.9 3.7.2.1 3.7.2.2 3.7.2.3 3.7.2.4
3.7.2.5 3.7.2.6 3.7.2.7 3.7.2.8 3.7.2.9 3.7.3.1 3.7.3.2 3.7.3.3
3.7.3.4 3.7.3.5 3.7.3.6 3.7.3.7 3.7.3.8 3.7.3.9 3.7.4.1 3.7.4.2
3.7.4.3 3.7.4.4 3.7.4.5 3.7.4.6 3.7.4.7 3.7.4.8 3.7.4.9 3.7.5.1
3.7.5.2 3.7.5.3 3.7.5.4 3.7.5.5 3.7.5.6 3.7.5.7 3.7.5.8 3.7.5.9
3.7.6.1 3.7.6.2 3.7.6.3 3.7.6.4 3.7.6.5 3.7.6.6 3.7.6.7 3.7.6.8
3.7.6.9 3.7.7.1 3.7.7.2 3.7.7.3 3.7.7.4 3.7.7.5 3.7.7.6 3.7.7.7
3.7.7.8 3.7.7.9 3.7.8.1 3.7.8.2 3.7.8.3 3.7.8.4 3.7.8.5 3.7.8.6
3.7.8.7 3.7.8.8 3.7.8.9 3.7.9.1 3.7.9.2 3.7.9.3 3.7.9.4 3.7.9.5
3.7.9.6 3.7.9.7 3.7.9.8 3.7.9.9 3.8.1.1 3.8.1.2 3.8.1.3 3.8.1.4
3.8.1.5 3.8.1.6 3.8.1.7 3.8.1.8 3.8.1.9 3.8.2.1 3.8.2.2 3.8.2.3
3.8.2.4 3.8.2.5 3.8.2.6 3.8.2.7 3.8.2.8 3.8.2.9 3.8.3.1 3.8.3.2
3.8.3.3 3.8.3.4 3.8.3.5 3.8.3.6 3.8.3.7 3.8.3.8 3.8.3.9 3.8.4.1
3.8.4.2 3.8.4.3 3.8.4.4 3.8.4.5 3.8.4.6 3.8.4.7 3.8.4.8 3.8.4.9
3.8.5.1 3.8.5.2 3.8.5.3 3.8.5.4 3.8.5.5 3.8.5.6 3.8.5.7 3.8.5.8
3.8.5.9 3.8.6.1 3.8.6.2 3.8.6.3 3.8.6.4 3.8.6.5 3.8.6.6 3.8.6.7
3.8.6.8 3.8.6.9 3.8.7.1 3.8.7.2 3.8.7.3 3.8.7.4 3.8.7.5 3.8.7.6
3.8.7.7 3.8.7.8 3.8.7.9 3.8.8.1 3.8.8.2 3.8.8.3 3.8.8.4 3.8.8.5
3.8.8.6 3.8.8.7 3.8.8.8 3.8.8.9 3.8.9.1 3.8.9.2 3.8.9.3 3.8.9.4
3.8.9.5 3.8.9.6 3.8.9.7 3.8.9.8 3.8.9.9 3.9.1.1 3.9.1.2 3.9.1.3
3.9.1.4 3.9.1.5 3.9.1.6 3.9.1.7 3.9.1.8 3.9.1.9 3.9.2.1 3.9.2.2
3.9.2.3 3.9.2.4 3.9.2.5 3.9.2.6 3.9.2.7 3.9.2.8 3.9.2.9 3.9.3.1
3.9.3.2 3.9.3.3 3.9.3.4 3.9.3.5 3.9.3.6 3.9.3.7 3.9.3.8 3.9.3.9
3.9.4.1 3.9.4.2 3.9.4.3 3.9.4.4 3.9.4.5 3.9.4.6 3.9.4.7 3.9.4.8
3.9.4.9 3.9.5.1 3.9.5.2 3.9.5.3 3.9.5.4 3.9.5.5 3.9.5.6 3.9.5.7
3.9.5.8 3.9.5.9 3.9.6.1 3.9.6.2 3.9.6.3 3.9.6.4 3.9.6.5 3.9.6.6
3.9.6.7 3.9.6.8 3.9.6.9 3.9.7.1 3.9.7.2 3.9.7.3 3.9.7.4 3.9.7.5
3.9.7.6 3.9.7.7 3.9.7.8 3.9.7.9 3.9.8.1 3.9.8.2 3.9.8.3 3.9.8.4
3.9.8.5 3.9.8.6 3.9.8.7 3.9.8.8 3.9.8.9 3.9.9.1 3.9.9.2 3.9.9.3
3.9.9.4 3.9.9.5 3.9.9.6 3.9.9.7 3.9.9.8 3.9.9.9 4.1.1.1 4.1.1.2
4.1.1.3 4.1.1.4 4.1.1.5 4.1.1.6 4.1.1.7 4.1.1.8 4.1.1.9 4.1.2.1
4.1.2.2 4.1.2.3 4.1.2.4 4.1.2.5 4.1.2.6 4.1.2.7 4.1.2.8 4.1.2.9
4.1.3.1 4.1.3.2 4.1.3.3 4.1.3.4 4.1.3.5 4.1.3.6 4.1.3.7 4.1.3.8
4.1.3.9 4.1.4.1 4.1.4.2 4.1.4.3 4.1.4.4 4.1.4.5 4.1.4.6 4.1.4.7
4.1.4.8 4.1.4.9 4.1.5.1 4.1.5.2 4.1.5.3 4.1.5.4 4.1.5.5 4.1.5.6
4.1.5.7 4.1.5.8 4.1.5.9 4.1.6.1 4.1.6.2 4.1.6.3 4.1.6.4 4.1.6.5
4.1.6.6 4.1.6.7 4.1.6.8 4.1.6.9 4.1.7.1 4.1.7.2 4.1.7.3 4.1.7.4
4.1.7.5 4.1.7.6 4.1.7.7 4.1.7.8 4.1.7.9 4.1.8.1 4.1.8.2 4.1.8.3
4.1.8.4 4.1.8.5 4.1.8.6 4.1.8.7 4.1.8.8 4.1.8.9 4.1.9.1 4.1.9.2
4.1.9.3 4.1.9.4 4.1.9.5 4.1.9.6 4.1.9.7 4.1.9.8 4.1.9.9 4.2.1.1
4.2.1.2 4.2.1.3 4.2.1.4 4.2.1.5 4.2.1.6 4.2.1.7 4.2.1.8 4.2.1.9
4.2.2.1 4.2.2.2 4.2.2.3 4.2.2.4 4.2.2.5 4.2.2.6 4.2.2.7 4.2.2.8
4.2.2.9 4.2.3.1 4.2.3.2 4.2.3.3 4.2.3.4 4.2.3.5 4.2.3.6 4.2.3.7
4.2.3.8 4.2.3.9 4.2.4.1 4.2.4.2 4.2.4.3 4.2.4.4 4.2.4.5 4.2.4.6
4.2.4.7 4.2.4.8 4.2.4.9 4.2.5.1 4.2.5.2 4.2.5.3 4.2.5.4 4.2.5.5
4.2.5.6 4.2.5.7 4.2.5.8 4.2.5.9 4.2.6.1 4.2.6.2 4.2.6.3 4.2.6.4
4.2.6.5 4.2.6.6 4.2.6.7 4.2.6.8 4.2.6.9 4.2.7.1 4.2.7.2 4.2.7.3
4.2.7.4 4.2.7.5 4.2.7.6 4.2.7.7 4.2.7.8 4.2.7.9 4.2.8.1 4.2.8.2
4.2.8.3 4.2.8.4 4.2.8.5 4.2.8.6 4.2.8.7 4.2.8.8 4.2.8.9 4.2.9.1
4.2.9.2 4.2.9.3 4.2.9.4 4.2.9.5 4.2.9.6 4.2.9.7 4.2.9.8 4.2.9.9
4.3.1.1 4.3.1.2 4.3.1.3 4.3.1.4 4.3.1.5 4.3.1.6 4.3.1.7 4.3.1.8
4.3.1.9 4.3.2.1 4.3.2.2 4.3.2.3 4.3.2.4 4.3.2.5 4.3.2.6 4.3.2.7
4.3.2.8 4.3.2.9 4.3.3.1 4.3.3.2 4.3.3.3 4.3.3.4 4.3.3.5 4.3.3.6
4.3.3.7 4.3.3.8 4.3.3.9 4.3.4.1 4.3.4.2 4.3.4.3 4.3.4.4 4.3.4.5
4.3.4.6 4.3.4.7 4.3.4.8 4.3.4.9 4.3.5.1 4.3.5.2 4.3.5.3 4.3.5.4
4.3.5.5 4.3.5.6 4.3.5.7 4.3.5.8 4.3.5.9 4.3.6.1 4.3.6.2 4.3.6.3
4.3.6.4 4.3.6.5 4.3.6.6 4.3.6.7 4.3.6.8 4.3.6.9 4.3.7.1 4.3.7.2
4.3.7.3 4.3.7.4 4.3.7.5 4.3.7.6 4.3.7.7 4.3.7.8 4.3.7.9 4.3.8.1
4.3.8.2 4.3.8.3 4.3.8.4 4.3.8.5 4.3.8.6 4.3.8.7 4.3.8.8 4.3.8.9
4.3.9.1 4.3.9.2 4.3.9.3 4.3.9.4 4.3.9.5 4.3.9.6 4.3.9.7 4.3.9.8
4.3.9.9 4.4.1.1 4.4.1.2 4.4.1.3 4.4.1.4 4.4.1.5 4.4.1.6 4.4.1.7
4.4.1.8 4.4.1.9 4.4.2.1 4.4.2.2 4.4.2.3 4.4.2.4 4.4.2.5 4.4.2.6
4.4.2.7 4.4.2.8 4.4.2.9 4.4.3.1 4.4.3.2 4.4.3.3 4.4.3.4 4.4.3.5
4.4.3.6 4.4.3.7 4.4.3.8 4.4.3.9 4.4.4.1 4.4.4.2 4.4.4.3 4.4.4.4
4.4.4.5 4.4.4.6 4.4.4.7 4.4.4.8 4.4.4.9 4.4.5.1 4.4.5.2 4.4.5.3
4.4.5.4 4.4.5.5 4.4.5.6 4.4.5.7 4.4.5.8 4.4.5.9 4.4.6.1 4.4.6.2
4.4.6.3 4.4.6.4 4.4.6.5
4.4.6.6 4.4.6.7 4.4.6.8 4.4.6.9 4.4.7.1 4.4.7.2 4.4.7.3 4.4.7.4
4.4.7.5 4.4.7.6 4.4.7.7 4.4.7.8 4.4.7.9 4.4.8.1 4.4.8.2 4.4.8.3
4.4.8.4 4.4.8.5 4.4.8.6 4.4.8.7 4.4.8.8 4.4.8.9 4.4.9.1 4.4.9.2
4.4.9.3 4.4.9.4 4.4.9.5 4.4.9.6 4.4.9.7 4.4.9.8 4.4.9.9 4.5.1.1
4.5.1.2 4.5.1.3 4.5.1.4 4.5.1.5 4.5.1.6 4.5.1.7 4.5.1.8 4.5.1.9
4.5.2.1 4.5.2.2 4.5.2.3 4.5.2.4 4.5.2.5 4.5.2.6 4.5.2.7 4.5.2.8
4.5.2.9 4.5.3.1 4.5.3.2 4.5.3.3 4.5.3.4 4.5.3.5 4.5.3.6 4.5.3.7
4.5.3.8 4.5.3.9 4.5.4.1 4.5.4.2 4.5.4.3 4.5.4.4 4.5.4.5 4.5.4.6
4.5.4.7 4.5.4.8 4.5.4.9 4.5.5.1 4.5.5.2 4.5.5.3 4.5.5.4 4.5.5.5
4.5.5.6 4.5.5.7 4.5.5.8 4.5.5.9 4.5.6.1 4.5.6.2 4.5.6.3 4.5.6.4
4.5.6.5 4.5.6.6 4.5.6.7 4.5.6.8 4.5.6.9 4.5.7.1 4.5.7.2 4.5.7.3
4.5.7.4 4.5.7.5 4.5.7.6 4.5.7.7 4.5.7.8 4.5.7.9 4.5.8.1 4.5.8.2
4.5.8.3 4.5.8.4 4.5.8.5 4.5.8.6 4.5.8.7 4.5.8.8 4.5.8.9 4.5.9.1
4.5.9.2 4.5.9.3 4.5.9.4 4.5.9.5 4.5.9.6 4.5.9.7 4.5.9.8 4.5.9.9
4.6.1.1 4.6.1.2 4.6.1.3 4.6.1.4 4.6.1.5 4.6.1.6 4.6.1.7 4.6.1.8
4.6.1.9 4.6.2.1 4.6.2.2 4.6.2.3 4.6.2.4 4.6.2.5 4.6.2.6 4.6.2.7
4.6.2.8 4.6.2.9 4.6.3.1 4.6.3.2 4.6.3.3 4.6.3.4 4.6.3.5 4.6.3.6
4.6.3.7 4.6.3.8 4.6.3.9 4.6.4.1 4.6.4.2 4.6.4.3 4.6.4.4 4.6.4.5
4.6.4.6 4.6.4.7 4.6.4.8 4.6.4.9 4.6.5.1 4.6.5.2 4.6.5.3 4.6.5.4
4.6.5.5 4.6.5.6 4.6.5.7 4.6.5.8 4.6.5.9 4.6.6.1 4.6.6.2 4.6.6.3
4.6.6.4 4.6.6.5 4.6.6.6 4.6.6.7 4.6.6.8 4.6.6.9 4.6.7.1 4.6.7.2
4.6.7.3 4.6.7.4 4.6.7.5 4.6.7.6 4.6.7.7 4.6.7.8 4.6.7.9 4.6.8.1
4.6.8.2 4.6.8.3 4.6.8.4 4.6.8.5 4.6.8.6 4.6.8.7 4.6.8.8 4.6.8.9
4.6.9.1 4.6.9.2 4.6.9.3 4.6.9.4 4.6.9.5 4.6.9.6 4.6.9.7 4.6.9.8
4.6.9.9 4.7.1.1 4.7.1.2 4.7.1.3 4.7.1.4 4.7.1.5 4.7.1.6 4.7.1.7
4.7.1.8 4.7.1.9 4.7.2.1 4.7.2.2 4.7.2.3 4.7.2.4 4.7.2.5 4.7.2.6
4.7.2.7 4.7.2.8 4.7.2.9 4.7.3.1 4.7.3.2 4.7.3.3 4.7.3.4 4.7.3.5
4.7.3.6 4.7.3.7 4.7.3.8 4.7.3.9 4.7.4.1 4.7.4.2 4.7.4.3 4.7.4.4
4.7.4.5 4.7.4.6 4.7.4.7 4.7.4.8 4.7.4.9 4.7.5.1 4.7.5.2 4.7.5.3
4.7.5.4 4.7.5.5 4.7.5.6 4.7.5.7 4.7.5.8 4.7.5.9 4.7.6.1 4.7.6.2
4.7.6.3 4.7.6.4 4.7.6.5 4.7.6.6 4.7.6.7 4.7.6.8 4.7.6.9 4.7.7.1
4.7.7.2 4.7.7.3 4.7.7.4 4.7.7.5 4.7.7.6 4.7.7.7 4.7.7.8 4.7.7.9
4.7.8.1 4.7.8.2 4.7.8.3 4.7.8.4 4.7.8.5 4.7.8.6 4.7.8.7 4.7.8.8
4.7.8.9 4.7.9.1 4.7.9.2 4.7.9.3 4.7.9.4 4.7.9.5 4.7.9.6 4.7.9.7
4.7.9.8 4.7.9.9 4.8.1.1 4.8.1.2 4.8.1.3 4.8.1.4 4.8.1.5 4.8.1.6
4.8.1.7 4.8.1.8 4.8.1.9 4.8.2.1 4.8.2.2 4.8.2.3 4.8.2.4 4.8.2.5
4.8.2.6 4.8.2.7 4.8.2.8 4.8.2.9 4.8.3.1 4.8.3.2 4.8.3.3 4.8.3.4
4.8.3.5 4.8.3.6 4.8.3.7 4.8.3.8 4.8.3.9 4.8.4.1 4.8.4.2 4.8.4.3
4.8.4.4 4.8.4.5 4.8.4.6 4.8.4.7 4.8.4.8 4.8.4.9 4.8.5.1 4.8.5.2
4.8.5.3 4.8.5.4 4.8.5.5 4.8.5.6 4.8.5.7 4.8.5.8 4.8.5.9 4.8.6.1
4.8.6.2 4.8.6.3 4.8.6.4 4.8.6.5 4.8.6.6 4.8.6.7 4.8.6.8 4.8.6.9
4.8.7.1 4.8.7.2 4.8.7.3 4.8.7.4 4.8.7.5 4.8.7.6 4.8.7.7 4.8.7.8
4.8.7.9 4.8.8.1 4.8.8.2 4.8.8.3 4.8.8.4 4.8.8.5 4.8.8.6 4.8.8.7
4.8.8.8 4.8.8.9 4.8.9.1 4.8.9.2 4.8.9.3 4.8.9.4 4.8.9.5 4.8.9.6
4.8.9.7 4.8.9.8 4.8.9.9 4.9.1.1 4.9.1.2 4.9.1.3 4.9.1.4 4.9.1.5
4.9.1.6 4.9.1.7 4.9.1.8 4.9.1.9 4.9.2.1 4.9.2.2 4.9.2.3 4.9.2.4
4.9.2.5 4.9.2.6 4.9.2.7 4.9.2.8 4.9.2.9 4.9.3.1 4.9.3.2 4.9.3.3
4.9.3.4 4.9.3.5 4.9.3.6 4.9.3.7 4.9.3.8 4.9.3.9 4.9.4.1 4.9.4.2
4.9.4.3 4.9.4.4 4.9.4.5 4.9.4.6 4.9.4.7 4.9.4.8 4.9.4.9 4.9.5.1
4.9.5.2 4.9.5.3 4.9.5.4 4.9.5.5 4.9.5.6 4.9.5.7 4.9.5.8 4.9.5.9
4.9.6.1 4.9.6.2 4.9.6.3 4.9.6.4 4.9.6.5 4.9.6.6 4.9.6.7 4.9.6.8
4.9.6.9 4.9.7.1 4.9.7.2 4.9.7.3 4.9.7.4 4.9.7.5 4.9.7.6 4.9.7.7
4.9.7.8 4.9.7.9 4.9.8.1 4.9.8.2 4.9.8.3 4.9.8.4 4.9.8.5 4.9.8.6
4.9.8.7 4.9.8.8 4.9.8.9 4.9.9.1 4.9.9.2 4.9.9.3 4.9.9.4 4.9.9.5
4.9.9.6 4.9.9.7 4.9.9.8 4.9.9.9
[0429] Another group of preferred compounds are named in Table 2
and designated by numbers assigned to the variables of Formula I
using the following convention: M1.V/Z/W. The compounds are shown
without depiction of stereochemistry since the compounds are
biologically active as the diastereomeric mixture or as a single
stereoisomer. M1 is a variable that represents nucleosides of
Formula I which are attached via 5'-hydroxyl group that is
phosphorylated with a group P(O)(O--CH(V)CH(Z)C(WW')--O) to make
compounds of Formula I. ##STR54##
[0430] The structures for variable M1 are the same as described
above.
Variable V/Z/W: Group 1 of V/Z/W
[0431] 1) V=3-chlorophenyl; Z=methyl; W=hydrogen [0432] 2)
V=3,5-dichlorophenyl; Z=methyl; W=hydrogen [0433] 3) V=4-pyridyl;
Z=methyl; W=hydrogen [0434] 4) V=3-chlorophenyl; Z=methoxy;
W=hydrogen [0435] 5) V=3,5-dichlorophenyl; Z=methoxy; W=hydrogen
[0436] 6) V=4-pyridyl; Z=methoxy; W=hydrogen [0437] 7)
V=3-chlorophenyl; Z=hydrogen; W=3-chlorophenyl [0438] 8)
V=3,5-dichlorophenyl; Z=hydrogen; W=3,5-dichlorophenyl [0439] 9)
V=4-pyridyl; Z=hydrogen; W=4-pyridyl Variable V/Z/W: Group 2 of
V/Z/W [0440] 1) V=3-chlorophenyl; Z=NHAc; W=hydrogen [0441] 2)
V=3,5-dichlorophenyl; Z=NHAc; W=hydrogen [0442] 3) V=4-pyridyl;
Z=NHAc; W=hydrogen [0443] 4) V=3-chlorophenyl; Z=hydrogen; W=methyl
[0444] 5) V=3,5-dichlorophenyl; Z=hydrogen; W=methyl [0445] 6)
V=4-pyridyl; Z=hydrogen; W=methyl [0446] 7) V=3-chlorophenyl;
Z=acetoxy; W=hydrogen [0447] 8) V=3,5-dichlorophenyl; Z=acetoxy;
W=hydrogen [0448] 9) V=4-pyridyl; Z=acetoxy; W=hydrogen Variable
V/Z/W: Group 3 of V/Z/W [0449] 1) V=phenyl; Z=phenyl; W=hydrogen
[0450] 2) V=phenyl; Z=--CH.sub.2--CH.sub.2-- fused to phenyl at V
to form a 6-membered ring; W=hydrogen [0451] 3) V=phenyl; Z=H;
W=--CH.sub.2--CH.sub.2-- fused to phenyl at V to form a 6-membered
ring [0452] 4) V=phenyl; Z=H; W=W'=methyl [0453] 5) V=phenyl; Z=H;
W and W'=--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2-- to form a
6-membered ring [0454] 6) V=phenyl; Z and
W=--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2-- to form a 6-membered
ring [0455] 7) V=3-chlorophenyl;
Z=CH.sub.2CH.sub.2CH.sub.2OC(O)OCH.sub.3; W=hydrogen [0456] 8)
V=3-chlorophenyl; Z=CH.sub.2CH.sub.2CH.sub.2SC(O)CH.sub.3;
W=hydrogen [0457] 9) V=4-pyridyl;
Z=CH.sub.2CH.sub.2CH.sub.2OC(O)OCH.sub.3; W=hydrogen [0458] 10)
V=4-pyridyl; Z=CH.sub.2CH.sub.2CH.sub.2SC(O)CH.sub.3; W=hydrogen
[0459] W' is hydrogen when not specified.
[0460] Preferred compounds are compounds listed in Table 2 using
groups M1 and Group 1 of V/Z/W. For example, compound 1.3
represents structure 1 of group M1, i.e., 7-deaza-2'-methyl
adenosine; and structure 3 of Group 1 of V/Z/W, i.e., V=4-pyridyl,
Z=methyl and W=hydrogen. The compound 1.3 therefore is
7-deaza-2'-methyladenosine with the
P(O)(O--CH(4-pyridyl)CH(CH.sub.3)CH.sub.2O) attached to the primary
hydroxyl.
[0461] Preferred compounds are also compounds listed in Table 2
using groups M1 and Group 2 of V/Z/W.
[0462] Preferred compounds are also compounds listed in Table 2
using groups M1 and Group 3 of V/Z/W. TABLE-US-00002 TABLE 2 1.1
1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9
3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4.1 4.2 4.3 4.4 4.5 4.6 4.7 4.8
4.9
[0463] Preferred compounds are also compounds of Tables 1 and 2 of
formulae VI-VIII where R.sup.7 is an L-valinyl group attached via a
carbonyl and R.sup.7 and R.sup.8 form a 5-membered cyclic
carbonate.
[0464] Moreover, the compounds of the present invention can be used
for inhibiting viral replication. In another aspect, the compounds
of this invention can be used for inhibiting RNA-dependent RNA
viral replication. In a further aspect, the compounds of this
invention can be used for inhibiting HCV replication.
[0465] In another aspect, the compounds of the present invention
can be used for treating viral infections. In a further aspect,
compounds of this invention can be used for treating RNA-dependent
RNA viral infection. In another aspect, compounds of this invention
can be used for treating HCV infection.
[0466] In another aspect, the compounds of the present invention
can be used for treating viral infections of the liver. In a
further aspect, compounds of this invention can be used for
treating RNA-dependent RNA viral infection in the liver. In another
aspect, compounds of this invention can be used for treating HCV
infection in the liver.
[0467] In one aspect, inhibition of viral replication is measured
in serum. Increased viral titer reduction is associated with
decreased generation of viral mutants which are associated with
drug resistance.
[0468] In another aspect, the compounds of the present invention
can be used for preventing the onset of symptoms associated with a
viral infection.
[0469] Activation of prodrugs of this invention results in the
production of a nucleoside monophosphate (NMP). NMPs are frequently
further phosphorylated inside the hepatocyte to the biologically
active nucleoside triphosphate (NTP). Drug elimination from the
hepatocyte typically entails degradation of phosphorylated
metabolites back to a species capable of being transported out of
the hepatocyte and into the blood for elimination by the kidney or
into the bile for biliary excretion. Often with nucleoside-based
drug the phophorylated metabolites are dephosphorylated to the
uncharged nucleoside.
[0470] Nucleosides that leak back into the systemic circulation
result in systemic exposure. If the nucleoside is active
systemically, e.g. through entry into virally infected cells and
phosphorylation to the active species, escape of the nucleoside
from the liver leads to biological activity outside of the liver
(i.e. extrahepatic tissues, blood cells). In this case, prodrugs of
the invention can be effective for treating diseases outside of the
liver, e.g. viral infections. Since many nucleosides exhibit poor
oral bioavailability due to breakdown in the gastrointestinal tract
either enzymatically (e.g. deamination by adenosine deaminase) or
chemically (e.g. acid instability), the prodrug can be used for
oral drug delivery. Moreover, given that the prodrugs in some cases
are broken down slowly relative to e.g. most ester based prodrugs,
the prodrugs could advantageously result in slow, sustained
systemic release of the nucleoside.
[0471] In other cases, however, systemic exposure to the nucleoside
can result in toxicity. This can be minimized by selecting
nucleosides that are preferentially excreted through the bile or
nucleosides that are unable to undergo phosphorylation in tissues
or nucleosides that undergo rapid intrahepatic metabolism to a
biologically inactive metabolite. Some enzymes in the hepatocyte
are present that can degrade nucleosides and therefore minimize
exposure (e.g. Phase I and Phase II enzymes). One example is
adenosine deaminase, which can deaminate some adenosine-based
nucleosides to produce the corresponding inosine analogue. Rapid
intracellular deamination of the nucleoside following its
dephosphorylation to the nucleoside limits systemic exposure to the
nucleoside and diminishes the risk of toxicity.
[0472] Methods described in Examples A-D were used to test
activation of compounds of this invention. Methods used in Example
E were used to evaluate the ability of compounds of the invention
to generate NTPs.
[0473] HCV replication in human liver tissue was evaluated as in
Example F. Liver specificity of the prodrugs relative to the
nucleosides was measured by methods in Example G.
[0474] Tissue distribution can be determined according to methods
in Example H. Oral bioavailability was determined by methods
described in Example I. The susceptibility of nucleoside analogs to
metabolism can be determined as in Example J.
[0475] In one aspect of the present invention, the RNA-dependent
RNA viral infection is a positive-sense single-stranded
RNA-dependent viral infection. In another aspect, the
positive-sense single-stranded RNA-dependent RNA viral infection is
Flaviviridae viral infection or Picornaviridae viral infection. In
a subclass of this class, the Picornaviridae viral infection is
rhinovirus infection, poliovirus infection, or hepatitis A virus
infection. In a second subclass of this class, the Flaviviridae
viral infection is selected from the group consisting of hepatitis
C virus infection, yellow fever virus infection, dengue virus
infection, West Nile virus infection, Japanese encephalitis virus
infection, Banzi virus infection, and bovine viral diarrhea virus
infection. In a subclass of this subclass, the Flaviviridae viral
infections hepatitis C virus infection.
[0476] In a further aspect, compounds of the present invention can
be used to enhance the oral bioavailability of the parent drug. In
another aspect, compounds of the present invention can be used to
enhance the oral bioavailability of the parent drug by at least 5%.
In another aspect, compounds of the present invention can be used
to enhance the oral bioavailability of the parent drug by at least
10%. In another aspect, oral bioavailability is enhanced by 50%
compared to the parent drug administered orally. In a further
aspect, the oral bioavailability is enhanced by at least 100%.
[0477] In another aspect, compounds of the present invention can be
used to increase the therapeutic index of a drug.
[0478] In one aspect, compounds of the present invention can be
used to bypass drug resistance.
[0479] In another aspect, compounds of the present invention can be
used to treat cancer.
Formulations
[0480] Compounds of the invention are administered in a total daily
dose of 0.01 to 1000 mg/kg. In one aspect the range is about 0.1
mg/kg to about 100 mg/kg. In another aspect the range is 0.5 to 20
mg/kg. The dose may be administered in as many divided doses as is
convenient.
[0481] Compounds of this invention when used in combination with
other antiviral agents may be administered as a daily dose or an
appropriate fraction of the daily dose (e.g., bid). Administration
of the prodrug may occur at or near the time in which the other
antiviral is administered or at a different time. The compounds of
this invention may be used in a multidrug regimen, also known as
combination or `cocktail` therapy, wherein, multiple agents may be
administered together, may be administered separately at the same
time or at different intervals, or administered sequentially. The
compounds of this invention may be administered after a course of
treatment by another agent, during a course of therapy with another
agent, administered as part of a therapeutic regimen, or may be
administered prior to therapy by another agent in a treatment
program.
[0482] For the purposes of this invention, the compounds may be
administered by a variety of means including orally, parenterally,
by inhalation spray, topically, or rectally in formulations
containing pharmaceutically acceptable carriers, adjuvants and
vehicles. The term parenteral as used here includes subcutaneous,
intravenous, intramuscular, and intraarterial injections with a
variety of infusion techniques. Intraarterial and intravenous
injection as used herein includes administration through catheters.
Intravenous administration is generally preferred.
[0483] Pharmaceutically acceptable salts include acetate, adipate,
besylate, bromide, camsylate, chloride, citrate, edisylate,
estolate, fumarate, gluceptate, gluconate, glucuronate, hippurate,
hyclate, hydrobromide, hydrochloride, iodide, isethionate, lactate,
lactobionate, maleate, mesylate, methylbromide, methylsulfate,
napsylate, nitrate, oleate, palmoate, phosphate, polygalacturonate,
stearate, succinate, sulfate, sulfosalicylate, tannate, tartrate,
terphthalate, tosylate, and triethiodide.
[0484] Pharmaceutical compositions containing the active ingredient
may be in any form suitable for the intended method of
administration. When used for oral use for example, tablets,
troches, lozenges, aqueous or oil suspensions, dispersible powders
or granules, emulsions, hard or soft capsules, syrups or elixirs
may be prepared. Compositions intended for oral use may be prepared
according to any method known to the art for the manufacture of
pharmaceutical compositions and such compositions may contain one
or more agents including sweetening agents, flavoring agents,
coloring agents and preserving agents, in order to provide a
palatable preparation. Tablets containing the active ingredient in
admixture with non-toxic pharmaceutically acceptable excipient
which are suitable for manufacture of tablets are acceptable. These
excipients may be, for example, inert diluents, such as calcium or
sodium carbonate, lactose, calcium or sodium phosphate; granulating
and disintegrating agents, such as maize starch, or alginic acid;
binding agents, such as starch, gelatin or acacia; and lubricating
agents, such as magnesium stearate, stearic acid or talc. Tablets
may be uncoated or may be coated by known techniques including
microencapsulation to delay disintegration and adsorption in the
gastrointestinal tract and thereby provide a sustained action over
a longer period. For example, a time delay material such as
glyceryl monostearate or glyceryl distearate alone or with a wax
may be employed.
[0485] Formulations for oral use may be also presented as hard
gelatin capsules where the active ingredient is mixed with an inert
solid diluent, for example calcium phosphate or kaolin, or as soft
gelatin capsules wherein the active ingredient is mixed with water
or an oil medium, such as peanut oil, liquid paraffin or olive
oil.
[0486] Aqueous suspensions of the invention contain the active
materials in admixture with excipients suitable for the manufacture
of aqueous suspensions. Such excipients include a suspending agent,
such as sodium carboxymethylcellulose, methylcellulose,
ethylcellulose, hydroxypropylcellulose, hydroxypropyl
methylcellulose, sodium alginate, polyvinylpyrrolidone, gum
tragacanth and gum acacia, and dispersing or wetting agents such as
a naturally occurring phosphatide (e.g., lecithin), a condensation
product of an alkylene oxide with a fatty acid (e.g.,
polyoxyethylene stearate), a condensation product of ethylene oxide
with a long chain aliphatic alcohol (e.g.,
heptadecaethyleneoxycetanol), a condensation product of ethylene
oxide with a partial ester derived from a fatty acid and a hexitol
anhydride (e.g., polyoxyethylene sorbitan monooleate). The aqueous
suspension may also contain one or more preservatives such as ethyl
or n-propyl p-hydroxy-benzoate, one or more coloring agents, one or
more flavoring agents and one or more sweetening agents, such as
sucrose or saccharin.
[0487] Oil suspensions may be formulated by suspending the active
ingredient in a vegetable oil, such as arachid oil, olive oil,
sesame oil or coconut oil, or in a mineral oil such as liquid
paraffin. The oral suspensions may contain a thickening agent, such
as beeswax, hard paraffin or cetyl alcohol. Sweetening agents, such
as those set forth above, and flavoring agents may be added to
provide a palatable oral preparation. These compositions may be
preserved by the addition of an antioxidant such as ascorbic
acid.
[0488] Dispersible powders and granules of the invention suitable
for preparation of an aqueous suspension by the addition of water
provide the active ingredient in admixture with a dispersing or
wetting agent, a suspending agent, and one or more preservatives.
Suitable dispersing or wetting agents and suspending agents are
exemplified by those disclosed above. Additional excipients, for
example sweetening, flavoring and coloring agents, may also be
present.
[0489] The pharmaceutical compositions of the invention may also be
in the form of oil-in-water emulsions. The oily phase may be a
vegetable oil, such as olive oil or arachid oil, a mineral oil,
such as liquid paraffin, or a mixture of these. Suitable
emulsifying agents include naturally-occurring gums, such as gum
acacia and gum tragacanth, naturally occurring phosphatides, such
as soybean lecithin, esters or partial esters derived from fatty
acids and hexitol anhydrides, such as sorbitan monooleate, and
condensation products of these partial esters with ethylene oxide,
such as polyoxyethylene sorbitan monooleate. The emulsion may also
contain sweetening and flavoring agents.
[0490] Syrups and elixirs may be formulated with sweetening agents,
such as glycerol, sorbitol or sucrose. Such formulations may also
contain a demulcent, a preservative, a flavoring or a coloring
agent.
[0491] The pharmaceutical compositions of the invention may be in
the form of a sterile injectable preparation, such as a sterile
injectable aqueous or oleaginous suspension. This suspension may be
formulated according to the known art using those suitable
dispersing or wetting agents and suspending agents which have been
mentioned above. The sterile injectable preparation may also be a
sterile injectable solution or suspension in a non-toxic
parenterally acceptable diluent or solvent, such as a solution in
1,3-butane-diol or prepared as a lyophilized powder. Among the
acceptable vehicles and solvents that may be employed are water,
Ringer's solution and isotonic sodium chloride solution. In
addition, sterile fixed oils may conventionally be employed as a
solvent or suspending medium. For this purpose any bland fixed oil
may be employed including synthetic mono- or diglycerides. In
addition, fatty acids such as oleic acid may likewise be used in
the preparation of injectables.
[0492] The amount of active ingredient that may be combined with
the carrier material to produce a single dosage form will vary
depending upon the host treated and the particular mode of
administration. For example, a time-release formulation intended
for oral administration to humans may contain 20 to 2000 .mu.mol
(approximately 10 to 1000 mg) of active material compounded with an
appropriate and convenient amount of carrier material which may
vary from about 5 to about 95% of the total compositions. It is
preferred that the pharmaceutical composition be prepared which
provides easily measurable amounts for administration. For example,
an aqueous solution intended for intravenous infusion should
contain from about 0.05 to about 50 .mu.mol (approximately 0.025 to
25 mg) of the active ingredient per milliliter of solution in order
that infusion of a suitable volume at a rate of about 30 mL/h can
occur.
[0493] As noted above, formulations of the present invention
suitable for oral administration may be presented as discrete units
such as capsules, cachets or tablets each containing a
predetermined amount of the active ingredient; as a powder or
granules; as a solution or a suspension in an aqueous or
non-aqueous liquid; or as an oil-in-water liquid emulsion or a
water-in-oil liquid emulsion. The active ingredient may also be
administered as a bolus, electuary or paste.
[0494] A tablet may be made by compression or molding, optionally
with one or more accessory ingredients. Compressed tablets may be
prepared by compressing in a suitable machine the active ingredient
in a free flowing form such as a powder or granules, optionally
mixed with a binder (e.g., povidone, gelatin, hydroxypropylmethyl
cellulose), lubricant, inert diluent, preservative, disintegrant
(e.g., sodium starch glycolate, cross-linked povidone, cross-linked
sodium carboxymethyl cellulose) surface active or dispersing agent.
Molded tablets may be made by molding in a suitable machine a
mixture of the powdered compound moistened with an inert liquid
diluent. The tablets may optionally be coated or scored and may be
formulated so as to provide slow or controlled release of the
active ingredient therein using, for example, hydroxypropyl
methylcellulose in varying proportions to provide the desired
release profile. Tablets may optionally be provided with an enteric
coating, to provide release in parts of the gut other than the
stomach. This is particularly advantageous with the compounds of
Formula I when such compounds are susceptible to acid
hydrolysis.
[0495] Formulations suitable for topical administration in the
mouth include lozenges comprising the active ingredient in a
flavored base, usually sucrose and acacia or tragacanth; pastilles
comprising the active ingredient in an inert base such as gelatin
and glycerin, or sucrose and acacia; and mouthwashes comprising the
active ingredient in a suitable liquid carrier.
[0496] Formulations for rectal administration may be presented as a
suppository with a suitable base comprising for example cocoa
butter or a salicylate.
[0497] Formulations suitable for vaginal administration may be
presented as pessaries, tampons, creams, gels, pastes, foams or
spray formulations containing in addition to the active ingredient
such carriers as are known in the art to be appropriate.
[0498] Formulations suitable for parenteral administration include
aqueous and non-aqueous isotonic sterile injection solutions which
may contain antioxidants, buffers, bacteriostats and solutes which
render the formulation isotonic with the blood of the intended
recipient; and aqueous and non-aqueous sterile suspensions which
may include suspending agents and thickening agents. The
formulations may be presented in unit-dose or multi-dose sealed
containers, for example, ampoules and vials, and may be stored in a
freeze-dried (lyophilized) condition requiring only the addition of
the sterile liquid carrier, for example water for injections,
immediately prior to use. Injection solutions and suspensions may
be prepared from sterile powders, granules and tablets of the kind
previously described.
[0499] Formulations suitable for parenteral administration may be
administered in a continuous infusion manner via an indwelling pump
or via a hospital bag. Continuous infusion includes the infusion by
an external pump. The infusions may be done through a Hickman or
PICC or any other suitable means of administering a formulation
either parenterally or i.v.
[0500] Preferred unit dosage formulations are those containing a
daily dose or unit, daily sub-dose, or an appropriate fraction
thereof, of a drug.
[0501] It will be understood, however, that the specific dose level
for any particular patient will depend on a variety of factors
including the activity of the specific compound employed; the age,
body weight, general health, sex and diet of the individual being
treated; the time and route of administration; the rate of
excretion; other drugs which have previously been administered; and
the severity of the particular disease undergoing therapy, as is
well understood by those skilled in the art.
[0502] Another aspect of the present invention is concerned with a
method of inhibiting HCV replication or treating HCV infection with
a compound of the present invention in combination with one or more
agents useful for treating HCV infection. Such agents active
against HCV include, but are not limited to, ribavirin, levovirin,
viramidine, thymosin alpha-1, interferon-.beta.,
interferon-.alpha., pegylated interferon-.alpha.
(peginterferon-.alpha.), a combination of interferon-.alpha. and
ribavirin, a combination of peginterferon-.alpha. and ribavirin, a
combination of interferon-.alpha. and levovirin, and a combination
of peginterferon-.alpha. and levovirin. Interferon-.alpha.
includes, but is not limited to, recombinant interferon-.alpha.2a
(such as Roferon interferon available from Hoffmann-LaRoche,
Nutley, N.J.), pegylated interferon-.alpha.2a (Pegasys.TM.),
interferon-.alpha.2b (such as Intron-A interferon available from
Schering Corp., Kenilworth, N.J.), pegylated interferon-.alpha.2b
(PegIntron.TM.), a recombinant consensus interferon (such as
interferon alphacon-1), and a purified interferon-.alpha. product.
Amgen's recombinant consensus interferon has the brand name
Infergen.RTM.. Levovirin is the L-enantiomer of ribavirin which has
shown immunomodulatory activity similar to ribavirin. Viramidine is
a liver-targeting prodrug analog of ribavirin disclosed in WO
01/60379 (assigned to ICN Pharmaceuticals). In accordance with this
method of the present invention, the individual components of the
combination can be administered separately at different times
during the course of therapy or concurrently in divided or single
combination forms. The instant invention is therefore to be
understood as embracing all such regimes of simultaneous or
alternating treatment, and the term "administering" is to be
interpreted accordingly. It will be understood that the scope of
combinations of the compounds of this invention with other agents
useful for treating HCV infection includes in principle any
combination with any pharmaceutical composition for treating HCV
infection. When a compound of the present invention or a
pharmaceutically acceptable salt thereof is used in combination
with a second therapeutic agent active against HCV, the dose of
each compound may be either the same as or different from the dose
when the compound is used alone.
[0503] Also included within the scope of the invention is a
pharmaceutical composition comprising a compound of Formula I or
prodrug or pharmaceutically acceptable salt thereof and at least
one agent useful for treating a viral infection, particularly an
HCV infection.
[0504] For the treatment of HCV infection, the compounds of the
present invention may also be administered in combination with an
agent that is an inhibitor of HCV NS3 serine protease. HCV NS3
serine protease is an essential viral enzyme and has been described
to be an excellent target for inhibition of HCV replication. Both
substrate and non-substrate based inhibitors of HCV NS3 protease
inhibitors are disclosed in WO 98/22496, WO 98/46630, WO 99/07733,
WO 99/07734, WO 99/38888, WO 99/50230, WO 99/64442, WO 00/09543, WO
00/59929, GB-2337262, WO 02/48116, WO 02/48172, U.S. Pat. No.
6,323,180, and U.S. Pat. No. 6,410,531. Specific embodiments of NS3
protease inhibitors for combination with the compounds of the
present invention are BILN 2061 (Boehringer Ingelheim) and
VX-950/LY-570310. HCV NS3 protease as a target for the development
of inhibitors of HCV replication and for the treatment of HCV
infection is discussed in B. W. Dymock, "Emerging therapies for
hepatitis C virus infection," Emerging Drugs, 6: 13-42 (2001).
[0505] Ribavirin, levovirin, and viramidine may exert their
anti-HCV effects by modulating intracellular pools of guanine
nucleotides via inhibition of the intracellular enzyme inosine
monophosphate dehydrogenase (IMPDH). IMPDH is the rate-limiting
enzyme on the biosynthetic route in de novo guanine nucleotide
biosynthesis. Ribavirin is readily phosphorylated intracellularly
and the monophosphate derivative is an inhibitor of IMPDH. Thus,
inhibition of IMPDH represents another useful target for the
discovery of inhibitors of HCV replication. Therefore, the
compounds of the present invention may also be administered in
combination with an inhibitor of IMPDH, such as VX-497
(merimepodib), which is disclosed in WO 97/41211 and WO 01/00622
(assigned to Vertex); another IMPDH inhibitor, such as that
disclosed in WO 00/25780 (assigned to Bristol-Myers Squibb); or
mycophenolate mofetil [see A. C. Allison and E. M. Eugui, Agents
Action, 44 (Suppl.): 165 (1993)].
[0506] For the treatment of HCV infection, the compounds of the
present invention may also be administered in combination with the
antiviral agent amantadine (1-aminoadamantane) and its
hydrochloride salt [for a comprehensive description of this agent,
see J. Kirschbaum, Anal. Profiles Drug Subs. 12: 1-36 (1983)].
[0507] The compounds of the present invention may also be combined
for the treatment of HCV infection with antiviral 1'-C, 2'-C-, or
3'-C-branched ribonucleosides disclosed in R. E. Harry-O'kuru, et
al., J. Org. Chem., 62: 1754-1759 (1997); M. S. Wolfe, et al.,
Tetrahedron Lett., 36: 7611-7614 (1995); U.S. Pat. No. 3,480,613
(Nov. 25, 1969); International Publication Number WO 01/90121 (29
Nov. 2001); International Publication Number WO 01/92282 (6 Dec.
2001); and International Publication Number WO 02/32920 (25 Apr.
2002); the contents of each of which are incorporated by reference
in their entirety. Such branched ribonucleosides include, but are
not limited to, 2'-C-methylcytidine, 2'-C-methyluridine,
2'-C-methyladenosine, 2'-C-methylguanosine, and
9-(2-C-methyl-.beta.-D-ribofuranosyl)-2,6-diaminopurine, and
prodrugs thereof.
[0508] The compounds of the present invention may also be combined
for the treatment of HCV infection with other nucleosides having
anti-HCV properties, such as those disclosed in WO 02/51425 (4 Jul.
2002), assigned to Mitsubishi Pharma Corp.; WO 01/79246, WO
02/32920 (25 Apr. 2002), and WO 02/48165 (20 Jun. 2002), assigned
to Pharmasset, Ltd.; WO 01/68663 (20 Sep. 2001), assigned to ICN
Pharmaceuticals; WO 99/43691 (2 Sep. 1999); WO 02/18404 (7 Mar.
2002), assigned to Hoffmann-LaRoche; U.S. 2002/0019363 (14 Feb.
2002); WO 02/057287 (25 Jul. 2002), assigned to Merck & Co. and
Isis Pharmaceuticals; and WO 02/057425 (25 Jul. 2002), assigned to
Merck & Co. and Isis Pharmaceuticals.
[0509] The compounds of the present invention may also be combined
for the treatment of HCV infection with non-nucleoside inhibitors
of HCV polymerase such as those disclosed in WO 01/77091 (18 Oct.
2001), assigned to Tularik, Inc.; WO 01/47883 (5 Jul. 2001),
assigned to Japan Tobacco, Inc.; WO 02/04425 (17 Jan. 2002),
assigned to Boehringer Ingelheim; WO 02/06246 (24 Jan. 2002),
assigned to Istituto di Ricerche di Biologia Moleculare P.
Angeletti S. P. A.; and WO 02/20497 (3 Mar. 2002). WO 01/47883
discloses a large number of benzimidazole derivatives, such as
JTK-003, which is claimed to be an orally active inhibitor of NS5B
that is currently undergoing clinical evaluation.
Synthesis of Compounds of 2'-C-Methyl Derivatives
[0510] Synthesis of the 5'-nucleoside monophosphate (NMP) prodrugs
of the present invention is organized into two sections: 1.
synthesis of phosphorylation precursors; 2. synthesis of prodrugs
via coupling of nucleosides and prodrug moiety.
Synthesis of Phosphorylation Precursors:
[0511] Synthesis of phosphorylation precursors is attained in two
stages: 1. Synthesis of 1,3-diols and 2. Synthesis of
phosphorylation precursor.
Synthesis of 1,3-Diols:
[0512] A variety of synthetic methods are known to prepare the
following types of 1,3-diols: a) 1-substituted; b) 2-substituted;
and c) 1,2- or 1,3-annulated in their racemic or enantioenriched
form. The V, W, Z groups of Formula I can be introduced or modified
either during synthesis of diols or after the synthesis of
prodrugs.
Synthesis of 1-(aryl)-Propane-1,3-Diols:
[0513] The suitable methods to prepare 1,3-diols are divided into
two types as following: 1) synthesis of racemic
1-(aryl)-propane-1,3-diols; 2) synthesis of enantioenriched
1-(aryl)-propane-1,3-diols.
Synthesis of Racemic 1-(aryl)-Propane-1,3-Diol:
[0514] 1,3-Dihydroxy compounds can be synthesized by several
well-known methods from the literature. Substituted aromatic
aldehydes are utilized to synthesize racemic
1-(aryl)propane-1,3-diols via addition of lithium enolate of alkyl
acetate followed by ester reduction (path A) (Turner, J. Org. Chem.
55:4744 (1990)). Alternatively, aryl lithium or aryl Grignard
additions to 1-hydroxy propan-3-al also give
1-(arylsubstituted)propane-1,3-diols (path B). This method will
enable conversion of various substituted aryl halides to
1-(arylsubstituted)-1,3-propane diols (Coppi, et al., J. Org. Chem.
53:911 (1988)). Aryl halides can also be used to synthesize
1-substituted propane diols by Heck coupling of 1,3-diox-4-ene
followed by reduction and hydrolysis (Sakamoto, et al., Tetrahedron
Lett. 33:6845 (1992)). Pyridyl-, quinolyl-, isoquinolyl-propan-3-ol
derivatives can be hydroxylated to 1-substituted-1,3-diols by
N-oxide formation followed by rearrangement in the presence of
acetic anhydride (path C) (Yamamoto, et al., Tetrahedron 37:1871
(1981)). A variety of aromatic aldehydes can also be converted to
1-substituted-1,3-diols by vinyl lithium or vinyl Grignard addition
followed by hydroboration reaction (path D). ##STR55## Synthesis of
Enantioenriched 1-(aryl)-Propane-1,3-Diol:
[0515] A variety of known methods for resolution of secondary
alcohols via chemical or enzymatic agents may be utilized for
preparation of diol enantiomers (Harada, et al., Tetrahedron Lett.
28:4843 (1987)). Transition metal catalyzed hydrogenation of
substituted 3-aryl-3-oxo-propionic acids or esters is an efficient
method to prepare R- or S-isomers of beta hydroxy acids or esters
in high enantiomeric purity (Comprehensive Asymmetric Catalysis,
Jacobsen, E. N., Pfaltz, A., Yamamoto, H. (Eds), Springer, (1999);
Asymmetric Catalysis in Organic Synthesis, Noyori, R., John Wiley,
(1994)). These beta hydroxy acid or ester products can be further
reduced to give required 1-(aryl)-propane-1,3-diols in high
enantiomeric excess (ee). (path A). The .beta.-keto acid or ester
substrates for high pressure hydrogenation or hydrogen transfer
reactions may be prepared by a variety of methods such as
condensation of acetophenone with dimethylcarbonate in the presence
of a base (Chu, et al., J. Het Chem. 22:1033 (1985)), by ester
condensation (Turner, et al., J. Org. Chem. 54:4229 (1989)) or from
aryl halides (Kobayashi, et al., Tetrahedron Lett. 27:4745 (1986)).
Alternatively, 1,3-diols of high enantiomeric purity can be
obtained by enantioselective borane reduction of
.beta.-hydroxyethyl aryl ketone derivatives or .beta.-keto acid
derivatives (path B) (Ramachandran, et al., Tetrahedron Lett.
38:761 (1997)). In another method, commercially available cinnamyl
alcohols may be converted to epoxy alcohols under catalytic
asymmetric epoxidation conditions. These epoxy alcohols are reduced
by Red-Al to result in 1,3-diols with high ee's (path C) (Gao, et
al., J. Org. Chem. 53:4081 (1980)). Enantioselective aldol
condensation is another well-described method for synthesis of
1,3-oxygenated functionality with high ee's starting from aromatic
aldehydes. (path D) (Mukaiyama, Org. React. 28:203 (1982)).
##STR56## Synthesis of 2-Substituted 1,3-Diols:
[0516] Various 2-substituted-1,3-diols can be made from
commercially available 2-(hydroxymethyl)-1,3-propane-diol.
Pentaerythritol can be converted to triol via decarboxylation of
diacid followed by reduction (path a) (Werle, et al., Liebigs. Ann.
Chem., 1986, 944) or diol-monocarboxylic acid derivatives can also
be obtained by decarboxylation under known conditions (Iwata, et.
al., Tetrahedron Lett. 1987, 28, 3131). Nitrotriol is also known to
give triol by reductive elimination (path b) (Latour, et. al.,
Synthesis, 1987, 8, 742). The triol can be derivatized by mono
acylation or carbonate formation by treatment with alkanoyl
chloride, or alkylchloroformate (path d) (Greene and Wuts,
Protective groups in organic synthesis, John Wiley, New York,
1990). Aryl substitution can be affected by oxidation to aldehyde
and aryl Grignard additions (path c). Aldehydes can also be
converted to substituted amines by reductive animation reaction
(path e). ##STR57## Synthesis of cyclic-1,3-diols:
[0517] Compounds of Formula 1 where V-Z or V-W are fused by four
carbons are made from cyclohexane diol derivatives. Commercially
available cis, cis-1,3,5-cyclohexane-triol can be used as is or
modified as described in case of 2-substituted propan-1,3-diols to
give various analogues. These modifications can either be made
before or after ester formation. Various 1,3-cyclohexane-diols can
be made by Diels-Alder methodology using pyrone as diene (Posner,
et. al., Tetrahedron Lett., 1991, 32, 5295). Cyclohexanediol
derivatives are also made by nitrile oxide-olefin additions
(Curran, et. al., J. Am. Chem. Soc., 1985, 107, 6023).
Alternatively, cyclohexyl precursors are also made from
commercially available quinic acid (Rao, et. al., Tetrahedron
Lett., 1991, 32, 547.)
Synthesis of Substituted 1,3-hydroxyamines and 1,3-diamines:
[0518] A large number of synthetic methods are available for the
preparation of substituted 1,3-hydroxyamines and 1,3-diamines due
to the ubiquitous nature of these functionalities in naturally
occurring compounds. Following are some of these methods organized
into: 1. synthesis of substituted 1,3-hydroxy amines; 2. synthesis
of substituted 1,3-diamines and 3. synthesis of chiral substituted
1,3-hydroxyamines and 1,3-diamines.
Synthesis of Substituted 1,3-hydroxy Amines:
[0519] 1,3-Diols described in the earlier section can be converted
selectively to either hydroxy amines or to corresponding diamines
by converting hydroxy functionality to a leaving group and treating
with anhydrous ammonia or required primary or secondary amines
(Corey, et al., Tetrahedron Lett., 1989, 30, 5207: Gao, et al., J.
Org. Chem., 1988, 53, 4081). A similar transformation may also be
achieved directly from alcohols under Mitsunobu type of reaction
conditions (Hughes, D. L., Org. React., 1992, 42).
[0520] A general synthetic procedure for
3-aryl-3-hydroxy-propan-1-amine type of prodrug moiety involves
aldol type condensation of aryl esters with alkyl nitrites followed
by reduction of resulting substituted benzoylacetonitrile (Shih et
al., Heterocycles, 1986, 24, 1599). The procedure can also be
adapted for formation of 2-substituted aminopropanols by using
substituted alkylnitrile. In another approach,
3-aryl-3-amino-propan-1-ol type of prodrug groups are synthesized
from aryl aldehydes by condensation of malonic acid in presence of
ammonium acetate followed by reduction of resulting substituted
.beta.-amino acids. Both these methods enable to introduce wide
variety of substitution of aryl group (Shih, et al., Heterocycles.,
1978, 9, 1277). In an alternate approach, .beta.-substituted
organolithium compounds of 1-amino-1-aryl ethyl dianion generated
from styrene type of compounds undergo addition with carbonyl
compounds to give variety of W, W' substitution by variation of the
carbonyl compounds (Barluenga, et al., J. Org. Chem., 1979, 44,
4798).
Synthesis of Substituted 1,3-diamines:
[0521] Substituted 1,3-diamines are synthesized starting from a
variety of substrates. Arylglutaronitriles can be transformed to
1-substituted diamines by hydrolysis to amide and Hofmann
rearrangement conditions (Bertochio, et al., Bull. Soc. Chim. Fr,
1962, 1809). Whereas, malononitrile substitution will enable
variety of Z substitution by electrophile introduction followed by
hydride reduction to corresponding diamines. In another approach,
cinnamaldehydes react with hydrazines or substituted hydrazines to
give corresponding pyrazolines which upon catalytic hydrogenation
result in substituted 1,3-diamines (Weinhardt, et al., J. Med.
Chem., 1985, 28, 694). High trans-diastereoselectivity of
1,3-substitution is also attainable by aryl Grignard addition on to
pyrazolines followed by reduction (Alexakis, et al., J. Org. Chem.,
1992, 576, 4563). 1-Aryl-1,3-diaminopropanes are also prepared by
diborane reduction of 3-amino-3-arylacrylonitriles which in turn
are made from nitrile substituted aromatic compounds (Dornow, et
al., Chem Ber., 1949, 82, 254). Reduction of 1,3-diimines obtained
from corresponding 1,3-carbonyl compounds are another source of
1,3-diamine prodrug moiety which allows a wide variety of
activating groups V and/or Z (Barluenga, et al., J. Org. Chem.,
1983, 48, 2255).
Synthesis of Chiral Substituted 1,3-hydroxyamines and
1,3-diamines:
[0522] Enantiomerically pure 3-aryl-3-hydroxypropan-1-amines are
synthesized by CBS enantioselective catalytic reaction of
.beta.-chloropropiophenone followed by displacement of halo group
to make secondary or primary amines as required (Corey, et al.,
Tetrahedron Lett., 1989, 30, 5207). Chiral 3-aryl-3-amino
propan-1-ol type of prodrug moiety may be obtained by 1,3-dipolar
addition of chirally pure olefin and substituted nitrone of
arylaldehyde followed by reduction of resulting isoxazolidine
(Koizumi, et al., J. Org. Chem., 1982, 47, 4005). Chiral induction
in 1,3-polar additions to form substituted isoxazolidines is also
attained by chiral phosphine palladium complexes resulting in
enantioselective formation of amino alcohols (Hori, et al., J. Org.
Chem., 1999, 64, 5017). Alternatively, optically pure 1-aryl
substituted amino alcohols are obtained by selective ring opening
of corresponding chiral epoxy alcohols with desired amines (Canas
et al., Tetrahedron Lett., 1991, 32, 6931).
[0523] Several methods are known for diastereoselective synthesis
of 1,3-disubstituted aminoalcohols. For example, treatment of
(E)-N-cinnamyltrichloroacetamide with hypochlorous acid results in
trans-dihydrooxazine which is readily hydrolysed to
erythro-.beta.-chloro-.gamma.-hydroxy-.gamma.-phenylpropanamine in
high diastereoselectivity (Commercon et al., Tetrahedron Lett.,
1990, 31, 3871). Diastereoselective formation of 1,3-aminoalcohols
is also achieved by reductive amination of optically pure 3-hydroxy
ketones (Haddad et al., Tetrahedron Lett., 1997, 38, 5981). In an
alternate approach, 3-aminoketones are transformed to
1,3-disubstituted aminoalcohols in high stereoselectivity by a
selective hydride reduction (Barluenga et al., J. Org. Chem., 1992,
57, 1219).
Synthesis of Phosphorylation Precursors:
[0524] Synthesis of phosphorylation precursors is divided in to two
sections: a. synthesis of P(III) phosphorylation precursor, b.
stereoselective synthesis of P(V) phosphorylation precursors.
Synthesis of P(III) Phosphorylation Precursors: ##STR58##
[0525] Phosphorylation of 5'-alcohol is achieved using cyclic
1',3'-propanyl esters of phosphorylating agents where the agent is
in the P(III) oxidation state. One preferred phosphorylating agent
is a chloro phospholane (L'=chloro). Cyclic chlorophospholanes are
prepared under mild conditions by reaction of phosphorus
trichloride with substituted 1,3-diols (Wissner, et al, J. Med.
Chem., 1992, 35, 1650). Alternatively phosphoramidites can be used
as the phosphorylating agent (Beaucage, et al., Tetrahedron, 1993,
49, 6123). Appropriately substituted phosphoramidites can be
prepared by reacting cyclic chlorophospholanes with
N,N-dialkylamine (Perich, et al., Aust. J. Chem., 1990, 43, 1623.
Perich, et al, Synthesis, 1988, 2, 142) or by reaction of
commercially available dialkylaminophosphorochloridate with
substituted propane-1,3-diols. Synthesis of P(V) Phosphorylation
Precursors: ##STR59##
[0526] In general, synthesis of phosphate esters is achieved by
coupling the alcohol with the corresponding activated phosphate
precursor for example, Chlorophosphate (L'=chloro) condensation
with 5'-hydroxy of nucleoside is a well known method for
preparation of nucleoside phosphate monoesters. The activated
precursor can be prepared by several well known methods.
Chlorophosphates useful for synthesis of the prodrugs are prepared
from the substituted-1,3-propanediol (Wissner, et al, J. Med.
Chem., 1992, 35, 1650). Chlorophosphates are made by oxidation of
the corresponding chlorophospholanes (Anderson, et al, J. Org.
Chem., 1984, 49, 1304), which are obtained by reaction of the
substituted diol with phosphorus trichloride. Alternatively, the
chlorophosphate agent is made by treating substituted-1,3-diols
with phosphorus oxychloride (Patois, et al, J. Chem. Soc. Perkin
Trans. I, 1990, 1577). Chlorophosphate species may also be
generated in situ from corresponding cyclic phosphites (Silverburg,
et al., Tetrahedron Lett., 1996, 37, 771), which in turn can be
either made from a chlorophospholane or phosphoramidate
intermediate. Phosphorofluoridate intermediate prepared either from
pyrophosphate or phosphoric acid may also act as precursor in
preparation of cyclic prodrugs (Watanabe et al., Tetrahedron Lett.,
1988, 29, 5763).
[0527] Phosphoramidates (L'=NRR') are also well-known intermediates
for the synthesis of phosphate esters. Monoalkyl or
dialkylphosphoramidate (Watanabe, et al, Chem Pharm Bull., 1990,
38, 562), triazolophosphoramidate (Yamakage, et al., Tetrahedron,
1989, 45, 5459) and pyrrolidinophosphoramidate (Nakayama, et al, J.
Am. Chem. Soc., 1990, 112, 6936) are some of the known
intermediates used for the preparation of phosphate esters. Another
effective phosphorylating procedure is a metal catalyzed addition
of cyclic chlorophosphate adduct of 2-oxazolone. This intermediate
attains high selectivity in phosphorylation of primary hydroxy
group in presence of secondary hydroxyl group (Nagamatsu, et al,
Tetrahedron Lett., 1987, 28, 2375). These agents are obtained by
reaction of a chlorophosphate with the amine or alternatively by
formation of the corresponding phosphoramidite followed by
oxidation. Synthesis of Enantiomerically Enriched P(V)
Phosphorylation Precursors: ##STR60##
[0528] The enantioenriched activated phosphorylating agent is
synthesized by phosphorylation of an enantioenriched
1-(V)-1,3-propanediol with phosphorodichloridates of formula
L--P(O)Cl.sub.2 in the presence of a base (Ferroni, et al., J. Org.
Chem. 64(13), 4943 (1999)). Phosphorylation of an enantiomerically
pure substituted diol with, for example, a commercially available
phosphorodichloridate R--OP(O)Cl.sub.2, where RO is a leaving
group, preferably aryl substituted with electron withdrawing
groups, such as a nitro or a chloro, produces two diastereomeric
intermediates. The relative configuration of the phosphorus atom is
easily determined by comparison of the .sup.31P NMR spectra. The
chemical shift of the equatorial phosphoryloxy moiety
(trans-isomer) is always more upfield than the one of the axial
isomer (cis-isomer) (Verkade, et al, J. Org. Chem., 1977, 42,
1549). These diastereomers can be further equilibrated to give a
trans-2,4-substituted phosphorylating agents in presence of a base
such as triethyl amine or DBU. The equilibration to complete
inversion of 2,4-cis-diastereomer is also achieved in presence of
appropriately substituted sodium phenoxide. The equilibration step
results in greater than 95% ee of the isolated
trans-phosphorylating agent.
Synthesis of Nucleosides.
[0529] All nucleoside moieties of Formula I are well described in
the literature. 2'-C-methyl-adenosine and 2'-C-methyl-guanosine
analogs are made by Lewis acid catalyzed reactions of the
persilylated base and 1'-acetate or benzoate sugar intermediate
(Walton et al., J. Am. Chem. Soc., 1966, 88, 4524; Harry-O'Kuru et
al., J. Org. Chem., 1997, 62, 1754, WO01/90121). The 7-deaza
analogs are made as described earlier from 1'-bromo sugar
intermediate via reaction of sodium salt of the bases
(US2002-0147160A1 or WO02/057827). The glycosylation products are
subjected to deprotection and amination via ammonolysis
reaction.
[0530] The nucleoside moieties and derivatives thereof of Formulae
VI-VIII of the present invention may be synthesized by many
well-established general methods described in the nucleoside
literature. Several nucleosides analogs described herein are
synthesized as illustrated in WO04/046331 and by the methods cited
therein. These compounds of the present invention can also be made
from a wide variety of commercial bases utilizing the 2'-methyl
riboglycosylation precursor (US2002-0147160A1 or WO02/057827) via a
range of well-known glycosylation reactions (Vorbruggen and
Ruh-Pohlenz, Handbook of Nucleoside Synthesis, Wiley, New York,
2001). Furthermore, deaza and aza nucleoside analogs may be
prepared utilizing the methods reported in the case of
corresponding ribo-analogs by glycosylation with 2'-methyl
glycosylation precursor (Robins, et al., Advances in Antiviral Drug
Design, Vol. 1, p 39-85, De Clercq, ed., JAI Press, Greenwich,
Conn., 1993). In addition, new base analogs of the nucleosides can
be synthesized by modification of the available nucleosides or via
synthesis of new bases followed by glycosylation (Chemistry of
Nucleosides and Nucleotides, Vols. 1-3, Townsend, ed., Plenum, New
York, 1988 and Nucleic Acid Chemistry, Vols. 1-4, Townsend and
Tipson Eds., Wiley, New York, 1986).
Synthesis of Prodrugs Via Coupling of Nucleosides and Prodrug
Moiety.
[0531] The following procedures on the preparation of prodrugs
illustrate the general procedures used to prepare the NMP prodrugs.
Prodrugs can be introduced at different stages of the synthesis.
Most often they are made at a later stage, because of the general
sensitivity of these groups to various reaction conditions.
Optically pure prodrugs containing single isomer at phosphorus
center are made by coupling of enantiomerically enriched activated
phosphate intermediates.
[0532] All the procedures described herein, where Y and Y' are
oxygen are also applicable for the preparation of the prodrugs
where Y and and/or Y' are NH by appropriate substitution or
protection of nitrogen.
[0533] The preparation of prodrugs is further organized into, 1)
synthesis via P(III) intermediates, 2) synthesis via P(V)
intermediates, and 3) miscellaneous methods. Synthesis of Prodrugs
Via P(III) Intermediates: ##STR61## wherein Q is N or CH; and L is
H and M is NH.sub.2 or M is OH and L is NH.sub.2.
[0534] Chlorophospholanes are used to phosphorylate alcohols on
nucleosides in the presence of an organic base (e.g.,
triethylamine, pyridine). Alternatively, the phosphite can be
obtained by coupling the nucleoside with a phosphoramidate in the
presence of a coupling promoter such as tetrazole or
benzimidazolium triflate (Hayakawa et al., J. Org. Chem., 1996, 61,
7996). Phosphite diastereomers may be isolated by column
chromatography or crystallization (Wang, et al, Tetrahedron Lett,
1997, 38, 3797; Bentridge et al., J. Am. Chem. Soc., 1989, 111,
3981). Since condensation of alcohols with chlorophospholanes or
phosphoramidites is an S.sub.N2(P) reaction, the product is
expected to have an inverted configuration. This allows for the
stereoselective synthesis of cyclic phosphites. Isomeric mixtures
of phosphorylation reactions can also be equilibrated (e.g. thermal
equilibration) to a more thermodynamically stable isomer.
[0535] The resulting phosphites are subsequently oxidized to the
corresponding phosphate prodrugs using an oxidant such as molecular
oxygen or t-butylhydroperoxide (Meier et al., Bioorg, Med. Chem.
Lett., 1997, 7, 1577). Oxidation of optically pure phosphites is
expected to stereoselectively provide optically active prodrugs
(Mikolajczyk, et al., J. Org. Chem., 1978, 43, 2132. Cullis, P. M.
J. Chem. Soc., Chem Commun., 1984, 1510, Verfurth, et al., Chem.
Ber., 1991, 129, 1627).
Synthesis of Prodrugs Via P(V) Intermediates:
[0536] For the synthesis of cis-prodrugs of Formula I, the prodrug
moiety can be introduced at different stages of the synthesis. Most
often the cyclic phosphates are introduced at a later stage,
because of the general sensitivity of these groups to various
reaction conditions. The synthesis can also proceed through using a
protected or unprotected nucleoside or nucleoside analog depending
on the reactivity of the functional groups present in the compound.
Single stereoisomers of the cis-prodrugs can be made either by
separation of the diastereoisomers/enantiomers by a combination of
column chromatography and/or crystallization, or by enantiospecific
or enantioselective synthesis using enantioenriched activated
phosphate intermediates. Synthesis of Enantiomerically Enriched
Prodrugs: ##STR62## wherein Q is N or CH; and L is H and M is
NH.sub.2 or M is OH and L is NH.sub.2.
[0537] The general procedure for the phosphorylation of protected
nucleosides is accomplished by reacting a suitably protected
nucleoside with a base and reacting the alkoxide generated with the
phosphorylating reagent. The protected nucleoside can be prepared
by one skilled in the art using one of the many procedures
described for the protection of nucleosides (Greene T. W.,
Protective Groups in Organic Chemistry, John Wiley & Sons, New
York (1999)). The nucleoside is protected in such a way as to
expose the hydroxyl group on which to add the phosphate group while
protecting all the remaining hydroxyls and other functional groups
on the nucleoside that may interfere with the phosphorylation step
or lead to regioisomers. In one aspect, the protecting groups
selected are resistant to strong bases, e.g., ethers, silyl ethers
and ketals. In one aspect, the protecting groups are optionally
substituted MOM ethers, MEM ethers, trialkylsilyl ethers and
symmetrical ketals. In another aspect, the protecting groups are
t-butyldimethylsilyl ether and isopropylidene. Further protection
entails masking of the amino group of the base moiety, if present,
so as to eliminate any acidic protons. In one aspect the selected
N-protecting groups are selected from the groups of dialkyl
formamidines, mono and dialkyl imines, mono and diaryl imines. In
one aspect, the N-protecting groups are selected from the groups of
dialkyl formamidines and mono-alkyl imine and mono aryl imine. In
one aspect the mono-alkyl imine is benzylimine and the mono-aryl
imine is phenylimine. In another aspect, the N-protecting group is
a symmetrical dialkyl formamidine selected from the group of
dimethyl formamidine and diethyl formamidine.
[0538] Generation of the alkoxide of the exposed hydroxyl group on
the suitably protected nucleoside is accomplished with a base in an
aprotic solvent that is not base sensitive such as THF, dialkyl and
cyclic formamides, ether, toluene and mixtures of those solvents.
In one aspect, the solvents are DMF, DMA, DEF,
N-methylpyrrolidinone, THF, and mixture of those solvents.
[0539] Many different bases have been used for the phosphorylation
of nucleosides and non-nucleoside compounds with cyclic and acyclic
phosphorylating agents. For example trialkylamines such as
triethylamine (Roodsari et al., J. Org. Chem. 64(21), 7727 (1999))
or N,N-diisopropylethylamine (Meek et al., J. Am. Chem. Soc.
110(7), 2317 (1988)); nitrogen containing heterocyclic amines such
as pyridine (Hoefler et al., Tetrahedron 56(11), 1485 (2000)),
N-methylimidazole (Vankayalapati et al., J. Chem. Soc. Perk T 1 14,
2187(2000)), 1,2,4-triazole (Takaku et al., Chem. Lett. (5), 699
(1986)) or imidazole (Dyatkina et al., Tetrahedron Lett. 35(13),
1961 (1994)); organometallic bases such as potassium t-butoxide
(Postel et al., J. Carbohyd. Chem. 19(2), 171 (2000)), butyllithium
(Torneiro et al., J. Org. Chem. 62(18), 6344 (1977)),
t-butylmagnesium chloride (Hayakawa et al., Tetrahedron Lett.
28(20), 2259 (1987)) or LDA (Aleksiuk et al., J. Chem. Soc. Chem.
Comm. (1), 11 (1993)); inorganic bases such as cesium fluoride
(Takaku et al., Nippon Kagaku Kaishi (10), 1968 (1985)), sodium
hydride (Hanaoka et al., Heterocycles 23(11), 2927 (1985)), sodium
iodide (Stromberg et al., J. Nucleos. Nucleot. 6(5), 815 (1987)),
iodine (Stromberg et al., J. Nucleos. Nucleot. 6(5), 815 (1987)) or
sodium hydroxide (Attanasi et al., Phosphorus Sulfur 35(1-2), 63
(1988)); metals such as copper (Bhatia et al., Tetrahedron Lett.
28(3), 271 (1987)). However, no reaction or racemization at the
phosphorus stereogenic center was observed when coupling of
phosphorylating reagent was attempted using the previously
described procedures. Especially, no reaction was observed with
bases previously used with substituted cyclic phosphorylating agent
to give the corresponding cyclic phosphate in high yield such as
sodium hydride (Thuong et al., Bull. Soc. Chim. Fr. 667 (1974)),
pyridine (Ayral-Kaloustian et al., Carbohydr. Res. 187(1991)),
butyl-lithium (Hulst et al., Tetrahedron Lett. 1339 (1993)), DBU
(Merckling et al., Tetrahedron Lett. 2217 (1996)), triethylamine
(Hadvary et al., Helv. Chim. Acta, 1986, 69(8), 1862),
N-methylimidazole (Li et al., Tetrahedron Lett. 6615 (2001)) or
sodium methoxide (Gorenstein et al., J. Am. Chem. Soc. 5077
(1980)). It was found that the use of Grignard reagents promoted
phosphorylation with minimal epimerization of the phosphorus
center. In one aspect, Grignard reagents are alkyl and aryl
Grignards. In another aspect, the Grignard reagents are t-butyl
magnesium halides and phenyl magnesium halides. In another aspect,
the Grignard reagents are t-butylmagnesium chloride and
phenylmagnesium chloride.
[0540] In another aspect magnesium alkoxides are used to generate
the magnesium 5'-alkoxide of the nucleoside. In one aspect
magnesium alkoxides are selected from the group of
Mg(O-t-Bu).sub.2, and Mg(O-iPr).sub.2.
[0541] The protected prodrugs generated as described above are then
subjected to a deprotection step to remove all the protecting
groups using one of the many methods known to those skilled in the
art (Greene T. W., Protective Groups in Organic Chemistry, John
Wiley & Sons, New York (1999)) and that are compatible with the
stability of the phosphate prodrug. In one aspect, deprotection
reagents include fluoride salts to remove silyl protecting groups,
mineral or organic acids to remove acid labile protecting groups
such as silyl and/or ketals and N-protecting groups, if present. In
another aspect, reagents are tetrabutylammonium fluoride (TBAF),
hydrochloric acid solutions and aqueous TFA solutions. Isolation
and purification of the final prodrugs, as well as all
intermediates, are accomplished by a combination of column
chromatography and/or crystallization.
[0542] The sequence provides methods to synthesize single isomers
of compounds of Formula I. Due to the presence of a stereogenic
center at the carbon where V is attached on the cyclic phosphate
reagent, this carbon atom can have two distinct orientations,
namely R or S. As such the trans-phosphate reagent prepared from a
racemic diol can exist as either the S-trans or R-trans
configuration and results in a S-cis and R-cis prodrug mixture. The
reaction of the C'-S-trans-phosphate reagent generates the
C'-S-cis-prodrug of the nucleoside while reaction with the
C'-R-trans-phosphate reagent generates the C'-R-cis-prodrug.
Synthesis of 6-, 2'-, and/or 3'-Substituted Prodrugs:
[0543] Synthesis of 6-, 2'- and/or 3-' substituted prodrugs of
Formula II or III can be accomplished starting from compounds of
Formula I. For example, selective 3'-acylation of nucleoside
monophosphate cyclic prodrugs of Formula I may be achieved by
several methods as described in the literature (Protective groups
in organic synthesis, Greene and Wuts, John Wiley, New York, 1991).
Additionally, selective 3'-acylation can be attained by various
esterification methods in the presence of tertiary hydroxy
functionality at the 2'-position without protection. Acylation may
also be accomplished efficiently by utilizing amine protected amino
acids as described earlier (WO 04/002422, Hanson et al., Bioorg.
Med. Chem. 2000, 8, 2681) and the amine protective groups are
removed under mild acidic conditions. 2',3-Cyclic carbonate
formation is another well-known transformation for ribofuranosyl
nucleosides. Compounds of formula I undergo carbonate formation
under neutral conditions to result in compounds of Formula II or
III (Pankiewicz, et al., J. Org. Chem., 1985, 50, 3319).
[0544] Prodrugs at 6-position may be prepared from the
corresponding halo derivatives of the nucleosides. The prodrug
substitution is made at the nucleoside stage (before 5'-prodrug
formation) from the corresponding chloro or hydroxy functionalities
in case of compounds of Formula II or III where R.sup.9 or R.sup.10
is substituted (e.g., N.sub.3, H, --COR). Synthesis of such
nucleoside precursors are attained as described earlier (WO
02/057287). Preparation of these purine analogs by azido
displacement (Aso et al., J. Chem Soc Perkin Trans II, 2000, 8
1637) or hydrogention (Freer et al., Tetrahedron, 2000, 56, 45) are
well known methods. Subsequently, these prodrug functionality
substituted nucleosides are transformed to corresponding
monophosphate cyclic prodrugs of Formula II or III.
Miscellaneous Methods:
[0545] Coupling of activated phosphates with alcohols is
accomplished in the presence of an organic base. For example,
chlorophosphates synthesized as described in the earlier section
react with an alcohol in the presence of a base such as pyridine or
N-methylimidazole. In some cases phosphorylation is enhanced by in
situ generation of iodophosphate from chlorophosphate (Stomberg, et
al., Nucleosides & Nucleotides., 1987, 5: 815).
Phosphorofluoridate intermediates have also been used in
phosphorylation reactions in the presence of a base such as CsF or
n-BuLi to generate cyclic prodrugs (Watanabe et al., Tetrahedron
Lett., 1988, 29, 5763). Phosphoramidate intermediates are known to
couple by transition metal catalysis (Nagamatsu, et al.,
Tetrahedron Lett., 1987, 28, 2375).
[0546] Reaction of the optically pure diastereomer of
phosphoramidate intermediate with the hydroxyl of nucleoside in the
presence of an acid produces the optically pure phosphate prodrug
by direct S.sub.N2(P) reaction (Nakayama, et al., J. Am. Chem.
Soc., 1990, 112, 6936). Alternatively, reaction of the optically
pure phosphate precursor with a fluoride source, preferably cesium
fluoride or TBAF, produces the more reactive phosphorofluoridate
which reacts with the hydroxyl of the nucleoside to give the
optically pure prodrug by overall retention of configuration at the
phosphorus atom (Ogilvie, et al., J. Am. Chem. Soc., 1977, 99,
1277).
[0547] Prodrugs of Formula I are synthesized by reaction of the
corresponding phosphodichloridate and an alcohol (Khamnei, et al.,
J. Med. Chem., 1996, 39: 4109). For example, the reaction of a
phosphodichloridate with substituted 1,3-diols in the presence of
base (such as pyridine and triethylamine) yields compounds of
Formula I.
[0548] Such reactive dichloridate intermediates can be prepared
from the corresponding acids and the chlorinating agents such as
thionyl chloride (Starrett, et al, J. Med. Chem., 1994, 1857),
oxalyl chloride (Stowell, et al., Tetrahedron Lett., 1990, 31:
3261), and phosphorus pentachloride (Quast, et al., Synthesis,
1974, 490).
[0549] Phosphorylation of an alcohol is also achieved under
Mitsunobu reaction conditions using the cyclic 1',3'-propanyl ester
of phosphoric acid in the presence of triphenylphosphine and
diethyl azodicarboxylate (Kimura et al., Bull. Chem. Soc. Jpn.,
1979, 52, 1191). The procedure can be extended to prepare
enantiomerically pure phosphates from the corresponding phosphoric
acids. Phosphate prodrugs are also prepared from the free acid by
Mitsunobu reactions (Mitsunobu, Synthesis, 1981, 1; Campbell, J.
Org. Chem., 1992, 52: 6331), and other acid coupling reagents
including, but not limited to, carbodiimides (Alexander, et al.,
Collect. Czech. Chem. Commun., 1994, 59: 1853; Casara, et al.,
Bioorg. Med. Chem. Lett., 1992, 2: 145; Ohashi, et al., Tetrahedron
Lett., 1988, 29: 1189), and
benzotriazolyloxytris-(dimethylamino)phosphonium salts (Campagne,
et al., Tetrahedron Lett., 1993, 34: 6743). Cyclic-1,3-propanyl
prodrugs of phosphates are also synthesized from NMP and
substituted propane-1,3-diols using a coupling reagent such as
1,3-dicyclohexylcarbodiimide (DCC) in presence of a base (e.g.,
pyridine). Other carbodiimide based coupling agents such as
1,3-diisopropylcarbodiimide and the water soluble reagent,
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI)
can also be utilized for the synthesis of cyclic prodrugs.
[0550] Phosphate prodrugs can be prepared by an alkylation reaction
between the phosphate corresponding tetrabutylammonium salts and
substituted-1,3-diiodopropanes made from 1,3-diols (Farquhar, et
al., Tetrahedron Lett., 1995 36, 655). Furthermore, phosphate
prodrugs can be made by conversion of nucleoside to the
dichloridate intermediate with phosphoryl chloride in presence of
triethylphosphite and quenching with substituted-1,3-propanediols
(Farquhar et al., J. Org. Chem., 1983, 26, 1153).
[0551] Phosphorylation can also be achieved by making the mixed
anhydride of the cyclic diester of phosphoric acid and a sulfonyl
chloride, preferably 8-quinolinesulfonyl chloride, and reacting the
hydroxyl of the nucleoside in the presence of a base, preferably
N-methylimidazole (Takaku, et al., J. Org. Chem., 1982, 47, 4937).
In addition, starting from an enantiomerically pure cyclic diester
of a phosphoric acid, obtained by resolution (Wynberg, et al., J.
Org. Chem., 1985, 50, 4508), one can obtain enantiomerically pure
phosphates.
EXAMPLES
[0552] The compounds used in this invention and their preparation
can be understood further by the Examples, which illustrate some of
the processes by which these compounds are prepared. These Examples
should not however be construed as specifically limiting the
invention, and variations of the compounds, now known or later
developed, are considered to fall within the scope of the present
invention as hereinafter claimed.
[0553] Compounds of Formula I are prepared as outlined below. The
TLC conditions given are utilizing plates of Analtech UNIPLATE,
silica gel GHLF, scored 10.times.20 cm, 250 micron.
Synthesis of Racemic 1-(Aryl)Propane-1,3-Diols
Example 1
Preparation of 1-(2'-Furanyl)-Propane-1,3-Diol Via Grignard
Addition and Hydroboration
[0554] To a solution of 2-furaldehyde (3 g, 31.2 mmol) in THF (60
mL) was added 1 M vinyl magnesium bromide in THF (34 mL) at
0.degree. C. After stirring for an hour, a solution of 1 M BH.sub.3
THF complex in THF was added. The reaction was quenched with 3N
NaOH (20 mL) and 30% hydrogen peroxide (10 mL) at 0.degree. C. The
organic fraction was separated and concentrated. The crude product
was chromatographed by eluting with 5% methanol-dichloromethane to
give 1-(2'-furyl)propane-1,3-diol (1 g).
Example 2
Preparation of 1-(2'-Pyridyl)-Propane-1,3-Diol Via Benzylic
Oxidation
Step A: (J. Org. Chem. 22:589 (1957))
[0555] To a solution of 3-(2'-pyridyl)propan-1-ol (10 g, 72.9 mmol)
in acetic acid (75 mL) was added 30% hydrogen peroxide slowly. The
reaction mixture was heated to 80.degree. C. for 16 h. The reaction
was concentrated under vacuum and the residue was dissolved in
acetic anhydride (100 mL) and heated at 110.degree. C. overnight.
Acetic anhydride was evaporated upon completion of the reaction.
Chromatography of the mixture by eluting with methanol-methylene
chloride (1:9) resulted in 10.5 g of pure diacetate.
Step B:
[0556] To a solution of diacetate (5 g, 21.1 mmol) in
methanol-water (3:1, 40 mL) was added potassium carbonate (14.6 g,
105.5 mmol). After stirring for 3 h at room temperature, the
reaction mixture was concentrated. The residue was chromatographed
by eluting with methanol-methylene chloride (1:9) to give 2.2 g of
crystalline diol.
Example 3
Preparation of 1-(Aryl)-Propane-1,3-Diol from Propane-1,3-Diol Via
Grignard Addition
Step A: (J. Org. Chem. 53:911 (1988))
[0557] To a solution of oxalyl chloride (5.7 mL, 97 mmol) in
dichloromethane (200 mL) at -78.degree. C. was added dimethyl
sulfoxide (9.2 mL, 130 mmol). The reaction mixture was stirred at
-78.degree. C. for 20 min before addition of
3-(benzyloxy)propan-1-ol (11 g, 65 mmol) in dichloromethane (25
mL). After an hour at -78.degree. C., reaction was quenched with
triethylamine (19 mL, 260 mmol) and warmed to room temperature.
Work-up and column chromatography by elution with dichloromethane
resulted in 8 g of 3-(benzyloxy)propan-1-al.
Step B:
[0558] To a solution of 3-(benzyloxy)propan-1-al (1 g, 6.1 mmol) in
THF at 0.degree. C. was added a 1 M solution of
4-fluorophenylmagnesium bromide in THF (6.7 mL, 6.7 mmol). The
reaction was warmed to room temperature and stirred for 1 h.
Work-up and column chromatography by elution with dichloromethane
resulted in 0.7 g of alcohol.
Step C:
[0559] To a solution of benzyl ether (500 mg) in ethyl acetate (10
mL) was added 10% Pd(OH).sub.2C (100 mg). The reaction was stirred
under hydrogen gas for 16 h. The reaction mixture was filtered
through Celite and concentrated. Chromatography of the residue by
elution with ethyl acetate-dichloromethane (1:1) resulted in 340 mg
of product.
Example 4
General Procedure for Preparation of 1-Aryl
Substituted-Propane-1,3-Diol from Aryl Aldehyde
Step A: (J. Org. Chem. 55:4744 (1990))
[0560] To a -78.degree. C. solution of diisopropylamine (2 mmol) in
THF (0.7 mL/mmol diisopropylamine) was slowly added n-butyllithium
(2 mmol, 2.5 M solution in hexanes). The reaction was then stirred
for 15 min at -78.degree. C. before a solution of ethyl acetate (2
mmol) in THF (0.14 mL/mmol ethyl acetate) was slowly introduced.
After stirring an additional 30 min at -78.degree. C., a THF
solution containing the aryl aldehyde (1.0 mmol in 0.28 mL THF) was
added. The reaction was then stirred at -78.degree. C. for 30 min,
warmed to room temperature and stirred an additional 2 h. After
aqueous work up (0.5 M HCl), the organic layer was concentrated to
a crude oil (beta-hydroxyester).
Step B:
[0561] The crude hydroxyester was dissolved in ether (2.8 mL/mmol),
cooled to ice bath temperature, and lithium aluminum hydride (3
mmol) was added batch wise. The reaction was stirred allowing the
cooling bath to melt and the reaction to reach room temperature.
After stirring overnight at room temperature, the reaction was
cooled back to ice bath temperature and quenched with ethyl
acetate. Aqueous work up (0.5 M HCl) afforded the crude diol, which
was purified either by chromatography or distillation.
Example 4a
Synthesis of 1-(3-methoxycarbonylphenyl)-1,3-propanediol
[0562] 1-(3-bromophenyl)-1,3-propane diol was prepared as Example 4
and further derivatized as follows:
[0563] A pressure vessel was charged with
1-(3-bromophenyl)-1,3-propanediol (2 g, 8.6 mmol), methanol (30
mL), triethylamine (5 mL) and bis(triphenylphosphine)palladium
dichloride (0.36 g, 05 mmol). The sealed vessel was pressurize with
carbon monoxide at 55 psi and heated at 85.degree. C. for 24 h. The
cooled vessel was opened and the reaction mixture was filtered
through Celite and rinsed with methanol. The combined filtrates
were concentrated under reduced pressure and the residue was
purified by column chromatography (silica gel, hexanes/ethyl
acetate 1/1) to afford the title compound (1.2 g)
[0564] TLC: hexanes/ethyl acetate 2/8; Rf=0.5
[0565] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): 5.05-4.95
(m, 1H), 3.9 (s, 3H), 2-1.8 (m, 2H).
Example 4b
Synthesis of 1-(4-methoxycarbonylphenyl)-1,3-propanediol
[0566] 1-(4-bromophenyl)-1,3-propane diol was prepared as Example 4
and further derivatized as Example 4a.
[0567] TLC: hexanes/ethyl acetate 3/7; Rf=0.35
[0568] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): 5.1-5 (m,
1H), 3.91 (s, 3H), 2.05-1.9 (m, 2H).
Synthesis of Enantioenriched 1-(Aryl)-Propane-1,3-Diols:
Example 5
General Procedure for Resolution of Racemic 1,3-diols
[0569] Racemic diols synthesized as in Examples 1-4 may be resolved
to yield both enantiomers as described in the following
procedure.
Step A:
[0570] To a solution of diol (1.0 mmol) in THF (1.0 ml) was added
hexamethyldisilazane (2.1 mmol) followed by a catalytic amount of
trimethylsilyltriflate (2-3 drops). After stirring at room
temperature for 1 h, the reaction was diluted with hexane (4 mL)
and subjected to Work up with ice-cold water. The resulting
disilylether was either purified by chromatography or, if
sufficiently pure, used crude in the next reaction.
Step B:
[0571] To a solution of disilylether (1.0 mmol) and (-)-menthone
(1.1 mmol) in dichloromethane (2.0 ml) at -40.degree. C., was
slowly added trimethylsilyltriflate (0.11 mmol). The reaction was
then kept at -50.degree. to -60.degree. C. for 48 h, at which time
pyridine was added to quench the reaction. After warming to room
temperature, the crude mixture was diluted with hexane (4.0 ml) and
subjected to aqueous work up. The two ketals were separated by
chromatography.
Step C:
[0572] The separated ketals were hydrolyzed by adding a catalytic
amount of concentrated hydrochloric acid to a methanol (4.0 mmol)
solution of each. After stirring overnight at room temperature, the
methanol was removed under vacuum and the residue was subjected to
aqueous work up. The resolved diols were further purified by either
chromatography or distillation.
Example 6
Synthesis of Enantioenriched
1-(3'-chlorophenyl)-1,3-dihydroxypropane Via Sharpless Asymmetric
Epoxidation
Step A:
[0573] To a dispersion of m-chloro-cinnamic acid (25 g, 137 mmol)
in ethanol (275 mL) was added conc. sulfuric acid (8 mL) at room
temperature. The reaction was refluxed overnight and concentrated.
Ice-cold water was added to the crude and precipitated white solid
was filtered and washed with cold water. The precipitate was dried
under vacuum overnight to give 25 g of ester. (Rf=0.5 in
dichloromethane on silica)
Step B:
[0574] To a solution of ethyl-m-chlorocinnamate (23 g, 109.5 mmol)
in dichloromethane at -78.degree. C. was added 1 M DIBAL-H in
dichloromethane (229 mL, 229 mmol) dropwise over 1 h. The reaction
was stirred at -78.degree. C. for an additional 3 h. Ethylacetate
was added to quench excess DIBAL-H and saturated aq. potassium
sodium tartrate was added and the reaction was stirred at room
temperature for 3 h. The organic layer was separated and salts were
washed with ethyl acetate. The combined organic extracts were
concentrated and distilled at 120.degree. C./0.1 mm to give 14 g of
pure allylic alcohol. (Rf=0.38 in 1:1 ethylacetate:hexane on
silica)
Step C:
[0575] To a solution of m-chlorocinnamyl alcohol (5 g, 29.76 mmol)
in dichloromethane (220 mL) was added activated 4 .ANG. molecular
sieves powder (2.5 g) and the mixture was cooled to -20.degree. C.
(+)-Diethyl tartrate (0.61 mL, 3.57 mmol) was added at -20.degree.
C. and stirred for 15 min before adding titanium tetraisopropoxide
(0.87 g, 2.97 mmol). The reaction was stirred for additional 30 min
and 5-6 M solution of t-butylhydroperoxide in heptane (10 mL, 60
mmol) was added dropwise while maintaining the internal temperature
at -20 to -25.degree. C. The mixture was stirred for an additional
3 h at -20.degree. C. and a 10% sodium hydroxide in saturated aq.
sodium chloride (7.5 mL) followed by ether (25 mL) were added. The
reaction was warmed to 10.degree. C. and stirred for 15 min before
adding anhydrous magnesium sulfate (10 g) and Celite (1.5 g). The
mixture was further stirred for additional 15 min, filtered and
concentrated at 25.degree. C. to give crude epoxy alcohol. (R=0.40
in 1:1 ethylacetate:hexane on silica)
Step D:
[0576] To a solution of crude m-chloroepoxycinnamyl alcohol
obtained from earlier reaction in dimethoxyethane (300 mL) was
added a 65% Red-Al solution in toluene (18.63 mL, 60 mmol) dropwise
under nitrogen at 0.degree. C. After stirring at room temperature
for 3 h, the solution was diluted with ethyl acetate (400 mL) and
quenched with aq. saturated sodium sulfate solution (50 mL). After
stirring at room temperature for 30 min, the resulting white
precipitate formed was filtered and washed with ethylacetate. The
filtrate was dried and concentrated. The crude product was
distilled at 125-130.degree. C./0.1 mm to give 3.75 g of
enantioenriched (R)-1-(3'-chlorophenyl)-1,3-dihydroxypropane.
(Rf=0.40 in 1:1 ethylacetate:dichloromethane)
[0577] Enantiomeric excesses were defined as diacetates (prepared
by treatment of diols with acetic anhydride, triethylamine,
cat.DMAP in dichloromethane) by HPLC ((S,S) Whelko-0, 250
cm.times.4.0 mm ID purchased from Regis).
[0578] (R)-1-(3'-chlorophenyl)-1,3-dihydroxypropane: 91% ee
[0579] (+)Diisopropyltartrate provided >96% ee in
(R)-1-(3'-chlorophenyl)-1,3-dihydroxypropane.
[0580] (S)-1-(3'-chlorophenyl)-1,3-dihydroxypropane was also
prepared under identical conditions via asymmetric epoxidation and
reduction protocol utilizing (-)-tartrate in similar yields.
(S)-3-(3'-chlorophenyl)-1,3-dihydroxypropane was obtained with 79%
ee.
Example 7
Synthesis of Enantioenriched
1-(3'-chlorophenyl)-1,3-hihydroxypropane Via Hydrogen Transfer
Reaction
Step A: Preparation of Methyl
3-(3'-chlorophenyl)-3-oxo-propanoate:
[0581] A 22 L, 3-neck round bottom flask was equipped with a
mechanical stirrer, thermowell/thermometer and nitrogen inlet
(bubbler in-line). The flask was flushed with nitrogen and charged
sequentially with THF (6 L), potassium t-butoxide (1451 g), and THF
(0.5 L). The resulting mixture was stirred at ambient temperature
for 15 min. and a 20.degree. C. water bath was applied. A 3 L round
bottom flask was charged with 3'-chloroacetophenone (1000 g) and
diethylcarbonate (1165 g), and the resulting yellow solution was
added slowly to the stirred potassium t-butoxide solution,
maintaining the temperature between 16 and 31.degree. C. After the
addition was complete (1 h, 10 min.), the cooling bath was removed
and the solution was stirred for 1 h, 30 min. TLC indicated that
the reaction was complete. A 5 gallon stationary separatory funnel
was charged with ice water (4 L) and concentrated hydrochloric acid
(1.3 L of 12 M solution). The dark red reaction solution was
quenched into the aqueous acid and the mixture was stirred for 15
min. The layers were separated and the aqueous phase (lower) was
extracted again with toluene (4 L). The combined organic extracts
were washed with saturated brine (2.times.3 L, 10 min. stirring
time each), dried (MgSO.sub.4), filtered and concentrated under
reduced pressure to provide 1480 g of a brown oil. The oil was
placed under high vacuum (10 torr) overnight to give 1427 g. The
material was vacuum distilled (short path column, fraction cutter
receiver) and the fraction at 108-128.degree. C./1-0.5 torr was
collected to provide 1273.9 g of a yellow oil. (Rf=0.36 in 20%
ethyl acetate/hexanes). Step B: Preparation of Methyl
(S)-3-(3'-chlorophenyl)-3-hydroxypropionate: ##STR63##
[0582] A 12 L, 3-neck round bottom flask was equipped with a
mechanical stirrer, thermometer, addition funnel (500 mL) and
nitrogen inlet (bubbler in-line). The flask was flushed with
nitrogen and charged with formic acid (292 mL, 350 g).
Triethylamine (422 mL, 306 g) was charged to the addition funnel,
then added slowly with stirring, maintaining the temperature less
than 45.degree. C. After the addition was complete (1 h, 30 min),
the solution was stirred with the ice bath applied for 20 min.,
then at ambient temperature for an additional 1 h. The flask was
charged sequentially with methyl
3-(3-chlorophenyl)-3-oxo-propanoate (1260 g), DMF (2.77 L including
rinsing volume) and (S,S)-Ts-DPEN-Ru--Cl-(p-cymene) (3.77 g). The
flask was equipped with a heating mantle and the addition funnel
was replaced with a condenser (5 C circulating coolant for
condenser). The stirred reaction solution was slowly heated to
60.degree. C. (90 min. to attain 60.degree. C.) and the contents
were maintained at 60.degree. C. for 4.25 h. HPLC indicated 3%
starting material remained. The solution was stirred at 60.degree.
C. for an additional 8 h, then gradually cooled to ambient
temperature overnight. HPLC indicated 0.5% starting material. A 5
gallon stationary separatory funnel was charged with water (10 L)
and MTBE (1 L). The reaction solution was poured into the aqueous
mixture and the reaction flask was rinsed into the separatory
funnel with an additional 1 L of MTBE. The contents were stirred
for several minutes and the layers were separated. The aqueous
phase was extracted with additional MTBE (2.times.1 L), and the
combined organic extracts were washed with brine (1 L), and
concentrated under reduced pressure to provide 1334 g of a red oil.
The oil was used without further purification for the next
step.
[0583] The crude hydroxyester (10 mg, 0.046 mmol) was dissolved in
dichloromethane (1 mL). Acetic anhydride (22 .mu.L, 0.23 mmol) and
4-(dimethylamino)pyridine (22 mg, 0.18 mmol) were added and the
solution was stirred at ambient temperature for 15 min. The
solution was diluted with dichloromethane (10 mL) and washed with 1
M hydrochloric acid (3.times.3 mL). The organic phase was dried
(MgSO.sub.4), filtered and concentrated under reduced pressure. The
residual oil was dissolved in methanol and analyzed by chiral HPLC
(Zorbax Rx-C18, 250.times.4.6 mm; mobile phase: 65/35 (v/v)
water/acetonitrile, isocratic; flow rate=1.5 mL/min; inj. volume=15
.mu.L; UV detection at 220 nm. Retention times: Product=9.3 min,
starting material=17.2 min). The hydroxyester was derivatized to
the acetate for analysis by chiral HPLC and shown to give 91% ee.
(HPLC conditions: Column: Pirkle covalent (S,S) Whelk-O 10/100 krom
FEC, 250.times.4.6 mm; mobile phase: 70/30 (v/v) methanol/water,
isocratic; flow rate: 1.5 mL/min; inj. volume=10 .mu.L; UV
detection at 220 nm. Retention times: S-hydroxyester (acetate)=9.6
min, R-hydroxyester (acetate)=7.3 min.)
Step C: Preparation of (S)-3-(3'-chlorophenyl)-3-hydroxypropanoic
Acid:
[0584] To the crude hydroxyester in a 10 L rotary evaporator flask
was added sodium hydroxide solution (2.5 L of 2 M solution). The
resulting solution was stirred on the rotary evaporator at ambient
pressure and temperature for 2 h. HPLC indicated 5% starting
material still remained (HPLC conditions: Column: Zorbax Rx-C18,
250.times.4.6 mm; mobile phase: 65/35 (v/v) water/acetonitrile,
isocratic; flow rate=1.5 mL/min; inj. volume=15 .mu.L; UV detection
at 220 nm. Retention times: Product=3.8 min, starting material=18.9
min.). The pH of the solution was 11 (wide range pH paper).
Additional 2 M NaOH solution was added to adjust the pH to 14
(approx. 100 mL), and the solution was stirred for an additional 30
min. HPLC indicated the reaction was complete. The solution was
transferred to a 5 gallon stationary separatory funnel and
extracted with MTBE (2 L). The layers were separated and the
organic extract was discarded. The aqueous phase was transferred
back to the separatory funnel and acidified with 12 M HCl solution
(600 mL). The mixture was extracted with MTBE (1.times.2 L,
2.times.1 L). The combined acidic organic extracts were dried
(MgSO.sub.4), filtered and concentrated under reduced pressure to
give 1262 g of a brown, oily semi-solid. The residue was slurried
with ethyl acetate (1 L) and transferred to a 12 L, 3-neck round
bottom flask equipped with a mechanical stirrer, heating mantle,
condenser and thermometer. The stirred mixture was heated to
dissolve all solids (28.degree. C.) and the dark solution was
cooled to 10.degree. C. (a precipitate formed at 11.degree. C.).
The mixture was slowly diluted with hexanes (4 L over 1 h) and the
resulting mixture was stirred at <10.degree. C. for 2 h. The
mixture was filtered and the collected solid was washed with cold
4/1 hexanes/ethyl acetate (1 L), and dried to constant weight (-30
in. Hg, 50.degree. C., 4 h). Recovery=837 g of a beige solid.
mp=94.5-95.5.degree. C.
[0585] A 50 mg sample of hydroxyacid was reduced to the diol with
borane-THF (see Step D). The resulting crude diol was diacetylated
(as described in Step B)) and analyzed by chiral HPLC. Retention
times: S-diol (diacetate)=12.4 min, R-diol (diacetate)=8.8 min.)
ee=98%
[0586] A second crop of hydroxyacid was isolated. The filtrate from
above was concentrated under reduced pressure to give 260 g of a
brown sludge. The material was dissolved in ethyl acetate (250 mL)
and the stirred dark solution was slowly diluted with hexanes (1000
mL) and the resulting mixture was stirred at ambient temperature
overnight. The mixture was filtered and the collected solid was
washed with 5/1 hexanes/ethyl acetate (200 mL), and dried to
constant weight (-30 in. Hg, 50.degree. C., 16 h). Recovery=134 g
of a beige solid. ee=97% Step D: Preparation of
(S)-(-)-1-(3-chlorophenyl)-1,3-propanediol: ##STR64##
[0587] A 22 L, 3-neck round bottom flask was equipped with a
mechanical stirrer, thermowell/thermometer and nitrogen inlet
(outlet to bubbler). The flask was charged with 2 M borane-THF
(3697 g, 4.2 L) and the stirred solution was cooled to 5.degree. C.
A solution of (S)-3-(3-chlorophenyl)-3-hydroxypropanoic acid (830
g) in THF (1245 mL) was prepared with stirring (slightly
endothermic). The reaction flask was equipped with an addition
funnel (1 L) and the hydroxyacid solution was slowly added to the
stirred borane solution, maintaining the temperature
.ltoreq.16.degree. C. After the addition was complete (3 h), the
mixture was stirred at ice bath temperature for 1.5 h. The reaction
was quenched by careful addition of water (2.5 L). After the
addition was complete (30 min), 3 M NaOH solution (3.3 L) was added
(temperature increased to 35.degree. C.) and the resulting mixture
was stirred for an additional 20 min. (temperature=30.degree. C.).
The reaction mixture was transferred to a 5 gallon stationary
separatory funnel and the layers were separated. The aqueous phase
was extracted with MTBE (2.5 L) and the combined organic extracts
(THF and MTBE) were washed with 20 wt % NaCl solution (2 L) and
stirred with MgSO.sub.4 (830 g) for 30 min. The mixture was
filtered through Celite and concentrated under reduced pressure to
provide 735 g of a thick, brown oil.
[0588] The oil was purified by vacuum distillation and the fraction
at 135-140.degree. C./0.2 mm Hg was collected to provide 712.2 g of
a colorless oil.
[0589] The diol was diacetylated and analyzed by chiral HPLC
(e.e.=98%) (see Step B). Retention times: S-diol (diacetate)=12.4
min, R-diol (diacetate)=8.9 min. [.alpha.].sub.D=-51.374 (5 mg/mL
in CHCl.sub.3)
Example 8
Synthesis of Enantioenriched 1-(4'-pyridyl)-1,3-Dihydroxypropane
Via Hydrogen Transfer Reaction
Step A: Synthesis of Methyl 3-oxo-3-(pyridin-4-yl)-propanoate
[0590] A 50 L, 3-neck flask was equipped with an overhead stirrer,
heating mantle, and nitrogen inlet. The flask was charged with THF
(8 L), potassium t-butoxide (5 kg, 44.6 mol), and THF (18 L).
4-Acetylpyridine (2.5 kg, 20.6 mol) was added, followed by
dimethylcarbonate (3.75 L, 44.5 mol). The reaction mixture was
stirred without heating for 2.5 h then with heating to
57-60.degree. C. for 3 h. The heat was turned off and the mixture
cooled slowly overnight (15 h). The mixture was filtered through a
45 cm Buchner funnel. The solid was returned to the 50 L flask and
diluted with aqueous acetic acid (3 L acetic acid in 15 L of
water). The mixture was extracted with MTBE (1.times.16 L,
1.times.12 L). The combined organic layers were washed with aqueous
Na.sub.2CO.sub.3 (1750 g in 12.5 L water), saturated aqueous
NaHCO.sub.3 (8 L), and brine (8 L) then dried over MgSO.sub.4 (500
g) overnight (15 h). The solution was filtered and the solvent
removed by rotary evaporation to a mass of 6.4 kg. The resulting
suspension was cooled in an ice bath with stirring for 2 h. The
solid was collected by filtration, washed with MTBE (500 mL), and
dried in a vacuum oven at 20.degree. C. for 15 h, giving 2425 g of
the keto ester as a pale yellow solid.
[0591] The MTBE mother liquor was concentrated to approximately 1
L. The resulting suspension was cooled in an ice bath for 1 h. The
solid was collected by filtration, washed with MTBE (2.times.150
mL), and dried in a vacuum oven to give 240 g of a second crop.
[0592] TLC. Merck silica gel plates, 1:2 THF/hexane, UV lamp, Rf of
SM=0.25, Rf of product=0.3.
[0593] Melting Point: 74-76.degree. C. Step B: Synthesis of
S-methyl-3-hydroxy-3-(pyridin-4-yl)-propanoate ##STR65##
[0594] A 22 L, 3-neck round bottom flask was equipped with an
overhead stirrer, thermowell/thermometer, addition funnel (1 L),
and cooling vessel (empty). The flask was flushed with nitrogen,
charged with formic acid (877 g) and cooled with an ice bath.
Triethylamine (755 g) was charged to the addition funnel and added
slowly over 50 min. to the stirred formic acid. After the addition
was complete, the cooling bath was removed and the reaction
solution was diluted with DMF (5.0 L). The ketoester (2648 g) was
added in one portion, followed by an additional 0.5 L of DMF. The
flask was equipped with a heating mantle and the stirred mixture
was heated gradually to 16.degree. C. to dissolve all solids. The
catalyst (S,S)-Ts-DPEN-Ru--Cl-(p-cymene) (18.8 g) was added in one
portion and the stirred mixture was heated to 55.degree. C. over 1
h. The resulting dark solution was stirred at 55.degree. C. for 16
h. TLC indicated the reaction was complete. The solvent was
evaporated under reduced pressure (Buchi R152 rotary evaporator
under high vacuum, bath temp=60.degree. C.) to give 3574 g of a
brown oil. The oil was dissolved in dichloromethane (10 L) and
transferred to a 5 gal. stationary separatory funnel. The dark
solution was washed with saturated sodium bicarbonate solution (3.0
L) and the aqueous phase was back extracted with dichloromethane
(3.0 L). The combined dichloromethane extracts were dried over
MgSO.sub.4 (300 g), filtered, and concentrated under reduced
pressure to provide 3362 g of a brown oil.
[0595] Column: Chiralpak AD, 0.46.times.25 cm; mobile phase=10:90,
ethanol:hexane, isocratic; flow rate=1.5 mL/min; injection
volume=10 .mu.L UV detection at 254 nm.
[0596] Retention times: R-hydroxy ester=19.9 min. [0597] S-hydroxy
ester=21.7 min.
[0598] Retention times: R-diol=14.2 min. [0599] S-diol=15.5 min
[0600] Hydroxy Ester:
[0601] .sup.1H NMR (CDCl.sub.3): .delta. 2.73 (d, 2H, J=1.5 Hz),
3.73 (s, 3H), 4.35 (s, 1H), 5.11-5.19 (m, 1H), 7.31 (d, 2H, J=6.6
Hz), 8.53 (d, 2H, J=6.0 Hz)
[0602] Merck silica gel 60 plates, 2.5.times.7.5 cm, 250 micron; UV
lamp: 5% MeOH in CH.sub.2Cl.sub.2; Rf of S.M.=0.44, Rf of
product=0.15.
[0603] e.e.=87% S isomer of hydroxy ester. Step C: Synthesis of
S-(-)-1-(Pd-4-yl)-1,3-propanediol ##STR66##
[0604] A 22 L, 4-neck round bottom flask was equipped with an
overhead stirrer, thermowell/thermometer, addition funnel (2 L),
condenser and cooling vessel (empty). The flask was flushed with
nitrogen and charged sequentially with sodium borohydride (467 g,
12.3 mol), 1-butanol (9.0 L), and water (148 mL, 8.23 mol) The
crude hydroxyester was dissolved in 1-butanol (1.0 L) and the
solution was charged to the addition funnel. The solution was added
over 3.25 h, using cooling as necessary to keep the temperature
below 62.degree. C. After addition was complete, the mixture was
stirred for 0.5 h then the flask was equipped with a heating mantle
and the stirred mixture was heated to 90.degree. C. over 0.75 h.
The mixture was stirred at 90-93.degree. C. for 2.25 h, then cooled
over 1.5 h to 28.degree. C. The reaction mixture was quenched with
aqueous potassium carbonate solution (10 wt/vol %, 6 L) and the
mixture was stirred for 10 min. The layers were separated and the
butanol phase was washed with aqueous potassium carbonate solution
(10 wt/vol %, 2 L) and sodium chloride solution (15 wt/vol %, 2 L).
The solvent was removed under reduced pressure (Buchi R152 rotary
evaporator, high vacuum, bath temperature=60.degree. C.) until a
concentrated solution resulted and 10.5 L of distillate had been
collected. Acetonitrile (3 L) was fed into the evaporator flask and
the solvent was evaporated under reduced pressure. Acetonitrile (9
L) was again fed into the evaporator flask and the slurry was
stirred (rotation on the rotary evaporator) at .about.60.degree. C.
(bath temperature=70.degree. C., atmospheric pressure) for 15 min.
The hot slurry was filtered through Celite 521 (250 g as a slurry
in 1 L of acetonitrile was prepacked on a 24 cm Buchner funnel).
The filtrate was partially concentrated under reduced pressure (5 L
of distillate were collected) and the resulting slurry was heated
at atmospheric pressure on the rotary evaporator to dissolve all
solids (bath temp=65.degree. C.). The heat source was turned off
and the resulting solution was stirred on the rotary evaporator for
10 h, with gradual cooling to ambient temperature. The resulting
mixture was filtered and the collected solid was washed with
acetonitrile (2.times.200 mL) and dried to constant weight (-30 in.
Hg, 55.degree. C., 4 h), giving S-(-)-1-(4-pyridyl)-1,3-propanediol
as a yellow solid weighing 496 g.
[0605] Melting point=98-100.degree. C.
[0606] HPLC conditions:
[0607] Column: Chiralpak AD, 0.46.times.25 cm; mobile phase=10:90,
ethanol:hexane, isocratic; flow rate=1.5 mL/min; injection
volume=10 .mu.L UV detection at 254 nm. [0608] Retention times:
R-diol=14.2 min. [0609] S-diol=15.5 min.
[0610] Merck silica gel 60 plates, 2.5.times.7.5 cm, 250 micron; UV
lamp; 15% MeOH in CH.sub.2Cl.sub.2; Rf of starting material=0.38,
Rf of product=0.17, Rf of boron complex=0.26.
Example 9
Synthesis of (S)-3-(3'-chlorophenyl)-1,3-dihydroxypropane Via
(-)-.beta.-chlorodiisopinocampheylborane (DIPCl) Reduction
Step A: Preparation of 3-(3-chlorophenyl)-3-oxo-propanoic Acid:
[0611] A 12 L, 3-neck round bottom flask was equipped with a
mechanical stirrer and addition funnel (2 L). The flask was flushed
with nitrogen and charged with diisopropylamine (636 mL) and THF
(1.80 L). A thermocouple probe was immersed in the reaction
solution and the stirred contents were cooled to -20.degree. C.
n-Butyllithium (1.81 L of a 2.5 M solution in hexanes) was charged
to the addition funnel and added slowly with stirring, maintaining
the temperature between -20 and -28.degree. C. After the addition
was complete (30 min), the addition funnel was rinsed with hexanes
(30 mL) and the stirred solution was cooled to -62.degree. C.
Trimethylsilyl acetate (300 g) was added slowly with stirring,
maintaining the temperature <-60.degree. C. After the addition
was complete (30 min), the solution was stirred at -60.degree. C.
for 15 min. 3-Chlorobenzoyl chloride (295 mL) was added slowly with
stirring, maintaining the temperature <-60.degree. C. After the
addition was complete (65 min), the cooling bath was removed and
the reaction solution was stirred for 1.25 h, with gradual warming
to 0.degree. C. The reaction flask was cooled with an ice bath,
then water (1.8 L) was added to the stirred solution. The reaction
mixture was stirred for 10 min., then diluted with t-butyl methyl
ether (1.0 L). The lower aqueous phase was separated and
transferred to a 12 L, 3-neck round bottom flask equipped with a
mechanical stirrer. t-Butyl methyl ether was added (1.8 L) and the
stirred mixture was cooled to <10.degree. C. (ice bath).
Concentrated HCl solution (300 mL of 12 M solution) was added and
the mixture was vigorously stirred. The layers were separated and
aqueous phase was further acidified with con. HCl (30 mL) and
extracted again with t-butyl methyl ether (1.0 L). The combined
MTBE extracts were washed with brine (1 L), dried (MgSO4, 70 g),
filtered and concentrated under reduced pressure to give 827 g of a
yellow solid. The crude solid was slurried in hexanes (2.2 L) and
transferred to a 5 L, 3-neck round bottom flask equipped with a
mechanical stirrer. The mixture was stirred at <10.degree. C.
(ice bath) for 1 h, then filtered, washed with hexanes (4.times.100
mL) and dried to constant weight (-30 in. Hg, ambient temperature,
14 h). Recovery=309 g of a pale yellow powder. Step B: Preparation
of (S)-3-(3-chlorophenyl)-3-hydroxypropanoic Acid: ##STR67##
[0612] A 12 L, 3-neck round bottom flask was equipped with a
mechanical stirrer and addition funnel (1 L). The flask was flushed
with nitrogen and charged with 3-(3-chlorophenyl)-3-oxo-propanoic
acid (275.5 g) and dichloromethane (2.2 L). A thermocouple probe
was immersed in the reaction slurry and the stirred contents were
cooled to -20.degree. C. Triethylamine (211 mL) was added over 5
min. to the stirred slurry and all solids dissolved. A
dichloromethane solution of
(-)-.beta.-chlorodiisopinocampheylborane (1.60 M, 1.04 L) was
charged to the addition funnel, then added slowly with stirring,
maintaining the temperature between -20 and -25.degree. C. After
the addition was complete (35 min), the solution was warmed to ice
bath temperature (2-3.degree. C.) and stirred for 4 h. Water (1.2
L) was added to the cloudy orange reaction mixture, followed by 3 M
NaOH solution (1.44 L). The mixture was vigorously stirred for 5
min, then transferred to a separatory funnel. The layers were
separated and the basic aqueous phase was washed with ethyl acetate
(1.0 L). The aqueous phase was acidified with conc. HCl (300 mL)
and extracted with ethyl acetate (2.times.1.3 L). The two acidic
ethyl acetate extracts were combined, washed with brine (600 mL),
dried (MgSO.sub.4, 130 g), filtered and concentrated under reduced
pressure to provide 328 g of a yellow oil (the oil crystallized on
standing). The solid was slurried in ethyl acetate (180 mL) and
transferred to a 2 L, 3-neck round bottom flask, equipped with a
mechanical stirrer. The stirred mixture was cooled to
<10.degree. C. (ice bath), then diluted with hexanes (800 mL).
The resulting mixture was stirred at ice bath temperature for 4 h,
then filtered. The collected solid was washed with 4:1
hexanes:ethyl acetate (3.times.50 mL) and dried to constant weight
(-30 in. Hg, ambient temperature, 12 h). Recovery=207.5 g of a
white powder. Step C: Preparation of
(S)-(-)-1-(3-chlorophenyl)-1,3-propanediol: ##STR68##
[0613] The compound was prepared as described in Example 7, Step
D.
[0614] The residue was dissolved in methanol (1 mL) and analyzed by
chiral HPLC (see, Example 7; Step B). ee>98%.
Example 10
The Preparation of 1,3-Diols Via Catalytic Asymmetric
Hydrogenation
Step A:
[0615] Beta-ketoester starting material was synthesized as
described in Example 7, step A.
Step B:
[0616] A solution containing beta-ketoester (1 mmol) in either
methanol or ethanol (5-10 mL/mmol ketoester) was degassed through
several pump/vent (N.sub.2) cycles at room temperature. The
degassed solution was moved into a glove bag and under an
atmosphere of N.sub.2 was poured into a stainless steel bomb
containing a stir bar and 1.0 mole % Ru-BINAP catalyst. The bomb
was sealed, removed from the glove bag and purged with H.sub.2
prior to stirring 18-24 h at room temperature and 150 psi H.sub.2.
After venting the hydrogen pressure, the bomb was opened and the
reaction mixture was removed and concentrated. The crude
beta-hydroxyester was used for hydrolysis.
Step C:
[0617] Crude beta-hydroxy ester was hydrolyzed as described in
Example 7, step C.
Step D:
[0618] Optically active beta-hydroxy acid was reduced as described
in Example 7, step D.
Synthesis of Racemic Phosphorylating Agents:
Example 11
General Procedure for the Synthesis of
trans-4-(aryl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinanes
[0619] ##STR69##
Example 11.1
Synthesis of
trans-4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinan-
e
[0620] A solution of 1-(3-chlorophenyl)-1,3-propane diol (25 g, 134
mmol) and triethylamine (62.5 mL, 442 mmol) in THF was added to a
solution of 4-nitrophenyl-phosphorodichloridate (37.7 g, 147 mmol)
in THF at room temperature and the resulting solution was heated at
reflux. After 2 h, TLC indicated complete consumption of the
starting diol and formation of the cis and trans isomers in a 60/40
ratio (HPLC). The clear yellow solution was cooled to 30.degree.
C., sodium 4-nitrophenoxide (56 g, 402 mmol)) was added and the
reaction mixture was heated at reflux. After 90 min. the reddish
reaction mixture was cooled to room temperature and stirred at room
temperature for 2 h then placed in the refrigerator overnight. The
final ratio was determined by HPLC to be 96/4 trans/cis. The
reaction mixture was quenched with a saturated solution of ammonium
chloride and diluted with ethyl acetate. The layers were separated
and the organics were washed 4 times with 0.3 N sodium hydroxide to
remove the nitrophenol, then saturated sodium chloride and dried
over sodium sulfate. The filtered solution was concentrated under
reduced pressure and the resulting solid was recrystallized from
ethyl acetate to give large off white needles (45 g,
mp=115-116.degree. C., purity 98 A %).
[0621] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
cis-isomer 5.6-5.8 (m, 1H), trans-isomer 5.5-5.69 (m, 1H).
[0622] TLC conditions: Merck silica gel 60 F254 plates, 250 .mu.m
thickness; mobile phase=60/40 hexanes/ethyl acetate; R.sub.f:
diol=0.1, cis-phosphate=0.2, trans-phosphate=0.35.
[0623] HPLC conditions: Column=Waters .mu. Bondapack C18
3.9.times.300 mm; mobile phase=40/60 acetonitrile/phosphate buffer
pH 6.2; flow rate=1.4 mL/min; detection=UV @ 270 nm; retention
times in min: cis-isomer=14.46, trans-isomer=16.66,
4-nitrophenol=4.14.
Example 11.2
Synthesis of
trans-4-(3-pyrid-3-yl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0624] Same as Example 11.1
[0625] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.6-5.8 (m, 1H)
Example 11.3
Synthesis of
trans-4-(3,-5-difluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphospho-
rinane
[0626] Same as Example 11.1
[0627] TLC conditions: Merck silica gel 60 F254 plates, 250 .mu.m
thickness; mobile phase=50/50 hexanes/ethyl acetate; R.sub.f:
diol=0.1, cis-phosphate=0.25, trans-phosphate=0.4.
[0628] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.7-5.5 (m, 1H)
Example 11.4
Synthesis of
trans-4-(4-methylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinan-
e
[0629] Same as Example 11.1 starting with
1-(4-methylphenyl)-1,3-propanediol
[0630] TLC: 50/50 hexanes/ethyl acetate; Rf: cis-phosphate=0.25;
trans-phosphate=0.35.
[0631] 1H NMR (CDCl3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.65-5.5 (m, 1H)
Example 11.5
Synthesis of
trans-4-3,5-dimethylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphori-
nane
[0632] Same as Example 11.1 starting with
1-(3,5-dimethylphenyl)-1,3-propanediol
[0633] TLC: 50/50 hexanes/ethyl acetate; Rf: cis-phosphate=0.2;
trans-phosphate=0.3.
[0634] 1H NMR (CDCl3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.6-5.45 (m, 1H)
Example 11.6
Synthesis of
trans-4-(3,5-dichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphor-
inane
[0635] Same as Example 11.1 starting with
1-(3,5-dichlorophenyl)-1,3-propanediol
[0636] TLC: 70/30 hexanes/ethyl acetate; Rf: cis-phosphate=0.3;
trans-phosphate=0.5.
[0637] 1H NMR (CDCl3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.85-5.7 (m, 1H)
Example 11.7
Synthesis of
trans-4-(pyrid-4-yl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0638] Same as Example 11.1 starting with
1-(pyrid-4-yl)-1,3-propanediol
[0639] TLC: 95/5 dichloromethane/ethanol; Rf:
trans-phosphate=0.35.
[0640] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.7-5.55 (m, 1H)
Example 11.8
Synthesis of
trans-4-(3-methoxycarbonylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxapho-
sphorinane
[0641] Same as Example 11.1 starting with
1-(3-methoxycarbonylphenyl)-1,3-propanediol
[0642] TLC: 30/70 hexanes/ethyl acetate; Rf: cis-phosphate=0.5;
trans-phosphate=0.6.
[0643] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.7-5.6 (m, 1H)
Example 11.9
Synthesis of
trans-4-(4-methoxycarbonylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxapho-
sphorinane
[0644] Same as Example 11.1 starting with
1-(4-methoxycarbonylphenyl)-1,3-propanediol
[0645] TLC: 30/70 hexanes/ethyl acetate; Rf: cis-phosphate=0.35;
trans-phosphate=0.5.
[0646] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.7-5.6 (m, 1H)
Example 11.10
Synthesis of
trans-4-(5-bromopyrid-3-yl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphori-
nane
[0647] Same as Example 11.1 starting with
1-(5-bromopyrid-3-yl)-1,3-propanediol
[0648] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.8-5.65 (m, 1H)
Example 11.11
Synthesis of
trans-4-(2,3-dichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphor-
inane
[0649] Same as Example 11.1 starting with
1-(2,3-dichlorophenyl)-1,3-propanediol except that the
isomerization was conducted with 4-nitrophenol and lithium hydride
as in Example 13a.
[0650] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 6-5.9 (m, 1H)
Example 11.12
Synthesis of
trans-4-(2,3,5-trichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosp-
horinane
[0651] Same as Example 11.1 starting with
1-(2,3,5-trichlorophenyl)-1,3-propanediol except that the
isomerization was conducted with 4-nitrophenol and triethylamine as
in Example 13b.
[0652] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.9-5.7 (m, 1H)
Example 11.13
Synthesis of
trans-4-(2-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinan-
e
[0653] Same as Example 11.1 starting with
1-(2-chlorophenyl)-1,3-propanediol except that the isomerization
was conducted with 4-nitrophenol and lithium hydride as in Example
13a.
[0654] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 6-5.9 (m, 1H)
Example 11.14
Synthesis of
trans-4-(3,5-dimethoxyphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphospho-
rinane
[0655] Same as Example 11.1 starting with
1-(3,5-dimethoxyphenyl)-1,3-propanediol except that the
isomerization was conducted with 4-nitrophenol and triethylamine as
in Example 13b.
[0656] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.55-5.45 (m, 1H), 3.3 (s, 6H)
Example 11.15
Synthesis of
trans-4-(2-bromophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0657] Same as Example 11.1 starting with
1-(2-bromophenyl)-1,3-propanediol except that the isomerization was
conducted with 4-nitrophenol and triethylamine as in Example
13a.
[0658] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.95-5.85 (m, 1H)
Example 11.16
Synthesis of
trans-4-(3-bromo-5-ethoxyphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphos-
phorinane
[0659] Same as Example 11.1 starting with
1-(3-bromo-5-ethoxyphenyl)-1,3-propanediol except that the
isomerization was conducted with 4-nitrophenol and triethylamine as
in Example 13b.
[0660] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.9-5.75 (m, 1H), 4.04 (q, 2H), 1.39 (t, 3H).
Example 11.17
Synthesis of
trans-4-(2-trifluoromethylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxapho-
sphorinane
[0661] Same as Example 11.1 starting with
1-(2-trifluoromethylphenyl)-1,3-propanediol except that the
isomerization was conducted with 4-nitrophenol and triethylamine as
in Example 13b.
[0662] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 6-5.75 (m, 1H).
Example 11.18
Synthesis of
trans-4-(4-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinan-
e
[0663] Same as Example 11.1 starting with
1-(4-chlorophenyl)-1,3-propanediol except that the trans-isomer was
isolated from the cis/trans mixture without isomerization.
[0664] TLC: hexanes/ethyl acetate 1/1; Rf: cis-phosphate=0.2;
trans-phosphate=0.6.
[0665] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.6-5.5 (m, 1H).
Example 11.19
Synthesis of
trans-4-(3-methylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinan-
e
[0666] Same as Example 11.1 starting with
1-(3-methylphenyl)-1,3-propanediol except that the trans-isomer was
isolated from the cis/trans mixture without isomerization.
[0667] TLC: hexanes/ethyl acetate 6/4; Rf: cis-phosphate=0.2;
trans-phosphate=0.5.
[0668] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.65-5.5 (m, 1H).
Example 11.20
Synthesis of
trans-4-(4-fluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinan-
es
[0669] Same as Example 11.1 starting with
1-(4-fluorophenyl)-1,3-propanediol except that the trans-isomer was
isolated from the cis/trans mixture without isomerization.
[0670] .sup.1H NMR (DMSO-d.sub.6, Varian Gemini 200 MHz):
C'-proton: trans-isomer 5.78-5.85 (m, 1H).
Example 11.21
Synthesis of
trans-4-(2-fluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinan-
e
[0671] Same as Example 11.1 starting with
1-(2-fluorophenyl)-1,3-propanediol except that the trans-isomer was
isolated from the cis/trans mixture without isomerization.
[0672] .sup.1H NMR (DMSO-d.sub.6, Varian Gemini 200 MHz):
C'-proton: trans-isomer 5.9-6.1 (m, 1H).
Example 11.22
Synthesis of
trans-4-(3-fluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinan-
e
[0673] Same as Example 11.1 starting with
1-(3-fluorophenyl)-1,3-propanediol except that the trans-isomer was
isolated from the cis/trans mixture without isomerization.
[0674] .sup.1H NMR (DMSO-d.sub.6, Varian Gemini 200 MHz):
C'-proton: trans-isomer 5.8-5.9 (m, 1H).
Example 11.23
Synthesis of
trans-4-[4-(4-chlorophenoxy)phenyl]-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxap-
hosphorinane
[0675] Same as Example 11.1 starting with
1-[4-(4-chlorophenoxy)phenyl]-1,3-propanediol except that the
trans-isomer was isolated from the cis/trans mixture without
isomerization.
[0676] .sup.1H NMR (DMSO-d.sub.6, Varian Gemini 200 MHz):
C'-proton: trans-isomer 5.75-5.9 (m, 1H).
Example 11.24
Synthesis of
trans-4-(3-bromophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0677] Same as Example 11.1 starting with
1-(3-bromophenyl)-1,3-propanediol except that the trans-isomer was
isolated from the cis/trans mixture without isomerization.
[0678] TLC: hexanes/ethyl acetate 1/1; Rf: cis-phosphate=0.25;
trans-phosphate=0.5.
[0679] .sup.1H NMR (DMSO-d.sub.6, Varian Gemini 200 MHz):
C'-proton: trans-isomer 5.8-5.95 (m, 1H).
Example 11.25
Synthesis of
trans-4-(3,4-ethylenedioxyphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxapho-
sphorinane
[0680] Same as Example 11.1 starting with
1-(3,4-ethylenedioxyphenyl)-1,3-propanediol except that the
trans-isomer was isolated from the cis/trans mixture without
isomerization.
[0681] TLC: hexanes/ethyl acetate 1/1; Rf: trans-phosphate=0.6.
[0682] .sup.1H NMR (DMSO-d.sub.6, Varian Gemini 200 MHz):
C'-proton: trans-isomer 5.8-5.9 (m, 1H).
Example 11.26
Synthesis of
trans-4-(2-fluoro-4-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxapho-
sphorinane
[0683] Same as Example 11.1 starting with
1-(2-fluoro-4-chlorophenyl)-1,3-propanediol except that the
trans-isomer was isolated from the cis/trans mixture without
isomerization.
[0684] TLC: hexanes/ethyl acetate 1/1; Rf: trans-phosphate=0.7.
[0685] .sup.1H NMR (DMSO-d.sub.6, Varian Gemini 200 MHz):
C'-proton: trans-isomer 5.9-6 (m, 1H).
Example 11.27
Synthesis of
trans-4-(2,6-dichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphor-
inane
[0686] Same as Example 11.1 starting with
1-(2,6-dichlorophenyl)-1,3-propanediol except that the trans-isomer
was isolated from the cis/trans mixture without isomerization.
[0687] TLC: hexanes/ethyl acetate 1/1; Rf:
trans-phosphate=0.65.
[0688] .sup.1H NMR (DMSO-d.sub.6, Varian Gemini 200 MHz):
C'-proton: trans-isomer 6.2-6.4 (m, 1H).
Example 11.28
Synthesis of
trans-4-(2-fluoro-5-methoxyphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaph-
osphorinane
[0689] Same as Example 11.1 starting with
1-(2-fluoro-5-methoxyphenyl)-1,3-propanediol except that the
trans-isomer was isolated from the cis/trans mixture without
isomerization.
[0690] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.8-5.95 (m, 1H), 3.8 (s, 3H).
Example 11.29
Synthesis of
trans-4-(3-fluoro-4-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxapho-
sphorinane
[0691] Same as Example 11.1 starting with
1-(3-fluoro-4-chlorophenyl)-1,3-propanediol except that the
isomerization was conducted with 4-nitrophenol and triethylamine as
in Example 13b.
[0692] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.4-5.6 (m, 1H).
Example 11.30
Synthesis of
trans-4-(3-chloro-4-fluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxapho-
sphorinane
[0693] Same as Example 11.1 starting with
1-(3-chloro-4-fluorophenyl)-1,3-propanediol except that the
isomerization was conducted with 4-nitrophenol and triethylamine as
in Example 13b.
[0694] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.5-5.6 (m, 1H).
Example 11.31
Synthesis of
trans-4-(2-fluoro-5-bromophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphos-
phorinane
[0695] Same as Example 11.1 starting with
1-(2-fluoro-5-bromophenyl)-1,3-propanediol except that the
isomerization was conducted with 4-nitrophenol and triethylamine as
in Example 13b.
[0696] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.8-5.9 (m, 1H).
Example 11.32
Synthesis of
trans-4-(2,3,5,6-tetrafluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxap-
hosphorinane
[0697] Same as Example 11.1 starting with
1-(2,3,5,6-tetrafluorophenyl)-1,3-propanediol except that the
isomerization was conducted with 4-nitrophenol and triethylamine as
in Example 13b.
[0698] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.9-6 (m, 1H).
Example 11.33
Synthesis of
trans-4-(2,3,6-trifluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosp-
horinane
[0699] Same as Example 11.1 starting with
1-(2,3,6-trifluorophenyl)-1,3-propanediol except that the
isomerization was conducted with 4-nitrophenol and triethylamine as
in Example 13b.
[0700] .sup.1H NMR (CDCl.sub.3, Varian Gemini 200 MHz): C'-proton:
trans-isomer 5.9-6 (m, 1H).
Example 11.34
Synthesis of
trans-4(R)-(phenyl)-2-(4-chlorophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0701] Same as Example 11.1 starting with
1(R)-(phenyl)-1,3-propanediol isolated by column without the
isomerization.
[0702] Rf=0.5 (50% EtOAc in Hexanes). mp 90-92.degree. C. Anal
calcd for C.sub.15H.sub.14ClO.sub.4P: C, 55.49; H, 4.35. Found: C,
55.64; H, 3.94.
Example 11.35
Synthesis of
trans-4(R)-(phenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0703] Same as Example 11.1 starting with
1(R)-(phenyl)-1,3-propanediol isolated by column without the
isomerization.
[0704] Rf=0.4 (50% EtOAc in Hexanes). mp 130-131.degree. C. Anal
calcd for C.sub.15H.sub.14NO.sub.6P: C, 53.74; H, 4.21; N, 4.18.
Found: C, 53.86; H, 4.07; N, 4.00.
Example 11.36
Synthesis of
trans-4(S)-(phenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0705] Same as Example 11.1 starting with
1(S)-(phenyl)-1,3-propanediol.
[0706] Rf=0.2 (5% EtOAc in CH.sub.2Cl.sub.2). mp 128-129.degree. C.
Anal calcd for C.sub.15H.sub.14NO.sub.6P: C, 53.74; H, 4.21; N,
4.18. Found: C, 53.69; H, 4.53; N, 4.23.
Example 11.37
Synthesis of
trans-4-(3-trifluoromethylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxapho-
sphorinane
[0707] Same as Example 11.1 starting with
1-(3-trifluoromethylphenyl)-1,3-propanediol.
[0708] Rf=0.32(35% EtOAc in hexanes). mp 78-81.degree. C. Anal
calcd for C.sub.16H.sub.13F.sub.3NO.sub.6P: C, 47.66; H, 3.25; N,
3.47. Found: C, 47.69; H, 3.77; N, 3.52.
Example 11.38
Synthesis of
trans-4-(2,4-dichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphor-
inane
[0709] Same as Example 11.1 starting with
1-(2,4-dichlorophenyl)-1,3-propanediol.
[0710] Rf=0.32(35% EtOAc in hexanes). mp 154-157.degree. C. Anal
calcd for C.sub.15H.sub.12Cl.sub.2NO.sub.6P: C, 44.58; H, 2.99; N,
3.47. Found: C, 44.63; H, 3.07; N, 3.47.
Example 11.39
Synthesis of
trans-4-(3-bromo-4-fluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphos-
phorinane
[0711] Same as Example 11.1 starting with
1-(3-bromo-4-fluorophenyl)-1,3-propanediol. Rf=0.2 (5% EtOAc in
CH.sub.2Cl.sub.2). mp 108.degree. C. Anal calcd for
C.sub.15H.sub.12NO.sub.6BrFP: C, 41.69; H, 2.80; N, 3.24. Found: C,
41.90; H, 2.76; N, 3.05.
Example 11.40
Synthesis of
trans-4-(2-pyridyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0712] Same as Example 11.1 starting with
1-(2-pyridyl)-1,3-propanediol. mp 99-102.degree. C. Anal calcd for
C.sub.14H.sub.13N.sub.2O.sub.6P: C, 50.01; H, 3.90; N, 8.33. Found:
C, 49.84; H, 3.41; N, 8.14.
Example 11.41
Synthesis of
trans-4-(3,4-dichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphor-
inane
[0713] Same as Example 11.1 starting with
1-(3,4-dichlorophenyl)-1,3-propanediol. Rf=0.15 (35% EtOAc in
Hexanes). mp 126-129.degree. C. Anal calcd for
C.sub.15H.sub.12Cl.sub.2NO.sub.6P: C, 44.58; H, 2.99; N, 3.47.
Found: C, 44.71; H, 3.49; N, 3.41.
Example 11.42
Synthesis of
trans-4-(4-tert-butylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphor-
inane
[0714] Same as Example 11.1 starting with
1-(4-tert-butylphenyl)-1,3-propanediol. Rf=0.20 (35% EtOAc in
Hexanes). mp 108-111.degree. C. Anal calcd for
C.sub.19H.sub.22NO.sub.6P: C, 58.31; H, 5.67; N, 3.58. Found: C,
58.04; H, 5.67; N, 3.55.
Example 11.43
Synthesis of
trans-4-(3-thiophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0715] Same as Example 11.1 starting with
1-(3-thiophenyl)-1,3-propanediol. mp 94-96.degree. C. Anal calcd
for C.sub.13H.sub.12NO.sub.6PS: C, 45.75; H, 3.54; N, 4.10. Found:
C, 45.65; H, 3.21; N, 4.24.
Example 11.44
Synthesis of
trans-4-(3-furanyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0716] Same as Example 11.1 starting with
1-(3-furanyl)-1,3-propanediol. mp 108-111.degree. C. Anal calcd for
C.sub.13H.sub.12NO.sub.7P: C, 48.01; H, 3.72; N, 4.31. Found: C,
48.06; H, 3.61; N, 4.26.
Example 11.45
Synthesis of
trans-4-(2-bromo-5-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphos-
phorinane
[0717] Same as Example 11.1 starting with
1-(2-bromo-5-chlorophenyl)-1,3-propanediol. Rf=0.20 (5% MeOH in
CH.sub.2Cl.sub.2). mp 105-106.degree. C. Anal calcd for
C.sub.15H.sub.12NO.sub.6BrClP: C, 40.16; H, 2.70; N, 3.12. Found:
C, 39.97; H, 2.86; N, 3.06.
Example 11.46
Synthesis of
trans-4-(2,5-difluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphor-
inane
[0718] Same as Example 11.1 starting with
1-(2,5-difluorophenyl)-1,3-propanediol. Rf=0.50 (50% EtOAc in
Hexanes). mp 120-122.degree. C. Anal calcd for
C.sub.15H.sub.12F.sub.2NO.sub.6P: C, 48.53; H, 3.26; N, 3.77.
Found: C, 48.46; H, 3.52; N, 3.87.
Example 11.47
Synthesis of
trans-4-(2,4-difluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphor-
inane
[0719] Same as Example 11.1 starting with
1-(2,4-difluorophenyl)-1,3-propanediol. Rf=0.50 (50% EtOAc in
Hexanes). mp 85-87.degree. C. Anal calcd for
C.sub.15H.sub.12F.sub.2NO.sub.6P: C, 48.53; H, 3.26; N, 3.77.
Found: C, 48.82; H, 3.55; N, 3.84.
Example 11.48
Synthesis of
trans-4-cis-6-(diphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinan-
e
[0720] Same as Example 11.1 starting with
trans-1,3-diphenyl-1,3-propanediol (Yamamura, H., Araki, S.,
Tetrahedron, 1997, 53, 46, 15685-15690) without equilibration.
Rf=0.29 (35% EtOAc in Hexanes). mp 118-121.degree. C. Anal calcd
for C.sub.21H.sub.18NO.sub.6P: C, 61.32; H, 4.41; N, 3.41. Found:
C, 60.94; H, 4.44; N, 3.53.
Example 11.49
Synthesis of
trans-4-trans-6-(diphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorin-
ane
[0721] Same as Example 11.1 starting with
cis-1,3-diphenyl-1,3-propanediol (Yamamura, H., Araki, S.,
Tetrahedron, 1997, 53, 46, 15685-15690) without equilibration.
Rf=0.65 (5% EtOAc in CH.sub.2Cl.sub.2). mp 144-147.degree. C. Anal
calcd for C.sub.21H.sub.18NO.sub.6P: C, 61.32; H, 4.41; N, 3.41.
Found: C, 61.21; H, 4.58; N, 3.36.
Example 11.50
Synthesis of
cis-4-cis-6-(diphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0722] Same as Example 11.1 starting with
cis-1,3-diphenyl-1,3-propanediol (Yamamura, H., Araki, S.,
Tetrahedron, 1997, 53, 46, 15685-15690) without equilibration.
Rf=0.3 (5% EtOAc in CH.sub.2Cl.sub.2). mp 135-138.degree. C. Anal
calcd for C.sub.21H.sub.18NO.sub.6P: C, 61.32; H, 4.41; N, 3.41.
Found: C, 61.29; H, 4.77; N, 3.46.
Example 11.51
Synthesis of
cis-4-cis-5-(diphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0723] Same as Example 11.1 starting with
cis-1,2-diphenyl-1,3-propanediol (Kristersson, P, Lindquist, K.,
Acta Chem. Scand. B 1980, 34, 3, 213-234) without equilibration.
Rf=0.35 (5% EtOAc in CH.sub.2Cl.sub.2). mp 136-139.degree. C. Anal
calcd for C.sub.21H.sub.18NO.sub.6P: C, 61.32; H, 4.41; N, 3.41.
Found: C, 60.95; H, 4.41; N, 3.82.
Example 11.52
Synthesis of
trans-4-trans-5-(diphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorin-
ane
[0724] Same as Example 11.1 starting with
cis-1,2-diphenyl-1,3-propanediol (Kristersson, P, Lindquist, K.,
Acta Chem. Scand. B 1980, 34, 3, 213-234) without equilibration.
Rf=0.65 (5% EtOAc in CH.sub.2Cl.sub.2). mp 176-178.degree. C. Anal
calcd for C.sub.21H.sub.18NO.sub.6P: C, 61.32; H, 4.41; N, 3.41.
Found: C, 61.09; H, 4.46; N, 3.80.
Example 11.53
Synthesis of
trans-4-4-dimethyl-6-(phenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphospho-
rinane
Step A:
[0725] To a solution of diisopropylamine (58.4 g, 577 mmol) in dry
ether (500 mL) at -78.degree. C. under nitrogen was added n-BuLi
(215 mL, 2.5 M in hexane, 538 mmol) over 30 min. The reaction was
stirred for 10 min before addition of ethyl acetate (55 mL, 558
mmol) over a period 30 min. Freshly distilled benzaldehyde (47 mL,
443 mmol) in ether (50 mL) was slowly added over 30 min and the
mixture was allowed to warm to room temperature. The reaction was
quenched with saturated ammonium chloride (150 mL) at 0.degree. C.
The organic layer was washed, dried (anhydrous Na.sub.2SO.sub.4)
and concentrated to give the crude addition product.
Step B:
[0726] To a solution of crude condensation product (10.6 g, 54.6
mmol) in dry ether at -78.degree. C. was added MeMgBr (60 mL, 3.0 M
in THF, 180 mmol). The mixture was allowed to warm to room
temperature and stirred overnight. The reaction was quenched with
ammonium chloride (50 mL) at 0.degree. C. and diluted with EtOAc
(350 mL). The organic layer was washed, dried (anhydrous
Na.sub.2SO.sub.4) and concentrated. The crude product was purified
by column chromatography (0-10% EtOAc in CH.sub.2Cl.sub.2) to give
3,3-dimethyl-1-phenyl-1,3-propanediol (7 g) as a pale yellow
oil.
Step C:
[0727] Same as Example 11.1 starting with
3,3-dimethyl-1-phenyl-1,3-propanediol without equilibration.
Rf=0.18 (35% EtOAc in hexanes). mp 131-133.degree. C. Anal calcd
for C.sub.17H.sub.18NO.sub.6P: C, 56.20; H, 4.99; N, 3.86. Found:
C, 56.00; H, 5.03; N, 3.86.
Example 11.54
Synthesis of
cis-4-(3-chlorophenyl)-cis-5-methoxy-(-2-(4-nitrophenoxy)-2-oxo-1,3,2-dio-
xaphosphorinane and
trans-4-(3-chlorophenyl)-cis-5-methoxy-(-2-(4-nitrophenoxy)-2-oxo-1,3,2-d-
ioxaphosphorinane (11.55)
Step A:
[0728] To a solution of lithium diisopropylamide (356 mmol) in THF
(500 mL) at -78.degree. C. was slowly added 2-methoxy-methyl
acetate (38.8 mL, 392 mmol) via an addition funnel. The reaction
was stirred at -78.degree. C. for 30 min before
3-chlorobenzaldehyde (20.1 mL, 178 mmol) was added. The reaction
was allowed to warm to room temperature and quenched with saturated
aq NH.sub.4Cl (500 mL). The mixture was extracted with EtOAc
(3.times.200 mL) and the combined organic extracts were washed with
water and dried (anhydrous Na.sub.2SO.sub.4). The crude product was
purified by column chromatography (5-50% EtOAc in hexanes) to yield
3-(3-chlorophenyl)-3-hydroxy-2-methoxy-methyl proprionate (39 g) as
pale yellow oil.
Step B:
[0729] To a solution of the ester (39 g, 159 mmol) obtained from
step A in ethanol (500 mL) was added sodiumborohydride (6.2 g, 159
mmol) in three portions, over 10 min. The reaction was refluxed for
3 h and the ethanol was evaporated under reduced pressure. The
residue was dissolved in EtOAc (500 mL), washed with water and
dried (anhydrous Na.sub.2SO.sub.4). The crude product was purified
by column chromatography (1-5% MeOH--CH.sub.2Cl.sub.2) to give the
diol (28 g) as colorless oil.
Step C:
[0730] To a solution of diol (28 g, 129 mmol) in acetone (250 mL)
was added trimethyl orthoformate (10 mL) followed by
p-toluenesulfonic acid (500 mg, 2.64 mmol) and the reaction was
heated to reflux overnight. The reaction was cooled to room
temperature and the acetone was removed under vacuum. The residue
was dissolved in ethyl acetate and washed with NaHCO.sub.3, water
and dried (anhydrous Na.sub.2SO.sub.4). The ketals were separated
by column chromatography (5-10% EtOAc in hexanes) to give 1,2-cis
(7.26 g) and 1,2-trans ketal (0.9 g) diastereomers.
Step D:
[0731] The 1,2-cis ketal (4.5 g, 17.5 mmol) was dissolved in 70% aq
TFA (10 mL) and allowed to react overnight at room temperature. The
reaction was diluted with acetonitrile (30 mL) and volatiles were
removed under reduced pressure. The residue was dissolved in EtOAc
(300 mL) and the organic layer was washed with saturated aq
NaHCO.sub.3, water and dried (anhydrous Na.sub.2SO.sub.4). The
crude product was purified by column chromatography (1-5%
MeOH--CH.sub.2Cl.sub.2) to give 1,2-cis diol diastereomer (3.5
g).
[0732] The 1,2-trans ketal diastereomer was also hydrolyzed
following the above procedure to give 1,2-trans-diol
diastereomer.
Step E:
[0733] 1,2-cis-diol diastereomer was subjected to phosphorylation
using the procedure described in Example 11.1 without equilibration
to give the following two isomers.
[0734] 11.54: Rf=0.57 (5% EtOAc in CH.sub.2Cl.sub.2). mp
110-112.degree. C. Anal calcd for C.sub.16H.sub.15NO.sub.7PCl: C,
48.08; H, 3.78; N, 3.50. Found: C, 48.35; H, 3.56; N, 3.69.
[0735] 11.55: Rf=0.34 (5% EtOAc in CH.sub.2Cl.sub.2). mp
131-134.degree. C. Anal calcd for C.sub.16H.sub.15NO.sub.7PCl
0.0.3H.sub.2O: C, 47.44; H, 3.88; N, 3.46. Found: C, 47.23; H,
4.01; N, 3.46.
Example 12
General Procedure for the Synthesis of
trans-4-(aryl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinanes
Using Phosphorus Oxychloride
[0736] Phosphorus oxychloride (3.4 mL, 36.3 mmol) was added to a
solution of 1-(3-chlorophenyl)-1,3-propanediol in dichloromethane
at 0.degree. C. followed by triethylamine (10.2 mL, 73 mmol). After
2 h, sodium 4-nitrophenoxide (10.63 g, 66 mmol) was added to the
solution of cis/trans phosphorochloridate reagent and the orange
reaction mixture was heated at reflux for 1 h. The cooled solution
was partitioned with ethyl acetate and a saturated solution of
ammonium chloride. The organics were separated and dried over
sodium sulfate, filtered and concentrated under reduced pressure.
The residue was taken up in THF, sodium 4-nitrophenoxide (10.63 g,
66 mmol) was added and the orange reaction mixture was heated to
reflux for 3 h (HPLC, 95/5 trans/cis). The cooled solution was
partitioned with ethyl acetate and a saturated solution of ammonium
chloride. The organics were separated and washed with 0.3 N
solution of sodium hydroxide and brine, dried over sodium sulfate
and concentrated under reduced pressure. Recrystallization from
ethyl acetate as in Example 10 gave the phosphate reagent.
Example 13
Procedures for the Enrichment in Trans-isomer of a Cis/Trans
Mixture of
4-(aryl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0737] A cis/trans mixture of
4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinanes
was prepared as in Example 11, except that the cis and trans
isomers were separated by column chromatography prior to the
addition of 4-nitrophenol.
[0738]
Cis-4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphospho-
rinane was isomerized to the trans isomer by adding a solution of
the cis-isomer to a solution of 4-nitrophenoxide prepared with the
following bases.
Example 13a
[0739] Lithium hydride (19.4 mg, 2.44 mmol) was added to a solution
of 4-nitrophenol in THF at room temperature. The yellow solution
was stirred at room temperature for 30 min. A solution of
cis-4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
(300 mg, 0.813 mmol) in THF was added to the solution of lithium
4-nitrophenoxide. The orange reaction mixture was stirred a room
temperature. After 5 h the ratio was 92.9/5.4 trans/cis (HPLC
determination).
Example 13b
[0740] Same as above using triethylamine instead of lithium
hydride. After 20 h the trans/cis ratio was 90.8/5.3.
Example 13c
[0741] Same as above using DBU instead of lithium hydride. After 3
h the trans/cis ratio was 90.8/5.3.
Synthesis of Enantioenriched Phosphorylating Agents
Example 14
General Procedure for the Synthesis of Enantioenriched
trans-4-(aryl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinanes
[0742] ##STR70##
Example 14a
Synthesis of
(+)-(4R)-trans-4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxapho-
sphorinane
[0743] A solution of (+)-(R)-1-(3-chlorophenyl)-1,3-propanediol (3
g, 16.1 mmol) and triethylamine (6.03 ml, 59.6 mmol) in THF (80 mL)
was added dropwise to a solution of
4-nitrophenoxyphosphorodichloridate (7.63 g, 29.8 mmol) in 150 mL
of THF at 0.degree. C. After about 2 h, the starting diol was
consumed, with the formation of two isomeric
4-nitophenylphosphates, and additional triethylamine (8.31 mL)
followed by of 4-nitrophenol (8.29 g, 59.6 mmol) were added. The
reaction mixture was stirred overnight. The solvent was evaporated
under reduced pressure and the residue was partitioned between
ethyl acetate and water. The organic phase was washed (0.4 M NaOH,
water and sat'd NaCl solution) and dried over MgSO.sub.4.
Concentration and chromatography of the residue using 30% ethyl
acetate in hexanes yielded 4.213 g of the desired product.
[0744] HNMR (200 MHz, CDCl.sub.3): 8.26 (2H, d, J=9.7 Hz), 7.2-7.5
(6H, m), 5.56 (1H, apparent d, J=11.7 Hz), 4.4-4.7 (2H, m), 2.2-2.6
(1H, m), 2.0-2.2 (1H, m).
[0745] mp: 114-115.degree. C. [.alpha.].sub.D=+91.71. Elemental
Analysis: Calculated for C.sub.15H.sub.13NO.sub.6ClP: C, 48.73; H,
3.54; N, 3.79. Found: C, 48.44; H, 3.20; N, 3.65
Example 14b
Synthesis of
(-)-(4S)-trans-4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxapho-
sphorinane
[0746] ##STR71##
[0747] In a similar manner, from 3.116 g of
(-)-(S)-1-(3-chlorophenyl)-1,3-propane diol was obtained 4.492 g of
the desired phosphate.
[0748] HNMR (200 MHz, CDCl.sub.3): 8.26 (2H, d, J=9.7 Hz), 7.2-7.5
(6H, m), 5.56 (1H, apparent d, J=11.7 Hz), 4.4-4.7 (2H, m), 2.2-2.6
(1H, m), 2.02-2.2 (1H, m).
[0749] mp: 114-115.degree. C. [.alpha.].sub.D=-91.71. Elemental
Analysis: Calculated for C.sub.15H.sub.13NO.sub.6ClP: C, 48.73; H,
3.54; N, 3.79. Found: C, 48.61; H, 3.36; N, 3.66.
Example 14c
Synthesis of
(-)-(4S)-trans-phenyl-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane
[0750] Same as Example 11.1 starting with
S-(-)-1-phenyl-1,3-propanediol except that the isomerization was
conducted with 4-nitrophenol and triethylamine as in Example
13b.
[0751] TLC: hexanes/ethyl acetate 4/1); Rf=0.4
[0752] .sup.1H NMR (DMSO-d.sub.6, Varian Gemini 300 MHz):
C'-proton: trans-isomer 5.85-5.75 (m, 1H).
Example 15
General Procedures for Maintaining Enantiomeric Excess During
Synthesis of Enantioenriched Phosphorylating Reagent
Example 15a
Synthesis of
(-)-(4S)-trans-(pyrid-4-yl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphori-
nane
[0753] ##STR72##
[0754] A 12 L round bottom flask equipped with an overhead stirrer
and nitrogen inlet was charged with
(S)-(-)-1-(pyrid-4-yl)-1,3-propanediol (1.2 kg, 7.83 mol) and
pyridine (6 L) The mixture was vigorously stirred at room
temperature for 0.5 h until all the solids had dissolved.
Meanwhile, a 22 L, 3-neck flask was equipped with an overhead
stirrer, thermocouple, cooling bath, and nitrogen inlet. This
vessel was charged with 4-nitrophenyl phosphorodichloridate (2.01
kg, 7.83 mol) and pyridine (6 L). The resulting mixture was cooled
to 3.3.degree. C. After the diol was completely dissolved (0.5 h),
triethylamine (190 mL, 1.36 mol) was added and the slightly cloudy,
yellow-brown solution was transferred in portions to a 2 L addition
funnel on the 22 L flask. The solution was added to the cold
phosphorodichloridate solution over 3.25 h. After the addition was
complete, the cooling bath was drained and stirring was continued
for 3 h. During this time, a 50 L, 3-neck flask was equipped with
an overhead stirrer, thermocouple, addition funnel, cooling bath
(ice water) and nitrogen inlet. This flask was then charged with
sodium hydride (180 g, 4.5 mol) and THF (1 L) and the addition
funnel was charged with a solution of 4-nitrophenol (817 g, 5.87
mol) in THF (1 L). The nitrophenol solution was slowly added to the
cold suspension of sodium hydride. After the addition was complete,
the resulting bright orange suspension was stirred at room
temperature for 1 h. After the diol-dichloridate reaction was
judged complete the dark suspension was subjected to vacuum
filtration. The glassware and filter cake (triethylamine-HCl) were
rinsed with THF (1 L) and the combined filtrate and rinse were
poured into the orange, sodium 4-nitrophenoxide suspension. The
resulting mixture was then heated at 40.degree. C. for 3.5 h at
which time the heating mantle was turned off and the reaction was
stirred an additional 11 h at room temperature. The crude reaction
mixture was concentrated on a rotary evaporator at 45-50.degree. C.
at reduced pressure (oil pump). The resulting thick, black, foamy
tar was dissolved in 1.5 M aq HCl (12 L) and ethyl acetate (8 L).
The mixture was transferred to a 12.5-gallon separatory funnel,
stirred 10 min, and the phases separated. The ethyl acetate layer
was washed with an additional 1.3 L of 1.5 M aq HCl. To the
combined aqueous layers was added dichloromethane (8 L) and the
vigorously stirred mixture was carefully neutralized with solid
sodium bicarbonate. The layers were separated and the aqueous layer
was extracted with dichloromethane (8 L). The combined organic
layers were dried over magnesium sulfate (600 g) and filtered. The
solution was concentrated on a rotary evaporator until most of the
solvent was removed and a thick suspension resulted. 2-Propanol (5
L) was added and evaporation continued until 4 L of distillate were
collected. 2-Propanol (3 L) was added and evaporation continued
until 3 L of distillate were collected. The thick slurry was
diluted with 2-propanol (2 L) and the mixture stirred with cooling
(ice bath) for 1 h. The solid was collected by filtration, washed
with 2-propanol (2 L), and dried in a vacuum oven (-30 in. Hg,
55.degree. C., 18 h) to a constant weight of 1.86 kg.
[0755] mp 140-142.degree. C.
[0756] Specific Rotation=-80.350 (c=1.0, MeOH); ee=99+% trans
[0757] HPLC conditions:
[0758] Column: Chiralpak AD, 0.46.times.25 cm; mobile phase=50:50,
2-propanol:hexane, isocratic; flow rate=1.0 mL/min; injection
volume=10 .mu.L UV detection at 254 nm.
[0759] The cis/trans equilibration was monitored by HPLC. Stopped
at 92% trans, 6.6% cis, r.t.=trans isomer 6.9 min. and cis isomer
10.9 min.
[0760] .sup.1HNMR (DMSO-d.sub.6): .delta.=2.23-2.29 (m, 2H),
4.56-4.71 (m, 2H), 5.88-5.95 (m, 1H), 7.44 (d, 2H, J=5.8 Hz), 7.59
(d, 2H, J=9.2 Hz), 8.34 (d, 2H, J=9.4 Hz), 8.63 (d, 2H J=5.8
Hz)
Example 15b
Synthesis of
(-)-(4S)-(-)-(pyrid-4-yl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorina-
ne
[0761] A 1 liter 3-neck round bottom flask was equipped with a
mechanical stirrer, addition funnel, a thermometer and a N.sub.2
inlet. The flask is charged with
S-(-)-1-(pyrid-4-yl)-propane-1,3-diol (25 g, 163.4 mmol) and ethyl
acetate (250 mL) and the resulting suspension was treated slowly
with a 4N HCl solution in dioxane (43 mL, 176 mmol) over a period
of 15 min. After stirring for 30 min at room temperature,
4-nitrophenylphosphorodichloridate (41.81 g, 163.4 mmol) was added
as a solid as quickly as possible under a positive flow of N2. The
internal temperature of the reaction mixture was adjusted to
-10.degree. C. with the help of a dry ice-acetone cooling bath. A
solution of triethylamine (79 mL, 572 mmol) in ethyl acetate (100
mL) was added maintaining the reaction temperature at
<-5.degree. C. Thirty minutes after the complete addition of the
triethylamine solution, the cooling bath was removed and the
reaction mixture was stirred at room temperature for 1 h. The
reaction mixture was filtered to remove triethylamine-hydrochloride
salt, which is washed with ethyl acetate (3.times.30 mL) until the
filtrate shows only faint absorption. The filtrate was evaporated
down to a volume of 150-175 mL under reduced pressure.
4-nitrophenol (7.5 g, 54.3 mmol) and triethylamine (9 mL) were
added to the concentrated solution and the resulting orange
reaction mixture was stirred at room temperature for 24 h. The
solid formed in the reaction mixture was collected by filtration,
washed with ethyl acetate (2.times.25 mL) and methyl t-butyl ether
(25 mL) and dried under vacuum at 55.degree. C. to give 31.96 g of
the desired product. Same analytical data as Example 14a.
Example 16
Preparation of Prodrugs of 2'-C-beta-methyl-7-deazaadenosine Via
trans-phosphate Addition
16.1:
4-amino-7-(cis-5'-O-[4-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorina-
n-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0762] ##STR73## Step A:
[0763] To a solution of
4-amino-7-(2-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
(US2002-0147160A1, WO 02/057827) (10 g, 0.356 mol) in anhydrous
acetone (145 mL) and anhydrous DMF (35 mL) were added p-toluene
sulfonic acid monohydrate (33.8 g, 0.18 moles) and triethyl
orthoformate (31.2 mL, 28.5 moles) at room temperature. The
reaction was warmed to .about.80.degree. C. and allowed to stir for
3 h under nitrogen. The mixture was evaporated under reduced
pressure. The oily residue was purified by column chromatography
(5% MeOH in CH.sub.2Cl.sub.2) to give the isopropylidene derivative
(8.6 g) as a white solid.
Step B:
[0764] To a solution of
2',3'-O-isopropylidene-4-amino-7-(2-C-methyl-beta-D-ribofuranosyl)-7H-pyr-
rolo[2,3-d]pyrimidine (0.094 g, 0.29 mmol) in DMF (1.5 mL) was
added t-butyl magnesium chloride and stirred under nitrogen for 30
min. The reaction mixture was then cooled to -55.degree. C. and the
phosphorylating agent (whose preparation is described in example
11.1) (0.13 g, 0.35 mmol) in DMF (1.5 mL) was added dropwise. The
reaction was allowed to warm to room temperature and stirred under
nitrogen for 2 h. The mixture was evaporated under reduced pressure
and purified by chromatography (5% MeOH in CH.sub.2Cl.sub.2) to
yield 0.070 g of the 2',3'-O-isopropylidene protected prodrug as a
yellow solid.
Step C:
[0765] The prodrug (0.15 g, 0.27 mmol) obtained from the above step
was dissolved in pre-cooled 75% TFA/H2O (20 mL) and allowed to stir
at 0.degree. C. for 2 h. The reaction mixture was evaporated under
reduced pressure. The crude product was purified by flash
chromatography (1% aq.NH.sub.4OH in 10% MeOH in CH.sub.2Cl.sub.2)
to give 0.142 g of the title compound as an off-white solid.
[0766] R.sub.f=0.40 (10% MeOH in CH.sub.2Cl.sub.2). mp
138-141.degree. C. Anal calcd for
C.sub.21H.sub.24ClN.sub.4O.sub.7P.0.4 CH.sub.2Cl.sub.2: C, 47.18;
H, 4.59; N, 10.28. Found: C, 46.97; H, 4.59; N, 10.11.
[0767] The following examples were synthesized as described in
steps A-C of example 16.1, utilizing the phosphorylating agents of
examples 1-15.
16.2:
4-amino-7-(cis-5'-O-[4-(2,5-difluorophenyl)-2-oxo-1,3,2-dioxaphospho-
rinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0768] ##STR74##
[0769] R.sub.f=0.35 (10% MeOH in CH.sub.2Cl.sub.2). mp
145-148.degree. C. Anal Calcd for
C.sub.21H.sub.23N.sub.4O.sub.7F.sub.2P.1.35H.sub.2O.1.0
CF.sub.3CO.sub.2H: C, 42.45; H, 4.14; N, 8.62. Found: C, 42.18; H,
3.77; N, 8.42.
16.3:
4-amino-7-(cis-5'-O-[4-(3-chloro-4-fluorophenyl)-2-oxo-1,3,2-dioxaph-
osphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimi-
dine
[0770] ##STR75##
[0771] R.sub.f=0.30 (10% MeOH in CH.sub.2Cl.sub.2). mp
128-130.degree. C. Anal Calcd for
C.sub.21H.sub.23N.sub.4O.sub.7FClP.2H.sub.2O.1.9CF.sub.3CO.sub.2H:
C, 38.11; H, 3.73; N, 7.17. Found: C, 38.04; H, 3.28; N, 7.02.
16.4:
4-amino-7-(cis-5'-O-[6,6-dimethyl-4-phenyl-2-oxo-1,3,2-dioxaphosphor-
inan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0772] ##STR76##
[0773] R.sub.f=0.40 (10% MeOH in CH.sub.2Cl.sub.2). mp
140-142.degree. C. Anal Calcd for
C.sub.23H.sub.29N.sub.4O.sub.7P.1H.sub.2O.0.4 CF.sub.3CO.sub.2H: C,
50.32; N, 5.57; N, 9.86. Found: C, 50.38; H, 5.12; N, 9.96.
16.5:
4-amino-7-(cis-5'-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphospho-
rinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0774] ##STR77##
[0775] R.sub.f=0.45 (10% MeOH in CH.sub.2Cl.sub.2). mp
135-138.degree. C. Anal Calcd for
C.sub.21H.sub.24ClN.sub.4O.sub.7P.0.2H.sub.2O.0.4 CH.sub.2Cl.sub.2:
C, 46.87; H, 4.63; N, 10.22. Found: C, 47.02; H, 4.25; N, 9.99.
16.6:
4-amino-7-(cis-5'-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphospho-
rinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
Methanesulfonic Acid Salt
[0776] ##STR78##
[0777] R.sub.f=0.45 (10% MeOH in CH.sub.2Cl.sub.2). mp
125-128.degree. C. Anal Calcd for
C.sub.21H.sub.24N.sub.4O.sub.7ClP.1.6
CH.sub.3SO.sub.3H.1.0H.sub.2O: C, 39.76; H, 4.78; N, 8.21; S, 7.52.
Found: C, 39.39; H, 4.30; N, 8.30; S, 7.96.
16.7:
4-amino-7-(cis-5'-O-[4-(S)-(pyridin-4-yl)-2-oxo-1,3,2-dioxaphosphori-
nan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0778] ##STR79##
[0779] R.sub.f=0.40 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
mp 183-185.degree. C. Anal Calcd for
C.sub.20H.sub.24N.sub.5O.sub.7P. 1.6H.sub.2O: C, 47.45; H, 5.42; N,
13.83. Found: C, 47.78; H, 5.47; N, 13.77.
16.8:
4-amino-7-(cis-5'-O-[4-(3-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorina-
n-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0780] ##STR80##
[0781] R.sub.f=0.15 (10% MeOH in CH.sub.2Cl.sub.2). Anal Calcd for
C.sub.21H.sub.24FN.sub.4O.sub.7P. 0.3 H.sub.2O: C, 50.46; H, 4.96;
N, 11.21. Found: C, 50.63; H, 5.35; N, 10.94.
16.9:
4-amino-7-(cis-5'-O-[4-(3-bromophenyl)-2-oxo-1,3,2-dioxaphosphorinan-
-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0782] ##STR81##
[0783] Rf=0.48 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH). Anal
Calcd for C.sub.21H.sub.24BrN.sub.4O.sub.7P. 0.5 H.sub.2O: C,
44.70; H, 4.47; N, 9.93. Found: C, 44.58; H, 4.52; N, 9.56.
16.10:
4-amino-7-(cis-5'-O-[4-(2-bromophenyl)-2-oxo-1,3,2-dioxaphosphorina-
n-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0784] ##STR82##
[0785] R.sub.f=0.15 (10% MeOH in CH.sub.2Cl.sub.2). mp
132-135.degree. C. Anal Calcd for
C.sub.21H.sub.24BrN.sub.4O.sub.7P. 0.5 H.sub.2O: C, 44.7; H, 4.47;
N, 9.93. Found: C, 44.73; H, 4.69; N, 9.82.
16.11:
4-amino-7-(cis-5'-O-[4-(5-bromopyridin-3-yl)-2-oxo-1,3,2-dioxaphosp-
horinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-
e
[0786] ##STR83##
[0787] R.sub.f=0.35 (10% MeOH in EtOAc) mp 132-135.degree. C. Anal
Calcd for C.sub.20H.sub.23N.sub.5O.sub.7BrP. 0.5 H.sub.2O. 0.5
EtOAc: C, 43.36; H, 4.63; N, 11.49.
[0788] Found: C, 43.37; H, 4.80; N, 11.16.
16.12:
4-amino-7-(cis-5'-O-[4-(S)-phenyl-2-oxo-1,3,2-dioxaphosphorinan-2-y-
l]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0789] ##STR84##
[0790] R.sub.f=0.42 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
mp 115-118.degree. C. Anal Calcd for
C.sub.21H.sub.25N.sub.4O.sub.7P. 0.4 EtOAc. 1.0 H.sub.2O: C, 51.25;
H, 5.75; N, 10.58. Found: C, 51.07; H, 5.88; N, 10.35.
16.13:
4-amino-7-(cis-5'-O-[4,5-cis-diphenyl-2-oxo-1,3,2-dioxaphosphorinan-
-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
Trifluoroacetic Acid Salt
[0791] ##STR85##
[0792] R.sub.f=0.45 (10% MeOH in CH.sub.2Cl.sub.2). mp
174-177.degree. C. Anal Calcd for
C.sub.29H.sub.30F.sub.3N.sub.4O.sub.9P.1.75 H.sub.2O: C, 49.90; H,
4.48; N, 8.03. Found: C, 49.68; H, 4.82; N, 8.1.
16.14:
4-amino-7-(cis-5'-O-[4-(2-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorin-
an-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0793] ##STR86##
[0794] R.sub.f=0.48 (10% MeOH in CH.sub.2Cl.sub.2). mp
187-190.degree. C. Anal Calcd for
C.sub.21H.sub.24ClN.sub.4O.sub.7P. H.sub.2O. 0.2 DMF: C, 47.72; H,
5.05; N, 10.77. Found: C, 47.66; H, 5.02; N, 10.96.
16.15:
4-amino-7-(cis-5'-O-[4-(2-fluoro-5-bromophenyl)-2-oxo-1,3,2-dioxaph-
osphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimi-
dine
[0795] ##STR87##
[0796] R.sub.f=0.48 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
Anal Calcd for C.sub.21H.sub.23BrFN.sub.4O.sub.7P. 1.3H2O: C,42.27;
H, 4.32; N, 9.39. Found: C, 42.26; H, 4.03; N, 9.36.
16.16:
4-amino-7-(cis-5'-O-[4,6-cis-diphenyl-2-oxo-1,3,2-dioxaphosphorinan-
-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
Trifluoroacetic Acid Salt
[0797] ##STR88##
[0798] R.sub.f=0.20 (10% MeOH in CH.sub.2Cl.sub.2). mp
140-143.degree. C. Anal Calcd for
C.sub.27H.sub.29N.sub.4O.sub.7P.1.25 H.sub.2O.CF.sub.3CO.sub.2H: C,
50.55; H, 4.75; N, 8.13. Found: C, 50.25; H, 4.88; N, 7.99.
16.17:
4-amino-7-(cis-5'-O-[4(3,5-bis-trifluoromethylpheny)-2-oxo-1,3,2-di-
oxaphosphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]p-
yrimidine
[0799] ##STR89##
[0800] R.sub.f=0.15 (10% MeOH in CH.sub.2Cl.sub.2). mp
130-134.degree. C. Anal Calcd for C.sub.23H.sub.23N.sub.4O.sub.7P.
0.6 H.sub.2O: C, 44.33; H, 3.91; N, 8.99. Found: C, 44.29; H, 4.13;
N, 8.98.
16.18:
4-amino-7-(trans-5'-O-[4,6-cis-diphenyl-2-oxo-1,3,2-dioxaphosphorin-
an-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
Trifluoroacetic Acid Salt
[0801] ##STR90##
[0802] Rf=0.48 (15% MeOH in CH.sub.2C.sub.2-1% NH.sub.4OH).
mp>220.degree. C. Anal Calcd for
C.sub.27H.sub.29N.sub.4O.sub.7P.0.9 H.sub.2O: C, 57.02; H, 5.46; N,
9.85. Found: C, 57.55; H, 5.97; N, 9.88.
16.19:
4-amino-7-(trans-5'-O-[4-(3-bromo-pyridin-4-yl)-2-oxo-1,3,2-dioxaph-
osphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimi-
dine
[0803] ##STR91##
[0804] R.sub.f=0.3 (10% MeOH in EtOAc). mp 116-120.degree. C. Anal
Calcd for C.sub.20H.sub.23N.sub.5O.sub.7BrP.1 H2O. 0.6 EtOAc: C,
42.90; H, 4.79; N, 11.17 Found: C, 42.90; H, 4.42; N, 10.82.
16.20:
4-amino-7-(trans-5'-O-[4-(2,4-dichlorophenyl)-2-oxo-1,3,2-dioxaphos-
phorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidi-
ne
[0805] ##STR92##
[0806] R.sub.f=0.15 (10% MeOH in CH.sub.2Cl.sub.2). mp
184-188.degree. C. Anal Calcd for
C.sub.22H.sub.24F.sub.3N.sub.4O.sub.7P. 0.6 H.sub.2O: C, 47.59; H,
4.57; N, 10.09. Found: C, 47.46; H, 4.96; N, 10.10.
16.21:
4-amino-7-(trans-5'-O-[4-(3-trifluoromethylphenyl)-2-oxo-1,3,2-diox-
aphosphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyr-
imidine
[0807] ##STR93##
[0808] R.sub.f=0.15 (10% MeOH in CH.sub.2Cl.sub.2). mp
120-124.degree. C. Anal Calcd for
C.sub.21H.sub.23Cl.sub.2N.sub.4O.sub.7P.0.5 H.sub.2O: C, 45.50; H,
4.36; N, 10.11. Found: C, 45.32; H, 4.58; N, 10.26.
16.22:
4-amino-7-(trans-5'-O-[4,5-cis-diphenyl-2-oxo-1,3,2-dioxaphosphorin-
an-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0809] ##STR94##
[0810] R.sub.f=0.75 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
mp 160-163.degree. C. Anal Calcd for
C.sub.27H.sub.29N.sub.4O.sub.7P.1.2 H.sub.2O: C, 56.48; H, 5.51; N,
9.76. Found: C, 56.34; H, 5.75; N, 9.71.
16.23:
4-amino-7-(cis-5'-O-[cis-(5-methoxy-4-phenyl)2-oxo-1,3,2-dioxaphosp-
horinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-
e Trifluoroacetic Acid Salt
[0811] ##STR95##
[0812] R.sub.f=0.25 (10% MeOH in CH.sub.2Cl.sub.2). mp
116-120.degree. C. Anal Calcd for
C.sub.22H.sub.26N.sub.4O.sub.8PCl.1.75 H.sub.2O.1.5
CF.sub.3CO.sub.2H: C, 40.39; H, 4.20; N, 7.54. Found: C, 39.95; H,
3.85; N, 7.38.
16.24:
4-amino-7-(cis-5'-O-[trans-(5-methoxy-4-phenyl)2-oxo-1,3,2-dioxapho-
sphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimid-
ine Trifluoroacetic Acid Salt
[0813] ##STR96##
[0814] R.sub.f=0.30 (10% MeOH in CH.sub.2Cl.sub.2). mp
140-143.degree. C. Anal Calcd for
C.sub.22H.sub.26N.sub.4O.sub.8PCl.2.5 H.sub.2O.2.2
CF.sub.3CO.sub.2H: C, 37.89; H, 4.00; N, 6.70. Found: C, 37.73; H,
3.61; N, 6.85.
16.25:
4-amino-7-(cis-5'-O-[4-(2-bromo-5-chlorophenyl)-2-oxo-1,3,2-dioxaph-
osphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimi-
dine
[0815] ##STR97##
[0816] R.sub.f=0.3 (10% MeOH in CH.sub.2Cl.sub.2). mp
193-196.degree. C. Anal Calcd for
C.sub.21H.sub.23N.sub.4O.sub.7PClBr.1.75 H.sub.2O.1
CF.sub.3CO.sub.2H: C, 37.57; H, 3.77; N, 7.62. Found: C, 37.20; H,
3.49; N, 7.36.
16.26:
4-amino-7-(cis-5'-O-[4-(3,5-dichlorophenyl)-2-oxo-1,3,2-dioxaphosph-
orinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0817] ##STR98##
[0818] R.sub.f=0.3 (10% MeOH in CH.sub.2Cl.sub.2). mp
182-185.degree. C. Anal Calcd for
C.sub.21H.sub.23N.sub.4O.sub.7Cl.sub.2P.0.3 MeOH0.0.5 H.sub.2O: C,
45.37; H, 4.50; N, 9.93. Found: C, 45.36; H, 4.18; N, 9.58.
16.27:
4-amino-7-(cis-5'-O-[4-(3,5-difluorophenyl)-2-oxo-1,3,2-dioxaphosph-
orinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0819] ##STR99##
[0820] R.sub.f=0.35 (10% MeOH in CH.sub.2Cl.sub.2). mp
135-140.degree. C. Anal Calcd for
C.sub.21H.sub.23N.sub.4O.sub.7F.sub.2P.1.0 H.sub.2O: C, 47.55; H,
4.75; N, 10.56. Found: C, 47.29; H, 4.51; N, 10.28.
16.28:
4-amino-7-(cis-5'-O-[4-(R)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosph-
orinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0821] ##STR100##
[0822] Rf=0.45 (10% MeOH in CH.sub.2Cl.sub.2). mp 126-128.degree.
C. Anal Calcd for C.sub.21H.sub.24ClN.sub.4O.sub.7P.1.0 H.sub.2O:
C, 47.69; H, 4.96; N, 1059. Found: C, 47.31; H, 4.77; N, 10.3.
16.29:
4-amino-7-(cis-5'-O-[4-(2-trifluoromethylphenyl)-2-oxo-1,3,2-dioxap-
hosphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrim-
idine
[0823] ##STR101##
[0824] Rf=0.5 (10% MeOH in CH.sub.2Cl.sub.2). mp 115-120.degree. C.
Anal Calcd for C.sub.22H.sub.24F.sub.3N.sub.4O.sub.7P.1.0
H.sub.2O.1.0 CF.sub.3CO.sub.2H: C, 42.61; H, 4.02; N, 8.28. Found:
C, 42.78; H, 4.07; N, 8.27.
16.30:
4-amino-7-(cis-5'-O-[4-(R)-(pyridin-4-yl)-2-oxo-1,3,2-dioxaphosphor-
inan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0825] ##STR102##
[0826] R.sub.f=0.3 (20% MeOH in EtOAc). mp 132-136.degree. C. Anal
Calcd for C.sub.20H.sub.24N.sub.5O.sub.7P.0.03 H.sub.2O.0.7
CH.sub.2Cl.sub.2: C, 46.52; H, 4.79; N, 13.14. Found: C, 46.13; H,
4.39; N, 13.50.
16.31:
4-amino-7-(cis-5'-O-[4-(3-bromo-4-fluoro-phenyl)-2-oxo-1,3,2-dioxap-
hosphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrim-
idine
[0827] ##STR103##
[0828] R.sub.f=0.35 (10% MeOH in EtOAc). mp 122-125.degree. C. Anal
Calcd for C.sub.21H.sub.23N.sub.4O.sub.7FBrP.0.2 CF.sub.3CO.sub.2H:
C, 43.12; H, 3.92; N, 9.40. Found: C, 42.82; H, 3.76; N, 9.57.
16.32:
4-amino-7-(cis-5'-O-[4-(pyridin-3-yl)-2-oxo-1,3,2-dioxaphosphorinan-
-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0829] ##STR104##
[0830] R.sub.f=0.30 (10% MeOH in EtOAc). mp 134-138.degree. C. Anal
Calcd for C.sub.20H.sub.24N.sub.5O.sub.7P.1.5 H.sub.2O: C, 47.62;
H, 5.40; N, 13.88. Found: C, 47.89; H, 5.08; N, 13.97.
16.33:
4-amino-7-(cis-5'-O-[4-(pyridin-2-yl)-2-oxo-1,3,2-dioxaphosphorinan-
-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
Trifluoroacetic Acid Salt
[0831] ##STR105##
[0832] R.sub.f=0.50 (10% MeOH in CH.sub.2Cl.sub.2). mp
88-90.degree. C. Anal Calcd for C.sub.20H.sub.24N.sub.5O.sub.7P.2.3
H.sub.2O.1.3 CF.sub.3CO.sub.2H: C, 40.69; H, 4.52; N, 10.50. Found:
C, 40.38; H, 4.86; N, 10.90.
16.34:
4-amino-7-(cis-5'-O-[4-(R)-(phenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-
-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0833] ##STR106##
[0834] R.sub.f=0.30 (10% MeOH in CH.sub.2Cl.sub.2). mp
177-180.degree. C. Anal Calcd for C.sub.21H.sub.25N.sub.4O.sub.7P.
0.1 EtOAc. 0.2 CF.sub.3CO.sub.2H: C, 51.54; H, 5.16; N, 11.03.
Found: C, 51.92; H, 4.78; N, 10.75.
16.35:
4-amino-7-(cis-5'-O-[4-(4-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorin-
an-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
Trifluoroacetic Acid Salt
[0835] ##STR107##
[0836] R.sub.f=0.45 (10% MeOH in CH.sub.2Cl.sub.2). mp
182-184.degree. C. Anal Calcd for
C.sub.21H.sub.24N.sub.4O.sub.7ClP.2.0 H.sub.2O.2.9
CF.sub.3CO.sub.2H: C, 36.68; H, 3.55; N, 6.38. Found: C, 36.33; H,
3.35; N, 6.44.
16.36:
4-amino-7-(cis-5'-O-[4-(2,3-difluorophenyl)-2-oxo-1,3,2-dioxaphosph-
orinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
Trifluoroacetic Acid Salt
[0837] ##STR108##
[0838] R.sub.f=0.5 (10% MeOH in CH.sub.2Cl.sub.2). mp
177-180.degree. C. Anal Calcd for
C.sub.21H.sub.23F.sub.2N.sub.4O.sub.7P.1.9
H.sub.2O.1.1CF.sub.3CO.sub.2H: C, 41.46; H, 4.18; N, 8.34. Found:
C, 42.07; H, 4.02; N, 8.68.
16.37:
4-amino-7-(cis-5'-O-[4-(2-fluoro-5-methoxyphenyl)-2-oxo-1,3,2-dioxa-
phosphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyri-
midine Trifluoroacetic Acid Salt
[0839] ##STR109##
[0840] R.sub.f=0.4 (10% MeOH in CH.sub.2Cl.sub.2). mp 80-85.degree.
C. Anal Calcd for C.sub.22H.sub.26N.sub.4O.sub.8FP.0.4 H.sub.2O.2.0
CF.sub.3CO.sub.2H: C, 41.11; H, 3.82; N, 7.37. Found: C, 41.13; H,
3.50; N, 7.54.
16.38:
4-amino-7-(cis-5'-O-[4-(2-chloro-4-fluorophenyl)-2-oxo-1,3,2-dioxap-
hosphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrim-
idine Trifluoroacetic Acid Salt
[0841] ##STR110##
[0842] Rf=0.46 (15% MeOH in CH.sub.2Cl.sub.2). mp 138-141.degree.
C. Anal Calcd for C.sub.21H.sub.23ClFN.sub.4O.sub.7P. 0.3 H.sub.2O.
0.9 CF.sub.3CO.sub.2H: C, 43.00; H, 3.88; N, 8.80. Found: C, 42.73;
H, 4.21; N, 8.55.
16.39:
4-amino-7-(cis-5'-O-[4-(2-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorin-
an-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0843] ##STR111##
[0844] Rf=0.48 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH). mp
101-103.degree. C. Anal Calcd for C.sub.21H.sub.24FN.sub.4O.sub.7P.
1.5 H.sub.2O: C, 48.37; H, 5.22; N, 10.74. Found: C, 48.70; H,
5.47; N, 10.43.
16.40:
4-amino-7-(cis-5'-O-[4-(2-cyanophenyl)-2-oxo-1,3,2-dioxaphosphorina-
n-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0845] ##STR112##
[0846] Rf=0.42 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH). Anal
Calcd for C.sub.22H.sub.24N.sub.5O.sub.7P. 2 H.sub.2O. 0.1
CF.sub.3CO.sub.2H: C, 48.58; H, 5.16; N, 12.76. Found: C, 48.86; H,
5.51; N, 12.70.
16.41:
4-amino-7-(cis-5'-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosph-
orinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
Trifluoroacetic Acid Salt
[0847] ##STR113##
[0848] R.sub.f=0.45 (10% MeOH in CH.sub.2Cl.sub.2). mp
145-148.degree. C. Anal Calcd for
C.sub.21H.sub.24N.sub.4O.sub.7PCl.0.7 CH.sub.2Cl.sub.2.1.2
CF.sub.3CO.sub.2H: C, 40.93; H, 3.79; N, 7.92; F, 9.67.
[0849] Found: C, 40.43; H, 3.77; N, 8.22; F, 9.47.
16.42:
4-amino-7-(cis-5'-O-[4-phenyl-5,6-tetramethylene-2-oxo-1,3,2-dioxap-
hosphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrim-
idine
[0850] ##STR114##
[0851] R.sub.f=0.24 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
mp 110-113.degree. C. Anal Calcd for
C.sub.25H.sub.31N.sub.4O.sub.7P. 2.0 H.sub.2O: C, 53.00; H, 6.23;
N, 9.89. Found: C, 53.03; H, 5.93; N, 9.91.
16.43:
4-amino-7-(cis-5'-O-[4-(3-cyanophenyl)-2-oxo-1,3,2-dioxaphosphorina-
n-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0852] ##STR115##
[0853] R.sub.f=0.51 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
mp 157-160.degree. C. Anal Calcd for
C.sub.22H.sub.24N.sub.5O.sub.7P. 2.5H.sub.2O: C, 48.35; H, 5.35; N,
12.82. Found: C, 48.50; H, 5.72; N, 12.77.
Example 17
Preparation of Prodrugs of 2'-C-beta-methyl-7-deazaguanosine Via
trans-phosphate Addition
[0854] The parent nucleoside
2-amino-7-(2-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(-
3H)-one was synthesized as described in US2002-0147160A1 and WO
02/057827.
[0855] The nucleoside was converted to corresponding prodrug
following the procedures as in steps A, B and C of Example 16.
[0856] The following examples were synthesized as described steps
A-C.
17.1:
2-amino-7-(cis-5'-O-[4-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorina-
n-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0857] ##STR116##
[0858] R.sub.f=0.30 (10% MeOH in CH.sub.2Cl.sub.2). Anal calcd for
C.sub.21H.sub.24ClN.sub.4O.sub.8P.1.2 CF.sub.3CO.sub.2NH.sub.4.1.0
CF.sub.3CO.sub.2H: C, 38.22; H, 3.76; N, 9.13. Found: C, 37.93; N,
3.80; N, 9.40.
17.2:
2-amino-7-(cis-5'-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphospho-
rinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0859] ##STR117##
[0860] R.sub.f=0.15 (10% MeOH in CH.sub.2Cl.sub.2). mp 175.degree.
C. Anal Calcd for C.sub.21H.sub.24ClN.sub.4O.sub.8P.0.5H.sub.2O: C,
47.07; H, 4.70; N, 10.46. Found: C, 46.73; H, 4.90; N, 10.16.
17.3:
2-amino-7-(cis-5'-O-[4-(5-bromo-2-fluorophenyl)-2-oxo-1,3,2-dioxapho-
sphorinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0861] ##STR118##
[0862] R.sub.f=0.41 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
Anal Calcd for C.sub.21H.sub.23BrFN.sub.4O.sub.8P. 0.5 H.sub.2O.
0.2 CF.sub.3CO.sub.2H: C, 41.38; H, 3.93; N, 9.02. Found: C, 41.60;
H, 4.32; N, 8.77.
17.4:
2-amino-7-(cis-5'-O-[4-(3-bromophenyl)-2-oxo-1,3,2-dioxaphosphorinan-
-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
Trifluoroacetic Acid Salt
[0863] ##STR119##
[0864] R.sub.f=0.38 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
mp 142-145.degree. C. Anal Calcd for
C.sub.21H.sub.24N.sub.4O.sub.8P. 0.7H.sub.2O. 0.9
CF.sub.3CO.sub.2H: C, 39.89; H, 3.86; N, 8.16. Found: C, 39.53; H,
3.65; N, 8.43.
17.5:
2-amino-7-(cis-5'-O-[4-(3-Chloro-4-fluorophenyl)-2-oxo-1,3,2-dioxaph-
osphorinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0865] ##STR120##
[0866] R.sub.f=0.45 (20% MeOH in CH.sub.2Cl.sub.2. Anal Calcd for
C.sub.21H.sub.23N.sub.4O.sub.8FClP. 1.4 H.sub.2O: C, 44.24, H,
4.78; N, 9.83. Found: C, 43.77; H, 4.78; N, 10.31.
17.6:
2-amino-7-(cis-5'-O-[4-(2,5-difluorophenyl)-2-oxo-1,3,2-dioxaphospho-
rinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0867] ##STR121##
[0868] R.sub.f=0.35 (20% MeOH in CH.sub.2Cl.sub.2). mp
170-173.degree. C. Anal Calcd for
C.sub.21H.sub.23F.sub.2N.sub.4O.sub.8P.2.0 H.sub.2O.0.4
CF.sub.3CO.sub.2NH.sub.4: C, 42.45; H, 4.67; N, 9.99. Found: C,
42.28; H, 4.76 N, 9.96.
17.7:
2-amino-7-(cis-5'-O-[4-(2-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorina-
n-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0869] ##STR122##
[0870] R.sub.f=0.25 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
Anal Calcd for C.sub.21H.sub.24ClN.sub.4O.sub.8P. 1.25 H.sub.2O.
0.2 CF.sub.3CO.sub.2H: C, 44.92; H, 4.70; N, 9.79. Found: C, 44.93;
H, 5.09; N, 10.08.
17.8:
2-amino-7-(cis-5'-O-[4-(pyridin-2-yl)-2-oxo-1,3,2-dioxaphosphorinan--
2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-o- ne
Trifluoroacetic Acid Salt
[0871] ##STR123##
[0872] R.sub.f=0.4 (15% MeOH in CH.sub.2Cl.sub.2). mp
180-190.degree. C. Anal Calcd for
C.sub.20H.sub.24N.sub.5O.sub.8P.1.3 CF.sub.3CO.sub.2H.0.3
CH.sub.2Cl.sub.2: C, 41.23; H, 3.91; N, 10.50. Found: C, 40.96; H,
3.46; N, 11.05.
17.9:
2-amino-7-(cis-5'-O-[4-(2-trifluoromethylphenyl)-2-oxo-1,3,2-dioxaph-
osphorinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
Trifluoroacetic Acid Salt
[0873] ##STR124##
[0874] R.sub.f=0.4 (10% MeOH in CH.sub.2Cl.sub.2). mp
185-188.degree. C. Anal Calcd for
C.sub.22H.sub.24N.sub.4O.sub.8F.sub.3P.0.8 CF.sub.3CO.sub.2H: C,
43.50; H, 3.84; N, 8.60. Found: C, 43.55; H, 3.97; N, 8.98.
17.10:
2-amino-7-(cis-5'-O-[4-(R)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosph-
orinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
Trifluoroacetic Acid Salt
[0875] ##STR125##
[0876] Rf=0.50 (15% MeOH in CH.sub.2Cl.sub.2). mp 170-180.degree.
C.
17.11:
2-amino-7-(cis-5'-O-[4-(3,5-difluorophenyl)-2-oxo-1,3,2-dioxaphosph-
orinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
Trifluoroacetic Acid Salt
[0877] ##STR126##
[0878] R.sub.f=0.30 (10% MeOH in CH.sub.2Cl.sub.2) mp
182-185.degree. C. Anal Calcd for
C.sub.21H.sub.23N.sub.4O.sub.8F.sub.2P.0.3 EtOAc. 0.2
CF.sub.3CO.sub.2H: C, 46.99; H, 4.47; N, 9.70. Found: C, 47.26; H,
4.32; N, 9.46.
17.12:
2-amino-7-(cis-5'-O-[4-(3,5-dichlorophenyl)-2-oxo-1,3,2-dioxaphosph-
orinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0879] ##STR127##
[0880] R.sub.f=0.35 (10% MeOH in CH.sub.2Cl.sub.2). mp
177-180.degree. C. Anal Calcd for
C.sub.21H.sub.23N.sub.4O.sub.8C.sub.12P.0.1 EtOAc 0.0.2
CF.sub.3CO.sub.2H. C, 44.16; H, 4.08; N, 9.45. Found: C, 44.33; H,
4.44; N, 9.18.
17.13:
2-amino-7-(cis-5'-O-[4-(S)-(pyridin-4-yl)-2-oxo-1,3,2-dioxaphosphor-
inan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0881] ##STR128##
[0882] R.sub.f=0.21 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
mp 138-141.degree. C. Anal Calcd for
C.sub.20H.sub.24N.sub.5O.sub.8P. 2.2 H.sub.2O: C, 45.07; H, 5.33;
N, 13.14. Found: C, 45.12; H, 5.40; N, 12.89.
17.14:
2-amino-7-(cis-5'-O-[4-(3-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorin-
an-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0883] ##STR129##
[0884] R.sub.f=0.25 (10% MeOH in CH.sub.2Cl.sub.2). mp 170.degree.
C. Anal Calcd for C.sub.21H.sub.24FN.sub.4O.sub.8P.1.5 H.sub.2O: C,
46.93; H, 5.06; N,10.42. Found: C, 46.92; H, 5.12; N, 10.44.
17.15:
2-amino-7-(cis-5'-O-[4-(3-bromo-4-fluoro-phenyl)-2-oxo-1,3,2-dioxap-
hosphorinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0885] ##STR130##
[0886] R.sub.f=0.25 (10% MeOH in CH.sub.2Cl.sub.2). mp
175-179.degree. C. Anal Calcd for
C.sub.21H.sub.23BrFN.sub.4O.sub.8P. 0.5 H.sub.2O. 0.5 EtOAc: C,
43.01; H, 4.39; N, 8.72. Found: C, 43.03; H, 4.49; N, 8.49.
17.16:
2-amino-7-(cis-5'-O-[4-(R)-phenyl-2-oxo-1,3,2-dioxaphosphorinan-2-y-
l]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0887] ##STR131##
[0888] Rf=0.30 (10% MeOH in CH.sub.2Cl.sub.2) mp 128-133.degree. C.
Anal Calcd for C.sub.21H.sub.25N.sub.4O.sub.8P. 1.1 H.sub.2O.0.3
CF.sub.3CO.sub.2H: C, 47.48; H, 5.07; N, 10.25. Found: C, 47.61; H,
5.36; N, 9.91.
17.17:
2-amino-7-(cis-5'-O-[4,5-cis-diphenyl-2-oxo-1,3,2-dioxaphosphorinan-
-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
Trifluoroacetic Acid Salt
[0889] ##STR132##
[0890] R.sub.f=0.45 (20% MeOH in CH.sub.2Cl.sub.2). mp
187-190.degree. C. Anal Calcd for C.sub.27H.sub.29N.sub.4O.sub.8P.2
H.sub.2O.1.3 CF.sub.3CO.sub.2H: C, 47.23; H, 4.59; N, 7.44. Found:
C, 46.83; H, 4.33; N, 7.31.
17.18:
2-amino-7-(cis-5'-O-[6,6-dimethyl-4-phenyl-2-oxo-1,3,2-dioxaphospho-
rinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
Trifluoroacetic Acid Salt
[0891] ##STR133##
[0892] R.sub.f=0.40 (20% MeOH in CH.sub.2Cl.sub.2). mp
192-194.degree. C. Anal Calcd for
C.sub.23H.sub.29N.sub.4O.sub.8P.2.0 H.sub.2O.1.0 CF.sub.3CO.sub.2H:
C, 44.78; H, 5.11; N, 8.36. Found: C, 44.40; H, 4.67; N, 8.22.
17.19:
2-amino-7-(cis-5'-O-[cis-(5-methoxy-4-phenyl)-2-oxo-1,3,2-dioxaphos-
phorinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0893] ##STR134##
[0894] R.sub.f=0.30 (20% MeOH in CH.sub.2Cl.sub.2). mp
148-151.degree. C. Anal Calcd for
C.sub.22H.sub.26N.sub.4O.sub.9ClP.1.0 H.sub.2O: C, 45.96; H, 4.91;
N, 9.75. Found: C, 46.03; H, 4.80; N, 9.64.
17.20:
2-amino-7-(cis-5'-O-[4-(2,3-difluorophenyl)-2-oxo-1,3,2-dioxaphosph-
orinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
Trifluoroacetic Acid Salt
[0895] ##STR135##
[0896] R.sub.f=0.5 (10% MeOH in CH.sub.2Cl.sub.2). mp
215-220.degree. C. Anal Calcd for
C.sub.21H.sub.23N.sub.4O.sub.8F.sub.2P.1.0 H.sub.2O.1.0
CF.sub.3CO.sub.2H: C, 41.83; H, 3.97; N, 8.48. Found: C, 41.70; H,
3.77; N, 8.50.
17.21:
2-amino-7-(cis-5'-O-[4-(2-bromophenyl)-2-oxo-1,3,2-dioxaphosphorina-
n-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0897] ##STR136##
[0898] R.sub.f=0.15 (10% MeOH in CH.sub.2Cl.sub.2). mp 180.degree.
C. Anal Calcd for C.sub.21H.sub.24BrN.sub.4O.sub.8P.1.1 H.sub.2O:
C, 42.67; H, 4.47; N, 9.48. Found: C, 42.51; H, 4.60; N, 9.58.
17.22:
2-amino-7-(cis-5'-O-[4-(3,4-dichlorophenyl)-2-oxo-1,3,2-dioxaphosph-
orinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0899] ##STR137##
[0900] R.sub.f=0.30 (10% MeOH in CH.sub.2Cl.sub.2). mp
192-195.degree. C. Anal Calcd for
C.sub.21H.sub.23N.sub.4O.sub.8Cl.sub.2P.0.2 CF.sub.3CO.sub.2H. 0.2
EtOAc: C, 44.31; H, 4.15; N, 9.31. Found: C, 44.40; H, 3.94; N,
9.21.
17.23:
2-amino-7-(cis-5'-O-[4-(3,5-bis-(trifluoromethylphenyl)-2-oxo-1,3,2-
-dioxaphosphorinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0901] ##STR138##
[0902] R.sub.f=0.15 (10% MeOH in CH.sub.2Cl.sub.2) mp
155-175.degree. C. Anal Calcd for
C.sub.23H.sub.23F.sub.6N.sub.4O.sub.8P.0.6 H.sub.2O: C, 43.22; H,
3.82; N, 8.76. Found: C, 43.08; H, 4.03; N, 8.94.
17.24:
2-amino-7-(cis-5'-O-[4-(3-trifluoromethylphenyl)-2-oxo-1,3,2-dioxap-
hosphorinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0903] ##STR139##
[0904] R.sub.f=0.15 (10% MeOH in CH.sub.2Cl.sub.2). mp
145-165.degree. C. Anal Calcd for
C.sub.22H.sub.24F.sub.3N.sub.4O.sub.8P.1 H.sub.2O: C, 45.68; H,
4.53; N, 9.69. Found: C, 45.31; H, 4.88; N, 9.71.
17.25:
2-amino-7-(cis-5'-O-[4-(2,4-dichlorophenyl)-2-oxo-1,3,2-dioxaphosph-
orinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0905] ##STR140##
[0906] R.sub.f=0.15 (10% MeOH in CH.sub.2Cl.sub.2) mp 175.degree.
C. Anal Calcd for
C.sub.21H.sub.23C.sub.12N.sub.4O.sub.8P.1H.sub.2O: C, 43.54; H,
4.35; N, 9.67. Found: C, 43.32; H, 4.35; N, 9.55.
17.26:
2-amino-7-(cis-5'-O-[4-(5-bromo-pyridin-3-yl)-2-oxo-1,3,2-dioxaphos-
phorinan-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0907] ##STR141##
[0908] R.sub.f=0.3 (10% MeOH in CH.sub.2Cl.sub.2) mp
185-189.degree. C. Anal Calcd for C.sub.20H.sub.23N.sub.5O.sub.8BrP
0.1.5 CF.sub.3CO.sub.2H: C, 37.16; H, 3.32; N, 9.42. Found: C,
37.23; H, 3.44; N, 9.33.
17.27:
2-amino-7-(cis-5'-O-[4-(pyridin-3-yl)-2-oxo-1,3,2-dioxaphosphorinan-
-2-yl]-2'-C
methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one
[0909] ##STR142##
[0910] R.sub.f=0.15 (10% MeOH in CH.sub.2Cl.sub.2); Anal Calcd for
C.sub.20H.sub.24N.sub.5O.sub.8P.1 H.sub.2O.0.4 EtOAc: C, 47.46; H,
5.38; N, 12.81. Found: C, 47.40; H, 5.17; N, 12.78.
Example 18
5'-O-[4-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2'-C-methylad-
enosine
[0911] ##STR143##
[0912] 2'-C-methyl adenosine was made as described in
WO01/90121.
Step A:
[0913] General procedure for synthesis of cyclic phosphoramidites
from substituted diols:
[0914] To a solution of commercially available diisopropyl
phosphoramidous dichloride (1 mmol) in THF (5 mL) was added
1,3-diol (1 mmol) and triethylamine (4 mmol) in THF (5 mL) at
-78.degree. C. over 30 min. The reaction was slowly warmed to room
temperature and left stirring overnight. Reaction mixture was
filtered to remove salts and filtrate was concentrated to give
crude product. Silica gel column chromatography provided pure
cyclic diisopropyl phosphoramidite of 1,3-diol.
Step B:
[0915] General procedure for nucleoside-cyclic phosphoramidite
coupling and oxidation:
[0916] (J. Org. Chem., 1996, 61, 7996)
[0917] To a solution of nucleoside (1 mmol) and cyclic
phosphoramidite (1 mmol) in DMF (10 mL) was added benzimidazolium
triflate (1 mmol). The reaction was stirred for 30 min at room
temperature. The mixture was cooled to -40.degree. C. before
addition of t-butylhydro peroxide (2 mmol) and left at room
temperature overnight. Concentration under reduced pressure and
chromatography of crude product resulted in pure cyclic propyl
prodrug.
[0918] R.sub.f=0.46 (12% MeOH in CH.sub.2Cl.sub.2). mp 153.degree.
C. Anal calcd for C.sub.20H.sub.23ClN.sub.5O.sub.7P: C, 46.93; H,
4.53; N, 13.63. Found: C, 47.06; H, 4.36; N, 13.68.
Example 19
cis-5'-O-[4-(3-Chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2'-C-meth-
yl-guanosine
[0919] ##STR144##
[0920] 2'-C-Methyl guanosine was made as described in
WO01/90121.
[0921] The nucleoside was converted to corresponding prodrug
following the procedures as in steps A, B and C of Example 16.
[0922] R.sub.f=0.35 (25% MeOH in CH.sub.2Cl.sub.2).
mp>230.degree. C. Anal calcd for
C.sub.20H.sub.23ClN.sub.5O.sub.8P: C, 45.51; H, 4.39; N, 13.27.
Found: C, 45.89; H, 4.44; N, 13.23.
Example 20
cis-5'-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2'-C--
beta-methyl-guanosine
[0923] ##STR145##
[0924] The compound was synthesized in a similar sequence as
Example 19 using the phosphorylating agent whose preparation is
described in Example 14.
[0925] R.sub.f=0.35 (20% MeOH in CH.sub.2Cl.sub.2).
mp>180.degree. C. Anal calcd for
C.sub.20H.sub.23N.sub.5O.sub.8ClP.1.0 H.sub.2O.0.8
CF.sub.3CO.sub.2H: C,40.72; H, 4.08; N, 10.99. Found: C, 40.43; N,
4.41; N, 11.34.
Example 21
Preparation of Prodrugs of 2'-C-beta-methyl-adenosine Via
trans-phosphate Addition
[0926] 2'-C-methyl adenosine was made as described in
WO01/90121.
[0927] The nucleoside was converted to corresponding prodrug
following the procedures as in steps A, B and C of Example 16.
[0928] trans-phosphorylating agents utilized in step B are
synthesized by the procedures as described in examples 1-15.
21.1:
cis-5'-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-
-2'-C-beta-methyl-adenosine Trifluoroacetic Acid Salt
[0929] ##STR146##
[0930] R.sub.f=0.3 (5% MeOH in EtOAc). mp 125-128.degree. C. Anal
calcd for C.sub.20H.sub.23ClN.sub.5O.sub.7P.1.7 CF.sub.3CO.sub.2H:
C, 39.83; H, 3.53; N, 9.92. Found: C, 39.52; H, 3.46; N, 10.21.
21.2:
cis-5'-O-[4-(3-cyanophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2'-C-
-beta-methyl-adenosine
[0931] ##STR147##
[0932] R.sub.f=0.43 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
mp 153-156.degree. C. Anal calcd for
C.sub.21H.sub.23N.sub.6O.sub.7P.1.1 H.sub.2O: C, 48.30; H, 4.86; N,
16.09. Found: C, 48.53; H, 5.11; N, 15.75.
21.3:
cis-5'-O-[4-(2,5difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]--
2'-C-beta-methyl-adenosine
[0933] ##STR148##
[0934] R.sub.f=0.60 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
mp 75-78.degree. C. Anal calcd for
C.sub.20H.sub.22F.sub.2N.sub.5O.sub.7P.0.3 CH.sub.2Cl.sub.2: C,
45.25; H, 4.23; N, 13.00. Found: C, 45.07; H, 3.94; N, 12.69.
21.4:
cis-5'-O-[4-(3,5-difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-
-2'-C-beta-methyl-adenosine
[0935] ##STR149##
[0936] R.sub.f=0.65 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
mp 120-123.degree. C. Anal calcd for
C.sub.20H.sub.22F.sub.2N.sub.5O.sub.7P.1.5 H.sub.2O.0.1
C.sub.6H.sub.14: C, 45.07; H, 4.85; N, 12.76. Found: C, 45.04; H,
5.25; N, 12.59.
21.5:
cis-5'-O-[4-(S)-(pyridin-4-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2-
'-C-beta-methyl-adenosine
[0937] ##STR150##
[0938] R.sub.f=0.55 (15% MeOH in CH.sub.2Cl.sub.2-1% NH.sub.4OH).
Anal calcd for C.sub.19H.sub.23N.sub.6O.sub.7P.2.5H.sub.2O: C,
43.60; H, 5.39; N, 16.06. Found: C, 43.35; H, 5.54; N, 16.05.
21.6:
cis-5'-O-[4-(3-bromophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2'-C-
-beta-methyl adenosine
[0939] ##STR151##
[0940] R.sub.f=0.5 (10% MeOH in CH.sub.2Cl.sub.2). mp
108-110.degree. C. Anal calcd for
C.sub.20H.sub.23N.sub.5O.sub.7BrP.1.5 H.sub.2O.0.4
CF.sub.3CO.sub.2H: C, 39.72; H, 4.23; N, 11.14. Found: C, 39.44; H,
4.55; N, 11.18.
21.7:
cis-5'-O-[4-(pyridin-2-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2'-C--
beta-methyl-adenosine Trifluoroacetic Acid Salt
[0941] ##STR152##
[0942] R.sub.f=0.4 (10% MeOH in CH.sub.2Cl.sub.2). mp
118-120.degree. C. Anal calcd for
C.sub.19H.sub.23N.sub.6O.sub.7P.2.0 H.sub.2O.1.0 CF.sub.3CO.sub.2H:
C, 40.14; H, 4.49; N, 13.37. Found: C, 40.36; H, 4.92; N,
13.63.
21.8:
cis-5'-O-[4-(4-methylsulfonylphenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-
-yl]-2'-C-beta-methyl-adenosine Trifluoroacetic Acid Salt
[0943] ##STR153##
[0944] R.sub.f=0.3 (10% MeOH in CH.sub.2Cl.sub.2). mp
185-187.degree. C. Anal calcd for
C.sub.21H.sub.26N.sub.5O.sub.9PS.0.6 H.sub.2O.0.6
CF.sub.3CO.sub.2H: C, 42.01; H, 4.41; N, 11.03. Found: C, 41.93; H,
4.73; N, 10.97.
21.9:
cis-5'-O-[4-(pyridine-3-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl-2'-C--
beta-methyl-adenosine
[0945] ##STR154##
[0946] R.sub.f=0.2 (10% MeOH in EtOAc). mp 137-140.degree. C. Anal
calcd for C.sub.19H.sub.23N.sub.6O.sub.7P.1.5 H.sub.2O.0.4 EtOAc.
C, 45.76; H, 15.54; N, 5.44. Found: C, 45.88; H, 15.19; N,
5.09.
Example 22
General Procedure for Preparation of 3'-acetyl Prodrugs of
2'-C-beta-methyl-7-deazaadenosine Cyclic Prodrugs
[0947] To a solution of 5'-substituted cyclic propyl prodrug (0.3
mmol) in pyridine (3 mL) was added acetic anhydride (0.6 mL) at
0.degree. C. The reaction was left at 0.degree. C. for 18 h. Excess
acetic anhydride was quenched with ethanol (3 mL). The mixture was
concentrated and azeotroped with additional ethanol (2.times.5 mL).
The crude residue was chromatographed to get pure monoacetylated
compound as a solid.
22.1:
4-amino-7-(3'-acetyl-cis-5'-O-[4-(S)-(pyridin-4-yl)-2-oxo-1,3,2-diox-
aphosphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyr-
imidine
[0948] ##STR155##
[0949] R.sub.f=0.35 (15% MeOH in CH.sub.2Cl.sub.2). mp
182-185.degree. C. Anal calcd for
C.sub.22H.sub.26N.sub.5O.sub.8P.1.5 H.sub.2O.0.2 CH.sub.2Cl.sub.2:
C, 47.32; H, 5.56; N, 12.43. Found: C, 47.19; H, 4.78; N,
12.09.
22.2:
4-amino-7-(3'-acetyl-cis-5'-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-di-
oxaphosphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]p-
yrimidine
[0950] ##STR156##
[0951] R.sub.f=0.35 (10% MeOH in CH.sub.2Cl.sub.2). mp
90-93.degree. C. Anal calcd for
C.sub.23H.sub.26N.sub.4O.sub.8ClP.1.0.H.sub.2O: C, 48.39; H, 4.94;
N, 9.81. Found: C, 48.79; H, 4.85; N, 9.91.
Example 23
General Procedure for Preparation of 2',3'-cyclic Carbonate
Prodrugs of 2'-C-beta-methyl-7-deazaadenosine Cyclic Prodrugs
[0952] To a solution of 5'-substituted cyclic propyl prodrug (0.25
mmol) in DMF (2.5 mL) was added carbonyl diimidazole (CDI) (0.5
mmol) at 0.degree. C. The reaction was warmed to room temperature
and stirred for 4 h. Solvent was removed under reduced pressure and
the crude product was chromatographed to give 2',3'-carbonate as a
solid.
23.1: 4-amino-7-(2',
3'-carbonyl-cis-5'-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorina-
n-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
[0953] ##STR157##
[0954] R.sub.f=0.45 (10% MeOH in CH.sub.2Cl.sub.2). mp
127-130.degree. C. Anal calcd for
C.sub.22H.sub.22N.sub.4O.sub.8PCl.1.0 H.sub.2O: C, 47.62; H, 4.36;
N, 10.10. Found: C, 47.94; H, 4.10; N, 10.13.
23.2:
4-amino-7-(2',3'-carbonyl-cis-5'-O-[4-(S)-(pyridin-4-yl)-2-oxo-1,3,2-
-dioxaphosphorinan-2-yl]-2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3--
d]pyrimidine
[0955] ##STR158##
[0956] R.sub.f=0.4 (20% MeOH in CH.sub.2Cl.sub.2). mp
192-195.degree. C. Anal calcd for
C.sub.21H.sub.22N.sub.5O.sub.8P.1.0 H.sub.2O: C, 48.37; H, 4.64; N,
13.43. Found: C, 48.41; H, 4.39; N, 13.60.
Example 24
Preparation of 3'-L-valinyl Ester Prodrugs of
2'-C-beta-methyl-7-deazaadenosine Cyclic Prodrugs
24.1:
4-amino-7-(cis-5'-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphospho-
rinan-2-yl]-2'-C-methyl-3'-L-valinyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3--
d]pyrimidine
[0957] 5'-substituted cyclic prodrug (16.5) was made as described
in example 16.
Step A:
[0958] To a solution of BOC-L-Val (217 mg, 1.0 mmol) in THF (5 mL)
was added carbonyl diimidazole (CDI) (162 mg, 1 mmol). The reaction
was warmed to 50.degree. C. and allowed to stir for 1 h. The
resulting mixture was added to a solution of 5'-substituted cyclic
prodrug (16.5) (0.50 mmol) in DMF (3 mL) followed by triethylamine
(1.5 mL) and 4-dimethylamino pyridine (6 mg, 0.05 mmol). The
reaction was heated at 80.degree. C. for 3 h. The reaction mixture
was concentrated under reduced pressure and the crude was extracted
with 10% MeOH--CH.sub.2Cl.sub.2. The organic extract was washed
with water, dried and concentrated. The crude residue was
chromatographed by eluting with 5%-10% MeOH--CH.sub.2Cl.sub.2 to
give 3'-BOC-L-Val adduct of 5'-cyclic propyl prodrug (200 mg).
Step B:
[0959] The BOC protected prodrug (200 mg) was dissolved in
pre-cooled 70% aqueous trifluoroacetic acid (10 mL) at 0.degree. C.
The reaction was stirred at 0.degree. C. for 3 h. The mixture was
concentrated under reduced pressure and azeotroped with ethanol
(2.times.5 mL). The crude residue was chromatographed by eluting
with 5%-20% MeOH in CH.sub.2Cl.sub.2 to give the BOC deprotected
prodrug (140 mg). ##STR159##
[0960] R.sub.f=0.35 (15% MeOH in CH.sub.2Cl.sub.2). mp
132-135.degree. C.
[0961] Anal calcd for C.sub.26H.sub.33N.sub.5O.sub.8ClP.2.3
CF.sub.3CO.sub.2H.2.1 H.sub.2O: C, 40.38; H, 4.37; N, 7.70. Found:
C, 39.94; H, 3.93; N, 7.48.
Example 25
Preparation of 6-azido Prodrug of 2'-C-beta-methyl-7-deazaadenosine
5'-monophosphate Cyclic Prodrugs
[0962]
4-Chloro-7-(2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyr-
imidine was prepared as described in WO 02/057287.
Step A:
[0963] To a solution of
4-chloro-7-(2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
(162 mg, 0.54 mmol) in DMF (5 mL) was added sodium azide (70 mg,
1.08 mmol) at room temperature. The reaction was heated to
60.degree. C. and stirred for 18 h. The mixture was concentrated
and chromatographed by eluting with CH.sub.2Cl.sub.2 to 5%
MeOH--CH.sub.2Cl.sub.2 to give the azido substitution product (102
mg).
25.1:
4-azido-7-(2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimi-
dine
[0964] ##STR160##
[0965] R.sub.f=0.4 (5% MeOH in CH.sub.2Cl.sub.2). mp
179-180.degree. C. Anal calcd for C.sub.12H.sub.14N.sub.6O.sub.4:
C, 47.06; H, 4.61; N, 27.44. Found: C, 46.97; H, 4.71; N,
27.28.
Step B:
[0966] 5'-substituted monophosphate cyclic prodrug of
4-Azido-7-(2'-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine
is made as described in Example 16.
Biological Examples
[0967] Examples of use of the method of the invention include the
following. It will be understood that these examples are exemplary
and that the method of the invention is not limited solely to these
examples.
[0968] For the purposes of clarity and brevity, chemical compounds
are referred to as synthetic example numbers in the biological
examples below.
Example A
In Vitro Activation of Prodrug Analogues by Rat Liver Microsomes.
Quantfication by By-Product Capture
[0969] The prodrug analogues were tested for activation in rat
liver microsomes by means of a prodrug byproduct capture assay.
Methods:
[0970] Prodrugs were tested for activation by liver microsomes
isolated from rats induced with dexamethasone to enhance CYP3A4
activity (Human Biologics Inc., Phoenix Ariz.). The study was
performed at 2 mg/mL rat liver microsomes, 100 mM KH.sub.2PO.sub.4,
10 mM glutathione, 25 .mu.M or 250 .mu.M compound, and 2 mM NADPH
for 0-7.5 min. in an Eppendorf Thermomixer 5436 at 37.degree. C.,
speed 6. The reactions were initiated by addition of NADPH
following a 2-min. preincubation. Reactions were quenched with 60%
methanol at 0, 2.5, 5, and 7.5 min.
L-Glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycine, a
glutathione adduct of the by-product of prodrug activation,
3-chlorophenyl vinyl ketone, was quantified following extraction of
the reaction with 1.5 volumes of methanol. The extracted samples
were centrifuged at 14,000 rpm in an Eppendorf microfuge and the
supernatant analyzed by HPLC for
L-glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycine
content. Spiked
L-glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycine
standards (1-30 .mu.M) were prepared in 2 mg/mL microsomes under
reaction conditions and then quenched and processed in an identical
fashion to unknown samples. For HPLC analysis, the loading mobile
phase buffer (Buffer A) consisted of a 9:1 ratio (v/v) of 20 mM
potassium phosphate, pH 6.2 and acetonitrile. Extract (100 .mu.L)
was injected onto a Beckman Ultrasphere ODS column (4.6.times.250
mM, part #235329). The column was eluted with a gradient to 60%
acetonitrile. The elution of
L-glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycine
(retention time 10.4 min.) was monitored at 245 nm.
Results:
Activation of Compounds in Rat Liver Microsomes
[0971] TABLE-US-00003 Activation (250 .mu.M) Compound (nmol/mg/min)
18 4.7 16.5 0.24 17.2 0.397
Conclusion:
[0972] Formation of product,
L-glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl) cysteinylglycine
indicated activation of Compound 18 prodrug at a rate of 4.7
nmol/mg/min.
Example B
In Vitro Activation of Prodrug Analogues by Rat Liver Microsomes.
Quantification by LC-MS/MS
[0973] Prodrug analogues were tested for activation to NMP in
reactions catalyzed by the microsomal fraction of rat liver.
Methods:
[0974] Prodrugs were tested for activation by liver microsomes
isolated from rats induced with dexamethasone to enhance CYP3A4
activity (Human Biologics Inc., Phoenix Ariz.). Reactions were
conducted in 0.1 M KH.sub.2PO.sub.4, pH 7.4, in the presence of 2
mM NADPH and liver microsomes (1 mg/mL). Reaction mixtures were
incubated for 5 min. in an Eppendorf Thermomixer 5436 (37.degree.
C., speed 6). Reactions were terminated by the addition of 1.5
volumes of methanol. The resulting extracts were clarified by
centrifugation at 14,000 rpm in an Eppendorf microfuge (20 min.).
The supernatants (200 .mu.L) were evaporated under vacuum and heat
to dryness. The dried residue was reconstituted with 200 .mu.L of
water and the mixture was centrifuged for 10 min at 14,000 rpm. A
mixture of 35 .mu.L aliquot of supernatant and 35 .mu.L of mobile
phase A (20 mM N--N-dimethylhexylamine and 10 mM propionic acid in
20% methanol) was analyzed by LC-MS/MS (Applied Biosystems, API
4000) equipped with an Agilent 1100 binary pump and a LEAP
injector. NMP was detected by using MS/MS mode (M.sup.-/78.8) and
quantified based on comparison to a standard of lamivudine
monophosphate.
Results:
Activation of Compounds in Rat Liver Microsomes
[0975] TABLE-US-00004 Activation (250 .mu.M) Compound (nmol/mg/min)
16.2 0.158 16.3 0.159 16.4 0.020 16.5 0.195 16.7 0.365 17.2 1.764
16.8 0.160 16.9 0.126 16.10 0.077 16.11 0.142 16.12 0.070 16.13
0.001 16.14 0.082 16.15 0.215 16.16 0.070 16.17 0.006 16.18 0.058
16.19 0.213 16.20 0.063 16.21 0.040 16.22 0.081 16.23 0.001 16.24
0.004 16.25 0.068 16.26 0.256 16.27 0.286 16.28 0.121 17.3 1.119
16.29 0.172 17.4 0.862 17.5 1.173 17.6 1.758 16.30 0.108 16.31
0.217 16.32 0.186 17.7 0.761 17.8 0.264 17.9 0.488 17.10 1.033
17.11 1.996 17.12 0.918 17.13 1.039 17.14 1.636 17.15 0.969 17.16
0.863 17.18 0.095 17.20 1.091 17.21 0.623 17.22 0.599 17.23 0.094
17.24 0.240
Example C
In Vitro Activation in Human Liver Microsomes. Quantification by
By-Product Capture
[0976] The prodrug analogues are tested for activation in human
liver microsomes.
Methods:
[0977] Human liver microsomes are purchased from In Vitro
Technologies (IVT1032). The study is performed at 2 mg/mL human
liver microsomes, 100 mM KH.sub.2PO.sub.4, 10 mM glutathione, 25
.mu.M or 250 .mu.M compound, and 2 mM NADPH for 0-7.5 min. in an
Eppendorf Thermomixer 5436 at 37.degree. C., speed 6. The reactions
are initiated by addition of NADPH following a 2-min.
preincubation. Reactions are quenched with 60% methanol at 0, 2.5,
5, and 7.5 min.
L-Glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycine, a
glutathione adduct of the by-product of prodrug activation,
3-chlorophenyl vinyl ketone, is quantified following extraction of
the reaction with 1.5 volumes of methanol. The extracted samples
are centrifuged at 14,000 rpm in an Eppendorf microfuge and the
supernatant analyzed by HPLC for
L-glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycine
content. Spiked
L-glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycine
standards (1-30 .mu.M) are prepared in 2 mg/mL microsomes under
reaction conditions and then quenched and processed in an identical
fashion to unknown samples. For HPLC analysis, the loading mobile
phase buffer (Buffer A) consists of a 9:1 ratio (v/v) of 20 mM
potassium phosphate, pH 6.2 and acetonitrile. Extract (100 .mu.L)
is injected onto a Beckman Ultrasphere ODS column (4.6.times.250
mM, part #235329). The column is eluted with a gradient to 60%
acetonitrile. The elution of
L-glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycine
(retention time 10.4 min.) is monitored at 245 nm.
Conclusion:
[0978] Formation of product,
L-glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl) cysteinylglycine
indicates the prodrugs are activated in vitro in human liver
microsomes.
Example D
In Vitro Activation of Prodrug Analogues by Human Liver Microsomes.
Quantification by LC-MS/MS
[0979] Prodrug analogues were tested for activation to NMP in
reactions catalyzed by the microsomal fraction of human liver.
Methods:
[0980] Prodrugs were tested for activation by human liver
microsomes purchased from In Vitro Technologies (IVT1032) Reactions
were conducted in 0.1 M KH.sub.2PO.sub.4, pH 7.4, in the presence
of 2 mM NADPH and liver microsomes (1 mg/mL). Reaction mixtures
were incubated for 5 min. in an Eppendorf Thermomixer 5436
(37.degree. C., speed 6). Reactions were terminated by the addition
of 1.5 volumes of methanol. The resulting extracts were clarified
by centrifugation at 14,000 rpm in an Eppendorf microfuge (20
min.). The supernatants (200 .mu.L) were evaporated under vacuum
and heated to dryness. The dried residue was reconstituted with 200
.mu.L of water and the mixture was centrifuged for 10 min at 14,000
rpm.
[0981] A mixture of 35 .mu.L aliquot of supernatant and 35 .mu.L of
mobile phase A (20 mM N--N-dimethylhexylamine and 10 mM propionic
acid in 20% methanol) was analyzed by LC-MS/MS (Applied Biosystems,
API 4000) equipped with an Agilent 1100 binary pump and a LEAP
injector. NMP was detected by using MS/MS mode (M.sup.-/78.8) and
quantified based on comparison to a standard of lamivudine
monophosphate.
Results:
Activation of Compounds in Human Liver Microsomes
[0982] TABLE-US-00005 Activation (250 .mu.M) Compound (nmol/mg/min)
16.2 0.301 16.3 0.162 16.4 0.049 16.5 0.463 16.7 0.213 17.2 2.040
16.8 0.436 16.9 0.316 16.10 0.241 16.11 0.100 16.12 0.394 16.13
0.002 16.14 0.282 16.15 0.335 16.16 0.075 16.17 0.021 16.18 0.044
16.19 0.171 16.20 0.137 16.21 0.043 16.22 0.077 16.23 0.013 16.24
0.031 16.25 0.242 16.26 0.223 16.27 0.455 16.28 0.293 17.3 1.677
17.4 1.324 17.5 0.952 17.6 2.086 16.30 0.037 16.31 0.138 16.32
0.074 17.7 1.024 17.8 0.322 17.9 0.314 17.10 0.626 17.11 1.439
17.12 0.750 17.13 0.499 17.14 1.164 17.15 0.733 17.16 0.497 17.18
0.085 17.20 1.381 17.21 0.626 17.22 0.484 17.23 0.089 17.24
0.455
Example E
NTP Accumulation in Hepatocytes Following Incubation with
Nucleoside Analogues and their Prodrugs
[0983] Nucleoside analogues and their prodrugs were evaluated for
their ability to generate NTPs in freshly isolated rat hepatocytes.
It is generally accepted that the NTP form of a nucleoside is the
active antiviral agent.
Methods:
[0984] Hepatocytes were prepared from fed Sprague-Dawley rats
(250-300 g) according to the procedure of Berry and Friend (Berry,
M. N. Friend, D. S., J. Cell Biol. 43:506-520 (1969)) as modified
by Groen (Groen, A. K. et al., Eur. J. Biochem 122:87-93 (1982)).
Hepatocytes (20 mg/mL wet weight, >85% trypan blue viability)
were incubated at 37.degree. C. in 2 mL of Krebs-bicarbonate buffer
containing 20 mM glucose, and 1 mg/mL BSA for 2 h in the presence
of 1-250 .mu.M nucleoside or prodrug (from 25 mM stock solutions in
DMSO). Following the incubation, 1600 .mu.L aliquot of the cell
suspension was centrifuged and 300 .mu.L of acetonitrile was added
to the pellet, vortexed and sonicated until the pellet broke down.
Then 200 .mu.L of water was added to make a 60% acetonitrile
solution. After 10 min centrifugation at 14,000 rpm, the resulting
supernatant was transferred to a new vial and evaporated to near
dryness in a Savant SpeedVac Plus at room temperature. The dried
residue was reconstituted with 200 .mu.L of water and the mixture
was centrifuged for 10 min at 14,000 rpm. A mixture of 35 .mu.L
aliquot of supernatant and 35 .mu.L of mobile phase A (20 mM
N--N-dimethylhexylamine and 10 mM propionic acid in 20% methanol)
was analyzed by LC-MS/MS (Applied Biosystems, API 4000) equipped
with an Agilent 1100 binary pump and a LEAP injector. NTP was
detected by using MS/MS mode (M.sup.-/78.8) and quantified based on
comparison to a standard of lamivudine triphosphate.
Results:
[0985] Following the incubation of 25 .mu.M or 250 .mu.M
nucleosides and prodrugs with primary rat hepatocytes, NTP
formation observed over the course of 2 h was measured as nmol/g.
TABLE-US-00006 NTP formation from NTP formation from 25 .mu.M
compound 250 .mu.M compound Compound (nmol/g) (nmol/g) 2'-C- 193
798 methyladenosine 2'-C- 1.3 4.7 methylguanosine 19 13.8 56.7 18
51.8 16.1 85 160 19 130.9 16.2 130.3 16.3 102.6 16.4 55.0 16.7
347.7 17.2 4.2 16.8 160.6 16.9 115.1 16.10 17.6 16.11 63.2 16.12
18.3 16.13 3.4 16.14 18.6 16.15 34.5 16.16 6.3 16.17 7.2 16.18 13.8
16.19 70.1 16.20 19.4 16.21 10.2 16.22 10.9 16.23 2.7 16.24 4.4
16.25 22.5 16.26 58.5 16.27 63.9 16.28 19.8 17.3 2.2 16.29 14.1
17.4 3.1 17.5 2.0 17.6 3.5 16.30 36.7 16.31 16.5 16.32 49.5 17.7
1.3 17.8 1.6 17.9 2.0 17.10 2.1 17.11 3.4 17.12 1.9 17.13 3.8 17.14
4.0 17.15 1.8 17.16 1.5 17.17 0.2 17.18 4.1 17.19 0.1 17.20 2.1
17.21 1.2 17.22 1.8 17.23 0.3 17.24 3.9
Conclusion:
[0986] Compounds of this invention showed an ability to generate
NTP in freshly isolated rat hepatocytes.
Example F
HCV-Infected Human Liver Slice Assay
[0987] Inhibition of HCV replication in human liver tissue was
evaluated using the following assay.
Methods:
Procurement:
[0988] Liver from a brain-dead HCV antibody-positive human patient
was perfused with ice-cold Viaspan (Dupont Pharmaceutical)
preservation solution and received on ice in Viaspan.
[0989] Precision-cut liver slices of .about.200-250 .mu.m thickness
and 8 cm diameter were prepared and cultured in Waymouth's cell
culture medium (Gibco, Inc.) that was supplemented with 10% fetal
bovine serum and 10 mL/L Fungi-Bact at 37.degree. C., and gassed
with carbogen (95% O.sub.2, 5% CO.sub.2) at 0.75 liters/min. Tissue
slices were maintained in culture for 72 h. Cell culture medium
containing test compound in solution was changed on a daily
basis.
[0990] At appropriate times of liver slice incubation, liver slices
and medium were collected for analysis of HCV RNA (tissue and
medium) and nucleotide levels (NTP). All collected media and tissue
slices were maintained in liquid N.sub.2 until analysis.
[0991] Medium and tissue samples were analyzed for HCV RNA levels
according to published procedures (Bonacini et. al., 1999) which
utilize an automated, multicycle, polymerase chain reaction
(PCR)-based technique. This assay has a lower limit of detection
for HCV RNA of 100 viral copies/ml.
Analysis of Tissue NTPs:
[0992] Frozen liver slices were disrupted by using a combination of
ultrasound probe sonication, Branson Sonifier 450 (Branson
Ultrasonics, Danbury, Conn.) and homogenization using a Dounce
conical pestle in 200 .mu.ls of 10% (v/v) perchloric acid (PCA).
After a 5 min centrifugation at 2,500.times.g, the supernatants
were neutralized using 3 M KOH/3 M KHCO.sub.3 and mixed thoroughly.
The neutralized samples were centrifuged at 2,500 g for 5 min and
NTP levels were determined by ion exchange phase HPLC (Hewlett
Packard 1050) using a Whatman Partisil 5 SAX (5 .mu.m,
4.6.times.250 mm) column. Samples (50 .mu.L) were injected onto the
column in 70% 10 mM ammonium phosphate buffer and 30% 1 M ammonium
phosphate buffer, both at pH 3.5 and containing 6% ethanol.
Nucleoside triphosphates were eluted from the column using a linear
gradient to 80% 1 M ammonium phosphate pH 3.5/6% ethanol buffer, at
a flow rate of 1.25 mL/min and detected by UV absorbance (254
nm).
Results:
[0993] HCV RNA levels present in the liver slice culture media
decreased from the levels present in control, untreated slices
following incubation with 2'-C-methylguanosine and compound 19.
TABLE-US-00007 Log.sub.10 decrease Log.sub.10 decrease in viral RNA
in viral RNA Concentration of from control at 48-72 h from control
at 48-72 h Compound following treatment with 2'- following
treatment with (.mu.M) C-methylguanosine compound 19 0.25 0.51 1.27
1 -- 1.61 2.5 1.74 1.70 25 1.48 1.72
Conclusion:
[0994] Treatment of HCV-infected human liver slices with
2'-C-methylguanosine or compound 19 for 72 h decreased the amount
of HCV RNA released into the culture medium from 48-72 h. Treatment
with the prodrug, compound 19 was more effective than treatment
with the nucleoside, 2'-C-methylguanosine, at lowering viral RNA
production in the culture medium.
Example G
Liver Targeting of Nucleoside Analogues and their Prodrugs
[0995] The liver specificity of the compound 19 prodrug was
compared relative to the parent nucleoside, 2'-C-methylguanosine,
and for compound 21.1 prodrug relative to its parent nucleoside,
2'-C-methyladenosine, by measuring the generation of NTP in the
liver compared to nucleoside in the plasma.
Methods:
[0996] Compound 19 or 2'-C-methylguanosine were administered
intraperitoneally to C57BL/6 mice at a dose of 30 mg/kg based on
nucleoside equivalents (30 mg/kg 2'-C-methylguanosine and 53.27
mg/kg compound 19). Compound 21.1 or 2'-C-methyladenosine were
administered intravenously to C57BL/6 mice at dose of about 5.5
mg/kg nucleoside equivalents (5.5 mg/kg 2'-C-methyladenosine and 10
mg/kg compound 21.1). Plasma concentrations of
2'-C-methylguanosine, compound 19, 2'-C-methyladenosine, and
compound 21.1 were determined by HPLC-UV and the liver
concentrations of the 5'-triphosphate of 2'-C-methylguanosine and
2'-C-methyladenosine were measured by LC-MS using the standard
ion-pairing chromatography method for triphosphate as described in
Example E. Conventional SAX HPLC-UV could not differentiate between
endogenous GTP and 2'-C-methylguanosine triphosphate. Since an
authentic standard of 2'-C-methylguanosine triphosphate was not
available, the liver concentrations of the nucleotide were
approximated as described in Example E.
Results:
[0997] Liver targeting of compound 19 as the triphosphate of
2'-C-methylguanosine and of compound 21.1 as the triphosphate of
2'-C-methyladenosine were clearly demonstrated with the prodrugs.
The relative liver NTP AUC values, plasma nucleoside AUC values,
the liver targeting ratio (liver/plasma), and the fold-improvement
with the prodrugs are summarized in the table below. Compound 19
showed a 30-fold prodrug improvement of liver targeting over free
nucleoside. Compound 21.1 showed a greater than 32-fold prodrug
improvement of liver targeting over free nucleoside, which was
below the limit of quantitation in the plasma following dosing of
compound 21.1. TABLE-US-00008 Liver Plasma Targeting Liver
Nucleoside Index Prodrug Nucleoside NTP AUC AUC (Liver/ Improvement
[Prodrug] (nmol * h/g) (.mu.M * h) Plasma) (Fold) 2'-C- 64 73.7
0.87 -- methylguanosine 19 485 18.5 26.2 30 2'-C- 119 26.8 4.4 --
methyladenosine 21.1 502 <3.6 >141.4 >32
Example H
Tissue Distribution Following Oral Administration of Nucleoside
Analogues and their Prodrugs
[0998] The liver specificity of prodrugs are compared relative to
their parent nucleoside analog inhibitors in liver and other organs
that could be targets of toxicity.
Methods:
[0999] Nucleoside analogues and their prodrugs are administered at
30 mg/kg (in terms of nucleoside equivalents) to fasted rats by
oral gavage. Plasma concentrations of nucleoside and prodrug are
determined by HPLC-UV, as described in Example J, and the liver,
skeletal muscle, cardiac, kidney, small intestine, and other organ
concentrations of the 5'-triphosphate of the nucleoside are
measured by LC-MS using the standard ion-pairing chromatography
method for triphosphate as described in Example E.
Results:
[1000] The results demonstrate the liver targeting of the
nucleoside analog prodrugs and provide evidence for improved safety
of the prodrugs over that of the nucleosides alone. This can occur
solely by the liver targeting provided by the prodrug or by
additional liver metabolism of nucleoside derived following
dephosphorylation of the nucleoside monophosphate. In the latter
case, the nucleoside can escape from the liver into the periphery
leading to exposure of other tissues to the nucleoside and
potential extrahepatic toxicity. The release of nucleoside from the
liver can be reduced by metabolism of the nucleoside monophosphate
or the nucleoside in the liver cell, e.g. the breakdown of
adenosine-based nucleoside monophosphates to inosine via adenylate
deaminase and nucleotidase, or the breakdown of adenosine-based
nucleoside to inosine and hypoxanthine by adenosine deaminase and
purine nucleoside phosphorylase, respectively.
Example I
Assessment of the Oral Bioavailability of Nucleoside Analogues and
their Prodrugs in Normal Male Rats
[1001] The oral bioavailability (OBAV) of the nucleoside analogues
and their prodrugs were evaluated in the normal male rat.
Methods:
[1002] The compounds were solubilized in a suitable vehicle for
intravenous and oral administration. The OBAV was assessed by
calculating the ratio of the AUC values of the liver organ
concentration-time profile of NTP following oral and i.v. or i.p.
administration of 30 mg/kg (in terms of nucleoside equivalents) of
the compound to groups of four rats. Liver organ samples were taken
at 20 min and 1, 3, 5, 8, 12, and 24 h following dosing. Liver
organ concentrations of NTP were determined as determined by
LC-MS/MS (Example E) or HPLC (Example F) analysis.
Results:
Oral Bioavailability in the Normal Male Rat
[1003] TABLE-US-00009 Compound % F 16.32 31.7
Example J
Susceptibility of Nucleoside Analogues to Metabolism in Rat Liver
S9Fraction or Isolated Hepatocytes
[1004] The susceptibility of the purine nucleoside analogues to
metabolism is assessed in rat liver S9 fraction or isolated rat
hepatocytes.
[1005] Methods: Purine nucleoside analogues (100 .mu.M) (e.g.,
2'-C-methyladenosine) are incubated in rat liver S9 fraction or
with isolated rat hepatocytes at 37.degree. C. The reactions are
terminated at time points up to 2 h and then deproteinized by
extraction with 60% acetonitrile. Following centrifugation, the
supernatants are evaporated to dryness and the resulting residues
are reconstituted with aqueous mobile phase. These samples are
analyzed for potential metabolites by a single HPLC system equipped
with a diode array detector. Nucleosides (e.g., 2'-C-methylinosine)
and bases (e.g., hypoxanthine) are separated and quantified on a
Beckman Ultrasphere C-18 reverse phase column (4.5.times.250 mm)
using a gradient of Buffer A (100 mM potassium phosphate pH 6) and
Buffer B (25% v/v methanol) at a flow rate of 1.5 mL/min. The
column is developed over 40 min using a nonlinear gradient of 0%
Buffer B to 100% Buffer B (% pump Buffer B=100.times.(time
[min]/40).sup.3) and monitored by UV absorbance at 260 nm.
Metabolites are identified by coelution with authentic standards
and/or UV spectrum matching.
[1006] Results: The susceptibility of the purine nucleoside
analogues to metabolism is dependent upon the type and location of
the structural modification of the congener. The inclusion of
certain pharmacophores (such as the 2'-C-methyl group of
2'-C-methyladenosine] leads to increased resistance to metabolism
by purine salvage pathway enzymes [such as adenosine deaminase and
purine nucleoside phosphorylase)..sup.1 .sup.1Eldrup A B, Allerson
C R, Bennett C F, et al. (2004) J. Med. Chem. 47(9):2283-2295,
"Structure-activity relationship of purine ribonucleotides for
inhibition of hepatitis C virus RNA-dependent RNA polymerase."
* * * * *