U.S. patent application number 11/908019 was filed with the patent office on 2010-11-18 for bicyclic nucleosides and nucleotides as therapeutic agents.
This patent application is currently assigned to Biota Scientific Management Pty Ltd. Invention is credited to Silas Bond, Alistair Draffan, Paula Francom, John Lambert, Roland Nearn.
Application Number | 20100291031 11/908019 |
Document ID | / |
Family ID | 36952869 |
Filed Date | 2010-11-18 |
United States Patent
Application |
20100291031 |
Kind Code |
A2 |
Francom; Paula ; et
al. |
November 18, 2010 |
BICYCLIC NUCLEOSIDES AND NUCLEOTIDES AS THERAPEUTIC AGENTS
Abstract
The invention relates to the use of bicyclic nucleosides and
nucleotides based on formula (II) for the treatment of infectious
diseases, and in particular, viral infections.
Inventors: |
Francom; Paula; (Notting
Hill, Victoria, AU) ; Nearn; Roland; (Chelsea
Heights, Victoria, AU) ; Draffan; Alistair; (St Kilda
East, Victoria, AU) ; Lambert; John; (Blackburn
South, Victoria, AU) ; Bond; Silas; (Lynbrook,
Victoria, AU) |
Correspondence
Address: |
SEED INTELLECTUAL PROPERTY LAW GROUP PLLC
701 FIFTH AVE
SUITE 5400
SEATTLE
WA
98104
UNITED STATES
206-622-4900
206-682-6031
|
Assignee: |
Biota Scientific Management Pty
Ltd
585 Blackburn Road, Unit 10
Notting Hill, Victoria
AU
3168
|
Prior
Publication: |
|
Document Identifier |
Publication Date |
|
US 20090004138 A1 |
January 1, 2009 |
|
|
Family ID: |
36952869 |
Appl. No.: |
11/908019 |
Filed: |
March 8, 2006 |
PCT Filed: |
March 8, 2006 |
PCT NO: |
PCT/AU2006/000303 |
371 Date: |
April 9, 2008 |
Related U.S. Patent Documents
|
|
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|
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Application
Number |
Filing Date |
Patent Number |
|
|
60/661,665 |
Mar 8, 2005 |
|
|
|
Current U.S.
Class: |
424/85.4 ;
514/43; 536/22.1; 536/27.13 |
Current CPC
Class: |
A61P 31/06 20180101;
A61P 31/14 20180101; C07H 19/22 20130101; C07H 19/23 20130101; A61K
31/706 20130101; A61P 31/04 20180101 |
Class at
Publication: |
424/085.4 ;
536/027.13; 536/022.1; 514/043 |
International
Class: |
A61K 31/7068 20060101
A61K031/7068; C07H 19/23 20060101 C07H019/23; A61P 31/14 20060101
A61P031/14; A61K 38/21 20060101 A61K038/21 |
Claims
1. A method for the treatment of a microbial infection comprising
administering an effective amount of a compound of the formula (I)
which may be a D- or L-nucleoside or nucleotide, or a
pharmaceutically acceptable salt thereof, ##STR51## wherein: A is
O, S, CH.sub.2, CHF, CF.sub.2 or NR; R.sup.1, R.sup.2, R.sup.2',
R.sup.3, R.sup.3', and R.sup.4 are independently selected from the
group consisting of H, halogen, OH, N.sub.3, CN, optionally
substituted alkyl, optionally substituted alkenyl, optionally
substituted alkynyl, optionally substituted aryl, optionally
substituted acyl, optionally substituted arylalkyl, optionally
substituted heterocyclyl, optionally substituted heteroaryl,
optionally substituted cycloalkyl, optionally substituted
cycloalkenyl, optionally substituted alkyloxy, optionally
substituted alkenyloxy, optionally substituted alkynoxy, optionally
substituted aryloxy, optionally substituted acyloxy, optionally
substituted oxyacyl, optionally substituted arylalkoxy, optionally
substituted heterocycloxy, optionally substituted heteroaryloxy,
optionally substituted cycloalkoxy, optionally substituted
cycloalkenoxy, optionally substituted amino, optionally substituted
aminoacyl, optionally substituted aminoacyloxy, optionally
substituted acylamino, optionally substituted oxyacylamino,
optionally substituted oxyacyloxy, optionally substituted
acylimino, optionally substituted acyliminoxy, optionally
substituted oxyacylimino, optionally substituted aminothioacyl,
optionally substituted thioacylamino, optionally substituted
aminosulfinyl, optionally substituted aminosulfonyl, optionally
substituted thio, optionally substituted thioacyl, optionally
substituted thioacyloxy, optionally substituted oxythioacyl,
optionally substituted oxythioacyloxy, optionally substituted
phosphorylamino, optionally substituted sulfinyl, optionally
substituted sulfonyl, optionally substituted sulfinylamino,
optionally substituted sulfonylamino, optionally substituted
oxysulfinylamino, and optionally substituted oxysulfonylamino, or
R.sup.2 and R.sup.2' together or R.sup.3 and R.sup.3' together
represents .dbd.O, .dbd.S, or =L-Y' where L is N, CH, CF, CCl or
CBr and Y' is H, halogen, N.sub.3, methyl, ethyl or CN; R.sup.4' is
--CY.sub.2SH, --CY.sub.2OH, --CY.sub.2NH.sub.2, or -L-R.sup.5; L'
is selected from the group consisting of --CY.sub.2--,
--CY.sub.2CY.sub.2--, --CY.sub.2OCY.sub.2--, --CY.sub.2SCY.sub.2--
and --CY.sub.2NHCY.sub.2--; each Y is independently selected from
the group consisting of H, F, Cl, Br, OR, C.sub.1-C.sub.6alkyl,
C.sub.2-C.sub.6alkenyl and C.sub.2-C.sub.6alkynyl, wherein
C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl, and
C.sub.2-C.sub.6alkynyl may be optionally substituted with one or
more groups selected from F, Cl, Br, OH, COOH, COOCH.sub.3, SH,
SCH.sub.3, NH.sub.2, NHCH.sub.3, N(CH.sub.3).sub.2, CN, NO.sub.2,
C(O)NH.sub.2, C(O)NHCH.sub.3, N.sub.3, C(S)NH.sub.2, OCH.sub.3, and
OCH.sub.2CH.sub.3; R.sup.5 is selected from the group consisting of
OR, NR.sub.2, monophosphate, diphosphate, and triphosphate, or a
mono, di or triphosphate mimic; each R is independently selected
from the group consisting of H, CF.sub.3, optionally substituted
alkyl, optionally substituted alkenyl, optionally substituted
alkynyl, optionally substituted aryl, optionally substituted acyl,
optionally substituted cycloalkyl, optionally substituted
cycloalkenyl, optionally substituted heteroaryl, optionally
substituted heterocyclyl, and optionally substituted arylalkyl; B
is selected from a group of formula (I) ##STR52## wherein, if Z is
a participant in a .pi. bond (double bond), Z is independently
selected from N or C-G; or, if Z is not a participant in a .pi.
bond (double bond), Z is independently selected from O, S, Se, NR,
NOR, NNR.sub.2, CO, CS, CNR, SO, S(O).sub.2, SeO, Se(O).sub.2 or
C(G).sub.2; each G is independently selected from the group
consisting of H, halogen, OR, SR, NR.sub.2, NROR, N.sub.3, COOR,
CN, CONR.sub.2, C(S)NR.sub.2, C(.dbd.NR)NR.sub.2, and R; and where
any two adjacent Z are not both selected from O, S, and Se, or not
both selected from CO, CS, CNR, SO, S(O).sub.2, SeO, and
Se(O).sub.2; wherein, if X is a participant in a .pi. bond (double
bond), X is C; or if X is not a participant in a .pi. bond (double
bond), X is CR or N; wherein, if R'' is a participant in a .pi.
bond (double bond), R'' is O, S, Se, NR, NOR or NNR.sub.2; or if
R'' is not a participant in a .pi. bond (double bond), R'' is OR,
SR, F, Cl, R, or SeR; and dashed lines (---) indicate a possible
.pi. or double bond; optionally in combination with one or more
antiviral, antibacterial, or antiproliferative agents.
2. A method according to claim 1 for the treatment of a microbial
infection which is a viral infection.
3. A method according to claim 2 wherein the viral infection is
caused by an RNA virus.
4. A method according to claim 3 wherein the viral infection is
caused by an RNA virus of the group Flaviviridae.
5. A method according to claim 4 wherein the virus is the hepatitis
C virus.
6. A method according to claim 1, wherein the compound of formula
(I) has a base structure B selected form the following formulae
(XI) to (XXI) ##STR53## ##STR54## wherein Z, X and R'' are as
defined in claim 1.
7. A method according to claim 6, wherein X is N.
8. A method according to claim 1, wherein the compound of formula
(I) has a base structure B selected from the following: ##STR55##
##STR56## ##STR57## wherein G and R are as defined in claim 1.
9. A method according to claim 1, wherein the compound of formula
(I) has a base structure B represented by formula IIa: ##STR58##
wherein each Z' is independently N (if a participant in a .pi.
bond) or NR (if not a participant in a .pi. bond), and R'', R and Z
are as defined in claim 1.
10. A method according to claim 1, wherein the compound of formula
(I) has a base structure B represented by the formula IIb:
##STR59## wherein each Z' is independently N (if a participant in a
.pi. bond) or NR (if not a participant in a .pi. bond), and each Z
is independently CG (if a participant in a .pi. bond) or
>C(G).sub.2 (if not a participant in a .pi. bond), wherein R''
and G are as defined in claim 1.
11. A method according to claim 1, wherein the compound of formula
(I) has a base structure B represented by formula IIc: ##STR60##
wherein R and G are as defined in claim 1.
12. A method according to claim 1, wherein the compound of formula
(I) has a base structure B represented by the formula IId:
##STR61## wherein R and G are as defined in claim 1.
13. A method according to claim 1, wherein the compound of formula
(I) has a base structure B represented by the formula IIe:
##STR62## wherein R and G are as defined in claim 1.
14. A method according to claim 1, wherein at least one of R.sup.2
and R.sup.2' of the compound of formula (I) is methyl, hydroxyl or
F.
15. A method according to claim 1, wherein L' of the compound of
formula (I) is --CH.sub.2--.
16. A method according to claim 1, wherein R.sup.4' of the compound
of formula (I) is selected from --CH.sub.2--OH, --CF.sub.2OH,
--CCl.sub.2--OH, --C(CH.sub.3)(CH).sub.3OH, --CH(CH.sub.3)OH,
--CH.sub.2--CH.sub.2--P(O)(OH).sub.2,
--CH.sub.2--CH.sub.2--P(O)(OH).sub.2, --CH.sub.2SP(O)(OH).sub.2,
--CH.sub.2SH, --CF.sub.2SH, and
--CH.sub.2--O--P(O)(OPh)(NHCH(CH.sub.3)(CO.sub.2Me)).
17. A method according to claim 16, wherein R.sup.4' is
--CH.sub.2--OH.
18. A method according to claim 1, wherein the sugar moiety of the
compound of formula (I) is selected from the following formulae:
##STR63## or C-5 monophosphate, diphosphate and triphosphate
derivatives thereof, or C-5 mono, di or triphosphate mimics,
wherein R is as defined in claim 1.
19. A method according to claim 1, wherein the sugar moiety of the
compound of formula (I) is selected from the following formulae:
##STR64## or C-5 monophosphate, diphosphate and triphosphate
derivatives thereof, or C-5 mono, di or triphosphate mimics,
wherein R is as defined in claim 1.
20. A method according to claim 1, wherein the sugar moiety of the
compound of formula (I) is represented by the formula: ##STR65## or
C-5 monophosphate, diphosphate and triphosphate derivatives
thereof, or C-5 mono, di or triphosphate mimics, wherein R is as
defined in claim 1.
21. A method according to claim 1, wherein the sugar moiety of the
compound of formula (I) is represented by the formula: ##STR66## or
C-5 monophosphate, diphosphate and triphosphate derivatives
thereof, or C-5 mono, di or triphosphate mimics.
22. A method according to claim 1, wherein the compound of formula
(I) is selected from ##STR67## ##STR68## ##STR69## wherein: each R
on the sugar moiety is independently selected from H, optionally
substituted alkyl, optionally substituted alkenyl, optionally
substituted alkynyl, optionally substituted aryl, optionally
substituted acyl and optionally substituted arylalkyl; each R on
the base moiety is independently selected from H, optionally
substituted alkyl, optionally substituted aryl, optionally
substituted heteroaryl, optionally substituted heterocyclyl, and
optionally substituted arylalkyl; each G is independently selected
from H, halogen, CF.sub.3, optionally substituted alkyl, optionally
substituted aryl, optionally substituted heteroaryl, optionally
substituted heterocyclyl, and optionally substituted arylalkyl; and
C-5 monophosphate, diphosphate and triphosphate derivatives
thereof, or C-5 mono, di or triphosphate mimics.
23. A method according to claim 1, wherein the compound of formula
(I) is selected from ##STR70## ##STR71## ##STR72##
24. A method according to claim 1 which includes the administration
of a compound of formula (I) or a pharmaceutically acceptable salt
thereof in combination with one or more antiviral agents.
25. A method according to claim 24, wherein the antiviral agents
are selected from interferons and interferon derivatives, IMPDH
inhibitors, antiviral nucleosides, polymerase inhibitors and
protease inhibitors.
26-48. (canceled)
49. A compound of the formula (I) which may be a D- or L-nucleoside
or nucleotide, or a salt thereof; ##STR73## wherein: A is O, S,
CH.sub.2, CHF, CF.sub.2 or NR; R.sup.1, R.sup.2, R.sup.2', R.sup.3,
R.sup.3', and R.sup.4 are independently selected from the group
consisting of H, halogen, OH, N.sub.3, CN, optionally substituted
alkyl, optionally substituted alkenyl, optionally substituted
alkynyl, optionally substituted aryl, optionally substituted acyl,
optionally substituted arylalkyl, optionally substituted
heterocyclyl, optionally substituted heteroaryl, optionally
substituted cycloalkyl, optionally substituted cycloalkenyl,
optionally substituted alkyloxy, optionally substituted alkenyloxy,
optionally substituted alkynoxy, optionally substituted aryloxy,
optionally substituted acyloxy, optionally substituted oxyacyl,
optionally substituted arylalkoxy, optionally substituted
heterocycloxy, optionally substituted heteroaryloxy, optionally
substituted cycloalkoxy, optionally substituted cycloalkenoxy,
optionally substituted amino, optionally substituted aminoacyl,
optionally substituted aminoacyloxy, optionally substituted
acylamino, optionally substituted oxyacylamino, optionally
substituted oxyacyloxy, optionally substituted acylimino,
optionally substituted acyliminoxy, optionally substituted
oxyacylimino, optionally substituted aminothioacyl, optionally
substituted thioacylamino, optionally substituted aminosulfinyl,
optionally substituted aminosulfonyl, optionally substituted thio,
optionally substituted thioacyl, optionally substituted
thioacyloxy, optionally substituted oxythioacyl, optionally
substituted oxythioacyloxy, optionally substituted phosphorylamino,
optionally substituted sulfinyl, optionally substituted sulfonyl,
optionally substituted sulfinylamino, optionally substituted
sulfonylamino, optionally substituted oxysulfinylamino, and
optionally substituted oxysulfonylamino, or R.sup.2 and R.sup.2'
together or R.sup.3 and R.sup.3' together represents .dbd.O,
.dbd.S, or =L-Y' where L is N, CH, CF, CCl or CBr and Y' is H,
halogen, N.sub.3, methyl, ethyl or CN; R.sup.4' is --CY.sub.2SH,
--CY.sub.2OH, --CY.sub.2NH.sub.2, or -L-R.sup.5; L' is selected
from the group consisting of --CY.sub.2--, --CY.sub.2CY.sub.2--,
--CY.sub.2OCY.sub.2--, --CY.sub.2SCY.sub.2-- and
--CY.sub.2NHCY.sub.2--; each Y is independently selected from the
group consisting of H, F, Cl, Br, OR, C.sub.1-C.sub.6alkyl,
C.sub.2-C.sub.6alkenyl and C.sub.2-C.sub.6alkynyl, wherein
C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl, and
C.sub.2-C.sub.6alkynyl may be optionally substituted with one or
more groups selected from F, Cl, Br, OH, COOH, COOCH.sub.3, SH,
SCH.sub.3, NH.sub.2, NHCH.sub.3, N(CH.sub.3).sub.2, CN, NO.sub.2,
C(O)NH.sub.2, C(O)NHCH.sub.3, N.sub.3, C(S)NH.sub.2, OCH.sub.3, and
OCH.sub.2CH.sub.3; R.sup.5 is selected from the group consisting of
OR, NR.sub.2, monophosphate, diphosphate, and triphosphate, or a
mono, di or triphosphate mimic; each R is independently selected
from the group consisting of H, CF.sub.3, optionally substituted
alkyl, optionally substituted alkenyl, optionally substituted
alkynyl, optionally substituted aryl, optionally substituted acyl,
optionally substituted cycloalkyl, optionally substituted
cycloalkenyl, optionally substituted heteroaryl, optionally
substituted heterocyclyl, and optionally substituted arylalkyl; B
is a group of formula (II) ##STR74## wherein, if Z is a participant
in a .pi. bond (double bond), Z is independently selected from N or
C-G; or, if Z is not a participant in a .pi. bond (double bond), Z
is independently selected from O, S, Se, NR, NOR, NNR.sub.2, CO,
CS, CNR, SO, S(O).sub.2, SeO, Se(O).sub.2 or C(G).sub.2, wherein
each G is independently selected from the group consisting of H,
halogen, OR, SR, NR.sub.2, NROR, N.sub.3, COOR, CN, CONR.sub.2,
C(S)NR.sub.2, C(.dbd.NR)NR.sub.2, and R; and where any two adjacent
Z are not both selected from O, S, and Se, or not both selected
from CO, CS, CNR, SO, S(O).sub.2, SeO, and Se(O).sub.2; wherein, if
X is a participant in a .pi. bond (double bond), X is C; or if X is
not a participant in a .pi. bond (double bond), X is CR or N;
wherein, if R'' is a participant in a .pi. bond (double bond), R''
is O, S, Se, NR, NOR, and NNR.sub.2; or if R'' is not a participant
in a .pi. bond (double bond), R'' is OR, SR, F, Cl, R, or SeR;
dashed lines (---) indicate a possible .pi. or double bond; and
wherein when R.sup.2', R.sup.3', and R.sup.5 are OH or
OC(O)CH.sub.3, L' is CH.sub.2, A is O, and R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 are H, B is not the group of formula (III),
formula (IV), formula (V), formula (VI), or formula (VII) ##STR75##
(where each X* is H or one of X* is CH.sub.3 and the other two X*
are H); when B is a group of formula (III), formula (VIII), or
formula (IX) ##STR76## R.sup.2 and R.sup.2' are not both H; when
R.sup.2, R.sup.3' and R.sup.5 are OH, L' is CH.sub.2, A is O and
R.sup.1, R.sup.2', R.sup.3 and R.sup.4 are H, B is not the group of
formula (IV); and when R.sup.2' is F, R.sup.3' is OH, R.sup.5 is
triphosphate, L' is CH.sub.2, A is O, and R.sup.1, R.sup.2, R.sup.3
and R.sup.4 are H, B is not a group of formula (X) ##STR77##
50. A compound according to claim 49, or a salt thereof, wherein
the base structure B is selected from the following formulae (XI)
to (XXI) ##STR78## ##STR79## wherein Z, X and R'' are as defined in
claim 49.
51. A compound according to claim 49, or a salt thereof, wherein X
is N.
52. A compound according to claim 49, or a salt thereof, wherein
the base structure B is selected from the following: ##STR80##
##STR81## ##STR82## wherein G and R are as defined in claim 49.
53. A compound according to claim 49, or a salt thereof, wherein
the base structure B is represented by formula IIa: ##STR83##
wherein each Z' is independently N (if a participant in a .pi.
bond) or NR (if not a participant in a .pi. bond), and R'', R and Z
are as defined in claim 49.
54. A compound according to claim 49, or a salt thereof, wherein
the base structure B is represented by the formula IIb: ##STR84##
wherein each Z' is independently N (if a participant in a .pi.
bond) or NR (if not a participant in a .pi. bond), and each Z is
independently CG (if a participant in a .pi. bond) or
>C(G).sub.2 (if not a participant in a .pi. bond), and wherein
R'' and G is as defined in claim 49.
55. A compound according to claim 49, or a salt thereof, wherein
the base structure B is represented by formula IIc: ##STR85##
wherein R and G are as defined in claim 49.
56. A compound according to claim 49, or a salt thereof, wherein
the base structure B is represented by the formula IId: ##STR86##
wherein R and G are as defined in claim 49.
57. A compound according to claim 49, or a salt thereof, wherein
the base structure B is represented by the formula IIe: ##STR87##
wherein R and G are as defined in claim 49.
58. A compound according to claim 49, or a salt thereof, wherein
R.sup.1, R.sup.2, R.sup.2', R.sup.3, R.sup.3', and R.sup.4 are
independently selected from the group consisting of H, halogen, OH,
N.sub.3, CN, optionally substituted alkyl, optionally substituted
alkenyl, optionally substituted alkynyl, optionally substituted
aryl, optionally substituted acyl, optionally substituted
arylalkyl, optionally substituted heterocyclyl, optionally
substituted heteroaryl, optionally substituted alkyloxy, optionally
substituted acyloxy, optionally substituted oxyacyl, optionally
substituted amino, optionally substituted aminoacyl, optionally
substituted aminoacyloxy, optionally substituted acylamino,
optionally substituted thio, or R.sup.2 and R.sup.2' together or
R.sup.3 and R.sup.3' together represents .dbd.O, .dbd.S, or =L-Y'
where L is N, CH, CF, CCl or CBr and Y' is H, halogen, N.sub.3,
methyl, ethyl or CN.
59. A compound according to claim 49, or a salt thereof, wherein
R.sup.1, R.sup.2, R.sup.2', R.sup.3, R.sup.3', and R.sup.4 are
independently selected from the group consisting of H, F, Cl, Br,
I, OH, SH, NH.sub.2, NHOH, NHNH.sub.2, N.sub.3, COOH, CN,
CONH.sub.2, C(S)NH.sub.2, COOR, R, OR, SR, SSR, NHR, and NR.sub.2
wherein at least one of R.sup.2 or R.sup.2' is a substituent other
than H, and wherein R is selected from the group consisting of
optionally substituted alkyl, optionally substituted alkenyl,
optionally substituted alkynyl, optionally substituted aryl,
optionally substituted acyl, and optionally substituted
arylalkyl.
60. A compound according to claim 49, or a salt thereof, wherein at
least one of R.sup.2 and R.sup.2' is methyl, hydroxyl or F.
61. A compound according to claim 49, or a salt thereof, wherein L'
is --CH.sub.2--.
62. A compound according to claim 49, or salt thereof, wherein
R.sup.4' is selected from --CH.sub.2--OH, --CF.sub.2OH,
--CCl.sub.2--OH, --C(CH.sub.3)(CH).sub.3OH, --CH(CH.sub.3)OH,
--CH.sub.2--CH.sub.2--P(O)(OH).sub.2,
--CH.sub.2--CH.sub.2--P(O)(OH).sub.2, --CH.sub.2SP(O)(OH).sub.2,
--CH.sub.2SH, --CF.sub.2SH, and
--CH.sub.2--O--P(O)(OPh)(NHCH(CH.sub.3)(CO.sub.2Me)).
63. A compound according to claim 62 or a salt thereof wherein
R.sup.4' is --CH.sub.2--OH.
64. A compound according to claim 49, or salt thereof, wherein the
sugar moiety is selected from the following formulae: ##STR88## or
C-5 monophosphate, diphosphate and triphosphate derivatives
thereof, or C-5 mono, di or triphosphate mimics, wherein R is as
defined in claim 49.
65. A compound according to claim 49, or salt thereof, wherein the
sugar moiety is selected from the following formulae: ##STR89## or
C-5 monophosphate, diphosphate and triphosphate derivatives
thereof, or C-5 mono, di or triphosphate mimics, wherein R is as
defined in claim 49.
66. A compound of formula (I) selected from the following formulae,
or a salt thereof: ##STR90## ##STR91## ##STR92## wherein: each R on
the sugar moiety is independently selected from H, optionally
substituted alkyl, optionally substituted alkenyl, optionally
substituted alkynyl, optionally substituted aryl, optionally
substituted acyl and optionally substituted arylalkyl; each R on
the base moiety is independently selected from H, optionally
substituted alkyl, optionally substituted aryl, optionally
substituted heteroaryl, optionally substituted heterocyclyl, and
optionally substituted arylalkyl; each G is independently selected
from H, halogen, CF.sub.3, optionally substituted alkyl, optionally
substituted aryl, optionally substituted heteroaryl, optionally
substituted heterocyclyl, and optionally substituted arylalkyl; and
C-5 monophosphate, diphosphate and triphosphate derivatives
thereof, or C-5 mono, di or triphosphate mimics.
67. A compound according to claim 49, or a salt thereof, wherein
the sugar moiety is represented by the formula: ##STR93## or C-5
monophosphate, diphosphate and triphosphate derivatives thereof, or
C-5 mono, di or triphosphate mimics, wherein R is as defined in
claim 49.
68. A compound according to claim 49, or a salt thereof, wherein
the sugar moiety is represented by the formula: ##STR94## or C-5
monophosphate, diphosphate and triphosphate derivatives thereof, or
C-5 mono, di or triphosphate mimics.
69. A compound according to claim 49, or a salt thereof, which is a
B anomer.
70. A compound according to claim 49, or a salt thereof, selected
from ##STR95## ##STR96## ##STR97##
71. A pharmaceutical composition comprising a compound according to
claim 49 or a salt thereof, and at least one pharmaceutically
acceptable carrier or diluent.
72. A pharmaceutical composition according to claim 71 for use in
the treatment of a viral infection.
73. A pharmaceutical composition according to claim 72, wherein the
virus is the hepatitis C virus.
74. A pharmaceutical composition according to claim 71 further
comprising one or more antiviral or antibacterial agents.
75. A pharmaceutical composition according to claim 74, wherein the
antiviral agents are selected from the group selected from
interferon and interferon derivatives, IMPDH inhibitors, antiviral
nucleosides, polymerase inhibitors and protease inhibitors.
76. A pharmaceutical composition according to claim 75, wherein the
composition comprises interferon or ribavirin.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to the use of bicyclic
nucleosides and nucleotides for the treatment of infectious
diseases, including viral infections, novel bicyclic nucleosides
and nucleotides and methods for their manufacture.
BACKGROUND OF THE INVENTION
[0002] Viral infections are a major threat to human health and
account for many serious infectious diseases. The most notable
viruses are the blood-borne viruses (BBV), which include hepatitis
C virus (HCV), hepatitis B virus (HBV) and human immunodeficiency
virus (HIV) which are all linked by their mode of transmission,
i.e., through blood or bodily fluids.
[0003] The Flaviviridae is a group of positive single-stranded RNA
viruses with a genome size from 9-15 kb. The Flaviviridae consists
of various genera including:
[0004] 1. Flaviviruses: This genus includes the Dengue virus,
Japanese Tick-Borne and the Yellow Fever virus. Apart from these
major groups, there are some additional Flaviviruses that are
unclassified.
[0005] 2. Hepaciviruses: This genus contains only one species, the
Hepatitis C virus (HCV), which is composed of many genotypes and
subtypes.
[0006] HCV is a major cause of viral hepatitis and has infected
more than 200 million people worldwide. Current treatment for HCV
infection is restricted to immunotherapy with interferon-.alpha.
alone or in combination with ribavirin, a nucleoside analog. This
treatment is effective in only about half the patient population.
Therefore, there is an urgent need for new HCV drugs. Hepatitis C
virus comprises a positive-strand RNA genome enclosed in a
nucleocapsid and lipid envelope and consists of approximately 9600
ribonucleotides, which encodes a polyprotein of about 3000 amino
acids (Dymock et al. Antiviral Chemistry & Chemotherapy 2000,
11, 79). A HCV protein, NS5B, released from the polyprotein,
possesses polymerase activity and is involved in the synthesis of
double-stranded RNA from the single-stranded viral RNA genome that
serves as the template. The reproduction of HCV virus may be
prevented through the manipulation of NS5B's polymerase activity.
The inhibition of NS5B protein would suppress or prevent the
formation of the double-stranded HCV RNA. Alternatively, a
nucleoside analog also may be incorporated into the extending RNA
strand and act as a chain-terminator. Furthermore, a deteriorating
nucleoside analog also may be incorporated into the extending RNA,
which may cause mutagenic damage to the viral genome. Recently,
several PCT patent applications (WO 99/43691, WO 01/32153, WO
01/60315, WO 01/79246, WO 01/90121, WO 01/92282, WO 02/18404, WO
02/057287, WO 02/057425) have described nucleoside analogs as
anti-HCV agents in in vitro assays.
[0007] HBV has acutely infected almost a third of the world's human
population, and about 5% of the infected are chronic carriers of
the virus (Delaney I V et al., Antiviral Chemistry &
Chemotherapy 2001, 12, 1-35). Chronic HBV infection causes liver
damage that frequently progresses to cirrhosis and/or liver cancer
later in the life. Despite the availability and widespread use of
effective vaccines and chemotherapy, the number of chronic carriers
approaches 400 million worldwide. Therefore, more effective
anti-HBV drugs need to be developed.
[0008] HIV causes progressive degeneration of the immune system,
leading to the development of AIDS. A number of drugs have been
used clinically, including reverse transcriptase inhibitors and
protease inhibitors. Currently, combination therapies are used
widely for the treatment of AIDS in order to reduce the drug
resistance. Despite the progress in the development of anti-HIV
drugs, AIDS is still one of the leading epidemic diseases.
[0009] Apart from the BBV's discussed above certain other acute
viral infections also impose a great threat to human life,
including the HSV, CMV, influenza viruses, West Nile virus, SARS
virus, small pox, EBV, VZV and RSV. Accordingly, this highlights
the continued need for the development of different antiviral
drugs.
[0010] Bacterial infections have long been the sources of many
infectious diseases. The widespread use of antibiotics has produced
many new strains of life-threatening antibiotic resistant bacteria.
Fungal infections are another type of infectious diseases, some of
which also can be life-threatening. There is an ever increasing
demand for the treatment of bacterial and fungal infections. As
such, antimicrobial drugs based on new mechanisms of action are
especially important.
[0011] Nucleoside drugs have been used clinically for decades for
the treatment of viral infections and proliferative disorders such
as cancer. Most of the nucleoside drugs are classified as
antimetabolites. After they enter cells, nucleoside analogs are
phosphorylated successively to nucleoside 5'-monophosphates,
5'-diphosphates, and 5'-triphosphates. In most cases, nucleoside
triphosphates, e.g., 3'-azido-3'-deoxythymidine triphosphate (AZT,
an anti-HIV drug) and arabinosylcytosine triphosphate (cytarabine,
an anticancer drug), are the active chemical entities that inhibit
DNA or RNA synthesis, through a competitive inhibition of
polymerases and subsequent incorporation of modified nucleotides
into DNA or RNA sequences. In a few cases, nucleoside analogs exert
effects at lower phosphate levels. For instance,
5-fluoro-2'-deoxyuridine 5'-monophosphate (an anticancer drug) and
2',2'-difluoro-2'-deoxycytidine 5'-diphosphate (an anticancer drug)
have been shown to inhibit thymidylate synthase and ribonucleotide
reductase, respectively. Although nucleoside analogs themselves may
act at the nonphosphate level such as the inhibitors of adenosine
kinases and the ligands of adenosine receptors, currently,
clinically-useful nucleoside drugs primarily depend on cellular
activation by nucleoside kinases and nucleotide kinases.
[0012] At least, two criteria are pertinent for nucleoside
antiviral drugs: 1) nucleoside analogs should anabolise to
nucleotides in cells; and 2) the anabolised nucleotides should
target selectively viral enzymes. In order to be phosphorylated in
cells and selectively target preferred enzymes, nucleoside analogs
should have favourable modifications on their sugar and base
moieties. To obtain such favourable nucleoside analogs, a general
approach is to generate diverse nucleoside analogs by modifying the
base or the sugar, or by modifying both base and sugar moieties.
Numerous examples exist in the literature for the synthesis of a
variety of modified nucleosides (Chemistry of Nucleosides and
Nucleotides Vol. 1 (1988), Vol. 2 (1991), Vol. 3 (1994), edited by
L. B. Townsend, Plenum Press; Handbook of Nucleoside Synthesis by
H. Vorbruggen and C. Ruh-Pohlenz, John Wiley & Sons, Inc.,
2001; The Organic Chemistry of Nucleic Acids by Y. Mizuno,
Elsevier, 1986).
[0013] However, there are certain classes of nucleoside compounds
that were not explored intensively for their antiviral activities
before the present invention. A class of such compounds is bicyclic
nucleosides which are not derived from purine bases. Disclosures of
bicyclic nucleosides are very limited considering that natural
adenine and guanine (purines) based ribonucleotides and deoxy
derivatives thereof, have bicyclic base moieties. WO 01/92282 A2,
WO 01/90121 A2 and WO 04/058792 disclose derivatives of purine
nucleosides. In contrast to these publications, the present
invention discloses that a certain new class of bicyclic
nucleosides and nucleotides display biological activity which may
be particularly useful for the treatment of infectious diseases,
including viral infections.
SUMMARY OF THE INVENTION
[0014] The present invention relates to a particular class of
bicyclic nucleosides, nucleotides, and derivatives thereof and
their use in the treatment of microbial infections, and
specifically viral infections.
[0015] In particular, the present invention provides a method for
the treatment of a microbial infection, comprising administering an
effective amount of a compound of the formula (I) or a
pharmaceutically acceptable salt thereof; ##STR1## wherein:
[0016] A is O, S, CH.sub.2, CHF, CF.sub.2 or NR;
[0017] R.sup.1, R.sup.2, R.sup.2', R.sup.3, R.sup.3', and R.sup.4
are independently selected from the group consisting of H, halogen,
OH, N.sub.3, CN, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted aryl, optionally substituted acyl, optionally
substituted arylalkyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, optionally substituted
cycloalkyl, optionally substituted cycloalkenyl, optionally
substituted alkyloxy, optionally substituted alkenyloxy, optionally
substituted alkynoxy, optionally substituted aryloxy, optionally
substituted acyloxy, optionally substituted oxyacyl, optionally
substituted arylalkoxy, optionally substituted heterocycloxy,
optionally substituted heteroaryloxy, optionally substituted
cycloalkoxy, optionally substituted cycloalkenoxy, optionally
substituted amino, optionally substituted aminoacyl, optionally
substituted aminoacyloxy, optionally substituted acylamino,
optionally substituted oxyacylamino, optionally substituted
oxyacyloxy, optionally substituted acylimino, optionally
substituted acyliminoxy, optionally substituted oxyacylimino,
optionally substituted aminothioacyl, optionally substituted
thioacylamino, optionally substituted aminosulfinyl, optionally
substituted aminosulfonyl, optionally substituted thio, optionally
substituted thioacyl, optionally substituted thioacyloxy,
optionally substituted oxythioacyl, optionally substituted
oxythioacyloxy, optionally substituted phosphorylamino, optionally
substituted sulfinyl, optionally substituted sulfonyl, optionally
substituted sulfinylamino, optionally substituted sulfonylamino,
optionally substituted oxysulfinylamino, and optionally substituted
oxysulfonylamino, or R.sup.2 and R.sup.2' together or R.sup.3 and
R.sup.3' together represents .dbd.O, .dbd.S, or =L-Y' where L is N,
CH, CF, CCl or CBr and Y' is H, halogen, N.sub.3, methyl, ethyl or
CN;
[0018] R.sup.4' is --CY.sub.2SH, --CY.sub.2OH, --CY.sub.2NH.sub.2,
or -L'-R.sup.5;
[0019] L' is selected from the group consisting of --CY.sub.2--,
--CY.sub.2CY.sub.2--, --CY.sub.2OCY.sub.2--, --CY.sub.2SCY.sub.2--
and --CY.sub.2NHCY.sub.2--;
[0020] each Y is independently selected from the group consisting
of H, F, Cl, Br, OR, C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl
and C.sub.2-C.sub.6alkynyl, wherein C.sub.1-C.sub.6alkyl,
C.sub.2-C.sub.6alkenyl, and C.sub.2-C.sub.6alkynyl may be
optionally substituted with one or more groups selected from F, Cl,
Br, OH, COOH, COOCH.sub.3, SH, SCH.sub.3, NH.sub.2, NHCH.sub.3,
N(CH.sub.3).sub.2, CN, NO.sub.2, C(O)NH.sub.2, C(O)NHCH.sub.3,
N.sub.3, C(S)NH.sub.2, OCH.sub.3, and OCH.sub.2CH.sub.3;
[0021] R.sup.5 is selected from the group consisting of OR,
NR.sub.2, monophosphate, diphosphate, and triphosphate, or a mono,
di or triphosphate mimic;
[0022] each R is independently selected from the group consisting
of H, CF.sub.3, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted aryl, optionally substituted acyl, optionally
substituted cycloalkyl, optionally substituted cycloalkenyl,
optionally substituted heteroaryl, optionally substituted
heterocyclyl, and optionally substituted arylalkyl;
[0023] B is a group of formula (II) ##STR2##
[0024] wherein, if Z is a participant in a .pi. bond (double bond),
Z is independently selected from N or C-G; or, if Z is not a
participant in a .pi. bond (double bond), Z is independently
selected from O, S, Se, NR, NOR, NNR.sub.2, CO, CS, CNR, SO,
S(O).sub.2, SeO, Se(O).sub.2 or C(G).sub.2, wherein each G is
independently selected from the group consisting of H, halogen, OR,
SR, NR.sub.2, NROR, N.sub.3, COOR, CN, CONR.sub.2, C(S)NR.sub.2,
C(.dbd.NR)NR.sub.2, and R; and
[0025] where any two adjacent Z are not both selected from O, S,
and Se, or not both selected from CO, CS, CNR, SO, S(O).sub.2, SeO,
and Se(O).sub.2;
[0026] wherein, if X is a participant in a .pi. bond (double bond),
X is C; or if X is not a participant in a .pi. bond (double bond),
X is CR or N;
[0027] wherein, if R'' is a participant in a .pi. bond (double
bond), R'' is O, S, Se, NR, NOR or NNR.sub.2; or if R'' is not a
participant in a .pi. bond (double bond), R'' is OR, SR, F, Cl, R,
or SeR; and
[0028] dashed lines (---) indicate a possible .pi. or double
bond;
[0029] optionally in combination with one or more antiviral or
antibacterial agents.
[0030] The present invention also further provides the use of a
compound of formula (I) or a salt thereof, ##STR3## wherein:
[0031] A is O, S, CH.sub.2, CHF, CF.sub.2 or NR;
[0032] R.sup.1, R.sup.2, R.sup.2', R.sup.3, R.sup.3', and R.sup.4
are independently selected from the group consisting of H, halogen,
OH, N.sub.3, CN, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted aryl, optionally substituted acyl, optionally
substituted arylalkyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, optionally substituted
cycloalkyl, optionally substituted cycloalkenyl, optionally
substituted alkyloxy, optionally substituted alkenyloxy, optionally
substituted alkynoxy, optionally substituted aryloxy, optionally
substituted acyloxy, optionally substituted oxyacyl, optionally
substituted arylalkoxy, optionally substituted heterocycloxy,
optionally substituted heteroaryloxy, optionally substituted
cycloalkoxy, optionally substituted cycloalkenoxy, optionally
substituted amino, optionally substituted aminoacyl, optionally
substituted aminoacyloxy, optionally substituted acylamino,
optionally substituted oxyacylamino, optionally substituted
oxyacyloxy, optionally substituted acylimino, optionally
substituted acyliminoxy, optionally substituted oxyacylimino,
optionally substituted aminothioacyl, optionally substituted
thioacylamino, optionally substituted aminosulfinyl, optionally
substituted aminosulfonyl, optionally substituted thio, optionally
substituted thioacyl, optionally substituted thioacyloxy,
optionally substituted oxythioacyl, optionally substituted
oxythioacyloxy, optionally substituted phosphorylamino, optionally
substituted sulfinyl, optionally substituted sulfonyl, optionally
substituted sulfinylamino, optionally substituted sulfonylamino,
optionally substituted oxysulfinylamino, and optionally substituted
oxysulfonylamino, or R.sup.2 and R.sup.2' together or R.sup.3 and
R.sup.3' together represents .dbd.O, .dbd.S, or =L-Y' where L is N,
CH, CF, CCl or CBr and Y' is H, halogen, N.sub.3, methyl, ethyl or
CN;
[0033] R.sup.4' is --CY.sub.2SH, --CY.sub.2OH, --CY.sub.2NH.sub.2,
or -L-R.sup.5;
[0034] L' is selected from the group consisting of --CY.sub.2--,
--CY.sub.2CY.sub.2--, --CY.sub.2OCY.sub.2--, --CY.sub.2SCY.sub.2--
and --CY.sub.2NHCY.sub.2--;
[0035] each Y is independently selected from the group consisting
of H, F, Cl, Br, OR, C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl
and C.sub.2-C.sub.6alkynyl, wherein C.sub.1-C.sub.6alkyl,
C.sub.2-C.sub.6alkenyl, and C.sub.2-C.sub.6alkynyl may be
optionally substituted with one or more groups selected from F, Cl,
Br, OH, COOH, COOCH.sub.3, SH, SCH.sub.3, NH.sub.2, NHCH.sub.3,
N(CH.sub.3).sub.2, CN, NO.sub.2, C(O)NH.sub.2, C(O)NHCH.sub.3,
N.sub.3, C(S)NH.sub.2, OCH.sub.3, and OCH.sub.2CH.sub.3;
[0036] R.sup.5 is selected from the group consisting of OR,
NR.sub.2, monophosphate, diphosphate, and triphosphate, or a mono,
di or triphosphate mimic;
[0037] each R is independently selected from the group consisting
of H, CF.sub.3, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted aryl, optionally substituted acyl, optionally
substituted cycloalkyl, optionally substituted cycloalkenyl,
optionally substituted heteroaryl, optionally substituted
heterocyclyl, and optionally substituted arylalkyl;
[0038] B is a group of formula (II) ##STR4##
[0039] wherein, if Z is a participant in a .pi. bond (double bond),
Z is independently selected from N or C-G; or, if Z is not a
participant in a .pi. bond (double bond), Z is independently
selected from O, S, Se, NR, NOR, NNR.sub.2, CO, CS, CNR, SO,
S(O).sub.2, SeO, Se(O).sub.2 or C(G).sub.2, wherein each G is
independently selected from the group consisting of H, halogen, OR,
SR, NR.sub.2, NROR, N.sub.3, COOR, CN, CONR.sub.2, C(S)NR.sub.2,
C(.dbd.NR)NR.sub.2, and R; and
[0040] where any two adjacent Z are not both selected from O, S,
and Se, or not both selected from CO, CS, CNR, SO, S(O).sub.2, SeO,
and Se(O).sub.2;
[0041] wherein, if X is a participant in a 7 bond (double bond), X
is C; or if X is not a participant in a .pi. bond (double bond), X
is CR or N;
[0042] wherein, if R'' is a participant in a .pi. bond (double
bond), R'' is O, S, Se, NR, NOR or NNR.sub.2; or if R'' is not a
participant in a 71 bond (double bond), R'' is OR, SR, F, Cl, R, or
SeR; and
[0043] dashed lines (---) indicate a possible .pi. or double
bond;
[0044] in the manufacture of a medicament for the treatment of a
microbial infection.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0045] "Alkyl" refers to monovalent alkyl groups which may be
straight chained or branched and preferably have from 1 to 10
carbon atoms or more preferably 1 to 6 carbon atoms. Examples of
such alkyl groups include methyl, ethyl, n-propyl, iso-propyl,
n-butyl, iso-butyl, n-hexyl, and the like.
[0046] "Aryl" refers to an unsaturated aromatic carbocyclic group
having a single ring (e.g., phenyl) or multiple condensed rings
(e.g., naphthyl or anthryl), preferably having from 6 to 14 carbon
atoms. Examples of aryl groups include phenyl, naphthyl and the
like.
[0047] "Arylene" refers to a divalent aryl group wherein the aryl
group is as described above.
[0048] "Aryloxy" refers to the group aryl-O-- wherein the aryl
group is as described above.
[0049] "Arylalkyl" refers to -alkylene-aryl groups preferably
having from 1 to 10 carbon atoms in the alkylene moiety and from 6
to 10 carbon atoms in the aryl moiety. Such arylalkyl groups are
exemplified by benzyl, phenethyl and the like.
[0050] "Arylalkoxy" refers to the group arylalkyl-O-- wherein the
arylalkyl group are as described above. Such arylalkoxy groups are
exemplified by benzyloxy and the like.
[0051] "Alkoxy" refers to the group alkyl-O-- where the alkyl group
is as described above. Examples include, methoxy, ethoxy,
n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy,
n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.
[0052] "Alkenyl" refers to a monovalent alkenyl group which may be
straight chained or branched and preferably have from 2 to 10
carbon atoms and more preferably 2 to 6 carbon atoms and have at
least 1 and preferably from 1-2, carbon to carbon, double bonds.
Examples include ethenyl (--CH.dbd.CH.sub.2), n-propenyl
(--CH.sub.2CH.dbd.CH.sub.2), iso-propenyl
(--C(CH.sub.3).dbd.CH.sub.2), but-2-enyl
(--CH.sub.2CH.dbd.CHCH.sub.3), and the like.
[0053] "Alkenyloxy" refers to the group alkenyl-O-- wherein the
alkenyl group is as described above.
[0054] "Alkynyl" refers to alkynyl groups preferably having from 2
to 10 carbon atoms and more preferably 2 to 6 carbon atoms and
having at least 1, and preferably from 1-2, carbon to carbon,
triple bonds. Examples of alkynyl groups include ethynyl
(--C.ident.CH), propargyl (--CH.sub.2C.ident.CH), pent-2-ynyl
(--CH.sub.2C.ident.CCH.sub.2--CH.sub.3), and the like.
[0055] "Alkynyloxy" refers to the group alkynyl-O-- wherein the
alkynyl groups are described above.
[0056] "Acyl" refers to groups H--C(O)--, alkyl-C(O)--,
cycloalkyl-C(O)--, aryl-C(O)--, heteroaryl-C(O)-- and
heterocyclyl-C(O)--, where alkyl, cycloalkyl, aryl, heteroaryl and
heterocyclyl are as described herein.
[0057] "Oxyacyl" refers to groups H--OC(O)--, alkyl-OC(O)--,
cycloalkyl-OC(O)--, aryl-OC(O)--, heteroaryl-OC(O)--, and
heterocyclyl-OC(O)--, where alkyl, cycloalkyl, aryl, heteroaryl and
heterocyclyl are as described herein.
[0058] "Amino" refers to the group --NR'''R''' where each R''' is
independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, and
heterocyclyl and where alkyl, cycloalkyl, aryl, heteroaryl and
heterocyclyl is as described herein.
[0059] "Aminoacyl" refers to the group --C(O)NR'''R''' where each
R''' is independently hydrogen, alkyl, cycloalkyl, aryl,
heteroaryl, and heterocyclyl and where alkyl, cycloalkyl, aryl,
heteroaryl and heterocyclyl are as described herein.
[0060] "Acylamino" refers to the group --NR'''C(O)R''' where each
R''' is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl
and heterocyclyl and where alkyl, cycloalkyl, aryl, heteroaryl, and
heterocyclyl are as described herein.
[0061] "Acyloxy" refers to the groups --OC(O)--H, --OC(O)-alkyl,
--OC(O)-aryl, --C(O)O-heteroaryl, and --C(O)O-heterocyclyl where
alkyl, aryl, heteroaryl and heterocyclyl are as described
herein.
[0062] "Aminoacyloxy" refers to the groups --OC(O)NR'''--H,
--OC(O)NR'''-alkyl, --OC(O)NR'''-aryl, --OC(O)NR'''-heteroaryl, and
--OC(O)NR'''-heterocyclyl where R''' is independently hydrogen,
alkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl and where
alkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl are as
described herein.
[0063] "Oxyacylamino" refers to the groups --NR'''OC(O)OH,
--NR'''OC(O)O-alkyl, --NR'''OC(O)O-aryl, --NR'''C(O)O-heteroaryl,
and NR'''OC(O)O-heterocyclyl where R''' is independently hydrogen,
alkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl and where
alkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl are as
described herein.
[0064] "Oxyacyloxy" refers to the groups --OC(O)--OH,
--OC(O)O-alkyl, --O--C(O)O-aryl, --OC(O)O-heteroaryl, and
--OC(O)O-heterocyclyl where alkyl, cycloalkyl, aryl, heteroaryl,
and heterocyclyl are as described herein.
[0065] "Acylimino" refers to the groups --C(NR''')--R''' where each
R''' is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl
and heterocyclyl and where alkyl, cycloalkyl, aryl, heteroaryl, and
heterocyclyl are as described herein.
[0066] "Acyliminoxy" refers to the groups --O--C(NR''')--R''' where
each R''' is independently hydrogen, alkyl, cycloalkyl, aryl,
heteroaryl and heterocyclyl and where alkyl, cycloalkyl, aryl,
heteroaryl, and heterocyclyl are as described herein.
[0067] "Oxyacylimino" refers to the groups --C(NR''')--OR''' where
each R''' is independently hydrogen, alkyl, cycloalkyl, aryl,
heteroaryl and heterocyclyl and where alkyl, cycloalkyl, aryl,
heteroaryl, and heterocyclyl are as described herein.
[0068] "Cycloalkyl" refers to cyclic alkyl groups having a single
cyclic ring or multiple condensed rings, preferably incorporating 3
to 8 carbon atoms. Such cycloalkyl groups include, by way of
example, single ring structures such as cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cyclooctyl, and the like, or multiple ring
structures such as adamantanyl, and the like.
[0069] "Cycloalkenyl" refers to cyclic alkenyl groups having a
single cyclic ring and at least one point of internal unsaturation,
preferably incorporating 4 to 8 carbon atoms. Examples of suitable
cycloalkenyl groups include, for instance, cyclobut-2-enyl,
cyclopent-3-enyl, cyclohex-4-enyl, cyclooct-3-enyl and the
like.
[0070] "Halo" or "halogen" refers to fluoro, chloro, bromo and
iodo.
[0071] "Heteroaryl" refers to a monovalent aromatic heterocyclic
group which fulfils the Huckel criteria for aromaticity (i.e.,
contains 4n+2.pi. electrons, is planar and conjugated) and
preferably has from 2 to 10 carbon atoms and 1 to 4 heteroatoms
selected from oxygen, nitrogen, selenium, and sulfur within the
ring (and includes oxides of sulfur, selenium and nitrogen). Such
heteroaryl groups can have a single ring (e.g., pyridyl, pyrrolyl
or N-oxides thereof or furyl) or multiple condensed rings (e.g.,
indolizinyl, benzoimidazolyl, coumarinyl, quinolinyl, isoquinolinyl
or benzothienyl).
[0072] "Heterocyclyl" refers to a monovalent saturated or
unsaturated group having a single ring or multiple condensed rings,
preferably from 1 to 8 carbon atoms and from 1 to 4 hetero atoms
selected from nitrogen, sulfur, oxygen, selenium or phosphorous
within the ring. The most preferred heteroatom is nitrogen.
[0073] Examples of heterocyclyl and heteroaryl groups include, but
are not limited to, oxazole, pyrrole, imidazole, pyrazole,
pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole,
indole, indazole, purine, quinolizine, isoquinoline, quinoline,
phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline,
pteridine, carbazole, carboline, phenanthridine, acridine,
phenanthroline, isothiazole, phenazine, isoxazole, isothiazole,
phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine,
piperazine, indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline,
4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiadiazoles,
oxadiazole, oxatriazole, tetrazole, thiazolidine, thiophene,
benzo[b]thiophene, morpholino, piperidinyl, pyrrolidine,
tetrahydrofuranyl, triazole, and the like.
[0074] "Thio" refers to groups H--S--, alkyl-S--, cycloalkyl-S--,
aryl-S--, heteroaryl-S--, and heterocyclyl-S--, where alkyl,
cycloalkyl, aryl, heteroaryl and heterocyclyl are as described
herein.
[0075] "Thioacyl" refers to groups H--C(S)--, alkyl-C(S)--,
cycloalkyl-C(S)--, aryl-C(S)--, heteroaryl-C(S)--, and
heterocyclyl-C(S)--, where alkyl, cycloalkyl, aryl, heteroaryl and
heterocyclyl are as described herein.
[0076] "Oxythioacyl" refers to groups HO--C(S)--, alkylO--C(S)--,
cycloalkylO--C(S)--, arylO--C(S)--, heteroarylO--C(S)--, and
heterocyclylO--C(S)--, where alkyl, cycloalkyl, aryl, heteroaryl
and heterocyclyl are as described herein.
[0077] "Oxythioacyloxy" refers to groups HO--C(S)--O--,
alkylO--C(S)--O--, cycloalkylO--C(S)--O--, arylO--C(S)--O--,
heteroarylO--C(S)--O--, and heterocyclylO--C(S)--O--, where alkyl,
cycloalkyl, aryl, heteroaryl and heterocyclyl are as described
herein.
[0078] "Phosphorylamino" refers to the groups
--NR'''--P(O)(R'''')(OR''''') where R''' represents H, alkyl,
cycloalkyl, alkenyl, or aryl, R'''' represents OR''''' or is
hydroxy or amino and R''''' is alkyl, cycloalkyl, aryl or
arylalkyl, where alkyl, amino, alkenyl, aryl, cycloalkyl, and
arylalkyl are as described herein.
[0079] "Thioacyloxy" refers to groups H--C(S)--O--,
alkyl-C(S)--O--, cycloalkyl-C(S)--O--, aryl-C(S)--O--,
heteroaryl-C(S)--O--, and heterocyclyl-C(S)--O--, where alkyl,
cycloalkyl, aryl, heteroaryl, and heterocyclyl are as described
herein.
[0080] "Sulfinyl" refers to groups H--S(O)--, alkyl-S(O)--,
cycloalkyl-S(O)--, aryl-S(O)--, heteroaryl-S(O)--, and
heterocyclyl-S(O)--, where alkyl, cycloalkyl, aryl, heteroaryl and
heterocyclyl are as described herein.
[0081] "Sulfonyl" refers to groups H--S(O).sub.2--,
alkyl-S(O).sub.2--, cycloalkyl-S(O).sub.2--, aryl-S(O).sub.2--,
heteroaryl-S(O).sub.2--, and heterocyclyl-S(O).sub.2--, where
alkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl are as
described herein.
[0082] "Sulfinylamino" refers to groups H--S(O)--NR'''--,
alkyl-S(O)--NR'''--, cycloalkyl-S(O)--NR'''--, aryl-S(O)--NR'''--,
heteroaryl-S(O)--NR'''--, and heterocyclyl-S(O)--NR'''--, where
R''' is independently hydrogen, alkyl, cycloalkyl, aryl,
heteroaryl, and heterocyclyl and where alkyl, cycloalkyl, aryl,
heteroaryl and heterocyclyl are as described herein.
[0083] "Sulfonylamino" refers to groups H--S(O).sub.2--NR'''--,
alkyl-S(O).sub.2--NR'''--, cycloalkyl-S(O).sub.2--NR'''--,
aryl-S(O).sub.2--NR'''--, heteroaryl-S(O).sub.2--NR'''--, and
heterocyclyl-S(O).sub.2--NR'''--, where R''' is independently
hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl and
alkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl are as
described herein.
[0084] "Oxysulfinylamino" refers to groups HO--S(O)--NR'''--,
alkylO--S(O)--NR'''--, cycloalkylO--S(O)--NR'''--,
arylO--S(O)--NR'''--, heteroarylO--S(O)--NR'''--, and
heterocyclylO--S(O)--NR'''--, where R''' is independently hydrogen,
alkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl and where
alkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl are as
described herein.
[0085] "Oxysulfonylamino" refers to groups HO--S(O).sub.2--NR'''--,
alkylO--S(O).sub.2--NR'''--, cycloalkylO--S(O).sub.2--NR'''--,
arylO--S(O).sub.2--NR'''--, heteroarylO--S(O).sub.2--NR'''--, and
heterocyclylO--S(O).sub.2--NR'''--, where R''' is independently
hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl and
where alkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl are as
described herein.
[0086] "Aminothioacyl" refers to groups R'''R'''N--C(S)--, where
each R''' is independently hydrogen, alkyl, cycloalkyl, aryl,
heteroaryl, where heterocyclic and alkyl, cycloalkyl, aryl,
heteroaryl and heterocyclyl are as described herein.
[0087] "Thioacylamino" refers to groups H--C(S)--NR'''--,
alkyl-C(S)--NR'''--, cycloalkyl-C(S)--NR'''--, aryl-C(S)--NR'''--,
heteroaryl-C(S)--NR'''--, and heterocyclyl-C(S)--NR'''--, where
R''' is independently hydrogen, alkyl, cycloalkyl, aryl,
heteroaryl, and where heterocyclyl and alkyl, cycloalkyl, aryl,
heteroaryl and heterocyclyl are as described herein.
[0088] "Aminosulfinyl" refers to groups R'''R'''N--S(O)--, where
each R''' is independently hydrogen, alkyl, cycloalkyl, aryl,
heteroaryl, and heterocyclic and where alkyl, cycloalkyl, aryl,
heteroaryl and heterocyclyl are as described herein.
[0089] "Aminosulfonyl" refers to groups R'''R'''N--S(O).sub.2--,
where each R''' is independently hydrogen, alkyl, cycloalkyl, aryl,
heteroaryl, and heterocyclic and where alkyl, cycloalkyl, aryl,
heteroaryl and heterocyclyl are as described herein.
[0090] In this specification "optionally substituted" is taken to
mean that a group may or may not be further substituted or fused
(so as to form a condensed polycyclic group) with one or more
groups selected from hydroxyl, acyl, alkyl, alkoxy, alkenyl,
alkenyloxy, alkynyl, alkynyloxy, amino, aminoacyl, thio, arylalkyl,
arylalkoxy, aryl, aryloxy, acylamino, cyano, halogen, nitro, sulfo,
phosphono, phosphorylamino, phosphinyl, heteroaryl, heteroaryloxy,
heterocyclyl, heterocycloxy, oxyacyl, acyloxy, oxime, oxime ether,
hydrazone, oxyacylamino, aminoacyloxy, trihalomethyl,
trialkylsilyl, pentafluoroethyl, trifluoromethoxy, difluoromethoxy,
trifluoromethanethio, trifluoroethenyl, mono- and di-alkylamino,
mono- and di-(substituted alkyl)amino, mono- and di-arylamino,
mono- and di-heteroarylamino, mono- and di-heterocyclyl amino, and
unsymmetric di-substituted amines having different substituents
selected from alkyl, aryl, heteroaryl and heterocyclyl, and the
like. For instance, an "optionally substituted amino" group may
include amino acid and peptide residues.
[0091] The term "base", unless otherwise specified, refers to the
base moiety of a nucleoside or nucleotide. The base moiety is the
nitrogen-heterocycle portion of a nucleoside or nucleotide. The
base moiety of a nucleotide of formula (I) is a bicyclic
heterocycle represented by formula (II) and designated "B". The
nucleoside base is attached to the sugar moiety of a nucleoside in
such ways that both .alpha. and .beta. anomers of D or L
nucleosides can be produced. This is denoted by use of the bond
which links the base to the sugar moiety.
[0092] The term "sugar" refers to the furanose portion of a
nucleoside. The sugar moiety of formula (I) nucleosides,
nucleotides and nucleotides mimics and/or prodrugs thereof may
contain one or more substituents at their C1-, C2-, C3- and
C4-position of the furanose. Substituents may be directed to either
the .alpha.- or .beta.-face of the furanose. The nucleoside base
can be considered as a substituent at the C-1 position of the
furanose and is preferably directed to the .beta.-face of the
sugar. The .beta.-face is the side of a furanose on which a purine
or pyrimidine base of natural .beta.-D-nucleosides is present. The
.alpha.-face is the side of the sugar opposite to the
.beta.-face.
[0093] Examples of a "protecting group" for O, S, or N moieties
such as hydroxy or NH.sub.2, includes acyl groups, silyl groups,
and the like. Suitable protecting groups for these and other
moieties are described by T. W., Greene and P. G. M. Wuts;
Protecting Groups in Organic Synthesis, 3.sup.rd Ed, John Wiley
& Sons, Inc. (1999), incorporated herein by reference.
[0094] The term "infection" or "microbial infection" refers to the
infection caused by an infectious agent or microbe, such as
bacteria, parasite (including protozoan), virus or fungus
(including unicellular and multicellular). Examples of microbes
that cause such infection include: Acanthamoeba, African Sleeping
Sickness (Trypanosomiasis), amebiasis, American Trypanosomiasis
(Chagas Disease), Bilharzia (Schistosomiasis), cryptosporidiosis
(diarrheal disease, Cryptosporidium Parvum), Giardiasis (diarrheal
disease, Giardia lamblia), hepatitis A, B, C, D, E, leishmaniasis
(skin sores and visceral), malaria (Plasmodium falciparum),
Salmonella enteritides infection (stomach cramps, diarrhea and
fever), tuberculosis (mycobacterium tuberculosis), varicella
(chicken pox), yellow fever, pneumonias, urinary tract infections
(Chlamydia and Mycoplasma), meningitis and meningococcal
septicemia, skin and soft tissue infections (Staphylococcus
aureus), lower respiratory tract infections (bacterial pathogens or
viral pathogens).
[0095] Common infections caused by microbes are further outlined in
the following chart: TABLE-US-00001 Infection Bacteria Fungus
Protozoa Virus AIDS X Athlete's Foot X Chicken Pox X Common Cold X
Diarrheal Disease X X X Dengue X Flu X Genital Herpes X Malaria X X
Meningitis X Pneumonia X X Sinusitis X X Skin Disease X X X X Strep
Throat X Tuberculosis X Urinary Tract Infections X Vaginal
Infections X X Viral Hepatitis X
[0096] In relation to the therapeutic methods of the present
invention the compounds of formula (I) may be particularly useful
for treating a microbial infection which is a viral infection
caused by an RNA virus, such as a virus belonging to group
Flaviviridae, for instance Flaviviruses or HCV, or a DNA or
retrovirus such as HBV or HIV. In a preferred embodiment the method
of the present invention treats a viral infection caused by an RNA
virus of the group Flaviviridae and in particular HCV.
[0097] The compounds of formula (I) are administered to the subject
in a therapeutic effective amount. As used herein, a therapeutic
effective amount is intended to include at least partially
attaining the desired effect, or delaying the onset of, or
inhibiting the progression of, or halting or reversing altogether
the onset or progression of the particular disease of condition
being treated.
[0098] As used herein, the term "effective amount" relates to an
amount of compound which, when administered according to a desired
dosing regimen, provides the desired therapeutic activity. Dosing
may occur at intervals of minutes, hours, days, weeks, months or
years or continuously over any one of these periods. Suitable
dosages lie within the range of about 0.1 ng per kg of body weight
to 10 g per kg of body weight per dosage. The dosage may be in the
range of 1 .mu.g to 10 g per kg of body weight per dosage, such as
is in the range of 1 mg to 10 g per kg of body weight per dosage.
In one embodiment, the dosage may be in the range of 1 mg to 500 mg
per kg of body weight per dosage. In another embodiment, the dosage
may be in the range of 1 mg to 250 mg per kg of body weight per
dosage. In yet another preferred embodiment, the dosage may be in
the range of 1 mg to 100 mg per kg of body weight per dosage, such
as up to 50 mg per body weight per dosage.
[0099] Suitable dosage amounts and dosing regimens can be
determined by the attending physician and may depend on the
particular condition being treated, the severity of the condition
as well as the general age, health and weight of the subject.
[0100] The active ingredient may be administered in a single dose
or a series of doses. While it is possible for the active
ingredient to be administered alone, it is preferable to present it
as a composition, preferably as a pharmaceutical composition. The
formulation of such compositions is well known to those skilled in
the art. The composition may contain any suitable carriers,
diluents or excipients. These include all conventional solvents,
dispersion media, fillers, solid carriers, coatings, further
antifungal and antibacterial agents, dermal penetration agents,
surfactants, isotonic and absorption agents and the like. It will
be understood that the compositions of the invention may also
include other supplementary physiologically active agents.
[0101] The carrier must be pharmaceutically "acceptable" in the
sense of being compatible with the other ingredients of the
composition and not injurious to the subject. Compositions include
those suitable for oral, rectal, nasal, topical (including buccal
and sublingual), vaginal or parental (including subcutaneous,
intramuscular, intravenous and intradermal) administration. The
compositions may conveniently be presented in unit dosage form and
may be prepared by any methods well known in the art of pharmacy.
Such methods include the step of bringing into association the
active ingredient with the carrier which constitutes one or more
accessory ingredients. In general, the compositions are prepared by
uniformly and intimately bringing into association the active
ingredient with liquid carriers or finely divided solid carriers or
both, and then if necessary shaping the product.
[0102] Compositions suitable for oral administration may be
presented as discrete units such as capsules, sachets 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
presented as a bolus, electuary or paste.
[0103] A tablet may be made by compression or moulding, 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., inert diluent, preservative disintegrant
(e.g., sodium starch glycolate, cross-linked polyvinyl pyrrolidone,
cross-linked sodium carboxymethyl cellulose) surface-active or
dispersing agent. Moulded tablets may be made by moulding 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,
hydroxypropylmethyl cellulose 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.
[0104] Compositions suitable for topical administration in the
mouth include lozenges comprising the active ingredient in a
flavoured base, usually sucrose and acacia or tragacanth gum;
pastilles comprising the active ingredient in an inert basis such
as gelatine and glycerin, or sucrose and acacia gum; and
mouthwashes comprising the active ingredient in a suitable liquid
carrier.
[0105] Compositions suitable for topical administration to the skin
may comprise the compounds dissolved or suspended in any suitable
carrier or base and may be in the form of lotions, gel, creams,
pastes, ointments and the like. Suitable carriers include mineral
oil, propylene glycol, polyoxyethylene, polyoxypropylene,
emulsifying wax, sorbitan monostearate, polysorbate 60, cetyl
esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and
water. Transdermal patches may also be used to administer the
compounds of the invention.
[0106] Compositions for rectal administration may be presented as a
suppository with a suitable base comprising, for example, cocoa
butter, glycerin, gelatine or polyethylene glycol.
[0107] Compositions 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.
[0108] Compositions suitable for parenteral administration include
aqueous and non-aqueous isotonic sterile injection solutions which
may contain anti-oxidants, buffers, bactericides and solutes which
render the composition isotonic with the blood of the intended
recipient; and aqueous and non-aqueous sterile suspensions which
may include suspending agents and thickening agents. The
compositions may be presented in unit-dose or multi-dose sealed
containers, for example, ampoules and vials, and may be stored in a
freeze-dried (lyophilised) condition requiring only the addition of
the sterile liquid carrier, for example water for injections,
immediately prior to use. Extemporaneous injection solutions and
suspensions may be prepared from sterile powders, granules and
tablets of the kind previously described.
[0109] Finally, formulations of these compositions in dry powder
form for delivery by a dry powder inhaler offer yet another means
of administration. This overcomes many of the disadvantages of the
oral and intravenous routes.
[0110] Preferred unit dosage compositions are those containing a
daily dose or unit, daily sub-dose, as herein above described, or
an appropriate fraction thereof, of the active ingredient.
[0111] It should be understood that in addition to the active
ingredients particularly mentioned above, the compositions may
include other agents conventional in the art having regard to the
type of composition in question, for example, those suitable for
oral administration may include such further agents as binders,
sweeteners, thickeners, flavouring agents disintegrating agents,
coating agents, preservatives, lubricants and/or time delay agents.
Suitable sweeteners include sucrose, lactose, glucose, aspartame or
saccharine. Suitable disintegrating agents include cornstarch,
methylcellulose, polyvinylpyrrolidone, xanthan gum, bentonite,
alginic acid or agar. Suitable flavouring agents include peppermint
oil, oil of wintergreen, cherry, orange or raspberry flavouring.
Suitable coating agents include polymers or copolymers of acrylic
acid and/or methacrylic acid and/or their esters, waxes, fatty
alcohols, zein, shellac or gluten. Suitable preservatives include
sodium benzoate, vitamin E, alpha-tocopherol, ascorbic acid, methyl
paraben, propyl paraben or sodium bisulphite. Suitable lubricants
include magnesium stearate, stearic acid, sodium oleate, sodium
chloride or talc. Suitable time delay agents include glyceryl
monostearate or glyceryl distearate.
[0112] In a further embodiment, the present invention provides
novel compounds, pharmaceutical compositions of said novel
compounds and therapeutic applications of said novel compounds for
the treatment of microbial infections.
[0113] Accordingly, in another aspect the present invention
provides a compound of the formula (I) or a salt thereof; ##STR5##
wherein:
[0114] A is O, S, CH.sub.2, CHF, CF.sub.2 or NR;
[0115] R.sup.1, R.sup.2, R.sup.2', R.sup.3, R.sup.3', and R.sup.4
are independently selected from the group consisting of H, halogen,
OH, N.sub.3, CN, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted aryl, optionally substituted acyl, optionally
substituted arylalkyl, optionally substituted heterocyclyl,
optionally substituted heteroaryl, optionally substituted
cycloalkyl, optionally substituted cycloalkenyl, optionally
substituted alkyloxy, optionally substituted alkenyloxy, optionally
substituted alkynoxy, optionally substituted aryloxy, optionally
substituted acyloxy, optionally substituted oxyacyl, optionally
substituted arylalkoxy, optionally substituted heterocycloxy,
optionally substituted heteroaryloxy, optionally substituted
cycloalkoxy, optionally substituted cycloalkenoxy, optionally
substituted amino, optionally substituted aminoacyl, optionally
substituted aminoacyloxy, optionally substituted acylamino,
optionally substituted oxyacylamino, optionally substituted
oxyacyloxy, optionally substituted acylimino, optionally
substituted acyliminoxy, optionally substituted oxyacylimino,
optionally substituted aminothioacyl, optionally substituted
thioacylamino, optionally substituted aminosulfinyl, optionally
substituted aminosulfonyl, optionally substituted thio, optionally
substituted thioacyl, optionally substituted thioacyloxy,
optionally substituted oxythioacyl, optionally substituted
oxythioacyloxy, optionally substituted phosphorylamino, optionally
substituted sulfinyl, optionally substituted sulfonyl, optionally
substituted sulfinylamino, optionally substituted sulfonylamino,
optionally substituted oxysulfinylamino, and optionally substituted
oxysulfonylamino, or R.sup.2 and R.sup.2' together or R.sup.3 and
R.sup.3' together represents .dbd.O, .dbd.S, or =L-Y' where L is N,
CH, CF, CCl or CBr and Y' is H, halogen, N.sub.3, methyl, ethyl or
CN;
[0116] R.sup.4' is --CY.sub.2SH, --CY.sub.2OH, --CY.sub.2NH.sub.2,
or -L'-R.sup.5;
[0117] L' is selected from the group consisting of --CY.sub.2--,
--CY.sub.2CY.sub.2--, --CY.sub.2OCY.sub.2--, --CY.sub.2SCY.sub.2--
and --CY.sub.2NHCY.sub.2--;
[0118] each Y is independently selected from the group consisting
of H, F, Cl, Br, OR, C.sub.1-C.sub.6alkyl, C.sub.2-C.sub.6alkenyl
and C.sub.2-C.sub.6alkynyl, wherein C.sub.1-C.sub.6alkyl,
C.sub.2-C.sub.6alkenyl, and C.sub.2-C.sub.6alkynyl may be
optionally substituted with one or more groups selected from F, Cl,
Br, OH, COOH, COOCH.sub.3, SH, SCH.sub.3, NH.sub.2, NHCH.sub.3,
N(CH.sub.3).sub.2, CN, NO.sub.2, C(O)NH.sub.2, C(O)NHCH.sub.3,
N.sub.3, C(S)NH.sub.2, OCH.sub.3, and OCH.sub.2CH.sub.3;
[0119] R.sup.5 is selected from the group consisting of OR,
NR.sub.2, monophosphate, diphosphate, and triphosphate, or a mono,
di or triphosphate mimic;
[0120] each R is independently selected from the group consisting
of H, CF.sub.3, optionally substituted alkyl, optionally
substituted alkenyl, optionally substituted alkynyl, optionally
substituted aryl, optionally substituted acyl, optionally
substituted cycloalkyl, optionally substituted cycloalkenyl,
optionally substituted heteroaryl, optionally substituted
heterocyclyl, and optionally substituted arylalkyl;
[0121] B is a group of formula (II) ##STR6##
[0122] wherein, if Z is a participant in a .pi. bond (double bond),
Z is independently selected from N or C-G; or, if Z is not a
participant in a .pi. bond (double bond), Z is independently
selected from O, S, Se, NR, NOR, NNR.sub.2, CO, CS, CNR, SO,
S(O).sub.2, SeO, Se(O).sub.2 or C(G).sub.2, wherein each G is
independently selected from the group consisting of H, halogen, OR,
SR, NR.sub.2, NROR, N.sub.3, COOR, CN, CONR.sub.2, C(S)NR.sub.2,
C(.dbd.NR)NR.sub.2, NCONR.sub.2, NCSNR.sub.2 and R; and
[0123] where any two adjacent Z are not both selected from O, S,
and Se, or not both selected from CO, CS, CNR, SO, S(O).sub.2, SeO,
and Se(O).sub.2;
[0124] wherein, if X is a participant in a .pi. bond (double bond),
X is C; or if X is not a participant in a .pi. bond (double bond),
X is CR or N;
[0125] wherein, if R'' is a participant in a .pi. bond (double
bond), R'' is O, S, Se, NR, NOR, and NNR.sub.2; or if R'' is not a
participant in a 71 bond (double bond), R'' is OR, SR, F, Cl, R, or
SeR;
[0126] dashed lines (---) indicate a possible .pi. or double bond;
and
[0127] wherein when R.sup.2', R.sup.3', and R.sup.5 are OH or
OC(O)CH.sub.3, L' is CH.sub.2, A is O, and R.sup.1, R.sup.2,
R.sup.3 and R.sup.4 are H, B is not the group of formula (III),
formula (IV), formula (V), formula (VI), or formula (VII)
##STR7##
[0128] (where all X* are H, or one of X* is CH.sub.3 and the other
two X are H);
[0129] when B is a group of formula (III), formula (VIII), or
formula (IX) ##STR8##
[0130] R.sup.2 and R.sup.2' are not both H;
[0131] when R.sup.2, R.sup.3' and R.sup.5 are OH, L' is CH.sub.2, A
is O and R.sup.1, R.sup.2', R.sup.3 and R.sup.4 are H, B is not the
group of formula (IV); and
[0132] when R.sup.2' is F, R.sup.3' is OH, R.sup.5 is triphosphate,
L' is CH.sub.2, A is O, and R.sup.1, R.sup.2, R.sup.3 and R.sup.4
are H, B is not a group of formula (X) ##STR9##
[0133] In some preferred embodiments of the invention one or more
of the following definitions apply:
[0134] The bicyclic base structure B of formula II may have one or
more ring double bonds and, in some instances, may have two or more
ring double bonds. Preferably, the base structure has at least two
double bonds and more preferably three or more double bonds.
[0135] Preferably the base structure B is selected form the
following formulae (XI) to (XXI) ##STR10## ##STR11## wherein Z, X
and R'' are as defined above.
[0136] Examples of preferred base structures (B) are as follows:
##STR12## ##STR13## ##STR14## wherein G and R are as defined
above.
[0137] Preferably X is N.
[0138] More preferably, the structure of formula II is represented
by the structure of formula IIa ##STR15## wherein each Z' is
independently N (if a participant in a .pi. bond) or NR (if not a
participant in a .pi. bond), and R'', R and Z are as defined
above.
[0139] Even more preferably, the structure of formula II is
represented by the structure of formula IIb ##STR16## wherein R''
and Z' are as defined above and each Z is independently CG (if a
participant is .pi. bond) or >C(G).sub.2 (if not a participant
in a .pi. bond).
[0140] Most preferably, the structure of formula II is represented
by the structures of formulae IIc, IId and IIe ##STR17## wherein R
and G are as defined above.
[0141] Preferably, for the compounds of formulae IIc, IId and IIe
each R is independently selected from H, optionally substituted
alkyl, optionally substituted aryl, optionally substituted
heteroaryl, optionally substituted heterocyclyl, and optionally
substituted arylalkyl and each G is independently selected from H,
halogen, CF.sub.3, optionally substituted alkyl, optionally
substituted aryl, optionally substituted heteroaryl, optionally
substituted heterocyclyl, and optionally substituted arylalkyl.
[0142] In a further preferred embodiment L' is --CH.sub.2--.
[0143] In yet a further preferred embodiment R.sup.4' is selected
from --CH.sub.2--OH, --CF.sub.2OH, --CCl.sub.2--OH,
--C(CH.sub.3)(CH).sub.3OH, --CH(CH.sub.3)OH,
--CH.sub.2--CH.sub.2--P(O)(OH).sub.2,
--CH.sub.2--CH.sub.2--P(O)(OH).sub.2, --CH.sub.2SP(O)(OH).sub.2,
--CH.sub.2SH, --CF.sub.2SH, and
--CH.sub.2--O--P(O)(OPh)(NHCH(CH.sub.3)(CO.sub.2Me)).
[0144] More preferably R.sup.4' is --CH.sub.2--OH.
[0145] Preferably, R.sup.1, R.sup.2, R.sup.2', R.sup.3, R.sup.3',
and R.sup.4 are independently selected from the group consisting of
H, halogen (more preferably F), OH, N.sub.3, CN, optionally
substituted alkyl, optionally substituted alkenyl, optionally
substituted alkynyl, optionally substituted aryl, optionally
substituted acyl, optionally substituted arylalkyl, optionally
substituted heterocyclyl, optionally substituted heteroaryl,
optionally substituted alkyloxy, optionally substituted acyloxy,
optionally substituted oxyacyl, optionally substituted amino,
optionally substituted aminoacyl, optionally substituted
aminoacyloxy, optionally substituted acylamino, optionally
substituted thio, or R.sup.2 and R.sup.2' together or R.sup.3 and
R.sup.3' together represents .dbd.O, .dbd.S, or =L-Y' where L is N,
CH, CF, CCl or CBr and Y' is H, halogen, N.sub.3, methyl, ethyl or
CN.
[0146] Even more preferably R.sup.1, R.sup.2, R.sup.2', R.sup.3,
R.sup.3', and R.sup.4 are independently selected from the group
consisting of H, F, Cl, Br, I, OH, SH, NH.sub.2, NHOH, NHNH.sub.2,
N.sub.3, COOH, CN, CONH.sub.2, C(S)NH.sub.2, COOR, R, OR, SR, SSR,
NHR, and NR.sub.2 wherein at least one of R.sup.2 or R.sup.2' is a
substituent other than H, and wherein R is selected from the group
consisting of optionally substituted alkyl, optionally substituted
alkenyl, optionally substituted alkynyl, optionally substituted
aryl, optionally substituted acyl, and optionally substituted
arylalkyl.
[0147] In yet another preferred embodiment, the sugar moiety is
selected from the following formulae: ##STR18## or C-5
monophosphate, diphosphate and triphosphate derivatives thereof, or
C-5 mono, di or triphosphate mimics.
[0148] In another preferred embodiment, at least one of R.sup.2 and
R.sup.2' is methyl, hydroxyl or F.
[0149] More preferably, the sugar moiety is selected from the
following formulae: ##STR19## or C-5 monophosphate, diphosphate and
triphosphate derivatives thereof, or C-5 mono, di or triphosphate
mimics.
[0150] Accordingly, in a preferred embodiment the compound of
formula (I) is selected from the following formulae, or salts
thereof: ##STR20## ##STR21## ##STR22## wherein:
[0151] each R on the sugar moiety is independently selected from H,
optionally substituted alkyl, optionally substituted alkenyl,
optionally substituted alkynyl, optionally substituted aryl,
optionally substituted acyl and optionally substituted
arylalkyl;
[0152] each R on the base moiety is independently selected from H,
optionally substituted alkyl, optionally substituted aryl,
optionally substituted heteroaryl, optionally substituted
heterocyclyl, and optionally substituted arylalkyl;
[0153] each G is independently selected from H, halogen, CF.sub.3,
optionally substituted alkyl, optionally substituted aryl,
optionally substituted heteroaryl, optionally substituted
heterocyclyl, and optionally substituted arylalkyl; and
[0154] C-5 monophosphate, diphosphate and triphosphate derivatives
thereof, or C-5 mono, di or triphosphate mimics.
[0155] While nucleosides incorporating a variety of sugar moieties
have been found to be useful for the inhibition of viral
polymerases, in the case of the Flaviviridae, and in particular,
the Hepatitis C virus, 2'-C-methyl ribnucleosides have been found
to be particularly useful (see Eldrup, A. B. et al., J. Med. Chem.
2004, 47(21), 5284-97, which is incorporated herein by
reference).
[0156] Accordingly, even more preferably, the sugar moiety is
represented by the following formula: ##STR23## or C-5
monophosphate, diphosphate and triphosphate derivatives thereof, or
C-5 mono, di or triphosphate mimics.
[0157] Most preferably, the sugar moiety is represented by the
following formula: ##STR24## or C-5 monophosphate, diphosphate and
triphosphate derivatives thereof, or C-5 mono, di or triphosphate
mimics.
[0158] The bicyclic nucleosides of the present invention also
include derivatives such as nucleotides, and nucleotide mimics
and/or prodrugs thereof.
[0159] In some embodiments, nucleotide mimics of the compounds of
the present invention of formula (I) discussed above include a
compound in which R.sup.5 is a monophosphate or monophosphate mimic
of formula (XXII) or (XXIII): ##STR25## where X.sup.1'X.sup.4', and
X.sup.6', independently are O, S, NR; X.sup.2', X.sup.3', and
X.sup.5' are selected independently from the group consisting of H,
F, NROR, N.sub.3, CN, (BH.sub.2G).sup.-M.sup.+,
(BH.sub.3).sup.-M.sup.+, R, OR, SR, and NR.sub.2. The substituents
(BH.sub.2G).sup.-M.sup.+ and (BH.sub.3).sup.-M.sup.+ are ion pairs,
which are linked to phosphorus through the negatively charged
boron. M.sup.+ is a cation, preferably a pharmaceutically
acceptable cation such as Ca.sup.2+, ammonium, trialkylammonium or
tertaalkylammonium, e.g., NH.sub.4.sup.+, Et.sub.3NH.sup.+,
Bu.sub.3NH.sup.+, and Bu.sub.4N.sup.+.
[0160] In some embodiments, nucleotide mimics of the compounds of
formula (I) as discussed above include di- and triphosphates and
di- and tri-phosphate mimics including a compound in which R.sup.5
is a di- or tri-phosphate moiety of formula (XXIV): ##STR26##
[0161] X.sup.2, X.sup.3, and X.sup.4 are selected independently
from the group consisting of O, S, Se, and NR;
[0162] X.sup.5 and X.sup.6 are selected independently from the
group consisting of --O--, --S--, --Se--, --CY.sub.2C(O)--,
--CH(OH)--, --C(OH).sub.2--, --CH.sub.2O--, --CH.sub.2CH.sub.2--,
--CH.sub.2CH(NH.sub.2)--, --CH.sub.2CH.sub.2CH(NH.sub.2)--,
--CY.sub.2OCY.sub.2--, --CY.sub.2--, --CRY--, --CY.sub.2CY.sub.2--,
--CHR--, --C.ident.C--, --HC.dbd.CH--, --NH--, --NR--, >NOH,
>NOR, >NNH.sub.2, and >NNHR;
[0163] X.sup.7, X.sup.8, X.sup.9, and X.sup.10 are selected
independently from the group consisting of H, F, OR, SR, NR.sub.2,
NROR, NRNR.sub.2, CN, N.sub.3, (BH.sub.3).sup.-M.sup.+,
(BH.sub.2G).sup.-M.sup.+, R and SeR;
[0164] Y, R, (BH.sub.2G).sup.-M.sup.+ and (BH.sub.3).sup.-M.sup.+
are as defined above; and
[0165] n is 0 or 1.
[0166] Additional nucleotide phosphate mimics and methods of making
the phosphate mimics appropriate for the compounds of the present
invention are described, inter alia, in WO 2003/072757 and WO
2003/073989, the entire contents of which are incorporated herein
by reference. Many of the nucleotide mimics discussed herein can be
prepared by similar approaches as published or by using well-known
knowledge of organophosphorous chemistry. Generally, phosphate
mimics of the nucleosides and nucleotides of the present invention
can inhibit enzyme function without phosphorylation and/or have
enhanced nuclease stability relative to nucleotides with unmodified
phosphate.
[0167] Accordingly, the term "phosphate mimic", unless otherwise
specified, refers to a phosphate analog, including, but not limited
to, phosphonate, phosphothiolate, phosphoselenoate,
selenophosphate, thiophosphate, P-boranophosphate, phosphoramidate,
sulfamate, sulfonate, and sulfonamide and/or a combination thereof.
Preferred embodiments of the phosphate mimics include phosphonate,
phosphoramidate, phosphorothioate, methylphosphonate,
fluoromethylphosphonate, difluoromethylphosphonate,
vinylphosphonate, phenylphosphonate, sulfonate, fluorophosphate,
dithiophosphorothioate, 5'-methylenephosphonate,
5'-difluoromethylenephosphonate, 5'-deoxyphosphonate,
5'-aminophosphoramidate, and 5'-thiophosphate. More preferred is
phosphonate and phosphoramidate.
[0168] Also, it will be appreciated that the term "diphosphate
mimic" and "triphosphate mimic" specifically refer to a diphosphate
analog and a triphosphate analog, respectively, which comprises at
least one of the phosphate mimics, one of the modifications at the
bridging site of diphosphate and triphosphate (e.g., X.sup.5,
X.sup.6 and X.sup.10), and/or replacements of non-bridging
phosphate oxygens (e.g., X.sup.4, X.sup.3 and X.sup.2).
[0169] The .alpha.-P, .beta.-P, and .gamma.-P in the mono, di- and
triphosphate mimics may independently adopt either R or S
configurations when chiral.
[0170] Accordingly, in compounds of formula (I) where a chiral
centre is present, the invention encompasses enantiomers, or
stereoisomers and mixtures thereof, such as enantiomerically
enriched mixtures. It will also be appreciated that the base
moieties of the present invention may exist as rapidly
interconvertible mixtures of isomers. Isomerism of this kind is
known in the art as tautomerism. Individual isomers are called
tautomers. Where tautomerism is possible the present invention
covers all possible tautomers of the compounds of formula (I).
[0171] The compounds of the present invention can be administered
to a subject as a pharmaceutically acceptable salt thereof. It will
be appreciated however that non-pharmaceutically acceptable salts
also fall within the scope of the present invention since these may
be useful as intermediates in the preparation of pharmaceutically
acceptable salts.
[0172] Suitable pharmaceutically acceptable salts include, but are
not limited to salts of pharmaceutically acceptable inorganic acids
such as hydrochloric, sulphuric, phosphoric, nitric, carbonic,
boric, sulfamic, and hydrobromic acids, or salts of
pharmaceutically acceptable organic acids such as acetic,
propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric,
maleic, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic,
phenylacetic, methanesulphonic, toluenesulphonic,
benezenesulphonic, salicyclic sulphanilic, aspartic, glutamic,
edetic, stearic, palmitic, oleic, lauric, pantothenic, tannic,
ascorbic and valeric acids.
[0173] Base salts include, but are not limited to, those formed
with pharmaceutically acceptable cations, such as sodium,
potassium, lithium, calcium, magnesium, ammonium and alkylammonium.
In particular, the present invention includes within its scope
cationic salts eg sodium or potassium salts, or alkyl esters (eg
methyl, ethyl) of the phosphate group.
[0174] Basic nitrogen-containing groups may be quarternised with
such agents as lower alkyl halide, such as methyl, ethyl, propyl,
and butyl chlorides, bromides and iodides; dialkyl sulfates like
dimethyl and diethyl sulfate; and others.
[0175] In some embodiments, the bicyclic nucleosides and
nucleotides of the present invention also include their prodrug
derivatives. The term "prodrug" is used in its broadest sense and
encompasses those derivatives that are converted in vivo to the
compounds of the invention. Prodrugs of the compounds of the
present invention may be prepared by modification of the sugar
moiety or of the phosphate or phosphate mimic to include a prodrug
substituent. Such prodrug modification is generally performed to
enhance drug absorption and/or drug delivery into cells.
[0176] Prodrugs substituents include, but are not limited to
residues of: proteins; antibiotics; D- and L-amino acids which may
be attached to a phosphate moiety or a phosphate mimic moiety via a
carbon atom (phosphonates), a nitrogen atom (phosphoamidates), or
an oxygen atom (phosphoesters) or may be attached to the sugar
moiety through any one or more of the R.sup.1-R.sup.5 groups;
peptides (preferably up to 10 amino acids) attached to a phosphate
moiety or a phosphate mimic moiety via a carbon atom
(phosphonates), a nitrogen atom (phosphoamidates), or an oxygen
atom (phosphoesters), or may be attached to the sugar moiety
through any one or more of the R.sup.1-R.sup.5 groups; drug
moieties attached to a phosphate moiety or a phosphate mimic moiety
via a carbon atom (phosphonates), a nitrogen atom
(phosphoamidates), or an oxygen atom (phosphoesters), or may be
attached to the sugar moiety through any one or more of the
R.sup.1-R.sup.5 groups; as well as including steroids; vitamins;
polyamines; carbohydrates; polyethylene glycols (PEGs);
cyclosaligenyls; substituted 4 to 8-membered rings, with or without
heteroatom substitutions, 1,3-phosphoamidate attachments to a
terminal phosphate or phosphate mimic moiety (.gamma. or .beta.) or
connecting between an .alpha.,.beta. or .beta.,.gamma. of a
phosphate moiety or phosphate mimic moiety, and so on.
[0177] In addition to those described herein, prodrug derivatives
of nucleosides, nucleotides and nucleotide phosphate mimics and
methods of making the prodrugs appropriate for use in the present
invention are described, inter alia, in PCT Publications WO
2003/072757 and WO 2003/073989.
[0178] The prodrug of a nucleoside 5'-monophosphate mimic can mask
the negative charges of the phosphate mimic moiety entirely or
partially, or mask the negative charges of the di-phosphate mimic
or tri-phosphate mimic moiety or phosphate moiety in order to
enhance drug absorption and/or drug delivery into cells.
[0179] In one embodiment a combination of prodrug substituents may
be attached to one or more X.sup.2', X.sup.3' and X.sup.5'
positions on a nucleoside mono-phosphate mimic or to one or more
X.sup.7-X.sup.10 positions on a nucleoside di- or tri-phosphate
mimic. Preferred prodrug substituents in positions X.sup.2',
X.sup.3' or X.sup.5' position include 2,3-O-diacylglyceryloxy,
2,3-O-dialkylglyceryloxy, 1-O-alkyl-2-O-acylglyceryloxy,
1-O-acyl-2-O-alkylglyceryloxy,
1-S-alkyl-2-O-acyl-1-thioglyceryloxy, acyloxymethoxy,
S-acyl-2-thioethoxy, S-pivaloyl-2-thioethoxy, acyloxymethoxy,
pivaloyloxymethoxy, alkoxycarbonyloxymethoxy,
S-alkyldithio-S'-ethyoxy acyloxymethoxy, S-acyl-2-thioethoxy,
S-pivaloyl-2-thioethoxy, pivaloyloxymethoxy,
alkoxycarbonyloxymethoxy, and S-alkyldithio-S'-ethyoxy.
[0180] In a further embodiment, the prodrug substituent is a
substituent on a hydroxyl group of the sugar moiety (that is, for
instance, any one of R.sup.1-R.sup.5). Preferably, the modification
results in the formation of an ester and in this regard the
preferred prodrug substituents are C.sub.1-C.sub.6acyl groups for
example, acetyl, propionyl, pivaloyl, etc. Also preferred are
substituted C.sub.1-C.sub.6 acyl moieties, for example,
fluoroacetyl, difluoroacetyl, etc. More preferably the substituted
C.sub.1-C.sub.6 acyl group is represented as a residue of an L or D
amino acid consisting of alanine, asparagine, cysteine, glutamine,
glycine, isoleucine, leucine, methionine, phenylalanine, proline,
serine, threonine, tryptophan, tyrosine, valine, aspartic acid,
glutamic acid, arginine, histidine, and lysine. Most preferably the
prodrug substituent is an amino acid residue of D or L-valine.
[0181] The prodrug can be activated either by cellular enzymes such
as lipases, esterases, reductases, oxidases, nucleases or by
chemical cleavage such as hydrolysis to release (liberate) either
the nucleoside, nucleotide or nucleotide mimic after the prodrug
enters cells.
[0182] In addition to using prodrug approaches, the delivery of the
nucleosides and nucleotides may be assisted by using a
therapeutically acceptable carrier such as liposomal suspensions,
cationic lipids, and polyimines.
[0183] The novel nucleosides, nucleotides, nucleotide mimics and
prodrugs thereof, of the present invention can be prepared by those
who are skilful in synthetic organic and nucleoside chemistry using
established synthetic methodology (Chemistry of Nucleosides and
Nucleotides Vol. 1 (1988), Vol. 2 (1991), Vol. 3 (1994), edited by
L. B. Townsend, Plenum Press; Handbook of Nucleoside Synthesis by
H. Vorbruggen and C. Ruh-Pohlenz, John Wiley & Sons, Inc.,
2001; The Organic Chemistry of Nucleic Acids by Y. Mizuno,
Elsevier, 1986).
[0184] The nucleosides of the present invention can be converted to
their corresponding monophosphate, diphosphate, and triphosphate
nucleosides by established phosphorylation procedures. Similarly,
known methods in the art can be used to synthesise the nucleotide
and phosphate mimics and prodrugs. The following schemes and
description serve as representative syntheses of the nucleosides
and nucleotides of the present invention. As such, other compounds
such as those having --CY.sub.2SH, --CY.sub.2OH or -L'-R.sup.5
groups other than CH.sub.2R.sup.5 may similarly be made.
[0185] The bicyclic nucleosides of the present invention may be
prepared by modification of optionally protected and functionalised
cytosine, uracil and other base analogues followed by Stille, Heck,
Sonogashira or other metal-mediated cross coupling chemistry to
introduce an .alpha.,.beta.-unsaturated ester, alkyne or other
functional group. Such processes allow for stereoselective
synthesis of an intermediate capable of efficient cyclisation to
form the bicyclic compounds of the present invention. Cyclisation
and optional deprotection of the product delivers the target
bicyclic nucleoside.
[0186] Any compound capable of metal-mediated cross coupling may be
used, such as a tin derivative like trialkyltin. More preferably
tributyltin. Preferably the reactions are carried out using a
palladium based coupling agent. Suitable coupling agents are known
in the art and include Pd(PPh.sub.3).sub.2Cl.sub.2,
Pd(PPh.sub.3).sub.4, Pd(dibenzylideneacetone), and
PdCl.sub.2(CH.sub.3CN).sub.2. Also, preferably the palladium
catalysed coupling reactions may also include a co-catalyst, for
instance, CuI which may be in the presence of a suitable
non-nucleophilic base such as a trialkylamine.
[0187] Coupling reactions are generally performed at temperatures
around room temperature. Elevated temperatures such as temperatures
between 30-80.degree. C. can be employed to effect coupling and
cyclisation in a single step. It is also preferred that such
reactions are carried out under an inert atmosphere of either
nitrogen or argon. Suitable solvents include ether solvents such as
THF and diethylether or polar solvents such as DMF.
[0188] For example, Schemes 1 and 2 illustrates some preferred
cyclisation methods for forming the 6-membered ring portions of the
bicyclic bases (B) of the compounds of the present invention.
##STR27##
[0189] Methodologies other than metal-mediated cross-coupling and
cyclisation can also be used to prepare the bicyclic nucleosides.
For example, the final process in scheme 2 depicts how a
bifunctional two atom unit (such as an .alpha.-halocarboxylic ester
or glyoxal) can be used to form the second 6-membered ring portion
of a suitable nucleoside intermediate. ##STR28##
[0190] In the cyclisation methodology depicted in Schemes 1 and 2
each of Z.sup.1-Z.sup.4 is independently Z.
[0191] Preferably, in this process A is O, CH.sub.2 or optionally
protected N; Y is halogen or other appropriate group such as
trifluoromethanesulfonate; W.sup.4 is H or trialkyltin; Z.sup.1 and
Z.sup.2 are each independently C, CH, C-halogen, C-alkyl, C-aryl,
C-heteroaryl, C--O-alkyl or C--S-alkyl; Z.sup.3 is CH, C-alkyl,
C-halogen, N, CNHR, CNH.sub.2, CNR.sub.2, C.dbd.O, or C.dbd.S;
Z.sup.4 is CH, C-halogen, C-alkyl, C-aryl, C-heteroaryl,
C--O-alkyl, C--S-alkyl, C--OH, C--NH.sub.2, C--NHR, C--NR.sub.2 or
C.dbd.O; R.sup.1, R.sup.2, R.sup.2'R.sup.3, R.sup.3', R.sup.4' are
each independently H, halogen, alkyl, O-alkyl, OH, optionally
protected O, methyl, H or F; and R.sup.5 is an optionally protected
OH or NH.sub.2; and (--) denotes an optional double bond.
[0192] The bicyclic bases of the present invention can be further
modified either prior to being added to the sugar moiety or once
attached. Such modification may involve transformations through
functionalisation, defunctionalisation, or functional group
interconversion. Modifications may include esterification, the
preparation of enol ethers, o-alkylation, bromination,
hydrogenation, dihydroxylation, epoxidation, oximation, and
amination.
[0193] The compounds described herein can also be converted into
their corresponding mono-, di- and triphosphates using well
established methods. Furthermore, as discussed above prodrugs of
mono-, di- and triphosphates can be prepared in order to optimise
the biological efficacy of these phosphorylated compounds. Methods
for preparing such prodrugs are well known in the art (see Wagner,
C. R., et al. Med. Res. Rev., 2000, 20, 417-451). ##STR29##
[0194] In Scheme 3, preferably A is O, CH.sub.2 or optionally
protected N; R.sup.1, R.sup.2, R.sup.2', R.sup.3, R.sup.3', R.sup.4
are each independently H, halogen, alkyl, O-alkyl, OH, optionally
protected O, or methyl and B is as described herein.
[0195] As discussed earlier an alternative to the use of phosphates
is the use of phosphate mimics and their prodrugs. One such
phosphate mimic is shown below and this can be prepared using
appropriately protected nucleosides and known conditions.
##STR30##
[0196] In Scheme 4, preferably A is O, CH.sub.2 or optionally
protected N; R.sup.1, R.sup.2, R.sup.2', R.sup.3, R.sup.3', R.sup.4
are each independently H, halogen, alkyl, O-alkyl, OH, optionally
protected O, methyl or F; X' is O, S, NH, CF.sub.2, CHF, CClH,
CBr.sub.2 or CHBr; B is as described herein.
[0197] The bicyclic compounds of the present invention may be
tested for biological activity using well known procedures.
[0198] Antiviral assays are conducted according to published,
widely used protocols. In order to obtain the therapeutic index,
compound-induced cytotoxicity to host cells is also measured in
parallel with antiviral activities. To determine the mode of action
of antiviral nucleosides the corresponding nucleoside triphosphates
are subject to enzyme-based assays for the inhibition of viral
polymerases according to known protocols (Ranjith-Kumar et al. J.
Virol. 2001, 75, 8615; Dhanak et al. J. Biol. Chem. 2002, 277,
38322-38327). Some compounds of the present invention showed
K.sub.i values of less than 1 .mu.M against HCV NS5B.
[0199] Since the replicon RNA replication mimics the replication of
HCV RNA in infected hepatocytes, compounds that have the inhibitory
effects in replicon assays are potentially useful as anti-HCV
drugs. The HCV replicon-containing cell lines (Randall and Rice,
Current Opinion in Infectious Diseases 2001, 14, 743) are used for
the identification of potential anti-HCV compounds. Among them is a
widely used subgenomic replicon system developed by Lohmann et al.
(Science 1999, 285, 110; J. General Virol. 2000, 81, 1631; J.
Virol. 2001, 75, 1437, 2002, 76, 4008). Some compounds of the
present invention showed potent anti-HCV activity with EC.sub.50
values of low .mu.M.
[0200] Widely used protocols developed by Korba et al. (Antiviral
Res. 1992, 19, 55), and Pai et al. (Antimicrobial Agents Chemother.
1996, 40, 380) are useful for the determination of in vitro
anti-HBV activity.
[0201] Anti-HIV assays can be conducted according to the protocols
developed by Schinazi et al. (Antimiromobial Agents Chemother.
1990, 34, 1061; 1992, 36, 2423; 1993, 37, 875) or other widely used
protocols (Kimpton et al J. Virol. 1992, 66, 2232; Chan et al. J.
Med. Chem. 2001, 44, 1866).
[0202] Preferred nucleoside triphosphates of the present invention
may act as potent inhibitors of the non-structural position SB
(NS5B) which is HCV's RNA-dependent RNA polymerase. Accordingly,
such compounds are preferably suited to treat and/or prevent HCV.
Also, as the preferred novel compounds of the present invention are
expected to exhibit novel profiles of activity they may provide the
artisan with an alternative to treating viruses which display drug
resistance to conventional drugs. Other advantages which may be
exhibited by the preferred novel compounds of the present invention
include: [0203] reduced toxicity and tolerability relative to
existing therapies and those in development; and/or [0204] improved
pharmacokinetic properties.
[0205] Accordingly, nucleosides, nucleotide, nucleotide mimics
and/or their prodrugs of the present invention may be useful for
the inhibition of a variety of enzymes including, but not limited
to, DNA or RNA polymerases, helicases, ribonucleotide reductases,
protein kinases, and telomerases and for the modulation of
G-proteins, P2 purinergic receptors and the allosteric sites of a
variety of enzymes. Preferably, the novel nucleosides, nucleotides,
nucleotide mimics and/or prodrugs of the present invention are used
to treat viral infections caused by the RNV viruses of the group
Flaviviridae and, in particular, HCV.
[0206] The novel nucleosides, nucleotides, nucleotide mimics and/or
their prodrugs of the present invention are useful for the
treatment of infectious diseases caused by infectious agents such
as parasites, bacteria and fungi.
[0207] Also, the novel nucleosides, nucleotide mimics and/or their
prodrugs that display potent cytotoxicities to fast-dividing
cancerous cells may be useful for the treatment of proliferative
disorders, including, but not limited to, lung cancer, liver
cancer, prostate cancer, colon cancer, breast cancer, ovarian
cancer, melanoma, and leukemia.
[0208] As the ligands of P2 receptors and G-proteins as well as the
inhibitors of protein kinases, the novel nucleosides, nucleotides,
nucleotide mimics and/or their prodrugs of the present invention
may also be useful for the treatment of a wide range of other
diseases and disorders such as inflammatory diseases, autoimmune
diseases, Type 2 diabetes, and cardiovascular diseases.
[0209] In order to overcome drug resistance, combination therapies
are widely used in the treatment of infectious diseases and
proliferative disorders. The nucleosides, nucleotides, nucleotide
mimics and/or their prodrugs of the present invention may be
therapeutically administered as a single drug, or alternatively may
be administered in combination with one or more other active
chemical entities to form a combination therapy. The other active
chemical entities may be a small molecule, a polypeptide, or a
polynucleotide.
[0210] For instance, compounds of this invention may be
particularly useful when used in combination with other agents
known to exert an antiviral effect. For example, combination with
immunomodulatory/antiviral agents such as interferons, interferon
derivatives and other large or small molecules known to modulate
host immune responses may be beneficial. Similarly, combinations of
compounds of this invention with IMPDH inhibitors (e.g.,
ribavirin), antiviral nucleosides, antiviral non-nucleosides (e.g.,
polymerase inhibitors, protease inhibitors) could augment the
activity of the bicyclic nucleosides when administered alone.
[0211] The following examples are put forth so as to provide those
of ordinary skill in the art with a complete disclosure and
description of how to prepare and use the compounds disclosed and
claimed herein.
[0212] All references mentioned herein are incorporated herein by
reference in their entirety.
[0213] The abbreviations that may be used herein, including the
Schemes and experimental section are as follows unless indicated
otherwise:
[0214] Bu: n-butyl
[0215] Bn benzyl
[0216] Bz: benzoyl
[0217] DCM: dichloromethane
[0218] DIEA: diisopropylethylamine
[0219] DMF: dimethylformamide
[0220] Et: ethyl
[0221] EtOAc: ethyl acetate
[0222] Me: methyl
[0223] MeOH: methyl alcohol
[0224] MS: mass spectrometry
[0225] NMR: nuclear magnetic resonance
[0226] Ph: phenyl
[0227] HPLC: high performance liquid chromatography
[0228] TEA: triethylamine
[0229] TFA: trifluoroacetic acid
[0230] THF: tetrahydrofuran
[0231] The following Examples are offered to illustrate but not to
limit the invention.
EXAMPLES
Synthesis
[0232] TABLE-US-00002 TABLE 1 Example Structure Molec Formula MWt 1
##STR31## C.sub.13H.sub.15N.sub.3O.sub.6 309.28 2 ##STR32##
C.sub.13H.sub.14IN.sub.3O.sub.6 435.17 3 ##STR33##
C.sub.13H.sub.14BrN.sub.3O.sub.6 388.17 4 ##STR34##
C.sub.13H.sub.14ClN.sub.3O.sub.6 343.72 5 ##STR35##
C.sub.13H.sub.15N.sub.3O.sub.6 309.28 6 ##STR36##
C.sub.14H.sub.17N.sub.3O.sub.6 323.30 7 ##STR37##
C.sub.20H.sub.21N.sub.3O.sub.6 399.40 8 ##STR38##
C.sub.16H.sub.21N.sub.3O.sub.6 351.36 9 ##STR39##
C.sub.14H.sub.19N.sub.3O.sub.6 325.32 10 ##STR40##
C.sub.20H.sub.23N.sub.3O.sub.6 401.42 11 ##STR41##
C.sub.17H.sub.24N.sub.4O.sub.5 364.40 12 ##STR42##
C.sub.17H.sub.24N.sub.4O.sub.6 380.40 13 ##STR43##
C.sub.18H.sub.26N.sub.4O.sub.5 378.43 14 ##STR44##
C.sub.17H.sub.25N.sub.5O.sub.5 379.41 15 ##STR45##
C.sub.13H.sub.18N.sub.3O.sub.15P.sub.3 549.21
Experimental Data
[0233] .sup.1H and .sup.31P NMR spectra were recorded on either a
Bruker Avance DRX 400, AC 200 or AM 300 spectrometer. Spectra were
recorded in CDCl.sub.3, d.sub.6-acetone, CD.sub.3OD or d.sub.6-DMSO
using the residual solvent peak as a reference. Chemical shifts are
reported on the .delta. scale in parts per million (ppm) using the
following conventions to assign the multiplicity: s (singlet), d
(doublet), t (triplet), q (quartet) m (multiplet) and prefixed b
(broad). Mass spectra (ESI) were recorded on a Finnigan LCQ
Advantage spectrometer. All microwave reactions were carried out in
a CEM Discover microwave reactor. Flash and radial chromatography
was performed on 40-63 .mu.m silica gel 60 (Merck No. 9385).
Preparative HPLC was carried out using a Gilson 322 pump with a
Gilson 215 liquid handler and a HP1100 PDA detector. HPLC systems
employed Phenomonex C8(2) columns using either acetonitrile or
acetonitrile containing 0.06% TFA in water or water containing 0.1%
TFA. Alternatively, a Phenomonex C18 column was used with
acetonitrile and aqueous 1M triethylammonium acetate (primarily for
phosphate nucleotides). ##STR46## Intermediate A
[0234] N,O-Bis(trimethylsilyl)acetamide (7.17 mL) was added to
5-iodocytidine (2.32 g) in dry acetonitrile (15 mL) and the mixture
was heated under argon at 80.degree. C. for 30 mins. A suspension
of commercially available
1,2,3,5-tetra-O-benzoyl-2-C-methyl-alpha/beta-D-ribofuranose (5.16
g) in dry acetonitrile (40 mL) was added and the mixture heated for
1 h. SnCl.sub.4 (1.71 mL) was added cautiously and heating
continued for 2 hrs. The reaction was cooled to ambient
temperature, poured into an ice cold solution of saturated aqueous
sodium bicarbonate and extracted with EtOAc (.times.3). The
combined organic extracts were washed with brine, dried and
evaporated in-vacuo. The residue was purified by flash
chromatography on silica eluting with 3% MeOH/DCM. The product was
obtained as a gum (3.36 g). MS m/z ([M+H].sup.+) 695.7;
([2M+H].sup.+) 1390.8.
Intermediate B
[0235] A degassed solution of intermediate A (1.39 g) and
(Z)-ethyl-3-(tributylstannyl) propenoate (1.56 g) was stirred in
dry DMF under argon for 10 mins. CuI (76 mg) and
PdCl.sub.2(PPh.sub.3).sub.2 (140 mg) were added and the mixture
heated at 70.degree. C. under argon for 16 hrs. The reaction was
cooled to ambient temperature, diluted with water (100 mL) and
filtered. The residue was washed with water, dissolved in EtOAc and
washed again with water. The organic solution was dried
(MgSO.sub.4) and the solvent evaporated in-vacuo to provide a brown
gum. The aqueous mother liquor was also extracted with EtOAc
(.times.2) and the combined organics dried (MgSO.sub.4), and
evaporated in-vacuo. The combined crude extracts were purified by
radial chromatography on silica eluting with 3%-5% MeOH/DCM to give
the required product as an oil, which, on trituration with ether
afforded a yellow crystalline solid (70 mg). MS m/z ([2M+H].sup.+)
1243.5. Further product was available from the mother-liquor.
Preparation of Intermediates C, D, E
[0236] Preparation of Intermediate C is an example of the general
method.
[0237] A mixture of Intermediate B (125 mg) and N-iodosuccinimide
(90.6 mg) in dry acetonitrile (1.50 mL) was heated in a microwave
reactor at 120.degree. C. for 30 mins (initial power 200 W). The
solution was evaporated in-vacuo, dissolved in EtOAc and washed
with 10% sodium metabisulphite, brine, and evaporated in-vacuo. The
crude material was purified by radial chromatography on silica
eluting with 3% MeOH/DCM to yield the required product (120 mg). MS
m/z ([2M+H].sup.+) 1494.3.
[0238] Similarly for Intermediates D and E.
Intermediate D
[0239] Intermediate B (62 mg) yielded intermediate D (45 mg). MS
m/z ([2M+H].sup.+) 1400.6, 1401.6, 1403.6.
Intermediate E
[0240] Intermediate B (62 mg) yielded 62 mg intermediate D. MS m/z
([2M+H].sup.+) 1310.6, 1311.7, 1313.7.
Preparation of Examples 1-4
[0241] Preparation of Example 1 is an example of the general
method.
Example 1
[0242] Intermediate B (60 mg) was suspended in dry methanol (1 mL)
under argon. A freshly prepared solution of 1M sodium methoxide in
methanol (0.5 mL) was added and the reaction stirred for 18 hrs.
Evaporation of the solvent in-vacuo with minimal heating and
purification by radial chromatography on silica eluting with 20%
MeOH/DCM yielded the required compound as a crystalline solid.
[0243] MS m/z ([M+H].sup.+) 309.9. 1H .sup.1H NMR
(CD.sub.3OD+D.sub.2O) .delta. 9.24 (s, 1H), 7.61 (d, 1H), 6.23 (d,
1H), 6.11 (s, 1H), 4.08-4.00 (m, 2H), 3.93-3.83 (m, 2H), 1.12 (s,
3H).
[0244] Similarly for Examples 2 and 3.
Example 2
[0245] Intermediate C (12 mg) yielded Example 2 (1.70 mg)
[0246] MS m/z ([M+H].sup.+) 435.7, ([2M+H].sup.+) 870.4. .sup.1H
NMR (CD.sub.3OD) .delta. 9.26 (s, 1H), 8.18 (s, 1H), 6.10 (s, 1H),
4.07-4.00 (m, 2H), 3.95-3.82 (m, 2H), 1.12 (s, 3H).
Example 3
[0247] Intermediate D (21 mg) yielded Example 3 (4.1 mg)
[0248] MS m/z ([M+H].sup.+) 387.7, 389.7. .sup.1H NMR (CD.sub.3OD)
.delta. 9.28 (s, 1H), 7.95 (s, 1H), 6.10 (s, 1H), 4.07-3.99 (m,
2H), 3.89-3.82 (m, 2H), 1.12 (s, 3H).
Example 4
[0249] Intermediate E (28 mg) was stirred with 7M NH.sub.3 in MeOH
at ambient temperature for 18 h. The reaction was evaporated to
dryness in-vacuo, dissolved in water (2 mL) and washed with DCM
(.times.3) and EtOAc. The aqueous phase was evaporated and the
residue triturated with ether to yield the required compound as a
yellow solid (12 mg).
[0250] MS m/z ([M+H].sup.+) 343.7, 345.7; ([2M+H].sup.+) 686.6,
688.6. .sup.1H NMR (CD.sub.3OD) .delta. 9.34 (s, 1H), 7.73 (s, 1H),
6.11 (s, 1H), 4.07-3.99 (m, 2H), 3.90-3.82 (m, 2H), 1.12 (s,
3H).
Example 5
[0251] ##STR47##
[0252] Example 5 (10.5 mg) was prepared directly from commercially
available 5-Iodo-2'-O-methylcytidine using the method described for
intermediate B in scheme 1.
[0253] MS m/z ([M+H].sup.+) 309.8; ([2M+H].sup.+) 618.6,
([2M+Na].sup.+) 640.8. .sup.1H NMR (CD.sub.3OD+D.sub.2O) .delta.
9.23 (s, 1H), 7.64 (d, 1H), 6.27 (d, 1H), 5.95 (s, 1H), 4.21-4.17
(m, 1H) 4.06-4.02 (m, 2H), 3.95-3.73 (m, 2H), 3.64 (s, 3H).
##STR48## ##STR49## Intermediate F
[0254] N,O-Bis(trimethylsilyl)acetamide (6.45 mL) and uracil (0.986
g) in dry acetonitrile (20 mL) were heated under argon at
80.degree. C. for 30 mins. A suspension of commercially available
1,2,3,5-tetra-O-benzoyl-2-C-methyl-alpha/beta-D-ribofuranose in dry
acetonitrile (80 mL) was added and heating continued for 1 hr.
SnCl.sub.4 was added cautiously and heating continued for 5 hrs.
The reaction was cooled to ambient temperature, poured into an ice
cold solution of saturated sodium bicarbonate (100 mL) and
extracted with EtOAc (.times.3). The combined organic extracts were
washed with brine, dried (MgSO.sub.4) and evaporated in-vacuo to
provide crude product as a frothy white gum (4.7 g) which was used
in subsequent reactions without further purification. MS m/z
([M+H].sup.+) 571, ([2M+Na].sup.+) 1163.
Intermediate G
[0255] 1,2,4-triazole (207 mg) was dissolved in dry acetonitrile (5
mL) under argon, cooled to 0.degree. C. and treated with POCl.sub.3
(75 .mu.L). After 5 mins, triethylamine (0.60 mL) was added and
stirring continued for 1 h at ambient temperature before addition
of Intermediate F in dry acetonitrile (5 mL). The reaction was
diluted with EtOAc after 90 mins and washed successively with
saturated NaHCO.sub.3 and brine (each .times.3). The organic layer
was dried (MgSO.sub.4) and evaporated in-vacuo to provide the crude
triazole intermediate as a yellow gum (127 mg). This gum was
suspended in MeOH (5 mL) and 2M MeNH.sub.2 in MeOH (0.20 mL) added
followed by vigorous stirring for 20 mins. After this time,
volatile materials were evaporated in-vacuo and the residue
purified by radial chromatography on silica eluting with 2%
MeOH/DCM to give the required product as a gum (46 mg). MS m/z
([M+H].sup.+) 583.9, ([2M+H].sup.+) 1166.9.
Intermediate H
[0256] The crude triazole intermediate (621 mg) was prepared using
the method described for intermediate G. This gum was suspended in
dry ethanol (10 mL) before addition of benzylamine (164 .mu.l).
After stirring for 1 hr at ambient temperature the solvent was
removed in-vacuo and the residue purified by radial chromatography
on silica eluting with 3%-5% MeOH/DCM to give the required product
as an oil (424 mg). MS m/z ([M+H].sup.+) 660.4, 661.4.
Intermediate I
[0257] The crude triazole intermediate (249 mg) was prepared using
the method described for intermediate G. This gum was suspended in
dry methanol (5 mL) before addition of N-propylamine (49 .mu.l).
After stirring for 30 mins at ambient temperature the solvent was
removed in-vacuo and the residue purified by radial chromatography
on silica eluting with 3% MeOH/DCM to give the required product as
gum (45 mg). MS m/z ([M+H].sup.+) 612.4, 613.4.
[0258] Intermediates J, K and L were prepared by the general method
described previously for Intermediate C.
Intermediate J
[0259] Intermediate G (200 mg) yielded Intermediate J (87 mg)
[0260] MS m/z ([M+H].sup.+) 709.7.
Intermediate K
[0261] Intermediate H (188 mg) yielded Intermediate K (110 mg).
[0262] MS m/z ([M+H].sup.+) 785.9.
Intermediate L
[0263] Intermediate I (130 mg) yielded Intermediate L (106 mg).
[0264] MS m/z ([M+H].sup.+) 738.2
[0265] Intermediates M, N and O were prepared by the general method
described previously for Intermediate B.
Intermediate M
[0266] A degassed solution of intermediate J (100 mg) and
(Z)-ethyl-3-(tributylstannyl) propenoate (110 mg) was stirred in
dry DMF under argon for 10 mins. CuI (5.5 mg) and
PdCl.sub.2(PPh.sub.3).sub.2 (10 mg) were added and the mixture
heated at 70.degree. C. under argon for 16 hr. The solvent was
evaporated in-vacuo and the residue partitioned between EtOAc and
water. The aqueous phase was further extracted with EtOAc
(.times.2) and the combined organic extracts dried (MgSO.sub.4) and
evaporated in-vacuo. The crude product was purified by radial
chromatography on silica eluting with 3% MeOH/DCM to provide a
yellow gum (56 mg). MS m/z ([M+H].sup.+) 635.8, ([2M+H].sup.+)
1270.7.
Intermediate N
[0267] Intermediate K (120 mg) yielded Intermediate N (19 mg).
[0268] MS m/z ([M+H].sup.+) 711.7, ([2M+H].sup.+) 1422.5.
Intermediate O
[0269] Intermediate L (105 mg) yielded Intermediate O (19 mg).
[0270] MS m/z ([M+H].sup.+) 663.9, ([2M+H].sup.+) 1326.9.
Example 6
[0271] Intermediate M (36 mg) was stirred with 7M NH.sub.3 in MeOH
(3 mL) at ambient temperature for 4 hrs. The reaction was
evaporated to dryness in-vacuo and the residue purified by radial
chromatography on silica eluting with DCM/MeOH/25% NH.sub.3 90:9:1
to yield the required compound as a yellow solid (6 mg).
[0272] MS m/z ([M+H].sup.+) 323.8. 1H .sup.1H NMR (CD.sub.3OD)
.delta. 9.28 (s, 1H), 7.51 (d, 1H), 6.33 (d, 1H), 6.12 (s, 1H),
4.08-3.99 (m, 2H), 3.91-3.86 (m, 2H), 3.53 (s, 3H), 1.12 (s,
3H).
[0273] Similarly for Examples 7 and 8.
Example 7
[0274] Intermediate N (19 mg) yielded Example 7 (3.9 mg).
[0275] MS m/z ([M+H].sup.+) 399.9. 1H .sup.1H NMR (CD.sub.3OD)
.delta. 9.30 (s, 1H), 7.54 (d, 10H), 7.40-7.35 (m, 2H), 7.26-7.17
(m, 3H), 6.36 (d, 1H), 6.12 (s, 1H), 5.44 (s, 1H), 4.07-3.99 (m,
2H), 3.91-3.81 (m, 2H), 1.12 (s, 3H).
Example 8
[0276] Intermediate O (38 mg) yielded Example 8 (8.0 mg).
[0277] MS m/z ([M+H].sup.+) 351.8. 1H .sup.1H NMR (CD.sub.3OD)
.delta. 9.30 (s, 1H), 7.50 (d, 1H), 6.32 (d, 1H), 6.12 (s, 1H),
4.21-4.16 (m, 2H), 4.08-3.99 (m, 2H), 3.91-3.83 (m, 2H), 1.74-1.61
(m, 2H), 1.12 (s, 3H), 0.93 (t, 3H).
Example 9
[0278] Example 6 (20 mg) was dissolved in methanol (3 mL) and
NH.sub.4Cl (10 mg) in water (0.2 mL) was added followed by
NaBH.sub.4 (10 mg). After 10 mins the reaction was evaporated to
dryness in-vacuo and the residue dissolved in water (3 mL) and
washed with DCM. The aqueous extract was then purified by
preparative HPLC to afford the product as an oil (4.23 mg).
[0279] MS m/z ([M+H].sup.+) 326.0. 1H .sup.1H NMR (CD.sub.3OD)
.delta. 7.26 (d, 1H), 6.10 (d, 1H), 5.84 (s, 1H), 4.60 (d, 1H),
3.98 (d, 1H), 3.92-3.68 (m, 4H), 3.50 (s, 3H), 1.22 (s, 3H).
[0280] Similarly for Example 10.
Example 10
[0281] Example 7 (20 mg) yielded Example 10 (7.0 mg).
[0282] MS m/z ([M+H].sup.+) 402.0. 1H .sup.1H NMR (CD.sub.3OD)
.delta. 7.28-7.18 (m, 6H), 6.08 (d, 1H), 5.75 (s, 1H), 5.45 (d,
1H), 5.38 (d, 1H), 4.62 (d, 1H), 4.01 (d, 1H), 3.93-3.89 (m, 1H),
3.80-3.68 (m, 3H), 1.19 (s, 3H). ##STR50## Intermediate P
[0283] A degassed solution of intermediate A (695 mg) and propargyl
alcohol (175 .mu.l) was stirred in dry DMF (7 mL) under argon. CuI
(38 mg) and PdCl.sub.2(PPh.sub.3).sub.2 (70 mg) were added followed
by TEA (278 .mu.l) and the mixture stirred at ambient temperature
for 18 hrs. The solvent was evaporated in-vacuo and the residue
partitioned between EtOAc and water. The organic layer was washed
with water, dried (MgSO.sub.4) and evaporated in-vacuo. The crude
product was purified by radial chromatography on silica eluting
with DCM then 5-10% MeOH/DCM to provide a gum (257 mg). MS m/z
([M+H].sup.+) 624.0, ([2M+H].sup.+) 1247.1.
Intermediate Q
[0284] Intermediate P (248 mg) was suspended in DCM (10 mL) and
treated with Dess-Martin periodinane (338 mg) at ambient
temperature. The reaction was diluted with DCM after 2 hrs and
washed with saturated aqueous NaHCO.sub.3. The organic phase was
evaporated in-vacuo and the residue purified by radial
chromatography on silica eluting with 5-10% MeOH/DCM to provide the
product as a gum (263 mg). MS m/z ([M+H].sup.+) 621.8,
([2M+H].sup.+) 1242.9.
Intermediate R
[0285] Intermediate Q (30 mg) and pyrrolidine (14 mg) in
acetonitrile (0.5 mL) were heated in a microwave reactor at
70.degree. C. for 10 mins (initial power 200 W). The solvent was
evaporated in-vacuo and the residue dissolved in EtOAc, washed with
water then 1M HCl. The organic layer was washed with water, dried
(MgSO.sub.4) and evaporated in-vacuo to yield crude cyclised
product (28 mg) used in the next step without further purification.
The material was dissolved in MeOH (2 mL) and NaBH.sub.3CN (13 mg)
was added. After 1 hr the solvent was evaporated in-vacuo and the
residue dissolved in EtOAc. The organic layer was washed
successively with saturated aqueous NaHCO.sub.3 and water, dried
(MgSO4) and evaporated in-vacuo. The residue was purified by radial
chromatography on silica eluting with 5% MeOH/DCM to yield the
product as a gum (17 mg). MS m/z ([M+H].sup.+) 677.2.
[0286] Intermediates S, T and U were prepared by the general method
described for Intermediate R using the appropriate amine and
substituting NaBH.sub.4 for NaBH.sub.3CN.
Intermediate S
[0287] Intermediate Q (40 mg) yielded Intermediate S (21 mg).
[0288] MS m/z ([M+H].sup.+) 693.1.
Intermediate T
[0289] Intermediate Q (62 mg) yielded Intermediate T (42 mg).
[0290] MS m/z ([M+H].sup.+) 691.2, ([2M+H].sup.+) 1381.0
Intermediate U
[0291] Intermediate Q (62 mg) yielded Intermediate U (28 mg).
[0292] MS m/z ([M+H].sup.+) 692.1.
Example 11
[0293] Intermediate R (10 mg) was stirred with 7M NH.sub.3 in MeOH
(1 mL) at ambient temperature for 18 hrs. The reaction was
evaporated to dryness in-vacuo and the residue dissolved in water
and washed with DCM. The aqueous layer was then subjected directly
to preparative HPLC. This process afforded the target compound as
an oil (1.3 mg).
[0294] MS m/z ([M+H].sup.+) 365.1, ([2M+H].sup.+) 728.8.
[0295] 1H .sup.1H NMR (CD.sub.3OD) .delta. 7.69 (d, 1H), 6.34 (d,
1H), 5.92 (s, 1H), 5.20 (d, 1H), 4.31 (d, 1H), 3.98-3.90 (m, 1H),
3.80-3.65 (m, 3H), 3.50-3.35 (m, 4H), 1.98-1.90 (m, 4H), 1.20 (s,
3H).
[0296] The general method for Example 11 was applied to the
preparation of Examples 12-14.
Example 12
[0297] Intermediate S (20 mg) yielded compound 12 (2.4 mg) after
purification by preparative HPLC.
[0298] MS m/z ([M+H].sup.+) 381.1, ([2M+Na].sup.+) 782.8.
[0299] 1H .sup.1H NMR (CD.sub.3OD) .delta. 7.97 (d, 1H), 6.68 (d,
1H), 5.96 (s, 1H), 4.48 (d, 1H), 4.12 (d, 1H), 3.96-3.71 (m, 8H),
3.27-2.87 (m, 4H), 1.12 (s, 3H).
Example 13
[0300] Intermediate T (42 mg) yielded 4.27 mg compound 13 (4.3 mg)
after purification by preparative HPLC.
[0301] MS m/z ([M+H].sup.+) 379.2, ([2M+H].sup.+) 756.9.
[0302] 1H .sup.1H NMR (CD.sub.3OD) .delta. 7.93 (d, 1H), 6.65 (d,
1H), 5.97 (s, 1H), 4.42 (d, 1H), 4.07 (d, 1H), 3.93 (d, 1H),
3.82-3.71 (m, 3H), 3.04-2.87 (m, 4H), 1.86-1.60 (m, 6H), 1.13 (s,
3H).
Example 14
[0303] Intermediate U (27 mg) yielded compound 14 (2.1 mg) after
purification by preparative HPLC.
[0304] MS m/z ([M+H].sup.+) 380.1, ([2M+Na].sup.+) 780.8.
[0305] 1H .sup.1H NMR (CD.sub.3OD) .delta. 8.08 (d, 1H), 6.69 (d,
1H), 5.96, (s, 1H), 4.51 (d, 1H), 4.10 (d, 1H), 3.94-3.51 (m, 6H),
3.27-2.87 (m, 6H), 1.14 (s, 3H).
Example 15
[0306] Example 1 (31 mg) was dried overnight under high vacuum over
P.sub.2O.sub.5 and then stirred with trimethyl phosphate (1.7 mL)
at ambient temperature with oven-dried molecular sieves (4 .ANG.)
for 16 hrs under argon. The reaction was cooled to 0.degree. C.,
POCl.sub.3 (31 .mu.l) added and stirring continued for 2 hours
before addition of Bu.sub.3N (72 .mu.l) followed by acetonitrile
(0.5 mL) and tributylammonium pyrophosphate (190 mg). After a
further 2 hrs at 0.degree. C. the reaction was quenched by pouring
into ice-cold 1M triethylammonium bicarbonate buffer (10 mL, pH
8.5) and the aqueous layer washed with Et.sub.2O (3.times.10 mL).
The aqueous material was then lyophilised to give a white solid
which was purified by preparative HPLC to afford the product (1.67
mg).
[0307] MS m/z ([M-H].sup.-) 547.6, ([M+3TEA+H].sup.+) 852.5.
[0308] .sup.31P NMR (D.sub.2O) .delta. -8.14 (m, 1P), -10.69 (m,
1P), -21.65 (m, 1P).
Biological Data
In Vitro Anti-HCV Dose-Response and Cytotoxicity
HCV Replicon Assay
[0309] The antiviral activity of test compounds were assayed in the
stable HCV RNA-replicating cell line, AVA5, derived by transfection
of the human hepatoblastoma cell line, Huh7 (Blight, et al., 2000,
Sci. 290:1972). Concentrations of compounds were added to dividing
cultures once daily for three days and intracellular HCV RNA levels
and cytotoxicity assessed 24 hours after the last dose of
compound.
[0310] Intracellular HCV RNA levels were measured using standard
blot hybridization techniques using triplicate cultures and levels
of .beta.-actin RNA were used to normalize HCV RNA levels in each
sample. Cytotoxicity was measured using an established neutral red
dye uptake assay (B. E. Korba and J. L. Gerin. 1992. Antivir. Res.
19, 55-70) and the 50% effective antiviral concentrations
(EC.sub.50) and cytotoxic concentrations (CC.sub.50) were
calculated using a computer program for curve fitting.
[0311] Examples such as 1-14 were typically active in the replicon
assay in the range 1 to >1000 .mu.M and cytotoxic in the range
30 to >100 .mu.M.
HCV Polymerase Inhibition Assay
[0312] HCV 3'UTR RNA template was synthesized, gel purified and
quantified by spectrophotometry. The kinetic constant, K.sub.m, was
determined for the RNA template and for each GTP, CTP, ATP and UTP
using a non-linear least square fit of initial rates as a function
of substrate concentration assuming Michaelis-Menten kinetics.
[0313] Standard RdRp assays consisted of 30 nM RNA template and 25
nM HCV NS5b.DELTA.21 (genotype 1b) (Replizyme Ltd) in a 50 L
reaction mixture containing 20 mM Tris-HCl, 5 mM MgCl.sub.2, 3 mM
DTT, 0.05% BSA, 22 nM GTP, 600 nM ATP, 15 nM CTP, 30 nM UTP and 3
nM [.alpha.-.sup.33P]GTP. Elongation reactions were initiated by
the addition of NTPs and proceeded for 30 mins at 25.degree. C.
Reactions were quenched by the addition of 0.2 M EDTA and product
formation was collected by filtration through Multiscreen plates
(Millipore). Quantification of product formation was performed
using TopCount (Perkin Elmer).
[0314] The inhibitor concentration at which the enzyme catalyzed
rate is reduced by half (IC.sub.50) was determined using a computer
program for curve fitting
[0315] Examples such as 15 were typically inhibitory of NS5b in the
range 100 to >1000 .mu.M.
[0316] The reference to any prior art in this specification is not,
and should not be taken as, an acknowledgment or any form of
suggestion that that prior art forms part of the common general
knowledge.
[0317] Throughout this specification and the claims which follow,
unless the context requires otherwise, the word "comprise", and
variations such as "comprises" and "comprising", will be understood
to imply the inclusion of a stated integer or step or group of
integers or steps but not the exclusion of any other integer or
step or group of integers or steps.
* * * * *