U.S. patent application number 12/271388 was filed with the patent office on 2009-12-24 for 2',4'-substituted nucleosides as antiviral agents.
This patent application is currently assigned to PHARMASSET, INC.. Invention is credited to JINFA DU, MICHAEL JOSEPH SOFIA.
Application Number | 20090318380 12/271388 |
Document ID | / |
Family ID | 40373557 |
Filed Date | 2009-12-24 |
United States Patent
Application |
20090318380 |
Kind Code |
A1 |
SOFIA; MICHAEL JOSEPH ; et
al. |
December 24, 2009 |
2',4'-SUBSTITUTED NUCLEOSIDES AS ANTIVIRAL AGENTS
Abstract
Embodiments of the invention are to compounds, methods, and
compositions for use in the treatment of viral infections. More
specifically embodiments of the invention are 2',4'-substituted
nucleoside compounds useful for the treatment of viral infections,
such as HIV, HCV, and HBV infections.
Inventors: |
SOFIA; MICHAEL JOSEPH;
(DOYLESTOWN, PA) ; DU; JINFA; (NEW HOPE,
PA) |
Correspondence
Address: |
MERCHANT & GOULD PC
P.O. BOX 2903
MINNEAPOLIS
MN
55402-0903
US
|
Assignee: |
PHARMASSET, INC.
PRINCETON
NJ
|
Family ID: |
40373557 |
Appl. No.: |
12/271388 |
Filed: |
November 14, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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60989296 |
Nov 20, 2007 |
|
|
|
Current U.S.
Class: |
514/45 ; 514/49;
514/50; 534/551; 536/27.2; 536/27.21; 536/27.6; 536/28.2;
536/28.5 |
Current CPC
Class: |
A61P 31/20 20180101;
C07H 19/173 20130101; A61K 31/7076 20130101; C07H 19/06 20130101;
C07H 19/073 20130101; A61K 31/7064 20130101; A61P 31/04 20180101;
C07H 19/14 20130101; A61K 31/7072 20130101; A61P 31/12 20180101;
A61P 31/18 20180101; A61P 31/00 20180101; C07H 19/16 20130101; A61P
31/14 20180101; A61P 35/00 20180101; A61K 31/7068 20130101 |
Class at
Publication: |
514/45 ;
536/27.6; 536/27.21; 536/27.2; 536/28.2; 536/28.5; 514/50; 514/49;
534/551 |
International
Class: |
A61K 31/7068 20060101
A61K031/7068; C07H 19/16 20060101 C07H019/16; C07H 19/06 20060101
C07H019/06; A61K 31/7076 20060101 A61K031/7076; A61K 31/7064
20060101 A61K031/7064 |
Claims
1. A compound of the formula: ##STR00124## wherein for structure A
or A' (a) R.sup.2 is independently CH.sub.3, CH.sub.2F, CHF.sub.2,
CF.sub.3, F, CN, C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, or C.sub.1-4
alkyl optionally substituted with amino, hydroxy, or 1 to 3
fluorine atoms; (b) R is H, phosphate, including 5'-monophosphate,
5',3'-cyclic phosphate, diphosphate, triphosphate, or a stabilized
phosphate prodrug, H-phosphonate, including stabilized
H-phosphonates, acyl, including optionally substituted phenyl and
lower acyl, alkyl, including lower alkyl, O-substituted
carboxyalkylamino or its peptide derivatives, sulfonate ester,
including alkyl or arylalkyl sulfonyl, including methanesulfonyl
and benzyl, wherein the phenyl group is optionally substituted, a
lipid, including a phospholipid, an L or D-amino acid, a
carbohydrate, a peptide, a cholesterol, or other pharmaceutically
acceptable leaving group which when administered in vivo; (c)
R.sup.3 is independently OH, H, C.sub.1-4 alkyl, C.sub.2-4 alkenyl,
C.sub.2-4 alkynyl, vinyl, N.sub.3, CN, Cl, Br, F, I, NO.sub.2,
C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.2-4
alkynyl), C(O)O(C.sub.2-4 alkenyl), O(C.sub.1-10 acyl), O(C.sub.1-4
alkyl), O(C.sub.2-4 alkenyl), SH, S(C.sub.1-4 acyl), S(C.sub.1-4
alkyl), S(C.sub.2-4 alkynyl), S(C.sub.2-4 alkenyl), SO(C.sub.1-4
acyl), SO(C.sub.1-4 alkyl), SO(C.sub.2-4 alkynyl), SO(C.sub.2-4
alkenyl), SO.sub.2(C.sub.1-4 acyl), SO.sub.2(C.sub.1-4 alkyl),
SO.sub.2(C.sub.2-4 alkynyl), SO.sub.2(C.sub.2-4 alkenyl),
OS(O).sub.2(C.sub.1-4 acyl), OS(O).sub.2(C.sub.1-4 alkyl),
OS(O).sub.2(C.sub.2-4 alkenyl), NH.sub.2, NH(C.sub.1-4 alkyl),
NH(C.sub.2-4 alkenyl), NH(C.sub.2-4 alkynyl), NH(C.sub.1-4 acyl),
N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-18 acyl).sub.2, wherein alkyl,
alkynyl, alkenyl and vinyl are optionally substituted by N.sub.3,
CN, one to three halogen (Cl, Br, F, I), NO.sub.2, C(O)O(C.sub.1-4
alkyl), C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.2-4 alkynyl),
C(O)O(C.sub.2-4 alkenyl), O(C.sub.1-4 acyl), O(C.sub.1-4 alkyl),
O(C.sub.2-4 alkenyl), SH, S(C.sub.1-4 acyl), S(C.sub.1-4 alkyl),
S(C.sub.2-4 alkynyl), S(C.sub.2-4 alkenyl), SO(C.sub.1-4 acyl),
SO(C.sub.1-4 alkyl), SO(C.sub.2-4 alkynyl), SO(C.sub.2-4 alkenyl),
SO.sub.2(C.sub.1-4 acyl), SO.sub.2(C.sub.1-4 alkyl),
SO.sub.2(C.sub.2-4 alkynyl), SO.sub.2(C.sub.2-4 alkenyl),
OS(O).sub.2(C.sub.1-4 acyl), OS(O).sub.2(C.sub.14 alkyl),
OS(O).sub.2(C.sub.2-4 alkenyl), NH.sub.2, NH(C.sub.1-4 alkyl),
NH(C.sub.2-4 alkenyl), NH(C.sub.2-4 alkynyl), NH(C.sub.1-4 acyl),
N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-4 acyl).sub.2; (d) R.sup.4 is
independently H, a lower alkyl, CN, vinyl, O-(lower alkyl),
hydroxyl lower alkyl, i.e., --(CH.sub.2).sub.pOH, where p is 1-6,
including hydroxylmethyl (CH.sub.2OH), CH.sub.2F, N.sub.3,
CH.sub.2CN, CH.sub.2NH.sub.2, CH.sub.2NHCH.sub.3,
CH.sub.2N(CH.sub.3).sub.2, ethynyl alkyne (optionally substituted),
or halogen, including F, Cl, Br, or I, alkenyl, alkynyl, Br-vinyl,
hydroxy, O-alkenyl, NO.sub.2, amino, loweralkylamino, or
di(loweralkyl)amino; (e) R and R.sup.3 can together form
5',3'-cyclic phosphate including stabilized prodrugs thereof; (f)
Base is a naturally occurring or modified purine or pyrimidine base
represented by the following structures: ##STR00125## wherein for a
and b Z is Nor CR.sup.8; R.sup.5, R.sup.6, and R.sup.7 are
independently H, F, Cl, Br, I, OH, OR', SH, SR', NH.sub.2, NHR',
NR'.sub.2 (two R' can form saturated or unsaturated rings, or
saturated or unsaturated heterocyclic rings), lower alkyl of
C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkyl of
C.sub.1-C.sub.6, lower alkenyl of C.sub.2-C.sub.6, halogenated (F,
Cl, Br, I) lower alkenyl of C.sub.2-C.sub.6, lower alkynyl of
C.sub.2-C.sub.6 such as C--CH, halogenated (F, Cl, Br, I) lower
alkynyl of C.sub.2-C.sub.6, lower alkoxy of C.sub.1-C.sub.6,
halogenated (F, Cl, Br, I) lower alkoxy of C.sub.1-C.sub.6,
CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR', CONR.sub.12,
CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R' wherein R' is an
optionally substituted alkyl, which includes, but is not limited
to, an optionally substituted C.sub.1-20 alkyl, an optionally
substituted C.sub.1-10 alkyl, an optionally substituted lower
alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkylaryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; R.sup.8 is
independently H, halogen (including F, Cl, Br, I), OH, OR', SH,
SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form saturated or
unsaturated rings, or saturated or unsaturated heterocyclic rings),
NO.sub.2, lower alkyl of C.sub.1-C.sub.6, halogenated (F, Cl, Br,
I) lower alkyl of C.sub.1-C.sub.6, lower alkenyl of
C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower alkenyl of
C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6, halogenated (F,
Cl, Br, I) lower alkynyl of C.sub.2-C.sub.6, lower alkoxy of
C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkoxy of
C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR',
CONR.sub.12, CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R', wherein R'
is an optionally substituted alkyl, which includes, but is not
limited to, an optionally substituted C.sub.1-20 alkyl, an
optionally substituted C.sub.1-10 alkyl, an optionally substituted
lower alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkyl-aryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; or base may be
selected from a group of formula c ##STR00126## wherein for
structure c, 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, CNNR, 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.sup.X 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 linesindicate a possible pi
or double bond; 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; or base may be
a structure selected from the group consisting of structures d-n
##STR00127## ##STR00128## wherein Z, X, and R'' are defined as in
structure c; base may be a structure selected from the group
consisting of structures o-ff ##STR00129## ##STR00130##
##STR00131## ##STR00132## wherein G and R are defined as in
structure c; base may be a structure gg ##STR00133## 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 defined as in
structure c; base may be a structure hh ##STR00134## 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 in 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 defined as in structure c;
base may be a structure ii ##STR00135## wherein R and G are defined
as in structure c; base may be a structure jj ##STR00136## wherein
R and G are defined as in structure c; or base may be a structure
kk ##STR00137## wherein for structure kk: R is selected from the
group consisting of hydrogen and C.sub.1-C.sub.3 alkyl; X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
Y is selected from the group consisting of a bond, O, and CH.sub.2;
Q is absent or is selected from the group consisting of O, S, and
NH, provided that when Q is absent, V and NH are both attached to a
CH.sub.2 group; V is selected from the group consisting of N and
C-G; Z is selected from the group consisting of N and C-G'; G and
G' are independently selected from the group consisting of
hydrogen, amino, aminocarbonyl, methylamino, dimethylamino,
acylamino, alkoxyamino, --SO.sub.3H, --SO.sub.2NH.sub.2,
aminocarbonylamino, oxycarbonylamino, HR'NCHR''C(O)NH--, azido,
cyano, halo, hydroxyamino, and hydrazino, where R' is hydrogen and
R'' is a side-chain of an amino acid or where R' and R'' together
with the nitrogen and carbon bound to each group respectively form
a pyrrolidinyl group; provided that V and Z are not identical;
provided that when V is C--H, Z is N; T.sup.1 and T.sup.2 are
independently selected from the group consisting of hydrogen,
hydroxyl, C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy,
amino, substituted amino, and halo; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may
be a structure ll ##STR00138## wherein for structure ll: R is
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; Q' is selected from the group
consisting of NH, O, and S; G' is selected from the group
consisting of amino, aminocarbonyl, methylamino, dimethylamino,
acylamino, --SO.sub.3H, --SO.sub.2NH.sub.2, alkoxyamino,
aminocarbonylamino, oxycarbonylamino, HR'NCHR''C(O)NH--, azido,
cyano, halo, hydroxyamino, and hydrazino, where R' is hydrogen and
R'' is a side-chain of an amino acid or where R' and R'' together
with the nitrogen and carbon bound to each group respectively form
a pyrrolidinyl group; Y is selected from the group consisting of a
bond, O, and CH.sub.2; and each of W, W.sup.1, and W.sup.2 is
independently selected from the group consisting of hydrogen,
C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may be a
structure mm ##STR00139## where in for structure mm Q is as defined
for structure kk A and B are independently selected from the group
consisting of C=Q, NH, and methylene optionally substituted with 1
to 2 halo groups, provided that A and B are not both NH; D is NH,
or -D-A-B- together form a --N.dbd.CH--NH--,
--(C=Q)-CH.sub.2--(C=Q)-, --(C=Q)-NH--(C=Q)-,
--(CX')=(CX')--(C=Q)-, or --CH.dbd.CH--NH-- group where X' is halo;
each Q is independently selected from the group consisting of O, S,
and NH; R is selected from the group consisting of hydrogen and
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; T.sup.1 and T.sup.2 are independently
selected from the group consisting of hydrogen, hydroxyl,
C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy, amino,
substituted amino, and halo; Y is selected from the group
consisting of a bond, O, and CH.sub.2; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; and
pharmaceutically acceptable salts, tautomers, pharmaceutically
acceptable salts of tautomers, salts (acidic or basic addition
salts), hydrates, solvates, crystalline forms thereof, optionally
in combination with one or more antiviral, antibacterial, or
antiproliferative agents.
2. A compound of the formula of claim 1 or its pharmaceutically
acceptable salt.
3. A pharmaceutical composition for treating HBV, HCV or HIV that
includes a pharmaceutically effective treatment amount of a
compound of claim 1 in a pharmaceutically acceptable carrier or
diluent.
4. A compound according to claim 1, wherein the compound is
represented by formula A.
5. A compound according to claim 1 wherein the compound is
##STR00140##
6. A method for the treatment of a host infected with HBV, HCV, or
HIV that includes administering an effective amount of a compound
of the formula: ##STR00141## wherein for structure A or A' (a)
R.sup.2 is independently CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3,
F, CN, C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, or C.sub.1-4 alkyl
optionally substituted with amino, hydroxy, or 1 to 3 fluorine
atoms; (b) R is H, phosphate, including 5'-monophosphate,
5',3'-cyclic phosphate, diphosphate, triphosphate, or a stabilized
phosphate prodrug, H-phosphonate, including stabilized
H-phosphonates, acyl, including optionally substituted phenyl and
lower acyl, alkyl, including lower alkyl, O-substituted
carboxyalkylamino or its peptide derivatives, sulfonate ester,
including alkyl or arylalkyl sulfonyl, including methanesulfonyl
and benzyl, wherein the phenyl group is optionally substituted, a
lipid, including a phospholipid, an L or D-amino acid, a
carbohydrate, a peptide, a cholesterol, or other pharmaceutically
acceptable leaving group which when administered in vivo; (c)
R.sup.3 is independently OH, H, C.sub.1-4 alkyl, C.sub.2-4 alkenyl,
C.sub.2-4 alkynyl, vinyl, N.sub.3, CN, Cl, Br, F, I, NO.sub.2,
C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.2-4
alkynyl), C(O)O(C.sub.2-4 alkenyl), O(C.sub.1-10 acyl), O(C.sub.1-4
alkyl), O(C.sub.2-4 alkenyl), SH, S(C.sub.1-4 acyl), S(C.sub.1-4
alkyl), S(C.sub.2-4 alkynyl), S(C.sub.2-4 alkenyl), SO(C.sub.1-4
acyl), SO(C.sub.1-4 alkyl), SO(C.sub.2-4 alkynyl), SO(C.sub.2-4
alkenyl), SO.sub.2(C.sub.1-4 acyl), SO.sub.2(C.sub.1-4 alkyl),
SO.sub.2(C.sub.2-4 alkynyl), SO.sub.2(C.sub.2-4 alkenyl),
OS(O).sub.2(C.sub.1-4 acyl), OS(O).sub.2(C.sub.1-4 alkyl),
OS(O).sub.2(C.sub.2-4 alkenyl), NH.sub.2, NH(C.sub.1-4 alkyl),
NH(C.sub.2-4 alkenyl), NH(C.sub.2-4 alkynyl), NH(C.sub.1-4 acyl),
N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-18 acyl).sub.2, wherein alkyl,
alkynyl, alkenyl and vinyl are optionally substituted by N.sub.3,
CN, one to three halogen (Cl, Br, F, I), NO.sub.2, C(O)O(C.sub.1-4
alkyl), C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.2-4 alkynyl),
C(O)O(C.sub.2-4 alkenyl), O(C.sub.1-4 acyl), O(C.sub.1-4 alkyl),
O(C.sub.2-4 alkenyl), SH, S(C.sub.1-4 acyl), S(C.sub.1-4 alkyl),
S(C.sub.2-4 alkynyl), S(C.sub.2-4 alkenyl), SO(C.sub.1-4 acyl),
SO(C.sub.1-4 alkyl), SO(C.sub.2-4 alkynyl), SO(C.sub.2-4 alkenyl),
SO.sub.2(C.sub.1-4 acyl), SO.sub.2(C.sub.1-4 alkyl),
SO.sub.2(C.sub.2-4 alkynyl), SO.sub.2(C.sub.2-4 alkenyl),
OS(O).sub.2(C.sub.1-4 acyl), OS(O).sub.2(C.sub.1-4 alkyl),
OS(O).sub.2(C.sub.2-4 alkenyl), NH.sub.2, NH(C.sub.1-4 alkyl),
NH(C.sub.2-4 alkenyl), NH(C.sub.2-4 alkynyl), NH(C.sub.1-4 acyl),
N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-4 acyl).sub.2; (d) R.sup.4 is
independently H, a lower alkyl, CN, vinyl, O-(lower alkyl),
hydroxyl lower alkyl, i.e., --(CH.sub.2).sub.pOH, where p is 1-6,
including hydroxylmethyl (CH.sub.2OH), CH.sub.2F, N.sub.3,
CH.sub.2CN, CH.sub.2NH.sub.2, CH.sub.2NHCH.sub.3,
CH.sub.2N(CH.sub.3).sub.2, ethynyl alkyne (optionally substituted),
or halogen, including F, Cl, Br, or I, alkenyl, alkynyl, Br-vinyl,
hydroxy, O-alkenyl, NO.sub.2, amino, loweralkylamino, or
di(loweralkyl)amino; (e) R and R.sup.3 can together form
5',3'-cyclic phosphate including stabilized prodrugs thereof, (f)
Base is a naturally occurring or modified purine or pyrimidine base
represented by the following structures: ##STR00142## wherein for a
and b Z is Nor CR.sup.8; R.sup.5, R.sup.6, and R.sup.7 are
independently H, F, Cl, Br, I, OH, OR', SH, SR', NH.sub.2, NHR',
NR'.sub.2 (two R' can form saturated or unsaturated rings, or
saturated or unsaturated heterocyclic rings), lower alkyl of
C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkyl of
C.sub.1-C.sub.6, lower alkenyl of C.sub.2-C.sub.6, halogenated (F,
Cl, Br, I) lower alkenyl of C.sub.2-C.sub.6, lower alkynyl of
C.sub.2-C.sub.6 such as C--CH, halogenated (F, Cl, Br, I) lower
alkynyl of C.sub.2-C.sub.6, lower alkoxy of C.sub.1-C.sub.6,
halogenated (F, Cl, Br, I) lower alkoxy of C.sub.1-C.sub.6,
CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR', CONR.sub.12,
CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R', wherein R' is an
optionally substituted alkyl, which includes, but is not limited
to, an optionally substituted C.sub.1-20 alkyl, an optionally
substituted C.sub.1-10 alkyl, an optionally substituted lower
alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkyl-aryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; R.sup.8 is
independently H, halogen (including F, Cl, Br, I), OH, OR', SH,
SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form saturated or
unsaturated rings, or saturated or unsaturated heterocyclic rings),
NO.sub.2, lower alkyl of C.sub.1-C.sub.6, halogenated (F, Cl, Br,
I) lower alkyl of C.sub.1-C.sub.6, lower alkenyl of
C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower alkenyl of
C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6, halogenated (F,
Cl, Br, I) lower alkynyl of C.sub.2-C.sub.6, lower alkoxy of
C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkoxy of
C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR',
CONR.sub.12, CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R', wherein R'
is an optionally substituted alkyl, which includes, but is not
limited to, an optionally substituted C.sub.1-20 alkyl, an
optionally substituted C.sub.1-10 alkyl, an optionally substituted
lower alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkyl-aryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; or base may be
selected from a group of formula c ##STR00143## wherein for
structure c, 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, CNNR, 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.sup.X 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 linesindicate a possible pi or
double bond; 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; or base may be
a structure selected from the group consisting of structures d-n
##STR00144## ##STR00145## wherein Z, X, and R'' are defined as in
structure c; base may be a structure selected from the group
consisting of structures o-ff ##STR00146## ##STR00147##
##STR00148## ##STR00149## wherein G and R are defined as in
structure c; base may be a structure gg ##STR00150## 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 defined as in
structure c; base may be a structure hh ##STR00151## 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 in 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 defined as in structure c;
base may be a structure ii ##STR00152## wherein R and G are defined
as in structure c; base may be a structure jj ##STR00153## wherein
R and G are defined as in structure c; or base may be a structure
kk ##STR00154## wherein for structure kk: R is selected from the
group consisting of hydrogen and C.sub.1-C.sub.3 alkyl; X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
Y is selected from the group consisting of a bond, O, and CH.sub.2;
Q is absent or is selected from the group consisting of O, S, and
NH, provided that when Q is absent, V and NH are both attached to a
CH.sub.2 group; V is selected from the group consisting of N and
C-G; Z is selected from the group consisting of N and C-G'; G and
G' are independently selected from the group consisting of
hydrogen, amino, aminocarbonyl, methylamino, dimethylamino,
acylamino, alkoxyamino, --SO.sub.3H, --SO.sub.2NH.sub.2,
aminocarbonylamino, oxycarbonylamino, HR'NCHR''C(O)NH--, azido,
cyano, halo, hydroxyamino, and hydrazino, where R' is hydrogen and
R'' is a side-chain of an amino acid or where R' and R'' together
with the nitrogen and carbon bound to each group respectively form
a pyrrolidinyl group; provided that V and Z are not identical;
provided that when V is C--H, Z is N; T.sup.1 and T.sup.2 are
independently selected from the group consisting of hydrogen,
hydroxyl, C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy,
amino, substituted amino, and halo; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may
be a structure ll ##STR00155## wherein for structure ll: R is
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; Q' is selected from the group
consisting of NH, O, and S; G' is selected from the group
consisting of amino, aminocarbonyl, methylamino, dimethylamino,
acylamino, --SO.sub.3H, --SO.sub.2NH.sub.2, alkoxyamino,
aminocarbonylamino, oxycarbonylamino, HR'NCHR''C(O)NH--, azido,
cyano, halo, hydroxyamino, and hydrazino, where R' is hydrogen and
R'' is a side-chain of an amino acid or where R' and R'' together
with the nitrogen and carbon bound to each group respectively form
a pyrrolidinyl group; Y is selected from the group consisting of a
bond, O, and CH.sub.2; and each of W, W.sup.1, and W.sup.2 is
independently selected from the group consisting of hydrogen,
C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may be a
structure mm ##STR00156## where in for structure mm Q is as defined
for structure kk A and B are independently selected from the group
consisting of C=Q, NH, and methylene optionally substituted with 1
to 2 halo groups, provided that A and B are not both NH; D is NH,
or -D-A-B- together form a --N.dbd.CH--NH--,
--(C=Q)-CH.sub.2--(C=Q)-, --(C=Q)-NH--(C=Q)-,
--(CX')=(CX')--(C=Q)-, or --CH.dbd.CH--NH-- group where X' is halo;
each Q is independently selected from the group consisting of O, S,
and NH; R is selected from the group consisting of hydrogen and
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; T.sup.1 and T.sup.2 are independently
selected from the group consisting of hydrogen, hydroxyl,
C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy, amino,
substituted amino, and halo; Y is selected from the group
consisting of a bond, O, and CH.sub.2; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; and
pharmaceutically acceptable salts, tautomers, pharmaceutically
acceptable salts of tautomers, salts (acidic or basic addition
salts), hydrates, solvates, crystalline forms thereof, optionally
in combination with one or more antiviral, antibacterial, or
antiproliferative agents.
7. A use of the compound represented by formula A or A' in the
manufacture of a medicament for the treatment and/or prophylaxis of
any condition the result of an infection by HBV, HCV, or HIV:
##STR00157## wherein for structure A or A' (a) R.sup.2 is
independently CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, F, CN,
C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, or C.sub.1-4 alkyl optionally
substituted with amino, hydroxy, or 1 to 3 fluorine atoms; (b) R is
H, phosphate, including 5'-monophosphate, 5',3'-cyclic phosphate,
diphosphate, triphosphate, or a stabilized phosphate prodrug,
H-phosphonate, including stabilized H-phosphonates, acyl, including
optionally substituted phenyl and lower acyl, alkyl, including
lower alkyl, O-substituted carboxyalkylamino or its peptide
derivatives, sulfonate ester, including alkyl or arylalkyl
sulfonyl, including methanesulfonyl and benzyl, wherein the phenyl
group is optionally substituted, a lipid, including a phospholipid,
an L or D-amino acid, a carbohydrate, a peptide, a cholesterol, or
other pharmaceutically acceptable leaving group which when
administered in vivo; (c) R.sup.3 is independently OH, H, C.sub.1-4
alkyl, C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, vinyl, N.sub.3, CN,
Cl, Br, F, I, NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4
alkyl), C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl),
O(C.sub.1-10 acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH,
S(C.sub.1-4 acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl),
S(C.sub.2-4 alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl),
SO(C.sub.2-4 alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4
acyl), SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.1-4 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2,
N(C.sub.1-18 acyl).sub.2, wherein alkyl, alkynyl, alkenyl and vinyl
are optionally substituted by N.sub.3, CN, one to three halogen
(Cl, Br, F, I), NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4
alkyl), C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl),
O(C.sub.1-4 acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH,
S(C.sub.1-4 acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl),
S(C.sub.2-4 alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl),
SO(C.sub.2-4 alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4
acyl), SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.1-4 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-4
acyl).sub.2; (d) R.sup.4 is independently H, a lower alkyl, CN,
vinyl, O-(lower alkyl), hydroxyl lower alkyl, i.e.,
--(CH.sub.2).sub.pOH, where p is 1-6, including hydroxylmethyl
(CH.sub.2OH), CH.sub.2F, N.sub.3, CH.sub.2CN, CH.sub.2NH.sub.2,
CH.sub.2NHCH.sub.3, CH.sub.2N(CH.sub.3).sub.2, ethynyl alkyne
(optionally substituted), or halogen, including F, Cl, Br, or I,
alkenyl, alkynyl, Br-vinyl, hydroxy, O-alkenyl, NO.sub.2, amino,
loweralkylamino, or di(loweralkyl)amino; (e) R and R.sup.3 can
together form 5',3'-cyclic phosphate including stabilized prodrugs
thereof; (f) Base is a naturally occurring or modified purine or
pyrimidine base represented by the following structures:
##STR00158## wherein for a and b Z is N or CR.sup.8; R.sup.5,
R.sup.6, and R.sup.7 are independently H, F, Cl, Br, I, OH, OR',
SH, SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form saturated or
unsaturated rings, or saturated or unsaturated heterocyclic rings),
lower alkyl of C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower
alkyl of C.sub.1-C.sub.6, lower alkenyl of C.sub.2-C.sub.6,
halogenated (F, Cl, Br, I) lower alkenyl of C.sub.2-C.sub.6, lower
alkynyl of C.sub.2-C.sub.6 such as C--CH, halogenated (F, Cl, Br,
I) lower alkynyl of C.sub.2-C.sub.6, lower alkoxy of
C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkoxy of
C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR',
CONR.sub.12, CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R', wherein R'
is an optionally substituted alkyl, which includes, but is not
limited to, an optionally substituted C.sub.1-20 alkyl, an
optionally substituted C.sub.1-10 alkyl, an optionally substituted
lower alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkyl-aryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; R.sup.8 is
independently H, halogen (including F, Cl, Br, I), OH, OR', SH,
SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form saturated or
unsaturated rings, or saturated or unsaturated heterocyclic rings),
NO.sub.2, lower alkyl of C.sub.1-C.sub.6, halogenated (F, Cl, Br,
I) lower alkyl of C.sub.1-C.sub.6, lower alkenyl of
C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower alkenyl of
C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6, halogenated (F,
Cl, Br, I) lower alkynyl of C.sub.2-C.sub.6, lower alkoxy of
C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkoxy of
C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR',
CONR.sub.12, CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R', wherein R'
is an optionally substituted alkyl, which includes, but is not
limited to, an optionally substituted C.sub.1-20 alkyl, an
optionally substituted C.sub.1-10 alkyl, an optionally substituted
lower alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkyl-aryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; or base may be
selected from a group of formula c ##STR00159## wherein for
structure c, 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, CNNR, 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.sup.X 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; 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; or
base may be a structure selected from the group consisting of
structures d-n ##STR00160## ##STR00161## wherein Z, X, and R'' are
defined as in structure c; base may be a structure selected from
the group consisting of structures o-ff ##STR00162## ##STR00163##
##STR00164## ##STR00165## wherein G and R are defined as in
structure c; base may be a structure gg ##STR00166## 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 defined as in
structure c; base may be a structure hh ##STR00167## 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 in 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 defined as in structure c;
base may be a structure ii ##STR00168## wherein R and G are defined
as in structure c; base may be a structure jj ##STR00169## wherein
R and G are defined as in structure c; or base may be a structure
kk ##STR00170## wherein for structure kk: R is selected from the
group consisting of hydrogen and C.sub.1-C.sub.3 alkyl; X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
Y is selected from the group consisting of a bond, O, and CH.sub.2;
Q is absent or is selected from the group consisting of O, S, and
NH, provided that when Q is absent, V and NH are both attached to a
CH.sub.2 group; V is selected from the group consisting of N and
C-G; Z is selected from the group consisting of N and C-G'; G and
G' are independently selected from the group consisting of
hydrogen, amino, aminocarbonyl, methylamino, dimethylamino,
acylamino, alkoxyamino, --SO.sub.3H, --SO.sub.2NH.sub.2,
aminocarbonylamino, oxycarbonylamino, HR'NCHR''C(O)NH--, azido,
cyano, halo, hydroxyamino, and hydrazino, where R' is hydrogen and
R'' is a side-chain of an amino acid or where R' and R'' together
with the nitrogen and carbon bound to each group respectively form
a pyrrolidinyl group; provided that V and Z are not identical;
provided that when V is C--H, Z is N; T.sup.1 and T.sup.2 are
independently selected from the group consisting of hydrogen,
hydroxyl, C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy,
amino, substituted amino, and halo; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may
be a structure ll ##STR00171## wherein for structure ll: R is
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; Q' is selected from the group
consisting of NH, O, and S; G' is selected from the group
consisting of amino, aminocarbonyl, methylamino, dimethylamino,
acylamino, --SO.sub.3H, --SO.sub.2NH.sub.2, alkoxyamino,
aminocarbonylamino, oxycarbonylamino, HR'NCHR''C(O)NH--, azido,
cyano, halo, hydroxyamino, and hydrazino, where R' is hydrogen and
R'' is a side-chain of an amino acid or where R' and R'' together
with the nitrogen and carbon bound to each group respectively form
a pyrrolidinyl group; Y is selected from the group consisting of a
bond, O, and CH.sub.2; and each of W, W.sup.1, and W.sup.2 is
independently selected from the group consisting of hydrogen,
C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may be a
structure mm ##STR00172## where in for structure mm Q is as defined
for structure kk A and B are independently selected from the group
consisting of C=Q, NH, and methylene optionally substituted with 1
to 2 halo groups, provided that A and B are not both NH; D is NH,
or -D-A-B- together form a --N.dbd.CH--NH--,
--(C=Q)-CH.sub.2--(C=Q)-, --(C=Q)-NH--(C=Q)-,
--(CX')=(CX')--(C=Q)-, or --CH.dbd.CH--NH-- group where X' is halo;
each Q is independently selected from the group consisting of O, S,
and NH; R is selected from the group consisting of hydrogen and
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; T.sup.1 and T.sup.2 are independently
selected from the group consisting of hydrogen, hydroxyl,
C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy, amino,
substituted amino, and halo; Y is selected from the group
consisting of a bond, O, and CH.sub.2; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; and
pharmaceutically acceptable salts, tautomers, pharmaceutically
acceptable salts of tautomers, salts (acidic or basic addition
salts), hydrates, solvates, crystalline forms thereof; optionally
in combination with one or more antiviral, antibacterial, or
antiproliferative agents.
8. A method of making a medicament that comprises combining a
pharmaceutically effective carrier with a compound of the formula:
##STR00173## wherein for structure A or A' (a) R.sup.2 is
independently CH.sub.3, CH.sub.2F, CHF.sub.2, CF.sub.3, F, CN,
C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, or C.sub.1-4 alkyl optionally
substituted with amino, hydroxy, or 1 to 3 fluorine atoms; (b) R is
H, phosphate, including 5'-monophosphate, 5',3'-cyclic phosphate,
diphosphate, triphosphate, or a stabilized phosphate prodrug,
H-phosphonate, including stabilized H-phosphonates, acyl, including
optionally substituted phenyl and lower acyl, alkyl, including
lower alkyl, O-substituted carboxyalkylamino or its peptide
derivatives, sulfonate ester, including alkyl or arylalkyl
sulfonyl, including methanesulfonyl and benzyl, wherein the phenyl
group is optionally substituted, a lipid, including a phospholipid,
an L or D-amino acid, a carbohydrate, a peptide, a cholesterol, or
other pharmaceutically acceptable leaving group which when
administered in vivo; (c) R.sup.3 is independently OH, H, C.sub.1-4
alkyl, C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, vinyl, N.sub.3, CN,
Cl, Br, F, I, NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4
alkyl), C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl),
O(C.sub.1-10 acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH,
S(C.sub.1-4 acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl),
S(C.sub.2-4 alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl),
SO(C.sub.2-4 alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4
acyl), SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.1-4 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2,
N(C.sub.1-18 acyl).sub.2, wherein alkyl, alkynyl, alkenyl and vinyl
are optionally substituted by N.sub.3, CN, one to three halogen
(Cl, Br, F, I), NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4
alkyl), C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl),
O(C.sub.1-4 acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH,
S(C.sub.1-4 acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl),
S(C.sub.2-4 alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl),
SO(C.sub.2-4 alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4
acyl), SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.1-4 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-4
acyl).sub.2; (d) R.sup.4 is independently H, a lower alkyl, CN,
vinyl, O-(lower alkyl), hydroxyl lower alkyl, i.e.,
--(CH.sub.2).sub.pOH, where p is 1-6, including hydroxylmethyl
(CH.sub.2OH), CH.sub.2F, N.sub.3, CH.sub.2CN, CH.sub.2NH.sub.2,
CH.sub.2NHCH.sub.3, CH.sub.2N(CH.sub.3).sub.2, ethynyl alkyne
(optionally substituted), or halogen, including F, Cl, Br, or I,
alkenyl, alkynyl, Br-vinyl, hydroxy, O-alkenyl, NO.sub.2, amino,
loweralkylamino, or di(loweralkyl)amino; (e) R and R.sup.3 can
together form 5',3'-cyclic phosphate including stabilized prodrugs
thereof; (f) Base is a naturally occurring or modified purine or
pyrimidine base represented by the following structures:
##STR00174## wherein for a and b Z is N or CR.sup.8; R.sup.5,
R.sup.6, and R.sup.7 are independently H, F, Cl, Br, I, OH, OR',
SH, SR', NH.sub.2, NHR', NR'.sub.2, lower alkyl of C.sub.1-C.sub.6,
halogenated (F, Cl, Br, I) lower alkyl of C.sub.1-C.sub.6, lower
alkenyl of C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower
alkenyl of C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6 such
as C--CH, halogenated (F, Cl, Br, I) lower alkynyl of
C.sub.2-C.sub.6, lower alkoxy of C.sub.1-C.sub.6, halogenated (F,
Cl, Br, I) lower alkoxy of C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R',
CONH.sub.2, CONHR', CONR.sub.12, CH.dbd.CHCO.sub.2H, or
CH.dbd.CHCO.sub.2R' wherein R' is an optionally substituted alkyl,
which includes, but is not limited to, an optionally substituted
C.sub.1-20 alkyl, an optionally substituted C.sub.1-10 alkyl, an
optionally substituted lower alkyl; an optionally substituted
cycloalkyl; an optionally substituted alkynyl of C.sub.2-C.sub.6,
an optionally substituted lower alkenyl of C.sub.2-C.sub.6, or
optionally substituted acyl, which includes but is not limited to
C(O) alkyl, C(O)(C.sub.1-20 alkyl), C(O)(C.sub.1-10 alkyl), or
C(O)(lower alkyl), optionally substituted aryl, optionally
substituted C.sub.1-C.sub.4 alkyl-aryloxy, heteroaryl, optionally
substituted C.sub.1-C.sub.4 alkyl-heteroaryl, an optionally
substituted alkoxy C.sub.1-20 alkyl, an optionally substituted
amino C.sub.1-20 alkyl, an optionally substituted fluoro C.sub.1-20
alkyl; R.sup.8 is independently H, halogen (including F, Cl, Br,
I), OH, OR', SH, SR', NH.sub.2, NHR', NR'.sub.2, NO.sub.2, lower
alkyl of C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkyl of
C.sub.1-C.sub.6, lower alkenyl of C.sub.2-C.sub.6, halogenated (F,
Cl, Br, I) lower alkenyl of C.sub.2-C.sub.6, lower alkynyl of
C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower alkynyl of
C.sub.2-C.sub.6, lower alkoxy of C.sub.1-C.sub.6, halogenated (F,
Cl, Br, I) lower alkoxy of C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R',
CONH.sub.2, CONHR', CONR.sub.12, CH.dbd.CHCO.sub.2H, or
CH.dbd.CHCO.sub.2R', wherein R' is an optionally substituted alkyl,
which includes, but is not limited to, an optionally substituted
C.sub.1-20 alkyl, an optionally substituted C.sub.1-10 alkyl, an
optionally substituted lower alkyl; an optionally substituted
cycloalkyl; an optionally substituted alkynyl of C.sub.2-C.sub.6,
an optionally substituted lower alkenyl of C.sub.2-C.sub.6, or
optionally substituted acyl, which includes but is not limited to
C(O) alkyl, C(O)(C.sub.1-20 alkyl), C(O)(C.sub.1-10 alkyl), or
C(O)(lower alkyl), optionally substituted aryl, optionally
substituted C.sub.1-C.sub.4 alkyl-aryloxy, heteroaryl, optionally
substituted C.sub.1-C.sub.4 alkyl-heteroaryl, an optionally
substituted alkoxy C.sub.1-20 alkyl, an optionally substituted
amino C.sub.1-20 alkyl, an optionally substituted fluoro C.sub.1-20
alkyl; or base may be selected from a group of formula c
##STR00175## wherein for structure c, 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,
CNNR, 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.sup.X 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; 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; or base may be a structure selected from the
group consisting of structures d-n ##STR00176## ##STR00177##
wherein Z, X, and R'' are defined as in structure c; base may be a
structure selected from the group consisting of structures o-ff
##STR00178## ##STR00179## ##STR00180## ##STR00181## wherein G and R
are defined as in structure c; base may be a structure gg
##STR00182## 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 defined as in structure c; base may be a structure hh
##STR00183## 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
in 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 defined as in structure c; base may be a structure ii
##STR00184## wherein R and G are defined as in structure c; base
may be a structure jj ##STR00185## wherein R and G are defined as
in structure c; or base may be a structure kk ##STR00186## wherein
for structure kk: R is selected from the group consisting of
hydrogen and C.sub.1-C.sub.3 alkyl; X is selected from the group
consisting of hydrogen, halo, and OW.sup.2; Y is selected from the
group consisting of a bond, O, and CH.sub.2; Q is absent or is
selected from the group consisting of O, S, and NH, provided that
when Q is absent, V and NH are both attached to a CH.sub.2 group; V
is selected from the group consisting of N and C-G; Z is selected
from the group consisting of N and C-G'; G and G' are independently
selected from the group consisting of hydrogen, amino,
aminocarbonyl, methylamino, dimethylamino, acylamino, alkoxyamino,
--SO.sub.3H, --SO.sub.2NH.sub.2, aminocarbonylamino,
oxycarbonylamino, HR'NCHR''C(O)NH--, azido, cyano, halo,
hydroxyamino, and hydrazino, where R' is hydrogen and R'' is a
side-chain of an amino acid or where R' and R'' together with the
nitrogen and carbon bound to each group respectively form a
pyrrolidinyl group; provided that V and Z are not identical;
provided that when V is C--H, Z is N; T.sup.1 and T.sup.2 are
independently selected from the group consisting of hydrogen,
hydroxyl, C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy,
amino, substituted amino, and halo; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may
be a structure ll ##STR00187## wherein for structure ll: R is
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; Q' is selected from the group
consisting of NH, O, and S; G' is selected from the group
consisting of amino, aminocarbonyl, methylamino, dimethylamino,
acylamino, --SO.sub.3H, --SO.sub.2NH.sub.2, alkoxyamino,
aminocarbonylamino, oxycarbonylamino, HR'NCHR''C(O)NH--, azido,
cyano, halo, hydroxyamino, and hydrazino, where R' is hydrogen and
R'' is a side-chain of an amino acid or where R' and R'' together
with the nitrogen and carbon bound to each group respectively form
a pyrrolidinyl group; Y is selected from the group consisting of a
bond, O, and CH.sub.2; and each of W, W.sup.1, and W.sup.2 is
independently selected from the group consisting of hydrogen,
C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may be a
structure mm ##STR00188## where in for structure mm Q is as defined
for structure kk A and B are independently selected from the group
consisting of C=Q, NH, and methylene optionally substituted with 1
to 2 halo groups, provided that A and B are not both NH; D is NH,
or -D-A-B- together form a --N.dbd.CH--NH--,
--(C=Q)-CH.sub.2--(C=Q)-, --(C=Q)-NH--(C=Q)-,
--(CX')=(CX')--(C=Q)-, or --CH.dbd.CH--NH-- group where X' is halo;
each Q is independently selected from the group consisting of O, S,
and NH; R is selected from the group consisting of hydrogen and
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; T.sup.1 and T.sup.2 are independently
selected from the group consisting of hydrogen, hydroxyl,
C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy, amino,
substituted amino, and halo; Y is selected from the group
consisting of a bond, O, and CH.sub.2; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; and
pharmaceutically acceptable salts, tautomers, pharmaceutically
acceptable salts of tautomers, salts (acidic or basic addition
salts), hydrates, solvates, crystalline forms thereof; optionally
in combination with one or more antiviral, antibacterial, or
antiproliferative agents.
8. A compound selected from among the following compounds:
##STR00189## ##STR00190## ##STR00191## ##STR00192## ##STR00193##
##STR00194## ##STR00195## ##STR00196## ##STR00197## ##STR00198##
##STR00199## ##STR00200## ##STR00201## ##STR00202## ##STR00203##
##STR00204## ##STR00205## ##STR00206## ##STR00207## ##STR00208##
##STR00209##
9. A process, which comprises: (1) deoxygenating the 2'-C-position
of a compound represented by 1 or 1' ##STR00210## to obtain a
compound represented by 2 or 2' ##STR00211## and (2) derivatizing
the compound represented by 2 or 2' to obtain a compound
represented by A or A' ##STR00212## wherein for structures 1, 1',
2, 2', A, and A' (a) R.sup.2 is independently CH.sub.3, CH.sub.2F,
CHF.sub.2, CF.sub.3, F, CN, C.sub.2-4 alkenyl, C.sub.2-4 alkynyl,
or C.sub.1-4 alkyl optionally substituted with amino, hydroxy, or 1
to 3 fluorine atoms; (b) R is H, phosphate, including
5'-monophosphate, 5',3'-cyclic phosphate, diphosphate,
triphosphate, or a stabilized phosphate prodrug, H-phosphonate,
including stabilized H-phosphonates, acyl, including optionally
substituted phenyl and lower acyl, alkyl, including lower alkyl,
O-substituted carboxyalkylamino or its peptide derivatives,
sulfonate ester, including alkyl or arylalkyl sulfonyl, including
methanesulfonyl and benzyl, wherein the phenyl group is optionally
substituted, a lipid, including a phospholipid, an L or D-amino
acid, a carbohydrate, a peptide, a cholesterol, or other
pharmaceutically acceptable leaving group which when administered
in vivo; (c) R.sup.3 is independently OH, H, C.sub.1-4 alkyl,
C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, vinyl, N.sub.3, CN, Cl, Br,
F, I, NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4 alkyl),
C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl), O(C.sub.1-10
acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH, S(C.sub.1-4
acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl), S(C.sub.2-4
alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl), SO(C.sub.2-4
alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4 acyl),
SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.1-4 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2,
N(C.sub.1-18 acyl).sub.2, wherein alkyl, alkynyl, alkenyl and vinyl
are optionally substituted by N.sub.3, CN, one to three halogen
(Cl, Br, F, I), NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4
alkyl), C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl),
O(C.sub.1-4 acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH,
S(C.sub.1-4 acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl),
S(C.sub.2-4 alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl),
SO(C.sub.2-4 alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4
acyl), SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.14 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-4
acyl).sub.2; (d) R.sup.4 is independently H, a lower alkyl, CN,
vinyl, O-(lower alkyl), hydroxyl lower alkyl, i.e.,
--(CH.sub.2).sub.pOH, where p is 1-6, including hydroxylmethyl
(CH.sub.2OH), CH.sub.2F, N.sub.3, CH.sub.2CN, CH.sub.2NH.sub.2,
CH.sub.2NHCH.sub.3, CH.sub.2N(CH.sub.3).sub.2, ethynyl alkyne
(optionally substituted), or halogen, including F, Cl, Br, or I,
alkenyl, alkynyl, Br-vinyl, hydroxy, O-alkenyl, NO.sub.2, amino,
loweralkylamino, or di(loweralkyl)amino; (e) R and R3 can together
form 5',3'-cyclic phosphate including stabilized prodrugs thereof,
(f) Base is a naturally occurring or modified purine or pyrimidine
base represented by the following structures: ##STR00213## wherein
for a and b Z is Nor CR.sup.8; R.sup.5, R.sup.6, and R.sup.7 are
independently H, F, Cl, Br, I, OH, OR', SH, SR', NH.sub.2, NHR',
NR'.sub.2 (two R' can form saturated or unsaturated rings, or
saturated or unsaturated heterocyclic rings), lower alkyl of
C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkyl of
C.sub.1-C.sub.6, lower alkenyl of C.sub.2-C.sub.6, halogenated (F,
Cl, Br, I) lower alkenyl of C.sub.2-C.sub.6, lower alkynyl of
C.sub.2-C.sub.6 such as C.ident.CH, halogenated (F, Cl, Br, I)
lower alkynyl of C.sub.2-C.sub.6, lower alkoxy of C.sub.1-C.sub.6,
halogenated (F, Cl, Br, I) lower alkoxy of C.sub.1-C.sub.6,
CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR', CONR.sub.12,
CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R' wherein R' is an
optionally substituted alkyl, which includes, but is not limited
to, an optionally substituted C.sub.1-20 alkyl, an optionally
substituted C.sub.1-10 alkyl, an optionally substituted lower
alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkylaryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; R.sup.8 is
independently H, halogen (including F, Cl, Br, I), OH, OR', SH,
SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form saturated or
unsaturated rings, or saturated or unsaturated heterocyclic rings),
NO.sub.2, lower alkyl of C.sub.1-C.sub.6, halogenated (F, Cl, Br,
I) lower alkyl of C.sub.1-C.sub.6, lower alkenyl of
C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower alkenyl of
C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6, halogenated (F,
Cl, Br, I) lower alkynyl of C.sub.2-C.sub.6, lower alkoxy of
C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkoxy of
C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR',
CONR.sub.12, CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R', wherein R'
is an optionally substituted alkyl, which includes, but is not
limited to, an optionally substituted C.sub.1-20 alkyl, an
optionally substituted C.sub.1-10 alkyl, an optionally substituted
lower alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkyl-aryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; or base may be
selected from a group of formula c ##STR00214## wherein for
structure c, 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, CNNR, 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.sup.X 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 linesindicate a possible pi or
double bond; 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; or base may be
a structure selected from the group consisting of structures d-n
##STR00215## ##STR00216## wherein Z, X, and R'' are defined as in
structure c; base may be a structure selected from the group
consisting of structures o-ff ##STR00217## ##STR00218##
##STR00219## ##STR00220## wherein G and R are defined as in
structure c; base may be a structure gg ##STR00221## 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 defined as in
structure c; base may be a structure hh ##STR00222## 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 in 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 defined as in structure c;
base may be a structure ii ##STR00223## wherein R and G are defined
as in structure c; base may be a structure jj ##STR00224## wherein
R and G are defined as in structure c; or base may be a structure
kk ##STR00225## wherein for structure kk: R is selected from the
group consisting of hydrogen and C.sub.1-C.sub.3 alkyl; X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
Y is selected from the group consisting of a bond, O, and CH.sub.2;
Q is absent or is selected from the group consisting of O, S, and
NH, provided that when Q is absent, V and NH are both attached to a
CH.sub.2 group; V is selected from the group consisting of N and
C-G; Z is selected from the group consisting of N and C-G'; G and
G' are independently selected from the group consisting of
hydrogen, amino, aminocarbonyl, methylamino, dimethylamino,
acylamino, alkoxyamino, --SO.sub.3H, --SO.sub.2NH.sub.2,
aminocarbonylamino, oxycarbonylamino, HR'NCHR''C(O)NH--, azido,
cyano, halo, hydroxyamino, and hydrazino, where R' is hydrogen and
R'' is a side-chain of an amino acid or where R' and R'' together
with the nitrogen and carbon bound to each group respectively form
a pyrrolidinyl group; provided that V and Z are not identical;
provided that when V is C--H, Z is N; T.sup.1 and T.sup.2 are
independently selected from the group consisting of hydrogen,
hydroxyl, C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy,
amino, substituted amino, and halo; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may
be a structure ll ##STR00226## wherein for structure ll: R is
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; Q' is selected from the group
consisting of NH, O, and S; G' is selected from the group
consisting of amino, aminocarbonyl, methylamino, dimethylamino,
acylamino, --SO.sub.3H, --SO.sub.2NH.sub.2, alkoxyamino,
aminocarbonylamino, oxycarbonylamino, HR'NCHR''C(O)NH--, azido,
cyano, halo, hydroxyamino, and hydrazino, where R' is hydrogen and
R'' is a side-chain of an amino acid or where R' and R'' together
with the nitrogen and carbon bound to each group respectively form
a pyrrolidinyl group; Y is selected from the group consisting of a
bond, O, and CH.sub.2; and each of W, W.sup.1, and W.sup.2 is
independently selected from the group consisting of hydrogen,
C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may be a
structure mm ##STR00227## where in for structure mm Q is as defined
for structure kk A and B are independently selected from the group
consisting of C=Q, NH, and methylene optionally substituted with 1
to 2 halo groups, provided that A and B are not both NH; D is NH,
or -D-A-B- together form a --N.dbd.CH--NH--,
--(C=Q)-CH.sub.2--(C=Q)-, --(C=Q)-NH--(C=Q)-,
--(CX')=(CX')--(C=Q)-, or --CH.dbd.CH--NH-- group where X' is halo;
each Q is independently selected from the group consisting of O, S,
and NH; R is selected from the group consisting of hydrogen and
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; T.sup.1 and T.sup.2 are independently
selected from the group consisting of hydrogen, hydroxyl,
C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy, amino,
substituted amino, and halo; Y is selected from the group
consisting of a bond, O, and CH.sub.2; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; and
pharmaceutically acceptable salts, tautomers, pharmaceutically
acceptable salts of tautomers, salts (acidic or basic addition
salts), hydrates, solvates, crystalline forms thereof, optionally
in combination with one or more antiviral, antibacterial, or
antiproliferative agents.
10. A product or composition obtained by a process, which
comprises: (a) deoxygenating the 2'-C-position of a compound
represented by 1 or 1' ##STR00228## to obtain a compound
represented by 2 or 2' ##STR00229## and (b) derivatizing the
compound represented by 2 or 2' to obtain a compound represented by
A or A' ##STR00230## wherein for structures 1, 1', 2, 2', A, and A'
(a) R.sup.2 is independently CH.sub.3, CH.sub.2F, CHF.sub.2,
CF.sub.3, F, CN, C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, or C.sub.1-4
alkyl optionally substituted with amino, hydroxy, or 1 to 3
fluorine atoms; (b) R is H, phosphate, including 5'-monophosphate,
5',3'-cyclic phosphate, diphosphate, triphosphate, or a stabilized
phosphate prodrug, H-phosphonate, including stabilized
H-phosphonates, acyl, including optionally substituted phenyl and
lower acyl, alkyl, including lower alkyl, O-substituted
carboxyalkylamino or its peptide derivatives, sulfonate ester,
including alkyl or arylalkyl sulfonyl, including methanesulfonyl
and benzyl, wherein the phenyl group is optionally substituted, a
lipid, including a phospholipid, an L or D-amino acid, a
carbohydrate, a peptide, a cholesterol, or other pharmaceutically
acceptable leaving group which when administered in vivo; (c)
R.sup.3 is independently OH, H, C.sub.1-4 alkyl, C.sub.2-4 alkenyl,
C.sub.2-4 alkynyl, vinyl, N.sub.3, CN, Cl, Br, F, I, NO.sub.2,
C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.2-4
alkynyl), C(O)O(C.sub.2-4 alkenyl), O(C.sub.1-10 acyl), O(C.sub.1-4
alkyl), O(C.sub.2-4 alkenyl), SH, S(C.sub.1-4 acyl), S(C.sub.1-4
alkyl), S(C.sub.2-4 alkynyl), S(C.sub.2-4 alkenyl), SO(C.sub.1-4
acyl), SO(C.sub.1-4 alkyl), SO(C.sub.2-4 alkynyl), SO(C.sub.2-4
alkenyl), SO.sub.2(C.sub.1-4 acyl), SO.sub.2(C.sub.1-4 alkyl),
SO.sub.2(C.sub.2-4 alkynyl), SO.sub.2(C.sub.2-4 alkenyl),
OS(O).sub.2(C.sub.1-4 acyl), OS(O).sub.2(C.sub.1-4 alkyl),
OS(O).sub.2(C.sub.2-4 alkenyl), NH.sub.2, NH(C.sub.1-4 alkyl),
NH(C.sub.2-4 alkenyl), NH(C.sub.2-4 alkynyl), NH(C.sub.1-4 acyl),
N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-18 acyl).sub.2, wherein alkyl,
alkynyl, alkenyl and vinyl are optionally substituted by N.sub.3,
CN, one to three halogen (Cl, Br, F, I), NO.sub.2, C(O)O(C.sub.1-4
alkyl), C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.2-4 alkynyl),
C(O)O(C.sub.2-4 alkenyl), O(C.sub.1-4 acyl), O(C.sub.1-4 alkyl),
O(C.sub.2-4 alkenyl), SH, S(C.sub.1-4 acyl), S(C.sub.1-4 alkyl),
S(C.sub.2-4 alkynyl), S(C.sub.2-4 alkenyl), SO(C.sub.1-4 acyl),
SO(C.sub.1-4 alkyl), SO(C.sub.2-4 alkynyl), SO(C.sub.2-4 alkenyl),
SO.sub.2(C.sub.1-4 acyl), SO.sub.2(C.sub.1-4 alkyl),
SO.sub.2(C.sub.2-4 alkynyl), SO.sub.2(C.sub.2-4 alkenyl),
OS(O).sub.2(C.sub.1-4 acyl), OS(O).sub.2(C.sub.1-4 alkyl),
OS(O).sub.2(C.sub.2-4 alkenyl), NH.sub.2, NH(C.sub.1-4 alkyl),
NH(C.sub.2-4 alkenyl), NH(C.sub.2-4 alkynyl), NH(C.sub.1-4 acyl),
N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-4 acyl).sub.2; (d) R.sup.4 is
independently H, a lower alkyl, CN, vinyl, O-(lower alkyl),
hydroxyl lower alkyl, i.e., --(CH.sub.2).sub.pOH, where p is 1-6,
including hydroxylmethyl (CH.sub.2OH), CH.sub.2F, N.sub.3,
CH.sub.2CN, CH.sub.2NH.sub.2, CH.sub.2NHCH.sub.3,
CH.sub.2N(CH.sub.3).sub.2, ethynyl alkyne (optionally substituted),
or halogen, including F, Cl, Br, or I, alkenyl, alkynyl, Br-vinyl,
hydroxy, O-alkenyl, NO.sub.2, amino, loweralkylamino, or
di(loweralkyl)amino; (e) R and R.sup.3 can together form
5',3'-cyclic phosphate including stabilized prodrugs thereof, (f)
Base is a naturally occurring or modified purine or pyrimidine base
represented by the following structures: ##STR00231## wherein for a
and b Z is Nor CR.sup.8; R.sup.5, R.sup.6, and R.sup.7 are
independently H, F, Cl, Br, I, OH, OR', SH, SR', NH.sub.2, NHR',
NR'.sub.2 (two R' can form saturated or unsaturated rings, or
saturated or unsaturated heterocyclic rings), lower alkyl of
C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkyl of
C.sub.1-C.sub.6, lower alkenyl of C.sub.2-C.sub.6, halogenated (F,
Cl, Br, I) lower alkenyl of C.sub.2-C.sub.6, lower alkynyl of
C.sub.2-C.sub.6 such as C.ident.CH, halogenated (F, Cl, Br, I)
lower alkynyl of C.sub.2-C.sub.6, lower alkoxy of C.sub.1-C.sub.6,
halogenated (F, Cl, Br, I) lower alkoxy of C.sub.1-C.sub.6,
CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR', CONR.sub.12,
CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R' wherein R' is an
optionally substituted alkyl, which includes, but is not limited
to, an optionally substituted C.sub.1-20 alkyl, an optionally
substituted C.sub.1-10 alkyl, an optionally substituted lower
alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkylaryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; R.sup.8 is
independently H, halogen (including F, Cl, Br, I), OH, OR', SH,
SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form saturated or
unsaturated rings, or saturated or unsaturated heterocyclic rings),
NO.sub.2, lower alkyl of C.sub.1-C.sub.6, halogenated (F, Cl, Br,
I) lower alkyl of C.sub.1-C.sub.6, lower alkenyl of
C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower alkenyl of
C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6, halogenated (F,
Cl, Br, I) lower alkynyl of C.sub.2-C.sub.6, lower alkoxy of
C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkoxy of
C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR',
CONR.sub.12, CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R', wherein R'
is an optionally substituted alkyl, which includes, but is not
limited to, an optionally substituted C.sub.1-20 alkyl, an
optionally substituted C.sub.1-10 alkyl, an optionally substituted
lower alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkyl-aryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; or base may be
selected from a group of formula c ##STR00232## wherein for
structure c, 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, CNNR, 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.sup.X 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; 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; or
base may be a structure selected from the group consisting of
structures d-n ##STR00233## ##STR00234## wherein Z, X, and R'' are
defined as in structure c; base may be a structure selected from
the group consisting of structures o-ff ##STR00235## ##STR00236##
##STR00237## ##STR00238## wherein G and R are defined as in
structure c; base may be a structure gg ##STR00239## 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 defined as in
structure c; base may be a structure hh ##STR00240## 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 in 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 defined as in structure c;
base may be a structure ii ##STR00241## wherein R and G are defined
as in structure c; base may be a structure jj ##STR00242## wherein
R and G are defined as in structure c; or base may be a structure
kk ##STR00243## wherein for structure kk: R is selected from the
group consisting of hydrogen and C.sub.1-C.sub.3 alkyl; X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
Y is selected from the group consisting of a bond, O, and CH.sub.2;
Q is absent or is selected from the group consisting of O, S, and
NH, provided that when Q is absent, V and NH are both attached to a
CH.sub.2 group; V is selected from the group consisting of N and
C-G; Z is selected from the group consisting of N and C-G'; G and
G' are independently selected from the group consisting of
hydrogen, amino, aminocarbonyl, methylamino, dimethylamino,
acylamino, alkoxyamino, --SO.sub.3H, --SO.sub.2NH.sub.2,
aminocarbonylamino, oxycarbonylamino, HR'NCHR''C(O)NH--, azido,
cyano, halo, hydroxyamino, and hydrazino, where R' is hydrogen and
R'' is a side-chain of an amino acid or where R' and R'' together
with the nitrogen and carbon bound to each group respectively form
a pyrrolidinyl group; provided that V and Z are not identical;
provided that when V is C--H, Z is N; T.sup.1 and T.sup.2 are
independently selected from the group consisting of hydrogen,
hydroxyl, C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy,
amino, substituted amino, and halo; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may
be a structure ll ##STR00244## wherein for structure ll: R is
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; Q' is selected from the group
consisting of NH, O, and S; G' is selected from the group
consisting of amino, aminocarbonyl, methylamino, dimethylamino,
acylamino, --SO.sub.3H, --SO.sub.2NH.sub.2, alkoxyamino,
aminocarbonylamino, oxycarbonylamino, HR'NCHR''C(O)NH--, azido,
cyano, halo, hydroxyamino, and hydrazino, where R' is hydrogen and
R'' is a side-chain of an amino acid or where R' and R'' together
with the nitrogen and carbon bound to each group respectively form
a pyrrolidinyl group; Y is selected from the group consisting of a
bond, O, and CH.sub.2; and each of W, W.sup.1, and W.sup.2 is
independently selected from the group consisting of hydrogen,
C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may be a
structure mm ##STR00245## where in for structure mm Q is as defined
for structure kk A and B are independently selected from the group
consisting of C=Q, NH, and methylene optionally substituted with 1
to 2 halo groups, provided that A and B are not both NH; D is NH,
or -D-A-B- together form a --N.dbd.CH--NH--,
--(C=Q)-CH.sub.2--(C=Q)-, --(C=Q)-NH--(C=Q)-,
--(CX')=(CX')--(C=Q)-, or --CH.dbd.CH--NH-- group where X' is halo;
each Q is independently selected from the group consisting of O, S,
and NH; R is selected from the group consisting of hydrogen and
C.sub.1-C.sub.3 alkyl; X is selected from the group consisting of
hydrogen, halo, and OW.sup.2; T.sup.1 and T.sup.2 are independently
selected from the group consisting of hydrogen, hydroxyl,
C.sub.1-C.sub.4-alkoxy, C.sub.1-C.sub.4-thioalkoxy, amino,
substituted amino, and halo; Y is selected from the group
consisting of a bond, O, and CH.sub.2; and each of W, W.sup.1, and
W.sup.2 is independently selected from the group consisting of
hydrogen, C.sub.1-C.sub.4 alkyl, and a prodrug group; and
pharmaceutically acceptable salts, tautomers, pharmaceutically
acceptable salts of tautomers, salts (acidic or basic addition
salts), hydrates, solvates, crystalline forms thereof, optionally
in combination with one or more antiviral, antibacterial, or
antiproliferative agents.
Description
FIELD
[0001] Embodiments of the invention are directed to compounds,
methods, and compositions for use in the treatment of viral
infections. More specifically embodiments of the invention are
2',4'-substituted nucleoside compounds useful for the treatment of
viral infections, such as HIV, HCV, and HBV infections.
BACKGROUND
[0002] Hepatitis C virus (HCV) infection is a major health problem
that leads to chronic liver disease, such as cirrhosis and
hepatocellular carcinoma, in a substantial number of infected
individuals, estimated to be 2-15% of the world's population. There
are an estimated 4.5 million infected people in the United States
alone, according to the U.S. Center for Disease Control. According
to the World Health Organization, there are more than 200 million
infected individuals worldwide, with at least 3 to 4 million people
being infected each year. Once infected, about 20% of people clear
the virus, but the rest can harbor HCV the rest of their lives. Ten
to twenty percent of chronically infected individuals eventually
develop liver-destroying cirrhosis or cancer. The viral disease is
transmitted parenterally by contaminated blood and blood products,
contaminated needles, or sexually and vertically from infected
mothers or carrier mothers to their offspring. Current treatments
for HCV infection, which are restricted to immunotherapy with
recombinant interferon-.alpha. alone or in combination with the
nucleoside analog ribavirin, are of limited clinical benefit as
resistance develops rapidly. Moreover, there is no established
vaccine for HCV. Consequently, there is an urgent need for improved
therapeutic agents that effectively combat chronic HCV
infection.
[0003] The HCV virion is an enveloped positive-strand RNA virus
with a single oligoribonucleotide genomic sequence of about 9600
bases which encodes a polyprotein of about 3,010 amino acids. The
protein products of the HCV gene consist of the structural proteins
C, E1, and E2, and the non-structural proteins NS2, NS3, NS4A and
NS4B, and NS5A and NS5B. The nonstructural (NS) proteins are
believed to provide the catalytic machinery for viral replication.
The NS3 protease releases NS5B, the RNA-dependent RNA polymerase
from the polyprotein chain. HCV NS5B polymerase is required for the
synthesis of a double-stranded RNA from a single-stranded viral RNA
that serves as a template in the replication cycle of HCV.
Therefore, NS5B polymerase is considered to be an essential
component in the HCV replication complex (K. Ishi, et al,
Heptology, 1999, 29: 1227-1235; V. Lohmann, et al., Virology, 1998,
249: 108-118). Inhibition of HCV NS5B polymerase prevents formation
of the double-stranded HCV RNA and therefore constitutes an
attractive approach to the development of HCV-specific antiviral
therapies.
[0004] HCV belongs to a much larger family of viruses that share
many common features.
[0005] Flaviviridae Viruses
[0006] The Flaviviridae family of viruses comprises at least three
distinct genera: pestiviruses, which cause disease in cattle and
pigs; flaviviruses, which are the primary cause of diseases such as
dengue fever and yellow fever; and hepaciviruses, whose sole member
is HCV. The flavivirus genus includes more than 68 members
separated into groups on the basis of serological relatedness
(Calisher et al., J. Gen. Virol, 1993, 70, 37-43). Clinical
symptoms very and include fever, encephalitis and hemorrhagic fever
(Fields Virology, Editors: Fields, B. N., Knipe, D. M., and Howley,
P. M., Lippincott-Raven Publishers, Philadelphia, Pa., 1996,
Chapter 31, 931-959). Flaviviruses of global concern that are
associated with human disease include the Dengue Hemorrhagic Fever
viruses (DHF), yellow fever virus, shock syndrome and Japanese
encephalitis virus (Halstead, S. B., Rev. Infect. Dis., 1984,
6,251-264; Halstead, S. B., Science, 239: 476-481, 1988; Monath, T.
P., New Eng. J. Med. 1988, 319, 641-643).
[0007] The pestivirus genus includes bovine viral diarrhea virus
(BVDV), classical swine fever virus (CSFV, also called hog cholera
virus) and border disease virus (BDV) of sheep (Moennig, V. et al.
Adv. Vir. Res. 1992, 41, 53-98). Pestivirus infections of
domesticated livestock (cattle, pigs, and sheep) cause significant
economic losses worldwide. BVDV causes mucosal disease in cattle
and is of significant economic importance to the livestock industry
(Meyers, G. and Thiel, H. J., Advances in Virus Research, 1996, 47,
53-118; Moennig V., et al., Adv. Vir. Res. 1992, 41, 53-98). Human
pestiviruses have not been as extensively characterized as the
animal pestiviruses. However, serological surveys indicate
considerable pestivirus exposure in humans.
[0008] Pestiviruses and hepaciviruses are closely related virus
groups within the Flaviviridae family. Other closely related
viruses in this family include GB virus A, GB virus A-like agents,
GB virus-B and GB virus-C (also called hepatitis G virus, HGV). The
hepacivirus group (hepatitis C virus; HCV) consists of a number of
closely related but genotypically distinguishable viruses that
infect humans. There are at least 6 HCV genotypes and more than 50
subtypes. Due to the similarities between pestiviruses and
hepaciviruses, combined with the poor ability of hepaciviruses to
grow efficiently in cell culture, bovine viral diarrhea virus
(BVDV) is often used as a surrogate to study the HCV virus.
[0009] The genetic organization of pestiviruses and hepaciviruses
is very similar. These positive stranded RNA viruses possess a
single large open reading frame (ORF) encoding all the viral
proteins necessary for virus replication. These proteins are
expressed as a poly protein that is a co- and post-translationally
processed by both cellular and virus-encoded proteinases to yield
the mature viral proteins. The viral proteins responsible for the
replication of the viral genome RNA are located within
approximately the carboxy-terminal. Two-thirds of the ORF are
termed non (NS) proteins. The genetic organization and polyprotein
processing of the nonstructural protein portion of the ORF for
pestiviruses and hepaciviruses is very similar. For both the
pestiviruses and hepaciviruses, the mature nonstructural (NS
proteins, in sequential order from the amino-terminus of the
nonstructural protein coding region to the carboxy-terminus of the
ORF, consist of p7, NS2, NS3, NS4A, NS4B, NS5A, and NS5B.
[0010] The NS proteins of pestiviruses and hepaciviruses share
sequence domains that are characteristic of specific protein
functions. For example, the NS3 proteins of viruses in both groups
possess amino acid sequence motifs characteristic of serine
proteinases and of helicases (Gorbalenya, et al., Nature, 1988,
333, 22; Bazan and Fletterick, Virology, 1989, 171, 637-639;
Gorbalenyl, et al., Nucleic Acid Res., 1989, 17, 3889-3897).
Similarly, the NS5B proteins of pestiviruses and hepaciviruses have
the motifs characteristic of RNA-directed RNA polymerases (Koonin,
E. V. and Dolja, V. V., Crit. Rev. Biochem. Molec. Biol., 1993, 28,
375-430).
[0011] The actual roles and functions of the NS proteins of
pestiviruses and hepaciviruses in the lifecycle of the viruses are
directly analogous. In both cases, the NS3 serine proteinase is
responsible for all proteolytic processing of polyprotein
precursors down stream of its position in the ORF (Wiskerchen and
Collett, Virology, 1991, 184, 341-350; Bartenschlager et al., J.
Virol. 1993, 67, 3835-3844; Eckart et al., Biochem. Biophys. Res.
Comm. 1993, 192, 399-406; Grakoui et al., J. Virol. 1993, 67,
2832-2843; Grakoui et al., Proc. Natl. Acad. Aci. USA, 1993, 909,
10583-10587; Hijikata et al., J. Virol. 1993, 67, 4665-4675; Tome
et al., J. Virol., 1993, 67, 4017-4026). The NS4A protein, in both
cases, acts as a cofactor with the NS3 serine protease
(Bartenschlager et al., J. Virol. 1994, 68, 5045-5055; failla et
al., J. Virol., 1994, 68, 3753-3760; Xu et al., J. Virol., 1997,
71, 5312-5322). The NS3 protein of both viruses also functions as a
helicase (Kim et al., Biochem Biophys. Res. Comm., 1995, 215,
160-166; Jin and Peterson, Arch. Biochem. Biophys., 1995, 323,
47-53; Warrener and Collett, J. Virol., 1995, 69, 1720-1726).
Finally, the NS5B proteins of pestiviruses and hepaciviruses have
the predicted RNA-directed RNA polymerase activity (Behrens et al.,
EMBO, 1996, 15, 12-22; Lechmann et al., J. Virol., 1997, 71,
8416-8428; Yuan et al., Biochem. Biophys. Res. Comm. 1997, 232,
231-235; Hagedorn PCT WO 97/12033; Zhong et al., J. Virol., 1998,
72, 9365-9369).
[0012] Currently, there are limited treatment options for
individuals infected with hepatitis C virus. The current approved
therapeutic option is the use of immunotherapy with recombinant
interferon-.alpha. alone or in combination with the nucleoside
analog ribavirin. This therapy is limited in its clinical
effectiveness and only 50% of treated patients respond to therapy.
Therefore, there is significant need for more effective and novel
therapies to address the unmet medical need posed by HCV
infection.
[0013] A number of potential molecular targets for drug development
of direct acting antivirals for anti-HCV therapeutics have now been
identified including, but not limited to, the NS2-NS3 autoprotease,
the N3 protease, the N3 helicase and the NS5B polymerase. The
RNA-dependent RNA polymerase is absolutely essential for
replication of the single-stranded, positive sense, RNA genome and
this enzyme has elicited significant interest among medicinal
chemists.
[0014] Inhibitors of HCV NS5B as potential therapies for HCV
infection have been reviewed: Tan, S. L., et al., Nature Rev. Drug
Discov., 2002, 1, 867-881; Walker, M. P. et al., Exp. Opin.
Investigational Drugs, 2003, 12, 1269-1280; Ni, Z-J., et al.,
Current Opinion in Drug Discovery and Development, 2004, 7,
446-459; Beaulieu, P. L., et al., Current Opinion in
Investigational Drugs, 2004, 5, 838-850; Wu, J., et al., Current
Drug Targets-Infectious Disorders, 2003, 3, 207-219; Griffith, R.
C., et al., Annual Reports in Medicinal Chemistry, 2004, 39,
223-237; Carrol, S., et al., Infectious Disorders-Drug Targets,
20-06, 6, 17-29. The potential for the emergence of resistant HCV
strains and the need to identify agents with broad genotype
coverage supports the need for continuing efforts to identify novel
and more effective nucleosides as HCV NS5B inhibitors. Nucleoside
inhibitors of NS5b polymerase can act either as a non-natural
substrate that results in chain termination or as a competitive
inhibitor which competes with nucleotide binding to the polymerase.
To function as a chain terminator the nucleoside analog must be
taken up by the cell and converted in vivo to a triphosphate to
compete for the polymerase nucleotide binding site. This conversion
to the triphosphate is commonly mediated by cellular kinases which
imparts additional structural requirements on a potential
nucleoside polymerase inhibitor.
[0015] Despite the existence of efficient vaccines, hepatitis B
virus (HBV) infection remains a major public health problem
worldwide with 400 million chronic carriers. These infected
patients are exposed to a risk of developing liver cirrhosis and
hepatocellular carcinoma (Lee, W. M. 1997, N. Eng. J. Med., 337,
1733-1745). Currently, there are believed to be approximately 1.25
million chronic hepatitis B carriers just in the United States,
with 200,000 people newly infected each year by contact with blood
or body fluids.
[0016] Hepatitis B virus is second to tobacco as a cause of human
cancer. The mechanism by which HBV induces cancer is unknown,
although it is postulated that may directly trigger tumor
development, or indirectly trigger tumor development through
chronic inflammation, cirrhosis, and cell regeneration associated
with the infection.
[0017] Hepatitis B virus has reached epidemic levels worldwide.
After a two to six month incubation period in which the host is
unaware of the infection, HBV infection can lead to acute hepatitis
and liver damage, that causes abdominal pain, jaundice, and
elevated blood levels of certain enzymes. HBV can cause fulminant
hepatitis, a rapidly progressive, often fatal form of the disease
in which massive sections of the liver are destroyed. Patients
typically recover from acute viral hepatitis. In some patients,
however, high levels of viral antigen persist in the blood for an
extended, or indefinite, period, causing a chronic infection.
Chronic infections can lead to chronic persistent hepatitis.
Patients infected with chronic persistent HBV are most common in
developing countries. By mid-1991, there were approximately 225
million chronic carriers of HBV in Asia alone, and worldwide,
almost 300 million carriers. Chronic persistent hepatitis can cause
fatigue, cirrhosis of the liver, and hepatocellular carcinoma, a
primary liver cancer.
[0018] In western industrialized countries, high risk groups for
HBV infection include those in contact with HBV carriers or their
blood samples. The epidemiology of HBV is in fact very similar to
that of HIV, which accounts for why HBV infection is common among
patients with AIDS or HIV-associated infections. However, HBV is
more contagious than HIV.
[0019] As is well known, acquired immune deficiency syndrome (AIDS)
is a disease that severely compromises the human immune system, and
that leads to death. The cause of AIDS has been determined to be
the human immunodeficiency virus (HIV). To ameliorate suffering and
to prolong the lives of infected hosts new compounds and methods of
treating AIDS and attacking the HIV virus continue to be
sought.
[0020] The preceding references and all other references cited in
the present specification are hereby incorporated herein by
reference.
SUMMARY
[0021] Embodiments of the invention are directed to novel
2',4'-substituted nucleoside derivatives for the treatment of viral
infections in mammals. Thus, in one aspect, an antivirally
effective nucleoside is a 2',4'-disubstituted 2'-deoxynucleoside
(.beta.-D or .beta.-L), its 5'-mono-phosphate, its 5',3'-cyclic
phosphate, its 5'-diphosphate and its 5'-triphosphate or its
pharmaceutically acceptable salt (acidic or basic addition salt),
hydrate, solvate, crystalline form or prodrug thereof of the
general formula:
##STR00001## [0022] wherein
[0023] (a) R.sup.2 is independently CH.sub.3, CH.sub.2F, CHF.sub.2,
CF.sub.3, F, CN, C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, or C.sub.1-4
alkyl optionally substituted with amino, hydroxy, or 1 to 3
fluorine atoms;
[0024] (b) R is H, phosphate, including 5'-monophosphate,
5',3'-cyclic phosphate, diphosphate, triphosphate, or a stabilized
phosphate prodrug, H-phosphonate, including stabilized
H-phosphonates, acyl, including optionally substituted phenyl and
lower acyl, alkyl, including lower alkyl, O-substituted
carboxyalkylamino or its peptide derivatives, sulfonate ester,
including alkyl or arylalkyl sulfonyl, including methanesulfonyl
and benzyl, wherein the phenyl group is optionally substituted, a
lipid, including a phospholipid, an L or D-amino acid, a
carbohydrate, a peptide, a cholesterol, or other pharmaceutically
acceptable leaving group which when administered in vivo;
[0025] (c) R.sup.3 is independently OH, H, C.sub.1-4 alkyl,
C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, vinyl, N.sub.3, CN, Cl, Br,
F, I, NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4 alkyl),
C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl), O(C.sub.1-10
acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH, S(C.sub.1-4
acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl), S(C.sub.2-4
alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl), SO(C.sub.2-4
alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4 acyl),
SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.1-4 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2,
N(C.sub.1-18 acyl).sub.2, wherein alkyl, alkynyl, alkenyl and vinyl
are optionally substituted by N.sub.3, CN, one to three halogen
(Cl, Br, F, I), NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4
alkyl), C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl),
O(C.sub.1-4 acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH,
S(C.sub.1-4 acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl),
S(C.sub.2-4 alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl),
SO(C.sub.2-4 alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4
acyl), SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.1-4 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-4
acyl).sub.2;
[0026] (d) R.sup.4 is independently H, a lower alkyl, CN, vinyl,
O-(lower alkyl), hydroxyl lower alkyl, i.e., --(CH.sub.2).sub.pOH,
where p is 1-6, including hydroxylmethyl (CH.sub.2OH), CH.sub.2F,
N.sub.3, CH.sub.2CN, CH.sub.2NH.sub.2, CH.sub.2NHCH.sub.3,
CH.sub.2N(CH.sub.3).sub.2, ethynyl alkyne (optionally substituted),
or halogen, including F, Cl, Br, or I, alkenyl, alkynyl, Br-vinyl,
hydroxy, O-alkenyl, NO.sub.2, amino, loweralkylamino, or
di(loweralkyl)amino;
[0027] (e) R and R.sup.3 can together form 5',3'-cyclic phosphate
including stabilized prodrugs thereof;
[0028] (f) Base (B) is a naturally occurring or modified purine or
pyrimidine base represented by the following structures:
##STR00002##
[0029] wherein for a and b
[0030] Z is N or CR.sup.8;
[0031] R.sup.5, R.sup.6, and R.sup.7 are independently H, F, Cl,
Br, I, OH, OR', SH, SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form
saturated or unsaturated rings, or saturated or unsaturated
heterocyclic rings), lower alkyl of C.sub.1-C.sub.6, halogenated
(F, Cl, Br, I) lower alkyl of C.sub.1-C.sub.6, lower alkenyl of
C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower alkenyl of
C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6 such as
C.ident.CH, halogenated (F, Cl, Br, I) lower alkynyl of
C.sub.2-C.sub.6, lower alkoxy of C.sub.1-C.sub.6, halogenated (F,
Cl, Br, I) lower alkoxy of C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R',
CONH.sub.2, CONHR', CONR.sub.12, CH.dbd.CHCO.sub.2H, or
CH.dbd.CHCO.sub.2R', wherein R' is an optionally substituted alkyl,
which includes, but is not limited to, an optionally substituted
C.sub.1-20 alkyl, an optionally substituted C.sub.1-10 alkyl, an
optionally substituted lower alkyl; an optionally substituted
cycloalkyl; an optionally substituted alkynyl of C.sub.2-C.sub.6,
an optionally substituted lower alkenyl of C.sub.2-C.sub.6, or
optionally substituted acyl, which includes but is not limited to
C(O)alkyl, C(O)(C.sub.1-20 alkyl), C(O)(C.sub.1-10 alkyl), or
C(O)(lower alkyl), optionally substituted aryl, optionally
substituted C.sub.1-C.sub.4 alkyl-aryloxy, heteroaryl, optionally
substituted C.sub.1-C.sub.4 alkyl-heteroaryl, an optionally
substituted alkoxy C.sub.1-20 alkyl, an optionally substituted
amino C.sub.1-20 alkyl, an optionally substituted fluoro C.sub.1-20
alkyl;
[0032] R.sup.8 is independently H, halogen (including F, Cl, Br,
I), OH, OR', SH, SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form
saturated or unsaturated rings, or saturated or unsaturated
heterocyclic rings), NO.sub.2, lower alkyl of C.sub.1-C.sub.6,
halogenated (F, Cl, Br, I) lower alkyl of C.sub.1-C.sub.6, lower
alkenyl of C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower
alkenyl of C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6,
halogenated (F, Cl, Br, I) lower alkynyl of C.sub.2-C.sub.6, lower
alkoxy of C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkoxy
of C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR',
CONR.sub.12, CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R' wherein R'
is an optionally substituted alkyl, which includes, but is not
limited to, an optionally substituted C.sub.1-20 alkyl, an
optionally substituted C.sub.1-10 alkyl, an optionally substituted
lower alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkyl-aryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; or
base (B) may be selected from a group of formula c
##STR00003##
wherein for structure c, 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, CNNR, 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 linesindicate a possible pi or double
bond; 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; or base (B) may be a structure
selected from the group consisting of structures d-n
##STR00004## ##STR00005##
wherein Z, X, and R'' are defined as in structure c; base may be a
structure selected from the group consisting of structures o-ff
##STR00006## ##STR00007## ##STR00008##
wherein G and R are defined as in structure c; base (B) may be a
structure gg
##STR00009##
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
defined as in structure c; base (B) may be a structure hh
##STR00010##
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 in
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 defined
as in structure c; base (B) may be a structure ii
##STR00011##
wherein R and G are defined as in structure c; base may be a
structure jj
##STR00012##
wherein R and G are defined as in structure c; or base may be a
structure kk
##STR00013##
wherein for structure kk: [0033] R is selected from the group
consisting of hydrogen and C.sub.1-C.sub.3 alkyl; [0034] X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
[0035] Y is selected from the group consisting of a bond, O, and
CH.sub.2; [0036] Q is absent or is selected from the group
consisting of O, S, and NH, provided that when Q is absent, V and
NH are both attached to a CH.sub.2 group; [0037] V is selected from
the group consisting of N and C-G; [0038] Z is selected from the
group consisting of N and C-G'; [0039] G and G' are independently
selected from the group consisting of hydrogen, amino,
aminocarbonyl, methylamino, dimethylamino, acylamino, alkoxyamino,
--SO.sub.3H, --SO.sub.2NH.sub.2, aminocarbonylamino,
oxycarbonylamino, HR'NCHR''C(O)NH--, azido, cyano, halo,
hydroxyamino, and hydrazino, where R' is hydrogen and R'' is a
side-chain of an amino acid or where R' and R'' together with the
nitrogen and carbon bound to each group respectively form a
pyrrolidinyl group; [0040] provided that V and Z are not identical;
[0041] provided that when V is C--H, Z is N; [0042] T.sup.1 and
T.sup.2 are independently selected from the group consisting of
hydrogen, hydroxyl, C.sub.1-C.sub.4-alkoxy,
C.sub.1-C.sub.4-thioalkoxy, amino, substituted amino, and halo; and
[0043] each of W, W.sup.1, and W.sup.2 is independently selected
from the group consisting of hydrogen, C.sub.1-C.sub.4 alkyl, and a
prodrug group; or [0044] base may be a structure ll
##STR00014##
[0044] wherein for structure ll: [0045] R is C.sub.1-C.sub.3 alkyl;
[0046] X is selected from the group consisting of hydrogen, halo,
and OW.sup.2; [0047] Q' is selected from the group consisting of
NH, O, and S; [0048] G' is selected from the group consisting of
amino, aminocarbonyl, methylamino, dimethylamino, acylamino,
--SO.sub.3H, --SO.sub.2NH.sub.2, alkoxyamino, aminocarbonylamino,
oxycarbonylamino, HR'NCHR''C(O)NH--, azido, cyano, halo,
hydroxyamino, and hydrazino, where R' is hydrogen and R'' is a
side-chain of an amino acid or where R' and R'' together with the
nitrogen and carbon bound to each group respectively form a
pyrrolidinyl group; Y is selected from the group consisting of a
bond, O, and CH.sub.2; and each of W, W.sup.1, and W.sup.2 is
independently selected from the group consisting of hydrogen,
C.sub.1-C.sub.4 alkyl, and a prodrug group; or [0049] base may be a
structure mm
##STR00015##
[0049] where in for structure mm [0050] Q is as defined for
structure kk [0051] A and B are independently selected from the
group consisting of C=Q, NH, and methylene optionally substituted
with 1 to 2 halo groups, provided that A and B are not both NH;
[0052] D is NH, or -D-A-B- together form a --N.dbd.CH--NH--,
--(C=Q)-CH.sub.2--(C=Q)-, --(C=Q)-NH--(C=Q)-,
--(CX')=(CX')--(C=Q)-, or --CH.dbd.CH--NH-- group where X' is halo;
[0053] each Q is independently selected from the group consisting
of O, S, and NH; R is selected from the group consisting of
hydrogen and C.sub.1-C.sub.3 alkyl; [0054] X is selected from the
group consisting of hydrogen, halo, and OW.sup.2; [0055] T.sup.1
and T.sup.2 are independently selected from the group consisting of
hydrogen, hydroxyl, C.sub.1-C.sub.4-alkoxy,
C.sub.1-C.sub.4-thioalkoxy, amino, substituted amino, and halo;
[0056] Y is selected from the group consisting of a bond, O, and
CH.sub.2; and each of W, W.sup.1, and W.sup.2 is independently
selected from the group consisting of hydrogen, C.sub.1-C.sub.4
alkyl, and a prodrug group; and pharmaceutically acceptable salts,
tautomers, pharmaceutically acceptable salts of tautomers, salts
(acidic or basic addition salts), hydrates, solvates, crystalline
forms thereof, optionally in combination with one or more
antiviral, antibacterial, or antiproliferative agents.
[0057] It is an object of embodiments of the invention to provide a
compound, method, and composition for the treatment or prevention
of HIV, HBV, or HCV infection in a host. It is a further object of
embodiments of the invention to provide a compound, method, and
composition for the treatment or prevention of HIV, HBV, or HCV
when the host is a human, or when the host is an animal.
DETAILED DESCRIPTION
[0058] Embodiments of the invention are directed to novel
2',4'-substituted nucleoside derivatives for the treatment is viral
infections in mammals, comprised of one or more compounds of the
formula:
##STR00016## [0059] wherein
[0060] (a) R.sup.2 is independently CH.sub.3, CH.sub.2F, CHF.sub.2,
CF.sub.3, F, CN, C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, or C.sub.1-4
alkyl optionally substituted with amino, hydroxy, or 1 to 3
fluorine atoms;
[0061] (b) R is H, phosphate, including 5'-monophosphate,
5',3'-cyclic phosphate, diphosphate, triphosphate, or a stabilized
phosphate prodrug, H-phosphonate, including stabilized
H-phosphonates, acyl, including optionally substituted phenyl and
lower acyl, alkyl, including lower alkyl, O-substituted
carboxyalkylamino or its peptide derivatives, sulfonate ester,
including alkyl or arylalkyl sulfonyl, including methanesulfonyl
and benzyl, wherein the phenyl group is optionally substituted, a
lipid, including a phospholipid, an L or D-amino acid, a
carbohydrate, a peptide, a cholesterol, or other pharmaceutically
acceptable leaving group which when administered in vivo;
[0062] (c) R.sup.3 is independently OH, H, C.sub.1-4 alkyl,
C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, vinyl, N.sub.3, CN, Cl, Br,
F, I, NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4 alkyl),
C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl), O(C.sub.1-10
acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH, S(C.sub.1-4
acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl), S(C.sub.2-4
alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl), SO(C.sub.2-4
alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4 acyl),
SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.1-4 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2,
N(C.sub.1-18 acyl).sub.2, wherein alkyl, alkynyl, alkenyl and vinyl
are optionally substituted by N.sub.3, CN, one to three halogen
(Cl, Br, F, I), NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4
alkyl), C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl),
O(C.sub.1-4 acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH,
S(C.sub.1-4 acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl),
S(C.sub.2-4 alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl),
SO(C.sub.2-4 alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4
acyl), SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.1-4 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-4
acyl).sub.2;
[0063] (d) R.sup.4 is independently H, a lower alkyl, CN, vinyl,
O-(lower alkyl), hydroxyl lower alkyl, i.e., --(CH.sub.2).sub.pOH,
where p is 1-6, including hydroxylmethyl (CH.sub.2OH), CH.sub.2F,
N.sub.3, CH.sub.2CN, CH.sub.2NH.sub.2, CH.sub.2NHCH.sub.3,
CH.sub.2N(CH.sub.3).sub.2, ethynyl alkyne (optionally substituted),
or halogen, including F, Cl, Br, or I, alkenyl, alkynyl, Br-vinyl,
hydroxy, O-alkenyl, NO.sub.2, amino, loweralkylamino, or
di(loweralkyl)amino;
[0064] (e) R and R.sup.3 can together form 5',3'-cyclic phosphate
including stabilized prodrugs thereof;
[0065] (f) Base is a naturally occurring or modified purine or
pyrimidine base represented by the following structures:
##STR00017##
[0066] wherein for a and b
[0067] Z is N or CR.sup.8;
[0068] R.sup.5, R.sup.6, and R.sup.7 are independently H, F, Cl,
Br, I, OH, OR', SH, SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form
saturated or unsaturated rings, or saturated or unsaturated
heterocyclic rings), lower alkyl of C.sub.1-C.sub.6, halogenated
(F, Cl, Br, I) lower alkyl of C.sub.1-C.sub.6, lower alkenyl of
C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower alkenyl of
C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6 such as
C.ident.CH, halogenated (F, Cl, Br, I) lower alkynyl of
C.sub.2-C.sub.6, lower alkoxy of C.sub.1-C.sub.6, halogenated (F,
Cl, Br, I) lower alkoxy of C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R',
CONH.sub.2, CONHR', CONR.sub.12, CH.dbd.CHCO.sub.2H, or
CH.dbd.CHCO.sub.2R' wherein R' is an optionally substituted alkyl,
which includes, but is not limited to, an optionally substituted
C.sub.1-20 alkyl, an optionally substituted C.sub.1-10 alkyl, an
optionally substituted lower alkyl; an optionally substituted
cycloalkyl; an optionally substituted alkynyl of C.sub.2-C.sub.6,
an optionally substituted lower alkenyl of C.sub.2-C.sub.6, or
optionally substituted acyl, which includes but is not limited to
C(O) alkyl, C(O)(C.sub.1-20 alkyl), C(O)(C.sub.1-10 alkyl), or
C(O)(lower alkyl), optionally substituted aryl, optionally
substituted C.sub.1-C.sub.4 alkyl-aryloxy, heteroaryl, optionally
substituted C.sub.1-C.sub.4 alkyl-heteroaryl, an optionally
substituted alkoxy C.sub.1-20 alkyl, an optionally substituted
amino C.sub.1-20 alkyl, an optionally substituted fluoro C.sub.1-20
alkyl;
[0069] R.sup.8 is independently H, halogen (including F, Cl, Br,
I), OH, OR', SH, SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form
saturated or unsaturated rings, or saturated or unsaturated
heterocyclic rings), NO.sub.2, lower alkyl of C.sub.1-C.sub.6,
halogenated (F, Cl, Br, I) lower alkyl of C.sub.1-C.sub.6, lower
alkenyl of C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower
alkenyl of C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6,
halogenated (F, Cl, Br, I) lower alkynyl of C.sub.2-C.sub.6, lower
alkoxy of C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkoxy
of C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR',
CONR.sub.12, CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R', wherein R'
is an optionally substituted alkyl, which includes, but is not
limited to, an optionally substituted C.sub.1-20 alkyl, an
optionally substituted C.sub.1-10 alkyl, an optionally substituted
lower alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkyl-aryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; or
base may be selected from a group of formula c
##STR00018##
wherein for structure c, 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;
[0070] 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, CNNR, SO, S(O).sub.2, SeO and Se(O) 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.sup.X 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 linesindicate a possible pi or double bond;
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; or base may be a structure
selected from the group consisting of structures d-n
##STR00019## ##STR00020##
wherein Z, X, and R'' are defined as in structure c; base may be a
structure selected from the group consisting of structures o-ff
##STR00021## ##STR00022## ##STR00023##
wherein G and R are defined as in structure c; base may be a
structure gg
##STR00024##
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
defined as in structure c; base may be a structure hh
##STR00025##
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 in
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 defined
as in structure c; base may be a structure ii
##STR00026##
wherein R and G are defined as in structure c; base may be a
structure jj
##STR00027##
wherein R and G are defined as in structure c; or base may be a
structure kk
##STR00028##
wherein for structure kk: [0071] R is selected from the group
consisting of hydrogen and C.sub.1-C.sub.3 alkyl; [0072] X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
[0073] Y is selected from the group consisting of a bond, O, and
CH.sub.2; [0074] Q is absent or is selected from the group
consisting of O, S, and NH, provided that when Q is absent, V and
NH are both attached to a CH.sub.2 group; [0075] V is selected from
the group consisting of N and C-G; [0076] Z is selected from the
group consisting of N and C-G'; [0077] G and G' are independently
selected from the group consisting of hydrogen, amino,
aminocarbonyl, methylamino, dimethylamino, acylamino, alkoxyamino,
--SO.sub.3H, --SO.sub.2NH.sub.2, aminocarbonylamino,
oxycarbonylamino, HR'NCHR''C(O)NH--, azido, cyano, halo,
hydroxyamino, and hydrazino, where R' is hydrogen and R'' is a
side-chain of an amino acid or where R' and R'' together with the
nitrogen and carbon bound to each group respectively form a
pyrrolidinyl group; [0078] provided that V and Z are not identical;
[0079] provided that when V is C--H, Z is N; [0080] T.sup.1 and
T.sup.2 are independently selected from the group consisting of
hydrogen, hydroxyl, C.sub.1-C.sub.4-alkoxy,
C.sub.1-C.sub.4-thioalkoxy, amino, substituted amino, and halo; and
[0081] each of W, W.sup.1, and W.sup.2 is independently selected
from the group consisting of hydrogen, C.sub.1-C.sub.4 alkyl, and a
prodrug group; or base may be a structure ll
##STR00029##
[0081] wherein for structure ll: R is C.sub.1-C.sub.3 alkyl; X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
Q' is selected from the group consisting of NH, O, and S; G' is
selected from the group consisting of amino, aminocarbonyl,
methylamino, dimethylamino, acylamino, --SO.sub.3H,
--SO.sub.2NH.sub.2, alkoxyamino, aminocarbonylamino,
oxycarbonylamino, HR'NCHR''C(O)NH--, azido, cyano, halo,
hydroxyamino, and hydrazino, where R' is hydrogen and R'' is a
side-chain of an amino acid or where R' and R'' together with the
nitrogen and carbon bound to each group respectively form a
pyrrolidinyl group; Y is selected from the group consisting of a
bond, O, and CH.sub.2; and each of W, W.sup.1, and W.sup.2 is
independently selected from the group consisting of hydrogen,
C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may be a
structure mm
##STR00030##
where in for structure mm [0082] Q is as defined for structure kk
[0083] A and B are independently selected from the group consisting
of C=Q, NH, and methylene optionally substituted with 1 to 2 halo
groups, provided that A and B are not both NH; [0084] D is NH, or
-D-A-B- together form a --N.dbd.CH--NH--, --(C=Q)-CH.sub.2--(C=Q)-,
--(C=Q)-NH--(C=Q)-, --(CX')=(CX')--(C=Q)-, or --CH.dbd.CH--NH--
group where X' is halo; [0085] each Q is independently selected
from the group consisting of O, S, and NH; R is selected from the
group consisting of hydrogen and C.sub.1-C.sub.3 alkyl; [0086] X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
[0087] T.sup.1 and T.sup.2 are independently selected from the
group consisting of hydrogen, hydroxyl, C.sub.1-C.sub.4-alkoxy,
C.sub.1-C.sub.4-thioalkoxy, amino, substituted amino, and halo;
[0088] Y is selected from the group consisting of a bond, O, and
CH.sub.2; and each of W, W.sup.1, and W.sup.2 is independently
selected from the group consisting of hydrogen, C.sub.1-C.sub.4
alkyl, and a prodrug group; and pharmaceutically acceptable salts,
tautomers, pharmaceutically acceptable salts of tautomers, salts
(acidic or basic addition salts), hydrates, solvates, crystalline
forms thereof, optionally in combination with one or more
antiviral, antibacterial, or antiproliferative agents.
[0089] Particularly preferred embodiments of the invention are
listed, but not limited, in Tables I and II below.
TABLE-US-00001 TABLE I ##STR00031## R.sup.2 R.sup.3 R.sup.4 R.sup.5
X R.sup.6 CH.sub.3 OH N.sub.3 NH.sub.2, NH--NH.sub.2, NH--OH, OH,
SH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2 CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH
CH.sub.3 OH C.ident.CH NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S
H, CH.sub.3, F, Cl, Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr,
CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH CH.sub.3 OH
CH.dbd.CH.sub.2 NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr,
CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH CH.sub.3 OH CN
NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl,
Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl,
C.ident.CH, CN, NH.sub.2, CH.sub.2OH CH.sub.3 OH OMe NH.sub.2,
NH--NH.sub.2, NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl, Br, I,
NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH,
CN, NH.sub.2, CH.sub.2OH CH.sub.3 OH F NH.sub.2, NH--NH.sub.2,
NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2,
CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN,
NH.sub.2, CH.sub.2OH CH.sub.3 OH Me NH.sub.2, NH--NH.sub.2, NH--OH,
OH, SH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH
CH.dbd.CH.sub.2 OH C.ident.CH NH.sub.2, NH--NH.sub.2, NH--OH, OH,
SH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH
CH.dbd.CH.sub.2 OH CN NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S
H, CH.sub.3, F, Cl, Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr,
CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH CH.dbd.CH.sub.2
OH F NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S H, CH.sub.3, F,
Cl, Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl,
C.ident.CH, CN, NH.sub.2, CH.sub.2OH CH.dbd.CH.sub.2 OH Me
NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl,
Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl,
C.ident.CH, CN, NH.sub.2, CH.sub.2OH CH.dbd.CH.sub.2 OH N.sub.3
NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl,
Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl,
C.ident.CH, CN, NH.sub.2, CH.sub.2OH C.ident.CH OH C.ident.CH
NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl,
Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl,
C.ident.CH, CN, NH.sub.2, CH.sub.2OH C.ident.CH OH CN NH.sub.2,
NH--NH.sub.2, NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl, Br, I,
NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH,
CN, NH.sub.2, CH.sub.2OH C.ident.CH OH N.sub.3 NH.sub.2,
NH--NH.sub.2, NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl, Br, I,
NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH,
CN, NH.sub.2, CH.sub.2OH C.ident.CH OH OMe NH.sub.2, NH--NH.sub.2,
NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2
CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN,
NH.sub.2, CH.sub.2OH CH.sub.2F OH C.ident.CH NH.sub.2,
NH--NH.sub.2, NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl, Br, I,
NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH,
CN, NH.sub.2, CH.sub.2OH CH.sub.2F OH CN NH.sub.2, NH--NH.sub.2,
NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2,
CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN,
NH.sub.2, CH.sub.2OH CH.sub.2F OH N.sub.3 NH.sub.2, NH--NH.sub.2,
NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2,
CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN,
NH.sub.2, CH.sub.2OH CH.sub.2F OH CH.dbd.CH.sub.2 NH.sub.2,
NH--NH.sub.2, NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl, Br, I,
NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH,
CN, NH.sub.2, CH.sub.2OH CN OH C.ident.CH NH.sub.2, NH--NH.sub.2,
NH--OH, OH, SH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2,
CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN,
NH.sub.2, CH.sub.2OH CN OH CN NH.sub.2, NH--NH.sub.2, NH--OH, OH,
SH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH CN
OH N.sub.3 NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr,
CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH CN OH
CH.dbd.CH.sub.2 NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr,
CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH CH.sub.2CH.sub.3
OH N.sub.3 NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr,
CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH CF.sub.3 OH
N.sub.3 NH.sub.2, NH--NH.sub.2, NH--OH, OH, SH, O, S H, CH.sub.3,
F, Cl, Br, I, NO.sub.2, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl,
C.ident.CH, CN, NH.sub.2, CH.sub.2OH
[0090] In which a more preferred embodiment is one that has the
following structure, but the same substituent pattern as noted in
Table I.
TABLE-US-00002 TABLE II ##STR00032## ##STR00033## R.sup.2 R.sup.3
R.sup.4 R.sup.6 R.sup.5 Z R.sup.8 R.sup.7 CH.sub.3 OH N.sub.3 H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.3 OH N.sub.3 H NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.sub.3 OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, N -- H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH
N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C
H, F, Cl, Br, I, H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe CH.sub.3 OH C.ident.CH H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.3 OH C.ident.CH H NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.sub.3 OH C.ident.CH NH.sub.2 NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, N -- H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH
C.ident.CH NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle,
C H, F, Cl, Br, I, H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe CH.sub.3 OH
CH.dbd.CH.sub.2 H NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N
-- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt,
OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH CH.dbd.CH.sub.2 H NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CH.sub.3 OH CH.dbd.CH.sub.2
NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.3 OH CH.dbd.CH.sub.2 NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CH.sub.3 OH CN H NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH
CN H NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl,
Br, I, H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CH.sub.3 OH CN NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.sub.3 OH CN NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe CH.sub.3
OH OMe H NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.3 OH OMe H NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.sub.3 OH OMe NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH OMe NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CH.sub.3 OH F H NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH
F H NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br,
I, H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CH.sub.3 OH F NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.sub.3 OH F NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe CH.sub.3
OH Me H NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.3 OH Me H NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.sub.3 OH Me NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH Me NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CH.dbd.CH.sub.2 OH C.ident.CH H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH C.ident.CH H NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.dbd.CH.sub.2 OH C.ident.CH NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl
CH.dbd.CH.sub.2 OH C.ident.CH NH.sub.2 NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.dbd.CH.sub.2 OH CN H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH CN H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, COOMe CH.dbd.CH.sub.2 OH CN NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.dbd.CH.sub.2 OH CN NH.sub.2 NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.dbd.CH.sub.2 OH F H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH F H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe
CH.dbd.CH.sub.2 OH F NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH F NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CH.dbd.CH.sub.2 OH Me H NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.dbd.CH.sub.2 OH Me H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe
CH.dbd.CH.sub.2 OH Me NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH Me
NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F,
Cl, Br, I, H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe CH.dbd.CH.sub.2 OH
N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH N.sub.3 H NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.dbd.CH.sub.2 OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, N -- H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl
CH.dbd.CH.sub.2 OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe C.ident.CH OH C.ident.CH H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH C.ident.CH H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe C.ident.CH OH C.ident.CH NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl C.ident.CH OH C.ident.CH NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe C.ident.CH OH CN H NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl C.ident.CH OH CN H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe C.ident.CH
OH CN NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N --
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl C.ident.CH OH CN NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe C.ident.CH OH N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH N.sub.3 H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe C.ident.CH OH N.sub.3 NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl C.ident.CH OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe C.ident.CH OH OMe H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH OMe H NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe C.ident.CH OH OMe NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl C.ident.CH OH OMe NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.sub.2F OH C.ident.CH H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH C.ident.CH H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CH.sub.2F OH C.ident.CH NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.2F OH C.ident.CH NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CH.sub.2F OH CN H NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F
OH CN H NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl,
Br, I, H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CH.sub.2F OH CN NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.sub.2F OH CN NH.sub.2 NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.sub.2F OH N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH N.sub.3 H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CH.sub.2F OH N.sub.3 NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.2F OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.sub.2F OH CH.dbd.CH.sub.2 H NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, N -- H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F
OH CH.dbd.CH.sub.2 H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe CH.sub.2F
OH CH.dbd.CH.sub.2 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH CH.dbd.CH.sub.2
NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F,
Cl, Br, I, H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe CN OH C.ident.CH H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CN OH C.ident.CH H NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CN OH C.ident.CH NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH C.ident.CH NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CN OH CN H NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, N -- H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH CN H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CN OH CN NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH CN
NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F,
Cl, Br, I, H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe CN OH N.sub.3 H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CN OH N.sub.3 H NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CN OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH N.sub.3 NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CN OH CH.dbd.CH.sub.2 H NH.sub.2,
NH-Me, NH-Et, NH-propyl, heterocycle, N -- H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CN OH CH.dbd.CH.sub.2 H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe CN OH
CH.dbd.CH.sub.2 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH CH.dbd.CH.sub.2
NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F,
Cl, Br, I, H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
NO.sub.2, CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl COOH, CONH.sub.2, COOMe CH.sub.2CH.sub.3 OH
N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.2CH.sub.3 OH N.sub.3 H NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CH.sub.2CH.sub.3 OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, N -- H NH-cyclopropyl, NH-acetyl,
NH-cyclobutyl, NH- t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl
CH.sub.2CH.sub.3 OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et,
NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe CF.sub.3 OH N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH-propyl,
heterocycle, N -- H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH-
t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH,
OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CF.sub.3 OH N.sub.3 H
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I,
H NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- NO.sub.2,
CH.dbd.CH.sub.2, t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, C.ident.CH, CN, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl COOH, CONH.sub.2, COOMe CF.sub.3 OH N.sub.3 NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH-propyl, heterocycle, N -- H
NH-cyclopropyl, NH-acetyl, NH-cyclobutyl, NH- t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CF.sub.3 OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH-propyl, heterocycle, C H, F, Cl, Br, I, H NH-cyclopropyl,
NH-acetyl, NH-cyclobutyl, NH- NO.sub.2, CH.dbd.CH.sub.2, t-butyl,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, C.ident.CH,
CN, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl COOH, CONH.sub.2,
COOMe
[0091] In which a more preferred embodiment is one that has the
following structure, but the same substituent pattern as noted in
Table II.
##STR00034##
DEFINITIONS
[0092] The phrase "a" or "an" entity as used herein refers to one
or more of that entity; for example, a compound refers to one or
more compounds or at least one compound. As such, the terms "a" or
(or "an"), "one or more", and "at least one" can be used
interchangeably herein.
[0093] The phrase "as defined herein above" refers to the first
definition provided in the Summary of the Invention.
[0094] The terms "optional" or "optionally" as used herein means
that a subsequently described event or circumstance may but need
not occur, and that the description includes instances where the
event or circumstance occurs and instances in which it does not.
For example, "optional bond" means that the bond may or may not be
present, and that the description includes single, double, or
triple bonds.
[0095] The term "independently" is used herein to indicate that a
variable is applied in any one instance without regard to the
presence or absence of a variable having that same or a different
definition within the same compound. Thus, in a compound in which R
appears twice and is defined as "independently carbon or nitrogen",
both R's can be carbon, both R's can be nitrogen, or one R can be
carbon and the other nitrogen.
[0096] The term "alkenyl" refers to an unsubstituted hydrocarbon
chain radical having from 2 to 10 carbon atoms having one or two
olefinic double bonds, preferably one olefinic double bond. The
term "C.sub.2-N alkenyl" refers to an alkenyl comprising 2 to N
carbon atoms where N is an integer having the following values: 3,
4, 5, 6, 7, 8, 9, or 10. The term "C.sub.2-10 alkenyl" refers to an
alkenyl comprising 2 to 10 carbon atoms. Examples include, but are
not limited to vinyl, 1-propenyl, 2-propenyl, (allyl) or 2-butenyl
(crotyl).
[0097] The term "halogenated alkenyl" refers to an alkenyl
comprising at least one of F, Cl, Br, and I.
[0098] The term "alkyl" refers to an unbranched or branched chain,
saturated, monovalent hydrocarbon residue containing 1 to 30 carbon
atoms. The term "C.sub.1-N alkyl" refers to an alkyl comprising 1
to N carbon atoms, where N is an integer having the following
values: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30. The term
"C.sub.1-4" alkyl refers to an alkyl contain 1 to 4 carbon atoms.
The term "low alkyl" or "lower alkyl" denotes a straight or
branched chain hydrocarbon residue comprising 1 to 8 carbon atoms.
"C.sub.1-20 alkyl" as used herein refers to an alkyl comprising 1
to 20 carbon atoms. "C.sub.1-10 alkyl" as used herein refers to an
alkyl comprising 1 to 10 carbon atoms. Examples of alkyl groups
include, but are not limited to, methyl, ethyl, propyl, i-propyl,
n-butyl, i-butyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl,
heptyl, and octyl. The term (ar)alkyl or (heteroaryl)alkyl indicate
the alkyl group is optionally substituted by an aryl or a
heteroaryl group respectively.
[0099] The term "halogenated alkyl" (or "haloalkyl") refers to an
unbranched or branched chain alkyl comprising at least one of F,
Cl, Br, and I. The term "C.sub.1-3 haloalkyl" refers to a haloalkyl
comprising 1 to 3 carbons and at least one of F, Cl, Br, and I. The
term "halogenated lower alkyl" refers to a haloalkyl comprising 1
to 8 carbon atoms and at least one of F, Cl, Br, and I. Examples
include, but are not limited to, fluoromethyl, chloromethyl,
bromomethyl, iodomethyl, difluoromethyl, dichloromethyl,
dibromomethyl, diiodomethyl, trifluoromethyl, trichloromethyl,
tribromomethyl, triiodomethyl, 1-fluoroethyl, 1-chloroethyl,
1-bromoethyl, 1-iodoethyl, 2-fluoroethyl, 2-chloroethyl,
2-bromoethyl, 2-iodoethyl, 2,2-difluoroethyl, 2,2-dichloroethyl,
2,2-dibromoethyl, 2,2-diiodoethyl, 3-fluoropropyl, 3-chloropropyl,
3-bromopropyl, 3-iodopropyl, 2,2,2-trifluoroethyl,
1,1,2,2,2-pentafluoroethyl, 1-fluoro-1-chloroethyl, or
1-fluoro-1-chloro-1-bromoethyl.
[0100] The term "alkynyl" refers to an unbranched or branched
hydrocarbon chain radical having from 2 to 10 carbon atoms,
preferably 2 to 5 carbon atoms, and having one triple bond. The
term "C.sub.2-N alkynyl" refers to an alkynyl comprising 2 to N
carbon atoms, where N is an integer having the following values: 2,
3, 4, 5, 6, 7, 8, 9, or 10. The term "C.sub.2-4 alkynyl" refers to
an alkynyl comprising 2 to 4 carbon atoms. The term "C.sub.2-10
alkynyl" refers to an alkynyl comprising 2 to 10 carbon atoms.
Examples include, but are not limited to, ethynyl, 1-propynyl,
2-propynyl, 1-butynyl, 2-butynyl, or 3-butynyl.
[0101] The term "halogenated alkynyl" refers to an unbranched or
branched hydrocarbon chain radical having from 2 to 10 carbon atoms
preferably 2 to 5 carbon atoms, and having one triple bond and at
least one of F, Cl, Br, and I.
[0102] The term "cycloalkyl" refers to a saturated carbocyclic ring
comprising 3 to 8 carbon atoms, i.e. cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl. The term
"C.sub.3-7 cycloalkyl" as used herein refers to a cycloalkyl
comprising 3 to 7 carbons in the carbocyclic ring.
[0103] The term "alkoxy" refers to an --O-alkyl group, wherein
alkyl is defined above. Examples include, but are not limited to,
methoxy, ethoxy, n-propyloxy, i-propyloxy, n-butyloxy, i-butyloxy,
t-butyloxy. "Lower alkoxy" or "low alkoxy" or "low alkoxyl" as used
herein denotes an alkoxy group with a "lower alkyl" group as
previously defined. "C.sub.1-10 alkoxy" refers to an --O-alkyl
wherein alkyl is C.sub.1-10.
[0104] The term "halogenated alkoxy" refers to an --O-alkyl group
in which the alkyl group comprises at least one of F, Cl, Br, and
I.
[0105] The term "halogenated lower alkoxy" or "halogenated low
alkoxy" refers to an --O-(lower alkyl) group in which the lower
alkyl group comprises at least one of F, Cl, Br, and I.
[0106] The term "substituted", as used herein, means that one or
more hydrogens on the designated atom is replaced with a selection
from the indicated group, provided that the designated atom's
normal valency is not exceeded, and that the substitution results
in a stable compound.
[0107] The term "protected", as used herein and unless otherwise
defined, refers to a group that is added to an oxygen, nitrogen, or
phosphorus atom to prevent its further reaction or for other
purposes. A wide variety of oxygen and nitrogen protecting groups
are known to those skilled in the art of organic synthesis.
Non-limiting examples include: C(O)-alkyl, C(O)Ph, C(O)aryl,
CH.sub.3, CH.sub.2-alkyl, CH.sub.2-alkenyl, CH.sub.2Ph,
CH.sub.2-aryl, CH.sub.2O-alkyl, CH.sub.2O-aryl, SO.sub.2-alkyl,
SO.sub.2-aryl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl,
and 1,3-(1,1,3,3-tetraisopropyldisiloxanylidene).
[0108] The term "halo" as used herein includes fluoro, chloro,
bromo, and iodo.
[0109] The term "purine" or "pyrimidine" base includes, but is not
limited to, adenine, N-alkylpurines, N.sup.6-acylpurines (wherein
acyl is C(O)(alkyl, aryl, alkylaryl, or arylalkyl),
N.sup.6-benzylpurine, N.sup.6-halopurine, N.sup.6-vinyulpurine,
N.sup.6-acetylenic purine, N.sup.6-acyl purine,
N6-hydroxyalkylpurine, N.sup.6-allylaminopurine, N.sup.6-thioallyl
purine, N.sup.2-alkylpurines, N.sup.2-alkyl-6-thiopurines, thymine,
cytosine, 5-fluorocytosine, 5-methylcytosine, 6-azapyrimidine,
including 6-azacytosine, 2- and 4-mercaptopyrimidine, uracil,
5-halouracil, including 5-fluorouracil, C.sup.5-alkylpyrimidines,
C.sup.5-benzylpyrimidines, C.sup.5-halopyrimidines,
C.sup.5-vinylpyrimidine, C.sup.5-acetylenic pyrimidine,
C.sup.5-acyl pyrimidine, C.sup.5-hydroxyalkyl purine,
C.sup.5-aminopyrimidine, C.sup.5-cyanopyrimidine,
C.sup.5-iodopyrimidine, C.sup.6-iodo-pyrimidine, C.sup.5--Br-vinyl
pyrimidine, C.sup.6--Br-vinyl pyrimidine, C.sup.5-nitropyrimidine,
C.sup.5-amino-pyrimidine, N.sup.2-alkylpurines,
N.sup.2-alkyl-6-thiopurines, 5-azacytidinyl, 5-azauracilyl,
triazolopyridinyl, imidazolopyridinyl, pyrrolopyrimidinyl, and
pyrazolopyrimidinyl. Purine bases include, but are not limited to,
guanine, adenine, hypoxanthine, 2,6-diaminopurine, and
6-chloropurine. Functional oxygen and nitrogen groups on the base
can be protected as necessary or desired. Suitable protecting
groups are well known to those skilled in the art, and include
trimethylsilyl, dimethylhexylsilyl, t-butyldimethylsilyl, and
t-butyldiphenylsilyl, trityl, alkyl groups, and acyl groups such as
acetyl and propionyl, methansulfonyl, and p-toluenesulfonyl.
[0110] The term "tautomerism" and "tautomers" have their accepted
meanings.
[0111] The term "pharmaceutically acceptable salt or prodrug" is
used throughout the specification to describe any pharmaceutically
acceptable form (such as an ester, phosphate ester, salt of an
ester or related group) of a compound which upon administration to
a mammal, provides the active compound. Pharmaceutically acceptable
salts include those derived from pharmaceutically acceptable
inorganic or organic bases and acids. Pharmaceutically acceptable
prodrugs refer to a compound that is metabolized, for example
hydrolyzed or oxidized, in the host to form a compound of the
present invention. Typical examples of prodrugs include compounds
that have biologically labile protecting groups on a functional
moiety of the selected compound. Prodrugs include compounds that
can be oxidized, reduced, aminated, deaminated, hydroxylated,
dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated,
acylated, deacylated, phosphorylated, dephosphorylated to produce
the active compound. The compounds of this invention possess
antiviral activity against the HIV, HBV and HCV viruses, or are
metabolized to a compound that exhibits such activity.
[0112] In cases where compounds are sufficiently basic or acidic to
form stable nontoxic acid or base salts, administration of the
compound as a pharmaceutically acceptable salt may be appropriate.
Examples of pharmaceutically acceptable salts are organic acid
addition salts formed with acids, which form a physiological
acceptable anion, for example, tosylate, methanesulfonate, acetate,
citrate, malonate, tartarate, succinate, benzoate, ascorbate,
.alpha.-ketoglutarate, and .alpha.-glycerophosphate. Suitable
inorganic salts may also be formed, including sulfate, nitrate,
bicarbonate and carbonate salts.
[0113] Alternatively, pharmaceutically acceptable salts may be
obtained, for example, by reacting a sufficiently basic compound
such as an amine with a suitable acid affording a physiologically
acceptable anion. Alkali metal (for example, sodium, potassium, or
lithium) or alkaline earth metal (for example, calcium and
magnesium) salts of, for example, carboxylic acids can also be
made.
[0114] Any of the compounds described herein can be administered as
a prodrug to increase the activity, bioavailability, stability or
otherwise alter the properties of the selected compound. A number
of prodrug ligands are known.
[0115] The term "host" as used herein, refers to a unicellular or
multicellular organism in which the virus can replicate, including
but not limited to cell lines and animals, and preferably a human.
Alternatively, the host can be carrying a part of the viral genome,
whose replication or function can be altered by the compounds of
the present invention. The term host specifically refers to
infected cells, cells transfected with all or part of the viral
genome and animals.
[0116] The compounds of the present invention may be formulated in
a wide variety of oral administration dosage forms and carriers.
Oral administration can be in the form of tablets, coated tablets,
hard and soft gelatin capsules, solutions, emulsions, syrups, or
suspensions. Compounds of the present invention are efficacious
when administered by suppository administration, among other routes
of administration. The most convenient manner of administration is
generally oral using a convenient daily dosing regimen which can be
adjusted according to the severity of the disease and the patient's
response to the antiviral medication.
[0117] A compound or compounds of the present invention, as well as
their pharmaceutically acceptable salts, together with one or more
conventional excipients, carriers, or diluents, may be placed into
the form of pharmaceutical compositions and unit dosages. The
pharmaceutical compositions and unit dosage forms may be comprised
of conventional ingredients in conventional proportions, with or
without additional active compounds and the unit dosage forms may
contain any suitable effective amount of the active ingredient
commensurate with the intended daily dosage range to be employed.
The pharmaceutical compositions may be employed as solids, such as
tablets or filled capsules, semisolids, powders, sustained release
formulations or liquids such as suspensions, emulsions, or filled
capsules for oral use; or in the form of suppositories for rectal
or vaginal administration. A typical preparation will contain from
about 5% to about 95% active compound or compounds (w/w). The term
"preparation or "dosage form" is intended to include both solid and
liquid formulations of the active compound and one skilled in the
art will appreciate that an active ingredient can exist in
different preparations depending on the desired dose and
pharmacokinetic parameters.
[0118] The term "excipient" as used herein refers to a compound
that is used to prepare a pharmaceutical composition, and is
generally safe, non-toxic and neither biologically nor otherwise
undesirable, and includes excipients that are acceptable for
veterinary use as well as human pharmaceutical use. The compounds
of this invention can be administered alone but will generally be
administered in admixture with one or more suitable pharmaceutical
excipients, diluents or carriers selected with regard to the
intended route of administration and standard pharmaceutical
practice.
[0119] A "pharmaceutically acceptable salt" form of an active
ingredient may also initially confer a desirable pharmacokinetic
property on the active ingredient which was absent in the non-salt
form, and may even positively affect the pharmacodynamics of the
active ingredient with respect to its therapeutic activity in the
body. The phrase "pharmaceutically acceptable salt" of a compound
as used herein means a salt that is pharmaceutically acceptable and
that possesses the desired pharmacological activity of the parent
compound. Such salts include: (1) acid addition salts, formed with
inorganic acids such as hydrochloric acid, hydrobromic acid,
sulfuric acid, nitric acid, phosphoric acid, and the like; or
formed with organic acids such as glycolic acid, pyruvic acid,
lactic acid, malonic acid, maleic acid, fumaric acid, tartaric
acid, citric acid, 3-(4-hydroxybenzoyl)benzoic acid,
1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid,
benzenesulfonic acid, 4-chlorobenzenesulfonic acid,
2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic
acid, lauryl sulfuric acid, gluconic acid, glutamic acid,
salicyclic acid, muconic acid, and the like or (2) basic addition
salts formed with the conjugate bases of any of the inorganic acids
listed above, wherein the conjugate bases comprise a cationic
component selected from among Na.sup.+, K.sup.+, Mg.sup.+2,
Ca.sup.+2, NHgR'''4-g+, in which R''' is a C.sub.1-3 alkyl and g is
a number selected from among 0, 1, 2, 3, or 4. It should be
understood that all references to pharmaceutically acceptable salts
include solvent addition forms (solvates), water addition forms
(hydrates), or crystal forms (polymorphs) as defined herein, of the
same acid additions salts.
[0120] Solid form preparations include powders, tablets, pills
capsules, suppositories, and dispersible granules. A solid carrier
may be one or more substances which may also act as diluents,
flavoring agents, solubilizers, lubricants, suspending agents,
binders, preservatives, tablet disintegrating agents, or an
encapsulating material. In powders, the carrier generally is a
finely divided solid which is a mixture with the finely divided
active component. In tablets, the active component generally is
mixed with the carrier having the necessary binding capacity in
suitable proportions and compacted in the shape and size desired.
Suitable carriers include but are not limited to magnesium
carbonate, magnesium stearate, talc, sugar, lactose, pectin,
dextrin, starch, gelatin, tragacanth, methylcellulose, sodium
carboxymethylcellulose, a low melting wax, cocoa butter, and the
like. Solid form preparations may contain, in addition to the
active component colorants, flavors, stabilizers, buffers,
artificial and natural sweeteners, dispersants, thickeners,
solubilizing agents, and the like.
[0121] Liquid formulations also are suitable for oral
administration include liquid formulations including emulsions,
syrups, elixirs and aqueous suspensions. These include solid form
preparations which are intended to be converted to liquid form
preparations shortly before use. Emulsions may be prepared in
solutions, for example, in aqueous propylene glycol solutions or
may contain emulsifying agents such as lecithin, sorbitan
monooleate, or acacia. Aqueous suspensions can be prepared by
dispersing the finely divided active component in water with
viscous material, such as natural or synthetic gums, resins,
methylcellulose, sodium carboxymethylcellulose, and other well
known suspending agents.
[0122] The compounds of the present invention may be formulated for
administration as suppositories. A low melting wax, such as a
mixture of fatty acid glycerides or cocoa butter is first melted
and the active component is dispersed homogeneously, for example,
by stirring. The molten homogeneous mixture is then poured into
convenient sized molds, allowed to cool and to solidify.
[0123] The compounds of the present invention may be formulated for
vaginal administration. Pessaries, tampons, creams, gels, pastes,
foams or sprays containing in addition to the active ingredient
such carriers as are known in the art to be appropriate.
[0124] Suitable formulations along with pharmaceutical carriers,
diluents and excipients are described in Remington: The Science and
Practice of Pharmacy 1995, edited by E. W. Martin, Mack Publishing
Company, 19th Edition, Easton, Pa., which is hereby incorporated by
reference. A skilled formulation scientist may modify the
formulations within the teachings of the specification to provide
numerous formulations for a particular route of administration
without rendering the compositions of the present invention
unstable or comprising their therapeutic activity.
[0125] The modification of the present compounds to render them
more soluble in water or other vehicle, for example, may be easily
accomplished by minor modifications (e.g., salt formulation), which
are well within the ordinary skill in the art. It is also well
within the ordinary skill of the art to modify the route of
administration and dosage regimen of a particular compound in order
to manage the pharmacokinetics of the present compounds for maximum
beneficial effect in patients.
[0126] The term "medicament" means a substance used in a method of
treatment and/or prophylaxis of a subject in need thereof, wherein
the substance includes, but is not limited to, a composition, a
formulation, a dosage from, and the like, comprising a compound of
formula I. It is contemplated that the use of the compound
represented by formula I in the manufacture of a medicament for the
treatment of any of the antiviral conditions disclosed herein can
be any of the compounds contemplated in any of the aspects of the
invention, either alone or in combination with other compounds of
the present invention.
[0127] The term "subject" means a mammal, which includes, but is
not limited to, cattle, pigs, sheep, chicken, turkey, buffalo,
llama, ostrich, dogs, cats, and humans, preferably the subject is a
human.
[0128] The term "therapeutically effective amount" as used herein
means an amount required to reduce symptoms of the disease in an
individual. The dose will be adjusted to the individual
requirements in each particular case. That dosage can vary within
wide limits depending upon numerous factors such as the severity of
the disease to be treated, the age and general health condition of
the patient, other medicaments with which the patient is being
treated, the route and form of administration and the preferences
and experience of the medical practitioner involved. For oral
administration, a daily dosage of between about 0.1 and about 10 g,
including all values in between, such as 0.25, 0.5, 0.75, 1, 1.5,
2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, and 9.5
g, per day should be appropriate in monotherapy and/or in
combination therapy. A preferred daily dosage is between about 0.5
and about 7.5 g per day, a more preferred dosage is between 1.5 and
about 6.0 g per day. Generally, treatment is initiated with a large
initial "loading dose" to rapidly reduce or eliminate the virus
followed by a decreasing of the dose to a level sufficient to
prevent resurgence of the infection. One of ordinary skill in
treating diseases described herein will be able, without undue
experimentation and in reliance on personal knowledge, experience,
and the disclosures of this application, to ascertain a
therapeutically effective amount of the compounds of the present
invention for a given disease and patient.
[0129] Therapeutic efficacy in HBV and HCV treatment can be
ascertained from tests of liver function including, but not limited
to protein levels such as serum proteins (e.g., albumin, clotting
factors, alkaline phosphatase, aminotransferases (e.g., alanine
transaminase, aspartate transaminase), 5'-nucleosidase,
C-glutaminyltranspeptidase, etc.), synthesis of bilirubin,
synthesis of cholesterol, and synthesis of bile acids; a liver
metabolic function, including, but not limited to, carbohydrate
metabolism, amino acid and ammonia metabolism. Alternatively the
therapeutic effectiveness may be monitored by measuring HBV or
HCV-RNA the results of these tests will allow the dose to be
optimized. For HIV, therapeutic efficacy in HIV infection can be
ascertained by measuring HIV-RNA levels from plasma samples and
measuring levels of CD4 cells.
[0130] The disclosed compounds or their pharmaceutically acceptable
derivatives or salts or pharmaceutically acceptable formulations
containing these compounds are useful in the prevention and
treatment of HIV infections and other related conditions such as
AIDS-related complex (ARC), persistent generalized lymphadenopathy
(PGL), AIDS-related neurological conditions, anti-HIV antibody
positive and HIV-positive conditions, Kaposi's sarcoma,
thrombocytopenia purpurea and opportunistic infections. In
addition, these compounds or formulations can be used
prophylactically to prevent or retard the progression of clinical
illness in individuals who are anti-HIV antibody or HIV-antigen
positive or who have been exposed to HIV.
[0131] Another aspect of the present invention comprises
administering a therapeutically effective amount of a compound
represented by formula I and a therapeutically effective amount of
another antiviral agent; wherein the administration is concurrent
or alternative or sequential. It is understood that the time
between alternative (or sequential) administration can range
between 1-24 hours, which includes any sub-range in between
including 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, 20, 21, 22, and 23 hours.
[0132] It is contemplated that the another antiviral agent includes
but is not limited to interferon-.alpha., interferon-B, pegylated
interferon-.alpha., ribavirin, levovirin, viramidine, another
nucleoside HIV, HBV, or HCV polymerase inhibitor, an HIV, HBV, or
HCV non-nucleoside polymerase inhibitor, an HIV, HBV, or HCV
protease inhibitor, an HIV, HBV, or HCV helicase inhibitor or an
HIV, HBV, or HCV fusion inhibitor. When an active compound or its
derivative or salt are administered in combination with another
antiviral agent the activity may be increased over the parent
compound. When the treatment is combination therapy, such
administration may be concurrent or sequential with respect to that
of the nucleoside derivatives. "Concurrent administration" as used
herein thus includes administration of the agents at the same time
or at different times. Administration of two or more agents at the
same time can be achieved by a single formulation containing two or
more active ingredients or by substantially simultaneous
administration of two or more dosage forms with a single active
agent.
[0133] In another embodiment for the treatment of HIV infection,
the active compound or its prodrug or pharmaceutically acceptable
salt can be administered in combination or alternation with another
antiviral agent, such as another active anti-HIV agent, including
but not limited to those of the formulae above, others listed below
or known in the art. In general, in combination therapy, effective
dosages of two or more agents are administered together, whereas
during alternation therapy, an effective dosage of each agent is
administered serially. The dosage will depend on absorption,
inactivation, and excretion rates of the drug as well as other
factors known to those of skill in the art. It is to be noted that
dosage values will also vary with the severity of the condition to
be alleviated. It is to be further understood that for any
particular subject, specific dosage regimens and schedules should
be adjusted over time according to the individual need and the
professional judgment of the person administering or supervising
the administration of the compositions.
[0134] Nonlimiting examples of antiviral agents that can be used in
combination with the compounds disclosed herein include the
following: Invirase.RTM., Fortovase.RTM., Norvir.RTM.,
Crixivan.RTM., Viracept.RTM., Agenerase.RTM., Kaletra.RTM.,
Retrovir.RTM., Epivir.RTM., Combivir.RTM., Triazivir.RTM.,
Ziagen.RTM., Hivid.RTM., Videx.RTM., Didex.RTM. EC, Zerit.RTM.,
Viread.RTM., Covincil.TM., Viramune.RTM., Rescriptor.RTM.,
Sustiva.RTM., Droxia.RTM., Fuzeon.RTM., Atazanavir.RTM.,
Proleukin.RTM., Remune.RTM., Procrit.RTM., Darunavir.RTM., and
Serostim.RTM.,
Experimental Results
[0135] It will be understood that references herein to treatment
extend to prophylaxis as well as to the treatment of existing
conditions. Furthermore, the term "treatment" of a viral infection,
as used herein, also includes treatment or prophylaxis of a disease
or a condition associated with or mediated by the viral infection,
or the clinical symptoms thereof.
[0136] Another embodiment is directed to a process for preparing
the compound represented by A or A', to the compound A or A'
obtained by the process, and to a composition comprising A or A'
obtained by the process, the process comprising
[0137] (1) deoxygenating the 2'-C-position of a compound
represented by 1 or 1'
##STR00035##
to obtain a compound represented by 2 or 2'
##STR00036##
and
[0138] (2) derivatizing the compound represented by 2 or 2' to
obtain a compound represented by A or A'
##STR00037## [0139] wherein for structures 1, 1', 2, 2', A, and
A'
[0140] (a) R.sup.2 is independently CH.sub.3, CH.sub.2F, CHF.sub.2,
CF.sub.3, F, CN, C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, or C.sub.1-4
alkyl optionally substituted with amino, hydroxy, or 1 to 3
fluorine atoms;
[0141] (b) R is H, phosphate, including 5'-monophosphate,
5',3'-cyclic phosphate, diphosphate, triphosphate, or a stabilized
phosphate prodrug, H-phosphonate, including stabilized
H-phosphonates, acyl, including optionally substituted phenyl and
lower acyl, alkyl, including lower alkyl, O-substituted
carboxyalkylamino or its peptide derivatives, sulfonate ester,
including alkyl or arylalkyl sulfonyl, including methanesulfonyl
and benzyl, wherein the phenyl group is optionally substituted, a
lipid, including a phospholipid, an L or D-amino acid, a
carbohydrate, a peptide, a cholesterol, or other pharmaceutically
acceptable leaving group which when administered in vivo;
[0142] (c) R.sup.3 is independently OH, H, C.sub.1-4 alkyl,
C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, vinyl, N.sub.3, CN, Cl, Br,
F, I, NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4 alkyl),
C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl), O(C.sub.1-10
acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH, S(C.sub.1-4
acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl), S(C.sub.2-4
alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl), SO(C.sub.2-4
alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4 acyl),
SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.1-4 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2,
N(C.sub.1-18 acyl).sub.2, wherein alkyl, alkynyl, alkenyl and vinyl
are optionally substituted by N.sub.3, CN, one to three halogen
(Cl, Br, F, I), NO.sub.2, C(O)O(C.sub.1-4 alkyl), C(O)O(C.sub.1-4
alkyl), C(O)O(C.sub.2-4 alkynyl), C(O)O(C.sub.2-4 alkenyl),
O(C.sub.1-4 acyl), O(C.sub.1-4 alkyl), O(C.sub.2-4 alkenyl), SH,
S(C.sub.1-4 acyl), S(C.sub.1-4 alkyl), S(C.sub.2-4 alkynyl),
S(C.sub.2-4 alkenyl), SO(C.sub.1-4 acyl), SO(C.sub.1-4 alkyl),
SO(C.sub.2-4 alkynyl), SO(C.sub.2-4 alkenyl), SO.sub.2(C.sub.1-4
acyl), SO.sub.2(C.sub.1-4 alkyl), SO.sub.2(C.sub.2-4 alkynyl),
SO.sub.2(C.sub.2-4 alkenyl), OS(O).sub.2(C.sub.1-4 acyl),
OS(O).sub.2(C.sub.14 alkyl), OS(O).sub.2(C.sub.2-4 alkenyl),
NH.sub.2, NH(C.sub.1-4 alkyl), NH(C.sub.2-4 alkenyl), NH(C.sub.2-4
alkynyl), NH(C.sub.1-4 acyl), N(C.sub.1-4 alkyl).sub.2, N(C.sub.1-4
acyl).sub.2;
[0143] (d) R.sup.4 is independently H, a lower alkyl, CN, vinyl,
O-(lower alkyl), hydroxyl lower alkyl, i.e., --(CH.sub.2).sub.pOH,
where p is 1-6, including hydroxylmethyl (CH.sub.2OH), CH.sub.2F,
N.sub.3, CH.sub.2CN, CH.sub.2NH.sub.2, CH.sub.2NHCH.sub.3,
CH.sub.2N(CH.sub.3).sub.2, ethynyl alkyne (optionally substituted),
or halogen, including F, Cl, Br, or I, alkenyl, alkynyl, Br-vinyl,
hydroxy, O-alkenyl, NO.sub.2, amino, loweralkylamino, or
di(loweralkyl)amino;
[0144] (e) R and R.sup.3 can together form 5',3'-cyclic phosphate
including stabilized prodrugs thereof;
[0145] (f) Base is a naturally occurring or modified purine or
pyrimidine base represented by the following structures:
##STR00038##
[0146] wherein for a and b
[0147] Z is Nor CR.sup.8;
[0148] R.sup.5, R.sup.6, and R.sup.7 are independently H, F, Cl,
Br, I, OH, OR', SH, SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form
saturated or unsaturated rings, or saturated or unsaturated
heterocyclic rings), lower alkyl of C.sub.1-C.sub.6, halogenated
(F, Cl, Br, I) lower alkyl of C.sub.1-C.sub.6, lower alkenyl of
C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower alkenyl of
C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6 such as
C.ident.CH, halogenated (F, Cl, Br, I) lower alkynyl of
C.sub.2-C.sub.6, lower alkoxy of C.sub.1-C.sub.6, halogenated (F,
Cl, Br, I) lower alkoxy of C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R',
CONH.sub.2, CONHR', CONR.sub.12, CH.dbd.CHCO.sub.2H, or
CH.dbd.CHCO.sub.2R' wherein R' is an optionally substituted alkyl,
which includes, but is not limited to, an optionally substituted
C.sub.1-20 alkyl, an optionally substituted C.sub.1-10 alkyl, an
optionally substituted lower alkyl; an optionally substituted
cycloalkyl; an optionally substituted alkynyl of C.sub.2-C.sub.6,
an optionally substituted lower alkenyl of C.sub.2-C.sub.6, or
optionally substituted acyl, which includes but is not limited to
C(O) alkyl, C(O)(C.sub.1-20 alkyl), C(O)(C.sub.1-10 alkyl), or
C(O)(lower alkyl), optionally substituted aryl, optionally
substituted C.sub.1-C.sub.4 alkylaryloxy, heteroaryl, optionally
substituted C.sub.1-C.sub.4 alkyl-heteroaryl, an optionally
substituted alkoxy C.sub.1-20 alkyl, an optionally substituted
amino C.sub.1-20 alkyl, an optionally substituted fluoro C.sub.1-20
alkyl;
[0149] R.sup.8 is independently H, halogen (including F, Cl, Br,
I), OH, OR', SH, SR', NH.sub.2, NHR', NR'.sub.2 (two R' can form
saturated or unsaturated rings, or saturated or unsaturated
heterocyclic rings), NO.sub.2, lower alkyl of C.sub.1-C.sub.6,
halogenated (F, Cl, Br, I) lower alkyl of C.sub.1-C.sub.6, lower
alkenyl of C.sub.2-C.sub.6, halogenated (F, Cl, Br, I) lower
alkenyl of C.sub.2-C.sub.6, lower alkynyl of C.sub.2-C.sub.6,
halogenated (F, Cl, Br, I) lower alkynyl of C.sub.2-C.sub.6, lower
alkoxy of C.sub.1-C.sub.6, halogenated (F, Cl, Br, I) lower alkoxy
of C.sub.1-C.sub.6, CO.sub.2H, CO.sub.2R', CONH.sub.2, CONHR',
CONR.sub.12, CH.dbd.CHCO.sub.2H, or CH.dbd.CHCO.sub.2R', wherein R'
is an optionally substituted alkyl, which includes, but is not
limited to, an optionally substituted C.sub.1-20 alkyl, an
optionally substituted C.sub.1-10 alkyl, an optionally substituted
lower alkyl; an optionally substituted cycloalkyl; an optionally
substituted alkynyl of C.sub.2-C.sub.6, an optionally substituted
lower alkenyl of C.sub.2-C.sub.6, or optionally substituted acyl,
which includes but is not limited to C(O) alkyl, C(O)(C.sub.1-20
alkyl), C(O)(C.sub.1-10 alkyl), or C(O)(lower alkyl), optionally
substituted aryl, optionally substituted C.sub.1-C.sub.4
alkyl-aryloxy, heteroaryl, optionally substituted C.sub.1-C.sub.4
alkyl-heteroaryl, an optionally substituted alkoxy C.sub.1-20
alkyl, an optionally substituted amino C.sub.1-20 alkyl, an
optionally substituted fluoro C.sub.1-20 alkyl; or
base may be selected from a group of formula c
##STR00039##
wherein for structure c, 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;
[0150] 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, CNNR, 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.sup.X 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 linesindicate a possible pi or double bond;
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; or base may be a structure
selected from the group consisting of structures d-n
##STR00040## ##STR00041##
wherein Z, X, and R'' are defined as in structure c; base may be a
structure selected from the group consisting of structures o-ff
##STR00042## ##STR00043## ##STR00044##
wherein G and R are defined as in structure c; base may be a
structure gg
##STR00045##
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
defined as in structure c; base may be a structure hh
##STR00046##
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 in
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 defined
as in structure c; base may be a structure ii
##STR00047##
wherein R and G are defined as in structure c; base may be a
structure jj
##STR00048##
wherein R and G are defined as in structure c; or base may be a
structure kk
##STR00049##
wherein for structure kk: [0151] R is selected from the group
consisting of hydrogen and C.sub.1-C.sub.3 alkyl; [0152] X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
[0153] Y is selected from the group consisting of a bond, O, and
CH.sub.2; [0154] Q is absent or is selected from the group
consisting of O, S, and NH, provided that when Q is absent, V and
NH are both attached to a CH.sub.2 group; [0155] V is selected from
the group consisting of N and C-G; [0156] Z is selected from the
group consisting of N and C-G'; [0157] G and G' are independently
selected from the group consisting of hydrogen, amino,
aminocarbonyl, methylamino, dimethylamino, acylamino, alkoxyamino,
--SO.sub.3H, --SO.sub.2NH.sub.2, aminocarbonylamino,
oxycarbonylamino, HR'NCHR''C(O)NH--, azido, cyano, halo,
hydroxyamino, and hydrazino, where R' is hydrogen and R'' is a
side-chain of an amino acid or where R' and R'' together with the
nitrogen and carbon bound to each group respectively form a
pyrrolidinyl group; [0158] provided that V and Z are not identical;
[0159] provided that when V is C--H, Z is N; [0160] T.sup.1 and
T.sup.2 are independently selected from the group consisting of
hydrogen, hydroxyl, C.sub.1-C.sub.4-alkoxy,
C.sub.1-C.sub.4-thioalkoxy, amino, substituted amino, and halo; and
[0161] each of W, W.sup.1, and W.sup.2 is independently selected
from the group consisting of hydrogen, C.sub.1-C.sub.4 alkyl, and a
prodrug group; or base may be a structure ll
##STR00050##
[0161] wherein for structure ll: R is C.sub.1-C.sub.3 alkyl; X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
Q' is selected from the group consisting of NH, O, and S; G' is
selected from the group consisting of amino, aminocarbonyl,
methylamino, dimethylamino, acylamino, --SO.sub.3H,
--SO.sub.2NH.sub.2, alkoxyamino, aminocarbonylamino,
oxycarbonylamino, HR'NCHR''C(O)NH--, azido, cyano, halo,
hydroxyamino, and hydrazino, where R' is hydrogen and R'' is a
side-chain of an amino acid or where R' and R'' together with the
nitrogen and carbon bound to each group respectively form a
pyrrolidinyl group; Y is selected from the group consisting of a
bond, O, and CH.sub.2; and each of W, W.sup.1, and W.sup.2 is
independently selected from the group consisting of hydrogen,
C.sub.1-C.sub.4 alkyl, and a prodrug group; or base may be a
structure mm
##STR00051##
where in for structure mm [0162] Q is as defined for structure kk
[0163] A and B are independently selected from the group consisting
of C=Q, NH, and methylene optionally substituted with 1 to 2 halo
groups, provided that A and B are not both NH; [0164] D is NH, or
-D-A-B- together form a --N.dbd.CH--NH--, --(C=Q)-CH.sub.2--(C=Q)-,
--(C=Q)-NH--(C=Q)-, --(CX')=(CX')--(C=Q)-, or --CH.dbd.CH--NH--
group where X' is halo; [0165] each Q is independently selected
from the group consisting of O, S, and NH; R is selected from the
group consisting of hydrogen and C.sub.1-C.sub.3 alkyl; [0166] X is
selected from the group consisting of hydrogen, halo, and OW.sup.2;
[0167] T.sup.1 and T.sup.2 are independently selected from the
group consisting of hydrogen, hydroxyl, C.sub.1-C.sub.4-alkoxy,
C.sub.1-C.sub.4-thioalkoxy, amino, substituted amino, and halo;
[0168] Y is selected from the group consisting of a bond, O, and
CH.sub.2; and each of W, W.sup.1, and W.sup.2 is independently
selected from the group consisting of hydrogen, C.sub.1-C.sub.4
alkyl, and a prodrug group; and pharmaceutically acceptable salts,
tautomers, pharmaceutically acceptable salts of tautomers, salts
(acidic or basic addition salts), hydrates, solvates, crystalline
forms thereof, optionally in combination with one or more
antiviral, antibacterial, or antiproliferative agents.
[0169] Not to be limited by way of example, the following
exemplified embodiments are intended to convey information related
to methods of making and using the disclosed and claimed
compounds.
Compound Preparation
General Preparation of 2'-(R)-2'-C-Methyl-2'-deoxynucleosides
[0170] The preparation of 2'-substituted-2'-deoxynucleosides is
illustrated in Scheme 1 below. Treatment of compound 1 with TIPDSCl
followed by reaction of the resulting intermediate with mono-methyl
oxalyl chloride gives compound 3. Treatment of 3 with
AIBN/n-BuSn.sub.3H followed by desilylation with TBAF provides
2'-substituted-2'-deoxynucleoside 6. Intermediate 4 also can be
prepared by treatment of 2 with PhOCSCl followed by reductive
deoxygenation as used for 4 from 3.B and R.sup.2' are defined as
above.
##STR00052##
Preparation of 2'-(R)-2'-C-Methyl-2'-deoxyuridine
[0171] Above general method for the preparation of
2'-substituted-2'-deoxynucleosides is exemplified in Scheme 2 for
the synthesis of 2-(R)-2'-C-methyl-2'-deoxyuridine. Treatment of
2'-C-methyl-uridine with TIPDSCl in pyridine or in the presence of
imidazole followed by reaction of the resulting intermediate 8 with
mono-methyl oxalyl chloride gave compound 9 in good yield.
Deoxygenation of 9 was accomplished by treatment of 9 with
AIBN/n-Bu.sub.3SnH to give 10, which upon deprotection gave
2'-(R)-2'-C-methyl-2'-deoxyuridine (11).
##STR00053##
General Preparation of
2'-(R)-4'-Azido-2'-C-methyl-2'-deoxynucleosides
[0172] General synthesis of
2'-(R)-4'-azido-2'-C-methyl-2'-deoxynucleosides is shown in Scheme
3. Treatment of compound 12 with iodine in the presence of
triphenylphosphine followed by elimination in the presence of base,
such as, NaOMe or DBU or the like, gives 4'-methylene-nucleoside
14. Protection of 3'-OH of 14 by treatment with TBSCl in the
presence of imidazole provides compound 15. Epoxidation of 15
followed by ring-opening by treatment of epoxide with TMSN.sub.3 in
the presence of SnCl.sub.4 also affords 4'-azido-nucleoside 18
after deprotection of 17 by TBAF. Compound 18 also can be prepared
by azizo-iodination of 14. Treatment of 14 with ICl and NaN.sub.3
gives 4'-azido-5'-iodo-nucleoside 19 in good yield. 3'-OH
protection with BzCl followed by 5'-iodo-oxidation with
m-chloroperbenzoic acid in the presence of m-chlorobenzoic acid
provides the protected nucleoside 20. Deprotection of 20 also
affords 4'-azido-nucleosides 18.
##STR00054##
Preparation of 2'-(R)-4'-Azido-2'-C-methyl-2'-deoxycytidine (Scheme
4)
[0173] Scheme 4 illustrated the preparation of
2'-(R)-4'-azido-2'-C-methyl-2'-deoxycytidine. Treatment of compound
11 with iodine in the presence of triphenylphosphine followed by
elimination catalyzed by NaOMe in MeOH gave compound 22 in good
yield. Reaction of 22 with IN.sub.3 provided intermediate 27.
Protection of 3'-OH of 27 by treatment with BzCl in pyridine
followed by oxidation of 5'-iodine with mCPBA in the presence of
mCBA afforded protected uridine analog 28. The target nucleoside 26
was prepared by treatment of 28 with triisopropylbenzenesulfonyl
chloride in the presence of DMAP followed by ammonium hydroxide
then methanolic ammonia. Compound 26 can also be prepared through
expoxide intermediate 24 by treatment of compound 23 with
DMDO/acetone followed by ring-opening with TMSN.sub.3 in the
presence of SnCl.sub.4 and conversion of uridine analog to cytidine
derivative.
##STR00055##
Preparation of Aldehyde Intermediates 49 and 50 (Scheme 5)
[0174] Treatment of compound 42 with DMTrCl in pyridine in the
presence of DMAP followed by TBSCl/imidazole selectively gave
compound 44 in excellent yield. Deoxygentation of compound 44
##STR00056##
was accomplished by treatment with methyl chlorooxoacetate in the
presence of DMAP followed by AIBN/(TMS).sub.3SiH in toluene to
provide 2'-deoxynucleoside intermediate 45. Detritylation of 45
followed by Dess-Martin oxidation afforded aldehyde 47. Compound 47
was subject to Aldol-condensation followed by reduction with
NaBH.sub.4 gave diol 48. Selective oxidation of 48 by Dess-Martin
reagent provided intermediate 49. 5'-Hydroxy of 49 was protected
with silyl to afforded intermediate 50.
Preparation of 4'-C-cyano-2'-methyl-2'-deoxycytidine (Scheme 6)
[0175] Treatment of compound 49 with hydroxylamine hydrochloride in
pyridine followed by heating with acetic anhydride at 120.degree.
C. in the presence of sodium acetate gave compound 52. Amination of
compound 52 was accomplished by treatment of 52 with tosyl chloride
in the presence of N-methylpiperidine and triethylamine followed by
ammonium hydroxide to afford cytidine intermediate 53. Desilylation
of 53 with TEAF and acetylation followed by treatment with
methanole ammonia produced compound 55.
##STR00057##
Preparation of 4'-C-vinyl-2'-methyl-2'-deoxycytidine (Scheme 7)
[0176] Treatment of aldehyde 50 with methyltriphosphonium chloride
in the presence of LiBu gave 4'-C-vinyl intermediate 56. Compound
58 was prepared by similar fashion. Further elimination of 58 to
4'-C-ethynyl analog was accomplished by treatment of 58 with LiBu.
Using similar method for preparation of compound 53 from 52,
amination of compounds 56 and 59 gave compounds 57 and 60,
respectively.
##STR00058##
Preparation of 4'-C-hydroxymethyl-2'-methyl-2'-deoxycytidine
(Scheme 8)
[0177] Acetylation of compound 48 with acetyl anhydride gave fully
protected intermediate 61. Similarly, amination of 61 followed by
deprotection provided 4'-C-hydroxymethyl-2'-methyl-2'-deoxycytidine
62.
##STR00059##
Preparation of 4'-C-allyl- and 4'-C-cyano-2'-methylthymidine
(Scheme 9)
[0178] Epoxidation of compound 63 with DMDO followed by treatment
of the resulting epoxide by allyltrimethylsilane in the presence of
SnCl.sub.4 gave silyl protected intermediate 65. Treatment of 65
with methanolic ammonia followed by ammonium fluoride in MeOH
provided 4'-C-allyl-nucleoside 69.
[0179] Similarly, compound 70 was prepared from intermediate 64
when trimethylsilyl cyanide was used as nucleophile for the epoxide
opening.
##STR00060##
Preparation of 4'-C-ethynyl-2'-methylthymidine (Scheme 10)
[0180] Treatment of compound 64 with tri(allyl)alumina followed by
desilylation gave products 73 and 74 after separation.
##STR00061##
Experiment Details:
Synthesis of Compound 28
##STR00062##
[0182] Compound 27 was prepared from starting nucleoside,
2'-C-methyl-2'-deoxyuridine, by treatment with I.sub.2/Ph.sub.3P
and elimination catalyzed by NaOMe followed by azido-iodination
with NCl/NaN.sub.3.
[0183] To a solution of the alcohol (203.7 mg 0.53 mmol, 1.0 eq.)
in dichloromethane (DCM) was added triethylamine (TEA) (148 .mu.l,
1.06 mmol, 2.0 eq.) and dimethylaminopyridine (DMAP, catalytic
amount). After 5 minutes, benzoyl chloride (BzCl, 68 .mu.l, -0.58
mmol, 1.1 eq.) was added and the reaction was monitored by LCMS
(liquid chromatography-mass spectroscopy). After 10 minutes the
reaction was complete. Water and NaHCO.sub.3 were added and the
mixture was extracted with DCM (2.times.), the organic layer was
washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated. Purification over silica-gel eluted with
EtOAC-Heptane from 3:7 to 6:4 afforded protected iodide (180 mg,
68%) of product as a white solid.
##STR00063##
[0184] To a solution of iodide (180 mg, 0.362 mmol, 1.0 eq.) in DCM
(18 ml) and water (9 ml) were added successively K.sub.2HPO.sub.4
(126 mg, 0.724 mmol, 2.0 eq.), nBu.sub.4NHSO.sub.4 (135 mg, 0.398
mmol, 1.1 eq.) and mCBA (meta-Chlorobenzoic acid) (72 mg, 0.398
mmol, 1.1 eq.). The reaction mixture was cooled to 0.degree. C. and
mCPBA (meta-Chloroperbenzoic acid) 77% (243 mg, 1.086 mmol, 3.0
eq.) was added and the reaction was left to reach room temperature.
After 14 hours LCMS indicated completion of the reaction. Saturated
aqueous NaHCO.sub.3 and Na.sub.2SO.sub.3 were added to the reaction
mixture and the mixture was extracted with EtOAc (100 ml). The
organic layer was washed with brine and dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by FCC over silica-gel, eluted with EtOAc-Heptane from 7:3
to 5:5 to yield 145 mg (76%) of product 28 as a white solid.
Synthesis of Compound 26
##STR00064##
[0186] A mixture of POCl.sub.3 (94 .mu.l, 1.009 mmol, 4.0 eq.) and
triazole (331 mg, 4.793 mmol, 19.0 eq.) in dry MeCN (5 ml) was
stirred at 0.degree. C. for 5 minutes followed by a slow addition
of TEA (0.74 ml). The resulting mixture was left at 0.degree. C.
for 1 hour and then a solution of the uridine (28, 163 mg, 0.252
mmol, 1.0 eq.) in dry MeCN (5 ml) was added. The reaction mixture
was stirred at room temperature for 14 hours then filtered through
a pad of celite and the solid was washed with 3 ml of MeCN. EtOAc
was added (70 ml) and the solution was washed with saturated
aqueous NaHCO.sub.3, water, brine, and then the solution was
concentrated. The residue was co-evaporated with dioxane (20 ml).
The residue was used in the next step without purification.
##STR00065##
[0187] To a solution of the triazole (159 mg) in MeOH (5 ml) was
added sodium methoxide (.about.240 .mu.l, 25% wt. in MeOH, 4.0
eq.). After 30 minutes, LCMS showed the reaction was complete. To
the reaction mixture was added HCl (1.1 ml, 1N, 4.0 eq.) and the
mixture was concentrated. The residue was purified over silica gel,
eluted with DCM-MeOH from 1% to 6% to yield 44 mg (54%) of the
product as an oil.
##STR00066##
[0188] The methoxy compound (40 mg, 135 mmol, 1.0 eq.) was
dissolved in 0.5N ammonia in dioxane (5 ml). The reaction mixture
was heated to 120.degree. C. in a microwave (250 W, 150 PSI) for 3
hours. Progress of the reaction was followed by LCMS. Upon
completion of the reaction the solvent was removed and the residue
was purified over silica gel eluted with DCM-EtOH from 95:5 to
80:20 to yield 16.8 mg of the desired product 26 as a syrup.
[0189] The products listed below are prepared using a similar
procedure described above.
TABLE-US-00003 Product H-NMR data of products MS Starting material
##STR00067## (360 MHz, CD3OD): 1.33 (d, J = 25.2 Hz, 3H), 3.80 (m,
2H), 4.13 (d, J = 21.6 Hz, 1H), 5.94 (d, J = 7.2 Hz, 1H), 6.53 (d,
J = 10.8 Hz, 1H), 8.08 (d, J = 6.1 Hz, 1H). M + 1 = 301.13 (calc
300.10) for C10H13FN6O4) ##STR00068## ##STR00069## (360 MHz,
CD3OD): 3.80 (m, 2H), 4.57 (dd, J = 3.6, 21.6 Hz, 1H), 5 (dd, J =
3.6, 50.4 Hz, 1H), 5.94 (d, J = 7.2 Hz, 1H), 6.16 (d, J = 21.6 Hz,
1H), 7.93 (d, J = 7.2 Hz, 1H). M + 1 = 287.10 (calc 286.08) for
C9H11FN6O4) ##STR00070## ##STR00071## (360 MHz, CD3OD): 1.39 (d, J
= 22.3 Hz, 1H), 3.80 (m, 1H), 4.20 (d, 25.2 Hz, 1H), 5.74 (d, J =
7.2 Hz, 1H), 6.40 (d, J = 14.4 Hz, 1H), 7.95 (d, J = 8.2 Hz, 1H). M
+ 1 = 302.12 (calc 301.08) for C10H12FN5O5) ##STR00072##
##STR00073## (360 MHz, CD3OD): 3.76 (m, 2H), 4.59 (dd, J = 3.6,
21.6 Hz, 1H), 5.22 (dd, J = 3.6, 54.0 Hz, 1H), 5.70 (d, J = 10.8
Hz, 1H), 6.16 (dd, J = 1.4, 18.0 Hz, 1H), 7.85 (d, J = 8.2 Hz, 1H).
M + 1 = 288.10 (calc 287.07) for C9H10FN5O5) ##STR00074##
##STR00075## (400 MHz, CD3OD): 3.90 (dd J = 12.4 Hz, 2H), 4.50 (t,
12.8 Hz, 1H), 5.93 (d, J = 7.6 Hz, 1H), 6.40 (br. S, 1H), 7.71 (d,
J = 7.6 Hz, 1H). M + 1 = 305.10 (calc 304.07) for C9H10F2N6O4)
##STR00076## ##STR00077## (400 MHz, DMSO-d6): 4.52 (m, 1H), 5.75
(d, J = 8.0 Hz, 1H), 5.86 (t, J = 6.0 Hz, 1H), 6.20 (t, J = 7.6 Jz,
1H), 6.78 (d, 6.8 Hz, 1H), 7.68 (d, J = 8.0 Hz, 1H), 11.66 (s, 1H).
M + 1 = 306.05 (calc 305.06) for C9H9F2N5O5) ##STR00078##
##STR00079## (250 MHz, CD3OD): 0.93 (d, J = 2.5 Hz, 3H), 2.70 (m,
1H), 3.89 (m, 3H), 5.89 (d, J = 7.5 Hz, 1H), 6.44 (m, 1H), 7.95 (d,
J = 7.5 Hz, 1H). M + 1 = 283.12 (calc 282.11) for C10H14N6O4)
##STR00080## ##STR00081## (250 MHz, CD3OD): 0.99 (d, J = 5.0 Hz,
3H), 2.73 (m, 1H), 3.90 (m, 2H), 4.04 (d, J = 12.5 Hz, 1H), 5.72
(d, J = 7.5 Hz, 1H), 6.37 (d, J = 10.0 Hz, 1H), 7.95 (d, J = 10.0
Hz, 1H). M + 1 = 284.29 (calc 283.09) for C10H13N5O5) ##STR00082##
##STR00083## (250 MHz, CD3OD): 3.85 (s, 2H), 4.50 (dd, 5.0, 22.5
Hz, 1H), 5.20 (td, 5.0, 55.0 Hz, 1H), 5.73 (d, J = 7.5 Hz, 1H),
6.46 (dd, J = 7.5, 12.5 Hz, 1H), 7.75 (dd, 2.5, 7.5 Hz, 1H). M + 1
= 288.19 (calc 287.07) for (C9H10FN5O5) ##STR00084## ##STR00085##
(250 MHz, CD3OD): 3.85 (s, 2H), 4.49 (dd, 5.0, 22.5 Hz, 1H), 5.22
(td, 2.5, 57.5 Hz, 1H), 5.93 (d, J = 7.5 Hz, 1H), 6.49 (dd, J =
5.0, 12.5 Hz, 1H), 7.76 (d, 7.5 Hz, 1H). M + 1 = 287.12 (calc
286.08) for C9H11FN6O4) ##STR00086## ##STR00087## (250 MHz, CD3OD):
0.96 (d, J = 7.5 Hz, 3H), 2.70 (m, 1H), 3.84 (dd, J = 12.5 Hz, 1H),
4.03 (d, J = 10.0 Hz, 1H), 6.37 (d, J = 7.5 Hz, 1H), 8.25 (7.5 Hz,
1H). M + 1 = 301.16 (calc 300.10) for C10H13FN6O4) ##STR00088##
##STR00089## (250 MHz, CD3OD): 1.02 (d, J = 7.5 Hz, 3H), 2.71 (m,
1H), 3.92 (dd, 12.5 Hz, 1H), 4.07 (d, J = 10.0 Hz, 1H), 6.34 (d, J
= 7.5 Hz, 1H), 8.30 (J = 7.5 Hz, 1H). M + Na = 323.90 (calc 324.08
for C10H12FN5O5Na) ##STR00090## ##STR00091## (450 MHz, CDC13): 0.97
(d, J = 7.2 Hz, 3H), 1.86 (d, J = 1.2 Hz, 3H), 2.08 (d, OH), 2.19
(t, OH), 2.68 (m, 1H), 3.84 (d, J = 12 Hz, 1H), 3.92 (d, J = 12 Hz,
1H), 4.07 (d, J = 11.2 Hz, 1H), 6.33 (d, J = 7.6 Hz, 1H), 7.83 (s,
1H). M + 1 = 298.10 (calcd 297.11 for C11H15N5O5 ##STR00092##
Synthesis of Compound 43
##STR00093##
[0191] To a solution of compound 42 (1.1 g, 4.26 mmol) in anhydrous
pyridine (50 mL) was added DMTrCl (2.17 g, 6.39 mmol) at rt. The
mixture was stirred for 5 h, diluted with EtOAc (100 mL), washed
with water (25 ml.times.4), and dried over sodium sulfate. After
filtration and concentration, the residue was co-evaporated with
toluene (30 mL) and purified by flash column chromatography (MeOH
in CH.sub.2Cl.sub.2 0 to 5%) to give 43 (1.9 g, 80%). .sup.1H NMR
(CDCl.sub.3) .delta. (ppm) 9.94 (s, 1H, NH), 8.17 (d, 1H, J=8.0 Hz,
H-6), 7.40-7.24 (m, 9H, aromatic), 6.84 (m, 4H, aromatic), 6.08 (s,
1H, H-1'), 5.28 (dd, 1H, J=2.0, 8.4 Hz, H-5), 4.80 (s, 1H, HO),
4.11-4.01 (m, 2H, H-3' and 4'), 3.79 (s, 6H, (OCH.sub.3).times.2),
3.61 (dd, 1H, J=2.4, 11.6 Hz, H-5'), 3.55 (dd, 1H, J=2.4, 11.2 Hz,
H-5''), 2.87 (d, 1H, J=9.2 Hz, HO), 1.32 (s, 3H, CH.sub.3).
Synthesis of Compound 44
##STR00094##
[0193] To a solution of 43 (1.9 g, 3.39 mmol) and imidazole (0.69
g, 10.17 mmol) in anhydrous CH.sub.2Cl.sub.2 (20 mL) was added
TBSCl (0.77 g, 5.08 mmol) at rt. The resulting mixture was stirred
at rt for 48 h. Additional imidazole (0.69 g) and TBSCl (0.77 g)
were added. Then the mixture was stirred at rt for 72 h, diluted
with CH.sub.2Cl.sub.2 (80 mL), washed with water (40 mL.times.2),
and dried over sodium sulfate. After filtration and concentration,
the residue was purified by flash silica gel column chromatography
(0-20-35% EtOAc in hexanes) to give compound 44 (2 g, 87%) as a
white solid. .sup.1H NMR (CDCl.sub.3) .delta. (ppm) 8.34 (s, 1H,
NH), 8.16 (d, 1H, J=8.4 Hz, H-6), 7.32-7.17 (m, 9H, aromatic), 6.85
(m, 4H, aromatic), 6.13 (s, 1H, H-1'), 5.08 (dd, 1H, J=2.4, 8.0 Hz,
H-5), 4.19 (d, 1H, J=8.4 Hz, H-3'), 4.01 (m, 1H, H-4'), 3.87 (dd,
1H, J=2.0, 10.8 Hz, H-5'), 3.80 (s, 6H, (OCH.sub.3).times.2), 3.32
(dd, 1H, J=2.0, 10.8 Hz, H-5''), 1.19 (s, 3H, CH.sub.3), 0.79 (s,
9H, C(CH.sub.3).sub.3), 0.07 (s, 3H, CH.sub.3Si), -0.30 (s, 3H,
CH.sub.3Si).
Synthesis of Compound 45
##STR00095##
[0195] To a solution of 44 (2.0 g, 2.96 mmol), DAMP (2.17 g, 17.78
mmol), and triethylamine (2.48 mL, 17.78 mmol) in acetonitrile (40
mL) was added methyl chlorooxoacetate (1.64 mL, 17.78 mmol)
dropwise under ice-water bath. The resulting mixture was stirred at
rt for 1 h, diluted with EtOAc (125 mL), washed with brine (30
mL.times.4), and dried over sodium sulfate. After filtration and
concentration, the residue was co-evaporated with toluene (20
mL.times.2) and dried under high vacuum for 10 min. Then the
residue was dissolved in anhydrous toluene (40 mL) and bubbled with
nitrogen gas for 10 min, to which TMSsilylhydride (5.49 mL, 17.78
mmol) and then AIBN (1.46 g, 8.89 mmol) were added. The resulting
mixture was refluxed in preheated oil bath at 120.degree. C. for
1.5 h and concentrated under reduced pressure. The residue was
purified by flash column chromatography (0-20-35% EtOAc in hexanes)
to give compound 45 (1.3 g, 67%) as a white solid. .sup.1H NMR
(CDCl.sub.3) .delta. (ppm) 8.15 (d, 1H, J=8.4 Hz, H-6), 8.02 (s,
1H, NH), 7.34-7.20 (m, 9H, aromatic), 6.84 (m, 4H, aromatic), 6.30
(d, 1H, J=7.6 Hz, H-1'), 5.11 (dd, 1H, J=2.4, 8.4 Hz, H-5), 4.14
(d, 1H, J=8.0 Hz, H-3'), 3.83-3.74 (m, 8H, H-4', 5' and
(CH.sub.3O).sub.2), 3.33 (dd, 1H, J=2.4, 10.8 Hz, H-5''), 2.54 (m,
1H, H-2'), 0.98 (d, 3H, J=6.8 Hz, CH.sub.3), 0.76 (d, 9H,
C(CH.sub.3).sub.3), 0.04 (s, 3H, CH.sub.3Si), -0.28 (s, 3H,
CH.sub.3Si).
Synthesis of Compound 46
##STR00096##
[0197] To a solution of compound 45 (1.3 g, 1.97 mmol) in anhydrous
CH.sub.2Cl.sub.2 (10 mL) was added TFA (0.3 mL, 3.95 mmol) under
ice-water bath. The resulting mixture was stirred at rt for 1 h,
diluted with CH.sub.2Cl.sub.2 (100 mL), washed with sat.NaHCO.sub.3
(30 mL.times.2), and dried over sodium sulfate. After filtration
and concentration, the residue was purified by silica gel column
chromatography (EtOAc:Hexanes=1:1) to give compound 46 (600 mg,
85%) as a white solid.
Synthesis of Compound 47
##STR00097##
[0199] To a solution of compound 46 (3 g, 8.46 mmol) in anhydrous
CH.sub.2Cl.sub.2 (30 mL) was added 15% Dess-Martin periodinane in
CH.sub.2Cl.sub.2 (36 mL, 12.73 mmol) dropwise at 0.degree. C. The
reaction mixture was stirred at 0.degree. C. for 3 h, diluted with
CH.sub.2Cl.sub.2 (150 mL), washed with sodium bicarbonate solution
(50 mL.times.3) and then with saturated solution of sodium
thiosulfate (50 mL.times.3), and the organic layer dried over
sodium sulfate. After filtration and concentration, the residue was
purified by silica gel column chromatography
(MeOH:CH.sub.2Cl.sub.2=1:20) to give compound 47. .sup.1H NMR
(CDCl.sub.3) .delta. (ppm) 9.79 (s, 1H, H-5'), 8.86 (s, 1H, NH),
8.26 (d, 1H, J=7.2 Hz, H-6), 6.15 (d, 1H, J=6.0 Hz, H-1'), 5.77 (d,
1H, J=8.0 Hz, H-5), 4.40 (d, 1H, J=3.6 Hz, H-4'), 4.26 (t, 1H,
J=3.6 Hz, H-3'), 2.70 (m, 1H, H-2'), 0.92 (s, 9H,
C(CH.sub.3).sub.3), 0.76 (d, 3H, J=7.6 Hz, CH.sub.3), 0.14 (s, 3H,
CH.sub.3Si), 0.13 (s, 3H, CH.sub.3Si).
Synthesis of Compound 48
##STR00098##
[0201] The compound 47 obtained was then dissolved in dioxane (100
mL) and 37% formaldehyde (3 mL, 36.96 mmol) was added. To the
obtained solution was added 2N sodium hydroxide (5 mL, 10 mmol)
dropwise at rt. The resulting reaction mixture was stirred at rt
for 15 h and cooled. Then sodium borohydride (890 mg, 23.53 mmol)
was added portionwise. The resulting mixture was stirred at rt for
6 h and AcOH-pyridine solution (2.5:7.5 mL) and water (100 mL) were
added under ice-water. The mixture was then extracted with
CH.sub.2Cl.sub.2 (50 mL.times.4) and the organic layer dried over
sodium sulfate. After filtration and concentration, the residue was
purified silica gel column chromatography
(MeOH:CH.sub.2Cl.sub.2=1:40 to 1:20) to give compound 48 (2.1 g,
64%) as a white solid. .sup.1H NMR (CD.sub.3OD) .delta. (ppm) 8.30
(d, 1H, J=8.0 Hz, H-6), 6.31 (d, 1H, J=8.0 Hz, H-1'), 5.68 (d, 1H,
J=8.0 Hz, H-5), 4.29 (d, 1H, J=9.6 Hz, H-3'), 3.86 (d, 1H, J=12.0
Hz, CH-4'), 3.78 (d, 1H, J=12.0 Hz, CH-4'), 3.54 (d, 1H, J=12.0 Hz,
CH-4'), 3.45 (d, 1H, J=12.0 Hz, CH-4'), 2.83 (m, 1H, H-2'),
0.98-0.88 (m, 12H, CH.sub.3-2' and C(CH.sub.3).sub.3), 0.15 (s, 3H,
CH.sub.3Si), 0.13 (s, 3H, CH.sub.3Si).
Synthesis of Compound 49
##STR00099##
[0203] To a solution of compound 48 (1.4 g, 3.62 mmol) in
CH.sub.2Cl.sub.2-THF (46:10 mL) was added Dess-Martin periodinane
(1.8 g, 4.24 mmol) at 0.degree. C. in one portion. The mixture was
stirred at 0.degree. C. for 2 h. After temperature was raised to
10.degree. C., the mixture was stirred for 1 h and sodium
thiosulfate (1.0 g) was added. Then the mixture was stirred for 30
min and added onto top of a silica gel column, which was then
eluted with CH.sub.2Cl.sub.2 and CH.sub.2Cl.sub.2-EtOAc (3:1) to
give compound 49 (a mixture of desired .alpha./.beta.=2) as a white
solid (900 mg, 64%).
Synthesis of Compound 50
##STR00100##
[0205] To a solution of compound 49 (500 mg, 1.30 mmol) and
imidazole (530 mg, 7.8 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added
TBSCl (590 mg, 3.90 mmol) at rt. The resulting mixture was stirred
for 3 h, diluted with CH.sub.2Cl.sub.2, washed with water, and
dried over sodium sulfate. After filtration and concentration, the
residue was separated by flash column chromatography (EtOAc 0 to
25% in hexanes) to give compound 50 (270 mg, 40%).
Synthesis of Compound 51
##STR00101##
[0207] To a solution of compound 48 (220 mg, 0.57 mmol) in
anhydrous CH.sub.2Cl.sub.2-THF (10 mL:2 mL) was added Dess-Martin
periodinane (300 mg, 0.71 mmol) in one portion at 0.degree. C. The
resulting reaction mixture was stirred for 2 h and sodium
thiosulfate (390 mg) was added. The mixture was stirred for 15 min
at 0.degree. C., then poured onto top of a short silica gel column,
and eluted with CH.sub.2Cl.sub.2-EtOAc (1:1) thoroughly. The
fractions were combined and concentrated in vacuo to a residue,
which was then dissolved in pyridine (10 mL) and NH.sub.2OH--HCl
(300 mg) added. The resulting mixture was stirred at rt for 15 h
and concentrated in vacuo. The residue obtained was purified by
silica gel column chromatography (MeOH:CH.sub.2Cl.sub.2=1:40 to
1:20) to give compound 51 (107 mg, 47%) along with 4'-.beta.-isomer
(53 mg, 18%) as white solids. .sup.1H NMR (CD.sub.3OD) .delta.
(ppm) 8.20 (d, 1H, J=8.0 Hz, H-6), 7.4 (s, 1H, HC.dbd.N), 6.31 (d,
1H, J=8.0 Hz, H-1'), 5.70 (d, 1H, J=8.0 Hz, H-5), 4.32 (d, 1H,
J=10.0 Hz, H-3'), 3.84 (s, 2H, H-5'), 2.64 (m, 1H, H-2'), 0.94 (d,
3H, J=7.2 Hz, CH.sub.3), 0.92 (s, 9H, C(CH.sub.3).sub.3), 0.17 (s,
3H, CH.sub.3Si), 0.13 (s, 3H, CH.sub.3Si); .sup.1H NMR (CD.sub.3OD)
for minor isomer .delta. (ppm) 7.71 (d, 1H, J=8.0 Hz, H-6), 7.55
(s, 1H, HC.dbd.N), 6.29 (d, 1H, J=7.6 Hz, H-1'), 5.70 (d, 1H, J=8.0
Hz, H-5), 4.37 (d, 1H, J=8.0 Hz, H-3'), 3.92 (d, 1H, J=12.4 Hz,
H-5'), 3.73 (d, 1H, J=12.4 Hz, H-5''), 2.86 (m, 1H, H-2'),
0.93-0.88 (m, 12H, CH.sub.3-2' and C(CH.sub.3).sub.3), 0.12 (s, 3H,
CH.sub.3Si), 0.10 (s, 3H, CH.sub.3Si).
Synthesis of Compound 52
##STR00102##
[0209] A mixture of compound 51 (160 mg, 0.40 mmol) and sodium
acetate (123 mg, 1.5 mmol) in acetic anhydride (5 mL) was heated at
120.degree. C. for 3 h and concentrated in vacuo. The obtained
residue was chromatographed with a silica gel column by eluting
with MeOH--CH.sub.2Cl.sub.2 (1:40) to give compound 52 (97 mg, 57%)
as a white solid. .sup.1H NMR (CDCl.sub.3) .delta. (ppm) 10.03 (s,
1H, NH), 7.26 (d, 1H, J=8.0 Hz, H-6), 6.17 (bs, 1H, H-1'), 5.78 (d,
1H, J=8.0 Hz, H-5), 4.52 (d, 1H, J=12.4 Hz, H-5'), 4.42 (d, 1H,
J=12.4 Hz, H-5''). 4.19 (bs, 1H, H-3'), 2.86 (m, 1H, H-2'), 2.17
(s, 3H, CH.sub.3CO), 1.03 (d, 1H, J=7.2, CH.sub.3), 0.95 (s, 9H,
C(CH.sub.3).sub.3), 0.15 (s, 6H, CH.sub.3Si).
Synthesis of Compound 53
##STR00103##
[0211] To a solution of compound 52 (200 mg, 0.47 mmol) in
anhydrous CH.sub.3CN (4 mL) containing triethylamine (0.20 mL, 1.42
mmol) and N-methylpiperidine (0.11 mL, 0.94 mmol) was added TsCl
(270 mg, 1.42 mmol) at rt. The resulting mixture was stirred at rt
for 1 h and then 29% NH.sub.4OH (4 mL) was added under water bath.
The resulting mixture was stirred at rt for 2 h and concentrated
under 35.degree. C. The obtained residue was purified by silica gel
column chromatography (MeOH:CH.sub.2Cl.sub.2=1:10 to 1:4) to give
compound 53 (160 mg, 89%) as a syrup.
Synthesis of Compound 54
##STR00104##
[0213] A solution of compound 53 (160 mg, 0.42 mmol) and TEAF (200
mg, 1.34 mmol) in MeOH-THF (2:4 mL) was stirred at rt for 15 h and
heated at 60.degree. C. for 4 h. After concentration, the residue
was purified by silica gel column chromatography
(MeOH:CH.sub.2Cl.sub.2=1:10 to 1:4) to give crude compound 55 (100
mg, crude, contaminated by TEAF), which was then dissolved in
anhydrous pyridine, then treated with acetic anhydride, stirred at
rt for 3 h, and concentrated in vacuo. The obtained residue was
purified by silica gel column chromatography
(MeOH:CH.sub.2Cl.sub.2=1:40) to give compound 54 as a syrup which
was used in next reaction. .sup.1H NMR (CD.sub.3OD) .delta. (ppm)
7.97 (d, 1H, J=7.6 Hz, H-6), 7.46 (d, 1H, J=7.6 Hz, H-5), 6.23 (bs,
1H, H-1'), 5.52 (bs, 1H, H-3'), 4.65 (d, 1H, J=12.0 Hz, H-5'), 4.62
(d, 1H, J=12.0 Hz, H-5''), 3.11 (m, 1H, H-2'), 2.19 (s, 3H,
CH.sub.3CO), 2.19 (s, 3H, CH.sub.3CO), 2.12 (s, 3H, CH.sub.3CO),
0.97 (d, 3H, J=7.2 Hz, CH.sub.3); MSES (M+1):393.
Synthesis of Compound 55
##STR00105##
[0215] Compound 54 was dissolved in 7M ammonia in methanol (5 mL),
stirred in a sealed flask for 15 h, and concentrated in vacuo. The
obtained residue was chromatographed with silica gel column by
eluting with MeOH--CH.sub.2Cl.sub.2 (1:4) to give compound 55 (30
mg, 27% from 53) as a white solid. UV(.lamda..sub.max) 273 nm
(MeOH); .sup.1H NMR (CD.sub.3OD) .delta. (ppm) 7.85 (d, 1H, J=7.8
Hz, H-6), 6.40 (bs, 1H, H-1'), 5.88 (d, 1H, J=7.6 Hz, H-5), 4.10
(d, 1H, J=9.6 Hz, H-3'), 4.00 (d, 1H, J=12.4 Hz, H-5'), 3.92 (d,
1H, J=12.4 Hz, H-5''), 2.75 (m, 1H, H-2'), 0.95 (d, 3H, J=3.2 Hz,
CH.sub.3); MSES (M+1): 267.
Synthesis of Compound 56
##STR00106##
[0217] To a suspension of methyltriphenylphosphonium bromide in THF
(2 mL) was added n-BuLi (2.2 M in hexanes, 0.24 mL, 0.528 mmol) in
anhydrous THF (2 mL) at -78.degree. C. The mixture was stirred at
0.degree. C. for 1 h and compound 50 (60 mg, 0.12 mmol) in
anhydrous THF (2 mL) was added. The mixture was then stirred at rt
for 1 h, neutralized with saturated aqueous ammonium chloride
solution, diluted with EtOAc, washed with brine, and dried over
sodium sulfate. After filtration and concentration, the residue was
purified by silica gel column chromatography
(MeOH:CH.sub.2Cl.sub.2=1:40) to give compound 56 as a white solid
(45 mg, 75%).
Synthesis of Compound 57
##STR00107##
[0219] To a solution of compound 56 (100 mg, 0.20 mmol) in
anhydrous CH.sub.3CN (2 mL) containing triethylamine (0.08 mL, 0.6
mmol) and N-methylpiperidine (0.05 mL, 0.40 mmol) was added TsCl
(120 mg, 0.6 mmol) at rt. The resulting mixture was stirred at rt
for 1 h and then 29% NH.sub.4OH (2 mL) was added under water bath.
The resulting mixture was stirred at rt for 2 h and concentrated in
vacuo under 35.degree. C. The obtained residue was purified by
silica gel column chromatography (MeOH:CH.sub.2Cl.sub.2=1:10 to
1:4) to give a protected cytidine intermediate (70 mg, 70%) as a
syrup. A mixture of the intermediate (70 mg, 0.14 mmol) and
ammonium fluoride (100 mg, 2.82 mmol) in methanol (10 mL) was
refluxed for 6 h and another 15 h after adding additional ammonium
fluoride (100 mg, 2.82 mmol). After concentration in vacuo, the
residue was purified by silica gel column chromatography
(MeOH:CH.sub.2Cl.sub.2=1:10 to 1:4) to give compound 57 (28.6 mg,
77%). UV(.lamda..sub.max) 273 nm (MeOH); .sup.1H NMR (CD.sub.3OD)
.delta. (ppm) 8.28 (d, 1H, J=7.2 Hz, H-6), 6.28 (d, 1H, J=7.6 Hz,
H-1'), 5.99 (dd, 1H, J=11.2, 17.2 Hz, H--C.dbd.C), 5.91 (d, 1H,
J=7.6 Hz, H-5), 5.49 (dd, 1H, J=2.0, 17.2 Hz, H--C.dbd.C), 5.32
(dd, 1H, J=2.0, 10.8 Hz, H--C.dbd.C), 4.06 (d, 1H, J=10.8 Hz,
H-3'), 3.72 (d, 1H, J=12.0 Hz, H-5'), 3.55 (d, 1H, J=12.0 Hz,
H-5''), 2.45 (m, 1H, H-2'), 0.87 (d, 3H, J=6.8 Hz, CH.sub.3); MSES:
268 (M+1), 535 (2M+1).
Synthesis of Compound 58
##STR00108##
[0221] To a suspension of chloromethyltriphenylphosphonium chloride
(390 mg, 1.12 mmol) in THF (3 mL) was added n-BuLi (2.2 M in
hexanes, 0.51 mL, 1.12 mmol) at -78.degree. C. The mixture was
stirred at 0.degree. C. for 1 h and compound 50 (140 mg, 0.28 mmol)
in anhydrous THF (3 mL) was added. The resulting mixture was
stirred at rt for 3 h, neutralized with saturated ammonium
chloride, diluted with EtOAc, washed with brine, and dried over
sodium sulfate. After filtration and concentration, the residue was
purified by silica gel column chromatography
(MeOH:CH.sub.2Cl.sub.2=1:40) to give compound 58 as a white solid
(140 mg, 94%).
Synthesis of Compound 59
##STR00109##
[0223] To a solution of compound 58 (200 mg, 0.38 mmol) in
anhydrous THF (10 mL) was added n-BuLi (2.8 mL 1.6 M in hexanes,
4.52 mmol) dropwise at -78.degree. C. The mixture was then stirred
at -78.degree. C. for 2 h, neutralized with saturated ammonium
chloride solution (10 mL), diluted with EtOAc (50 mL), washed with
brine (15 mL.times.3), and dried over sodium sulfate. After
filtration and concentration, the residue was purified by silica
gel column chromatography (hexanes:EtOAc=4:1) to give compound 59
(180 mg, 97%) as a white solid.); .sup.1H NMR (CDCl.sub.3) .delta.
(ppm) 9.21 (bs, 1H, NH), 8.05 (d, 1H, J=8.4 Hz, H-6), 6.24 (d, 1H,
J=7.6 Hz, H-1'), 5.94 (dd, 1H, J=11.2, 17.6 Hz, H--C.dbd.C), 5.70
(dd, 1H, J=2.0, 8.0 Hz, H--C.dbd.C), 5.52 (dd, 1H, J=1.2, 17.2 Hz,
H--C.dbd.C), 5.32 (d, 1H, J=8.4 Hz, H-5), 4.16 (d, 1H, J=10.4 Hz,
H-3'), 3.64 (d, 1H, J=11.2 Hz, H-5'), 3.57 (d, 1H, J=11.6 Hz,
H-5''), 2.48 (m, 1H, H-2'), 0.95 (s, 9H, C(CH.sub.3).sub.3), 0.91
and 0.90 (m, 12H, C(CH.sub.3).sub.3 and CH.sub.3-2'), 0.12-0.10 (4
s, 12H, CH.sub.3Si).
Synthesis of Compound 60
##STR00110##
[0225] To a solution of compound 59 (100 mg, 0.20 mmol) in
anhydrous CH.sub.3CN (3 mL) containing triethylamine (0.08 mL, 0.6
mmol) and N-methylpiperidine (0.05 mL, 0.40 mmol) was added TsCl
(120 mg, 0.61 mmol) at rt. The resulting mixture was stirred at rt
for 1 h and then 29% NH.sub.4OH (2 mL) was added under water bath.
The resulting mixture was stirred at rt for 2 h and concentrated in
vacuo under 35.degree. C. The obtained residue was purified by
silica gel column chromatography (MeOH:CH.sub.2Cl.sub.2=1:20 to
1:10) to give a cytidine intermediate (70 mg, 70%) as a syrup. A
mixture of the intermediate (70 mg, 0.14 mmol) and ammonium
fluoride (260 mg, 7.09 mmol) was heated at 90.degree. C. in a
sealed flask for 15 h and concentrated in vacuo. The residue was
purified by silica gel column chromatography
(MeOH:CH.sub.2Cl.sub.2=1:4) to give compound 60 (31.2 mg, 85%) as a
white solid. UV(.lamda..sub.max) 273 nm (MeOH); .sup.1H NMR
(CD.sub.3OD) .delta. (ppm) 8.05 (d, 1H, J=7.6 Hz, H-6), 6.31 (d,
1H, J=8.0 Hz, H-1'), 5.89 (d, 1H, J=7.6 Hz, H-5), 3.95 (d, 1H,
J=10.8 Hz, H-3'), 3.88 (d, 1H, J=12.4 Hz, H-5'), 3.79 (d, 1H,
J=12.4 Hz, H-5''), 3.06 (s, 1H, H--C--C), 2.75 (m, 1H, H-2'), 0.915
(d, 3H, J=6.8 Hz, CH.sub.3).
Synthesis of Compound 61
##STR00111##
[0227] To a solution of compound 48 (220 mg, 0.57 mmol) in
anhydrous pyridine (4 mL) was added acetic anhydride (0.27 mL, 2.85
mmol) at rt. The resulting mixture was stirred at rt for 5 h and
concentrated in vacuo. The resulting mixture was purified by silica
gel column chromatography (MeOH:CH.sub.2Cl.sub.2=1:40) to give
compound 61 (220 mg, 82%) as a white solid. .sup.1H NMR
(CDCl.sub.3) .delta. (ppm) 9.80 (bs, 1H, NH), 7.62 (d, 1H, J=8.0
Hz, H-6), 6.26 (d, 1H, J=7.2 Hz, H-1'), 5.74 (d, 1H, J=8.4 Hz,
H-5), 4.53 (d, 1H, J=12.4 Hz, HC-4'), 4.35 (d, 1H, J=12.0 Hz,
H-5'), 4.24 (d, 1H, J=12.0 Hz, H-5''), 4.05 (d, 1H, J=7.2 Hz,
H-3'), 3.99 (d, 1H, J=12.0 Hz, HC-4'), 2.82 (m, 1H, H-2''), 2.15
(s, 3H, CH.sub.3CO), 2.12 (s, 3H, CH.sub.3CO), 0.96 (d, 3H, J=7.2
Hz, CH.sub.3), 0.90 (s, 9H, C(CH.sub.3).sub.3), 0.11 (s, 3H,
CH.sub.3Si), 0.08 (s, 3H, CH.sub.3Si); MSES (M+1):266
Synthesis of Compound 62
##STR00112##
[0229] To a solution of compound 61 (220 mg, 0.47 mmol) in
anhydrous acetonitrile (5 mL) containing triethylamine (0.19 mL,
1.40 mmol) and N-methylpiperidine (0.11 mL, 0.93 mmol) was added
TsCl (270 mg, 1.40 mmol) at rt. The resulting mixture was stirred
at rt for 1 h and 29% NH.sub.4OH aqueous solution was added. Then
mixture was stirred at rt for 15 h and concentrated in vacuo. The
residue was purified by silica gel column chromatography
(MeOH:CH.sub.2Cl.sub.2=1:40) to give a cytidine intermediate (120
mg, 67%) as a white solid. A mixture of the intermediate (60 mg,
0.156 mmol) and ammonium fluoride (100 mg) in methanol (5 mL) was
heated at 90.degree. C. for 15 h and concentrated in vacuo. The
residue obtained was purified by silica gel column chromatography
(MeOH:CH.sub.2Cl.sub.2=1:10 to 1:4) to give compound 62 (10 mg,
24%) as a white solid. UV(.lamda..sub.max) 273 nm (MeOH); .sup.1H
NMR (D.sub.2O) .delta. (ppm) 8.08 (d, 1H, J=7.6 Hz, H-6), 6.32 (d,
1H, J=8.4 Hz, H-1'), 6.09 (d, 1H, J=7.6 Hz, H-5), 4.09 (d, 1H,
J=10.4 Hz, H-3'), 3.83 (d, 1H, J=12.8 Hz, CH-4'), 3.77 (d, 1H,
J=12.4 Hz, CH-4'), 3.72 (d, 1H, J=12.4 Hz, CH-4'), 3.60 (d, 1H,
J=12.4 Hz, CH-4'), 2.87 (m, 1H, H-2'), 0.87 (d, 3H, J=6.8 Hz,
CH.sub.3).
Synthesis of Compound 64
##STR00113##
[0231] To a solution of substrate 63 (150 mg, 0.43 mmol) in
CH.sub.2Cl.sub.2 (9 mL) was added cold solution of DMDO (11.33 mL,
1.8 eq, 0.79 mmol, from 0.07 M in acetone) at -30.degree. C. and
stirred for 1 h under argon atmosphere. The solvents were
evaporated and the residue was dried in vacuo with vigorous
stirring at 0.degree. C., and dried for additional 5 min. A viscous
solid 64 was formed and used for next step with out purification
(90%).
Synthesis of Compound 67
##STR00114##
[0233] To the cold solution of epoxide 64 (150 mg, 0.41 mmol) in
CH.sub.2Cl.sub.2 (10 mL) at -30.degree. C. were added
TMS-CH.sub.2CH.dbd.CH.sub.2 (90 mg, 146 uL, 1.22 mmol) followed by
SnCl.sub.4 (1.22 mL of 1 M solution in CH.sub.2Cl.sub.2, 1.22 mmol)
at once under argon atmosphere. The reaction mixture was stirred at
-30.degree. C. for 6 h and at room temperature for 2 h. The
reaction was quenched with sat Na.sub.2HCO.sub.3 solution, the
reaction mixture was filtered through a celite pad. The filtrate
was partitioned between CH.sub.2Cl.sub.2/H.sub.2O. The organic
layer was separated, dried (Na.sub.2SO.sub.4) and concentrated to
dryness. The crude product was treated with NH.sub.3/MeOH (5 mL, 7
N) for 15 h at room temperature. The solvent was removed in vacuo.
The residue was purified by silica gel chromatography, eluting with
10-40% ethyl acetate/hexane to afford compound 67 as colorless foam
(85 mg, 76%). Mass calcd: 410.22, Found: 411.10 (M.sup.++H);
.sup.1H NMR (CDCl.sub.3) .delta. 8.52 (s, NH), 7.76 (s, H), 6.19
(s, H), 5.89 (m, H), 5.15 (d, H, J=4 Hz), 5.11 (s, H), 4.25 (d, H,
J=9.2 Hz), 3.86 (d, H, J=11.2 Hz), 3.53 (dd, H, J=11.6 Hz, 3.2 Hz),
2.28 (m, H), 2.39 (dd, H, J=15.2 Hz, 6.8 Hz), 2.12 (dd, H, J=14.4
Hz, 8.4 Hz), 1.90 (s, 3H), 0.91 (s, 9H) 0.12 (s, 3H), 0.11 (s,
3H).
Synthesis of Compound 69
##STR00115##
[0235] To a well dried mixture of substrate 67 (30 mg, 0.073 mmol),
ammonium fluoride (32 mg, 0.73 mmol) was added methanol (3 mL) and
refluxed at 85.degree. C. for 12 h. Excess of ammonium fluoride (16
mg, 0.36 mmol) and methanol (2 mL) were added and continued
refluxing for additional 24 h. The reaction mixture was filtered
through a celite pad and the solvent was removed under reduced
pressure. The crude product was purified by silica gel
chromatography, eluting with 1-8% MeOH/CH.sub.2Cl.sub.2 to afford
product 69 as colorless solid (19 mg, 88%). Mass calcd: 296.32,
Found: 297.20 (M.sup.++H); .sup.1H NMR (CDCl.sub.3) .delta. 8.22
(s, H), 6.18 (d, H, J=8.4 Hz), 5.98 (m, H), 5.09 (m, 2H), 4.13 (d,
H, J=10.8 Hz), 3.76 (d, H, J=12.4 Hz), 3.55 (d, H, J=12 Hz), 2.66
(m, H), 2.39 (dd, H, J=14.8 Hz, 6.4 Hz), 2.17 (dd, H, J=14.6 Hz,
6.4 Hz), 1.85 (s, 3H), 0.93 (d, 3H, J=6.8 Hz).
Synthesis of Compound 68
##STR00116##
[0237] To the cold solution of epoxide 64 (120 mg, 0.33 mmol) in
CH.sub.2Cl.sub.2 (6 mL) at -30.degree. C. was added TMS-CN (60 mg,
0.65 mmol) followed by SnCl.sub.4 (0.65 mL, 0.65 mmol from 1 M
solution in CH.sub.2Cl.sub.2) at once under argon atmosphere. The
reaction mixture was stirred at -30.degree. C. for 1 h and stirred
at room temperature for 15 h. The reaction mixture was quenched
with sat Na.sub.2HCO.sub.3 solution. The reaction mixture was
filtered and the filtrate was partitioned between
CH.sub.2Cl.sub.2/H.sub.2O. The organic layer was separated, dried
(Na.sub.2SO.sub.4) and concentrated to dryness. The crude product
was treated with NH.sub.3/MeOH (7N, 5 mL) for 15 h at room
temperature. The solvent was removed and the crude product was
purified by silica gel chromatography, eluting with 4-30% ethyl
acetate/hexane to afford product 68 as colorless solid (72 mg,
56%). Mass calcd: 395.20, Found: 396.30 (M.sup.++H); .sup.1H NMR
(CDCl.sub.3) .delta. 8.63 (s, NH), 7.28 (s, H), 6.14 (d, H, J=6.4
Hz), 4.36 (s, H), 4.14 (d, H, J=12 Hz), 3.92 (d, H, J=12.4 Hz),
3.02 (br s, OH), 2.83 (m, H), 1.90 (d, 3H, J=1.2 Hz), 1.02 (d, 3H,
J=6.8 Hz), 0.94 (s, 9H), 0.17 (s, 3H), 0.14 (s, 3H).
Synthesis of Compound 70
##STR00117##
[0239] To a well-dried mixture of substrate 68 (35 mg, 0.09 mmol)
and ammonium fluoride (33 mg, 0.73 mmol) was added MeOH (3 mL) and
refluxed at 85.degree. C. The reaction completed in 12 h. The
solvent was removed and the crude product was dissolved in 20%
MeOH/CH.sub.2Cl.sub.2 solution. The reaction mixture was filtered
through a celite pad. The solvent was removed under reduced
pressure. The crude product was purified by silica gel
chromatography, eluting with 1-8% MeOH/CH.sub.2Cl.sub.2 to afford
product 70 as colorless solid (20 mg, 80%). Mass calcd: 281. 27.
Found: 282.20 (M.sup.++H); .sup.1H NMR (CDCl.sub.3) .delta. 7.73
(s, H), 6.34 (d, H, J=6.4 Hz), 4.19 (d, H, J=10.4 Hz), 4.02 (d, H,
J=12.4 Hz), 3.92 (d, H, J=12 Hz), 2.73 (m, H), 1.85 (s, 3H), 0.98
(d, 3H, J=6.8 Hz).
Preparation of Triethynyl Aluminum
[0240] To a solution of AlCl.sub.3 (1 g, 7.5 mmol) in
CH.sub.2Cl.sub.2 (7.5 mL) was added ethynyl magnesium chloride
(37.5 mL from 0.6 M, in THF, 22.5 mmol) under Ar at 0.degree. C.
The reaction mixture was warmed to RT and stirred for over night.
The resulting dark brown solution (0.14 M) was used for next
step.
Synthesis of Compounds 71 and 72
##STR00118##
[0242] To the cold solution of epoxide 64 (80 mg, 0.22 mmol) in
CH.sub.2Cl.sub.2 (6 mL) at -30.degree. C. was added triethynyl
aluminum (5.2 mL from 0.14 M stock solution in
CH.sub.2Cl.sub.2/THF, 0.56 mmol) at once under argon atmosphere and
stirred at -30.degree. C. for 6 h. The reaction was quenched with
saturated NH.sub.4Cl solution and filtered through a celite pad.
The filtrate was partitioned between CH.sub.2Cl.sub.2/H.sub.2O. The
organic layer was separated and dried (Na.sub.2SO.sub.4). The
solvent was removed and the crude product was purified by silica
gel chromatography, eluting with 5-30% ethyl acetate/hexane to
afford products 71 and 72 as semi solid (57 mg, over all 67%). Mass
calcd: 394.20. Found: 395.40 (M.sup.++H).
[0243] Isomer 71: .sup.1H NMR (CDCl.sub.3): .delta. 9.02 (s, NH),
7.58 (s, H), 6.31 (s H), 4.10 (d, H, J=12.0 Hz), 3.99 (d, H, J=12.6
Hz), 3.71 (d, H, J=12.3 Hz), 2.81 (m, H), 1.91 (s, 3H), 1.01 (s,
3H), 0.97 (s, 9H), 0.17 (s, 3H), 0.12 (s. 3H).
[0244] Isomer 72: .delta. 8.34 (s, NH), 7.43 (s, H), 6.33 (d, H,
J=7.2 Hz), 4.23 (d, H, J=7.2 Hz), 3.91 (dd, H, J=12.4 Hz, 4.8 Hz),
3.78 (dd, H, J=12.0 Hz, 9.2 Hz), 2.79 (m, H), 1.95 (s, 3H), 0.98
(d, H, J=7.6 Hz), 0.93 (s, 9H), 0.18 (s, 3H), 0.14 (s, 3H).
Synthesis of Compound 73
##STR00119##
[0246] To a well dried mixture of substrate 71 (20 mg, 0.46 mmol)
and ammonium fluoride (17 mg, 4.56 mmol) was added MeOH (3 mL) and
refluxed at 85.degree. C. Reaction completed in 12 h, the solvent
was removed by reduced pressure and the crude product was dissolved
in 20% MeOH/CH.sub.2Cl.sub.2 solution. The reaction mixture was
filtered through a celite pad.
[0247] The solvent was removed under reduced pressure and the crude
product was purified by silica gel chromatography, eluting with
1-8% MeOH/CH.sub.2Cl.sub.2 to afford product 73 as colorless solid
(12 mg, 94%). Mass calcd: 280.30. Found: 281.20 (M.sup.++H);
.sup.1H NMR (CDCl.sub.3) .delta. 7.94 (s, H), 6.23 (d, H, J=8 Hz),
4.03 (d, H, J=10.8 Hz), 3.90 (d, H, J=12.4 Hz), 3.80 (d, H, J=12.4
Hz), 3.05 (s, H), 2.75 (m, H), 1.85 (d, 3H, J=1.2 Hz), 0.95 (d, 3H,
J=7.2 Hz).
Synthesis of Compound 74
##STR00120##
[0249] To a well-dried mixture of substrate 72 (10 mg, 0.03 mmol)
and ammonium fluoride (10 mg, 0.3 mmol) was added MeOH (2 mL) and
refluxed at 85.degree. C. The reaction completed in 12 h and the
solvent was removed under reduced pressure. The crude product was
dissolved in 20% MeOH/CH.sub.2Cl.sub.2 solution. The reaction
mixture was filtered through a celite pad. The solvent was removed
and the crude product was purified by silica gel chromatography,
eluting with 1-8% MeOH/CH.sub.2Cl.sub.2 to afford product 74 as
colorless solid (8 mg, 91%). Mass calcd: 280.30. Found: 281.2
(M.sup.++H); .sup.1H NMR (CDCl.sub.3) .delta. 7.58 (s, H), 6.30 (d,
H, J=8.4 Hz), 4.16 (d, H, J=12.2 Hz), 3.89 (d, H, J=12.8 Hz), 3.71
(d, H, J=12.7 Hz), 3.28 (s, H), 2.85 (s, H), 1.90 (d, 3H, J=1.3
Hz), 0.96 (d, 3H, J=7.0).
[0250] In a similar manner, but using the appropriate sugar and
pyrimidine or purine bases, the following nucleosides of formula as
indicated can be prepared.
TABLE-US-00004 ##STR00121## R.sup.2 R.sup.3 R.sup.4 R.sup.5 X
R.sup.6 CH.sub.3 OH N.sub.3 NH.sub.2, NH--NH.sub.2, NH--OH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2 , CH.sub.2OH
CH.sub.3 OH C.ident.CH NH.sub.2, NH--NH.sub.2, NH--OH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2 , CH.sub.2OH
CH.sub.3 OH CH.dbd.CH.sub.2 NH--NH.sub.2 , NH--OH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH
CH.sub.3 OH CN NH.sub.2, NH--NH.sub.2, NH--OH, O, S H, CH.sub.3, F,
Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2, CH.dbd.CHBr,
CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH CH.sub.3 OH OMe
NH.sub.2, NH--NH.sub.2, NH--OH, O, S H, CH.sub.3, F, Cl, Br, I,
NO.sub.2, OH, SH, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl,
C.ident.CH, CN, NH.sub.2, CH.sub.2OH CH.sub.3 OH F NH.sub.2,
NH--NH.sub.2, NH--OH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, OH,
SH, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN,
NH.sub.2, CH.sub.2OH CH.sub.3 OH Me NH.sub.2, NH--NH.sub.2, NH--OH,
O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH
CH.dbd.CH.sub.2 OH C.ident.CH NH.sub.2, NH--NH.sub.2, NH--OH, O, S
H, CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH
CH.dbd.CH.sub.2 OH CN NH.sub.2, NH--NH.sub.2, NH--OH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH
CH.dbd.CH.sub.2 OH F NH.sub.2, NH--NH.sub.2, NH--OH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH
CH.dbd.CH.sub.2 OH Me NH.sub.2, NH--NH.sub.2, NH--OH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH
CH.dbd.CH.sub.2 OH N.sub.3 NH.sub.2, NH--NH.sub.2, NH--OH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, 2 OH, SH, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH
C.ident.CH OH C.ident.CH NH.sub.2, NH--NH.sub.2, NH--OH, O, S H,
CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH
C.ident.CH OH CN NH.sub.2, NH--NH.sub.2, NH--OH, O, S H, CH.sub.3,
F, Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2, CH.dbd.CHBr,
CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH C.ident.CH OH
N.sub.3 NH.sub.2, NH--NH.sub.2, NH--OH, O, S H, CH.sub.3, F, Cl,
Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl,
C.ident.CH, CN, NH.sub.2, CH.sub.2OH C.ident.CH OH OMe NH.sub.2,
NH--NH.sub.2, NH--OH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, OH,
SH, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN,
NH.sub.2, CH.sub.2OH CH.sub.2F OH C.ident.CH NH.sub.2,
NH--NH.sub.2, NH--OH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, OH,
SH, CH.dbd.CH.sub.2,CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN,
NH.sub.2, CH.sub.2OH CH.sub.2F OH CN NH.sub.2, NH--NH.sub.2,
NH--OH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH,
CH.dbd.CH.sub.2,CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2,
CH.sub.2OH CH.sub.2F OH N.sub.3 NH.sub.2, NH--NH.sub.2, NH--OH, O,
S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH,
CH.dbd.CH.sub.2,CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2,
CH.sub.2OH CH.sub.2F OH CH.dbd.CH.sub.2 NH.sub.2, NH--NH.sub.2,
NH--OH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH,
CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN,
NH.sub.2, CH.sub.2OH CN OH C.ident.CH NH.sub.2, NH--NH.sub.2,
NH--OH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH,
CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN,
NH.sub.2, CH.sub.2OH CN OH CN NH.sub.2, NH--NH.sub.2, NH--OH, O, S
H, CH.sub.3, F, Cl, Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2,
CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN, NH.sub.2, CH.sub.2OH CN
OH N.sub.3 NH.sub.2, NH--NH.sub.2, NH--OH, O, S H, CH.sub.3, F, Cl,
Br, I, NO.sub.2, OH, SH, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl,
C.ident.CH, CN, NH.sub.2, CH.sub.2OH CN OH CH.dbd.CH.sub.2
NH.sub.2, NH--NH.sub.2, NH--OH, O, S H, CH.sub.3, F, Cl, Br, I,
NO.sub.2, OH, SH, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl,
C.ident.CH, CN, NH.sub.2, CH.sub.2OH CH.sub.2CH.sub.3 OH N.sub.3
NH.sub.2, NH--NH.sub.2, NH--OH, O, S H, CH.sub.3, F, Cl, Br, I,
NO.sub.2, OH, SH, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl,
C.ident.CH, CN, NH.sub.2, CH.sub.2OH CF.sub.3 OH N.sub.3 NH.sub.2,
NH--NH.sub.2, NH--OH, O, S H, CH.sub.3, F, Cl, Br, I, NO.sub.2, OH,
SH, CH.dbd.CH.sub.2, CH.dbd.CHBr, CH.dbd.CHCl, C.ident.CH, CN,
NH.sub.2, CH.sub.2OH
TABLE-US-00005 ##STR00122## R.sup.2 R.sup.3 R.sup.4 R.sup.6 R.sup.5
Z R.sup.8 R.sup.7 CH.sub.3 OH N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH-
N -- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH-
cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2,N,N-Et.sub.2, F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH
N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH N.sub.3 NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.3 OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N, N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.sub.3 OH C.ident.CH H NH.sub.2, NH-Me, NH-Et, NH- N --
H propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH C.ident.CH H
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH C.ident.CH NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.3 OH C.ident.CH NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.sub.3 OH CH.dbd.CH.sub.2 H NH.sub.2, NH-Me, NH-Et, NH-
N -- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH-
cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH
CH.dbd.CH.sub.2 H NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H
propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl,
NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH,
CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br,
I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH
CH.dbd.CH.sub.2 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl,
heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH
CH.dbd.CH.sub.2 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl,
Br, I, H propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2,
cyclopropyl, NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl,
COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH CN
H NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.3 OH CN H NH.sub.2, NH-Me, NH-Et, NH- C
H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.sub.3 OH CN NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- N -- H
propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH CN NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH OMe H NH.sub.2, NH-Me,
NH-Et, NH- N -- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl,
NH- cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH
OMe H NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH
OMe NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl,
heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH OMe NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH F H NH.sub.2, NH-Me,
NH-Et, NH- N -- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl,
NH- cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH
F H NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH F NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH- cyclopropyl,
NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.sub.3 OH F NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- C H, F,
Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2,
cyclopropyl, NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl,
COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH Me
H NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.3 OH Me H NH.sub.2, NH-Me, NH-Et, NH- C
H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.sub.3 OH Me NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- N -- H
propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.3 OH Me NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH C.ident.CH H
NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH C.ident.CH H NH.sub.2, NH-Me,
NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.dbd.CH.sub.2 OH C.ident.CH NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- N -- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl,
NH- cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl
CH.dbd.CH.sub.2 OH C.ident.CH NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-
C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.dbd.CH.sub.2 OH CN H NH.sub.2, NH-Me, NH-Et, NH- N -- H
propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH CN H
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, COOMe Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH CN NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH CN NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.dbd.CH.sub.2 OH F H NH.sub.2, NH-Me, NH-Et, NH- N -- H
propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH F H
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH F NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH F NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.dbd.CH.sub.2 OH Me H NH.sub.2, NH-Me, NH-Et, NH- N -- H
propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH Me H
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH Me NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.dbd.CH.sub.2 OH Me NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.dbd.CH.sub.2 OH N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH- N
-- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH-
cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl
CH.dbd.CH.sub.2 OH N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH- C H, F,
Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2,
cyclopropyl, NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl,
COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl
CH.dbd.CH.sub.2 OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- N
-- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH-
cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl
CH.dbd.CH.sub.2 OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- C
H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl C.ident.CH OH C.ident.CH H NH.sub.2, NH-Me, NH-Et, NH- N
-- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH-
cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH
C.ident.CH H NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H
propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl,
NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH,
CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br,
I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH
C.ident.CH NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl,
heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH C.ident.CH
NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH CN H NH.sub.2,
NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH- cyclopropyl,
NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl C.ident.CH OH CN H NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl,
Br, I, H propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2,
cyclopropyl, NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl,
COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH
CN NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle,
NH- cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl C.ident.CH OH CN NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl C.ident.CH OH N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH- N -- H
propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH N.sub.3 H
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH N.sub.3 NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe,
SEt, O-acetyl C.ident.CH OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl C.ident.CH OH OMe H NH.sub.2, NH-Me, NH-Et, NH- N -- H
propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH OMe H
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl C.ident.CH OH OMe NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl C.ident.CH OH OMe NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.sub.2F OH C.ident.CH H NH.sub.2, NH-Me, NH-Et, NH- N --
H propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH C.ident.CH H
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH C.ident.CH NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.2F OH C.ident.CH NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH-
NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH,
CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn,
SH, SMe, SEt, O-acetyl CH.sub.2F OH CN H NH.sub.2, NH-Me, NH-Et,
NH- N -- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH-
cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH
CN H NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH CN NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH- cyclopropyl,
NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.sub.2F OH CN NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- C H,
F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2,
cyclopropyl, NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl,
COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH
N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle,
NH- cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.2F OH N.sub.3 H NH.sub.2, NH-Me, NH-Et,
NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CH.sub.2F OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH-
N -- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH-
cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH
N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H
propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl,
NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH,
CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br,
I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH
CH.dbd.CH.sub.2 H NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl,
heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH
CH.dbd.CH.sub.2 H NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H
propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl,
NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH,
CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br,
I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH
CH.dbd.CH.sub.2 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl,
heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2F OH
CH.dbd.CH.sub.2 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl,
Br, I, H propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2,
cyclopropyl, NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl,
COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH
C.ident.CH H NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl,
heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH C.ident.CH H
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CN OH C.ident.CH NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CN OH C.ident.CH NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH, CN,
cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CN OH CN H NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl,
heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH CN H NH.sub.2,
NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH-
NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH,
CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn,
SH, SMe, SEt, O-acetyl CN OH CN NH.sub.2 NH.sub.2, NH-Me, NH-Et,
NH- N -- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH-
cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH CN
NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CN OH N.sub.3 H NH.sub.2, NH-Me,
NH-Et, NH- N -- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl,
NH- cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH
N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CN OH N.sub.3 NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH- cyclopropyl,
NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2,
N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt,
O-acetyl CN OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- C H, F,
Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2,
cyclopropyl, NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl,
COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH
CH.dbd.CH.sub.2 H NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl,
heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CN OH CH.dbd.CH.sub.2 H
NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CN OH CH.dbd.CH.sub.2 NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CN OH NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- C H,
F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2,
cyclopropyl, NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl,
COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl
CH.sub.2CH.sub.3 OH N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH- N -- H
propyl, heterocycle, NH- cyclopropyl, NH-acetyl, NH- cyclobutyl,
NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I,
OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2CH.sub.3 OH
N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CH.sub.2CH.sub.3 OH N.sub.3
NH.sub.2 NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle,
NH- cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CH.sub.2CH.sub.3 OH N.sub.3 NH.sub.2 NH.sub.2,
NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH-
NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl, NH- C.ident.CH,
CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe, OEt, OBn,
SH, SMe, SEt, O-acetyl CF.sub.3 OH N.sub.3 H NH.sub.2, NH-Me,
NH-Et, NH- N -- H propyl, heterocycle, NH- cyclopropyl, NH-acetyl,
NH- cyclobutyl, NH-t-butyl, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2,
F, Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl CF.sub.3 OH
N.sub.3 H NH.sub.2, NH-Me, NH-Et, NH- C H, F, Cl, Br, I, H propyl,
heterocycle, NH- NO.sub.2, CH.dbd.CH.sub.2, cyclopropyl, NH-acetyl,
NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl, COOH, CONH.sub.2,
Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F, Cl, Br, I, OH, OMe,
OEt, OBn, SH, SMe, SEt, O-acetyl CF.sub.3 OH N.sub.3 NH.sub.2
NH.sub.2, NH-Me, NH-Et, NH- N -- H propyl, heterocycle, NH-
cyclopropyl, NH-acetyl, NH- cyclobutyl, NH-t-butyl, Azetidine,
N,N-Me.sub.2, N,N-Et.sub.2, F, Cl, Br, I, OH, OMe, OEt, OBn, SH,
SMe, SEt, O-acetyl CF.sub.3 OH N.sub.3 NH.sub.2 NH.sub.2, NH-Me,
NH-Et, NH- C H, F, Cl, Br, I, H propyl, heterocycle, NH- NO.sub.2,
CH.dbd.CH.sub.2,
cyclopropyl, NH-acetyl, NH- C.ident.CH, CN, cyclobutyl, NH-t-butyl,
COOH, CONH.sub.2, Azetidine, N,N-Me.sub.2, N,N-Et.sub.2, COOMe F,
Cl, Br, I, OH, OMe, OEt, OBn, SH, SMe, SEt, O-acetyl
Biological Assays
HBV AD38 Assay
[0251] The materials required for the assay comprise the
following:
[0252] The HepG2- AD38 cell line.
[0253] The culture medium for HepG2- AD38 comprises DMEM-F/12, 10%
fetal bovine serum, 100 IU/ml/100 ug/ml of Penicillin/streptomycin,
50 .mu.g/ml kanamycin, 0.3 .mu.g/ml tetracycline, and 200 g/ml
G418.
[0254] The assay medium for HepG2-AD38 comprises DMEM-F/12, 10%
fetal bovine serum, 100 IU/ml/100 .mu.g/ml of
penicillin/streptomycin, 50 .mu.g/ml kanamycin, and 200 .mu.g/ml
G418
[0255] Further materials comprise Phosphate buffered saline (PBS),
Biocoated 96 well plates, DNeasy 96 tissue kit (Qiagen), QIAvac 96
vacuum manifold, Micro amp optical 96 well reaction plates (Applied
Biosystems), Micro amp optical caps (Applied Biosystems), Tagman
Universal PCR Master Mix (Applied Biosystems), and a 7700 Sequence
detector (Applied Biosystems), The primers and probes for HBV DNA
comprise the 1125 nM forward primer (GGA CCC CTG CTC GTG TTA CA),
the 1125 nM reverse primer (GAG AGA AGT CCA CCA CGA GTC TAG A), and
the 250 nM probe (FAM-TGT TGA CAA GAA TCC TCA CAA TAC CAC).
HBV Cell Assay:
[0256] Seed 5.times.10.sup.4 cells/well in 200 .mu.l of culture
medium in a 96 well biocoated plate and incubate the plate at
37.degree. C. with 5% CO.sub.2. After 2 days, remove carefully the
supernatant, wash the cell layer with 200 .mu.l of PBS and renew
with 200 .mu.l of assay medium with or without test compounds at 10
.mu.M or in dose response with ratio 1:3 starting at 10 .mu.M (all
samples should be tested in duplicate). Let cells grow for 5 more
days. At day 7, collect 180 .mu.l of the supernatant per well in a
blue rack (included in the DNeasy 96 tissue kit). Store at
-80.degree. C. or directly go to next step.
Extraction of Viral HBV DNA from Cell Supernatant:
[0257] Thaw the samples in the blue rack. Prepare a Proteinase
K/Buffer ATL working solution (2 ml of Proteinase K+18 ml Buffer
ATL) and transfer 180 ul on the top of the supernatant in each tube
of the blue rack. Seal the tubes properly using the caps provided
and mix by inverting the rack upside down a few times. Centrifuge
up to 300 rpm to collect any solution from the caps. Incubate at
55.degree. C. for 15 minutes. Centrifuge up to 300 rpm again.
Carefully remove the caps and add 410 ul of Buffer AL/E to each
sample. Seal the tubes using new caps, shake the rack vigorously up
and down for 15 seconds and centrifuge up to 3000 rpm. Place the
DNeasy 96 plate on top of QIAvac 96 vacuum manifold. Transfer the
supernatant from step 8 to the DNeasy 96 plate. Apply the vacuum
for a few seconds. Carefully add 500 .mu.l of Buffer AW1 to each
well. Apply the vacuum for about 1 minute. Carefully add 500 .mu.l
of Buffer AW2 to each well. Apply the vacuum for about 1 minute,
flick the plate in the sink, strike the bottom side of the DNeasy
96 plate on a stack of paper towels, apply the vacuum again for 10
minutes. Heat up 10 ml of Buffer AE for a few minutes at 70.degree.
C. Place the DNeasy 96 plate on top of a rack of elution microtubes
RS. To elute the DNA, add 100 .mu.l of preheated Buffer AE to each
well, apply the vacuum for 1 minute.
HBV Real Time PCR:
[0258] Prepare the HBV primers+probe mix for 200 wells (total 1500
.mu.l) comprising 45 .mu.l of primer 1 (100 .mu.M), 45 .mu.l of
primer 2 (100 .mu.M), 20 .mu.l of probe (50 .mu.M), and 1390 .mu.l
of nuclease free water. Select an optical 96 well reaction plate.
Make the reaction mix for 100 wells comprising 1000 .mu.l of
Universal PCR Master Mix, 750 ul of HBV primers+probe mix, and 250
.mu.l of nuclease free water. Aliquot 20 .mu.l of the reaction mix
per well. Add 5 .mu.l per well of HBV DNA from each sample. Cover
the wells with optical caps. Centrifuge for a few seconds to force
all the reagents to the bottom and to eliminate the bubbles. Place
the plate in the 7700 Sequence detector. Select the reporter for
FAM and the volume setting for 25 .mu.l. Start the machine and let
it go 1 hour and 56 minutes. Calculate the dCt and the reduction in
viral load for each test compound.
HCV Replicon Assay
[0259] HCV replicon RNA-containing Huh 7 cells (clone A cells;
Apath, LLC, St. Louis, Mo.) were kept at exponential growth in
Dulbecco's modified Eagle's medium (high glucose) containing 10%
fetal bovine serum, 4 mM L-glutamine and 1 mM sodium pyruvate,
1.times. nonessential amino acids, and G418 (1,000 .mu.g/ml).
Antiviral assays were performed in the same medium without G418.
Cells were seeded in a 96-well plate at 1,500 cells per well, and
test compounds were added immediately after seeding. Incubation
time 4 days. At the end of the incubation step, total cellular RNA
was isolated (RNeasy 96 kit; Qiagen). Replicon RNA and an internal
control (TaqMan rRNA control reagents; Applied Biosystems) were
amplified in a single-step multiplex RT-PCR protocol as recommended
by the manufacturer. The HCV primers and probe were designed with
Primer Express software (Applied Biosystems) and covered highly
conserved 5'-untranslated region (UTR) sequences (sense,
5'-AGCCATGGCGTTAGTA(T)GAGTGT-3', and antisense,
5'-TTCCGCAGACCACTATGG-3'; probe,
5'-FAM-CCTCCAGGACCCCCCCTCCC-TAMRA-3').
[0260] To express the antiviral effectiveness of a compound, the
threshold RT-PCR cycle of the test compound was subtracted from the
average threshold RT-PCR cycle of the no-drug control
(.DELTA.CtHCV). A .DELTA.Ct of 3.3 equals a 1-log 10 reduction
(equal to the 90% effective concentration [EC.sub.90]) in replicon
RNA levels. The cytotoxicity of the test compound could also be
expressed by calculating the .DELTA.CtrRNA values. The
.DELTA..DELTA.Ct specificity parameter could then be introduced
(.DELTA.CtHCV-.DELTA.CtrRNA), in which the levels of HCV RNA are
normalized for the rRNA levels and calibrated against the no-drug
control.
HIV Activity
[0261] HIV screen: Primary Screening of PSI compounds are tested
for antiviral HIV activity at 50M. The cells used are P4CCR5luc
cells; they are human HIV indicator cells, which are derived from
Hela cells, express CD4, CXCR4, CCR5, luciferase, and a beta-gal
gene under the control of HIV-1 LTR. P4CCR5 luc cells are
cultivated in DMEM, 10% FBS, Penicillin, Streptomycin, and G418 at
500 .mu.g/ml. 100 .mu.l of P4 CCR5-luc cells are plated at 10,000
cells per well in 96 well Opaque Assay plates and incubated
overnight at 37.degree. C. The next day, the media is aspirated
from the plates and replaced by 100 .mu.l of compound freshly
diluted into media at 2.times.50 uM, in triplicate, for 4 hours at
37.degree. C. The cells are then infected with 100 ul NL43 virus at
5 ng of p24 per well, in the presence of 2.times.20 ug/ml of
DEAE-Dextran for 40-42 hours. Non infected, infected no drug and
AZT controls are always present in triplicate on each plate. After
infection the beta-gal is quantitated using the Galacto-Star kit
from Applied Biosystems using the manufacturer instructions and the
luminescence measured using a Victor apparatus from Perkin-Elmer.
Results are represented as percentage inhibition compare to
untreated cells. The assays are performed in 2 to 3 independent
experiments.
HIV-Titration of PSI Activity to Determine EC.sub.50 on P4 CCR-Luc
Cells.
[0262] P4 CCR5-luc cells are plated at 10,000 cells per well (100
.mu.l) in 96 well Opaque Assay plates and incubated overnight at
37.degree. C. The next day, the media is aspirated from the plates
and replaced by 100 ul of compound freshly diluted into appropriate
media (DMEM, 10% FBS, G418 500 .mu.g/ml, penicillin/streptomycin)
at 2.times. final concentrations in 5 fold dilutions, usually from
2.times.100 .mu.M to 2.times.0.032 uM, in triplicate, for 4 hours
at 37.degree. C. The cells are then infected with 100 ul NL43 wild
type or mutant virus, at 5 ng to 20 ng of p24 per well, in the
presence of 2.times.20 ug/ml of DEAE-Dextran, for 40-42 hours. Non
infected and infected no drug controls are always present in
12plicate on each plate. An AZT control is tested in parallel for
each experiment. After infection, the beta-gal is quantitated in
the cell lysate using the Galacto-Star kit from Applied Biosystems
and the luminescence measured using a Victor apparatus from
Perkin-Elmer. The EC.sub.50 (Effective Concentration) is calculated
using a Microsoft.RTM. Excel.RTM. spreadsheet that calculates the
concentration necessary to inhibit the 50% of the infection. The
assay is performed in at least 2 independent experiments.
Toxicity
Luciferase Assay
[0263] P4 CCR5-luc cells are plated at 10,000 cells per well (100
ul) in 96 well Opaque Assay plates and incubated overnight at
37.degree. C. The next day, the media is aspirated from the plates
and replaced by 200 ul of compound freshly diluted into media in 5
fold dilutions from 100 .mu.M to 0.0062 .mu.M. After 4 days of
incubation at 37.degree. C., the luciferase activity is measured in
the cell lysate using the Bright Glow kit from Promega and the
luminescence measured using a Victor apparatus from
Perkin-Elmer.
MTS Assays
[0264] Human cells lines Huh 7 and HepG2 (liver), BxPC3
(pancreatic) and CEM (lymphoid) are used for the MTS assays in 96
wells plates. Drugs are freshly diluted in media at 2.times.100
.mu.M, 50 .mu.M, 25 .mu.M, 10 .mu.M, 5 .mu.M, 1 .mu.M and 50 .mu.l
is dispensed in triplicate in the plates. The wells at the
periphery of the plate contain 100 ul of media only and will be the
blank controls. A replicate control with no drug is always
performed in each plate. 50 .mu.l of cells are added to the plate,
at 2000 cells per well for Huh 7, HepG2 and PxPC3, and 5000 cells
per well for CEM cells. No cells are added at the periphery of the
plate. The media used for Huh 7, HepG2 and BxPC3 cells is DMEM with
10% FBS, and Penicillin/streptomycin, and RPMI with 10% FBS, and
Penicillin/streptomycin for CEM cells. After 8 days of incubation
at 37.degree. C., 20 .mu.l of MTS dye from the CellTiter 96 Aqueous
One Solution Cell Proliferation Assay kit from Promega is added to
each well and the plate incubated for 2 h at 37.degree. C. The
absorbance is then read at 490 nm using the microplate reader E1800
from Biotek. The signal is calculated by subtracting the absorbance
measured in the blank controls. The CC.sub.50 (Cytotoxic
Concentration) value is then determined by comparing the signal
obtained with the no-drug cell control with the treated cells and
calculating the concentration of drug necessary to inhibit 50% of
the signal in the wells treated with drugs.
Biological Results
##STR00123##
[0265] Antiviral Activity
TABLE-US-00006 [0266] Screen HIV P4 Cells % Inhibition at 50 .mu.M
100.11 HIV Inhibition P4 Cells EC.sub.50 (.mu.M) 0.18 Screen HCV
Clone A Cells -log.sub.10 at 50 .mu.M (HCV:Rbz) {-1.4:-0.59} HCV
Inhibition Clone A cells EC.sub.90 (.mu.M) 7.85 HCV Inhibition
S282T Cells EC.sub.90 (.mu.M) 73.08 HBV Inhibition EC.sub.90
(.mu.M) 1.62
Cytotoxicity (CC.sub.50 .mu.M)
TABLE-US-00007 [0267] P4 Cells >100 CEM 400 BxPC3 400 HepG2 300
PBMC >100 Huh 7 >100
[0268] The present application claims priority to U.S. provisional
patent application 60/989,296, filed Nov. 20, 2007, the contents of
which are incorporated by reference in its entirety.
Sequence CWU 1
1
6120DNAArtificial SequenceHBV DNA FORWARD PRIMER 1ggacccctgc
tcgtgttaca 20225DNAArtificial SequenceHBV DNA REVERSE PRIMER
2gagagaagtc caccacgagt ctaga 25327DNAArtificial SequenceHBV DNA
PROBE WITH 5'-FAM LABEL 3tgttgacaag aatcctcaca ataccac
27423DNAArtificial SequenceHCV SENSE PRIMER 4agccatggcg ttagtatgag
tgt 23518DNAArtificial SequenceHCV ANTISENSE PRIMER 5ttccgcagac
cactatgg 18620DNAArtificial SequenceHCV PROBE WITH 5'-FAM LABEL AND
3'-TAMRA LABEL 6cctccaggac cccccctccc 20
* * * * *